102 717 THS THE EFFECT OF DIET ON- URINARY SULFUR EXCRETION . OF PRESCHOOL CHILDREN Thesis for the Degree of M. 5. MICHIGAN STATE COLLEGE Gladys P. Bond 1940 aid 33 M32 E> “7 071 r'\ "E’ .H.‘ ‘ ”LS Wewu 3 \HA TL . \ . ‘ , NV :" THE EEEECT 0? DIET 0N URINARY SULFUR EXCRETION OF PRESCHOOL CHILDREN by Gums PAULINE gem) A.THESIS Submitted to the Graduate School of Michigan State College of Agriculture and.Applied Science in partial fulfilment of the requirements for the degree of MASTER OF SCIENCE Department of reeds and Nutrition Division of Home Iconomice 191m THEEAS 11 ACKNOWLEDGMENT The writer wishes to express her sincere appreciation to several persons who have contributed greatly to this problem: to Dr. Jean Hawks for her helpful supervision of the study, to Dr. Marie Dye for her suggestions and criticisms. and to Barbara Veley who compiled the nitrogen elimination figures “Bade {Ila IQ «2 £3 3 111 TABLE OF CONTENTS Chapter Page ACKNOWLEDGMENT ......... ................. 11 TABLE OF CONTENTS .. ...... ...................... iii LISTOFTABLES. .. .......... ...... iv LIST OF CHARTS ................................. vi I INTRODUCTION ................................... 1 II RIVIEI or LITERATURE ....... ..... ............... 3 III EXPERIHEITAL PROCEDURE ......................... 7 IV PRESENTATION OF DITA,AND DISCUSSION ............ l5 Urinary Sulphur Excretion 01138881 m.t eeeeeeeeeeeeeeeeoeeeeeeoe. 15 Urinary Sulphur Excretion on High Fat Diet eeeeeeeeeeeeoeeeeeeeeee 8 Urinary Sulphur Excretion on High Staten met Oeeoeeeeeeeeeeee'eee ‘4']. Urinary Sulphur Excretion on High Sugar Diet ..................... 53 v SW .000... eeeeeeee 000000000000.000.000.090. 6n BIEIOGmm Q0000000000....0.000000000000000so. 65 Number \Jl-F-‘WN 10 11 12 13 11+ 15 LIST OF TABLES COMPARISON OF PHYSICAL MEASUREMENTS OE CHEM 'ITH BAIJDWINOVOOD STMDABDS e e e e e e e e e e e 0 DAILY ROUTINE OF CHILDREN . . DAILY BASAL DIET ........... .00... ...... 000......00 OOOOOOOOOOOOOOOOOOOOCCO DIET FACTORS AS BASED ON INDIVIDUAL WEIGHTS ....... PREL IMINARI TESTS TO DETERMINE PERCENTAGE “cm or smmgBa Son .OOOOQOOOOOOOOOOOOO. PERIOD VARIATION OF TOTAL URINARY SULFUR ON BASH! DIET .OOOOOOOOOCCCOOOOOCOOOOOOOOOOOOOOOOOOO SULFUR INTAKE AND URINARY EXCRETION PER KILO— GRAM OJ' BODY WEIGHT ON BASAL DIET ............... VARIATION OF URINARY SULFUR EXCRETION IRON PERIOD GROUP AVERAGE ON BASAL DIET ....... ....... CHANGES IN WEIGHT AND PERCENTAGE OF SILEUR mum ON BASH) DIM .00OOOOOOOOOOIOOOOOOOOOOOO N/s MIG ON 35% D1“ .0..OOOOOOOIOOOOOOOIOOOOOOO PERIOD VARIATION OF TOTAL URINARY SULFUR ON HIM rm DIM .0000....OOOOOOOOOOOOOOOO0.0.0.0... CHANGES IN IEIGHT AND PERCENTAGE OF SULFUR EXCRETED ON HIGH EAT DIET Oeoeeeeeooeeeeeeeeeeeeo SUIJUR INTAKE AND URINARY EXCRETION PER KILO- GRAM OF BODY WEIGHT 0N HIGH FAT DIET ............ VARIATION OF URINARY SULFUR EXCRETION IRON PETIOD GROUP AVERAGE 0N HIGH FAT DIET ........... N/S mm on HIGH rm our iv Page 8 9 10 11 1h 16 19 2’4 25 27 30 33 3h 38 16 17 18 19 8 22 23 2h 25 26 2? PERIOD VARIATION OF TOTAL URINARY SULFUR ON HIGH STARCH DI” 00.00....0..0...0000 SULFUR INTAKE AND URINARY EXCRETION PER KILO- GRAN 01' BODY [EIGHT ON HIGH STARCH DIET ..... .... VARIATION OF URINARY SULFUR EXCRETION FROM PERIOD GROUP AVERAGE ON HIGH STARCH DIET .. ...... CHANGES IN NEIGHT AND PERCENTAGE Ol' SULFUR EXCRETED ON HIGH smaca DIET N/S mm ON HIGH STARCR pm RANGE IN VARIATION or H/s RATIOS ON HIGH STECH DIET 00000000000000000000 0.0.0.000... PERIOD VARIATION OF TOTAL URINARY SULFUR ON HIGH 3mm DIET .00....00.0.0000000.. 0.0.0.000... SULJ'UR INTAKE AND URINARY EXCRETION PER KILO. GRAN OF BODY WEIGHT ON HIGH SUGAR DIET| VARIATION OF URINARY SULFUR FROM PERIOD GROUP AVERAGE ON HIGH SUGAR DIET .. .. CHANGES IN WEIGHT AND PERCENTACE OF SULFUR EXCRETED ON HIGH SUGAR DIET N/S mm ON HIGH SUGAR DIET RANGE IN N/S RATIOS ON HIGH SUGAR DIET ...0000.0.0. 1‘3 1*5 “9 n9 52 52 5h 58 61 62 62 Chart Number 10 11 12 13 1h 15 LIST OF CHARTS Page Total Sulfur Excretion on Basal Diet ............ 17 Sulfur Excretion Per Kilogram of Body 'ei@tonBasa]-D1°t .00.000.000.00000000000000 20 Variations of Sulfur Dxcretions From Individual Averages On Base]. Diet . . . . . . . . . . . . . 22 Individual Variation in Excretion Iron Period Group Average On Basal Diet . . . . . . . . . . . . 23 N/S Ratio On Basal Diet ......................... 28 Total Sulfur Excretion On High Fat Diet ......... 32 Sulfur Excretion Per Kilogram of Body ‘eight on High Fat Diet 00......00000000000000. 35 Variations ofISulfur Emretione Iron Individual Averages On High Fat Diet . . . . . . . . . . 37 Individual Variation in Excretion Prom Period Group Average On High Fat Diet .. ....... 39 N/s htio on Big- rat Diet .0...0..........0.00.. 39 Total Sulfur Excretion On High Starch Diet ...... he Sulfur Excretion Per Kilogram of Body weight onnigh st”ChD1°t .000...0.0.0.00.00.. ”‘6 Variations of Sulfur Excretion: From Individual Averages On High Starch Diet ....... 1L8 Individual Variation In Excretion From Period Group Average On High Starch Diet .......h8 N/S Ratio On High Starch Diet ................... 51 vi 16 17 18 19 vii Total Sulfur Excretione On High SW Diet 0000.00.0000000000000000000.0..0000. 55 Sulfur Excretion Per Kilogram of Body Weight On High Sugar Diet ..................... 57 Variations of Sulfur Excretions From Individual Averages On High Sugar Diet ........ 59 Individual Variation in Excretion From Period Group Arerage On High Sugar Diet ....... 59 Ms Ratio On High Sugar Diet 63 INTRODUCTION This report on urinary sulfur excretion is part of a study done in an effort to add to our present knowledge of the mineral metabolism of preschool children. While sulfur is one of the inorganic ions essential to normal nutrition. few studies have been made to determine its functions. This lack of experimentation may be due to several factors. First, sulfur is found in the body in the form of complex organic compounds rather than an inorganic substance. The majority of these compounds are protein in nature and complicated in structure. Thus the determination of sulfur is not a simple procedure. Since sulfur is in combination with the protein and the amounts are quite nearly proportionate, it has been assumed.that the body obtained suf- ficient sulfur if the diet contained adequate protein. Nevertheless, sulfur is a small but integral part of the living organism, as shown by Dr. H. B. Levis' chart:(1) Sulfur Compounds in the living organism A. Naturally occurring organic sulfur compounds containing nitrogen 1. Proteins (0.2 to 5.0 per cent 8) 2. Known constituents of the protein.lolecule - The sulfur- containing amino aside a. sttine - lollaston - lSlO-Calculi - Iorner 1899-proteiu b. Methionine - Mueller - 1923 - Berger and Coyne - 1928 3. Probable derivatives of protein or of the above amino acids a. Taurine (taurochloric acid) b. Cysteine c. Ergothioneine a. Glutathione (reduced and oxidized) .e Insulin f. Hercapturic acids 3. Compounds related to or derivatives of naturally occurring sulfur compounds (the biological significance of these remains to be proven) l. Homologues of cystine or cysteine a. Homocystine (homocysteine) b. Pentocystine (psntoqysteine) 2. Homologues of methionine a. Homomethionine . Thiolhistidine . Products of partial oxidation of qystine and cysteine a. ayataic ”1d b. Suboxidation products of cystine and cysteine. (Toennies - Hammett) Because it forms such an important part of all living tissue, the functions and requirements of sulfur should be studied in detail. The urinary figures presented in the following pages represent one possibility for study-that of total sulfur elimination. Actually. these figures include fractions which could.be determined separately. The larger amount of sulfur is eliminated in the inorganic sulfate sulfur fraction. The ethereal or conjugated sulfuric acids include a.smaller portion of the oxidised.sulfur while the sulfur compounds of organic nature constitute the remainder of the sulfur eliminated. REVIEW 01' LITERATURE Since an adequate protein intake apparently insures an adequate sulfur intake. there have been few studies on sulfur metabolism. In fact. the literature up to the present time contains but one referb ence to a sulfur balance study done on preschool children. so there have been no recommendations as to dietary sulfur requirements. There have been several studies on infants as well as a few scattered re- ports on adults. Many of the adult studies have been made on hospital patients which makes it impossible for the results to be used in com- parison with the normal subjects of this study. Many investigators have reported sulfur balance studies on animals. .A full review of these latter studies will not be included in this paper. however men- tion shall be made only of those which.might have some pertinence to this particular study. ' All investigators seem agreed that the major portion of the sulfur is eliminated in the oxidized form through the kidneys. Shohl and Sato (2 8‘ 3) studied two babies. 7 and 9 months old. and reported that 90$ of the sulfur was eliminated through the kidneys. Blazeo(l938)(u) found from a study of 7 infants aged 1-6 months, and 9 children 3-lh years. that in both groups the urinary sulfur exceeded the fecal exp cretion. Cooley, Penberthy, Armstrong. Hunscher. Macy. and Cape(5) found the average percentage urinary sulfur excretions of two girls (11 and.7% years old) to be 76.h3$ and 76.26% respectively. These results were considerably lower than those presented by Shohl and Beta“2 & 3) for infants. but may merely indicate a higher fecal sulfur excretion by the older children. Some comparative studies of the utilisation of human and cow milk have been made on infants to determine if there are differences in sulfur metabolism. Blends) reported that the infants on his study retained approximately 90% of the sulfur from human milk, 75% from a diet of cow and human milk, and only 60% from a diet of cow milk alone. He also noted that boiling human milk reduced the sulfur re- tention 345%.”) It might be expected that the urinary and fecal sulfur excretions would vary between infants and children. Nevertheless, Blazes)” found that on the basis of body weight the fecal sulfur was of the same con- centration in both groups. On the other hand, he found that the con- centration of urinary sulfur was from four to six times as great in the children as in the infants. Although all human studies are not directly comparable with this report. certain conditions have an effect on the sulfurmstabolism. Goons, Goons. and Schiefelbusch“) found a negative sulfur balance (hiring pregnancy. Aud.conversely, Hunell. Sternberger. Hunscher. and Macy”) reported a mean daily sulfur balance of 0.3h '3 0.13 gm. during the last 116 days of pregnancy. Himmell, Hunscher. Bates, Bon- ner, and “soy found a mean daily storage of 0.27 gms. of sulfur in another primapora observed during the last 65 days of gestation. Since sulfur is so widely associated with protein. the relation- ship is commonly expressed as a I]! ratio. This ratio varied under different conditions of diet. rolin (1905)(11) found that the ratio for an adult man was practically the same on a high and.low protein diet (11.5 and 11.8 respectively). In animal studies Terraine and Rasafimahery (1935)(12) feund that there was a sharp increase in the 11/8 ratio of the excreta of animals going from simple privation of proteins to complete starvation. As starvation continued. there was a slow diminution in the ratio and Just before death a further decline although the total amounts of nitrogen and sulfur were much increased. They interpreted it as meaning that. in the last phase, muscle tissue and sulfur containing amino acids were undergoing dis- integration. The quality of protein apparently had no influence on the 11/8 ratio in hogs since Rajsman (1936)(13) reported the ratio re- mained the same even when the diet contained poor or mediocre quality protein. This ratio may not be retained under all conditions. however. The sulfur excretion is also affected by tissue injuries. Opera- tions, and certain diseases. Outhbertson (1931)“‘0 reported an early catabolic loss of nitrogen and sulfur resulting from tissue injury... the increase in sulfur being due to a sligitly greater proportionate increase in excretion of inorganic sulfate. The N/ S ratio may also (15) be altered in diseased conditions. Grabfield and Prescott noted that in congestive heart failure. orthostatic albuminuria. toxemia of pregnancy and glomerulonephritis, the ratio is approximately 20:1. In the nephrosis syndrome. due to low sulfur. the ratio may go as high as 3081. The determining factor in the sulfur elimination of normal indivi- duals apparently is the protein intake. Although Iolin (11) reported the 1/8 ratio to be the same on a high or low'protein diet, he found that the nitrogen and sulfur excretions were less on the low'protein diet. The excreted sulfur fractions. however. did not vary in the same preportion from a high to low protein diet. The inorganic sul- fate sulfur and conjugated sulfur were reduced on the low protein diet while the organic sulfur excretion was practically the same on both diets. In rabbits Razafimahery (1935)(16) found.that the excre- tion of combined sulfur was slightly increased.when a regime rich in protein replaced a.non-protein one. Blassgh) found that while the infants in his study retained.only “0% of the protein nitrogen absor- bed, they rstained.up to 90% of the protein sulfur absorbed. He con- cluded, therefore, that it is predominantly the sulfur containing con- stituents of the protein which are utilized by the growing organism. Although these studies show general trends in sulfur metabolism of the living organism, human and animal. the data presented is too scattered and incomplete to be conclusive. There is a definite need for more detailed studies of sulfur metabolism. EXPERIMENTAL PROCEDURE This urinary sulfur study was one of a series of studies made to determine whether the addition of calories in different forms had any effect on the protein or mineral metabolism of a group of pre- school children. The six subJects. three boys and three girls. ranged in age from forty-two to fifty-seven months at the beginning of the study. All were in good.physica1 condition as determined by complete medical examinations and by a thorough study of their past medical histories. The nutritional backgrounds of the children were similar inasmuch as four of them (I. I, C. and NJ) had been in the same state institution! since birth. and the other two, J and K. had been together in a boarding home. All had.had some of the usual childhood diseases. but in no instance had there been any irregularity which would indi- cate that any of the normal metabolic processes had been disturbed or impaired. Their height-weight records, as shown in Table l were within the normal range for children of the same height and.age ac- cording to the standards set by Wood and Baldwin.(17) The children lived for the entire three months' experimental peri- od in one of the college houses located on the campus. Since any bal- ance study necessitates the most carefully controlled conditions pos- sible, they followed a definite daily routine in their activities. as presented in Table 2. They were together at all times. being segregated 'Lapeer Home and Training School, Lapser, Michigan. COMPARISON OF PHYSICAL MEASUREMENTS OF CHILDREN WITH TABLE 1 BALDWIN-HOOD STANDARDS ___‘_ :— Baldwin-Food BaldwinAIood ‘Child -Age Height Ayerage Variation Weight Average Variation Height Ieight Months cm. cm. 95 52:... _.!s- l 0 R7 103.3 102.51 +0.86 1N.9u 17.01 -1.22 50 105.5 10N.33 .1.12 17.23 17.h2 -1.09 r 50 105.5 10u.33 .1.12 17.38 17.h2 -0.23 53 107.0 105.98 o0.96 20.13 17.90 e1.25 “I he 102.6 98.88 :3.76 16.19 16.06 +0.81 1‘3 10301 98e88 e 27 16e51 16e06 02e80 J 55 109.6 105.31 oh.o7 16.87 17.18 -0.18 NJ 51 103.h 103.10 +0.29 15.86 16.51 -o.39 5h 105.0 105.31 -o.29 17.63 17.18 e0.26 r 57 101.1 107.23 :a.72 17.17 17.61 .2.50 60 103.6 109.02 .97 21.03 17.95 +1.72 TABLE 2 DAILY ROUTINE Ol' CHILDREN A. l. P. Ii. Hour Activity Hour Activity 6:30-7:00 Awakened. went to 1:00-3:30 Nap toilet. weighed and measured 3:30.h:00 Toilet: dressed H to play 7:00-7:30 Dressed Moo-5:00 Play 7:30-8:15 Breakfast 5:00-5:30 Toilet: bath: 8:l5—9:00 Toilet; prepared to undressed go outdoors or other play acti- 5:30-6:15 Supper vity 6:15-6:16 Play (Quiet) 6:15-7:00 Toilet 9:00-11:00 Play 11:00-11:16 Toilet: prepared for lunch 7800 Bed 11:15-12:30 Lunch 12330-1800 Toilet; undressed for map only on two occasions. when a child had a slight cold. The same trained persons supervised their activities constantly. with the aim in view. however. of making this supervision as nonapparent as possible to the children. By employing the same supervision at all times. the inves- tigators hoped to eliminate any possibility of emotional stimuli due to the presence of persons unacquainted with the children and general technique of the experiment. The children seemed happy and well adjust- ed throughout the study. with only an occasional emotion outbreak. which 10 is the natural and normal reaction in a group of children. The children ate four different diets. the only variable being additional calories in different forms. In order that the children might become adJusted to their new food and environment. there was a in day preliminary period. During this time they ate the same foods which were included in the first diet. The first. or basal. diet continued for ten 3-day periods (num- bers l-lO) and contained the foods listed in Table 3. This diet was man: 3 mm BASAL mm:- l‘ood 'eight (an) Hill: 800 Ralstons 20 Orange Juice 200 Beef no Peaches 150 Applesauce 150 Celery' 20 Beans 100 Tomatoes 100 Potatoes 80 Butter 20 sugar 20 Bread 60 Eggs to Water 800 ‘Diet fed to heaviest child.l on basis of 95 cal. 1 kg. body wt. 3 g. prot. 1 kg. bow wt. 11 calculated.for the heaviest child, E. on the basis of 95 calories and 3 grams of protein per kilogram body weight. The other children received diets calculated as fractions of that fed to subject I. (Table h). Thus each child received the same amount of food in referb ence to his weight; for example. each child drank the same amount of milk per kilogram. And since the childrens' weights varied. their diet factors were recalculated at the beginning of each new diet period. TABLE h DIET Empress AS BASED ON INDIVIDUIL wzxcnms* Child. Basal High Fat High Starch High Sugar 0 .8h .82 .82 .89 s 1.0 .95 .95 1.0h z .91 - - - J .9“ .9h - - NJ .89 .87 - - r .95 .95 .95 1.06 *The diet factors were determined by the individual weights and.were computed as fractions of the diet (1.0) fed to the heaviest child. I. The second diet period continued for eight 3-day periods (numbers 11-18). During this time filtered butter fat was added to the basal diet daily. thereby introducing a calculated 20% increase in the cal- oric content of the diet. The butter was easily added to the vegeta- bles and cereals. apparently escaping notice on the part of the children. The third diet lasted for the next eight 3-day periods (numbers 19-26). The calculated 20% increase in caloric content over that of 12 the basal diet was effected by the addition of cornstarch. Unlike the my: fat diet. the high starch diet presented some difficulties as to a.method of feeding cornstarch in a.palatable form. Finally. since the children liked cake. the problem was solved by making the corn- starch into individual cakes. While these were not exactly like real cake. the children ate them with no serious objections. The fourth and last diet also continued over eight 3-day periods (numbers 27-3“). The calculated 20% increase in calories was made hy adding sugar to the diet. This modification involved no difficul- ties whatsoever since the sugar was made into candy. The children were very pleased with the change in their diet and seemed more in- pressed by this than any of the other modifications. Every precaution was taken to weigh and feed the diets quantitap tively. The food was weighed in advance for three day periods from the same source of supply. Thus the three day food samples were iden- tical in composition. Duplicate samples of the smallest child's daily intake (0) were saved and dried to constant weight for analysis. The children ate their diets quantitatively. which.included wiping the dishes with.sma11 pieces of bread and rinsing them with distilled water. These washings were then added to the food for the same day. The distilled water intake was also measured for each child.‘ Three of the children made no objection to any of their food throughout the entire experiment. One child. I. was taken off directly _— _ 'The individual daily distilled water intake was as follows: I - 800 gms. I - 723 gms. Y - 760 ' NJ - 707 ' J-m " 6-697 ' 13 following the basal diet because of his complete refusal to cooperate. And while the other two. J and MJ. remained on the study until the end. their excretion figures were not included on the high starch and high sugar diets because they did not eat all of their food in some instances. The difficulties incurred with these girls probably had a.psychological rather an a.physiological basis. The procedure followed for the urinary sulfur analysis was the s<18) modification of the Benedict methodslg) 50 m1! of the three Deni day pooled sample of urine were measured into an evaporating dish and 5 ml?.of the Benedict-Denis oxidizing agent added to it. Triplicate determinations were made on each sample. The samples were placed on an electric plate at low temperature and.evap0rated. Then the temperap ture was turned up through medium to high.heat and left until the mix, ture was thoroughly charred. To insure complete oxidation. the evapo- rating dish and contents were placed in the muffle furnace at 750° for 20 minutes. removed. cooled. and 20 m1. of 10% £01 added to dissolve the oxidized mixture. This acid solution was then transferred quanti- tatively into a 250 ml. beaker with 100 m1. of distilled water. The contents of the beaker were heated to Just below the boiling point. and 25 ml. of lOfitBaOl added slowly with constant stirring. This was 2 then left on the steam bath for 18 hours or more. filtered through a *The Benedict-Denis method suggests 20 ml. of urine. but due to the dilution of some of the childrent' urine. 50 ml. of urine was used in this study. (In some instances. smaller amounts of urine were used due to nonavailability of larger samples.) *‘Preliminary tests were run. using 5 and 10 ml. portions of the oxi- dising reagent. Since there was no appreciable difference between re- sults obtained from the varying amounts. the 5 ml. portion was used so that there would be less error in the form of additional Na.ions intro- duced. 1h porous bottom crucible and the precipitate washed with hot distilled water. The crucible was next heated in the muffle furnace at 7500 for 20 minutes. cooled. and weighed. The total sulfur figures were cal- culated from the weight of the BaSOM thus obtained. Preliminary tests were run. using Na280n and.la230h plus urine. The average percentage recovery was 99.89%. and in no instance was there more than a 2.6% variation from the theoretical yield (Table 5). arms 5 PRELIMINARY TESTS TO DETERMINE PERCENTAGE RECOVERY OF SULFUR.AS Bason Theoretical Nazsou Urine Urine % Yield 3 Recovery Naesoh .1002 .0985 .. .— 98.2 .1002 .0978 .- - 97.6 .1002 ~ . 2161 .1185 97.1: .1002 -- . 2183 -- 99.6 .1002 .0985 .. .— 98.2 .1002 .0991 v“ "" 9809 .1002 - .2079 .1056 102.1 .1002 .- .2082 -. 102.1: .1125 .1132 ~ - 100.6 .1125 .11h2 - -. 101.5 .1125 - .2307 .1158 102.1 .1125 .- .2281! .- 100.1 Ayerage 99.89 15 PRESENTATION OF DATA AND DISCUSSION The following data represents the urinary sulfur analyses for six preschool children on four different diets: (l) basal diet. (2) basal diet plus fat. (3) basal diet plus starch. (h) basal diet plus sugar. The entire study comprised thirtyhfour three day periods; the basal diet included the first ten periods and the modified diets eight periods each. The presentation of data follows the above order in which the children ate the diets. beginning with the basal diet. Urinary sulfur Excretion on Basal Diet The average total sulfur excretions for the basal diet varied from h29 to #96 mg. (Table 6. Chart 1). The child I had the greatest aver- age excretion (h96.8 mg.) and also had the largest calculated intake (665 mg.). The child 6 excreted the smallest total amount of sulfur (h29 mg.) and had the smallest intake (559 mg.). Since each child received a constant diet. his sulfur intake should hays been uniform throughout the basal diet. However. there were vari- ations in the individual total sulfur excretions which would seem to indicate that the sulfur intakes were not constant from period to period (20) (Table 6). Hawks. Dye. and.Bray have shown by actual analysis that the nitrogen content of supposedly constant diets varied to some degree. 16 ; Tbtel sulfur chi-Might Intake p 0 A10 0flflfiu'fiufl.‘ E0 ) ('8 J o . be38 565s z; 0 s 0300303 ciu»-€ a»1e a» «use to p O A’s 17 17. '70 ' 17. 17.5 . 17.5 = 17.3! ) beret ion 7” e m Individual Average xv.) V'* \'.0 f’ 0' f ‘97 ) 7""— ~3.26 ~s.ss ‘0e9, .Iet‘ .w . - .- . -- ”-‘n-‘I. Char? I. 17 TOTAL 5ULFUR EXCRETION ON WL [JET / ‘ g 5 ' ’// Kat—M f’ \\\‘ / / \ 5/0 // \ ., \ \ I \ \- I \ 4»: _ ‘3 ‘34? (Girls) L 3 "1’: o , Y---- k K it /I\ a / \ ' " —-V’ 2 j— Z \\ '\\ 4" / \ \/ \ 4’1 i J 39 ’1? o 5 ii .6 18 Therefore. since nitrogen and sulfur are so closely associated in food sources. it seems plausible to assume that the above noted variations in sulfur excretion from period to period may have been due in part to variations in dietary sulfur. .A second reason for this theory is that the childrens' sulfur excretion curves followed the same general trend. therehy indicating similar inconstancies in dietary sulfur. For exp ample. during three of the periods (numbers 5. 9. and 10). all of the excretions fluctuated in a similar manner. and in three periods (num- bers 3. h. and 6). four of the six fluctuated in the same manner. The mg. per kilogram sulfur excretions were preportional to the mg. per kilogram intakes since the diets were computed and fed on the basis of kilogram bocy weight. Table 7 and Chart 2 show that the aver- age mg. per kilogram sulfur excretion was quite similar for all of the children. In fact. there was only 3.1 mgs. variation between the highest excretion (28.7 mg.) and the lowest (25.6 mg.). Although.c received the smallest amount of food and the second largest amount of sulfur in terms of mgs. per kilogram body weight. he had.the highest excretion on this basis. The largest child. E. received the most and excreted the second largest amount of sulfur and the girls both receiv- ed and excreted less sulfur per kilogram than did the boys. It is evi- dent. then. that the mere weight of the child.had no effect on his sulfur excretion-the determining factor being the dietary intake in terms of mgs. of sulfur per kilogram. It is interesting to note that. with one exception. all excretions decreased during the last two periods of the basal diet. This was without doubt due to the fact that while all of the children had gained weight by the end of the period. their | ----------- u o — - A . o . — .‘ - 0.. 0 ‘1'.-- o o - l v a - 4“. .e 1 c u “v e m - ~. v A~1 . ‘1’ . . — J "1. ' ~ 1 | I a...” o -‘ -~ ‘-.w- run-p . -‘r- o — l-vv- -- .- IV- a I . o- o — -- - ' ' ., 0 x - . . . - . . .» . ‘ r . ! ‘ 0 -. . J. . ll ' , .. a . C e 6 fi‘le.‘ 7 . p 0 :3 I l 9.. - ~ ~ -- -'~--. --‘-av aka-eo.‘ .-4-7. ¢ ’.— .._. - .9 w..-‘-.e-Co---Qr—e »-., ‘l 3 ' l :0 J . ' . . . 8. " q ' . . t . U 0 . a . ‘ '\ . l ' '. . . “ u..---9- Q I, ". . O nu . a. -J ( --° -- . I . ' ' i .. . t m. .e .' .2 0! u . . r“ S C J. “a J:- O in ' fie - o- - ”ou- - -.- has—s .-.~’---—. A - § ~v - “o.“e- '- O‘. 0 ".' - - .- - -.._‘- wow -'-0\ Q-u'f ‘ O .4 i a e. ..v-‘-- o e O - -- O O I .s I e I I ‘ o M \ n I n | I v 0 I ' I . l a. . U C .Q—e- excrefél Oberf 2.. SULFUR EXCRETION DER KILOGRAI‘? or BODY WEIGHT ON BASAL DIET 3] N [77/49. .5045” //l \ x 21 7" —— ’———" 2 / \ \ / \J 21 26.. £5 (504/3) 9;... 24" “+1.4. 23% 29 r=~~~ - g g / \\ // \\ 23 // \k.‘ \W \\ l/ \\ -——— \ - 21 \l \ v (Gk/5) J Md—-- Y—l—A—A. £31 2 5 4 diets were unchanged. thereby reducing their per kilogram food consump- tion. There was an individual period variation in the mg. per kilogram sulfur excretions. as was true of the total sulfur excretions (Table 7. Chart 2). But the general trends in variation on the basis of mg. per kilogram.excretion were more nearly the same than on the basis of total sulfur excretion. For example. the mg. per kilogram excretion curves of the two boys I and C were identical in their variations exp cept for periods h and 8. and the curve of 3 followed that of K except for periods 6 and 7. The curves of the girls. J. “J. and.Y. were simi- lar with the exception of periods M. 6. and 8 (Chart 2). It is inter- esting to note that in periods 3. 5. s 9. and 10. the variations for all of the children were in the same direction. and during periods 5. 6. and 7. the figures for four of them varied alike. In order that the degree of variation between children might be determined. each child's arerage sulfur excretion and his period variap tions from that average were computed. -Ae may be seen in.Table 7 and Chart 3. the child.K had the greatest percentage variation from his average excretion. ranging from -l3.lh to +6.20. while the child C had the least percentage variation. ranging from -3.83 to 46.27. The group average figures presented in Table 8 represent an aver- age of all of the children for each period. Under the circumstances. this kind of an average is permissible since all of the children re- ceived the same amount of food.per kilogram bodyweight. Table 8 and Chart h show that the individual excretion figures fellowed the period group averages very closely. Agaih. it seems logical to assume that the 22. ”y/ly. Su/fuf excreféa’ “-58“ w. <>D_>.:OZU Om. QCPWCD ”>0Dm3020, WED: EQSDCbr > 1 3c < Nu! chm... N§.+§Wb «\QKINngVL OQS tNWh \XW» a wwmlvr... ww+m~w1¢jifimwa+1 23 C/mrf 4. INDIVIDUAL VARIATI ON_ IN EXCRETION FPOI’I PERIOD GROUP AVERAGE. ON BASAL DIET Alon 2. firlos’s I Z \wxuxcso \\K\§h $\\ Rt 25 2h individual variations in sulfur output were due to a great extent to dietary sulfur inconstancies. TABLE 8 VARIATION OF URINARY SUI-FUR EXCRETION FROM PERIOD GROUP AVERAGE W Variation (8) Per. Group c K I NJ J I Average (mg/kg) 1 27.7 .5.u2 ---- +1.01 ---—- ---- .6.1h 2 27.8 +1.80 ..--- -l.80 -...- --..- ..--- 3 27.7 +3.61 -1.81 +h.33 +0.7h -h.33 -2.53 h 27.5 $1.00 +0.36 +5.82 «2.91 .3.27 -5.09 5 28.3 +3.53 +1.u1 +5.30 +3.18 ~5.30 ~7-77 6 27.9 -l.08 i0 +8.2u +0.72 +0.36 -9.32 7 27.7 e3-97 +5.05 +5.05 +3.97 -5.78 -11.55 8 28.1 +8.5h -1.h2 +2.85 -0.71 -3.56 -u.98 9 26.9 45.20 41.h9 +3.72 +2.60 -h.83 -8.92 10 25.2 11.11 -5.55 o0.h0 +2.38 -3.97 -5.15 Table 9 presents the individual average percentages of dietary sulfur excreted in the urine. The averages varied from 70.0% to 76.1% and were obtained by using the calmlat‘eaakotal daily sulfur intake and the average of the daily total sulfur excretion for the entire basal period of each child. Although these figures are inaccurate to the extent that the actual food analysis figures were not used. they were very similar to those presented by Hunscher. Macy. and Cape.(5) who reported 76.h3 and 76.26% excretions of dietary sulfur by two chil- dren 11 and 7% years old respectively. on a normal diet over - 5 experimental periods of 5 days each. The percentage of dietary sulfur excreted in the urine apparently had.an inverse relationship with the percentage gain in weight (Table 9). For example. the child C excreted the highest percentage of his TABLE 9 CHANGES IN WEIGHT AND PERCENT OF SULFUR EXCRETED 0N BASIL DIET Child Change in Weight g:;;::y (kg.) ($I Excreted 0 +0.01 0.07 7g?% 3 +0.25 1.h3 7h.8 I +0.32 1097 7309 J ¢Oe38 2023 7203 MJ +0.3h 2.1h 73.6 I ‘0e32 1.98 70e1 dietary sulfur and made the least percentage gain in weight while the child.! excreted the lowest percentage of her dietary sulfur and made the second highest percentage gain in weight. Hawks. Dye. and Bray (21) have shown that with an increase in diet nitrogen. there was an attendant increase in retention of nitrogen and also a greater gain in weight. Therefore. because of the close relationship of sulfur and.nitrogen. it might be expected that there would be a correlation between sulfur retention and gain in weight also. Although this paper does not present retention figures. it is interesting to note that the children who made the lowest percentage gains in weight (K. C. and I) excreted the highest percentage of their sulfur intakes on the basal. diet and therefore may have retained the least sulfur. Since the greater part of ingested sulfur is contained in protein foods. the sulfur excretion would be expected to parallel that of nitrogen. Several investigators have noted this parallelism and have expressed it as a.N/8 ratio. Therefore. for purposes of comparison. the N/S ratio of each of the experimental sugjects was determined and presented in Table 10 and Chart 5. 'The averages of the individual ratios for the entire basal diet ranged from 15.02 to 15.53. Although the average N/S ratios of the children on the basal diet were very nearly the same. each child showed variations from period to period. And again. as was true of the sulfur excretions. these ratios fluc- tuated in very much the same manner. 27 Hm.ma mo.ma nm.ma n:.ma mm.ma -.ma .a4 mm.ma ammm aH.mH amam -.ea maam mo.ma :mam --- -- H:.:H mmom oa mm.ma mafia m~.:a mamm m:.ma mmmm Hm.ma mafia ~m.ma amoa. as.ma mmmm m ma.ma moo~ :m.ma mmem mo.ma Noam o~.:a mmea mm.ea maam ne.:a «mam m mm.:a Hmam mm.:a mmmm om.:a oemm o:.ma omma e~.:e anoa mm.:a momm a ma.m~ mmmm mm.:a ammm mm.ma onmm mm.ma omom Hz.ma mmoa ~H.ma mamm m Hm.ma mmmm mm.ma mmoa mm.me mega ~m.ma amma om.ma anaa mm.ma mamm m :m.ma mama mm.ma mmo~ am.ma ammm mo.ma omam mm.ma mafia :m.ma aoam : mm.ma mama oo.ma mmam Hm.:a mmmm em.ma mama ea.ma mamm ma.ma mamm n --- mmam --- -- --- mmzm mm.ma came --- -- mm.ma ommm N m:.ma mamm --- -- --- -- mm.ma mama --- -- -.ma ammm H “.mam “.mav “.msv A.esvl A.mav A.esm deaeaaa dopeuan deeeaoxn do» shown doaeaaflfl deaeuonn cause 2 chasm a cause a cause a chasm a canoe n m? as a. a? :33. a; dance Q: H38 a? 1» 8 a} 129 .2.” a e a: a a o I I. fig .33 no 23 {a CH “an. Char?" 5. N/5 RATIO ON BASAL DIET /7 /6. 15 l3 I7 [I R ‘ IV/S ref/e % x U \ N (N 4. [- OP I) ah 6 gr} 04$ URINARY SULIUB.EXCRETION ON HIGH.FAT DIET Previous studies completed in this laboratory demonstrated the necessity for an adjustment period following a change in diet. The investigators found that when the diet nitrogen was increased there was a corresponding increase in nitrogen elimination. The diet change caused a fluctuation in urinary nitrogen whichupersisted over a period of several days until a state of equilibrium was established. This occurred at the end of about three periods (nine days) following the change in diet. While the modifications in the present study did not involve a change in the mineral content of the diet comparable to the above study. it seemed advisable to reserve the first nine days of each diet phase as an adjustment period. During the high fat diet the individual average total sulfur 01h cretions of the adjustment periods (numbers 11-13) were very similar to those of the experimental periods (numbers lh-lS) as may be seen .‘in Table 11. For example. following the adjustment period. the aver- age excretions of’J and-& decreased only 0.3 mg. and 2.3 mg. respec- tively while those of NJ and 0 increased h.6 mg. and 6.2 mg. respec- tively. The greatest change was that of E. whose average excretion increased 13.5 mg. following the adjustment period. It is of interest to note that the two children J and.I whpse excretions varied the least had had no change in diet. except ton the additional fat. And‘the child 1. whose average excretions differed most widely. had had the greatest 3O HE J I hr. Weight Intake Excretion Var. Ir- [Individual ’Lverege m.) (-2.) Inc-J ? fi 11 15.11 i 18 15.19 I 1: 15.31 ‘7, is. ‘ 1‘ 150“ 15 15.58 16 15s“ 11 18.71 is i 4 A's 11 18 18 AV. 1‘ 16 17 10 Av. since ea... -—e ~~~.-'- .“ -‘. -Vo- - on ”w .'-O-‘u -n'w- .-Q -. e <~ C —owo—.—- .- *--‘-~. ’9 I tentI ‘ I ..- s.- MS 0‘ V a. I” e X 1 ..... e I .I. o. 3. .- .,-.. .J . . '1, .1-.. . . ~ . h . v—O- b -"r -9- '9‘;- .‘Q".-COO"”-- perm child 1 Ir 09,! '0'.r1’ VI... ‘1 III- 'I- I- I ll.’ . n .. o . ... i L, N! m m e e . . O ill ‘ J lleJ . .4 .. urn-.- I lost hrs ' 11. c an. 01“ (J child thas tin lug . hfl'g - ---...—“ Great- 31 change in diet. However. none of these differences is significant since they come within the limits of experimental error. The total sulfur excretions did fluctuate to a much greater ex-- tent during the adjustment period than during the following periods. as may be seen in Chart 6. These fluctuations indicate that there was some disturbance in the sulfur metabolism and that the adjustment period was therefore advisable in this study. The individual average total urinary sulfur excretions of the children on the high fat diet varied in preportion to the sulfur in- take (Table 11). For example. the children n and I received the 1dr- gest diets (diet factor .95) and excreted the largest amounts of sul- fur-$52.5 and 1138.1 mg. respectively. The child 0 received the smal- Iest diet (diet factor .82) and excreted the least sulfur—397.0 mg. The period to period variations in total sulfur excretion were more nearly alike on the high fat diet than on the basal diet (Table 11. Chart 6). In four of the five periods. the excretions of four of the children varied in the same manner. The individual average total sulfur excretions were all less on the high fat than the basal diet. which may have been due to one of. or a combination of several causes. First. the diet factors of the children I and J were unchanged. while those of C. E. and NJ were less. thus decreasing the sulfur intakes. That this may have been the cause- tive factor in the decreased sulfur output was substantiated by the fact that I who had no change in her diet had less difference between her averages on the basal and high fat diets (5.6 me.) while E had the greatest change in his diet and also had the greatest‘difference Chorf' @- TOTAL 5ULFUP EXCRETION 470 ON HIGH FAT DIET +70 ‘3, 4501 i; In t / .. 430—— — II L \ a 410 \| \ 1‘ ’l \ / \\\ 3 / \\ // . 3 a _ — . t“ /// \\ / (Kn-Is) . / n3--.“ / VMA— W1: 12. /W 32 33 between his average excretions (“.3 mg.). Secondly. all of the children gained weight more rapidly on the high fat diet. 'hile the highest percentage gain in weight on the basal diet was 3*. the lowest and highest gains in weight on the high fat diet were 5.16 and 7.28% respectively (Table 12). Thus the more rapid rate of growth may have caused a greater retention of sulfur; T1313! 12 MARION BETWEEN CHANGES IN WIGHT AND PER CUT SUI-m EXCRETION ON HIGH TAT DIE}.I I Change Dietary In Sulfur Child Ieizht lxcreted Ore.) (e (5) C e0.“ 5.89 71.8 I +0.91 5.16 70.8 NJ +1.18 7. 28 69.1 J 01.011 5.92 65.8 1 +1.21 7.11 68.1 AV. 69.3 The individual average mg. per kilogremexcretions on the high fat diet varied from 22.8 mg. (J) to 25.”- mg. (C) (Table 13. Chart 7). Thus there was only 2.9 nu. difference between the highest and lowest average excretions—even less than on the basal diet. The calculated average intake varied but 1.1 mg. from the highest to the lowest. which was also less than that noted on the basal diet. Again. the child C received the smallest total amount of food but actually re- ceived more sulfur per kilogram and excreted more on this basis than .-. H *--Oebo‘ .~-.-. .0 a. u so. w~o -..OO.. L... .0-- '. I. l U Mo—a‘-.. ‘ O~.~«, ,_ '-_ ,...r -. .a»--.u--.‘-v-"’-""‘ ‘,, ... , . . .,_.o.—'—' e -v u-or« —-.-.’0~ 0 I a " " ,o a- . ——-o o 0 ’v V ‘ ‘ “ . . ., - . -. _... _I~ ' - _.-- . -..J . - - .-Q Q o O y- oa—CW'A’ee . eon->0 o-o‘tF-W e- pa . an-.- o . -- - a e — . U s s I. I I 3! ' ‘ ‘ I L. i: I ' O ' . . _-' ' ._ r. .g... 09".. 3-1 ’rq-‘ . -o o u.-. -.' o 0:- O »-. - . .- --a - r I » . ° 0 . ' ' ! t . ‘ pr. . I J. _ ‘ 9 l A O . e '1 ~-- ' -- - 3 ‘ ’ e] I - l . a U :9 ' , i i ‘ ‘ 9' “ - _ I 0 a C 5v 5 s I 0 V i . ° 2' 4 . ‘ .9 . u . ‘ . . _ . - c c o v - - o - ~ 0 A. vs- -5 0‘-“ “‘1‘ ‘ ‘ P- .-‘.L -- ’ | g a H ‘ i ' ‘ . v .v' ‘ .. O ' ‘ . I z' w , .. ‘a . . C c ' ' ' I ' g ‘ g. . a I f e l . I. ‘ . ' ‘ LIV .**-o 0-: . I I ~ . s 1 a e . r o ‘ ~ ‘ 7 l -. q ( o. ' d ‘ D --‘----0Qo-~*. ,. -.—.—o. a. ‘0 "i-th-e'.»d.v.-ou ’ D Q I ' J ‘ . 0? ‘ -1. ‘ O 4 O . o O o . .e 0 s ’ ’ s ; e ‘ ‘1‘ ~ I . ' I L 9 ‘ 1 ., ' ‘.l .- ‘Owa.-!-I.- s l -. a. o ..w... on- ~ a. --._,,'._ Chg/f 7. sou-up acumen pm KILOGRAM OF BODY WEIGHT 0N HIGH FAT DU I 'V *2 ° / ' / s?“ /- gs / h 21-» I/ 9‘ / h' 123 -—--/l / l‘ / 22 2L" Ila. 1.3 “mods na---- Y—a—A—A— \\ /\\ \ __\____./.. a. \ / \ / J. n [a 36 did the other children. It is interesting to note that on this diet as on the basal. all excretions but one decreased in the last period of the diet. Again. this was no doubt due to the fact that the in- crease in weight caused a corresponding decreased intake per kilogram. There was an individual period to period variation in the mg. per kilogram excretions as mabee seen in Table 13 and Chart 7. How» ever. the fluctuations were more uniform than those on the basal diet. For example. in periods In. 16. and 18. four of the five excretions varied in the same direction while in period 17 there was practically no change in four of the excretions. These figures again emphasise ‘the probability of a common dietary variation. The period to period variations from the individual averages were less than on the basal diet as shown in Table 13 and Chart 8. For example. MJ had the greatest percentage range from her indivi- dual average-from -6.75 to +8.86. while C had the leasto-from -3.15 to .2.36. Ihe excretions of all the children except 0 and.J showed closer conformation with the group period averages than with the individual diet averages (Table 1h, Chart 9). In other words, each child except C and.J had a greater percentage range in variation above and below his individual average than above and below the group average. The average percentages of dietary sulfur excreted on the high fat 41.1 varied from 65.8 to 71.8% (Table 12). Thus. although the children received less total sulfur. they apparently retained a greater percentage of that ingested. since on the basal diet the lowest per- centage excreted was 70.0%. It is interesting to note here also that 37 W‘QwKQ C 5‘ h Q\ ex . r. L. h. J .xhh a. .h 1 h. 2 2... 5 4w. /4 /.M.N.W nth n/ WI! III F rflr Ill] 1. J 9 x in” fl u o n /‘I/ .Jnu. / //|IIIII.|\\IWN PEG F< 4.420352. ZOQL nZoFquxu «5.....50 LO WZO:<_Q<> TABLE 1h INDIVIDUIL VARIATION OF URINARY SULFUR FROM PERIOD GROUP AIRBAGE ON HIGH FAT DIET i Variation ($) Per. Group I C 3 NJ J Y Av. A L_ (mg./kg.) 1h 2n.7 .o.ho +5.67 +u.u5 -h.h5 -h.h5 15 2u.6 +5.28 +3.66 -2.85 -6.91 +1.63 16 23.7 +9.70 +2.95 -6.75 +6.33 £0 17 23.9 +7.95 +2.09 -o.h2 -6.69 -3.35 18 23.2 +6.90 -o.86 -1.29 to -u.7u 38 the two children who had no change in diet (J and.Y) also excreted the least percentage of their dietary sulfur. Again there was a correlation between gain in weight and per cent of dietary sulfur excreted. The two children C and I (both boys) made the least per cent gains in weight (5.89 and 5.16%.res- pectively) and they also excreted the greatest per cent of their diet- ary sulfur (71.8 and 70.8%, respectively). The individual average N/S ratios were quite similar. varying only from 15.0 to 15.15 (Table 15). Four of the five children showed a slight decrease in their individual average N/S ratios from those on the preceding basal diet. but the decrease was too small to be siga nificant. .As seen in Chart 10. the N/S ratios followed the same general The boys' curves (C and.E) were almost identical while the trends. girls"were less similar. 39 Cirorf' 9. WLZDIVIDUAL VARIATION IN EXCRETION 27 PERIOD GROUP AVERAGE 0N HIGII FAT DIET E 3 a U as El 1’25- 2- a 3 $ A A a a "< ° ‘23” m—C k\ e 8 A-E 6‘ 6 x . x‘d 22/ I I e—HIJ Hui-£073 /s 18 1‘7 78 . ’Y Cfiarf IO. N/s RATIO ON r-uen FAT DIET 8.3 3.3 8.3 3.3 3.3 .84 $.3 $3 3.3 9.8 3.3 RS mm.3 #8 mm .3 Row 3 2.3 38 3.3 $3 8.1: 3.6 .33 Ram 3.3 3% 3 3...: 3mm R}: 8% 3.3 nmmm 03: $8 mm...” 98 3 d3: 33 8.3 ammo 8.3 mos 8.3 an: was: $8 3 $3 38 $3 mane «:3 38m $1: Ram 8.3 man an ax: 3.1: :31: 8.3 3.3 .5 max: 83 Sr: 9% mm .1: mmmm :3: «Gm 2.1: $3 3 3: 83 3.1: mnmm am .1: 33 3.: same 3.3 $3 3 8.3 3% 8.3 3R. 3.3 83 2. .1: gnaw 93: 3mm . S “83 “.an Ads 1an A83 coaches” deaeaufin eeaoaofln .0333" eeueaan , 3.3 a 333 z Show a £3.— a Spam 3 a} 38. {a .33 a? 389 a? 333. a? 38 .8." u h. a. a u an as. man 3 85 a} 3 can hl URINARY sunrus.sxcnsm10n ON HIGH STARCH DIET The remainder of this study will include the excretion figures of only three children (Y. C, and.E) inasmuch as the other two (MJ and.J) failed to consume the diets quantitatively in several of the periods. The individual average total sulfur excretions of the adjustment periods (numbers 19-21) varied little from those of the periods fol- lowing.(numbers 22.26) as shown in table 16. During the latter periods the average excretions of O and.Y increased 6.0 mg. and.13.6 mg. res- pectively. while that of’E increased 20.9 mg. Although these differ- ences were not large enough to be significant, they were all greater than on the preceding diet. It is also interesting to note that the same child (0) had the largest variation between his average excre- tions for the adjustment periods on both this and the high fat diet. this fact probably indicates that the change in diet produced a dif- ferent effect on the sulfur metabolisms of the individual children. The total sulfur excretions fluctuated.to no greater extent dur- ing the adjustment periods than the following periods (Chart 11). From this chart it would seem that the adjustment period probably was not necessary on the high starch diet since the cornstarch seemed to cause little. if any, disturbance to the sulfur metabolism of any of the children. On the high starch diets. as was true on the basal and.high fat C barf //. TOTAL SULFUR EXCRETION ON :1th STARCH DIET ‘u 3 3 0 h x 4-70 ll L $450 5'. . 4 ._. J __ Q» ‘~‘ . 4’ (Girl) Y—A—JLA— /¢ 240 , 2/ 2L2. 2'3 2: 25 26 “/7045 he '6 Child hrs 19 21 unuua AVe 19 A's wanna.» 21 AVe T —a .\ I a. t z A. he fl. 0"?! l 0 . .o .. -e . A a.“ .J. u y. o. u... .- . .. . ~o . ‘ . .. a Oi}-.- e u .._ a. .1 . e .L - . h a . q .\ wx'll...‘l.en|r iv _ .s e o . o . h ‘ sv- In . O n, e a. s: .l a'svales ud-‘l.‘\ o . - u 0 .1. .n . o .u . o .. . o r. e.---. -—---. “a. tlt‘u‘w't‘ 3. ;VI! . -0 "- ..,I‘. O .... H I .u ‘1'. ._-- _ <-" -Q—h-.“.‘n s t . o D I .....'. .9- J.— 0. £ e ev . - . .. .em . w .0 J a l . e a I... O -‘ I -I A 0 o 3". I - Illit - '1..- - '.‘.". '1': ‘*-L‘l s . -wu--w"m—— >s- . diets. the average urinary total sulfur excretions varied in accordance with the dietary intakes (Table 16). Again the two children s and I received the larger amounts of food. (diet factor .95). and excreted the largest amounts of sulfur (326.” mg. and h31.l mg. respectively). The child C had the smaller daily food intake. (diet factor .82). and excreted the least sulfur. h05.8 mg. The period to period variations in total sulfur excretions were quite similar (Chart 11). While all of the excretions untied in the same direction in only three of the five diet periods (22. 2M. and 26). the general trends of the individual excretions were more uniform than on the preceding diets. The average total sulfur excretions of the two children I and.Y were less on the high starch than on the high fat diet (26.1 mg. and 7.0 mg.. respectively). The excretion of child C (h05.8 mg.) showed a slight increase of 8.8 mg. over that of the high fat period. All variations were too small. however. to be significant. The individual average excretions. without exception. varied less from the high fat to high starch than from the basal to high fat diets. This was pos- sibly due to the fact that the average sulfur intakes varied less from the high fat to high starch than from the basal to the high fat diet. or to the fact that additional calories produced the same changes whether they came from fat or starch. The individual average mg. per kilogram urinary sulfur excretions during this diet varied from 21.9 mg. (I) to 2h.7 mg. (C). as seen in Table 17 and Chart 12. Thus these values were progressively lower dur- ing the basal. high fat. and.high starch diets. as were the dietary I‘lli 1w .. 4‘ . w mTlIo It . i . . w an H .w. .e. ‘0 . w e U wv m 2 . a .. ... .. a. . . . . . a e. .1 , a. ” um. 31. d; “ v. . C re n a .W .. .. .. . , . Io... .. ate“... t. - ~’-O U..." I e . .. , V I J. I 3‘ . I‘. fi 0 '1 . 1"." s. A7. Per; LVe a o . .a o ‘ .. a L . s e h .s. s . i I . - a. L x. a .a . e .- e . .. . L o . . . s . I .s e . .. 0 lb . I v I x. . ”r I. fV .t. w c . . c . . w 0 . .. e .11" 0 O o I '0' DIP. 1.. ...'I'D-: I. -‘i‘:‘-i'iute 'i‘. ‘1'.’ .a. a m .. n . w o .. a w. . .. . w . . M e r a a . . . ti u e . . . .wl’ - i’ O. 0“ a! .Q-. OJ- 3:.:'.’ ‘-‘I- ‘l."-l‘.' C 1.. .‘ ’Isll. O ‘ ,.s. -s! " I. - . . a —“ . ... . . . . “ e e a a .. .I . J I r . . s . .s. . ._ s « .. . ~ . . . 4. . t .e e .o r \— . . .. s a . _ a ... e e e e e e e e e s . . ~ 0 . V- u‘ “0‘ V i \ .V.‘ M. OI... ” a. m a b u \ .l e . . h I} .et K e. l ii‘w,‘,“ )ei’s’ .. .' 'l '.‘."0' - ‘C" .1-" .l‘ 10“... O . . n o . A . . . ~ . . .1} y o a c e . c a .. .. D l w L . s M. . . a.» , . e e e o a p e e e e e .. . . e a. . . . . o. . .- p-g .. a ..-O s . .- .- . M7. sulfur excre’féd Chart“ IZ . SULFUR EXCRETION Pin KILOGRAM 0F DODY WEIGHT ON man STARCH DIET it: N \ I ha I I l I I§ it h} r ’zt‘ 2/ \ ‘0 I N \ N n N W ’0 st h‘ 0) 26 __~_hg£_-L__‘____g ~_ ; __; H—ZL—%~—'__#LW_—_‘—— _ h7 sulfur intakes on the basis of mg. per kilogram. Although the calcu- lated average mg. per kilogram intakes varied only 0.1 mg. from the lowest to the highest. the difference in excretions was 2.8 mg. This figure was practically the same as the difference on the high fat diet of 2.9 mg. The child C again excreted more sulfur per kilogram than the other children. even though he received the same amount of sulfur per kilogram. In contrast to the other diet figures. all excretions increased during the last period of the high starch diet. It seems probable that this was caused by a fluctuation in dietary sulfur rather than any disturbance to mineral metabolism. The same similarity between individual mg. per kilogram excretion curves evident in the preceding two diets was true of this diet also (Table 17. Chart 12). All of the childrens' excretion curves varied in the same direction in three of the five periods. The boys' curves did not show the marked similarity whichihey had on the other diets. however. The mg. per kilogram excretions from period to period fluctuated less above and below the individual averages than oh the two preceding diets (Table 17, Chart 13). for example. the greatest range in varia» tion was that of Y («n-6.85 to -S.O2) while the smallest was that of C (.2.n3 to -2.02). Two of the individual mg. per kilogram excretions (E and.I) again varied less from the group average than from their individual averages (Table 13. Chart in). Subject 0 had a total range in variation of h.u5 from his individual average and n.70 from the group average. a difference which is not significant. Char-f 13. VARIATIONS 0F SULFUR EXCRETIONS FRO” INDIVIDUAL AVERAGES IN HIGH STARCI'I DIET 27 25" A / .___———/ V 23~ 3 C \ / B V u 2/" Au—247 . E I? I ”'7) (,4. _ are a...) k :1 a) 2 / k /\ a: V :2/ 2 (flu-Z/.4mq.) [7. l l 4 I ’ . 25 36 £2. 23 S“ ‘ Quads Charf’ If. INDIVIDUAL WRIATION IN EXCRETION ER 0” PERIOD GROUP AVERAC E 0" HIGH STARCI'I DIET Q 27 5 25. a ‘3‘ ‘51 a ‘3 m E 23L. W t - c. t :2” ‘3 13-0 2 43-5 22 £3 21? - 25 :6 firiods TABLE 18 “9 INDIVIDUAL VARIATION OF URINARY SULFUR FROM PERIOD GROUP AVERAGE 0N HIGH STARCH DIET Variation (6) Per. Group C E I Av.‘ (ma/kg.) 22 23.0 +5.22 .o.hu -5.22 23 22.2 e9.92 -3.61 -6.33 2h 22.9 +9.62 ~5.68 -3.h9 25 22.7 +7.07 -2.6u -h.35 26 2n.o +5.h1 -3.33 .2.50 The average percentage of dietary sulfur excreted.varied from 67.6% to 75.5% (Table 19) which were lower values than on the other diets. Since each of the children received less sulfur per kilogram on this than on the other diets. it would appear that with one excep- tion (C). they retained a greater amount of the sulfur ingested. TABLE 19 RELATION BETWEEN CHANGES IN WEIGHT AND PERCENT SULFUR EXCRETION 0N HIGH STARCH DIET w Change Dietary Sulfur Child Weight Excreted Ute.) (i) (73) c 90.68 .h.26 75.5 E ‘0e72 ‘3e88 6709 r .o.95 45.05 67.6 50 Each child received a calculated 20% increase in calories over the basal diet. Therefore we might have expected a gain in weight compar- able to the last diet. However. there was a smaller gain in weight in all cases with an accompanying increase in sulfur excretion of only child C. E and‘! had slight decreases in their percentages of dietary sulfur excreted. The child.Y had the greatest percentage gain in weight and the lowest sulfur excretion on the basis of percentage of sulfur ingested. If the actual calories were the same on both diets, the preceding conclusion that greater retention of sulfur paralleled greater gains in weight was erroneous. The average N/S ratios varied from lh.90 (C) to 15.05 (B). These ratios were slightly lower than on the high fat diet. but not signi- ficantly so. The ratios were also far less variable on this than on the preceding diets. as may be seen in Chart 15 and Tables 20 and 21. For example. child'! had the greatest range in variation from her lowest to highest N/S ratio (1h.70 to 15.hl). which was less than half as great as her range on the high fat diet (1h.ll to 15.88). From the foregoing figures we might conclude that the apparent greater uniformity in sulfur excretion on the high starch diet was due to one of two causes. First, there may have been less variation in dietary sulfur during the high starch diet. Second. the additional calories in the form of cornstarch may have been less disturbing to the sulfur metabolisms of the children than the additional fat. 5]. Che rf‘ [5" N/S DAT :0 o N HIGH ST ADC n D [ET H I? l 20 2 Re 'I l riot/s 2t 23 .1 Z4- 4* 25 2'6 TABLE 20 3/: mm on men season um 1 Per. Total N/S Total 'N/ 8 Total 3/8 x Ratio 1! Ratio 1! Ratio lmcreted lsereted Increted he.) has.) (ms.) 19 6191; 15.02 6761 1h.85 661I3 16.52 20 57:5 1h.95 61120 114.38 , 6093 11am 21 5896 1h.6h 6621 15.014 6h81 11L.82 Av. 1h.87 114.76 15.36 22 6010 15.22 6507 1h.96 6288 1h.87 23 5865 1h.69 6196 15.16 6103 1u.99 2t 6099 114.73 6313 15.31 6&23 1h.7o 25 6075 15.10 61428 15.07 6558 15.h1 26 6169 1h.75 6612 111.77 6868 flash Av. 1h.9o 15.05 1h.96 was 21 was Is newton or 3/3 RATIOS or men season 11m Child Av. Highest Lowest Range 0 15. 22 1u.69 0.53 s 15.31 111.77 0.51; 10' 15.71 1min 1.30 Y 15.h1 1h.]o 0.71 _ 52 53 URINARY SULFUR EXCRETION ON HIGH SUGAR DIET The individual average total sulfur excretions of the adJust- ment periods (numbers 27-29) differed little from those of the follow- ing diet periods (numbers 30-3h) as may be seen in Table 22. The average excretions of C and.Y showed an increase of 2.5 mg. and 2N.2 mg. respectively while that of E decreased 9.1 mg. The high sugar diet varied from the others in that all of the childrens' excretions varied in the same direction in one of the adjust- ment periods (27). while they were not all alike in any other period of the diet. This points to individual differences in sulfur metabo- lism rather than common dietary inconstancies. Again the individual average urinary sulfur excretions varied with the dietary intakes. I had the greatest average excretion (M72.h mg.) and also had the highest average intake (705.0 mg.) as seen in Table 22. The child C excreted the least total sulfur (h20.h mg.) and had the smallest intake (592.0 mg.). All of the children had a larger total sulfur intake and a larger excretion on this than any of the other diets. except the two boys C and.E who had greater excretions of sulfur on the basal diet. The individual period to period variations in total sulfur elimi- nation were not so similar as on the previous diets. In no period were all excretions in the same direction. The boys. however. did show a close similarity in their excretions (Chart 16). "e 81 58 A's 54 o..g-v" e- -1. ‘Q—e --.—.w ‘O H . It!“ 0 -LIIOI. e L. m 9‘- V! H .,a .. .. fl .. . 1 t. l\ I.» H." 11. c . 7 C O D "l‘ -‘n‘.-+'i,‘t II‘ I ‘1'. '."s C -.'.. .ul' i.. "\'u .‘U. I c. .\ C p u. is I .0. -II..'.' '.’ I ’U‘I I. . ',‘IC 0 o- ‘ M- 5..-..- o-Qv..--.~ .‘fi. 55 Char f /6. TOTAL. SULFUD EXCRETION ON I'IIGI'I SUGAR DIET 490 I /\ / 47o- / \\ / \ / / \ / / \ / 4x— / \—"'—-"=‘-‘- ————— J— / \/// / 430.. 410 /\f (50’s) 3 .. C m E____ 5/0 M7. Sulfa/- 'cxcrcfid Art'ods 56 The highest average mg. per kilogram sulfur excretion was 2h.6 mg. (C) and the lowest was 22.6 mg. (I) as seen in Chart 17 and Table 23. C also had the highest average sulfur intake (3h.9 In8.) while E had the lowest (3h.} mg.) on the basis of mg. per kilogram. The 1.8 mg. variation between individual average excretions from lowest to highest was less on this than on any other diet of the study. The calculated dietary sulfur intake per kilogram varied only 0.6 mg. from the lowest to the highest. The individual period to period variations were quite similar for the boys C and.E. The excretion curve of the girl. I. however. showed little agreement with the other two. The fact that she was gaining weight more rapidly may have tended to affect the regularity of her sulfur metabolism. The mg. per kilogram excretions varied above and below the indi- vidual averages on the high sugar diet less than on any of the preced- ing diets (Table 23. Chart 18). For example. the values for I varied the least from his average. -l.33 to 01.77%. I again had the greatest variation ranging from -3.51 to 03.51%. Both E and.Y showed a markedly smaller range in variation from their individual average excretions on the high sugar diet. C had practically the same range as he had on the high starch diet. Table 2M and Chart 19 show the individual variation from the period group averages. E and I both had greater.ranges in variation from their individual averages than from the group averages. In this as well as the other diets C had a greater range in variation from the group average than from his individual average. Evidently if there 57 Char 7" /7. SULFUR EXCRETION DER KILDGPAI‘I OE BODY WEIGHT ON HIGH SUGAR DIET ‘6 // \ /7 It: \’ 03°”) E“ «J / K 22a- 0 U k _ _ _. - U 3 #____________IL__% 1\ 3 24 v 3‘ :2. _J_ 2 (Girl) 22" YM— ZI. __| 1'? Z 80 3/ 87. 33 34' grinds O - .. l C I ‘ o . - .- .- ~. ._ -.- . ”a... o - o. - - - ~. . - ..] . o o.. -- . s.-ew.-o-~"”-~“-w - 0- "" c r " " -" r’-’- “'"‘ ”T "' ' " .~. ..~ -u.-.....w--—.a -v‘—" I s-«;., . . - .. . .v-o . on . -. - s .s-el‘ - . .-aw-’ A - - ‘0 “' ‘ "'I“'. h ’ - "t‘ " - " .. . Ii 0 I - 0 ,' e . I ' g .. "4 '. L VJ ob) ’ Q, o . 5.5 - I . -~o-a-....v--'sro-¢‘ I “"""' "‘"""" " ' T, I ' . | ._ ' ‘ " ‘.. I ' ' A u h~' t ‘ _‘ -J l . - , . l - . . U . v 3 ' r O C °t C I s C .' -9 0.- - . o ». - d~--- - - e O I \u- ~.~ -- ’ . I 0 Q . . n 0 . o 7 v C I | ‘ O r - 'I"s e -"-‘*‘ ‘u. . I I ' . u ‘ ' a“- s .3. ' . ‘ 3' f l I I '.I n." a a . I I I t -..' y - . I Q of». s ' g . , ‘ . e o . ‘. s <-.—. o ‘ C ’ r "U h' 0 ‘ .L'.’ I I v ' ‘ ' C s , v. ' . ; a}! ‘ ' . I I ': IO. ‘ ~ ’.D 7 I e , ' , ‘ .. ‘ 0.3.' C . :‘I 9 t O ' I O . ‘ 1 r V."’ .1: - _. . -0‘--.-~-“---o-n - . . ---‘.-o 59 Char f‘ l5. VARIATION5 DI’ SULFUR CXCPETION5 FROM INDIVIDUAL AVERAGES 0N HIGH SUGAR DET 27 Z5 /\ H E. (AV-‘24“ ”’7') (Av.—- 22.6 0”.) N x l C) Su/fur ea 6’8734 I: Z3L/\ s‘: \.z/ .— n Y ‘C (A .r. —. 22.5 my) ”be 3': 3'; *3 31? 3.0 5'! 3‘2. 33 :55} “dads Charf' lq. INDIVIDUAL VARIATION IN EXCRETION FPOI’I PERIOD GROUD AVERAGE 0N HIGH SUGAR DIET E 27 3 25" E! a a a G k . 3 A A A ,9 {a - 3:3 k 2 «Y I? 3:: 37 3‘2. 3‘3 3‘} 60 were any dietary variations causing the fluctuarions in excretion, C was less affected by them than were the other children. TABLE 2” INDIVIDUIL VARIATION OF URINARY SULFUR.IROM PERIOD GROUP AVERAGE 0N HIGH SUGAR.DIET Variation ($) Per. Group C E I Aw. (mg./kg.) 30 23.1 95.63 -3.h6 -1.73 31 2;.n e2.56 -2.98 .0.86 32 23.3 .5.58 -3.u2 -1.71 33 23.1 98.68 -3.u6 -u.77 3” 23.9 23.3” ~3.76 --- The average percentages of dietary sulfur excreted ranged from 65.3$'(B) to 70.5% (I) (Table 25). Thus the average of each child was lower on this than on any of the preceding diets. Since the vari- ations from both individual and.group averages were also less on this diet. it would appear that the sulfur content of the high sugar diet was more constant. Therefore, the children retained a greater amount of sulfur. There was a correlation between change in weight and amount of sulfur excreted.as on the other diets (Table 25). 0 made the least percentage gain in weight (h.18$) and.also excreted the greatest para centage of his dietary sulfur (70.5%). Y made the greatest percentage gain in weight (6.28%) and excreted the second.smallest percentage of 61 her dietary sulfur (66.5%). earns 25 RELATION BETWEEN CHANGES IN HEIGHT AND PERCENT SULFUR.EXORETION ON HIGH SUGAB.DIlT Change Dietary In Sulfur Child Weight Exereted Org.) (7%) (75) c 60.69 .n.1s 70.5 e0.87 eh.52 65.3 I .1.2h $6.28 66.5 The average N/S ratios ranged from lh.73 (C) to 15.13 (I)(Table 26. Chart 20). These averages of C and.Y were lower than they had been in any of the preceding diets. E had the same average ratio on this and.the high fat diets-~his lowest ratio was on the high starch diet. um: 26 8/8 mm 011 HIGH SUGAR 11m 1mm 11: N/S mms ON .1" c s I Per. Total I u/s Total N/s Total we 1: Ratio 1! Ratio ll Ratio Excreted lacreted Imre ted (32.) he.) he.) 27 5900.0 1u.67 6936.0 15.78 6692.0 15.2.1. 28 6311.0 111.95 6862.0 111.81; 6960.0 15.17 29 6289.0 111.69 66911.0 13.93 6705.0 1h.82 Av. 1u.77 114.85 15.11; 30 6211.0 11:39 66811.0 15.12 6936.0 1n.92 31 6170.0 15.10 6826.0 15.15 6989.0 1a.}? 32 6216.0 1m; 69110.0 15.112 7216.0 15.15 33 611h.0 111.17 6803.0 15.16 6783.0 111.70 3h 62111.0 111.67 6853.0 1h. 78 ........ Aw. 111.73 15.13 111.79 mm 27 62 HIGH SUGAR DIM Child Highest Lowest Range ’ Ratio htio c g 15.10 111.17 0.93 I 15.15 111.37 0.78 53 Char f to. N/5 RATIO 0N men SUGAR DIET / /\ /5}\ P- ’ \\\\\ I \\ l / l \ / H \/ ‘ (Boys) _ c /3 E—-—-— R l N/5 r0772: Q g E: I /4- J Y—LLL. ’{fi—sr—io a, 37. fl Perl-a Js 6h SUMIAH! This study was made to determine the effect of additional calories in the form of fat, starch. and sugar on the urinary sulfur excretion of six preschool children. The experimental period lasted 102 days with a.preliminary adJustment period of IR days. The entire study included determinations on other minerals. 1. During each diet the total sulfur exeretions varied in proper» tion to the dietary intake-athe children who had the larger intakes had greater sulfur excretions. 2. The per kilogram sulfur excretions also varied in proportion to the dietary intake. 3. Additional calories in the form of fat. starch. and.sugar had no effect on the per kilogram urinary sulfur. n. In general. there was a negative correlation between the per- centage gain in weight and.percentage of dietary sulfur excreted in the urine. The lower percentage gain in weight was accompanied by a higher percentage excretion of urinary sulfur. 5. The girls made greater percentage gains in weight than did the boys on each of the diets. 6. All of the children made the greatest percentage gains on the high fat diet and.the least on the basal diet. 7. The additional calories decreased the N/S ratio in all cases. 8. 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Merle Bray. and.flarie Dye. 1937 Effect of diet on the constancy of the urinary nitrogenous constituents exp creted daily by preschool children. Jr. Nutr.. vol. 13. p. 179. Hanks. J. 3.. Merle Bray. and.flarie Dye. 1938 Influence of diet on the nitrogen balances of preschool children. Jr. Nutr.. vol. 15. p. 125. Sherman. H. C. 1933 Chemistry of Food and Nutrition. Hacmillan Company. New‘York. .. i \D s} .91.. )7 . . z. . 1...... 1 1.1.1: - . ...-.. ......t. . .. ... ‘. eeA~A1v e' . . ‘1‘. r . . ,. L A. . . .. . o ..u ... .e . a . ... . .. - ~1..‘ 1.1. 1 1. I . . a 4 . . , . . . . e . U I . , . n a .ener1II’\ - I. ' I . A A . . ...QNW“. rMI?.\se.e‘Htst x..- ,. 5 ‘9l Junta... . .. . .é‘we. .2 . .. . ......dl: . . 1.1... r. ... . . . k3 WC. . 3.1). . ...... W ..*s.e)nn.4 .MIf 31.3.. 9....93va. .Mmmh..\t.m.¢,.wm.1..flu}1_.4 seekwusifhv . er» 1 3.1!: . h ...r .11.”... - an annexing“... mflflwfi e... .3“..- ... ..Htawr....... . .v . . 1 . dd... 2.: ...“... amen... . .. .. .0 ”as! 1 .1. ....“1‘, . 1 . .‘et. .. I .e’, ”a ,. “feet. 1 V...‘J ., , .. 94- , 2. .1... .. w... . ......v .....r?«.... . t. . ......11..11.......u . - .. . . . . u 1 . x v e . .I . ..., . .. . . .. .... . . A . l a I .. .4? 1. . 7 iv”. .r . .. v v. A . bl .lt. IE.. $y ‘4» . fi “ _ 1 .7 .. ...... .. .....- .... u , _ . . t . e '.L.v 1th chin). «e 1 A. . I I 1 . s . . ,t . 1 . . . u . . t a , . . . . 1 ... . t. .. I . .— s . v .. . . . . 4 e ‘ C . A. 1 V I .. a, I . . . . . . ‘ J .. - . . . .I. I , n 4 . . I . 1 _ . . I a a I I e . . I . . . . .) e P . . . 1 . a . v 1 1 4 I . . a .. . . _ 1 . I . u s I t a s I- I 1 .. . n . .m 1. I u ‘4. 1 e IA '\.tf' 10 “9 .\1| . I: at .1 NICHIGQN STQTE UNIV. LIBRQRIES i W! HIIW HHHI‘JI: II ille l’HHHHl 31293010663478