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The effects of nutritional factors on behavior and plasma corticosterone concentration in the white leghorn… Gill, Bhupinder Singh 1974

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THE EFFECTS OF NUTRITIONAL FACTORS ON BEHAVIOR AND PLASMA CORTICOSTERONE CONCENTRATION IN THE WHITE LEGHORN CHICKEN BY BHUPINDER SINGH GILL B.V. Sc . &. A . H . , P A N J A B U N I V E R S I T Y , 1962 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN T H E D E P A R T M E N T O F 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 AS 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 T H E 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 A P R I L , 1 9 7 4 I n p r e s e n t i n g t h i s t h e s i s i n p a r t i a l f u l f i l m e n t o f t h e r e q u i r e m e n t s f o r an a d v a n c e d d e g r e e a t t h e U n i v e r s i t y o f B r i t i s h C o l u m b i a , I a g r e e t h a t t h e L i b r a r y s h a l l make i t f r e e l y a v a i l a b l e f o r r e f e r e n c e a n d s t u d y . I f u r t h e r a g r e e t h a t p e r m i s s i o n f o r e x t e n s i v e c o p y i n g o f t h i s t h e s i s f o r s c h o l a r l y p u r p o s e s may b e g r a n t e d b y t h e H e a d o f my D e p a r t m e n t o r by h i s r e p r e s e n t a t i v e s . I t i s u n d e r s t o o d t h a t c o p y i n g o r p u b l i c a t i o n o f t l i i s t h e s i s f o r f i n a n c i a l g a i n s h a l l n o t b e a l l o w e d w i t h o u t my w r i t t e n p e r m i s s i o n . D e p a r t m e n t The U n i v e r s i t y o f B r i t i s h C o l u m b i a V a n c o u v e r 8 , C a n a d a ABSTRACT T H E E F F E C T S OF V A R I O U S D I E T A R Y T R E A T M E N T S ON M E T A B O L I C R E S P O N S E S R E L A T E D TO A D R E N A L F U N C T I O N S H A V E B E E N S T U D I E D IN G R O W I N G W H I T E L E G H O R N C H I C K E N S . B I R D S F E D A D I E T C O N T A I N I N G 20.9% O F P R O T E I N A N D D E F I C I E N T IN L Y S I N E (.51%) S H O W E D H I G H E R P L A S M A C 0 R T I C O S T E R 0 N E C O N C E N T R A T I O N S , I N C R E A S E D A D R E N A L S I Z E , AND I N C R E A S E D O X Y G E N C O N S U M P T I O N C O M P A R E D W I T H B I R D S F E D A D I E T OF S I M I L A R P R O T E I N C O N T E N T AND A D E Q U A T E IN L Y S I N E (1 .11%). B I R D S 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 S H O W E D MORE E X P L O R A T O R Y B E H A V I O U R WHEN P L A C E D ON A G R I D T H A N D I D T H E B I R D S F E D T H E D I E T S U P P L E M E N T E D W I T H L Y S I N E . T H E Y TOOK A L O N G E R T I M E , H O W E V E R , TO L O C A T E F E E D IN A M A Z E . T H I O U R A C I L W I T H H Y P E R V I T A M I N O S I S E , A D M I N I S T E R E D TO B I R D S T H R O U G H T H E D I E T , I N C R E A S E D T H E C O N C E N T R A T I O N O F C O R T I C O S T E R O N E IN T H E P L A S M A . H Y P E R V I T A M I N O S I S E D I D NOT A F F E C T T H E C O N C E N T R A T I O N O F C O R T I C O S T E R O N E IN T H E P L A S M A . T H E C O N C E N T R A T I O N OF C 0 R T I C O S T E R 0 N E WAS S I M I L A R IN T H E P L A S M A OF C H I C K S R E A R E D A T 13 AND 24°C. I I TABLE OF CONTENTS PAGE A B S T R A C T i L I S T OF T A B L E S v L I S T OF F I G U R E S v i L I S T OF A P P E N D I X T A B L E S v i I ACKNOWLEDGEMENTS V I M INTRODUCTION 1 L I T E R A T U R E REVIEW 3 D I E T AND C O R T I C O S T E R O I D S 3 THYROID AND ADRENAL GLAND 10 O X I D A T I V E METABOLISM AND C O R T I C O S T E R O I D S 14 BRAIN AND C O R T I C O S T E R O I D S 16 E X P E R I M E N T A L 18 EXPER I ME NT 1 • 18 E X P E R I M E N T 2 21 E X P E R I M E N T 3 24 E X P E R I M E N T 4 26 E X P E R I M E N T 5 28 RESULTS 32 EXPER i MENT 1 32 E X P E R I M E N T 2 35 E X P E R I M E N T 3 44 E X P E R I M E N T 4 47 E X P E R I M E N T 5 50 D I S C U S S I O N 55 PLASMA C O R T I C O S T E R O N E L E V E L 55 R E S P I R A T I O N RATE 60 ADRENAL WEIGHTS 61 BEHAVIOUR 62 I I I PAGE CONCLUSIONS 64 REFERENCES 66 APPENDIX 78 i v L I S T OF T A B L E S T A B L E P A G E 2 3 1. C O M P O S I T I O N O F E X P E R I M E N T A L D I E T ' 19 1 2 . C O M P O S I T I O N O F D I E T S IN E X P E R I M E N T 2 22 3 . C O M P O S I T I O N OF D I E T S IN E X P E R I M E N T 3 25 4 . C O M P O S I T I O N O F D I E T AND E X P E R I M E N T A L O U T L I N E IN E X P E R I M E N T 4 27 5 . P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN. C H I C K S F E D A 13$ P R O T E I N D I E T W I T H AND W I T H O U T S U P P L E M E N T A R Y L Y S I N E IN E X P E R I M E N T 1 33 6 . P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN C H I C K S F E D D I F F E R E N T D I E T S IN E X P E R I M E N T 2 36 7. R E S P I R A T I O N R A T E OF C H I C K S F E D D I F F E R E N T D I E T S IN E X P E R I M E N T 2 37 8. A D R E N A L W E I G H T S O F C H I C K S F E D D I F F E R E N T D I E T S IN E X P E R I M E N T 2 39 9. B O D Y W E I G H T AND P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN C H I C K S F E D D I F F E R E N T D I E T S IN E X P E R I M E N T 3 45 10. P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN C H I C K S F E D D I F F E R E N T D I E T S M A I N T A I N E D A T 13 C IN E X P E R I M E N T 4 48 11 . P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN C H I C K S F E D D I F F E R E N T D I E T S M A I N T A I N E D A T 24 C IN E X P E R I M E N T 4 49 v L I S T OF F I G U R E S F I G U R E P A G E 1 . S i M P L E M A Z E 30 2 . M A Z E F O R S T U D Y OF E X P L O R A T O R Y B E H A V I O U R IN E X P E R I M E N T 5 31 3 . V A R I A T I O N W I T H T H E A G E I I N P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN C H I C K S F E D A 13% P R O T E I N D I E T W I T H AND W I T H O U T S U P P L E M E N T A R Y L Y S I N E IN E X P E R I M E N T 1 34 4 . GROWTH R A T E S OF C H I C K S F E D D I F F E R E N T D I E T S IN E X P E R I M E N T 2 40 5 . R E L A T I O N S H I P B E T W E E N B O D Y W E I G H T S AND C O R T I C O S T E R O N E C O N C E N T R A T I O N A T TWO W E E K S O F A G E IN C H I C K S F E D D I F F E R E N T D I E T S 41 6 . R E L A T I O N S H I P B E T W E E N B O D Y W E I G H T S AND C O R T I C O S T E R O N E C O N C E N T R A T I O N A T F O U R W E E K S OF A G E IN C H I C K S F E D D I F F E RE NT D I E T S 42 7 . R E L A T I O N S H I P B E T W E E N B O D Y W E I G H T S AND C O R T I C O S T E R O N E C O N C E N T R A T I O N A T S I X W E E K S OF A G E IN C H I C K S F E D D I F F E RE NT D I E T S 43 8. G R O W T H R A T E S OF C H I C K S F E D D I F F E R E N T D I E T S IN E X P E R I M E N T 3 46 9. T I M E S T A K E N BY B I R D S TO D I S C O V E R T H E L O C A T I O N OF F E E D I N E X P E R i M E N T 5 51 10. R E S U L T S O F M O V E M E N T S O F C H I C K S ON G R I D IN E X P E R I M E N T 5 . 52 11 . G R O W T H R A T E S OF C H I C K S F E D L Y S I N E D E F I C I E N T AND L Y S I N E S U P P L E M E N T E D D I E T S IN E X P E R I M E N T 5 53 v I L I S T OF A P P E N D I X T A B L E S A P P E N D I X T A B L E P A G E I. A N A L Y S I S O F V A R I A N C E ( M E A N S S Q U A R E ) O F D A T A F O R P L A S M A C O R T I C O S T E R O N E IN L Y S I N E D E F I C I E N T AND L Y S I N E S U P P L E M E N T E D T R E A T M E N T S . . . 79 I I . A N A L Y S I S . O F V A R I A N C E F O R E L E C T R I C S H O C K R E S P O N S E TO P L A S M A C O R T I C O S T E R O N E IN C H I C K S . . . 80 I I I . A N A L Y S I S O F V A R I A N C E F O R P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N IN C H I C K S A T 45 AND 54 D A Y S O F A G E IN E X P E R I M E N T 2 81 I V . A N A L Y S I S OF V A R I A N C E (M E A N S S Q U A R E ) OF P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N OF C H I C K S IN E X P E R I M E N T 3 82 V . A N A L Y S I S O F V A R I A N C E O F D A T A F R O M E X P E R I M E N T 4 FOR D I E T AND T E M P E R A T U R E I N F L U E N C E ON P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N 83 V I . A V E R A G E B O D Y W E I G H T S 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 AND L Y S I N E S U P P L E M E N T E D D I E T S A T 40 D A Y S O F A G E IN E X P E R I M E N T 5 84 v I I A C K N O W L E D G E M E N T S T H E A U T H O R 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 P R O F E S S O R B . E . M A R C H OF 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 , WHO O R I G I N A L L Y S U G G E S T E D T H E S T U D Y , S U P E R V I S E D T H E R E S E A R C H R E P O R T E D IN AND A D V I S E D IN T H E P R E P A R A T I O N OF T H E M A N U S C R I P T . T H E A C A D E M I C S T I M U L A T I O N AND C O N T I N U E D I N T E R E S T IN T H E P R E P A R A T I O N O F T H E M A N U S C R I P T BY D R . J A C O B B L E L Y OF 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 IS D E E P L Y A P P R E C I A T E D . G R A T E F U L A C K N O W L E D G E M E N T IS A L S O E X T E N D E D B Y , T H E A U T H O R FOR S U G G E S T I N G F I N A L C H A N G E S IN T H E M A N U S C R I P T TO D R . D . B . B R A G G , A S S O C I A T E P R O F E S S O R OF 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 . T H E F A C I L I T I E S R E Q U I R E D F O R T H E C O M P L E T I O N OF T H I S S T U D Y WERE G E N E R O U S L Y P R O V I D E D BY D R . W. D . K I T T S , C H A I R M A N OF 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 , IS G R A T E F U L L Y A C K N O W L E D G E D . T H A N K S A R E E X T E N D E D TO T H E P E R S O N N E L OF T H E P O U L T R Y F A R M AND T H E T E C H N I C A L S T A F F OF T H E P O U L T R Y S C I E N C E L A B O R A T O R Y FOR T H E I R A S S I S T A N C E . V I I I I N T R O D U C T I O N IT IS G E N E R A L L Y C O N S I D E R E D THAT A D R E N A L GLAND A C T I V I T Y IS A S S O C I A T E D WITH S T R E S S . V A R I O U S P H Y S I C A L , C H E M I C A L , N U T R I T I O N A L AND S O C I A L S T R E S S F A C T O R S HAVE B E E N A S S O C I A T E D WITH AN I N C R E A S E IN A D R E N A L C O R T E X S I Z E . I N C R E A S E D I N T A K E OF P R O T E I N HAS B E E N SHOWN TO R E S U L T IN A D R E N A L H Y P E R T R O P H Y . O P P O S I T E CHANGES HAVE B E E N PRODUCED BY P R O T E I N D E P R I V A T I O N . THERE IS A L S O E V I D E N C E THAT P R O T E I N D E F I C I E N C Y I N C H I L D R E N MAY R E S U L T IN A D A P T I V E CHANGES OF H E P A T I C E N Z Y M E S . T H E S E E N Z Y M E S ARE CONCERNED WITH I N T E R M E D I A R Y M E T A B O L I S M , AND ARE C O N S I D E R E D TO BE M E D I A T E D THROUGH THE C O R T I C O S T E R O I D HORMONES. A D M I N I S T R A T I O N OF C E R T A I N A M I N O A C I D S CAN I N F L U E N C E THE F U N C T I O N A L A C T I V I T Y OF A D R E N A L C O R T E X . T H I S IS C O N S I D E R E D DUE TO I N C R E A S E D S E C R E T I O N OF A D R E N O C O R T I C O T R0P H I C HORMONE. S T A R V A T I O N A L S O C A U S E S AN I N C R E A S E I N C O R T I C O S T E R O I D L E V E L S IN THE R A T . T H E R E IS C O N S I D E R E D TO BE A . R E L A T I O N S H I P BETWEEN T H Y R O I D AND A D R E N A L F U N C T I O N . | N M A M M A L S , THE A D R E N A L S OF H Y P E R T H Y R O I D A N I M A L S ARE G E N E R A L L Y IN H Y P E R S E C R E T O R Y S T A T E . THERE IS E V I D E N C E THAT T H Y R O A C T I V E M A T E R I A L S I N THE D I E T A L S O R E S U L T IN A H Y P E R C0R T I C0 I D C O N D I T I O N . INDUCED H Y P O T H Y R O I D I S M HAS B E E N O B S E R V E D TO C A U S E A D R E N A L A T R O P H Y ; AND I N C O N T R A S T , H Y P E R T H Y R O I D I S M MAY PRODUCE A D R E N A L H Y P E R -T R O P H Y IN M A M M A L S . ON THE OTHER H A N D , A D R E N A L H Y P E R T R O P H Y HAS B E E N O B S E R V E D I N H Y P O T H Y R O I D WHITE LEGHORN AND N E W H A M P S H I R E C H I C K E N S . V I T A M I N D E F I C I E N C I E S HAVE B E E N SHOWN TO C A U S E M O R P H O L O G I C A L 1 A L T E R A T I O N S OF T H E A D R E N A L C O R T E X . S C U R V Y WAS T H E F I R S T D E F I C I E N C Y D I S E A S E IN W H I C H G R O S S C H A N G E S IN T H E A D R E N A L G L A N D WERE R E C O G N I Z E D . H Y P E R T R O P H Y AND H E M O R R H A G E I N T O C O R T I C A L T I S S U E H A V E B E E N R E P E A T E D L Y F O U N D IN A N I M A L S D E P R I V E D OF A S C O R B I C A C I D . T H I A M I N E , L I P O I C A C I D , AND P A N T O T H E N I C A C I D A R E F O U N D IN H I G H C O N C E N T R A T I O N IN C O R T I C A L T I S S U E AND A R E C O N S I D E R E D N E C E S S A R Y F O R A D R E N O C O R T I C A L H O R M O N E S E C R E T I O N . V I T A M I N A i s R E P O R T E D TO R E D U C E A D R E N O C O R T I C A L S E C R E T I O N . A L L T H E A B O V E O B S E R V A T I O N S P R O V I D E E V I D E N C E T H A T D I E T A R Y F A C T O R S A R E C O N C E R N E D W I T H A D R E N O C O R T I C A L F U N C T I O N AND i l iHAT T H E F I E L D OF N U T R I T I O N IS C L O S E L Y R E L A T E D TO E N D O C R I N O L O G Y . A L T H O U G H T H E R E H A V E B E E N A N U M B E R O F S T U D I E S ON T H E M A M M A L I A N A D R E N A L G L A N D , L I T T L E R E S E A R C H HAS B E E N R E P O R T E D ON T H E A V I A N A D R E N A L G L A N D IN R E L A T I O N TO D I E T . |N T H E F O L L O W I N G S T U D Y T H E E F F E C T S OF P R O T E I N L E V E L W I T H L Y S I N E L E V E L ; H Y P E R V I T A M I N O S I S E ; AND T H I O U R A C I L ON C O R T I C O S T E R O N E S E C R E T I O N IN C H I C K E N WERE I N V E S T I G A T E D . |N A D D I T I O N , E F F E C T S O F P R O T E I N AND L Y S I N E L E V E L WERE S T U D I E D W I T H R E S P E C T TO A D R E N A L W E I G H T , O X Y G E N C O N S U M P T I O N , E X P L O R A T O R Y A C T I V I T Y , E M O T I O N A L R E A C T I O N , AND GROWTH R E S P O N S E . LITERATURE REVIEW D I E T AND C O R T I C O S T E R O I D S TH E R E IS E V I D E N C E T H A T P R O T E I N - D E F I C I E N C Y IN C H I L D R E N MAY R E S U L T IN A D A P T I V E C H A N G E S O F H E P A T I C E N Z Y M E S C O N C E R N E D W I T H I N T E R M E D I A R Y M E T A B O L I S M AND T H A T T H E S E C H A N G E S A R E M E D I A T E D BY C O R T I C O S T E R O I D H O R M O N E S ( A L L E Y N E AND Y O U N G , 1967) . F U R T H E R I T H A S B E E N SHOWN T H A T T H E M I C R O S O M A L D R U G M E T A B O L I Z I N G E N Z Y M E S O F L I V E R OF G R O W I N G R A T S A R E D E P E N D E N T ON D I E T A R Y I N T A K E OF P R O T E I N AND T O T A L C A L O R I E S ( D I C K E R S O N , E_T _ A L . , 1971) . TH E S E D R U G M E T A B O L I Z I N G E N Z Y M E S O F T H E L I V E R A R E A L S O C O N C E R N E D W I T H M E T A B O L I C D E A C T I V A T I O N O F T H E E N D O G E N O U S S T E R O I D H O R M O N E S AND T H E I R A C T I V I T Y MAY B E I N C R E A S E D BY S T R E S S OR A D M I N I S T R A T I O N OF C O R T I C O S T E R O I D S (D R I E V E R _E_T _AL . , 1966) . I T IS R E P O R T E D T H A T T H E E F F E C T OF D I E T ON T H E A C T I V I T Y O F M E T A B O L I Z I N G E N Z Y M E S OF T H E H E P A T I C D R U G S IS M E D I A T E D T H R O U G H T H E E N D O G E N O U S S T E R O I D H O R M O N E S ( B AS U _ET _AL . , 1971) . T H E A D R E N A L G L A N D IS KNOWN TO BE A F F E C T E D BY T H E P R O T E I N C O N T E N T O F T H E D I E T . AN E X C E S S I V E I N T A K E OF P R O T E I N HAS G E N E R A L L Y B E E N F O U N D TO C A U S E A D R E N A L H Y P E R T R O P H Y R E L A T I V E TO B O D Y W E I G H T . |N S O M E E X P E R I M E N T S , AN I N C R E A S E IN F U N C T I O N A L A C T I V I T Y OF T H E C O R T E X HAS B E E N D E M O N S T R A T E D ( M U N R O , 1964) . O P P O S I T E C H A N G E S A R E I N D U C E D B Y P R O T E I N D E P R I V A T I O N . E V E N A S H O R T P E R I O D OF P R O T E I N D E F I C I E N C Y L E A D S TO A R E D U C T I O N IN A D R E N A L W E I G H T O U T OF P R O P O R T I O N TO L O S S IN B O D Y W E I G H T (MUNRO _ET _ A L . , 1962) . TH I S O C C U R S P A R T I A L L Y T H R O U G H A D I M I N U T I O N IN N U M B E R OF C E L L S P E R G L A N D AND P A R T L Y T H R O U G H A R E D U C T I O N IN A M O U N T S O F C E R T A I N I N D I V I D U A L C O N S T I T U E N T S P E R C E L L . T H E S E E F F E C T S O F D I E T A R Y P R O T E I N L E V E L ON T H E A D R E N A L G L A N D A P P E A R TO BE C A U S E D BY V A R I A T I O N S IN T H E R A T E O F S E C R E T I O N OF A D R E N O C O R T I C O T R 0 P H I C HORMONE ( A C T H ) F R O M T H E P I T U I T A R Y G L A N D (M U N RO _E_T _AL . , 1 9 6 2 ; M U N R O , 1 9 6 4 ) . TH E R E IS A L S O E V I D E N C E T H A T A D M I N I S T R A T I O N O F C E R T A I N A M I N O A C I D S C A N I N F L U E N C E T H E F U N C T I O N A L A C T I V I T Y O F T H E A D R E N A L C O R T E X . TH I S WAS F I R S T S U S P E C T E D WHEN IT WAS SHOWN T H A T U P T A K E OF P H O S P H O R U S - 3 2 BY L I V E R R I B O N U C L E I C A C I D (RNA) IS M A R K E D L Y S T I M U L A T E D BY G I V I N G S I N G L E L A R G E D O S E S O F G L Y C I N E , M E T H I O N I N E OR L E U C I N E TO R A T S (MUNRO AND M U K E R J I , 1 9 6 2 ) , I T WAS C O N C L U D E D T H A T A D M I N I S T R A T I O N OF T H E S E TWO A M I N O A C I D S C A U S E D I N C R E A S E D S E C R E T I O N OF A D R E N O C O R T I C A L H O R M O N E S , W H I C H A R E KNOWN TO S T I M U L A T E L I V E R RNA M E T A B O L I S M ( G O O D L A D AND M U N R O , 1 9 5 9 ) . TH I S E F F E C T O F A M I N O A C I D WAS C O N F I R M E D BY S H O W I N G T H A T A D M I N I S T R A T I O N O F S I N G L E L A R G E D O S E S O F M E T H I O N I N E OR L E U C I N E TO I N T A C T R A T S R A I S E D T H E C O N C E N T R A T I O N O F C 0 R T I C O S T E R O N E IN T H E P L A S M A A L T H O U G H A D M I N I S T R A T I O N O F S E V E R A L O T H E R A M I N O A C I D S IN S I M I L A R D O S E S D I D NOT H A V E T H I S E F F E C T ( M U N R O _ET _AL . , 1 9 6 3 ) . T H I S C O N F I R M S E A R L I E R O B S E R V A T I O N S BY G O T H _ E T _ A L . ( 1 9 5 5 ) ON B L O O D E O S I N O P H I L C O U N T , W H I C H WAS SHOWN TO D I M I N I S H AS A D R E N O C O R T I C A L S E C R E T I O N I N C R E A S E D . A F T E R G I V I N G M E T H I O N I N E OR L E U C I N E TO R A T S , T H E S E A U T H O R S O B S E R V E D A S T R I K I N G R E D U C T I O N IN E O S I N O P H I L P O P U L A T I O N , W H E R E A S O T H E R A M I N O A C I D S P R O D U C E D L E S S S I G N I F I C A N T OR I N S I G N I F I C A N T C H A N G E S . I T IS NOT KNOWN W H E T H E R T H E A C T I O N OF A M I N O A C I D S ON A D R E N O C O R T I C A L S E C R E T I O N D E P E N D S , L I K E T H E A C T I O N OF D I E T A R Y P R O T E I N , ON R E L E A S E O F ACTH FROM P I T U I T A R Y G L A N D . I T WAS F O U N D T H A T C O M P A R I N G C O N T R O L R A T S F E D A P R O T E I N - F R E E D I E T W I T H A N I M A L S R E C E I V I N G C A S E I N OR Z E I N S U P P L E M E N T E D W I T H T R Y P T O P H A N AND L Y S I N E , T H A T T H E P R O T E I N - F E D R A T S HAD C O N S I D E R A B L Y L A R G E R A D R E N A L G L A N D S , T H A T T H E A V E R A G E A D R E N A L C E L L C O N T A I N E D MORE P R O T E I N AND R N A , AND T H A T T H E R E WAS MORE C O R T I C O S T E R O N E IN T H E G L A N D (MUNRO _ET _ A _ . , 1 9 6 5 ) . L L N S U P P L E M E N T E D Z E I N C A U S E D S M A L L E R C H A N G E S IN G L A N D C O M P O S I T I O N , AND F A I L E D TO I N C R E A S E A D R E N A L C O R T I C O S T E R O N E C O N T E N T . F E E D I N G W I T H G E L A T I N HAD NO S I G N I F I C A N T A C T I O N ON G L A N D S I Z E , C O M P O S I T I O N OR C 0 R T I C O S T E R O N E C O N T E N T . T H U S T H E E F F E C T S OF D I F F E R E N T D I E T A R Y P R O T E I N S ON T H E A D R E N A L G L A N D A R E L A R G E L Y D E T E R M I N E D BY T H E I R N U T R I T I V E V A L U E . L A R G E A M O U N T S O F G L Y C I N E , M E T H I O N I N E OR L E U C I N E AS T H E D I E T A R Y S O U R C E OF N I T R O G E N L E D TO E X T E N S I V E I N C R E A S E S IN T O T A L C O R T I C O S T E R O N E C O N T E N T OF A D R E N A L G L A N D W I T H O U T C O R R E S P O N D I N G I N C R E M E N T S IN G L A N D W E I G H T . . A D M I N I S T R A T I O N OF A D R E N O C O R T I C O T R O P H I C HORMONE TO R A T S ON A P R O T E I N - F R E E D I E T C A U S E D I N C R E M E N T S OF S I M I L A R M A G N I T U D E IN G L A N D S I Z E AND IN C O R T I C O S T E R O N E C O N T E N T . F R O M T H I S AND O T H E R E V I D E N C E , I T H A S B E E N C O N C L U D E D T H A T D I E T A R Y P R O T E I N I N F L U E N C E S A D R E N A L S I Z E AND S T E R O I D C O N T E N T T H R O U G H V A R I A T I O N IN R A T E OF A D R E N O C O R T I C 0 T R O P H I C HORMONE S E C R E T I O N . S I N C E A C T I O N O F S I N G L E A M I N O A C I D S ON T H E A D R E N A L G L A N D R E S U L T S IN A C C U M U L A T I O N OF C O R T I C O S T E R O N E W I T H O U T I N C R E A S E IN G L A N D S I Z E , T H I S E F F E C T MAY O C C U R T H R O U G H A M E C H A N I S M O T H E R T H A N A L T E R A T I O N IN AD R E N O C O R T I C O T R O P H I C H O R M O N E S E C R E T I O N . T H E R E WAS NO E V I D E N C E T H A T A C C U M U L A T I O N O F C 0 R T I C O S T E R O N E IN A D R E N A L G L A N D S O F A N I M A L S F E D A N E X C E S S O F G L Y C I N E , M E T H I O N I N E OR L E U C I N E C A U S E D C H A N G E S IN T H E L I V E R C O M P O S I T I O N . E L E V A T I O N OF C O R T I C O S T E R O I D C O N C E N T R A T I O N IN R E S P O N S E TO S T R E S S 6 IN A D U L T R A T S D I F F E R S B E T W E E N A N I M A L S H A N D L E D OR L E F T U N D I S T U R B E D IN I N F A N C Y ( L E V I N E , 1962) . I T HAS B E E N P R O P O S E D T H A T E X P E R I M E N T A L L Y P R O D U C E D S T E R O I D V A R I A T I O N IN I N F A N C Y P E R M A N E N T L Y M O D I F I E S T H E R E G U L A T O R Y M E C H A N I S M S OF H Y P O T H A L M I C - P I T U I T A R Y - A D R E N A L S Y S T E M ( L E V I N E AND M U L L I N S , 1966) . |T IS P O S S I B L E T H A T T H E N U T R I T I O N A L S T A T U S D U R I N G I N F A N C Y MAY A L S O H A V E L A S T I N G E F F E C T S ON T H E S Y S T E M , S I N C E P L A S M A 1 1 - H Y D R O X Y C O R T I C O S T E R O I D C O N C E N T R A T I O N S A R E E L E V A T E D IN M A L N O U R I S H E D C H I L D R E N ( A L L E Y N E AND YOU N G , 1967) . I T IS S U G G E S T E D T H A T E A R L Y U N D E R N U T R I T I O N , IN M A L E R A T S A T L E A S T , D E C R E A S E S T H E I R R E S P O N S I V E N E S S TO S T R E S S F U L S T I M U L A T I O N AS M E A S U R E D BY P L A S M A - 1 1 -H Y D R O X Y C O R T I C O S T E R O I D C O N C E N T R A T I O N S ( A D L A R D A N D S M A R T , 1971, 1972) . I T HAS B E E N D E M O N S T R A T E D T H A T A N I M A L S R E P E A T E D L Y E X P O S E D TO S T R E S S E F F E C T S SHOWED A L O W E R R I S E IN B L O O D C 0 R T I C O S T E R 0 N E L E V E L T H A N D I D C O N T R O L A N I M A L S ( C S A L A Y AND F R E N K L , 1962; C S A L A Y E T A L . , 1965; V E C S E I AND C S A L A Y , 1965) . I T WAS F O U N D T H A T A L C O H O L I S E D R A T S R E S P O N D E D TO A C U T E S T R E S S W I T H A L O W E R R I S E IN B L O O D C 0 R T I C O S T E R O N E T H A N D I D T H E U N T R E A T E D C O N T R O L S (CS A L A Y _ET _AL . , 1 969) . T H I S WAS D E M O N S T R A T E D N O T O N L Y W I T H A L C O H O L , W H I C H WAS R E S P O N S I B L E F O R A D A P T A T I O N , B U T A L S O W I T H A Q U I T E D I F F E R E N T S T R E S S O R , V I Z . S T R E N O U S S W I M M I N G E X E R C I S E F O R O N E H O U R . S L N C E T H E A L C O H O L I Z E D R A T S R E S P O N D E D MORE I N T E N S I V E L Y T H A N U N T R E A T E D C O N T R O L S TO E X O G E N O U S A C T H , I T IS S U P P O S E D T H A T I N H I B I T I O N OF E N D O G E N O U S A C T H S E C R E T I O N MAY P L A Y A R O L E IN T H E P H E N O M E N O N . S O C I A L S T R E S S HAS B E E N F O U N D TO M O D I F Y T H E R E S P O N S E OF C H I C K E N S TO I N F E C T I O U S D I S E A S E . C H I C K E N S M A I N T A I N E D IN A MORE S T R E S S F U L S O C I A L E N V I R O N M E N T ( G R O S S AND C O L M A N O , 1969), WERE MORE S U S C E P T I B L E TO V I R A L I N F E C T I O N S AND MORE R E S I S T A N T TO B A C T E R I A L I N F E C T I O N S T H A N C H I C K E N S M A I N T A I N E D IN A L E S S S T R E S S F U L S O C I A L E N V I R O N M E N T . H L G H L E V E L S O F P L A S M A C O R T I C O S T E R O N E WERE SHOWN TO BE P O S I T I V E L Y C O R R E L A T E D W I T H R E S I S T A N C E TO E S C H E R I CHI A C O L I I N F E C T I O N ( G R O S S AND C O L M A N O , 1 9 6 7 ) . I N J E C T I O N S OF C H I C K E N S W I T H C O R T I C O S T E R O N E R E S U L T E D IN I N C R E A S E D R E S I S T A N C E TO E . C O L I I N F E C T I O N ( G R O S S AND C O L M A N O , 1 9 7 0 ) . C H E M I C A L S W H I C H B L O C K A D R E N A L P R O D U C T I O N OF C O R T I C O S T E R O N E R E S U L T E D IN I N C R E A S E D R E S I S T A N C E TO V I R A L I N F E C T I O N S AND D E C R E A S E D R E S I S T A N C E TO E . C O L I I N F E C T I O N ( C O L M A N O AND G R O S S , 1 9 7 1 ) . BROWN AND N E S T O R ( 1 9 7 0 ) R E P O R T E D T H A T T U R K E Y S C O U L D B E S E L E C T E D F O R P L A S M A C 0 R T I C O S T E R 0 N E R E S P O N S E TO C O L D S T R E S S . T H E Y F O U N D T H A T M A J O R S E L E C T I V E M O D I F I C A T I O N WAS ON T H E A B I L I T Y O F A D R E N A L S TO R E S P O N D TO A C T H . G R O S S AND C O L M A N O ( 1 9 7 1 ) F O U N D T H A T C H I C K E N S S E L E C T E D F O R A LOW P L A S M A C 0 R T I C O S T E R O N E L E V E L AS A R E A C T I O N TO S T R E S S WERE MORE R E S I S T A N T TO M A R E K ' S D I S E A S E AND M Y C O P L A S M A G A L L I S E P T I C U M I N F E C T I O N , P R O D U C E D MORE A N T I B O D Y A N D WERE MORE S U S C E P T I B L E TO E . C O L I I N F E C T I O N AND WERE MORE R E S P O N S I V E TO A D R E N A L B L O C K I N G A G E N T S T H A N WERE C H I C K E N S S E L E C T E D FOR A H I G H P L A S M A C O R T I C O S T E R O N E R E A C T I O N . S T A R V A T I O N C A U S E S AN I N C R E A S E IN P L A S M A C O R T I C O S T E R O I D L E V E L S IN A D U L T R A T S ( B O U L O U A R D , 1 9 6 3 ) . AN E F F E C T O F U N D E R N U T R I T I O N ON A D R E N O C O R T I C A L F U N C T I O N WAS NOT TO BE E X P E C T E D M U C H B E F O R E 1 5 D A Y S O F A G E S I N C E O N L Y A T T H I S T I M E DO M A R K E D R E S P O N S E S TO C O R T I C O T R O P H I N ( A C T H ) AND S T R E S S A P P E A R ( A L L E N AND K E N D A L L , 1 9 6 7 ; L . E V I N , 1 9 6 8 ) . |T WAS P R O P O S E D T H A T A L T E R A T I O N IN C O R T I C O S T E R O I D L E V E L S IN I N F A N C Y M A Y P E R M A N E N T L Y M O D I F Y T H E R E G U L A T I O N OF T H E H Y P O T H A L A M I C - P I T U I T A R Y - A D R E N A L S Y S T E M ( L E V I N E AND M U L L I N S , 1 9 6 6 ; A D L A R D AND S M A R T , 1 9 7 1 ) . THERE IS MUCH C I R C U M S T A N T I A L E V I D E N C E THAT ADRENOCORTICO -S T E R O I D S OF THE C O R T I S O L TYPE ARE INVOLVED IN CONTROL OF METABOLISM IN THE STARVED MAMMALS. MANY OF R E L E V A N T EXPERIMENTS CONCERN THE E F F E C T S OF BOTH ADRENALECTOMY AND TREATMENT WITH EXOGENOUS ADRENOCORTICAL HORMONES ON M O B I L I Z A T I O N OF T I S S U E P R O T E I N ( W H I T E , 1947; D ' ADGELO _ET A L . , 1948; ENGEL E J _AL . , 1949) . THESE RESULTS POINT TO THE IMPORTANCE OF C O R T I S O L AND C O R T I C O S T E R0N E IN PROVIDING AMINO ACIDS BOTH AS AN ENERGY SOURCE AND FOR REASSEMBLY AS PROTE IN ELSEWHERE IN THE BODY. EXPERIMENTS HAVE ALSO DEMONSTRATED THAT C O R T I S O L HAS THE PROPERTY OF INDUCING S P E C I F I C L I V E R ENZYMES CONCERNED WITH AMINO ACID METABOLISM AND G L U C O N E O G E N E S I S (KNOX AND A V E R B A C H , 1955; KENNY, 1962; GREENGARD AND F E I G E L S O N , 1962; S I E AND F I S H M A N , 1964; WEBER AND S l N G H A L , 1964) . B E L L A M Y _ET _AL . (1968) FOUND T H A T , A F T E R THREE DAYS OF S T A R V A T I O N IN R A T S , CARCASS AND L I V E R LOST 20 AND 40$ R E S P E C T I V E L Y OF THE I N I T I A L WEIGHT. THE FAT BODIES DECREASED BY 50$ IN TWO DAYS OF S T A R V A T I O N . THERE WAS NO CHANGE IN P R O T E I N CONTENT OF L I V E R AND C A R C A S S . PRETREATMENT WITH METAP I RONE OR P R O P Y L T H I O U R A C I L I N H I B I T E D THE LOSS OF WEIGHT IN CARCASS AND L I V E R BY 70 AND 20$ R E S P E C T I V E L Y . THYROXINE RESTORED THE NORMAL WEIGHT LOSS IN S T A R V E D , M E T A P I R O N E -TREATED A N I M A L S . REMOVAL OF FOOD A B O L I S H E D THE DIURNAL RHYTHM, AND ILL HE RE WAS A LARGE INCREASE IN THE S T E R O I D C O N C E N T R A T I O N . PRETREATMENT WITH ME TAP I RONE PREVENTED T H I S R I S E IN C0R T I C O S T E R O N E . THERE WAS AN INCREASE IN THE A C T I V I T Y OF TYROS I N E - C T - O X O G L U T A R A T E TRANSAMINASE IN THE L I V E R OF STARVED R A T S , AND A C T I V I T Y OF THE ENZYME F L U C T U A T E D IN L I N E WITH THE V A R I A T I O N S IN PLASMA C0R T I C O S T E R O N E . |N C O N T R A S T , THE T O T A L A C T I V I T Y OF G L U T A M A T E - P Y R U V A T E T R A N S A M I N A S E I N C R E A S E D G R A D U A L L Y D U R I N G T H R E E D A Y S O F S T A R V A T I O N ( F O U R - F O L D I N C R E A S E ) . E V I D E N C E HAS B E E N P R E S E N T E D S H O W I N G T H A T C O N S U M P T I O N OF H I G H P R O T E I N D I E T S BY Y O U N G C H I C K S I S A C C O M P A N I E D BY M A R K E D R E D U C T I O N IN L I V E R V I T A M I N A R E S E R V E S AND I N C R E A S E IN A D R E N A L S I Z E ( S T O E W S A N D AND S C O T T , 1 9 6 1 , 1 9 6 4 ) . A C C O R D I N G TO S E L Y E ( 1 9 4 7 ) A H Y P E R T R O P H I E D AND H Y P E R S E C R E T O R Y A D R E N A L C O R T E X IS A S S O C I A T E D W I T H T H E F I R S T S T E P OF AN O R G A N I S M ' S R E A C T I O N TO S T R E S S . V I T A M I N A N U T R I T I O N HAS B E E N I M P L I C A T E D AS AN A D J U N C T IN P R O T E C T I O N O F A N I M A L S A G A I N S T A V A R I E T Y OF S T R E S S E S S U C H AS C O L D E X P O S U R E , ( E R S H O F F , 1 9 5 2 ; O D A G I R I AND K O Y A N G I , 1 9 6 1 ) , X - I R R A D I A T I ON ( E R S H O F F , 1 9 5 2 ) , AND I N F E C T I O U S D I S E A S E S ( G R E E N AND M A L L A N B Y , 1 9 2 8 ; B O Y N T O N AND B R A D F O R D , 1 9 3 1 ; P A N D A AND C O M B S , 1 9 6 3 ) . W O L F E_T _ A L . ( 1 9 5 8 ) AND V A N D Y K E _ET _ A L . ( 1 9 5 7 ) P R E S E N T E D E V I D E N C E T H A T V I T A M I N A D I R E C T L Y I N F L U E N C E S HORMONE P R O D U C T I O N OF T H E A D R E N A L C O R T E X TO S U C H AN E X T E N T T H A T S E V E R E V I T A M I N A D E F I C I E N C Y WAS C O N S I D E R E D TO C O N S T I T U T E " C H E M I C A L A D R E N A L E C T O M Y 1 ' IN FLIHE R A T . G L I C K ( 1 9 6 3 ) O B S E R V E D A R E D U C E D H E T E R O P H I L C O U N T IN B L O O D OF V I T A M I N A D E F I C I E N T C H I C K S W H I C H I N D I C A T E D A R E D U C E D F U N C T I O N I N G OF A D R E N A L C O R T E X ( H U B L E , 1 9 5 5 ) . E V I D E N C E A L S O A P P E A R E D I N D I C A T I N G T H A T A D R E N A L C O R T I C A L H O R M O N E S MAY I N F L U E N C E V I T A M I N A M E T A B O L I S M . C L A R K AND C O L B U R N ( 1 9 5 5 ) R E P O R T E D T H A T L A R G E D O S E S O F C O R T I S O N E A C E T A T E , I N J E C T E D I N T O N O R M A L OR A D R E N A L E C T O M I Z E D R A T S , C A U S E D A D E P L E T I O N OF L I V E R V I T A M I N A . M C G I L L I V R A Y ( 1 9 6 1 ) A L S O O B S E R V E D T H A T F O U R MG OF C O R T I S O N E I N J E C T E D D A I L Y I N T O R A T S C A U S E D A R E L E A S E O F V I T A M I N A A L C O H O L IN T H E B L O O D . I N J E C T E D ACTH P R O D U C E D T H E S A M E R E S U L T S TO A L E S S E R E X T E N T . T H Y R O I D AND A D R E N A L G L A N D T H E A D R E N A L G L A N D HAS B E E N R E P O R T E D TO B E C O N S I D E R A B L Y E N L A R G E D AS A R E S U L T O F F E E D I N G TRI - I O D O T H Y R O N I NE OR T H Y R O X I N E ( S L N H A AND S C H M I D T , 1 9 7 0 ; D E A R I E AND G R E E P , 1 9 4 7 ; W A L L A C H AND R E I N E K E , 1 9 4 9 ) . T H E R E S P O N S E MAY H A V E R E S U L T E D FROM I N C R E A S E D ACTH S E C R E T I O N . S O M E I N C O M P L E T E R E S U L T S OF ACTH A S S A Y OF P I T U I T A R Y G L A N D O F T H E S E A N I M A L S I N D I C A T E D S U C H A T R E N D . H I L T O N J_T _AJ_. ( 1 9 6 2 ) H A V E R E P O R T E D I N C R E A S E D ACTH S E C R E T I O N IN MAN D U E TO H Y P E R T H R O I D I S M . S T E I N E T Z AND B E E C H ( 1 9 6 3 ) D I D NOT O B S E R V E A N Y I N C R E A S E IN P I T U I T A R Y ACTH IN R A T S IN A M I L D C O N D I T I O N OF H Y P E R T H Y R O I D I S M B U T D I D O B S E R V E AN I N C R E A S E IN P L A S M A C O R T I C O S T E R O I D O F T H Y R0 I D - T R E A T E D A N I M A L S F O L L O W I N G ACTH I N J E C T I O N , A L T H O U G H T H E Y A T T R I B U T E D T H I S I N C R E A S E TO A R E D U C E D V O L U M E OF D I S T R I B U T I O N OF T H E C O R T I C O I D S . |N T H E S E C O N D S E R I E S OF E X P E R I M E N T S OF T H I S S T U D Y , P L A S M A C O R T I C O S T E R O I D C O N C E N T R A T I O N WAS H I G H E R WHEN T H Y R O A C T I V E M A T E R I A L WAS A D M I N I S T E R E D TO R A T S A T D I F F E R E N T S T A G E S OF L A C T A T I O N ( S I N H A AND S C H M I D T , 1 9 7 0 ) . IT HAS A L S O B E E N D E M O N S T R A T E D T H A T H Y P E R T H R O I D I S M G R E A T L Y A C C E L E R A T E S T H E H E P A T I C I N A C T I V A T I O N OF A D R E N A L S T T E R O I D S ( S C H R I E F E R S , 1 9 6 7 ) . F U R T H E R M O R E , W A L L A C H AND R E I N E K E ( 1 9 4 9 ) H A V E SHOWN T H A T T H E A D R E N A L G L A N D OF A H Y P E R T H Y R O I D R A T I S I N A S T A T E O F H Y P E R S E C R E T I O N . T H U S , I T A P P E A R S T H A T A N I M A L S F E D T H Y R O A C T I V E M A T E R I A L D E V E L O P A H Y P E R C O R T I C 0 ID C O N D I T I O N . T H I S C A N P A R T I A L L Y A C C O U N T F O R L A C T A T I O N A L I N S U F F I C I E N C Y S I N C E B O T H H Y P E R -AND H Y P O - A D R E N A L HORMONE A C T I V I T Y A D V E R S E L Y A F F E C T L A C T A T I O N ( C O W I E AND F O L L Y , 1 9 6 1 ) . W H E T H E R H Y P E R T H Y R O I D I S M A L S O I N C R E A S E S C O R T I -C O S T E R O I D - BI ND I NG G L O B U L I N ( C B G ) A C T I V I T Y AND R E N D E R S T H E A D D I T I O N A L C O R T I C O I D S U N A V A I L A B L E FOR A C T I O N IS NOT K N O W N . P R O L A C T I N AND A D R E N A L H O R M O N E S A R E NOT T H E O N L Y H O R M O N E S A F F E C T E D BY H Y P E R T H Y R O I D I S M . T H Y R O I D H O R M O N E S A R E M E T A B O L I C H O R M O N E S H A V I N G A W I D E R A N G E OF A C T I O N . T H E P O S S I B I L I T Y T H A T A V E R Y H I G H D E G R E E O F H Y P E R T H Y R O I D I S M C A N A D V E R S E L Y A F F E C T M A N Y OF T H E O T H E R E N D O C R I N E AND M E T A B O L I C S Y S T E M S E S S E N T I A L FOR N O R M A L L A C T A T I O N M U S T A L S O B E C O N S I D E R E D . TH E R E P O R T S O F T H E E F F E C T S OF T H Y R O I D H O R M O N E S ON M I L K P R O D U C T I O N IN R A T S A R E C O N F L I C T I N G . S E V E R A L W O R K E R S ( G R O S V E N O R AND T U R N E R , 1959; VO N BE R S W O R D T EJT A L . . , 1960) R E P O R T E D T H A T T H Y R O X I N E I N J E C T I O N S I N C R E A S E M I L K P R O D U C T I O N AND L I T T E R W E I G H T G A I N O F L A C T A T I N G R A T S . MORE R E C E N T L Y , H O W E V E R , TR A V I s _ET _AL . (1964) F O U N D T H A T F E E D I N G OF E I T H E R D R I E D B O V I N E T H Y R O I D G L A N D W I T H P A R A T H Y R O I D G L A N D OR C A L F G U L L E T T R I M M I N G W I T H T H Y R O I D AND P A R A T H Y R O I D G L A N D S , OR A D I E T C O N T A I N I N G TRI - I O D O T H Y R O N I NE AND T H Y R O X I N E TO R A T S OR M I N K R E D U C E D T H E N U M B E R O F Y O U N G B O R N , I N C R E A S E D M O R T A L I T Y O F Y O U N G , AND R E D U C E D T H E R A T E O F B O D Y W E I G H T G A I N OF T H E L I V E Y O U N G . P U R S U I N G T H I S F U R T H E R , S C H M I D T AND M O G E R (1967) C O N C L U D E D T H A T GROWTH OF T H E Y O U N G WAS I N H I B I T E D B Y I M P A I R M E N T O F M A T E R N A L L A C T A T I O N . H Y P O T H Y R O I D I S M , I N D U C E D BY T H I O U R A C I L , HAD NO E F F E C T ON T H E A D R E N A L W E I G H T OF F O U R - W E E K OLD C H I C K S O V E R A 3 TO 17 D A Y P E R I O D , A L T H O U G H B O T H B U R S A A N D S P L E E N R E G R E S S E D I N D I C A T I N G A P O S S I B L E I N C R E A S E IN S T E R O I D S E C R E T I O N . |N B I R D S OF S A M E A G E G R O U P , A D D I T I O N O F T H Y R O P R O T E I N TO T H E FOOD TO P R O M O T E H Y P E R T H Y R O I D I S M D I D NOT P R O D U C E I N C R E A S E D A D R E N A L M A S S U N T I L 12 D A Y S L A T E R ( G A R R E N AND S H A F F N E R , 1956) . A W I D E V A R I E T Y O F S T R E S S S T I M U L I E M P L O Y E D IN YOUNG C H I C K E N S H A V E P R I M A R I L Y R E S U L T E D IN A D R E N A L E N L A R G E M E N T W H I C H WAS A C C O M P A N I E D BY AN I N V O L U T I O N O F L Y M P H A T I C T I S S U E . C O L D IS P E R H A P S T H E M O S T COMMON C O N D I T I O N OF S T R E S S E N C O U N T E R E D BY L I V I N G O R G A N I S M . S U R V I V A L IN E X T R E M E C O L D F O R M O S T A N I M A L S IS D E P E N D E N T U P O N T H E O C C U R R E N C E OF A N U M B E R OF P H Y S I O L O G I C A L A D J U S T M E N T S . A s R E P O R T E D BY H O F F M A N AND S H A F F N E R ( 1 9 5 0 ) , T H E M E T A B O L I C R A T E OF T H Y R O I D A C T I V I T Y OF C H I C K E N S I N C R E A S E D F O L L O W I N G E X P O S U R E TO C O L D . N U M E R O U S I N V E S T I G A T O R S H A V E SHOWN T H A T E X P O S U R E TO C O L D C A U S E S M A N Y OF T H E P H Y S I O L O G I C A L R E S P O N S E S W H I C H A R E C O M M O N L Y O B S E R V E D F O R M A N Y O T H E R F O R M S O F S T R E S S S T I M U L I . T H E C H A R A C T E R I S T I C A D R E N A L H Y P E R T R O P H Y ( B R O L I N , 1 9 4 6 ) O C C U R S P A R T I C U L A R L Y I F T H E A N I M A L S H A V E B E E N S E N S I T I Z E D TO S T R E S S B Y S T A R V A T I O N ( S E L Y E , 1 9 3 6 ; L E B L O N D AND DU G A L , 1 9 4 3 ) . A T H Y M I C - L Y M P H A T I C I N V O L U T I O N A L S O O C C U R S AS R E P O R T E D BY 1_E B L 0 ND _E_T _AL . ( 1 9 3 9 ) . IN D U C E D H Y P O T H Y R O I D I S M HAS B E E N O B S E R V E D TO C A U S E A D R E N A L A T R O P H Y IN T H E R A T ( L E B L O N D AND H O F F , 1 9 4 4 ; B A U M A N N AND M A R I N E , 1 9 4 5 ; AND D E A R I E AND G R E E P , 1 9 4 7 ) ; T H E C A T (MCC L O S K Y E_T _AL . , 1 9 4 7 ) ; AND G U I N E A P I G ( F L O R E N T I N _ET _ A L . , 1 9 4 7 , AND M C Q U I L L A N AND T R I K O J U S , 1 9 4 6 ) . |N C O N T R A S T , H Y P E R T H Y R O I D I S M R E S U L T S IN A D R E N A L H Y P E R T R O P H Y ( S W A N N , 1 9 3 9 ; M I L L E R AND R I D D L E , 1 9 3 9 ; KA D E N E_T A _ L . , 1 9 3 7 ; DU R E Y , 1 9 4 9 ; AND S E L Y E , 1 9 4 9 ) . S E E M I N G L Y I N C O N F L I C T W I T H T H E R E S U L T S O B T A I N E D W H E R E M A M M A L S WERE U S E D , M O R R I S ( 1 9 5 3 ) R E P O R T E D A D R E N A L H Y P E R T R O P H Y IN WHITE L E G H O R N C H I C K E N S IN W H I C H H Y P O T H Y R O I D I S M WAS I N D U C E D E I T H E R B Y A D M I N I S T E R I N G T H I O U R A C I L A T 0.1 P E R C E N T O F T H E D I E T OR B Y T H Y R O I D E C T O M Y . AN A D R E N A L H Y P E R T R O P H Y O F T H I S N A T U R E WAS R E P O R T E D BY GA R R E N AND S H A F F N E R ( 1 9 5 2 ) IN NEW HA M P S H I R E C H I C K E N S G I V E N T H I O U R A C I L IN T H E F E E D A T A L E V E L OF 0.2 P E R C E N T . R E D U C E D A T M O S P H E R I C P R E S S U R E S H A V E B E E N F O U N D BY S E V E R A L I N V E S T I G A T O R S TO C A U S E A D R E N A L H Y P E R T R O P H Y AND L Y M P H A T I C I N V O L U T I O N (AR M S T R O N G AND H E I M , 1 9 3 8 ; THORN _E_T _ A L . , 1 9 4 2 j L A N G L E Y AND C L A R K , 1 9 4 2 ; AND DO H A N , 1 9 4 2 ) . AN O B S E R V A B L E C H A N G E IN C O R T I C A L L I P I D S WAS N O T E D BY DARROW AND S A R A S O N ( 1 9 4 4 ) IN T H E A D R E N A L G L A N D S OF R A T S S U B J E C T E D TO S I M U L A T E D A L T I T U D E O F 2 0 , 0 0 0 TO 2 5 , 0 0 0 F E E T FOR O N L Y TWO H O U R S . O X I D A T I V E M E T A B O L I S M AND C O R T I C O S T E R O I D S S E V E R A L I N V E S T I G A T O R S H A V E R E P O R T E D E F F E C T S OF G L U C O C O R T I C O I D S ON O X I D A T I V E M E T A B O L I S M ( C L A R K AND P E S C H , 1 9 5 6 ; G A L L A G H E R , 1 9 5 9 ; G O E T S C H AND M C D O N A L D , 1 9 6 2 ) . T H E S E E F F E C T S S E E M TO V A R Y D E P E N D I N G U P O N T H E T I S S U E U S E D AND T H E D U R A T I O N OF T R E A T M E N T . P R E V I O U S WORK H A S SHOWN T H A T S I N G L E I N J E C T I O N S OF P R E D E N I S O L 0 N E OR H Y D R O C O R T I S O N E I N C R E A S E D O X Y G E N U P T A K E IN L I V E R H O M O G E N A T E S , W H E R E A S C H R O N I C I N J E C T I O N S D E C R E A S E D O X Y G E N U P T A K E ( G O E T S C H AND M C D O N A L D , 1 9 6 2 ; G O E T S C H , 1 9 6 3 ) . C L A R K AND P E S C H ( 1 9 5 6 ) F O U N D T H A T C H R O N I C A D M I N I S T R A T I O N OF C O R T I S O N E L O W E R E D O X Y G E N U P T A K E , B U T HAD NO E F F E C T ON O X I D A T I V E P H O S P H O R Y L A T I O N OF NORMAL R A T L I V E R M I T O C H O N D R I A . |N C O N T R A S T K E R P P O L A ( 1 9 6 0 ) R E P O R T E D T H A T C H R O N I C I N J E C T I O N S O F C O R T I S O N E I N H I B I T E D O X I D A T I V E P H O S P H O R Y L A T I O N . S T U D I E S IN V I T R O I N D I C A T E D T H A T H Y D R O C O R T I S O N E I N H I B I T E D O X I D A T I V E M E T A B O L I S M IN R A T L I V E R M I T O C H O N D R I A ( G A L L A G H E R , 1 9 5 9 ) . S T I C K L A N D ( 1 9 6 2 ) R E P O R T E D NO A L T E R A T I O N IN P H O S P H O R U S TO O X Y G E N (P :0 ) R A T I O S OF L I V E R M I T O C H O N D R I A OR H O M O G E N A T E S F O L L O W I N G A D R E N A L E C T O M Y . |N C O N T R A S T L L L J E R O O T AND H A L L ( 1 9 6 5 ) F O U N D T H A T A D R E N A L E C T O M Y D E C R E A S E D P :0 R A T I O IN R A T L I V E R M I T O C H O N D R I A AND T H E A D D I T I O N O F C O R T I S O N E IN V I T R O I N C R E A S E D L I V E R M I T O C H O N D R I A L P :0 R A T I OS . I T WAS F O U N D T H A T C O R T 1 C O S T E R O N E T R E A T M E N T E N H A N C E D O X I D A T I V E M E T A B O L I S M IN R A T L I V E R H O M O G E N A T E S AND I N C R E A S E D ATP F O R M A T I O N AND P :0 R A T I O S IN R A T L I V E R M I T C H O N D R I A U S I N G M A L A T E AS A S U B S T R A T E . T H E R E S U L T S I N D I C A T E D T H A T C 0 R T I C O S T E R 0 N E HAS AN E F F E C T ON M I T O C H O N D R I A L R E S P I R A T I O N N E A R T H E NAD - F L A V O P R 0 T E I N P O R T I O N OF E L E C T R O N T R A N S P O R T C H A I N (GE R A L D _E_T _ A L . , 1968) D A I L Y R H Y T H M S IN F E E D I N G ( R I C H T E R , 1927; S I E G E L AND S T U C K E Y , 1 947; G I L B E R T AND G A M E S , 1956; S I E G E L , 1961; IBA L A G U R A AND C O S C I N A , 1968) AND A C T I V I T Y ( R I C H T E R , 1922; 1927; BR O W M A N , 1937; B O L L E S AND O G I L V I E , 1966) IN R A T S A R E W E L L R E C O R D E D , T O G E T H E R W I T H A N U M B E R OF D A I L Y R H Y T H M S OF P H Y S I O L O G I C A L P R O C E S S E S ( H A R K E R , 1957). L I T T L E I N F O R M A T I O N IS A V A I L A B L E C O N C E R N I N G E N D O C R I N E I N F L U E N C E S ON D A I L Y R H Y T H M S OF M E T A B O L I C A C T I V I T I E S IN V E R T E R B R A T E S A L T H O U G H A N U M B E R OF I N V E S T I G A T I O N S (HA R K E R , 1 964; F U L L E R AND SN O D D Y , 1 968; B E L L A M Y , E T A L . , 1968) H A V E E X A M I N E D D A I L Y R H Y T H M S IN L E V E L OF P L A S M A C O R T I C O S T E R O N E IN T H E N O R M A L R A T IN R E L A T I O N TO F E E D I N G AND G R O W T H . T H E S U G G E S T I O N WAS M A D E T H A T S K E L E T A L M U S C L E AND L I V E R WERE G R O W I N G O U T OF P H A S E BY 12 H , AND IT WAS P O S T U L A T E D T H E C O R T I C O S T E R O N E E X E R T E D AN I N H I B I T O R Y I N F L U E N C E ON GROWTH IN T H E F O R M E R , AND S T I M U L A T E D GROWTH IN T H E L A T T E R T I S S U E . T H E P O S T U L A T E T H A T G E N E R A L B O D Y GROWTH C O N F O R M E D TO A D I U R N A L P A T T E R N , B E I N G G R E A T E S T IN T H E D A R K , WAS S U B S T A N T I A T E D BY S T U D I E S ON D I U R N A L R H Y T H M OF W A T E R AND P O T A S S I U M R E T E N T I O N IN T H E R A T ( B E L L A M Y , E T A L . , 1969) . D A I L Y R H Y T H M S IN O X Y G E N C O N S U M P T I O N WERE D E M O N S T R A T E D IN L I V E R AND S K E L E T A L M U S C L E S OF M A L E R A T S . T H E E F F E C T OF A D R E N A L E C T O M Y U P O N R H Y T H M S WAS E X A M I N E D . T H I S I N D I C A T E D T H A T A D R E N A L A C T I V I T Y D O E S NOT D E T E R M I N E T H E P H A S I N G OF T H E R H Y T H M S B U T D O E S C O N T R O L T H E B A S A L L E V E L OF O X Y G E N U P T A K E ( G O U L D I N G AND B E L L A M Y , 1971) . BR A I N AND C O R T I C O S T E R O I D HO R M O N A L C O N T R O L OF E N Z Y M E L E V E L S IS Q U I T E COMMON IN M O S T M A M M A L I A N T I S S U E S . S O B E L AND E C K S T E I N ( 1 9 5 9 ) R E P O R T E D T H A T B R A I N S U C C I N A T E D E H Y D R O G E N A S E A C T I V I T Y IS I N C R E A S E D BY O V A R I E C T O M Y AND D E C R E A S E D BY T E S T O S T E R O N E A D M I N I S T R A T I O N . T H Y R O I D H O R M O N E S I N F L U E N C E T H E D E V E L O P M E N T OF S E V E R A L E N Z Y M E S IN T H E B R A I N OF N E O N A T A L R A T S (HA M B U R G H AND F L E X N E R , 1 9 5 7 ; G E E L AND T I M I R A S , 1 9 6 7 ; B A L A Z S , E T A L . , 1 9 6 7 ; ROB i N S O N __r _AL . , 1 9 6 7 ) . TH Y R O I D H O R M O N E S H A V E NO S U C H I N F L U E N C E , H O W E V E R , IN A D U L T B R A I N ( L E E AND L A R D Y , 1 9 6 5 ) . S I M I L A R L Y P R O T E I N S Y N T H E S I S IS S T I M U L A T E D BY T H Y R O X I N E IN Y O U N G B U T NOT IN A D U L T R A T B R A I N ( G E L B E R _ET _ A L . , 1 9 6 4 ) . FO L L O W I N G H Y P O P H Y S E C T O M Y OR A D R E N A L E C T O M Y G L Y C E R O L P H O S P H A T E D E H Y D R O G E N A S E ( G P D H ) A C T I V I T Y D E C R E A S E D E X P O N E N T I A L L Y IN T H E C E R E B R A L H E M I S P H E R E S AND B R A I N S T E M OF A D U L T M A L E R A T S (jEAN DEVELLIS AND I N G L I S H , 1 9 6 8 ) . M A L A T E D E H Y D R O G E N A S E , L A C T A T E D E H Y D R O G E N A S E AND M I T O C H O N D R I A L G L Y C E R O P H O S P H A T E D E H Y D R O G E N A S E A C T I V I T I E S R E M A I N E D U N C H A N G E D . I N J E C T I O N S OF A A D R E N O C O R T I C O T R O P H I C HORMONE OR C O R T I S O L IN A D REN A L E C T O M I Z E D R A T S R E S T O R E D G P D H A C T I V I T Y . T H Y R O I D E C T O M Y OR G O N A D E C T O M Y HAD NO E F F E C T ON G P D H A C T I V I T Y . L I V E R G P D H WAS N O T D I M I N I S H E D B Y H Y P O P H Y S E C T O M Y OR A D R E N A L E C T O M Y AND AS MUCH AS B R A I N G P D H BY H Y P O P H Y S E C T O M Y . |N Y O U N G R A T S G P D H D E V E L O P M E N T A L I N C R E A S E IN A C T I V I T Y IS S P E C I F I C A L L Y R E G U L A T E D BY C O R T I S O L ( A N D P R O B A B L Y C L O S E L Y R E L A T E D C O R T I C O S T E R O I D ) . T R Y P T O P H A N H Y D R O X Y L A S E IS T H E F I R S T O F TWO E N Z Y M E S R E S P O N S I B L E F O R B I O S Y N T H E S I S O F S E R O T O N I N F R O M T R Y P T O P H A N . |T IS F O U N D IN R E G I O N S OF B R A I N W H E R E , AS J U D G E D B Y S P E C I F I C H I S T O F L U O R E S C E N C E , T H E C E L L B O D I E S AND E N D I N G S OF N E U R O N E S C O N T A I N I N G T H E A M I N E S A R E L O C A T E D ( D A H L S T R O M AND F U X E , 1 9 6 4 ) . S E R O T O N I N MAY F U N C T I O N AS A C H E M I C A L S Y N A P T I C T R A N S M I T T E R / 17 AND R E G U L A T O R Y M E C H A N I S M F O R S E R O T O N I N F O R M A T I O N AND U T I L I Z A T I O N A R E B E I N G I N V E S T I G A T E D (C H A S E _ET _AL . , 1 9 6 8 ) . P O S S I B L E E F F E C T S OF A D R E N A L S T E R O I D S ON S E R O T O N I N C O N C E N T R A T I O N H A V E B E E N S T U D I E D ( D E M A I O , 1 9 5 9 ] Y E H _ E T _ A L . , 1 9 5 9 ; G A R A T T I N I _ET _ A L . , 1 9 6 0 ; PU T AND M E D U S K I , 1 9 6 2 ; SOF'ER _ET _ A L . , 1 9 6 2 ; P F E I F E R _ET M.., 1 9 6 3 ) . THE R O L E OF C O R T I C O S T E R O N E , P R I N C I P A L A D R E N A L G L U C O C 0 R T I C O S T E R 0 ID IN T H E R A T HAS B E E N I N V E S T I G A T E D ( B U S H , 1 9 5 3 ) IN C O N T R O L L I N G T H E A C T I V I T Y OF T R Y P T O P H A N H Y D R O X Y L A S E IN R A T B R A I N . T H E T R Y P T O P H A N H Y D R O X Y L A S E A C T I V I T Y IN T H E R A T M I D - B R A I N D E C R E A S E S A F T E R A D R E N A L E C T O M Y AND IS R E S T O R E D B Y T R E A T M E N T W I T H C O R T I C O S T E R O N E ( E F R A I N AND M C E V E N , 1 9 6 9 ) . H Y D R O C O R T I S O N E , C O R T I S O N E AND ACTH H A V E B E E N R E P O R T E D TO D E C R E A S E E X T R A C E R E B R A L R A T T I S S U E 5 -H Y D R O X Y T R Y P T A M I N E (5HT) (HlCKS AND WE S T , 1 9 5 8 ; CA S S AND M A R S H A L L , 1 9 6 2 ) . A FEW S T U D I E S , W I T H A P P A R E N T L Y C O N F L I C T I N G R E S U L T S , H A V E B E E N M A D E , H O W E V E R , ON T H E E F F E C T O F A D R E N A L C O R T I C O I D S ON B R A I N 5HT. T H U S 5 0 MG C0R T I SON E / K G M / D A Y I N T R A P E R I T O N E A L L Y F O R T W E N T Y D A Y S WAS F O U N D TO I N C R E A S E R A T B R A I N 5H'f (G I A R M A N AND P E P E U , 1 9 6 2 ) , W H I L E 1 0 . I . U . ACTH / D A Y I N T R A M U S C U L A R L Y F O R T E N D A Y S C A U S E D A F A L L (R E S N I C K _E_T _AL . , 1 9 6 1 ) . |T HAS B E E N SHOWN T H A T H Y D R O C O R T I S O N E C A U S E S A F A L L IN B O T H 5HI A N D 5 -H Y D R O X Y I D O L A C E T I C A C I D (5H I A A) IN R A T B R A I N . LOW B R A I N 5HT IS NOT F O U N D A F T E R H Y D R O C O R T I S O N E I N J E C T I O N S A R E R E P E A T E D D A I L Y FOR 5 TO 1 3 D A Y S (CU R Z O N AND G R E E N , 1 9 6 8 ) . EXPERIMENTAL E X P E R I M E N T 1 TH I S E X P E R I M E N T WAS D E S I G N E D TO D E T E R M I N E W H E T H E R A D I E T A R Y D E F I C I E N C Y O F L Y S I N E ( .51$) A F F E C T S T H E P L A S M A C O N C E N T R A T I O N OF C O R T I C O S T E R O N E IN T H E C H I C K . THE C O M P O S I T I O N OF T H E B A S A L D I E T IS SHOWN IN TA B L E 1. A T O T A L OF 13$ O F P R O T E I N IN T H E D I E T WAS S U P P L I E D BY W H E A T AND I S O L A T E D S O Y B E A N P R O T E I N . T H E B A S A L D I E T ( D I E T 1) WAS C A L C U L A T E D TO C O N T A I N 0 .65$ O F L Y S I N E . D I E T 2 WAS S I M I L A R IN C O M P O S I T I O N TO D L E T 1 , B U T WAS S U P P L E M E N T E D W I T H 0 .15$ OF L Y S I N E IN T H E FORM OF L - L Y S I N E H Y D R O C H L O R I D E . EA C H D I E T WAS F E D TO 12 O N E - W E E K - O L D WH I T E LE G H O R N C O C K E R E L C H I C K S . PR I O R TO T H E E X P E R I M E N T A L P E R I O D T H E C H I C K S HAD B E E N F E D A C H I C K S T A R T I N G D I E T C O N T A I N I N G 20$ O F P R O T E I N . THE C H I C K S WERE K E P T IN T H E R M O S T A T I C A L L Y C O N T R O L L E D B A T T E R Y B R O O D E R S . T H E Y WERE G I V E N F E E D AND W A T E R AD L I B I T U M . L I G H T I N G WAS C O N T I N U O U S . P L A S M A C O R T I C O S T E R O N E L E V E L S WERE D E T E R M I N E D A F T E R T H E C H I C K S HAD B E E N ON E X P E R I M E N T F O R 14, 4 3 , AND 54 D A Y S . THE B L O O D WAS DRAWN F R O M T H E W I N G V E I N BY P U N C T U R I N G W I T H T H E T I P O F A S H A R P S C A L P E L I N T O HE PAR I N I Z E D T E S T T U B E S . T W E L V E C H I C K S F R O M E A C H G R O U P WERE B L E D . BL O O D WAS I M M E D I A T E L Y C E N T R I F U G E D F O R S E P A R A T I N G P L A S M A . C O R T I C O S T E R O N E C O N C E N T R A T I O N FROM P L A S M A WAS D E T E R M I N E D B Y F O L L O W I N G T H E F L U O R O M E T R I C M E T H O D OF G R O S S AND C O L M A N O , 1971. AC I D - W A S H E D C O N T A I N E R S WERE U S E D F O R W A T E R B L A N K S , C O R T I C O S T E R 0 N E S T A N D A R D (0 .25 U G M . / 0 . 5 M L . ) AND B L O O D P L A S M A (0 .50 M L . ) . P L A S M A OR S T A N D A R D WAS O V E R L A Y E R E D AND 1 8 .2,3 TABLE 1 . - - C O M P O S i T i O N O F E X P E R I M E N T A L D I E T I N G R E D I E N T S P E R C E N T SO F T WHEAT ( R O L L E D ) 84. 70 I S O L A T E D S O Y B E A N P R O T E I N 5. 60 BONE M E A L 2 . 00 L1 M E S T O N E 1 . 00 1OD1 Z E D S A L T 0 . 50 V E G E T A B L E O I L 2 . 00 CORN S T A R C H ^ M1CRONUTR1ENTS 3 . 70 0 . 50 TO T A L 100. 00 1 » M I C R O N U T R I E N T S : E A C H K I L O G R A M O F D I E T C O N T A I N E D M A N G A N E S E S U L P H A T E , 132 MG . ; R I B O F L A V I N , 3 .97 M G . j C A L C I U M P A N T O T H E N A T E , 9.26 M G . j F O L A C I N , 0.551 M G . ; P Y R I D O X I N E H Y D R O C H L O R I D E , 2 .866 MG . ; N I A C I N , 27 .05 M G . ; C H O L I N E C H L O R I D E , 1325 M G . ; V I T A M I N E, 18.22 M G . , V I T A M I N A, 4440 I.U.; V I T A M I N D 3, 440 I.U.; V I T A M I N B 1 ? , .0133 M G . ; ZN B A C I T R A C I N , 307.10 M G . ; A M P R O L 25$, 500.45 M G . 2 E X P E R I M E N T A L D I E T U S E D IN TR E A T M E N T 1, E X P E R i M E N T 1 . L Y S I N E WAS A D D E D A T 0 .15$ L E V E L TO E X P E R I M E N T A L D I E T IN TR I A L 2 T R E A T M E N T . S H A K E N W I T H 0.1 NORMAL S O D I U M H Y D R O X I D E ( 0 . 5 0 M L . ) , AND T H E N B R O U G H T TO 2.0 M L . V O L U M E W I T H D I S T I L L E D W A T E R . A L L D I L U T I O N S WERE O V E R -L A Y E R E D A N D W A S H E D BY S H A K I N G W I T H P E T R O L E U M E T H E R ( 2 . 0 M L . ) W H I C H WAS R E M O V E D . F R E S H L Y D I S T I L L E D ( 5 8 . 8 $ C . CUT-POINT) C H L O R O F O R M ( 3 . 0 M L . ) WAS A D D E D AND S H A K E N TO E X T R A C T T H E F L U O R O G E N S . A F T E R R E M O V A L O F SUPERNATANT L A Y E R , T H E C H L O R O F O R M WAS O V E R L A Y E R E D AND S H A K E N A G A I N W I T H 0.1 NORMAL S O D I U M H Y D R O X I D E . AN A L I Q U O T OF CHLOROFORM EXTRACT ( 2 . 0 M L . ) WAS T R A N S F E R R E D TO FLUOROMETER R E A D I N G T U B E S I N T O W H I C H F R E S H L Y P R E P A R E D 3 0 N S U L P H U R I C A C I D ( 4 . 8 M L . ) HAD B E E N A D D E D AND S H A K E N V I G O R O U S L Y . A F T E R 4 0 M I N U T E S A T ROOM T E M P E R A T U R E F L U O R E S C E N C E WAS R E C O R D E D W I T H A T U R N E R F L U O R O M E T E R 1 1 1 , U S I N G E X C I T A T I O N F I L T E R S 2A AND 4 7 B , AND E M I S S I O N F I L T E R S 5 8 AND 2 A 1 2 . WHEN T H E B I R D S HAD B E E N ON E X P E R I M E N T FOR 3 4 , 6 9 , AND 112 D A Y S OF A G E , T H E Y WERE S U B J E C T E D TO S T R E S S IN T H E FORM OF E L E C T R I C S H O C K A N D T H E L E V E L S OF P L A S M A C O R T I C O S T E R 0 N E M E A S U R E D A C C O R D I N G TO T H E P R O C E D U R E I N D I C A T E D A B O V E . THE E L E C T R I C S H O C K WAS A D M I N I S T E R E D T H R O U G H B A T T E R Y S H O C K E R , S I X B I R D S F R O M E A C H G R O U P WERE S U B J E C T E D TO 6, 9, AND 1 5 I N T E R M I T T E N T S H O C K S A T 3 4 , 6 9 , AND 1 1 2 D A Y S OF A G E R E S P E C T I V E L Y . E X P E R I M E N T 2 IN T H I S E X P E R I M E N T T H E E F F E C T S OF P R O T E I N D E F I C I E N C Y AND O F L Y S I N E D E F I C I E N C Y ( . 5 1 $ ) ON P L A S M A C 0 R T I C O S T E R O N E L E V E L WERE S T U D I E D , TWO B A S A L D I E T S C O N T A I N I N G 2 0 . 9 AND 1 3 . 0 $ O F P R O T E I N R E S P E C T I V E L Y WERE F O R M U L A T E D . E A C H OF T H E S E D I E T S WAS F E D W I T H AND W I T H O U T S U P P L E M E N T A R Y L Y S I N E . THE C O M P O S I T I O N O F T H E D I E T S IS SHOWN IN T A B L E 2 . IN O R D E R T H A T T H E 2 0 . 9 $ P R O T E I N B A S A L D I E T S H O U L D BE D E F I C I E N T IN L Y S I N E , S E S A M E S E E D M E A L WAS U S E D AS T H E S O U R C E OF S U P P L E M E N T A R Y P R O T E I N . THE 1 3 . 0 $ P R O T E I N B A S A L D I E T WAS F O R M U L A T E D TO BE D E F I C I E N T IN P R O T E I N B U T N O T IN L Y S I N E R E L A T I V E TO T H E A M O U N T O F P R O T E I N C O N T A I N E D IN T H E D I E T . I S O L A T E D S O Y B E A N P R O T E I N W A S , T H E R E F O R E , U S E D AS T H E S O U R C E OF S U P P L E M E N T A R Y P R O T E I N IN T H I S D I E T . TH E C H I C K S U S E D IN T H E E X P E R I M E N T WERE T H R E E - D A Y OLD WHITE L E G H O R N M A L E C H I C K S W H I C H HAD B E E N F E D A C H I C K S T A R T I N G . D I E T D U R I N G T H E T H R E E D A Y S A F T E R H A T C H I N G . T H E Y WERE D I S T R I B U T E D A T R A N D O M I N T O 16 L O T S OF 25 C H I C K S IN T H E C O M P A R T M E N T S OF 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 B R O O D E R S . E A C H E X P E R I M E N T A L D I E T WAS F E D TO Q U A D R U P L I C A T E L O T S O F C H I C K S . F E E D AND W A T E R WERE S U P P L I E D AD L I B I T U M . L I G H T I N G WAS C O N T I N U O U S . T H E C H I C K S WERE V A C C I N A T E D I N T R A O C U L A R L Y A G A I N S T N E W C A S T L E D I S E A S E A T T E N D A Y S O F A G E . TH E C H I C K S WERE W E I G H E D I N D I V I D U A L L Y E A C H W E E K . OX Y G E N C O N S U M P T I O N R A T E WAS M E A S U R E D IN 20 B I R D S F R O M E A C H O F T H E F O U R D I E T A R Y T R E A T M E N T S ON T H E 53RD D A Y O F T H E E X P E R I M E N T . P R I O R TO M E A S U R E M E N T O F O X Y G E N C O N S U M P T I O N T H E B I R D S WERE FASTTED F O R 24 HOURS B U T G I V E N W A T E R AD L I B I T U M . EACH B I R D WAS W E I G H E D AND T H E N P L A C E D IN T H E R E S P I R O M E T E R . THE R E S P I R O M E T E R C H A M B E R WAS F I L L E D W I T H TABLE 2. — C O M P O S I T I O N O F D I E T S IN E X P E R I M E N T 2. D 1 E T NUMBER I N G R E D I E N T S ( $ ) 1 2 2 4 WHEAT ( 1 2 . 5 $ P R O T E I N ) 6 9 . 0 0 6 9 .00 SO F T W H E A T ( 9 . 4 5 $ P R O T E I N ] ) - 84. 7 84. 7 V E G E T A B L E O I L 3. 0 0 3 . 0 0 2. 0 2. 0 BONE M E A L 1 . 0 0 1 .00 2. 0 2. 0 L 1 M E S T O N E 1 . 0 0 1 .00 1 . 0 1 . 0I O D I Z E D S A L T ^ M 1 C R O N U T R 1 E N T S 0. 5 0 0 .50 0. 5 0. 5 0. 5 0 0 .50 0. 5 0. 5 S E S A M E M E A L ( 5 0 . 6 9 $ P R O T E I N ) 2 4 . 2 0 24 .20 - -SO Y B E A N P R O T E I N ( 9 0 . 8 1 $ P R O T E I N ) - -- -- 5. 6 5. 6 L Y S I N E M O N O H Y D R O C H L O R I DE ( 9 8 $ ) 0 .765 0 1 9 1 3 CORN S T A R C H 80 .035 3. 7 3. 5 0 8 7 TO T A L 1 0 0 . 0 0 1 0 0 .00 1 0 0 . 0 0 1 0 0 . 0 0 C A L C U L A T E D A N A L Y S I S : CR U D E P R O T E I N ( $ ) 2 0 . 9 2 0 .9 1 3 . 0 0 1 3 . 0 0 L Y S I N E ( $ ) • 51 1 .11 • 6 5 • 80 ^ M 1 C R O N U T R 1 E N T S U S E D WERE T H E S A M E AS SHOWN 1 N E X P E R I M E N T 1 ( T A B L E 1 ) 2 3 O X Y G E N . OX Y G E N C O N S U M P T I O N WAS M E A S U R E D FOR S I X M I N U T E S . I N I T I A L AND F I N A L T E M P E R A T U R E S OF T H E C H A M B E R WERE N O T E D . B A R O M E T R I C P R E S S U R E WAS R E C O R D E D H O U R L Y . T H E C H A M B E R OF T H E R E S PI ROME T E R WAS C O V E R E D W I T H A L U M I N U M F O I L TO P R O V I D E A D A R K E N V I R O N M E N T S O T H A T O X Y G E N C O N S U M P T I O N WOULD BE M E A S U R E D W H I L E T H E B I R D S WERE IN A R E S T I N G S T A T E . R E S P I R A T I O N WAS M E A S U R E D IN B I R D S F R O M T H E D I F F E R E N T T R E A T M E N T S IN R O T A T I O N TO A V O I D T H E I N F L U E N C E OF D I U R N A L F L U C T U A T I O N ON T H E R E S U L T S O B T A I N E D W I T H T H E E X P E R I M E N T A L T R E A T M E N T S . T H E M E A S U R E M E N T S WERE M A D E O V E R A P E R I O D FROM 9 A : M : TO 1 A : M : T H E F O L L O W I N G M O R N I N G . OX Y G E N C O N S U M P T I O N R A T E WAS C A L C U L A T E D AS M L . P E R K G M . B O D Y W E I G H T P E R S E C O N D B Y T H E F O L L O W I N G F O R M U L A (L OWE L L _ET _AL . , 1 9 5 9 ) . O B S . M L . x 2 7 3 ° A x ( B . P . - V.P.) x 1QOO  £>BS. A V E . T E M P . C ° + 2 7 3 ° ) x 7 6 0 M . M . x B.W. (G M S) x T I M E ( S E C ) I N I T I A L + F I N A L O B S . A V E . T E M P . = ^ BPP. = BA R O M E T R I C P R E S S U R E V.P. = VA P O U R P R E S S U R E A = A B S O L U T E AD R E N A L G L A N D S WERE W E I G H E D FROM 2 0 O F T H E B I R D S F E D E A C H O F T H E E X P E R I M E N T A L D I E T S F O R 4 5 D A Y S . B O D Y W E I G H T S WERE M E A S U R E D B E F O R E T H E B I R D S WERE K I L L E D FOR R E M O V A L O F T H E A D R E N A L G L A N D S . P L A S M A C O R T I C O S T E R O N E L E V E L S WERE D E T E R M I N E D A F T E R T H E C H I C K S HAD B E E N ON E X P E R I M E N T F O R 4 5 A N D5§4 D A Y S . T W E L V E C H I C K S F R O M E A C H T R E A T M E N T WERE B L E D A T 4 5 D A Y S O F A G E . AT 54 D A Y S , F O U R T E E N C H I C K S WERE U S E D T H O S E WERE U N D E R T H E T R E A T M E N T OF 2 0 . 9 $ P R O T E I N W I T H AND W I T H O U T L Y S I N E S U P P L E M E N T A T I O N . T H E L E V E L S O F P L A S M A C 0 R T I C O S T E R 0 N E WERE M E A S U R E D A C C O R D I N G TO T H E P R O C E D U R E • I ND I C A T E D IN E X P E R I M E N T 1. 24 :NT EXPERIMENT 3 THE DESIGN OF THE EXPERIMENT WAS S I M I L A R TO THAT OF EXPERIMEI 2 . THE BASAL DIETS CONTAINED 2 0 . 9 $ OR 13.2$ or PROTEIN. |N EXPERIMENT 3 , HOWEVER, THE 13.2$ PROTEIN DIET CONTAINED SOYBEAN MEAL IN PLACE OF ISOLATED SOYBEAN PROTEIN AS THE SOURCE OF SUPPLEMENTARY PROTEIN. THE COMPOSITION OF THE DIETS IS SHOWN IN TABLE 3 . DAY-OLD WHITE LEGHORN MALE CHICKS WERE DIS T R I B U T E D AT RANDOM INTO 12 LOTS OF 12 CHICKS IN THE COMPARTMENTS OF E L E C T R I C A L L Y HEATED BATTERY BROODERS. EACH OF THE EXPERIMENTAL DIETS WAS FED TO T R I P L I C A T E LOTS OF CHICKS. THE CHICKS WERE GIVEN FEED AND WATER AD L I B I T U M . L I G H T I N G WAS CONTINUOUS. THE CHICKS WERE VACCINATED INTRAOCULARLY AGAINST NEWCASTLE DISEASE WHEN THEY WERE TEN DAYS OLD. A T WEEKLY INTERVALS THE CHICKS WERE I N D I V I D U A L L Y WEIGHED. WHEN CHICKS WERE 2, 4 , AND 6 WEEKS OF AGE BLOOD WAS COLLECTED BY HEART PUNCTURE FROM S I X OF THE BIRDS FED EACH D I E T . PLASMA CORTICOSTERONE CONCENTRATIONS WERE DETERMINED ACCORDING TO THE METHOD ALREAD Y DESCRI BED. THE BODY WEIGHT WAS RECORDED FOR EACH BIRD BEFORE COLLECTION OF BLOOD. THE PLASMA CORTICOSTERONE DETERMINED FOR EACH BIRD WAS GRAPHED AGAINST BODY WEIGHT AT 2 , A, AND 6 WEEKS OF AGE. 25 TABLE 3 . - - C O M P O S i T I O N OF D I E T S IN E X P E R I M E N T 3 . 1 I N G R E D I E N T S ( $ ) WHEAT V E G E T A B L E O I L BONE M E A L L i M E S T O N E I O D I Z E D S A L T S E S A M E M E A L (50.69$ P R O T E I N ) SO Y B E A N M E A L (48.5$ P R O T E I N ) M I C R O N U T R I E N T s ' ' L Y S I N E M O N O H Y D R O C H L O R I D E CORN S T A R C H TO T A L D I E T NU M B E R 1 2 3 4 6 8 . 5 6 8 . 5 84.2 84.2 3 . 0 3 . 0 2 . 0 2 . 0 2 . 0 2 . 0 2 . 5 2 . 5 0 . 5 0 . 5 1 . 0 1 . 0 0 . 5 0 . 5 0 . 5 0 . 5 24 .2 24 .2 5 .6 5 .6 0 . 5 0 . 5 0 . 5 0 . 5 0.765 0.3061 0 . 8 0.035 3 . 7 3.3939 100.00 100. 1 00 . 100. C A L C U L A T E D A N A L Y S I S : CR U D E P R O T E I N ($) 2 0 . 9 2 0 . 9 13.2 13.2 0 . 4 5 0 .69 L Y S I N E ($) 0.51 1.11 M l C R O N U T R I E N T S USED WERE THE SAME AS SHOWN IN E X P E R I M E N T 1 ( T A B L E 1 ) . 26 E X P E RI M E N T 4 E X P E R I M E N T 4 W A S C O N D U C T E D TO D E T E R M I N E W H E T H E R D I E T A R Y F A C T O R S , O T H E R T H A N P R O T E I N L E V E L O R A M I N O A C I D D E F I C I E N C Y W H I C H A F F E C T M E T A B O L I C R A T E , A L T E R P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S . THE E F F E C T S O F E X C E S S V I T A M I N E A N D T H E G O I T R O G E N T H I O U R A C I L W E R E S T U D I E D . THE C O M P O S I T I O N O F T H E B A S A L D I E T E M P L O Y E D A N D T H E D I E T A R Y T R E A T M E N T S A R E G I V E N I N T A B L E 4. D A Y - O L D W H I T E L E G H O R N M A L E C H I C K S W E R E R A N D O M L Y D I S T R I B U T E D I N T O E I G H T L O T S O F 25 C H I C K S E A C H I N T H E C O M P A R T M E N T S O F E L E C T R I C A L L Y H E A T E D B R O O D E R S . T H E B A S A L D I E T A N D T H E D I E T C O N T A I N I N G A N E X C E S S O F V I T A M I N E W E R E E A C H F E D T O F O U R R E P L I C A T E L O T S O F C H I C K S . A T O N E - W E E K O F A G E T H E C H I C K S W E R E V A C C I N A T E D I N T R A O C U L A R L Y A G A I N S T N E W C A S T L E D I S E A S E . A T 40 D A Y S O F A G E 48 C H I C K S F R O M E A C H D I E T W E R E T R A N S F E R R E D T O C A G E S I N T W O T E M P E R A T U R E - C O N T R O L L E D C H A M B E R S H E L D A T 13°C A N D 2 4 °C R E S P E C T I V E L Y . T H E R E W E R E 12 C A G E S I N E A C H C H A M B E R A N D F O U R C H I C K S W E R E P L A C E D I N E A C H . E A C H C H A M B E R A C C O R D I N G L Y H O U S E D 24 C O N T R O L C H I C K S , A N D 24 C H I C K S W H I C H H A D B E E N F E D T H E E X C E S S O F V I T A M I N E. A F U R T H E R T R E A T M E N T W A S I M P O S E D A T T H I S T I M E . T H I O U R A C I L A T A L E V E L O F 0.1$ W A S A D D E D T O T H E D I E T O F 24 O F T H E C H I C K S F E D E A C H D I E T I N E A C H C H A M B E R . F E E D A N D W A T E R W E R E S U P P L I E D A D L I B I T U M D U R I N G T H E E X P E R I M E N T A N D L I G H T I N G W A S C O N T I N U O U S . B L O O D S A M P L E S W E R E T A K E N F R O M T H E W I N G V E I N O F T H R E E B I R D S F R O M E A C H E X P E R I M E N T A L T R E A T M E N T . P L A S M A C 0 R T I C O S T E R O N E C O N C E N T R A T I O N S W E R E D E T E R M I N E D A C C O R D I N G T O T H E M E T H O D A L R E A D Y D E S C R I B E D . TABLE 4 . - - C O M P O S i T I O N OF D I E T AND E X P E R I M E N T A L OU T L I N E IN E X P E R I M E N T 4 . I N G R E D I E N T S P E R C E N T WHEAT H E R R I N G M E A L BONE M E A L L i M E S T O N E D I S T I L L E R ' S D R I E D S O L U B L E S I O D I Z E D S A L T TO T A L M I C R O N U T R I E N T S : EA C H K G . OF D I E T WAS S U P P L E M E N T E D W I T H : M A N G A N E S E S U L P H A T E , 150 M G . ; V I T A M I N A, 5500 I.U.; V I T A M I N D 3, 440 I.U.; M E N A D I O N E , 0 . 6 0 M G . ; R I B O F L A V I N , 3 . 6 M G . ; C A L C I U M P A N T O T H E N A T E , 10 M G . ; F O L A C I N , 1.2 M G . ; C H O L I N E C H L O R I D E , 1300 M G . ; A M P R O L - 2 5 , 500 M G . E X P E R I M E N T A L O U T L I N E : TR E A T M E N T 1 - D I E T 1 TR E A T M E N T 2 = D I E T 1 + 0 .1$ T H I O U R A C I L T R E A T M E N T 3 = D I E T 1 + V I T A M I N E ( V I T A M I N E 110 I.U. P E R K G . UP TO 2 W E E K S ; L A T E R 220 I.U. P E R KG . ) TR E A T M E N T 4 = D I E T 1 + V I T A M I N E (220 I.U. P E R K G . ) + T H I O U R A C I L , 0 . 1 $ . 81 . 5 14.5 0 . 5 1 . 0 2 . 0 0 . 5 100.0 28 EXPERIMENT 5 ON THE BASIS OF THE ABNORMAL BEHAVIOUR EXHIBITED BY BIRDS IN EXPERIMENT 2 FED THE LYSINE DEFICIENT DIET CONTAINING 2 0 . 9 % OF PROTEIN, AN EXPERIMENT WAS CONDUCTED SPECIF ICALLY TO INVESTIGATE THE PHENOMENON. DAY-OLD WHITE LEGHORN CHICKS OF DEKALB STRAIN WERE RANDOMLY DISTRIBUTED INTO EIGHT LOTS OF 2 0 CHICKS EACH IN THE COMPARTMENTS OF ELECTRICALLY HEATED BATTERY BROODERS. THE EXPERIMENTAL DIETS EMPLOYED WERE OF THE SAME COMPOSITION AS DIETS 1 AND 2 IN EXPERIMENT 2 (TABLE 2 ) . EACH DIET WAS FED TO QUADRUPLICATE LOTS OF CHICKS. FEED AND WATER WERE PROVI PEP AD LI BI TUM. LIGHTING WAS CONTINUOUS. THE CHICKS WERE WING-BANDED AND VACCINATED INTRAOCULARLY AGAINST NEWCASTLE DISEASE AT ONE WEEK OF AGE. INDIVIDUAL BODY WEIGHTS WERE RECORDED WEEKLY. A T 4 0 DAYS OF AGE 1 0 CHICKS FROM EACH TREATMENT WERE KILLED. THE BRAIN WAS DISSECTED FROM EACH CHICK AND THE BRAIN STEM AND THE CEREBRAL HEMISPHERES WERE WEIGHED. BY THE TIME THE BIRDS WERE 45 DAYS OLD THOSE FED THE LYSINE-DEFICIENT DIET WERE EXTREMELY WEAK. |N ORDER TO MAINTAIN THE POPULATION OF BIRDS FROM THIS DIETARY TREATMENT THE BIRDS WERE FED THE LYSINE-SUPPLEMENTED DIET FOR FOUR WEEKS. AT THE END OF THIS TIME THE BIRDS WERE RE-FED THE LYSINE DEFICIENT DIET . TEN DAYS LATER, WHEN THEY WERE 83 DAYS OLD, FOUR BIRDS (1 MALE AND 3 FEMALES) WERE TESTED FOR EXPLORATORY BEHAVIOUR IN SEARCH OF FOOD IN SIMULTANEOUS TRIALS FOR TWO GROUPS OF BIRDS. THE FOLLOWING TEST PROCEDURE WAS DEVISED. THE BIRDS WERE PLACED IN PORTABLE INDIVIDUAL CAGES AND FEED WAS W I T H D R A W N F R O M 5 P . M . U N T I L T H E B E H A V I O U R T R I A L WAS C O N D U C T E D T H E F O L L O W I N G M O R N I N G A T 10 A . M . T H E D E S I G N OF T H E S I M P L E M A Z E IS SHOWN IN F I G U R E 1 . TH E C A G E WAS P O S I T I O N E D A T T H E E N T R A N C E TO T H E F E E D A T A D I S T A N C E OF 15 F E E T AND T H E C A G E DOOR O P E N E D . S U C C E S S F U L E X P L O R A T I O N T I M E WAS T H E T I M E E L A P S I N G B E T W E E N O P E N I N G OF T H E C A G E DOOR AND D I S C O V E R Y OF T H E F E E D BY A N Y M E M B E R O F T H E G R O U P . T H E P R O C E D U R E WAS R E P E A T E D FOR F I V E D A Y S . T H E L O C A T I O N O F F E E D WAS T H E N C H A N G E D AND E X P L O R A T I O N T I M E WAS N O T E D F O R T H R E E D A Y S M O R E . A S E C O N D T E S T OF E X P L O R A T O R Y B E H A V I O U R U S E D A 5 X 5 G R I D O F 9 - I N C H S Q U A R E S R U L E D ON T H E F L O O R ( F I G . 2 ) . T H E G R I D WAS C O V E R E D B Y A Y S C R E E N E N C L O S U R E 30 I N C H E S H I G H . T H E ROOM ; WAS D A R K AND T H E G R I D WAS I L L U M I N A T E D BY A 60 W A T T I N C A N D E S C E N T L I G H T B U L B S U S P E N D E D A T T H E T O P . A T 83 D A Y S O F A G E I N D I V I D U A L B I R D S WERE S U B J E C T E D TO E X P L O R A T O R Y T E S T . EA C H C H I C K WAS P L A C E D IN T H E S A M E S Q U A R E I N S I D E T H E G R I D AND M O V E M E N T S WERE O B S E R V E D F O R T H R E E M I N U T E P E R I O D S . E L E V E N B I R D S WERE S U B J E C T E D TO T H E T E S T C O N D U C T E D IN T H E A B O V E WAY AND NONE SHOWED A N Y M O V E M E N T , I T WAS T H O U G H T T H A T T H E B I R D S M I G H T NOT BE M O V I N G B E C A U S E T H E Y WERE F R I G H T E N E D A T B E I N G I S O L A T E D F R O M O T H E R B I R D S . A T A P E R E C O R D I N G O F T H E S O U N D S M A D E B Y A F L O C K OF L A Y I N G B I R D S WAS T H E R E F O R E P L A Y E D FOR T H R E E M I N U T E S A F T E R E A C H B I R D WAS P L A C E D ON T H E G R I D . T H E S E S O U N D S E V I D E N T L Y WERE R E A S S U R I N G TO T H E B I R D S AND T H E Y SHOWED A P P A R E N T L Y N O R M A L M O V E M E N T S W I T H T H I S B A C K G R O U N D N O I S E . T H I S H E L P E D T H E B I R D S TO S T A R T M O V E M E N T S . E L E V E N C H I C K S F R O M E A C H E X P E R I M E N T A L T R E A T M E N T WERE S U B J E C T E D TO T H E E X P L O R A T O R Y T E S T E A C H D A Y F O R F I V E D A Y S . 30 F I G U R E 2 MA Z E TOR S T U D Y OF E X P L O R A T O R Y B E H A V I O U R IN E X P E R I M E N T S 5. RESULTS E X P E R I M E N T 1 P L A S M A C O N C E N T R A T I O N S OF C O R T I C O S T E R O N E , D E T E R M I N E D WHEN C H I C K S WERE 2 1 , 50 , AND 61 D A Y S O F A G E , A R E SHOWN IN T A B L E 5 . T H E A N A L Y S I S O F V A R I A N C E O F T H E D A T A IS P R E S E N T E D IN A P P E N D I X T A B L E I. P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N WAS H I G H E R A T 21 AND 50 D A Y S O F A G E IN T H E C H I C K S 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 T H A N IN T H E C H I C K S F E D T H E D I E T S U P P L E M E N T E D W I T H L Y S I N E . T H E D I F F E R E N C E WAS S I G N I F I C A N T , H O W E V E R , O N L Y IN T H E M E A S U R E M E N T S M A D E ON T H E 50TH D A Y . A T 61 D A Y S O F A G E T H E R E WAS NO S I G N I F I C A N T D I F F E R E N C E IN T H E A V E R A G E P L A S M A C O N C E N T R A T I O N O F C O R T I C O S T E R O N E B E T W E E N T H E B I R D S R E C E I V I N G T H E TWO D I E T A R Y L E V E L S O F L Y S I N E . T H E E F F E C T S ON P L A S M A C O N C E N T R A T I O N OF C 0 R T I C O S T E R O N E O F S U B J E C T I N G T H E C H I C K S F E D T H E D I F F E R E N T L E V E L S O F L Y S I N E TO A M I L D E L E C T R I C S H O C K A R E SHOWN IN T A B L E 2 AND F I G U R E 3 . T H E N U M B E R OF B I R D S T E S T E D WAS S M A L L AND T H E R E WAS C O N S I D E R A B L E V A R I A T I O N ON T H E C O R T I C O S T E R O N E V A L U E S F O R B I R D S R E C E I V I N G T H E S A M E T R E A T M E N T . NO C O N S I S T E N T P A T T E R N OF E F F E C T O F S H O C K T R E A T M E N T ON C O R T I C O S T E R O N E L E V E L S WAS O B S E R V E D . 32 TABLE 5 . - - P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN CH I C K S F E D A 13$ P R O T E I N D I E T W ITH AND W I T H O U T S U P P L E M E N T A R Y L Y S I N E IN E X P E R I M E N T 1. L Y S I N E D E F I C I E N T L Y S I N E S U P P L E M E N T AG E UGMS/100 M L . ± S.D. UGMS/100 M L . ± S.D. P R O B A B I L I T Y 21 D A Y S 4 6 . 6 ± 7 . 3 (11) 38 .6 ± 13 .0 (10) > . 0 5 50 4 7 . 0 ± 7 . 0 (11) 38 .4 ± 3 . 6 (10) «=.01 61 5 6 . 9 ± 15 .3 (10) 6 1 . 5 ± 16 .9 ( 9) > . 0 5 NOT S T R E S S E D S T R E S S E D  AG E UGMS/100 M L . ± S.D. UGMS/100 M L . ± S.D. P R O B A B I L I T Y L Y S i N E D E F I C I E N T  34 38 .10 ± 5 . 3 (3) 34 .2 ± 4 . 6 0 (6) > .06 69 59 .7 ± 12.76 (3) 55 .4 ± 10.98 (6) > . 0 5 112 83.1 ± 13.24 (3) 5 6 . 8 ± 15.21 (6) > .05 L Y S I N E S U P P L E M E N T  34 3 5 . 5 ± 2 . 4 5 (3) 3 3 . 3 ± 2 .77 (6) > . 0 5 69 42 .2 ± 8 .49 (4) 57.1 ± 8 .03 (6) < . 0 5 1 12 7 8 . 8 ± 3 . 8 5 (3) 6 8 . 0 ± 8 .45 (5) > .05 F I G U R E S IN P A R E N T H E S E S SHOW T H E N U M B E R OF D E T E R M I N A T I O N S . 34 AGE IN DAYS VARIATION WITH THE AGE IN PLASMA CORTICOSTERONE CONCENTRATIONS IN CHICKS FED A 13$ PROTEIN DIET WITH AND WITHOUT SUPPLEMENTARY LYS I NE IN EXPERIMENT 1 . E X P E R I M E N T 2 P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN C H I C K S F E D T H E D I F F E R E N T D I E T S IN E X P E R I M E N T 2 A R E SHOWN IN T H E T A B L E 6. THE S T A T I S T I C A L A N A L Y S I S OF D A T A IS P R E S E N T E D IN T H E A P P E N D I X T A B L E 3. D U N C A N ' S NEW M U L T I P L E R A N G E T E S T ( D U N C A N , 1 9 5 5 ) SHOWS S I G N I F I C A N T D I F F E R E N C E IN T H E R E S P O N S E S TO T H E D I F F E R E N T D I E T S . H L G H P R O T E I N ( 2 0 . 9 % ) A S S O C I A T E D W I T H LOW L Y S I N E ( 0 . 5 1 % ) I N C R E A S E D T H E P L A S M A C O R T I C O S T E R 0 N E C O N C E N T R A T I O N S I G N I F I C A N T L Y AS C O M P A R E D TO 2 0 . 9 % P R O T E I N W I T H 1 . 1 1 % L Y S I N E OR 1 3 % P R O T E I N W I T H 0 . 6 5 % L Y S I N E . T H E T R E A T M E N T W I T H 1 3 % P R O T E I N AND 0 . 8 0 % L Y S I N E R E S U L T E D IN AN I N T E R M E D I A T E P L A S M A C O N C E N -T R A T I O N O F C O R T I C O S T E R O N E W H I C H WAS NOT 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 E C O N C E N T R A T I O N OF T H E O T H E R T R E A T M E N T S . T H E S E R E S U L T S WERE O B S E R V E D A F T E R T H E C H I C K S HAD B E E N ON T H E E X P E R I M E N T A L D I E T S FOR 4 5 D A Y S . T H E P L A S M A C O R T I C O S T E R O N E WAS M E A S U R E D IN C H I C K S U N D E R T R E A T M E N T O F 2 0 . 9 % P R O T E I N IN A S S O C I A T I O N W I T H 0 . 5 1 % L Y S I N E AND 1 . 1 1 % L Y S I N E R E S P E C T I V E L Y A F T E R T H E C H I C K S HAD B E E N ON E X P E R I M E N T A L D I E T S FOR 54 D A Y S . S T A T I S T I C A L A N A L Y S I S OF D A T A ( A P P E N D I X TA B L E 3 ) S H O W E D T H A T V A L U E S D E T E R M I N E D A T T H I S T I M E WERE NOT S I G N I F I C A N T L Y D I F F E R E N T , T H O U G H T H E C O R T I C O S T E R 0 N E L E V E L WAS H I G H E R W I T H LOW L Y S I N E T R E A T M E N T . T H E R E S P I R A T I O N R A T E S O F T H E C H I C K S F E D T H E D I F F E R E N T D I E T S IN E X P E R I M E N T 2 A R E SHOWN IN TA B L E 7. D U N C A N ' S NEW M U L T I P L E R A N G E T E S T ( D U N C A N , 1 9 5 5 ) SHOWED T H A T O X Y G E N C O N S U M P T I O N OF B I R D S F E D 2 0 . 9 % P R O T E I N W I T H 0 . 5 1 % L Y S I N E WAS S I G N I F I C A N T L Y G R E A T E R T H A N T H A T OF B I R D S F E D 2 0 . 9 % P R O T E I N AND 1 . 1 1 % L Y S I N E . O X Y G E N C O N S U M P T I O N R A T E S OF T H E C H I C K S F E D LOW P R O T E I N ( 1 3 . 0 % ) D I E T S W I T H 0 . 6 5 % AND 0 . 8 0 % L Y S I N E WERE B E T W E E N T H E V A L U E S O B T A I N E D W I T H H I G H P R O T E I N ( 2 0 . 9 % ) , TABLE 6 . — P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN CH I C K S FE D D I F F E R E N T D I E T S IN E X P E R I M E N T 2 . D I E T A R Y TR E A T M E N T S DAYS OF 1 2 A G E ' ^ NUMBER CR U D E P R O T E I N L Y S I N E Wo) 45 UGMS/100 M L . ± S . D . 54 UGMS/100 M L.±S .D. 1 2 0 . 9 0.51 7 4 . 4 1 A ± 25.56 (12) 5 9 . 0 6 A ± 2 0 . 3 5 (14) 2 2 0 . 9 1.11 4 3 . 5 8 B ± 15.13 (12) 4 6 . 8 9 A ± 23 .23 (13) 3 13 .0 0 . 6 5 5 2 . 6 4 B ± 15.99 (12) 4 1 3 . 0 0 .80 5 8 . 5 6 A B ± 17.88 (12) 1 -F i G U R E S IN P A R E N T H E S E S SHOW T H E N U M B E R O F D E T E R M I N A T I O N S . TR E A T M E N T S W I T H D I F F E R E N T S U P E R S C R I P T S A R E D I F F E R E N T (P < 0 . 0 1 ) . TABLE 7 . — R E S P I R A T I ON RA T E OF CH I C K S FE D D I F F E R E N T D I E T S IN E X P E R I M E N T 2. D I E T A R Y TR E A T M E N T S OX Y G E N CO N S U M P T I O N NUMBER P R O T E I N (*) L Y S 1 N E (*) B.W. ( G M ) ML ML 0? , KG B.W./ S E C . S.D. ML Op/HR B.W. (G M ) B.W.' 7 5 1 2 0 . 9 0.51 1 5 0 . 0 5 7 3 A ± .0241 ( 2 0 ) .206 .722 2 2 0 . 9 1.11 4 7 7 . 0 3 1 7 B ± .0098 ( 2 0 ) .114 .534 3 1 3 . 0 0.65 2 6 6 . 0 4 2 1 A B ± . 0 2 7 2 ( 2 0 ) .152 .612 4 ' 1 3 . 0 0.80 2 8 4 . 0 4 4 8 A B ± . 0 2 1 5 ( 2 0 ) .161 .662 1 F I G U R E S IN P A R E N T H E S E S SHOW T H E N U M B E R O F D E T E R M I N A T I O N S . 2 V A L U E S W I T H D I F F E R E N T S U P E R S C R I P T S A R E D I F F E R E N T (P «= 0.01.) H O W E V E R , T H E S E V A L U E S (0 .65 AND 0 .8$ L Y S I N E ) WERE NOT 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 0.51 AND 1.11 P E R C E N T D I E T A R Y L Y S I N E . AD R E N A L W E I G H T S IN MG P E R 100 GMS B O D Y W E I G H T O F C H I C K S A R E P R E S E N T E D IN T A B L E 8 . D U N C A N ' S NEW M U L T I P L E R A N G E T E S T ( D U N C A N , 1955) ON T H E M E A N S O F D I F F E R E N T T R E A T M E N T S SHOWED T H A T C H I C K S F E D 2 0 . 9 $ P R O T E I N IN A S S O C I A T I O N W I T H 0 .51$ L Y S I N E HAD S I G N I F I C A N T L Y H E A V I E R A D R E N A L W E I G H T S P E R 100 G R A M S O F B O D Y W E I G H T T H A N D I D C H I C K S F E D 20 .9$ P R O T E I N W I T H 1.11$ L Y S I N E OR 13.0$ P R O T E I N W I T H E I T H E R 0.65 OR 0 .80 P E R C E N T D I E T A R Y L Y S I N E . C H I C K S R E C E I V I N G 20 .9$ P R O T E I N W I T H 1.11$ L Y S I N E HAD T H E L O W E S T A D R E N A L W E I G H T S R E L A T I V E TO B O D Y W E I G H T S B U T T H E A D R E N A L W E I G H T S WERE NOT S I G N I F I C A N T L Y D I F F E R E N T FROM C H I C K S F E D 13.0$ P R O T E I N AND 0 .80$ L Y S I N E , A L T H O U G H T H E Y WERE D I F F E R E N T F R O M T H O S E OF C H I C K S F E D 13.0$ P R O T E I N AND 0 .65$ L Y S I N E . GROWTH R A T E S OF C H I C K S F E D D I F F E R E N T D I E T S A R E SHOWN IN F I G U R E 4 . B I R D S F E D ON R E Q U I R E D A M O U N T OF P R O T E I N (20.9$) AND L Y S I N E (1.11$) HAD T H E B E S T G R O W T H AND WERE T H E H E A V I E S T AMONG T H E V A R I O U S T R E A T M E N T S U S E D IN E X P E R I M E N T 2 . T H E R E L A T I O N S H I P B E T W E E N C 0 R T I C O S T E R O N E C O N C E N T R A T I O N S AND B O D Y W E I G H T S IS SHOWN IN F I G U R E S 5 , 6, AND 7. C O R T I C O S T E R O N E L E V E L D I D NOT SHOW A N Y R E L A T I O N S H I P W I T H R E S P E C T TO W E I G H T S A T 2 , 4 , AND 6 W E E K S O F A G E F O R E A C H T R E A T M E N T U S E D . TABLE 8.--AD R E N A L WE I G H T S OF CH I C K S FE D D I F F E R E N T D I E T S IN E X P E R I M E N T 2. D I E T A R Y TR E A T M E N T S L B.W. AD R E N A L WTS . AD R E N A L WT S . ( M G ) P R O T E I N {%) L Y S I N E {%) GMS ± S.D. MG ± S.D. 1 0 0 GMS B.W. 2 0 . 9 0.51 1 5 0 + 1 8 3 5 ± 8 2 2 . 1 7 A ± 3.1 3 ( 2 0 ) 2 0 . 9 1.11 4 7 7 + 5 6 64 ± 11 13 . 5 2 ° ± 2.75 ( 2 0 ) 1 3 . 0 0.65 2 6 6 + 5 9 3 9 ± 6 1 5 . 2 1 B ± 2.95 ( 2 0 ) 1 3 . 0 0.80 2 8 4 + 6 3 41 ± 8 • 1 4 . 6 4 B C ± 1 .89 ( 2 0 ) F I G U R E S IN P A R E N T H E S E S SHOW T H E N U M B E R O F D E T E R M I N A T I O N S . V A L U E S W I T H D I F F E R E N T S U P E R S C R I P T 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 <= 0.01 ). 4 0 FIGURE- 4 GROWTH RATES OF CHICKS FED DIFFERENT DIETS IN EXPERIMENT 2 . 'F IGURE 5 0 O R-1 R-2 R-3 R-4 PR O T E I N 20,9$ 20.9$ 13.2# 13.2$ L Y S I N E * 1% 1.1 0 © ,8 X o o 70 BODY WE TGHTS I N, G R A M S REl.A i ; ONSh ; P BETWEEN BODY WE i G H T S AND CORT I COSTERONE C C\ C E N T R A T I ON AT TWO WEEKS OF AGE IN CHICKS I ED D I ; F t. K E N r D I E T S . FIGURE 6 oI3d 5W PROTEIN LYSINE R- 1 20 .9$ .51$ e • R- 2 20.9% 1.11$ X X R- 3 13.2$ .45$ o o R- 4 13.2$ .69i 0 tg) o 0 o L 9o 12.0 i ISO 180 210 2*f0 BODY WE I G H T S IN G R A M S 300 330 RELATIONSHIP BETWEEN BODY WEIGHTS AND CORTICOSTERONE CONCENTRATION AT TOUR WEEKS OF AGE IN CLICKS FED - : r r t : « ^ r D i E T S . -p. ro FIGURE 7 PROTEIN LYSINE R-1 20.9$ .51$ 0 R-2 20. 9i 1 .11$ X X R-3 13.2$ .45$ O O R -4 ,69fo ® ® o l 5 Q O aw 250 i 1—! ~1 300 352? 4=00 Boo Y wr • r . H i s ; N GMS . 550 RELATIONSHIP BETWEEN BODY WEIGHTS AND CORTICOSTERONE CONCENTRAT:OM AT S ;X WEEKS o r AGE IN CH;CKS FED DIFFERENT ': I E TS . -J> co E X P E R I M E N T 3 ' P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN C H I C K S F E D D I F F E R E N T D I E T S IN E X P E R I M E N T 3 A R E SHOWN IN TA B L E 9 . S T A T I S T I C A L A N A L Y S I S O F D A T A F R O M T A B L E 9 F O R T H R E E D I F F E R E N T A G E S IS P R E S E N T E D IN A P P E N D I X TA B L E 4 . AT TWO W E E K S OF A G E C O R T I C O S T E R O N E C O N C E N T R A T I O N IN T H E P L A S M A OF T H E C H I C K S F E D D L E T 1 (20.9$ P R O T E I N W I T H 0 .51$ L Y S I N E ) WAS S I G N I F I C A N T L Y H I G H E R T H A N T H E C O N C E N T R A T I O N IN T H E P L A S M A OF T H E C H I C K S F E D T H E O T H E R T H R E E D I E T S (20.9$ P R O T E I N W I T H 1.11$ L Y S I N E , AND 13$ P R O T E I N W I T H .45$ AND .69$ L Y S I N E ) . T H E D I F F E R E N C E IN C O R T I C O S T E R O N E C O N C E N T R A T I O N WAS A L S O E V I D E N T A T 4 AND 6 W E E K S O F A G E , H O W E V E R , T H E S E O B S E R V A T I O N S WERE NOT S T A T I S T I C A L L Y S I G N I F I C A N T . GROWTH R A T E S OF C H I C K S F E D D I F F E R E N T D I E T S A R E SHOWN IN F I G U R E 8. R E Q U I R E D A M O U N T OF P R O T E I N (20.9$) A S S O C I A T E D W I T H B A L A N C E D L Y S I N E (1 .11$) AND O T H E R A M I N O A C I D S HAD T H E B E S T E F F E C T ON G R O W T H . L I G H T E S T , B I R D S WERE O B T A I N E D W I T H R E Q U I R E D P R O T E I N (20.9$) B U T LOW L Y S I N E ( . 5 1 $ ) . F U R T H E R M O R E , R E S U L T S I N D I C A T E T H A T T H E L Y S I N E R E Q U I R E M E N T FOR G R O W T H A T LOW L E V E L S OF P R O T E I N IS IN A P P R O X I M A T E L Y T H E S A M E R A T I O AS T H A T OF P R O T E I N AND L Y S I N E F O R O P T I M U M G R O W T H . TABLE 9 .--BO D Y WE I G H T AND P L A S M A C O R T I C O S T E R O N E CO N C E N T R A T I O N S IN CH I C K S FE D D I F F E R E N T D I E T S IN E X P E R I M E N T 3 . Two-1 2 WEEKS ' 1 2 FO U R -W E E K S ' 1 2 S I X - W E E K S ' P E R C E N T C O R T I C O S T E R O N E G O R T 1 C O S T E R O N E S O R T i C O S T E R O N E P R O T E I N L Y S i N E B.W. : t S .D. UGM/100 M L S B.W, 3M ) UGM/100 M L S B.W. • ( G M ) UGM/100 M L S 1) 2 0 . 9 0.51 72 + 6 6 2 . 1 8 A ± 7.82 124 + 1 8 9 1 . 9 8 A ± 31 .97 206 + 27 6 9 . 2 3 A ± 13.73 2) 2009 1.11 139 + 12 5 5 . 7 6 B ± 3 .28 303 + 23 7 2 . 2 8 A ± 8 .70 497 + 56 6 0 . 2 5 A ± 12.59 3) 13 .0 0 . 4 5 101 + 8 5 2 . 8 8 B ± 2 . 3 6 148 + 34 7 5 . 7 6 A ± 11 .52 263 + 69 61 . 2 1 A ± 11.82 4) 13 .0 0 .69 110 + 3 5 2 . 1 6 6 ± 0 .33 235 + 24 7 7 . 6 7 A ± 13.75 355 + 46 5 8 . 9 0 A ± 9.91 P L A S M A C O N C E N T R A T I O N R E P R E S E N T S T H E M E A N S OF S I X D E T E R M I N A T I O N S . V A L U E S W I T H D I F F E R E N T S U P E R S C R I P T 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 ) . 46 F I G U R t 8 E X P E R I M E N T 4 THE P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N IN C H I C K S F E D D I F F E R E N T D I E T S A R E SHOWN IN T A B L E S 10 AND 11 . T H E A N A L Y S I S O F V A R I A N C E OF D A T A IS P R E S E N T E D IN A P P E N D I X T A B L E 5 . T H I O U R A C I L IN C O M B I N A T I O N W I T H H Y P E R V I T A M I N O S I S E S I G N I F I C A N T L Y I N C R E A S E D T H E P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N . H Y P E R V I T A M I N O S I S E T R E A T M E N T L O W E R E D T H E P L A S M A C 0 R T I C O S T E R O N E C O N C E N T R A T I O N . T H I O U R A C I L T R E A T M E N T A L O N E HAD NO S I G N I F I C A N T E F F E C T ON P L A S M A C 0 R T I C O S T E R O N E L E V E L . TABLE 1 0 . - - P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN CH I C K S FE D D I F F E R N E N T D I E T S MA I N T A I N E D A T 1 3 C i N E X P E R I M E N T 4. C O R T I C O S T E R O N E CO N C E N T R A T I ON ( U G M S/100 M L ) 1 ' 2 AG E IN DAYS D I E T 1 D I E T 2 D I E T 3 D I E T 4 4 8 55.41 ± 4.41 ( 3 ) 52.81 ± 4.41 ( 3 ) 4 9 . 3 5 ± 8.48 ( 3 ) 6 0 . 6 0 ± 1 .22 ( 3 ) 54 3 5 . 0 8 ± 6.51 ( 2 ) 3 3 . 7 6 ± 2 . 5 9 ( 2 ) 2 7 . 2 7 ± 1.30 ( 2 ) 3 7 . 2 3 ± 6.48 ( 3 ) 57 . 2 6 . 8 4 ± 3 . 2 3 ( 3 ) 3 2 . 9 0 ± 5.33 ( 3 ) 2 5 . 9 7 ± 2.11 ( 3 ) 3 9 . 8 3 ± 1 0 . 8 8 ( 3 ) 75 3 3 . 5 9 ± 2 . 3 3 ( 3 ) 34 . 1 1 ± 6 . 0 9 " ( 3 ) 3 2 . 9 0 ± 6.48 ( 3 ) 3 8 . 9 6 ± 0.0 ( 3 ) ME A N S 3 7 . 9 3 ± 1 1 . 8 6 ( 1 1 ) 3 8 . 8 2 ± 9.90 ( 1 1 ) 3 1 . 6 0 ± 1 1 . 0 8 ( 1 1 ) 4 4 . 1 6 ± 11 . 4 6 ( 1 2 ) V A L U E S F O L L O W E D B Y S T A N D A R D DDEVI I IAT ION ( S . D . ) . V A L U E S IN P A R E N T H E S E S I N D I C A T E N U M B E R O F D E T E R M I N A T I O N S . TABLE 1 1 . - - P L A S M A C O R T I C O S T E R O N E C O N C E N T R A T I O N S IN CH I C K S FE D D I F F E R E N T D I E T S MA I N T A I N E D A T 2 4 C IN E X P E R I M E N T 4. C O R T I C O S T E R O N E CO N C E N T R A T I O N ( U G M S/100 M L ) AG E IN DAYS D I E T 1 D I E T 2 D I E T 3 D I E T 4 4 8 5 7 . 1 4 + 5.60 ( 3 ) 4 5 . 8 9 ± 3.23 ( 3 ) 45 . 0 2 ± 6.12 ( 3 ) 47 . 6 2 ± 2.44 ( 3 ) 54 3 0 . 3 0 + 8.02 ( 3 ) 4 5 . 8 9 ± 17.01 ( 3 ) 2 6 . 8 4 ± 1 0 . 8 8 ( 3 ) 5 6 . 2 8 ± 1 7 . 1 3 ( 3 ) 5 7 2 9 . 0 9 + 7.85 ( 3 ) 3 7 . 2 3 ± 3 .23 ( 3 ) 3 2 . 9 0 ± 10.67 ( 3 ) 41 .56 ± 1 4 . 6 9 ( 3 ) 7 5 2 8 . 2 2 + 0.49 ( 3 ) 2 8 . 7 4 ± 5.78 ( 3 ) 3 0 . 4 3 ± 2.11 ( 3 ) 3 0 . 3 0 ± 1.29 ( 3 ) ME A N S 3 6 . 1 9 + 1 3 . 6 5 ( 1 2 ) 3 9 . 4 3 ± 11 .68 ( 1 2 ) 3 4 . 6 3 ± 1 0 . 5 6 ( 1 2 ) 4 3 . 9 4 ± 1 4 . 7 8 ( 1 2 ) V A L U E S F O L L O W E D BY S T A N D A R D D E V I A T I O N ( S . D . ) . V A L U E S IN P A R E N T H E S E S I N D I C A T E N U M B E R O F D E T E R M I N A T I O N S . E X P E R I M E N T 5 F E E D E X P L O R A T I O N T I M E S BY L Y S I N E D E F I C I E N T AND L Y S I N E S U P P L E M E N T E D G R O U P S A R E SHOWN IN F I G U R E 9 . CH I C K S OF T H E L Y S I N E D E F I C I E N T G R O U P R E Q U I R E D MORE T I M E TO F I N D T H E F E E D T H A N D I D T H E C H I C K S F E D T H E D I E T S U P P L E M E N T E D W I T H L Y S I N E . WHEN T H E F E E D WAS M O V E D TO A NEW L O C A T I O N F O R T H E S A M E S E T OF B I R D S , I T T O O K L E S S T I M E F O R B I R D S OF B O T H G R O U P S TO L O C A T E T H E F E E D T H A N IT HAD IN T H E F I R S T T R I A L . T H E L Y S I N E D E F I C I E N T G R O U P WERE A G A I N S L O W E R IN F I N D I N G T H E F E E D T H A N WERE T H E C O N T R O L B I R D S . THE R E S U L T S OF M O V E M E N T S BY C H I C K S ON A G R I D A R E SHOWN IN A G R A P H IN F I G U R E 10. TH E C H I C K S O F L Y S I N E D E F I C I E N T G R O U P SHOWED G R E A T E R M O V E M E N T S IN E A C H T R I A L T H A N C O M P A R E D TO L Y S I N E - S U P P L E M E N T E D G R O U P O F C H I C K S . T H E P E A K O F T H E M O V E M E N T S WAS R E A C H E D ON T H E 3RD D A Y A F T E R W H I C H T I M E IT S T A R T E D D E C L I N I N G E A C H D A Y IN T H E L Y S I N E D E F I C I E N T G R O U P . |N T H E C A S E OF B I R D S F E D T H E L Y S I N E S U P P L E M E N T E D D I E T T H E P E A K OF M O V E M E N T S O C C U R R E D ON T H E 2ND D A Y AND T H E N D E C L I N E D ON S U C C E E D I N G D A Y S O F T H E T E S T P E R I O D . UN T H E 6 T H D A Y O F T H E T E S T P E R I O D T H E M O V E M E N T S A L M O S T E Q U A L L E D T H E S T A R T I N G D A Y M O V E M E N T S IN T H E L Y S I N E S U P P L E M E N T E D G R O U P O F C H I C K S . |N T H E C H I C K S O F T H E L Y S I N E D E F I C I E N T G R O U P T H E M O V E M E N T S R E M A I N E D H I G H AND WERE 3 .66 T I M E S MORE F R E Q U E N T ON T H E 6TH D A Y T H A N ON T H E S T A R T I N G D A Y . THE GROWTH R A T E S OF C H I C K S F E D L Y S I N E D E F I C I E N T AND L Y S I N E S U P P L E M E N T E D D I E T S IN E X P E R I M E N T 5 A R E SHOWN IN F I G U R E 11 . A V E R A G E W E I G H T S QF C H I C K S ON T H E L Y S I N E D E F I C I E N T D I E T WAS O N L Y 155.9 G M S . A T 39 D A Y S OF A G E , C H I C K S SHOWED C O N S I D E R A B L E W E A K N E S S AND WERE DA-LS T . M E S T A K E N BY B i R D S T O D I S C O V E R T H E L O C A T T O N O f r E E D I K E X P E R I M E N T : > . F I G U R E 10 So-\ or < Z> a oo Nr SUp CD «20-| 70J 460 ^ » DATES S//o R E S U L T S O :-IO C H I C K S OK (-.RID I N E X P E R I M E N T FIGURE 11 '.JUPPLf MI: . , i v 11 : . I. E S U P P L I E D WITH S U P P L E M E N T A L L Y S I N E AT 45 DAYS OF A G E . T H I S T R E A T M E N T R E S U L T E D IN A R A P I D GROWTH R E S P O N S E AS SHOWN I N F I G U R E 11 . DISCUSSION PLASMA CORTICOSTERONE LEVEL A NUMBER OF PHYSIOLOGICAL RESPONSES TO A DIETARY D E F I C I E N C Y OF L Y S I N E HAVE ALREADY BEEN REPORTED. HEAT PRODUCTION IS INCREASED AS A RESULT OF THE HIGHER S P E C I F I C DYNAMIC ACTION OF THE D E F I C I E N T DIET ACCORDING TO K L A I N AND WINDERS (1964) AND S W I FT _ET _A_L. (1934) AND BY INCREASED CATABOLISM OF MUSCLE PROTEIN ACCORDING TO LYMAN AND WLLCOX (1963) AND SLDRANSKY AND FARBER (1958) . PAS TRO _ET _AL . (1969) STUDIED BODY TEMPERATURE OF CHICKS FED L Y S I N E D E F I C I E N T DIETS FOR 18 HOURS, 30 HOURS, OR 2 WEEKS BEFORE MEASURING TEMPERATURES AND FOUND CONSISTENTLY HIGHER TEMPERATURES THAN IN CONTROL BIRDS FED A S I M I L A R DIET ADEQUATE IN L Y S I N E . THE PLASMA CORTICOSTERONE CONCENTRATIONS DETERMINED IN THIS STUDY WERE CONSISTENTLY HIGHER THAN HAVE BEEN REPORTED FOR AVIAN BLOOD. THE ANALYTICAL PROCEDURE DOES NOT APPEAR TO BE SATISFACTORY FOR AVIAN BLOOD. THERE IS CONSIDERABLE CONTROVERSY IN THE LITERATURE REGARDING A SATISFACTORY TECHNIQUE BUT THERE IS NO AGREEMENT ON A METHOD WHICH WILL GIVE V A L I D RESULTS. THE PLASMA C0RTICOSTERONE LEVEL IN BIRDS WAS ALWAYS HIGHER WHEN A HIGH PROTEIN (20.9$) WAS ASSOCIATED WITH A LOW L Y S I N E (0.51$) IN THE DIET THAN WHEN 2 0 . 9 $ PROTEIN WAS ASSOCIATED WITH THE REQUIRED LEVEL OF L Y S I N E ( 1 . 1 1 $ ) . CORTICOSTERONE CONCENTRATION IN PLASMA OF CHICKS FED A D I E T CONTAINING 13$ PROTEIN WERE S I M I L A R WHETHER DIETARY L Y S I N E 55 CONCENTRATION WAS 0 .65$ OR 0 .80$ AND WERE NOT S I G N I F I C A N T L Y D I F F E R E N T THAN PLASMA C0R T I C O S T E R O N E CONCENTRATION IN BIRDS FED 2 0 . 9 $ OF P R O T E I N AND 1.11$ OF L Y S I N E ( T A B L E S 6 AND 9 ) . THERE IS E V I D E N C E THAT A D M I N I S T R A T I O N OF C E R T A I N AMINO ACIDS CAN INFLUENCE THE FUNCTIONAL A C T I V I T Y OF THE ADRENAL C O R T E X . |T HAS BEEN SHOWN BY MUNRO AND MUKERJI (1958) IN RATS THAT UPTAKE OF PHOSPHORUS-32 BY L I V E R R I B O N U C L E I C ACID IS MARKEDLY S T I M U L A T E D BY G I V I N G S I N G L E LARGE DOSES OF G L Y C I N E , METHIONINE OR L E U C I N E . S I N C E THE ACT ION OF L E U C I N E AND OF METHIONINE IN TH IS R E S P E C T IS A B O L I S H E D BY ADRENALECTOMY (MUNRO AND M U K E R J I , 1962) IT WAS CONCLUDED THAT ADDIT ION OF THESE TWO AMINO ACIDS TO THE D I E T CAUSED AN INCREASED S E C R E T I O N OF ADRENOCORTICAL HORMONES, WHICH ARE KNOWN TO S T I M U L A T E L I V E R RNA METABOLISM (GOODLAD AND MUNRO , 1959) . TH IS AMINO ACID MECHANISM WAS CONFIRMED BY SHOWING THAT THE A D M I N I S T R A T I O N OF METHIONINE OR L E U C I N E TO INTACT RATS RA ISED THE CONCENTRATION OF C O R T I C O S T E R O N E IN THE P L A S M A . MUNRO _ET A L . (1963) FURTHER CONFIRMED THAT ADM INIS TRATI HAVE TH IS E F F E C T . THE A S S O C I A T I O N OF H I G H L L E V E L S OF C O R T I C O S T E R O N E WITH A HIGH L E V E L OF D I E T A R Y P R O T E I N (20.9$) WITH D E F I C I E N T L Y S I N E (0 .51$ OF THE D I E T ) CAN BE RELATED TO AMINO ACID I M B A L A N C E . A D E F I C I E N C Y OF L Y S I N E IN THE D I E T WOULD HAVE RESULTED IN A SURPLUS S U P P L Y OF OTHER AMINO A C I D S . THE INFLUENCE OF E X C E S S METHIONINE AND L E U C I N E AS CONFIRMED BY MUNRO _E_T _AL. (1963) APPEARED TO INCREASE THE C OR T I C OS TE RO NE CONCENTRATION IN THE PLASMA DEMONSTRATING THE INFLUENCE OF AMINO ACID I M B A L A N C E . AN EXCESS AMOUNT OF PROTEIN HAS GENERALLY BEEN REPORTED TO CAUSE ADRENAL HYPERTROPHY R E L A T I V E TO BODY WEIGHT. |N SOME EXPERIMENTS AN INCREASE IN FUNCTIONAL A C T I V I T Y OF THE CORTEX HAS BEEN DEMONSTRATED (MUNRO, 1 9 6 4 ) . PROTEIN D E P R I V A T I O N INDUCES THE REVERSE E F F E C T S . EVEN A SHORT PERIOD OF PROTEIN D E F I C I E N C Y LEADS TO A REDUCTION IN ADRENAL WEIGHT OUT IN PROPORTION TO LOSS IN BODY WEIGHT (MUNRO _ET _AL . , 1 9 6 2 ) . THE RESULTS OF THE PRESENT EXPERIMENTS DID NOT SHOW THAT 2 0 . 9 % PROTEIN HAD ANY S I G N I F I C A N T EFFECT ON PLASMA CORT ICOSTERONE CONCENTRATION AS COMPARED TO 1 3 % PROTEIN. IT IS PO S S I B L E THAT PLASMA C0RTICOSTERONE CONCENTRATION IS NOT AFFECTED DIRECTLY BY PROTEIN L E V E L . PLASMA CORTICOSTERONE CONCENTRATION IS RELATED TO STRESS. FIGURE 2 SHOWS THAT A NUTRITIONAL D E F I C I E N C Y OF L Y S I N E WAS RESPONSIBLE FOR HIGHER PLASMA CORTICOSTERONE CONCENTRATION AS COMPARED TO THE L Y S I N E SUPPLEMENTED CONTROL GROUP. HIGHER PLASMA C 0 R T I C O S T E R 0 N E L E V E L S HAVE BEEN OBSERVED IN RATS AFTER STARVATION BY BOULOUARD, 1 9 6 3 . CONTINUOUS STRESS COULD MODIFY THE RESPONSE OF AN ANIMAL TO STRESS. IT HAS BEEN DEMONSTRATED THAT ANIMALS REPEATEDLY EXPOSED TO STRESS SHOW A LOWER RIS E IN BLOOD C0RT ICOSTER0NE THAN UNSTRESSED CONTROLS, WHEN THEY ARE EXPOSED TO A D I F F E R E N T TYPE OF STRESS (CS A L A Y , FRENKL, 1 9 6 2 ; CSALAY _ET _AL ., 1 9 6 5 ; VECSEI AND CSALAY, 1 9 6 5 ) . THE SAME TYPE OF RESPONSE WAS OBSERVED (FIGURE 3 ) WHEN CHICKS ON L Y S I N E D E F I C I E N T DIETS WERE EXPOSED TO E L E C T R I C SHOCK ( S T R E S S E D ) , THE PLASMA CORTICOSTERONE CONCENTRATION WAS HIGHER IN CHICKS FED THE LYS INE-SUPPLEMENTED DIET WHEN THEY WERE EXPOSED TO E L E C T R I C SHOCK THAN IN CHICKS FED THE LY S I NE-DEFIC I ENT D I E T . THE EFFECTS OF THIOURACIL AND V I T A M I N E ARE SHOWN IN TABLE 10. S T A T I S T I C A L A N A L Y S I S OF DATA IS PRESENTED IN A P P E N D I X TABLE 5 . THERE WAS A S I G N I F I C A N T INCREASE IN THE PLASMA CORTICOSTERONE CONCENTRATION IN RESPONSE TO THE FEEDING OF 0 .1$ THIOURACIL IN COMBINATION WITH 220 I . U . OF V I T A M I N E . THE EXCESS V I T A M I N E BY I T S E L F LOWERED PLASMA CORTICOSTERONE CONCENTRATION. NO RESPONSE TO 0 .1$ THIOURACIL WAS OBSERVED ON CORTICOSTERONE L E V E L . THIS INDICATES THAT THERE WAS NO RE L A T I O N S H I P BETWEEN CORTICOSTERONE CONCENTRATION AND THYROID SECRETION. HOWEVER, MORRIS (1953) REPORTED THE ADRENAL HYPERTROPHY IN WHITE LEGHORN CHICKENS IN WHICH HYPOTHYROIDISM WAS INDUCED BY ADMINISTERING THIOURACIL ( 0 . 1 $ ) . FURTHER EVIDENCE TO THIS FACT HAD BEEN REPORTED BY GARREN AND SHAFFNER (1952) IN NEW HAMPSHIRE CHICKENS G I V E N THIOURACIL IN FEED AT A LEVEL OF 0 . 2 $ . AS THERE WAS NO RISE IN CORTICOSTERONE LEVEL OBSERVED BY FEEDING 0 .1$ THIOURACIL, THEREFORE, IT COULD BE PO S S I B L E THAT THE ADRENAL HYPERTROPHY OBSERVED BY MORRIS (1953) AND GARREN AND SHAFFNER (1952) WAS NOT ASSOCIATED WITH HYPER SECRETION OF THE ADRENAL CORTEX. THERE ARE SOME STUDIES OF V I T A M I N A WITH RELATION TO ADRENAL CORTEX (WOLF ET A L . , 1958; VAN DYKE _ET_AL., 1960) WHICH INDICATE THAT V I T A M I N A D I R E C T L Y INFLUENCES THE HORMONE PRODUCTION OF THE ADRENAL CORTEX TO SUCH AN EXTENT THAT SEVERE V I T A M I N A D E F I C I E N C Y WAS CONSIDERED TO REPRESENT "CHEMICAL ADRENALECTOMY" IN THE RAT. A SEARCH OF THE LITERATURE REVEALED NO EFFECT ON THE ADRENAL CORTEX OF HYPERVITAMINOSIS E. THE CHICKS HAD THE LOWER CORTICOSTER0NE CONCENTRATION WHEN THEY WERE EXPOSED TO HYPERV I TAM I NOSIS E. THIS SHOWS THAT THE HYPERVITAMINOSIS E LOWERS ADRENAL CORTEX FUNCTIONAL ACTIVITY. THE ADRENAL CORTEX FUNCTIONAL ACTIVITY WAS INCREASED SIGNIFICANTLY WITH HYPERVITAMINOSIS E AND THIOURACIL IN COMBINATION. THE MECHANISM OF ACTION ON THE ADRENAL CORTEX IS UNKNOWN. R E S P I R A T I ON RATE OXYGEN UPTAKE WAS S I G N I F I C A N T L Y HIGHER IN CHICKS R E C E I V I N G AN IMBALANCE OF DIETARY AMINO ACIDS (TABLE 7 ) . SEVERAL BIOCHEMICAL STUDIES HAD BEEN MADE OF THE EFFECTS OF GLUCOCORTICOIDS ON OXIDATIVE METABOLISM (CLARK AND PESCH 1956; GALLAGHER, 1959; GOETSCH AND MCDONALD, 1962) ON D I F F E R E N T T I S S U E S . THE RESPONSE OBSERVED DEPENDED UPON THE TYPE OF TISSU E AND DURATION OF TREATMENT. SINGLE I N J E C T I O N S OF HYDROCORTISONE HAVE BEEN FOUND TO INCREASE OXYGEN UPTAKE BY THE L I V E R (GOETSCH AND MCDONALD, 1962; GOETSCH, 1963) . IN THE PRESENT STUDY UPTAKE OF THE OXYGEN BY THE BODY AS A WHOLE WAS S I G N I F I C A N T L Y INCREASED WHEN BIRDS WERE FED A DIET CONTAINING 20.9% OF PROTEIN BUT D E F I C I E N T IN L Y S I N E ( .51%). THE SAME DIETARY TREATMENT WAS RESPONSIBLE FOR R A I S I N G THE C0RTICOSTERONE LE V E L IN THE PLASMA (T A B L E S 6 AND 9 ) . THIS INDICATED THAT THERE WAS A DIRECT R E L A T I O N S H I P BETWEEN THE CORTICOSTEROID LEVEL AND OXYGEN UPTAKE. THE EFFECT CAN BE EXPLAINED IN THAT IMBALANCE OF L Y S I N E RAISED THE CORTICOSTERONE LEVEL AND THIS INCREASE IN C0RTICOSTERONE CONCENTRATION WAS RESPONSIBLE FOR THE INCREASED CONSUMPTION OF OXYGEN BY THE BODY AS A WHOLE. HIGH UPTAKE OF OXYGEN BY VARIOUS T I S S U E S IN THE PRESENCE OF CORTICOSTERIOD HAD ALSO BEEN CONFIRMED BY BIOCHEMICAL STUDIES (CLARK AND PESCH, 1956; GALLAGHER, 1959; GOETSCH AND MCDONALD, 1962; GOETSCH, 1963) . ADRENAL WEIGHTS A WIDE V A R I E T Y OF STIMULI A P P L I E D IN YOUNG CHICKENS HAVE PRODUCED ADRENAL ENLARGEMENT ACCOMPANIED BY AN INVOLUTION OF LYMPHATIC T I S S U E . TO DETERMINE WHETHER ADRENAL ENLARGEMENT IS ASSOCIATED WITH HIGH PLASMA CORTI COSTERONE CONCENTRATIONS, ADRENAL WEIGHTS WERE RECORDED AND ANALYZED (TABLE 8 ) . THEN NUTRITIONAL STRESS OF AMINO ACID IMBALANCE RESULTED IN INCREASED PLASMA CORTICOSTERONE CONCENTRATION (TABLE 5) AND PRODUCED ADRENAL ENLARGE-MENT ( TABLE 8) ON THE BAS I S ' O F PER 100 GMS. BODY WEIGHT. ADRENAL HYPERTROPHY HAS ALSO BEEN REPORTED IN ANIMALS AFTER S E N S I T I Z A T I O N TO STRESS BY STARVATION ( B R O L I N , 1946; S E L Y E , 1936; LEBLOND AND DUGAL, 1943) . THE LITERATURE REGARDING THE EFFECT OF A C T H ON ADRENAL MASS IS C O N F L I C T I N G . SOME INVESTIGATORS HAVE FOUND NO RESPONSES EVEN TO MASSIVE DOSES, WHILE OTHERS HAVE OBSERVED HYPERTROPHY (FRANKEL E.T AL. 1 967) . BEHAVI OUR THE RESULTS OF EXPERIMENT 5 INDICATED TWO EFFECTS OF L Y S I N E D E F I C I E N C Y ( . 5 1 % ) ON THE BEHAVIOUR OF C H I C K S . THE CHICKS FED THE L Y S I N E - D E F I C I E N T DIET E X H I B I T E D MORE MOVEMENTS WHEN PLACED ON THE GRID THAN DID THE CONTROL CHICKS. WHEN CHICKS WERE STARVED FOR 1 7 HOURS AND THEN PLACED IN A NOVEL ENVIRONMENT IN WHICH FOOD WAS LOCATED AT A DISTANCE OF 1 5 FEET FROM THE POINT AT WHICH THE BIRDS WERE INTRODUCED, THE L YS 1 N E - D E F I C I E N T CHICKS WERE SLOWER IN DISCOVERING THE FOOD THAN WERE THE CONTROL CHICKS. THIS FINDING IS IN AGREEMENT WITH THE RESULTS OF A NUMBER OF STUDIES WITH RATS. OPEN F I E L D STUDIES WITH OFFSPRING OF UNDERFED MOTHER RATS REVEALED THAT THEY WERE MORE EMOTIONAL BUT HAD LESS EXPLORATORY A C T I V I T Y (S I MPSON _ET _AL . , 1 9 7 1 ) . A DIRECT R E L A T I O N S H I P HAS BEEN REPORTED BETWEEN THE S I Z E OF THE ADRENAL GLAND AND EMOTIONAL BEHAVIOUR IN RATS (ROGERS AND RICHTER, 1 9 4 8 ) WHICH WAS NOT; HOWEVER, CONFIRMED IN A SUBSEQUENT EXPERIMENT (PARE AND CULLEN, 1 9 6 5 ; ADER, 1 9 6 9 ) . RESULTS OF EXPERIMENT 5 SHOWED THAT CHICKS FED A L Y S I N E - D E F I C I E N T DIET HAD HEAVIER ADRENAL GLANDS AND SHOWED MORE EMOTION THAN CONTROL CH I C K S , INDIC A T I N G AGREEMENT WITH THE FINDINGS OF ROGERS AND RLCHTER ( 1 9 4 8 ) . THE L Y S I N E - D E F I C I E N T CHICKS IN THE PRESENT STUDY ALSO SHOWED HIGHER PLASMA CORTICOSTERONE LEVELS THAN DID THE CONTROL C H I C K S . TOUCHSTONE _ET _AL . (1 9 6 6 ) , WORKING WITH HUMAN S U B J E C T S , FOUND A P O S I T I V E CORRELATION BETWEEN THE DEGREE OF ADRENAL CORTICAL OVERACTIVITY AND DEPRESSION. THERE OCCURRED A MARKED ELEV A T I O N OF PLASMA CORTISOL AND URINARY 1 7 - O H CORTICOSTEROID LEVELS IN DEPRESSED PATIENTS CHARACTERIZED BY GREAT STRESS, ANXIETY AND A G I T A T I O N , HOWEVER, CORTISOL LEVEL WAS MUCH HIGHER ( 4 0 0 UG/1OOG) IN BR A I N T I S S U E THAN IN BLOOD (15UG/1OOML). PETERSON AND CHAIKOFF ( 1 9 6 3 ) ALSO REPORTED HIGH ACCUMULATION OF CORTISOL IN THE BRAIN OF RATS EVEN WHEN THERE WAS A VERY SMALL INCREASE IN THE CONCENTRATION IN THE BLOOD. BARNES ( 1 9 7 0 ) DISCOVERED THAT C A L O R I E - P R O T E I N MALNOURISHED BABY RATS WERE ABLE TO REGAIN WEIGHT WHEN FED A NORMAL OIET , BUT REMAINED E X C I T A B L E WHEN PLACED UNDER STRESSFUL S I T U A T I O N S . ALTHOUGH NO COMPARISONS ARE P O S S I B L E IN THE PRESENT STUDY OF THE CARRYOVER OF EARLY NUTRITIONAL EFFECTS ON BEHAVIOUR THROUGH PERIODS OF ADEQUATE NUTRITION, IT SHOULD BE NOTED THAT THE OBSERVED INCREASE IN EMOTIONALITY OF THE L Y S I N E - D E F I C I E N T BIRDS WAS APPARENT EVEN THOUGH THEY HAD BEEN FED A LYSINE-SUPPLEMENTED DIET FOR A FOUR-WEEK PERIOD DURING THE COURSE OF THE EXPERIMENTAL PERIOD. ADJUSTMENT TO STRESS CALLS FOR CHANGES IN PHYSIOLOGICAL SYSTEMS WHICH ARE NEURAL OR ENDOCRINE CONTROL, AND INVOLVES CHANGES AT CELLULAR LEVELS IN CONCENTRATION OF HORMONES OR ENZYMES AND TURNOVER OF METTABOL I TES. ACCORDING TO RENE DUBOS (1 9 6 9 ) , "GENES DOES NOT REALLY DETERMINE THE TRAITS BY WHICH WE KNOW A PERSON. THEY ONLY GOVERN THE RESPONSES THAT THE PERSON.MAKES TO ENVIRONMENTAL S T I M U L I . I N D I V I D U A L S PROGRESSIVELY EMERGE FROM THESE RESPONSES." HE FURTHER STATED THAT BRAIN DEVELOPMENT DEPENDS UPON NOURISHMENT AND ENVIRONMENT. C O N C L U S I O N S THE EFFECTS OF THE PROTEIN AND THE L Y S I N E L E V E L S ; HYPER-V I T A M I N O S I S E; AND THIOURACIL WERE STUDIED ON THE PLASMA CORTICOSTERONE CONCENTRATION IN CHICKENS. |N ADDITION, THE EFFECTS OF THE PROTEIN AND THE L Y S I N E L E V E L S WERE OBSERVED ON ADRENAL S I Z E , OXYGEN CONSUMPTION, BEHAVIOUR, AND BODY WEIGHT GAIN. DIETARY PROTEIN LEVELS OF 20.9% AND 13.0% DID NOT RESULT IN DIFFERE N T CORTICOSTER0NE CONCENTRATIONS WHEN THE DIETS CONTAINED S U F F I C I E N T L Y S I N E . THE HIGH DIETARY LE V E L OF PROTEIN (20.9%) IN CONJUNCTION WITH I N S U F F I C I E N T L Y S I N E (.51%) ALWAYS RESULTED IN HIGH PLASMA CORTICOSTERONE CONCENTRATIONS R E L A T I V E TO CONTROL. THE SAME DIETARY TREATMENT INCREASED THE ADRENAL WEIGHTS AND OXYGEN CONSUMPTION WHEN CALCULATED ON PER 100 GRAMS BODY WEIGHT B A S I S . NEITHER HYPERVITAMINOSIS E (110-220 I. U . PER KGM) NOR TEMPERATURE ( 1 3 ° C VS 2 4 ° C ) HAD ANY EFFECT ON THE PLASMA CORTICOSTERONE CONCENTRATI ON. THIOURACIL FED AT THE 0 .1% L E V E L IN COMBINATION WITH 220 I . U . OF V I T A M I N E IN THE DI E T RESULTED IN A S I G N I F I C A N T INCREASE IN THE PLASMA CORTICOSTERONE CONCENTRATION. THIS INDICATED A R E L A T I O N S H I P BETWEEN THE FUNCTION OF THE THYROID AND THE ADRENAL GLANDS. THE CHICKS FED THE L Y S I N E D E F I C I E N T D I E T HAD A DI F F E R E N T PATTERN OF BEHAVIOUR THAN THE ONES FED L Y S I N E SUPPLEMENTED D I E T . THE BEHAVIOUR EFFECT INVOLVED WAS OBSERVED THROUGH THE FEED EXPLORATION TEST AND 65 MOVEMENTS ON THE GRID. THE BIRDS FED THE L Y S I N E - D E F I C I E N T D I E T TOOK A LONGER TIME TO FIND THE FEED AND SHOWED MORE MOVEMENTS ON THE GRID THAN THOSE FED THE LYSINE-SUPPLEMENTED D I E T . THE MAXIMUM GROWTH WAS SEEN AJV I J T WEIGHTS pesr: " FROM THE BIRDS FED ON A DIET OF 20.9$ PROTEIN ASSOCIATED WITH 1.11$ L Y S I NE . 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ACTI NOMYC I N A(N(£> PU ROM YC I N I N H I B I T I O N OF CORTISONE- I NDUCED SYNTHESIS OF HEPATIC GLUCOSE - 6 PHOSPHATASE AND FRUCTOSE -I , 6 - D iPHOSPHATASE. J . B i O L . CHEM. 2 3 9 : 5 2 1 - 5 2 6 . SWANN, SwiFT, THORN, 77 WOLF, G., S. R. WAGLE, R. A. VAN DYKE AND B . C. JOHNSON, 1958. THE FUNCTION OF V I T A M I N A IN METABOLISM. I I . V I TAMIN A AND ADRENOCORTICAL HORMONES. J . B l O L . CHEM. 2 3 0 : 9 7 9 . WHITE, A., 1947. INFLUENCE OF ENDOCRINE SECRETIONS ON THE STRUCTURE AND FUNCTION OF LYMPHOID T I S S U E . HARVEY LECTURES 4 3 : 7 0 . YEH, S., J . SOLOMON AND B . F. CHOW, 1959. INFLUENCE OF V I T A M I N Bg ON T I S S U E SEROTONIN L E V E L S IN RATS. FED. PROC. 1 8 : 3 5 7 . 79 A P P E N D I X A P P E N D I X TABLE I . - - A N A L Y S I S OF VARIANCE (MEANS SQUARE) OF DATA FOR PLASMA CORTICOSTERONE IN L Y S I N E D E F I C I E N T AND L Y S I N E SUPPLEMENTED TREATMENTS. DAYS OF AGE SOURCE D.F. 21 50 61 TREATMENT 1 338.1 382.8** 99 .8 ERROR 19 120.8 3 5 . 7 273.2 INDICATES S I G N I F I C A N T DIFFERENCE (P « = 0 . 0 1 ) . APPENDIX TABLE 11 . ---ANALYS i s OF VAR iANCE FOR E L E C T R I c SHOCK RESPONSE TO PLASMA CORTICOSTERONE IN CHICKS SOURCE D.F. M.S. TREATMENT 1 13.82 SHOCKING 1 332.. 53 AGE 22. 5044.96** ERROR . 49 136.45 **|NDICATES S I GNI F I C A NOTED I FFERENCE ( P * = 0 . 0 1 ) . A P P E N D I X TABLE I I I . - - A N A L Y S I S OF VARIANCE FOR PLASMA CORTICOSTERONE CONCENTRATION IN CHICKS AT 4 5 AND 54 DAYS OF AGE IN EXPER iMENT 2. 4 5 DAYS OF AGE 54 DAYS OF AGE SOURCE D.F. M.S. D.F. M.S. TREATMENT 3 2 0 1 7 . 3 * * 1 9 9 8 . 8 ERROR 44 3 9 7 . 3 2 5 5 7 2 . 7 **|NDICATES S I G N I F I C A N T DIFFERENCE ( P < 0 . 0 1 ) . A P P E N D I X TABLE I V . - - A N A L Y S I S OF VARIANCE (MEANS SQUARE) OF PLASMA CORTICOSTERONE CONCENTRATION OF CHICKS IN EXPERIMENT 3. WEEKS OF AGE SOURCE D.F. 2 4 6 TREATMENT 3 1 2 4 . 9 * * 4 5 0 . 3 1 2 9 . 7 ERROR 2 0 2 3 . 3 4 2 8 . 0 175.6 **|NDICATES S I G N I F I C A N T DIFFERENCE ( P < = 0 . 0 1 ) . APPENDIX TABLE V . - - A N A L Y S I S OF VARIANCE OF DATA FROM EXPERIMENT 4 FOR D I E T AND TEMPERATURE INFLUENCE ON PLASMA CORTICOSTERONE CONCENTRATION. SOURCE D . F . M.S. AGE (A) TEMPERATURE (B ) THIOURACIL ( c ) V I TAMIN E (D) C X D B X C B X D A X C A X D ERROR 3 3 77 1 9 0 7 . 6 7 * * 4 . 1 5 7 9 4 . 8 9 3 9 . 9 5 * * 3 1 3 . 8 2 3.45 0.40 2 7 4 . 4 2 3 9 . 4 2 8 7 . 6 0 **|NDICATES S I G N I F I C A N T DIFFERENCE (P «= 0.01 ) . A P P E N D I X TABLE VI.--AVERAGE BODY WEIGHTS OF CHICKS FED L Y S I N E D E F I C I E N T AND L Y S I N E SUPPLEMENTED DIETS AT 40 DAYS OF AGE IN EXPERIMENT 5 . DIETARY TREATMENTS BODY WEIGHTS PROTEIN {%) L Y S I N E {%) GM. ± S. D. 1 ) 13 .0 0 .65 175.9 + 25 (10) 2) 13 .0 0 .80 400.6 + 35 (10) 1 FIGURES IN PARENTHESIS SHOW THE NUMBER OF DETERM l NAT IONS. 

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