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THE CALORIE BALANCES OF FEE-SCHOOL CHILDREN AS AFFECTED BY A CONSTANT DIET AND BY AN INCREASE IN THE PROTEIN CONTENT OF THE DIET liltti‘lfi’. *.¢¢O$CI# ##ilifi. tifltfi ‘it t A Theeie Submitted to the Faculty of Michigan Stage College of Agriculture and Applied Science in Partial Fulfillment of the Requirements for the Degree of Master of Science by Jeanne Martin Egprheee Department of Foods and Nutrition Division of Home Economics 1936 T H 553 S ACKNOWLEDGMENT The writer wishes to express her appreciation to Dr. Marie Dye and Dr. Jean Hawks for their helpful sugges- tions and criticisms in directing this experiment and in preparing this report. The writer is also grateful to all other persons whose co-operation made this study possible. 105404. THE FOUR CHILDREN From left to right: J.3., V.B., E.C. ani :.' THE FOUR CHILDREN From left to right: J.H., v.s., 11.0. and 0.3. TABLE OF List of Tables Charts . . . . . . . . . . Introduction. . . . . . . . Review of Literature. . . Procedure . . . . . . . . . Discussion of Results . . Food Intake. . . . . . Study Number 1, Medium Percentage. . . . . . Study Number 1, Medium Basis . . Study Number II, Percentage. . . Study Number 11, B3818 e 0 Period Averages. . . . Discussion . . . . . . Summary . . . . . . . . . . Bibliography. . . . . . . . CONTENTS 0 I O O O O O O O O O O O 0 Protein Diet-—Total O C O O O O O O O O O O O 0 Protein Diet--Per Kilogram O O O O O O O O O O O O O C . High Protein Diet-~Tota1 and 0 C O O O O O O O O O O O C High Protein Diet--Per Kilogram O O O O O O O O O Ul-F'WH 21 38 “3 55 63 72 79 82 55 86 10. 11. 12. 13. 1’4. 15. «.1... LIST OF TABLES Caloric Intakes of Pre-school Children as Reported in the Literature. . . . . Atwater and Inter-Allied Standard . . . A.Rearranged Portion of Bray's Table 3. . . . Rearranged Portions of wang's Tables-~Total Balances. . . . . . . . . . . . . . . . Rearranged Portions of wang's and of Muller's Tables--Balance per Kilogram of Body Weight Height and Weight of Children Compared with 'OOdbul'y . 8 Tablefi. o o o e o e e e e e o 0 Composition of Diets as Calculated From Standard Tables. . . . . . . . . . . . . Quantities of Food for Each Child. Medium Protein Diet 0 O O O O O O O O O O O O 0 Quantities of Food for Each Child. High PrOtein Diet 0 O C O O O O O O O O O O O Fluctuations in the Composition of Various Substances Used in the Study . . . . . . Factors for Changing Cubic Centimeters of Oxygen to Calories . . . . . . . . . . . Variations in Analyzed Calories. . . . . Daily Calorie Balances. Medium Protein Diet 0 o O o O 0 e O 0 e o o e O o o e 0 Daily Calorie Balances, Per Kilogram of Body Weight, on a Medium Protein Diet. . Correlations Between Diet Calories and Caloric Output, Retention and Absorption 16 17 17 22 25 26 27 31¢ 35 39 an 56 5s 16. 17. 18. Daily Calorie Balances, High Protein Diet. . . . . . . 65 Daily Caloric Balances per Kilogram of Body Weight, on a High Protein Diet . . . . . . . . 7} Variations from the Average Retention per Period per Kilogram of Body Weight . . . . . . . . 80 CHARTS 1. variations in Analyzed Calories . . . . . 2. A Comparison of Total Calorie Outputs on a Medium Protein Diet . . . . . . . . 3. A Comparison of Total Calorie Outputs on a High Protein Diet . . . . . . . . h. Variations from the Average Retention Per Period Per Kilogram of Body Weight . no #6 “9 81 THE CALORIE BALANCES or PEI-SCHOOL CHILDREN AS AFFECTED BY A CONSTANT DIET AND BY AN INCREASE IN THE PROTEIN CONTENT or THE DIET INTRODUCTION Since the caloric requirements of pre-school children have interested investigators for many years, children have served as subjects for numerous dietary studies. The early German investigators, as well as the later scientists, ac- tually analyzed the food for calories in a few of these studies, but in the majority of the cases they calculated the values from food composition tables. .The investigators did not analyze the excreta in these dietary studies; thus they did not measure the calories available for the various bodily processes. Nevertheless, these results have in a large degree furnished the basis for determining the caloric ~requirements of pre-school children. Although information concerning the caloric balances of pre-school children is meager, a few studies have been made both in Germany and in this country. The majority of the subjects were greatly under-nourished or very much overweight; therefore these studies did not give results for normal children as compared with the standards of today. -5- Even if there are a few results for normal children, they do not fully determine all the factors which affect the caloric utilization. The main object of the present study was to investigate some of the factors which influence the caloric utilization of pre—school children. The study determined, first, the influence of a constant diet having a fixed caloric intake on the calorie utilization, second, the effect of an increase in the protein content of the diet, and, third, the variation in the reaction of individual children. REVIEW OF THE LITERATURE Early German workers did the first studies of caloric requirements and they prepared standards for children of all ages. The value of these results lies in their historic 'interest rather than in their information. There are several reasons for this. The technique was inaccurate, chiefly because scientific methods of analysis had not yet been per- fected. There was a small variety of implements to work with, and those that were available were not as carefully calibrated as those used today. Thus the chemical technique did not give as correct results as that now available, al- though it was probably excellent for that time. Then, too, - 6 - the data obtained was limited. In many instances the periods were short and the entire experiment extended over an insuf- ficient number of days to show the total range in fluctuations. Often the number of subjects observed was not large enough for comparisons or for definite conclusions to be made. According to the present height-weight-age standards many of these subjects were under-nourished, although some were over- weight, and the majority of them were typical institution children. Table 1 presents in chronological order, along with other data, the data on caloric intakes (as quoted by Holt and Fales (18)) which a number of the early German authors re- ported. In 1881 Uffelmann, one of these workers, reported a study on his own four boys. The eldest and the youngest were slightly under-weight, while the other two were very much below standard. Their caloric intake ranged from 88 calories per kilogram of body weight at one year of age to 68 at five years. Hasse calculated the caloric intake of six girls from 2 to 11 years of age, four being of the pre- school age. Two were underweight children; of these a two- year-old consumed 120 calories per kilo, and a five-year-old 90 calories. On the other hand the other two were three-year- old overweight girls and they used 77 and 81 calories per kilo. Of the early results, Camerer's standards are most widely quoted. He obtained his results from the food con— sumption of his own children and grand-children, practically nonpcwop commaobm manac can whom. ma _ .aa .Hw .sa _. mm omao~ omuow omsom canon omuow m~.oo A as .HN mm.m~ . om .om .m~ oo .- Np m.Hm .om .Hm «.mm .2. jump w an: mafia m m a a a a HM-M~ . mm mm o w mm o m w s me mma maa pm” an mom mom can NHH mma ma ma mm .om am aN ma ma .mm mm ma .NN ma mm om w mos :HH mma mom om ~o ca ma ma .om Na em ma. m~ ow om ”maaa moH mo ma MN cm a Ha..u as. u as. u Hu.na as a sec mm as..m as .x as -g ano.u seams Hue .I'IIII'II 3.30 I'llmlmlololll m cm 090. I sand. 1; on. >¢ on. >4- heum sadauonm amoano: ouwoz «and mucus“: cocoa nowaoomoa sofindooc .aeoauuomaa coach a «How _o>«aos hao> abaao< sense when I mean paomauu soxoog scomuopm nephew nonosmo omncm cumaaouua onspmnoaaq on» an ooanomom a candy no soauaano Hoonomoum no moxepna cascade -8- all of whom were underweight. For boys he allowed 89 calories per kilogram of body weight at one year of age and decreased the value to 75 calories at four years. After the age of A five the per kilogram allowance for both sexes rapidly and steadily decreased until at ages over 15 it was only a little over the basal requirements as determined later by Benedict and Talbot. Baginsky reported data from several children who were convalescent from various diseases. lith but one exception the children were very much undernourished. Thus, the values were for the most part high per kilogram of body weight. Of the six children whom Herbst observed only two boys aged two and four were of pro-school age. The youngest, who was considerably overweight, consumed 90 calories per kilo, and the normal child 87. In 1899, Steffin reported values which were higher than any other scientist had advo- cated. He thought that children needed over 100 calories per kilo for the entire pre-school period. Later, Locke averaged the values found by Uffelmann, Hasse, Steffin, Herbst, Camerer, and other German workers and from these results compiled a'table. He found that the average calorie intake for a child two years old was 9h calories per kilo- gram of body weight, and for a four-year-old, 82 calories. Before the above miscellaneous material, even the standards of Locke, could be used to estimate the food needs ‘of large groups of children it was necessary to)gather it together, organize it, and compile a simple table. Therefore, -.9 - Atwater (l) surveyed the dietary studies made in this and foreign countries and deduced general factors in terms of calories per man per day. The consumption of an average 70 kilogram man doing moderate work was 100 grams of protein and 3000 calories. To this amount he gave the value of 1.0 and to children he assigned fractional portions (Table 2). These he used to calculate the food needs of children. Table 2 Atwater and Inter-Allied Standards Child Age Atwater Inter-Allied 0-2 0. 005 2-6 0.. 0.5 6-10 0.2 0.? Boy 10-12 0. 0.83 Girl ’ 10-1# 0.6 0.83 Boy 12-1“ 0.8 0.83 Boy 1n.1e 0.9 1.00 Girl lu-lé 0.7 0.83 Man 1.0* 1.0' ‘Woman 0.8 0.83 Heavy muscular work ,_ 1.20 ’TEis unit valueETl. was aqua; to 100 grams of protein and 3000 calories. The New York Association for Improving Conditions of the Poor questioned whether the allowances for the food require- ‘ment of children as expressed in terms of man as-a unit were adequate. Therefore Lucy Gillett studied 563 well-nourished children.betssen the ages of two and seventeen (13). She made 223 dietary observations when the subjects ate a freely > - 10 - chosen diet; conducted 131 metabolism experiments and 209 respiration tests. She reported that results of the metap bolism and the respiratory experiments were comparable with those of the dietary observation. She also found that it was necessary to make a distinction in the total caloric requirements for boys and girls after the third year, and, furthermore, if the Atwater Standards were used there should be a range of from 300 to 600 calories above the standard for each age to allow for individual variation. Although her work was thorough she did not have enough material for each age group to make adequate standards of her own. She did, however, report standards (Table 1) determined by averaging her own results with those of previous workers. In 1918, after Gillett had pointed out the errors in the Atwater standard, the Inter—Allied Food Commission re- vised and improved the earlier standard in order to be able to estimate the food needs of the nation during the war. The Inter-Allied Standard left the calorie allowance for men the same, but increased that for women and for all children. (Table 2). Lusk (22) took as his foundation for the calorie require- ments the basal metabolic needs and estimated over and above this the hypothetical number of calories for various degrees of activity. He allowed an increase of 50 per cent for a quiet boy, 100 per cent for an active boy, and 200 per cent for a very active boy. - 11 - Although some of the above standards were suitable for estimating the food needs of large groups, none were ade- quate for determining the calorie intakes of small groups, because there was not enough data for children of various ages. Therefore Holt and Fales (18) obtained the dietary records of 100 children from 1 to 16 years of age who were healthy, well cared for, and normal as to digestion. They kept an exact record of the food intake of each child for four days. The study included five to twelve observations for each child up to the age of eleven, and from these they determined definite standards of total caloric intake for each year of age for both boys and girls. In calculating these standards they considered the calories needed to provide for basal requirements, growth, muscular activity and the loss in the excreta. Since they used the factors u, 9, and h for estimating calories from protein, fat, and carbohydrate,which care for loss in the excreta, they ' allowed the metabolic waste twice. Thus, their allowance for loss in the excreta was higher than it should have been. These various standards give an idea of the number of calories per kilogram of body weight required by children to enable them to grow into normal healthy men and women. Other work was needed, however, to make the standards for pre— school children more dependable. Thus, there have been a number of other studies on children of this age (Table l). E. H. Starling (31) quotes Carl Tiyerstedt who reported - 12 - 47 observations on children ranging in age from four to 1% years. The wide range of calories consumed was the most striking characteristic. In four observations on children in their fifth year he reported a range of from 73 to 11a calories per kilo and in five observations for the seventh year a range of from 71 to 102 (Table l). lhile at the University of Chicago, Goodhue (15) observed 31 superior children from two to six years of age. She computed the caloric content of the cooked food consumed on the basis of the raw foods used in the recipe and employed factors derived from the percentage composition tables made by Atwater and Bryant. A six-year-old boy consumed the maximum number of calories, 2761, and a three—year-old the minimum, or 121% calories. The children had an average total intake of ieun calories or 85 calories and 3 grams of protein for each kilogram of body weight. Messenger (2h) made a dietary study on 13 nursery school children from three to five years of age. She found it necessary to have the mother keep the records of the morning and of the night meals at home and she recorded the food intake of the mid-day meal at the nursery school. With the help of assistants she weighed the food for the noon lunch on a Chatillon balance and recorded the weights. Then each child chose his or her own plate of food. If desired the children selected extra sandwiches and milk. Either the author or an assistant weighed all uneaten food back and de- - 13 _ ducted it from the original amount served. They kept records for an days and divided the time into periods of six days duration and selected for each child that period most nearly average for the entire study and for this single period calculated the calories from Rose's tables. By this method the average total calorie intake for the group ranged from 1153 to l6h0 calories. In their homes, Jones (20) studied four nursery children. She weighed all food consumed and calculated the calorie content from the figures of Peterson and Hoppert for cooked vegetables and reckoned the ash of canned fruit as three- quarters that of the fresh product. The average caloric intake per kilogram ranged from 77 calories for a four-year- old girl to 96.6 calories for a five-year—old boy. The results compared favorably with the standards of various authors excluding those of Lusk which were much higher. At a Chicago orphan asylum, white (37) observed the diets of children of a number of different nationalities. For the complete dietary study 10 healthy children from three to six years of age served as subjects for three weeks excluding all Saturdays and Sundays. She weighed or measured the food and computed the calories from Sherman and Rose's tables. Average figures for the entire study show that the group consumed 170M calories daily which was more than any other author had reported in dietary studies. In a table, lhitesquoted the average intake, 1285 calories, reported by - 1h - Imlay (19) in her thesis. McClelland (23) studied 42 separate daily calorie intakes of two nursery school subjects and paid special attention to the length of time of sampling periods in order to determine a reliable period for dietary observations. She weighed all food as eaten and saved aliquot samples and analyzed them for caloric content. Then she grouped these daily intakes in all possible 3, 4, 5, and 7-day periods and in 2, 3, or u-week intervals. Although the intake varied greatly from day to day it rarely fluctuated in the same direction on two consecutive days. The average intake of the two children was 1309 calories. Her data seemed to indicate that chance selection in a short time study may obscure the wide variations characteristic of the voluntary food intakes of children. Since the mean and the medium were nearly equal, and the number of days above and below each were about the same, the #2 day period constituted an adequate sample. The four and five-day periods were no more reliable than the three-day periods. Consecutive days gave a more uniform and consistent sample of intake than scattered days so that omission of one day in a short time study might make the results fallacious. In the seven-day periods one out of every 12 deviated more than 10 per cent from the average, but none varied more than 15 per cent. In the 14, 21, and 28-day periods all varied within five per cent from the average. Thus a dietary study should extend over an in- _ 15 - terval of not less than two weeks. In 1931 Iinters (38) reported an interesting comparison of the calorie intake of 50 American, 50 Negro and 75 Mexican children. She calculated food intakes and grouped the data according to age, sex and nationality. These groups were fairly evenly divided as to sex but age was unevenly dis- tributed; therefore the figures are only indicative. As the child grew older the caloric intake per unit of weight de- creased. She considered that an intake 10 per cent below Holt and Fale's standard was inadequate. On this basis 52 per cent of the American children, 62 per cent of the Negro and 66 per cent of the Mexican received daily an inadequate number of calories. A large percentage of the children also had the necessary food constituents other than calories in amounts too small to insure an adequate diet. Through a period of years Bray and others (8) accumulated data on the food intake of nursery school children. The sub- jects, 8 boys and 12 girls, varied in age from 33 to 61 months. The authors weighed an exact duplicate of each day's diet and determined the caloric values with the oxy-calorimeter and also by calculations. The difference between the analyzed and calculated values of the composition of the diets varied greatly, from plus 35.h to -27.3 per cent, although the coefficient of correlation between the two was fairly high, 0.8ul. They also found marked variations in analyzed intake, ranging from 879 to 2h22 calories per day (Table 3) with an' - 16 - average of 1399 or 77 calories per kilogram of body weight. Table 1 shows that these average results per kilo are com- parable with those reported by other workers. Table 3 A Rearranged Portion of Bray's Table 3 figetgn Ngmber of}_“ ;T;§Ei3£1g§ on g g 11gggg ot Per Kilogr 1Everage range average -ra:g; ' 0.41 19 1260 879-1691 71 53-100 2-23 26 1440 1081-2422 81 58-128 54. 5 15 1 26 919-2215 77 54-96 2 by; i 23 $3 r s A 60 gotal 1304 18 Although these dietary studies have given much informa- tion concerning the food consumption of children, they have not shown the number of calories of the food ingested which were utilized and those which were excreted. There are only two calorie balance studies, that of Muller (18) and that of lang and her associates (34, 35, and 36) available for com- parison with the data presented later in this experiment; therefore, the author rearranged the results for normal pre- school children and made some additional calculations. Tables 4 and 5 give the results as thus organized. Muller conducted an extensive experiment on 23 boys and 9 girls from two to six years old. He collected com- posite samples of food, feces, and urine for each child during _ 17 - ulIIIMfi. m mm. H.mm m z m m m 0H 0. H museum 0 amuse no a: muom mumm m.H ~.m m.m «.mm m.m Heaaoz H.~Ha m a MM.4 a.mo m.- .H.m m. m.m ~.e~ w.~um._ Henson m.ma .a a _a.m o.o~ ~.m~ ~.m mm.m m.” m.w~ m.e noon: add ~.mH _ a m.a m.mH o.mHH ~.m .M H. :.~uH .a Hashes m.~a m a oo.« w. m m.mm Vmum Wum mnwa o.moH noeoa_ae.m m.ma m a .m.n O . a o \e e e WIFE as as Sigmw 8 ME... we evilwmwmewm .3 twee a melee rmmeom .mmepe. . . -..ao. - .H.- omm. . cameos. ems «on cameo pawns: Room no amawoaau non moseaem nuance u.noHHs: no use o.mceh no anodanom memorandum m canoe m; ”mama ma Hmma m.H mm 3.: on . mam. H ma a we“4 m m as m omHH m.m an ..m mm “N am n ma a .a.m mm a a a oaoa m.a am . on .a an e on a a.a m meow m mawm m.m mm ..m om ..m mus N am a mo.4 w mfifl. m Om.H fiem N .N #NH 3.0H HWH #NQH h .mefl m m a . e u Ho}: 4 a o a 1...”: n a. e u. . ..w "0 .MW “0 NMNH : g m ooaoeeeom aoaaoaoaoa. cease 7:14aammquauaauo .H.e.a a1 .oueaaH new sense Ill< oosmaem Hence scares u.mcsh no ecoaanom domdmnumom : manna -18... a six-day period and analyzed the three samples in a bomb calorimeter. The children consumed an average of 104 calor- ies per kilogram of body weight. The boys used 15 calories per kilo more than the girls who were slightly less under- weight than the boys. Daily, the children excreted an average of 5.9 calories per kilogram of body weight in the feces, and 4.6 in the urine. This was an average loss of from 8 to 14 per cent of the total calorie intake. For the group, the average absorption was 98.1 and the average re- tention 93.5 calories. Tang and her assistants conducted many metabolic ex- periments which included caloric balance studies. These latter they reported in three papers on calories. Although the majority of the subjects were undernourished the controls were normal. Wang et al calculated the calorie intake from tables by Rose and determined the output in the feces and the urine in a bomb calorimeter. For the first study (34) they used subjects over 8 years of age; consequently the results were not comparable with those reported in this paper. In the second study (35), Rang planned two diets which were similar in calorie content, and which were to supply two and four grams of protein per kilogram of body weight respectively. Some children refused to eat all the food served and the analyzed nitrogen content was higher than the calculated; therefore the number of calories in the high protein diet was greater than in the low. The calorie -19.. intakes for the normal pre-school children on the medium diet were 74.7 and 97.2 calories for a four and for a five- year-old girl. They retained 69.4 and 90.6 calories res- pectively. 0n the high protein diet a different four-year- old consumed 76.3 calories and retained 70.0, while the same five-year—old girl consumed 122.4 and retained 115.3 calories. Tables 4 and 5 give the complete data for all the pre—school subjects reported. From this study Tang et a1 drew several general con- clusions. The low protein diet gave a higher percentage utilization of calories than the higher level. In all groups the children had a greater calorie loss in both urine and feces on the high protein diet. The increased calorie value of the urine was due to the higher content of nitrogenous substances such as urea and uric acid, and the increased calorie value of the feces to a larger amount of indigestible connective tissue and to the larger calorie intake on the high protein diet. In a third study (36) Hang and her associates observed twelve children as they gained in weight. Results confirmed the earlier findings, that the absorption of the undernourished children was equal to that of children normal in weight and that it varied directly with the intake. Sherman (29) believed that the standards for the food consumption of children were not suited to short time studies on small groups of children. Practically all of the early - 20 _ standards, including those of Locke, were determined almost entirely from data on under-nourished children. Even the later ones by Gillet and those by Holt and Hales which in- cluded some studies on normal children were greatly influenced by the results from underweight subjects. He thought it dangerous to use the standards of Atwater for estimating the dietary needs of a family including children, because of the wide individual variation in the food requirement of a man according to his occupation and his nationality. Recent dietary studies had contributed some valuable information concerning the wide range from day to day in calorie intake for normal healthy children. Therefore Sherman compiled new standards for the food consumption of normal children. To provide a safe margin he allowed a range of from 90 to 100 calories per kilogram of body weight for the child from one to two years of age, from 80 to 90 for the two to five- year-old, and from 70 to 80 for the six to nine-year-old. Rose quoted these values in her book (28). Although some of the recent dietary studies previously reviewed seem to confirm the authenticity of Sherman's stan- dards there are still many things to be learned about the calorie requirements of children such as the caloric utili- zation, and the factors which influence this utilization. The present balance study should help toward clarifying some of these problems. - 21 - PROCEDURE In this experiment four normal pro-school children, two girls and two boys, served as subjects for two metabolic studies, one on a medium and one on a high protein diet. The experiment lasted for 55 consecutive days. The subjects were similar in age and in physical con- dition. The two girls, J. H. and C. B., were both 36 months old. According to woodbury's tables (39) they were slightly below the average weight for their height and age (Table 6), but within the normal range. It is interesting to note that, although both girls gained equally in weight, the percentage variation from the average decreased for C. B., and increased for J. H., due to the latter's slightly greater increase in height. Since the percentage variation from the theoretical weight varied more for J. H. than for C. 3., her gain was more irregular. Physical examination showed that J. H. had definite signs of infantile rickets which had apparently been cured, and that she also had a somewhat lowered muscle tone. As the experiment continued, however, she improved greatly in muscle tone, in color, and in activity. Since the children both gained in weight throughout the entire interval the diets probably furnished as many calories as normal children receive. Table 6 Height and Weight of Children Compared 'ith Icodbury's Tables no: m :3: :::::::: n I g.. ... .n #01st N 000 00000000 +5 2 + + + + 4 -I- 4 ++ +4-4- g 90 9.1: an o NNN NNNNNNNN [St W. a m dmm wwmmwwww {RB “09‘“ H HHH HHHHHHHH I N wwm wwwwmwww :5 w :r NNN NNNNNNNN . {enact .u to' sound bonoootototohoto H H HHH HHHHHHHH a H mmm mmmmmmmm “IBIOH o F; «NINA mmmmmmnm # ### 0 no ‘33 H©®N#NO O##;EO'60#KJN x: - I: " Ifiunmxoummun. um§=tuxwueuxrx +5 "19A +++++++ ++++4+4+ <32 HHHHNNN Nmmiiidm : IBOtQOI m eeeeeee e H -09 .u .oooxooxoxo \oéxooxé .0on 3 In ‘11 HHHHHHH HHHHHHHH h-an N30 10 mmmmmm mm {P m mooHHoo OOOOOHQO 0 O O O O O O O O O 9 O O m. Imgo' .14 to I-I¢I~I-I~'—I~ NNNNNNNN p HHHHHHH HHHHHHHH mmmmoo ov~r~wwnomm qutsH s . . . .. . .. . c 00000 00000000 # ##### ######## ###~D~OO\O\ ~0# O\\O mmom "3 “0‘1 ‘9‘ NNNHHHH HHOdOHHo-I ‘2 “I134 II II II I I II II II I o +2 mqmmc‘omo‘ "' “ “m" :2 33333:: assume... R‘s ’09III HHHHr-‘l H HH HHHHHH [‘ mmmmmwu mONHNHmH m mmmowxoo. o Ish-no h-wto '60 '[FIIQOV m eOeeeeee , u ##1##:3' »:::::::: o HHHHHHH HHHHHHHH mmmmwmw mmmmmooo QQSIGH a ’ 0 O 0 e e e a a o mmwwmmw wwwmwmmm mmmmmmm mm mm Nd‘##I-IHO\ mnHoH to ,3 not: as. N. . .3“ :1. . . . {20‘ . mmm Ind-Nui- m:- g 'WNM. IIIIIII IIIIIIII O 4’ HHHHNNN F\# ####Lh anaoxqez meeeeeee ass-eeeeee _ s is ####### ##d‘d’d‘d’d’d’ ‘33 0‘11. HHHHHHH HHHHHHHH I. -= ease—aes- awesome 9” tmaovweeeeeee eeeeeeee .u mmmmmmm mmmmmmmm 6 HHHHHHH HHHHHHHH mmmmxoxo to moxcxcxcxoxo 89 B 00.090. Coeeee e “I I H o I-I~I~I-I-I-I- NNIs-NIHNK-to mmmmmmm mmmmmmmm at at 901395 0HI-INM#LI\DI~ e-II-IHNth-to :91 mo. - 23 - The other two subjects were boys; one, V. 3., was 48 months old, the other, i. 0., was 53 months. They were both slightly above the predicted average weight for their height and age, but again within the normal range (Table 6). During the experiment V. B. gained more in height than in weight; consequently the percentage variation from the ’theoretical weight decreased. The children both seemed normal, although without doubt, I. 0. had had infantile rickets. On the fifth day of the medium protein diet I. O. contracted a throat infection and was not able to take his diet for three days. He recovered rapidly, ate all of his food and seemed normal. To rule out all possible effects of his illness all of his data were omitted from the first study. During the high protein study I. O. maintained a constant height and weight. it the time of the experiment all of the subjects were healthy, normal children as revealed by physical and medical examinations. As far as could be determined their diets prior to the experiment were at least typical of the average child. None had suffered from any serious illness. The psychological factors were as nearly normal as possible. Throughout the experiment, the children lived in an apartment in the Home iconomics building under constant supervision. They observed a regular routine in regard to meals, toilet habits, play, hours of sleep and afternoon rest. Every day the children spent several hours in active - 2h - outdoor exercise. The conditions were as near those of a happy home life as could be managed. J. H. finally decided the only thing lacking was a Daddy: In determining an adequate sampling period in dietary studies McClelland (22) found that in lh—day intervals all data varied within five per cent from the average, also that individual periods of three days' duration were as reliable as those for four and five days. For calcium and phosphorus balance studies Porter-Levin (21) reported that it required from 15 to 21 consecutive days to include the whole range in retention variations. Therefore, if these are represen- tative, the present experiment was sufficiently long to indicate general trends as well as short time variations. The experiment consisted of two studies. In the first the children received a medium protein diet which furnished approximately 85 calories and three grams of protein per kilogram of body weight. The first ten days were a preliminary and the next 21 days a collection period, further divided into seven three-day periods. Immediately following this first study the protein content of the diet was increased from three to four grams per kilogram of body weight, while the calories and all other factors were unchanged (Table 7). The second study lasted for an days, which were divided into eight three-day periods. The quantities of food for each child and for both diets are given in tables 8 and 9. In order to have identical food Table 7 Composition of Diets as Calculated from Standard Tables Food Height Calories Protein Calcium. Both Diets Grams Grams Gram Milk 300 552.0 26.n0 0.960 Ralston 20 72.“ 2.22 0.009 Orange juice 200 86.0 0.058 Beef no 62.h 8.52 0.005 Res #0 59.2 5.36 0.02 Peaches 150 70.5 1.05 0.02 Apple sauce 150 235.5 0.30 0.011 Celery 20 2.8 0.22 0.016 String beans 100 2 .7 1.27 -0.026 Tomatoes 100 20.2 2.40 0.010 Potatoes 80 6.# 1.76 0.011 Bread 60 1:7.6 5.82 0.030 Sugar 20 0,0 Total-1 1510.7 55.32 1.137 All Children pCaimine 1 I} of 0.5 0.; Ha iver oi capsule , a, 0.1 _ Total-2 - .5 .1 led. Prot. Diet Butter , 20 151,6 0,29 0,003 0,003 _ Total-3 20 153. 0.20 0.003 0.00} High Prot.Diet Butter 10 16.9 0.10 0.002 0.002 Egg white 80 3.8 9.:fi 0.012 0.011 0.0010 Gelatin 10 3,1 1 TOtfll-u e e e 6. 613 Be 0610 Med. Prot. Diet , Totals 1 2 a 3 1669.0 55.e2 1.190 1.27u b.0111 . High Prot. Diet _ : Totals 1 2 & u 1669.6 7h.50 1.201 1.26h 0.0121 Quantities of Foods for Each Child, _ 26 - Table 8 Medium Protein Diet Apple sauce Celery ' String beans! Tomatoes Potatoes Butter Sugar Bread later Carmine Halivsr oil 1 capsul 1 capsul v.3. 3.0. r s r s 37 0 3368 18 20 - 180 200 6 #0 6 1&0 135 150 135 150 18 20 90 100 90 100 72 80 18 20 18 20 54 60 300 300 1/3 of 1/3 of 0.5 sin 0.5 8m 1 capsul 1 capsul - 27 - Table 9 Quantities of Food for Each Child High Protein Diet Beef 28 32 36 #0 Egg 28 32 36 #0 Peaches 105 120 135 150 Apple sauce 10, 120 135 150 Celery 1 16 18 20 String beanei 70 80 90 100 Tomatoes 7O 80 90 100 Potatoes 56 an 72 80 Butter 7 8 9 10 Sugar 1“ 16 18 20 Bread #2 #8 5M 60 Egg white 56 6k 72 80 Gelatin 7 8 9 10 Water 200 200 300 300 Carmine 0.5 in 0.5 in 0.5 in 0.5 in 3 days 3 days 3 days 3 days Haliver oil 1 capsul 1 capsul l capsul 1 capsul E. C. received the diet as calculated. V. B. received 0.9, C. B. 0.8, and J. H. 0.7 of the amount given I. 0. - 28 - intakes, the children received the same amount of each food per kilogram of body weight and the ratio between the quan— tity of each food given was the same as the ratio between the weights of the various children. For examp1e3the weight of E. 0., the largest child, was taken as unity and given a value of 1.0. Since V. B. weighed approximately 0.9 as much as E. 0., C. B. 0.8 as much and J. H. 0.7 as much they were given these proportions of the diet calculated for E. C. By this method of figuring it was possible to give within experimental error 85 calories for each child per kilogram of body weight. To show the effects of a change in protein content, the two diets were as nearly as possible identical. The majority of the foods, in both kind and amount, were the same on the two studies. The only difference was that in the high pro- tein diet, gelatin and egg white increased the protein con- tent and the omission of a part of the butter kept the calories at the same level. These changes also produced slight variations other than those desired. The increase in protein changed the calorie to nitrogen ratio, and the de- creasein fat, the carbohydrate to fat ratio. These, how- ever, were not radically altered. The gelatin and egg white increased the percentage of animal protein. Butter supplied more fat soluble vitamins than the egg white, but the chil- dren were probably receiving an adequate supply from other sources. The residue was the same because the cereals, _.29 - fruits and vegetables remained identical for both diets. Minerals were probably slightly increased by the protein products which replaced part of the butter, and made only slight alterations in the acid-base relationships. In order to insure uniform quality, sufficient amounts of foods to last during the entire experiment were pur— chased whenever possible and in all cases enough was obtained for a three-day period. Each type of food was thoroughly mixed to secure a homogeneous sample and then from this the individual portions for the children for each of the three days in the period and for the two samples for analysis were weighed on the first day of the period. The foods were prepared by the following methods: all water was distilled, milk was thoroughly mixed, the orange juice was squeezed on an electric squeezer, sieved and stirred, visible fat was removed from the beef which was then ground and mixed, Irish potatoes were boiled and put through a ricer, the eggs were broken together and beaten, and the celery was cut in small cubes and mixed. A sufficient number of the canned products for one period were opened and the contents thoroughly mixed. Cerber's pureed string beans and tomatoes were used as pur- chased. Peaches and apple sauce were pureed. The whole wheat bread was cut in such a way that the various samples contained approximately the same amount of crust and soft inside part. The gelatin, ralston, butter and sugar were weighed as purchased without any special preparation, except - 3o - that the same pound of butter was used for all samples in one experiment. Ten grams of the total calculated amoung of sugar was in the powdered state. This was used in making a candy to contain the 0.5 grams of carmine which was given at the beginning of each period to mark the feces. One capsule of Park Davis' Haliver Oil, 250 D, was given to each child daily. An attempt was made to reduce food losses to a minimum. Each food was weighed individually into marked dishes in which it would later be cooked and served, or, in the case of dry foods, onto the waxed paper in which it was to be wrapped. Weighed portions were never transferred from one dish to another when there was the least tendency for par- ticles to adhere to the sides of the container. Samples were well covered with oiled paper and stored in a refrigera- tor until used. ’ The children cooperated excellently by eating all the food served. They did not object to the monotonous diet, probably because the food combinations were varied as much as possible withdut complicating meal preparation. After each meal the dishes were scraped with a spoon, wiped out with bread, and finally rinsed with distilled water which was added to another food that the children had to eat on that day. Duplicate samples of food in amounts given to J. H. were saved for analysis and treated as described later in - 31 - this section. Since all children received equal amounts, Haliver Oil and carmine were not added to these samples. Duplicate samples of food, each equivalent to the amount received by J. H. (Tables 8 and 9), were weighed into enamel- ware pans. They were partially dried on a steam bath, and then placed in an oven at 600 C. until they had reached a constant weight. The dried material was ground several times in a food chopper, then passed through a fine meshed sieve. If small hard pieces remained they were crushed in a mortar, then sieved. The final product was sifted several times through a larger mesh sieve to insure thorough mixing and then stored in st0ppered bottles. All excreta were carefully collected and preserved for analysis. Carmine was used to mark the feces into three- day periods. The sample for each period was saved in an enamel—wars pan and treated by the same method as the food. Urine was collected in Zu-hour specimens. After its' volume and specific gravity were taken it was made up to a definite volume each day with redistilled water. Triplicate samples, each containing 15 cubic centimeters of the diluted urine were pipetted into shallow evaporating dishes which contained about 0.1 gram of salicylic acid (which is a preservative asnll as an aid to combustion). The samples were dried overnight before an electric fan and urine for the second and third days were added to the same dishes. After the entire sample was thoroughly dry, the dishes were - 32 - stored in a covered box until analyzed. The oxyacalorimeter described by Benedict (5, 6) was used in burning the samples. Before starting the analysis the machine was tested and standardized. In order to de- termine if there were leaks in the circuit a weight was placed on the partially filled spirometer bell and the blower started. It was assumed that the circuit was air- tight if readings taken at several minute intervals were constant when there was no temperature change. The carbon-dioxide absorption power of the soda lime was tested by blowing the breath through the soda lime into a concentrated solution of barium—hydroxide. The absence of a milky precipitate indicated that the carbon-dioxide was all absorbed. The area of the spirometer bell used was 20.61 square centimeters. This factor was employed in converting the millimeters of oxygen as read from the scale into cubic centimeters. A definite procedure was used in running the oxy—calori- meter. A small amount of the substance to be burned was weighed by difference into a nickle crucible. It was then placed in the machine, number 30 iron wire was bent into shape so that it just dipped below the surface of the material, and a small amount of powdered pumice was placed at points of contact to prevent scattering and to insure ignition. After starting the blower and before the spirometer bell was _ 33 - finally filled the circuit was flushed out four times with .small amounts of oxygen. This was done to insure an atmos- phere of pure oxygen undiluted with outside air. To provide an even temperature the motor was run for 10 minutes. Then the barometer pressure, the spirometer reading and the temperature of the circuit were recorded; after this the sample was burned completely. Before final readings were taken the motor was run until equilibrium had again been established. This cooling process was sometimes hastened by wrapping clothes wrung from ice water tightly around the hot pipes. They were always removed at least five minutes before taking the final readings. This same technique was used for all substances burned for this study. Before any determinations for the calorie balance were made, the efficiency of the machine was tested with pure sucrose from the Bureau of Standards, washington, D. C. From 1.5 to 2.0 grams of sugar were weighed into a nickle crucible and burned as described above. The results from the combustion of 16 samples of sucrose showed a range from 769.26 to 790.79 with an average of 782.15 cubic centimeters of oxygen per gram (Table 10). This average represented 99.57 per cent of the theoretical yield; therefore there was an average error of 0.43 per cent for each determination. A correction for this error was made on every sample. - 3n - 7 Table 10 Fluctuations in the Composition of various Substances Used in the Study m .15 A .H -‘sll E; 'E 51% a) ‘<533§: 118‘: 3 .3 e as“ a an. ass» as +3 .. It: E400 H +3 .34 0 go- Ho one Hm :3 a: :3 E3 2;°'“ .3 guzrose 16 782.15 769.26- 790.7 765.5 99.57 3.9903 s a naphthol 5 1788.65 l777.57-1811.8 787.5 100.06 Salicylic . acid 12 1129.41 ‘1110.11-1163.9 135.6 99.45 5.280 Carmine 3 819.1 cor. 811.99- 822.8 3.952 Haliver ‘ 817.5 ea. oil # corrected . 761.23- 865.7 . 3.83h3 A portion of the dried food was heated overnight at 60°C to drive off any moisture that might have been adsorbed and triplicate samples of 2.0 to 2.5 grams were weighed by dif- ference into nickle crucibles and then burned according to the method described. In order to convert cubic centimeters of oxygen into calories the factor “.825 (Table 11) calories per liter of oxygen, which Benedict gave for a mixed food was used. - 35 - Table 11 Factors for Changing Cubic Centimeters of Oxygen to Calories Substance Burned Cal. per Liter Oxygen Food 9.825 Feces 9.970 Urine n.680 Sucrose 5.080 Beta-naphthol Salicylic acid n.650 Carmine 4.825 Haliver 011 n.690 Since Haliver Oil was not added to the food sample and was given in equal amounts to all the children it was analyzed separately and corrections made. Four determinations gave an average of 3.8 calories per capsule (Table 10) when the factor “.69, which Benedict reported for fat was used in the calculations. After reheating a part of the dried feces sample, triplicate samples containing about 1.5 grams each were weighed for analysis. According to the factor determined by Benedict one liter of oxygen will oxidize H.97 calories from faces. This factor was used in this experiment. 'Carmine, which was used to mark the feces, was also analyzed for calories. Table 10 shows that triplicate samples checked closely, giving an average of 3.95 calories per gram. It was assumed that one liter of oxygen would give off 4.825 - 36 _ calories in burning carmine, the same amount as would be given off in the combustion of a mixed food sample. The dried urine was transferred from the evaporating dishes to nickel crucibles with redistilled water and ignited asbestos. Ivory tipped forceps and a glass medicine dropper aided in the process. The samples were again dried before an electric fan for at least 2% hours, and then placed for 12 to in hours in an evacuated discicator over concentrated sulphuric acid. A weighed sample of Beta naphthol which contained about 0.2 grams, and a small amount of powdered pumice were added to each sample. A sharp instrument was used to loosen the asbestos so that the above mixture could sift down. After the sample was burned correction was made for the oxygen consumed in oxydizing the salicylic acid and the Beta-naphthol (Table 10). The factor given by Benedict, “.68 calories per liter of oxygen for urine, was used. Before the salicylic acid and the Beta-naphthol were standardized, they were dried over concentrated sulphuric acid in a descicatcr. When the samples for these two sub— stances were burned alone it was found that they scattered badly and did not burn completely; consequently ignited asbestos and pumice stone, in relatively large amounts, were carefully mixed with each sample before it was ignited. In this way oxidation was slowed up so that combustion could be complete. This difficulty was not present during ignition of urine samples because the substances were mixed in much - 37 - smaller amounts, about 0.1 gram of salicylic acid and 0.2 gram Beta naphthal for each sample. The average of 12 de- terminations on salicylic acid was 1129.91 cubic centimeters of oxygen per gram (Table 10) or 99.95 per cent of the theoretical value., Five tests on Beta-naphthal gave an average of 1788.65 cubic centimeters per gram, or 100.06 per cent of the theoretical (Table 10). Because such small amounts of salicylic acid and Beta-naphthal were used in the urine analysis and since the determined values were so near the theoretical yield, the theoretical values were used in correcting for both the salicylic acid and the Beta-naphp thal. \_H -3, DISCUSSION OF RESULTS FOOD INTAKE In this study of the calorie balances of 4 normal pre-school children the estimated calorie value of each diet, both on the medium and on the high protein level was the same. The calculated values for the calorie intakes on the two studies varied only 1 calorie (Table 7) being 1166 and 1167 calories for J. H . respectively. As formerly stated the two diets contained the same quantity of each food except those necessary to change the protein content. Because the medium and the high protein diets were similar they are discussed together in this section. Since the calculated calories for each period on the medium and on the high protein diet were the same, it was thought that the analyzed diet would contain the same number of calories each period, but the analysis showed fluctuat- ions. There were even variations in the duplicate samples of food collected for each period. Chart 1 shows the fluctuations both between the duplicate samples and from period to period. There was a significant difference between the duplicate samples for some of the periods. The least difference between the calorie content of the samples for any one period was 2 calories in period 4 of the high protein diet, and the greatest was 42 calories for period 5 of the medium protein diet. The above values represent a percentage difference of from 0.2 to 3.6 per cent between the duplicate Table 12 Variations in Analyzed Calories '8 a) «£8 0 of“ 3"“? m rim c>N o I} 0:6 g:h0) #1 0 new #> tnmsm cibam -p a 01m 49 93 m +’ Um mHhD G a awn c mam mam ,q C)43® $3 046 c: met: me) “(6:8 0 (DECSCC a) (0601—4640360 0 $180360 (1) H H (D .40 NH 03H (3 noa+1 o a p.6mm (3 Beam :3 ng® 0 cam-1 >~H cH ‘9 “”4550 ®¢SE4$ 5.1 (3:438 H ‘ng H 3% Q3 38: S 3&3: $3 ngrwww o H H> m HEW m (th c3m cIO D4 e1 m in o cue-m a. CI mt react in just the same way, the average total calorie output increased with the calarie intake and therefore with the size of the child. In order of increasing size the subjects excreted 98.5 calories for J. H., 105.7 for C. H., and 109.0 for V. B. Differences between these average Values were less than the period variations for individuals. Nevertheless the per- centage of the intake excreted decreased from 8.5 per cent for J. H. to 7.3 per cent for V. B., therefore the total output did not increase in the same proportion as the intake. The older children may have needed more calories in pro- portion to size than the younger ones. Bray and her associates -51... (8) found that the older nurseryschool children used the largest number of calories per kilogram of body weight: As compared with the study of Wang and her co—workers, (Table 4). the above total outputs were higher than one value and lower than another. One of their 4 year old subjects excreted 87 calories or 731 per cent of the intake, the other, 112 calories or 6.7 per cent of the intake. Both of these percentages were lower than any average value for the present study. Output in feces: More than half of the total output was excreted in the feces. Generally the fecal output varied from period to period in the same proportion as the total output. The differences between the high and the low values for each child, however, were not the same as those for total output. This fluctuation was least 61.7 to 66.6 calories) for C. B., and most (57.1 to 66.8 calories) for J. H. The average calorie output in the faces for the dif— ferent children varied much less than the individual fluctuations discussed above. There was a difference of only 2 calories between the extreme outputs instead of 9.7 calories for J. H., the greatest difference between individual fecal outputs. J. H. had the lowest average output in the feces, 62.7 calories or 5.4 per cent of the intake; C. B. -52- had the highest, 64.5 calories, or 4.8 per cent of the intake; and V. B. had 63. 8 calories, or 4.3 per cent. This indicates that the children did not excrete the same proportion of calories in the feces and, as mentioned before, may have been due to the character of the stools. Output in urine: Roughly estimated there were about two— thirds as many calories excreted in the urine as there were in the feces. Furthermore, individual variations were less pronounced than in either the feces or the total output. Fluctuations in the urinary output were greatest for V. B., (40.3 to 48.4 calories) and least for J. H. (34.1 to 37.2 calories) The periods of high and of low value did not seem to come in any difinite order, although there was a slight tendency for periods of higher intake to also have a higher urinary output in calories. The average percentage of the calorie intake excreted in the urine was practically identical for the three child- ren; 3.1 per cent for J. H. and for C. B., and 3.0 per cent for V. B. On the other hand the average values for calories excreted in the urine increased in exact proportion to the intake. This increase was from 35.9 for J. H. to 45.2 calories for V. B. Thus the number of calories excreted in the urine was in direct proportion to the intake while those excreted in the faces were practically the same for each child. _ 53 - Total absorption: Total calorie absorption measures all the calories ingested in the food which pass through the walls of the alimentary tract and excludes those excreted in the feces. Since the fecal output was so small the variations did not effect the calories absorbed, and thus they seemed directly preportional to the intake. The ab- sorption would also vary inversely with the fecal output. The widest rangeigbsorption values was from 1415 to 1476 calories for V. B. This gave a difference of only 61 calories between the highest and the lowest values. J. H. and C. B. had narrower ranges and therefore smaller differences between the two extreme values, 52 calories in both cases. The variation between the low and the high number of calories absorbed by each child compared favor- ably with similar differences between extremes in intake. V. B. had the same difference in both intake and absorp- tion, while for J. H. it was 4 calories less in intake than in absorption, and for C. B. it was 3 calories more. There was an increase in average total absorption'for the subjects which corresponded to the higher calorie content of the diet and to the size of the child. Average figures show that J. H. absorbed 1106 calories, C. B., 1271, and V. B. 1435. This increase in absorption with the higher intakes was the same as the increase in the size of the subjects, therefore the average percentage of the total - 54 - intake absorbed was about the same for C. B. and V. B., 95.2and 95.7 per cent respectively, and only slightly less, 94.7 per cent, for J. H. Wang and her associates reported (Table 4) similar values (95 and 96 per cent) for two pre- school children. Total retention: Retention values give the number of calories used in the body and vary inversely with the total putput. Again, since the output was such a small part of the intake, the slight variations did not influence the retention greatly. In retention, as in absorption and intake, V. B. had the greatest variation between the low and high value with a dif- ference of 65 calories. This was about 10 calories difference more than that for the other two children. From period to period the percentage of the intake re- tained was nearly constant for each child. This percentage retention was inversely proportional to the percentage of‘ total excretion and the differences were directly related to the fecal variation, because the percentage of the diet ex- creted in the urine remained constant. The average retention varied directly with the intake, and indirectly with the total output.‘ It was least, 1070 calories, for J. H. and most, 1390, for V. B. On the other hand, average percentages of the intake retained varied only about one per cent; J. H. had the smallest value (91.6 per cent) and V. B. the largest (92.7 per cent). These results are comparable with those which Wang and her associates reported for two pre-school children and diets sim- - 55 _ ilar in protein content, but varying widely in calorie content. The children whom they reported upon retained 1146 and 1549 calories, or, in both cases, 93 per cent of the intake. Thus retention probably names directly with the intake when the calorie consumption fluctuates, pro- viding other constituents of the diet remain unchanged.. Study Number 1 Medium Protein Diet-Per Kilogram Basis Intake: Since the daily calorie balances expressed in terms of calories per kilogram of body weight present the data an a comman basis, the results are more comparable for the various children. Table 14 gives the data on this basis. There were considerable differences between the intakes of the children due to several causes. The children,changed slightly in weight during the experiment. Thus the factors used in calculating the intakes were not always in the same proportion. These factors were not exactly proportional to the weights of the different children even at the begin- ning of the experiment as they were carried out to only one decimal. Other slight variations were due to the inability to weigh fractions of a gram of food. Therefore the child- ren did not receive exactly the same number of calories per kilogram of body weight. There was also an individual variation from period to period. The greatestdifference between the lowest and _.55 - Table 14 Daily Calorie Balances per Kilograms of Body Weight on a Medium Protein Diet ggiid Weight kgs. . J.H. 1 13.60 88.04 6.74 4.20 2.55 83.84 81.29 2 13.04 84 00 7.07 4.57 2.50 80.03 77.53 R 13.64 85.50 7.2 4.60» 2.04 80.90 78. 13.64 786.77 7.57 4.90 2.66 81.87 79. 21 2 3'21? 2'? 73338 3.7.3 33"” 7M3 : E: 7 7:15 11:39 a: :3 7 9 7.2 4. 2. 94.01 7.19 4.42 2. 77 89. 59 86. 82 2 90.00 7.12 4.43 2.63 86.11 82:38 .48 a 91.57 7.12 ”-4 2-75 87 13 14 06 92.23 7.5 4.47 3.07 87. 77 84.70 2 14 06 90.42 7.54 4.5 3.00 85.8 82.8 14 02 91.52 7.07 4.32 2.76 87. 20 84. . 7.03 4.22 2. w '” 7.2' 4:4. "2._ 1' 16 97 90 02 6 0 3 68 2.37 86.97 84.60 E 17.05 80 9 6 3 71 2.73 83.25 80.51 5 17 12 87.08 6 73 98 2.75 8 .10 80. 35 0 , 2.83 7 2.53 2. - 57 - the highest intake for any subject was 4.20 calories for C. B. The smallest individual intake was 84.08 calories for J. H. and the largest, 94.01 for C. B., thus the fluc- tuation was about 10 calories. All of the lowest values for calorie intake were in period 7 and the highest in period 1. The average intakes for the children varied in the same proportion as the individual differences. J. H. con- sumed 85.70 calories per kilo, C. B., 91.45, and V. B., 87.88 calories, thus there was a difference of only five calories between the lowest and the highest average figure. This was probably within experimental error. The above values were similar to those reported in food intake studies (Table 1). Of the more recent authors, Goodhue reported an average intake of 85 calories per kilo and Bray et a1 82, while J nes found a higher value, 91.7 calories. Sherman allowed a range in intake from 80 to 90 calories per kilo for children from tflv to three years old, and from 70 to 80 for those from.three to four years. In calorie balance, Wang and her associates found that average figures gave intakes of 74.7 calories per kilo for a four year old girl and 97.2 calories for a five year old. These two subjects were on a diet similar in protein content to the present study. Muller reported an average intake of 104 calories per kilo. Since the subjects in the present study gained in weight when given a diet similar in calorie content to these reported in the literature they probably received an average diet. Total output: Even on this weight basis, there were period to period variations in the calorie output. The greatest was from 6.74 to 7.63 for J. H. and the least from 7.03 to 7.54 calories for C. B. These variations did not hear any definite relationship to fluctuations in intake. Table 15 clearly shows this lackof interdependence as the correla- tion for the two was only 0.04. Table 15 Correlation Between Diet Calories and Caloric Output, Retention and Absorption Intake 7 Intake Diet Period and and Output bsor tion Medium Protein all 0.04 0.94 High Protein all 0.48 0.95 0.96 Med. 8 High Prot. all 0.23 0.97 0.95 - 59 - In spite of the period variations the average total calorie outputs per kilogram of body weight were remarkably similar for the children. For J. H. and C. B. the values were nearly identical (7.22 and 7.24 calories) and for V. B. the figures were slightly less (6.39 calories). These variations were so small that they might have been due to experimental error. The results were slightly higher than Wang and her associates reported (5.3 and 5.0 calories) for two four year old girls, one of whom was from the underweight group (Table 5). The values were also higher than a normal 5 year old girl excreted (6.6 calories per kilo). Muller found that the average daily loss of calories was 10.5 per kilogram of body weight. This was for above any individual figure for the children in this study. Output in feces: So few calories were excreted in the feces that the differences were very small and probably due to experimental error. The greatest difference between the highest and the lowest output in the feces was only 0.7. calories for J. H. and the smallest 0.32 for C. B. These variations were slightly less than those for total output for the same children. Period to period fluctuations did not vary with the intake. The variation in the average number of calories excreted - so - in the feces of the various children was also relatively small. Average figures show that J. H. exereted 4.59, C. B. 4.41, and V. B. 3.74 calories per kilo. In each case this was about two-thirds of the total number of calories excreted. These values were comparable with those Wang and her associates reported for two normal children. One four year old girl excreted, in her feces, an average of 3.2 calories per kilo when her intake was 74.7 calories, and a five year old 4.7 calories when her intake was 97.2. Another five year old whose protein intake was not given, but who consumed 106 calories per kilo excreted much more, 7.9 calories per kilo. Muller also reported a higher average figure, 5.9 calories per kilogram of body weight when his subjects consumed an average of 104 calories per kilo. Output in urine: A smaller number of calories per kilo were excreted in the urine than in the feces. Furthermore, individual variations in the calorie output in the urine were less from period to period than those in the feces or the total output (Table 14). Fluctuation was greatest for V. B., (from 2.37 to 2.83 calories) and least for J. H. (from 2.50 to 2.73 calories). Subject J. H. also had the least variation-in intake but the greatest in total output and in calories excreted in the feces. - 61 - The variationsin the average calorie excretion in the urine per kilogram of body weight were so slight that they might easily have been due to technical errors. The average excretion was greatest, 2.83 calories, for G. B. and least, 2. 63 calories for J. H.; values which are again preportional to the total intake. Since the number of calories excreted in the urine was so nearly uniform for all of the subjects the low total output for V. B. was thus due entirely to his lower excretion of calories in his feces. Wang and her associates reported two values for urinary excretion, 1.9 and 2.l.calories per kilo, which were slightly less than any for the present study. These lower urinary outputs were not entirely due to a lower intake in relation to age, as the child who excreted 1.9 calories consumed 97.2 calories per kilo, or slightly more than any of the children on the present study. 0n the other hand, Muller found that his subjects excreted much more in the urine than any of these subjects, 4.6 calories per kilo when the diet furnished 104 calories per kilogram of body weight Absorption: The number of calories excreted in the feces was so small in comparison with the number consumed that variations in absorption did not show, if they were present. Since this was true there was a very high correlation of - 62 _ 0.94 between the intake and the absorption per kilogram of body weight (Table 15). In every case the absorption varied inversely in proportion to the fecal output. For each of the subjects in the present study there was a diff-' erence of only about 4.0 calories between the highest and the lowest absorption value. This was about as great as the fecal output and greater than the urinary output, but about the same as the difference in intake from period to period. For every kilogram of body weight J. H. absorbed an average of 81.11 calories, 0. B. 87.04 and V. B., 84.14 or about 92, 92 and 93 per cent of the intake per kilo respectively. On a diet somewhat similar to this, Wang and her associates subjects absorbed 71.5 and 92.5 calories which were 96 and 95 per cent of the per kilo intake. Muller reported data, which, when further calculated, gave an average absorption value of about 98 calories per kilo or 94 per cent of the intake. This variation in the number of calories absorbed may have been due to the higher levels of intake. Retention: Since the intake was relatively constant and the total output was small the correlation (Table 15) between the intake and retention was high (0.94). This contrasts well with the correlation between the intake and the total output which was very low because of unavoidable errors and the small figures for output. Individual varia- tions in retention per kilogram of body weight were very small and practically the same for all of the children. - 63 - Flucluations were greatest ( 76.92 to 81.29 calories) for J. H. and least(82.77 to 86.82 calories) for C. B. This variation was in inverse proportion to the output. Average retenton values were quite uniform for the various children. They were lowest for J. H., 78.48 calories per kilo and highest for c. 9., 54.21 calories, Thus they were proportional to the intake. Intake apparently governs the utilization of calories because the output values were such a small proportion of the intake. The large proportion of error in fecal output may make the total output seem more irregular than it really is and may hide a correlation with intake. Wang and her associates reported results that would lead to the same general conclusions. They also found that for normal children the total output was slightly higher for the child receiving the high intake than for the one on the lower calorie'diet. Thus a four year old child retained 69.4 calories when on a relatively low calorie intake, and a five year old 90.6 when on a higher intake level. There were not, however, a large enough number of subjects on one level of calorie intake for comparisons. Study Number 11 High Protein Diet-Total and Percentage Intake:Since the preliminary period for E. C. was sufficiently long he served as a subject for the study in which the children received a high protein diet. This gave four - 64 _ children for comparison while ther had been only three on the first study. Individual variations in intake from period to period were greater on the high protein diet than they had been on the medium protein level (Table 16). The maximum fluct- uation in calorie intake was 94 calories for E. C. and was proportionally less, in relation to the size, for the other children. As formenly pointed out there was very close agreement between the averages of the analyzed calories in thenedium and in the high protein diets. On the second study J. H. consumed an average of 1168 calories, 0. B. 1334, V. B., 1500 and E. 0., 1667. These compare favorably with those reported by other authors. Since the variation was small between the two diets, any differences in utilization would be due to the change in the protein content. Total output: The proportion of the intake excreted on the high protin diet was low, from 8.7 to 9.5 per cent. This was, however, about 1 per cent greater than that on the medium protein diet. As in the previous study the output for each child varied from period to period. Three of the children had rather large individual fluctuations in calorie output, the fourth, J. H., had a relatively uniform excretion of calories with a difference of only 7.8 calories between the EI-.IH.H. «do 53 Tum meg 3:. Mime 0.3 a.m a.m: 33 m m.om mmoa o. m m as m.: a.m a.m m.oe m.m m.mHH News N 0.0m seem 0.3m n om 0.: H.e m.m H.ne m.m N. 0H mmfla m.om Hooa m.:m HHHH m.m o.o m.m w.oo .m m.oHH NNHH m m.om mmoa 0.:m mmofl m. a.m: a.m a.me a.m e.eoH HeHH H & Hue & Hue & Hue * ado * Hue duo .m.h “HHHWUHHHulIIik s «new I:m anI nodpnoaom noapqnonnd. wean an pumowwnoa .IIHIL oxmpan uumammp «can edoeoum swam nooamHmm oaHOHmo madam ma canes Table 16 High Protein Diet Daily Caloric Balances Caloric Output - i n I $21138 Intake Feces Urigg__ Absorption Retent o J.H. cal cal % cal 1% cal % cal ; 081 % 1 1161 106.4 9.2 62.6 5.4 43.8 3.8 1098 - 1055 90.9 2 1172 110.8 9.2 60.8 5.2 0.0 .3 1111 -3 1061 90.5 3 1156 10 .2 9. 63.1 5.5 4.0 1093 .6 1047 90.0 1202 11 .2 9.5 60.5 5.0 5% 4.5 1142 .0 1088 90.9 5 114 112.1 9.8 63.0 5.24 4.3 1082 .5 1033 90.2 6 118 111.7 9.4 6 .4 5. 4.0 1119 .2 1072 90.5 7 1136 110.8 9.8 64.4 5.7 40. 4 4.1 1072 - 1020 90.3 8 1186 108.1 9.1 64.4 5.4 43. 8 3.7 1122 .b 1078 90.9 Average 1168 110.4 9.5 62.9 5.4 47.6 4.1 1105 .6 1058 90.6 0.3. ' 1 1 26 122, 9.2 0. . 52.0 .9 1250 .7 1204 90.8 2 1334 112.8 8.4 54.6 2.1 58.0 3.3 1284 .9 1226 91.6 a 1321 123.8 9.4 68.9 5.2 55.0 4.2 1252 .8 1197 90.6 1373 129.8 9.5 74.8 5.4 55.0 4.0 1299 .6 1244 90.6 2 1308 126.1 9.6 69.6 5.3 56.5 4.3 1238 .b 1182 90.4 1352 124.6 9.2 67.9 3.0 56.7 4.2 1284 .0 1228 90.8 7 1298 117.4 9.0 62.2 .8 53.2 4.3 1236 .2 1181 91.0 8 1355 116.1 8.6 61.4 4.5 5 .7 4.0» 1294 .5 1239 91.4 ‘g 7=:verage 1334 121.6 9.1 66.2 5.0 55.4 4.2 1268 .0 1213 90.9- v. s. . . 1 1491 124.7 8.4 63.3 4.2 61. 4 4.1 1428 95.8 1367 91.7 21206 131.0 8.7 69. 4.6 61.6 4.1 1439 95.4 1375 91.3 4 1 85 1 2.3 8.9 68.7 4.6 63h 6 4.3 1417 92.4 1 53 91.1 1545 1 3.2 9.3 79.2 5.16 4.1 1465 9 .8 1 01 90.7 2 1 71 129.3 8.8 70.1 .8 59.1 4.0 1401 95.2 1342 91.2 1 21 128.2 8.4 67.4 4.4 60. 4.0 ' 1453 95.5 1393 91.6 7 1 60 128.0 8.8 64.6 4.4 63.4 4.3 1395 95.5 1332 91.2 8 1524 123.2 8.1 60.8 4.0 62.5 4.1 1493 96.0 1 01 91.9 Average 1500 130.0 8.7 67.9 4.5 62.1 4.1 1432 .5 1371 91.3 E.0. ' 1 1657 147. 8.9 86.9 5.2 60.2 .6 1570 .7 1 10 91.1 2 1673 159 9.5 90.7 5.4 68.7 4.1 1582 .6 1§13 90.4 3 1650 162. 5 9.8 91.3 5.5 71.2 4.3 1559 . 1 88 90.2 1716 161.1 9.4 95-5 5.6 65.9 3.8 1620 1 55 90.0 2 ' 1934 161. 9 9.9 101.0 9.2 90.9 3.7 1533 1 72 90.1 1689 164-.6: 9.7 98.3 5.8 66.3 3.9 1591 1,25 190,3 7 1622 159. 5 9.8 94-2 5.8 65.3 .0 1527 1262 90.1 8 1693 155.9 9.2 90.7 5.4. 65.2 3.9 T1602 1537 90.8 Average 1667 i 159.0 9.5 93.6 5.6 65.4 3.9 1573 1508 90.51 -99.. - 66 - smallest and the largest values. There was a low correlation between intake and output as shown by chart 3. The curve for E. C. was quite dif— ferent from those of the other children. It indicated a higher output, was more smooth, and reached its highest point in period 6. For the other three subjects the great- est output come in that period in which the intake was greatest, or period 4. At no time did-the periods of least output and lowest intake coincide, although the latter was seldom the same on any two periods. The output curve for J. H. was nearest to being a straight line, and, therefore, more like the curves for the first study. The relation between the intake and the output was also shown by the percentage of the intake excreted, which was more constant for J. H. (9.1 to 9.8 per cent) than for any of the other children. Fluctuations were more and practically the same for tee other three children. It is interesting that the largest percentage of the intake was excreted in the period following the greates intake with but one exception; the two periods coincide for V. B. In only two instances does the period of lowest percentage of intake excreted follow that of least intake. The children did not excrete the same average number of calories, but the percentage of total intake excreted was quite similar. The lowest average excretion was 110.4 - 67 - calories for J. H. and the highest 159.0 for E. 0. Al- though these amounts vary widely they both represent 9.5 per cent of the intake. 0. B. had a total excretion of 121.6 calories or 9.1 per cent of the intake, and V. B. 130.0 calories or 8.7 per cent. For children on a high protein diet want and her associates reported total out- puts representing 5.8 and 8.3 per cent of the intake (Table 4). These values are both lower than those in the present study. Output in feces: Individual fluctuations in the output of the feces did not vary directly with the variations in intake, because the percentage of the intake excreted in the feces was not the same for each period. The range was least (60.5 to 64.4 calories for J. H. and greatest (54.6 to 74.8 calories for O. B. In only two cases, for O. B. and V. B. did the periods of highest intake and greatest output coincide. The average total calorie output in the feces increased slightly with the intake for the first three children; 62.9, 66.2 and 67.9 calories for each respectively, while that for E. O. was much higher, 93.6 calories. This was nearly 26 calories above the output for any of the other children, and a much greater variation than had been found on the first study. Part of the wide variation in output in the feces for E. C. was, of course, due to his larger -58.. intake, but this does not show clearly except when expressed in term of the percentage of the intake excreted. E. C. had the largest percentage of excretion in the feCes, 5.6 per cent; J. H. slightly less, 5.4 per cent; 0. B. 5.0 per cent; V. B. least, or 4.5 per cent. The three children who served in both studies excreted the same percentage of the intake on both levels of protein. E. C. reacted like J. H. Individual variation from period to period was greater than that between the children. Since the subjects did not excrete the same percentage of the intake, the output in the feces did not vary directly with the calorie consumption. These fluctuations are comparable with those presented for the first study except for E. C. Wang and her associates, however, reported on excretion of 80 calories with a much lower percentage of intake excreted, or only 3.6 per cent. Another value of 36 was ecuivalent to 3.4 percent of the intake. In both cases the percentage of intake excreted was much lower than in the present study. Output in urine: Individual calorie outputs in the urine per period varied 11.0 calories for E. C. and less than that for the other three children. From period to period the values did not seem to hear any relationship to the intake. In only two cases, for J. H. and for V. B. did the periods of high output andhigh intake coincide, on the other hand, in no instance did the low values come at the same time. _ 59 - In every case less calories were excreted in the urine than in the feces, but, due to an increase in urinary calories the difference was less marked than on the medium protein diet. The increase amounted.to about a third of the number of calories excreted in the urine during the first study. The average value was 47.6 calories for J. H. and increased with the intake or size of the child so that it was 65.4 calories for E. C. According to Wang and her associates this increase was probably due to the larger amount of nitrogenous waste products such as urea and uric acid excreted by the children receiving the larger total protein intake. The fact that all of the children excreted practically the same percentage of the intake (4.1, 4.2, 4.1, and 3.9 per cent in order of their size) shows that the number of calories excreted in the urine was in direct progortion to the total mitrogen intake, and probably in- dependent of the calorie content of the diet as shown by the uniform increase for each child in the second study. Each child showed an increase of about 1.0 per cent of the intake excreted in the urine. This was the same as the percentage increase in total excretion for the high protein diet. These conclusions are comparable with those Wang and her associates formed from the results of a somewhat similar study. One of their subjects exereted 1.9 per cent of the intake on a medium protein diet and 2.2 per cent on a high - 70 - protein level. This was slightly less than the difference in percentages observed on the present experiment. Total absorption: Absorption fluctuated inversely with the fecal output as it did in the study in which the children received a medium protein diet. The greatest period to period variation (1527 to 1620 calories) was for E. C. and the least (1236 to 1299 calories) for C. B. Since the two diets were identical in calorie content and each child excreted approximately the same number of calories in the faces on the two studies the two average figures for absorption values for any child are very similar. The greatest difference was only 3 calories. On the high protein diet the lowest absorption value was 1105 calories or 94.6 per cent of the intake, and the highest 1573 or 94.4 per cent of the food consumed. Although the absorption increased with the larger intakes it remained relatively uniform for all the children as they absorbed the same percentage. V. B. had the highest average percentage of absorption (95.5 per cent) and also the largest absorption (96.0 per cent) for any one period. Likewise E. 0., who had the lowest individual figure, had the lowest average. These results are comparable to those Wang et al reported which were 96 and 97 per cent. They are, however, slightly lower. - 71 - Total retention: The retention remained.rather constant for each of the subjects, but varied with the intakes of the different children. There was a definite relationship existing between retention and intake for, without except— ion, the highest intake and the greatest retentian for all the children were in period 4 and the lowest in period 7. Individual fluctuation from period to period in calories retained was greatest (1462 to 1555 calories) for E. 0., and relatively uniform, though less, for the other three children. Because retention did vary directly with the intake and inversely with the total output, J. H. had the lowest average retention (1058 calories) and E. C. the highest(150$ calories) Since the calorie intake and the output in the faces for each child was practically the same on both diets, any dif- ference in retention on the two studies would be due to the increased number of calories excreted in the urine on the high protein diet. Average values (Tables 13 and 16) show that in this study the retention values were from 12 to 19 calories less than on the other study. This difference varied directly with the size of the subject, and, therefore, with the total intake. The variation was greatest for V. B., being 91.3 per cent of the intake on the high protein diet and 92.7 per cent on the medium protein level. The lowest average percentage of retention was 90.5 per cent - 72 - for E. C. Wang and her associates reported retention values, 94 and 92 per cent of the intake, slightly higher than those in this study. They found that both children retained 93 per cent of the intake in a medium protein diet. Study Number 11 High Protein Diet-On the Per Kilogram Basis Intake: Although the calorie intake on the high protein diet was quite constant, yet, when the values were expressed in terms of kilograms there was a variation from period to period. The greatest difference between the lowest and the highest intake was 5.32 calories for C. B. Fluctuations for the other subjects were only slightly less. In every case the children consumed the smallest number of calories per kilo in period 7 and the largest number in period 4. As in the medium protein diet, fluctuations were not the same for each of the children, and actually varied somewhat from child to child Fluctuations in the average intakes for the various children were slightly greater than those for a single child from period to period. They were also less than could be secured in studies in which the food was freely selected. The average intakes per kilogram of body weight in order of increasing size were $4.59 calories for J. H., 90.31 for C. B., 87.93 for V. B. and 91.17 for E. C. These values compare favorably with those reorted an the first Table 17 Daily Caloric Balances per Kilogram of Body Height on a High Protein Diet 3 3.75 -‘ v.8 1 1 .0 8 .48 . 1 . 1 .60 8 . 6 2 17.02 San $.23 3.87161 83.51 a 17.05 87.13 7.76 4.03 3.73 83.10 17.05 90.59 8 40 4.6 3.76 85.95 2 17 0 86.27 7.28 4.11 3. 7 82.15 17 1 88.63 7. 7 3.93 3.54 84.70 7 , 17.05 85.62 7.51 3.79 3.72 81.83 8 17.05 89.39 7.23 3.56 3. 6 85.83 var ' .' 3.'" 3.6 ' .° E'c' 8 8 6 8 n 8 88 1 1 2 0. .0 . . 0 . 2 1828 31.13 3.73 1.32336 52.53 a 18 28 90.27 8.89 4.99 3.90 85.28 18 28 93.86 8.81 5.22 3.59 88.64 2 18 28 89 8 8.85 5.52 3.33 83.82 18 28 92. 2 9.00 5.38 3.63 87.0 7 18 28 88.71 8.72 2.12 3.57 83.56 8 18 28 92.62 8.53 .9 3.57 87.66 '. .12 3. ' ., - ‘3.’ - 74 - study. There was a variation of only about one calorie between the intakes per kilogram an the two diets. These values were somewhat different from those which Wang and her associates reported from two subjects who received a diet high in protein content. The four year old consumed less calories per kilo (76.3 calories) and the five year old more (122.4 calories) than the children in the present study. Total output: The output per kilogram of body weight varied in much the same way as did the total calorie output pre- viously discussed. Individual fluctuations were least (7.80 to 8.22 calories) for'J. H. As discussed on the basis of total calories, the periods of largest intake and great- est output per kilo correspond for J. H., C. B. and V. B. On the per kilogram basis the eriods of least intake and low- est output were also identical for J. H. and E. C. The correlation between diet calories and the total excreted was 0.48 for the entire high protein study. This was a great increase over the correlation of only 0.04 in the first part of the experiment. When the data for the two studies were treated together the correlation was 0.24. The lowest average output, 7.62 calories, was for V. B. and the highest, 8.70 for E. C. These values were considerably higher than those reported in the first study due to the increased excretion of the nitrogenous products. In the experiment conducted by Wang and her associates, - 75 _ the total calories excreted per kilo were less than the averages for this experiment, being 6.3 and 7.1 calories for the two children. Output in faces: The individual variation in calories ex- creted in the feces was very slight for each child. Maxi- mum fluctuations, from 3.72 to 5.05 calories per kilogram of body weight, were for 0. B. and the least for J. H. (4.35 to 4.64 calories). The average calorie output in the feces per kilogram of body weight was relatively uniform for the various sub- jects. E. 0. excreted the largest number of calories (5.12 calories) and V. B. the smallest number (3.98 calories). These average values were practically the same as those excreted on the medium protein diets, there being less than 0.3 calories difference between any two values. The above results were comparable to those reported by Fang at al for two normal contrats, one four and one five years of age. Average figures show that they excreted in the feces 2.6 and 4.4 calories per kilo respectively. The difference between the intakes for these children was 46.1 calories. Muller reported a loss in the feces slightly higher (5.9 calories per kilo) than any mentioned above. Output in urine: Although J. H. had the least variation in fecal output she had the greatest fluctuation in urinary excretion (3.16 to 3.87 calories). It is interesting that - 76 - V. B. showed an opposite condition; the greater variation being in fecal output and the least in urinary excretion (3.5K to 3.76 calories). This pairing of high and low values was probably accidental and indicated that there was no definite relationship. The average number of calories per kilogram of body weight which were excreted in the urine were 3.uu for J. H., 3.75 for G. B., 3.6% for V. B. and 3.58 for E. O. This marked similarity in average outputs in the urine was also present on the medium protein diet. Nevertheless, there was one outstanding difference between the results obtained on the two studies: those on the high protein diet were about one-third greater than those on the medium protein level. The subjects of Wang and her associates also ex- creted a larger number of calories by way of the urine when the protein content of the diet was increased, but, since the calories were not the same on the two levels of protein intake, the results can only indicate a trend. One of the children who consumed 4.3 grams of protein per kilo, excreted less in the urine (2.7 calories) than the subjects on the present study, the other excreted a comparable number (3.7 calories). Absorption: The absorption of calories per kilogram of body weight varied directly with the per kilo intake and inversely with the fecal output. These variations were slight, how- —77-. ever, and were approximately the same for each child. E. 0. showed the widest range (83.56 to 88.64 calories( and also the largest absorption for any period. There was a high correlation (Table 15) of 0.95 when data from all the subjects on the high protein diet were used. This was about the same as that on the first study, 0.94, and slightly less than that on the two studies combined (0.97). For three periods J. H. absorbed less than 80 calories per kilo which helped to make her average the lowest (80.03' calories) for this study. E. 0. had the highest average absorption (86.06 calories). Since the calorie intake and the output in the feces were so nearly equal for both diets for any one child, the absorption values in the two studies were practically identical. The greatest difference was only one calorie. These results were less than the average -absorption value calculated from Muller's data (98.1 calories) and from Wang and her associates' data (118 calories). Another of the latter's subjects on a diet similar in protein content, but lower in calories, absorbed less, about 74 ' calories. Retention: When retention was expressed as calories per kilogram of body weight it varied directly with the intake and inversely with the total output per kilo for each child. Although this fluctuation in retention was slight, it was least (78.12 to 82.19 calories) for V. B. and most (79.99 to 85.05 calories) for E. 0. For every subject the highest -73- intake and the greatest retention per kilo were both in period 4 and the lowest in period 7. Thus the correlation between diet calories and retention was very high (0.95). when the data for the two studies were combined the correla- tion was 0.97 (Table 15). There was only a slight variation in the average re- tention values for the children. Both the intake and the retention in calories per kilo were less for V. B. than for C. B., although the average total calorie intake had been greater for V. B. than for C. B. (Table 16). This apparent discrepancy was due to the slightly greater calorie intake per kilo of C. B., who was not as much over the average weight as was V. B. The average retention per kilogram of body weight was least for J. H. (76.59 calories) and greatest for E. C. (82.48 calories). Since there were more calories excreted in the urine on this experiment than there were when a medium protein diet was given the retention values were ldwer. In every case the decrease amounted to about two calories. This decrease was proportional to the in- crease in nitrogen excretion, particularly that in the urine. I The data reported by wang and her associates show that the retention values for their subjects were either somewhat lower or a great deal higher than those obtained in the present study. Due to the wide variations in the number of - 79 - calories consumed per kilo by their subjects the retention values on the high and those on the low protein intake cannot be accurately compared on the per kilo basis. Period Averages Since diet calories in the present study were averages of two samples the average of the retention value of each of the three children on one period might be more comparable than the individual data. The averages for intake and retention which are dis- cussed in this section are different from any other presented in the experiment. They were computed by adding the intakes, then the retentione of the children for each period separately and dividing by the number of children. This gave the average intake and retention per period per kilogram of body weight. Table 18 and Chart 4 show the variations from the average retention per period per kilogram of body weight, and compares these with the average intake per period per kilo. When the retention and intake curve were plotted, they were nearly identical in contour. The individual values for each child are also marked on the chart. The table gives the calorie variation of each of these figures above and below the average. The values for J. H. varied more from the average on the high protein diet than they did on the - 80 - Table 18 Variations from the Average Retention per Period Per Kilogram of Body Weight Av. Intake Av. Reten.¢ V”ri t on fr m v . Puma pgr kg, per kg, Iinj::f""c'%, ',"Fv',s, §;§; _ H.P. 1 90.90 84.24 +2.95 +2.58 +0.36 2 87.3 80.51 +2.98 +2.9 0.00 a 88. . 81.52 -3.06 +3.0 89.50 81.96 -2.75 +2.14 2 87.52 80.23 -2.78 42. 6 +0.12 88.50 81.57 -2.88 +2.87 +0.01 7 86.83 80.03 -3.11 +2.74 +0.37 H.P. . 1 88.42 80.54 -3.23 +1.58 -0. 8 +2.0u 2 89.15 81.11 -3.49 +2.301-0. 9 +1.66 3 87.61 79.38 -R.67 +1.67 -0.02 +2.00 90.92 82.37 - .10 +1.60 -O.18 +2.68 2 86 75 78. 7 -3.79 .+1.24-+o.21 +2.05 89.44 81.2% -3.86 +1.69 -0.05 +2.21 7 85.93 77.9 — .91 +1. 1 +0.22 +2.09 8 89.78 81.93 -E.l4 +1.74 +0.24 +2.16 medium protein diet, while those for G. B. varied less. The fluctuations in the data for V. B. were practically the same on the two diets, but those on the medium protein were above and those on the high protein below the average values. The values for E. G. on the high protein diet varied to about the same extent as those for 0. B. on the medium protein diet. These values, of course, were proportional to the in— take figures and in each case followed the average curve quite closely. Nevertheless the average for all the children per kilogram followed the average intake curve more closely. some eaeeeum swam seen edeeeem sense: N m n H escapee N m m H c e 0 AV 0 a o o o o o , e \\@W// \\37/ MF/ 0 o < A” /..\ v. A/\ n e + + e m e, > . n > > > .1. > e 2.8 o > > > I . . ommmo¢¢ nll acapcuuom .\\ao«psepom .M.M + //// a r/ \\>/ \f/\ emoo > <\ \ \ /\ a... . vacuum emecH NN mN ow mm mm unwaok huom no amnwoadm Hod magnum nod soapsevom oweuo>4 as» sea“ anodumdnm> : enema mm setxoteo -82.. Discussion In the experiment the only appreciable difference between the two studies was the increase in the protein content of the diet in the second part. The average analyzed calorie content of the medium and the high protein diets were practically identical. There were, however, slight variations from period to period in the intakes for each child. The total output did not fluctuate in proportion to the intake variations on each period. This was partially due to the fact that the feces calories remained nearly constant for each child from period to period on both diets, and constituted about two—thirds of the total output. Although the per cent of fecal output remained the same on each period, J. H., who had rather loose stools, had the highest percentage output while V. B., who had rather constipated stools, had the lowest percentage output. Therefore the calories in the feces were influenced more by the character of the stools than by the nitrogen content of the diet. On the other hand the urinary excretion varied in pro— portion to the intake on each period. Furthermore, the nitrogen content of the diet affected the urine calories, as there was an increase of about one per cent in the per- centage excretion in the urine, on the high protein diet. This increase was the same for all of the children. - a3 - In all cases the absorption values varied inversely with the fecal output, and the retention with the total output. Therefore, the absorption figures were relatively constant for the various children and practically the same for both diets. Although the retention values were also uniform on each diet, they were slightly less, about one per cent, on the high protein level. This decrease was due to the increased urinary excretion. All of the data seemed to indicate that the protein content of the diet had prac- tically no influence on the calorie exchange. Summary 1. This investigation was made to study the affects of varying the nitrogen intake on the calorie balances of four normal pre—school children. 2. The average total calorie content of the analyzed diets for both studies were identical. 3. The only significant difference between the two diets was that on the medium protein diet the subjects con- sumed three grams of protein per kilogram of body weight, while on the high protein intake they received four grams per kilo. 4. There was a slight variation in intake from period to period. The greatest difference between the lowest and the highest for any one child was 94 calories for E. C. on _ sh _ the high protein diet. 5. 0n the medium protein diet average intakes ranged from 1169 to 1499 calories in total intake, or from 85.70 to 91.45 calories per kilogram of body weight. 0n the high protein diet average intakes ranged from 1168 to 1667 calories, in total intake, or from 84.59 to 91.17 calories per kilo. 6. There was an increase of about one per cent of the intake excreted on the second study. The least difference between the average outputs for any one child on the two studies was 11.9 calories for J. H. and the greatest 21.0 calories for V. B. since the number of calories excreted in the feces was practically the same on both diets, this increase was due to the greater number of calories excreted, in the urine on the high protein diet. This increase was probably due almost entirely to the products of a greater protein metabolism on the second study. The total output did not vary with the intake. 0n the other hand the urinary excretion increased slightly with the intake. 7. In every case the absorption values varied inversely with the calorie output in the feces and thus remained rela- tively constant on both diets. The percentage of the intake absorbed varied from 94.7 to 95.7 per cent on the medium protein diet, and from 94.4 to 95.5 per cent on the high protein diet. - 85 - 8. Since the retention values varied inversely with the total output they were slightly less on the second study. This difference was about one per cent, and due to the increase in urinary excretion. The percentage of the intake retained varied from 91.6 to 92.7 per cent on the medium protein diet and from 90.5 to 91.3 on the high protein diet. 9. Apparently the protein content of the diet did not affect the calorie balances of the children other than the increase in urinary calories on the higher protein level. - 86 - BIBLIOGRAPHY l. Atwater, I. 0. and Langsworthy, C. E.: A digest of metabolism experiments. U. 8. Dept. of Agr. Exp. Sta. Bul. No. 45, 1898. 2. Atwater, W. 0.: Principles of nutrition and nutritive value of food. U. 8. Dept. Agr. Farmer's Bul. 142, 1902 3. Bassett, Samuel H. and Van Alstine, Helen E.: Mineral Exchanges of man IV Variations in the mineral content of diets with a constant row weight formula J of nu- trition. Vol. 9 No. 2 p. 175. 4. Bedale, E. E.: Energy expenditure and food requirements of children at school. Proc. Roy. Soc., London, 94B: 358-404, 1922-192}. 5. Benedict, F. G. and Fox, E. 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