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Dietary factors that influence amino acid, protein and nucleic acid metabolism in the chicken Akinwande, Amos Ifeolu 1973

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DIETARY FACTORS THAT INFLUENCE AMINO ACID, PROTEIN AND NUCLEIC ACID METABOLISM IN THE CHICKEN B Y AMOS IFEOLU AKINWANDE B.Sc . A G R I C . ( H O N S ) , U N I V E R S I T Y O F M A N I T O B A , 1969 M . S c . , U N I V E R S I T Y O F M A N I T O B A , 1970 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY I N T H E D E P A R T M E N T OF. P O U L T R Y S C I E N C E WE A C C E P T T H I S T H E S I S A S C O N F O R M I N G TO T H E R E Q U I R E D S T A N D A R D THE UNIVERSITY OF BRITISH COLUMBIA A P R I L , 1973 In presenting this thesis in partial fulfilment of the requirements for an advanced degree at the University of British Columbia, I agree that the Library shall make i t freely available for reference and study. I further agree that permission for extensive copying of this thesis for scholarly purposes may be granted by the Head of my Department or by his representatives. It is understood that copying or publication of this thesis for financial gain shall not be allowed without my written permission. Department The University of British Columbia Vancouver 8, Canada Date M,Oaj I . H73 ABSTRACT THE M E T A B O L I S M O F DNA, RNA, P R O T E I N A N D A M I N O A C I D S W/A"S:E S T U D I E D I N C H I C K S F E D D I E T A R Y P R O T E I N S O F D I F F E R E N T N U T R I T I V E V A L U E . DNA A N D RNA C O N T E N T O F T H E G A S T R O C N E M I U S M U S C L E A N D L I V E R W E R E M E A S U R E D B Y M E A N S O F A N U L T R A V I O L E T A B S O R P T I O N T E C H N I Q U E A N D P R O T E I N S G R A V I M E T R I C A L L Y A N D B Y S P E C T R O P H O T O M E T R Y . A N I S O T O P E T E C H N I Q U E WAS U S E D TO S T U D Y T H E E F F E C T S O F D I E T A R Y L Y S I N E ON T H E D E G R A D A T I O N A N D S Y N T H E S I S O F T I S S U E P R O T E I N S , RNA S Y N T H E S I S A N D T H E M E T A B O L I C F A T E O F L Y S I N E A N D T H R E O N I N E . R E S U L T S S H O W E D T H A T M U S C L E DNA (MG./G. F R E S H T I S S U E ) C O N T E N T WAS U N A F F E C T E D B Y T H E L E V E L O F D I E T A R Y L Y S I N E A T 14 A N D 28 D A Y S O F A G E . L I V E R DNA C O N T E N T I N C R E A S E D A T 14 D A Y S A N D WAS U N A F F E C T E D A T 28 D A Y S W I T H I N C R E A S E D L E V E L O F D I E T A R Y L Y S I N E . M U S C L E P R O T E I N / D N A A N D RNA/DNA R A T I O S I N C R E A S E D A T 14 A N D 28 D A Y S O F A G E B Y I N C R E A S I N G D I E T A R Y P R O T E I N F R O M 15 TO 20$, B Y S U P P L E M E N T I N G W H E A T W I T H S O Y B E A N M E A L OR W I T H L Y S I N E . T H E R A T I O S D E C R E A S E D A T D I E T A R Y L Y S I N E L E V E L S H I G H E R T H A N 0.86$. L I V E R P R O T E I N / D N A A N D RNA/DNA D E C R E A S E D P R O G R E S S I V E L Y W I T H I N C R E A S E D D I E T A R Y P R O T E I N A N D L Y S I N E A T 14 D A Y S O F A G E A N D I N C R E A S E D A T 28 D A Y S . L I V E R P R O T E I N / R N A R A T I O A T B O T H 14 A N D 28 D A Y S WAS C O N S T A N T A N D I N D E P E N D E N T O F L Y S I N E I N T A K E I N D I C A T I N G T H A T RNA I S C O R R E L A T E D W I T H P R O T E I N B I O S Y N T H E S I S . P R O T E I N A C C U M U L A T I O N WAS D E P R E S S E D MORE S E V E R E L Y I N B O T H I T I S S U E S T H A N DNA B Y P R O T E I N A N D L Y S I N E D E F I C I E N C Y . E X C E S S D I E T A R Y L Y S I N E D E P R E S S E D F E E D I N T A K E , C H I C K G R O W T H A N D P R O T E I N / D N A A N D RNA/DNA I N B O T H T I S S U E S . T H E S E D E P R E S S I O N S W E R E A L L E V I A T E D B Y S U P P L E M E N T A R Y A R G I N I N E . • H A L F - L I F E O F 7.2, 4.9 A N D 5.2 D A Y S C O R R E S P O N D I N G TO 10.4, 7.1 A N D 7.5 D A Y S T U R N O V E R T I M E , A N D 9.6, 14.0 A N D 13.3$ D A I L Y N E T D E G R A D A T I O N W E R E O B S E R V E D F O R L I V E R P R O T E I N S A T 0.36, 0.86 A N D 1.24$ D I E T A R Y L Y S I N E R E S P E C T I V E L Y . L I V E R R A T E O F P R O T E I N S Y N T H E S I S WAS 20.0, 49.2 A N D 52.9 M G . / D A Y R E S P E C T I V E L Y . THE T U R N O V E R R A T E S O F B O T H T H E M U S C L E A N D L I V E R P R O T E I N S I N C R E A S E D I N A M A N N E R S I M I L A R TO T H E G A I N I N B O D Y W E I G H T W I T H T H E L E V E L S O F D I E T A R Y L Y S I N E . L Y S I N E D E F I C I E N C Y D E P R E S S E D RNA S Y N T H E S I S I N T H E L I V E R M A R K E D L Y A N D I N T H E M U S C L E S L I G H T L Y . 14 L Y S I N E - U - C O X I D A T I O N I N C R E A S E D W H I L E R E T E N T I O N I N L I V E R A N D M U S C L E P R O T E I N S D E C L I N E D W I T H I N C R E A S I N G D I E T A R Y L Y S I N E . T H R E O N I N E -U - ^ C O X I D A T I O N WAS I N C R E A S E D A N D R E T E N T I O N D E C R E A S E D I N L Y S I N E D E F I C I E N C Y A N D T O X I C I T Y . THE F O L L O W I N G C O N C L U S I O N S A R E E V I D E N T F R O M R E S U L T S O F T H I S S T U D Y : (1) M U S C L E G R O W T H I N T H E C H I C K I S B Y A N I N C R E A S E I N C E L L S I Z E A N D NOT I N T H E N U M B E R O F C E L L S F R O M H A T C H TO 28 D A Y S O F A G E W H E R E A S T H E L I V E R U N D E R G O E S R A P I D C E L L M U L T I P L I C A T I O N D U R I N G T H E F I R S T 14 D A Y S A N D L I V E R G R O W T H A T 28 D A Y S I S M O S T L Y B Y C E L L H Y P E R T O P H Y . (2) P R O T E I N D E F I C I E N C Y I N H I B I T S C H I C K G R O W T H S E V E R E L Y A N D C A U S E S A R E D U C T I O N I N C E L L S I Z E W I T H NO D E G E N E R A T I O N O F C E L L S . THE R E S U L T E X P L A I N S WHY C H I C K S U N D E R S E V E R E P R O T E I N D E F I C I E N C Y A C H I E V E A NORMAL S I Z E WHEN R E H A B I L I T A T E D ON ADEQUATE P R O T E I N D I E T S . (3) A R G I N I N E BECOMES L I M I T I N G FOR T I S S U E GROWTH WHEN E X C E S S L Y S I N E I S P R E S E N T I N THE C H I C K D I E T . (4) TURNOVER RATE OF L I V E R P R O T E I N AND S Y N T H E S I S OF RNA I N L Y S I N E D E F I C I E N C Y ARE C O N S I D E R A B L Y LOWER THAN NORMAL. (5) A L A R G E P R O P O R T I O N OF L Y S I N E I S USED FOR P R O T E I N S Y N T H E S I S WHEN P R E S E N T I N S U B O P T I M A L AMOUNTS. I N C R E A S E D O X I D A T I O N OF THE C A R B O N S K E L E T O N TO CO^, OCCURS AS THE D I E T A R Y S U P P L Y E X C E E D S THE A N I M A L NEEDS FOR P R O T E I N S Y N T H E S I S . L I M I T E D P R O T E I N S Y N T H E S I S C A U S E D BY A L I M I T A T I O N I N D I E T A R Y L Y S I N E R E S U L T S I N A L O S S OF T H R E O N I N E WHEREAS WITH ADEQUATE L Y S I N E , P R O T E I N S Y N T H E S I S U T I L I Z E S A L A R G E P R O P O R T I O N OF D I E T A R Y L Y S I N E AND T H R E O N I N E , T H E R E B Y R E D U C I N G THE AMOUNTS OF T H R E O N I N E A V A I L A B L E FOR O X I D A T I O N . (6) C E L L U L A R DNA, RNA AND P R O T E I N C O N T E N T ARE U S E F U L P A R A M E T E R S FOR E V A L U A T I N G D I E T A R Y P R O T E I N S AND THE N U T R I T I O N A L S T A T U S OF THE AN I M A L . I I I TABLE OF CONTENTS PAGE ABSTRACT i LIST OF TABLES v I i LIST OF FIGURES i x LIST OF APPENDIX TABLES x i LIST OF APPENDIX FIGURES xv i ACKNOWLEDGEMENTS xv i I INTRODUCTION 1 LITERATURE REVIEW 4 C E L L U L A R B A S I S FOR CHANGES I N BODY C O M P O S I T I O N . . . 4 IMPACT OF D I E T A R Y P R O T E I N S ON T I S S U E P R O T E I N AND N U C L E I C A C I D S 7 N U T R I T I O N A L R E G U L A T I O N OF T I S S U E P R O T E I N AND RNA M E T A B O L I S M 12 T I S S U E P R O T E I N S Y N T H E S I S AND D E G R A D A T I O N IN R E L A T I O N TO D I E T A R Y P R O T E I N 17 T I S S U E RNA S Y N T H E S I S AND D E G R A D A T I O N I N R E L A T I O N TO D I E T A R Y P R O T E I N 22 M E T A B O L I S M OF S P E C I F I C AMINO A C I D S AS I N F L U E N C E D BY D I E T A R Y AMINO ACID CONTENT 27 CONTROL MECHANISM OF FEED I N T A K E 30 AMINO ACID I N T E R A C T I O N 36 EXPERIMENTAL PROCEDURE 39 GENERAL 39 T E C H N I Q U E S : Q U A N T I T A T I V E S E P A R A T I O N AND DETERMINA-T I O N OF DNA, RNA AND P R O T E I N 39 METHODS FOR S E Q U E N T I A L S E P A R A T I O N OF RNA, DNA, L I P I D AND P R O T E I N OF T I S S U E HOMOGENATES . . 39 D E T E R M I N A T I O N OF RNA, DNA AND P R O T E I N . . . . 40 P R O T E I N MEASUREMENT WITH F O L I N - P H E N O L REAGENT 42 i v PAGE T R I A L 1. DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N T I S S U E 43 T R I A L 2. DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N T I S S U E 43 T R I A L 3. DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N T I S S U E 45 T R I A L 4. TURNOVER RATE OF T I S S U E P R O T E I N 46 MEASUREMENTS OF R A D I O A C T I V I T Y I N T I S S U E P R O T E I N . . . 48 T R I A L 5. S Y N T H E S I S OF T I S S U E RNA . 48 MEASUREMENTS OF R A D I O A C T I V I T Y I N JTMSSUE RNA 49 T R I A L 6. M E T A B O L I C FATE OF AMINO A C I D S 50 T R I A L 7. M E T A B O L I C FATE OF AMINO A C I D S 51 MEASUREMENTS OF R A D I O A C T I V I T Y I N T I S S U E P R O T E I N S . . . 52 T R I A L 8. M E T A B O L I C FATE OF AMINO A C I D S 52 RESULTS AND DISCUSSION 55 T R I A L 1 55 SUMMARY AND C O N C L U S I O N . . . 78 T R I A L 2 80 SUMMARY AND C O N C L U S I O N 91 T R I A L 3 93 SUMMARY AND C O N C L U S I O N 109 T R I A L 4 111 SUMMARY AND C O N C L U S I O N 131 T R I A L 5 132 SUMMARY AND C O N C L U S I O N 137 , v PAGE T R I A L 6 139 SUMMARY AND C O N C L U S I O N 142 T R I A L 7 143 SUMMARY AND C O N C L U S I O N 148 T R I A L 8 149 SUMMARY AND C O N C L U S I O N 157 GENERAL SUMMARY AND CONCLUSION . . . . . . . . 159 BIBLIOGRAPHY 162 APPENDIX 181 LIST OF TABLES TABLE PAGE 1. C O M P O S I T I O N OF WHEAT BASAL DIET 44 2. C O M P O S I T I O N OF E X P E R I M E N T A L D I E T S (TRIAL 3) 47 3. G A S T R O C N E M I U S MUSCLE WEIGHT, DNA, RNA AND P R O T E I N CONTENT FROM C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR TWO WEEKS 56 4. L I V E R WEIGHT AND DNA CONTENT FROM C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR TWO WEEKS . 57 5. L I V E R RNA AND P R O T E I N CONTENT FROM C H I C K S FED GRAOED L E V E L S OF D I E T A R Y L Y S I N E FOR TWO WEEKS 58 6. BODY WEIGHT G A I N AND FEED INTAKE OF C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E 60 7. G A S T R O C N E M I U S MUSCLE WEIGHT, DNA, RNA.AND P R O T E I N CONTENT FROM C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR FOUR WEEKS 71 8. L I V E R WEIGHT AND DNA CONTENT FROM C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR FOUR WEEKS 72 9. L I V E R RNA AND P R O T E I N CONTENT FROM C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR FOUR WEEKS 73 10. G A S T R O C N E M I U S MUSCLE DNA, RNA AND P R O T E I N CONTENT OF C H I C K S FED NORMAL AND E X C E S S L Y S I N E FOR TWO WEEKS AND A R G I N I N E S U P P L E M E N T E D TO THE E X C E S S L Y S I N E D I E T FROM TWO TO FOUR WEEKS 81 11. L I V E R DNA, RNA AND P R O T E I N CONTENT OF C H I C K S FED NORMAL AND E X C E S S L Y S I N E FOR TWO WEEKS AND A R G I N I N E S U P P L E -MENTED TO THE E X C E S S L Y S I N E DIET FROM TWO TO FOUR WEEKS 82 12. DNA, RNA AND P R O T E I N CONTENT I N THE G A S T R O C N E M I U S MUSCLE OF C H I C K S FED D I F F E R E N T L E V E L S AND Q U A L I T Y OF D I E T A R Y P R O T E I N FOR TWO WEEKS 94 13. DNA, RNA AND P R O T E I N CONTENT I N THE G A S T R O C N E M I U S MUSCLE OF C H I C K S FED D I F F E R E N T L E V E L S AND Q U A L I T Y OF D I E T A R Y P R O T E I N FOR FOUR WEEKS 95 v I I TABLE PAGE 14. BODY WEIGHT G A I N , FEED I N T A K E , L I V E R DNA, RNA AND P R O T E I N CONTENT I N C H I C K S FED D I F F E R E N T . L E V E L S AND Q U A L I T Y OF D I E T A R Y P R O T E I N FOR TWO WEEKS 96 15. BODY WEIGHT G A I N , FEED I N T A K E , L I V E R DNA, RNA AND P R O T E I N I N C H I C K S FED D I F F E R E N T L E V E L S AND Q U A L I T Y OF D I E T A R Y P R O T E I N FOR FOUR WEEKS 97 16. CHANGES I N BODY AND L I V E R WEIGHTS OF C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E AND I N J E C T E D WITH L Y S I N E - U -14C 112 17. CHANGES I N P R O T E I N S AND RNA C O N C E N T R A T I O N I N THE MIXED T I B I A MUSCLES AND L I V E R OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - ^ C 113 18. E F F E C T OF D I E T A R Y L Y S I N E ON T I S S U E P R O T E I N D E G R A D A T I O N . 118 19. E F F E C T OF D I E T A R Y L Y S I N E L E V E L ON I N C O R P O R A T I O N OF L Y S I N E - U - ^ 4 C INTO THE T I S S U E S OF C H I C K S 130 20. E F F E C T S OF D I E T A R Y L Y S I N E ON L I V E R RNA S Y N T H E S I S . . . . 133 21. E F F E C T OF D I E T A R Y L Y S I N E ON MUSCLE RNA S Y N T H E S I S . . . . 134 22. E F F E C T OF D I E T A R Y L Y S I N E ON BODY WEIGHT, L I V E R WEIGHT AND AMOUNTS OF RNA I N C H I C K L I V E R AND MUSCLE 136 23. P E R C E N T A G E RECOVERY OF R A D I O A C T I V I T Y I N R E S P I R A T O R Y CO2 A F T E R I N J E C T I O N OF L Y S I N E - U - ^ C , BODY WEIGHT AND FEED I N T A K E OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND WITH THREONINE AND A R G I N I N E " . 140 24. P E R C E N T A G E RECOVERY OF R A D I O A C T I V I T Y I N C H I C K L I V E R AND MUSCLE P R O T E I N AFTER I N J E C T I O N OF L Y S I N E - U - 1 4 C 144 25. P E R C E N T A G E RECOVERY OF R A D I O A C T I V I T Y I N C H I C K L I V E R AND MUSCLE P R O T E I N AFTER I N J E C T I O N OF T H R E O N I N E - U - ^ C . . . 145 26. P E R C E N T A G E RECOVERY OF R A D I O A C T I V I T Y I N R E S P I R A T O R Y C 0 2 , L I V E R AND MUSCLE P R O T E I N AFTER I N J E C T I O N OF L Y S I N E - U -1 4C ( T R I A L 8) 150 27. P E R C E N T A G E RECOVERY OF R A D I O A C T I V I T Y I N R E S P I R A T O R Y CO2, L I V E R AND MUSCLE P R O T E I N A F T E R I N J E C T I O N OF T H R E O N I N E -U- 1 4C ( T R I A L 8) 153 28. E F F E C T OF D I E T A R Y L Y S I N E L E V E L ON THREONINE U T I L I Z A T I O N BY THE C H I C K 154 V I I I LIST OF FIGURES F I G U R E PAGE 1. FEED INTAKE P A T T E R N OF C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR TWO WEEKS . 61 2. AVERAGE D A I L Y G A I N S AND FE'ED E F F I C I E N C I E S OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR TWO WEEKS 63 3. G A S T R O C N E M I U S MUSCLE PROTEIN/DNA AND RNA/DNA RATIOS I N C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR TWO WEEKS 65 4. L I V E R PROTEIN/DNA, RNA/DNA AND PROTEIN/RNA RATIOS FROM C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR TWO WEEKS ' 69 5. G A S T R O C N E M I U S MUSCLE PROTEIN/DNA, RNA/DNA AND PROTEIN/RNA RATIOS FROM C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR FOUR WEEKS 75 6. L I V E R PROTEIN/DNA, RNA/DNA AND PROTEIN/RNA RATIOS FROM C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E FOR FOUR WEEKS 77 7. G A S T R O C N E M I U S MUSCLE PROTEIN/DNA AND RNA/DNA RATIOS AS A F F E C T E D BY E X C E S S D I E T A R Y L Y S I N E I N THE C H I C K 83 8. L I V E R PROTEIN/DNA AND RNA/DNA RATIOS AS A F F E C T E D BY E X C E S S D I E T A R Y L Y S I N E I N THE CH I C K 85 9. IMPROVEMENT I N MUSCLE PROTEIN/DNA AND RNA/DNA R A T I O S FROM C H I C K S FED E X C E S S L Y S I N E WITH A R G I N I N E 87 10. CHANGES I N L I V E R PROTEIN/DNA AND RNA/DNA R A T I O S FROM C H I C K S FED E X C E S S L Y S I N E WITH A R G I N I N E 88 11. E F F E C T S OF S U P P L E M E N T I N G HIGH L Y S I N E D I ETS WITH A R G I N I N E ON A C C U M U L A T I O N OF G A S T R O C N E M I U S MUSCLE DNA, RNA AND P R O T E I N 89 12. E F F E C T S OF S U P P L E M E N T I N G HIGH L Y S I N E D I E T S WITH A R G I N I N E ON A C C U M U L A T I O N OF L I V E R DNA, RNA, P R O T E I N AND BODY WEIGHT G A I N / C H I C K 90 V i XIX F I G U R E PAGE 13. E F F E C T S OF THE Q U A L I T Y AND Q U A N T I T Y OF D I E T A R Y P R O T E I N ON G A S T R O C N E M I U S MUSCLE PROTEIN/DNA AND RNA/DNA R A T I O S I N THE C H I C K AT TWO WEEKS OF AGE 100 14. E F F E C T S OF THE Q U A L I T Y AND QUANTITY OF D I E T A R Y P R O T E I N ON G A S T R O C N E M I U S MUSCLE PROTEIN/DNA AND RNA/DNA R A T I O S I N THE C H I C K AT FOUR WEEKS OF AGE 101 15. E F F E C T S OF THE Q U A L I T Y AND QUANTITY OF D I E T A R Y P R O T E I N ON L I V E R PROTEIN/DNA AND RNA/DNA RATIOS I N THE C H I C K AT Two WEEKS OF AGE 103 16. E F F E C T S OF THE Q U A L I T Y AND Q U A N T I T Y OF D I E T A R Y P R O T E I N ON L I V E R PROTEIN/DNA AND RNA/DNA R A T I O S I N THE C H I C K AT FOUR WEEKS OF AGE 104 17. E F F E C T S OF D I E T A R Y P R O T E I N L E V E L AND Q U A L I T Y ON R E L A T I V E A C C U M U L A T I O N OF DNA, RNA AND P R O T E I N I N THE C H I C K G A S T R O C N E M I U S MUSCLE BETWEEN 14 AND 28 DAYS OF AGE .' 106 18. E F F E C T S OF D I E T A R Y P R O T E I N L E V E L AND Q U A L I T Y ON R E L A T I V E A C C U M U L A T I O N OF DNA, RNA AND P R O T E I N I N C H I C K L I V E R AND BODY WEIGHT G A I N / C H I C K BETWEEN 14 AND 28 DAYS OF AGE 107 19. MUSCLE P R O T E I N , RNA AND PROTEIN/RNA OF C H I C K S FED D I F F E R N T L E V E L S OF D I E T A R Y L Y S I N E AND I N J E C T E D WITH L Y S I N E - C 1 4 ( U ) 116 20. L I V E R P R O T E I N , RNA AND PROTEIN/RNA RATIO I N C H I C K S FED D I F F E R E N T L E V E L S OF D I E T A R Y L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - 1 ^ C . . . * . . 117 14 21. RATE OF LOSS OF C-PROITEIN FROM L I V E R S OF C H I C K S R E C E I V I N G D I F F E R E N T L E V E L S OF D I E T A R Y L Y S I N E 121 14 22. RATE OF LOSS OF C - P R O T E I N FROM THE T I B I A MUSCLE OF C H I C K S FED D I F F E R E N T L E V E L S OF D I E T A R Y L Y S I N E . . . . . 124 23. CHANGES I N THE P E R C E N T A G E DECREASE OF S P E C I F I C A C T I V I T I E S OF THE L I V E R AND MIXED T I B I A MUSCLE T I S S U E P R O T E I N S OF C H I C K S FED GRADED L E V E L S OF D I E T A R Y L Y S I N E . 125 24. BODY WEIGHT G A I N PER C H I C K PER DAY OF F E E D I N G GRADED L E V E L S OF D I E T A R Y L Y S I N E 128 '. x LIST OF APPENDIX TABLES TABLE PAGE I. A N A L Y S I S OF V A R I A N C E OF AVERAGE BODY WEIGHT G A I N , FEED I N T A K E , G A S T R O C N E M I U S MUSCLE WEIGHT, L I V E R WEIGHT AND L I V E R WEIGHT PER 100G. BODY WEIGHT OF C H I C K S FED GRADED L E V E L S , OF L Y S I N E FOR Two WEEKS ( T R I A L 1) . 182 I I . A N A L Y S I S OF V A R I A N C E OF DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N ONE GRAM OF G A S T R O C N E M I U S MUSCLE OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR Two WEEKS (TRI AL 1 ) 1 83 I I I . A N A L Y S I S OF V A R I A N C E OF TOTAL DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N THE G A S T R O C N E M I U S MUSCLE OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR TWO WEEKS ( T R I A L 1) . 184 IV. A N A L Y S I S OF V A R I A N C E OF DNA, RNA AND P R O T E I N C O N C E N T R A T I O N IN ONE GRAM OF L I V E R OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR TWO WEEKS (TRIAL 1) . . . T85 V. A N A L Y S I S OF V A R I A N C E OF TOTAL DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N THE L I V E R S OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR TWO WEEKS ( T R I A L 1) 186 VI. A N A L Y S I S OF V A R I A N C E OF TOTAL L I V E R DNA, RNA AND P R O T E I N CONTENT PER 100G. BODY WEIGHT OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR TWO W.EEKS ( T R I A L 1) . . . 187 V I I . A N A L Y S I S OF' V A R I A N C E OF AVERAGE BODY WEIGHT G A I N , FEED I N T A K E , G A S T R O C N E M I U S MUSCLE WEIGHT, L I V E R WEIGHT AND L I V E R WEIGHT PER 100G. BODY WEIGHT OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR FOUR WEEKS ( T R I A L 1) , 188 V I I I . A N A L Y S I S OF V A R I A N C E OF DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N ONE GRAM OF G A S T R O C N E M I U S MUSCLE OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR FOUR WEEKS '('iTs.i AL 11 ) 189 IX. A N A L Y S I S OF V A R I A N C E OF TOTAL DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N THE G A S T R O C N E M I U S MUSCLE OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR FOUR WEEKS ( T R I A L 1). 190 x I TABLE PAGE X. A N A L Y S I S OF V A R I A N C E OF DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N ONE GRAM OF L I V E R OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR FOUR WEEKS ( T R I A L 1) . . • 191 XI. A N A L Y S I S OF V A R I A N C E OF TOTAL DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N THE L I V E R S OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR FOUR WEEKS (TRIAL 1) 192 X I I . A N A L Y S I S OF V A R I A N C E OF TOTAL L I V E R DNA, RNA AND P R O T E I N CONTENT PER 100G. BODY WEIGHT OF C H I C K S FED GRADED L E V E L S OF L Y S I N E FOR FOUR WEEKS (TRIAL 1) . , 1 93 X I I I . A N A L Y S I S OF V A R I A N C E OF G A S T R O C N E M I U S MUSCLE WEIGHT AND TOTAL MUSCLE DNA, RNA AND P R O T E I N CONTENT I N C H I C K S FED ADEQUATE AND E X C E S S L Y S I N E FROM ONE DAY TO 14 DAYS OF AGE ( T R I A L 2) 194 XIV. A N A L Y S I S OF V A R I A N C E OF L I V E R WEIGHT AND TOTAL L I V E R DNA, RNA AND P R O T E I N CONTENT I N C H I C K S FED ADEQUATE AND E X C E S S L Y S I N E FROM ONE DAY TO 14 DAYS OF AGE ( T R I A L 2) 1 95 XV. A N A L Y S I S OF V A R I A N C E OF G A S T R O C N E M I U S MUSCLE WEIGHT, AND TOTAL MUSCLE DNA, RNA AND P R O T E I N CONTENT I N C H I C K S FED ADEQUATE L Y S I N E AND E X C E S S L Y S I N E WITH A R G I N I N E FROM 14 TO 28 DAYS OF AGE ( T R I A L 2) 196 XVI. A N A L Y S I S OF V A R I A N C E OF L I V E R WEIGHT AND TOTAL L I V E R DNA, RNA AND P R O T E I N CONTENT I N C H I C K S FED ADEQUATE L Y S I N E AND E X C E S S L Y S I N E WITH A R G I N I N E FROM 14 TO 28 DAYS OF AGE ( T R I A L 2) 197 XVII. A N A L Y S I S OF V A R I A N C E OF AVERAGE BODY WEIGHT G A I N AND FEED I N T A K E OF C H I C K S FED D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y AND Q U A L I T Y FOR TWO AND FOUR WEEKS ( T R I A L 3) 198 X V I I I . A N A L Y S I S OF V A R I A N C E OF G A S T R O C N E M I U S MUSCLE WEIGHT AND TOTAL MUSCLE DNA, RNA AND P R O T E I N I N C H I C K S FED D I E T A R Y P R O T E I N S D I F F E R E N T I N QUANTTTY AND Q U A L I T Y FOR TWO WEEKS ( T R I A L 3) 199 XIX. A N A L Y S I S OF V A R I A N C E OF L I V E R WEIGHT AND TOTAL L I V E R DNA, RNA AND P R O T E I N I N C H I C K S FED D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y AND Q U A L I T Y FOR TWO WEEKS ( T R I A L 3) 200 x I I TABLE PAGE XX.t. A N A L Y S I S OF V A R I A N C E OF G A S T R O C N E M I U S MUSCLE WEIGHT AND TOTAL MUSCLE DNA, RNA AND P R O T E I N I N C H I C K S FED D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y AND Q U A L I T Y FOR FOUR WEEKS ( T R I A L 3) 201 XXI. A N A L Y S I S OF V A R I A N C E OF L I V E R WEIGHT AND TOTAL L I V E R DNA, RNA AND P R O T E I N I N C H I C K S FED D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y AND Q U A L I T Y FOR FOUR WEEKS ( T R I A L 3) 202 XXII. A N A L Y S I S OF V A R I A N C E OF P R O T E I N I N ONE GRAM OF MIXED T I B I A M U S C L E S , R A D I O A C T I V I T Y OF MUSCLE P R O T E I N S ( D P M / G . T I S S U E ) AND S P E C I F I C A C T I V I T Y OF MUSCLE P R O T E I N S (DPM/MG. P R O T E I N ) FROM C H I C K S FED GRADED 14 L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - C ( T R I A L 4) 203 X X I I I . A N A L Y S I S OF V A R I A N C E OF RNA I N ONE GRAM OF MIXED T I B I A M U S C L E S , R A D I O A C T I V I T Y RECOVERY ( D P M / G . P R O T E I N / P E R C E N T DOSE) AND R A D I O A C T I v I T Y / M G . RNA OF THE MUSCLES OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - ^ C ( T R I A L 4) . . 204 XXIV. A N A L Y S I S OF V A R I A N C E OF P R O T E I N I N ONE GRAM OF L I V E R , R A D I O A C T I V I T Y OF L I V E R P R O T E I N S (DPM/G. T I S S U E ) AND S P E C I F I C A C T I V I T Y OF L I V E R P R O T E I N S (DPM/MG. P R O T E I N ) FROM C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH LYS I NE-U- 1 4C ( T R I A L 4) 205 XXV. A N A L Y S I S OF V A R I A N C E OF TOTAL L I V E R P R O T E I N AND RNA FROM C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - 1 4 C ( T R I A L 4) 206 XXVI. A N A L Y S I S OF V A R I A N C E OF RNA I N ONE GRAM OF L I V E R , R A D I O A C T I V I T Y RECOVERY ( D P M / G . P R O T E I N / P E R C E N T - D O S E ) AND RAD i OACT i v i TY/MG . RNA OF THE L I V E R S OF CHI:CKS FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - 1 4 C ( T R I A L 4) 207 XXVII. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y I N MUSCLE HOMOGENATE ( D P M / G . T I S S U E ) , RNA E X T R A C T ( D P M / G . T I S S U E ) , RNA EXTRACT E X P R E S S E D AS A P E R C E N T A G E OF R A D I O A C T I V I T Y I N MUSCLE HOMOGENATE, AND S P E C I F I C A C T I V I T Y (DPM/MG. RNA) OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH URACIL-U- C ( T R I A L 5) 208 X I I I TABLE PAGE XXVIII. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y I N L I V E R HOMOGENATE (DPM/G. T I S S U E ) , RNA E X T R A C T ( D P M / G . T I S S U E ) , RNA E X T R A C T E X P R E S S E D AS A P E R C E N T A G E OF R A D I O A C T I V I T Y I N L I V E R HOMOGENATE, AND S P E C I F I C A C T I V I T Y (DPM/MG. RNA) OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH URACIL-U- C ( T R I A L 5) 209 XXIX. A N A L Y S I S OF V A R I A N C E OF AVERAGE BODY WEIGHT G A I N , FEED I N T A K E , AND R A D I O A C T I V I T Y RECOVERY I N CO2 FROM C H I C K S FED GRADED L E V E L S OF L Y S I N E AND WITH THREONINE AND A R G I N I N E ( T R I A L 6) 210 XXX. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y RECOVERY I N ACID I N S O L U B L E P R O T E I N S , ACID S O L U B L E P R O T E I N S , AND ACID I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N THE T I B I A MUSCLES OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - C ( T R I A L 7) 211 XXXI. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y RECOVERY I N A C I D I N S O L U B L E P R O T E I N S , ACID S O L U B L E P R O T E I N S , AND ACID I N S O L U B L E P R O T E I N S R E L A T I V E TO ACID S O L U B L E P R O T E I N S I N THE L I V E R S OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - ^ 4 C ( T R I A L 7) 212 XXXII. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y RECOVERY I N ACID I N S O L U B L E P R O T E I N S , ACID S O L U B L E P R O T E I N S , AND ACID I N S O L U B L E P R O T E I N S R E L A T I V E TO ACID S O L U B L E P R O T E I N S I N THE T I B I A MUSCLES OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH THREONINE-U-^ 4C ( T R I A L 7) 213 X X X I I I . A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y RECOVERY I N ACID I N S O L U B L E P R O T E I N S , ACID S O L U B L E P R O T E I N S , AND A C I D I N S O L U B L E P R O T E I N S R E L A T I V E TO ACID S O L U B L E P R O T E I N S I N THE L I V E R S OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH THREONINE-U-^ 4C ( T R I A L 7) . . . . . • 214 XXXIV. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y RECOVERY I N R E S P I R A T O R Y CO2, ACID I N S O L U B L E P R O T E I N S , ACID S O L U B L E P R O T E I N S , AND ACID I N S O L U B L E P R O T E I N S R E L A T I V E TO ACID S O L U B L E P R O T E I N S I N THE T I B I A MUSCLES OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E - U - 1 4 C ( T R I A L 8) 215 M irv, TABLE PAGE XXXV. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y RECOVERY I N ACID I N S O L U B L E P R O T E I N S , ACID S O L U B L E P R O T E I N S , AND ACID I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N THE L I V E R S OF C H I C K S . F E D GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH L Y S I N E -( T R I A L 8) •U-14C 216 XXXVI. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y RECOVERY I N R E S P I R A T O R Y COJJ, ACID I N S O L U B L E P R O T E I N S , ACID S O L U B L E P R O T E I N S , AND ACID I N S O L U B L E P R O T E I N S R E L A T I V E TO ACID S O L U B L E P R O T E I N S I N THE T I B I A MUSCLES OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH THREONINE-U- C ( T R I A L 8) 217 XXXVII. A N A L Y S I S OF V A R I A N C E OF R A D I O A C T I V I T Y RECOVERY I N A C I D I N S O L U B L E P R O T E I N S AND A C I D S O L U B L E P R O T E I N S , AND ACID I N S O L U B L E P R O T E I N S R E L A T I V E TO ACID S O L U B L E P R O T E I N S I N THE L I V E R OF C H I C K S FED GRADED L E V E L S OF L Y S I N E AND I N J E C T E D WITH THREON 1NE-U- 1 4C ( T R I A L 8) 218 XXXVIII. A N A L Y S I S OF V A R I A N C E OF AVERAGE BODY WEIGHT G A I N AND FEED INTAKE OF C H I C K S FED GRADED L E V E L S OF L Y S I N E WITH THREONINE ( T R I A L 8) 219 X^ V V LIST OF APPENDIX FIGURES F I G U R E PAGE I . CALF THYMUS DNA STANDARD CURVE 220 I | . YEAST RNA STANDARD CURVE 221 I I I . YEAST RNA STANDARD CURVE CORRECTED FOR P E P T I D E S . . . 222 IV. B O V I N E SERUM A L B U M I N STANDARD CURVE . . . . . . . . . 223 V. C^ 4 CHANNELS RATIO QUENCH C O R R E C T I O N CURVE. 224 VI. C^ 4 CHANNELS RATIO QUENCH C O R R E C T I O N CURVE 225 XV I ACKNOWLEDGEMENTS THE AUTHOR W I S H E S TO E X P R E S S H I S G R A T I T U D E TO CANADA DEPARTMENT OF A G R I C U L T U R E AND THE U N I V E R S I T Y OF B R I T I S H C O L U M B I A RESEARCH COMMITTEE FOR F I N A N C I A L S U P P O R T OF T H I S R E S E A R C H . THE A C C O M O D A T I O N P R O V I D E D BY DR. W. D. KI.TTS, CHAIRMAN OF THE DEPARTMENT OF POULTRY S C I E N C E , I S G R A T E F U L L Y ACKNOWLEDGED. S P E C I A L THANKS ARE DUE TO DR. D. B. BRAGG, A S S O C I A T E PROFESSOR OF POULTRY S C I E N C E , UNDER WHOSE S U P E R V I S I O N , I N T E R E S T AND ENCOURAGE-MENT T H I S S T U D Y WAS MADE. THE AUTHOR D E E P L Y A P P R E C I A T E S THE U S E F U L S U G G E S T I O N S , WHICH WERE V I T A L IN THE P R E P A R A T I O N OF T H I S D I S S E R T A T I O N , MADE BY THE MEMBERS OF THE GRADUATE C O M M I T T E E , P R O F E S S O R B. E. MARCH, DR. R. C. F I T Z S I M M O N S , DR. W. D. K I T T S OF THE DEPARTMENT OF POULTRY S C I E N C E AND DR. R. M. BEAMES OF THE DEPARTMENT OF ANIMAL S C I E N C E . THANKS ARE E X T E N D E D TO THE T E C H N I C A L S T A F F OF THE DEPARTMENT OF POULTRY S C I E N C E FOR T H E I R A S S I S T A N C E . THE MANAGEMENT A S S I S T A N C E G I V E N BY MR. MEL HUDSON AND THE P E R S O N N E L OF THE P O U L T R Y FARM IS ACKNOWLEDGED WITH A P P R E C I AT I ON. THE AUTHOR I S I N D E B T E D TO H I S WI F E OLUBUKOLA FOR HER E N D U R A N C E , P A T I E N C E AND U N F A I L I N G ENCOURAGEMENT D U R I N G THE I N V E S T I G A T I O N AND FOR T Y P I N G THE D R A F T OF T H I S D I S S E R T A T I O N . THE AUTHOR W I S H E S TO D E D I C A T E T H I S D I S S E R T A T I O N TO H I S DAUGHTER OLUTOKUNKO FOR HER C O O P E R A T I O N . XV I I INTRODUCTION THE P R I M A R Y F U N C T I O N OF P R O T E I N S I N THE D I E T IS TO S U P P L Y A M I N O A C I D S FOR GROWTH, R E P R O D U C T I O N AND M A I N T E N A N C E . E F F I C I E N T BODY P R O T E I N S Y N T H E S I S R E Q U I R E S THAT D I E T A R Y A M I N O A C I D S BE I N P R O P E R P R O P O R T I O N S . THE E S S E N T I A L A M I N O A C I D S , FOR E X A M P L E , MUST BE S U P P L I E D IN THE D I E T OF A M O N O G A S T R I C A N I M A L ( B I R D ) S I N C E T H E S E A M I N O A C I D S CANNOT BE S Y N T H E S I Z E D OR THE RATE OF S Y N T H E S I S I S I N S U F F I C I E N T TO MEET THE NEED OF THE A N I M A L . HOWEVER, THE N O N - E S S E N T I A L A M I N O A C I D S CAN BE S Y N T H E S I Z E D FROM A D I E T A R Y S OURCE OF O R G A N I C N I T R O G E N S U P P L I E D BY P R O T E I N . P R A C T I C A L D I E T S ARE F O R M U L A T E D TO C O N T A I N ADEQUATE P R O T E I N R E Q U I R E D BY THE M O N O G A S T R I C A N I M A L . TWO OR MORE P R O T E I N S MAY BE C O M B I N E D AND ONE OR MORE AMINO A C I D S S U P P L E M E N T E D TO THE D I E T TO O B T A I N THE ADEQUATE L E V E L . S E L E C T I O N OF P R O T E I N C O M B I N A T I O N S IS O F T E N MADE FROM A NUMBER OF NATURAL P R O T E I N S D E P E N D I N G ON Q U A L I T Y , COST AND A N I M A L P R O D U C T I V I T Y R E S U L T I N G FROM THE P R O T E I N S S E L E C T E D . MOST T E C H N I Q U E S USED I N THE E V A L U A T I O N OF P R O T E I N Q U A L I T Y ARE BASED ON GROWTH R E S P O N S E , N I T R O G E N B A L A N C E OR A M I N O A C I D C O N C E N T R A T I O N S , TWO T R A D I T I O N A L METHODS, B I O L O G I C A L V A L U E AND P R O T E I N E F F I C I E N C Y R A T I O , R E M A I N P O P U L A R . THE FORMER M E A S U R E S R E T E N T I O N OF A B S O R B E D N I T R O G E N . NET P R O T E I N U T I L I Z A T I O N , A M O D I F I C A T I O N OF B I O L O G I C A L VALUE, M E A S U R E S R E T E N T I O N OF N I T R O G E N B A S E D ON I N T A K E RATHER THAN THAT A B S O R B E D . BOTH ARE B A S E D , HOWEVER, UPON D I F F E R E N C E S IN C A R C A S S N I T R O G E N BETWEEN THE 1 T E S T GROUP AND A GROUP FED NO P R O T E I N . THE P R O T E I N E F F I C I E N C Y R A T I O ( W E I G H T G A I N PER W E I G H T OF P R O T E I N E A T E N ) I S L E S S L A B O R I O U S , S I N C E ONLY GROWTH NEED BE M E A S U R E D , BUT A L I N E A R F U N C T I O N O F T E N I S NOT O B T A I N E D . MEASUREMENTS OF N I T R O G E N B A L A N C E ARE U S E F U L BUT V A L U E S O B T A I N E D R E P R E S E N T ONLY AN A L G E B R A I C SUM OF G A I N S AND L O S S E S W I T H I N THE V A R I O U S C E L L S AND T I S S U E S . A NEW A P P R O A C H I S NOW I N D I C A T E D , BASED ON R E C E N T KNOWLEDGE OF THE B I O C H E M I C A L R E L A T I O N S H I P S OF T I S S U E D E O X Y R I B O N U C L E I C A C I D (DNA) AND R I B O N U C L E I C A C I D (RNA) C O N T E N T TO P R O T E I N B I O S Y N T H E S I S (YOUNG AND A L E X I S , 1968). THE N U T R I T I O N A L I N F L U E N C E OF T H E S E R E L A T I O N S H I P S WAS R E V E A L E D BY THE I N V E S T I G A T I O N S OF MUNRO AND C L A R K (1960) WHO FOUND THAT THE RNA C O N T E N T OF NORMAL L I V E R C E L L S OF R A T S V A R I E D WITH THE I N G E S T E D AMOUNT OF P R O T E I N AND THE A M I N O A C I D P R O F I L E OF THE P R O T E I N . S U B S E Q U E N T I N V E S T I G A T I O N S BY A L L I S O N AND CO-WORKERS (1961, 1962, 1963, 1964) I N D I C A T E D THAT DNA AND RNA C O N T E N T OF RAT L I V E R AND M U S C L E T I S S U E S I S AN I N T I M A T E F U N C T I O N NOT ONLY OF P R O T E I N R E S E R V E S I N V A R I O U S BODY T I S S U E S BUT A L S O OF THE N U T R I T I V E V A L U E OF THE D I E T A R Y P R O T E I N S . |N A D D I T I O N TO T H E S E C HANGES IN T I S S U E C O M P O S I T I O N , A M I N O A C I D D E F I C I E N C Y , E.G. T R Y P T O P H A N , MUNRO (1968); T H R E O N I N E AND I S O L E U C I N E , PRONCZUK _ET _A_L. (1970), WAS O B S E R V E D TO RETARD P R O T E I N S Y N T H E S I S ON P O L Y R I B O S O M E S C A U S I N G A D E G R A D A T I O N OF P O L Y R I B O S O M E S AND AN ACCUMU-L A T I O N OF F R E E R I B O S O M E S . LAMAR, J R . AND I MM (1971 ) A L S O DEMONSTRATED THAT E X C E S S A M I N O A C I D S S U C H AS V A L I N E , I S O L E U C I N E , H I S T I D I N E , A S P A R T I C A C I D , M E T H I O N I N E AND P H E N Y L A L A N I N E ADDED TO AN IN V I TRO A M I N O A C I D I N C O R P O R A T I N G S Y S T E M D E R I V E D FROM W E A N L I N G RAT B R A I N , I N H I B I T E D THE I N C O R P O R A T I O N OF AMIN O A C I D S INTO P R O T E I N . THE I N H I B I T I O N OCCURRED AT THE P O I N T OF A M I N O A C I D A C T I V A T I O N . THE F O R E G O I N G O B S E R V A T I O N S S U P P O R T THE S U G G E S T I O N THAT C E L L U L A R OR ORGAN CONTENT OF DNA, RNA AND P R O T E I N M I G H T S E R V E AS A C R I T E R I O N FOR E V A L U A T I N G D I E T A R Y P R O T E I N S (A L L I S ON _ET _AL . 1964). A SECOND I M P O R T A N T P O I N T E M E R G E S FROM THE F A C T THAT B I R D S E L I M I N A T E N I T R O G E N O U S COMPOUNDS V I A THE PATHWAYS OF P U R I N E M E T A B O L I S M , THE SAME PATHWAYS USED FOR THE M E T A B O L I S M OF N U C L E O T I D E S THAT MAY BE C H A N N E L E D INTO RNA S Y N T H E S I S . THE P O I N T I S THAT M E T A B O L I C PRODUCTS OF AMIN O A C I D S COULD I N F L U E N C E M E T A B O L I S M OF N U C L E I C A C I D S IN THE PATHWAYS. C O N S I D E R A T I O N OF THE P R E V I O U S R E P O R T S A L R E A D Y D E S C R I B E D AND THE F A C T THAT R E L A T I V E L Y FEW S T U D I E S ON N U C L E I C A C I D M E T A B O L I S M I N THE C H I C K E N HAVE B E E N R E P O R T E D PROMPTED THE P R E S E N T I N V E S T I G A T I O N . " T H E S T U D Y WAS D E S I G N E D TO E L U C I D A T E THE I M P A C T OF D I E T A R Y P R O T E I N AND L Y S I N E ON THE M E T A B O L I S M OF N U C L E I C A C I D S , P R O T E I N AND A M I N O A C I D S IN GROWING C H I C K S . DNA, RNA AND P R O T E I N C O N C E N T R A T I O N IN C H I C K L I V E R AND M U S C L E WAS D E T E R M I N E D UNDER D I F F E R E N T D I E T A R Y R E G I M E S . THE E F F E C T S OF D I E T A R Y L Y S I N E ON THE S Y N T H E S I S AND D E G R A D A T I O N OF T I S S U E P R O T E I N AND ON THE S Y N T H E S I S OF T I S S U E RNA WERE M E A S U R E D . THE M E T A B O L I C F A T E OF L Y S I N E WITH D I F F E R E N T I N T A K E S OF D I E T A R Y L Y S I N E WAS 14„ E L U C I D A T E D . THE O X I D A T I O N AND I N C O R P O R A T I O N OF C FROM T H R E O N I N E -14 14 U- C TO CCV, AND INTO T I S S U E P R O T E I N S R E S P E C T I V E L Y IN C H I C K S FED D I E T S C O N T A I N I N G A D E F I C I E N T , A DEQUATE AND E X C E S S L E V E L OF L Y S I N E WERE A L S O I N V E S T I G A T E D . T H R E O N I N E WAS S E L E C T E D S I N C E I T WAS NOTED TO BE THE SECOND L I M I T I N G A M I N O A C I D I N A WHEAT-BASED D I E T . LITERATURE REVIEW C E L L U L A R B A S I S FOR CHANGES I N BODY C O M P O S I T I O N D E O X Y R I B O N U C L E I C A C I D (DNA) C ONTENT OF D I P L O I D N U C L E I IN I N T E R P H A S E HAS B E E N SHOWN TO BE A C O N S T A N T Q U A N T I T Y FOR A L L THE C E L L S OF A G I V E N A N I M A L ( M L R S K Y AND R L S , 1951; THOMSON _E_T _AL. 1953 AND V E N D R E L Y , 1955). BASED ON T H I S O B S E R V A T I O N , I T HAS B E E N PROPOSED (THOMSON _ET _AL. 1953; DOUNCE _ET _AL. 1950; WATERLOW AND CO-WORKERS 1956, 1960; C A M P B E L L AND K O S T E R L I T Z , 1950) THAT THE DNA C O N C E N T R A T I O N OF A G I V E N T I S S U E BE USED AS A P O I N T OF R E F E R E N C E TO DEMONSTRATE ANY C HANGES I N THE C O N C E N T R A T I O N OF OTHER C E L L U L A R C O N S T I T U E N T S INDUCED BY THE E X P E R I M E N T A L M A N I P U L A T I O N OF THE A N I M A L . IN THE F I E L D OF N U T R I T I O N , THE C O N S T A N C Y OF DNA C O N C E N T R A T I O N IN THE L I V E R AND OTHER ORGANS HAS B E E N Q U E S T I O N E D IN THE C A S E OF RATS THAT WERE FED P R O T E I N - D E F I C I E N T D I E T S . SOME I N V E S T I G A T O R S WHO S T U D I E D THE A V E R A G E DNA C O N T E N T ( U U G / N U C L E U S ) OF THE I S O L A T E D L I V E R C E L L N U C L E U S UNDER C O N D I T I O N S OF P R O T E I N D E F I C I E N C Y , HAVE NOT B E E N A B L E TO SHOW ANY S I G N I F I C A N T I N C R E A S E I N N U C L E A R DNA C O N T E N T (THOMSON ET A L . 1953; MCINDOE AND D A V I D S O N , 1952; FUKUDA AND S I B A T A N I , 1953; C A M P B E L L AND K O S T E R L I T Z , 1947, 1948, 1952). ON THE OTHER HAND H I S T O C H E M I C A L S T U D I E S HAVE R E V E A L E D A S I G N I F I C A N T I N C R E A S E IN THE A V E R A G E DNA C O N T E N T OF THE N U C L E U S ( E L Y AND R O S E , 1951; LECOMTE AND DE SMUL, 1952). I N C O N S I S T E N T O B S E R V A T I O N S HAVE B E E N R E P O R T E D ON TOTAL L I V E R 4 HOMOG E NATE DNA. ALL I S 0 N _ET _AL . (1963) AND H A I D E R AND T A R V E R (1969) FOUND THAT THE DNA CONTENT. OF THE L I V E R WAS U N A F F E C T E D BY D I E T A R Y P R O T E I N . OTHER WORKERS (UMANA, 1965; COOPER, 1953; MUNTWYLER ET A L . 1950 AND V I L L E L A , 1952), HOWEVER, R E P O R T E D S I G N I F I C A N T I N C R E A S E S I N THE A V E R A G E DNA C O N T E N T OF THE L I V E R N U C L E U S AND I N THE DNA C O N T E N T OF THE L I V E R OF RATS FED P R O T E I N - D E F I C I E N T D I E T S . S Q U I B B (1968) A L S O O B S E R V E D THAT E X C E S S D I E T A R Y L Y S I N E C A U S E D F L U C T U A T I O N I N THE DNA C O N T E N T OF C H I C K L I V E R AND M U S C L E . GROWTH ( I N C R E A S E I N BODY W E I G H T ) OF RHODE I S L A N D RED C O C K E R E L S WAS ACCOUNTED FOR BY I N C R E A S E S IN M U S C L E WEIGHTS (MONTGOMERY ET A L . 1964). THE V I E W WAS B A S E D ON T H E I R O B S E R V A T I O N THAT M U S C L E WEIGHT: BODY-WEIGHT R A T I O OF C O C K E R E L S , F I R S T U N D E R N O U R I S H E D FOR 26 WEEKS AND THEN R E H A B I L I T A T E D , WAS THE SAME AS OF W E L L - F E D C O N T R O L S . THE O B S E R V A T I O N A L S O C O N F I R M S THE REPORT THAT S K E L E T A L M U S C L E S MAKE UP ABOUT 50$ OF TOTAL BODY W E I G H T AND THE M A J O R I T Y OF E D I B L E MEAT Y I E L D I N THE C H I C K E N ( M c N A L L Y AND S P I C K N A L L , 1959, A , B ) . P R E V I O U S R E P O R T S S T R O N G L Y S U P P O R T THE V I E W THAT M U S C L E GROWTH I N THE GROWING C H I C K DOES NOT MEAN AN I N C R E A S E I N THE NUMBER OF M U S C L E F I B R E S . S M I T H (1963), FOR E X A M P L E , NOTED THAT THE M U S C U L A R I S  S A R T O R I US A R E A AND THE M U S C U L A R I S S A R T O R I US C E L L A R I A FROM C H I C K S I N C R E A S E D AT THE SAME RATE (14 TO 15 F O L D ) FROM H A T C H I N G TO 10 WEEKS OF A G E . ON THE B A S I S OF THE O B S E R V A T I O N AND FROM H I S T O L O G I C A L S T U D I E S , SMITH (1963) CONCLUDED THAT THE I N C R E A S E I N C E L L NUMBER ( F I B R E S ) I N S K E L E T A L M U S C L E T I S S U E OF THE C H I C K OCCURRED B E F O R E H A T C H I N G AND I N C R E A S E I N M U S C L E C E L L S I Z E ( D I A M E T E R ) OCCURRED P R I M A R I L Y D U R I N G THE P O S T - H A T C H I N G P E R I O D . THIS C O N C L U S I O N C O N F I R M S P R E V I O U S R E S U L T S O B T A I N E D FROM S E V E R A L M A M M A L I A N S P E C I E S THAT D I F F E R E N T I A T I O N OF S K E L E T A L M U S C L E T I S S U E I S C O M P L E T E D P R I O R TO B I R T H I N MAMMALS AND THAT P O S T - N A T A L GROWTH OF MU S C L E MUST BE A T T R I B U T E D TO AN I N C R E A S E I N THE S I Z E OF C E L L S E X I S T I N G AT B I R T H (MACCALLUM, 1898; MCMEEKAN, 1940-41; EL I OT J_T _AL . 1943; AND MEARA, 1947). HOWEVER, THERE ARE R E P O R T S THAT THE NUMBER OF MU S C L E F I B R E S DOES I N C R E A S E A F T E R B I R T H OR H A T C H I N G . C H I A K U L A S AND PAULY (1965) O B S E R V E D THAT THE NUMBER OF F I B R E S W I T H I N A M U S C L E BUNDLE I N C R E A S E D D U R I N G E M B R Y O N I C GROWTH U N T I L T H R E E WEEKS P O S T P A R T U M . MONTGOMERY _ET _AL. (1964) A L S O R E P O R T E D THAT TOTAL NUMBER OF F I B R E S I N THE S A R T O R I U S M U S C L E S I O F THE C H I C K I N C R E A S E D BY A F A C T O R OF TWO FROM ONE DAY TO 28 WEEKS OF A G E . THE R E S U L T S THAT THERE IS AN I N C R E A S E I N THE NUMBER OF M U S C L E C E L L S D U R I N G E M B R Y O N I C GROWTH AND NOT A F T E R B I R T H OR H A T C H I N G DOES NOT I M P L Y THAT N U C L E A R D I V I S I O N DOES NOT OCCUR IN THE S K E L E T A L M U S C L E OF THE GROWING A N I M A L S I N C E S T R I A T E D M U S C L E F I B R E S ARE M U L T I - N U C L E A T E D . MUSCLE F I B R E GROWTH WAS DEMONSTRATED TOOCCUR BY I N C R E M E N T S I N THE NUMBER OF N U C L E I PER F I B R E AS WELL AS BY I N C R E M E N T S IN C Y T O P L A S M I C MASS P E R N U C L E U S ( E N E S C O AND L.EBLOND, 1962). M O S S _ E T _ A L . (1964) A L S O O B S E R V E D THAT THE NUMBER OF N U C L E I I N THE B R E A S T M U S C L E S OF THE C H I C K I N C R E A S E D 13 TO 20 FOLD AT 28 DAYS A F T E R H A T C H I N G . MOSS AND L E B L O N D (1970) HAVE A L S O I N D I C A T E D THAT THE P R O L I F E R A T I O N OF N U C L E I R E S U L T E D FROM D I V I S I O N OF S A T E L L I T E C E L L N U C L E I AND F U S I O N OF THE DAUGHTER N U C L E I INTO THE M Y O F I B R E S . N U C L E A R D I V I S I O N I N THE RAT WAS O B S E R V E D TO P R O G R E S S U N T I L 8 TO 9 WEEKS OF AGE A F T E R WHICH GROWTH I N THE RAT M U S C L E WAS BY AN I N C R E A S E I N P R O T E I N C ONTENT ONLY (GORDON _ET _AL. 1 966, AND C H E E K _ET _AL . 1 968). I N C R E A S E I N T I S S U E DNA C O N T E N T I N A GROWING A N I M A L I S A R E F L E C T I O N OF AN I N C R E A S E I N C E L L NUMBER ( H Y P E R P L A S I A ) I N THE L I V E R AND N U C L E A R D I V I S I O N I N THE M U S C L E . UNDER NORMAL C O N D I T I O N , GROWTH DOES NOT OCCUR I N THE MATURED A N I M A L AND C O N S E Q U E N T L Y A D U L T T I S S U E DNA C O N C E N T R A T I O N R E M A I N S R E L A T I V E L Y C O N S T A N T . C O N D I T I O N S S U C H AS PROLONGED S T A R V A T I O N OR P R O T E I N M A L N U T R I T I O N MAY, HOWEVER, C A U S E A D E G R A D A T I O N OF T I S S U E L A B I L E P R O T E I N R E S E R V E S (MONTGOMERY _ET _AL . 1964) TO S U P P L Y A M I N O A C I D S TO THE M E T A B O L I C POOL FOR E N E R G Y AND THE S Y N T H E S I S OF P R O T E I N S V I T A L TO L I F E . SUCH L O S S OF C Y T O P L A S M I C P R O T E I N MAY IN D E E D D E C R E A S E C E L L W E I G H T AND HENCE I N C R E A S E C E L L U L A R DNA C O N T E N T WHEN E X P R E S S E D PER U N I T W E I G H T OF T I S S U E . T H I S , OF C O U R S E , DOES NOT P R E C L U D E THE USE OF T H I S C E L L U L A R COMPONENT AS A F I X E D P O I N T OF R E F E R E N C E TO E X P R E S S THE R E S U L T S OF T I S S U E A N A L Y S I S . IMPACT OF D I E T A R Y P R O T E I N S ON T I S S U E P R O T E I N AND N U C L E I C A C I D S THE L I V E R I S R E L A T I V E L Y H I G H I N L A B I L E C Y T O P L A S M I C P R O T E I N S . MOST OF THE P R O T E I N S ARE COMPONENTS OF E N Z Y M E S Y S T E M S WHICH R I S E AND F A L L I N Q U A N T I T Y AS D I E T A R Y N I T R O G E N I N C R E A S E S OR D E C R E A S E S (WATERLOW _ET _AL. 1960). L I V E R C E L L S FROM AN IMMATURE RAT F E D ON A D I E T C O N T A I N I N G AN ADEQUATE AMOUNT OF P R O T E I N WERE O B S E R V E D TO I N C R E A S E NOT ONLY IN NUMBER BUT A L S O I N S I Z E (ENESCO AND L E B L O N D , 1962). THE P R O C E S S WAS S E V E R E L Y D E P R E S S E D BY G I V I N G A L O W - P R O T E I N D I E T , AND L I V E R C E L L S FROM T H E S E A N I M A L S HAD FAR L E S S C E L L U L A R P R O T E I N THAN THOSE FROM CONTROLS G I V E N A D I E T C O N T A I N I N G AN ADEQUATE AMOUNT OF P R O T E I N . THE D E C R E A S E D L I V E R P R O T E I N P E R C E L L I N RATS FED ON A LOW P R O T E I N D I E T WAS R E L A T E D TO A LOW RATE OF I N V I V O P R O T E I N S Y N T H E S I S (WANNEMACHER, JR. AND A L L I S O N , 1968). THE BODY AND L I V E R W E IGHTS OF RATS FED A 5$ C A S E I N D I E T FOR S E V E N DAYS AND THEN A 25$ C A S E I N D I E T FOR ONE DAY I N C R E A S E D MORE R A P I D L Y THAN THOSE OF CONTROL RATS WHICH WERE FED A 25$ C A S E I N D I E T THROUGHOUT ( H O R I E AND A S H I D A , 1971). THE I N C R E A S E I N L I V E R W E I G H T WAS A T T R I B U T E D TO AN I N C R E A S E I N T C A - I N S O L U B L E N I T R O G E N . THE RATE OF P R O T E I N S Y N T H E S I S , M E A SURED _I_N V I TRO U S I N G L I V E R S L I C E S WAS FOUND TO BE S I G N I F I C A N T L Y H I G H E R ONE DAY A F T E R THE D I E T A R Y CHANGE THAN IN CONTROL R A T S . T H I S I N C R E A S E D P R O T E I N S Y N T H E S I S WAS A L S O A C C O M P A N I E D BY ENHANCED A M I N O AC YL RNA S Y N T H E T A S E A C T I V I T Y . THE R A P I D I T Y OF THE A D J U S T M E N T OF L I V E R C O M P O S I T I O N I N D I C A T E S THAT I T MUST BE R E L A T E D I N SOME WAY TO THE I M M E D I A T E A M I N O A C I D S U P P L Y TO THE L I V E R WHICH F L U C T U A T E S W I D E L Y A F T E R E A C H MEAL B E C A U S E OF L O C A T I O N OF THAT ORGAN A S T R I D E THE P O R T A L S Y S T E M . IT I S A L S O E V I D E N T FROM P R E V I O U S R E P O R T S D E S C R I B E D ABOVE THAT ONE OF THE THREE P H A S E S OF P R O T E I N S Y N T H E S I S , A M I N O A C I D A C T I V A T I O N , I S R E S P O N S I V E TO AM I NO A C I D S U P P L Y. MUSCLE P R O T E I N L I K E THE L I V E R I S A L S O A F F E C T E D BY P R O T E I N D E F I C I E N C Y ( B ANKS _E_T _AL . 1964; HOWARTH AND B A L D WLL! N, 1 971 ; HOWARTH, 1972). D L C K E R S O N (1960) A L S O R E P O R T E D THAT TO,TAL P R O T E I N N I T R O G E N AND N O N - P R O T E I N N I T R O G E N I N C R E A S E D I N C H I C K M U S C L E WITH A G E . L I V E R P R O T E I N C O N T E N T HAS B E E N C O R R E L A T E D WITH RNA C O N T E N T BY K O S T E R L I T Z (1947) AND A L L I S O N E T _AL. (1964). IT WAS O B S E R V E D (ALL I SON _ET _AL . 1964) THAT THE RNA C O N T E N T OF THE L I V E R FROM W E A N L I N G RATS C O R R E L A T E D P O S I T I V E L Y WITH THE N U T R I T I V E V A L U E OF D I E T A R Y P R O T E I N , I N C R E A S I N G WITH A R I S E I N THE N I T R O G E N GROWTH I N D E X . TOTAL L I V E R P R O T E I N PER GRAM O F T I S S U E AND L I V E R RNA-PHOSPHOR US P E R GRAM OF T I S S U E I N C R E A S E O WITH N I T R O G E N I N T A K E B E I N G MAXIMUM I N RATS FED EGG P R O T E I N AND M I N I M U M I N THOSE FED WHEAT G L U T E N . THE P R O T E I N / RNA RA?TIO WAS C O N S T A N T , S U G G E S T I N G A C L O S E C O R R E L A T I O N BETWEEN T O T A L RNA I N THE L I V E R AND P R O T E I N B I O S Y N T H E S I S . T H I S MEANS THAT RNA, L I K E P R O T E I N C O N S T I T U E N T S OF THE L I V E R , R E S P O N D S TO A M I N O A C I D S U P P L Y . THE PROTEIN/DNA AND RNA/DNA R A T I O S I N D I C A T E THE S I Z E OF C E L L S . I N C R E A S E I N THE R A T I O S I N D I C A T E S I N C R E A S E I N THE S I Z E OF C E L L S ( H Y P E R T R O P H Y ) . ALL I SON _ET _AL . (1962) S T U D I E D THE E F F E C T S OF N I T R O G E N I N T A K E UPON T I S S U E P R O T E I N S R E L A T I V E TO DNA I N RATS FED D I E T S C O N T A I N I N G EGG, C A S E I N , C O T T O N S E E D , AND G L U T E N P R O T E I N S . T H E S E WORKERS O B S E R V E D THAT P R O T E I N / D N A AND RNA/DNA R A T I O S I N C R E A S E D I N THE L I V E R AND M U S C L E OF YOUNG RATS WITH I N C R E A S E D N I T R O G E N I N T A K E OR N U T R I T I V E V A L U E OF D I E T A R Y P R O T E I N OR BOTH. IN THE K I D N E Y , T H E S E P A R A M E T E R S I N C R E A S E D WITH N I T R O G E N I N T A K E BUT WERE U N A F F E C T E D BY THE Q U A L I T Y OF D I E T A R Y P R O T E I N . |N THE B R A I N , HOWEVER, BOTH R A T I O S WERE I N D E P E N D E N T OF T Y P E AND Q U A L I T Y OF P R O T E I N CONSUMED OVER THE E X P E R I M E N T A L P E R I O D U S E D . V ARIOUS P R O T E I N S A V A I L A B L E FOR HUMAN AND A N I M A L C O N S U M P T I O N V A R Y G R E A T L Y I N Q U A L I T Y ( E S S E N T I A L A M I N O A C I D C O M P O S I T I O N ) . EGG P R O T E I N I S H I G H I N Q U A L I T Y WHILE WHEAT G L U T E N I N N A T U R A L L Y U N B A L A N C E D , D E F I C I E N T I N L Y S I N E ( GRAU AND K A M E I , 1950). TO A V E R T THE D E L E T E R I O U S E F F E C T S OF F E E D I N G POOR Q U A L I T Y P R O T E I N , A C O M B I N A T I O N OF P R O T E I N S OR A M I N O A C I D S S U P P L E M E N T E D D I E T S ARE US E D . THE I M P A C T OF SUCH D I E T A R Y P R O T E I N S ON GROWTH AND T I S S U E S HAS A L S O B E E N S T U D I E D . BANK _ET _AL. (1964) D E T E R M I N E D THE E F F E C T OF F E E D I N G D I E T S C O N T A I N I N G WHEAT G L U T E N , WHEAT G L U T E N S U P P L E M E N T E D WITH L Y S I N E OR WHEAT G L U T E N P L U S C A S E I N TO MALE W E A N L I N G R A T S . T H E S E WORKERS O B S E R V E D THAT THE N I T R O G E N GROWTH IN D E X OF 8.6 FOR RATS FED WHEAT G L U T E N WAS IMPROVED BY E I T H E R METHOD OF S U P P L E M E N T A T I O N TO 16.7. MUSCLE PROTEIN/DNA AND RNA/DNA R A T I O S A L S O I N C R E A S E D WITH C A S E I N OR L Y S I N E S U P P L E M E N T A T I O N . L l V E R P R O T E I N / D N A AND RNA/DNA R A T I O S I N C R E A S E D O NLY WITH C A S E I N S U P P L E M E N T A T I O N . ALL I SON _ET _AL . (1964) O B S E R V E D THAT F E E D I N G A C A S E I N D I E T (15$ P R O T E I N ) OVER A P E R I O D OF 28 DAYS TO YOUNG RATS PRODUCED THE M A X I M A L I N C R E A S E I N THE L I V E R AND M U S C L E P R O T E I N / D N A R A T I O . I N C R E A S I N G D I E T A R Y P R O T E I N TO 30$ OF THE D I E T C A U S E D ONLY L I T T L E CHANGE I N LIVE.R AND M U S C L E P R O T E I N / D N A R A T I O . |N C O N T R A S T TO T H I S O B S E R V A T I O N , WEIGHT G A I N , L I V E R AND M U S C L E P R O T E I N / D N A AND RNA/DNA R A T I O S I N C R E A S E D L I N E A R L Y AS THE Q U A N T I T Y OF WHEAT G L U T E N I N THE RAT D I E T WAS I N C R E A S E D FROM 0 TO 38$. RATS G I V E N THE TWO P R O T E I N S O U R C E S AT L E V E L S TO PRODUCE THE SAME WEIGHT G A I N , 50 G., IN 28 DAYS WERE COMPARED. THOSE G I V E N C A S E I N HAD MORE L I V E R AND M U S C L E P R O T E I N AND RNA THAN RATS FED WHEAT G L U T E N . V A N D E R M E E R S - P I RE T _ET _A_L. (1966) R E P O R T E D THAT RATS D E P R I V E D OF L Y S I N E AND T H R E O N I N E BY R E P L A C I N G D I E T A R Y P R O T E I N WITH G L U T E N AND M A I Z E S T A R C H HAD LOWER L I V E R P R O T E I N (11.8$) AND RNA (71.8 UG./LOO G. L I V E R ) AS COMPARED TO 17.5$ P R O T E I N AND 81.6 UG.RNA/L00 G. L I V E R I N THE CONTROL RATS R E C E I V I N G 18$ C A S E I N . L L V E R DNA C O N C E N T R A T I O N I N L Y S I N E AND T H R E O N I N E D E P R I V E D RATS WAS S I M I L A R TO THAT I N THE CON T R O L S 11 (24.4 AND 21.9 UG./100 G. L I V E R R E S P E C T I V E L Y ) . THE SAME I M P A C T I S I N F L I C T E D ON T I S S U E S WHETHER A D I E T A R Y P R O T E I N I S FED AS I N T A C T P R O T E I N OR I N THE FORM OF A M I N O A C I D M I X T U R E , TERAOKA (1969), FOR E X A M P L E , DEMONSTRATED NO S I G N I F I C A N T D I F F E R E N C E S I N P R O T E I N AND N U C L E I C A C I D S C O NTENTS IN L I V E R , G A S T R O C N E M I U S M U S C L E , K I D N E Y , T E S T I S AND HEART FROM RATS FED A 20$ C A S E I N D I E T OR RATS FED AN A M I N O A C I D M I X T U R E S I M U L A T I N G THE P A T T E R N OF C A S E I N . D E P R E S S I O N I N FOOD I N T A K E AND GROWTH R E T A R D A T I O N O F T E N R E S U L T FROM F E E D I N G A N I M A L S _AD L I B I T U M WITH D I E T S C O M P L E T E L Y D E F I C I E N T I N A S I N G L E E S S E N T I A L A M I N O A C I D . DURAND AND P E R I O T (1969) R E P O R T E D THAT R E D U C I N G D I E T A R Y L I P I D S AND C A R B O H Y D R A T E S D E C R E A S E D E N E R G Y I N T A K E BY R A T S ; AND C A U S E D A D E C R E A S E OF TOTAL L I V E R DNA, P R O T E I N AND RNA BY 13, 12 AND 17$, R E S P E C T I V E L Y . C H E E K AND H L L L (1970), HOWEVER, O B S E R V E D NO CHANGE OR A S L I G H T I N C R E A S E I N THE M U S C L E P R O T E I N / D N A R A T I O COMPARED TO WELL FED C O N T R O L . T H E S E WORKERS S U G G E S T E D THAT C A L O R I E D E F I C I E N C Y ACTS P R I M A R I L Y ON M U S C L E DNA S Y N T H E S I S WHILE P R O T E I N D E F I C I E N C Y A C T S P R I M A R I L Y BY R E D U C I N G M U S C L E P R O T E I N S Y N T H E S I S . FOOD R E S T R I C T I O N HAS B E E N SHOWN TO HAVE A MORE S E V E R E E F F E C T ON M U S C L E DNA A C C U M U L A T I O N THAN ON P R O T E I N A C C U M U L A T I O N ( E L L I O T T AND CHEEK, 1968). HOWARTH (1972) A L S O O B S E R V E D THAT A D I E T C O N T A I N I N G 6$ P R O T E I N FED TO W E A N L I N G RATS I N H I B I T E D DNA A C C U M U L A T I O N BUT P E R M I T T E D SOME M U S C L E P R O T E I N A C C U M U L A T I O N . SOME DNA AND P R O T E I N A C C U M U L A T E D AT R A T E S L E S S THAN NORMAL WITH 12$ P R O T E I N , BUT THE D I E T I N H I B I T E D A C C U M U L A T I O N OF DNA MORE S E V E R E L Y THAN THAT OF P R O T E I N . A C C U M U L A T I O N OF P R O T E I N , RNA AND DNA I N THE M U S C L E P R O G R E S S I V E L Y D E C R E A S E D AS THE D I E T A R Y S T R E S S I N C R E A S E D FROM A M I L D P R O T E I N D E F I C I E N C Y (18$ P R O T E I N D I E T ) TO A S E V E R E D E F I C I E N C Y (6$ P R O T E I N D I E T ) . IT A P P E A R S FROM S T U D I E S BY CHEEK _ET _AL. (1970) AND HOWARTH (1972) THAT BOTH E N E R G Y ( C A L O R I E ) AND P R O T E I N D E F I C I E N C Y ACT ON M U S C L E DNA S Y N T H E S I S . THERE :|:S A L S O E V I D E N C E S U G G E S T I N G THAT BOTH T I S S U E P R O T E I N AND RNA ARE A F F E C T E D BY E N E R G Y R E S T R I C T I O N . I N C R E A S I N G E N E R G Y I N T A K E FROM 850 TO 1700 K C A L . PER SQ. M. OF BODY S U R F A C E A R E A BY A D D I N G C A R B O H Y D R A T E OR FAT TO A STANDARD D I E T ADEQUATE I N P R O T E I N , I MPROVED N I T R O G E N B A L A N C E AND L I V E R P R O T E I N C O N T E N T OF RATS (MUNRO _ET _AL. 1953). WHEN THE D I E T WAS A DEQUATE I N P R O T E I N , AN I N C R E A S E I N E N E R G Y I N T A K E OF 1000 K C A L . C A U S E D A R I S E OF 20 TO 30$ I N TOTAL RNA/LIVER. N U T R I T I O N A L R E G U L A T I O N OF T I S S U E P R O T E I N AND RNA M E T A B O L I S M IN ORDER TO C L A S S I F Y R E G U L A T O R Y R E S P O N S E S A F F E C T I N G P R O T E I N M E T A B O L I S M , I T CAN BE C O N S I D E R E D THAT F R E E A M I N O A C I D S W I T H I N THE BODY FOLLOW T H R E E T Y P E S OF M E T A B O L I C PATHWAY: (1) USED FOR P R O T E I N S Y N T H E S I S ; (2) E M P L O Y E D TO S Y N T H E S I Z E V A R I O U S COMPOUNDS OF LOW M O L E C U L A R W E I G H T SUCH AS C R E A T I N E AND N O N - E S S E N T I A L A M I N O A C I D S ; (3) UNDERGO D E G R A D A T I O N THROUGH THE PATHWAYS OF A M I N O A C I D C A T A B O L I S M . THESE C O M P E T I N G U S E S OF A M I N O A C I D S ARE I N E Q U I L I B R I U M , SO THAT AN A L T E R A T I O N I N ONE ROUTE OF D I S P O S A L I S F R E Q U E N T L Y C O M P E N S A T E D BY R E C I P R O C A L C H A N G E S I N OTHER PATHWAYS. FOR E X A M P L E , THE A D M I N I S T R A T I O N OF H Y D R A Z I N E TO RATS I M P E D E S A M I N O A C I D D E G R A D A T I O N BY I N H I B I T I N G T R A N S A M I N A T I O N ( M C C O R M I C K AND S N E L L , 1961). T H I S P R O D U C E S AN I N C R E A S E I N THE F R E E A M I N O A C I D L E V E L S I N THE T I S S U E S AND I N THE C A S E OF THE L I V E R I T HAS B E E N D E M ONSTRATED THAT P R O T E I N S Y N T H E S I S I S AUGMENTED 13 (AMENTA AND JOHNSTON, 1963). |N THE A S S E S S M E N T OF P R O T E I N N E E D S , TWO Q U E S T I O N S ARE O B V I O U S : (1) WHETHER WITH I N C R E A S I N G I N T A K E S OF P R O T E I N , T H E S E M E T A B O L I C P A T H -WAYS A D A P T L I N E A R L Y TO THE AMOUNT OF A M I N O A C I D S CONSUMED, OR WHETHER THERE IS A S UDDEN I N C R E A S E I N C A T A B O L I S M WHEN I N T A K E S OF P R O T E I N ( A M I N O A C I D S ) I N E X C E S S OF R E Q U I R E M E N T S ARE R E A C H E D ; (2) WHETHER A D A P T A T I V E R E A C T I O N S TO A M I N O A C I D S S U P P L Y OCCUR M A I N L Y D U R I N G THE P E R I O D OF A B S O R P T I O N OF THE MEAL OR D E V E L O P MORE SLOWLY IN R E L A T I O N TO A CHANGE I N G E N E R A L L E V E L OF P R O T E I N I N T A K E . |N C O N S I D E R I N G S U C H Q U E S T I O N S , IT SHOULD BE REMEMBERED THAT P R O T E I N S Y N T H E S I S OCCURS A C T I V E L Y I N MANY P A R T S OF THE BODY, WHEREAS C E R T A I N PATHWAYS I N THE C A T A B O L I S M OF E S S E N T I A L A M I N O A C I D S ARE R E S T R I C T E D TO THE L I V E R , AN E X C E P T I O N B E I N G THE BRANCHED CHA INED-AM I NO A C I D S WHICH ARE M A I N L Y DEGRADED I N THE M U S C L E ( M L L L E R , 1962). C O N S E Q U E N T L Y , V A R I A T I O N S I N A M I N O A C I D S U P P L Y ARE L I K E L Y TO EVOKE D I F F E R E N T R E S P O N S E S I N D I F F E R E N T T 1 S S U E S . THE L I V E R , FOR E X A M P L E , I S S U B J E C T E D TO AN E X T E N S I V E I N C R E A S E I N A M I N O A C I D S U P P L Y D U R I N G THE A B S O R P T I V E P E R I O D A F T E R E A C H MEAL C O N T A I N I N G P R O T E I N . THE F R E E A M I N O A C I D S I N THE P O R T A L BLOOD ROSE S E V E R A L FOLD OVER THOSE I N THE S Y S T E M I C C I RC ID IL A T I ON ( D E N T O N AND E L V E H J E M , 1954). ELWYN (1969) C A R R I E D OUT S T U D I E S TO D E T E R M I N E I F THE L I V E R P L A Y S A S I G N I F I C A N T S E L E C T I V E A C T I O N ON THE AMOUNTS OF A M I N O A C I D S THAT E N T E R THE G E N E R A L C I R C U L A T I O N . U S I N G DOGS WITH C A N N U L A S I M P L A N T E D I N THE P O R T A L V E I N , THE S P L E N I C A R T E R Y , R E P R E S E N T I N G A R T E R I A L BLOOD G O I N G INTO THE L I V E R , AND THE H E P A T I C V E I N AS I T L E A V E S THE L I V E R , HE WAS A B L E TO MONITOR C O N T I N U O U S L Y THE E X C H A N G E OF A M I N O A C I D S A C R O S S THE L I V E R D U R I N G A 24 HOUR P E R I O D . A F T E R F E E D I N G A L A R G E MEAL OF MEAT TO E X P E R I M E N T A L DOGS, I T WAS SHOWN THAT 57$ OF THE A B S O R B E D A M I N O N WAS C O N V E R T E D TO UREA AS I T P A S S E D THROUGH THE L I V E R , 6$ L E F T THE L I V E R AS P L A S M A P R O T E I N S , AND ONLY 23$ E N T E R E D THE G E N E R A L C I R C U L A T I O N AS F R E E A M I N O A C I D S . THE R E M A I N I N G 14$ WAS P R E S U M A B L Y R E T A I N E D AS HE PAT IC " P R O T E I N. A L A R G E P R O P O R T I O N OF THE I N T A K E OF B R A N C H E D - C H A I N A M I N O A C I D S P A S S E D INTO THE S Y S T E M I C C I R C U L A T I O N S I N C E T H E S E WERE DEGRADED M A I N L Y I N THE P E R I P H E R A L T I S S U E S . T H I S S U G G E S T S THAT A D A P T I V E R E A C T I O N S I N THE L I V E R P L A Y AN I M P O R T A N T ROLE I N U T I L I Z A T I O N OF D I E T A R Y P R O T E I N . F L E C K _ET _AL. (1965) E X A M I N E D THE R E S P O N S E OF P R O T E I N S Y N T H E S I S I N THE L I V E R D U R I N G THE I N F L U X OF A M I N O A C I D S . F A S T I N G RATS WERE FED BY S T O M A C H - T U B E WITH N U T R I T I O N A L L Y C O M P L E T E OR T R Y P T O P H A N D E F I C I E N T M I X T U R E S OF A M I N O A C I D S AN HOUR B E F O R E K I L L I N G . M I C R O S O M E S WERE I S O L A T E D FROM THE L I V E R S OF THE A N I M A L S AND WERE I N C U B A T E D WITH 1 4 „ C - L E U C I N E AND THE C O - F A C T O R S N E C E S S A R Y FOR C E L L - F R E E P R O T E I N S Y N T H E S I S . THE M I C R O S O M E S P R E P A R E D FROM RATS FED WITH THE A M I N O A C I D M I X T U R E L A C K I N G T R Y P T O P H A N WERE 20$ L E S S E F F I C I E N T I N I N C O R P O R A T I N G L E U C I N E INTO P E P T I D E S THAN WERE M I C R O S O M E S P R E P A R E D FROM RATS G I V E N THE N U T R I T I O N A L L Y C O M P L E T E A M I N O A C I D M I X T U R E . A N I M A L S FED THE A M I N O A C I D M I X T U R E D E F I C I E N T I N T R Y P T O P H A N AN HOUR B E F O R E K I L L I N G A L S O HAD FEWER H E A V Y P O L Y S O M E S AND MORE MONOSOMES AND D I S O M E S THAN HAD RATS G I V E N THE N U T R I T I O N A L L Y C O M P L E T E A M I N O A C I D M I X T U R E . A R E D U C T I O N I N C Y T O P L A S M I C P R O T E I N S HAS S E E N C O R R E L A T E D WITH A D E C R E A S E I N RNA/DNA R A T I O . |T I S A L S O KNOWN THAT THE RNA/DNA R A T I O I S I N F L U E N C E D BY THE P A T T E R N OF A M I N O A C I D S ; THE MORE D E F I C I E N T THE P A T T E R N THE L E S S RNA I N THE C E L L S (MUNRO AND C L A R K , 1960 AND A L L I S O N E_T _A_L . 1962). HOWARTH (1971 ) A L S O R E P O R T E D THAT BOTH THE AMOUNT AND THE A C T I V I T Y OF RNA D E T E R M I N E S THE RATE OF M U S C L E P R O T E I N S Y N T H E S I S . THIS C O R R E L A T I O N I N THE C HANGES I N T I S S U E P R O T E I N AND RNA E M P H A S I Z E S THAT THERE MUST BE SOME FORM OF I N T E G R A T I O N I N THE M E T A B O L I C R E S P O N S E OF C E L L U L A R COMPONENTS TO A M I N O A C I D S U P P L Y . E X P E R I M E N T A L F I N D I N G S OF S T U D I E S D E S I G N E D TO E L U C I D A T E THE N U T R I T I O N A L R E G U L A T I O N OF C E L L U L A R P R O T E I N AND RNA M E T A B O L I S M WERE S U M M A R I Z E D BY MUNRO (1968). THE O V E R A L L P I C T U R E S U G G E S T S THAT THE I N F L U X OF A M I N O A C I D S INTO THE L I V E R A F T E R A MEAL S T I M U L A T E S P R O T E I N S Y N T H E S I S ; R E D U C E S P R O T E I N BREAKDOWN AND E N T R A I N S MORE R I B O S O M E S INTO P O L Y S O M E S , SO THAT FEWER BECOME D E G R A D E D . WHEN A B S O R P T I O N OF A M I N O A C I D C E A S E S , THE P R O C E S S E S ARE R E V E R S E D , R E S U L T I N G I N A C C E L E R A T E D BREAKDOWN BOTH OF P R O T E I N AND OF RNA I N THE C E L L . T H I S RNA - P R O T E I N CONTROL M E C H A N I S M I S C O M P A T I B L E WITH THE P A R A L L E L C H A N G E S I N C O N C E N T R A T I O N OF T H E S E C E L L U L A R COMPONENTS AT V A R I O U S L E V E L S OF P R O T E I N I N T A K E AND S U P P O R T E D BY THE F I N D I N G S OF WANNEMACHER ET A L . (1968) THAT H E P A T I C P R O T E I N S Y N T H E S I S I S R E G U L A T E D BY D I E T A R Y A M I N O A C I D S BY A F F E C T I N G THE A B I L I T Y OF P O L Y R I B O S O M E S TO S Y N T H E S I Z E P R O T E I N AND BY A F F E C T I N G THE C O N C E N T R A T I O N OF C Y T O P L A S M I C R I B O S O M E S . WUNNER _ET _AL. (1 966) O B S E R V E D THAT D E F I C I E N C Y OF T R Y P T O P H A N I N THE RAT D I E T C A U S E D D I S A G G R E G A T I O N OF C E L L U L A R P O L Y S O M E S . UN THE B A S I S OF T H I S E F F E C T ON L I V E R P O L Y S O M E S I T WAS S U G G E S T E D THAT P R O T E I N S Y N T H E S I S ON THE P O L Y S O M E S WAS R E T A R D E D BY T R Y P T O P H A N D E F I C I E N C Y C A U S I N G AN A C C U M U L A T I O N OF F R E E R I B O S O M E S A W A I T I N G A T T A C H M E N T TO M E S S E N G E R RNA. RECENT S T U D I E S ON THE I M P A C T OF T H R E O N I N E AND I S O L E U C I N E D E F I C I E N C Y ON RAT L I V E R P O L Y S O M E S WERE R E P O R T E D BY P R O N C Z U K ET A L . (1970). DURING THE I N V E S T I G A T I O N , RATS WERE FED AD L I B I T U M LOW L E V E L S OF C R Y S T A L L I N E A M I N O A C I D S L I M I T I N G IN I S O L E U C I N E OR T H R E O N I N E OR THE SAME A M I N O A C I D M I X T U R E WITH T H R E O N I N E OR I S O L E U C I N E ADDED TO I T . L L V E R P O L Y S O M E P R O F I L E S WERE E X A M I N E D A F T E R THE D I E T S WERE FED TO RATS FOR ONE OR FOUR HOURS AND FROM 10 HOURS TO 14 DAYS I N THE F I R S T AND SECOND T R I A L R E S P E C T I V E L Y . BOTH S T U D I E S R E V E A L E D THAT L I V E R S FROM RATS FED T H R E O N I N E OR I S O L E U C I N E S U P P L E M E N T E D D I E T SHOWED MORE P O L Y S O M E AND FEWER MONOSOMES AND D I S O M E S THAN L I V E R S FROM RATS R E C E I V I N G D I E T S L I M I T I N G I N E I T H E R OF THE TWO A M I N O A C I D S . T H E S E F I N D I N G S I N D I C A T E THAT THE P O L Y S O M E P R O F I L E S I N THE L I V E R OF THE RAT CAN BE I N F L U E N C E D BY D I E T A R Y A M I N O A C I D S OTHER THAN T R Y P T O P H A N , P R O V I D E D THAT A S U F F I C I E N T L Y S E V E R E A M I N O A C I D D E F I C I E N C Y CAN BE C R E A T E D . ANOTHER I M P O R T A N T P O I N T E MERGES FROM T H E S E E A R L I E R S T U D I E S (WUNNER _ET _AL. 1 966 AND PRONCZ UK _ET _AL . 1970). THAT I S , THE R A P I D RATE AT WHICH A M I N O A C I D D E F I C I E N C Y A F F E C T S L I V E R P O L Y S O M E P R O F I L E S I M P L I E S THAT A V A I L A B I L I T Y OF RI BOS OMAL RNA I S A MAJOR F A C T O R I N L I V E R P R O T E I N B I O S Y N T H E S I S BOTH I N THE SHORT AND LONG TERM R E G U L A T I O N , HOWARTH (1972), HOWEVER, S U G G E S T E D THAT THE A V A I L A B I L I T Y OF R I B O S O M A L RNA WAS A MAJOR F A C T O R ONLY IN THE LONG TERM R E G U L A T I O N OF M U S C L E P R O T E I N B I O S Y N T H E S I S . SHORT TERM CONTROL OF L I V E R P R O T E I N B I O S Y N T H E S I S MAY R E F E R TO THE S Y N T H E S I S OF L I V E R E N Z Y M E S . FOR E X A M P L E , S C H I M K E (1968) R E P O R T E D THAT MANY E N Z Y M E S ARE S T A B I L I Z E D BY T H E I R S U B S T R A T E S SO THAT THE E N Z Y M E P R O T E I N I S C A T A B O L I ZED AT A SLOWER R A T E . C O N S E Q U E N T L Y , A M I N O A C I D S A B S O R B E D A F T E R A P R O T E I N MEAL W I L L REDUCE THE BREAKDOWN OF THE NUMEROUS E N Z Y M E S I N V O L V E D I N A M I N O A C I D D E G R A D A T I O N AND A L S O S T I M U L A T E L I V E R P R O T E I N S Y N T H E S I S . BOTH OF T H E S E F A C T O R S WOULD C O N T R I B U T E TO THE R E S P O N S E OF F E E D I N G D I E T A R Y P R O T E I N C A U S I N G A R A P I D I N C R E A S E I N L I V E R P R O T E I N C O N T E N T . T H I S P R O C E S S I S MUCH MORE I N T E N S E I N THE L I V E R FLIHAN I N THE M U S C L E , B E C A U S E OF I T S D I R E C T A C C E S S TO THE H I G H C O N C E N T R A T I O N S OF A B S O R B E D A M I N O A C I D S . C O N S E Q U E N T L Y , THE L I V E R SHOWS A C O N S I D E R A B L Y H I G H E R C O N T E N T OF L A B I L E P R O T E I N S THAN DO OTHER T I S S U E S (MUNRO, 1964). THE M E C H A N I S M OF P R O T E I N TURNOVER WITH D I E T WAS I L L U S T R A T E D BY THE S T U D I E S OF GAN AND J E F F A Y (1967) I N WHICH THE Y MEASUREO THE C O N T R I B U T I O N TO THE I N T R A C E L L U L A R POOL OF A M I N O A C I D S R E L E A S E D THROUGH L I V E R P R O T E I N BREAKDOWN. A N I M A L S C O N S U M I N G FOOD SHOWED A 50$ C O N T R I B U T I O N TO THE POOL OF L Y S I N E FROM R E C Y C L I N G OF P R O T E I N S W I T H I N THE C E L L , WHEREAS A SHORT P E R I O D OF F A S T I N G R A I S E D T H I S TO 90$ OF THE A M I N O A C I D S U P P L Y . T I S S U E P R O T E I N S Y N T H E S I S AND D E G R A D A T I O N I N R E L A T I O N TO D I E T A R Y P R O T E I N D I E T A R Y P R O T E I N A F F E C T S SOME T I S S U E S MORE R A P I D L Y AND E X T E N S I V E L Y THAN OTHERS (WATERLOW _ET AJ_. 1960). THE B R A I N I S MOST R E S I S T A N T TO D E P L E T I O N IN P R O T E I N S , FOLLOWED BY THE K I D N E Y , THE M U S C L E AND F I N A L L Y THE L I V E R (W A I N I 0 _ET _AL . 1 959J A L L I S O N AND F l T Z P A T R I C K , 1960). S C H I M K E (1964), U S I N G T R A C E R T E C H N I Q U E S , SHOWED THAT 50$ OF THE L I V E R P R O T E I N WAS R E P L A C E D FROM THE D I E T A R Y SOURCE W I T H I N 4 TO 5 DAYS AND 76$ I N 20 D A Y S . I T WAS A L S O R E P O R T E D THAT T I S S U E P R O T E I N S F A L L I NTO TWO C A T E G O R I E S , N A M E L Y , S T A B L E AND L A B I L E P R O T E I N S ( A D D I S ET AL . 1936, K O S T E R L I T Z AND C A M P B E L L , 1945 AND K O S T E R L I T Z , 1947). THE IM P A C T OF D I E T A R Y P R O T E I N I S P A R T I C U L A R L Y MARKED IN THE L I V E R B E C A U S E THE ORGAN HAS H I G H L A B I L E C Y T O P L A S M I C P R O T E I N S . MOST OF THE CYTO-P L A S M I C P R O T E I N S ARE COMPONENTS O F ENZYME S Y S T E M S WHICH R I S E AND F A L L I N Q U A N T I T Y AS D I E T A R Y N I T R O G E N I N C R E A S E S OR D E C R E A S E S . I F T H I S L A B I L E P R O T E I N I S B E I N G C O N T I N U A L L Y B U I L T UP AND BROK E N DOWN AT A G R E A T E R RATE THAN THE TURNOVER RATE OF THE P R O T E I N I N A N Y G I V E N T I S S U E , I T W I L L DEPEND TO A L A R G E E X T E N T ON THE D I E T A R Y S T A T E OF THE A N I M A L . I N A D D I T I O N , THE RATE OF P R O T E I N S Y N T H E S I S W I L L A L S O BE E X P E C T E D TO VA R Y UNDER D I F F E R E N T D I E T A R Y P R O T E I N R E G I M E . I S O T O P I C A L L Y L A B E L E D A M I N O A C I D S HAVE B E E N A D M I N I S T E R E D TO E X P E R I M E N T A L A N I M A L S I N ORDER TO D E T E R M I N E THE TURNOVER R A T E S OF BODY P R O T E I N S . TWO H Y P O T H E S E S HAVE B E E N P R O P O S E D BY T A R V E R (1954) AND S T E I NBERG _ET _AL . (1956) TO J U S T I F Y THE R A D I O I S O T O P E S T U D I E S . THE F I R S T H Y P O T H E S I S I S THAT AN A M I N O A C I D I S R E L E A S E D OR E X C H A N G E D FROM W I T H I N A P R O T E I N M O L E C U L E BY C O M P L E T E P R O T E I N D E G R A D A T I O N . THE P R O T E I N I S R E S Y N T H E S I ZED FROM A M I N O A C I D S BASED ON THE P O S T U L A T E D " T E M P L A T E M E C H A N I S M " OF P R O T E I N S Y N T H E S I S . SUCH A M E C H A N I S M HAS G E N E R A L L Y B E E N ASSUMED I N THE I N T E R P R E T A T I O N OF TURNOVER S T U D I E S OF S I N G L Y L A B E L E D P R O T E I N S AND MOST M U L T I P L E L A B E L E D P R O T E I N S . THE I M P L I C A T I O N I S THAT A S I N G L E I S O T O P I C A L L Y L A B E L E D A M I N O A C I D S P E C I E S W I T H I N A P R O T E I N HAS A H A L F - L I F E EQUAL TO THAT OF A L L THE OTHER A M I N O A C I D S P E C I E S W I T H I N THE P R O T E I N AND HENCE OF THE P R O T E I N I T S E L F . THE SECOND H Y P O T H E S I S IS THAT OF P A R T I A L D E G R A D A T I O N , W H E R E I N THE P R O T E I N I S H Y D R O L Y Z E D INTO AND R E S Y N T H E S I Z E D FROM P E P T I D E S . T H I S I S B A S E D ON THE P O S T U L A T E D " P E P T I D E M E C H A N I S M " OF P R O T E I N S Y N T H E S I S . BOTH H Y P O T H E S E S HAVE R E C E I V E D TREMENDOUS S U P P O R T THROUGHOUT THE L I T E R A T U R E . THE F I R S T H Y P O T H E S I S I S S U P P O R T E D BY THE F O L L O W I N G 1 4 S T U D I E S : THE WORK OF A S K O N A S _EJ_ _AL. (1955) SHOWED THAT V A L I N E - 4 - C 1 4 AND L Y S I N E - 1 - C HAD THE SAME R E S P E C T I V E S P E C I F I C A C T I V I T I E S E I T H E R I N C A S E I N AND B-LAC T O G L 0 B U L I N OR I N E A C H OF 27 P E P T I D E S I S O L A T E D FROM P A R T I A L HYDROLYSA'/TES OF T H E S E P R O T E I N S . S l M P S O N AND V E L I C K (1954) AND HEIMBERG AND V E L I C K ( (1 954) O B S E R V E D THAT THE R A T I O S OF 14 14 THE S P E C I F I C A C T I V I T I E S OF G L Y C I N E - 1 - C, A L A N I N E - 1 - C, P H E N Y L -14, ' „ 14„ A L A N I N E - 3 - C AND L Y S I N E - 2 - C R E M A I N E D C O N S T A N T AND WERE THE SAME IN R A B B I T M U S C L E A L D O L A S E AND P H O S P H 0 R Y L A S E . M c F A R L A N E (1956) ALSO 14, 131, SHOWED THE TURNOVER OF C AND | I N DOUBLY L A B E L E D P L A S M A P R O T E I N S WAS E Q U A L . THE SECOND H Y P O T H E S I S I S S U P P O R T E D BY THE F O L L O W I N G O B S E R V A T I O N S : A N F I N S E N AND CO-WORKERS (1951, 1952, 1954) FOUND UNEQUAL L A B E L I N G OF THE A M I N O A C I D R E S I D U E S I N O V A L B U M I N , I N S U L I N AND R I B O N U C L E A S E . M A R G E N AND T A R V E R (1957) O B S E R V E D THAT THE H A L F - L I F E OF AND ^ | |N 14 DOUBLY L A B E L E D P L A S M A P R O T E I N S WERE UN E Q U A L . V A L I N E - 1 - C AND 35 13 14 M E T H I O N I N E - S OR V A L I N E - 1 - C AND L E U C I N E - 1 - C, R E S P E C T I V E L Y , I N D O U B L Y - L A B E L E D RAT A L B U M I N A L S O HAD D I F F E R E N T TURNOVER R A T E S (WLGGANS _ET _AL . 1 956 AND R U M S F E L D _E_T _AL . 1957). THE D I S T R I B U T I O N P A T T E R N OF R A D I O A C T I V E L Y L A B E L E D A M I N O A C I D S INTO THE P R O T E I N AND F R E E A M I N O A C I D F R A C T I O N OF T I S S U E S I S WELL E S T A B L I S H E D . T A R V E R AND MORSE (1948) DEMONSTRATED THAT WHEN M E T H I O N I N E L A B E L E D WITH R A D I O A C T I V E S U L F U R WAS FED TO R A T S , THE C O N C E N T R A T I O N OF R A D I O A C T I V E S U L F U R A F T E R 24 HOURS WAS G R E A T E S T I N THE P R O T E I N S OF THE I N T E S T I N A L MUCOSA AND D E C R E A S E D I N THE ORDER OF K I D N E Y , P L A S M A , L I V E R , S P L E E N AND B R A I N . THE SKLIN, H A I R AND M U S C L E R E S I D U E HAD A V E R Y LOW C O N C E N T R A T I O N OF R A D I O A C T I V E S U L F U R . I N T R A V E N O U S A D M I N I S T R A T I O N , HOWEVER, GAVE THE F O L L O W I N G P A T T E R N OF 35e I N C O R P O R A T I O N OF S I N T H E I R ORDER OF D E C R E A S I N G A C T I V I T Y : I N T E S T I N A L MUCOSA, P A N C R E A S , S P L E E N , K I D N E Y , P L A S M A , L I V E R , T E S T I S , H E A R T , B R A I N AND M U S C L E . THE TURNOVER RATE OF T I S S U E P R O T E I N I S I N F L U E N C E D BY D I E T A R Y P R O T E I N . SOLOMON AND TARVER (1952) O B S E R V E D THAT THE RATE OF L O S S OF L A B E L FROM THE P R O T E I N I N P L A S M A , L I V E R AND K I D N E Y WAS H I G H ON A 45 OR 50$ P R O T E I N D I E T AND MUCH L E S S I N A D I E T C O N T A I N I N G 30$ OR L E S S P R O T E I N . S T E I N B O C K AND T A R V E R (1954) A L S O FOUND THAT WITH A P R O T E I N - F R E E D I E T , THE R E P L A C E M E N T RATE OF P L A S M A P R O T E I N S WAS S M A L L WHEREAS ON A H I G H P R O T E I N (25 OR 65$) D I E T I T WAS TWO OR MORE T I M E S H I G H E R . J E F F A Y AND W L N Z L E R (1958) R E P O R T E D S I M I L A R O B S E R V A T I O N S : SERUM A L B U M I N OF RATS M A I N T A I N E D ON LOW (0 OR 5$) P R O T E I N D I E T S SHOWED LONGER H A L F - L I F E AND SLOWER R E P L A C E M E N T R A T E S THAN THOSE I N RATS M A I N T A I N E D ON ADEQUATE P R O T E I N (12$). S E R U M A L B U M I N OF RATS M A I N T A I N E D ON H I G H (30 TO 40$) P R O T E I N D I E T S TURNED OVER MORE R A P I D L Y THAN A L B U M I N OF RATS M A I N T A I N E D ON ADEQUATE P R O T E I N . T H I S F A S T E R TURNOVER WAS R E F L E C T E D I N BOTH A D E C R E A S E OF A L B U M I N H A L F - L I F E AND AN I N C R E A S E R E P L A C E M E N T R A T E . HOWEVER, THE TURNOVER RATE OF THE S L A B E L L E D S E R U M G L O B U L I N WAS NOT D E P E N D E N T ON D I E T A R Y P R O T E I N . THE TURNOVER RATE OF L I V E R , TOTAL P L A S M A AND S E R U M A L B U M I N P R O T E I N I N 21 R A T S WAS A L S O O B S E R V E D TO I N C R E A S E WITH I N C R E A S I N G THE L E V E L S O F D I E T A R Y P R O T E I N BY MURAMATSU AND A S H I D A (1962). 1 5 S H E M I N AND R l T T E N B E R G (1944) SHOWED BY F E E D I N G G L Y C I N E - N THAT THE H A L F - L I F E OF L I V E R N I T R O G E N WAS ABOUT S E V E N D A Y S . p R I E D B E R G _ET _A_L . (1948) O B S E R V E D THE H A L F - L I F E IN L I V E R P R O T E I N S TO BE 3.7 DAYS 35„ BY I N J E C T I N G M E T H I O N I N E - S. HOWEVER, L I V E R P R O T E I N S HAD A H A L F - L I F E 35 OF 2.8 DAYS A F T E R F E E D I N G M E T H I O N I N E - S ( T A R V E R AND MORSE, 1948). THE A V E R A G E H A L F - L I F E OF P R O T E I N S I N SOME T I S S U E S OF RATS FED 25$ C A S E I N D I E T WERE A L S O R E P O R T E D BY MURAMATSU _ET _AL. (1963). I N T E S T I N A L MUCOSA HAD A H A L F - L I F E OF 2 TO 2.9 D A Y S , P L A S M A 3.3 TO 4.4 D A Y S , S P L E E N 4.1 D A Y S , K I D N E Y 4.1 TO 4.9 DAYS AND L I V E R 5.0 D A Y S . THE S P E C I F I C A C T I V I T Y OF L I V E R AND SE R U M P R O T E I N S FOR RATS FED A T H R E O N I N E OR T R Y P T O P H A N D E F I C I E N T D I E T R E S P E C T I V E L Y WAS FOUND TO BE H I G H E R THAN THAT FOR THE S U P P L E M E N T E D GROUP BY S I D R A N S K Y AND FARBER (1958), NIMNI AND BAVETTA (1961). THE S P E C I F I C A C T I V I T Y OF M E T H I O N I N E (SOLOMON AND T A R V E R , 1952, MURAMATSU _E_T _AL . 1963) AND L Y S I N E (HAIDER AND TARVER, 1969) WAS R E S P E C T I V E L Y FOUND TO VAR Y I N V E R S E L Y WITH THE P R O T E I N C O N T E N T OF THE D I E T , THE H I G H E S T S P E C I F I C A C T I V I T Y B E I N G FOR L I V E R P R O T E I N S FROM RATS FED A P R O T E I N - F R E E D I E T AND LOWEST FOR THOSE FED H I G H P R O T E I N D I E T . H A I D E R AND T A R V E R (1969) C A L C U L A T E D R A T E S OF L I V E R P R O T E I N S Y N T H E S I S TO BE 17.1, 23.2, 15.6 AND 26.2 MG./HOUR/200G. BODY WEIGHT; AND OF P L A S M A A L B U M I N AS 7.52, 9.55, 6.79 AND 3.73 MG./HOUR/200G. BODY WEIGHT FOR NORMAL, LOW P R O T E I N , H I G H P R O T E I N AND P R O T E I N - F R E E D I E T S , R E S P E C T I V E L Y . THE P R O T E I N D E G R A D A T I O N R A T E S I N S K E L E T A L M U S C L E HAS B E E N MEASURED ONLY I N R E C E N T Y E A R S . FR I TZ _ET _AL . (1969) F O L L O W I N G 22 1 4„ C O N T I N U O U S D I E T A R Y A D M I N I S T R A T I O N OF A C- L A B E L E D Y E A S T P R O T E I N H Y D R O L Y S A T E TO ADULT RATS O B T A I N E D A H A L F - L I F E OF 31 AND 22 DAYS FOR L A C T A T E DEHYDROGENASE - 5 AND TOTAL S O L U B L E P R O T E I N , R E S P E C T I V E L Y , I N S K E L E T A L M U S C L E . THE E F F E C T OF S T A R V A T I O N AND A P R O T E I N - F R E E D I E T ON THE S Y N T H E S I S AND C A T A B O L I S M OF S K E L E T A L M U S C L E P R O T E I N S WAS M E A S U R E D BY MLLLWARD 1 4 (1970). RATS W E I G H I N G 90 TO 100G. WERE I N J E C T E D WITH C- L A B E L E D C A R B O N A T E AND THE F R A C T I O N A L RATE OF C A T A B O L I S M WAS E S T I M A T E D FOR THE S A R C O P L A S M I C AND M Y O F I B R I L L A R F R A C T I O N OF S K E L E T A L M U S C L E BY THE DE C A Y RATE OF A S P A R T A T E AND G L U T A M A T E . THE H A L F - L I F E FOR P R O T E I N D E G R A D A T I O N WAS 3.6 AND 15.6 DAYS FOR THE S A R C O P L A S M I C AND M Y O F I B R I L L A R P R O T E I N S , R E S P E C T I V E L Y , I N YOUNG R A T S . THE R A T E S OF S Y N T H E S I S AND D E G R A D A T I O N OF M I X E D P R O T E I N S WERE A L S O M E ASURED R E C E N T L Y BY YOUNG _ET _AL. (1971) I N THE S K E L E T A L M U S C L E OF RATS A F T E R AD L I B I T U M F E E D I N G OF A LOW P R O T E I N D I E T . T H E S E WORKERS FOUND THAT THE H A L F - L I F E FOR D E G R A D A T I O N OF M I X E D M U S C L E P R O T E I N S WAS 13.1 DAYS FOR WELL N O U R I S H E D MALE RATS W E I G H I N G 75 TO 106G. AS COMPARED WITH A V A L U E OF 10.2 DAYS FOR D E P L E T E D R A T S . T I S S U E RNA S Y N T H E S I S AND D E G R A D A T I O N I N R E L A T I O N TO D E I T A R Y P R O T E I N THE I M P A C T OF D I E T A R Y P R O T E I N ON THE AMOUNT OF RNA I N THE L I V E R HAS A L R E A D Y B E E N D E S C R I B E D AS S U B J E C T TO N U T R I T I O N A L C O N T R O L . WHEN A RAT I S S T A R V E D ( D A V I D S O N , 1947 AND THOMSON _ET _AL. 1953) OR I S FED ON A P R O T E I N D E F I C I E N T D I E T ( K O S T E R L I T Z , 1947 AND M UNRO _ET _AL . 1953),. THERE I S AN I M M E D I A T E AND E X T E N S I V E L O S S OF RNA FROM THE L I V E R FOR ABOUT 2 D A Y S , A F T E R WHICH A NEW AND LOWER P L A T E A U I S A T T A I N E D . I N C O R P O R A T I O N OF C - G L Y C I N E INTO THE P U R I N E B A S E S AND C-OROTIC A C I D INTO THE P Y R I M I D I N E B A S E S OF L I V E R RNA WAS FOUND BY C L A R K _ET A L . (1957) AND MUNRO AND C L A R K (1959) TO BE C O N S I D E R A B L Y G R E A T E R WHEN A C O M P L E T E A M I N O A C I D M I X T U R E WAS G I V E N THAN WHEN A M I X T U R E L A C K I N G T R Y P T O P H A N WAS F E D . T H E S E R E S E A R C H E R S A L S O CONCLUDED THAT THE R A P I D R E D U C T I O N I N L I V E R RNA C O N T E N T D U R I N G THE F I R S T 24-28 HOUR ON A P R O T E I N - D E F I C I E N T D I E T WAS DUE TO A C C E L E R A T E D BREAKDOWN OF L I V E R RNA RATHER THAN TO R E D U C T I O N I N THE RATE OF RNA S Y N T H E S I S . THE P R E V I O U S S T U D I E S AND THE O B S E R V A T I O N OF WUNNER ET A L . (1966) THAT F E E D I N G A M I N O A C I D M I X T U R E S L I M I T I N G I N T R Y P T O P H A N TO RATS BY STOMACH TUBE C A U S E D A R E D U C T I O N I N H E A V Y A G G R E G A T E S AND AN I N C R E A S E I N D I S O M E S AND OTHER S M A L L E R P A R T I C L E S AND A D E C R E A S E I N THE S P E C I F I C 30 1 9 A C T I V I T Y OF THE S AND S RNA F R A C T I O N S LED MUNRO (1968) TO CO N C L U D E THAT THE RNA CO N T E N T OF THE L I V E R I S D E T E R M I N E D BY CH A N G E S I N RATE OF RNA BREAKDOWN C A U S E D BY V A R I A T I O N I N A M I N O A C I D S U P P L Y . THE RATE OF D E G R A D A T I O N OF RNA I N THE L I V E R S OF RATS HAS B E E N MEASURED BY S E V E R A L AUTHOR S . G E R B E R _ET _AL . (1960) E X A M I N E D THE RATE OF L O S S OF L A B E L FROM BOTH THE R I B O S E AND THE B A S E S OF L I V E R RNA FROM RATS R E C E I V I N G A NORMAL D I E T AND CONCLUDED THAT FLIHE MEAN TURNOVER T I M E OF RNA IN RAT L I V E R WAS 7.2 D A Y S ; T H I S I S E Q U I V A L E N T TO A H A L F - L I F E OF ABOUT 5 D A Y S . T H I S F I G U R E FOR TOTAL L I V E R RNA A G R E E S WITH L A T E R E S T I M A T E S OF THE H A L F - L I F E OF L I V E R R I B O S O M A L RNA (LOEB _E_T _AL . 1965, H L R S C H AND H I A T T , 1966, W L L S O N AND HOAGLAND, 1967, BLOBEL AND POTTER, 1 968 AND ENWONWU _ET _AL. 1971), WITH THE E X C E P T I O N OF ONE LOW F I G U R E OF 40 HOURS ( H A D J I O L O V , 1966). THE E F F E C T OF TO T A L S T A R V A T I O N ON THE TURNOVER I N RAT L I V E R RNA WAS E X A M I N E D BY H l R S C H AND HI ATT (1966) AND IT WAS CONCLUDED THAT THE L O S S OF RNA THAT OCCURS D U R I N G S T A R V A T I O N WAS DUE TO AN I N C R E A S E I N R I B O S O M A L RNA D E G R A D A T I O N , AS WELL AS TO R E D U C T I O N I N THE RATE OF RNA S N Y T H E S I S . ENWONWU AND MUNRO (1970) S T U D I E D I N T A K E OF P R O T E I N ON RNA TURNOVER. THE TOTAL AMOUNT OF RNA C O N T A I N E D I N THE L I V E R S OF RATS FED A F I X E D AMOUNT (15G.) OF 18$ C A S E I N D I E T ROSE BY 1$ PER DAY BETWEEN THE 6 AND 13TH DAY OF F E E D I N G WHEREAS THE A N I M A L S ON A S I M I L A R I N T A K E OF P R O T E I N - F R E E D I E T SHOWED A D A I L Y F R A C T I O N A L RNA L O S S OF -3$ P E R DAY, THE H A L F - L I F E OF R I B O S O M A L RNA WAS C A L C U L A T E D TO BE 65 HOURS ON THE 18$ C A S E I N D I E T , E Q U I V A L E N T TO A TURNOVER T I M E OF 94 HOURS, A F R A C T I O N A L D E G R A D A T I O N RATE OF 26$ OF L I V E R RNA PER DAY AND A F R A C T I O N A L RATE OF S Y N T H E S I S OF 27$ PER DAY. |N THE C A S E OF THE A N I M A L S ON THE P R O T E I N - F R E E D I E T , THE H A L F - L I F E OF L I V E R R I B O S O M A L RNA WAS 96 HOURS, E Q U I V A L E N T TO A TURNOVER T I M E OF 139 HOURS, A F R A C T I O N A L D E G R A D A T I O N RATE OF 17$ AND A F R A C T I O N A L S Y N T H E T I C RATE OF 14$ PER DAY. RNA TURNOVER D U R I N G THE T R A N S I T I O N P E R I O D FROM AN AD L I B I T U M ADEQUATE P R O T E I N I N T A K E TO A P R O T E I N - F R E E D I E T WAS E X A M I N E D BY ENWONWU AND MUNRO (1970). THE TOTAL RNA C O N T E N T OF THE L I V E R S OF RATS R E C E I V I N G 18$ P R O T E I N D I E T ROSE BY +1$ PER DAY D U R I N G THE S T U D Y . WITHDRAWAL OF P R O T E I N R E S U L T E D I N A L O S S OF L I V E R RNA, O C C U R I N G I N TWO P H A S E S . DURING THE F I R S T TWO D A Y S , THERE WAS A S H A R P R E D U C T I O N AT A F R A C T I O N A L RATE OF 10$ PER DAY. T H E R E A F T E R , THE RATE OF L O S S SLOWED TO':5$ PER DAY. RATS ON THE 18$ C A S E I N D I E T THROUGHOUT THE S T U D Y SHOWED A P R O G R E S S I V E D E C R E A S E IN TOTAL R A D I O A C T I V I T Y I N THE L I V E R RNA WITH A H A L F - L I F E OF 81 HOURS, A F R A C T I O N A L D E G R A D A T I O N RATE OF 21$ AND A S Y N T H E T I C RATE OF 22$. THE RATS THAT R E C E I V E D THE P R O T E I N - F R E E D I E T SHOWED C O N S I D E R A B L Y A C C E L E R A T E D D E G R A D A T I O N D U R I N G THE F I R S T 2 DAYS A F T E R D E L E T I O N OF P R O T E I N FROM THE D I E T WITH A H A L F -L I F E OF 38 HOURS, A F R A C T I O N A L RATE OF RNA BREAKDOWN OF 43$ ( T W I C E AS F A S T AS THE RATE OF D E G R A D A T I O N ON THE 18$ C A S E I N D I E T ) AND A S Y N T H E T I C RATE OF 33$ PER DAY. PROLONGED S T A R V A T I O N ON THE RATE OF RAT L I V E R RNA S Y N T H E S I S AND D E G R A D A T I O N (E NWONWU _E_T _AL . 1971) GAVE R E S U L T S S I M I L A R TO THOSE D E S C R I B E D A B O V E . THE F R A C T I O N A L C A T A B O L I C AND S Y N T H E T I C R A T E S WERE 14 AND 16$ PER DAY R E S P E C T I V E L Y I N FED B I R D S . D E G R A D A T I O N OF RNA I N S T A R V A T I O N WAS AT F I R S T A C C E L E R A T E D TO ABOUT 50$ PER DAY D U R I N G WHICH T I M E S Y N T H E T I C RATE WAS A L S O I N C R E A S E D , BUT A F T E R 48 HOURS OF F A S T I N G THE F R A C T I O N A L RATE OF D E G R A D A T I O N F E L L TO 10$ PER DAY. T H I S DROP I N D E G R A D A T I O N WAS A C C O M P A N I E D BY A G R E A T E R F A L L I N S Y N T H E S I S SO THAT F A S T E D A N I M A L S SHOWED A SLOW BUT C O N T I N U O U S L O S S OF L I V E R RNA. CONCOMITANT WITH THE P H A S E OF E X T E N S I V E L O S S OF RNA I N F A S T E D RATS WAS THE C O M P L E T E D I S A G G R E G A T I O N OF L I V E R P O L Y S O M E S AND A MARKED I N C R E A S E I N M E M B R A N E - F R E E R I B O S O M E S . ON THE B A S I S OF T H E S E S T U D I E S , E NWONWU _ET _AL. (1971) CONCLUDED THAT: ( A ) S Y N T H E S I S OF L I V E R RNA I S NOT D E P R E S S E D BY F A S T I N G U N T I L S I G N I F I C A N T L O S S E S OF RNA AND P O L Y S O M E S HAVE O C C U R E D j ( B ) THE E F F E C T OF F A S T I N G ON DEGRADAT I V E RATE V A R I E S WITH THE D U R A T I O N OF S T A R V A T I O N AND (c) R I B O S O M A L RNA TURNOVER I N RAT L I V E R I S I N PART R E G U L A T E D THROUGH CHANGES I N THE P O P U L A T I O N OF M E M B R A N E - F R E E R I B O S O M E S AND S U B U N I T S , THE ABUNDANCE OF WHICH I S A F F E C T E D BY FOOD D E P R I V A T I O N . RODRIGUEZ (1969) DEMONSTRATED THAT P U R I F I E D PH 5 E N Z Y M E S AND R I B O N U C L E O P R O T E I N (RNP) P A R T I C L E S FROM RATS FED WHEAT G L U T E N SHOWED A H I G H E R RATE OF L A B E L L I N G WHICH WAS I N D E P E N D E N T OF E N Z Y M E CONCEN-T R A T I O N THAN THOSE FROM C A S E I N - F E D A N I M A L S . T H E S E R E S E A R C H E R S A L S O FOUND THAT ON I N C U B A T I N G THE A C T I V A T I N G AND T R A N S F E R E N Z Y M E S E L U T E D FROM DEAE-CELLULOSE COLUMNS, P H E N O L - E X T R A C T E D S O L U B L E RNA (SRNA) AND 14 RNP P A R T I C L E S WITH L E U C I N E - C, S P E C I F I C A C T I V I T Y OF THE S Y S T E M FROM G L U T E N - F E D A N I M A L S WAS 42$ G R E A T E R . L A B E L L E D PHE N Y L - A L A N I NE , L Y S I N E AND A L A N I N E A L S O GAVE S I M I L A R I N C R E M E N T S . S T A R V A T I O N AND P R O T E I N S I N D I E T S A P P E A R TO BE TWO F A C T O R S SHOWN TO A F F E C T RNA S Y N T H E S I S AND D E G R A D A T I O N . THE I N F O R M A T I O N A V A I L A B L E C O N C E R N I N G THE I N F L U E N C E OF D I E T A R Y P R O T E I N ON RNA S Y N T H E S I S CAME M A I N L Y FROM RAT S T U D I E S . R E C E N T L Y , A SHORT S T U D Y OF THE E F F E C T OF AMI N O A C I D S ON L I V E R RNA S Y N T H E S I S I N THE C H I C K WAS R E P O R T E D ( H E B E R T _ET _AL. 1 972). TWO-WEEK-OLD C H I C K S WERE A D M I N I S T E R E D O R A L L Y WITH M I X T U R E S OF V A R I O U S K I N D S AND AMOUNTS OF AM I N O A C I D S . THE E F F E C T OF T H E S E A M I N O A C I D S ON RNA S Y N T H E S I S WAS MEASURED BY THE I N C O R P O R A T I O N 3 OF T R I T I U M FROM U R I D I N E - 5 - H INTO C H I C K L I V E R RNA FOR TWO HOURS A F T E R D O S I N G . HIGH L E V E L S OF A M I N O A C I D S (300 MG.) A D M I N I S T E R E D TO C H I C K S R E S U L T E D I N A D E C R E A S E I N RNA S Y N T H E S I S . OF THE P R E C U R S O R S OF THE P U R I N E R I N G T E S T E D ( F O L I C A C I D , G L Y C I N E , G L U T A M I N E AND A S P A R T I C AC I D ) ONLY A S P A R T I C A C I D WAS FOUND TO A F F E C T RNA S Y N T H E S I S . |T I N C R E A S E D THE RATE OF RNA S Y N T H E S I S . E F F E C T OF V A R I O U S A M I N O A C I D I M B A L A N C E S ON THE I N C O R P O R A T I O N OF 14 G L Y C I N E - 1 - C INTO RNA AND P R O T E I N WAS A L S O D E T E R M I N E D BY H E B E R T E T A L . (1972). IT WAS O B S E R V E D THAT I M B A L A N C E S OF (1) E S S E N T I A L A M I N O A C I D S P L U S 50 MG. OF G L Y C I N E , (2) E S S E N T I A L A M I N O A C I D S M I N U S T R Y P T O P H A N AND (3) E S S E N T I A L A M I N O A C I D S M I N U S M E T H I O N I N E R E S U L T E D I N S I G N I F I C A N T L Y 14 LOWER I N C O R P O R A T I O N OF G L Y C I N E - 1 - C INTO RNA THAN THE I N C O R P O R A T I O N O B T A I N E D WITH A C O M P L E T E B A L A N C E OF A M I N O A C I D S . OF A L L THE A M I N O A C I D I M B A L A N C E S T E S T E D , A D E F I C I E N C Y OF T R Y P T O P H A N HAD THE G R E A T E S T E F F E C T . M E T A B O L I S M OF S P E C I F I C AMINO A C I D S AS I N F L U E N C E D BY D I E T A R Y AMINO  A C I D CONTENT A L A R G E P R O P O R T I O N OF A D I E T A R Y A M I N O A C I D P R E S E N T I N S U B O P T I M A L AMOUNTS I S USED FOR P R O T E I N S Y N T H E S I S . AS THE D I E T A R Y S U P P L Y E X C E E D S THE A N I M A L NEEDS FOR P R O T E I N S Y N T H E S I S , I N C R E A S E D USE OF THE C A R B O N S K E L E T O N FOR A L T E R N A T I V E P R O C E S S E S SUCH AS L I P O G E N E S I S , GLUCONEO-G E N E S I S , E X C R E T I O N I N THE U R I N E OR I N C R E A S E D O X I D A T I O N TO C A R B O N D I O X I D E MAY BE E X P E C T E D . BROOKES _E_T _AL . (1972) D E V E L O P E D AND E V A L U A T E D AN O X I D A T I O N T E C H N I Q U E FOR E S T I M A T I N G THE L Y S I N E R E Q U I R E M E N T S OF THE RAT. RATS FED GRADED L E V E L S OF L Y S I N E WERE I N J E C T E D WITH RAD I O A C T I V E L Y L A B E L E D L Y S I N E V I A HEART P U N C T U R E . THE P R O D U C T I O N OF 1 4 COg WAS USED AS AN I N D E X OF THE AMOUNT OF L Y S I N E O X I D I Z E D . R E S U L T S O B T A I N E D I N D I C A T E D THAT THE O X I D A T I O N OF L Y S I N E D I D NOT I N C R E A S E M A R K E D L Y U N T I L THE D I E T A R Y L Y S I N E I N T A K E WAS I N C R E A S E D ABOVE THAT L E V E L AT WHICH A V E R A G E D A I L Y G A I N AND G A I N / F E E D WERE MAXIMUM. THE O X I D A T I O N OF A SECOND A M I N O A C I D , M E T H I O N I N E , BY RATS FED GRADED L E V E L S OF L Y S I N E WAS SHOWN TO BE I N D E P E N D E N T OF L Y S I N E I N T A K E (BROOKES _ET A_L. 1972). Y A M A S H I T A AND SAKURAI (1 966) ON THE OTHER HAND, O B S E R V E D THAT RATS D E P R I V E D OF L Y S I N E E X C R E T E D ALMOST A L L THE T H R E O N I N E FROM THE D I E T I N U R I N E , THE AMOUNT OF T H R E O N I N E E X C R E T E D D E P E N D I N G ON 28 L Y S I N E C O N T E N T I N THE D I E T . E X C R E T I O N OF T H R E O N I N E WAS P R O B A B L Y A M E C H A N I S M TO R I D THE BODY OF THE A M I N O A C I D S I N C E I T WAS FOUND TO A C C U M U L A T E IN THE P L A S M A AND WAS NOT C H A N N E L L E D INTO P R O T E I N WITH L Y S I N E OR A R G I N I N E D E F I C I E N C Y ( Z I M M E R M A N AND S C O T T , 1965). THE M E T A B O L I C E F F E C T OF I M B A L A N C E D A M I N O A C I D S WAS S T U D I E D BY Y O S H I D A _ET _AL_. (1966), HARTMAN AND K I NG (1967). T H E S E WORKERS DEMONSTRATED THAT I N C O R P O R A T I O N OF I S O T O P I C A L L Y L A B E L E D L I M I T I N G A M I N O A C I D S INTO T I S S U E P R O T E I N OF RATS WAS S T I M U L A T E D BY T H R E O N I N E OR H I S T I D I N E I M B A L A N C E D (LOW P R O T E I N ) D I E T . O X I D A T I O N OF THE L I M I T I N G A M I N O A C I D BY RATS FED THE A M I N O A C I D I M B A L A N C E D D I E T WAS A L S O AT A S I M I L A R RATE AS I N THE CONTROL RATS R E C E I V I N G WELL B A L A N C E D D I E T . THE M E T A B O L I C BREAKDOWN OF THE L I M I T I N G A M I N O A C I D S IN RATS FED A M I N O AC I D - I M B A L A N C E D D I E T S WERE O B S E R V E D TO D E C R E A S E ( W L L S O N ET AL . 1962, AND YOSH I DA _ET _AL . 1966). KEAN (1966) A L S O O B S E R V E D AN E N H A N C E M E N T OF A C T I V I T Y OF THE M E T H I O N I N E A C T I V A T I N G E N Z Y M E IN RATS FED A M E T H I ON I N E - D E F I C I E N T D I E T . RATS F O R C E - F E D A D I E T D E V O I D OF T H R E O N I N E , TO WHICH V A R I O U S R A D I O A C T I V E A M I N O A C I D S E.G. L E U C I N E , I S O L E U C I N E , AND V A L I N E WERE I N J E C T E D SHOWED A D E C R E A S E D RATE OF P R O T E I N S Y N T H E S I S I N THE S K E L E T A L M U S C L E . THE R A T E , HOWEVER, A P P E A R E D TO I N C R E A S E I N THE L I V E R ( S l D R A N S K Y AND CO-WORKERS, 1958, 1964, 1965, 1967). Y A M A S H I T A AND A S H I D A (1969) I N A S I M I L A R S T U D Y , O B S E R V E D THAT CHANGES I N THE RATE OF P R O T E I N S Y N T H E S I S DUE TO F E E D I N G OF T H R E 0 N I N E - D E F I C I E N T D I E T S AD L I B I T U M E X H I B I T E D A S I M I L A R P A T T E R N I N BOTH L I V E R AND M U S C L E , ALTHOUGH THE E F F E C T WAS MORE MARKED I N THE M U S C L E . I T WAS A L S O DEMONSTRATED THAT THE I N C O R P O R A T I O N OF L E U C I N E - C INTO P R O T E I N 29 D E C R E A S E D I N BOTH L I V E R AND M U S C L E OF RATS FED T H R E 0 N I N E - D E F I C I E N T 14, D I E T S WHEREAS I N C O R P O R A T I O N OF T H R E O N I N E - C I N C R E A S E D BY F E E D I N G THE SAME D I E T S . A G R E A T E R R E L A T I V E S P E C I F I C A C T I V I T Y OF P R O T E I N WAS O B T A I N E D FROM RATS FED THRE 0 N 1 N E - D E F I C I E N T D I E T S THAN FROM THE GROUP FED THE B A S A L D I E T . THE H I G H E R R E L A T I V E S P E C I F I C A C T I V I T Y OF P R O T E I N WAS ACCOUNTED FOR BY A G R E A T E R TURNOVER AND RE U T I L I Z A TI ON RATE OF THE T H R E O N I N E M O L E C U L E . 1 4 THE M E T A B O L I C T R A N S F O R M A T I O N S OF L Y S I N E - U - C AND T H R E O N I N E - U -14, C I N RATS C O N S U M I N G L Y S I N E - AND T H R E O N I N E - D E F I C I E N T D I E T S R E S P E C T I V E L Y WERE MEASURED BY Y A M A S H I T A AND A S H I D A (1969). THE P E R C E N T A G E R E C O V E R Y 14 14 OF C FROM L Y S I N E - U - C I N R E S P I R A T O R Y CC^, BLOOD SUGAR, L I V E R G L Y C O G E N , L I V E R L I P I D S AND C A R C A S S L I P I D S I N RATS FED L YS I N E - F R E E 14„ D I E T D E C R E A S E D C O N S I D E R A B L Y , WHEREAS THAT OF T H R E O N I N E - U - C I N T H E S E F R A C T I O N S OF RATS FED THE T H R E 0 N I N E - F R E E GROUP WAS S I M I L A R TO C O N T R O L S . 14 P E R C E N T A G E R E C O V E R Y OF C I N THE P R O T E I N F R A C T I O N R E L A T I V E TO THAT I N THE R E S P I R A T O R Y C0^ WAS A L S O H I G H E R I N THE L Y S I N E - F R E E D I E T - F E D GROUPS THAN I N THE T H R E 0 N I N E - F R E E D I E T G R O U P S . I T WAS S U G G E S T E D ON THE B A S I S OF T H E S E R E S U L T S THAT SOME HOMTJS TAT I C M E C H A N I S M O P E R A T E S I N A N I M A L S FED L Y S I N E - F R E E D I E T TO M A I N T A I N THE L E V E L OF L Y S I N E I N THE BODY COMPARED WITH THAT OF T H R E O N I N E I N THE A N I M A L BODY ON THE T H R E O N I N E - F R E E D I E T . E F F E C T OF E X C E S S I V E L E V E L S OF L Y S I N E AND T H R E O N I N E ON THE M E T A B O L I S M OF T H E S E A M I N O A C I D S IN THE RAT WAS A L S O I N V E S T I G A T E D BY Y A M A S H I T A AND A S H I D A (1971). FREE T H R E O N I N E C O N C E N T R A T I O N I N P L A S M A , L I V E R AND M U S C L E OF RATS WERE O B S E R V E D TO I N C R E A S E M A R K E D L Y DUE TO THE F E E D I N G OF E X C E S S T H R E O N I N E WHEREAS T I S S U E L Y S I N E C O N T E N T R E M A I N E D 30 R E L A T I V E L Y C O N S T A N T WHEN THE E X C E S S L Y S I N E D I E T WAS F E D . THE C A T A B O L I C 1 4 BREAKDOWN OF T H R E O N I N E - U — C TO R E S P I R A T O R Y C0^ I N RATS FED THE E X C E S S 14. T H R E O N I N E D I E T D I D NOT I N C R E A S E AS MUCH AS THAT OF L Y S I N E - U - C I N RATS FED THE E X C E S S L Y S I N E D I E T . THE E X C E S S I V E F E E D I N G OF EACH R E S P E C T I V E A M I N O A C I D R E S U L T E D I N A LOWERED R A D I O A C T I V I T Y R E C O V E R Y I N THE P R O T E I N F R A C T I O N I N C O N T R A S T TO THE E L E V A T E D R E C O V E R Y I N THE A C I D - S O L U B L E F R A C T I O N , THE E F F E C T OF THE E X C E S S T H R E O N I N E D I E T B E I N G MORE MARKED THAN THAT OF THE E X C E S S L Y S I N E D I E T . R E S U L T S OF THE R A D I O A C T I V I T Y M E A S U R E M E N T OF THE M E T A B O L I C T R A N S F O R M A T I O N OF I N T R A -14- 14-PER I T O N E A L L Y A D M I N I S T E R E D L Y S I N E - U - C AND T H R E O N I N E - U - C TO BLOOD SUGAR, L I V E R G L Y C O G E N AND' L I P I D S AND C A R C A S S L I P I D S SHOWED THAT THE 14„ C O N V E R S I O N OF T H R E O N I N E - U - C /TO C A R B O H Y D R A T E G R E A T L Y I N C R E A S E D I N RATS FED THE E X C E S S T H R E O N I N E D I E T . THE R E S U L T S A L S O S U P P O R T THE V I E W C O N C E R N I N G A H O M E O S T A T I C M E C H A N I S M O P E R A T I V E I N RATS WHICH M A I N T A I N S A C O N S T A N T BODY L E V E L OF L Y S I N E AND THE A B S E N C E OF SUCH A M E C H A N I S M TO M A I N T A I N T H R E O N I N E L E V E L S WHICH F L U C T U A T E D E P E N D I N G UPON THE F E E D I N G C O N D I T I O N S . CONTROL MECHANISM OF FEED I N T A K E D E P R E S S I O N I N FOOD I N T A K E AND C O N S E Q U E N T L Y GROWTH R E T A R D A T I O N OF GROWING A N I M A L S FED AD L I B I T U M WITH AN I M B A L A N C E D D I E T OR D I E T S D E F I C I E N T I N OR C O N T A I N I N G E X C E S S OF ONE E S S E N T I A L A M I N O A C I D HAVE B E E N SHOWN BY S A U B E R L I C H E_T AJL . (1955), H A R P E R _ET AJL . (1955), S A N A H U J A _ET _AL. (1962), LE UNG _ET _AL . 1964, ANDERSON _E_T _AL. 1969, MARCH AND WALKER (1970) AND BRAGG _ET _AL. (1971). GROWING RATS WOULD EAT A P R O T E I N - F R E E OR A B A L A N C E D P R O T E I N D I E T R A T H E R THAN AN I M B A L A N C E D D I E T WHEN O F F E R E D A C H O I C E BETWEEN THEM ( S A N A H U J A AND HARPER, 1963 AND LEUNG _ET _AL. 1968). MANY S T U D I E S HAVE B E E N C A R R I E D OUT TO E L U C I D A T E THE C A U S E OF THE D E P R E S S I O N I N FOOD I N T A K E A L R E A D Y D E S C R I B E D . S A L M O N (1958) S U G G E S T E D THAT A M I N O A C I D I M B A L A N C E REDUCED THE E F F I C I E N C Y OF U T I L I Z A -T I O N OF THE L I M I T I N G A M I N O A C I D AND N I T R O G E N . KU NT A _ET _A_L . (1958), HOWEVER, I N D I C A T E D THAT M E A S U R E M E N T S OF N I T R O G E N B A L A N C E COULD G I V E NO I N F O R M A T I O N ABOUT THE B A S I S FOR A D V E R S E E F F E C T S OF C O N S U M I N G AN 1MBALANCED D I E T . T H I S S U G G E S T I O N WAS BA S E D ON T H E I R O B S E R V A T I O N THAT A M I N O A C I D I M B A L A N C E D E P R E S S E D N I T R O G E N R E T E N T I O N NO MORE THAN D I D E Q U I V A L E N T R E S T R I C T I O N OF FOOD I N T A K E AND THE D E P R E S S I O N OF FOOD I N T A K E OCCURRED R A P I D L Y A F T E R I N G E S T I O N OF THE I M B A L A N C E D D I E T . P A L A T A B I L I T Y WAS RULED OUT AS THE C A U S E OF D E P R E S S I O N I N FOOD I N T A K E B E C A U S E RATS I N G E S T E D A B A L A N C E D D I E T AT THE SAME RATE AS AN I M B A L A N C E D D I E T B E F O R E A D E P R E S S I O N I N FOOD I N T A K E OF THE GROUP G I V E N THE I M B A L A N C E D D I E T COULD BE D E T E C T E D ( H A R P E R AND KUNTA, 1959). KUNTA AND HARPER (1961) MEASURED STOMACH E M P T I N G P E R I O D AND FOUND I T NOT TO BE D E L A Y E D BY A M I N O A C I D I M B A L A N C E . I M P A R E D P R O T E I N D I G E S T I O N WAS A L S O R E P O R T E D U N L I K E L Y TO C A U S E D E P R E S S I O N OF F E E D C O N S U M P T I O N BY HENDERSON _ET _AL. (1953). E V I D E N C E WAS A L S O P R E S E N T E D BY YOSHIDA ET A L . (1966), PENG AND HARPER (1969) THAT A M I N O A C I D S ADDED TO C R E A T E AN I M B A L A N C E D D I E T D|D NOT I N T E R F E R E WITH THE A B S O R P T I O N OF THE L I M I T I N G AM I NO AC 1D. SALMON (1954, 1958) P O S T U L A T E D THAT THE A D D I T I O N OF AN I N C O M P L E T E M I X T U R E OF A M I N O A C I D S TO A LOW P R O T E I N D I E T M I G H T I N C R E A S E THE C A T A B O L I S M OF A L L AMINO A C I D S AND HENCE R E S U L T I N I N C R E A S E D D E S T R U C T I O N OF THE MOST L I M I T I N G A M I N O A C I D I N THE D I E T . T H I S H Y P O T H E S I S R E C E I V E D L I T T L E S U P P O R T FROM THE M E T A B O L I S M OF I S O T O P I C A L L Y L A B E L L E D T R Y P T O P H A N I N RATS ( F L O R E NT I NO _ET _AL . 1962) AND S T U D I E S OF BLOOD UREA (KUNTA AND HARPER, 1961). IT WAS O B S E R V E D , HOWEVER, BY S A U B E R L I C H AND SALMON (1955) THAT U R I N A R Y E X C R E T I O N OF A M I N O A C I D S BY RATS G I V E N AN I M B A L A N C E D D I E T WAS E I T H E R NOT I N C R E A S E D OR ENHANCED V E R Y S L I G H T L Y . MOREOVER, W I L S O N _ET _AL . (1 962) FOUND NO I N D I C A T I O N OF I N C R E A S E D C A T A B O L I S M OF T R Y P T O P H A N , THE L I M I T I N G A M I N O A C I D I N THE D I E T U S E D . YOSH I DA _ET _A_L . (1966) FOLLOWED THE M E T A B O L I S M OF THE 14, L I M I T I N G A M I N O A C I D , T R A C E R Q U A N T I T I E S OF T H R E O N I N E - U - C OR J 4 , H I S T I D I N E - U - C I N RATS FED T H R E O N I N E OR H I S T I D I N E I M B A L A N C E D D I E T 1 4 LOW I N P R O T E I N . T H E Y O B S E R V E D THE SAME AMOUNT C FROM T H R E O N I N E I N CARBO N D I O X I D E , U R I N E AND F E C E S FROM A N I M A L S FED I M B A L A N C E D OR CONTROL D I E T S I N D I C A T I N G THAT THE C A T A B O L I S M OF THE L I M I T I N G A M I N O A C I D FROM F E E D I N G AN I M B A L A N C E D D I E T I S NOT E N H A N C E D . ON THE B A S I S OF T H E S E O B S E R V A T I O N S THE I N C R E A S E D A M I N O A C I D E X C R E T I O N AND C A T A B O L I S M C O N C E P T CANNOT BE USED TO E X P L A I N THE D E P R E S S I O N I N FOOD I N T A K E . PLASMA A M I N O A C I D C O N C E N T R A T I O N I S I M P L I C A T E D FOR SOME OF THE D E P R E S S I O N I N FOOD I N T A K E O C C U R I N G AS A R E S U L T OF I N G E S T I O N OF AN A M I N O A C I D I M B A L A N C E D D I E T . S A N A H U J A AND H A R P E R (1963) O B S E R V E D THAT 1 F E E D I N G TO RATS A H I S T I D I N E I M B A L A N C E D D I E T C A U S E D A D E P R E S S I O N I N FOOD I N T A K E P R E C E D E D BY A MARKED F A L L I N P L A S M A H I S T I D I N E C O N C E N T R A -T I O N AND A R I S E I N THE C O N C E N T R A T I O N OF OTHER E S S E N T I A L A M I N O A C I D S E S P E C I A L L Y T H R E O N I N E . |T I S NOT C L E A R HOW THE P L A S M A A M I N O A C I D P A T T E R N R E S U L T S I N A S I G N A L TO REDUCE F E E D I N T A K E . THERE I S A S P E C U L A T I O N , HOWEVER, THAT 33 P L A S M A A M I N O A C I D P A T T E R N MAY BE A R E F L E C T I O N OF THE CHANGE THAT OCCURS AT THE S I T E OF P R O T E I N S Y N T H E S I S ( I N THE M U S C L E OR OTHER ORGAN) AND THAT A S I G N A L I S T R A N S M I T T E D BY SOME NERVOUS OR HORMONAL MECHAN I SM. B E N E V E N G A _ET _AL. (1968) I N D I C A T E D THAT P R O T E I N S Y N T H E S I S I N THE L I V E R WAS S T I M U L A T E D W I T H I N A FEW HOURS A F T E R I N G E S T I O N OF A D I E T I M B A L A N C E D WITH R E S P E C T TO H I S T I D I N E . I S O T O P I C S T U D I E S A L S O R E V E A L E D THAT AS MUCH OR MORE OF A L I M I T I N G A M I N O A C I D ( T H R E O N I N E ) WAS IN C O R P O R A T E D INTO C A R C A S S AND T I S S U E P R O T E I N S OF RATS FED T H R E O N I N E I M B A L A N C E D D I E T COMPARED WITH CONTROLS FED THE B A L A N C E D D I E T ( YOSH I DA _ET _AL . 1966). THE ENHANCED I N C O R P O R A T I O N OF THE L I M I T I N G A M I N O A C I D INTO T I S S U E P R O T E I N S WOULD BE E X P E C T E D TO REDUCE THE AMOUNT C I R C U L A T I N G I N BODY F L U I D S AND COULD ACCOUNT FOR LOW C O N C E N T R A T I O N O B S E R V E D I N BLOOD. HARPER AND ROGERS (1965) D E V E L O P E D THE F O L L O W I N G H Y P O T H E S I S C O N C E R N I N G THE S E Q U E N C E OF E V E N T S L E A D I N G TO THE A L T E R E D P L A S M A A M I N O A C I D P A T T E R N OF RAT I N G E S T I N G AN I M B A L A N C E D D I E T . D I E T A R Y P R O T E I N I S D I G E S T E D N O R M A L L Y AND THE A M I N O A C I D S FROM THE MEAL ARE E F F I C I E N T L Y A B S O R B E D . T H I S R E S U L T S I N S U R P L U S OF A L L BUT ONE OF THE E S S E N T I A L A M I N O A C I D S I N THE POR T A L BLOOD F L O W I N G TO THE L I V E R . THE S U R P L U S OF A M I N O A C I D S S T I M U L A T E S S Y N T H E S I S OR S U P P R E S S E S BREAKDOWN OF P R O T E I N SO THAT MORE OF THE L I M I T I N G A M I N O A C I D I S R E T A I N E D I N THE L I V E R I N THE I M B A L A N C E D THAN I N THE CO N T R O L . M U S C L E P R O T E I N C O N T I N U E S TO BE S Y N T H E S I Z E D AT A NORMAL R A T E , SO THE E V E N T U A L R E S U L T I S THAT THE F R E E A M I N O A C I D P A T T E R N S OF BOTH P L A S M A AND M U S C L E BECOME S E V E R E L Y I M B A L A N C E D , R E S E M B L I N G THOSE PRODUCED BY I N G E S T I O N OF A S E V E R E L Y 34 D E F I C I E N T D I E T . SOME OF T H E S E CHANGES ARE MONITORED BY AN A P P E T I T E -R E G U L A T I N G C E N T R E AND FOOD I N T A K E I S D E P R E S S E D . AS A C O N S E Q U E N C E OF LOWER F E E D I N T A K E , THE S U P P L Y OF E N E R G Y AND A M I N O A C I D S I S REDUCED AND GROWTH I S R E T A R D E D . A S E R I E S OF S T U D Y CONDUCTED BY S L N D R A S K Y AND CO-WORKERS (1958, 1960, 1961, 1962, 1964) SHOWED THAT YOUNG RATS F O R C E F E D P U R I F I E D D I E T S D E V O I D OF S I N G L E E S S E N T I A L A M I N O A C I D S , D E V E L O P E D P A T H O L O G I C A L C H A N G E S . THE A N I M A L S WERE P R O B A B L Y LOADED WITH A M I N O A C I D S WHICH COULD NOT BE C H A N N E L L E D INTO P R O T E I N S Y N T H E S I S OR DEGRADED F A S T ENOUGH, AND HENCE THE A D V E R S E E F F E C T S . D E P R E S S I O N OF THE L I M I T I N G A M I N O A C I D IN P L A S M A AND F E E D I N T A K E R E S U L T I N G FROM F E E D I N G AN I M B A L A N C E D D I E T COULD BE A P R O T E C T I V E D E V I C E ON THE PART OF THE AN I M A L . THERE ARE I N D I C A T I O N S THAT C O N S U M P T I O N OF AN I M B A L A N C E D D I E T I S S T I M U L A T E D BY I N S U L I N I N J E C T I O N ( K U N T A AND H A R P E R , 1962), C O R T I S O L I N J E C T I O N ( L E U N G _ET _AL . 1964), L - T H Y R O X I N E I N J E C T I O N ( B E A T O N , 1 967), BY A D J U S T M E N T OF P R O T E I N : C A L O R I E R A T I O O F : T H E D I E T ( F L S H E R AND S H A P I R O , 1961), BY E X P O S U R E OF THE A N I M A L S TO A COLD E N V I R O N M E N T (KLA I N _ET _AL . 1 962, HARPER AND ROGERS, 1966) AND BY FORCE F E E D I N G (LEUNG _ E T _ A L . 1964). ALL OF T H E S E T E C H N I Q U E S R E S U L T E D IN IMPROVED GROWTH E Q U I V A L E N T TO THAT OF CONTROL A N I M A L S . I T I S U N D E R S T A N D A B L E THAT AN A N I M A L E A T S MORE OF A D I E T I N A COLD THAN A WARM E N V I R O N M E N T . THE A N I M A L E A T S MORE TO MEET I T S E N E R G Y NEEDS FOR "WARMING U P " . BUT WHY SUCH A N I M A L E A T S MORE OF AN I M B A L A N C E D THAN A B A L A N C E D D I E T I N A COLD E N V I R O N M E N T ( S A N A H U J A AND H A R P E R , 1962) I S UNKNOWN. A R I S E I N BODY T E M P E R A T U R E WAS O B S E R V E D I N C H I C K S FED A L YS I N E - D E F I C I E N T D I E T BY P AS T R 0 _ET _AL . (1969A). A D E C L I N E I N T H Y R O I D A C T I V I T Y I N R E S P O N S E TO I N C R E A S E I N BODY T E M P E R A T U R E OF THE C H I C K S WAS A L S O R E P O R T E D (P AS TRO _ET _AL . 1969B). THE HORMONAL R E S P O N S E AND CHANGE I N BODY T E M P E R A T U R E WERE S U G G E S T E D BY THE R E S E A R C H E R S AS A B A S I S FOR E X P L A I N I N G D E P R E S S I O N OF FE E D C O N S U M P T I O N I N GROWING A N I M A L S FED I M B A L A N C E D D I E T S . A S I M I L A R H Y P O T H E S I S WAS PROPOSED BY BR O B E C K (1957) THAT A THERMAL R E C E P T O R S I M I L A R TO THAT WHICH MAY O P E R A T E I N T E M P E R A T U R E CONTROL I S A L S O E F F E C T I V E I N R E G U L A T I O N OF F E E D I N G . E A T I N G WAS THOUGHT THE R E S P O N S E TO A F A L L I N HEAT P R O D U C T I O N AND THE S T O P P I N G OF E A T I N G A R E S P O N S E TO A R I S E I N HEAT P R O D U C T I O N . ANDERSON _ET _AL. (1969) R E P O R T E D A R E L A T I O N S H I P BETWEEN THE AMOUNT OF P R O T E I N OR A M I N O A C I D S CONSUMED BY AN A N I M A L AND I T S C A P A C I T Y TO U T I L I Z E AND DEGRADE A S U P P L Y OF A M I N O AC I D S . I T COULD BE THAT THE C A P A C I T Y OF AN A N I M A L FOR A M I N O A C I D D E G R A D A T I O N I N C R E A S E S FROM A LOW TO A H I G H E R V A L U E I N A COLD E N V I R O N M E N T AND C O N S E Q U E N T L Y THE A N I M A L I S A B L E TO EAT MORE OF A D I E T C O N T A I N I N G A S U R P L U S OF A M I N O A C I D S OR P R O T E I N S B E F O R E CHANGES I N THE C O N C E N T R A T I O N AND P A T T E R N OF P L A S M A A M I N O A C I D S BECOME S U F F I C I E N T L Y ABNORMAL TO I N H I B I T FOOD I N T A K E . I F A M I N O A C I D S ARE I N E X C E S S I N THE D I E T , THE C O N C E N T R A T I O N S I N THE T I S S U E S CAN BE REDUCED ONLY BY I N C R E A S E D C A T A B O L I S M , I N C R E A S E D I N C O R P O R A T I O N INTO P R O T E I N OR D E C R E A S E D I N T A K E . I N THE A N I M A L FED A LOW P R O T E I N D I E T , GROWTH RATE I S LOW, P R O T E I N S Y N T H E S I S I S LOW AND A C T I V I T I E S OF E N Z Y M E S OF A M I N O A C I D C A T A B O L I S M ARE LOW (HARPER 1965, 1968), HENCE ANY S U R P L U S OF A M I N O A C I D S CANNOT BE R E A D I L Y OR R A P I D L Y C L E A R E D FROM THE BODY. O R D I N A R I L Y A S U R P L U S OF A M I N O A C I D S WOULD A C C U M U L A T E IN BLOOD ONLY A F T E R THE NEEDS FOR P R O T E I N S Y N T H E S I S HAVE B E E N S A T I S F I E D . E L E V A T E D P L A S M A AM I NO•AC ID C O N C E N T R A T I O N S COULD S E R V E AS A S I G N A L L E A D I N G TO THE C U R T A I L M E N T OF E A T I N G . AMINO ACID I N T E R A C T I O N E X C E S S D I E T A R Y A M I N O A C I D I S T O X I C TO A N I M A L S AND A S P E C I F I C A M I N O A C I D I S R E Q U I R E D TO C O U N T E R A C T THE T O X I C E F F E C T . D'MELLO AND LEWIS (1970) HAVE SHOWN THREE T Y P E S OF S U C H S P E C I F I C I T Y OF I N T E R -A C T I O N BETWEEN A M I N O A C I D S I N C H I C K N U T R I T I O N . THE A D V E R S E - E F F E C T S INDUCED BY E X C E S S L Y S I N E I S A L L E V I A T E D ONLY BY A R G I N I N E , THAT OF L E U C I N E BY V A L I N E OR I S O L E U C I N E AND THAT OF T H R E O N I N E BY T R Y P T O P H A N . E X C E S S D I E T A R Y L Y S I N E I N C R E A S E S A R G I N I N E R E Q U I R E M E N T OF THE C H I C K M A R K E D L Y . H I G H E R A R G I N I N E R E Q U I R E M E N T HAS B E E N I N T E R P R E T E D ( J O N E S , 1964; SMITH AND L E W I S , 1966; 0 'DELL AND S A V A G E , 1966) AS A S P E C I F I C A N T A G O N I S M BETWEEN A R G I N I N E AND L Y S I N E . BOORMAN AND F L S H E R (1966), HOWEVER, I N D I C A T E D THAT THE L Y S I N E - A R G I N I N E I N T E R A C T I O N I S A N O N - S P E C I F I C M A N I F E S T A T I O N OF A G E N E R A L PHENOMENON OF A M I N O A C I D DETOX I CAT I ON. THE L Y S I N E - A R G I N L N E A N T A G O N I S M HAS B E E N S T U D I E D E X T E N S I V E L Y I N THE C H I C K . THREE P O S S I B L E M E C H A N I S M S ARE O F F E R E D TO E X P L A I N THE ROLE OF A R G I N I N E I N S I T U A T I O N S OF A M I N O A C I D I M B A L A N C E : ( A ) E X C E S S A M I N O A C I D S B R I N G ABOUT A R E D U C T I O N I N F E E D I N T A K E R E S U L T I N G I N A D E F I C I E N C Y OF A R G I N I N E FOR P R O T E I N S Y N T H E S I S ; ( B ) A R G I N I N E A C T S AT THE M E T A B O L I C L E V E L BY I N C R E A S I N G THE E F F I C I E N C Y OF E I T H E R OR BOTH THE U R I C A C I D C Y C L E I N T E R M E D I A T E S ( S N E T S I N G E R AND S C O T T , 1961); AND ( C ) A R G I N I N E C O M P E T E S WITH THE A M I N O A C I D I N E X C E S S AT THE A B S O R P T I O N S I T E ( G A S T R O I N T E S T I N A L T R A C T ) SO THAT E X C E S S Q U A N T I T I E S ARE NOT A B S O R B E D AND/OR AT THE R E A B S O R P T I O N S I T E ( K I D N E Y ) SO THAT R E A B S O R P T I ON OF THE E X C R E T E D E X C E S S A M I N O A C I D S DOES NOT OCCUR. THE L A T T E R H Y P O T H E S I S I S BAS E D ON THE O B S E R V A T I O N S OF ROS E N B E R G E T _AL. (1962) WHICH I N D I C A T E A MUTUAL C O M P E T I T I V E I N H I B I T I O N BETWEEN L - A R G I N I N E , L-L Y S I NE AND L - O R N I T H I N E I N THE RAT K I D N E Y . THE T H R E E M E C H A N I S M S WERE R E P U D I A T E D BY SMITH (1968) WHO SHOWED THAT S U P P L E M E N T A L A R G I N I N E I N I M B A L A N C E D D I E T S D I D NOT E X E R T I T S B E N E F I T B E C A U S E OF A D E F I C I E N C Y OF T H I S A M I N O A C I D DUE TO A REDUCED F E E D I N T A K E , D I D NOT M E D I A T E I T S I N F L U E N C E THROUGH C O M P E T I T I V E I N H I B I T I O N AT A B S O R P T I O N OR R E A B S O R P T I O N S I T E S NOR D I D I T A P P E A R TO ACT AS A C Y C L E I N T E R M E D I A T E I N A L A T E N T UREA C Y C L E . F U R T H E R M O R E , A R G I N I N E WOULD NOT BE E X P E C T E D TO F A C I L I T A T E UREA S Y N T H E S I S I N C H I C K S S I N C E B I R D S L A C K A F U N C T I O N I N G UREA C Y C L E ( T A M I R AND R A T N E R , 1963). THERE I S NO KNOWN S T I M U L A T O R Y E F F E C T OF A R G I N I N E ON U R I C A C I D B I O -S Y N T H E S I S AND A T T E M P T S TO R E L I E V E AMMONIA T O X I C I T Y WITH A R G I N I N E I N THE C H I C K HAVE B E E N U N S U C C E S S F U L ( BLOOMF I ELD _ET _AL . 1 969). JONES n A L . (1967) R E P O R T E D THAT L Y S I N E E X C E S S E S C A U S E D I N C R E A S E D K I D N E Y A R G I N A S E A C T I V I T Y , A L T H O U G H HE F E L T THAT THE I N C R E A S E I N A R G I N A S E OCCURRED TOO L A T E TO ACCOUNT FOR THE E L E V A T E D A R G I N I N E R E Q U I R E M E N T . A U S T I C AND N E S H E I M (1970) A L S O O B S E R V E D THAT MARKED I N C R E A S E S I N A R G I N A S E A C T I V I T Y AND UREA E X C R E T I O N WERE A S S O C I A T E D WITH I N C R E A S E S I N THE A R G I N I N E R E Q U I R E M E N T . T l M E-COURSE S T U D I E S , HOWEVER, R E V E A L E D THAT THE I N I T I A L I N C R E A S E S I N K I D N E Y A R G I N A S E A C T I V I T Y AND UREA E X C R E T I O N C O R R E S P O N D E D I N T I M E WITH THE I N I T I A L D E C R E A S E S I N RATE OF GROWTH AND FOOD I N T A K E WHEN C H I C K S WERE FED A R G I N I N E L I M I T I N G D I E T S . THE A L T E R A T I O N OF THE A C T I V I T Y OF T H I S E N Z Y M E WAS PROPOSED AS A MAJOR F A C T O R I N V A R I A T I O N S OF THE A R G I N I N E R E Q U I R E M E N T OF C H I C K S . P O T A S S I U M C A T I O N S U P P L E M E N T A T I O N OF A C A S E I N B A S E D D I E T S T I M U L A T E D C H I C K GROWTH M A R K E D L Y , S P A R E D THE A R G I N I N E R E Q U I R E M E N T AND C R E A T E D MORE F A V O U R A B L E F R E E A M I N O A C I D P A T T E R N S I N THE P L A S M A AND M U S C L E OF C H I C K S ( S T U T Z _ET _AL . 1 9 7 1 ) . B ASED ON T H E S E O B S E R V A T I O N S , A MORE P O S I T I V E C A T I O N B A L A N C E WAS P O S T U L A T E D TO D E C R E A S E THE RATE OF A R G I N I N E C A T A B O L I S M . EXPERIMENTAL PROCEDURE GENERAL E I G H T C H I C K F E E D I N G T R I A L S WERE CONDUCTED TO E X A M I N E THE I M P A C T OF D I E T A R Y L Y S I N E ON THE M E T A B O L I S M OF N U C L E I C A C I D S , P R O T E I N AND A M I N O A C I D . B R O I L E R C H I C K S M A I N T A I N E D I N E L E C T R I C A L L Y H E A T E D B A T T E R Y BROODERS WERE USED AS E X P E R I M E N T A L S U B J E C T S THROUGHOUT THE S T U D Y . D I E T A R Y T R E A T M E N T S WERE ARRANGED I N A RANDOM BLOCK D E S I G N I N E A C H T R I A L . FEED AND WATER WERE S U P P L I E D AD L I B I T U M I N EACH F E E D I N G T R I A L , T E C H N I Q U E S . Q U A N T I T A T I V E S E P A R A T I O N AND D E T E R M I N A T I O N OF DNA, RNA AND P R O T E I N METHODS FOR S E Q U E N T I A L S E P A R A T I O N OF RNA, DNA, L I P I D AND P R O T E I N OF T I S S U E HOMOGENATES S A M P L E S OF C H I C K M U S C L E AND L I V E R STORED AT -20°C WERE A N A L Y Z E D I N D I V I D U A L L Y FOR DNA, RNA AND P R O T E I N BY THE METHOD OF S HI BKO ET A L . (1967) I N T R I A L S 1 TO 3. S A M P L E S OF THE T I S S U E S WERE HOMOGENIZED I N I C E - C O L D S U C R O S E 0.25 M/0.006N NAOH TO A 10$ (w/v) S O L U T I O N , U S I N G A WARING B L E N D E R / O S T E R I Z E R . THE HOMOGENATE WAS S T R A I N E D THROUGH L A Y E R S OF C H E E S E C L O T H TO REMOVE C O N N E C T I V E T I S S U E . ONE ML. OF I C E - C O L D 70$ P E R C H L O R I C A C I D (PCA) WAS ADDED TO A F I V E ML. A L I Q U O T OF HOMOGENATE P L U S F I V E ML. 0.25 M S U C R O S E S O L U T I O N I N A C E N T R I F U G E T U B E . THE S O L U T I O N WAS ALLOWED TO S T A N D FOR 15 M I N U T E S I N AN I C E - B A T H . THE P R E C I P I T A T E FORMED WAS S E P A R A T E D FROM THE S U P E R N A T A N T F L U I D BY C E N T R I F U G A T I ON I N AN I N T E R N A T I O N A L H I G H S P E E D C E N T R I F U G E MODEL HR1 AT 39 4000 RPM AND 4 C. THE S U P E R N A T A N T WAS D I S C A R D E D . THE P R E C I P I T A T E WAS WASHED TWICE WITH 10 ML. P O R T I O N S OF 5$ PCA. E X C E S S A C I D FROM THE L A S T WASH WAS D R A I N E D O F F BY I N V E R T I N G THE C E N T R I F U G E TUBE OVER A P A P E R TOWEL. THE P R E C I P I T A T E WAS THEN S U S P E N D E D I N 18 ML. OF 0.3N N A O H AND I N C U B A T E D FOR ONE HOUR AT 37°C IN A C O N S T A N T - T E M P E R A T U R E W A T E R - B A T H . TWO ML. OF I C E - C O L D 70$ PCA WAS ADDED TO THE R E S U L T I N G N A O H H Y D R O L Y S A T E AND COOLED I N AN I C E - B A T H FOR 10 M I N U T E S . ALL S A M P L E S WERE C E N T R I F U G E D AND THE S U P E R N A T A N T WAS C A R E F U L L Y REMOVED. THE P R E C I P I T A T E WAS WASHED WITH d0 ML. OF 5$ PCA AND C E N T R I F U G E D A G A I N . THE S U P E R N A T A N T S WERE C O M B I N E D FOR RNA D E T E R M I N A T I O N . THE P R E C I P I T A T E WAS S U S P E N D E D I N E I G H T ML. OF 1.5$ PCA AND T H E N I N C U B A T E D FOR 20 M I N U T E S AT 90°C; 0.5 ML. OF 70$ PCA WAS ADDED TO THE S U S P E N S I O N AND T H E N COOLED I N AN I C E - B A T H FOR 10 M I N U T E S . ALL S A M P L E S WERE C E N T R I F U G E D AND THE S U P E R N A T A N T WAS C A R E F U L L Y REMOVED. THE P R E C I P I T A T E WAS WASHED WITH 10 ML. OF 1.5$ PCA AND C E N T R I F U G E D A G A I N , THE S U P E R N A T A N T S WERE C O M B I N E D FOR DNA D E T E R M I N A T I O N . THE P R E C I P I T A T E WAS E X T R A C T E D WITH 20 ML. OF ETHANOL/0.35$ PCA TO REMOVE SOME OF THE L I P I D . A D D I T I O N A L L I P I D E X T R A C T I O N WAS PERFORMED WITH 10 ML. OF A 3:1 E T H A N O L : C H L 0 R O F O R M M I X T U R E , E T H A N O L : E T H E R , I N WHICH THE P R E C I P I T A T E WAS HEATED FOR 15 M I N U T E S AT 37°C AND 5 ML. OF P E T R O L E U M E T H E R THEN ADDED, AND 20 ML. ANHYDROUS E T H Y L E T H E R . THE R E S U L T I N G P E L L E T WAS D R I E D FOR 4 HOURS AT 90°C I N AN O V E N . ALL T I S S U E S A M P L E S WERE RUN I N D U P L I C A T E . D E T E R M I N A T I O N OF RNA, DNA AND P R O T E I N RNA AND DNA C O N C E N T R A T I O N I N T I S S U E E X T R A C T S ( S U P E R N A T A N T S ) WERE 41 D E T E R M I N E D BY U L T R A V I O L E T A B S O R P T I O N M E A S U R E M E N T S AT 260 AND 268.5 MU R E S P E C T I V E L Y I N A UN I C A M SP 1800 S P E C T R O P H O T O M E T E R . P R O T E I N H Y D R O L Y S I S PRODUCTS C O N T A M I N A T I N G RNA AND DNA E X T R A C T S WERE C O R R E C T E D AS FOLLOWS: C O N T A M I N A T I N G P E P T I D E S WERE MEASURED BY THE M O D I F I E D METHOD OF LOWRY _ET _AL. (1951) D E S C R I B E D BY M I L L E R (1959). O P T I C A L D E N S I T I E S (260 MU) OF RNA S O L U T I O N S WERE C O R R E C T E D FOR A B S O R B E N C Y DUE TO P R O T E I N _ 3 BY S U B S T R A C T I N G 1x10 J O P T I C A L D E N S I T Y U N I T PER MICROGRAM P R O T E I N PER M I L L I L I T E R OF S O L U T I O N AS S U G G E S T E D BY MUNRO AND F L E C K (1966). L I K E -_ 3 W I S E , A C O R R E C T I O N F A C T O R OF 0.8X10 O P T I C A L D E N S I T Y U N I T (268.5 MU) PER MICROGRAM P R O T E I N PER M I L L I L I T E R WAS A P P L I E D TO DNA AS I N D I C A T E D BY HOWARTH AND B A L D W I N (1971). P R O T E I N I S O L A T E D FROM, THE HOMOGENATE WAS D R I E D AND WEIGHED. THE P R O T E I N WAS D I S S O L V E D BY H E A T I N G WITH 1N KOH. ONE I'ML. OF THE KOH H Y D R O L Y S A T E WAS D I L U T E D TO 50 ML. WITH D I S T I L L E D WATER. ONE ML. OF THE D I L U T E S O L U T I O N WAS THEN USED TO D E T E R M I N E P R O T E I N C O N T E N T A C C O R D I N G TO THE METHOD D E S C R I B E D BY M I L L E R (1959). THE G R A V I M E T R I C PROCEDURE WAS FOUND TO G I V E V A L U E S I N A G R E E M E N T WITH V A L U E S D E T E R M I N E D BY THE B I U R E T - P H E N O L METHOD OF M I L L E R (1959). STANDARD C U R V E S OF DNA, RNA AND P R O T E I N C O N C E N T R A T I O N S A G A I N S T O P T I C A L D E N S I T Y WERE P R E P A R E D U S I N G C A L F THYMUS DNA, Y E A S T RNA AND B O V I N E S E R U M A L B U M I N R E S P E C T I V E L Y . TOTAL T I S S U E DNA, RNA AND P R O T E I N WERE D E T E R M I N E D FROM THE STANDARD C U R V E S . RNA E X T R A C T S I N T R I A L S 4 AND 5 WERE P R E P A R E D FROM L I V E R AND MU S C L E HOMOGENATES AS I N D I C A T E D BY SHI BKO _ET _AL . (1967). THE E X T R A C T C O N T A I N E D THE 2!, 3'-RI B O N U C L E O T I D E PRODUCTS OF RNA H Y D R O L Y S I S AND I T S A B S O R B A N C E WAS MEASURED AT 232 AND 260 MU I N A U-NICAN SP 1800 42 S P E C T R O P H O T O M E T E R . RNA C O N C E N T R A T I O N WAS C A L C U L A T E D U S I N G A TWO-W A V E L E N G T H FORMULA C-„. = 3.40A„ R„ - 1.44A 0 0 0 D E S C R I B E D FOR THE RNA 260MU 232MU RAT L I V E R RNA BY F L E C K AND BEGG (1954). T H I S FORMULA C O R R E C T E D FOR L I G H T A B S O R B A N C E BY AC I D - S O L U B L E P E P T I D E S I N THE E X T R A C T S AND GAVE S I M I L A R R E S U L T S TO A D I F F E R E N T PROCEDURE USED I N T R I A L S 1-3. P R O T E I N MEASUREMENT WITH THE F O L I N - P H E N O L REAGENT I S O L A T E D T I S S U E P R O T E I N WAS H Y D R O L Y S E D BY H E A T I N G WITH 1N KOH. ONE ML. OF THE KOH HYDROLYSA'TE WAS D I L U T E D TO 50 ML. WITH D I S T I L L E D WATER. ONE ML. OF A L K A L I N E C O P P E R R E A G E N T C O N T A I N I N G 10:1 10$ NA2CO2 I N 0.5N' NA0H:O.5$ CUS0^-1$ S O D I U M T A R T R A T E WAS ADDED TO ONE ML. OF THE P R O T E I N S O L U T I O N . THE M I X T U R E WAS ALLOWED TO S T A N D FOR 10 M I N U T E S AT ROOM T E M P E R A T U R E . THREE ML. OF F O L I N - P H E N O L R E A G E N T ( D I L U T E D 1:11 WITH D I S T I L L E D WATER) WAS ADDED TO THE M I X T U R E U S I N G A HAND P L U N G E R . THE P L U N G E R ALLOWS P R E L I M I N A R Y M I X I N G OF THE P R O T E I N ? S O L U T I O N WITH THE R E A G E N T S AND HENCE WHEN A L A R G E NUMBER OF S A M P L E S ARE B E I N G A N A L Y Z E D , E A C H T E S T TUBE C O N T A I N I N G THE P R O T E I N S O L U T I O N AND R E A G E N T S NEED NOT BE S H A K E N U N T I L A F T E R THE R E A G E N T S HAVE B E E N ADDED TO A L L S A M P L E S . THE S A M P L E S WERE H E A T E D FOR 10 M I N U T E S AT 50°C I N A C O N S T A N T T E M P E R A T U R E WATER-BATH AND THEN ALLOWED TO COOL TO ROOM T E M P E R A T U R E . L I G H T A B S O R P T I O N M E A S U R E M E N T S OF THE P R O T E I N - C U - P H E N O L C O M P L E X FORMED WERE MADE AT 750 MU I N A BECKMAN S P E C T R O P H O T O M E T E R . T I S S U E P R O T E I N C O N T E N T WAS THEN C A L C U L A T E D FROM THE STANDARD B O V I N E S E R U M A L B U M I N C U R V E . E X P E R I M E N T A L DATA WERE A N A L Y Z E D BY A N A L Y S I S OF V A R I A N C E , SNEDECOR (1956). THE T R E A T M E N T D I F F E R E N C E S WERE T E S T E D BY THE M U L T I P L E RANGE T E S T OF DUNCAN (1955). TRIAL 1. DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N T I S S U E . ONE HUNDRED AND E I G H T Y B R O I L E R C H I C K S (ONE DAY OF A G E ) WERE RANDOMLY D I S T R I B U T E D INTO 15 GROUPS OF 12 C H I C K S E A C H . THE F I V E D I E T S P R O V I D E D I N T H I S T R I A L C O N S I S T E D OF A CONTROL W H E A T - B A S A L C O N T A I N I N G 0.36$ L Y S I N E (TABLE 1), AND FOUR OTHER D I E T S F O R M U L A T E D BY A D D I N G 0.25, 0.50, 0.75 AND 1.0$ L - L Y S I N E TO T H I S CONTROL D I E T . EACH D I E T WAS FED TO TH R E E R E P L I C A T E GROUPS OF C H I C K S . D A I L Y F E E D I N T A K E AND WE E K L Y I N D I V I D U A L BODY WEIGHT WERE RECORDED. Six C H I C K S FROM E A C H D I E T A R Y GROUP WERE S A C R I F I C E D ON THE 14TH DAY AND T H E I R L I V E R S AND G A S T R O C N E M I U S M U S C L E REMOVED. THE L I V E R S AND M U S C L E S WERE WEIGHED AND STORED I N P L A S T I C BAGS AT -20°C U N T I L A N A L Y Z E D I N D I V I D U A L L Y FOR DNA, RNA AND P R O T E I N . C H I C K S C O N S U M I N G THE WHEAT B A S A L D I E T , AND THE B A S A L P L U S 0.25, 0.50 AND 0.75$ L Y S I N E WERE C O N T I N U E D ON THE D I E T S FOR 28 D A Y S . ON THE L A S T DAY OF THE T R I A L , S I X C H I C K S FROM E A C H D I E T A R Y GROUP WERE S A C R I F I C E D . THE L I V E R S AND M U S C L E S WERE WEIGHED AND STORED U N T I L THE DNA, RNA AND P R O T E I N C O N T E N T WERE D E T E R M I N E D . TRIAL 2. DNA, RNA AND P R O T E I N C O N C E N T R A T I O N I N T I S S U E . A S E C O N D T R I A L WAS RUN TO F U R T H E R E X A M I N E THE H Y P O T H E S I S THAT E X C E S S L Y S I N E WOULD D E P R E S S TOTAL L I V E R AND MUS C L E DNA, RNA AND P R O T E I N A C C U M U L A T I O N . THE P O S S I B I L I T Y OF A R G I N I N E A L L E V I A T I N G THE I L L - E F F E C T S OF E X C E S S L Y S I N E ON THE A C C U M U L A T I O N OF T I S S U E C O N S T I T U E N T S AS I T DOES ON C H I C K BODY.GROWTH WAS A L S O E X A M I N E D . ONE HUNDRED AND E I G H T C H I C K S (oNE DAY OF A G E ) WERE RANDOMLY TABLE 1 . - - C O M P O S i T i O N OF WHEAT B A S A L D I E T I N G R E O I E N T P E R C E N T A G E OF DIE T WHEAT (16$ P R O T E I N ) 95. 0 V E G E T A B L E O I L 1. 0 T R I C A L C I U M P H O S P H A T E 2. 0 L 1 M E S T O N E 1 V I T A M I N P R E M I X 1. 0 0. 5 MINERAL P R E M I X 0. 5 P R O T E I N ( C A L C U L A T E D V A L U E , $ ) 15. 2 M E T A B O L 1 Z A B L E E N E R G Y ( C A L C U L A T E D V A L U E , K C A L / K G . ) 2930 V I T A M I N P R E M I X S U P P L I E D THE F O L L O W I N G AMOUNT PER KG. OF D I E T : V I T A M I N A, 11000 I.U.; V I T A M I N 0^, 825 I.C.U.; V I T A M I N E, 18.2 I.U.; V I T A M I N K, 1.1 MG.; N I A C I N , 35.2 MG.; R I B O F L A V I N , 5.0 MG.; P A N T O T H E N I C A C I D , 24.2 MG.; P Y R I D O X I N E HC1, 2.86 MG.; C H O L I N E C H L O R I D E , 2.6 G.; B I O T I N , 90.0 MCG.; V I T A M I N B.^, 13.2 MCG.; F O L I C A C I D , 550.0 MCG. I MINERAL P R E M I X S U P P L I E D THE F O L L O W I N G AMOUNT PER KG. OF D I E T : M A N G A N E S E , 86.0 MG.; Z I N C , 49.0 MG.; C O P P E R , 7.7 MG.; I R O N , 34.0 MG. AND S O D I U M C H L O R I D E ( I O D I Z E D ) , 2.75 G. 'HUBBELL, C. H., 1969. A N A L Y S I S T A B L E FOR F E E D I N G R E D I E N T S . F E E D S T U F F S , M I L L E R P U B L I S H I N G CO., M I N N E A P O L i s . 45 D I S T R I B U T E D INTO N I N E GROUPS OF 12 C H I C K S E A C H . T R I P L I C A T E LOTS OF C H I C K S WERE FED EACH OF THE F O L L O W I N G D I E T S : THE W H E A T - B A S A L D I E T (TABLE 1), AND THE W H E A T - B A S A L D I E T S U P P L E M E N T E D WITH 0.5, 1.0 AND 1.5$ L - L Y S I N E . BODY WEIGHT AND F E E D I N T A K E WERE R E S P E C T I V E L Y RECORDED WEEKLY AND D A I L Y . Six B R O I L E R C H I C K S PER D I E T A R Y T R E A T M E N T WERE S A C R I F I C E D ON DAY 14. L I V E R S AND G A S T R O C N E M I U S M U S C L E WERE REMOVED, WEIGHED AND F R O Z E N (-20°C) U N T I L A N A L Y Z E D FOR DNA, RNA AND P R O T E I N C O N T E N T . S T A R T I N G FROM DAY 14, THE WHEAT B A S A L P L U S 1.0$ L - L Y S I N E D I E T WAS S U P P L E M E N T E D WITH 0.013$ L - A R G I N I N E WHILE THE WHEAT B A S A L P L U S 1.5$ L - L Y S I N E D I E T WAS S U P P L E M E N T E D WITH 0.293$ L - A R G I N I N E . THE F E E D I N G WAS C O N T I N U E D FOR ANOTHER 14 D A Y S . BODY WEIGHT AND F E E D I N T A K E WERE RECORDED W E E K L Y . Six C H I C K S PER D I E T A R Y T R E A T M E N T WERE S A C R I F I C E D AT THE END OF THE T R I A L . L l V E R AND G A S T R O C N E M I U S M U S C L E S WERE REMOVED, WEIGHED AND F R O Z E N U N T I L A N A L Y Z E D FOR DNA, RNA AND P R O T E I N . TRIAL 3. DNA,,1, RNA AND P R O T E I N C O N C E N T R A T I O N I N T I S S U E . IN THE TWO P R E V I O U S T R I A L S , THE I M P A C T OF D I E T A R Y L Y S I N E ON T I S S U E DNA, RNA AND P R O T E I N S Y N T H E S I S AND A C C U M U L A T I O N WAS M E A S U R E D . THIS T R I A L WAS CONDUCTED TO S T U D Y THE R E L A T I V E E F F E C T S OF THE Q U A N T I T Y AND Q U A L I T Y OF D I E T A R Y P R O T E I N ON THE S Y N T H E S I S AND A C C U M U L A T I O N OF DNA, RNA AND P R O T E I N I N THE L I V E R AND G A S T R O C N E M I U S M U S C L E . SEVENTY-TWO B R O I L E R C H I C K S (ONE DAY OF A G E ) , HUBBARD S T R A I N , WERE RANDOMLY D I V I D E D INTO S I X GROUPS OF 12 C H I C K S E A C H . T H R E E GROUPS WERE FED THE W H E A T - S O Y B E A N (15$ P R O T E I N ) D I E T AND THE OTHER T H R E E GROUPS R E C E I V E D THE W H E A T - S 0 Y B E A N - F I S H M E A L (20$ P R O T E I N ) D I E T P R E S E N T E D I N T A B L E 2. BOD Y WEIGHT AND F E E D I N T A K E WERE RECORDED W E E K L Y . S I X C H I C K S P E R D I E T A R Y T R E A T M E N T WERE S A C R I F I C E D ON DAY 14 AND ON DAY 28 R E S P E C T I V E L Y . THE L I V E R S AND G A S T R O C N E M I U S M U S C L E S WERE REMOVED, WEIGHED AND F R O Z E N U N T I L A N A L Y Z E D FOR DNA, RNA AND P R O T E I N , THE R E S U L T I N G DATA WERE COMPARED WITH THOSE O B T A I N E D FROM F E E D I N G THE W H E A T - B A S A L D I E T AND THE W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E D I E T I N T R I A L 1. THE C O M P A R I S O N I S V A L I D S I N C E BOTH T R I A L S WERE RUN S I M U L T A N E O U S L Y . TRIAL 4. TURNOVER RATE OF T I S S U E P R O T E I N . THE I N F L U E N C E OF D I E T A R Y L Y S I N E ON THE RATE OF P R O T E I N S Y N T H E S I S , A C C U M U L A T I O N AND P R O T E I N D E G R A D A T I O N I N THE L I V E R AND T I B I A M I X E D M U S C L E S WAS S T U D I E D I N MORE D E T A I L I N T H I S T R I A L . T H I R T Y - S I X B R O I L E R C H I C K S (ONE DAY OF A G E ) WERE RANDOMLY D I V I D E D INTO THREE GROUPS OF 12 C H I C K S E A C H . THE WHEAT B A S A L D I E T (TABLE 1) WAS FED TO ONE GROUP; A SECOND GROUP R E C E I V E D W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E AND THE T H I R D GROUP R E C E I V E D W H E A T - B A S A L P L U S 0.875$ L - L Y S I N E FOR 15 D A Y S . ON THE S E V E N T H DAY OF GROWTH, EACH C H I C K FROM THE THREE D I E T A R Y T R E A T M E N T S WAS I N J E C T E D I N T R A P E R I T O N E A L L Y WITH 1.0 UC I OF L - L Y S I N E - U - ^ C MONOH YD ROC HLOR I D E (342MC|/M M O L E ) I N 1.0 ML. OF 0.9$ S A L I N E S O L U T I O N . THREE C H I C K S FROM E A C H D I E T A R Y T R E A T M E N T WERE S A C R I F I C E D ON DAY 1, 2, 4 AND 8 F O L L O W I N G THE A D M I N I S T R A T I O N OF R A D I O I S O T O P E . L I V E R S TABLE 2.--COMPOSiTiON OF E X P E R I M E N T A L D I E T S DIET  3 4 I N G R E D I E N T S $ fo WHEAT (13$ P R O T E I N ) 85.5 72.5 SOYBEAN MEAL (48.5$) 9.0 17.5 F I S H MEAL (70$) 3.0 V E G E T A B L E O I L 1 .5 3.0 T R I C A L C I U M P H O S P H A T E . 2.0 2.0 LI M ESTONE V I T A M I N P R E M I X ^ 1 .0 1 .0 0.5 0.5 MINERAL P R E M I X ^ 0.5 0.5 1 AND 2 S U P P L I E D I N THE SAME AMOUNT AS I N T A B L E 1. 48 A N D T I B I A M I X E D M U S C L E S WERE R E M O V E D , W E I G H E D A N D F R O Z E N U N T I L A N A L Y Z E D F O R RNA, P R O T E I N A N D R A D I O A C T I V I T Y I N T I S S U E P R O T E I N . M E A S U R E M E N T S O F R A D I O A C T I V I T Y I N T I S S U E P R O T E I N . P R O T E I N WAS Q U A N T I T A T I V E L Y S E P A R A T E D F R O M T H E L I V E R OR M U S C L E H O M O G E N A T E U S E D I N RNA D E T E R M I N A T I O N (SH I B K O _ET JA_L . 1967). THE P E L L E T L E F T B E H I N D A F T E R T H E E X T R A C T I O N O F RNA WAS T R E A T E D T W I C E W I T H A N E T H A N O L - E T H E R M I X T U R E (3:1) A T 55°C F O R 30 M I N U T E S A N D W A S H E D T W I C E W I T H WARM E T H E R ( Y A M A S H I T A A N D A S H I D A , 1969). T H E R E S U L T A N T P R E C I P I T A T E WAS D I S S O L V E D B Y H E A T I N G W I T H I N KOH. T H E N 1 M L . O F H Y D R O X I D E O F HYAM I N E 10-X (1 M I N M E T H A N O L ) WAS A D D E D TO O N E M L . O F E A C H S O L U T I O N A N D T H E S A M P L E WAS S H A K E N U N T I L C L E A R . TO T H I S C L E A R H Y A M I N E S O L U T I O N , 10.0 M L . O F P - D I O X A N E S C I N T I L L A T I O N M E O I U M WAS A D D E D , A N D T H E S A M P L E WAS C O U N T E D I N A N I S O C A P / 3 0 0 L I Q U I D S C I N T I L L A T I O N S P E C T R O M E T E R A T 4°C. I N T E R N A L S T A N D A R D S WERE U S E D TO E V A L U A T E Q U E N C H I N G , A N D T H E C O R R E C T E D R A D I O A C T I V I T Y V A L U E S E X P R E S S E D A S D I S I N T E G R A T I O N S / M I N U T E ( D P M ) . THE P - D I O X A N E S C I N T I L L A T I O N M E D I U M C O N S I S T E D O F 1OG. O F 2 , 5 - D I P H E N Y L O X A Z O L E (PPO), 0.250 G. O F 1, 4 B I S ( 4 - M E T H Y L - 5 - P H E N Y L O X A Z O L Y L ) B E N Z E N E ( D I M E T H Y L POPOP), A N D 100 G. N A P H T H A L E N E D I S S O L V E D I N O N E L I T E R O F P - D I O X A N E ( W A N N E M A C H E R , J R . _ET _ A L . 1965). ONE M L . O F T H E KOH H Y D R O L Y S A T E WAS D I L U T E D TO 50 M L . W I T H D I S T I L L E D W A T E R . ONE M L . O F T H E D I L U T E S O L U T I O N WAS T H E N U S E D TO D E T E R M I N E P R O T E I N C O N T E N T (M lLLER, 1959). TRIAL 5. S Y N T H E S I S O F T I S S U E RNA. THE I N F L U E N C E O F D I E T A R Y L Y S I N E ON RNA S Y N T H E S I S I N T H E L I V E R A N D M U S C L E WAS S T U D I E D . B R O I L E R C H I C K S ( O N E D A Y O F A G E ) W E R E D I V I D E D 49 I N T O T H R E E G R O U P S . ONE G R O U P WAS F E D T H E W H E A T - B A S A L D I E T ( C O N T A I N I N G 0.36$ L Y S I N E ) . A S E C O N D G R O U P R E C E I V E D T H E W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E A N D T H E T H I R D G R O U P WAS F E D W H E A T - B A S A L P L U S 0.875$ L - L Y S I N E . D I E T S WERE F E D TO E X P E R I M E N T A L C H I C K S D U R I N G A P E R I O D O F E I G H T D A Y S . T H R E E C H I C K S F R O M E A C H D I E T A R Y T R E A T M E N T W E R E E A C H I N J E C T E D 1 4 I N T R A P E R I T O N E A L L Y W I T H 1.0 UCI O F U R A C I L - 2 - C (60 MC I/M M O L E ) I N O N E M L . O F 0.9$ S A L I N E S O L U T I O N ON T H E S E V E N T H D A Y O F T H E G R O W T H T E S T . C H I C K S WERE S A C R I F I C E D 24 H O U R S A F T E R T H E A D M I N I S T R A T I O N O F R A D I O -I S O T O P E . L I V E R S A N D T I B I A M I X E D M U S C L E S WERE R E M O V E D , W E I G H E D A N D F R O Z E N U N T I L A N A L Y Z E D F O R RNA A N D R A D I O A C T I V I T Y . M E A S U R E M E N T S O F R A D I O A C T I V I T Y I N T I S S U E RNA. THE L I V E R A N D M U S C L E W E R E P R E P A R E D F O R A N A L Y S I S A C C O R D I N G TO T H E M E T H O D O F SH I B K 0 _ET _AL . (1967). THE R E S U L T I N G P R O D U C T S O F RNA H Y D R O L Y S I S I N 0.3N NAOH W E R E U S E D F O R RNA C O N C E N T R A T I O N A N D R A D I O -A C T I V I T Y D E T E R M I N A T I O N S . A L I Q U O T S O F T H E S O L U T I O N O F T H E RNA H Y D R O L Y S I S P R O D U C T S I N NAOH W E R E U S E D TO D E T E R M I N E T H E C O N C E N T R A T I O N O F RNA B Y U L T R A V I O L E T A B S O R P T I O N M E A S U R E M E N T S . R A D I O A C T I V I T Y I N T I S S U E H O M O G E N A T E WAS E S T I M A T E D B Y A D D I N G 1 M L . O F H Y D R O X I D E O F H Y A M I N E 10-X TO 1 M L . O F T H E NAOH H Y D R O L Y S A T E A N D T H E S A M P L E WAS S H A K E N U N T I L C L E A R . TO T H I S C L E A R H Y A M I N E S O L U T I O N , 10.0 M L . O F P - D I O X A N E S C I N T I L L A T I O N M E D I U M ( W A N N E M A C H E R , J R . E T A L . 1965) WAS A D D E D , A N D T H E S A M P L E WAS C O U N T E D A S I N T R I A L 4. RNA R A D I O A C T I V I T Y WAS E S T I M A T E D B Y P U T T I N G 1.0 M L . O F T H E RNA E X T R A C T I N S C I N T I L L A T I O N G L A S S V I A L S . T H E RNA E X T R A C T S W E R E N E U T R A -L I Z E D W I T H 6N KOH, 10.0 M L . O F P - D I O X A N E S C I N T I L L A T I O N M E D I U M WAS A D D E D A N D T H E S A M P L E WAS C O U N T E D . TRIAL 6. M E T A B O L I C F A T E O F A M I N O A C I D S . THE I N F L U E N C E O F D I E T A R Y L Y S I N E , T H R E O N I N E A N D A R G I N I N E ON T H E 14 14 O X I D A T I O N O F C F R O M L - L Y S I N E - U - C TO R A D I O A C T I V E C A R B O N D I O X I D E WAS I N V E S T I G A T E D . B R O I L E R C H I C K S ( O N E D A Y O F A G E ) W E R E F E D T H E W H E A T B A S A L P L U S 0.5$ L-L Y S I N E D I E T F O R S E V E N D A Y S . TWO H U N D R E D A N D F I F T Y -TWO C H I C K S W E R E T H E N R A N D O M L Y D I V I D E D I N T O 21 G R O U P S O F 12 C H I C K S E A C H . T H R E E G R O U P S O F C H I C K S W E R E C O N T I N U E D ON T H E W H E A T - B A S A L P L U S 0.5$ L-L Y S 1 N E D I E T F O R A N O T H E R F I V E W E E K S . T H E R E M A I N I N G 18 G R O U P S ( T H R E E G R O U P S P E R T R E A T M E N T ) O F C H I C K S W E R E F E D F O R F I V E W E E K S T H E F O L L O W I N G D I E T S : (1) W H E A T - B A S A L P L U S 0.875$ L - L Y S I N E ; (2) W H E A T -B A S A L P L U S 0.875$ L - L Y S I N E A N D 0.4$ L - A R G I N I N E ; (3) W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E A N D 0.16$ L~ T H R E O N I N E (4) WH E A T- S 0 Y B E A N (15$ P R O T E I N , T A B L E 2); (5) W H E A T - S O Y B E A N P L U S 0.375$ L - L Y S I N E A N D (6) W H E A T -S O Y B E A N P L U S 0.375$ L - L Y S I N E A N D 0.4$ L - A R G I N I N E . B O D Y W E I G H T A N D F E E D C O N S U M P T I O N WERE R E C O R D E D W E E K L Y . D U R I N G T H E L A S T F I V E D A Y S O F T H E G R O W T H S T U D Y , T H R E E C H I C K S F R O M E A C H D I E T A R Y T R E A T M E N T WERE U S E D TO S T U D Y L Y S I N E O X I D A T I O N . E A C H 1 4 C H I C K WAS I N J E C T E D I N T R A P E R I T O N E A L L Y W I T H 1.0 uCI O F L - L Y S I N E - U - C M O N O H Y D R O C H L O R I DE (342MCI/M M O L E ) I N 2.0 M L . O F 0.9$ S A L I N E S O L U T I O N . I M M E D I A T E L Y F O L L O W I N G I N J E C T I O N , E A C H C H I C K WAS P L A C E D I N A R E S P I R A T I O N C H A M B E R D U R I N G A O N E H O U R C O L L E C T I O N P E R I O D . E X P I R E D G A S E S W E R E P U L L E D T H R O U G H TWO C A R B O N D I O X I D E T R A P S , C O N S I S T I N G O F 20 M L . O F A 1:7 M I X T U R E O F E T H A N O L A M I N E - E T H Y L E N E G L Y C O L M O N O M E T H Y L E T H E R ( j E F F A Y A N D A L V A R E Z , 1961). TO 8 M L . O F T R A P P I N G S O L U T I O N , 2 M L . O F E T H Y L E N E G L Y C O L M O N O M E T H Y L E T H E R WERE A D D E D TO P R E V E N T T H E P R E -C I P I T A T I O N O F E T H A N O L A M I N E C A R B O N A T E , F O L L O W E D B Y 10 M L . O F S C I N T I L L A T I O N F L U I D C O N T A I N I N G 5.0 G. 2 , 5 - D I P H E N Y L O X A Z O L E P E R L I T E R A N D T H E S O L U T I O N WAS T H E N C O U N T E D . TRIAL 7. M E T A B O L I C F A T E O F A M I N O A C I D S . THE I N F L U E N C E O F D I E T A R Y L Y S I N E ON T H E M E T A B O L I C F A T E O F L Y S I N E A N D T H R E O N I N E WAS S T U D I E D . TWO-WEEK O L D C H I C K S F E D A C O M M E R C I A L B R O I L E R D I E T W E R E U S E D . E I G H T E E N C H I C K S W E R E R A N D O M L Y D I V I D E D I N T O T H R E E G R O U P S O F S I X C H I C K S E A C H . ONE G R O U P WAS F E D T H E W H E A T - B A S A L D I E T (TABLE 1); A N O T H E R G R O U P R E C E I V E D T H E W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E A N D T H E T H I R D G R O U P R E C E I V E D W H E A T - B A S A L P L U S 0.875$ L - L Y S I N E . ON T H E S I X T H D A Y O F F E E D I N G , TWO C H I C K S F R O M E A C H D I E T A R Y T R E A T M E N T W E R E U S E D TO E X A M I N E L Y S I N E M E T A B O L I S M ; A N D T H E R E M A I N I N G F O U R C H I C K S I N E A C H D I E T A R Y G R O U P W E R E U S E D TO E X A M I N E T H R E O N I N E M E T A B O L I S M . E A C H C H I C K WAS I N J E C T E D I N T R A P E R I T O N E A L L Y W I T H 0.5 UCI O F L - L Y S I N E - U - ^ C M O N O H Y D R O C H L O R I D E (342MC|/M M O L E ) I N 1.0 M L . O F 0.9$ S A L I N E S O L U T I O N I N T H E G R O U P U S E D T O E X A M I N E L Y S I N E M E T A B O L I S M ; 14 A N D W I T H 1.0 UC I O F L - T H R E ON I N E - U - C (208MCI/M M O L E ) I N 2.0 M L . O F 0.9$ S A L I N E S O L U T I O N I N T H E G R O U P U S E D TO E X A M I N E T H R E O N I N E M E T A B O L I S M . F O L L O W I N G T H E I N J E C T I O N , C H I C K S WERE R E T U R N E D TO T H E R E S P E C T I V E D I E T S F O R A N O T H E R TWO H O U R S . TWO H O U R S A F T E R T H E I N J E C T I O N O F R A D I O I S O T O P E , T H E C H I C K S W E R E S A C R I F I C E D . THE L I V E R S A N D T I B I A M I X E D M U S C L E S W E R E R E M O V E D , W E I G H E D A N D F R O Z E N U N T I L A N A L Y Z E D F O R P R O T E I N A N D R A D I O A C T I V I T Y . M E A S U R E M E N T S O F R A D I O A C T I V I T Y I N T I S S U E P R O T E I N S . M E A S U R E M E N T S O F R A D I O A C T I V I T Y I N A C I D - S O L U B L E P R O T E I N F R A C T I O N A N D P R O T E I N I N T H E L I V E R A N D M U S C L E W E R E M A D E W I T H A M O D I F I E D P R O C E D U R E O F Y A M A S H I T A A N D A S H I D A (1969). A P O R T I O N O F T H E L I V E R OR M U S C L E WAS H O M O G E N I Z E D I N A W A R I N G B L E N D E R U S I N G 9 V O L U M E S O F I C E - C O L D 5.6$ T R I C H L O R O A C E T I C A C I D (TCA) TO F R A C T I O N A T E T H E A C I D - S O L U B L E F R A C T I O N A N D P R O T E I N . ill H E T O T A L R A D I O A C T I V I T Y I N T H E A C I D - S O L U B L E F R A C T I O N WAS D E T E R M I N E D A F T E R D R Y I N G 0.3 M L . O F T H E S U P E R N A T A N T F R A C T I O N I N A C O U N T I N G V I A L , F O L L O W E D B Y T H E A D D I T I O N O F 0.3 M L . H Y A M I N E 1 OX A N D 10 M L . P H O S P H O R S O L U T I O N (4G. 2 , 5 - D I P H E N Y L O X A Z O L E (PPO) A N D 0.1G. 1,4 B L S ( 2 - ( 5 P H E N Y L O X A Z O L Y L ) ) - B E N Z E N E (POPOP) I N 1000 M L . T O L U E N E ) . THE P R E C I P I T A B L E F R A C T I O N O F T H E T C A - E X T R A C T E D T I S S U E S WAS W A S H E D T W I C E W I T H 5$ TCA, H E A T E D A T 90°C F O R 15 M I N U T E S I N T H E S A M E S O L U T I O N , A N D W A S H E D A G A I N . THEN I T WAS T R E A T E D T W I C E W I T H A N E T H A N O L - E T H E R M I X T U R E (3:1) A T 55°C F O R 30 M I N U T E S , A N D F I N A L L Y W A S H E D T W I C E W I T H WARM E T H E R . THE R E S U L T A N T P R E C I P I T A T E WAS D I S S O L V E D B Y H E A T I N G W I T H 1N KOH. THEN 1 M L . O F H Y D R O X I D E O F H Y A M I N E 10-X WAS A D D E D TO 1 M L . O F E A C H S O L U T I O N I N A C O U N T I N G V I A L . T H I S S O L U T I O N WAS S H A K E N U N T I L C L E A R A N D T H E N M I X E D W I T H A 10.0 M L . O F P - D I O X A N E S C I N T I L L A T I O N M E D I U M ( W A N N E M A C H E R _ET _ A L . 1965) A N D C O U N T E D . TRIAL 8. M E T A B O L I C F A T E O F A M I N O A C I D S . THE I N F L U E N C E O F D I E T A R Y L Y S I N E U P O N T H E M E T A B O L I S M O F L Y S I N E A N D T H R E O N I N E WAS S T U D I E D F U R T H E R I N A G R E A T E R D E T A I L . T H R E E H U N D R E D A N D T H I R T Y C H I C K S ( O N E D A Y O F A G E ) W E R E R A N D O M L Y D I V I D E D I N T O 30 G R O U P S O F 11 E A C H . T H R E E P E N S W E R E F E D F O R F O U R W E E K S E A C H O F T H E F O L L O W I N G D I E T S : (1) T H E W H E A T - B A S A L D I E T ( T A B L E 1); (2) W H E A T - B A S A L P L U S 0.084$ L - T H R E O N I N E ; (3) W H E A T - B A S A L P L U S 0.168$ L - T H R E O N I N E ; (4) W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E ; (5) W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E A N D 0.084$ [ . - T H R E O N I N E ; (6) W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E A N D 0.168$ L - T H R E O N I N E ; (7) W H E A T - B A S A L P L U S 0.875$ L - L Y S I N E ; (8) W H E A T - B A S A L P L U S 0.875$ L - L Y S I N E A N D 0.084$ L - T H R E O N I N E ; (9) W H E A T - B A S A L P L U S 0.875$ L - L Y S I N E A N D 0.168$ L - T H R E O N I N E A N D (10) W H E A T - B A S A L P L U S 0.5$ L - L Y S I N E , 0.168$ L - T H R E O N I N E A N D 0.15$ D L - M E T H I ON I N E . W E E K L Y B O D Y W E I G H T A N D F E E D I N T A K E W E R E R E C O R D E D . D U R I N G T H E L A S T S E V E N D A Y S O F T H E GROWTH S T U D Y , T H R E E C H I C K S F R O M E A C H O F D I E T S 1 TO 9 W E R E U S E D TO E X A M I N E L Y S I N E M E T A B O L I S M . T H R E E C H I C K S F R O M E A C H O F D I E T S 1 TO 9 WERE A L S O U S E D TO E X A M I N E T H R E O N I N E M E T A B O L I S M . E A C H C H I C K U S E D TO E X A M I N E L Y S I N E M E T A B O L I S M 1 4 WAS I N J E C T E D I N T R A P E R I T O N E A L L Y W I T H 1.0 uCI O F L - L Y S I N E - U - C M O N O H Y D R O C H L O R I D E (342 MC|/M M O L E ) I N 0.5 M L . O F 0.9$ S A L I N E S O L U T I O N . C H I C K S U S E D TO E X A M I N E T H R E O N I N E M E T A B O L I S M W E R E I N J E C T E D I N T R A -P E RI T O N E A L L Y W I T H 1.0 uCI O F L - T H R E O N I N E - U - ^ ( 2 0 8 MCI/M M O L E ) I N 0.5 M L . O F 0.9$ S A L I N E S O L U T I O N . I M M E D I A T E L Y F O L L O W I N G I N J E C T I O N , E A C H C H I C K WAS P L A C E D I N A R E S P I R A T I O N C H A M B E R . E X P I R E D G A S E S W E R E P U L L E D T H R O U G H TWO C A R B O N D I O X I D E T R A P S F O R TWO H O U R S , A F T E R W H I C H T H E C H I C K WAS S A C R I F I C E D A N D T H E L I V E R A N D T I B I A M U S C L E S W E R E R E M O V E D , W E I G H E D A N D F R O Z E N . T H E M E A S U R E M E N T S O F R A D I O A C T I V I T Y I N T H E R E S P I R A T O R Y CC^, L I V E R A N D M U S C L E P R O T E I N A N D A C I D - S O L U B L E F R A C T I O N W E R E C A R R I E D O U T F O L L O W I N G T H E M E T H O D D E S C R I B E D I N T R I A L S 6 A N D 7. A N A L Y S I S O F V A R I A N C E O F D A T A C O L L E C T E D I N T H E P R E S E N T T R I A L I S I L L U S T R A T E D I N T H E A P P E N D I X T A B L E S . S T A N D A R D C U R V E S O F DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N S A G A I N S T O P T I C A L D E N S I T Y U S I N G C A L F T H Y M U S DNA, Y E A S T RNA A N D B O V I N E S E R U M A L B U M I N A R E P R E S E N T E D I N T H E A P P E N D I X . S T A N D A R D G R A P H S U S E D TO C O R R E C T F O R L O S S O F C O U N T I N G E F F I C I E N C Y I N T H E R A D I O A C T I V E E X P E R I M E N T S A R E A L S O P R E S E N T E D I N T H E A P P E N D I X . RESULTS AND DISCUSSION TRIAL I-T H E E F F E C T S O F D I E T A R Y L Y S I N E ON DNA, RNA, P R O T E I N C O N T E N T A N D W E I G H T O F .THE G A S T R O C N E M I U S M U S C L E F R O M C H I C K S A T TWO W E E K S O F A G E A R E P R E S E N T E D I N T A B L E 3. THE E F F E C T O F D I E T A R Y T R E A T M E N T ON L I V E R W E I G H T , DNA, RNA, A N D P R O T E I N C O M P O S I T I O N A R E I L L U S T R A T E D I N T A B L E S 4 A N D 5. G A S T R O C N E M I U S M U S C L E A N D L I V E R W E I G H T S R E S P O N D E D M A R K E D L Y TO D I E T A R Y L Y S I N E T R E A T M E N T . I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36$ ( W H E A T - B A S A L D I E T ) TO 0.61$ W I T H L - L Y S I N E C A U S E D A S I G N I F I C A N T (P < 0.05) I N C R E A S E I N W E I G H T O F B O T H O R G A N S O V E R T H O S E I N T H E W H E A T - B A S A L D I E T . A P R O G R E S S I V E B U T N 0 N - S I G N I F I C A N T I N C R E A S E I N B O T H M U S C L E A N D L I V E R W E I G H T S R E S U L T E D B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.61 TO 0.86 A N D 1.11$. H O W E V E R , A H I G H E R L E V E L O F D I E T A R Y L Y S I N E (1.36$) D E P R E S S E D M U S C L E W E I G H T S I G N I F I C A N T L Y (P < 0.05). M U S C L E W E I G H T W I T H 1.36$ D I E T A R Y L Y S I N E WAS S I M I L A R TO T H A T O B T A I N E D W I T H T H E 0.36$ L Y S I N E D I E T . L L V E R W E I G H T A T 1.36$ L Y S I N E WAS C O M P A R A B L E TO T H E W E I G H T O B S E R V E D W I T H 0.86 A N D 1.11$ D I E T A R Y L Y S I N E . L L V E R W E I G H T E X P R E S S E D A S A P E R C E N T A G E O F B O D Y W E I G H T A L S O I M P R O V E D M A R K E D L Y (P <0.01) B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36 TO 0.61$. NO S I G N I F I C A N T I M P R O V E M E N T O C C U R R E D A T L E V E L S O F D I E T A R Y L Y S I N E B E Y O N D 0.61$. T H E I N C R E A S E I N G A S T R O C N E M I U S M U S C L E A N D L I V E R W E I G H T S W I T H 55 TABLE 3 . — G A S T R O C N E M I U S M U S C L E W E I G H T , DNA, RNA A N D P R O T E I N C O N T E N T F R O M C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E F O R TWO W E E K S * L Y S i N E T R E A T M E N T S U P P . L E V E L M U S C L E ^ WT. MG . ,DNA MG . MG . ,RNA MG. MG , MG 7^ P R O T E I N M G. 0.25 0.50 0.75 1 .00 0.36 0.61 0.86 1.11 1 .36 681 1162.4 1271 1373.0 632 0.52 0.5o' 0.54' 0.55' 0.55' 0.36. 0.58 0.68E 0.74E 0.34' B 1 .38 1 .57' 1 .79' 1 .67' 1 .68' 0.94 1 .82E 2.30 2.30E 1 .06* B 103.7 129.3' 143.9' 1 34.0' 133.6' 70.32. 148.50 180.41E 185.12E 84.00' B * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T ( P <0.05). 1 DATA R E P R E S E N T T H E T O T A L O B T A I N E D F R O M T H E R I G H T A N D L E F T WET G A S T R O C N E M I U S M U S C L E P E R C H I C K S A M P L E D . DATA R E P R E S E N T M G . / G . WET T I S S U E , TABLE 4 . - - L I V E R W E I G H T A N D DNA C O N T E N T F R O M C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E F O R TWO W E E K S L Y S 1NE T R E A T M E N T [%) L I V E R WT. ^  L I V E R DNA* S U P P . L E V E L G * G./100G. B O D Y WT.** MG,/G.' M G . / L I V E R MG./100G. B O D Y WT. 0.25 0.50 0.75 1 .00 0.36 0.61 0.86 1.11 1 .36 1.4 A 3.3 B C 4.1° 4.2 C 2.5 B 2.2 A 3.3 B 3.5 B 3.4 B 3.4 B 1 .9 A 2.3 B 2.6 B C 2.8° 2.9 C 2.7 A 7.6 B 10.7 C 11 .6 C 7.2 B 4.2 A 7.4 B 9.2° 9.3° 9.9 C * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P <0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P «=0.01). 1 DATA R E P R E S E N T M G . / G . WET T I S S U E . TABLE 5 . - - L I V E R RNA A N D P R O T E I N C O N T E N T F R O M C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E F O R TWO W E E K S RNA* P R O T E I N * L Y S I N E T R E A T M E N T {%) S U P P . L E V E L / 1 MG ./G . M G . / L I V E R M G . / 1 0 0 G . B O D Y WT. / 1 M G . / G . M G . / L 1 V E R M G . / 1 0 0 G . B O D Y WT.** 0.36 0.25 0.61 0.50 0.86 0.75 1.11 1.00 1 .36 7 5 A ' A 7.9* 9.5 B 7.7 A 7.3* 10.8* 26.3 8 39.0 C 32.4 C 18.4* 8 16.5* 25.5 B 33.6 26.2 B 24.9 B 146.2* 148.9* 166.1* 158.7* 156.4* 210.0* 496.1 8 677.9 C 659.8 C 393.4 B 321.2* 485.9 B 587.2 B 536.0 B 533.3 B * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F 1 C A N T L Y D 1 F F E R E N T (P < 0.05). **V/ALUES F O L L O W E D B Y A D I F F E RE N T S U P E R S C R 1 P T A R E S 1 G N 1 F 1 C A N T L Y D I F F E R E N T (P < 0.01). ^DATA R E P R E S E N T M G . / G . WET T I S S U E L Y S I N E S U P P L E M E N T A T I O N D E M O N S T R A T E D T H E L I M I T A T I O N O F P O O R Q U A L I T Y W H E A T P R O T E I N W H E N F E D A S A S O L E S O U R C E O F P R O T E I N TO C H I C K S . M U S C L E W E I G H T R E F L E C T E D P O O R G R O W T H O F C H I C K S F E D T H E 0.36$ L Y S I N E D I E T ( T A B L E 6 ) . THE R E S U L T A G R E E S W I T H T H E R E P O R T T H A T U N D E R N U T R I T I O N O F T H E C H I C K F R O M A N E A R L Y A G E G R O S S L Y R E T A R D E D T H E I N C R E A S E I N B O D Y W E I G H T A N D M U S C L E W E I G H T ( M O N T G O M E R Y _ET _ A L . 1 964). A N E X C E S S O F D I E T A R Y L Y S I N E D E P R E S S E D C H I C K G R O W T H ( T A B L E 6 ) . D E P R E S S E D C H I C K G R O W T H R E S U L T I N G F R O M L Y S I N E D E F I C I E N C Y (0.36$) A N D T O X I C I T Y (1.36$) WAS A T T R I B U T E D TO A D E C R E A S E I N F E E D I N T A K E ( T A B L E 6) A S P R E V I O U S L Y R E P O R T E D ( H A R P E R A N D C O - W O R K E R S , 1955, 1965; S M I T H , 1968; MARCH A N D W A L K E R , 1970; BRAGG _ET _ A L . 1971). F E E D I N T A K E B Y C H I C K S F E D T H E W H E A T - B A S A L D I E T WAS LOW T H R O U G H -O U T T H E S T U D Y ( F l G . 1) W H E R E A S C H I C K S F E D E X C E S S L Y S I N E C O N S U M E D MORE F E E D F O R ILIHE F I R S T S I X TO E I G H T D A Y S F O L L O W E D B Y A D E C L I N E I N F E E D I N T A K E C O M P A R E D TO C H I C K S R E C E I V I N G N O R M A L D I E T A R Y L Y S I N E (0.86$). T H E R A P I D D E C R E A S E I N F E E D I N T A K E W I T H C H I C K S F E D T H E D E F I C I E N T D I E T (0.36$) D E M O N S T R A T E S A N I M M E D I A T E I M P A C T ON T H E M E T A B O L I C S Y S T E M W H E R E A S T H E D E L A Y E D R E S P O N S E I N F E E D I N T A K E O F C H I C K S F E D E X C E S S L Y S I N E I N D I C A T E S A N I N H I B I T I N G M E C H A N I S M A U G M E N T E D P R O B A B L Y B Y A S E C O N D L I M I T I N G A M I N O A C I D . H O W E V E R , F E E D C O N S U M P T I O N WAS O B S E R V E D TO D E C R E A S E W I T H I N S I X TO E I G H T H O U R S O F F E E D I N G A L Y S I N E D E F I C I E N T D I E T TO C H I C K S B Y M A R C H A N D W A L K E R (1970). THE L Y S I N E R E Q U I R E M E N T F O R O P T I M U M C H I C K W E I G H T G A I N A N D G A I N / F E E D R A T I O WAS W I T H 0.86$ I N T H E 15$ P R O T E I N D I E T (0.5$ S U P P L E -M E N T A R Y L Y S I N E TO W H E A T - B A S A L ) . E X P R E S S I O N O F A M I N O A C I D R E Q U I R E M E N T S A S A U N I T O F W E I G H T P E R A N I M A L P E R D A Y I S M O R E P R E C I S E T H A N A S A TABLE 6.--BODY W E I G H T G A I N A N D F E E D I N T A K E O F C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E AGE 2 WEEKS 4 WEEKS  L Y S I N E T R E A T M E N T {%) WT. G A I N / C H I C K F E E D I N T A K E / C H I C K WT. G A I N / C H I C K F E E D I N T A K E / C H I C K S U P P . L E V E L G * G * G** G** 0.25 0.50 0.75 1 .00 0.36 0.61 0.86 1.11 1 .36 26 67 E 97 C 81 C 54 E 92 174 211' 191 156 E BC CD 56' 216 E 276 C 210 E 217 583 704 C 547 E B * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P "=0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P <=0.01). 61 AGE ( D A Y ) F I G . 1 F E E D I N T A K E P A T T E R N OP C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E F O R TWO W E E K S . P E R C E N T A G E O F T H E D I E T F R O M T H E P H Y S I O L O G I C A L V I E W P O I N T . H O W E V E R , C H A N G E S O B S E R V E D I N D A I L Y F E E D I N T A K E A S A R E S U L T O F G R O W T H ( A G E ) A N D T H E A C C E P T E D M E T H O D O F F E E D I N G C H I C K S I N L A R G E G R O U P S M A K E I T E S S E N T I A L TO E X P R E S S R E Q U I R E M E N T I N T E R M O F D I E T A R Y C O N T E N T . THE L Y S I N E R E Q U I R E M E N T F O R O P T I M U M G R O W T H A N D F E E D E F F I C I E N C Y WAS 129.3 M G / C H I C K / D A Y ( F I G . 2 ) . T H E S E R E S U L T S C O N F I R M R E P O R T S T H A T M A X I M A L G R O W T H I N T H E R A T I S O B T A I N E D W I T H A D I E T A R Y P R O T E I N L E V E L O F 16$ A N D A L Y S I N E L E V E L O F 0.84$ ( B R E S S A N I A N D M E R T Z , 1958) A N D I N T H E C H I C K W I T H A D I E T A R Y L E V E L O F 15$ O F P R O T E I N A N D 0.86$ O F L Y S I N E ( B R A G G _ET AJ_. 1971). G A S T R O C N E M I U S M U S C L E DNA, RNA A N D P R O T E I N C O N T E N T ( T A B L E 3) E X P R E S S E D A S M G . / G . WET T I S S U E D I D NOT C H A N G E S I G N I F I C A N T L Y W I T H D I E T A R Y L Y S I N E C O N T E N T . AN I N C R E A S I N G T R E N D I N M U S C L E RNA A N D P R O T E I N C O N T E N T ( M G . / G . WET T I S S U E ) , H O W E V E R , WAS E V I D E N T B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36 TO 0.61 A N D 0.86$. A D E C L I N I N G P A T T E R N I N T H E M U S C L E C O N S T I T U E N T S R E S U L T E D B E Y O N D 0.86$ D I E T A R Y L Y S I N E . DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N W H E N E X P R E S S E D P E R T O T A L M U S C L E W E I G H T F O L L O W E D E S S E N T I A L L Y T H E S A M E P A T T E R N A S T H E O R G A N W E I G H T , I N C R E A S I N G S I G N I F I C A N T L Y (P <0.05) W I T H M O D E R A T E A N D A D E Q U A T E L E V E L S OF D I E T A R Y L Y S I N E A N D D E C L I N I N G W I T H H I G H L E V E L O F L Y S I N E (1.36$). S I N C E C H I C K S U S E D I N T H E S T U D Y W E R E A L M O S T I D E N T I C A L I N I N I T I A L B O D Y W E I G H T S , T H E N U C L E I C A C I D S A N D P R O T E I N C O N T E N T S O F T H E G A S T R O C N E M I U S M U S C L E W E R E C O M P A R E D D I R E C T L Y B Y E X P R E S S I N G DNA, RNA A N D P R O T E I N A S T O T A L P E R C H I C K M U S C L E . A B S O L U T E M U S C L E DNA, RNA A N D P R O T E I N C O N T E N T A T D I F F E R E N T D I E T A R Y L Y S I N E L E V E L S R E F L E C T E D M U S C L E G R O W T H I N P R O P O R T I O N TO T H E I N C R E A S E I N B O D Y W E I G H T ( T A B L E S 3 A N D 6 ) . |T WAS R E C E N T L Y R E P O R T E D F I G . 2. A V E R A G E D A I L Y G A I N S A N D F E E D E F F I C I E N C I E S O F G R A D E D L E V E L S O F L Y S I N E F O R TWO W E E K S . C H I C K S F E D ( H O W A R T H , 1972) T H A T B O D Y W E I G H T , M U S C L E W E I G H T , DNA, RNA A N D P R O T E I N C O N T E N T I N C R E A S E D W I T H D I E T A R Y P R O T E I N I N T H E R A T . M U S C L E C E L L S I Z E , M E A S U R E D B Y P R O T E I N / D N A A N D RNA/DNA R A T I O S ( F l G . 3) R E F L E C T E D T H E I M P A C T O F D I E T A R Y L Y S I N E ON C H I C K G R O W T H ( T A B L E 6 ) . THE R A T I O S I M P R O V E D B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36 TO 0.61 A N D 0.86$, F O L L O W E D B Y A D E C L I N E W I T H F U R T H E R I N C R E M E N T O F L Y S I N E . B A N K S E T A L . (1964) O B T A I N E D S I M I L A R R E S U L T S W I T H R A T S B Y S U P P L E M E N T I N G W H E A T G L U T E N W I T H L Y S I N E A N D S U G G E S T E D T H A T F R E E L Y S I N E A D D E D TO T H E W H E A T G L U T E N WAS U T I L I Z E D TO I N C R E A S E M U S C L E G R O W T H . THE C U R V E L I N E A R I N C R E A S E I N M U S C L E P R O T E I N / D N A A N D RNA/DNA R A T I O S W I T H I N C R E A S I N G L E V E L O F D I E T A R Y L Y S I N E D E M O N S T R A T I N G A N I N C R E A S E I N M U S C L E C E L L S I Z E I N P R O P O R T I O N TO C H A N G E S I N B O D Y W E I G H T A L S O A G R E E S W I T H T H E R E P O R T S T H A T T H E R E I S A C L O S E A S S O C I A T I O N B E T W E E N M U S C L E F I B R E S I Z E A N D B O D Y S I Z E A T V A R I O U S A G E S I N C H I C K E N S ( M E H N E R , 1938), A N D T H A T T H E R E A R E H I G H L Y S I G N I F I C A N T P O S I T I V E C O R R E L A T I O N S B E T W E E N M E A N M U S C L E C E L L D H A M E T E R A N D B O D Y W E I G H T O F T H E R A B B I T , S W I N E , S H E E P A N D C A T T L E (MCMEEKAN, 1940-41; J O U B E R T , 1956), A N D B E T W E E N T H E N I T R O G E N C O N T E N T O F T H E H I N D L I M B A N D T H A T O F T H E W H O L E C A R C A S S I N T H E R A T ( L A C H A N C E A N D M I L L E R , 1973). T H E C U R V E L I N E A R I N C R E A S E I N M U S C L E P R O T E I N / D N A A N D RNA/DNA R A T I O S W I T H I N C R E A S I N G L E V E L O F D I E T A R Y L Y S I N E D E M O N S T R A T E S A H Y P E R T R O P H I C S T A T E O F M U S C L E C E L L S S I N C E I T I S B E L I E V E D T H A T RNA I S T H E B I O C H E M I C A L C O U N T E R P A R T O F H Y P E R T R O P H Y A N D DNA T H A T O F H Y P E R P L A S I A ( A L H O , 1961). L I V E R DNA C O N T E N T E X P R E S S E D A S M G . / G . WET T I S S U E OR MG./100G. B O D Y W E I G H T I N C R E A S E D P R O G R E S S I V E L Y W I T H D I E T A R Y L Y S I N E C O N T E N T ( T A B L E 4 ) . T H E V A L U E S O B T A I N E D W I T H 0.86, 1.11 A N D 1.36$ D I E T A R Y 65 O or 250 225 z Q < Z F I G . 3. G A S T R O C N E M I U S M U S C L E P R O T E I N / D N A A N D RNA/DNA R A T I O S I N C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E F O R TWO W E E K S . L Y S I N E , H O W E V E R , D I D N O T D I F F E R S I G N I F I C A N T L Y . DNA C O N C E N T R A T I O N W H E N E X P R E S S E D P E R T O T A L L I V E R W E I G H T R E F L E C T E D C H A N G E S I N O R G A N W E I G H T W I T H T H E A M O U N T O F L Y S I N E I N T H E D I E T A S I N T H E M U S C L E . C A M P B E L L A N D K O S T E R L I T Z (1950) H A V E D E M O N S T R A T E D T H A T DNA C O N T E N T I N R A T L I V E R I N C R E A S E D A S T H E B O D Y W E I G H T I N C R E A S E D . T H E I N C R E A S E D L I V E R DNA (MG./G. WET T I S S U E OR MG./1OOG. B O D Y W E I G H T ) W I T H D I E T A R Y L Y S I N E I N D I C A T E S , A N I N C R E A S E D C E L L N U M B E R ( G R O W T H ) I N T H E O R G A N . IN T H E I N T E R P R E T A T I O N O F D A T A F O R L I V E R DNA, I N T E R M S O F T H E C O M P O S I T I O N P E R G R A M O F WET T I S S U E , I T I S R E C O G N I Z E D T H A T T H E L I V E R H A S C E L L S W H I C H D E V E L O P F R O M T H E D I P L O I D S T A G E TO T H E T E T R A P L O I D OR O C T A P L O I D S T A T E B Y R E P L I C A T I O N O F T H E DNA W I T H O U T D I V I S I O N O F T H E N U C L E U S ( E P S T E I N , 1967). THE O C C U R R E N C E O F S U C H C E L L S I N .THE F O W L ' S L I V E R , H O W E V E R , H A S N O T B E E N R E P O R T E D . M O R E O V E R , M U R A M A T S U E T A L . (1971) O B S E R V E D T H A T R A T S F E D H I G H L E V E L S O F L Y S I N E HAD L I V E R S S L I G H T L Y H I G H E R I N DNA W H E N T H E P A R A M E T E R WAS E X P R E S S E D A S M G . / G . WET T I S S U E A N D L O W E R T O T A L DNA P E R L I V E R A B S O L U T E W E I G H T . L I V E R RNA A N D P R O T E I N C O N T E N T ( T A B L E 5) R E S P O N D E D T O D I E T A R Y L Y S I N E I N A S I M I L A R M A N N E R ; I N C R E A S I N G P R O G R E S S I V E L Y A S D I E T A R Y L Y S I N E I N C R E A S E D F R O M 0.36$ TO 0.61 A N D 0.86$ A N D D E C L I N I N G W I T H H I G H E R L E V E L S O F L Y S I N E . A S I G N I F I C A N T L Y (P -=0.05) H I G H E R C O N C E N T R A T I O N O F RNA (MG./G. WET T I S S U E A N D MG./lOOG. B O D Y W E I G H T ) WAS O B T A I N E D I N T H E L I V E R O F C H I C K S F E D F O R TWO W E E K S T H E 0.86$ L Y S I N E D I E T T H A N I N T H E L I V E R S O F C H I C K S R E C E I V I N G E I T H E R L O W E R OR H I G H E R L E V E L S O F D I E T A R Y L Y S I N E . T O T A L L I V E R RNA ( MG . / L I V E R), , H O W E V E R , I N C H I C K S F E D 0.86 A N D 1.11$ L Y S I N E D I D N O T D I F F E R S I G N I F I C A N T L Y B U T S I G N I F I C A N T L Y (P «= 0.05) H I G H E R C O N C E N T R A T I O N O F RNA ( M G . / G . WET T I S S U E A N D M G . / l O O G . B O D Y W E I G H T ) WAS O B T A I N E D I N T H E L I V E R O F C H I C K S F E D F O R TWO W E E K S T H E 0.86$ L Y S I N E D I E T T H A N I N T H E L I V E R S O F C H I C K S R E C E I V I N G E I T H E R L O W E R OR H I G H E R L E V E L S O F D I E T A R Y L Y S I N E . L I V E R P R O T E I N E X P R E S S E D A S M G . / G . WET T I S S U E I N C R E A S E D I N A C U R V E L I N E A R P A T T E R N T H A T WAS N O T S I G N I F I C A N T W I T H I N C R E A S I N G D I E T A R Y L Y S I N E . BY E X P R E S S I N G L I V E R P R O T E I N A S MG./LOOG. B O D Y W E I G H T , T H E P A R A M E T E R I N C R E A S E D S I G N I F I C A N T L Y (P <0.01) W H E N D I E T A R Y L Y S I N E WAS I N C R E A S E D F R O M 0.36 TO 0.61$. NO S I G N I F I C A N T I N C R E A S E I N T H E P A R A M E T E R O C C U R R E D B E Y O N D 0.61$ D I E T A R Y L Y S I N E . A B S O L U T E L I V E R P R O T E I N ( M G . / L I V E R ) I N C H I C K S F E D 0.86 A N D 1.11$ L Y S I N E WAS S I G N I F I C A N T L Y (P -=0.05) H I G H E R T H A N I N C H I C K S R E C E I V I N G 0.36, 0.61 A N D 1.36$ L Y S I N E D I E T S B U T D I D N O T D I F F E R F R O M E A C H O T H E R S I G N I F I C A N T L Y . T H E S E R E S U L T S ON L I V E R P R O T E I N (MG./100G. B O D Y W E I G H T A N D M G . / L I V E R ) A R E S I M I L A R TO D A T A ON L I V E R W E I G H T ( T A B L E 4) A N D I N D I C A T E T H A T L I V E R P R O T E I N WAS I N F L U E N C E D B Y L I V E R S I Z E W H I C H I N C R E A S E D P R O P O R T I O N A T E L Y W I T H I N C R E A S E S I N B O D Y W E I G H T O F C H I C K S F E D L Y S I N E S U P P L E M E N T E D W H E A T D I E T S . HOWEVER, L I V E R S I Z E ^ T A B L E 4) A N D L I V E R P R O T E I N ( T A B L E 5) D I D N O T A P P E A R TO C H A N G E R E L A T I V E TO B O D Y W E I G H T I N T H E L Y S I N E D E F I C I E N T (0.36$) G R O U P . R E S U L T S O F T H E P R E S E N T S T U D Y A G R E E W I T H P R E V I O U S R E P O R T S , V A N D E R M E E R S - P I R E T _ET _AL . (1966) O B S E R V E D T H A T R A T S D E P R I V E D O F L Y S I N E A N D T H R E O N I N E HAD L O W E R L I V E R P R O T E I N A N D RNA C O M P A R E D W I T H T H O S E I N C O N T R O L R A T S R E C E I V I N G 18$ C A S E I N A D E Q U A T E I N L Y S I N E A N D T H R E O N I N E . THE L I V E R P R O T E I N A N D RNA C O N C E N T R A T I O N I N R A T S F E D A H I G H L E V E L O F L Y S I N E I N A LOW P R O T E I N D I E T WERE A L S O O B S E R V E D ( M U R A M A T S U E T A L • 1971) N O T T O D I F F E R S I G N I F I C A N T L Y F R O M T H O S E O F T H E C O N T R O L G R O U P W H E N T H E P A R A M E T E R S W E R E E X P R E S S E D A S M G . / G . WET T I S S U E . H O W E V E R , T O T A L L I V E R P R O T E I N A N D RNA O F A H I G H L Y S I N E S U P P L E M E N T E D G R O U P W E R E L O W E R T H A N T H O S E O F T H E C O N T R O L G R O U P S . IN T H E E X P R E S S I O N O F L I V E R P R O T E I N A N D RNA, S O M E D I F F I C U L T I E S A R E O F T E N E N C O U N T E R E D . FOR E X A M P L E , T H E L I V E R W E I G H T C A N V A R Y W I T H I N W I D E L I M I T S B E C A U S E OF G A I N OR L O S S O F G L Y C O G E N A N D N E U T R A L F A T ( K O S T E R L I T Z A N D C A M P B E L L , 1945). WHEN M E A S U R E M E N T S A R E M A D E ON T I S S U E P R O T E I N S T H E C O M P O S I T I O N O F T H E T I S S U E S M A Y NOT B E T H E S A M E I N T H E G R O U P S O F A N I M A L S B E I N G C O M P A R E D , S I N C E A L O W - P R O T E I N D I E T C A U S E S A S E L E C T I V E L O S S O F O N E OR MORE T Y P E S O F P R O T E I N . AN E X A M P L E I S T H E R E D U C T I O N I N C O N C E N T R A T I O N O F P L A S M A A L B U M I N , B U T N O T O F P L A S M A G L O B U L I N S , P R O D U C E D B Y P R O T E I N D E F I C I E N C Y B O T H I N A N I M A L S A N D I N M A N ( W A T E R L O W _ET _ A L . 1960). TWO A P P R O A C H E S I N T H E L I T E R A T U R E A R E U S E D TO O V E R C O M E T H I S D I F F I C U L T Y A N D A R E A S F O L L O W S : (1) L I V E R P R O T E I N OR RNA I S E X P R E S S E D A S A U N I T OF B O D Y W E I G H T ; A N D (2) L I V E R C O N S T I T U E N T S A R E E X P R E S S E D P E R U N I T O F L I V E R DNA. BOTH P R O T E I N / D N A A N D RNA/DNA R A T I O S I N C H I C K L I V E R ( F I G . 4) D E C R E A S E D W I T H I N C R E A S I N G D I E T A R Y L Y S I N E A T 14 D A Y S O F A G E . R E S U L T S W I T H T H E L I V E R S U G G E S T TWO C O N D I T I O N S : (1) L I V E R G R O W T H I N C H I C K S A T 14 D A Y S O F A G E I S M A I N L Y B Y A N I N C R E A S E I N C E L L N U M B E R ( H Y P E R P L A S I A ) . L I T T L E C H A N G E O C C U R S I N C E L L S I Z E S I N C E P R O T E I N / D N A A N D RNA/DNA R A T I O S A R E M E A S U R E M E N T S O F C E L L S I Z E (M-UNRO A N D F L E C K , 1966). T H E . V I E W I S S U P P O R T E D B Y A S I G N I F I C A N T 69 DIETARY LYSINE {%) FIG. 4. LIVER PROTEIN/DNA, RNA/DNA AND PROTEIN/RNA RATIOS FROM CHICKS FED GRADED LEVELS OF DIETARY LYSINE FOR TWO WEEKS . (P < 0.05) I N C R E A S E I N L I V E R DNA (MG./G. WET T I S S U E ) C O N C E N T R A T I O N W I T H I N C R E A S I N G D I E T A R Y L Y S I N E ( T A B L E 4). (2) C E L L U L A R L I V E R P R O T E I N S I N L Y S I N E D E F I C I E N T C H I C K S P R O B A B L Y R E P R E S E N T E D R E S I D U A L P R O T E I N S S U P P L I E D B Y T H E E G G D U R I N G E M B R Y O N I C D E V E L O P M E N T A N D E A R L Y P O S T - H A T C H . M O R E O V E R , I T WAS O B S E R V E D I N T R I A L 4 T H A T L I V E R P R O T E I N S W E R E D E G R A D E D A N D S Y N T H E S I Z E D A T S L O W E R R A T E S I N C H I C K S F E D T H E W H E A T - B A S A L D I E T C O N T A I N I N G 0.36$ L Y S I N E T H A N I N C H I C K S R E C E I V I N G A D E Q U A T E L Y S I N E (0.86$). H A R P E R (1965, 1968) H A S R E P O R T E D T H A T G R O W T H R A T E A N D P R O T E I N S Y N T H E S I S A R E LOW I N A N I M A L S F E D A LOW P R O T E I N D I E T . I T I S U N L I K E L Y T H A T M U S C L E P R O T E I N WAS C A T A B O L I Z E D TO S U P P L Y A M I N O A C I D S I N T O T H E M E T A B O L I C P O O L W H I C H C O U L D I N C R E A S E L I V E R P R O T E I N S S I N C E M U S C L E P R O T E I N S P E C I F I C A C T I V I T Y I N L Y S I N E D E F I C I E N T C H I C K S D I D N O T D I F F E R M A R K E D L Y A T 8 A N D 15 D A Y S OF A G E ( T R I A L 4, F I G . 22). THE L I V E R P R O T E I N / R N A R A T I O WAS C O N S T A N T (FIG. 4) I R R E S P E C T I V E L Y O F D I E T A R Y L Y S I N E C O N T E N T . R E S U L T S O F T H I S T R I A L C O N F I R M T H E O B S E R V A T I O N O F K O S T E R L I T Z (1947) A N D A L L I S O N _ET _AL . (1964) W I T H R A T S T H A T L I V E R P R O T E I N A N D RNA C O N T E N T W E R E C O R R E L A T E D . THE G A S T R O C N E M I U S M U S C L E A N D L I V E R O F C H I C K S R E C E I V I N G 0.36, 0.61, 0.86 A N D 1.iH$ D I E T A R Y L Y S I N E F O R F O U R W E E K S W.E'R:E A N A L Y Z E D F O R DNA, RNA A N D P R O T E I N . M U S C L E W E I G H T A N D T H E B I O C H E M I C A L C O M P O S I T I O N A R E P R E S E N T E D I N T A B L E 7. L l V E R W E I G H T A N D T H E DNA C O N T E N T , RNA A N D P R O T E I N C O N C E N T R A T I O N A R E R E S P E C T I V E L Y I L L U S T R A T E D I N T A B L E S 8 A N D 9. M U S C L E W E I G H T I N C R E A S E D P R O G R E S S I V E L Y (P -=0.05) B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36$ TO 0.61 A N D 0.86$ A N D D E C L I N E D W I T H 1.11$ L Y S I N E . M U S C L E DNA A N D RNA E X P R E S S E D A S M G . / G . WET T I S S U E W E R E NOT TABLE 7 . — G A S T R O C N E M I U S M U S C L E W E I G H T , DNA, RNA A N D P R O T E I N C O N T E N T F R O M C H I C K S F E D G R A D E D L E V E L S OF D I E T A R Y L Y S I N E F O R F O U R W E E K S M U S C L E L Y S I N E T R E A T M E N T (%) WT. DNA RNA* P R O T E I N ! • — 1 * - 7 —2* 1** 7 — 2 1 7 — 2 * S U P P . L E V E L MG. M G . / G . MG. M G . / G . MG. M G . / G . 0.36 1018.2 A 0.67 A 0.69 A 1.17 A 1.20 A 147.3 A 149.92 A 0.25 0.61 3922.7 B 0.63 A 2.47 B 1.50 A 5.88 B 200.7 B 787.32 B 0.50 0.86 4547.5° 0.53 A 2.40 B 1.12 A 5.12 B 178.3° 812.68 B 0.75 1.11 3498.2 B 0.71 A 2.20 B 1.28 A 4.02° 192.2 B C 673.92 B * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P <0 . 0 5 ) . * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P < 0.01). 1 DATA R E P R E S E N T T H E T O T A L O B T A I N E D F R O M T H E R I G H T A N D L E F T WET G A S T R O C N E M I U S M U S C L E P E R C H I C K S A M P L E D . 2 R "DATA R E P R E S E N T M G . / G . WET T I S S U E TABLE 8 . - - L I V E R W E I G H T A N D DNA C O N T E N T F R O M C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E F O R F O U R W E E K S LI I V E R W T . DNA L Y S 1 N E S U P P . T R E A T M E N T [%) L E V E L G.** G . / 1 0 0 G B O D Y WT 1 * .* M G . / G . M G . / L I V E R * * M G . / 1 0 0 G . B O D Y WT. 0.25 0.50 0.75 0.36 0.61 0.86 1.11 2 . 9 A 9.8 B 1 2 . 1 B 9.7 B 2 . 7 A 3 .1 A 3.3 A 3 . 0 A 2 . 5 A 2 . 2 A 2 . 2 A 2 . 3 A A 7 . 2 21 .5 B 26.7 B 21 .9 B A * 6.7 6.8 A 7.3 A 6.9 A ^ V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P « c 0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P " ^ O . O l ) . DATA R E P R E S E N T M G . / G . WET T I S S U E . TABLE 9.--LIVER RNA AND PROTEIN CONTENT FROM CHICKS FED GRADED LEVELS OF DIETARY LYSINE FOR FOUR WEEKS RNA PROTEIN LYSINE TREATMENT {%) SUPP. LEVEL / 1* MG ./L1VER** MG . / 1 0 0 G . BODY WT.* / 1 * MG ,/G . MG ,/lIVER** MG . / 1 0 0 G . BODY WT.* 0.36 0.25 0.61 0.50 0.86 0.75 1.11 6.3 A A 7.0 7.0 A 6.2 A 18.5 A 69.4 B 85.2 B 60.4 B 17.0 A 22.2 A 23.2 A 1 8.8 A 137.4 A 139.7 A 144.7 A 140.4 A 398.5 A 1377.6 8 1749.6 B 1352.3 8 368.9 A 440.4 A 475.4 423.3 A *VALUES FOLLOWED BY A DIFFERENT SUPERSCRIPT ARE S 1 GNI F1CANTLY D I FFERENT (P < 0.05). **VALUES FOLLOWED BY A DIFFERENT SUPERSCRIPT ARE S 1 GN1F1CANTLY DIFFERENT (P < 0 . 0 1 ) . 1 DATA REPRESENT MG./G . WET TISSUE 74 A F F E C T E D S I G N I F I C A N T L Y B Y D I E T A R Y L Y S I N E C O N T E N T . M U S C L E P R O T E I N C O N C E N T R A T I O N ( M G . / G . ) , H O W E V E R , I N C R E A S E D (P <0.05) W I T H D I E T A R Y L Y S I N E . DNA A N D P R O T E I N C O N T E N T O F T H E M U S C L E E X P R E S S E D A S M G . / M U S C L E S A M P L E I N C R E A S E D (P «= 0.01 ) WHEN D I E T A R Y L Y S I N E WAS I N C R E A S E D F R O M 0.36 TO 0.61$. A F U R T H E R A D D I T I O N O F L Y S I N E D I D N O T R E S U L T I N A S I G N I F I C A N T C H A N G E I N M U S C L E DNA A N D P R O T E I N C O N T E N T . T O T A L RNA C O N T E N T O F T H E G A S T R O C N E M I U S M U S C L E I N C R E A S E D (p < 0.05) W I T H M O D E R A T E A N D A D E Q U A T E D I E T A R Y L Y S I N E (0.61 A N D 0.86$) A N D D E C L I N E D W I T H H I G H E R L Y S I N E (1.11$). C H A N G E S I N G A S T R O C N E M I U S M U S C L E W E I G H T , DNA, RNA A N D P R O T E I N W I T H D I E T A R Y L Y S I N E C O N T E N T I N 28 D A Y - O L D C H I C K S A G R E E W I T H A N D C O N F I R M T H E D A T A C O L L E C T E D F R O M 14 D A Y - O L D C H I C K S W H I C H R E F L E C T E D B O D Y G R O W T H P A T T E R N O F T H E C H I C K I N R E S P O N S E T O A V A I L A B L E D I E T A R Y L Y S I N E ( T A B L E 6). PROTEIN/DNA R A T I O IN T H E G A S T R O C N E M I U S M U S C L E F O L L O W E D E S S E N T I A L L Y T H E P A T T E R N O B S E R V E D A T 14 D A Y S O F A G E ( F l G . 5). IT I N C R E A S E D W I T H 0.36 TO 0.86$ D I E T A R Y L Y S I N E A N D D E C L I N E D A T 1.11$ L Y S I N E L E V E L . H O W E V E R , T H E R A T I O WAS H I G H E R A T 28 D A Y S T H A N A T 14 D A Y S O F A G E I R R E S P E C T I V E L Y O F D I E T A R Y L Y S I N E T R E A T M E N T I N D I C A T I N G I N C R E A S E D M U S C L E P R O T E I N W I T H A G E . DlCKERSON (i960) O B S E R V E D T H A T B O T H T O T A L P R O T E I N N I T R O G E N A N D N O N - P R O T E I N N I T R O G E N I N C R E A S E D I N T H E C H I C K M U S C L E W I T H A G E . M U S C L E RNA/DNA R A T I O , I N C R E A S E D W I T H 0.61$ L Y S I N E A N D D E C L I N E D W I T H H I G H E R L E V E L O F L Y S I N E . THE R E S U L T S U G G E S T S A R A P I D D E G R A D A T I O N O F RNA B E C A U S E O F A C T I V E M U S C L E P R O T E I N S Y N T H E S I S A T H I G H E R L Y S I N E L E V E L . THE V I E W I S S U P P O R T E D B Y A N I N C R E A S E I N T H E P R O T E I N / R N A R A T I O A T D I E T A R Y L Y S I N E L E V E L H I G H E R T H A N 0.61$ ( F I G . 5). THE LOW FIG. 5. GASTROCNEMIUS MUSCLE PROTEIN/DNA, RNA/DNA AND PROTEIN/RNA RATIOS FROM CHICKS FED GRADED LEVELS OF DIETARY LYSINE FOR FOUR WEEKS. PROTEIN/RNA I N T H E M U S C L E O F C H I C K S F E D T H E W H E A T B A S A L ( C O N T A I N I N G 0.36$ L Y S I N E ) A S A S O L E S O U R C E O F P R O T E I N , P R O B A B L Y R E F L E C T E D A C O M P E N S A T I N G M E C H A N ' I S M W H E R E B Y T H E M U S C L E P R O T E I N WAS C A T A B O L I Z E D TO S U P P L Y A M I N O A C I D S F O R E S S E N T I A L P H Y S I O L O G I C A L P R O C E S S E S W H I C H C O U L D C O R R E C T T H E A M I N O A C I D I M B A L A N C E S U P P L I E D B Y W H E A T . L I V E R W E I G H T ( T A B L E 8) O F F O U R W E E K - O L D C H I C K S I N C R E A S E D (p < 0.01 B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36 TO 0.61$. No F U R T H E R I N C R E A S E O C C U R R E D I N L I V E R W E I G H T B E Y O N D 0.61$ D I E T A R Y L Y S I N E . H O W E V E R , L I V E R W E I G H T E X P R E S S E D A S G./100G. B O D Y W E I G H T WAS U N A F F E C T E D S I G N I F I C A N T L Y B Y D I E T A R Y L Y S I N E . L I V E R DNA ( T A B L E 8), RNA A N D P R O T E I N ( T A B L E 9) E X P R E S S E D A S M G . / G . WET T I S S U E WERE U N A F F E C T E D S I G N I F I C A N T L Y W I T H T H E C O N T E N T O F D I E T A R Y L Y S I N E . E X P R E S S I N G T H E P A R A M E T E R S A S M G . / L I V E R , H O W E V E R , R E S U L T E D I N A S I G N I F I C A N T (P ??0.01) I N C R E A S E W I T H D I E T A R Y L Y S I N E I N A M A N N E R S I M I L A R TO T H E P A T T E R N O F O R G A N W E I G H T . L I V E R G R O W T H O F C H I C K S A T F O U R W E E K S O F A G E U N D E R T H E P R E S E N T E X P E R I M E N T A L C O N D I T I O N S ( T A B L E 9) WAS B Y A N I N C R E A S E I N C E L L S I Z E . THE V I E W I S S U P P O R T E D B Y T H E C U R V E L I N E A R N A T U R E O F P R O T E I N / D N A A N D RNA/DNA R A T I O S W I T H D I E T A R Y L Y S I N E ( F I G . 6). PROTEIN/RNA R A T I O I N T H E L I V E R R E M A I N E D C O N S T A N T W I T H D I E T A R Y L Y S I N E A S WAS P R E V I O U S L Y O B S E R V E D A T TWO W E E K S O F S T U D Y . THE I N C R E A S E I N L I V E R DNA ( M G . / G . T I S S U E ) W I T H D I E T A R Y L Y S I N E I N T H E C H I C K A T TWO W E E K S O F A G E A N D N O T A T F O U R W E E K S S U G G E S T S A N A C C E L E R A T E D R A T E O F L I V E R C E L L M U L T I P L I C A T I O N OR G R O W T H E A R L Y I N T H E C H I C K L I F E . L I V E R G R O W T H R A T E WAS P R O B A B L Y D E C L I N I N G B Y T H E T I M E T H E C H I C K R E A C H E D F O U R W E E K S O F A G E . M E N D E S A N D WATERLOW (1958) S T U D I E D PROTEIN/RNA RNA/DNA PROTEIN/DNA o CT> s: 0 1— m X — m — < 7; 0 Fl CO 7\ 33 • co "0 "n 33 n O 0 0 _ Z > —— 0 O FI 0 O r~ Fl m ID < m 33 r -< co O |— 0 •< -q co > 0 Z z — O Fl Fl H "D ^ ^ > 33 33 O -< H Fl r" — -c Z CO ^ — z Fl S» "H 33 O > 33 H -n O 0 CO c 33 "H 33 O 2 -J -J T H E E F F E C T O F A L O W - P R O T E I N D I E T A N D O F R E F E E D 1 N G ON T H E L I V E R A N D M U S C L E C O M P O S I T I O N I N T H E W E A N L I N G R A T A N D O B S E R V E D T H A T R A T E S O F DNA A N D P R O T E I N F O R M A T I O N I N T H E L I V E R F O L L O W E D T H E P A T T E R N O F G R O W T H , F A S T A T F I R S T A N D S L O W E R L A T E R . |N T H E M U S C L E T H E O P P O S I T E H A P P E N E D S O T H A T THE-RE WAS A L A G I N T H E F O R M A T I O N O F NEW M U S C L E . E N E S C O A N D L E B L O N D (1962) D I V I D E D T I S S U E G R O W T H I N T H E R A T I N T O T H R E E P H A S E S . F I R S T , D U R I N G T H E I N I T I A L 17 D A Y S O F P O S T N A T A L L I F E A L L T H E O R G A N S A N D T I S S U E S O F T H E R A T G R E W B Y A N I N C R E A S E I N C E L L N U M B E R A N D W I T H O N L Y T R I V I A L C H A N G E S I N C E L L S I Z E . S E C O N D F R O M 17 TO A B O U T 34 OR 48 D A Y S , T H E I N T E N S I T Y O F C E L L M U L T I P L I C A T I O N S L O W E D DOWN A N D S O M E I N C R E A S E I N W E I G H T D U E TO I N C R E A S E I N C E L L S I Z E B E G A N TO A P P E A R . T H E T H I R D P E R I O D O F G R O W T H F R O M 48 TO 160 D A Y S WAS M A R K E D F U R T H E R B Y E X T E N S I V E G R O W T H I N C E L L S I Z E . T H E D I F F E R E N C E I N T H E R E S P O N S E O F M U S C L E A N D L I V E R P R O T E I N S TO D I E T A R Y L Y S I N E A T TWO W E E K S O F A G E WAS A C C O U N T E D F O R B Y T H E F A C T T H A T M U S C L E P R O T E I N S R E P R E S E N T S T O R E D P R O T E I N W H I L E L I V E R P R O T E I N S A R E M I X T U R E O F B O T H L A B I L E A N D S T O R E D P R O T E I N S . S U M M A R Y A N D C O N C L U S I O N THE I M P A C T O F D I E T A R Y L Y S I N E D E F I C I E N C Y A N D E X C E S S E S ON C H I C K B O D Y A N D T I S S U E G R O W T H WAS M E A S U R E D . W H E A T (16$ P R O T E I N ) A S A S O L E S O U R C E O F P R O T E I N A N D W H E A T S U P P L E M E N T E D W I T H G R A D E D L Y S I N E L E V E L S (0.25, 0.5, 0.75, 1.0$) W E R E F E D TO B R O I L E R C H I C K S S T A R T I N G A T O N E D A Y O F A G E D U R I N G A F O U R W E E K P E R I O D . G A S T R O C N E M I U S M U S C L E A N D L I V E R DNA, RNA A N D P R O T E I N C O N T E N T W E R E M E A S U R E D A T TWO A N D F O U R W E E K S O F A G E . O P T I M U M C H I C K G R O W T H WAS O B T A I N E D W I T H 0.85$ L Y S I N E (0.5$ L - L Y S I N E S U P P L E M E N T E D TO W H E A T P R O T E I N ) . T O T A L O R G A N DNA, RNA A N D P R O T E I N C O N T E N T I N C R E A S E D W I T H M O D E R A T E (0.61$) A N D A D E Q U A T E (0.86$) D I E T A R Y L Y S I N E A N D D E C L I N E D W I T H E X C E S S (1.11, 1.36$) L Y S I N E . G A S T R O C N E M I U S M U S C L E G R E W B Y A N I N C R E A S E I N C E L L S I Z E B O T H A T TWO A N D F O U R W E E K S O F A G E A S I N D I C A T E D B Y L A C K O F C H A N G E I N M U S C L E DNA ( M G . / G . ) A N D T H E I N C R E A S E I N P R O T E I N / D N A A N D RNA/DNA R A T I O S A T B O T H P E R I O D S O F M E A S U R E M E N T S W I T H D I E T A R Y L Y S I N E . I N C R E A S I N G P R O T E I N / R N A R A T I O I N T H E M U S C L E A T F O U R W E E K S W I T H L Y S I N E L E V E L S U G G E S T S P R O T E I N B I O S Y N T H E S I S A N D A C C U M U L A T I O N . THE L I V E R A P P E A R E D TO GROW B Y A N I N C R E A S E I N C E L L N U M B E R ( H Y P E R P L A S I A ) D U R I N G T H E F I R S T TWO W E E K S O F A G E A N D B Y I N C R E A S E S B O T H I N C E L L N U M B E R A N D C E L L S I Z E ( H Y P E R T R O P H Y ) F R O M TWO TO F O U R W E E K S O F A G E . T H I S V I E W I S D E M O N S T R A T E D B Y I N C R E A S E I N L I V E R DNA ( M G . / G . ) W I T H I N C R E A S I N G L Y S I N E A T TWO W E E K S A N D T H E R E L A T I V E S T A B I L I T Y A T F O U R W E E K S . T H E D E C L I N I N G P R O T E I N / D N A A N D RNA/DNA R A T I O S I N T H E L I V E R A T TWO W E E K S A N D R E L A T I V E I N C R E A S E S A T F O U R W E E K S S U P P O R T T H E S A M E V I E W . L I V E R P R O T E I N / R N A R A T I O A T B O T H TWO A N D F O U R W E E K S O F A G E WAS C O N S T A N T A N D I N D E P E N D E N T O F L Y S I N E I N T A K E I N D I C A T I N G T H A T RNA I S C L O S E L Y C O R R E L A T E D W I T H P R O T E I N B I O S Y N T H E S I S . TRIAL 2. E X C E S S L Y S I N E WAS D E M O N S T R A T E D TO D E P R E S S T O T A L M U S C L E A N D L I V E R DNA, RNA A N D P R O T E I N I N T R I A L 1. T H I S T R I A L WAS TO C O N F I R M T H E O B S E R V A T I O N O F T R I A L 1 A N D TO D E T E R M I N E T H E I N F L U E N C E O F A R G I N I N E ON T H E T I S S U E C O N S T I T U E N T S I N A L L E V I A T I N G T H E A D V E R S E E F F E C T O F E X C E S S L Y S I N E ON C H I C K G R O W T H . T A B L E S 10 A N D 11 I L L U S T R A T E T H E R E S P O N S E O F C H I C K M U S C L E A N D L I V E R R E S P E C T I V E L Y TO E X C E S S D I E T A R Y L Y S I N E A N D W I T H A R G I N I N E S U P P L E M E N T A T I O N . M U S C L E A N D L I V E R W E I G H T S A N D T H E DNA, RNA A N D P R O T E I N C O N S T I T U E N T S D E C R E A S E D (P «=0.05) B Y S U P P L E M E N T I N G T H E W H E A T D I E T (0.36$ L Y S I N E ) W I T H E X C E S S L Y S I N E (1.0 A N D 1.5$) D U R I N G T H E 14 D A Y P E R I O D ( O N E D A Y O F A G E TO 14 D A Y S O F A G E ) . R E S U L T S C O N F I R M D A T A O F T R I A L 1 A N D A G R E E W I T H T H E R E P O R T ( M U R A M A T S U E T _ A L . 1971) T H A T F E E D I N G T H E R A T A 10$ C A S E I N D I E T C O N T A I N I N G E X C E S S L Y S I N E (5$) F O R T H R E E W E E K S D E P R E S S E D L I V E R W E I G H T , T O T A L L I V E R DNA, RNA A N D P R O T E I N . M U S C L E A N D L I V E R P R O T E I N A N D RNA W E R E E X P R E S S E D P E R U N I T O F T I S S U E DNA. E X C E S S D I E T A R Y L Y S I N E D E P R E S S E D M U S C L E P R O T E I N / D N A A N D RNA/DNA R A T I O S M A R K E D L Y ( F I G . 7 ) . DATA SHOW T H E S A M E A D V E R S E E F F E C T O F E X C E S S D I E T A R Y L Y S I N E ON G R O W T H I N C E L L S I Z E I N T H E M U S C L E A S WAS O B S E R V E D I N T R I A L 1. T H E E F F E C T O F E X C E S S D I E T A R Y L Y S I N E WAS M O R E S E V E R E ON M U S C L E P R O T E I N / D N A T H A N ON M U S C L E RNA/DNA R A T I O ( F l G . 7 ) . S I N C E M U S C L E P R O T E I N S R E P R E S E N T R E S E R V E D P R O T E I N , P R O T E I N / D N A R A T I O P R O B A B L Y I N D I C A T E S P R E V I O U S C H A N G E S I N T H E O R G A N W H E R E A S RNA/DNA R A T I O R E F L E C T S C H A N G E S T H A T W E R E O C C U R I N G A T T H E T I M E O F M U S C L E R E M O V A L F R O M T H E C H I C K . L I V E R P R O T E I N / D N A A N D RNA/DNA R A T I O S W E R E M A R K E D L Y D E P R E S S E D B Y TABLE 10. — G A S T R O C N E M I U S M U S C L E DNA, RNA A N D P R O T E I N C O N T E N T O F C H I C K S F E D N O R M A L A N D E X C E S S L Y S I N E F O R TWO W E E K S A N D A R G I N I N E S U P P L E M E N T E D TO T H E E X C E S S L Y S I N E D I E T F R O M TWO TO F O U R W E E K S 0-2 WK.* 2-4 WK.* . D I E T A R Y 1 M U S C L E WT. DNA ^ RNA P R O T E I N M U S C L E WT. DNA ^ RNA P R O T E I N T R E A T M E N T MG. MG. + 0. 5$ L- 2 L Y S 1 N E £ 1271 .8A .3B 0.68A 2.30A 1 .06B 180.4A 4547 .5A 2.40A 5. 12 812. 7A + 1. 0$ L- L Y S 1 N E 632 0.34B 84.0B ---+ + 1. 0. Ofo OK L-ifo L Y S I N E 2 L-ARG1N1NE .3B 3630 .2B 2.24A 5. A 08 722. 9A + 1. 5$ L- L YS 1 NEL- 562 0.32B 0.83B 45.1C - — + + 1. 0. 5f0 29^  L-1% L Y S 1 N E o L-ARG1N1NE 2726 .3B 1 .95A 3. A 88 437. 7A * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T ( P < 0.05). WHEAT B A S A L C O N T A I N E D 0.36$ L Y S I N E . WHEAT B A S A L P L U S L Y S I N E WAS F E D F R O M D A Y O F A G E TO TWO W E E K S O F A G E ; A R G I N I N E WAS T H E N S U P P L E M E N T E D TO T H E E X C E S S L Y S I N E D I E T F R O M TWO TO F O U R W E E K S O F A G E . DATA R E P R E S E N T T H E T O T A L O B T A I N E D F R O M T H E R I G H T A N D L E F T WET G A S T R O C N E M I U S M U S C L E P E R C H I C K S A M P L E D . TABLE 11.--LIVER DNA, RNA A N D P R O T E I N C O N T E N T O F C H I C K S F E D N O R M A L A N D E X C E S S L Y S I N E F O R TWO W E E K S A N D A R G I N I N E S U P P L E M E N T E D TO T H E E X C E S S L Y S I N E D I E T F R O M TWO TO F O U R W E E K S 0-2 WK.* 2-4 WK.*  D I E T A R Y L I V E R WT. DNA RNA P R O T E I N L I V E R WT. DNA RNA P R O T E I N T R E A T M E N T G. M G . / L I V E R G. M G . / L I V E R + + 0.5$ 1 .0$ p L - L Y S 1 N E p L - L Y S 1 N E ^ 4 2 .1A .5B 10.7A 7.2B 39.0A 18.48 677.9A 393.4B 12. 1A 26.7A 85.2A 1749. 6A + + 1 .0$ 0 . 0 1 : I  2 3$ L - A R G I N I N E ~5~~0B 17.2B 368.1B 9. oB 20.8AB A 71 .3 1355. 6A + 1 .5$ L- L YS 1 N E 2 ~2B — -+ + 1 .5$ 0.29: L - L Y S I N E 3$ L - A R G I N I N E -• -- 7. 9B 15.7B 63.5A 1317. 4A * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P <=0.05). WHEAT B A S A L C O N T A I N E D 0.36$ L Y S I N E . 2 WHEAT B A S A L P L U S L Y S I N E WAS F E D F R O M D A Y O F A G E TO TWO W E E K S OF A G E , A R G I N I N E WAS T H E N S U P P L E M E N T E D TO T H E E X C E S S L Y S I N E D I E T F R O M TWO TO F O U R W E E K S O F A G E . 250L IG. 7. GASTROCNEMIUS MUSCLE PROTEIN/DNA AND RNA/DNA RATIOS AS AFFECTED BY EXCESS DIETARY LYSINE IN THE CHICK. EXCESS DIETARY LYSINE ( F l G . 8) AS OBSERVED I N TRIAL 1. S E V E R E LYSINE TOXICITY OCCURRED WITH 1.86$ DIETARY LYSINE (1.5$ SUPPLEMENT) RETARDING BODY A N D LIVER GROWTH I N T H E CHICK. CHICKS RECEIVING T H E DIET ADEQUATE (0.86$) I N LYSINE (0.5$ SUPPLEMENT) GAINED 97G. A N D HAD A LIVER WEIGHT O F 4.1 G . I N 14 DAYS WHEREAS CHICKS F E D T H E EXCESS LYSINE (1.86$) DIET (1.5$ SUPPLEMENT) GAINED ONLY 40G. I N BODY WEIGHT AND HAD A LIVER WEIGHT O F 2.2G . THE DEPRESSED LIVER GROWTH (CELL MULTIPLICATION) WITH SEVERE LYSINE (1.86$ OF THE DIET) TOXICITY RESULTED I N HIGHER LIVER PROTEIN/DNA AND RNA/DNA RATIOS THAN NORMAL (FIG. 8 ) . RESULTS WERE SIMILAR TO HIGH PROTEIN/DNA AND RNA/DNA RATIOS OBSERVED IN THE LIVER OF CHICKS FED DEFICIENT LYSINE DIET IN TRIAL 1 (FIG. 4 ) . RESULTS INDICATED THAT CHICK GROWTH RETARDED BY FEED RESTRICTION DUE TO EITHER LOW OR EXCESS LYSINE IN THE DIET WAS ACCOMPANIED BY HIGH LIVER CELLULAR PROTEINS. HIGH PROTEIN/DNA AND RNA/DNA RATIOS IN LYSINE DEFICIENT (0.36$) CHICKS ARE BELIEVED AS IN TRIAL 1 TO BE DUE TO (1) LACK OF INCREASE IN CELL NUMBER (GROWTH) AND (2) RESIDUAL PROTEINS SUPPLIED BY THE EGG DURING EMBRYONIC DEVELOPMENT AND EARLY POST HATCH. THE TURNOVER OF LIVER PROTEIN IN CHICKS FED ADEQUATE (0.86$) DIETARY LYSINE APPEARS TO OCCUR AT A GREATER RATE THAN IN CHICKS WITH A SLOW GROWTH RATE (TRIAL 4, FLG. 21). Low PROTEIN/DNA AND RNA/DNA RATIOS IN THE MUSCLE OF CHICKS CONSUMING EXCESS (1.86$) DIETARY LYSINE INDICATE THAT MUSCLE PROTEIN SYNTHESIS (MUSCLE GROWTH IN CELL SIZE) IS IMPAIRED BY FEEDING EXCESS LYSINE TO CHICKS. THIS VIEW IS CONSISTENT WITH THE DEPRESSION IN FEED INTAKE OF CHICKS RECEIVING EXCESS (1.86$) DIETARY LYSINE COMPARED TO THE GROUP FED ADEQUATE (0.86$) DIETARY LYSINE. MOREOVER, UNDER THE CONDITION OF IG. 8. LIVER PROTEIN/DNA AND RNA/DNA RATIOS AS AFFECTED BY EXCESS DIETARY LYSINE IN THE CHICK. LOW F E E D I N T A K E , T H E C H I C K P R O B A B L Y D E G R A D E D M U S C L E P R O T E I N TO P R O V I D E A M I N O A C I D S F O R T H E S Y N T H E S I S O F P R O T E I N S V I T A L T O L I F E . G A S T R O C N E M I U S M U S C L E A N D L I V E R W E I G H T S D I D N O T SHOW A S I G N I F I C A N T R E S P O N S E TO T H E A D D I T I O N O F A R G I N I N E T O D I E T S C O N T A I N I N G E X C E S S L Y S I N E F R O M TWO TO F O U R W E E K S O F A G E . A B S O L U T E L I V E R DNA S H O W E D A N I N C R E A S E D T R E N D . M U S C L E DNA, RNA A N D P R O T E I N C O N T E N T I N C R E A S E D W I T H A R G I N I N E S U P P L E M E N T A T I O N T O V A L U E S N OT S I G N I F I C A N T L Y D I F F E R E N T F R O M T H O S E O F C H I C K S C O N S U M I N G A D E Q U A T E L Y S I N E ( T A B L E 10). THE I N C R E A S E I N P R O T E I N A N D RNA C O N C E N T R A T I O N I N T H E M U S C L E U P O N A D D I N G A R G I N I N E TO D I E T S C O N T A I N I N G E X C E S S (1.36, 1.86$) L Y S I N E A N D I N T H E L I V E R U P O N A D D I N G A R G I N I N E TO T H E 1.36$ L Y S I N E D I E T WAS A L S O R E F L E C T E D A T T H E C E L L U L A R L E V E L . T H I S V I E W I S I N D I C A T E D B Y A N I N C R E A S E I N P R O T E I N / D N A A N D RNA/DNA R A T I O S I N B O T H O R G A N S D U R I N G A R G I N I N E S U P P L E M E N T A T I O N R E L A T I V E TO R A T I O S O B T A I N E D W I T H O U T A R G I N I N E A D D I T I O N T O T H E A D E Q U A T E L Y S I N E D I E T ( F l G . 9 A N D 10). THE A B S O L U T E A M O U N T S O F DNA, RNA A N D P R O T E I N A C C U M U L A T E D D U R I N G A R G I N I N E S U P P L E M E N T A T I O N ( T WO TO F O U R W E E K S ) TO D I E T S C O N T A I N I N G E X C E S S L Y S I N E (1.36, 1.86$) W E R E E S T I M A T E D B Y D I F F E R E N C E S B E T W E E N V A L U E S A T 14 A N D 28 D A Y S O F A G E . A C C U M U L A T E D M U S C L E A N D L I V E R C O N S T I T U E N T S A N D B O D Y W E I G H T G A I N B E T W E E N 14 A N D 28 D A Y S O F A G E WERE E X P R E S S E D A S P E R C E N T A G E S O F T H E A M O U N T S P R E S E N T I N T H E O R G A N S A T 14 D A Y S ( B E F O R E A R G I N I N E S U P P L E M E N T A T I O N ) . T H E S E C O M P A R I S O N S A R E I L L U S T R A T E D I N F l G . 11 A N D 12. A C C U M U L A T I O N O F M U S C L E A N D L I V E R DNA, RNA A N D P R O T E I N C O N S T I T U E N T S I N C R E A S E D W I T H A R G I N I N E S U P P L E M E N T A T I O N TO T H E H I G H L Y S I N E D I E T S C O M P A R E D W I T H T H E L Y S I N E A D E Q U A T E D I E T (0.86$). R A T E 87 160 _ 150 -=t z - 140 u H O o: Q_ < _ J u 130 120 110 100 F I G . 9 D I E T A R Y T R E A T M E N T 0.86$ L Y S I N E 1 .36$ L Y S I NE 1 + 0.013$ L-ARG. L Y S I N E + 0.293$ L-ARG . z Q <t z an L U > < U J on - 80 - 70 - 60 50 I M P R O V E M E N T I N M U S C L E P R O T E I N / D N A A N D RNA/DNA R A T I O S F R O M C H I C K S F E D E X C E S S L Y S I N E W I T H A R G I N I N E . A R G I N I N E WAS A D D E D TO D I E T S C O N T A I N I N G 1.36, 1.86$ L Y S I N E A F T E R TWO W E E K S O F F E E D I N G . P R O T E I N / D N A , RNA/DNA R A T I O S A T F O U R W E E K S O F A G E A R E E X P R E S S E D A S P E R C E N T A G E S O F T H E R A T I O S A T TWO W E E K S . 88 D I E T A R Y T R E A T M E N T 1. b L Y S I N E '0 L Y S I N E + 0.013$ L-ARG. >o L Y S I N E + 0.293$ L^ARG . H O sr h-< _i 130-120-110-100-90" 80 _ F I G . 10. <t Q <£ Z u > < 130 - 120 - 110 100 • 90 80 < < < < >< C H A N G E S I N L I V E R P R O T E I N / D N A A N D RNA/DNA R A T I O S F R O M C H I C K S F E D E X C E S S L Y S I N E W I T H A R G I N I N E . A R G I N I N E WAS A D D E D TO D I E T S C O N T A I N I N G 1.36, 1.86$ L Y S I N E A F T E R TWO W E E K S O F F E E D I N G . P R O T E I N / D N A , RNA/DNA R A T I O S A T F O U R W E E K S O F A G E A R E E X P R E S S E D A S P E R C E N T A G E S O F T H E R A T I O S A T TWO W E E K S . 89 A B C A B C A B C LYSINE (fo) 0.86 1.36 1.86 ARGININE {%) 0.013 0.293 FIG. 11. EFFECTS OF SUPPLEMENTING HIGH LYSINE DIETS WITH ARGININE ON ACCUMULATION OF GASTROCNEMIUS MUSCLE DNA (A), RNA (B) AND PROTEIN (c). THE AMOUNTS OF MUSCLE CONSTITUENTS ACCUMULATED FROM 14-TH TO 2 8 T H DAY OF FEEDING ARE EXPRESSED AS PERCENTAGES OF THE AMOUNTS AT 14TH DAY OF FEED I NG . 300 -A B C D A B C D A B C D (^LYSINE {%) 0.86 1.36 1.86 ARGININE {%) 0.013 0.293 FIG. 12. EFFECTS OF SUPPLEMENTING HIGH LYSINE DIETS WITH ARGININE ON ACCUMULATION OF LIVER DNA (A), RNA (B), PROTEIN ( c ) AND BODY WEIGHT G A I N / C H I C K (D). T H E AMOUNTS OF LIVER CONSTITUENTS ACCUMULATED AND BODY WEIGHT GAIN FROM 14 TO 28TH DAY OF FEEDING ARE EXPRESSED AS PERCENTAGES OF THE AMOUNT AT 14TH DAY OF FEEDING. O F G R O W T H ( F l G . 12) I M P R O V E D W I T H A H I G H E R L E V E L O F A R G I N I N E S U P P L E -M E N T A T I O N (0.293$) TO E X C E S S L Y S I N E D I E T (1.86$). THE R A T E O F P R O T E I N A C C U M U L A T I O N WAS G R E A T E R I N B O T H T H E M U S C L E A N D T H E L I V E R T H A N DNA A C C U M U L A T I O N W H E N A R G I N I N E WAS S U P P L E M E N T E D T O E X C E S S L Y S I N E D I E T S OR I N C H I C K S R E C E I V I N G A N A D E Q U A T E L YS I NE .D H>E T. R E S U L T S O F T H E P R E S E N T E X P E R I M E N T C L E A R L Y D E M O N S T R A T E D T H A T DNA, RNA A N D P R O T E I N A C C U M U L A T I O N W E R E I M P A I R E D I N T H E M U S C L E A N D L I V E R O F C H I C K S W H E N E X C E S S L Y S I N E WAS F E D , A N D A R G I N I N E S U P P L E M E N T A T I O N E L I M I N A T E D T H E I M P A I R M E N T . R E S U L T S I N D I C A T E T H A T A R G I N I N E B E C O M E S L I M I T I N G F O R C E L L U L A R G R O W T H ( H Y P E R P L A S I A A N D H Y P E R T O P H Y ) W H E N E X C E S S A M O U N T S O F L Y S I N E A R E P R E S E N T I N T H E C H I C K D I E T . I T WAS D E M O N S T R A T E D ( T R I A L 1) T H A T B O D Y A N D T I S S U E G R O W T H W E R E I N P R O P O R T I O N TO A V A I L A B L E D I E T A R Y L Y S I N E , T H E H I G H L E V E L O F L Y S I N E C A U S I N G D E P R E S S E D F E E D I N T A K E A N D P O O R G R O W T H . I T WAS A L S O D E M O N S T R A T E D ( S M I T H , 1968J A U S T I C A N D N E S H E I M , 1970) I N C H I C K N U T R I T I O N T H A T T H E A D V E R S E E F F E C T S S U C H A S R E D U C E D F E E D I N T A K E A N D G R O W T H D E P R E S S I O N R E S U L T I N G F R O M E X C E S S D I E T A R Y L Y S I N E WERE A L L E V I A T E D B Y S U P P L E M E N T A R Y A R G I N I N E . S U M M A R Y A N D C O N C L U S I O N THE E F F E C T S O F E X C E S S D I E T A R Y L Y S I N E F O L L O W E D B Y A R G I N I N E S U P P L E M E N T A T I O N ON DNA, RNA A N D P R O T E I N A C C U M U L A T I O N I N C H I C K G A S T R O C N E M I U S M U S C L E A N D L I V E R W E R E D E T E R M I N E D . C H I C K S W E R E F E D 0.86, 1.36 A N D 1.86$ L Y S I N E D I E T S F O R 14 D A Y S ( S U P P L E M E N T A L L Y S I N E O F 0.5, 1.0 A N D 1.5$ R E S P E C T I V E L Y ) . A R G I N I N E WAS T H E N A D D E D F R O M 14 TO 28 D A Y S TO D I E T S C O N T A I N I N G 1.36 A N D 1.86$ L Y S I N E . C E L L U L A R P R O T E I N S A N D RNA I N T H E M U S C L E A N D L I V E R W E R E D E P R E S S E D B Y E X C E S S L Y S I N E A N D I M P R O V E D M A R K E D L Y W H E N A R G I N I N E WAS A D D E D TO D I E T S C O N T A I N I N G E X C E S S L Y S I N E . C H I C K G R O W T H A N D A C C U M U L A T E D M U S C L E A N D L I V E R DNA, RNA A N D P R O T E I N I N C R E A S E D A T A F A S T E R R A T E I N C H I C K S F E D A R G I N I N E S U P P L E M E N T E D D I E T S T H A N W I T H T H E A D E Q U A T E L Y S I N E (0.86$) D I E T . T H E R E S U L T S D E M O N S T R A T E D T H A T W H E N E X C E S S L Y S I N E I S P R E S E N T I N T H E C H I C K D I E T A R G I N I N E B E C O M E S L I M I T I N G F O R M U S C L E A N D L I V E R C E L L U L A R G R O W T H . 93 TRIAL 3. T H I S S T U D Y WAS D E S I G N E D TO D E T E R M I N E T H E R E L A T I V E E F F E C T S O F D I E T A R Y P R O T E I N Q U A L I T Y A N D Q U A N T I T Y ON T H E A C C U M U L A T I O N O F G A S T R O C N E M I U S M U S C L E A N D L I V E R DNA, RNA A N D P R O T E I N . A V E R A G E B O D Y W E I G H T G A I N A N D F E E D I N T A K E O F C H I C K S F E D D I F F E R E N T D I E T A R Y P R O T E I N S F O R 14 D A Y S A R E P R E S E N T E D I N T A B L E 14 A N D F O R 28 D A Y S I N T A B L E 15. C H I C K S C O N S U M I N G T H E 20$ D I E T A R Y P R O T E I N A T E M ORE F E E D (P -= 0.01 ) A N D G A I N E D W E I G H T A T A M ORE R A P I D R A T E (P "=0.01) T H A N C H I C K S F E D L O W E R D I E T A R Y P R O T E I N T H R O U G H O U T T H E S T U D Y . T H E S E R E S U L T S I N D I C A T E T H A T 20$ P R O T E I N I S R E Q U I R E D F O R O P T I M U M C H I C K G R O W T H D U R I N G T H I S P E R I O D . R E S U L T S A G R E E W I T H T H E P R O T E I N R E Q U I R E M E N T F O R C H I C K G R O W T H U P TO S I X W E E K S O F A G E T H A T H A S B E E N E S T A B L I S H E D A T 20$ O F T H E D I E T F O R R E P L A C E M E N T P U L L E T S A N D 23$ F O R B R O I L E R S (NRC, 1971). H O W E V E R , B O D Y W E I G H T G A I N S I N T H E W H E A T - S O Y A A N D W H E AT-0.5$ L Y S I N E G R O U P S (15$ P R O T E I N ) D I D NOT D I F F E R F R O M E A C H O T H E R S I G N I F I C A N T L Y . W H E A T A S A S O L E S O U R C E O F P R O T E I N S U P P O R T E D T H E L O W E S T G R O W T H R A T E . G A S T R O C N E M I U S M U S C L E W E I G H T , DNA, RNA, A N D P R O T E I N C O N T E N T I N C H I C K S F E D D I F F E R E N T P R O T E I N S A R E I L L U S T R A T E D I N T A B L E S 12 A N D 13. THE M U S C L E W E I G H T A N D B I O C H E M I C A L C O M P O N E N T S W E R E I N G R E A T E S T A M O U N T A T 14 D A Y S (P <0.05) A N D 28 D A Y S (P <=0.01) O F A G E I N C H I C K S F E D 20$ P R O T E I N A N D M I N I M U M I N T H E W H E A T F E D G R O U P . H O W E V E R , T H E P A R A M E T E R S I N C H I C K S F E D W H E A T - S O Y A A N D W H E AT-0.5$ L - L Y S I N E (15$ P R O T E I N ) D I D N O T D I F F E R A T 28 D A Y S O F A G E . H O W A R T H (1972) S H O W E D T H A T DNA, RNA, P R O T E I N C O N T E N T I N T H E M U S C L E WERE L E S S I N R A T S F E D 6, 12 A N D 18$ D I E T A R Y P R O T E I N C O M P A R E D TO T H O S E R E C E I V I N G 24$ D I E T A R Y P R O T E I N . L I V E R W E I G H T A N D L I V E R DNA, RNA A N D P R O T E I N C O N T E N T F R O M C H I C K S TABLE 12.--DNA, RNA A N D P R O T E I N C O N T E N T I N T H E G A S T R O C N E M I U S M U S C L E O F C H I C K S F E D D I F F E R E N T L E V E L S A N D Q U A L I T Y OF D I E T A R Y P R O T E I N F O R TWO W E E K S D 1 E T A R Y M U S C L E WT. DNA RNA P R O T E I N T R E A T M E N T MG. 2"* MG . 1 WHEAT B A S A L (15$ P R O T E I N ) 682.9A 0.36 A 0.94 A A * * 70.3 WHEAT B A S A L + 0.5$ L Y S I N E 1271.8 B 0.68 B C 2.30 B 180.4 B W H E A T - S O Y A (15$ P R O T E I N ) 955.1 C 0.41 B 1 .52 A 149.0 B W H E A T - S O Y A - F I S H M E A L (20$ P R O T E I N ) 1727.6 D 0.86 C 3.23° 263.2° * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P < 0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P "=0.01). WHEAT B A S A L C O N T A I N E D 0.36$ L Y S I N E . DATA R E P R E S E N T T H E T O T A L O B T A I N E D F R O M T H E R I G H T A N D L E F T WET G A S T R O C N E M I U S M U S C L E P E R C H I C K S A M P L E D . TABLE 13.--DNA, RNA AND PROTEIN CONTENT IN THE GASTROCNEMIUS MUSCLE OF CHICKS FED DIFFERENT LEVELS AND QUALITY OF DIETARY PROTEIN FOR FOUR WEEKS DIETARY MUSCLE WT. DNA RNA PROTEIN TREATMENT MG . MG . WHEAT BASAL (15$ PROTEIN) 101 8.2 A A* 0.69 1 .20A 149.9 A WHEAT BASAL + 0.5$ LYSINE 4547.5 B 2.40 B 5.12 8 812.7 8 WHEAT-SOYA (15$ PROTEIN) 4546.2 B 2.20 B 5.78 B 796.8 B WHEAT-SOYA-FISHMEAL (20$ PROTEIN) 7582.63 C 3.30° 11.36 C 1685.7 C *VALUES FOLLOWED BY A DIFFERENT SUPERSCR1PT ARE SIGNIFICANTLY DIFFERENT (P -= 0.05). **VALUES FOLLOWED BY A DIFFERENT SUPERSCR1PT ARE SIGNIFICANTLY DIFFERENT (P < 0.01). WHEAT BASAL CONTAINED 0.36$ LYSINE. DATA REPRESENT THE TOTAL OBTAINED FROM THE RIGHT AND LEFT WET GASTROCNEMIUS MUSCLE PER CHICK SAMPLED . TABLE 14.--BODY W E I G H T G A I N , F E E D I N T A K E , L I V E R DNA, RNA A N D P R O T E I N C O N T E N T I N C H I C K S F E D D I F F E R E N T L E V E L S A N D Q U A L I T Y OF D I E T A R Y P R O T E I N F O R TWO W E E K S D I E T A R Y WT. G A I N / C H I C K F E E D 1 N T A K E / C H 1 C K L I V E R WT. DNA RNA P R O T E I N T R E A T M E N T G . * * G.* MG . / L I V E R * WHEAT B A S A L (15$ P R O T E I N ) 2 6 A 92 A 1.4 A 2.7 A 10. 8 A 210.0 A WHEAT B A S A L + 0.5$ L - L Y S I N E 96 B 21 1 B 4.1 B 10.7 B C 39.0 B 677.9 B C W H E A T - S O Y A (15$ P R O T E I N ) 8 3 B 193 B 3.1 C 8.0 B 29.1 C 581 .1 B W H E A T - S O Y A - F I S H M E A L 38.0 B (20$ P R O T E I N ) 150° 252° 4.9° 12.3 C 764.8° * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R 1 P T A R E S I G N I F I C A N T L Y D 1 F F E R E N T (P ' «= 0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R 1 P T A R E S I G N I F l C A N T L Y D I F F E R E N T (P < 0.01). WHEAT B A S A L C O N T A I N E D 0.36$ L Y S I N E . TABLE 15.--BODY W E I G H T G A I N , F E E D I N T A K E , L I V E R DNA, RNA A N D P R O T E I N I N C H I C K S F E D D I F F E R E N T L E V E L S A N D Q U A L I T Y O F D I E T A R Y P R O T E I N F O R F O U R W E E K S D I E T A R Y WT. G A i N / C H I C K F E E D I N T A K E / C H I C K L I V E R WT. DNA RNA P R O T E I N T R E A T M E N T G.** G.* : : G.* MG . / L I V E R * WHEAT B A S A L (15$ P R O T E I N ) 56A 21 7A 2.9A 7.2A 18.5A 398.5A WHEAT B A S A L + 0.5$ L - L Y S I N E 276B 704B 12.1B 26.7B 85.2BC 1749.68 W H E A T - S O Y A (15$ P R O T E I N ) 249B 647° 10.oB 24.1 8 69.7B 1437.5B W H E A T - S O Y A - F I S H M E A L (20$ P R O T E I N ) 487° . 896° 15.4° 32.3B 101 .5° 2345.3° * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R 1 P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P *= 0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R 1 P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P *=0.01 ). WHEAT B A S A L C O N T A I N E D 0.36$ L Y S I N E . to F E D D I F F E R E N T D I E T A R Y P R O T E I N S F O R 14 A N D 28 D A Y S A R E R E S P E C T I V E L Y S H O W N I N TABLES.14 A N D 15. L I V E R W E I G H T S O F C H I C K S F E D W H E A T A S A S O L E S O U R C E O F P R O T E I N , WHEAT-0.5$ L Y S I N E A N D W H E A T - S O Y A D I E T S W E R E S I G N I F I C A N T L Y (P -=0.05) L E S S T H A N F O R T H E 20$ P R O T E I N D I E T . L L V E R DNA, RNA A N D P R O T E I N W E R E S I G N I F I C A N T L Y (P «= 0.05) L O W E R I N T H E W H E A T (15$ D I E T A R Y P R O T E I N ) F E D C H I C K S T H A N I N B I R D S R E C E I V I N G 20$ P R O T E I N , W H E A T - S O Y A (15$ P R O T E I N ) A N D WHEAT-0.5$ L Y S I N E D I E T S . THE L I V E R B I O C H E M I C A L C O N S T I T U E N T S (DNA, RNA, P R O T E I N ) I N T H E WHEAT-0.5$ L Y S I N E A N D W H E A T - S O Y A G R O U P S D I D NOT D I F F E R T H R O U G H O U T T H E S T U D Y ( T A B L E S 14 A N D 15). L I V E R DNA A N D RNA C O N T E N T I N C H I C K S F E D T H E 20$ P R O T E I N WAS S L I G H T L Y H I G H E R T H A N T H A T O F C H I C K S C O N S U M I N G T H E WHEAT-0.5$ L Y S I N E D I E T D U R I N G B O T H P E R I O D S O F D E T E R M I N A T I O N . L L V E R P R O T E I N A T 14 D A Y S O F A G E D I D NOT D I F F E R B E T W E E N T H E S E G R O U P S O F C H I C K S B U T T H E C H I C K S F E D T H E 20$ P R O T E I N1-D I E T HAD S I G N I F I C A N T L Y (P «= 0. 05) MORE L I V E R P R O T E I N A T 28 D A Y S T H A N T H E WHEAT-0.5$ L Y S I N E F E D G R O U P . P R E V I O U S R E P O R T S ( A L L I S O N _ET _AL . 1963) H A V E I N D I C A T E D T H A T R A T L I V E R RNA A N D P R O T E I N I N C R E A S E D W I T H I N C R E A S I N G N I T R O G E N I N T A K E OR N U T R I T I V E V A L U E O F D I E T A R Y P R O T E I N . THE G R E A T E S T L I V E R RNA A N D P R O T E I N WAS O B S E R V E D I N R A T S F E D E G G P R O T E I N A N D M I N I M U M V A L U E S I N T H O S E F E D W H E A T G L U T E N . B A N K S _ E T _ A L . (1 964) A L S O D E M O N S T R A T E D T H A T S U P P L E M E N T A T I O N O F W H E A T G L U T E N W I T H C A S E I N I N C R E A S E D C E L L U L A R RNA A N D P R O T E I N C O N T E N T . R E S U L T S W I T H C H I C K S F E D D I F F E R E N T Q U A N T I T Y A N D Q U A L I T Y O F D I E T A R Y P R O T E I N I N T H E P R E S E N T T R I A L A G R E E W I T H R E S U L T S R E P O R T E D I N R A T S . T O T A L L I V E R DNA C O N T E N T R E F L E C T E D C H A N G E S I N L I V E R W E I G H T I N P R O P O R T I O N TO B O D Y G R O W T H A L S O N O T E D I N T R I A L 1. C A M P B E L L A N D K O S T E R L I T Z (1950) R E P O R T E D T H A T L I V E R DNA I N C R E A S E D A S B O D Y W E I G H T I NC RE A S E D . G A S T R O C N E M I U S M U S C L E C E L L S I Z E , M E A S U R E D B Y P R O T E I N / D N A A N D RNA/DNA I N C R E A S E D M A R K E D L Y I N T H E C H I C K A T 14 D A Y S ( F I G . 13) A N D 28 D A Y S ( F I G . 14) O F A G E W I T H I N C R E A S E D D I E T A R Y P R O T E I N . THE R E S U L T A G R E E S W I T H A N D C O N F I R M S T H E S A M E O B S E R V A T I O N M A D E I N T R I A L 1 A S T H E L E V E L O F L Y S I N E I N T H E C H I C K D I E T I N C R E A S E D F R O M 0.36 TO 0.61 A N D 0.86$. THE I N C R E A S E WAS G R E A T E R A T 28 D A Y S T H A N A T 14 D A Y S O F A G E . R E S U L T S I N D I C A T E T H A T P R O T E I N WAS B E I N G S Y N T H E S I Z E D A N D A C C U M U L A T E D I N T H E M U S C L E I N A G R E A T E R A M O U N T A T 28 D A Y S T H A N A T 14 D A Y S O F A G E . T H I S V I E W I S I N A G R E E M E N T W I T H R E P O R T S T H A T P O S T - H A T C H I N G OR P O S T -N A T A L G R O W T H O F M U S C L E S I N B I R D S OR M A M M A L S WAS D U E TO A N I N C R E A S E I N C E L L S I Z E S I N C E D I F F E R E N T I A T I O N ( C E L L S P E C I A L I Z A T I O N ) O F S K E L E T A L M U S C L E T I S S U E WAS C O M P L E T E D P R I O R TO H A T C H I N G O F C H I C K S ( S M I T H , 1963) OR B I R T H O F M A M M A L S (MACCALLUM, 1898j MCMEEKAN, 1940-41; E L I O T E T A L . 1943; A N D M E A R A , 1947). M O R E O V E R , T H E I N C R E A S E I N P R O T E I N / D N A A N D RNA/DNA R A T I O S R E F L E C T E D G R O W T H R E S P O N S E I N T H E C H I C K TO D I E T A R Y P R O T E I N S ( T A B L E S 14 A N D 15). M O N T G O M E R Y _ET _ A L . (1964) R E P O R T E D T H A T M U S C L E S WERE T H E C O R R E C T W E I G H T F O R T H E B O D Y . MEHNER (1938) A L S O O B S E R V E D A C L O S E A S S O C I A T I O N B E T W E E N M U S C L E F I B E R S I Z E A N D B O D Y S I Z E A T V A R I O U S A G E S I N C H I C K E N S . S I M I L A R O B S E R V A T I O N S WERE M A D E I N T H E R A B B I T , S W I N E , S H E E P A N D C A T T L E ( M C M E E K A N , 1940-41; J O U B E R T , 1956). M O S S _E_T _AL, (1 964) A L S O O B S E R V E D T H A T T H E W E I G H T O F M U S C L E / N UC L E U S ( C Y T O P L A S M / N U C L E U S ) I N C R E A S E D T H R E E TO F O U R F O L D I N T H E C H I C K B E T W E E N O N E A N D 28 D A Y S O F A G E . L I V E R P R O T E I N / D N A A N D RNA/DNA R A T I O S A T 14 D A Y S O F A G E S H O W E D A N 100 A B C D . A B C D FIG. 13. EFFECTS OF THE QUALITY AND QUANTITY OF DIETARY PROTEIN ON GASTROCNEMIUS MUSCLE PROTEIN/DNA AND RNA/DNA RATIOS IN THE CHICK AT TWO WEEKS OF AGE. A, B, C, D RESPECTIVELY REPRESENTS WHEAT-BASAL, WHEAT-BASAL PLUS 0.5$ L-LYSINE, WHEAT-SOYA (15$ PROTEIN) AND 20$ PROTEIN DIET. 101 700 • 600 • A B C D A B C D FIG. 14. EFFECTS OF THE QUALITY AND QUANTITY OF DIETARY PROTEIN ON GASTROCNEMIUS MUSCLE PROTEIN/DNA AND RNA/DNA RATIOS IN THE CHICK AT FOUR WEEKS OF AGE. A, B, C, D RESPECTIVELY REPRESENTS WHEAT-BASAL, WHEAT-BASAL PLUS 0.5$ L-LYSINE, WHEAT-SOYA (15$ PROTEIN) AND 20$ PROTEIN DIET. O P P O S I T E P A T T E R N TO T H E M U S C L E (F I G. 15 C O M P A R E D W I T H F L G . 13). THE R A T I O S D E C R E A S E D I N T H E L I V E R A N D I N C R E A S E D I N T H E M U S C L E W I T H I N C R E A S E D P R O T E I N L E V E L OR N U T R I T I V E V A L U E O F T H E D I E T A R Y P R O T E I N . L I V E R G R O W T H WAS S H O W N I N T R I A L 1 TO B E B Y A N I N C R E A S E I N C E L L N U M B E R ( H Y P E R P L A S I A ) I N C H I C K S A T 14 D A Y S O F A G E . THE D E C R E A S E I N L I V E R P R O T E I N / D N A A N D RNA/DNA R A T I O S WAS T H E R E F O R E A C C O U N T E D F O R B Y A C C U M U L A T I O N O F DNA (A R A P I D S T A T E O F M U L T I P L I C A T I O N O F L I V E R C E L L S ) W I T H L I T T L E OR NO I N C R E A S E I N P R O T E I N A N D RNA. M U S C L E G R O W T H O F C H I C K S A T 14 D A Y S O F A G E , ON T H E O T H E R H A N D , WAS B Y A N I N C R E A S E I N C E L L U L A R P R O T E I N S A N D RNA A N D N O T I N C E L L U L A R DNA C O N T E N T , T H E N U M B E R O F C E L L S OR F I B R E S ( T R I A L 1). THE I N C R E A S E I N C E L L U L A R P R O T E I N S A N D RNA A N D T H E R E L A T I V E C O N S T A N C Y O F C E L L N U M B E R WAS E N O U G H TO C A U S E T H E I N C R E A S E S I N P R O T E I N / D N A A N D RNA/DNA R A T I O S I N T H E M U S C L E A S T H E Q U A N T I T Y A N D N U T R I T I V E V A L U E O F D I E T A R Y P R O T E I N S I M P R O V E D . M O R E O V E R , T H E S E R E S U L T S S U G G E S T E D T H A T P R O T E I N I N T A K E WAS U S E D F O R M U S C L E G R O W T H W H I C H A C C O U N T E D F O R T H E I N C R E A S E I N B O D Y W E I G H T . L I V E R C E L L S I Z E , I N D I C A T E D B Y P R O T E I N / D N A A N D RNA/DNA, I N C R E A S E D A S I N T H E M U S C L E W I T H I N C R E A S E D D I E T A R Y P R O T E I N I N T H E C H I C K A T 28 D A Y S O F A G E ( F I G . 14 A N D 16). THE I N C R E A S E WAS G R E A T E R I N T H E L I V E R P R O T E I N / D N A R A T I O T H A N I N RNA/DNA R A T I O I N D I C A T I N G A N A C T I V E L I V E R P R O T E I N S Y N T H E S I S A N D A C C U M U L A T I O N A S T H E Q U A N T I T Y A N D Q U A L I T Y O F D I E T A R Y P R O T E I N I M P R O V E D . THE D E C R E A S E I N L I V E R P R O T E I N / D N A A N D RNA/DNA R A T I O S A T 14 D A Y S F O L L O W E D B Y A N I N C R E A S E I N T H E R A T I O S A T 28 D A Y S O F A G E W I T H H I G H E R L E V E L S O F D I E T A R Y P R O T E I N WAS A L S O O B S E R V E D W I T H A D D E D D I E T A R Y L Y S I N E I N T R I A L 1. R E S U L T S O F T R I A L S 1 A N D 3 ON C E L L S I Z E (PROTEiN/DNA, RNA/DNA)AT 14 A N D 28 D A Y S O F A G E S T R O N G L Y A B C D A B C D G. 15. EFFECTS OF THE QUALITY AND QUANTITY OF DIETARY PROTEIN ON LIVER PROTEIN/DNA AND RNA/DNA RATIOS IN THE CHICK AT TWO WEEKS OF AGE. A, B, C, D RESPECTIVELY REPRESENTS WHEAT-BASAL, WHEAT-BASAL PLUS 0.5$ L-LYSINE, WHEAT-SOYA (15$ PROTEIN) AND 20$ PROTEIN DIET. 104 FIG. 16. EFFECTS OF THE QUALITY AND QUANTITY OF DIETARY PROTEIN ON LIVER PROTEIN/DNA AND RNA/DNA RATIOS IN THE CHICK AT FOUR WEEKS OF AGE. A, B, C, D RESPECTIVELY REPRESENTS WHEAT-BASAL, WHEAT-BASAL PLUS 0.5$ L-LYSINE, WHEAT-SOYA (15$ PROTEIN) AND 20$ PROTEIN DIET. 105 I N D I C A T E T H A T L I V E R G R O W T H I S B Y A N I N C R E A S E I N C E L L N U M B E R ( H Y P E R P L A S I A ) D U R I N G T H E F I R S T 14 D A Y S O F A G E A N D B Y I N C R E A S E S B O T H I N C E L L N U M B E R A N D I N C E L L S I Z E ( H Y P E R T R O P H Y ) F R O M 14 TO 28 D A Y S O F A G E . E N E S C O A N D L E B L O N D (1962) R E P O R T E D T H A T D U R I N G T H E I N I T I A L 17 D A Y S O F P O S T - N A T A L L I F E , T H E L I V E R O F T H E R A T G R E W B Y A N I N C R E A S E I N C E L L N U M B E R A N D W I T H O N L Y T R I V I A L C H A N G E S I N C E L L S I Z E . F R O M 17 TO 34 OR 48 D A Y S , H O W E V E R , T H E I N T E N S I T Y O F C E L L M U L T I P L I C A T I O N S L O W E D DOWN A N D S O M E I N C R E A S E S I N W E I G H T D U E TO I N C R E A S E I N C E L L S I Z E B E G A N TO A P P E A R . THE A B S O L U T E A M O U N T S O F DNA, RNA A N D P R O T E I N A N D B O D Y W E I G H T G A I N A C C U M U L A T E D F R O M 14 TO 28 D A Y S O F A G E W E R E E S T I M A T E D B Y T H E D I F F E R E N C E S B E T W E E N T H E V A L U E S A T 14 A N D 28 D A Y S . A C C U M U L A T I O N O F T H E S E L I V E R A N D M U S C L E C O N S T I T U E N T S I N C H I C K S ON T H E W H E A T - B A S A L , W H E A T -0.5$ L Y S I N E , W H E A T - S O Y A A N D 20$ P R O T E I N ( W H E A T - S 0 Y B E A N P L U S F I S H M E A L ) D I E T S WAS E X P R E S S E D R E L A T I V E TO T H E A M O U N T S A C C U M U L A T E D ON T H E 20$ P R O T E I N D I E T I N O R D E R TO P R O V I D E A C O M M O N B A S I S F O R C O M P A R I N G T H E E F F E C T S O F P R O T E I N Q U A L I T Y A N D Q U A N T I T Y ON DNA, RNA A N D P R O T E I N A C C U M U L A T I O N ( F L G . 17 A N D 18). A C C U M U L A T I O N O F T H E T H R E E M U S C L E A N D L I V E R C O N S T I T U E N T S A N D B O D Y W E I G H T G A I N P R O G R E S S I V E L Y D E C R E A S E D A S T H E D I E T A R Y S T R E S S I N C R E A S E D F R O M A LOW P R O T E I N Q U A L I T Y ( W H E A T - S O Y A A N D WHEAT-0.5$ L Y S I N E D I E T S ) T O A P O O R Q U A L I T Y ( T H E W H E A T D I E T ) . B O D Y W E I G H T G A I N ( F L G . 18) WAS D E P R E S S E D M O S T S E V E R E L Y I N E A C H O F T H E LOW Q U A L I T Y P R O T E I N D I E T S . P R O T E I N A C C U M U L A T I O N I N T H E M U S C L E ( F I G . 17) A N D L I V E R ( F I G . 18) WAS D E P R E S S E D M O R E S E V E R E L Y T H A N DNA A C C U M U L A T I O N . G R E A T E R D E P R E S S I O N I N T I S S U E P R O T E I N T H A N DNA A C C U M U L A T I O N I S C O N S I S T E N T W I T H I N C R E A S I N G 106 2 O I -< _l O U «4 < & PROTEIN WHEAT-SOYA WHEAT-BASAL + 0.5$ LYSINE B C WHEAT-BASAL FIG. 17. EFFECTS OF DIETARY PROTEIN LEVEL AND QUALITY ON RELATIVE ACCUMULATION OF DNA (A), RNA (B) AND PROTEIN (C) IN THE CHICK GASTROCNEMIUS MUSCLE BETWEEN 14 AND 28 DAYS OF AGE. 107 z o 3 D o o <I UJ > f— < 100 80 60 40 20. 0L / ** IX XX XX XK *X X* )<|c XX xx / / / / / / / / / XX XX R / / / / /:• / / 1 +Y. IKK tx XX PI /" c D FIG. 1 A B C i PROTE D I N B C WHEAT-SOYA WHEAT-BASAL WHEAT-BASAL + 0.5$ L-LYS I NE EFFECTS OF DIETARY PROTEIN LEVEL AND QUALITY ON RELATIVE ACCUMULATION OF DNA (A), RNA (B), PROTEIN (C) IN CHICK LIVER AND BODY WEIGHT G A I N / C H I C K (D) BETWEEN 14 AND 28 DAYS OF AGE. P R O T E I N / D N A R A T I O W I T H I M P R O V E D N U T R I T I V E V A L U E O F D I E T A R Y P R O T E I N A T 28 D A Y S O F A G E . RNA A C C U M U L A T I O N I N B O T H O R G A N S WAS I N T E R M E D I A T E B E T W E E N DNA A N D P R O T E I N A C C U M U L A T I O N I R R E S P E C T I V E O F O I E T A R Y P R O T E I N Q U A L I T Y OR Q U A N T I T Y . R E S U L T S T H A T L Y S I N E A N D P R O T E I N D E F I C I E N C Y D E P R E S S E D P R O T E I N A C C U M U L A T I O N MORE S E V E R E L Y T H A N DNA A C C U M U L A T I O N I N T H E M U S C L E A N D L I V E R O F T H E C H I C K S U G G E S T T H A T A L T H O U G H T I S S U E C E L L S I Z E M A Y B E G R E A T L Y R E D U C E D I N C H I C K S U N D E R N O U R I S H E D F R O M A N E A R L Y A G E , D E G E N E R A T I O N OR L O S S O F C E L L S OR F I B R E S D O E S NOT O C C U R . M O N T G O M E R Y (1962A) O B S E R V E D I N J A M A I C A N I N F A N T S D Y I N G O F S E V E R E P R O T E I N M A L N U T R I T I O N T H A T S O M E F I B R E S I N T H E S A R T O R I U S M U S C L E S D I S A P P E A R E D , T H E D I A M E T E R O F T H E R E M A I N D E R WAS G R E A T L Y R E D U C E D A N D T H E R E WAS A R E L A T I V E I N C R E A S E I N I N T E R S T I T I A L C O N N E C T I V E T I S S U E . H O W E V E R , M O N T G O M E R Y _ET _ A L . (1964) W E R E U N A B L E TO D E M O N S T R A T E S U C H C H A N G E S I N T H E S A R T O R I U S M U S C L E S O F U N D E R N O U R I S H E D B I R D S . T H E O B S E R V A T I O N I N T H E P R E S E N T T R I A L T H A T L Y S I N E A N D P R O T E I N D E F I C I E N C Y D E P R E S S E D P R O T E I N A C C U M U L A T I O N M O R E S E V E R E L Y T H A N DNA A C C U M U L A T I O N I N T H E C H I C K I S I N A G R E E M E N T W I T H M O N T G O M E R Y _E_T _ A L . (1964) A N D M A Y E X P L A I N WHY C H I C K S U N D E R S E V E R E P R O T E I N A N D / O R C A L O R I E S M A L N U T R I T I O N C A N GROW TO A N O R M A L S I Z E W H E N R E H A B I L I T A T E D ON D I E T S A D E Q U A T E I N P R O T E I N A N D C A L O R I E S . A P R E V I O U S R E P O R T ( M E N D E S A N D W A T E R L O W , 1958) I N D I C A T E D A G R O S S R E D U C T I O N I N B O D Y W E I G H T , P R O T E I N A N D DNA I N T H E M U S C L E A N D L I V E R O F R A T S F E D A P R O T E I N D E F I C I E N T D I E T . IT WAS A L S O O B S E R V E D T H A T A P R O T E I N D E F I C I E N T D I E T P R I M A R I L Y R E D U C E D M U S C L E P R O T E I N S Y N T H E S I S W H I L E A C A L O R I E D E F I C I E N C Y I N T E R R U P T E D M U S C L E C E L L G R O W T H ( C H E E K A N D H I L L , 1970; H I L L ^ T _ A L . 1970). HOWEVER, HOWARTH (1972) S H O W E D T H A T P R O T E I N S Y N T H E S I S HAD P R I O R I T Y O V E R DNA S Y N T H E S I S W H E N M U S C L E G R O W T H WAS I M P A I R E D B Y F E E D I N G A P R O T E I N D E F I C I E N T D I E T TO R A T S . L I V E R P R O T E I N A N D N I T R O G E N B A L A N C E O F R A T S W E R E I M P R O V E D B Y A D D I N G C A R B O H Y D R A T E OR F A T TO A S T A N D A R D D I E T A D E Q U A T E I N P R O T E I N TO I N C R E A S E E N E R G Y I N T A K E . D U R A N D A N D P E R I O T (1969), H O W E V E R , D E M O N S T R A T E D W I T H O N E - Y E A R - O L D R A T S T H A T D E C R E A S I N G L I P I D S A N D C A R B O H Y D R A T E S I N A 13$ P R O T E I N D I E T W H I C H D E C R E A S E D E N E R G Y I N T A K E C A U S E D A R E D U C T I O N I N T O T A L L I V E R DNA, P R O T E I N A N D RNA B Y 13, 12 A N D 17$ R E S P E C T I V E L Y . T H E M U S C L E S H O W E D L I T T L E E F F E C T A S A R E S U L T O F T H E E N E R G Y D E F I C I E N C Y . THE C O M P L E X I T I E S F U R T H E R I N C R E A S E T H E D I F F I C U L T Y I N A T T E M P T I N G TO D I S T I N G U I S H B E T W E E N P R O T E I N A N D C A L O R I E D E F I C I E N C I E S ON T I S S U E G R O W T H . IN T H E P R E S E N T T R I A L M . E . : P R O T E I N R A T I O I N T H E 20$ P R O T E I N D I E T WAS 67 W H I L E T H E V A L U E I N T H E LOW P R O T E I N G R O U P S WAS 85. D I E T A R Y P R O T E I N Q U A L I T Y R A T H E R T H A N E N E R G Y WAS C O N S I D E R E D T H E P R I M A R Y F A C T O R A S S H O W N B Y I N C R E A S E D F E E D C O N S U M P T I O N A N D G R O W T H ( T A B L E S 14 A N D 15) F O L L O W I N G T H E A D D I T I O N O F 0.5$ L - L Y S I N E TO T H E W H E A T B A S A L D I E T . S U M M A R Y A N D C O N C L U S I O N THE E F F E C T S O F D I E T A R Y P R O T E I N Q U A L I T Y A N D Q U A N T I T Y ON DNA, RNA A N D P R O T E I N A C C U M U L A T I O N I N C H I C K G A S T R O C N E M I U S M U S C L E A N D L I V E R W E R E M E A S U R E D . C H I C K S ( O N E D A Y O F A G E ) W E R E F E D W H E A T A S T H E S O L E S O U R C E O F P R O T E I N , W H E A T - 0 . 5 $ L Y S I N E , W H E A T - S O Y A (15$ P R O T E I N ) A N D 20$ P R O T E I N D I E T S F O R 28 D A Y S . PROTEIN/DNA A N D RNA/DNA R A T I O S I N T H E M U S C L E I N C R E A S E D W I T H I N C R E A S I N G D I E T A R Y P R O T E I N S I N T H E C H I C K F R O M H A T C H TO 28 D A Y S O F A G E . THE R A T I O S I N T H E L I V E R D E C R E A S E D A T 14 D A Y S A N D I N C R E A S E D A T 28 D A Y S O F A G E A S T H E Q U A N T I T Y A N D Q U A L I T Y O F D I E T A R Y P R O T E I N I N C R E A S E D C O N F I R M I N G A S I M I L A R O B S E R V A T I O N W I T H I N C R E A S I N G D I E T A R Y L Y S I N E I N T R I A L 1. R E S U L T S I N D I C A T E T H A T L I V E R G R O W T H I S B Y A N I N C R E A S E I N C E L L N U M B E R ( H Y P E R P L A S I A ) D U R I N G T H E F I R S T 14 D A Y S O F A G E A N D B Y I N C R E A S E S I N C E L L N U M B E R A N D I N C E L L S I Z E ( H Y P E R T R O P H Y ) F R O M 14 TO 28 D A Y S O F A G E W H E R E A S M U S C L E G R O W T H WAS B Y A N I N C R E A S E I N C E L L S I Z E A N D NOT I N T H E N U M B E R O F C E L L S OR F I B R E S F R O M H A T C H TO 28 D A Y S O F A G E T H E I N C R E M E N T S I N M U S C L E A N D L I V E R DNA, RNA A N D P R O T E I N A C C U M U L A T I O N O F C H I C K S F E D LOW P R O T E I N D I E T S F R O M 14 TO 28 D A Y S O F A G E W E R E E S T I M A T E O A N D C O M P A R E D TO T H E C H A N G E I N M U S C L E A N D L I V E R C O N S T I T U E N T S I N C H I C K S F E D T H E 20$ P R O T E I N D I E T . A C C U M U L A T E D M U S C L E A N D L I V E R DNA, RNA A N D P R O T E I N I N C H I C K S F E D T H E LOW P R O T E I N D I E T S WAS L E S S T H A N I N C H I C K S R E C E I V I N G 20$ P R O T E I N D I E T . P R O T E I N A C C U M U L A T I O N WAS A F F E C T E D M ORE S E V E R E L Y I N B O T H O R G A N S T H A N DNA. T H E W H E A T D I E T P E R M I T T E D S I G N I F I C A N T L Y L E S S DNA, RNA A N D P R O T E I N A C C U M U L A T I O N C O M P A R E D W I T H WHEAT-0.5$ L Y S I N E , W H E A T - S O Y A (15$ P R O T E I N ) A N D 20$ P R O T E I N D I E T S T H R O U G H O U T T H E S T U D Y . T H E R E S U L T S I N D I C A T E T H A T , A L T H O U G H C H I C K G R O W T H I S S E V E R E L Y I N H I B I T E D B Y L Y S I N E A N D P R O T E I N D E F I C I E N C Y , M U S C L E A N D L I V E R DNA S Y N T H E S I S C O N T I N U E S A T A F A S T E R R A T E T H A N P R O T E I N S Y N T H E S I S . T H I S WOULD I N D I C A T E T H A T T I S S U E C E L L S I Z E ( E S P E C I A L L Y T H E M U S C L E ) M A Y B E G R E A T L Y R E D U C E D I N C H I C K S U N D E R -N O U R I S H E D F R O M A N E A R L Y A G E B U T NO D E G E N E R A T I O N OR L O S S O F C E L L S OR F I B R E S O C C U R S A N D M A Y E X P L A I N WHY C H I C K S U N D E R S E V E R E P R O T E I N A N D / O R C A L O R I E M A L N U T R I T I O N C A N GROW TO N O R M A L S I Z E W H E N R E H A B I L I T A T E D ON D I E T S A D E Q U A T E I N P R O T E I N A N D C A L O R I E S . TRIAL 4. THE I N F L U E N C E O F D I E T A R Y L Y S I N E ON T H E R A T E O F P R O T E I N S Y N T H E S I S A C C U M U L A T I O N A N D P R O T E I N D E G R A D A T I O N I N T H E T I B I A M I X E D M U S C L E S A N D L I V E R WAS E X A M I N E D I N M O R E D E T A I L . C H A N G E S I N B O D Y W E I G H T A N D L I V E R W E I G H T , A N D T H E A M O U N T S O F P R O T E I N A N D RNA I N T H E M U S C L E S A N D L I V E R S O F C H I C K S F E D T H R E E D I E T A R Y L Y S I N E L E V E L S (0.36, 0.86, 1.24$) A R E S U M M A R I Z E D I N T A B L E S 16 A N D 17 R E S P E C T I V E L Y . B O D Y W E I G H T S I N C R E A S E D P R O G R E S S I V E L Y D U R I N G T H E S T U D Y I N E A C H L Y S I N E T R E A T M E N T . H O W E V E R , R A T E O F I N C R E A S E WAS L E S S I N T H E L Y S I N E D E F I C I E N T G R O U P . |N C O N T R A S T TO T H E P A T T E R N O F B O D Y W E I G H T G A I N , L I V E R W E I G H T D E C L I N E D A F T E R TWO D A Y S I N T H E G R O U P S R E C E I V I N G 0.36 A N D 1.24$ L Y S I N E T R E A T M E N T S W H E R E A S 0.86$ D I E T A R Y L Y S I N E S U P P O R T E D A P R O G R E S S I V E I N C R E A S E I N L I V E R W E I G H T . T H E C O N C E N T R A T I O N O F M U S C L E P R O T E I N ( A V E R A G E F O R E I G H T D A Y S ) E X P R E S S E D A S M G . / G . WET T I S S U E I M P R O V E D (P <0.05) P R O G R E S S I V E L Y A S D I E T A R Y L Y S I N E I N C R E A S E D . M U S C L E RNA C O N T E N T ( M G . / G . ) A L S O I M P R O V E D (P <0.01) B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36 TO 0.86$. NO F U R T H E R I M P R O V E M E N T I N M U S C L E RNA O C C U R R E D B E Y O N D 0.86$ D I E T A R Y L Y S I N E A B S O L U T E L I V E R P R O T E I N A N D RNA C O N T E N T ( M G . / L I V E R ) , A V E R A G E F O R E I G H T D A Y S , I N C R E A S E D (p <0.01) B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36 TO 0.86$ A N D NO F U R T H E R I N C R E A S E O C C U R R E D I N T H E L I V E R C O N S T I T U E N T S B E Y O N D 0.86$ L Y S I N E . L l V E R P R O T E I N A N D RNA C O N T E N T ( A V E R A G E F O R E I G H T D A Y S ) W H E N E X P R E S S E D A S M G . / G . WET T I S S U E R E S P O N D E D T O D I E T A R Y L Y S I N E C O N T E N T I N A M A N N E R C O N T R A R Y TO T H E R E S P O N S E O F A B S O L U T E V A L U E S THE T I S S U E C O N S T I T U E N T S ( M G . / G . W E T T I S S U E ) WERE I N H I G H E R (P < 0.05) A M O U N T S I N L Y S I N E D E F I C I E N C Y T H A N I N C H I C K S F E D 0.86 A N D 1.24$ L Y S I N E . THE R E S P O N S E O F B O T H T H E M U S C L E A N D L I V E R P R O T E I N A N D RNA TABLE 16.--CHANGES I N B O D Y A N D L I V E R W E I G H T S O F C H I C K S F E D G R A D E D 1 4 L E V E L S O F D I E T A R Y L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - C A V E R A G E BODY W E I G H T DAYS O F AGE I N I T I A L F 1 N A L % C H A N G E L I V E R WT. G . G . 0.36 '•>% L Y S I N E 8 59 60 1 .7 1 .5 9 59 62 5.1 2.2 11 61 68 11 .5 2.0 15 55 69 25.4 1 .8 0.8c yfo L Y S I N E 8 76 84 10.5 5.2 9 67 82 22.4 4.5 11 75 110 46.7 5.0 15 75 136 81 .3 6.0 1 .24$ L Y S I N E 8 79 88 1 1 .4 4.6 9 80 96 20.0 6.3 11 75 108 44.0 5.9 15 73 148 102.7 5.8 ^ E A C H V A L U E R E P R E S E N T S A V E R A G E W E I G H T O F T H R E E C H I C K S . TABLE 17.--CHANGES I N P R O T E I N S A N D RNA C O N C E N T R A T I O N I N T H E M I X E D T I B I A M U S C L E S A N D L I V E R OF C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - ^ 4 C LYS 1NE M U S C L E * - L I V E R T R E A T M E N T AGE P R O T E I N RNA** P R O T E I N * RNA* % D A Y S MG . / G . MG./G J MG . / L 1 V E R * * MG./G.' M G . / L 1 V E R * * 8 133.4A 1.9AB 1.9AB 110.oB 167.2A 8.6°° 8.2B° 8.9° 13.2A 9 135.5A 127.3° 288.9A 1 8.2A 0.36 1 1 15 145.2B 147.2B° 140.3 1.8A  1- 7A 137.0E 161.5° 275.6A 290.6A 1 8.0A 18.2A AVE . 1 .8 133.9 255.6A 9.0 16.9 8 9 145.2B 150.0CD 2.8° 1.9AB 114.7B 116.2BC 126.5° 141 AlF 124.7 593.5B 521.9B 637.5B 853.2° 651 .5 7.6B 8.8°° 9 . 2°E 9 . 0°E 8.7B 39.3B 38.9B 46.2B° 54.3B° 44.7B 0.86 11 15 150.0CD 151 .9° 149.3 2.1B 1 9AB 2.2B AVE . 8 149.3C° 2.6C° 124.0C° 567.8B 635.3B 691.2BC ? 8 % °D 39.3B 9 152.4° 2.1B 101 .2A 6.5A 7.9B 9.6-8.2B 40.78 46.3B° 56.2B° 45.7B 1 .24 11 152.2° 1.9AB 117.7BC 15 157.6E 1 52. 9^  2.3B 147.1 J 122.5 862.0° 689.1 AVE . * V A L U E S F O L L O W E D B Y D I F F E R E N T S M A L L L E T T E R S A N D A V E R A G E S F O L L O W E D B Y D I F F E R E N T C A P I T A L L E T T E R S A R E S I G N I F I C A N T L Y D I F F E R E N T (P "^O.OS). * * V A L U E S F O L L O W E D B Y D I F F E R E N T C A P I T A L L E T T E R S A R E S I G N I F I C A N T L Y D I F F E R E N T (P •< 0 . 0 1 ) . ''DATA R E P R E S E N T M G . / G . WET T I S S U E . C O N S T I T U E N T S TO D I E T A R Y L Y S I N E O N C E A G A I N D E M O N S T R A T E S T H A T G R O W T H I N M U S C L E S I Z E A N D M U L T I P L I C A T I O N OF L I V E R C E L L S O C C U R I N T H E C H I C K A T 14 D A Y S O F A G E A S O B S E R V E D I N T R I A L S 1 A N D 3. M U S C L E P R O T E I N I N C R E A S E D ( T A B L E 17) A S C H I C K G R O W T H P R O G R E S S E D ( T A B L E 16) I R R E S P E C T I V E O F D I E T A R Y L Y S I N E L E V E L . S I M I L A R A M O U N T S O F L I V E R P R O T E I N A N D RNA A T 0.86 A N D 1.24$ D I E T A R Y L Y S I N E A G R E E W I T H R E S U L T S I N T R I A L 1 T H A T P R O T E I N A N D RNA C O N T E N T O F T H E C H I C K L I V E R I N C R E A S E D S I G N I F I C A N T L Y F R O M 0.36 TO 0.86$ D I E T A R Y L Y S I N E W I T H NO D I F F E R E N C E B E T W E E N C H I C K S R E C E I V I N G 0.86 A N D 1.11$ L Y S I N E . H I G H E R L I V E R P R O i T E I N ( M G . / G . WET T I S S U E ) I N T H E L Y S I N E D E F I C I E N T C H I C K T H A N I N C H I C K S F E D 0.86 OR 1.24$ D I E T A R Y L Y S I N E S U P P O R T S T H E V I E W ( T R I A L S 1, 3) T H A T P R O T E I N S I N T H E L I V E R O F C H I C K S F E D A D I E T S E V E R E L Y D E F I C I E N T I N L Y S I N E R E P R E S E N T A C A R R Y O V E R F R O M T H E E G G T H A T I S S L O W L Y M E T A B O L I Z E D D U E TO S L O W G R O W T H . GROWTH R A T E , M U S C L E P R O T E I N A N D RNA C O N T E N T I N T H E C H I C K F E D 1^24$ L Y S I N E W E R E E X P E C T E D TO B E L E S S T H A N I N C H I C K S C O N S U M I N G 0.86$ L Y S I N E S I N C E 1.11$ L Y S I N E D E P R E S S E D G R O W T H A N D M U S C L E P R O T E I N A N D RNA I N T R I A L 1. HOWEVER, C H I C K S U S E D I N T H I S T R I A L W E R E M O R E T O L E R A N T TO H I G H L Y S I N E (1.24$) I N T H E D I E T . F E E D I N T A K E S O F C H I C K S R E C E I V I N G 0.36, 0.86 A N D 1.24$ L Y S I N E F O R 15 D A Y S W E R E O F T H E O R D E R 73, 132 A N D 152 G . / C H I C K . IN C O N T R A S T , F E E D I N T A K E S O F C H I C K S R E C E I V I N G 0.36, 0.86 A N D 1.11$ L Y S I N E F O R 14 D A Y S I N T R I A L 1 W E R E 92, 211 A N D 191 G . / C H I C K R E S P E C T I V E L Y . T H E D E C R E P A N C Y I N F E E D I N T A K E WAS P R O B A B L Y D U E TO A D I F F E R E N C E I N T H E N U M B E R O F C H I C K S P E R T R E A T M E N T U S E D I N E A C H T R I A L A N D C H A N G E S I N E N V I R O N M E N T A L T E M P E R A T U R E . T W E L V E C H I C K S P E R T R E A T M E N T W E R E U S E D I N T H E P R E S E N T T R I A L W H E R E A S 36 C H I C K S P E R T R E A T M E N T W E R E U S E D I N T R I A L 1. T H E P R E S E N T T R I A L WAS R U N I N S U M M E R ( A U G U S T , 1972) W H E R E A S T R I A L 1 WAS R U N I N W I N T E R ( D E C E M B E R , 1971/ J A N U A R Y , 1972). HOWEVER, I T I S NOT KNOWN I F T H E A D M I N I S T R A T I O N O F L Y S I N E - U - C I N T O T H E C H I C K S D I D I N F L U E N C E F E E D I N T A K E . F I G U R E S 19 A N D 20 I L L U S T R A T E T H E A C C U M U L A T I O N O F P R O T E I N A N D RNA I N T H E M U S C L E A N D L I V E R S R E S P E C T I V E L Y O F C H I C K S F E D T H E T H R E E 1 4 L E V E L S O F L Y S I N E (0.36, 0.86, 1.24$) D U R I N G L Y S I N E - U - C A D M I N I S T R A -T I O N . M U S C L E P R O T E I N ( M G . / G . ) I N C R E A S E D G R A D U A L L Y D U E TO L Y S I N E T R E A T M E N T S . M U S C L E RNA ( M G . / G . ) WAS I R R E G U L A R R E S U L T I N G I N A N I R R E G U L A R P R O T E I N / R N A R A T I O . L I V E R P R O T E I N ( F I G . 20) A C C U M U L A T E D I N A L I N E A R M A N N E R A T T H E R A T E O F 37.1 A N D 42.0 M G . / D A Y OR 4.3 A N D 4.9$ P E R D A Y I N C H I C K S R E C E I V I N G 0.86 A N D 1.24$ L Y S I N E R E S P E C T I V E L Y . P R O T E I N I N T H E L I V E R O F C H I C K S F E D L Y S I N E D E F I C I E N T D I E T (0.36$) I N C R E A S E D I N A C U R V E -L I N E A R M A N N E R A T T H E R A T E O F A P P R O X I M A T E L Y 17.6 MG. OR 6$ P E R D A Y . T H E D A I L Y R A T E O F L I V E R P R O T E I N A C C U M U L A T I O N WAS E S T I M A T E D B Y D I V I D I N G T H E D I F F E R E N C E B E T W E E N L I V E R P R O T E I N C O M P O S I T I O N ON T H E E I G H T H A N D T H A T ON T H E F I R S T D A Y A F T E R R A D I O I S O T O P E I N J E C T I O N B Y 7 ( P E R I O D O F S T U D Y ) . THE R E S U L T I N G V A L U E E X P R E S S E D A S A P E R C E N T A G E O F T H E V A L U E ON T H E E I G H T H D A Y F O L L O W I N G R A D I O I S O T O P E A D M I N I S T R A T I O N G A V E T H E P E R C E N T A G E R A T E O F A C C U M U L A T I O N . L L V E R RNA A C C U M U L A T E D I N A S I M I L A R M A N N E R TO T H E P R O T E I N . P R O T E I N / R N A R A T I O WAS C O N S T A N T I R R E S P E C T I V E O F L Y S I N E L E V E L D E M O N S T R A T I N G O N C E A G A I N T H E C O R R E L A T I O N O F RNA TO L I V E R P R O T E I N S Y N T H E S I S . T H E I N C O R P O R A T I O N O F L Y S I N E - U - C A C T I V I T Y I N T O T H E L I V E R A N D M U S C L E O F C H I C K S F E D T H E T H R E E L E V E L S (0.36, 0.86, 1.24$) O F L Y S I N E 14 I S S U M M A R I Z E D I N T A B L E 18. A C T I V I T Y O F L Y S I N E - U - C E X P R E S S E D A S X —' 1 1 1 1 1 I I * I 1 2 3 4 5 6 7 8 9 10 DAY F O L L O W I N G A D M I N I S T R A T I O N O F L Y S I N E - U - 4C M U S C L E P R O T E I N , RNA A N D P R O T E I N / R N A O F C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E A N D I N J E C T E D W I T H L Y S I N E -u - 1 4 c . 117 F I G . 20. L I V E R P R O T E I N , RNA A N D P R O T E I N / R N A R A T I O I N C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - 1 4 C . TABLE 18.--EFFECT O F D I E T A R Y L Y S I N E ON T I S S U E P R O T E I N D E G R A D A T I O N DAYS L I V E R * M U S C L E *  A F T E R R A D I P A C T I V I T Y S P E C I F I C A C T I V I T Y R A D I P A C T I V I T Y S P E C I F I C A C T I V I T Y 14c I N J E C T I C N D P M / G . T I S S U E D P M / M G . P R P T E I N D P M / G . T I S S U E D P M / M G . P R D T E I N 0.36$ L Y S I N E 1 23,312.1A 212.2A 15,416.6A 115.6A 2 20,045.6B 157.78 13,880.2B 102.4B 4 17,534.5° 128.2° 12,429.3° 85.7° 8 16,833.4° 104.3° 13,099.5° 90.5° T O T A L * * 77,725.6A 602.4 54,815.6 394.2 0.86$ L Y S I N E 1 21,586.2A 189.1A 14,729.0A 101.6A 2 19,088.7B 164.4B 16,887.3S 112.6s 4 11,044.9° 87.5° 11,586.1° 77.3° 8 8,376.3° 59.3° 10,293.5° 70.9° T O T A L * * 60,096.1 500.3 53,962.8 362.4 1.24$ L Y S I N E 1 19,128.2A 154.4A 14,751.5A 98.9A 2 13,172.88 130.0B 12,125.0S 79.6s 4 11,338.9° 94.3° 11,795.8S 77.5B 8 6,212.2° 42.2° 10,293.5° 65.3° T O T A L * * 49,852.1° 420.9° 48,965.8° 321.3° * V A L U E S F O L L O W E D B Y A D I F F E R E N T S M A L L L E T T E R A R E S I G N I F I C A N T L Y D I F F E R E N T (P <0.01). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T C A P I T A L L E T T E R A R E S I G N I F I C A N T L Y D I F F E R E N T (P *=0.01). CO R A D I O A C T I V I T Y ( D P M / G . T I S S U E ) OR S P E C I F I C A C T I V I T Y ( D P M / M G . T I S S U E P R O T E I N ) D E C R E A S E D P R O G R E S S I V E L Y A T A S I G N I F I C A N T (P *=0.01) R A T E B O T H I N T H E L I V E R A N D I N T H E M U S C L E F R O M T H E D A Y O F A D M I N I S T R A T I O N TO T H E E I G H T H D A Y O F A D M I N I S T R A T I O N . T O T A L R A D I O A C T I V E L Y S I N E I N C O R P O R A T E D I N T O T H E L I V E R A N D M U S C L E D U R I N G T H E S T U D Y D E C R E A S E D S I G N I F I C A N T L Y (:P <0.01) W I T H H I G H E R L E V E L S O F D I E T A R Y L Y S I N E . P R E V I O U S S T U D I E S ( S I D R A N S K Y A N D F A R B E R , 1958; N I M N I A N D B E V E T T A , 14 1961) H A V E D E M O N S T R A T E D T H A T R A T S F E D A C L A B E L L E D T H R E O N I N E - OR T R Y P T O P H A N - D E F I C I E N T D I E T HAD H I G H E R S P E C I F I C A C T I V I T Y O F L I V E R P R O T E I N S T H A N T H E G R O U P C O N S U M I N G T H E S U P P L E M E N T E D D I E T . |T H A S A L S O B E E N R E P O R T E D ( R O D R I G U E Z , 1969) T H A T L I V E R C E L L S A P A N D M I C R O S O M E S I S O L A T E D F R O M R A T S F E D W I T H W H E A T G L U T E N I N C O R P O R A T E D 158$ M O R E 1 4 „ L E U C I N E - C T H A N P R E P A R A T I O N S F R O M C A S E I N - F E D A N I M A L S . T H E M I C R O S O M E S W E R E B E L I E V E D TO B E P R I M A R I L Y R E S P O N S I B L E F O R T H E I N C R E A S E . R A D I O A C T I V I T I E S A N D S P E C I F I C A C T I V I T I E S W E R E H I G H E R ON T H E F I R S T 1 4 „ D A Y O F L Y S I N E - U - C A D M I N I S T R A T I O N I N C H I C K S T H A T W E R E F E D T H E L Y S I N E D E F I C I E N T D I E T (0.36$) T H A N I N T H O S E F E D 0.86 A N D 1.24$ L Y S I N E D I E T S I N D I C A T I N G A N E E D F O R L Y S I N E F O R G R O W T H O F C H I C K S F E D 0.36$ L Y S I N E . THE R E S U L T S A G R E E W I T H P R E V I O U S O B S E R V A T I O N S W I T H R A T S F E D H I G H P R O T E I N D I E T S T H A T HAD L O W E R S P E C I F I C A C T I V I T I E S I N T H E L I V E R , P L A S M A , K I D N E Y A N D I N T E S T I N A L M U S C O S A P R O T E I N S T H A N R A T S R E C E I V I N G LOW P R O T E I N D I E T S ON T H E F I R S T D A Y O F R A D I O I S O T O P E A D M I N I S T R A T I O N ( S O L O M O N A N D T A R V E R , 1952; A L L I S O N A N D W A N N E M A C H E R , 1958; WATERLOW, 1959; W A N N E M A C H E R , 1961 A N D M U R A M A T S U _ET _ A L . 1963). 1 4 B I O L O G I C A L H A L F - L I F E ( T ^ ) O F L Y S I N E - C I N T H E L I V E R P R O T E I N S . WAS'E E S T I M A T E D I N T H E P R E S E N T S T U D Y B Y P L O T T I N G T H E L O G A R I T H M O F S P E C I F I C A C T I V I T Y OF P R O T E I N ( D P M / G . P R O T E I N ) A S A F U N C T I O N O F T I M E ( F I G U R E 21). THE S L O P E O F T O T A L R A D I O A C T I V I T Y G A V E T H E D E G R A D A T I O N R A T E A N D T H E F O R M U L A TG/II.N 2 W H I C H I S E Q U I V A L E N T TO TG/2.3 L O G 2 ( O R T^/O.693) G A V E T H E T U R N O V E R T I M E C O N S T A N T F O R D E G R A D A T I O N . THE D A I L Y F R A C T I O N A L C A T A B O L I C R A T E A S A P E R C E N T A G E O F T H E T O T A L P R O T E I N WAS D E R I V E D F R O M 100 l l N 2/T^. THE T U R N O V E R R A T E C O N S T A N T O F S Y N T H E S I S WAS C A L C U L A T E D A S D E S C R I B E D F O R RNA B Y ENWONWU _ET _ A L . (1971) F R O M T H E R A T E C O N S T A N T OF D E G R A D A T I O N A N D A M O U N T O F T O T A L P R O T E I N P R E S E N T 14 D U R I N G T H E E X P E R I M E N T . T H E F I R S T D A Y F O L L O W I N G L Y S I N E - U - C I N J E C T I O N WAS T A K E N A S T I M E To A N D T H E S E V E N T H D A Y A S T1 . AT TO T O T A L L I V E R P R O T E I N = Ro E Q U A T I O N 1 AT T1 A M O U N T O F P R O T E I N D E G R A D E D B E T W E E N TO A N D T1 C A L L E D D1 = RX W H E R E X = D A I L Y F R A C T I O N A L C A T A B O L I C R A T E A N D D E R I V E D F R O M R A D I O A C T I V I T Y P L O T . AT T1 T O T A L L I V E R P R O T E I N = R1 R1 = (1 + Y) RO E Q U A T I O N 2 W H E R E Y = D A I L Y F R A C T I O N A L C H A N G E I N T O T A L P R O T E I N . S I N C E S Y N T H E S I S O F NEW P R O T E I N M O L E C U L E D O E S N O T C E A S E B E T W E E N TO A N D T1, R1 - S1 = Ro - D1 E Q U A T I O N 3 W H E R E S1 = A M O U N T O F N E W L Y S Y N T H E S I Z E D P R O T E I N B E T W E E N TO A N D T1. B Y S U B S T I T U T I N G E Q U A T I O N S 1 A N D 2 I N E Q U A T I ON 3, (1 + Y) RO - S1 = Ro - xRo THUS SI = YRO + xRo. L I V E R P R O T E I N S P E C I F I C A C T I V I T Y C U R V E S ( F I G . 21) D E C R E A S E D E X P O N E N T I A L L Y R E A C H I N G H A L F T H E I N I T I A L V A L U E S I N 7.2, 4.9 A N D 5.2 D A Y S R E S P E C T I V E L Y F O R C H I C K S F E D 0.36, 0.86 A N D 1.24$ D I E T A R Y L Y S I N E . THE H A L F - L I F E V A L U E S C O R R E S P O N D TO 10.4, 7.1 A N D 7.5 D A Y S T U R N O V E R T I M E A N D N E T D A I L Y D E G R A D A T I O N O F 9.6, 14.0 A N D 13.3$. T O T A L P R O T E I N S Y N T H E S I Z E D I N T H E L I V E R D U R I N G T H E S T U D Y (7 D A Y S ) WAS E S T I M A T E D A T 5.4 4.41 l 1 1 1 I i 1 2 3 4 5 6 7 8 9 10 DAY F O L L O W I N G A D M I N I S T R A T I O N O F L YS I NE-U-''4C 1 4 F I G . 21. RATE O F L O S S I N C - P R O T E I N F R O M L I V E R S O F C H I C K S R E C E I V I N G G R A D E D L E V E L S O F D I E T A R Y L Y S I N E . 139.8, 344.2 A N D 370.7 MG. ( I . E . 20, 49.2 A N D 52.9 M G . / D A Y ) F O R 0.36, 0.86 A N D 1.24$ L Y S I N E G R O U P S . T H E S L O W E R T U R N O V E R T I M E A N D L O W E R S Y N T H E T I C R A T E O F L I V E R P R O T E I N S I N L Y S I N E - D E F I C I E N T C H I C K S T H A N I N C H I C K S R E C E I V I N G A D E Q U A T E L Y S I N E C O N F I R M T H E V I E W ( T R I A L S 1 A N D 3) T H A T L I V E R P R O T E I N S H I G H E R T H A N N O R M A L I N C H I C K S F E D 0.36$ L Y S I N E P R O B A B L Y R E P R E S E N T R E S I D U A L P R O T E I N F R O M T H E E G G . E S T I M A T I O N O F L I V E R P R O T E I N S S Y N T H E S I Z E D D U R I N G T H E S T U D Y B Y U S I N G T H E F O R M U L A R E P O R T E D B Y ENWONWU E T A L . (1971) G A V E H I G H E R V A L U E S T H A N B Y T A K I N G T H E D I F F E R E N C E B E T W E E N L I V E R P R O T E I N C O M P O S I T I O N ON T H E E I G H T H A N D T H A T ON T H E F I R S T D A Y A F T E R R A D I O I S O T O P E I N J E C T I O N D E S C R I B E D E A R L I E R . T H E F O R M U L A O F ENWONWU _ET _ A L . (1971) G A V E A V A L I D E S T I M A T E S I N C E P R O T E I N S D E G R A D E D A N D S Y N T H E S I Z E D D A I L Y T H R O U G H O U T T H E S T U D Y A R E C O N S I D E R E D I N T H E F O R M U L A . E S T I M A T E D H A L F - L I F E V A L U E S O F L I V E R P R O T E I N R E P O R T E D I N T H E L I T E R A T U R E V A R I E D F R O M 2.8 TO 7 D A Y S ( S H E M I N A N D R L T T E N B E R G , 1944; FR 1 E D B E R G _E_T _ A L . 1948, T A R V E R A N D MO R S E , 1 948; N E U B E R G E R _ET _ A L . 1951; T H O M P S O N A N D B A L L O U , 1956; M U R A M A T S U _ET _ A L . 1963; H I R S C H A N D H I A T T , 1966). H A L F - L I F E V A L U E S O F L I V E R P R O T E I N I N T H E P R E S E N T S T U D Y F A L L W I T H I N T H E R A N G E R E P O R T E D . R E S U L T S D E M O N S T R A T E T H A T T H E R A T E O F P R O T E I N S Y N T H E S I S A N D D E G R A D A T I O N I N T H E L I V E R O F L Y S I N E D E F I C I E N T C H I C K S WAS C O N S I D E R A B L Y L E S S T H A N I N T H E L I V E R O F C H I C K S C O N S U M I N G A D E Q U A T E L Y S I N E I N D I C A T I N G A G A I N A R E Q U I R E M E N T O F L Y S I N E F O R P R O T E I N S Y N T H E S I S . THE L O G A R I T H M O F S P E C I F I C A C T I V I T Y O F M U S C L E P R O T E I N ( D P M / G . P R O T E I N ) WAS P L O T T E D A S A F U N C T I O N O F T I M E . THE R E S U L T I N G C U R V E S F R O M T H E P L O T F O R C H I C K S F E D D I E T A R Y L E V E L S O F 0.36, 0.86 A N D 1 L Y S I N E A R E I L L U S T R A T E D I N F l G . 22. M U S C L E P R O T E I N S P E C I F I C A C T I V I T Y I R R E S P E C T I V E O F T H E L E V E L O F D I E T A R Y L Y S I N E D E C R E A S E D I N A C U R V E L I N E A R 14, M A N N E R F R O M T H E F I R S T TO T H E E I G H T H D A Y O F L Y S I N E - U - C A D M I N I S T R A T I O N . THE C U R V E L I N E A R P A T T E R N O F P R O T E I N S P E C I F I C A C T I V I T Y A S A F U N C T I O N O F T I M E M A D E T H E E S T I M A T I O N O F H A L F - L I F E V A L U E S O F M U S C L E P R O T E I N S D I F F I C U L T . H OWEVER, M U S C L E P R O T E I N S I N C H I C K S F E D 0.86 A N D 1.24$ D I E T A R Y L Y S I N E D I D T U R N O V E R MORE R A P I D L Y T H A N I N C H I C K S R E C E I V I N G T H E L Y S I N E D E F I C I E N T D I E T ( F L G . 22 A N D 23). A V A I L A B L E R E P O R T S ON T H E R A T E O F L O S S O F M U S C L E T I S S U E P R O T E I N S P E C I F I C A C T I V I T Y I N T H E L I T E R A T U R E A R E I N C O N S I S T E N T . T H E H A L F - L I F E O F M U S C L E P R O T E I N S I N T H E R A T C O U L D NOT B E C A L C U L A T E D B Y M U R A M A T S U E_T _ A L . (1963) B E C A U S E T H E C H A N G E I N T H E S P E C I F I C A C T I V I T Y O F T H E M U S C L E P R O T E I N S D U R I N G T H E S E V E N D A Y S O F S T U D Y WAS N O T S I G N I F I C A N T , N E U B E R G E R _ET _ A L . (1951), ON T H E O T H E R H A N D , O B S E R V E D T H A T M I X E D P R O T E I N S O F M U S C L E I N Y O U N G R A T S HAD A H A L F - L I F E O F 24 TO 30 D A Y S . THE D A T A O F T H O M P S O N A N D B A L L O U (1956) A L S O I N D I C A T E D T H A T 60$ O F M U S C L E P R O T E I N S HAD A H A L F - L I F E O F 16 D A Y S W H I L E T H E R E S T HAD A B O U T 100 D A Y S . S T U D I E S O F I S O T O P E I N C O R P O R A T I O N I N T O I S O L A T E D M U S C L E P R O T E I N S L E D V E L I C K (1956) TO C O N C L U D E T H A T I N D I V I D U A L M U S C L E P R O T E I N S T U R N E D O V E R A T M A R K E D L Y D I F F E R E N T R A T E S . H E C O M P U T E D T H A T F O R T H E R A T T H E H A L F - L I F E O F S K E L E T A L M U S C L E G L Y C E R A L D E H Y D E - 3 - P H O S P H A T E D E H Y D R O G E N A S E WAS 100 D A Y S , H E A V Y M E R O M Y O S I N G A V E 80 D A Y S , A C T I N WAS 67 D A Y S , A L D O L A S E A N D P H O S P H O R Y L A S E WERE B O T H 50 D A Y S , T R O P O M Y O S I N WAS 27 D A Y S A N D L I G H T M E R O M Y O S I N WAS 20 D A Y S . ON T H E O T H E R H A N D , M C M A N U S A N D M U E L L E R (1966) O B S E R V E D T H A T T H E T U R N O V E R T I M E S O F L I G H T A N D H E A V Y M E R O M Y O S I N F O R R A B B I T S K E L E T A L M U S C L E W E R E B O T H 29 D A Y S . 5.2 124 o o_ o co CL Q C5 O 5.0 0.36$ L Y S I N E 4.8 0.86$ L Y S I N E ^•Al .24$ L Y S I N E 4.61 -1_ -U _1_ F I G . 22. 2 3 4 5 6 7 8 9 10 D A Y F O L L O W I N G A D M I N I S T R A T I O N O F L Y S I N E 14, u - 1 4 c RATE O F L O S S O F ' C - P R O T E I N F R O M T H E T I B I A M U S C L E O F C H I C K S F E D G R A D E D L E V E L S OF D I E T A R Y L Y S I N E . 125 F I G . 23. C H A N G E S I N T H E P E R C E N T A G E D E C R E A S E O F S P E C I F I C A C T I V I T I E S O F T H E L I V E R A N D M I X E D T I B I A M U S C L E T I S S U E P R O T E I N S O F C H I C K S F E D G R A D E D L E V E L S O F D I E T A R Y L Y S I N E . 126 F R I T Z _ E T _ A L . (1969) O B T A I N E D A H A L F - L I F E O F 31 A N D 21 D A Y S F O R L A C T A T E D E H Y D R O G E N A S E - 5 A N D T O T A L S O L U B L E P R O T E I N R E S P E C T I V E L Y I N S K E L E T A L M U S C L E B Y F O L L O W I N G A C O N T I N U O U S D I E T A R Y A D M I N I S T R A T I O N O F A 1 4 C - L A B E L L E D Y E A S T P R O T E I N H Y D R O L Y S A T E TO A D U L T R A T S . M l L L W A R D (1970) I N J E C T E D C - L A B E L L E D C A R B O N A T E I N T O R A T S A N D E S T I M A T E D T H E F R A C T I O N A L R A T E O F C A T A B O L I S M F O R T H E S A R C O P L A S M I C A N D M Y O F I B R I L L A R F R A C T I O N O F S K E L E T A L M U S C L E A S D E T E R M I N E D F R O M T H E D E C A Y R A T E O F A S P A R T A T E A N D G L U T A M A T E I S O L A T E D F R O M T H E P R O T E I N F R A C T I O N S . HE O B S E R V E D A H A L F - L I F E O F 3.6 A N D 15.6 D A Y S F O R T H E S A R C O P L A S M I C A N D M Y O F I B R I L L A R P R O T E I N S R E S P E C T I V E L Y . Y O U N G JET_ _AL_. (1971 ) S H O W E D T H A T T H E H A L F - L I F E F O R T H E M I X E D M U S C L E P R O T E I N S WAS 13.1 D A Y S A S C O M P A R E D W I T H A V A L U E O F 10.2 D A Y S F O R D E P L E T E D R A T S W I T H I N T R A V E N O U S I N J E C T I O N O F A S P A R T I C AC I D -14 4- C. THE P R O B L E M O F M E A S U R I N G P R O T E I N D E G R A D A T I O N R A T E S I N T H E S K E L E T A L M U S C L E A N D I N T H E O T H E R T I S S U E S I S C O M P L I C A T E D B Y R E U T I L I Z A T I ON O F A M I N O A C I D S L I B E R A T E D F R O M D E G R A D A T I O N O F P R O T E I N W I T H I N T H E C E L L . THE R E U T I L I Z A T I O N O F E N D O G E N O U S A M I N O A C I D I N T H E L I V E R I S F R O M 50 TO 90$ (GAN A N D J E F F A Y , 1967; WATERLOW A N D S T E P H E N , 1968) O F T H E T O T A L A M O U N T I N C O R P O R A T E D I N T O P R O T E I N A N D T H I S V A R I E S A C C O R D I N G T O N U T R I T I O N C I R C U M S T A N C E S . A M I N O A C I D R E U T I L I Z A T I O N I N S K E L E T A L M U S C L E I S A B O U T 10 TO 30$ (GAN A N D J E F F A Y , il'967; WATERLOW A N D S T E P H E N , 1968). As A R E S U L T , S I G N I F I C A N T E R R O R S I N E S T I M A T E S O F D E G R A D A T I O N R A T E S F O R T I S S U E P R O T E I N S M A Y A R I S E , S I N C E T H E L O S S O F T H E R A D I O A C T I V I T Y I N P R O T E I N I S L E S S R A P I D T H A N T H E T R U E R A T E O F T U R N O V E R . T H E R E S U L T S O F T H E P R E S E N T S T U D Y , H O W E V E R , D E M O N S T R A T E T H A T M I X E D M U S C L E P R O T E I N S O F C H I C K S D I D T U R N O V E R M O R E R A P I D L Y A N D A G R E A T E R A M O U N T WAS D E G R A D E D D A I L Y A S T H E 127 L E V E L O F D I E T A R Y L Y S I N E I N C R E A S E D (FIG. 22 A N D 23). A R A P I D P R O T E I N T U R N O V E R W I T H A H I G H M U S C L E P R O T E I N C O N C E N T R A T I O N ( T A B L E 17) I N C H I C K S F E D T H E 0.86 OR 1.24$ L Y S I N E D I E T I N D I C A T E A N A C T I V E M U S C L E P R O T E I N S Y N T H E S I S A N D D E G R A D A T I O N . F I G U R E 23 WAS O B T A I N E D B Y P L O T T I N G T H E P E R C E N T A G E D E C R E A S E O F T H E S P E C I F I C A C T I V I T Y I N P R O T E I N S O F T H E L I V E R A N D T H E M U S C L E D U R I N G T H E P E R I O D F R O M T H E F I R S T TO T H E E I G H T H D A Y A G A I N S T T H E L E V E L O F D I E T A R Y L Y S I N E . T H I S F I G U R E I N D I C A T E S T H A T T H E R A T E O F L O S S O F T H E L A B E L F R O M T H E P R O T E I N S I N B O T H T H E L I V E R A N D M U S C L E T I S S U E S I N C R E A S E D W I T H I N C R E A S I N G L E V E L S O F L Y S I N E I N T H E D I E T . T H E L O S S O F T H E L A B E L F R O M T H E T I S S U E P R O T E I N S WAS A T A G R E A T E R R A T E W H E N T H E L E V E L O F D I E T A R Y L Y S I N E WAS I N C R E A S E D F R O M 0.36 TO 0.86$ T H A N W H E N I T WAS I N C R E A S E D F R O M 0.86 TO 1.24$. T H E P A T T E R N O F T H E S E C H A N G E S R E S E M B L E D T H O S E O F T H E B O D Y W E I G H T A S S H O W N I N F L G . 24. T H E S E R E S U L T S A R E I N A G R E E M E N T W I T H P R E V I O U S R E P O R T S T H A T T H E T U R N O V E R R A T E S O F P L A S M A P R O T E I N I N D O G S ( Y U L L E E J _ A L . 1959) A N D O F L I V E R A N D I N T E S T I N A L M U C O S A P R O T E I N S I N R A T S (M U R AM A T S U _ET _AL . 1963) R E C E I V I N G A D E Q U A T E D I E T A R Y P R O T E I N WERE M O R E R A P I D T H A N I N T H O S E R E C E I V I N G A D E F I C I E N T P R O T E I N 35, D I E T . THE R A T E S O F L O S S O F S F R O M T H E P R O T E I N S I N S E V E R A L T I S S U E S S U C H A S I N T E S T I N A L M U C O S A , P L A S M A , K I D N E Y , S P L E E N A N D L I V E R W E R E A L S O O B S E R V E D B Y M U R A M A T S U _ET _ A L . (1963) TO O C C U R I N A S I M I L A R M A N N E R A S T H E C H A N G E S I N B O D Y W E I G H T W I T H I N C R E A S I N G L E V E L S O F C A S E I N I N T H E D I E T . R E S U L T S I L L U S T R A T E D I N F I G . 23 A N D 24 S U G G E S T A C O R R E L A T I O N B E T W E E N T H E P A T T E R N O F T H E G R O W T H R E S P O N S E A N D T H E R E S P O N S E C U R V E O F T H E T U R N O V E R R A T E O F B O T H T H E L I V E R A N D M U S C L E T I S S U E P R O T E I N S . T H E 128 F I G . 24. BODY W E I G H T G A I N P E R C H I C K P E R D A Y O F F E E D I N G G R A D E D L E V E L S O F D I E T A R Y L Y S I N E . I N C R E A S E D T U R N O V E R R A T E O F T I S S U E P R O T E I N W I T H I N C R E A S I N G L E V E L O F D I E T A R Y L Y S I N E I N D I C A T E D A C T I V E P R O T E I N S Y N T H E S I S A N D D E G R A D A T I O N T H A T R E S U L T E D I N T I S S U E P R O T E I N A C C U M U L A T I O N A N D H E N C E T H E I N C R E A S E I N B O D Y WE I G H T . R E S U L T S P R E S E N T E D I N T A B L E 19 D E A L W I T H T H E R A D I O A C T I V I T Y I N C O R P O R A T I O N O F L Y S I N E - U - C I N T O L I V E R A N D M U S C L E P R O T E I N S I N C H I C K S F E D D I E T S C O N T A I N I N G 0.36, 0.86 A N D 1.24$ L Y S I N E F O R 14 D A Y S . THE V A L U E S I N T H E T A B L E D E N O T E T H E A C T I V I T Y R E C O V E R E D I N T I S S U E P R O T E I N S E X P R E S S E D A S A P E R C E N T A G E O F T H E I N I T I A L D O S E , A N D A C T I V I T Y E X P R E S S E D R E L A T I V E TO T I S S U E RNA C O N C E N T R A T I O N . P E R C E N T A G E R E C O V E R Y O F R A D I O A C T I V I T Y I N T H E P R O T E I N F R A C T I O N I N T H E L I V E R A N D M U S C L E D E C R E A S E D S I G N I F I C A N T L Y (P ^ O . O L ) W I T H I N C R E A S I N G D I E T A R Y L Y S I N E . S P E C I F I C A C T T V I T Y , D P M / M G . RNA, A L S O D E C R E A S E D S I G N I F I C A N T L Y I N T H E L I V E R (:P "=0.01) A N D T H E M U S C L E (P < 0.05) W I T H I N C R E A S I N G L Y S I N E . L O W E R P E R C E N T A G E R E C O V E R Y O F A C T I V I T Y I N T I S S U E P R O T E I N S A N D A C T I V I T Y / T I S S U E RNA W I T H 0.86 A N D 1.24$ L Y S I N E T H A N 0.36$ L Y S I N E D O E S N O T R E F L E C T A L O W E R S Y N T H E S I S S I N C E T H E P O O L S I Z E O F A M I N O A C I D I N F L U E N C E T H E I N C O R P O R A T I O N O F R A D I O A C T I V E A M I N O A C I D S I N T O P R O T E I N ( D I L U T I O N E F F E C T ) . |N F A C T , L O W E R P R O T E I N S Y N T H E S I S WAS F O U N D I N T H E L I V E R O F L Y S I N E D E F I C I E N T (0.36$) C H I C K S T H A N I N T H O S E F E D 0.86 A N D 1.24$ L Y S I N E . THE D E C L I N I N G P A T T E R N O F R A D I O A C T I V I T Y R E C O V E R Y A N D S P E C I F I C A C T I V I T Y , D P M / M G . RNA W I T H T H E L E V E L O F D I E T A R Y L Y S I N E , O N C E A G A I N , I N D I C A T E A N I N C R E A S E D R A T E O F T I S S U E P R O T E I N T U R N O V E R A S T H E L E V E L O F D I E T A R Y L Y S I N E I N C R E A S E D . M O R E O V E R , L O W E R P E R C E N T A G E R E C O V E R Y O F A C T I V I T Y I N T I S S U E P R O T E I N S W I T H 0.86 A N D 1.24$ D I E T A R Y L Y S I N E T H A N W I T H 0.36$ L Y S I N E A G R E E S W I T H 1 4 TABLE 1 9 . - - E F F E C T O F D I E T A R Y L Y S I N E L E V E L ON I N C O R P O R A T I O N O F L Y S I N E - U - C I N T O T H E T I S S U E S O F C H I C K S L i V E R * * M U S C L E * * D P M / G . P R O T E I N 1 „2 D P M / L I V E R P R O T E I N D P M / G . P R O T E I N 7~T2 L Y S I N E T R E A T M E N T {%) - 0 ^ - -• - — » X U R L E V E L 2.22 x 10° T O T A L RNA 2.22 x 10° D P M / M G . RNA S U P P . 0.50 0.88 0.36 0.86 1 .24 6.73 288.82 x 10 3' 5.58 4.69 B 215.19 x 10" 4.39 4.03 B 186.14 x 10 3^ 3.57' 7,475.0 6,217.5 E 5,434.1 * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P *= 0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P "=0.01). 131 T H E O B S E R V A T I O N O F Y A M A S H I T A A N D A S H I D A (1969) T H A T R A T S F E D A L Y S I N E F R E E D I E T I N C O R P O R A T E D MORE I N J E C T E D L Y S I N E - U - C I N T O L I V E R • P R O T E I N S T H A N R A T S F E D A D E Q U A T E L Y S I N E . S U M M A R Y A N D C O N C L U S I O N THE E F F E C T S O F D I E T A R Y L Y S I N E L E V E L ON T H E R A T E O F L I V E R A N D M U S C L E P R O T E I N S Y N T H E S I S A N D D E G R A D A T I O N I N T H E C H I C K W E R E S T U D I E D . C H I C K S W E R E F E D T H R E E L E V E L S (0.36, 0.86 A N D 1.24$) O F A D I E T A R Y 14. L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - C I N T R A P E R I T O N E A L L Y . THE T U R N O V E R O F L I V E R A N D M U S C L E P R O T E I N S WAS E X A M I N E D B Y O B S E R V I N G T H E R A T E O F L O S S O F R A D I O A C T I V I T Y O V E R S E V E R A L D A Y S . H A L F - L I F E V A L U E S O F 7.2, 4.9 A N D 5.2 D A Y S C O R R E S P O N D I N G T O 10.4, 7.1, 7.5 D A Y S T U R N O V E R T I M E , A N D 9.6, 14.0 A N D 13.3$ D A I L Y N E T D E G R A D A T I O N WERE O B S E R V E D F O R L I V E R P R O T E I N S I N C H I C K S C O N S U M I N G 0.36, 0.86 A N D 1.24$ L Y S I N E R E S P E C T I V E L Y . THE R A T E S O F L I V E R P R O T E I N S Y N T H E S I S W E R E 20.0, 49.2 A N D 52.9 M G . / D A Y I N C H I C K S F E D 0.36, 0.86 A N D 1.24$ L Y S I N E R E S P E C T I V E L Y. THE T U R N O V E R R A T E S O F B O T H T H E L I V E R A N D M U S C L E T I S S U E P R O T E I N S W E R E I N C R E A S E D A S T H E L E V E L O F D I E T A R Y L Y S I N E I N C R E A S E D F R O M 0.36 TO 0.86 A N D 1.24$. THE R E S P O N S E C U R V E S O F T H E C H A N G E S I N T U R N O V E R R A T E S O F T I S S U E P R O T E I N S P L O T T E D A G A I N S T T H E L Y S I N E L E V E L WERE S I M I L A R TO T H A T O F B O D Y W E I G H T G A I N P L O T T E D I N A S I M I L A R W AY. R E S U L T S D E M O N S T R A T E T H A T T H E R A T E O F P R O T E I N S Y N T H E S I S A N D D E G R A D A T I O N I S C O R R E L A T E D W I T H T H E C H I C K G R O W T H . T H E R A T E O F P R O T E I N S Y N T H E S I S A N D D E G R A D A T I O N I N T H E L I V E R A N D M U S C L E O F L Y S I N E D E F I C I E N T C H I C K S I S A L S O C O N S I D E R A B L Y L E S S T H A N . I N T H E T I S S U E O F C H I C K S F E D A D E Q U A T E L Y S I N E . TRIAL 5. E F F E C T S O F D I E T A R Y L Y S I N E ON T H E S Y N T H E S I S O F RNA I N T H E L I V E R O F C H I C K S A T E I G H T D A Y S O F A G E A R E S U M M A R I Z E D I N T A B L E 20. T H E A M O U N T O F R A D I O A C T I V I T Y I N L I V E R H O M O G E N A T E WAS NOT S I G N I F I C A N T L Y A F F E C T E D B Y D I E T A R Y L Y S I N E . S I G N I F I C A N T L Y H I G H E R R A D I O A C T I V I T Y I N RNA E X T R A C T (P -<0.01), RNA E X T R A C T E X P R E S S E D A S A P E R C E N T A G E O F T H E R A D I O A C T I V I T Y I N L I V E R H O M O G E N A T E (P «=0.05) A N D S P E C I F I C A C T I V I T Y (P •< 0.01) W E R E , H O W E V E R , O B S E R V E D I N C H I C K S F E D 1.24$ L Y S I N E C O M P A R E D W I T H T H O S E C O N S U M I N G L O W E R L E V E L S O F L Y S I N E . N O S I G N I F I C A N T D I F F E R E N C E I N T H E P A R A M E T E R S WAS O B S E R V E D B E T W E E N T H E 0.36 A N D 0.86$ D I E T A R Y L Y S I N E G R O U P S . R A D I O A C T I V I T Y I N M U S C L E H O M O G E N A T E I N C R E A S E D S I G N I F I C A N T L Y (p < 0.05) B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36 TO 0.86$ ( T A B L E 21). A F U R T H E R I N C R E A S E I N D I E T A R Y L Y S I N E TO 1.24$ R E S U L T E D I N A M U S C L E H O M O G E N A T E A C T I V I T Y V A L U E I N T E R M E D I A T E B E T W E E N T H O S E O F 0.36 A N D 0.86$ L Y S I N E G R O U P S . R A D I O A C T I V I T Y I N RNA E X T R A C T W E R E S L I G H T L Y H I G H E R I N C H I C K S F E D 0.86 A N D 1.24$ L Y S I N E T H A N I N C H I C K S F E D 0.36$ L Y S I N E . R A D I O A C T I V I T Y I N RNA E X P R E S S E D A S A P E R C E N T A G E O F T H E A C T I V I T Y I N M U S C L E H O M O G E N A T E A N D T H E S P E C I F I C A C T I V I T Y W E R E NOT A F F E C T E D TO A N Y S I G N I F I C A N T D E G R E E B Y T H E L Y S I N E T R E A T M E N T S . 1 4 L I V E R D A T A S U G G E S T T H A T T H E I N C O R P O R A T I O N O F C F R O M U R A C I L - 2 -14 U- C I N T O RNA I S I N C R E A S E D B Y G I V I N G T H E C H I C K A D E Q U A T E D I E T A R Y L Y S I N E (1.24$). WHEN L O W E R A M O U N T S O F L Y S I N E A R E F E D , I N T H I S C A S E 0.36 A N D 0.86$, RNA S Y N T H E S I S A S M E A S U R E D B Y T H E C R I T E R I A A L R E A D Y D E S C R I B E D WAS L O W E R E D . C H I C K S U S E D I N T H I S T R I A L U N D O U B T E D L Y C O U L D T O L E R A T E A L E V E L O F 1.24$ D I E T A R Y L Y S I N E A S O B S E R V E D I N T R I A L 4. TABLE 2 0 . - - E F F E C T S O F D I E T A R Y L Y S I N E ON L I V E R RNA S Y N T H E S I S R A D I P A C T I v I T Y  L I V E R RNA RNA E X T R A C T .„2 L Y S I N E T R E A T M E N T {%) H O M O G E N A T E E X T R A C T L I V E R H O M O G E N A T E S P E C I F I C A C T I V I T Y S U P P . L E V E L D P M / G . T I S S U E % D P M / M G . RNA A * A * * A * A * * 0.36 2293.0 507.6 22.2 62.9 0.50 0.86 2492.3 A 515.6 A 20.7 A 62.1 A 0.88 1 .24 2479.1 A 635.0 B 25.7 B 81 .9 B * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P "ZQ.Qfc). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P < 0 . 0 1 ) . TABLE 2 1 . - - E F F E C T O F D I E T A R Y L Y S I N E ON M U S C L E RNA S Y N T H E S I S * RAD I O A C T I V I T Y M U S C L E RNA RNA E X T R A C T 1 „2 — x 1 0 L Y S I N E T R E A T M E N T (%) H O M O G E N A T E E X T R A C T L I V E R H O M O G E N A T E S P E C I F I C A C T I V I T Y S U P P . L E V E L D P M / G . T I S S U E % D P M / M G . RNA 0.36 1462.0 A 326.2 A 22.3 A 181.2 A 0.50 0.86 1646.3 B 347.0 A 21.1 A 168.4 A 0.88 1.24 1580.4 A B 347.4 A 22.0 A 168.3 A * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P <0 . 0 5 ) . 0.86$ D I E T A R Y L Y S I N E WAS O B S E R V E D TO S U P P O R T M A X I M U M W E I G H T G A I N O F C H I C K S A T 14 A N D 28 D A Y S O F A G E I N T R I A L 1. H O W E V E R , I T WAS N O T E D ( T R I A L 1, F I G . 1) T H A T C H I C K S F E D A H I G H (1.11$) L Y S I N E D I E T C O N S U M E D M O R E F E E D F O R T H E F I R S T E I G H T D A Y S O F A G E F O L L O W E D B Y A D E C L I N E I N F E E D I N T A K E C O M P A R E D TO C H I C K S R E C E I V I N G A D E Q U A T E (0.86$) D I E T A R Y L Y S I N E . DATA ON T H E L I V E R I N T H I S T R I A L A N D ON F E E D I N T A K E I N T R I A L 1 ( F I G . 1) S U G G E S T T H A T T H E C H I C K R E Q U I R E M E N T F O R L Y S I N E D E C L I N E S W I T H A G E , A S R E P O R T E D B Y F L S H E R (1972) A N D C H U N G E T A L . (1973), H I G H (1.11 TO 1.36$) F R O M O N E D A Y TO E I G H T D A Y S O F A G E A N D 0.86$ O F T H E D I E T T H E R E A F T E R . T H I S V I E W I S C O N S I S T E N T W I T H T H E H I G H E S T L E V E L O F R A D I O A C T I V E U R A C I L I N C O R P O R A T E D I N T O RNA I N T H E C H I C K S F E D 1.24$ D I E T A R Y L Y S I N E A T E I G H T D A Y S O F A G E C O M P A R E D W I T H T H E C H I C K S F E D 0.36 A N D 0.86$ L Y S I N E A N D C O N F I R M E D B Y T H E H I G H E S T G A I N I N B O D Y W E I G H T , L I V E R W E I G H T A N D T O T A L L I V E R RNA C O N C E N T R A T I O N ( T A B L E 22). T R I A L S 4 A N D 5 WERE R U N S I M U L T A N E O U S L Y A N D C H I C K S M A I N T A I N E D U N D E R T H E S A M E E X P E R I M E N T A L C O N D I T I O N S . L I V E R D A T A I N T H I S T R I A L A G R E E W I T H P R E V I O U S O B S E R V A T I O N S I N T H E L I T E R A T U R E . I N C O R P O R A T I O N O F C - G L Y C I N E I N T O P U R I N E B A S E S A N D 1 4 C - O R O T I C A C I D I N T O T H E P Y R I M I D I N E B A S E S O F R A T L I V E R RNA WAS N O T E D (MUNRO A N D C L A R K , 1959) TO BE C O N S I D E R A B L Y G R E A T E R W H E N A C O M P L E T E A M I N O A C I D M I X T U R E W I T H O U T T R Y P T O P H A N WAS G I V E N TO R A T S W I T H 3 5- H O R O T I C A C I D B Y A S T O M A C H T U B E . H E B E R T _ET _A_L . (1972) B Y A D M I N I S T E R I N G U R I D I N E - 5 - H O R A L L Y W I T H M I X T U R E S O F V A R I O U S A M I N O A C I D S A B O V E T H E C H I C K R E Q U I R E M E N T D E C R E A S E D RNA B I O S Y N T H E S I S . T H E I N C R E A S E D S Y N T H E S I S A N D D E G R A D A T I O N ( T U R N O V E R R A T E ) O F T I S S U E P R O T E I N W I T H I N C R E A S I N G L E V E L O F D I E T A R Y L Y S I N E O B S E R V E D I N TABLE 2 2 .--EFFECT O F D I E T A R Y L Y S I N E ON B O D Y W E I G H T , L I V E R W E I G H T A N D A M O U N T S OF RNA I N C H I C K L I V E R A N D M U S C L E A V E . BODY WT. RNA C O N T E N T  L Y S I N E T R E A T M E N T {%) ( G . ) BODY WT. L I V E R L I V E R M U S C L E S U P P . L E V E L I N I T I A L F I N A L C H A N G E {%) WT. ( G . ) MG./G J M G . / L I V E R M G , / G . 0.36 60 61 1.7 1.8 8.1 14.6 1.8 0.50 0.86 63 69 9.5 2.9 8.3 24.1 2.1 0.88 1.24 77 87 13.0 4.9 7.8 38.2 2.1 DATA R E P R E S E N T M G . / G . WET T I S S U E . T R I A L 4 M I G H T B E C O N S I D E R E D A S A N O C C U R R E N C E A C C O M P A N I E D B Y A N I N C R E A S E D S Y N T H E S I S O F N U C L E I C A C I D (RNA). THIS V I E W I S C O N S I S T E N T W I T H T H E I N C R E A S E D RNA S Y N T H E S I S I N T H E L I V E R O F C H I C K S A S T H E L E V E L O F D I E T A R Y L Y S I N E I N C R E A S E D I N T H E P R E S E N T T R I A L . P R E V I O U S R E P O R T S I N D I C A T E A C O R R E L A T I O N B E T W E E N T H E R E S P O N S E C U R V E O F L I V E R X A N T H I N E O X I D A S E A N D T H A T O F T H E T U R N O V E R R A T E O F T I S S U E P R O T E I N ( M U R A M A T S U A N D A S H I D A , 1962, 1963). X A N T H I N E O X I D A S E P A R T I C I P A T E S I N T H E C A T A B O L I S M O F T H E P U R I N E B A S E S O F N U C L E I C A C I D . AN I N C R E A S E D T U R N O V E R R A T E O F T I S S U E P R O T E I N M I G H T , T H E R E F O R E , B E A C C O M P A N I E D B Y AN. I N C R E A S E D D E G R A D A T I O N O F N U C L E I C A C I D (RNA). T H E C O N S T A N C Y O F P R O T E I N / R N A R A T I O I R R E S P E C T I V E O F T H E L E V E L O F D I E T A R Y L Y S I N E O B S E R V E D I N T R I A L S 1, 3 A N D 4 S U P P O R T S T H E S A M E V I E W , S U M M A R Y A N D C O N C L U S I O N E F F E C T S O F D I E T A R Y L Y S I N E ON L I V E R A N D M U S C L E RNA S Y N T H E S I S W E R E M E A S U R E D . T H R E E D I E T A R Y L Y S I N E L E V E L S (0.36, 0.86, 1.24$) W E R E 14, F E D TO C H I C K S F O R E I G H T D A Y S . ON D A Y S E V E N , U R A C I L - 2 - U - C (60 M-Cl/M M O L E ) WAS I N J E C T E D I N T R A P E R I T O N E A L L Y TO A L L C H I C K S . T H E C H I C K S W E R E S A C R I F I C E D A F T E R 24 H O U R S A N D R A D I O A C T I V I T Y I N C O R P O R A T I O N I N T O L I V E R A N D M U S C L E RNA D E T E R M I N E D . T H E H I G H E S T I N C O R P O R A T I O N O F 14 14 «t C F R O M U R A C I L - 2 - U - C I N T O RNA WAS O B T A I N E D W I T H 1.24$ L Y S I N E I N T H E L I V E R . LOWER A M O U N T S O F L Y S I N E D E C R E A S E D T H E S Y N T H E S I S O F L I V E R RNA. S Y N T H E S I S O F RNA I N T H E M I X E D T I B I A M U S C L E O F C H I C K S F E D 0.86 A N D 1.24$ L Y S I N E S H O W E D A H I G H E R T R E N D T H A N I N C H I C K S F E D 0.36$ L Y S I N E . R E S U L T S D E M O N S T R A T E T H A T T H E M E T A B O L I S M O F RNA C A N B E I N F L U E N C E D B Y D I E T A R Y L Y S I N E ; I N T H I S C A S E RNA S Y N T H E S I S I S D E C R E A S E D B Y I N A D E Q U A T E D I E T A R Y L Y S I N E . TRIAL 6. DATA I N T A B L E 23 R E P R E S E N T A S U M M A R Y O F T H E E F F E C T O F D I E T A R Y A M I N O A C I D T R E A T M E N T ON L Y S I N E O X I D A T I O N I N T H E C H I C K . O X I D A T I O N O F L Y S I N E WAS S I G N I F I C A N T L Y ( P < 0 . 0 5 ) I N C R E A S E D I N C H I C K S F E D E X C E S S L Y S I N E (1.24$) C O M P A R E D W I T H T H O S E C O N S U M I N G A D E Q U A T E L Y S I N E (0.86$). E X C E S S L Y S I N E A L S O D E P R E S S E D C H I C K G R O W T H M A R K E D L Y (P < 0.05). THE D E P R E S S I O N O F C H I C K G R O W T H B Y E X C E S S D I E T A R Y L Y S I N E WAS O B S E R V E D I N T R I A L S 1 A N D 2 A N D P A R T L Y A C C O U N T E D F O R B Y D E P R E S S E D F E E D I N T A K E I N T H I S G R O U P O F C H I C K S . H I G H E R F E E D I N T A K E A N D S I G N I F I C A N T L Y (P < 0.05) I M P R O V E D C H I C K G R O W T H WERE O B T A I N E D W H E N A R G I N I N E WAS A D D E D TO T H E D I E T C O N T A I N I N G E X C E S S L Y S I N E . T H E W H E A T - S O Y B E A N (15$ P R O T E I N ) WAS L I M I T I N G I N L Y S I N E F O R 14 14 14 C H I C K G R O W T H . O X I D A T I O N O F C F R O M L Y S I N E - U - C TO C0^ I N C H I C K S C O N S U M I N G T H E W H E A T - S O Y A D I E T WAS S I G N I F I C A N T L Y (P *= 0.05) L E S S T H A N I N C H I C K S F E D W H E A T P L U S 0.5$ L - L Y S I N E ( A T O T A L O F 0.86$ L Y S I N E ) . S U P P L E M E N T I N G W H E A T - S O Y B E A N D I E T W I T H 0.375$ S U P P L E M E N T A L L Y S I N E S L I G H T L Y I N C R E A S E D L Y S I N E O X I D A T I O N A N D I M P R O V E D F E E D I N T A K E A N D C H I C K G R O W T H S I G N I F I C A N T L Y (P «= 0.05) C O M P A R E D W I T H U N S U P P L E M E N T E D W H E A T -S O Y B E A N D I E T . A F U R T H E R A D D I T I O N O F 0.4$ L - A R G I N I N E TO T H E W H E A T - S O Y B E A N D I E T S U P P L E M E N T E D W I T H 0.375$ L - L Y S I N E R E S U L T E D I N NO S I G N I F I C A N T C H A N G E I N L Y S I N E O X I D A T I O N , F E E D I N T A K E A N D C H I C K G R O W T H C O M P A R E D W I T H T H E W H E A T - S 0 Y B E A N P L U S 0.375$ L - L Y S I N E D I E T . R E S U L T S I N D I C A T E T H A T C H I C K S F E D A L Y S I N E D E F I C I E N T D I E T 14, R E T A I N E D M O R E A N D O X I D I Z E D L E S S O F T H E L Y S I N E - U - C I N J E C T E D 1 N T R A P E R I T O N E A L L Y T H A N C H I C K S R E C E I V I N G A D E Q U A T E A N D H I G H L Y S I N E . R E S U L T S A R E C O N S I S T E N T W I T H T H E S I G N I F I C A N T D E C R E A S E I N P E R C E N T A G E TABLE 23.— P E R C E N T A G E R E C O V E R Y O F R A D I O A C T I V I T Y I N R E S P I R A T O R Y COV, A F T E R I N J E C T I O N OF L Y S I N E - U - C, B O D Y W E I G H T G A I N A N D F E E D I N T A K E O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D W I T H T H R E O N I N E A N D 1 A R G I N I N E ' 2 D I E T A R Y R E S P I R A T O R Y G A I N / C H I C K F E E D I N T A K E / C H I C K T R E A T M E N T C 0 2 ( $ ) ( G . ) ( G . ) W H E A T - B A S A L + 0.5$ L - L Y S I N E W H E A T - B A S A L + 0.875$ L - L Y S I N E W H E A T - B A S A L + 0.875$ L - L Y S I N E + 0.4$ L - A R G I N I N E W H E A T - B A S A L + 0.5$ L - L Y S I N E + 0.16$ L - T H R E O N I N E W H E A T - S O Y B E A N (15$ P R O T E I N ) W H E A T - S O Y B E A N + 0.375$ L - L Y S I N E W H E A T - S O Y B E A N + 0.375$ L - L Y S I N E + 0.4$ L - A R G I N I N E 0.358 469.7°D 1239.2CD 0.59C 254.1A 793.3A 0.91° 355.8B A B 963.9 0.27AB 0.1 oA 0.29AB 612.3E 414.3BC 563.9 1593.4E 1117.6BC 14,04.0DE 0.35B 556.6°E C D 1318.1 V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P "^O.OS) / 14 2 2., D P M / T O T A L C0 o x 10 V A L U E S WERE C A L C U L A T E D A S F O L L O W S : ^ ^ — R E C O V E R Y O F R A D I O A C T I V I T Y A N D S P E C I F I C A C T I V I T Y ( D P M / M G . RNA) I N T H E M U S C L E A N D L I V E R P R O T E I N S O F T H E C H I C K W I T H I N C R E A S I N G D I E T A R Y L Y S I N E (0.36 TO 1.24$) O B S E R V E D I N T R I A L 4. C H I C K S C O N V E R T E D MORE L Y S I N E I N T O T I S S U E ( B O D Y G R O W T H ) W H E N T H E D I E T WAS A D E Q U A T E I N L Y S I N E (0.86$) T H A N W H E N E X C E S S L Y S I N E D I E T WAS F E D . T H R E O N I N E B E C A M E L I M I T I N G WHEN T H E D I E T A R Y L Y S I N E WAS I N C R E A S E D TO 0.86$. T H R O U G H T H E A D D I T I O N O F T H R E O N I N E TO T H E D I E T , F E E D I N T A K E A N D G R O W T H WERE M A R K E D L Y I M P R O V E D A N D L Y S I N E O X I D A T I O N WAS S L I G H T L Y R E D U C E D ( T A B L E 23). MORE L Y S I N E WAS U S E D F O R T I S S U E D E P O S I T I O N W I T H A D E Q U A T E D I E T A R Y L Y S I N E A N D S U P P L E M E N T A L T H R E O N I N E . R E S U L T S I N D I C A T E T H A T T H E M E T A B O L I C P O O L O F L Y S I N E WAS E X C E S S I V E C O M P A R E D IPO O T H E R A M I N O A C I D S A N D C O U L D N O T B E C H A N N E L E D I N T O P R O T E I N S Y N T H E S I S WHEN E X C E S S D I E T A R Y L Y S I N E WAS F E D . T H E S E R E S U L T S A R E S U P P O R T E D B Y L O W E R M U S C L E P R O T E I N / D N A A N D RNA/DNA R A T I O S I N C H I C K S F E D E X C E S S (1.36 A N D 1.86$) L Y S I N E T H A N I N T H O S E R E C E I V I N G A D E Q U A T E (0.86$) L Y S I N E O B S E R V E D I N T R I A L 2. T H E R E F O R E , E X C E S S L Y S I N E I N T H E C H I C K WAS E L I M I N A T E D B Y O X I D A T I O N . F E E D I N T A K E WAS D R A S T I C A L L Y R E D U C E D U N D E R T H E C O N D I T I O N O F E X C E S S L Y S I N E I N A N A T T E M P T B Y T H E C H I C K TO A D J U S T T H E M E T A B O L I C A M I N O A C I D P O O L T H A T S U G G E S T S A P R O T E C T I V E M E C H A N I S M . A R G I N I N E S U P P L E M E N T A T I O N I N C R E A S E D T H E C A P A C I T Y O F T H E C H I C K TO I N C O R P O R A T E M O R E L Y S I N E I N T O B O D Y P R O T E I N S A N D O X I D I Z E E X C E S S L Y S I N E . R E S U L T S A G R E E W I T H T H E R E P O R T O F B R O O K E S _ET _ A L . (1972) T H A T O X I D A T I O N O F L Y S I N E D I D N O T I N C R E A S E M A R K E D L Y U N T I L T H E D I E T A R Y L Y S I N E I N T A K E WAS I N C R E A S E D A B O V E T H E L E V E L A T W H I C H A V E R A G E D A I L Y G A I N A N D G A I N / F E E D W E R E M A X I M A L . IT WAS A L S O D E M O N S T R A T E D ( Y A M A S H I T A A N D A S H I D A , 1969) T H A T T H E P E R C E N T A G E R E C O V E R Y O F C F R O M 14, L Y S I N E - U - C I N R E S P I R A T O R Y C 0 2 I N R A T S F E D L Y S I N E - F RE E D I E T WAS L E S S T H A N I N R A T S C O N S U M I N G A D E Q U A T E L Y S I N E . IT H A S B E E N E S T A B L I S H E D T H A T E X C E S S D I E T A R Y L Y S I N E I S T O X I C TO T H E C H I C K A N D T H E A D V E R S E - E F F E C T C A N B E A L L E V I A T E D B Y A R G I N I N E S U P P L E M E N T A T I O N ( T R I A L 2; J O N E S , 1964] S M I T H A N D L E W I S , 1966; O'DELL A N D S A V A G E , 1966; S M I T H , 1968; A U S T I C A N D N E S H E I M , 1970). A D D I T I O N O F 0.2$ D L - T H R E O N I N E TO W H O L E W H E A T D I E T S U P P L E M E N T E D W I T H 0.25$ L - L Y S I N E HCIL A N D 0.5$ D L - V A L I N E WAS A L S O S H O W N TO I M P R O V E T H E R A T G R O W T H ( S U R E , 1952). S U M M A R Y A N D C O N C L U S I O N E F F E C T S O F D I E T A R Y A M I N O A C I D L E V E L I N T H E C H I C K D I E T ON L Y S I N E O X I D A T I O N WAS S T U D I E D . C H I C K S F E D D I F F E R E N T L E V E L S O F L Y S I N E , A D E Q U A T E L Y S I N E W I T H T H R E O N I N E A N D E X C E S S L Y S I N E W I T H A R G I N I N E WERE 14 R I N J E C T E D W I T H L Y S I N E - U - C I N T R A P E R I T O N E A L L Y . T H E P R O D U C T I O N O F R A D I O A C T I V E CC^ ( R E C O V E R Y A S A P E R C E N T A G E O F D O S E ) WAS U S E D A S A N I N D E X O F A M O U N T O F L Y S I N E O X I D I Z E D . R E S U L T S I N D I C A T E D T H A T T H E O X I D A T I O N O F L Y S I N E I N C R E A S E D W I T H D I E T A R Y L Y S I N E L E V E L . T H R E O N I N E S U P P L E M E N T A T I O N T O T H E L Y S I N E A D E Q U A T E D I E T R E D U C E D L Y S I N E O X I D A T I O N S L I G H T L Y A N D I M P R O V E D C H I C K G R O W T H A N D F E E D I N T A K E M A R K E D L Y . E X C E S S L Y S I N E D E P R E S S E D C H I C K G R O W T H A N D F E E D I N T A K E A N D T H E A D V E R S E - E F F E C T S O F E X C E S S L Y S I N E W E R E A L L E V I A T E D B Y A R G I N I N E S U P P L E M E N T A T I O N . A N I N C R E A S E I N D I E T A R Y A R G I N I N E A L S O I N C R E A S E D T H E C A P A C I T Y O F T H E C H I C K TO OX I D I Z E L Y S I N E . 143 TRIAL 7. R E S U L T S O F T R I A L 6 I N D I C A T E T H A T T H E O X I D A T I O N O F L Y S I N E I N T H E C H I C K I N C R E A S E D A S T H E L E V E L O F D I E T A R Y L Y S I N E I N C R E A S E D . T H I S T R I A L WAS C O N D U C T E D TO D E T E R M I N E T H E P A T T E R N O F L Y S I N E I N C O R P O R A T I O N I N T O T I S S U E P R O T E I N S U N D E R I N C R E A S I N G D I E T A R Y L Y S I N E R E G I M E S . M O R E O V E R , T H R E O N I N E S U P P L E M E N T A T I O N TO T H E L Y S I N E A D E Q U A T E D I E T R E D U C E D L Y S I N E O X I D A T I O N S L I G H T L Y A N D I M P R O V E D F E E D I N T A K E A N D C H I C K G R O W T H M A R K E D L Y I N T R I A L 6. THE M E T A B O L I C F A T E O F T H R E O N I N E I N C H I C K S F E D D I E T S C O N T A I N I N G A D E F I C I E N T , A D E Q U A T E A N D E X C E S S L E V E L O F L Y S I N E W A S , T H E R E F O R E , I N V E S T I G A T E D I N T H I S T R I A L . E F F E C T S O F T H E L E V E L O F D I E T A R Y L Y S I N E ON I N C O R P O R A T I O N O F L Y S I N E - U - C A N D T H R E O N I N E - U - C I N T O C H I C K L I V E R A N D M U S C L E P R O T E I N S A R E I L L U S T R A T E D I N T A B L E S 24 A N D 25 R E S P E C T I V E L Y . THE 1 4 S P E C I F I C A C T I V I T Y V A L U E S O F I N J E C T E D L Y S I N E - U - C I N TCA - I N S O L U B L E P R O T E I N S I N T H E L I V E R A N D M U S C L E S H O W E D A D E C L I N I N G T R E N D W I T H I N C R E A S I N G L E V E L O F D I E T A R Y L Y S I N E ( T A B L E 24). I N C R E A S I N G L Y S I N E L E V E L I N T H E C H I C K D I E T R E S U L T E D I N A N E L E V A T E D R A D I O A C T I V I T Y R E C O V E R E D I N T C A - S O L U B L E P R O T E I N S I N B O T H T H E L I V E R A N D M U S C L E , T H E I N C R E A S E I N R A D I O A C T I V I T Y WAS S I G N I F I C A N T L Y (P -=0.01) H I G H E R W I T H 1.24$ L Y S I N E . A C H A N G E I N T H E R E C O V E R Y O F R A D I O A C T I V I T Y I N P R O T E I N F R A C T I O N S D O E S NOT N E C E S S A R I L Y R E F L E C T A N E N H A N C E D S Y N T H E S I S O F P R O T E I N M O L E C U L E S , S I N C E T H E P O O L S I Z E O F A M I N O A C I D M A Y R E A D I L Y I N F L U E N C E T H E I N C O R P O R A T I O N O F R A D I O A C T I V E A M I N O A C I D S I N T O P R O T E I N . T H E R E L A T I V E S P E C I F I C A C T I V I T Y ( T C A - I N S O L U B L E P R O T E I N S P E C I F I C A C T I V I T Y / T C A - S O L U B L E P R O T E I N S P E C I F I C A C T I V I T Y ) W A S , T H E R E F O R E , C A L C U L A T E D TO D E T E R M I N E T H E E X T E N T T H A T A M I N O A C I D S WERE R E T A I N E D I N TABLE 2 4 . - - P E R C E N T A G E R E C O V E R Y O F R A D I O A C T I V I T Y I N C H I C K L I V E R A N D M U S C L E P R O T E I N A F T E R I N J E C T I O N O F L Y S I N E - U - 1 4C L I V E R M U S C L E * L Y S I N E T R E A T M E N T (%) A* B** C* A B C S U P P . L E V E L fo i 0.36 4.99A 0.23A 21.64A 2.92A 0.1S55A 19. A 12 0.50 0.86 4.1 6A 0.25A 16.648 2.61 A 0.243A 10. 75A 0.88 1 .24 3.84A 0.32B 11.99° 2.52A 0.4338 5. 85A A = T I S S U E TCA 1 N S O L U B L E P R O T E I N S P E C I F I C A C T 1V 1 T Y D P M / G . P R O T E I N x 102 1.11 x 10° • D T T O A D P M / G . T I S S U E .. -2 B = T I S S U E TCA S O L U B L E P R O T E I N S P E C I F I C A C T I V I T Y -—r\— .nu x 10 . 1.11 x 10° . o D P M / G . P R O T E I N C = R E L A T I V E S P E C I F I C A C T I V I T Y (A/B) h : . X ' ' -DPM/G. T I S S U E * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P <0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P "=0.01). TABLE 2 5 . - - P E R C E N T A G E R E C O V E R Y O F R A D I O A C T I V I T Y I N C H I C K L I V E R A N D M U S C L E P R O T E I N A F T E R I N J E C T I O N O F T H R E O N I N E - U -L 1 V E R * * M U S C L E L Y S I N E T R E A T M E N T (%) A B c A B C S U P P . L E V E L % % 0.36 2.53 A 0.25 A 10.39 A A * * 1 .43 A * * 0.139 10. A * 36 0.50 0.86 3/05 B 0.12 B 25.92 B 1 .59 B 0.082 8 19. 89 B 0.88 1.24 2.75° 0.11 B 25.76 B 1 .92° 0.073 6 26. 5 8 B A, B A N D C R E P R E S E N T A C T I V I T Y C A L C U L A T E D A S I N T A B L E 2%. * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S 1 G N 1 F 1 C A N T L Y D 1 F F E RE NT (P «= 0.05). * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D 1 F F E R E N T ( P < 0.01). T H E T I S S U E P O O L I N T H E B O D Y OR E X C R E T E D . R E S U L T S S H O W E D T H A T T H E R A T I O D E C R E A S E D S I G N I F I C A N T L Y (P <0.05) W I T H I N C R E A S I N G D I E T A R Y L Y S I N E (.36 TO 1.24$) I N T H E L I V E R A N D N O N - S I G N I F I C A N T L Y I N T H E M U S C L E . A C C U M U L A T E D R E S U L T S I N T H I S T R I A L A N D T R I A L 4 S T R O N G L Y I N D I C A T E T H A T T H E R E T E N T I O N O F L Y S I N E I N T H E C H I C K D E C R E A S E D W I T H I N C R E A S I N G D I E T A R Y L Y S I N E I N T A K E . S I N C E O X I D A T I O N O F L Y S I N E I N C R E A S E D W I T H D I E T A R Y L Y S I N E ( T R I A L 6 ) , T H E P R E S E N T T R I A L WAS C O N D U C T E D TO I N V E S T I G A T E T H E M E T A B O L I C F A T E O F D I E T A R Y L Y S I N E . L Y S I N E R E T E N T I O N I N T H E C H I C K D E C L I N E D W I T H I N C R E A S I N G D I E T A R Y L Y S I N E C O N T E N T A S S H O W N I N T A B L E 24. THE D E C R E A S E I N R A T E O F D E P O S I T I O N WAS C O N C U R R E N T W I T H A N I N C R E A S E I N O X I D A T I O N T H A T R E S U L T E D F R O M F E E D I N G E X C E S S L Y S I N E 1 4 ( T R I A L 6 ) . THE R A T E O F L Y S I N E - U - C D E P O S I T I O N A P P E A R E D TO B E R E L A T E D TO T H E D E M A N D F O R L Y S I N E F O R P R O T E I N S Y N T H E S I S . D E F I C I E N T C H I C K S , (.36$ L Y S I N E ) HAD T H E H I G H E S T S P E C I F I C A C T I V I T Y T H A T D E C R E A S E D W I T H 0.86$ A N D E X C E S S L Y S I N E (1.24$). THE T H R E E L Y S I N E L E V E L S , 0.36, 0.86 A N D 1.24$, R E S P E C T I V E L Y S U P P O R T E D 61, 94 A N D 84 G. W E I G H T G A I N I N S I X D A Y S , I N P R O P O R T I O N TO 183, 229 A N D 194 G. F E E D I N T A K E T H A T F U R T H E R I N D I C A T E T H A T L Y S I N E R E T E N T I O N WAS I N F L U E N C E D B Y T H E D E M A N D F O R P R O T E I N S Y N T H E S I S . R E S U L T S O F T H E P R E S E N T T R I A L , A R E I N A G R E E M E N T W I T H R E P O R T S ( Y A M A S H I T A A N D A S H I D A , 1969, 1971) T H A T R E T E N T I O N O F L Y S I N E I N T H E L I V E R S O F R A T S F E D A L Y S I N E - F R E E D I E T WAS M U C H H I G H E R T H A N T H A T I N R A T S R E C E I V I N G A D E Q U A T E L Y S I N E . T H E F E E D I N G O F E X C E S S L Y S I N E TO R A T S WAS A L S O R E P O R T E D TO R E S U L T I N A L O W E R R A D I O A C T I V I T Y R E C O V E R Y I N T H E P R O T E I N F R A C T I O N I N C O N T R A S T TO T H E E L E V A T E D R E C O V E R Y I N T H E A C I D - S O L U B L E F R A C T I O N . 14, R E T E N T I O N O F T H R E O N I N E - U - C I N T H E C H I C K WAS I N F L U E N C E D B Y D I E T A R Y L Y S I N E C O N T E N T ( T A B L E 25). R A D I O A C T I V I T Y R E C O V E R Y I N L I V E R T C A - I N S O L U B L E P R O T E I N S WAS S I G N I F I C A N T L Y (P < 0.01) R E D U C E D I N C H I C K S F E D D I E T S W I T H L I M I T E D A N D E X C E S S L Y S I N E C O M P A R E D W I T H T H E G R O U P 14 R E C E I V I N G A D E Q U A T E L Y S I N E . T H R E O N I N E - U - C R E C O V E R Y I N T C A - I N S O L U B L E P R O T E I N S WAS I N D I R E C T C O N T R A S T W I T H T H E S I G N I F I C A N T L Y (P < 0.01) G R E A T E R R A D I O A C T I V I T Y ( T H R E 0 N I N E - U - ^ C ) R E C O V E R E D I N L I V E R T C A - S O L U B L E P R O T E I N S O F L Y S I N E D E F I C I E N T C H I C K S T H A N I N T H O S E O F C H I C K S R E C E I V I N G H I G H E R L Y S I N E T R E A T M E N T S . A S I G N I F I C A N T L Y (P <0.01) L O W E R R E L A T I V E S P E C I F I C A C T I V I T Y WAS O B S E R V E D I N L Y S I N E D E F I C I E N T G R O U P S T H A N I N C H I C K S F E D A D E Q U A T E OR E X C E S S I V E L Y S I N E A S A R E S U L T O F T H E D I F F E R E N C E I N R E S P O N S E O F A C I D - I N S O L U B L E A N D A C I D - S O L U B L E T I S S U E P R O T E I N S TO D I E T A R Y L Y S I N E . T H E L E V E L O F R A D I O A C T I V I T Y ( T H R E 0 N I N E - U - C) R E C O V E R E D I N T H E C H I C K M U S C L E E S S E N T I A L L Y R E S E M B L E D T H A T I N T H E L I V E R . R E S U L T S D E M O N S T R A T E T H A T T H E R E T E N T I O N O F T H R E O N I N E I N D I E T A R Y L Y S I N E D E F I C I E N C Y WAS L E S S T H A N WAS O B S E R V E D I N C H I C K S R E C E I V I N G 14 14 A D E Q U A T E L Y S I N E . LOW R E T E N T I O N O F C F R O M T H R E O N I N E - U - C I N T H E C H I C K F E D L Y S I N E D E F I C I E N T D I E T WAS P R O B A B L Y N O R M A L S I N C E L Y S I N E WAS T H E F I R S T L I M I T I N G A M I N O A C I D F O R G R O W T H ( T R I A L S 1 A N D 3 ) . T H R E O N I N E R E T E N T I O N I N T H E L Y S I N E D E F I C I E N T G R O U P WAS NOT N E C E S S A R Y B E C A U S E G R O W T H A N D T I S S U E P R O T E I N S Y N T H E S I S ( T R I A L S 1, 3, 4) W E R E A T L O W E R R A T E S R E Q U I R I N G A L O W E R L E V E L O F D I E T A R Y T H R E O N I N E C O M P A R E D TO C H I C K S R E C E I V I N G A D E Q U A T E L Y S I N E . T H R E O N I N E B E C A M E F I R S T L I M I T I N G W H E N T H E C H I C K D I E T WAS A D E Q U A T E I N L Y S I N E S H O W N B Y T H E I M P A C T O F S U P P L E M E N T A L T H R E O N I N E TO T H E D I E T T H A T I M P R O V E D C H I C K G R O W T H M A R K E D L Y A N D D E C R E A S E D L Y S I N E O X I D A T I O N S L I G H T L Y ( T R I A L 6). M O R E T H R E O N I N E U N D O U B T E D L Y WAS I N C O R P O R A T E D I N T O T I S S U E P R O T E I N S TO A C C O U N T F O R T H E I M P R O V E D G R O W T H I N T R I A L 6 A N D I N A G R E E M E N T W I T H T H E R E S U L T O F T H E P R E S E N T T R I A L . M O R E O V E R , Z I M M E R M A N A N D S C O T T (1965) H A V E D E M O N S T R A T E D T H A T S E V E R E D E F I C I E N C Y O F L Y S I N E I N C R E A S E D P L A S M A T H R E O N I N E C O N C E N -T R A T I O N F R O M A B O U T 40 TO 190 U G . / M L . I N T H E C H I C K . I T WAS A L S O S H O W N T H A T R A T S D E P R I V E D O F L Y S I N E E X C R E T E D MORE T H R E O N I N E I N T H E U R I N E . THE A M O U N T O F E X C R E T E D T H R E O N I N E WAS D E P E N D E N T ON T H E D I E T A R Y L Y S I N E L E V E L ( Y A M A S H I T A A N D S A K U R A I , 1966). R E S U L T S I N T H E P R E S E N T S T U D Y A R E I N A G R E E M E N T W I T H D A T A P R E V I O U S L Y R E P O R T E D ( Z I M M E R M A N A N D S C O T T , 1965J Y A M A S H I T A A N D S A K U R A I , 1966). S U M M A R Y A N D C O N C L U S I O N THE I N F L U E N C E O F D I E T A R Y L Y S I N E C O N T E N T ON L Y S I N E A N D T H R E O N I N E R E T E N T I O N I N T H E L I V E R A N D M U S C L E P R O T E I N S O F T H E C H I C K WAS S T U D I E D . C H I C K S F E D T H R E E L E V E L S (0.36, 0.86, 1.24$) O F L Y S I N E W E R E I N J E C T E D 14, 14, W I T H L Y S I N E - U - C OR T H R E O N I N E - U - C I N T R A P E R I T O N E A L L Y . R A D I O A C T I V I T Y R E C O V E R Y I N L I V E R A N D M U S C L E P R O T E I N S WERE D E T E R M I N E D . R E S U L T S I N D I C A T E T H A T T H E C E L L U L A R L E V E L O F T H E A M I N O A C I D S I S I N F L U E N C E D B Y T H E L E V E L O F D I E T A R Y L Y S I N E . L Y S I N E R E T E N T I O N I N L I V E R A N D M U S C L E P R O T E I N S D E C L I N E D W I T H I N C R E A S I N G D I E T A R Y L Y S I N E . L Y S I N E D E F I C I E N C Y D R A S T I C A L L Y D E C R E A S E D T H R E O N I N E R E T E N T I O N I N B O T H T H E L I V E R A N D M U S C L E . E X C E S S D I E T A R Y L Y S I N E S L I G H T L Y I M P R O V E D T H R E O N I N E R E T E N T I O N I N T H E M U S C L E B U T N O T I N T H E L I V E R . TRIAL 8. R E S U L T S O F T R I A L S 6 A N D 7 I N D I C A T E T H A T T H E O X I D A T I O N A N D R E T E N T I O N O F L Y S I N E I N T I S S U E P R O T E I N O F T H E C H I C K WERE M A R K E D L Y A F F E C T E D B Y D I E T A R Y L Y S I N E C O N T E N T . T H R E O N I N E R E T E N T I O N WAS A L S O S H O W N TO B E D R A S T I C A L L Y I M P A I R E D B Y L Y S I N E D E F I C I E N C Y . T H E P R E S E N T T R I A L WAS C O N D U C T E D TO C O N F I R M T H E S E O B S E R V A T I O N S . R E S U L T S O F T H E R A D I O A C T I V I T Y M E A S U R E M E N T S I N ( A ) R E S P I R A T O R Y CtV,, ( B ) L I V E R P R O T E I N , ( C ) A C I D - S O L U B L E F R A C T I O N I N L I V E R A N D M U S C L E P R O T E I N A N D ( D ) A C I D - S O L U B L E F R A C T I O N I N M U S C L E C O N T A I N I N G I N C O R P O R A T E D 14. L Y S I N E - U - C I N C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A R E P R E S E N T E D I N 1 4 T A B L E 26. R A D I O A C T I V I T Y R E C O V E R Y I N CO^  F R O M L Y S I N E - U - C I N C R E A S E D S I G N I F I C A N T L Y W I T H T H E A M O U N T O F L Y S I N E I N C H I C K D I E T . T H E R A D I O -A C T I V I T Y R E C O V E R E D I N T H E P R O T E I N F R A C T I O N O F B O T H L I V E R A N D M U S C L E D E C R E A S E D W I T H D I E T A R Y L Y S I N E L E V E L W I T H T H E E X C E P T I O N T H A T R E C O V E R Y I N M U S C L E P R O T E I N WAS H I G H E R I N C H I C K S F E D E X C E S S L Y S I N E T H A N I N T H O S E C O N S U M I N G A D E Q U A T E L Y S I N E . R A D I O A C T I V I T Y R E C O V E R Y I N T H E A C I D S O L U B L E F R A C T I O N ( A M I N O A C I D P O O L ) I N B O T H L I V E R A N D M U S C L E I N C R E A S E D M A R K E D L Y W I T H D I E T A R Y L Y S I N E . A D E C L I N E O F R A D I O A C T I V I T Y WAS O B S E R V E D I N L I V E R A N D M U S C L E P R O T E I N S R E L A T I V E TO T H A T O F T H E A C I D - S O L U B L E F R A C T I O N S A S T H E L Y S I N E L E V E L I N T H E C H I C K D I E T I N C R E A S E D . DATA A G R E E W I T H A N D C O N F I R M R E S U L T S O F T R I A L S 6 A N D 7. Low 14, 14, O X I D A T I O N W I T H A H I G H R E T E N T I O N O F C F R O M L Y S I N E - U - C M O N O H Y D R O -C H L O R I D E I N C H I C K S R E C E I V I N G 0.36$ L Y S I N E S U G G E S T S T H E F O L L O W I N G C O N C L U S I O N S : (1) T H E C H I C K I N C O R P O R A T E D A G R E A T A M O U N T O F T H E 14-L Y S I N E - U - C I N T O T I S S U E P R O T E I N T H A T W E R E S L O W L Y M E T A B O L I Z E D A S O B S E R V E D I N T R I A L 4. G R E A T E R R E T E N T I O N P R O B A B L Y WAS N E C E S S A R Y S I N C E TABLE 26.-- P E R C E N T A G E R E C O V E R Y O F R A D I O A C T I V I T Y I N R E S P I R A T O R Y C0 o , L I V E R A N D M U S C L E P R O T E I N A F T E R 14 2 I N J E C T I O N O F L Y S I N E - U - C RES P I R A T O R Y 1 L I V E R M U S C L E ^ L Y S I N E T R E A T M E N T (%) COo A B c A B C S U P P . L E V E L ft fo % A* A * * A * A * A * A** A * 0.36 0.22 6.81 0.37 1 8.40 2.23 0.19 11 .57 0.50 0.86 0.80 A B 3.85 B 0.50 B 7.75 B 1 .59 B 0.55 B 2 .92 B 0.88 1.24 1 .97 B 3.35° 0.74° 4.51° 1 .81° 0.90° 2 .01° 1 R E S P 1 R A T O R Y CO^, k, B A N D C R E P R E S E N T A C T 1V1TY C A L C U L A T E D A S I N T A B L E S 23 A N D 24. * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D 1 F F E R E N T (P < 0.05) • * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S 1 G N I F 1 C A N T L Y D 1 F F E R E N T (P < 0.01 T H E LOW L E V E L O F L Y S I N E I N T H E D I E T WAS I N A D E Q U A T E TO M E E T N E E D S F O R E F F I C I E N T G R O W T H A S D E M O N S T R A T E D B Y T H E P E R F O R M A N C E D A T A ( T A B L E 28). 1 4o R E T E N T I O N O F A C T I V I T Y F R O M L Y S I N E - U - C I N T I S S U E P R O T E I N S O F C H I C K S F E D A D E Q U A T E L Y S I N E WAS LOW S I N C E T H E T U R N O V E R R A T E O F T I S S U E P R O T E I N S WAS G R E A T E R ( T R I A L 4) T H A N I N C H I C K S R E C E I V I N G T H E L Y S I N E D E F I C I E N T D I E T . IN R A D I O A C T I V E S T U D I E S , I T I S B E L I E V E D T H A T A N A M I N O A C I D I S R E L E A S E D OR E X C H A N G E D F R O M W I T H I N A P R O T E I N M O L E C U L E B Y A C O M P L E T E OR P A R T I A L P R O T E I N D E G R A D A T I O N ( T A R V E R , 1954; S T E I N B E R G _ET _AL • 1956). T H E I M P L I C A T I O N I S T H A T A S I N G L E I S O T O P I C A L L Y L A B E L L E D A M I N O A C I D S P E C I E S W I T H I N A P R O T E I N H A S A H A L F - L I F E E Q U A L TO T H A T O F A L L T H E O T H E R A M I N O A C I D S P E C I E S W I T H I N T H E P R O T E I N A N D H E N C E O F T H E P R O T E I N 14„ I T S E L F . T H E I N J E C T E D L Y S I N E - U - C C O U L D , T H E R E F O R E , B E E X C H A N G E D F O R D I E T A R Y L Y S I N E . (2) C H I C K S R E C E I V I N G 0.86 A N D 1.24$ D I E T A R Y L Y S I N E I N C O R P O R A T E D A S M A L L A M O U N T O F T H E I N J E C T E D L Y S I N E - U - C B E C A U S E O F A G R E A T E R D I L U T I O N O F T H E R A D I O A C T I V E A M I N O A C I D B Y I N J E C T E D L Y S I N E . G R E A T E R A C I D S O L U B L E P R O T E I N ( A M I N O A C I D P O O L ) I N T H E S E G R O U P S O F C H I C K S T H A N T H E G R O U P F E D 0.36$ L Y S I N E S U P P O R T S T H E D I L U T I O N T H E O R Y . (3) E X C E S S I V E D I E T A R Y L Y S I N E WAS A M E T A B O L I C B U R D E N T H A T D E P R E S S E D C H I C K G R O W T H M A R K E D L Y ( T A B L E 28). UNDER T H E S E C O N D I T I O N S , E L I M I N A T I O N O F E X C E S S L Y S I N E V I A T H E O X I D A T I O N P A T H W A Y WAS A U G M E N T E D . A U S T I C A N D N E S H E I M (1970) H A V E D E M O N S T R A T E D M A R K E D I N C R E A S E S I N K I D N E Y A R G I N A S E A C T I V I T Y A N D U R E A E X C R E T I O N I N C H I C K S R E C E I V I N G E X C E S S D I E T A R Y L Y S I N E . L Y S I N E I S U N I Q U E I N T H E G E N E R A L S C H E M E O F A M I N O A C I D M E T A B O L I S M B E C A U S E O F I T S R E L A T I V E M E T A B O L I C I N E R T N E S S . L Y S I N E A P P A R E N T L Y D O E S NOT P A R T I C I P A T E I N R E V E R S I B L E T R A N S A M I N A T I O N R E A C T I O N S I N T H E B O D Y ( M E I S T E R , 1957). FROM T H E WORK O F V A N S L Y K E A N D S I N E X (1958), I T WAS A P P A R E N T T H A T L Y S I N E C A N B E C O N V E R T E D TO H Y D R O X Y L Y S I N E , A N A M I N O A C I D F O U N D O N L Y I N C O L L A G E N . G R E E N B E R G A N D R O T H S T E I N (1959) R E P O R T E D T H A T L Y S I N E WAS I R R E V E R S I B L Y T R A N S F O R M E D I N T H E B O D Y V I A P I P I C O L I C A C I D A N D A L P H A - A M I N O A D I P I C A C I D T O G L U T A R I C A C I D . W A N G A N D N E S H E I M (1972) R E C E N T L Y O B S E R V E D T H A T T H E S A C R O P I N E P A T H W A Y I S P R O B A B L Y A N A L T E R N A T I V E P A T H W A Y F O R L - L Y S I N E • D EG R A D A T I O N I N V I V O I N T H E C H I C K TO G L U T A R I C A C I D . T H E G L U T A R I C A C I D M A Y B E O X I D I Z E D T O A L P H A - K E T O G L U T A R I C A C I D A N D T H E N E N T E R T H E T R I C A R B O X Y L I C A C I D (TCA) C Y C L E . T H E N I T R O G E N L O S T B Y S U C H I R R E V E R S I B L E D E A M I N A T I O N P R E S U M A B L Y C A N E N T E R T H E B O D Y P O O L O F N I T R O G E N A N D B E U S E D I N P A R T F O R T H E S Y N T H E S I S O F N O N - E S S E N T I A L A M I N O A C I D S . O N E M O L E C U L E O F C A R B O N D I O X I D E I S U S U A L L Y L I B E R A T E D F R O M O N E M O L E C U L E O F A L P H A - K E T O G L U T A R I C A C I D ON C O N V E R S I O N TO S U C C I N Y L 14 CoA I N T H E TCA C Y C L E . D E G R A D A T I O N O F L Y S I N E - U - C I N J E C T E D I N T O C H I C K S I N T H E P R E S E N T S T U D Y WAS P R O B A B L Y T H R O U G H T H E M E C H A N I S M D E S C R I B E D . T H E R E I S E V I D E N C E , H O W E V E R , T H A T L Y S I N E C A N B E D E G R A D E D TO ACETYL-COA ( BE ND I T T _ET _AL . 1950). 14, , . R E S U L T S O F T H E T R A N S F O R M A T I O N O F T H R E O N I N E - U - C I N T O ( A ) R E S P I R A T O R Y CO^, ( B ) L I V E R P R O T E I N , (c) A C I D - S O L U B L E F R A C T I O N I N L I V E R A N D M U S C L E P R O T E I N A N D ( D ) A C I D - S O L U B L E F R A C T I O N I N M U S C L E O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A R E P R E S E N T E D I N T A B L E 27. D A T A S H O W E D T H A T C H I C K S F E D D I E T S L I M I T E D I N L Y S I N E , O X I D I Z E D T H R E O N I N E M O R E R E A D I L Y T H A N C H I C K S R E C E I V I N G A D E Q U A T E L Y S I N E . T H E O X I D A T I O N O F T H R E O N I N E B Y C H I C K S F E D E X C E S S L Y S I N E (l.24$) WAS I N T E R M E D I A T E B E T W E E N T H E G R O U P S O F C H I C K S F E D 0.36 A N D 0.86$ L Y S I N E . R E S U L T S C O N F I R M T H E O B S E R V A T I O N I N T R I A L . 7 T H A T I N L Y S I N E D E F I C I E N C Y , T H R E O N I N E O X I D A T I O N I S H I G H E R T H A N N O R M A L . TABLE 2 7 . - - P E R C E N T A G E R E C O V E R Y O F R A D I O A C T I V I T Y I N R E S P I R A T O R Y C0 o , L I V E R A N D M U S C L E P R O T E I N A F T E R 1 A 2 I N J E C T I O N O F T H R E O N I N E - U - C 1 R E S P 1 R A T O R Y 1 L I V E R M U S C L E ^ L Y S I N E T R E A T M E N T {%) A B c A B C S U P P . L E V E L r % % 0.36 A * 0.99 A * * 4.08 A * 0.56 7 A * * .32 A * * 1 .73 A * 0.50 A * * 3.44 0.50 0.86 0.24 8 9.78 B 0.26 B 37 .86 B 3.31 8 0.20 8 16.93 S 0.88 1 .24 0.38 A B 4.78° 0.29° 16 .36 C 1 .78 A 0.34 C 5.35 A ^ R E S P I R A T O R Y CO^, A, B A N D C R E P R E S E N T A C T 1V 1 T Y C A L C U L A T E D AS I N T A B L E S 22 A N D 23. * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P < 0.05). * " * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P «=0.01). TABLE 2 8 .--EFFECT O F D I E T A R Y L Y S I N E L E V E L ON T H R E O N I N E U T I L I Z A T I O N B Y T H E C H I C K A M I N O A C I D S U P P L E M E N T TO W H E A T - B A S A L D I E T G A I N / C H I C K G.** F E E D I N T A K E / C H I C K G.** NONE 69.3A 293A + 0.084$ L - T H R E O N 1 N E 76.3A 295A + 0.1 68$ L - T H R E O N 1 N E 68.3A 331.3B 338.3B 364.3B 305A 862 8 9208 954 8 + O.E )$ L- L YS 1 N E + O.E )$ L- L Y S I N E + 0.084$ L- T H R E O N 1 N E + O.E 3$ L-L Y S I N E + 0.168$ L- T H R E O N 1 N E + OA 375$ L - L Y S 1 N E 211.3C 621 C + OA 575$ L - L Y S 1 N E + 0.084$ L - T H R E O N 1 N E 246.0C 652C + O.c 575$ L - L Y S I N E + 0.168$ L - T H R E O N 1 N E 226.0C 374.0B 612C + 0.1 )$ L- L Y S I N E + 0.168$ L-+ 0.15$ DL-T H R E O N 1 N E M E T H 1 ON 1 NE 988B * * V A L U E S F O L L O W E D B Y A D I F F E R E N T S U P E R S C R I P T A R E S I G N I F I C A N T L Y D I F F E R E N T (P *=0.01). R A D I O A C T I V I T Y O F T H R E O N I N E I N C R E A S E D S I G N I F I C A N T L Y (P <= 0.01) I N T H E A C I D- I N S O L U B L E P R O T E I N F R A C T I O N A N D D E C R E A S E D (P < 0.05) I N T H E A C I D - S O L U B L E F R A C T I O N S O F T H E L I V E R A N D M U S C L E W I T H I N C R E A S I N G A M O U N T S O F L Y S I N E I N T H E C H I C K D I E T F R O M 0.36 TO 0.86$. A F U R T H E R I N C R E A S E O F D I E T A R Y L Y S I N E TO 1.24$ D E C R E A S E D T H E R A D I O A C T I V I T Y R E C O V E R E D I N T H E L I V E R A N D M U S C L E P R O T E I N A N D I N C R E A S E D T H E R A D I O A C T I V I T Y I N T H E A C I D - S O L U B L E F R A C T I O N ( A M I N O A C I D P O O L ) . R A D I O A C T I V I T Y O F T H R E O N I N E I N T H E A C I D- I N S O L U B L E P R O T E I N E X P R E S S E D R E L A T I V E TO T H E R E C O V E R Y I N T H E A C I D - S O L U B L E F R A C T I O N I N T H E L I V E R A N D M U S C L E S H O W E D T H E S A M E P A T T E R N . I T I N C R E A S E D S I G N I F I C A N T L Y ( P < 0.01) B Y I N C R E A S I N G D I E T A R Y L Y S I N E F R O M 0.36 A N D .0.86$ A N D D E C R E A S E D TO A V A L U E I N T E R M E D I A T E B E T W E E N V A L U E S O B T A I N E D W I T H 0.36 A N D 0.86$ L Y S I N E W H E N 1.24$ L Y S I N E WAS F E D . R E S U L T S A L S O C O N F I R M T H E D A T A ON T H R E O N I N E R E T E N T I O N I N T I S S U E P R O T E I N S ( T R I A L 7) W H I C H I N D I C A T E T H A T I N L Y S I N E D E F I C I E N C Y , T H E R E T E N T I O N O F T H E A M I N O A C I D I N T H E L I V E R A N D M U S C L E P R O T E I N I S L E S S T H A N N O R M A L . P R E V I O U S S T U D I E S S H O W E D T H A T E X C R E T I O N O F T H R E O N I N E F R O M T H E R A T WAS I N C R E A S E D M A R K E D L Y B Y F E E D I N G T H E A N I M A L A L Y S I N E - F R E E D I E T ( Y A M A S H I T A A N D S A K U R A I , 1966). R E S U L T S I N D I C A T E T H A T I N D I V I D U A L A M I N O A C I D R E T E N T I O N OR O X I D A T I O N I S A S S O C I A T E D W I T H T H E R A T E O F P R O T E I N S Y N T H E S I S . L I M I T E D P R O T E I N S Y N T H E S I S C A U S E D B Y A L I M I T A T I O N I N D I E T A R Y L Y S I N E R E S U L T S I N A L O S S O F A B S O R B E D T H R E O N I N E W H E R E A S W I T H A D E Q U A T E D I E T A R Y L Y S I N E , P R O T E I N S Y N T H E S I S U T I L I Z E S A L A R G E P R O P O R T I O N O F D I E T A R Y L Y S I N E , T H E R E B Y R E D U C I N G T H E A M O U N T O F T H R E O N I N E A V A I L A B L E F O R O X I D A T I O N . T H R E O N I N E I S KNOWN TO S H A R E T H E R E L A T I V E M E T A B O L I C I N E R T N E S S B E H A V I O U R O F L Y S I N E ( M E I S T E R , 1957), T H E M A J O R C A T A B O L I C P A T H W A Y OF T H R E O N I N E B E I N G T H E F O R M A T I O N O F S U C C I N A T E V I A A L P H A - K E T O G L U T A R I C A C I D ( M E I S T E R , 1965). A L T H O U G H T H E A D D I T I O N O F T H R E O N I N E T O D I E T S C O N T A I N I N G L Y S I N E I N V A R I O U S A M O U N T S D I D N O T I M P R O V E F E E D I N T A K E A N D C H I C K G R O W T H M A R K E D L Y I N T H I S T R I A L ( T A B L E 28), T H E R E T E N T I O N O F T H R E O N I N E I N T H E L I V E R A N D M U S C L E T I S S U E S WAS I N F L U E N C E D B Y T H E L E V E L S O F D I E T A R Y L Y S I N E . HOWEVER, A C H A N G E I N B O D Y W E I G H T I S NOT A R E F L E C T I O N O F T H E C H A N G E I N O N E O R G A N B U T A N A L G E B R A I C S U M O F C H A N G E S I N D I F F E R E N T O R G A N S O F T H E B O D Y . THE O X I D A T I O N A N D R E T E N T I O N S T U D Y WAS F O R A S P E C I F I C P E R I O D O F TWO H O U R S W H E R E A S T H E C H A N G E I N B O D Y W E I G H T WAS M E A S U R E D O V E R A P E R I O D O F 28 D A Y S . THE N U T R I T I O N A L S T A T U S O F T H E A N I M A L S B E F O R E T H E Y WERE P U T ON T H E T E S T D I E T P R O B A B L Y I N F L U E N C E D T H E I R R E S P O N S E TO D I E T A R Y T H R E O N I N E S I N C E S U P P L E M E N T A T I O N O F 0.168$ L - T H R E O N I N E TO A L Y S I N E A D E Q U A T E D I E T F O R F I V E W E E K S WAS S H O W N TO I M P R O V E C H I C K G R O W T H S I G N I F I C A N T L Y I N T R I A L 6. E X C E S S D I E T A R Y L Y S I N E WAS O B S E R V E D T O A F F E C T T H R E O N I N E R E T E N T I O N S L I G H T L Y I N T R I A L 7. THE I M P A C T O F E X C E S S D I E T A R Y L Y S I N E ON T H R E O N I N E R E T E N T I O N WAS M O R E D R A S T I C I N T H I S T R I A L . T H R E O N I N E - U - C R E T E N T I O N I N L I V E R P R O T E I N S WAS L E S S T H A N N O R M A L ( I N T E R M E D I A T E B E T W E E N T H E G R O U P S R E C E I V I N G 0.36 A N D 0.86$ L Y S I N E ) W I T H E X C E S S L Y S I N E . T H R E O N I N E R E T E N T I O N I N M U S C L E P R O T E I N S S H O W E D T H E S A M E P A T T E R N A S I N T H E L I V E R E X C E P T T H A T G R O U P S R E C E I V I N G E X C E S S I V E A N D D E F I C I E N T L Y S I N E D I E T S D I D N O T D I F F E R F R O M E A C H O T H E R M A R K E D L Y I N T H E I R C A P A C I T Y TO R E T A I N T H R E O N I N E . D I F F E R E N C E S I N E X P E R I M E N T A L C O N D I T I O N I N G O F C H I C K S A N D T H E D U R A T I O N O F F E E D I N G T H E E X P E R I M E N T A L D I E T S I N T R I A L S 7 A N D 8 157 P R O B A B L Y A C C O U N T F O R T H E D I S C R E P A N C Y . C H I C K S I N T H E P R E S E N T T R I A L W E R E F E D T H E E X P E R I M E N T A L D I E T S F R O M H A T C H TO 26 D A Y S O F A G E B E F O R E T H E O X I D A T I O N A N D R E T E N T I O N S T U D Y WAS P E R F O R M E D . C H I C K S U S E D I N T R I A L 7 ON T H E O T H E R H A N D R E C E I V E D A C O M M E R C I A L B R O I L E R D I E T F O R 14 D A Y S B E F O R E T H E Y W E R E T R A N S F E R E D TO T H E E X P E R I M E N T A L D I E T S F O R A N O T H E R S I X D A Y S A F T E R W H I C H T H E O X I D A T I O N A N D R E T E N T I O N O F T H R E O N I N E W E R E M E A S U R E D . C H I C K S I N T H E P R E S E N T T R I A L D I D N O T R E C E I V E A N Y F E E D 14 D U R I N G CC^ C O L L E C T I O N W H E R E A S C H I C K S I N T R I A L 7 W E R E R E T U R N E D TO 14, T H E I R F E E D I M M E D I A T E L Y A F T E R I N J E C T I N G T H R E O N I N E - U - C A N D S A C R I F I C E D TWO H O U R S L A T E R . R E S U L T S A G R E E W I T H A N D C O N F I R M T H E O B S E R V A T I O N S I N T R I A L S 6 A N D 14 7 T H A T T H E O X I D A T I O N O F C - L Y S I N E I N C R E A S E D A N D T H E R E T E N T I O N O F T H E L A B E L L E D A M I N O A C I D I N T I S S U E P R O T E I N D E C R E A S E D A S T H E L E V E L O F 14, D I E T A R Y L Y S I N E I N C R E A S E D . T H E R E T E N T I O N O F C - T H R E O N I N E I N T H E C H I C K I S M A R K E D L Y I M P A I R E D B Y L Y S I N E D E F I C I E N C Y A S N O T E D I N T R I A L 7 A N D S L I G H T L Y B Y L Y S I N E T O X I C I T Y . S U M M A R Y A N D C O N C L U S I O N THE E F F E C T S O F D I E T A R Y L Y S I N E C O N T E N T ON T H E M E T A B O L I C T R A N S F O R M A T I O N O F L Y S I N E A N D T H R E O N I N E WERE D E T E R M I N E D . C H I C K S F E D 0.36, 0.86, 1.24$ D I E T A R Y L Y S I N E W E R E I N T R A P E R I T O N E A L L Y I N J E C T E D W I T H L Y S I N E - U - ^ C OR T H R E O N I N E - U - ^ C . R A D I O A C T I V I T Y R E C O V E R Y I N R E S P I R A T O R Y CO^ ,, L I V E R A N D M U S C L E P R O T E I N S W E R E D E T E R M I N E D . R E S U L T S C O N F I R M E D T H E O B S E R V A T I O N S I N T R I A L S 6 A N D 7 T H A T O X I D A T I O N O F L Y S I N E I N C R E A S E D W H I L E L Y S I N E R E T E N T I O N I N L I V E R A N D M U S C L E P R O T E I N S D E C L I N E D W I T H I N C R E A S I N G D I E T A R Y L Y S I N E . T H R E O N I N E O X I D A T I O N WAS I N C R E A S E D A N D 158 R E T E N T I O N D E C R E A S E D B Y L Y S I N E D E F I C I E N C Y A N D T O X I C I T Y , T H E I M P A C T WAS M O R E P R O N O U N C E D I N L Y S I N E D E F I C I E N C Y . GENERAL SUMMARY AND CONCLUSION T H E M E T A B O L I S M O F D E O X Y R I B O N U C L E I C A C I D (DNA), R I B O N U C L E I C A C I D (RNA), P R O T E I N A N D A M I N O A C I D S W E R E S T U D I E D I N C H I C K S F E D D I E T A R Y P R O T E I N S O F D I F F E R E N T N U T R I T I V E V A L U E . B R O I L E R C H I C K S I N A S E R I E S O F T R I A L S WERE F E D D I E T S C O N T A I N I N G W H E A T (15$ D I E T A R Y P R O T E I N ) , W H E A T S U P P L E M E N T E D W I T H G R A D E D L E V E L S O F L Y S I N E , E X C E S S L Y S I N E D I E T S U P P L E M E N T E D W I T H A R G I N I N E , W H E A T - S 0 Y B E A N (15$ D I E T A R Y P R O T E I N ) A N D W H E A T - S O Y B E A N P L U S F I S H M E A L (20$ D I E T A R Y P R O T E I N ) A S S O U R C E S O F D I E T A R Y P R O T E I N . G A S T R O C N E M I U S M U S C L E A N D L I V E R DNA A N D RNA C O N T E N T W E R E M E A S U R E D B Y M E A N S O F U L T R A V I O L E T A B S O R P T I O N T E C H N I Q U E . T I S S U E P R O T E I N S W E R E D E T E R M I N E D B Y S P E C T R O P H O T O M E T R Y , A N D B Y D R Y W E I G H T A F T E R T H E E X T R A C T I O N O F DNA, RNA A N D L . I P I D S F R O M T I S S U E S A M P L E S . I S O T O P E T R A C E R T E C H N I Q U E WAS U S E D TO S T U D Y E F F E C T S O F D I E T A R Y L Y S I N E ON T H E D E G R A D A T I O N A N D S Y N T H E S I S O F T I S S U E P R O T E I N S , RNA S Y N T H E S I S A N D T H E M E T A B O L I C F A T E O F L Y S I N E A N D T H R E O N I N E . R E S U L T S S H O W E D T H A T M U S C L E P R O T E I N / D N A A N D RNA/DNA R A T I O S ( C E L L S I Z E ) I N C R E A S E D M A R K E D L Y W I T H D I E T A R Y L Y S I N E I N T A K E U N T I L T H A T L E V E L (0.86$ O F T H E D I E T ) A T W H I C H B O D Y W E I G H T G A I N A N D G A I N / F E E D W E R E M A X I M A L I N C H I C K S A T 14 A N D 28 D A Y S O F A G E A N D D E C R E A S E D A T D I E T A R Y L Y S I N E L E V E L S H I G H E R T H A N 0.86$. THE R A T I O S I N T H E M U S C L E A L S O I N C R E A S E D B Y I N C R E A S I N G D I E T A R Y P R O T E I N F R O M 15 TO 20$ OR B Y S U P P L E M E N T I N G W H E A T W I T H S O Y B E A N M E A L . 159 L I V E R P R O T E I N / D N A A N D RNA/DNA R A T I O S D E C R E A S E D P R O G R E S S I V E L Y W I T H I N C R E A S E D D I E T A R Y L Y S I N E A N D P R O T E I N C O N T E N T A T 14 D A Y S O F A G E T H E R A T I O S I N C R E A S E D I N L I K E M A N N E R A S T H E M U S C L E A T 28 D A Y S O F A G E L I V E R P R O T E I N / R N A R A T I O A T B O T H 14 A N D 28 D A Y S O F A G E WAS C O N S T A N T A N D I N D E P E N D E N T O F L Y S I N E I N T A K E I N D I C A T I N G T H A T RNA I S C L O S E L Y C O R R E L A T E D W I T H P R O T E I N B I O S Y N T H E S I S . R E S U L T S ON T H E C H I C K M U S C L E A N D L I V E R U N D E R T H E E X P E R I M E N T A L C O N D I T I O N S O F T H I S S T U D Y S U G G E S T T H E F O L L O W I N G C O N C L U S I O N S : (1) T H E Q U A N T I T Y A N D Q U A L I T Y O F D I E T A R Y P R O T E I N S H A V E M A R K E D I M P A C T ON C E L L U L A R OR O R G A N C O N T E N T O F DNA, RNA A N D P R O T E I N I N T H E C H I C K A N D H E N C E T H E S E B I O C H E M I C A L C O N S T I T U E N T S C A N S E R V E NOT O N L Y A S A C R I T E R I O N F O R E V A L U A T I N G D I E T A R Y P R O T E I N S B U T A L S O TO R E V E A L T H E N U T R I T I O N A L S T A T U S O F T H E A N I M A L . (2) M U S C L E G R O W T H I N T H E C H I C K I S B Y A N I N C R E A S E I N C E L L S I Z E A N D NOT I N T H E N U M B E R O F C E L L S OR F I B E R S F R O M H A T C H TO 28 D A Y S O F A G E W H E R E A S T H E L I V E R U N D E R G O E S R A P I D C E L L M U L T I P L I C A T I O N ( H Y P E R P L A S I A ) D U R I N G T H E F I R S T 14 D A Y S O F A G E A N D L I V E R G R O W T H A T 28 D A Y S O F A G E I S M O S T L Y B Y I N C R E A S E I N C E L L S I Z E ( H Y P E R T R O P H Y ) . (3) P R O T E I N A C C U M U L A T I O N WAS D E P R E S S E D M O R E S E V E R E L Y I N B O T H O R G A N S T H A N DNA B Y L Y S I N E A N D P R O T E I N D E F I C I E N C Y . T H I S I N D I C A T E S T H A T , A L T H O U G H C H I C K G R O W T H I S S E V E R E L Y I N H I B I T E D B Y L Y S I N E A N D P R O T E I N D E F I C I E N C Y , M U S C L E A N D L I V E R DNA S Y N T H E S I S C O N T I N U E S A T A F A S T E R R A T E T H A N P R O T E I N S Y N T H E S I S ; I . E . T I S S U E C E L L S I Z E M A Y B E G R E A T L Y R E D U C E D I N C H I C K S U N D E R N O U R I S H E D F R O M O N E D A Y O F A G E B U T NO D E G E N E R A T I O N OR L O S S O F C E L L S OR F I B R E S O C C U R S A N D T H I S R E S U L T P R O B A B L Y E X P L A I N S WHY C H I C K S U N D E R S E V E R E P R O T E I N A N D / O R C A L O R I E D E F I C I E N C Y C A N GROW TO A N O R M A L S I Z E W H E N R E H A B I L I T A T E D ON D I E T S A D E Q U A T E I N P R O T E I N A N D C A L O R I E . E X C E S S D I E T A R Y L Y S I N E D E P R E S S E D C H I C K G R O W T H , F E E D I N T A K E , M U S C L E A N D L I V E R P R O T E I N / D N A A N D RNA/DNA R A T I O S M A R K E D L Y . T H E A D V E R S E E F F E C T S O F E X C E S S L Y S I N E W E R E A L L E V I A T E D B Y S U P P L E M E N T A R Y A R G I N I N E . R E S U L T S D E M O N S T R A T E T H A T W H E N E X C E S S L Y S I N E I S P R E S E N T I N T H E C H I C K D I E T , A R G I N I N E B E C O M E S L I M I T I N G F O R M U S C L E A N D L I V E R C E L L U L A R G R O W T H . D E P R E S S E D T I S S U E ( M U S C L E ) G R O W T H I S E N O U G H TO A C C O U N T F O R D E P R E S S E D C H I C K G R O W T H . H A L F - L I F E V A L U E S O F 7.2, 4.9 A N D 5.2 D A Y S C O R R E S P O N D I N G TO 10.4, 7.1 A N D 7.5 D A Y S T U R N O V E R T I M E , A N D 9.6, 14.0 A N D 13.3$ D A I L Y N E T D E G R A D A T I O N WERE O B S E R V E D F O R L I V E R P R O T E I N S I N C H I C K S (8 TO 15 D A Y S O F A G E ) C O N S U M I N G 0.36, 0.86 A N D 1.24$ L Y S I N E R E S P E C T I V E L Y . L I V E R P R O T E I N R A T E O F S Y N T H E S I S WAS 20.0, 49.2 A N D 52.9 M G . / D A Y R E S P E C T I V E L Y . T H E T U R N O V E R R A T E S O F B O T H T H E M U S C L E A N D L I V E R P R O T E I N S I N C R E A S E D I N A M A N N E R S I M I L A R TO T H E G A I N I N B O D Y W E I G H T W I T H T H E L E V E L O F D I E T A R Y L Y S I N E . R E S U L T S S U G G E S T T H A T T H E R A T E O F P R O T E I N S Y N T H E S I S A N D D E G R A D A T I O N I N T H E T I S S U E O F L Y S I N E D E F I C I E N T C H I C K I S C O N S I D E R A B L Y L O W E R T H A N I N T H E L I V E R O F C H I C K S F E D A D E Q U A T E L Y S I N E . L Y S I N E D E F I C I E N C Y A L S O D E P R E S S E D RNA S Y N T H E S I S I N T H E L I V E R M A R K E D L Y A N D I N T H E M U S C L E S L I G H T L Y I N C H I C K S A T E I G H T D A Y S O F A G E . O X I D A T I O N O F L Y S I N E I N C R E A S E D W H I L E L Y S I N E R E T E N T I O N I N L I V E R A N D M U S C L E P R O T E I N S D E C L I N E D W I T H I N C R E A S I N G D I E T A R Y L Y S I N E . T H R E O N I N E O X I D A T I O N WAS I N C R E A S E D A N D R E T E N T I O N D E C R E A S E D B Y L Y S I N E D E F I C I E N C Y A N D T O X I C I T Y , T H E I M P A C T WAS G R E A T E R I N L Y S I N E D E F I C I E N C Y . R E S U L T S S U G G E S T T H A T WHEN L Y S I N E I S P R E S E N T I N T H E C H I C K D I E T I N S U B O P T I M A L A M O U N T S , A L A R G E P R O P O R -T I O N O F T H I S A M I N O A C I D I S U S E D F O R P R O T E I N S Y N T H E S I S , I N C R E A S E D O X I D A T I O N OF T H E C A R B O N S K E L E T O N TO C A R B O N D I O X I D E O C C U R S A S T H E D I E T A R Y S U P P L Y E X C E E D S T H E A N I M A L N E E D S F O R P R O T E I N S Y N T H E S I S . L I M I T E D P R O T E I N S Y N T H E S I S C A U S E D B Y A L I M I T A T I O N I N D I E T A R Y L Y S I N E R E S U L T S I N A L O S S O F T H R E O N I N E W H E R E A S W I T H A D E Q U A T E D I E T A R Y L Y S I N E , P R O T E I N S Y N T H E S I S U T I L I Z E S A L A R G E P R O P O R T I O N O F D I E T A R Y L Y S I N E , T H E R E B Y R E D U C I N G T H E A M O U N T S O F T H R E O N I N E A V A I L A B L E F O R O X I D A T I O N . BIBLIOGRAPHY A D D I S , T., L. J. Poo A N D W. LEW, 1936. THE Q U A N T I T Y O F P R O T E I N L O S T B Y T H E V A R I O U S O R G A N S A N D T I S S U E S O F T H E B O D Y D U R I N G A F A S T . J . B I O L . CHEM. 115:111-116. A D D I S , T., L. J. Poo A N D W. LEW, 1936. P R O T E I N L O S S F R O M L I V E R D U R I N G A TWO D A Y F A S T . J . B I O L . CHEM. 115:117-118. A D D I S , T., L. J. Poo A N D W. LEW, 1936. 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FATE O F T H R E O N I N E A N D L E U C I N E I N R A T S F E D T H R E O N I N E - D E F I C I E N T D I E T S . J . NUTR. 97:367-374. Y A M A S H I T A , K. A N D K. A S H I D A , 1969. L Y S I N E M E T A B O L I S M I N R A T S F E D L Y S I N E - F R E E D I E T . J. NUTR. 99:267-273. Y A M A S H I T A , K. A N D K. A S H I D A , 1971. E F F E C T O F E X C E S S I V E L E V E L S O F L Y S I N E A N D T H R E O N I N E ON T H E M E T A B O L I S M O F T H E S E A M I N O A C I D S I N R A T S . J. NUTR. 101:1607-1614. Y A M A S H I T A , M. A N D Y. S U K U R A I , 1966. S T U D I E S ON N U T R I T I O N O F T H R E O N I N E 3. R E L A T I O N S H I P B E T W E E N T H E A B I L I T Y O F R A T L I V E R F O R D E C O M P O S I N G T H R E O N I N E A N D T H E E X C R E T I O N O F T H R E O N I N E I N U R I N E , E S P E C I A L L Y , T H E E F F E C T O F L Y S I N E C O N T E N T I N T H E D I E T ON I T . J. J A P . SOC. FOOD NUTR. 9:218-221. YOUNG, V. R. A N D S. D. A L E X I S , 1968. I N V I T R O A C T I V I T Y O F R I B O S O M E S A N D RNA C O N T E N T O F S K E L E T A L M U S C L E I N Y O U N G R A T S F E D A D E Q U A T E OR LOW P R O T E I N . J. NUTR. 96:255-262. YOUNG, V. R., S. C. S T O T H E R S A N D G. V I L A I R E , 1971. S Y N T H E S I S A N D D E G R A D A T I O N O F M I X E D P R O T E I N S , A N D C O M P O S I T I O N C H A N G E S I N S K E L E T A L M U S C L E O F M A L N O U R I S H E D A N D R E F E D R A T S . J . NUTR. 101 :1379-1390. Y O S H I D A , A., P. M. B. L E U N G , Q. R. ROGERS A N D A. E. H A R P E R , 1966. E F F E C T O F A M I N O A C I D I M B A L A N C E ON T H E F A T E O F T H E L I M I T I N G A M I N O A C I D . J . NUTR. 89:80-90. Y U I L E , C. L., F. V. L U C A S , J . P. OLSON A N D A. B. S H A P I R O , 1959. P L A S M A P R O T E I N T U R N O V E R A N D T I S S U E E X C H A N G E . I N F L U E N C E O F D I E T A R Y P R O T E I N A N D P R O T E I N D E P L E T I O N . J . E X P . M E D . 109:173-186. Z I M M E R M A N , R. A. A N D H. M. S C O T T , 1965. I N T E R R E L A T I O N S H I P O F P L A S M A A M I N O A C I D L E V E L S A N D W E I G H T G A I N I N T H E C H I C K A S I N F L U E N C E D B Y S U B - O P T I M A L A N D S U P E R - O P T I M A L D I E T A R Y C O N C E N T R A T I O N O F S I N G L E A M I N O A C I D S . J . N U T R . 87:13-18. APPENDIX APPENDIX TABLE I . - - A N A L Y S I S O F V A R I A N C E O F A V E R A G E B O D Y W E I G H T G A I N , F E E D I N T A K E , G A S T R O C N E M I U S M U S C L E W E I G H T , L I V E R W E I G H T A N D L I V E R W E I G H T P E R 100 G. B O D Y W E I G H T O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R TWO W E E K S ( T R I A L l ) S O U R C E O F MEAN S Q U A R E VAR1 A T 1 ON D F BODY WT. G A I N F E E D I N T A K E M U S C L E WT. L I V E R WT. L I V E R WT./100G. BODY WT. T R E A T M E N T 4 2106.76** 6087.70** 354630.84 4.10** 0.93** REPL1 C A T 1 ON 2 63.65 364.63 89095.45 0.56 0.07 E R R O R 8 52.87 151.34 51401.94 0.28 0.05 T O T A L 14 * S T A T i S T i C A L L Y S I G N I F I C A N T A T P -=0.05. * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P "=0.01. APPENDIX TABLE I I . - - A N A L Y S I S O F V A R I A N C E O F DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N I N O N E G R A M O F G A S T R O C N E M I U S M U S C L E O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R TWO W E E K S ( T R I A L l ) S O U R C E O F V A R I A T I O N D F MEAN S Q U A R E DNA RNA P R O T E I N T R E A T M E N T 4 0.0015 0.07 683.25 R E P L 1 C A T 1 ON 2 0.0008 110.19 E R R O R 8 0.0047 0.02 300.96 T O T A L 14 184 APPENDIX TABLE I I I . - - A N A L Y S I S O F V A R I A N C E O F T O T A L DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N I N T H E G A S T R O C N E M I U S M U S C L E O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R TWO W E E K S ( T R I A L l ) S O U R C E O F V A R I A T I O N D F MEAN S Q U A R E DNA RNA P R O T E 1 N T R E A T M E N T 4 0.1035** 1 .2731** 8649.24** R E P L I C A T I ON 2 0.0368 0.3049 1348.30 E R R O R 8 0.0135 0.1469 877.70 T O T A L 14 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P<=0.01. APPENDIX TABLE I V . - - A N A L Y S I S O F V A R I A N C E O F DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N I N O N E G R A M O F L I V E R O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R TWO W E E K S ( T R I A L l ) S O U R C E O F MEAN S Q U A R E V A R I A T I O N D F DNA RNA P R O T E 1 N T R E A T M E N T 4 0.49** 2.22** 189.42 REPL1 C A T 1 ON 2 0.03 0.08 1 02.85 E R R O R 8 0.03 0.27 168.28 T O T A L 14 * * S T A T I S T I C A L L Y S I GN1 F1 C A N T A T P -=0.01 . APPENDIX TABLE V . — A N A L Y S I S O F V A R I A N C E O F T O T A L DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N I N T H E L I V E R S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R TWO W E E K S ( T R I A L I) S O U R C E O F MEAN S Q U A R E V A R I A T 1 ON D F DNA RNA P R O T E I N T R E A T M E N T 4 36.94** 372.01** 113924.41** REPL1 C A T 1 ON 2 5.26 50.17 11672.15 E R R O R 8 1 .76 16.49 4230.50 T O T A L 14 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.01. APPENDIX TABLE V I . - - A N A L Y S I S O F V A R I A N C E O F T O T A L L I V E R DNA, RNA A N D P R O T E I N C O N T E N T P E R 100 G. B O D Y W E I G H T O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R TWO W E E K S ( T R I A L l ) S O U R C E O F VARI A T I ON D F MEAN S Q U A R E DNA RNA P R O T E 1 N T R E A T M E N T 4 16.41** 110.42** 31421.17** REPL1 C A T 1 ON 2 0.26 5.93 62.03 E R R O R 8 0.63 7.78 3250.95 T O T A L 14 * * S T A T I S T I C A L L Y S I G N I F I C A N T AT:P <0.01. APPENDIX TABLE VI I . - - A N A L Y S I S O F V A R I A N C E O F A V E R A G E B O D Y W E I G H T G A I N , F E E D I N T A K E , G A S T R O C N E M I U S M U S C L E W E I G H T , L I V E R W E I G H T A N D L I V E R W E I G H T P E R 100 G. B O D Y W E I G H T O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R F O U R W E E K S ( T R I A L l ) S O U R C E O F MEAN S Q U A R E VARI A T I ON r !-DF BODY WT. G A I N F E E D I N T A K E M U S C L E WT. L I V E R WT. L I V E R WT./100G. BODY WT. T R E A T M E N T 3(3)1 6598.31** 32567.44** 7178347.68** 47.53** 0.23 REPL1 C A T 1 ON 2(2)1 130.00 931.46 94843.25 1 .34 --AGE 3 3551.35 40039.45 80223.03 2.33 0.08 ERROR 39(6)1 128.23 925.47 — T O T A L 47(11 )1 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01. 1 D E G R E E O F F R E E D O M F O R M U S C L E W E I G H T , L I V E R W E I G H T A N D L I V E R W E I G H T P E R 100 G. B O D Y W E I G H T I S I N P A R E N T H E S E S . APPENDIX TABLE V I I I . — ANALYSIS OF VARIANCE OF DNA, RNA AND PROTEIN CONCENTRATION IN ONE GRAM OF GASTROCNEMIUS MUSCLE OF CHICKS FED GRADED LEVELS OF LYSINE FOR FOUR WEEKS (TRIAL l ) SOURCE OF VAR1 ATI ON DF MEAN SQUARE DNA RNA PROTEIN TREATMENT 3 0.0187 0.866 1652.42** REPLlCAT 1 ON 2 0.0161 0.020 151.90 ERROR 6 0.0131 0.028 96.35 TOTAL 11 STATISTICALLY SIGNIFICANT AT;:P •*= 0.01. 190 APPENDIX TABLE I X . - - A N A L Y S I S O F V A R I A N C E O F T O T A L DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N I N T H E G A S T R O C N E M I U S M U S C L E O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R F O U R W E E K S ( T R I A L l ) S O U R C E O F V A R I A T l O N D F MEAN S Q U A R E DNA RNA P R O T E I N T R E A T M E N T 3 2.12** 12.68** 288233.89** R E P L I C A T 1 ON 2 0.19 0.37 10872.77 E R R O R 6 0.15 0.19 4933.18 T O T A L 11 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P "=0.01. APPENDIX TABLE X . - - A N A L Y S I S O F V A R I A N C E O F DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N I N O N E G R A M O F L I V E R O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R F O U R W E E K S ( T R I A L l ) S O U R C E O F MEAN S Q U A R E VAR1 A T I ON D F DNA RNA P R O T E I N T R E A T M E N T 3 0.066 0.56 28.16 R E P L I C A T I O N 2 0.010 0.25 62.85 E R R O R 6 0.130 0.71 393.70 T O T A L 11 APPENDIX TABLE X I . - - A N A L Y S I S O F V A R I A N C E O F T O T A L DNA, RNA A N D P R O T E I N C O N C E N T R A T I O N I N T H E L I V E R S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R F O U R W E E K S ( T R I A L I) S O U R C E O F VAR1 A T 1 ON D F MEAN S Q U A R E DNA RNA P R O T E I N T R E A T M E N T 3 211.50** 2436.72** 997634.26** REPL1 C A T 1 ON 2 13.88 121.83 63414.29 E R R O R 6 15.73 178.01 72352.69 T O T A L 11 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.01. APPENDIX TABLE XI I . - - A N A L Y S I s O F V A R I A N C E O F T O T A L L I V E R DNA, RNA A N D P R O T E I N C O N T E N T P E R TOO G. B O D Y W E I G H T O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E F O R F O U R W E E K S ( T R I A L l ) S O U R C E O F VAR1 A T 1 ON D F MEAN S Q U A R E DNA RNA P R O T E I N T R E A T M E N T 3 0.17 24.59 5908.46 REPL1 C A T 1 ON 2 0.15 3.22 1124.7-0 E R R O R 6 1.19 11 .24 4350.29 T O T A L 11 APPENDIX TABLE XI I I . - - A N A L Y S I S O F V A R I A N C E O F G A S T R O C N E M I U S M U S C L E W E I G H T A N D T O T A L M U S C L E DNA, RNA A N D P R O T E I N C O N T E N T I N C H I C K S F E D A D E Q U A T E A N D E X C E S S L Y S I N E F R O M O N E D A Y TO 14 D A Y S O F A G E ( T R I A L 2) S O U R C E O F MEAN S Q U A R E VAR1 A T 1 ON D F M U S C L E WT. DNA RNA P R O T E 1 N T R E A T M E N T 2 378990.49* 0.125 1.845* 14565.80** REPL1 C A T 1 ON 2 16106.07 0.005 0.065 142.40 E R R O R 4 28579.56 0.020 0.135 203.18 T O T A L 8 *STAT 1 S T 1 C A L L Y S 1 GN1F1 C A N T A T P < 0. 05. * * S T A T I S T I C A L L Y S 1 G N 1 F1 C A N T A T ? < 0. 01 . APPENDIX TABLE X I V . - - A N A L Y S I S O F V A R I A N C E O F L I V E R W E I G H T A N D T O T A L L I V E R DNA, RNA A N D P R O T E I N C O N T E N T I N C H I C K S F E D A D E Q U A T E A N D E X C E S S L Y S I N E F R O M O N E D A Y TO 14 D A Y S O F A G E ( T R I A L 2) S O U R C E O F MEAN S Q U A R E VAR1 A T 1 ON O F L I V E R WT. DNA RNA P R O T E I N T R E A T M E N T 2 3.32* 25.28** 449.75** 88809.17** R E P L 1 C A T I O N 2 0.12 2.99 23.77 7833.99 E R R O R 4 0.44 2.18 26.19 5074.06 T O T A L 8 * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.05. * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01. APPENDIX TABLE X V . - - A N A L Y S I S O F V A R I A N C E O F G A S T R O C N E M I U S M U S C L E W E I G H T A N D T O T A L M U S C L E DNA, RNA A N D P R O T E I N C O N T E N T I N C H I C K S F E D A D E Q U A T E L Y S I N E A N D E X C E S S L Y S I N E W I T H A R G I N I N E F R O M 14 TO 28 D A Y S O F A G E ( T R I A L 2) S O U R C E OF MEAN S Q U A R E V A R I A T I O N D F M U S C L E WT. DNA RNA P R O T E I N T R E A T M E N T 2 2487440.30 0.16 1 .47 115001 .15 REPL1 C A T 1 ON 2 103484.57 0.14 0.04 2055.75 E R R O R 4 470623.62 0.25 1 .28 44035.27 T O T A L 8 APPENDIX TABLE X V I . - - A N A L Y S I S O F V A R I A N C E O F L I V E R W E I G H T L I V E R DNA, RNA A N D P R O T E I N C O N T E N T I N C H I C K S F E D A D E Q U A T E A N D E X C E S S L Y S I N E W I T H A R G I N I N E F R O M 14 TO 28 D A Y S O F A G E A N D T O T A L L YS I NE ( T R I A L 2) S O U R C E O F . MEAN S Q U A R E VAR 1 A T I ON D F L I V E R WT. DNA RNA P R O T E I N T R E A T M E N T 2 14.16** 89.84* 361.08 171690.96 R E P L I C A T 1 ON 2 0.27 15.06 1 62.07 164825.78 E R R O R 4 1.15 12.45 140.22 36934.66 T O T A L 8 * S T A T 1 S T 1 C A L L Y S 1GN 1 F1 C A N T A T P < 0.05. * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P «=0.01. APPENDIX TABLE XV I I . - - A N A L Y S I S O F V A R I A N C E O F A V E R A G E B O D Y W E I G H T G A I N A N D F E E D I N T A K E O F C H I C K S F E D D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y A N D Q U A L I T Y F O R TWO A N D F O U R W E E K S ( T R I A L 3) MEAN S Q U A R E 2 WK. 4 WK. S O U R C E O F BODY WT. F E E D BODY WT. F E E D VAR1 A T 1 ON D F GA 1 N 1NTAKE GA I N I N T A K E T R E A T M E N T 3 7736.43** 13794.39** 93281.22** 246161.97** R E P L 1 C A T I O N 2 208.08 460.85 142.89 3314.71 ERROR 6 48.93 131.37 405.05 791.35 T O T A L 11 S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01 . APPENDIX TABLE XV I I I . - - A N A L Y S I s O F V A R I A N C E O F G A S T R O C N E M I U S M U S C L E W E I G H T A N D T O T A L M U S C L E DNA, RNA A N D P R O T E I N I N C H I C K S F E D D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y A N D Q U A L I T Y F O R TWO W E E K S ( T R I A L 3). S O U R C E O F MEAN S Q U A R E VAR1 A T 1 ON D F M U S C L E WT. DNA RNA P R O T E I N T R E A T M E N T .3 605191.48** 0.16** 2.96** 19097.26** REPL1 C A T 1 ON 2 38423.58 0.015 0.05 600.80 E R R O R 6 9910.99 0.012 0.13 269.91 T O T A L 11 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.01. APPENDIX TABLE X I X . - - A N A L Y S I S O F V A R I A N C E O F L I V E R W E I G H T A N D T O T A L L I V E R DNA, RNA A N D P R O T E I N I N C H I C K S F E O D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y A N D Q U A L I T Y F O R TWO W E E K S ( T R I A L 3) S O U R C E O F MEAN S Q U A R E V A R I A T I O N D F L I V E R WT. DNA RNA P R O T E I N T R E A T M E N T 3 6.72** 53.01** 511.39** 178897.19** R E P L I C A T I O N 2 0.52 6.01 64.10 8454.61 E R R O R 6 0.12 2.03 16.21 3256.12 T O T A L 11 S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01. APPENDIX TABLE X X . - - A N A L Y S I S O F V A R I A N C E O F G A S T R O C N E M I U S M U S C L E W E I G H T A N D T O T A L M U S C L E DNA, RNA A N D P R O T E I N I N C H I C K S F E D D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y A N D Q U A L I T Y F O R F O U R W E E K S ( T R I A L 3) S O U R C E O F MEAN S Q U A R E V A R 1 A T I ON D F M U S C L E WT. DNA' RNA P R O T E I N T R E A T M E N T 3 21606395.61** 3.54** 52.73* 1192220.79** R E P L 1 C A T 1 ON 2 305528.06 0.10 0.155 11644.02 E R R O R 6 65053.69 0.15 0.575 3319.98 T O T A L 11 * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.05. * * S T A T I S T I C A L ' L Y S I G N I F I C A N T A T P < 0.01 . APPENDIX TABLE XXI. — A N A L Y S I S O F V A R I A N C E O F L I V E R W E I G H T A N D T O T A L L I V E R DNA, RNA A N D P R O T E I N I N C H I C K S F E D D I E T A R Y P R O T E I N S D I F F E R E N T I N Q U A N T I T Y A N D Q U A L I T Y F O R F O U R W E E K S ( T R I A L 3) S O U R C E O F MEAN S Q U A R E VAR1 A T 1 ON D F L I V E R WT. DNA RNA P R O T E 1 N T R E A T M E N T 3 83.58** 348.26** 3871.31** 1992883.75** REPL1 C A T 1 ON 2 3.42 51 .86 199.09 191146.40 E R R O R 6 2.06 24.91 117.32 69800.05 T O T A L 11 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01. 203 APPENDIX TABLE XXI I . - - A N A L Y S I S O F V A R I A N C E O F P R O T E I N I N O N E G R A M O F M I X E D T I B I A M U S C L E S , R A D I O A C T I V I T Y O F M U S C L E P R O T E I N S ( D P M / G . T I S S U E ) A N D S P E C I F I C A C T I V I T Y O F M U S C L E P R O T E I N S ( D P M / M G . . P R O T E I N ) F R O M C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - C ( T R I A L 4) S O U R C E O F MEAN S Q U A R E VAR1 AT 1 O N O F P R O T E I N R A D I O A C T i v I T Y S P E C I F I C A C T I v i T Y T R E A T M E N T 2 498.53** 7489765.01** 1000. 57** P E R I O D 3 152.50** 27568720.98** 1 824. 48** TRT. X P E R I O D (6)1 5905350.23** 239. 78** E R R O R 30(24)1 12.11 96073.92 12. 75 T O T A L 35 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <=0.01. D E G R E E O F F R E E D O M L O C A T E D I N P A R E N T H E S E S I S F O R R A D I O A C T I V I T Y A N D S P E C I F I C A C T I V I T Y . APPENDIX TABLE XXI I I . - - A N A L Y S I S O F V A R I A N C E O F RNA I N O N E G R A M O F M I X E D T I B I A M U S C L E S , R A D I O A C T I V I T Y R E C O V E R Y ( D P M / G . P R O T E I N / P E R C E N T D O S E ) A N D R A D I O A C T I V 1 T Y / M G . RNA O F T H E M U S C L E S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - ^ 4 C ( T R I A L 4) MEAN S Q U A R E S O U R C E O F V A R I A T I O N D F RNA RAD 1 OACT 1V 1TY R E C O V E R Y R A D I O A C T I V I T Y / M G . RNA T R E A T M E N T 2 0.61* 0.51** 3180125.32** P E R I O D 3 0.53* REPL1 C A T 1 ON (2)1 0.03 221746.43 E R R O R 30(28)1 0.12 0.005 93301 .13 T O T A L 35 * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.05. * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.01. 1 D E G R E E O F F R E E D O M L O C A T E D I N P A R E N T H E S E S I S F O R R A D I O A C T I V I T Y R E C O V E R Y A N D R AD I O A C T I V I T Y / M G . RNA. APPENDIX TABLE XXIV . - - A N A L Y S i s O F V A R I A N C E O F P R O T E I N I N O N E G R A M O F L I V E R , R A D I O A C T I V I T Y O F L I V E R P R O T E I N S ( D P M / G . T I S S U E ) A N D S P E C I F I C A C T I V I T Y O F L I V E R P R O T E I N S ( D P M / M G . P R O T E I N ) F R O M C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - ^ C ( T R I A L 4) S O U R C E O F MEAN S Q U A R E VARI A T 1 ON D F P R O T E 1 N R A D 1 O A C T I V 1TY S P E C i F i c A C T I V I T Y T R E A T M E N T 2 443.36** 149082662.27** 6208.12** P E R I O D 3 2372.20** 203622391 .26** 23772.94** TRT. X P E R I O D (6)1 13517987.40** 612.30** E R R O R 30(24)1 68.58 496827.13 90.91 T O T A L 35 * * S T A T 1 S T 1 C A L L Y S I G N I F I C A N T A T P < 0.01 . D E G R E E O F F R E E D O M L O C A T E D I N P A R E N T H E S E S I S F O R R A D I O A C T I V I T Y A N D S P E C I F I C A C T I V I T Y . APPENDIX TABLE X X V . — A N A L Y S I S O F V A R I A N C E O F T O T A L L I V E R P R O T E I N A N D RNA F R O M C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - ^ 4 C ( T R I A L 4) S O U R C E O F V A R I A T I O N O F MEAN S Q U A R E P R O T E I N RNA T R E A T M E N T 2 692254.82** 3199.32** P E R I O D 3 87374.42 265.70 E R R O R 30 33195.29 133.71 T O T A L 35 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.01. APPENDIX TABLE X X V I . - - A N A L Y S i s O F V A R I A N C E O F RNA I N O N E G R A M O F L I V E R , R A D I O A C T I V I T Y R E C O V E R Y ( D P M / G . P R O T E I N / P E R C E N T D O S E ) A N D R A D I O-A C T I V I T Y / M G . RNA O F T H E L I V E R S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - C ( T R I A L 4) MEAN S Q U A R E S O U R C E O F VAR 1 A T 1 ON D F RNA R A D I O A C T I v i T Y R E C O V E R Y R A D i O A C T i v i T Y / M G . RNA T R E A T M E N T 2 1 .88* 3.15** 8404073967.77** P E R I O D 3 5.08** REPL1 C A T 1 ON (2)1 0.13 1126748.74 ERROR 30(28)1 0.38 0.04 43645500.25 T O T A L 35 * S T A T I S T l C A L L Y S I G N I F I C A N T A T P < 0.05. * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01. 1 _ D E G R E E O F F R E E D O M L O C A T E D I N P A R E N T H E S E S I S F O R R A D I O A C T I V I T Y R E C O V E R Y A N D RAD I O A C T I V I T Y / M G . RNA. APPENDIX TABLE XXVI I . - - A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y I N M U S C L E H O M O G E N A T E ( D P M / G . T I S S U E ) , RNA E X T R A C T ( D P M / G . T I S S U E ) , RNA E X T R A C T E X P R E S S E D A S A P E R C E N T A G E O F R A D I O A C T I V I T Y I N M U S C L E H O M O G E N A T E , A N D S P E C I F I C A C T I V I T Y ( D P M / M G . RNA) O F C H I C K S F E D G R A D E D L E V E L S OF L Y S I N E A N D I N J E C T E D W I T H U R A C I L - U - ^ 4 C ( T R I A L 5) S O U R C E O F MEAN S Q U A R E V A R I A T I O N O F H O M O G E N A T E RNA E X T R A C T RNA E X T R A C T / H O M O G E N A T E S P E C 1 F I C A C T i v i T Y T R E A T M E N T 2 26172.27* 443.92 1.17 165 .12 1 ND1V1 D U A L 2 3105.02 2502.61* 5.60 743 .67 ERROR 4 3384.92 352.74 2.53 145 .29 T O T A L 8 * S T A T I S T I C A L L Y S I G N I F I C A N T A T P -=0.05. no o 00 APPENDIX TABLE XXV I I I . - - A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y I N L I V E R H O M O G E N A T E ( D P M / G . T I S S U E ) , RNA E X T R A C T ( D P M / G . T I S S U E ) , RNA E X T R A C T E X P R E S S E D A S A P E R C E N T A G E O F R A D I O A C T I V I T Y I N L I V E R H O M O G E N A T E , A N D S P E C I F I C A C T I V I T Y ( D P M / M G . RNA) O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H U R A C I L - U - ^ C ( T R I A L 5) S O U R C E O F MEAN S Q U A R E V A R I A T I O N D F H O M O G E N A T E RNA E X T R A C T RNA E X T R A C T / H O M O G E N A T E S P E C I F I C A C T I V I T Y T R E A T M E N T 2 37276.81 15283.78** 19.66* 376.92** I N D I V I D U A L 2 2365.82 749.68 1.31 5.00 E R R O R 4 11350.65 325.40 2.25 11.06 T O T A L 8 * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.05. * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.01. APPENDIX TABLE X X I X . - - A N A L Y S I S O F V A R I A N C E O F A V E R A G E B O D Y W E I G H T G A I N , F E E D I N T A K E , A N D R A D I O A C T I V I T Y R E C O V E R Y I N CO2 F R O M C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D W I T H T H R E O N I N E A N D A R G I N I N E ( T R I A L 6) MEAN S Q U A R E S O U R C E O F V A R I A T I O N D F BODY WT. G A I N F E E D I N T A K E RAD I O A C T I V I T Y R E C O V E R Y T R E A T M E N T 6 49370.24** 21 9826.60** 0.2071** R E P L I C A T 1 ON 2 2295.70 5458.71 E R R O R 12 2777.02 17874.71 0.0165 T O T A L 20 * * S T A T I S T I C A L L Y S I G N I F I C A N T AT :P < 0 . 0 1 . APPENDIX TABLE X X X . - - A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y R E C O V E R Y I N A C I D P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N T H E T I B I A M U S C L E S . O F L Y S I N E A N ( ( T R I A L 7) 1 4. C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - C MEAN S Q U A R E S O U R C E O F VARI A T 1 ON O F Ac I D I N S O L U B L E Ac 1 D S O L U B L E A C I D AC I D I N S O L U B L E S O L U B L E T R E A T M E N T 2 0.0888 0.0402* 89.63 REPL1 C A T 1 ON 1 0.0004 0.0001 0.74 E R R O R 2 0.0368 0.00145 10.09 T O T A L 5 * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.05. APPENDIX TABLE X X X I . — A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y R E C O V E R Y I N A C I D I N S O L U B L E P R O T E I N S , A C I D S O L U B L E P R O T E I N S , A N D A C I D I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N T H E L I V E R S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - L ) - ^4C ( T R I A L 7) MEAN S Q U A R E S O U R C E O F VAR1 A T 1 ON D F Ac 1 D 1 N S O L U B L E Ac 1 D S O L U B L E A C I D AC I D I N S O L U B L E S O L U B L E T R E A T M E N T 2 0.6997 0.00465* 46.58* REPL1 C A T 1 ON 1 0.2481 0.00020 0.69 E R R O R 2 0.0797 0.00005 0.99 T O T A L 5 * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.05. APPENDIX TABLE XXXI I . - - A N A L Y S i s O F V A R I A N C E O F R A D I O A C T I V I T Y R E C O V E R Y I N A C I D I N S O L U B L E P R O T E I N S , A C I D S O L U B L E P R O T E I N S , A N D A C I D I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N T H E T I B I A M U S C L E S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H T H R E O N I N E - U — ^ 4C . ( T R I A L 7) MEAN S Q U A R E S O U R C E O F VAR1 A T I ON D F Ac 1 D 1 N S OL U B L E Ac 1 D S O L U B L E A C I D A C I D I N S O L U B L E S O L U B L E T R E A T M E N T 2 0.245* 0.00515** 265.79** REPL1 C A T 1 ON 3 0.0075 0.00013 8.43 E R R O R 6 0.028 0.00013 15.36 T O T A L 11 * S T A T I S T I C A L L Y S I G N I F I C A N T A T : P -=0.05. * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P ""=0.01. APPENDIX TABLE XXXI I I . — A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y R E C O V E R Y I N A C I D I N S O L U B L E P R O T E I N S , A C I D S O L U B L E P R O T E I N S , A N D A C I D I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N T H E L I V E R S O F C H I C K S F E D G R A D E D . L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H T H R E ON INE-U- C ( T R I A L 7) MEAN S Q U A R E S O U R C E O F VAR I.AT 1 ON D F Ac I D I N S O L U B L E Ac 1 D S O L U B L E A C I D A C I D I N S O L U B L E S O L U B L E T R E A T M E N T 2 0.2650** 0.0253** 322.07** REPL1 C A T 1 ON 3 0.013 0.0008 3.66 E R R O R 6 0.008 0.00035 2.01 T O T A L 11 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01. APPENDIX TABLE X X X I V . - - A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y R E C O V E R Y I N R E S P I R A T O R Y CO;?., A C I D I N S O L U B L E P R O T E I N S , A C I D S O L U B L E P R O T E I N S , A N D A C I D I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N T H E T I B I A M U S C L E S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - U C ( T R I A L 8) MEAN S Q U A R E  S O U R C E O F A C I D A C I D A C I D A C I D V A R I A T I O N D F COg I N S O L U B L E S O L U B L E I N S O L U B L E S O L U B L E T R E A T M E N T 2 2.40* 0.315** 0.3745** 83.57** R E P L I C A T I O N 2 0.51 0.0025 0.05 E R R O R 4 0.26 0.0037 0.0003 0.09 T O T A L 8 * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.05. * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P <0.01. APPENDIX TABLE X X X V . - - A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y R E C O V E R Y I N A C I D I N S O L U B L E P R O T E I N S , A C I D S O L U B L E P R O T E I N S A N D A C I D I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N T H E L I V E R S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H L Y S I N E - U - ^4C ( T R I A L 8) MEAN S Q U A R E A C I D A C I D A C I D S O L U B L E I N S O L U B L E S O L U B L E 0.1065** 0.0003 0.0015 158.46** 1 .06 2.00 S O U R C E O F V A R I A T I O N D F Ac I D I N S O L U B L E T R E A T M E N T 2 10.545** R E P L I C A T I O N 2 0.005 E R R O R 4 0.015 T O T A L 8 * * S T A T 1 S T I C A L L Y S I G N I F I C A N T A T ; P < 0.01. APPENDIX TABLE XXXV I . - - A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y R E C O V E R Y I N R E S P I R A T O R Y COg, A C I D I N S O L U B L E P R O T E I N S , A C I D S O L U B L E P R O T E I N S , A N D A C I D I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N T H E T I B I A M U S C L E S O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H T H R E O N I NE-U-^C ( T R I A L 8) MEAN S Q U A R E  S O U R C E O F A C I D A C I D A C I D A C I D V A R I A T I O N D F CO2 I N S O L U B L E S O L U B L E I N S O L U B L E S O L U B L E T R E A T M E N T 2 0.4832 2.4150** 0.06905** 159.86** R E P L I C A T I O N 2 0.0734 0.00035 0.00135 4.02 E R R O R 4 0.0849 0.0007 0.002775 3.86 T O T A L 8 **S T A T I S T I C A L L Y S I G N I F I C A N T A T P <S 0.01 . APPENDIX TABLE XXXVI I . - - A N A L Y S I S O F V A R I A N C E O F R A D I O A C T I V I T Y R E C O V E R Y I N A C I D I N S O L U B L E P R O T E I N S A N D A C I D S O L U B L E P R O T E I N S , A N D A C I D I N S O L U B L E P R O T E I N S R E L A T I V E TO A C I D S O L U B L E P R O T E I N S I N T H E L I V E R O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E A N D I N J E C T E D W I T H T H R E O N I N E - U " ^ 4 C ( T R I A L 8) MEAN S Q U A R E  S O U R C E O F A C I D A C I D A C I D A C I D V A R I A T I O N D F I N S O L U B L E S O L U B L E I N S O L U B L E S O L U B L E T R E A T M E N T 2 28.9614** 0.0792** 738.3316** R E P L I C A T I O N 2 0.0027. 0.0009 8.9224 E R R O R 4 0.0116 0.00015 3.2340 T O T A L 8 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01. APPENDIX TABLE XXXV I I I . - - A N A L Y S I S O F V A R I A N C E O F A V E R A G E B O D Y W E I G H T G A I N A N D F E E D I N T A K E O F C H I C K S F E D G R A D E D L E V E L S O F L Y S I N E W I T H T H R E O N I N E ( T R I A L 8) S O U R C E O F MEAN S Q U A R E VARI A T I ON D F BODY WT. G A I N F E E D I N T A K E T R E A T M E N T 9 45687.55** 232933.95** REPL1 C A T 1 ON 2 173.63 4198.90 E R R O R 1 8 914.22 5752.38 T O T A L 29 * * S T A T I S T I C A L L Y S I G N I F I C A N T A T P < 0.01. 220 0.16k 0.14 0 . 1 2 U 0 . 1 0 h 0.08H 0.06 0.04 0.02 k 3 4 5 6 7 8 9 10 DNA C O N C E N T R A T I O N ( U G . / M L . ) A P P E N D I X F I G . |. C A L F T H Y M U S DNA S T A N D A R D C U R V E . 221 RNA C O N C E N T R A T I O N ( U G . / M L . ) A P P E N D I X F I G . I I . Y E A S T RNA S T A N D A R D C U R V E . 0.25 r-P R O T E I N C O N C E N T R A T I O N ( U G . / M L . ) A P P E N D I X F I G . IV. B O V I N E S E R U M A L B U M I N S T A N D A R D C U R V E . 100 90 h 80 h B 70 h 60 J-50 k 40 H 30 f -201 1 1 I I I u_ 0.2 0.4 0.6 0.8 1.0 1.2 R A T I o A/B 1 4 A P P E N D I X F I G . VI. C C H A N N E L S R A T I O Q U E N C H C O R R E C T I O N C U R V E . 

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