UBC Theses and Dissertations

UBC Theses Logo

UBC Theses and Dissertations

Study of hypervitaminosis E in the chick Wong, Fook Chuen 1970-05-25

You don't seem to have a PDF reader installed, try download the pdf

Item Metadata

Download

Media
831-UBC_1970_A6_7 W65.pdf [ 5.43MB ]
Metadata
JSON: 831-1.0102205.json
JSON-LD: 831-1.0102205-ld.json
RDF/XML (Pretty): 831-1.0102205-rdf.xml
RDF/JSON: 831-1.0102205-rdf.json
Turtle: 831-1.0102205-turtle.txt
N-Triples: 831-1.0102205-rdf-ntriples.txt
Original Record: 831-1.0102205-source.json
Full Text
831-1.0102205-fulltext.txt
Citation
831-1.0102205.ris

Full Text

A STUDY OF HYPERVITAMINOSIS E IN THE CHICK BY FOOK CHUEN WONG B.S.A., NATIONAL TAIWAN UNIVERSITY, 1961 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN THE DEPARTMENT OF POULTRY SC I ENCE WE ACCENT THIS THESIS AS CONFORMING TO THE REQUIRED STANDARD THE UNIVERSITY OF BRITISH COLUMBIA OCTOBER, 1970 In presenting this thesis in partial fulfilment of the requirements for an advanced degree at the University of British Columbia, I agree that the Library shall make it freely available for reference and study. I further agree that permission for extensive copying of this thesis for scholarly purposes may be granted by the Head of my Department or by his representatives. It is understood that copying or publication of this thesis for financial gain shall not be allowed without my written permission. Department of The University of British Columbia Vancouver 8, Canada Date j£l^_^£_jllf. ACKNOWLEDGEMENT I WISH TO THANK DR. W. D. KlTTS, HEAD OF THE DEPARTMENT OF POULTRY SCIENCE, FOR PROVIDING THE FACILITY USED IN THIS STUDY. I AM GRATEFUL TO MRS. B. E. MARCH, OF THE DEPARTMENT OF POULTRY SCIENCE, WHO SUPERVISED THE PLANNING AND EXECUTION OF THE EXPERIMENTS. I ALSO THANK DRS. J. BLELY, R. FLTZSIMMONS AND M. TAIT FOR THEIR ENCOURAGEMENT THROUGHOUT MY PROGRAM. ABSTRACT A STUDY WAS MADE OF THE EFFECTS OF FEEDING EXCESS VITAMIN E TO CHICKS. THREE GROUPS OF EXPERIMENTS WERE CONDUCTED TO INVESTIGATE (1) THYROIDAL RESPONSE TO EXCESS VITAMIN E, (2) RESPIRATION RATE OF MUSCLE MITOCHONDRIA FROM CHICKS FED EXCESS VITAMIN E; AND (3) SYMPTOMS OF HYPERVITAMINOSIS E. |N TWO SEPARATE EXPERIMENTS, CHICKS WERE FED NORMAL AND EXCESS LEVELS (220 I.U./KG. OF DIET) OF VITAMIN E AND WERE SUBJECTED TO TEMPERATURES OF 14.5 AND 31.5°C. THE GOITROGENIC EFFECT OF THIOURACIL ON THE BIRDS IN THE DIFFERENT TREATMENTS WAS MEASURED. AT BOTH TEMPERATURES THE THIOURACIL-TREATED CHICKS FED EXCESS VITAMIN E EXHIBITED A LESSER ENLARGEMENT OF THE THYROID GLANDS THAN DID CHICKS RECEIVING A NORMAL LEVEL OF VITAMIN E. THIS FINDING INDICATES A REDUCTION IN THE SECRETION OF THYROID STIMULATING HORMONE (TSH) IN BIRDS FED EXCESS VITAMIN E. THE LEVEL OF VITAMIN E WHICH WAS FED DID NOT AFFECT THE GROWTH RATE OR FEED CONSUMPTION AT EITHER TEMPERATURE. I T, THEREFORE, APPEARS THAT THE METABOLIC RATES OF THE CHICKS FED THE LOW AND EXCESS LEVELS OF VITAMIN E WERE SIMILAR DESPITE DIFFERENCES IN THYROID ACTIVITY AND THAT TISSUE RESPIRATION IN BIRDS FED EXCESS VITAMIN E CAN BE MAINTAINED WITH A REDUCED SUPPLY OR TURNOVER OF THYROID HORMONE. THE ACTIVITY OF THE THYROID GLAND ITSELF WAS STUDIED IN RESPONSE TO 131 EXCESS VITAMIN E IN ANOTHER EXPERIMENT. USING RADIOIODINE (l ) AS A TRACER, IT WAS FOUND THAT THE RATES OF IODINE UPTAKE AND RELEASE BY THE THYROID GLAND WERE BOTH SLOWER IN BIRDS FED EXCESS VITAMIN E (220 I.U./KG. 0F DIET) THAN IN CONTROL BIRDS. BECAUSE VITAMIN E ACTS AS A BIOLOGICAL ANTIOXIDANT, IT MAY, IN EXCESS AMOUNTS, DEPRESS THE RATE OF OEIODINATION OF THYROXINE IN THE PERIPHERAL TISSUE AT WHICH THYROID HORMONE IS REMOVED FROM THE. CIRCULAT I ON. AS A CONSEQUENCE, THE SECRETION OF TSH WOULD BE REDUCED. IN ORDER TO OBTAIN SOME INDICATION OF CAUSE AND EFFECT IN THE MECHANISM BY WHICH THE EXCESS VITAMIN E AFFECTS THYROTROPIC HORMONE SECRETION RATE AND THYROID ACTIVITY, THE RESPIRATION RATE OF MITOCHONDRIA ISOLATED FROM THE PECTORAL MUSCLE OF CHICKS WHICH HAD BEEN FED EXCESS VITAMIN E (2200 I.U./KG. 0F DIET) WAS COMPARED WITH THAT OF THE CONTROL BIRDS. THE RESULTS SHOWED A SIGNIFICANT REDUCTION IN OXYGEN UPTAKE BY THE MUSCLE MITOCHONDRIA OF CHICKS FED THE EXCESS AMOUNT OF VITAMIN E. THE GROWTH RATE OF THE CHICKS FED THE EXCESS LEVEL OF VITAMIN E IN THIS EXPERI MENT WAS MARKEDLY LOWER THAN THAT OF THE CONTROL CHICKS. IT IS SUGGESTED THAT THE CHICK MAY MAINTAIN A NORMAL RESPIRATION RATE WHEN FED EXCESS VITAMIN E UP TO A CERTAIN LEVEL THROUGH A REDUCTION IN THYROID ACTIVITY. WITH A LARGE EXCESS OF VITAMIN E, HOWEVER, NO FURTHER COMPENSATION IS POSSIBLE. AND RESPIRATION RATE IS DEPRESSED BELOW NORMAL. EXCESS VITAMIN E CAUSED HYPOPROTHROMBINEMIA, INDICATIVE OF VITAMIN K~DEFICIENCY. OTHER SYMPTOMS NOTED WERE A REDUCTION IN HEMATOCRIT VALUES, RET ICULOCYTOSIS AND AN ABNORMALLY FL U I D- APPEARANCE OF THE BONE MARROW. BASED UPON OBSERVATIONS OF BONE CALCIFICATION, THE CALCIUM REQUIREMENT APPEARED TO BE INCREASED IN THE PRESENCE OF EXCESS VITAMIN E. TABLE OF CONTENTS PAGE LIST OF TABLES v LIST OF FIGURES vi INTRODUCTION 1 REVIEW OF LITERATURE 3 EXPERIMENT I 12 THYROIDAL RESPONSE TO EXCESS DIETARY VITAMIN E . 12 EXPERIMENT |AMATERIALS AND METHODS 1EXPER i MENT I B 14 RESULTS AND DISCUSSIONEXPERIMENT IC 20 MATERIALS AND METHODS 2RESULTS AND DISCUSSION 2 EXPERIMENT II 6 RESPIRATION RATE OF MUSCLE MITOCHONORIA FROM CHICKS FED EXCESS DIETARY VITAMIN E 2MATERIALS AND METHODS . 6 RESULTS AND DISCUSSION 9 EXPERIMENT III 35 SYMPTOMS OF KYPERVITAMINOSIS E IN CHICK 3MATERIALS AND METHODS 3RESULTS 8 DISCUSSION 44 i 11 PAGE SUMMARY 47 BIBLIOGRAPHY 9 IV LIST OF TABLES TABLE - PAGE 1A. BASAL DIETS IN EXPERIMENT |A 61 1B. BASAL DIETS IN EXPERIMENT IB2. AVERAGE THYROID WEIGHTS, BODY WEIGHTS AND FEED CONSUMPTION OF CHICKS IN EXPERIMENT IA 62 3. AVERAGE THYROID WEIGHTS, BODY WEIGHTS AND FEED CONSUMPTION OF CHICKS IN EXPERIMENT |B 3 4. ANALYSIS OF VARIANCE OF THYROIDAL RESPONSE TO THE EXCESS VITAMIN E AND/OR THIOURACIL AT DIFFERENT TEMPERATURES IN EXPERIMENT IA 64 5. ANALYSIS OF VARIANCE OF THYROIDAL RESPONSE TO EXCESS VITAMIN E AND/OR THIOURACIL AT DIFFERENT TEMPERATURES IN EXPERIMENT IB 65 6. AVERAGE THYROID WEIGHTS, BODY WEIGHTS AND FEED CONSUMPTION OF ETHOXYQUIN (EQ)-TREATED CHICKS AT 22°C IN EXPERIMENT IB 66 7. EFFECT OF EXCESS DIETARY VITAMIN E ON THYROIDAL IODINE (|131) UPTAKE AND RELEASE IN WHITE LEGHORN MALE CH I CKS 7 8. RESPIRATION RATES OF MUSCLE MITAOCHONDRIA FROM CHICKS FED THE CONTROL DIET, EXCESS VITAMIN E AND/OR THIOURACIL ... 68 9. BASAL DIET IN EXPERIMENT III 69 10. EFFECT OF EXCESS VITAMIN E ON HEMATOCRIT VALUE, PROTHROMBIN TIME AND RETICULOCYTE COUNT 70 11. PROTHROMBIN TIME OF CHICKS FED EXCESS VITAMIN E, 24 HOURS AFTER INJECTION OF MENADIONE . . 71 12. EFFECT OF EXCESS VITAMIN E ON BONE CALCIFICATION IN CHICKS FED DIETS ADEQUATE AND DEFICIENT IN CALCIUM 72 v LIST OF FIGURES FIGURE ' PAGE 1. THYROIDAL RESPONSE OF CHICKS RECEIVING TWO LEVELS OF VITAMIN E AND/OR ETHOXYQUIN TO THIOURACIL AT DIFFERENT AMBIENT TEMPERATURES IN EXPERIMENT |A 16 2. THYROIDAL RESPONSE OF CHICKS RECEIVING TWO LEVELS OF VITAMIN E AND/OR ETHOXYQUIN TO THIOURACIL AT DIFFERENT AMBIENT TEMPERATURES IN EXPERIMENT IB 17 3. EFFECT OF EXCESS DIETARY VITAMIN E ON THYROIDAL IODINE (|131) UPTAKE AND RELEASE IN WHITE LEGHORN MALE CHICKS 23 4. EFFECT OF EXCESS DIETARY VITAMIN E ON THYROIDAL IODINE (l ) UPTAKE AND RELEASE IN WHITE LEGHORN MALE CHICKS 4 5. ELECTRON MICROGRAPH OF BREAST MUSCLE MITOCHONDRIA FROM A CHICK FED A NORMAL LEVEL OF VITAMIN E 32 6. ELECTRON MICROGRAPH OF BREAST MUSCLE MITOCHONDRIA FROM A CHICK FED EXCESS VITAMIN E 33 7. ELECTRON MICROGRAPH OF BREAST MUSCLE MITOCHONDRIA FROM A CHICK FED A NORMAL LEVEL OF VITAMIN E SHOWING INTACT M I TOCHONDR I A 34 8. ERYTHROCYTES FROM A 40-DAY OLD CHICK FED A NORMAL LEVEL OF VI TAM i N E 40 9. RETICULOCYTES FROM A 40-DAY OLD CHICK FED EXCESS VITAMIN E10. ERYTHROBLAST OBSERVED ON THE BLOOD SMEAR FROM A • 40-DAY OLD CHICK 41 11. ABNORMAL BONE MARROW OF CHICKS FED EXCESS VITAMIN E 42 v I FIGURE PAGE 12A. THE INFLUENCE OF EXCESS VITAMIN E ON PERCENT OF TIBIA ASH AT DIFFERENT AGES 45 12B. THE INFLUENCE OF EXCESS VITAMIN E ON PERCENT OF TIBIA ASH RELATIVE TO BODY WEIGHT 45 v I I INTRODUCTI ON A DEFICIENCY OF VITAMIN E CAUSES A VARIETY OF SYMPTOMS IN DIFFERENT SPECIES OF ANIMALS AND MAY AFFECT MANY TISSUES (SEE REVIEW BY MASON, 1949; MATTILL, 1952; DAM, 1957; ROELS, 1967; SCOTT _ET _AL. , 1969). OTHER DIETARY COMPONENTS SUCH AS METHIONINE (CALVERT _ET _AL., 1962), SELENIUM (SCHWARZ, 1960; DESAI AND SCOTT, 1965) AND UNSATURATED FATTY ACIDS (CENTURY AND HORWITT, 1958; WITTING AND HORWITT, 1962) MAY AFFECT THE REQUIREMENT OF THE ANIMAL FOR VITAMIN E AND THE MANNER IN WHICH A DEFICIENCY OF THE VITAMIN MANIFESTS ITSELF. \T IS APPARENT THAT VITAMIN E HAS SEVERAL DIFFERENT METABOLIC FUNCTIONS BUT.THE PRIMARY MECHANISM OF ACTION OF VITAMIN E IN THE LIVING CELL HAS NOT YET BEEN DETERMINED. ONEFUNCTION OF VITAMIN E IS AS A BIOLOGICAL ANTIOXIDANT (TAPPEL, 1962). AS A RESULT OF ITS ANTIOXIDANT PROPERTIES VITAMIN E ACTS AS A NONSPECIFIC PROTECTOR OF MEMBRANE LIPIDS IN CELLS (H 0 R W I T T _ET _AL . , 1968) OR ORGANELLES SUCH AS MITOCHONDRIA (TAPPEL AND ZALK1N, 1959A), LYSOSOMES (DESAI _E_T _AL., 1964), AND MICROSOMES (TAPPEL AND ZALKIN, 1960). LACK OF SUFFICIENT TOCOPHEROL TO PREVENT UNCONTROLLED OXIDATION CAUSES THE EVENTUAL RUPTURE OF MEMBRANES AND THE RELEASE OF ENZYMES PECULIAR TO A SUBCELLULAR PARTICLE. VITAMIN E ALSO PROTECTS VITAMIN A AGAINST OXIDATION (DAM_ET_AL., 1952; PUDELK I EW I CZ _ET _AL .1964) AND HAS BEEN SHOWN TO REACTIVATE ASCORBIC ACID SYNTHESIS BY LIVER PREPARATIONS IN VITRO FROM VITAMIN E DEFICIENT ANIMALS (CAPUTTO _E_T _A_L • , 1958). THE FACT THAT THE 1 2 TOCOPHEROLS READILY UNDERGO REVERSIBLE OXIDAT I ON-REDUCTI ON, HAS LED TO THE SUGGESTION THAT OT-TOCOPHEROL MAY FUNCTION IN TERMINAL ELECTRON-TRANSFER ENZYMES AS A COENZYME OR AS A STRUCTURAL AGENT ( V AS I NG TON _ET _AL . , 1 960J DETW I LER _ET _AL . , 1966). BECAUSE VITAMIN E HAS ANTIOXIDANT PROPERTIES AND UNDERGOES REVERSIBLE OX IDAT1 ON-REDUCTI ON, IT IS POSSIBLE THAT AN EXCESS OF VITAMIN E MAY INTERFERE IN THE ACTION OF OTHER BIOLOGICAL REDOX SUBSTANCES. ALTHOUGH THERE IS A LARGE VOLUME OF LITERATURE DEALING WITH VITAMIN E DEFICIENCY, THERE HAVE BEEN FEW STUDIES REGARDING THE EFFECTS OF EXCESS VITAMIN E. SYMPTOMS OF HYPERVITAMIN0SIS E HAVE NOT BEEN DESCRIBED, NOR IS THERE EVIDENCE THAT TOCOPHEROLS PER SE EXERT ANY DELETERIOUS EFFECT IN ANIMALS OR MAN (DEMOLE, 1939; HLLLMAN, 1957). THE PRESENT INVESTIGATION WAS CONDUCTED TO DETERMINE WHAT EFFECTS EXCESS DIETARY VITAMIN E MAY HAVE IN CHIC KS. THREE GROUPS OF EXPERIMENTS WERE CARRIED OUT TO STUDY (1) THYROIDAL RESPONSE TO EXCESS DIETARY VITAMIN EJ (2) RESPIRATION RATE OF MUSCLE MITOCHONDRIA FROM CHICKS FED EXCESS VITAMIN EJ (3) SYMPTOMS OF H YPERV I TAM I NOS I S E IN "CHICKS. REVIEW OF LITERATURE GENERAL VITAMIN E WAS DISCOVERED IN 1922 BY EVANS AND BISHOP AS A FACTOR PRESENT IN VEGETABLE OIL WHICH WAS NECESSARY FOR NORMAL REPRODUCTION IN THE RAT. THE DESIGNATION OF THE LETTER E, NEXT IN ORDER FOLLOWING VITAMIN D, WAS SUGGESTED BY SURE. PURE VITAMIN E WAS FIRST ISOLATED BY EVANS AND THE EMERSONS (1936) FROM THE UNSAPONIFICABLE FRACTION OF WHEAT GERM OIL. THE NAME TOCOPHEROL WAS PROPOSED FROM THE GREEK WORD, TOKOS (OFFSPRING), PEREIN (TO BEAR), AND OL (TO SIGNIFY AN ALCOHOL). THE PREFIXES CC, (i, Y, ETC. WERE USED TO INDICATE VARIOUS FORMS OF TOCOPHEROL. AL P H A- T OC OPH E ROL", HAVING HIGH BIOLOGICAL ACTIVITY (CENTURY AND HORWITT, 1965) WAS IDENTIFIED BY FERNHOLZ (1937, 1938). DL-CC-TOCOPHERYL ACETATE IS USED AS A REFERENCE STANDARD (HUME, 1941) AND FEED SUPPLEMENT. THIS FORM OF THE VITAMIN IS NOT READILY AUTOXIDIZABLE AND HAS A HIGH BIOPOTENCY. VITAMIN E DEFICIENCY IN ANIMALS AND CHICKS HAS BEEN REPORTED BY A NUMBER OF INVESTIGATORS. V ITAMIN E DEFICIENCY CAUSES A WIDE VARIETY OF SYMPTOMS IN DIFFERENT SPECIES OF ANIMALS AND MAY AFFECT MANY DIFFERENT TISSUES, INCLUDING THE EMBRYO (FETAL RESORPTION), GONADS (STERILITY), MUSCLE ('MUSCULAR DYSTROPHY), BRAIN ( E NCE PHAL OMAL AC I A ) , BL 000 ( HEMOL YS I S OF ERYTHROCYTES, AND ANEMIA), CAPILLARY WALLS (EXUDATIVE DIATHESIS), LIVER (LIVER NECROSIS), KIDNEY (DEGENERATION OF TUBULAR EPITHELIUM), DEPOT FAT 3 4 (STEATITIS) (SEE REVIEWS BY MASON, 1949; MATTIL, 1952; DAM, 1957; ROELS, 1967; SCOTT _ET _AL. , 1969). ONE OF THE MOST WIDELY OCCURING SYMPTOMS OF VITAMIN E DEFICIENCY IN MANY ANIMALS IS MUSCULAR DYSTROPHY. AS VITAMIN E DEFICIENCY AFFECTS SUCH A WIDE VARIETY OF ANIMAL TISSUES, AND SEVERAL FACTORS SUCH AS THE DIETARY LEVELS OF METHIONINE (CALVERT ET AL., 1962) SELENIUM (SCHWARZ, 1960), AND UNSATURATED FATTY ACIDS (CENTURY AND HORWITT, 1958; WITTING AND HORWITT, 1962) INFLUENCE VITAMIN E DEFICIENCY SYNDROMES IN A VARIETY OF ANIMAL SPECIES, IT IS APPARENT THAT VITAMIN E HAS SEVERAL DIFFERENT METABOLIC FUNCTIONS, BUT NO EXACT METABOLIC ROLE OF VITAMIN E HAS YET BEEN DEFINED. VITAMIN E IS FOUND PRINCIPALLY IN THE ORGANELLA OF THE CELLS (W I SS _E_T _AL ., 1962) SUCH AS MITOCHONDRIA, MICROSOMES, AND LYSOSOMES WHERE IT IS PRESENT MAINLY IN THE REDUCED STATE (MOORE, 1959). THERE SEEMS TO BE LITTLE DOUBT THEREFORE THAT IT FUNCTIONS AS A BIOLOGICAL ANTIOXIDANT WHICH PROTECTS THE STRUCTURAL AND METABOLIC INTEGRITY OF INTRACELLULAR UNITS (TAPPEL, 1962; GUHA AND ROELS, 1965; ROELS _ET _AL., 1965; LUCY AND DINGLE, 1964). PREVENTION OF PEROXIDATION APPEARS TO BE VERY IMPORTANT IN THE MAINTENANCE OF THE STRUCTURAL INTEGRITY OF THE CELLULAR MEMBRANES OR SUBCELLULAR MEMBRANES (ZALKIN AND TAPPEL, 1960) IN TISSUES. ONE INTERESTING FACT IS THAT TOCOPHEROLS ADMINISTERED EITHER IN VIVO OR IN VITRO INHIBIT THE DIALURIC ACID-INDUCED HEMOLYSIS OF ERYTHROCYTES IN RATS (ROSE AND GYORGY, 1950, 1952). TSEN AND COLLIER (1960) AND BUNYAN _ET _AL. (I960) HAVE PRESENTED EVIDENCE THAT DIALURIC ACID PROMOTES ERYTHROCYTE HEMOLYSIS BY CATALYZING LIPID PEROXIDATION AND THAT VITAMIN E ACTS AS AN ANTIOXIDANT. C HR I S TE NSE N _E_T _AL . (1956) SHOWED THAT THE DIALURIC 5 ACID-INDUCED HEMOLYSIS MAY BE USED FOR ASSESSING VITAMIN E DEFICIENCY IN THE RAT BUT THAT IT IS NOT A RELIABLE CRITERION FOR DETERMINING A DEFICIENCY OF THIS VITAMIN IN THE CHICK. VITAMIN E MAY PLAY A ROLE IN ENERGY METABOLISM. FOR EXAMPLE, IT MAY BE A COMPONENT OF THE CYTOCHROME C REDUCTASE SYSTEMS OF THE TERMINAL RESPIRATORY CHAIN FUNCTIONING DIRECTLY AS AN ELECTRON CARRIER (BIOLOGICAL OXIDATION AND REDUCTION) OR INDIRECTLY AS A BINDING AGENT (VASINGTON, REICHARD AND NASON, 1960). OL I VER I RA _ET _AL . ( 1 96 9) DEMONSTRATED THAT CC- TOCOPHEROL DID NOT HAVE A DIRECT QUANTITATIVE RELATIONSHIP WITH ENZYMATIC ACTIVITIES. |T HAS ALSO BEEN SUGGESTED THAT IT MAY FUNCTION DIRECTLY OR INDIRECTLY IN NORMAL OXIDATIVE PHOSPHORYLATION IN TISSUE RESPIRATION (VASINGTON, REICHARD AND NASON, 1960), OR IN THE METABOLISM OF COENZYMEQ(GREEN, 1962). THYROIDAL RESPONSE TO TEMPERATURE AND DIET ALTHOUGH THE MECHANISM OF ACTION OF THYROID HORMONES STILL REMAINS UNKNOWN, THE THYROID GLAND IS CONSIDERED TO PLAY AN IMPORTANT ROLE IN THE ADAPTIVE PHENOMENA ASSOCIATED WITH CHANGES IN EXTERNAL ENVIRONMENT AND HAS BEEN THE SUBJECT OF NUMEROUS INVESTIGATIONS IN THIS REGARD. |N THESE STUDIES, THYROID ACTIVITY HAS BEEN EVALUATED BY DIFFERENT METHODS UTILIZING A VARIETY OF PARAMETERS SUCH AS BASAL METABOLIC RATE, THYROID WEIGHT, HISTOLOGICAL APPEARANCE OF THE TISSUE, PROTEIN-BOUND IODINE LEVELS, RADIOIODINE UPTAKE AND THYROIDAL IODINE TURNOVER RATE. MANY.FACTORS SUCH AS GENETIC STRAIN, AGE, ENVIRONMENTAL TEMPERATURE, AND DIETARY INTAKE OF IODINE ARE KNOWN TO AFFECT THYROID ACTIVITY. HOWEVER, COMPARATIVELY LITTLE 6 IS KNOWN ABOUT THE EFFECTS OF NUTRITIONAL FACTORS OTHER THAN IODINE ON THYROID SECRETION RATE. THERE IS MUCH DATA TO INDICATE THAT THE THYROID GLAND OF BIRDS RESPONDS TO ENVIRONMENTAL TEMPERATURE CHANGES (HOFFMANN AND SHAFFNER, 1950; JOINER AND HUSTON, 1957; PREMACHANDRA _ET _AL ., 1958; HEN I NGER ET AL ., 1960; STAHL AND TURNER, 1961; HUSTON _E_T _AL. , 1962). THE YOUNG BIRD HELD IN A COOL ENVIRONMENTAL TEMPERATURE WAS SHOWN TO HAVE HEAVIER THYROID GLANDS AND TO SECRETE MORE THYROXINE THAN THOSE EXPOSED TO HIGH TEMPERATURES (HOFFMANN AND SHAFFNER, 1950). LATER REPORTS (JOINER AND HUSTON, 1957; HUSTON ET M.., 1962) CONFIRMED THESE FINDINGS. HENDRICH AND TURNER (1963) ALSO DEMONSTRATED THAT SHORT TERM EXPOSURE TO COLD ,131 STIMULATED THE THYROID OF THE FOWL AS INDICATED BY THYROIDAL !. UPTAKE AND RELEASE WAS SHOWN TO RETURN TO NORMAL BY LONGER TERM EXPOSURE TO COLO. A STUDY BY HENDRICH AND TURNER (1965) DEMONSTRATED THAT A CHANGE FROM SUMMER TEMPERATURE TO CONSTANT 4.4°C COLD HAD NO SIGNIFICANT EFFECT ON THE T2 OF THYROXINE-I IN FOWLS AND SUGGESTED THAT FLUCTUATING ENVIRONMENTAL TEMPERATURES PRODUCE CHANGES IN THE THYROID HORMONE UTILIZATION. KOBAYASHI AND GORBMAN (i960) AND ROSENBERG _ET _A_L. (1964) REPORTED ,131 THAT THE TIME OF PEAK I -UPTAKE WAS INFLUENCED BY THE CONCENTRATION OF IODIDE IN THE DIET. KOBAYASHI AND GORBMAN (i960) DEMONSTRATED THAT WITH A LOW IODINE DIET, RADIOACTIVITY FELL RAPIDLY AFTER REACHING A PEAK AT 6 HOURS. VERY LITTLE IS KNOWN OF THE RELATION OF THYROID FUNCTION AND J 31 " c VITAMIN E. THE RATE OF I UPTAKE BY THE THYROID IN VITAMIN E EXCESS HAS BEEN REPORTED TO BE REDUCED IN THE RAT (VALENTI AND BOTTARELLI , 1965) AND 7 A SIMILAR OBSERVATION IN HAMSTER WAS REPORTED (DURAND _ET _AL ., 1968). |F A COMPOUND IS ADMINISTERED WHICH INHIBITS THE SYNTHESIS OF THYROID HORMONES, A COMPENSATORY HYPERTROPHY OF THE THYROID GLAND OCCURS IN RESPONSE TO AN ELEVATION IN TH YROTROPIF I C HORMONE (TSH) SECRETION RATE. IN ANIMALS SUBJECTED TO DIFFERENT ENVIRONMENTAL TEMPERATURES, THE RESULTANT DIFFERENCES IN SECRETION RATES OF TSH (HENDRICH AND TURNER, 1964) AND THYROID HORMONE WILL BE REFLECTED IN DIFFERENCES IN THE GOITROGENIC RESPONSE TO ADMINISTRATION OF A GOITROGENIC COMPOUND SUCH AS THIOURACIL. THIOURACIL EXERTS ITS EFFECT BY PREVENTING THE FORMATION OF ORGANIC IODINATED COMPOUNDS IN THE THYROID GLAND AND PREVENTING RECYCLING OF IODINE (FRANDLIN, LERNER AND CHAIKOFF, 1944; ASTWOOD, 1945; VANDERLAAN AND BLSSEL, 1946). KOBAYASHI AND GORBMAN (I960) OBSERVED IN CHICKS THAT THE RATIO OF MONO I ODOT YRO.S I NE (MIT) TO DL I OD 0 T YROS I NE (DIT) WAS GREATER AT 46 HOURS AFTER ADMINISTRATION OF THIOURACIL THAN AT 6 HOURS, INDICATING ACCUMULATION OF MIT, AND SUGGESTED THAT THIOURACIL PRODUCED A BLOCK IN THYROXINE SYNTHESIS BETWEEN MIT AND DIT. IN RECENT YEARS, SEVERAL INVESTIGATORS (CUNNINGHAM, 1964; MALOOF AND SOODAF, 1963J JLROUSEK AND CUNNINGHAM, 1968), SUGGESTED THAT A PROTEIN SULFENYL IODIDE AS AN ACTIVE CARRIER FOR |+ IS A KEY INTERMEDIATE IN THE ENZYMATIC IODINATION OF TYROSINE IN THE THYROID GLAND. THE SULFENYL IODIDE INTERMEDIATE SERVES TO STABILIZE I WHICH IS INVOLVED IN THE IODINATION OF TYROSINE. A POSSIBLE MECHANISM IS THAT ANTITHYROID AGENTS SUCH AS THIOURACIL AND THIOUREA HAVE VERY HIGH RATES OF REACTION WITH SULFENYL IODIDE TO FORM AN INACTIVE MIXED DISULFIDE (CUNNINGHAM, 1964). • THIOURACIL NOT ONLY BLOCKS THE FORMATION OF THYROXINE IN THE THYROID 8 BARKER _ET _AL. ' (1 949), AND I K _ET _AJ_ . (1949), AND STASILLI ET AL. (1960) SHOWED THAT THE CALORIGENIC EFFECTIVENESS OF THYROXINE WAS DECREASED BY FEEDING EITHER THIOURACIL OR METHYLTHIOURACIL. THE RESULTS OF SEVERAL REPORTS (VAN ARSDEL AND WILLIAMS, 1956; HOGNESS _ET _AL ., 1954; STASILLI _E_T .AL., 1960; JONES AND M I DDLESWORTH, 1960; ESCOBAR AND ESCOBAR, 1961, 1962; HERRERA _ET A_L ., 1963) HAVE SHOWN THAT PROPYLTHIOURACIL ,131 DECREASED THE AMOUNT OF I EXCRETED IN THE URINE AND INCREASED THE ,131 ,131 FECAL EXCRETION OF I -THYROXINE AFTER THE ADMINISTRATION OF I LABELED THYROXINE, PRESUMABLY BY INHIBITING THEIR DEIODINAT I ON. DEIODINATION OF BOTH PHENOLIC RING AND INNER RING IODINES IS AFFECTED (FLOCK AND BOLLMAN, 1962; HERRERA _ET .AL., 1963). JAGIELLOW AND MCKENZIE (1960) HAVE OBSERVED THAT THE PROTEIN-BOUND IODINE OF THIOURAC IL-TREATED ANIMALS GIVEN GO I TER-PREVENT1NG DOSES OF THYROXINE WAS HIGHER THAN THAT OF THYROXINE-TREATED, THYROIDECTOM I ZED ANIMALS. HERSHMAN AND VAN MLDDLESWORTH (1962) SUPPORTED THESE FINDINGS. IT HAS BEEN SUGGESTED THAT THIOURACIL AND PROPYLTHIOURACIL ENHANCE THE BINDING OF THE HORMONE BY THE PLASMA PROTEIN, THEREBY DECREASING THE PERIPHERAL ACTIVITY OF THYROXINE (HERSHMAN AND VAN MIDDLESWORTH, 1962). RADIOIODINE UPTAKE 1 31 THE DISCOVERY OF RADIOIODINE I BY ENRICO FERMI IN 1935 LED TO ITS USE BY HAMILTON AND SOLEY IN 1939 IN THE TEST OF THYROID FUNCTION. RADIOIODINE UPTAKE AS A PARAMETER OF THYROID ACTIVITY IS BASED UPON THE DYNAMICS OF IODINE METABOLISM WHICH MAY BE CONSIDERED FROM THREE ASPECTS: 9 (1) THE INORGANIC IODIDE IS TAKEN UP RAPIDLY FROM THE CIRCULATING BLOOD (VANDERLAAN AND VANDERLAAN, 1947; TAUROG _ET _AL ., 1947; HALMI, 1961); (2) IODINE MAY BE ORGANICALLY BOUND AS EARLY AS 15 MINUTES AFTER ADMINISTRATION (C HALKOFF AND TAUROG, 1949; WLLLIAMS AND VLCKERY, 1965); (3) THE ORGANICALLY BOUND RADIOIODINE IS SOON RELEASED FROM THE THYROID GLAND (NADL'E-R AND LEBLOND, 1955 AND 1958). A 24-HOUR-UPTAKE TEST IS WIDELY USED AND IS BASED ON THE ASSUMPTION THAT THE MAXIMUM LEVEL OF ,131 ,131 THYROIDAL I ACCUMULATION IS REPRESENTED BY THE.LEVEL OF THYROIDAL I 131 24 HOURS AFTER ADMINISTRATION OF I (WAHLBERG, 1955; HAMILTON AND SOLEY, 1940; ROSENBERG _ET _AL. , 1963). SOME INVESTIGATORS COMMENT THAT SINCE (1) T«M E R E IS RELEASE OF INORGANIC IODINE FROM THE GLAND (HALMI AND PLTT-RLVERS, 1962); (2) RADIOIODINE IN THE THYROID GLAND IS EASILY AFFECTED BY THE LEVEL OF IODINE IN THE DIET (ROS E N BE RG _ET _AL . , 1964); (3) IODINE IS METABOLIZED IN THE THYROID GLAND IN A HETEROGENEOUS MANNER (ROSENBERG ET AL., 1966); THE VALIDITY OF UPTAKE OR RELEASE MEASUREMENTS AS AN INDEX OF . 131 THYROID ACTIVITY ACCORDINGLY BECOMES QUESTIONABLE. NEVERTHELESS, I UPTAKE AND RELEASE BY THE THYROID ARE EMPLOYED RATHER WIDELY AS COMPARATIVE INDICES OF THYROID ACTIVITY SINCE THE SPECIFIC RADIOACTIVITY OF THE IODINE DEMONSTRATED IN THE THYROID TISSUE IS EASILY MODIFIED BY CIRCUMSTANCES WITHIN THE NORMAL VARIATION OF THE PHYSIOLOGY OF THE ANIMALS. THE UPTAKE OF RADIOIODINE BY THE GLAND DOES NOT INDICATE THE ABSOLUTE AMOUNT OF IODINE PICKED UP, BUT MERELY REPRESENTS THE PERCENTAGE OF THE TOTAL STABLE IODIDE POOL TRAPPED (S I L VE R JET _AL . , 1955; ANDREWS, 1957). 10 VITAMIN E AND OXYGEN CONSUMPTION OF TISSUES VITAMIN E DEFICIENCY HAS BEEN SHOWN TO INFLUENCE OXYGEN CONSUMPTION BUT THE EFFECT VARIES WITH THE TYPE OF TISSUE STUDIED. MARKED INCREASES IN OXYGEN CONSUMPTION FROM VARIOUS SPECIES OF VITAMIN E DEFICIENT ANIMALS HAVE BEEN REPORTED (VICTOR, 1934J MADSEN, 1936; FRIEDMAN AND MATTILL, 1941; HOUCHIN AND MATTILL, 1942A,B,C; HOUCHIN, 1942; KAUNITZ AND PAPPENHE IMER, 1943; RODERUCK _ET _AL., 1949; HIMMEL AND MELVILLE, 1951; ROSENKRANTZ, 1955; BIRD_ET_AL., 1963). RABBITS, HAMSTERS, RATS, AND GUINEA PIGS WERE USED AS THE EXPERIMENTAL ANIMALS IN THESE STUDIES. VICTOR (1934) OBSERVED NO EFFECT OF VITAMIN E DEFICIENCY ON THE OXYGEN CONSUMPTION OF DUCK MUSCLE. ROSENKRANTZ (1955) FOUND THAT RABBIT ADRENAL CORTEX AND LIVER SLICES SHOWED A DEFINITE INCREASE IN OXYGEN CONSUMPTION WHEREAS OXYGEN CONSUMPTION OF HEART. AND KIDNEY TISSUES REMAINED UNCHANGED IN VITAMIN E DEFICIENCY. GRIGORYEVA AND SHCHUKINA (1967) OBSERVED THAT OXYGEN CONSUMPTION FOR THE OXIDATION OF DR KETO-GLUTARIC AND SUCCINIC ACIDS PER MG OF PROTEIN IN MITOCHONDRIA FROM DYSTROPHIC MUSCLE IN VITAMIN E DEFICIENCY WAS SIMILAR TO THAT FROM NORMAL TISSUE. VITAMIN E AND MITOCHONDRIA MITOCHONDRIA CONTAIN MUCH OF THE VITAMIN E PRESENT IN CELLS (WlSS _ET _AL., 1962). OL I VE I RA E_T _AL . (1968) FOUND TOCOPHEROL TO BE PRESENT -IN ALL THE ENZYMATICALLY ACTIVE LIPO-PROTEIN COMPLEXES OF THE ELECTRON TRANSPORT SYSTEM OF INNER MITOCHONDRIA. FURTHER, THEY DEMONSTRATED THAT THE TOCOPHEROL DID NOT APPEAR TO HAVE A DIRECT QUANTITATIVE RELATIONSHIP WITH THE ACTIVITY OF ENZYMES, SUCH AS CYTOCHROME OXIDASE, NADH- AND , SUCCINATE-CYTOCHROME C REDUCTASES. MITOCHONDRIA CONTAIN ABOUT 25$ LIPID 11 (SWANSON AND ARTOM, 1950), MUCH OF WHICH IS UNSATURATED AND GREATER THAN 90$ is PHOSPHOLIPID. MOST OF THE LIPID OCCURS IN THE MITOCHONDRIAL MEMBRANE (FLEISCHER"AND ROUSER, 1965j FLEISCHER, 1967j CHAPMAN AND LESLIE, 1970). SALKIN AND TAPPEL (i960) REPORTED THAT IN VIVO PEROXIDATION OCCURS IN THE LIVER AND KIDNEY OF VITAMIN E DEFICIENT RABBITS. LIPID PEROXIDATION IS CATALYZED BY HEMATIN COMPOUNDS, SUCH AS THE CYTOCHROMES CONTAINED IN MITOCHONDRIA (TAPPEL AND ZALKIN, 1959B). AN ELECTRON MICROSCOPIC STUDY HAS SHOWN MITOCHONDRIAL ALTERATIONS IN SKELETAL MUSCLE OF YOUNG CHICKS FED A VITAMIN E DEFICIENT DIET (CHEVILLE, 1966). FURTHER, VAN VLEET ET AL• (1968) SUGGESTED THAT TISSUE IN WHICH VITAMIN.E HAS BEEN DEPLETED, SKELETAL MUSCLE MITOCHONDRIA UNDERWENT LIPID PEROXIDATION WITH RESULTING DESTRUCTION OF THE INNER MEMBRANES. FRAGMENTATION OF CRISTAE, MITOCHONDRIAL SWELLING DUE TO ALTERED MEMBRANE PERMEABILITY, FORMATION OF 1 NTRAM I TOCHONDRIAL MYELIN-FIGURE-L I KE MEMBRANOUS PROFILES, AND ACCUMULATION OF I NTRAM I TO-CHONDRIAL DENSE GRANULES RESULTED. EXPERIMENT 1 THYROIDAL RESPONSE TO EXCESS DIETARY VITAMIN E TWO EXPERIMENTS WERE CONDUCTED TO STUDY THE EFFECTS OF EXCESS VITAMIN E IN RELATION TO THYROID ACTIVITY. |N ONE EXPERIMENT,- CHICKS WERE FED NORMAL AND EXCESS LEVELS OF VITAMIN E AND WERE SUBJECTED TO DIFFERENT TEMPERATURES. THE GOITROGENIC EFFECT OF THIOURACIL WAS MEASURED. |N A ,131 SECOND EXPERIMENT, THE RATES OF I UPTAKE AND RELEASE WERE COMPARED IN CHICKS WITH NORMAL AND EXCESS INTAKE OF VITAMIN E. . EXPERIMENT 1A MATERIALS AND METHODS THIS EXPERIMENT WAS STARTED IN NOVEMBER, 1969. EIGHTY DAY-OLD MALE WHITE LEGHORN CHICKS WERE WING-BANDED AND DISTRIBUTED AT RANDOM INTO 3 GROUPS IN ELECTRICALLY HEATED BATTERY BROODERS. THE CONSTITUENTS OF BASAL EXPERIMENTAL DIET IS SHOWN IN TABLE 1A. GROUP A RECEIVED ONLY THE BASAL DIET (WITHOUT SUPPLEMENTARY VITAMIN E), GROUP B RECEIVED AN EXCESS VITAMIN E LEVEL OF 220 I.U. PER KG. OF DIET AND GROUP C RECEIVED THE BASAL DIET PLUS ETHOXYQUIN^ (EQ) AT 0.25$ OF THE DIET. ROOM TEMPERATURE WAS THERMOSTATICALLY CONTROLLED AT 22°C. THE EXPERIMENTAL DIETS AND WATER WERE FED AD LIBITUM. 1,2 DIH YD R0-6-ETH0XY-2, 2, 4-TRI METHYLQUINOLINE. 12 13 AT 23 DAYS OF AGE, ALL THE BIRDS OF THE 3 GROUPS WERE WEIGHED AND 24 CHICKS FROM EACH GROUP WERE SELECTED BY A SYSTEMATIC PROCEDURE ACCORDING TO THE INDIVIDUAL BODY WEIGHTS IN ORDER TO REDUCE THE VARIATIONS OF THE INITIAL BODY WEIGHTS OF THE EXPERIMENTAL CHICKS AMONG THE 3 GROUPS. EACH GROUP WAS DIVIDED INTO 2 SUBGROUPS FOR THE HOT AND COLD ENVIRONMENTAL CHAMBERS. THE COLD ENVIRONMENTAL CHAMBER WAS REGULATED AT 14.5°C AND THE HOT ENVIRONMENTAL CHAMBER AT 31.5°C. THE VARIOUS GROUPS OF. CHICKS WERE PLACED IN THE ENVIRONMENTAL CHAMBERS FOR 2 DAYS ADAPTATION BEFORE THE EXPERIMENT WAS STARTED. EACH SUBGROUP WAS DIVIDED INTO 2 LOTS OF 6 CHICKS EACH. ONE LOT WAS MAINTAINED ON THE ORIGINAL EXPERIMENTAL DIET AND THE OTHER LOT RECEIVED 0.1$ THIOURACIL ( TU ) IN THE EXPERIMENTAL DIET. THE TREATMENTS ARE SUMMARIZED AS FOLLOWS: . EXPERI MENTAL PES I GN EXCESS EXCESS VITAMIN E Q BASAL TU Vi TAM I N E +JJJ EQ +TU 14.5°C 6* 6 6 6 6 6 31.5°C 6 6 6 6 6 6 *NUMBER OF CHICKS. THE TEMPERATURES IN THE 2 ENVIRONMENTAL CHAMBERS WERE RECORDED BY A RECORDING THERMOGRAPH. FEED CONSUMPTION WAS ALSO RECORDED. THE EXPERIMENTAL PERIOD WAS 14 DAYS. AT THE END OF THE EXPERIMENT, ALL CHICKS WERE WEIGHED AND KILLED. THE THYROID GLANDS WERE REMOVED AND WEIGHED. 14 EXPERIMENT 1B THIS EXPERIMENT WAS STARTED IN JULY, 1970. THE PROCEDURE AND THE TREATMENTS WERE THE SAME AS EXPERIMENT 1A WITH EXCEPTION OF LARGER POPULATION (15 INDIVIDUALS) IN EACH LOT AND TEMPERATURE (22°C) IN THE ETHOXYQU I N-TREATED BIRDS. THE BASAL DIET WAS ALSO MODIFIED AS SHOWN IN TABLE 1B, BECAUSE THE DIETS FED IN EXPERIMENT 1A WERE AC CIDENTLY FORMULATED WITHOUT ADEQUATE CALCIUM SUPPLEMENTATION. RESULTS AND DISCUSSION . THE RESULTS FOR EXPERIMENTS 1A AND 1B ARE SUMMARIZED IN TABLES 2 AND 3. AVERAGE VALUES ARE EXPRESSED ± SAMPLE STANDARD DEVIATION. IN BOTH EXPERIMENTS, THE THYROID WEIGHTS OF THE CHICKS PER 100 GM. BODY WEIGHT WERE SIGNIFICANTLY GREATER IN THE 14.5°C ENVIRONMENTAL CHAMBER THAN THOSE IN THE 31 .5°C ENVIRONMENTAL CHAMBER (P sgO.05). THIS RESULT IS IN AGREEMENT WITH PREVIOUSLY REPORTED FINDINGS (HOFFMANN AND SHAFFNER, 1950; JOINER AND HUSTON, 1957; HUSTON _ET _AL. , 1962). THIOURACIL CAUSES GOITER BY ENHANCING TSH OUTPUT IN RESPONSE TO A BLOCK IN THE SYNTHESIS OF THYROXINE BY THE THYROID GLAND (DEMPSEY AND ASTWOOD, 1943). THE EXTENT OF WHICH THYROID WEIGHT INCREASES IN RESPONSE TO THE FEEDING OF THIOURACIL VARIES WITH THE RATE OF SECRETION OF TSH (BARKER, 1955; BROWN-GRANT, 1957). THE RATE OF SECRETION OF TSH is, IN TURN,•DEPENDENT UPON THE NORMAL RATE AT WHICH THYROXINE IS SECRETED BY THE ANIMAL. ACCORDINGLY THE RESPONSE OF THE THYROID GLAND TO THIOURACIL IN THE PRESENT EXPERIMENTS IS CONSIDERED TO BE AN INDICATION OF THE RATE OF THYROXINE SECRETION UNDER THE EXPERIMENTAL TREATMENTS IMPOSED, '15 I.E. DIFFERENT ENVIRONMENTAL TEMPERATURES AND DIETARY LEVELS OF VITAMIN E. EXCESS VITAMIN E DID NOT AFFECT THE THYROID SIZE OF THE CHICKS FED THE DIET WITHOUT THIOURACIL AT EITHER TEMPERATURE (TABLES 2 AND 3, FIGURES 1 AND 2). THYROID SIZE IS NOT A PARTICULARLY SENSITIVE PARAMETER OF THYROID ACTIVITY OR TSH SECRETION RATE. IT IS KNOWN THAT HIGHER LEVELS OF TSH ARE REQUIRED TO INCREASE THYROID SIZE THAN IS NEEDED TO INCREASE THYROXINE SECRETION RATE. EXCESS VITAMIN E DID AFFECT THYROID WEIGHT OF THE THIOURACIL-TREATED CHICKS (FIGURES 1 AND 2). IN EXPERIMENT 1A, THE THIOURACIL-TREATED CHICKS FED EXCESS VITAMIN E EXHIBITED A LESSER THYROID RESPONSE THAN DID THE THIOURACIL-TREATED CHICKS FED THE LOW VITAMIN E. THE DIFFERENCE IN RESPONSE WAS NOT STATISTICALLY SIGNIFICANT (TABLE 4) AND MAY HAVE BEEN DUB TO SMALL NUMBERS OF CHICKS EMPLOYED IN THE EXPERIMENT. THE EXPERIMENT WAS, THEREFORE, REPEATED WITH A LARGER POPULATION. THE RESULTS OF THIS SECOND EXPERIMENT CONFIRMED THE EVIDENCE OF EXPERIMENT 1A. THE DIFFERENCE IN THYROIDAL RESPONSE OF CHICKS FED THE DIFFERENT LEVELS OF VITAMIN E WAS STATISTICALLY SIGNIFICANT (P ^ 0.01) AS SHOWN IN TABLE 5. IT WAS CONCLUDED THAT THE RATE OF TSH SECRETION IN THE CHICKS FED EXCESS VITAMIN E WAS LESS THAN THAT IN THE CHICKS FED THE DIETS WITHOUT VITAMIN E SUPPLEMENTATION AND AS A CONSEQUENCE, EXCESS VITAMIN E INDIRECTLY DEPRESSED THYROID ACT I V ITY. ONE HYPOTHESIS CONSIDERED FOR THYROID RESPONSE TO DIETARY VITAMIN E WAS BASED ON THE FACT THAT LOW VITAMIN E CAUSED A DECREASE IN CIRCULATING THYROXINE LEVEL DUE TO INCREASED PERIPHERAL D EIOD INA TI ON IN THE LIVER AND MUSCLE (GALTON AND HlNGBAR, 1965). GALTON AND HlNGBAR (1965) OBSERVED BASAL EXCESS vITAMIN E 0.25$ ETHOXYQUIN 31 14.5UC 31.5UC 14.5°C 14.5°C 31 .5^ 14.5UC 31 .5°C WITHOUT THIOURACIL IN THE DIET WITH THIOURACIL IN THE DIET FIG. 1. THYROID RESPONSE OF CHICKS RECEIVING TWO LEVELS OF VITAMIN E AND/OR ETHOXYQUIN TO THIOURACIL AT DIFFERENT AMBIENT TEMPERATURES IN EXPERIMENT 1A. 50 40 30 20 10 17 BASAL EXCESS vITAMiN E 0.25$ ETHOXYQU1N 14.5°C 31,5°C 22°C 14.5°C 31.5°C 22°C WITHOUT THIOURACIL IN THE DIET WITH THIOURACIL IN THE DIET FIG. 2. THYROIDAL RESPONSE OF CHICKS RECEIVING TWO LEVELS OF VITAMIN E AND/OR ETHOXYQUIN TO THIOURACIL AT DIFFERENT AMBIENT TEMPERATURES IN EXPERIMENT 1B. 18 THAT DE10DINATI0N OF THYROXINE V/AS INCREASED IN HOMOGENATES OF MUSCLE AND LIVERS FROM VITAMIN E DEFICIENT ANIMALS AND THAT LARGE DOSES OF TOCOPHEROL, EITHER ADM INISTERED IN VIVO OR ADDED TO TISSUE HOMOGENATES IN VITRO, GREATLY DECREASED HEPATIC OR MUSCULAR DE IOD I NAT ING ACTIVITY IN BOTH NORMAL AND VITAMIN E DEFICIENT RATS. GRUENSTEIN (1970) HAS RECENTLY PROPOSED A THEORY FOR THE MECHANISM OF ACTION OF THYROXINE. HE SUGGESTS THAT THYROXINE ASSOCIATES WITH THE LIPIDS OF THE CELL MEMBRANE, WHEREUPON IT IS DEGRADED BY A FREE RADICAL MECHANISM, RESULTING IN THE RELEASE OF SOME OR ALL OF THE IODINE IN THE + 0 / FORM OF I OR I . SEVERAL INVESTIGATORS (ROSENBERG AND JENDRASIAK, 1968; F l NDELSTE IN "AND CASS, 1968) HAD PREVIOUSLY SUGGESTED THAT A POSITIVE ION, I OR A FREE RADICAL I , RELEASED FROM THE THYROXINE MAY INTERACT WITH PHOSPHOLIPIDS AND CHOLESTEROL OF THE MEMBRANE, CHANGING THE PHYSICAL, STRUCTURAL AND/OR CHEMICAL PROPERTIES OF THESE LIPIDS SO THAT THE ELECTRICAL RESISTANCE OF THE MEMBRANE IS DECREASED. THE STUDIES OF WYNN (1968A,B) ON THYROXINE DEGRADATION IN RAT LIVER HAVE SHOWN THAT SOME OR ALL OF THE IODINE OF THYROXINE IS RELEASED AS PART OF THE INITIAL METABOLISM OF THE HORMONE AFTER IT REACHES TARGET TISSUES FROM THE PLASMA. HE FURTHER DEMONSTRATED THAT THYROXINE FUNCTIONS AS AN EXTREMELY POTENT ANTIOXIDANT, CAPABLE OF PREVENTING PEROXIDATION OF LECITHIN DERIVED FROM THE MICROSOMAL MEMBRANE AT EVEN LOWER CONCENTRATIONS THAN VITAMIN E (WYNN, 1968A). CASH ET AL. (1966) REPORTED THE ANTIOXIDANT PROPERTIES OF THYROXINE IN PREVENTING THE PEROXIDATION OF PHOSPHOLIPIDS IN MITOCHONDRIAL MEMBRANE. WYNN (1968A) HAS SHOWN THAT DURING THE ANT I OX I DAT I ON REACTION IN THE PRESENCE OF METAL ION (FE ), OXYGEN AND PHOSPHOLIPID, THYROXINE IS ITSELF DEGRADED^ TO 19 RELEASE ITS IODINE. V I TAMI N E IS GENERALLY'CONSIDERED TO BE A BIOLOGICAL ANTIOXIDANT. THE RESULTS OF THE PRESENT EXPERIMENTS SUGGEST THAT EXCESS VITAMIN E MAY REPLACE THYROXINE IN ITS. ANTIOXIDANT FUNCTION AND D EIOD I ANT I ON OF THYROXINE MIGHT, THEREFORE, BE RETARDED. THE DEPRESSION OF DEIODINATION OF. THYROXINE COULD THEN REDUCE THE RATE AT WHICH THYROID HORMONE IS REMOVED FROM THE CIRCULATION, THUS MAINTAINING THE LEVEL OF THE HORMONE IN THE CIRCULATION AND, THEREFORE, SUPPRESSING SECRETION OF TSH. GROWTH RATE AND FEED CONSUMPTION WERE SIMILAR FOR LOW AND EXCESS VITAMIN E TREATED BIRDS KEPT AT TWO TEMPERATURES. THESE RESULTS INDICATE THAT METABOLIC RATES WERE SIMILAR IN BIRDS ON BOTH TREATMENTS DESPITE DIFFERENCES IN THYROID ACTIVITY. TH IS SHOULD SUGGEST THAT TISSUE RESPIRATION IN BIRDS FED EXCESS VITAMIN E CAN BE MAINTAINED WITH A REDUCED SUPPLY OR TURNOVER OF THYROXINE. ON THE OTHER HAND, ANBAR _ET _AL. (1965) REPORTED THE RESULTS OF EXPERIMENTS WHICH INDICATED DISSOCIATION OF THE CALORIGENIC EFFECT OF THYROID HORMONE FROM DEIODINATION. THEY FOUND THAT FOLLOWING THE INJECTION OF THYROXINE TO RATS, BASAL METABOLIC RATE WAS STIMULATED PRIOR TO DEIODINATION OF THYROXINE AND THAT BASAL METABOLIC RATE STIMULATION HAD LARGELY DIMINISHED BY THE TIME PEAK DEIODINATION ACTIVITY WAS OBSERVED. |F INCREASED DEIODINATION RATE IS A CONSEQUENCE OF ACCELERATED RESPIRATION RATHER THAN THE CAUSE, THEN SOME OTHER EXPLANATION FOR THE EFFECT OF VITAMIN E IN REDUCING THYROID ACTIVITY IS NECESSARY. |T IS INTERESTING THAT 0.25$ EQ EXERTED A GREAT EFFECT ON THE THYROIDAL RESPONSE (TABLES 2 AND 6, FIGURES 1 AND 2), BUT THE REVERSE 20 EFFECT OF THAT WAS OBTAINED WITH EXCESS VITAMIN E, ALTHOUGH BOTH VITAMIN E AND EQ ARE ANTIOXIDANTS. AT A LEVEL OF 0.25$ EQ IS TOXIC (MARCH ET AL., 1968). THE MECHANISM BY WHICH EQ INCREASES THE GOITROGENICITY OF THIOURACIL IS OBSCURE. EXPERIMENT 1c ,131 THIS EXPERIMENT WAS DESIGNED TO STUDY THE RATES OF I UPTAKE AND RELEASE BY THE THYROID GLAND IN CHICKS IN RESPONSE TO FEEDING DIETS CONTAINING LOW AND EXCESSIVE LEVELS OF VITAMIN E. MATERIALS AND METHODS THE COMPOSITION OF THE BASAL DIET AND THE LEVEL OF VITAMIN E SUPPLEMENTATION WERE THE SAME AS IN EXPERIMENT 1B. TWO HUNDRED DAY-OLD WHITE LEGHORN MALE CHICKS WERE DISTRIBUTED AT RANDOM INTO TWO GROUPS OF ONE HUNDRED. ONE GROUP WAS FED THE BASAL DIET, AND THE OTHER GROUP THE VITAMIN E SUPPLEMENTED DIET. AT 23 DAYS OF AGE, ALL THE BIRDS OF THE TWO GROUPS WERE WEIGHED INDIVIDUALLY AND 40 CHICKS FROM EACH GROUP WERE SELECTED SYSTEMATICALLY AS IN EXPERIMENT 1A. ROOM TEMPERATURE WAS THERMOSTATICALLY CONTROLLED AT 22-23°C. THE 40 CHICKS OF EACH GROUP WERE DISTRIBUTED SYSTEMICALLY INTO 5 LOTS OF 8 CHICKS EACH. THE CHICKS WERE 1 31 INJECTED INTRAVENOUSLY WITH 0.225 JiCI OF I IN 0.45 ML OF PHYSIOLOGICAL SALINE SOLUTION. FlVE, 10, 24, 48 AND 96 HOURS AFTER INJECTION, 8 CHICKS FROM EACH GROUP WERE WEIGHED. THE CHICKS WERE THEN KILLED AND THEIR T J31 THYROIDS REMOVED AND WEIGHED. THE I RADIOACTIVITY OF EACH PAIR OF GLANDS WAS MEASURED WITH A DEEP-WELL SCINTILLATION DETECTOR. MEASUREMENTS 21 OF RADIOACTIVITY WERE CORRECTED FOR COINCIDENCE LOSS, BACKGROUND RADIATION AND RADIOACTIVE DECAY. THE CORRECTED COUNTS WERE EXPRESSED AS A PERCENTAGE OF THE INJECTED DOSE, THE RADIOACTIVITY OF WHICH WAS DETERMINED FROM A STANDARD. THE STANDARD WAS PREPARED BY DILUTING 5 ML. OF THE INJECTION SOLUTION INTO 500 ML. OF PHYSIOLOGICAL SALINE SOLUTION. RADIOACTIVITY WAS MEASURED IN A 1 ML. ALIQUOT OF THIS SOLUTION USING A DEEP-WELL SCINTILLATION DETECTOR. ,131 THE PERCENT I UPTAKE WAS EXPRESSED PER BIRD AND. PER MG. OF THYROID GLAND. RESULTS AND DISCUSSION THYROIDAL RADIOACTIVITY AT DIFFERENT TIMES AFTER ADMINISTRATION OF i 3i I ARE SHOWN IN TABLE 7 AND IN FIGURES 3 AND 4.. REGARDLESS OF THE METHOD OF EXPRESSING THYROIDAL RADIOACTIVITY (.WHETHER PER BIRD OR PER MG. OF THYROID GLAND) THE DATA INDICATE THAT THE ACTIVITY OF THE THYROID GLAND IS ,131 DEPRESSED IN BIRDS FED EXCESS VITAMIN E. THE LEVELS OF I IN THE THYROID GLAND OF CHICKS FED SUPPLEMENTARY VITAMIN E WERE LOWER WHEN MEASURED 5 AND .131 _ 10 HOURS AFTER I INJECTION THAN IN THE CONTROL CHICKS. THE DIFFERENCES BETWEEN THE CHICKS RECEIVING THE RESPECTIVE LEVELS OF VITAMIN E WERE STATISTICALLY SIGNIFICANT AT 5 AND 10 HOURS AFTER INJECTION WHEN CALCULATED ON THE BASIS OF RADIOIODINE UPTAKE PER BIRD AND WERE SIGNIFICANT AT 5, 10 AND 24 HOURS AFTER INJECTION WHEN CALCULATED ON THE BASIS OF RADIOIODINE UPTAKE PER MG. OF THYROID TISSUE. SUBSEQUENT TO 24 HOURS AFTER INJECTION ,131 TIME, THERE WAS A RAPID DECLINE IN THYROIDAL I IN THE CONTROL BIRDS WHEREAS THE LOSS OF RADIOACTIVITY FROM THE THYROID OF THE BIRDS FED EXCESS VITAMIN E WAS NOT SIGNIFICANT. THIS EXPERIMENT CONFIRMS THE PREVIOUS 131 FINDINGS (VALENTI AND BOTTARE L L I , 1965; DURAND _E_T _AL,., 1966) THAT I UPTAKE BY THE THYROID GLAND WAS REDUCED IN MAMMALS GIVEN A SUBCUTANEOUS INJECTION OF AN EXCESSIVE LEVEL OF VITAMIN E. |N THIS EXPERIMENT, 0.5$ IODIZED SALT WAS ADDED TO THE DIET TO ENABLE NORMAL IODINE METABOLISM IN THE THYROID GLAND. KOBAYASHI AND GORBMAN (1960) AND ROSENBERG _ET _AL. (1964) HAVE DEMONSTRATED IN RATS AND CHICKS THAT THE 22 .131 PERCENT OF INJECTED I IN THYROID GLAND * CO m i "n — "n H m PI o -l t~ m O SD "n I O n 33 X Z o m CO > CO V n o o n X -l — > o -< co • < n: — o —i c > 33 2 CO z > m H n o 33 z -1 —_> I CO -< —1 33 O — — z C-> n (- o H o O o z z m ,—. CO c "0 H > > Z o 33 m r~ m > CO m £2 t/2 25 ,131 TIME OF PEAK I UPTAKE WAS INFLUENCED BY THE LEVEL OF DIETARY IODINE. KOBAYASHI AND GORBMAN (I960) HAVE SHOWN THAT IN CHICKS FED A LOW IODINE DIET THYROIDAL RADIOACTIVITY FELL RAPIDLY AFTER REACHING A PEAK AT 6 HOURS. ROSENBERG _ET _AL. (1964) FOUND THAT IN COCKERELS FED AN IODIDE SUPPLEMENTED 131 DIET, AVERAGE MAXIMAL I UPTAKE OF 13$ OF THE INJECTED DOSE WAS REACHED 12 HOURS AFTER INJECTION AND A VERY LONG RETENTION OF THYROIDAL RADIOIODIDE OCCURED. THE RESULTS OF THIS EXPERIMENT, WHICH SHOWED THAT BOTH THE RATES OF IODINE UPTAKE AND OF IODINE RELEASE BY THE THYROID GLAND ARE DEPRESSED IN CHICKS FED EXCESS VITAMIN E, SUBSTANTIATE THE CONCLUSIONS OF EXPERIMENTS 1A, AND B, WHEREAS IN EXPERIMENT 1, HOWEVER, THE DATA INDICATED THAT TSH SECRETION WAS REDUCED IN HYPERVITAMINOSIS E, EXPERIMENT 1C. PROVIDES 'D I RECT EVIDENCE THAT THYROID SECRET I ON RATE IS LESS IN EXCESS VITAMIN E FED CHICKS. • EXPERIMENT I I RESPIRATION RATE OF MUSCLE MITOCHONDRIA FROM CHICKS FED EXCESS DIETARY VITAMIN E THE EXPERIMENT WAS DESIGNED TO STUDY THE OXYGEN CONSUMPTION OF MUSCLE MITOCHONDRIA FROM NORMAL AND THIOURAC IL-TREATED CHICKS FED NORMAL AND EXCESS LEVELS OF VITAMIN E. MATERIALS AND METHODS SIXTY DAY-OLD MALE WHITE LEGHORN CHICKS WERE DISTRIBUTED INTO TWO GROUPS IN ELECTRICALLY HEATED BATTERY BROODERS. THE CONTROLS RECEIVED THE BASAL DIET WHILE THE OTHER GROUP WAS FED THE BASAL DIET SUPPLEMENTED WITH 2200 |.U. OF DL-OL-TOCOPHERYL ACETATE PER KG. AFTER 40 DAYS, 15 CHICKS OF EACH GROUP WERE FED THE RESPECTIVE DIET WITH THE ADDITION OF 0.1$ OF THIOURACIL. FIFTEEN DAYS LATER, THE CHICKS (55 DAYS OLD) WERE KILLED AND THE PECTORAL MUSCLE EXCISED IMMEDIATELY FOR THE ISOLATION OF MITOCHONDRIA. PREPARATION OF MITOCHONDRIA APPROXIMATELY 15 GM OF MUSCLE FROM EACH CHICK WAS IMMERSED IN ICE-CHILLED 0.15M KC1 SOLUTION. THE TISSUE WAS THEN HOMOGENIZED WITH 100 ML OF THE ICE-CHILLED CHAPPELL- PERRY TRIS-KC1 MEDIUM (1954), USING A VLRTIS HOMOGENIZER AT A SETTING OF 70 FOR 45 SECONDS. MYOFIBRILS, NUCLEI AND UNBROKEN CELLS WERE REMOVED BY TWO SUCCESSIVE CENTRIFUGATIONS AT 600 X G FOR 10 MINUTES AT 0-4°C (AZ Z ONE E_T _AL . , 1961). THE FINAL 26 27 SUPERNATANT FLUID WAS THEN CENTRIFUGED AT 8500 X G FOR 10 MINUTES AT 0-4°C. THE MITOCHONDRIA WERE WASHED TWICE BY RESUSPENSION IN THE ISOLATION MEDIUM AND RECENTRI FUGED AT 8500 X G. THE MITOCHONDRIAL PELLET WAS RINSED WITH 0.15M KC1 AT A CONCENTRATION OF APPROXIMATELY 3 MG. OF MITOCHONDRIA PER ML. THE PROTEIN CONTENT OF THE MITOCHONDRIAL SUSPENSION WAS DETERMINED BY THE BIURET METHOD (G0RNAL JET _AL . , 1949) AFTER CLEARING THE SUSPENSION WITH 0.5$ SODIUM DEOXYCHOLATE. CRYSTALLINE BOVINE SERUM ALBUMIN WAS USED AS A STANDARD TO PLOT A CALIBRATION CURVE FOR PROTEIN ESTIMATIONS. OXYGEN UPTAKE BY MITOCHONDRIA RESPIRATION OF MITOCHONDRIA WAS MEASURED BY THE WARBURG MANOMETRIC TECHNIQUE (UMBR I E T _E_T _A_L ., 1959). THE MEDIUM USED WAS THAT OF AZZONE (1961 ) WITH SOME MOD I F1 CAT I ON. EACH VESSEL CONTAINED 25 MM TRIS BUFFER (PH 7.5), 50 MM KC1, 6 MM MGCL2, 20 MM PI (PH 7.5), 0.01 MM O(-KETO-GLUTARATE, 0.02 MM CYTOCHROME C, 1 MM ATP (oI-NA-SALT), 30 MM GLUCOSE AND 0.33 MG. YEAST HEXOKINASE (SlGMA TYPE V). THE ADDITION OF GLUCOSE, HE XOKINASE AND CYTOCHROME C CAN STIMULATE THE RESPIRATION IN THE MEDIUM DURING INCUBATION PERIOD (HEDMAN, 1965). ATP WAS USED FOR PREVENTION OF MITOCHONDRIAL SWELLING (CHAPPEL AND PERRY, 1954). THE TOTAL LIQUID VOLUME WAS 3 ML PER WARBURG VESSEL. ALL FLASKS WERE EQUILIBRATED AT 30°C FOR 5 MINUTES IMMEDIATELY AFTER ADDITION OF 1 ML OF THE MITOCHONDRIA TO THE REACTION MIXTURE. INCUBATION WAS AT 30°C WITH OXYGEN AS THE GAS PHASE FOR A PERIOD OF 2 HOURS. ALL DETERMINATIONS WERE PERFORMED IN DUPLICATE OR TRIPLICATE. THE OXYGEN UPTAKE OF THE MITOCHONDRIA WAS MEASURED AT 30 MINUTE INTERVALS FOR TWO HOURS. OXYGEN UPTAKE IS EXPRESSED AS UL/MG OF MITOCHONDRIAL PROTEIN/HOUR. ELECTRON MICROSCOPIC STUDY OF MUSCLE MITOCHONDRIA A SMALL SAMPLE OF A PELLET OF THE FRESH ISOLATED MITOCHONDRIA WAS TRANSFERRED INTO A MICROCENTRIFUGE TUBE ACCORDING TO THE METHOD OF MALAMED (1963) AND THEN FIXED IN \FO OS 0^ WITH A V E R ON A L- AC E T A TE ISOTONIC BUFFER AT PH 7.4 FOR 2 HOURS AT 4°C (KAY, 1966). THE SLURRY OF MITOCHONDRIA WAS REAGGREGATED INTO A MANAGEABLE PELLET IN 5 MINUTES OF CE N TRI FUGAT ION. AFTER DEHYDRATION IN A GRADED ALCOHOL SERIES AND FINALLY IN PROPYLENE OXIDE, THE SAMPLES WERE EMBEDDED IN EPOXY 812 (LUFT, 1961). ULTRATHIN SECT I ONS WERE MADE WITH MT-2 MICROTOME AND .MOUNTED ON CARBONED-COLLOD I ON-COATED 200 MESH COPPER GRIDS. DOUBLE STAINING WAS USED WITH URANYL ACETATE FOR 20 MINUTES (STEMPAK AND WARD, 1964) FOLLOWED BY LEAD CITRATE FOR 15 MINUTES (REYNOLDS, 1963). THE STAINED SECTIONS WERE EXAMINED WITH A HLTACHI-78 ELECTRON MICROSCOPE. RESULTS AND DISCUSSION THE. RESULTS SHOW A SIGNIFICANT DECREASE IN THE OXYGEN CONSUMPTION OF THE MUSCLE MITOCHONDRIA OF THE CHICKS FED EXCESSIVE LEVEL OF VITAMIN E, COMPARED WITH THE CHICKS FED THE CONTROL DIET (TABLE 8). THE DEPRESSION IN RESPIRATION DUE TO EXCESS VITAMIN E WAS SIGNIFICANT EVEN WHEN METABOLIC RATE HAD BEEN REDUCED BY THE FEEDING OF THIOURACIL. IT SHOULD BE NOTED THAT THE LEVEL OF VITAMIN E SUPPLEMENTATION IN.THIS EXPERIMENT WAS 10 TIMES THAT USED IN EXPERIMENT 1. BECAUSE TISSUE RESPIRATION RATE IS KNOWN TO BE ACCELERATED IN VITAMIN E DEFICIENCY, IT MAY BE DEDUCED THAT THE RESPONSE TO EXCESS VITAMIN E WILL BE RELATIVE TO THE LEVEL ADMINISTERED AND THAT A DEPRESSION IN RESPIRATION RATE WAS LIKEWISE ASSOCIATED WITH THE REDUCTION IN THYROID ACTIVITY WITH THE LEVEL OF VITAMIN E FED IN EXPERIMENT 1. A POSITIVE ION, I OR I RELEASED DURING THYROXINE DEGRADATION INTERACTS WITH THE PHOSPHOLIPIDS OF CELLULAR MEMBRANE, RESULTING IN A LOWERING OF THE ELECTRICAL RESISTANCE OF THE MEMBRANE (FLNKELSTEIN AND CASS, 1968J ROSENBERG AND JENDRASIAK, 1968). THIS LOWERED ELECTRICAL RESISTANCE, WHICH IS POSTULATED TO BE THE PRIMARY ACTION OF THYROXINE, MAY BE THE RESULT OF EITHER INCREASED IONIC CONDUCTIVITY, OR INCREASED ELECTRONIC CONDUCTIVITY, AND MAY LOWER THE RESISTANCE EITHER TRANSVERSE OR PARALLEL TO THE PLANE OF THE MEMBRANE. MITOCHONDRIA TRANSFER ELECTRONS FROM SUBSTRATES SUCH AS NADH OR SUCCINATE TO MOLECULAR OXYGEN ALONG THE CYTOCHROME CHAIN ASSOCIATED WITH THE MITOCHONDRIAL. MEMBRANE (GREEN ET _AL. , 1964). |T HAS BEEN SHOWN 29 30 THAT PHOSPHOLIPID FOR THE ACTIVITY OF THE ELECTRON TRANSPORTING SUBMITOCHONDRIAL FRACTIONS I, II, III AND IV WERE REQUIRED IN THE MEMBRANE OF MITOCHONDRIA ( B R I E R L Y _ET _AL . , 1962; BRIERLY AND MEROLA, 1962; FLEISCHER AND FLEISCHER, '1964). GRUENSTEIN (1970) SUGGESTED THAT, BY INC RE AS ING THE CONDUCTIVITY OF THE PHOSPHOLIPIDS, THYROXINE MAY EITHER FACILITATE THE TRANSFER OF ELECTRONS FROM CYTOCHROME TO CYTOCHROME, OR EVEN TO SOME EXTENT "SHORT CIRCUIT" PARTS OF THE ELECTRON TRANSPORT CHAIN, THUS CAUSING MORE RAPID REDUCTION OF OXYGEN AND PART IAL UNCOUPL I NG OF PHOSPHORYLATION. THIS ACTION IS IN AGREEMENT WITH THE OBSERVED INCREASED OXYGEN CONSUMPTION EFFECT OF THYROXINE ( TA T A _ET _AL . , 1 963) . EXCESS VITAMIN E MAY DEPRESS THE D EI 00 I NAT I ON OF THYROXINE (GALTON AND HLNGBAR, 1965). WITH A LESSER AMOUNT OF |+ OR |° RELEASED IN MEMBRANE OF MUSCLE MITCHONDRIA, CONDUCTIVITY OF THE PHOSPHOLIPID WILL BE LOWER, PHOSPHORYLATI ON .RATE WILL BE REDUCED AND OXYGEN UPTAKE IS DECREASED. IT IS CONCLUDED THAT EXCESS VITAMIN E NOT ONLY DEPRESSED THE ACTIVITY OF THYROID GLAND BUT ALSO REDUCE THE RESPIRATION RATE OF MUSCLE MITOCHONDRI A. THE GROWTH RATE OF THE CHICKS FED THE EXCESS LEVEL OF VITAMIN E IN THIS EXPERIMENT WAS MARKEDLY LOWER THAN THAT OF THE CONTROL CHICKS. |T IS.SUGGESTED THAT THE CHICK MAY MAINTAIN NORMAL RESPIRATION RATE WHEN FED EXCESS VITAMIN E UP TO A CERTAIN LEVEL THROUGH A REDUCTION IN THYROID ACTIVITY (SEE DISCUSSION OF EXPERIMENT 1). WITH A LARGE EXCESS OF VITAMIN E, HOWEVER, NO FURTHER COMPENSATION IS POSSIBLE AND RESPIRATION RATE IS DEPRESSED BELOW NORMAL. FROM THE ELECTRON MICROGRAPHS, SWOLLEN MITOCHONDRIA WERE CLEARLY 31 SEEN IN PREPARATIONS FROM BOTH CONTROL CHICKS AND CHICKS FED AN EXCESS OF VITAMIN E (FIGURES 5 AND 6). SOME SPECIMENS, HOWEVER, SHOWED NORMAL INTACT MITOCHONDRIA (FIGURE 7). THE SWOLLEN APPEARANCE OF THE MITOCHONDRIA MAY BE DUE TO THE SEVERAL WASHINGS WITH CHAPPEL AND PERRY TRIS-KCL MEDIUM. TABLE 8 INDICATES THAT THE RESPIRATION RATE OF THE MITOCHONDRIA IN THE PERIOD OF 0-30 MINUTES AND OF 30-60 MINUTES ARE NEARLY EQUAL. ORDINARILY IT IS REPORTED THAT RESPIRATION RATE DECLINES IN SUCCESSIVE 30 MINUTE PERIODS. IT APPEARS THAT THE MITOCHONDRIA MAY BE SWELLING CONTINUOUSLY DURING THE INCUBATION PERIOD OF 0-30 MINUTES UNTIL A CERTAIN AMOUNT OF ATP IS FORMED. THE SWELLING OF MUSCLE MITOCHONDRIA CAN BE REVERSED BY THE PRESENCE OF*LOW CONCENTRATIONS OF ATP (CHAPPEL AND PERRY, 1954) AND THUS THE MITOCHONDRIA UNDERGO A NORMAL RESPIRATION IN THE PERIOD OF 30-60 MINUTES. THE ELECTRON MICROGRAPH ALSO SHOWSTHAT THE MITOCHONDRIA PREPARATION WAS CONTAMINATED WITH SMALL PARTICLES WHICH MIGHT BE SUBMI TOCHONDRI A. |T APPEARS THAT THE METHOD OF PREPARATION OF MITOCHONDRIA USING THE FRACTION OBTAINED AT 8500 X G IS NOT A GOOD METHOD FOR OBTAINING INTACT MITOCHONDRIA FOR RESPIRATION STUDIES ALTHOUGH THE MITOCHONDRIAL YIELD IS GOOD. HEDMAN (1965) REPORTED THAT THE 3500 X G MITOCHONDRIAL FRACTION REPRESENTED RELATIVELY INTACT MITOCHONDRIA BUT THE MITOCHONDRIAL YIELD WAS REDUCED BY ABOUT ONE THIRD. |N THE PRESENT EXPERIMENT, TWO ADVERSE EFFECTS OF HYPERV I TAM INOSIS E WERE OBSERVED VIZ. BONE FRAGILITY AND AN INCREASE IN PROTHROMBIN TIME. 32 FIG. 5. ELECTRON MICROGRAPH OF BREAST MUSCLE MITOCHONDRIA FROM A CHICK FED A NORMAL LEVEL OF VITAMIN E. THE CONCENTRATION OF M I TOCHONDR I A WAS LOW AND THE SUSPENSION CONTAMINATED WITH SUBMITOCHONDRI A. THE SWOLLEN MITOCHONDRIA WERE CLEARLY SEEN. X 30,000. 33 FIG. 6. ELECTRON MICROGRAPH or A SECTION OF MUSCLE MITOCHONDRIA FROM A CHICK FED EXCESS VITAMIN E. THE SWOLLEN MITOCHONDRIA WERE CLEARLY SEEN AND THE SUSPENSION CONTAMINATED WITH SUBMITOCHONDRI A x 30,000. FIG. 7. ELECTRON MICROGRAPH or A SECTION or BREAST MUSCLE MITOCHONDRIA r ROM A CHICK FED A NORMAL LEVEL OF VITAMIN E SHOWING INTACT MITOCHONDRIA. NO SWELLING PHENOMENON WAS SEEN BUT THE MITOCHONDRIAL SUSPENSION WAS CONTAMINATED WITH SUBMITOCHONDRI A x 36,000. EXPERIMENT III SYMPTOMS or HYPERVITAMINOSIs E IN CHICKS THIS EXPERIMENT WAS DESIGNED TO INVESTIGATE THE EFFECTS OF EXCESS VITAMIN E IN CHICKS FED DIETS DEFICIENT AND ADEQUATE IN CALCIUM. SLNCE THERE IS NO INFORMATION AVAILABLE REGARDING THE EFFECTS OF LONG TERM ADMINISTRATION OF EXCESS VITAMIN E TO CHICKS, THE EXPERIMENT WAS CONDUCTED WITH CHICKS FED THE DIFFERENT DIETS FROM HATCH - TO 50 DAYS OF AGE. IN EXPERIMENT II, TWO ADVERSE EFFECTS OF HYPERVITAMINOSIS E WERE OBSERVED VIZ. BONE FRAGILITY AND AN INCREASE IN PROTHROMBIN TIME. THE DIETS FED IN THIS EXPERIMENT WERE ACCIDENTLY FORMULATED WITHOUT ADEQUATE CALCIUM SUPPLEMENTATION. THERE WAS, THEREFORE, THE POSSIBILITY OF AN INTERRELATIONSHIP BETWEEN THE CALCIUM DEFICIENCY AND EXCESS VITAMIN E IN THE RESPONSE OF THE CHICKS IN THIS EXPERIMENT. A SEARCH OF THE LITERATURE FOR RELATED OBSERVATIONS REVEALED ONE REPORT THAT HIGH DIETARY LEVELS OF EITHER VITAMIN A OR VITAMIN E DEPRESSED PROTHROMBIN LEVEL OF THE BLOOD IN THE RAT (MELLETE AND LEONE, 1960). NO REPORT OF ANY EFFECT OF EXCESS VITAMIN E ON BONE CALCIFICATION WAS' FOUND. MATERIALS AND METHODS DAY-OLD WHITE LEGHORN MALE CHICKS WERE WING-BANDED AND DISTRIBUTED AT RANDOM INTO 8 LOTS OF 25 CHICKS EACH IN COMPARTMENTS OF ELECTRICALLY HEATED BATTERY BROODERS. THE FOUR DIFFERENT DIETS USED IN THE EXPERIMENT 35 36 ARE SHOWN IN TABLE 9. EACH DIET WAS FED TO DUPLICATE LOTS OF CHICKS. ROOM TEMPERATURE WAS THERMOSTATICALLY CONTROLLED AT 22-23°C. THE FEED AND WATER WERE SUPPLIED AD LIBITUM. AFTER A PERIOD OF 30 DAYS, BLOOD SAMPLES (1 ML PER CHICK) OF 3 CHICKS FROM EACH LOT WERE DRAWN AT RANDOM FROM THE WING VEIN BY USING TUBERCULIN SYRINGE. BLOOD SAMPLES . C0NTA INED SODIUM OXALATE OR HEPARIN FOR THE DIFFERENT HEMATOLOGICAL DETERMINATIONS. PROTHROMBIN TIMES, HEMATOCRIT VALUES AND RETICULOCYTE COUNTS WERE DETERMINED ON THE BLOOD SAMPLES. PROTHROMBIN TIME THE MEASUREMENT OF PROTHROMBIN TIME IS USED AS AN IMPORTANT SCREENING TEST FOR THE DEFICIENCY OF THEVARIOUS FACTORS OF THE PROTHROMBIN COMPLEX SUCH AS PROTHROMBIN, FACTORS V, VII AND X (QUICK, 1961). NINE-TENTH ML OF BLOOD WAS DRAWN IN A SYRINGE' CONTAINING 0.1 ML OF SODIUM OXALATE AND MIXED WELL. PROTHROMBIN TIME WAS ESTIMATED BY THE QUICK ONE STAGE METHOD (QUICK, 1936) WITH MODIFICATION BY ALMQU1ST (1941). HEMATOCRIT VALUE THE DETERMINATION OF HEMATOCRIT VALUE (OR PACKED CELL VOLUME) IS USED AS A SIMPLE SCREENING .TEST FOR ANEMIA. THE MICROMETHOD WAS APPLIED, ML C RO- HE.MATOC R I T TUBES (1.1-1.2 MM. IN I.D., 75 MM IN 1 LENGTH) WERE USED FOR THE HEMATOCRIT DETERMINATIONS. THE HEPARINIZED BLOOD SAMPLE WAS ALLOWED TO ENTER THE TUBE BY CAPILLARITY. THE TUBE WAS THEN SEALED WITH PLASTICINE. AFTER CENTRIFUGATION FOR 3 MINUTES, USING A INTERNATIONAL MICRO-CAPILLARY CENTRIFUGE, MODEL M.B. THE HEMATOCRIT VALUE WAS MEASURED USING A READING DEVICE. 37 RETICULOCYTE COUNTS THE RETICULOCYTE STAIN TECHNIQUE (COATES AND MARCH, 1966) WAS USED FOR RETICULOCYTE COUNTS. AT 30 DAYS OF AGE, 4 CHICKS FROM EACH LOT WERE WEIGHED AND KILLED. THE LEFT TIBIA OF EACH CHICK WAS TAKEN FOR ASH ANALYSIS. BONE ASH, EXPRESSED ON A DRY FAT-FREE BASIS, WAS USED TO SHOW THE DEGREE OF BONE CALCIFICATION IN THE LEFT TIBIA ACCORDING TO THE A.O.A.C. PROCEDURE (1965), USED IN THE CHICK ASSAY FOR VITAMIN D. THE RIGHT TIBI AS WERE CUT AND THE APPEARANCE OF THE MARROW NOTED. THESE OBSERVATIONS WERE REPEATED^ ON THE REMAINING CHICKS OF 40 AND 50 DAYS OF AGE. INJECTION OF VITAMIN K AT THE CONCLUSION OF THE EXPERIMENT, THREE BIRDS FED EXCESS VITAMIN E WERE INJECTED INTRAMUSCULARLY WITH MENADIONE (5 MG./KG. OF BODY WEIGHT) IN OIL SOLUTION. TWENTY-FOUR HOURS AFTER INJECTION, PROTHROMBIN TIME WAS DETERMI NED. PROTHROMBIN TIMES WERE NOT MEASURED AT 50 DAYS OF AGE. RESULTS PROTHROMBIN TIME THE MEAN PROTHROMBIN TIME OF THE CHICKS FROM EACH DIETARY TREATMENT AND THE STATISTICAL ANALYSIS OF THE RESULTS ARE SHOWN IN TABLE 10. THE ADDITION OF EXCESS VITAMIN E TO EITHER THE BASAL DIET OR TO THE BASAL DIET SUPPLEMENTED WITH ADDITIONAL CALCIUM SIGNIFICANTLY (P$0.01) PROLONGED PROTHROMBIN TIME OF BLOOD FROM THE CHICKS. AT 30 DAYS OF AGE, THERE WAS AN EFFECT OF DIETARY CALCIUM LEVEL ON PROTHROMBIN TIME. AT 40 DAYS OF AGE o EXCESS VITAMIN E LENGTHENED PROTHROMBIN TIME TO A GREATER DEGREE WHEN THE DIET WAS DEFICIENT IN CALCIUM. COMPARISON SHOULD NOT BE MADE BETWEEN THE ABSOLUTE PROTHROMBIN TIMES MEASURED AT 30 AND 40 DAYS OF AGE BECAUSE OF A POSSIBLE DIFFERENCE IN THE ACTIVITY OF THE THROMBOPLASTIN PREPARATION USED IN THE TWO SETS OF MEASUREMENTS. HEMATOCRIT VALUES THE HEMATOCRIT VALUES ARE ALSO SHOWN IN TABLE 10. EXCESS VITAMIN E RESULTED IN DECREASED HEMATOCRIT VALUES IN THE PRESENT EXPERIMENT. SUPPLEMENTATION OF THE BASAL DIET TO'SUPPLY ADEQUATE CALCIUM INCREASED SIGNIFICANTLY THE HEMATOCRIT VALUE OF BLOOD SAMPLES AT 30 DAYS OF AGE (P $0.01). BY 40 DAYS OF AGE THE DIFFERENCES IN HEMATOCRIT VALUES BETWEEN THE CHICKS FED THE CALCIUM DEFICIENT AND CALCIUM-ADEQUATE DIETS WERE NO LONGER SIGNIFICANT. AT 50 DAYS OF AGE, THE DIFFERENCES IN HEMATOCRIT VALUES OF BLOOD SAMPLES FROM THE CHICKS FED NORMAL AND EXCESS LEVELS OF 38 39 VITAMIN E, ALTHOUGH STATISTICALLY SIGNIFICANT (P^GO.OL) WERE SMALLER THAN THOSE OBSERVED WITH YOUNGER BIRDS. RETICULOCYTE COUNTS RETICULOCYTE COUNTS OF THE CHICKS FED THE BASAL DIET AND THE BASAL DIET SUPPLEMENTED WITH CALCIUM ARE SIMILAR (TABLE 10). SUPPLEMENTATION OF THE DIETS WITH EXCESS.VITAMIN E INCREASED RETICULOCYTE COUNTS SIGNIFI CANTLY (P 0.01) IN THE BLOOD FROM CHICKS FED DIETS OF EITHER CALCIUM LEVEL. IN THE BLOOD SAMPLE FROM CHICKS FED EXCESS VITAMIN E, MANY EARLY RETICULOCYTES WITH A HEAVILY STAINING RETICULUM WERE SEEN. THE CELLS OBSERVED IN MANY OF THE BLOOD SMEARS FROM THE CHICKS FED EXCESS VITAMIN E APPEARED TO BE ERYTHR0BLASTS (COMPARABLE TO FIGURES 9-13 OR 121-124, LUCAS AND JAMROZ, 1961). PHOTOGRAPHS OF REPRESENTATIVE CELLS ARE SHOWN IN FIGURES 8, 9 AND 10. BONE MARROW THE MARROW CAVITY OF THE TIBIA FROM BIRDS FED EXCESS VITAMIN E SHOWED RESORPTION OF ALL CALCIFIED TISSUE AND CONTAINED ONLY BLOODY MATERIAL. THE TYPICAL APPEARANCE IS SHOWN IN FIGURE 11 FROM A BIRD 50 DAYS OF AGE. ! BONE CALCIFICATION DATA RELATING TO THE CALCIFICATION OF BONE ARE SHOWN IN TABLE 12. IN GROUPS 3 AND 4 RECEIVING ON ADEQUATE LEVEL OF CALCIUM (1$ CALCIUM) NO SIGNIFICANT DIFFERENCE IN BONE ASH PERCENTAGE WAS DETECTED IN HYPER-VITAMINOSIS E. IN COMPARING GROUPS, 1 (NORMAL LEVEL OF VITAMIN E) AND 2 (HYPERVITAMINOSIS E), BOTH RECEIVING A LOW LEVEL OF CALCIUM FIG. 9. RETICULOCYTES FROM A 40-DAY OLD VITAMIN E x 1000. CHICK FED EXCESS 41 FlG. 10. ERYTHROBLASTS OBSERVED ON THE A 40-DAY OLD CHICK X 1000. BLOOD SMEAR FROM 42 E D FIG. 11. ABNORMAL BONE MARROW OF CHICKS FED EXCESS V ITAMIN E. A. NORMAL BONE MARROW FROM THE 50-DAY OLD CONTROL CHICK. B. SEVERE HYPOPLASTIC BONE MARROW FROM THE 50-DAY OLD CHICK FED EXCESS VITAMIN E. C. MEDIATE HYPOPLASTIC BONE MARROW FROM A 40-DAY OLD CHICK FED EXCESS VITAMIN E DIET. D. SLIGHTLY HYPOPLASTIC BONE MARROW FROM A 40-DAY OLD CHICK FED EXCESS VITAMIN E. 43 (0.59$ CALCIUM), THE ASH PERCENTAGE WAS SIGNIFICANTLY LOWER IN GROUP 2 IN PERIODS OF 30, 40 AND 50 DAYS. BECAUSE BONE MINERALIZATION INCREASED AS THE CHICK BECAME OLDER, AND HEAVIER AND BECAUSE EXCESS VITAMIN E DEPRESSED GROWTH, AT LEAST PART OF THE DIFFERENCE IN BONE ASH BETWEEN THE CHICKS FED THE NORMAL AND EXCESS LEVELS OF VITAMIN E MAY HAVE BEEN DUE TO THE DIFFERENCE IN BODY WEIGHT OF THE BIRDS OF THE TWO LEVELS OF VITAMIN E. PERCENT TIBIA ASH WAS ACCORDINGLY GRAPHED AGAINST BODY WEIGHT IN FIGURE 12B. FOR COMPARISON, A GRAPH WAS ALSO MADE OF PERCENT TIBIA ASH RELATIVE TO AGE OF THE BIRDS. USING THE VALUES FOR PERCENT ASH AT A BODY WEIGHT OF 300 GM., IT WILL BE SEEN THAT, WHEN THE DIET WAS DEFICIENT IN CALCIUM, AN EXCESS OF VITAMIN E DEPRESSED CALCIFICATION. INJECTI ON OF VI TAMIN K TWENTY-FOUR HOURS AFTER INJECTION OF MENADIONE, PROTHROMBIN TIME OF THE CHICKS FED EXCESS VITAMIN E WAS SHORTENED TO THAT OF CHICKS FED THE NORMAL LEVEL OF VITAMIN E (TABLE 11). DISCUSSION THE- PROLONGED PROTHROMBIN TIME IN THE CHICKS FED EXCESS VITAMIN E AND THE SHORTENING OF PROTHROMBIN TIME BY ADMINISTRATION OF MENADIONE WERE INDICATIVE OF VITAMIN K-DEFICIENCY (ALMQUIST AND ST.OKSTAD, 1936). THE PROLONGED PROTHROMBIN TIME IN CHICKS FED EXCESS VITAMIN E CONFIRMED THE OBSERVATION BY MELLETE AND LEONE (i960) IN RATS. THE MECHANISM BY WHICH HYPOPROTHROMBINEMI A IS CAUSED MAY BE INTERPRETED IN ONE OF THREE WAYS. ONE POSSIBLE MECHANISM IS THAT AN EXCESSIVE AMOUNT OF VITAMIN E OR ITS METABOLITES MAY ANTAGONIZE THE ACTION OF VITAMIN K. IN THE COURSE OF EARLIER STUDIES ON THE METABOLISM OF oC-TOCOPHERYL-5-METHYL-C IN ANIMALS, THREE LABELED METABOLITES, oC- TOC OPHE RyL-P- QU I NONE (CSALLANY _E_T _AL . , 1962), A DIMER (CSALLANY AND DRAPER, 1963) AND A TRIMER (D RAPE R _E_T _AL . , 1967) WERE ISOLATED FROM THE LIVER TISSUE. |T IS SUGGESTED THAT THE STRUCTURE OF THESE END METABOLITES, HOWEVER, ARE SIMILAR TO THAT OF VITAMIN K AND PROBABLY COMPETE WITH VITAMIN K FOR CERTAIN ENZYME SITES. THE SECOND POSSIBLE MECHANISM IS THAT AN EXCESSIVE AMOUNT OF VITAMIN E MAY INTERFERE WITH THE ABSORPTION OF FAT SOLUBLE VITAMINS, SUCH AS VITAMINS K AND D. THE THIRD IS THAT EXCESSIVE AMOUNTS OF VITAMIN E MAY RETARD THE SYNTHESIS OF VITAMIN K. EXCESS VITAMIN E DID NOT EXERT AN EFFECT ON THE BONE FORMATION IN CHICKS FED ADEQUATE CALCIUM DIET. WHEN FED A LOW CALCIUM DIET, PERCENT ASH IS SIGNIFICANTLY LOWER IN CHICKS FED EXCESS VITAMIN E THAN THAT IN THE CHICKS FED THE BASAL DIET, SHOWN IN TABLE 12 AND FIGURES 12A AND B. IT 44 PERCENT OF TIBIA ASH PERCENT OF TIBIA ASH CO r-i ro 00 O O ro -1 —1 o — I o CD pi > _ z > "n TO CO f- O X c O CO n -< o 70 z c pi o s: m pi > — O o — ^1 X < H pi Pi X . •t» —i o CD o o PI o co —^ CD co o D < -< — s: > <J1 pi o — — o z X -1 m O z CJ) -o o PI o X) o PI z H o •n CO 4* o JtD-4* co ro —* + 00 CO + CO CO > > > > PI CO PI CO CO X > > X > > n r" r o pi m CO + + CO .—- -—. co CO r-o O o < > > < c c -i o o > > > > 2 -— — Z z -n CD • ro > • o > —i -< — X CO CD PI — o > — z z > •n PI co r~ X X c "0 rn PI > z XI H n — PI 2 a PI — o z -n H pi pi XI X pi o z PI H CO co > CD < PI — CO • > Z rrl o z 13 PI XI r> z H O •n CO Q_ CO o -1^ o o CO o 4* O 4* CO ro —* + CO CO + CO CO > > > > PI co co PI CO CO X > > X > > (-) r" i- n p r- -PI pi CO + + co ,—. ,—^ co CO r- r o o O O < > > < s. -i o o > > > > 2 2 — — z z rn m 4* cn 46 APPEARS THAT EXCESS VITAMIN E MAY ALSO INTERFERE WITH THE ABSORPTION OF VITAMIN D OR SLOW DOWN THE BONE CALCIFICATION PROCESS. RET ICULOCYTOS1S HAS BEEN REPORTED TO OCCUR IN RESPONSE TO TOCOPHEROL TREATMENT OF VITAMIN E-DEFICIENT MONKEYS', HUMAN INFANTS AND CHILDREN (.FITCH, 1958; HASSAN rr AL,, 1966). MARCH ZJ_ AL. (1969) OBSERVED THAT RECICULOCYTOSIS OCCURED IN CHICKS FED EXCESS VITAMIN E (220 I.U. PER KG OF DIET) OR OTHER ANTIOXIDANTS AND ATTRIBUTED THE CONDITION TO RETARDATION OF THE MATURATION PROCESS OF THE RED BLOOD CELLS. MARC H _E_T _AL. ALSO REPORTED THAT ERYTHROCYTE TURNOVER WAS SLOWER IN SOME OF THE BIRDS FED EXCESS VITAMIN E AND THAT HEMATOCRIT VALUES WERE NOT REDUCED. IT IS I NTEREST I NG* I N THE PRESENT EXPERIMENT THAT THE LEVEL OF VITAMIN E (2200 I.U. PER KG OF DIET OR 100 TIMES THE NORMAL LEVEL) RESULTED IN THE DECREASED HEMATOCRIT VALUE,. RETICULOCYTOSIS AND ALTERATION OF BONE MARROW. ONE POSSIBLE MECHANISM IS THAT AN EXCESSIVE AMOUNT OF VITAMIN E NOT ONLY RETARDS THE MATURATION OF THE RETICULOCYTES IN THE BLOOD BUT ALSO SLOWS DOWN OR EVEN PREVENTS THE ERYTHROPOIES1S IN THE BONE MARROW, THEREFORE, RESULTING IN THE DECREASED HEMATOCRIT VALUE (ANEMIA). |N SOME SEVERE CASES, JELLY-LIKE BLOODY MATERIAL WAS OBSERVED IN THE MARROW CAVITY AND . ERYTHROBLASTS WERE PRESENT IN THE SMEARS OF THE CIRCULATING BLOOD INDICATING A RETARDATION OF ERYTHROPOIESIS. UNFORTUNATELY, MICROSCOPIC STUDIES HAVE NOT BEEN CARRIED OUT FOR BONE MARROW SMEAR. FURTHER STUDIES ARE REQUI RED. SUMMARY A STUDY WAS MADE OP THE EFFECTS OF FEEDING EXCESS VITAMIN E TO CHICKS. THREE GROUPS OF EXPERIMENTS WERE CONDUCTED TO INVESTIGATE (1) THYROIDAL RESPONSE TO EXCESS VITAMIN E, (2) RESPIRATION RATE OF MUSCLE MITOCHONDRIA FROM CHICKS FED EXCESS VITAMIN E, AND (3) SYMPTOMS OF HYPERVITAMI NO IS E. IN TWO SEPARATE EXPERIMENTS, CHICKS WERE FED NORMAL AND EXCESS LEVELS (220 I.U./KCJ. 0F DIET) OF VITAMIN E AND WERE SUBJECTED TO TEMPERATURES OF 14.5 AND 31.5°C. THE GOITROGENIC EFFECT OF THIOURACIL ON THE BIRDS IN THE DIFFERENT TREATMENTS WAS MEASURED. AT BOTH TEMPERATURES THE THIOURACIL-TREATED CHICKS FED EXCESS VITAMIN E EXHIBITED A LESSER ENLARGEMENT OF THE THYROID GLANDS THAN DID CHICKS RECEIVING A NORMAL LEVEL OF VITAMIN E. THIS FINDING INDICATES A REDUCTION IN THE SECRETION OF THYROID STIMULATING HORMONE (TSH) IN BIRDS. FED EXCESS VITAMIN E. THE LEVEL OF VITAMIN E WHICH WAS FED DID NOT AFFECT THE GROWTH RATE OR FEED CONSUMPTION AT EITHER TEMPERATURE. I T, THEREFORE, APPEARS THAT THE METABOLIC RATES OF THE CHICKS FED THE LOW AND EXCESS LEVELS OF VITAMIN E WERE SIMILAR DESPITE DIFFERENCES IN THYROID ACTIVITY AND THAT TISSUE RESPIRATION IN BIRDS FED EXCESS VITAMIN E CAN BE MAINTAINED WITH A REDUCED SUPPLY OR TURNOVER OF THYROID HORMONE. THE ACTIVITY OF THE THYROID GLAND ITSELF WAS STUDIED IN RESPONSE ,, 131^ TO EXCESS VITAMIN E IN ANOTHER EXPERIMENT. USING RADIOIODINE [I ) AS A 47 48 TRACER, IT WAS FOUND THAT THE RATES OF IODINE UPTAKE AND RELEASE BY THE THYROID GLAND WERE BOTH SLOWER IN BIRDS FED EXCESS VITAMIN E (220 I.U./KG. OF DIET) THAN IN CONTROL BIRDS. BECAUSE VITAMIN E ACTS AS A BIOLOGICAL ANTIOXIDANT, IT MAY, IN EXCESS AMOUNTS, DEPRESS THE RATE OF DEIODI NAT 1 ON OF THYROXINE IN THE PERIPHERAL TISSUE AT WHICH THYROID HORMONE IS REMOVED FROM THE CIRCULATION. AS A CONSEQUENCE, THE SECRETION OF TSH WOULD BE REDUCED. |N ORDER TO OBTAIN SOME INDICATION OF CAUSE AND EFFECT IN THE MECHANISM BY WHICH THE EXCESS VITAMIN E AFFECTS THYROTROPIC HORMONE SECRETION RATE AND THYROID ACTIVITY, THE RESPIRATION RATE OF MITOCHONDRIA ISOLATED FROM THE PECTORAL MUSCLE OF CHICKS WHICH HAD BEEN FED EXCESS VITAMIN E (2200 I.U./KG. OF DIET) WAS COMPARED WITH THAT OF THE CONTROL BIRDS. THE RESULTS SHOWED A SIGNIFICANT REDUCTION IN OXYGEN UPTAKE BY THE MUSCLE MITOCHONDRIA OF CHICKS FED THE EXCESS AMOUNT OF VITAMIN E. THE GROWTH RATE OF THE CHICKS FED THE EXCESS LEVEL OF VITAMIN E IN THIS EXPERIMENT WAS MARKEDLY LOWER THAN THAT OF THE CONTROL CHICKS. IT IS SUGGESTED THAT THE CHICK MAY MAINTAIN A NORMAL RESPIRATION RATE WHEN FED EXCESS VITAMIN E UP TO A CERTAIN LEVEL THROUGH A REDUCTION IN THYROID ACTIVITY. WlTH A LARGE EXCESS OF VITAMIN E, HOWEVER, NO FURTHER COMPENSAT I ON IS POSSIBLE AND RESPIRATION RATE IS DEPRESSED BE LOW NORMAL. EXCESS VITAMIN E CAUSED HYPOPROTHROMBINEMIA, INDICATIVE OF VITAMIN K-DEFICIENCY. OTHER SYMPTOMS NOTED WERE A REDUCTION IN HEMATOCRIT VALUES, RECTICULOCYTOSIS AND AN ABNORMALLY FLUID APPEARANCE OF THE BONE MARROW. BASED UPON OBSERVATIONS OF BONE CALCIFICATION THE CALCIUM REQUIREMENT APPEARED TO BE INCREASED IN THE PRESENCE OF EXCESS VITAMIN E. BIBLIOGRAPHY ALMQUIST, H. J., 1941. REPORT ON VITAMIN K: ASSAY BY CURATIVE BIOLOGICAL TEST. J. Assoc. OFF. ANAL. CHEM. 24:405-413. ALMQUIST, H. J. AND STOKSTAD, E. L. R., 1936. FACTORS INFLUENCING THE INCIDENCE OF DIETARY HEMORRHAGIC DISEASE IN CHICKS. J. NUTRITION 12:329-335. ANDREW, G. A., 1957. A FEW NOTIONS INVOLVED IN THE CLINICAL USE OF RADIOISOTOPES. ANN. INTERN. MED. 47:922-938. ANDIK, I., BALOGH, L. AND DONHOFFER, S., 1949. THE EFFECT OF THYROXINE IN THYROIDECTOMIZED RATS TREATED WITH METHYL THIOURACIL. EXPERIENTIA 5:249-250. ANBAR, A., INBAR, M. AND TATA, J. R., 1965. DISSOCIATION OF THE CALORIGENIC ACTION OF THYROID HORMONE FROM THE DEIODINATION OF L-THYROXINE IN vivo. ACTA ENDOCRINOL. 48:506-512. , 1965. VITAMINS AND OTHER NUTRIENTS. IN ASSOCIATION OF OFFICAL AGRICULTURE CHEMISTS. 10TH EDITION. ASTWOOD, E. B., 1945. CHEMOTHERAPY OF HYPERTHYROIDISM. HARVEY LECT. 40:195-235. AZZONE, G. F., EEG-OLOFSSON, 0., ERNSTER, L., LUFT, R. AND SZABOLCSI, G., 1961. STUDIES ON ISOLATED HUMAN SKELETAL MUSCLE MITOCHONDRIA. EXP. CELL RES. 22:415-436. BARKER, S. D., 1955. THE CIRCULATING THYROID HORMONE, PP. 74-89. BROOKHAVEN SYMPOSIA IN BIOLOGY #7, "THE THYROID" BROOKHAVEN NATIONAL LABORATORY, UPTON, NEW YORK. BARKER^ S. B., KIELY, C. E. AND LIPNER, H. J., 1949. METABOLIC EFFECTS OF THYROXINE INJECTED INTO NORMAL, THIOURACIL-TREATED AND THYROID-ECTOMIZED RATS. ENDOCRINOLOGY 45:624-629. BIRD, J. W., SCHOTTELIUS, D. D. AND SCHOTTELIUS, B. A., 1963. OXYGEN CONSUMPTION OF SOLEUS MUSCLES FROM NORMAL, VITAMIN E SUPPLEMENT AND VITAMIN E DEFICIENT GUINEA PIGS DURING REST AND STIMULATION. METABOLISM 12:570-575. 49 50 BRIERLEY, G. P. AND MEROLA, A. J., 1962. STUDIES OF THE ELECTRON-TRANSFER SYSTEM. XLVIII. PHOSPHOLIPID REQUIREMENTS IN CYTOCHROME OXIDASE. BIOCHIM. BIOPHYS. ACTA. 64:205-217. BRIERLEY, G. P., MEROLA, A. J. AND FLEISCHER, S., 1962. STUDIES OF THE ELECTRON-TRANSFER SYSTEM. LLIX. SITES OF PHOSPHOLIPID INVOLVEMENT IN THE ELECTRON-ELECTRON-TRANSFER CHAIN. BLOCHIM. BIOPHYS. ACTA. 64:218-228. BROWN-GRANT, K., 1957. THE "FEED-BACK" HYPOTHESIS OF THE CONTROL OF THYROID FUNCTION. CIBA FOUND. COLLOQ. ON ENDOCRINOLOGY 10:97-114. BUNYAN, J., GREEN, J., EDWIN, E. E. AND DIPLOCK, A. T., 1960. STUDIES ON VITAMIN E. 5. LIPID PEROXIDATION IN DIALURIC ACID-INDUCED HAEMOLYSIS OF VITAMIN E-DEFICIENT ERYTHROCYTES. BIOCHEM. J. 77:47-51. CALVERT, C. C, NESHEIM, M. C. AND SCOTT, M. L., 1962. EFFECTIVENESS OF SELENIUM IN PREVENTION OF NUTRITIONAL MUSCULAR DYSTROPHY IN THE CHICK. PROC. SOC. EXP. BIOL. MED. .108:16-18. CAPUTTO, R., MCCAY, P. B. AND CARPENTER, M. P., 1958. REQUIREMENT OF MN++ AND CA"1"*" FOR THE SYNTHESIS OF ASCORBIC ACID BY LIVER EXTRACTS OF ANIMALS DEPRIVED OF TOCOPHEROL. J. BIOL. CHEM. 233:1025-1029. CENTURY, B. AND HORWITT, M. K., 1965. BIOLOGICAL AVAILABILITY OF VARIOUS FORMS OF VITAMIN E WITH RESPECT TO DIFFERENT INDICES OF DEFICIENCY. FEDERATION PROC. 24:906-911. CENTURY, B. AND HORWITT, M. K., 1958. SOME FACTORS IN PRODUCTION OF ENCEPHALOMALAC I A IN THE VITAMIN E DEFICIENT CHICK. FEDERATION PROC. 17:473. CHALKAFF, I. L. AND TAUROG, A., 1949. THE FORMATION OF ORGANICALLY-BOUND IODINE COMPONENTS IN THE THYROID GLAND AND THEIR APPEARANCE IN PLASMA AS SHOWN BY THE USE OF RADIOACTIVE IODINE. ANN. N. Y. ACAD. Sci. 50:377-402. CHAPPELL, J. B. AND PERRY, S. V., 1954. BIOCHEMICAL AND OSMOTIC PROPERTIES OF SKELETAL MUSCLE MITOCHONDRIA. NATURE 173:1094-1095. CHAPMAN, D. AND LESLIE, R. B., 1970. STRUCTURE AND FUNCTION OF PHOSPHOLIPIDS IN MEMBRANES. |N MEMBRANES OF MITOCHONDRIA AND CHLOROPLASTS. PP. 91-126. EDITED BY RACKER, E., VAN NASTRAND REINHOLD CO., N. Y. CHEVILLE, N. F., 1966. THE PATHOLOGY OF VITAMIN E DEFICIENCY IN THE CHICK. PATH. VET. 3:208-225. CHRISTENSEN, F., DAM, H., GORTNER, R. A. AND SONDERGAAD, E., 1956. IN VITRO HEMOLYSIS OF ERYTHROCYTES WITH VITAMIN E DEFICIENT RATS AND CHICKS. ACTA PHYSIOL. SCANDINARICA 35:215-224. COATES, V. AND MARCH, B. E., 1966.' RECTICULOCYTE COUNTS IN THE CHICKS. POULTRY SCI. 45:1302-1303. CSALLANY, A. S. AND DRAPER, H. H. AND SHAN, S. N., 1962. CONVERSION OF 14 D-Ot-TOCOPHEROL-C TO TOC OPH E R YL-P-Q U I NON E IN VIVO. ARCH. BiocHEM. BIOPHYS. 98:142-145. CSALLANY, A. S. AND DRAPER, H. H., 1963. D I ME R I Z AT I ON OF CCr TOC OPH E ROL IN vivo. ARCH. BIOCHEM. BIOPHYS. 100:335-337. CUNNINGHAM, L. W., 1964. THE REACTION OF ft-LACTOGLOBULIN SULFENYL IODIDE WITH SEVERAL ANTITHYROID AGENTS. BIOCHEMISTRY (WASH.) 3:1629-1634. DAM, H., PRANGE, I. AND. SONDERGAAD, E., 1952. THE EFFECT OF CERTAIN SUBSTANCES ON VITAMIN A STORAGE IN THE LIVER OF THE RAT. ACTA PHARMACOL. TOXICOL. 8:23-29. DAM, H., 1957. INFLUENCE OF ANTIOXIDANTS AND REDOX SUBSTANCES ON SIGNS OF VITAMIN E DEFICIENCY. PHARMACOL. REV. 9:1-16. DEMPSEY, E. W. AND ASTWOOD, E. B., 1943. DETERMINATION OF THE RATE OF THYROID HORMONE SECRETION AT VARIOUS ENVIRONMENTAL TEMPERATURES. ENDOCRINOLOGY 32:509-518. DEMOLE, V., 1939. PHARMAKOLOGISCHES UBER VITAMIN E, VERTRAGLICHKE1T DES SYNTHETISCHEN D L- Oi-TOCOPHEROLS UND SEINES ACETATS. INTERN. Z. ViTAMiNFORSCH 8:338-341. DESAI, I. D., CALVERT, C. C, SCOTT, M. L. AND TAPPEL, A. L., 1964. PEROXIDATION AND LYSOSOMES IN NUTRITIONAL MUSCULAR DYSTROPHY OF CHICKS. PROC. SOC. EXP. BIOL. MED. 115:462-466. DESAI, I. D. AND SCOTT, M. L., 1965. MODE OF ACTION OF SELENIUM IN RELATION TO BIOLOGICAL ACTIVITY OF TOCOPHEROLS. ARCH. BLOCHEM. BIOPHYS. 110:309-315. DETWILER, T. C, GARRETT, R. H. AND NASON, A., 1966. EFFECTS OF DIGI TON I N AND TOCOPHEROL ON BOVINE HEART MUSCLE REDUCED D IPHOS-PHOPYRIDINE NUCLEOTIDE- AND SUCCI NATE-CYTOCHROME C REDUCTASE AND CYTOCHROME C OXIDASE. J. BIOL. CHEM. 241:1621-1631. 52 DRAPER, H. H., CSALLANY, A. S. AND CHIU, M., 1967. ISOLATION OF A TRIMER OF d-TOCOPHEROL FROM MAMMALIAN LIVER. LIPIDS 2:47-54. DURAND, N. , CZYHA, J. C, MORNEX, R. AND RERES, G., 1968. UPTAKE OF I 131 BY THYROID OF GOLDEN HAMSTER TREATED WITH VITAMIN E AND cL- TOCOPHERYL QU i NONE . C. R. Soc. B i OL . 162:888-890. CITED IN NUTRITION ABST. REV. 1969- 39:1113. ESCOBAR DEL REY, F. AND ESCOBAR, G. M., 1961. THE EFFECT OF PROPYL THIOURACIL, METHYLTHIOURACIL AND THIOURACIL ON PERIPHERAL METABOLISM OF L-THYROXINE IN THYROIDECTOM I ZED, L-THYROXINE MAINTAINED RATS. ENDOCRINOLOGY 69:456-465. ESCOBAR, G. M. AND ESCOBAR, DEL REY, F., 1962. INFLUENCE OF THIOUREA, POTASSIUM CHLORATE AND THIOCYANATE AND OF GRADED DOSES OF PROPYL THIOURACIL ON THYROID HORMONE METOBOLISM IN THYROIDECTOM I ZED RATS. ENDOCRINOLOGY 71:906-913. EVANS, H. M. AND BISHOP, K. S., 1922. ON THE RELATION BETWEEN FERTILITY AND NUTRITION. AMER. J. PHYSIOL. 63:396. EVANS, H. M., EMERSON, 0. H. AND EMERSON, G. A., 1936. THE ISOLATION FROM WHEAT GERM OIL OF AN ALCOHOL, (^-'TOCOPHEROL, HAVING THE PROPERTIES OF VITAMIN E. J. BIOL. CHEM. 133:319-332. FERNHOLZ, E., 1937. THE THERMAL DECOMPOSITION OF O(-TOCOPHEROL. J. AMER. CHEM. SOC. 59:1154-1155. FERNHOLZ, E., 1938. ON THE CONSTITUTION OF ©(-TOCOPHEROL. J. AMER. CHEM. SOC. 60:700-705. FLNKELSTEIN, A. AND CASS, A., 1968. PERMEABILITY AND ELECTRICAL PROPERTIES OF THIN LIPID MEMBRANES. J. GEN. PHYSIOL. 52:145S. FITCH, C. D., 1958. THE RED BLOOD CELL IN THE VITAMIN-E DEFICIENT MONKEY. AMER. J. CLIN. NUTRITION 21:51-56. FLOCK, E. V. AND BOLLMAN, J. L., 1962. THE EFFECT OF . THIOURACIL ON THE I METABOLISM OF L-THYROXINE. BIOCHEM. J. 84:621-626. FLEISCHER, A. C. AND FLEISCHER, B., 1964. A PHOSPHOLIPID REQUIREMENT FOR DPNH OXIDATION. FEDERATION PROC. 23:486. FLEISCHER, S. AND ROUSER, G., 1965. LIPIDS OF SUBCELLULAR PARTICLES. J. AMER. OIL CHEM. SOC. 42:588-607. FLEISCHER, S., ROUSCHER, B., CASU, A., KRITCHEVSKY, G., 1967. LIPID COMPOSITION OF MITOCHONDRIA FROM BOVINE HEART, LIVER, AND KIDNEY. J. LIPID. RES. 8:170-180. 53 FRANKLIN, A. L., LERNER, S. R. AND CHAIKOFF, I. L., 1944. THE EFFECT OF THIOURACIL ON THE FORMATION OF THYROXINE AND D I IODOTYROS INE BY THE THYROID GLAND OF THE RAT WITH RADIOACTIVE IODINE AS INDICATOR. ENDOCRINOLOGY 34:265-268. FRIEDMAN, I. AND MATTILL, H. A., 1941. THE OXYGEN CONSUMPTION OF SKELETAL MUSCLE FROM ANIMALS DEPRIVED OF VITAMIN E. AMER. J. PHYSIOL.. 131:595-600. -GALTON, V. A. AND INGBAR, S. H., 1965. EFFECTS OF VITAMIN DEFICIENCY ON THE J_N V I TRO AND J_N VIVO DEIODINATION OF THYROXINE IN THE RAT. ENDOCRINOLOGY 77:169-176. GORDON, A. H., GROSS, J., O'CONNOR, D. AND PITT-RIVERS, R., 1952. NATURE OF THE CIRCULATING THYROID HORMONE-PLASMA PROTEIN COMPLEX. NATURE 169:19-20. GORNALL, A. G., BARDAWILL, C. J. AND DAVID, M. M., 1949. DETERMINATION OF SERUM PROTEIN BY MEANS OF THE BIURET REACTION. J. BIOL. CHEM. 177:751-766. GREEN, J., 1962. INTERRELATIONSHIPS BETWEEN VITAMIN E AND OTHER VITAMINS AND THE UNIQUINONES. V I TAM |'NS AND HORMONES 20:485-491. GREEN, D. E., WHARTON, D. C, TZAGOLOFF, A., RIESKE, J. S. AND BRIERLEY, G. P., 1964. THE MITOCHONDRIAL ELECTRON-TRANSFER CHAIN, IN KING, T. E., MASON, H. S. AND MORRISON, M., (EDITOR), OXIDASES AND RELATED REDOX SYSTEMS. VOL. 2, WILEY, NEW YORK, PP. 1032-1101. GRIGOR'YEVA, V. A. AND SHCHUKINA, L. V., 1967. EFFECT OF E-AVITAMINOSIS ON THE OXIDATIVE PHOSPHORYLATION IN MITOCHONDRIA OF THE SKELETAL MUSCLES. UKR. BIOKHIM. ZH. 39(4):366-362. CITED IN BIOLOGICAL ABSTRACTS, 1968. 49:8460. GRUENSTEIN, E., 1970. A MOLECULAR MECHANISM OF ACTION OF THYROXINE: MODIFICATION OF MEMBRANE PHOSPHOLIPID BY IODINE. J. THEOR. BIOCHEM. 26:343-363. GUSTAFSSON, R., TATA, J. R., LINDBERG, 0. AND ERNSTER, L., 1965. THE RELATIONSHIP BETWEEN.THE STRUCTURE AND ACTIVITY OF RAT-SKELETAL MUSCLE MITOCHONDRIA AFTER THYROIDECTOMY AND THYROID HORMONE TREATMENT. J. CELL BIOL. 26:555-578. GUHA, A. AND ROELS, 0. A., 1965. THE INFLUENCE OF C^-TOCOPHEROL ON ARYLSULFATASE A AND B IN THE LIVER OF VITAMIN A DEFICIENT RATS. BIOCHIM. BIOPHYS. ACTA 111:364-374. 6 YORY, P. AND ROSE, C. S., 1948. EFFECT OF MORTALITY AND HEMOGLOBINURIA IN'RATS ALLOXAN. SCIENCE 108:716-718. DIETARY FACTORS ON EARLY FOLLOWING ADMINISTRATION OF HALMI, N. S. AND PITT-RIVERS, R., 1962. THE IODINE POOLS OF THE THYROID. ENDOCRINOLOGY 70:660-668. HALMI, N. S., 1961. THYROIDAL IODIDE TRANSPORT. 'VITAMINS AND H'ORMONES 19:133-163. HAMILTON, J. G. AND SOLEY, M. H., 1939. STUDIES IN IODINE METABOLISM BY USE OF A NEW RADIOACTIVE ISOTOPE OF IODINE. AMER. J. PHYSIOL. 127:557-572. HAMILTON, J. G. AND SOLEY, M., 1940. STUDIES IN IODINE METABOLISM OF THE THYROID GLAND IN SITU BY THE USE OF RADIOIODINE IN NORMAL SUBJECTS AND IN PATIENTS WITH VARIOUS TYPES OF GOITER. AMER. J. PHYSIOL. 131:135-143. HASSAN, H., HASHIM, S. A., VAN ITALLIE, T. B. AND SEHRELL, W. H., 1966. SYNDROME IN PREMATURE INFANTS ASSOCIATED WITH LOW PLASMA VITAMIN E LEVELS AND HIGH POLYUNSATURATED FATTY ACID DIET. AMER. J. CLIN. NUTRITION 19:147-157.' HEDMAN, H., 1965. PROPERTIES OF ISOLATED SKELETAL MUSCLE MITOCHONDRIA FROM RAT. Exp. CELL RES. 38:1-12. HENDRICH, C. E. AND TURNER, C. W., 1963. TIME RELATIONS IN THE ALTERATION OF THYROID GLAND FUNCTION IN FOWLS. POULTRY SCI. 42:1190-1195. HENDRICH, C. E. AND TURNER, C. W., 1964. ESTIMATION OF THYROID-STIMULATING HORMONE (TSH) SECRETION RATES OF NEW HAMPSHIRE FOWLS. PROC. SOC. EXP. BIOL. MED. 117:218-222. HENDRICH, C. E. AND TURNER, C. W., 1965. A COMPARISON OF THE EFFECTS OF ENVIRONMENTAL TEMPERA'TURE CHANGES AND 4.4 C COLD ON THE BIOLOGICAL HALF LIFE (rg) OF THYROXINE-I131 |N FOWLS. POULTRY Sci. 44:3-5. HENINGER, R. W., NEWCOMER, W. S. AND THAYER, R. H., 1960. THE EFFECT OF ELEVATED AMBIENT TEMPERATURE ON THE THYROXINE SECRETION RATE OF CHICKENS. POULTRY SCI. 39:1332-1337. HERRERA, E. F., ESCOBAR, DEL REY, F. AND ESCOBAR, G. M., 1963. EFFECT OF PROPYLTHIOURACIL ON THE IN VIVO DEIODINATION OF THYROXINE LABELED WITH I 131 |N DIFFERENT POSITIONS. ENDOCRINOLOGY 73:744-747. HERSHMAN, M. M. AND VAN MIDDLESWORTH, L., 1962. EFFECT OF ANTITHYROID COMPOUNDS ON THE DEIODINATION OF THYROXINE IN RATS. ENDOCRINOLOGY 71:94-100. HILLMAN, R. W., 1957. TOCOPHEROL EXCESS IN MAN. CREATINUIA ASSOCIATED WITH PROLONGED INGESTION. AMER. J. CLIN. NUTRITION. 5:597-600. HIMMEL, J. P. AND MELVILLE, P. S., 1951. RESPIRATION AND GLYCOLYSIS OF RABBIT MUSCLE IN VITAMIN E DEFICIENCY. J. BIOL. CHEM. 191:391-394. HOFFMANN, E. AND SHAFFNER, C. S., 1950. THYROID WEIGHT AND FUNCTION AS INFLUENCED BY ENVIRONMENTAL TEMPERATURE. POULTRY SCI. 29:365-376. HOGNESS, J. R., WONG, T. AND WILLIAMS, R. AFTER INJECTION OF RAD I 0-THYROXINE OR NORMAL RATS. METABOLISM 3:510 HORWITT, M. K., HARVEY, C. C. AND HARMAN, E. M., 1968. LIPIDS, oC-TOCOPHEROL AND ERYTHROCYTE HEMOLYSIS. VITAMINS AND HORMONES 26:487-499. HOUCHIN, 0. B., 1942. THE IN VITRO EFFECT OF O6-TOCOPHEROL AND ITS PHOSPHATE DER I VAT I VE- ON OXIDATION IN MUSCLE TISSUE. J. BLOL. CHEM. 146:313-321. HOUCHIN, 0. B. AND MATTILL, H. A., 1942A. THE OXYGEN CONSUMPTION, CREATINE AND CHLOIDE CONTENT OF MUSCLES FROM VITAMIN E-DEFICIENT ANIMALS AS INFLUENCED BY FEEDING -TOCOPHEROL. J. BIOL. CHEM. 146:301-307. HOUCHIN, 0. B. AND MATTILL, H. A., 1942B. THE INFLUENCE OF PARENTERAL ADMINISTRATION 0F c(-TOCOPHEROL PHOSPHATE ON THE METABOLIC PROCESSES IN DYSTROPHIC MUSCLE. J. BIOL. CHEM. 146:309-312. HOUCHIN, 0. B. AND MATTILL, H. A., 1942c. IN VITRO EFFECT OF (^-TOCOPHEROL PHOSPHATE ON OXYGEN CONSUMPTION OF MUSCLE FROM VITAMIN E DEFICIENT ANIMALS. PROC. SOC. EXP. BIOL. MED. 50:216-218. HUME, E. M., 1941. STANDARDIZATION OF VITAMIN E. NATURE 148:472-473. HUSTON, T. M., EDWARDS, H. M. AND WILLIAM, J. J., 1962. THE EFFECTS OF HIGH ENVIRONMENTAL TEMPERATURE ON THYROID SECRETION RATE OF DOMESTIC FOWL. POULTRY ScI. 41:640-644. JLROUSEK, L. AND CUNNINGHAM, L. M., 1968. STIMULATION OF THIOURACIL BINGING AND THE IODI NAT I ON SYSTEM IN BEEF THYROID MICROSOMES. BIOCHIM. BIOPHYS. ACTA 170:160-171. H., 1954. I EXCRETION INTO HYPERTHYROID, HYPOTHYROID -517. 56 JOINER, W. P. AND HUSTON, T. M., 1957. THE INFLUENCE OF HIGH ENVIRON MENTAL TEMPERATURE ON IMMATURE DOMESTIC FOWL. POULTRY Sci. 36:973-978. 1 31 JONES, S. L. AND VAN MIDDLESWORTH, L.; 1960. NORMAL I L-THYROXINE METABOLISM IN THE PRESENCE OF POTASSIUM PERCHLORATE AND INTERRUPTED BY PROPYLTHIOURACIL. ENDOCRINOLOGY 67:855-861. KAUNITZ, H. AND PAPPENHEIMER, A. M., 1942. OXYGEN CONSUMPTION IN VITAMIN E DEFICIENCY. AMER. J. PHYSIOL. 138:328-340. KAY, D. H., 1966. IN TECHNIQUES FOR ELECTRON MICROSCOPY, P. 172. F. A. DAVIS CO., PHILADELPHIA. KOBAYASHI, H. AND GORBMAN, A., 1960. RADIOIODINE UTILIZATION IN THE CHICKS. ENDOCRINOLOGY 66:795-804. LUCAS, A. M. AND JAMROZ, C, 1961. ATLAS OF AVIAN HEMATOLOGY. UNITED STATES DEPARTMENT OF AGRICULTURE, WASHINGTON. LUCY, J. A. AND DINGLE, J. T., 1964. FAT-SOLUBLE VITAMINS AND BIOLOGICAL MEMBRANES. NATURE 204:156-160. LUFT, J. H., 1961. IMPROVEMENTS IN EPOXY RESIN EMBEDDING METHODS. J. BIOPHYS. BIOCHIM. CYTOL. 9:409-414. MADSEN, L. L., 1936. THE COMPARATIVE EFFECTS OF COD LIVER OIL, COD THE DEVELOPMENT OF NUTRITIONAL MUSCULAR DYSTROPHY. J. NUTRITION 11:471-494. MALAMED, S., 1963. USE OF A MICROCENTRIFUGE FOR PREPARATION OF ISOLATED MITOCHONDRIA AND CELL SUSPENSIONS FOR ELECTRON MICROSCOPY. J. CELL BIOL. 18:696-700. MALOOF, F. AND SOODAF, M., 1963. INTERMEDIARY METABOLISM OF THYROID TISSUE AND THE ACTION OF DRUGS. PHARMACOL. REV. 15:43-95. MARCH, B. E., BIELY, J. AND COATES, V., 1968. THE INFLUENCE OF DIET ON TOXICITY OF THE ANTIOXIDANT 1,2-DIHYDRO-6-ETHOXY-2,2,4-TRI METHYL QUINOLINE. CANAD. J. PHYSIOL. PHARMACOL. 46:145-149. MASON, K. E., 1949. VITAMIN E. ANN. N. Y. ACAD. SCI. 52:63-428. MATTILL, H. A., 1952. VITAMIN E. NUTRITION REV. 10:225-226. MELLETTE, S. J. AND LEONE, L. A., 1960. INFLUENCE OF AGE,'SEX, STRAIN OF RAT AND FAT SOLUBLE VITAMINS ON HEMORRHAGIC SYNDROMES IN RATS FED IRRADIATED BEEF. FEDERATION PROC. 19:1045-1049. 57 MOORE, T., 1959. VITAMIN E AND UBIQUINONE. NATURE 184:607-608. NADLER, N. AND LEBLOND, C. P., 1955. THE SITE AND RATE OF FORMATION OF THYROID HORMONE. BROOKHAVEN SYMPOSIA 7:40-60. OLIVEIRA, M., WEGLICKI, W. B., NASON, A. AND NAIR, P. P., 1969. DISTRIBUTION OF OC-TOCOPHEROL IN BEEF HEART MITOCHONDRIA. BIOCHIM. BIOPHYS. ACTA 180:98-113. PREMACHANDRA, B. N., PIPES, G. W. AND TURNER, C. W., 1958. THYROXINE SECRETION RATES OF TWO STRAINS OF NEW HAMPSHIRE CHICKENS SELECTED FOR HIGH AND LOW RESPONSE TO THIOURACIL. POULTRY SCI. 37:399-404. PUDELK I EW I CZ, W.' J., WEBSTER, L. AND MATTERSON, L. D., 1964. EFFECTS HIGH LEVELS OF DIETARY VITAMIN A ACETATE ON TISSUE TOCOPHEROL AND SOME RELATED ANALYTICAL OBSERVATIONS. J. NUTRITION 84:113-1.17. QUICK, A. J., 1936. ON VARIOUS PROPERTIES OF THROMBOPLASTIN. AMER. J. PHYSIOL. 114:282-296. QUICK, A. J., 1961. CLINICAL INTERPRETATION.OF THE ONE-STAGE PROTHROMBIN TIME. CIRCULATION 24:1422-1428. REYNOLDS, E. S., 1963. THE'USE OF LEAD CITRATE AT HIGH PH AS AN ELECTRON APAGUE STAIN IN ELECTRON MICROSCOPY. J. CELL BlOL. 17:208-212. RODERUCK, C. E., BASIUSKI, D. H. AND BARBAR, N. A., 1949. SOME CHEMICAL AND ENZYMIC ALTERATIONS IN MUSCLES IN EXPERIMENTAL DYSTROPHY. ANN. N. Y. ACAD. SCI. 52:156-162. ROELS, 0. A., TROUT, M. AND GUHA, A., 1965. THE EFFECT OF VITAMIN A DEFICIENCY AND DIETARY C(r T 0C OPHE R OL ON THE STABILITY OF RAT-LIVER LYSOSOMES. BIOCHEM. J. 97:353-359. ROELS, 0. A., 1967. PRESENT KNOWLEDGE OF VITAMIN E. NUTRITION REV. 25:33-37. ROSE, C. S. AND GYORGY, P., 1950. TOCOPHEROL REQUIREMENTS OF RATS BY MEANS OF THE HEMOLYSIS TEST. PROC. SOC. EXP. BIOL. MED. 74:411-415. ROSE, C. S. AND GYORGY, P., 1952. SPECIFICITY OF HEMOLYTIC REACTION IN VITAMIN E DEFICIENT ERYTHROCYTES. AMER. J. PHYSIOL. 168:414-420. ROSENBERG, L. L., LAROCHE, G. AND DIMICK, M. K., 1963. THYROID FUNCTION IN CHICKS AND RATS. EQUILIBRATION OF INJECTED IODIDE WITH EXISTING THYROIDAL IODINE IN WHITE LEGHORN COCKERELS. ENDOCRINOLOGY 72:759-763. 58 ROSENBERG, L. L., GOLDMAN, M., LAROCHE, G. AND DIMECK, M. K., 1964. THYROID FUNCTION IN RATS AND CHICKS. EQUILIBRATION OF INJECTED IODIDE WITH EXISTING THYROIDAL IODINE IN LONG-EVANS RATS AND WHIT.E LEGHORN CHICKENS. ENDOCRINOLOGY 74:212-225. ROSENBERG, L. L., LAROCHE, G. AND EHLERT, J. N., 1966. EVIDENCE FOR HETEROGENIous TURNOVER OF IODINE IN RAT THYROID GLANDS. ENDOCRINOLOGY 79:927-934. ROSENBERG, B. AND JENDRASIAK, G. L., 1968. SEMICONDUCTIVE PROPERTIES OF LIPIDS AND THEIR POSSIBLE RELATIONSHIP TO LIPID BILAYER CONDUCTIVITY. CHEM. PHYSIC LIPIDS 2:47-54. ROSENKRANTZ, H., 1955. STUDIES IN VITAMIN E DEFICIENCY. 1. THE OXYGEN CONSUMPTION OF VARIOUS TISSUES FROM THE RABBIT. J. BIOL. CHEM. 214:789-797. SCHWARZ, K., 1960. FACTOR 3, SELENIUM AND VITAMIN E. NUTRITION REV. 18:193-197. SCOTT, M. L., NESHEIM, M. C. AND YOUNG, R. J., 1969. IN "NUTRITION OF THE CHICKEN" PP. 144-160. PUBLISHED BY M. L. SCOTT AND ASSOCIATES, ITHACA, NEW YORK. SILVER, S., YOHALEM, S. B. AND NEWBURGER, R. A., 1955. PITFALLS IN DIAGNOSTIC USE OF RADIOACTIVE IODINE. J. AMER. MED. ASSOC. 159:1-5. STAHL, P., TURNER, G. W., 1961. SEASONAL VARIATION IN THYROXINE SECRETION RATE IN TWO STRAINS OF NEW HAMPSHIRE CHICKENS. POULTRY SCI. 40:239-242. STASILLI, N. R., KROC, R. L. AND EDLIN, R., 1960. SELECTIVE INHIBITION OF THE CALORIGENIC ACTIVITIES OF CERTAIN THYROXINE ANALOGUES WITH CHRONIC THIOURACIL TREATMENT IN RATS. ENDOCRINOLOGY 66:872-885. STEMPAK, J. G. AND WARD, R. T., 1964. AN IMPROVED STAINING METHOD FOR ELECTRON MICROSCOPY. J. CELL BIOL. 22:697-701. SURE, B., 1924. DIETARY REQUIREMENTS FOR REPRODUCTION. J. BIOL. CHEM. 58:693-709. SURE, B., 1924. DIETARY REQUIREMENTS FOR REPRODUCTION. J. BIOL. CHEM. 62:371-396. SURE, B., 1925. DIETARY REQUIREMENTS FOR REPRODUCTION. 63:211-223. J. BIOL. CHEM. 59 SWANSON, M. A. AND ARTOM, C, 1950. THE LIPID COMPOSITION OF THE LARGE . GRANULES (MITOCHONDRIA) FROM RAT' LIVER. J. BIOL. CHEM.' 187:281-286. TAPPEL, A. L. AND ZALKIN, H., 1959A. INHIBITION OF LIPID PEROXIDATION IN MITOCHONDRIA BY VITAMIN E. ARCH. BIOCHEM. BIOPHYS. 80:333-336. TAPPEL, A. L. AND ZALKIN, H., 1959B. LIPID PEROXIDATION IN.ISOLATED MITOCHONDRIA. ARCH. BIOCHEM. BIOPHYS. 80:326-332. TAPPEL, A. L. AND ZALKIN, H., 1960. INHIBITION OF LIPID PEROXIDATION IN MICROSOMES BY VITAMIN E. NATURE 185:35-36. TAPPEL, A. L., 1962. VITAMIN E AS THE BIOLOGICAL LIPID ANTIOXIDANT. VITAMINS AND HORMONES 20:493-510. TATA, J. R., ERNSTER, L., LINDBERG, 0., ARRHENIUS, E., PEDERSEN, S. AND HEDMAN, R., 1963. THE ACTION OF THYROID HORMONES AT THE CELL LEVEL. BIOCHEM. J. 86:406-428. TAUROG, A., CHAIKOFF, K. AND FELLER, D. D.,. 1947. THE MECHANISM OF IODINE CONCENTRATION BY THE THYROID GLAND: ITS NON-ORGANIC IOD INE-BI ND ING CAPACITY IN THE NORMAL" AND PROPYLTHI 0URAC I L-TRE ATED RAT. J. BIOL. CHEM. 171:189-201. TSEN, C. C. AND COLLIER, H. B., 1960. THE PROTECTIVE ACTION OF TOCOPHEROL AGAINST HEMOLYSIS OF RAT ERYTHROCYTES BY DIALURIC ACID. CANAN. J. BIOCHEM. PHYSIOL. 38:957-964. UMBREIT, W., BURRIS, R. H. AND STAUFFER, J. F., 1959. MANOMETRIC TECHNIQUES AND TISSUE METABOLISM. BURGESS PUBLISHING CO., MINNEAPOLIS. VALENTI, G. AND BOTTARELLI, E., 1965. RADIOISOTIPIC AND CHROMATOGRAPHIC STUDY OF THYROID FUNCTION AFTER EXCESS OF VITAMIN E IN THE RAT. FOLIA ENDOCRINOL. 18:318-326. CITED IN NUTRITION ABSTRACTS AND REVIEWS. 36(3):717 (1966). 131 VANARSDEL, P. P. AND WILLIAM, R. H., 1956. ALTERATION IN THYROXINE-I METABOLISM PRODUCED BY THYROIDECTOMY, HYPOPHYSECTOMY AND ADRENALECTOMY. AMER. J. PHYSIOL. 185:77-80. VANDERLAAN, W. P. AND BESSELL, A. M., 1946. EFFECTS OF PROPYLTHIOURACIL AND POTASSIUM THIOCYANATE ON THE UPTAKE OF IODINE BY THE THYROID GLAND OF THE RAT. ENDOCRINOLOGY 39:157-160. VANDERLAAN, J. E. AND VANDERLAAN, W.P., MECHANISM OR THE RAT THYROID AND ENDOCRINOLOGY 40:403-416. 1947. THE IODIDE CONCENTRATING ITS INHIBITION BY THIOCYANATE. 60 VAN VLEET, J. F., HALL, B. V. AND SIMON, J., 1968. VITAMIN E DEFICIENCY. AMER. J. PATH. 52:1067-1077. VASINGTON, F. D., REICHAR, S. M. AND NASON, A., 1960. BIOCHEMISTRY OF VITAMIN E. VITAMINS AND HORMONES 18:43-87. VICTOR, J., 1934. METABOLIC AND IRRITABILITY CHANGES IN NUTRITIONAL MYOPATHY OF RABBITS AND DUCKS. AMER. J. PHYSIOL. 108:229-236. WAHLBERG, P., 1955. THE EFFECT OF THYROTROPIC HORMONE ON THYROID FUNCTION. A COMPARATIVE STUDY ON CHICKS WITH RADIOACTIVE INDICATORS. ACTA ENDOCRINOL. SUPP. 23:1-52. WILLIAMS, E. D. AND VICKERY, A. L., 1965. STUDIES ON THE EARLY STAGES OF THYROIDAL IODIDE CONCENTRATION AND BINDING IN ADULT RAT THYROID. LAB. INVEST. 14:1939-1945. Wiss, 0., BUNNELL, R. H. AND GLOOR, U., 1962. ABSORPTION AND DISTRIBUTION OF VITAMIN E IN THE TISSUE. VITAMINS AND HORMONES 20:441-455. WITTING, L. A. AND HORWITT, M. K., 1962.. RAT TOCOPHEROL NEEDS AS FUNCTION OF N IN (CH2CH=CH)N. FEDERATION P.OC. 21:474. WYNN, J., 1968A. THYROXINE DEGRADATION ANTIOXIDANT FUNCTION AND NONENZYMATIC DEGRADATION DURING MICROSOMEAL LIPID PEROXIDATION. ARCH. BIOCHEM. BIOPHYS. 126:880-891. WYNN, J., 1968B. DEGRADATION AND FUNCTION OF THYROXINE. CLIN. ENDOCRINOL. 2:165-176. ZALK I N, H. AND TAPPEL, A. V ITAMIN E ACT I ON. DEFICIENT RABBIT. L., 1960. STUDIES OF THE MECHANISM OF IV. LIPIDE PEROXIDATION IN THE VITAMIN E ARCH. BIOCHEM. BIOPHYS. 88:113-117. TABLE 1.--BASAL DIET IN EXPERIMENT IA. INGREDIENTS % GROUND WHEAT 83.0 HERRING MEAL 14.5 DISTILLERS' DRIED SOLUBLES 2.0 IODIZED SALT 0.5 TOTAL 100.0 TABLE 1B. — BASAL DIET IN EXPERIMENT 1B. INGREDIENTS GROUND WHEAT 81 .5 HERRING MEAL 14.BONE MEAL 0.5 LiME STONE 1 .0 DISTILLERS' DRIED SOLUBLES 2.0 IODiZED SALT 0.5 TOTAL 100.0 1_ ' EACH KG. or BASAL DIET CONTAINED THE FOLLOWING INGREDIENTS: MANGANESE SULPHATE, 12.54 MG; VITAMIN A, 5505 I.U.; VITAMIN D^, 440 I.U., MENADIONE, 0.60 MG.; RIBOFLAVIN, 3.61 MG.; CALCIUM PANTOTHENATE, 10.21 MG.; NIACIN, 10.12 MG.; FOLACIN, 1.21 MG.; CHOLINE CHLORIDE, 128.7 MG.J AMP R OL IUM, 0.125 MG. 62 TABLE; 2.--AVERAGE: CH1CKS THYROID WEIGHTS, IN EXPERIMENT 1 A . BODY WE IGHTS AND FEED CONSUMPTION OF TEMPERATURE c° CONTROL EXCESS-V1TAM1N E EQ THYROID WEIGHT MG. 14.5 C° 31.5 C° 28.13±5.53 21.67±4.27 28.70±2.86 22.14+5.40 19.29±5.07 18.50±4.08 BODY WEIGHT GM . 14.5 C° - 31.5 C° 340±21.8 341±30.5 359±16.5 335±25.9 161+12.4 187±16.5 THYROID WT. MG./100GM.B.W. 14.5 C° 31.5 C° 8.25+1.31 6.35±1.09 8.09±0.67 6.55±1.20 11.92±2.80 11.5112.57 FEED CONSUMED PER CHICK IN 13 DAYS GM. c 14.5 C° 31 .5 C° 401 341 454 341 242 168 TEMPERATURE TH1OURAC 1 L EXCESS V1TAM1N E + THIOURAC 1 L EQ +TH1OURACIL THYROID WEIGHT MG . 14.5 C° 31.5 C° 123.26±56.20 95.96±16.49 85.02±52.74 72.43±17.51 123.30±42.50 71.57±23.37 BODY WEIGHT GM . 14.5 C° 31 .5 C° 316±27.7 335±28.8 280±28.5 321±29.2 176±14.9 183±13.8 THYROID WT. MG./DOGM.B.W. 14.5 C° 31.5 C° 38.16±14.27 28.65±6.20 29.37±15.66 22.70±5.20 68.86±16.01 39.53±14.73 FEED CONSUMED PER CHICK IN 13 DAYS GM. 14.5 C° 31.5 C° 378 325 310 250 272 227 63 TABLE 3.--AVERAGE THYROID WEIGHTS, BODY WEIGHTS, FEED CONSUMPTION OF CH i CKS IN EXPERIMENT IB. EXCESS TEMPERATURE CONTROL V 1 TAM1N E THYROID WEIGHT 14.5 C° 25.02±3.46 25.98±6.00 MG. 31 .5 C 23.26±3.16 22.68±4.66 BODY WEIGHT 14.5 C° 364±22 369±38 GM. 31.5 C° 376±41 373±33 THYROID WT. 14.5 C° 6.87±0.84 7.02±1.24 MG./100GM.B.W. 31 .5 C° 6.20±0.64 6.05±0.89 FEED CONSUMED PER • CHICK IN 13 14.5 C° 549 561 DAYS GM. 31.5 C° . 362 357 EXCESS V1TAM1N E TEMPERATURE TH IOURACIL +TH1OURAC 1 L THYROID WEIGHT 14.5 C° 123.10±25.30 105.63±16.44 MG . 31.5 C° 109.31t25.03 94.06±16.69 BODY WEIGHT 14.5 C° 335±35 342±34 GM. 31.5 C° 350+31 353±32 THYROID WT. 14.5 C° 34.03±5.11 30.98±4.07 MG./100GM.B.W. 31 .5 C° 31.09±5.27 26.63±4.36 FEED CONSUMED PER CHICK IN 13 DAYS GM. 14.5 C° 31.5 C° 423 328 416 333 64 TABLE 4.--ANALYSIS or VARIANCE OF THYROIDAL RESPONSE TO THE EXCESS VITAMIN E AND/OR THIOURACIL AT DIFFERENT TEMPERATURES IN EXPERIMENT 1A. SOURCE DF MEAN SQUARE F TEMPERATURE (A) 1 292.842 4.611* VITAMIN E (B) .1 165.169 2.601 THIOURACIL (C) 1 6005.450 94.560** AB 1 2.799 0.044 BC 1 .167.179 2.632 AC 1 124.614 1 .962 ERROR 41 63.509 TOTAL .47 SIGNIFICANT (P «=0.05). HIGHLY SIGNIFICANT (P < 0.01). 65 TABLE 5.--ANAL.YS I S OF VARIANCE OF THYROIDAL RESPONSE TO THE EXCESS VITAMIN E AND/oR THIOURACIL AT DIFFERENT TEMPERATURES IN EXPERIMENT 1B. SOURCE DF MEAN SQUARE F TEMPERATURE (A) 1 103.937 5.982* V1TAMIN E (B) 1 166.519 9.584** THIOURACIL (C) 1 17523.512 1008.606** AB 1 0.164 0.0094 BC 1 •178.202 10.257** AC . 1 238.754 13.742** ERROR 109 17.374 TOTAL 115 SIGNIFICANT (P<0.05). HIGHLY SIGNIFICANT (P <0.01). 66 TABLE 6. — AVERAGE THYROID WEIGHTS, BODY WEIGHTS, FEED CONSUMPTION OF EQ-TREATED CHICKS AT 22 C° IN EXPERIMENT 1B. EQ EQ +TH1OURACIL CONTROL TH1OURAC I L THYROID WEIGHT MG . 25.90±6.09 138.28±34.14 28.73±7.11 49.82±11.83 BODY WEIGHT GM. 307±47 313±36 353±41.4 380±30 THYROID WT. MG./100GM.B.W. 8.50±1.67 43.67±7.7CV 8.09±1.58 13.22±3.45 FEED CONSUMED PER CHICK IN 13 DAYS GM. 292. 302 333 318 67 TABLE 7.--EFFECT OF EXCESS DIETARY VITAMIN E ON THYROIDAL IODINE (l ) UPTAKE AND RELEASE IN WHITE LEGHORN MALE CHICKS. A. MEAN VALUES, THYROIDAL LEVEL OF J31 of 1 PER BIRD:$ OF INJECTED DOSEiS.D 5 HR 10 HR 24 HR 48 HR 96 HR BASAL 11.75±1.52 10.68±2. 03 11.17±3.25 9.60±2.57 7.17±2. 19 EXCESS V1TAMIN E 8.11±1.69 9.35±2. 19 10.26±2.24 9.14±2.16 9.86±2. 68 B. MEAN VALUES, THYROIDAL LEVEL OF I 131 I ifo OF INJECTED DOSE/MG. TH YRO I D±S .D. 5 HR 10 HR 24 HR 48 HR 96 HR BASAL 0.427±0.061 0.433±0.037 EXCESS VITAMIN E 0.306±0.047 0.363±0.081 0.466±0.063 0.355±0.059 0.317±0.045 0.362±0.04b 0.323±0.053 0.342±0.057 68 TABLE 8.--RESPIRATION RATES OF MUSCLE MITOCHONDRIA FROM THE CHICKS FED THE CONTROL DIET, EXCESS VITAMIN E AND/OR THIOURACIL. INCUBATION WAS AT 30°C. I NCUBATI ON PERiOD CONTROL DIETARY TREATMENT EXCESS V t TAMiN E THIOURACIL EXCESS VITAMiN E +THIOURACIL 0-30 MIN. 30-60 MIN. 60-90 MIN. 90-120 MIN. TOTAL UL/HR/MG. PROTEIN (7)° 11.80±2.78 12.03±1 .18 10.71 ±3.19 7.01±2.62 (?) 7.05±3.28 7.70±3.36 8.03±4.65 4.64±2.20 41.55±8.22A 27.42±12.81B 20.78±4.11A 13.71±6.41 B (6) ' 8.50±3.85 8.84±2.89 8.51±2.62 6.35±2.47 32.20±11.67' 16.10±5.84A (7) 5.37±1.94 4.84±2.21 4.44±1.91 3.97±2.06 18.63±7.64 9.32±3.82E B BODY WEIGHT (GM. ) 597±91 363±60 482±66 332±40 EXPRESSED AS UL 0^ UPTAKE PER MG. MITOCHONDRIAL PROTEIN±S.D. DURING SUCCESS.! VE 30 MINUTE PERIODS. "VALUES WITHIN A LINE HAVING THE SAME SUPERSCRIPT ARE NOT STATISTICALLY DIFFERENT AT THE 5$ LEVEL OF PROBABILITY BY THE DUNCAN'S MULTIPLE RANGE TEST. ^NUMBER OF CHICKS TESTED. TABLE 9.--COMPOSITION OF THE EXPERIMENTAL DIETS. DIET 1 BASAL' DIET 2 +C A DIET 3 +EXCESS +VITAMIN E D I ET 4 +EXCESS +V I TAMIN E +CA % % % BASAL DIET 98.0 98.0 98.0 98.0 © CELLULOSE 2.0 2.0 BONE MEAL 1.5 1 .5 LIME STONE 0.5 0.5 TOTAL 100.0 100.0 100.0 100.0 DL-OC-TOCOPHERYL ACETATE (|.U./KG) 10. 10. 2200 2200 BASAL DIET WAS THE SAME AS SHOWN IN TABLE 1A. TABLE 10...— E-FFECT OF EXCESS VITAMIN E ON HEMATOCRIT VALUE, PROTHROMBIN TIME AND RECT I CULOC YTE COUNT. AGE OF CHICKS (DAYS ) BASAL BASAL +CALCiUM DIET EXCESS V ITAMIN E EXCESS VITAMiN E +CALCIUM HEMATOCRIT ± S.D. 30 40 50 24.94±1.57 25.08±1.50 27.63±1.78 28.06±0.81 B 27.69±0.72 27.00±2.18 14.51+2.57 13.81 ±3.54 B 21 .67+2.18 B 18.13±0.78 15.89±2.37 23.58±2.06 B B PROTHROMBIN TIME (SECONDS) 30 40 56.94±4.90 43.44±5.19 51.90±6.72 38.28±5.58 96.36±12.44 B 141.60±18.78 92.34±9.75 B 105.00±18.33 RECTICULOCYTE ± S.D. 30 40 50 1.73±0.95 12.98±2.70 11.92±3.25 10.68±1 .02 9.65±0.73 11.64+1.20 47.45±3.99 B 42.08±9.37 B 52.44±5.02 44.50±9.17 B 40.60±19.07 24.38±3.83B B 1 VALUES WITHIN A LINE HAVING THE SAME SUPERSCRIPT ARE NOT STATISTICALLY DIFFERENT AT THE 1$ LEVEL OF PROBABILITY BY THE DUNCAN'S NEW MULTIPLE RANGE TEST. 71 TABLE 11 --PROTHROMBIN TIME OF CHICKS FED EXCESS VITAMIN E, 24 HOURS AFTER INJECTION OF MENADIONE. EXCESS YITAMiN E EXCESS (24 HR AFTER INJ. CONTROL VITAMIN E OF MENADIONE) PROTHROMBIN TIME (SECONDS) 34.7±3.8 77.9±15.9 27.1±3.2 72 TABLE 12. — EFFECT OF EXCESS VITAMIN E ON BONE CALCIFICATION IN CHICKS FED DIETS ADEQUATE AND DEFICIENT IN CALCIUM. AGE OF CHICKS BODY (DAYS) WEIGHT ASH$ 30 229±81.7 35.74±4.04 GROUP 1 ' BASAL 40 333±75 42.14±0.52 50 497+110 43.97±3.22 30 169±43 31 .6113.43 BASAL GROUP 2 EXCESS 40 202±37 36.76±2.42 +V1TAM1N E 50 338.9±38 37.83±2.13 30 257±23 45.94±0.87 BASAL GROUP 3 + CALC1UM 40 307+42 46.71±0.90 50 513±48 47.20±2.39 30 231±28 45.54±0.56 BASAL GROUP 4 +EXCESS 40 250±32 48.20±1.36 V1TAM1N E +CALC1UM 50 416±55 46.92±0.66 

Cite

Citation Scheme:

        

Citations by CSL (citeproc-js)

Usage Statistics

Share

Embed

Customize your widget with the following options, then copy and paste the code below into the HTML of your page to embed this item in your website.
                        
                            <div id="ubcOpenCollectionsWidgetDisplay">
                            <script id="ubcOpenCollectionsWidget"
                            src="{[{embed.src}]}"
                            data-item="{[{embed.item}]}"
                            data-collection="{[{embed.collection}]}"
                            data-metadata="{[{embed.showMetadata}]}"
                            data-width="{[{embed.width}]}"
                            async >
                            </script>
                            </div>
                        
                    
IIIF logo Our image viewer uses the IIIF 2.0 standard. To load this item in other compatible viewers, use this url:
http://iiif.library.ubc.ca/presentation/dsp.831.1-0102205/manifest

Comment

Related Items