‘ . A‘ V W“..: o oh “.0.“ GI o H o .\. I “Ouch“ . 6 . \C.\ .‘ ah . om I" .u o . .0 a «u . C .\. o. a mu 0.. . a n“ 1" .. 5;; .. q. rlk '..\a T. . .v . w. 3 A .v .. "H any \ .. z .. ...\. .r . woo.“ ‘ I... H mm o. .. PJV‘O m. o \A. .\.- .. "a (“ . n .5 cl. . ‘J.’ n. lb .5. . u u ' n O a. b 0 \.\ 3.. ~ 3 cu - Z 06 f .0.- 0s . . .5 '3‘, a ’ 0‘ _ \ 7 .I u“ x . ”0:... 9‘. g 9 -\o . .J a o. . . x f . . r... ‘\.\ I .\ at‘Nd. - I. h‘ In”. K V a? ‘ I. a!” “I. t . “a . . . \ . n O” '. II .. \s. C. u A v. Q5 b . o.\. “a b 0‘. n o. a 1‘1. rm '...'5 U .\ I \ 91“ I w.” .W Va WW I \o. 3.. .. \ I. t t \ \-. a; Q. '..‘ - U _:__ TH 33!?! | This is to certify that the thesis entitled "A Calcium.Retention Study of an Eleven . and One-half Year Old Boy on a Self ‘ Selected Diet" ‘ presented by Alta V. Presson has been accepted towards fulfillment 1’ . I of the requirements for ' _H.:_5_c_ degree in Mutrition K . l 3 ]or professor f Date My 21: 1951 D .1 0-169 L n - -:.P a ‘1 fi Yv —Y ,— 17w" V'V . T Q :W‘V V r 1 —-l I \- 'a ” ’ 1‘ ' ' ' ‘ ‘ \ 4. x ' J l r . '. a ‘ . ‘ 1* ' \ ' O Q. I, h“. {I Q“ " ' l ' ‘ , ¢ ‘ ‘ J ' l a ‘ | ’ J. " \ (l ‘ I}? ' l ‘1',“ l I J I, ‘ l ‘ 1' l' l I g . r , I . ,- -I' 4 1/ .-, "H ‘ ‘ 1 I | i J _ I I I 4‘ . in y \ ., ‘ ‘ - I .,.‘ w v I 1 ' H . I I , 'f‘. ll . f j ’ ‘ 1' . l n' .- 1‘ . l ‘ l ' I ‘ 1- ‘( ‘ . ' I n ‘ ‘ .4 ' . r i ( ‘l, . I . ‘ "‘7' s. v .1 ' ' ‘ ‘4' ‘ ‘ .9. 'V o b l. n u . ‘I I l d' ': .0 ' ‘ 4 ‘ I " , . f l l ‘ . ‘ ~ "I l I' ‘ . . ‘ k l 1 , (‘ ' l l L ' ' 1) p ' ‘ . .l ’ t ‘ 0‘ I ‘ fl 0 ‘ 4 ' ls I", > ' f f. « D ‘f ‘ ‘ . . I '. n '. J .1 . ‘ "I J ‘ ' r ,‘, .l " t 'V 1 fi ' I . . . t . I f ' I o, ' l ’( ' | ’1 I ‘ u‘ I ' - ‘ pl ‘ ' ’ l ’ l -' I ' '0 '. ' C . I. I I I ’ fl . ‘ ' ‘ ’ w . ' ' I , ' I «Q . It: . t 4 t a . Ax . f- '” .~.‘_Ao-l ' A CALCIUM RETENTION STUDY OF AN EIEVEI‘I AM) ONE-W YEAR OLD BOY ON A SELF'SELECTED DIET BY Alta‘Virginia Presson A.THESIS Submitted to the School of Graduate Studies of Michigan State College of.Agriculture and Applied Science in partial fulfillment of the requirements for the degree of MASTER OF‘SCIENCE Department of Foods and Nutrition School of Home Economics 1951 ACKNOWLEDGEMENT The author wishes to express her sincere thanks to Dr. margaret A. Ohlson for her guidance and constructive criticism, and for her able assistance by making the anthropometric measure- ments; to my son, Billy Presson, who willingly and c00peratively acted as my subject; and to Miss Lois Jackson and mrs. Hazel Amen for their assist- ance in the laboratory. ¢)., - .. 'a-foeslt; TABLE OF INTRODUCTION . . . . . . . . . . REVIEW OF LITERATURE . . . . . . EDCPERIZIETII'AL PROCEDURE . . . . . Collection and preparation of Collection and preparation of Collection and preparation of Wet ashing method . . . . . . Dry ashing method . . . . . . RESUDTS AND DISCUSSION . . . . . Results . . . . . . . . . . . Discussion of results . . . . .EMARY AND CONCLUSION’. . . . . LITERATURE CITED . . . . . . . . IKPPENLDIXoooooooooooo CONTENTS urine . food. . feces . 14 23 24 25 26 26 30 30 38 46 TABLES IRE-"BER T ITLE PAGE 1 CALC IUM RETENI‘ ION DATE mo TILE LITE-MERE cozgcnmcngc SUBJECTS OF 1-»mJ-mom50111-m 314D ADL)I£;SCHJ‘ZII EEGES O O O O O O O O O O O O O C O C 12 2 FOOD EITI E RECOI-H) FOR THE SUBJECT FOR JANUARY15, PERIOD III . . . . . . . . . . . . 20 3 CALCULaTED FOOD VALUES OF DAILY 1332le FOR THE SUBJECT FOR JAIWMEY 15, PERIOD III . . . . 2]. 4 CALCIUM II"? FOOD MID FECAL EKICREI‘IOI‘IS AS DETERLILJED BY WET AILD DRY nSh'IIdG IvE'lTi-TODS. . . . 28 5 THE IIuTAKE , OUTGO AI‘ID PETEI‘TI‘ ION OF C; -.LC RIM OF THE] SUBJECT ON A SELF-SELECTED DLE‘I' . . . . 31 6 IIICRELSE IN CERT}, IN AI‘I'I‘I‘Bx‘Or'OLETRIC rm "b - IIIENI‘S AS RE TED TO CILLC IULI RITEI‘T ICE} OF SUBJECT ON A SELF SELECTED DIET. . . . . . . . 32 '7 M‘II‘HROFOI-ETRIC LEISURE; SEE-ITS DURII‘JG THE GROWTH OF AN EARLY ADOLESCEIT BOY (see Appendix) 8 BASI'LL METABOLIC PfiTES, IEIOGLOBDI, AND IESCCRBIC ACID DETEI—UJZ‘u-J‘IONS OF SUBJECT ON A SELF SLECTB) DEEP. o o o o o o o o o o o o o o o o 33 9 THE AVEEILGE II‘lTiECE OF CALORIES , PROTE LN , AND ASCORBIC ACID OF SUBJECT ON SELF SELECTED BET O O O O O I O O O O O O O O O O O 0 O O O 34 FIGURES I AN X-RAY OF THE 'a‘JRISTS REID REDS OF TIE SUBJECT STIOI‘JEG TIE OS;E:IFICATION OF THE CADEJELBOI‘I-flsooooooooooooooooo 17 INI‘RODUCT ION INTRODUCTION Many studies have reported calcium retentions of subjects in early childhood and of adult age, but few workers have studied the early adolescent. Due to the lack of information concerning the retention, and therefore the requirement, an interpolated value for the daily requirement for calcium for the early adolescent has been recommended (Recommended Dietary.Allowances, 1948). The daily requirement of calcium may be determined by the use of two types of information. As pointed out in a review by Holmes (1945), this information involves metabolism.studies to show the quantity of calcium used by individuals, and records of measurement of growth and body composition to show where the calcium has been used. It is desirable that more data be provided from.which the dietary requirement of calcium.for early adolescent children, may be determined. The object of this study, then, is to supply additional information concerning the calcium retention and increment in body size of an early adolescent boy. REVIEW OF LITERATURE REVIEW OF LITELATUHE There are two principal determinants of the requirement for dietary calcium during the growth period (Holmes, 1945). These determinants are the rate of growth and change in composition of the skeleton and the percent of intestinal utilization of the mineral. Various means for obtaining data concerning these two determinants will be mentioned. Each has its disadvantages. - The chemical analysis of the skeleton at various ages provides pertinent information, but data from such studies are limited. X—ray techniques showing the new centers of ossification (Todd, 1938), and determining the bone density (Mack, 31 al., 1939), have demonstrated the value of that medium in determining skeletal growth and composition. However, a question arises as to the physiological advisability of periodically exposing children to the x-ray. Anthropometric measure- ments of the skeleton at regular intervals also provide data from.which calcium accretion may be estimated (Vernar and Todd, 1933, quoted by Holmes, 1945). The method for determini-g intestinal absorption is the balance or retention study. vBy using data from several balance periods, the percent of utilization may be calculated. Holmes' (1945) formula for the calculation of percent utilization of calcium follows: Ca. balance during period A - Ca. balance during period B Ca. intake during period A - Ca. intake during period B times 100 equals percent of utilization. This formula, requires two levels of intake within the range of positive balance. Period A represents a high calcium intake and period B, a low calcium.intake. Studies conducted by the use of the methods mentioned provide data from which the daily requirement of calcium.may be estimated. Since this paper is concerned with only one of the methods, the retention study, the literature reviewed will be confined to factors affecting the retention of calcium. First, the length of the experimental period was observed to affect the variability of the results (Porter-Levin, 1933-1934). The retentions for two healthy pre-school children on a balanced diet were observed for a period of sixty days, and from fifteen to twenty-one consecutive days were required before the maximum.range of retention was reached. Balanced studies are extremely time consuming and expensive so periods made as short as possible for reliable results are desired. The "degree of saturation" of the body tissues has been observed by several authors to affect the retention of calcium. In an early study by Daniels (1934), a difference in retention of calcium.was observed between children who had been well nourished and those who had been poorly nourished. The poorly nourished group retained more calcium. Fairbanks and Mitchell (1936), using rats as experimental animals concluded that the "retention of calcium.by growing animals under conditions of adequate nutrition measured the requirement of calcium only when the calcium stores had been saturated by appropriate pre- feeding". Nicolaysen (1943), also observed higher retentions for "unsaturated" subjects. He studied the calcium absorption in relation to the body saturation of that element in the rat. Two groups of rats were fed diets differing only in calcium content for four weeks, then both groups were given the larger amount of calcium per day for two weeks. The average retentions for these two weeks were higher for the group that had received less calcium previously. Although the majority of the studies seem to indicate that unsaturated tissues tend to induce an increase in calcium retention when the intake is more plentiful, Stearns, at 31., (1941), did not find this true. These authors used malnourished children and observed that the calcium retentions mere below the retentions for norval children, but gr dually increased as intake increased. They concluded that undernutrition following illness deScreased the efficiency of the intestinal mucosa for absorbing calcium. Daniels (1941) studied the calcium retentions for three skeletally retarded boys and expected reduced retentions when the tissues were saturated. However, the retentions remained high throughout the study. Never-the-less, as a result of these studies and others, many investigators have provided preliminary periods in their work to enable the subjects to adapt themselves to the experimental regimen, and to saturate the tissues if possible. For example, if the subject has been placed on a controlled diet and the imrediste response to the diet is not a v riable in the study, sufficient time should be allowed for complete adaptation of the subject to the new regimen. If the effect of change in the diet is the variable to be studied, than a preliminary period for adaptation is not necessary. Hunscher,_et.al., (1936), provided one quart of milk per day for three months to ten children in order to avoid any unfilled stores in the bodies, before a comparison of skeletal maturity of the children to the calcium storage was made. also to avoid the effects of possible previous deprivation of calcium, Stearns (1934), provided each child on her study with the experimental diet until the calcium retention remained approximately the same for two successive periods. Another factor influencing the calcium retention is the calcium intake. The quantity of calcium intake has been observed to affect the retention of that element within a certain range which varies for each individual. Johnston (1944), observed a threshhold below wiich no cclcium.retention occurred and a physiological ceiling, above which no increased retention was observed. In this study, the calcium retention in relation to calcium intake, increased as did the intak rntil a given level for each subject was reached. then the "physio- logical ceiling" was reached the retention dropped. ho ill effects were re orted as a result of the extra intake of calcium. \ Kinsman, g£_al., (1937), also observed an apparent physiological ceiling. Lith intakes of 370, 610 and 880 mg. of calcium per day, five pre-school girls showed a retention which indicated that the 610 mg. intake was more nearly Optimum. The retentions for 370 and 880 mg./day intake were lower than the retention for 610 mg./day.intake. However, no physiological ceiling was reported by Stearns (1934). She reported that for each child the amount of calcium retention increased with increased intake. The series of studies included 347 balance periods using 66 infants and children from 7 weeks to 12 years of age. The range in gm./day of calcium intake was not reported in this paper. McKay, 33.213, (1942), observed the affect of intake on retention using young women as subjects. One hundred twenty five women were used, and the intake was found to be significantly related to retention at all intakes studied. The range of intake for this group of women was from 0.322 to 2.323 gm./day, and the mean calcium intake was 0.941 gm. Hunscher, gt 31., (1933), observed children differing in their reSpective retentions on each calcium intake. The studies included twenty days, fourteen days, thirteen days and eleven days consecutive observation for each of four children, the first three of them ten years old. When the diet contained one gm. calcium per day the average retentions were 0.26, 0.31, 0.30 and 0.48 gm./day, but when the intakes of the first three children were increased to 1.9 gm./day, the average retentions were increased to 0.55, 0.64, and 0.67 respectively. Although the retentions varied, each increased with increased intake. To illustrate further the individual variations of retention a study by Hunscher (1936) is cited. She observed healthy children who were five to eight years old and showed no deficiency of calcium stores as determined by the x-ray. The subjects received equal intakes of calcium. The retentions per day over periods of from twenty-five to sixty-five days varied from 0.27 to 0.48 gm./child. The relationships of the intakes of other substances to the calcium retentions have been reported in various studies. The calcium and phosphorus retention ratioS,. the quantity of nitrogen ingested, and the addition of vitamins D or 0 seem to be the most important and will be discussed here. The effect of different intakes of protein has been studied by McCance and others. McCance (1942), suggested that "very little calcium would be absorbed if no protein or amino acids were in the diet", and reported results from metabolic studies on five healthy adults. The results showed that increasing protein intake increased the calcium.snd magnesium absorption. McCance in his review mentioned that Mellanby, in 1921, observed improvement in canine rickets by feeding lean meat. In 1940, Kempster, gt_§1., in determining the relation between the utilization of calcium in milk and the calcium in dicalcium phosphate, observed that dicalcium phOSphate was not superior to milk as a source of calcium. MCCance suggested as a reason, that "the better utilization of the calcium in the milk was due to the presence of protein". Stuart (1945), observed a definite correlation between the amount of protein in a motherb diet and the length of the baby at birth. However, the results from a study by Hawks, £3 21., (1942), on the interrelationship of calcium, phosphorous and nitrogen in metabolism of the pre-school child, indicated that increased protein had no effect on calcium.sbsorption or retention. Calcium.and phOSphorous are both found in the bony tissues and musculature, or soft tissues, but the ratios are different. During growth, when a subject retains more phosphorous than calcium, it is usually suggested that soft tissue has been formed, and, similarly when more calcium than phosphorous has been retained, bony tissue has been formed. A dietary ratio of one part of calcium to one part of phOSphorus has been found to give desirable retentions during periods of rapid growth (Hunscher, 1933). The subjects varied in age from.three to ten years. The retentions of calcium and phosphorous respectively for three of the subjects were 0.26, 0.31, and 0.30 gm. and 0.36, 0.40, and 0.35 gm. When the intakes of calcium.and phosphorous were increased but the intake ratio was kept about the same, the retentions of both elements were increased. It is known that vitamin D is required for the efficient use of calcium. Vitamin D induces an increased "net absorption" of calcium from the intestines, and so alters the body fluids that bone salts may be incorporated into growing bone. Johnston (1944), observed the calcium.retention of adolescent girls with a range of daily doses of vitamin D from 0 to 5000 I.U. When calcium.intake was given in quantities to produce no further increase in calcium retention, the addition of 650 I.U. of vitamin D resulted in the highest calcium.retention of the series. Johnston found that the addition of vitamin D would reduce the negativity of calcium balance and promote a positive balance, only if the intake of calcium were marginal; however he suggests that supplementary vitamin D during the accelerated growth in height during early adolescence would be com- parable to that period of growth during infancy. In areas where an abundance of sunlight is available and the habits of living are such that the body is regularly exposed the value of the ingestion of vitamin D after the period of infancy may be questioned. The effect of trepical sunlight on the develOpment of bones of children in Puerto Rico was studied byCEliot, 1933). The incidence of rickets was very low as determined by calcium and phosphorous blood tests and x-rays. The few cases of rickets found were in invalids or persons who had reasons for not spending time in the sunlight. The studies made by McKay (1943), and Greenberg (1939), used young women as subjects and reported no effect of vitamin D on calcium retention. But these subjects were past the rapid growth period and probably received enough vitamin D from the sunlight for preper utilization of the calcium intake. Vitamin C as a factor influencing calcium retention was studied by Mellon and Rosenblatt (1946). Two groups of rats in positive calcium balance were each fed a basic diet. Grapefruit juice was added for thirty-six days to the diet of one group of animals. When the carcasses were ashed, and the calcium determined, the result showed that the group fed grapefruit had a much higher calcium.content. As reported in Brenneman's Practice in Pediatrics, a vitamin C deficiency may cause a cessation of growth of the long bones, and the ends of the bones may mushroom.out and acquire a grotesque shape similar to the condition in rickets. Scurvy interferes with the mechanism for removal of calcified cartilage matrix and suppresses the formation of new trabeculae. Thyroid medication in small doses resulted in increased retention of calcium for children on a constant intake of food (Johnston, 1941). When the doses of thyroid were large the retention was decreased. The doses of thyroid when given to subjects whose basals were lower than average, tended to raise the metabolism, and during growth, if the metabolism were not stimulated beyond normal, calcium retention was increased (Johnston, 1939), Higher retentions were observed when the basals approached normal; above or below normal, retentions were decreased. It is believed that the pH of the intestine is the factor that influences the solubility of calcium and therefore the intestinal absorption of that element. The solubility is greater in an acid solution and less in alkali; therefore most of the absorption should take place in the upper segment of the small intestine. Some of the calcium, however, will continue through the alimentary tract and be excreted. Christiansen (1936), discussed the conditions governing calcium absorption and reduced them.to one main controlling agent, namely the intestinal pH. She observed a small amount of calcium excretion from the intestine and no active excretion into the intestine. adolph (1940), also studied the calcium excretion into the intestine. He observed the effect of calcium in the form of diborgluconate injected intravenously into rats on the calcium content of the intestinal tract, and no significant increase was noted. Jones (1942), investigated the effect of different substances on the pH of six portions of the intestine of the rat and the simultaneous calcification of the bone. A greater acidity was found in the lower ileium and a definitely more antirachitic action was observed when oleic acid was ingested than when sodium oleate was given. 'Vitamin D did not lower the pH beyond that of oleic acid, but a greater anti- rachritic action was noted. The effect of the sex hormone estrogen, administered in varying amounts between 12,000 and 36,000 units to apparently normal girls during adolescence (Johnston, 1941) was a decrease in calcium retention by five out of six subjects. The decrease was due to increased losses in both urine and faces. The administration of stilbestrol to one normal girl at puberty also resulted in a depressed calcium balance. Additional factors influencing calcium retention as pointed out by thance (1943), Handler (1947), Silberberg (1948), and others are depletion of the subject in one or more factors of the vitamin B complex, caloric restrictions, and seasonal and annual changes. McCance reported the retentions were higher for a group of children in the summer months than for other seasons, probably beCeuse more sunshine was available. Handler demonstrated in young rats that caloric restriction to one-half of ad libitum feeding, other nutrients being adequate, resulted in a complete cessation of both skeletal and generalized body growth. The plasma concentration of calcium was normal in all rats. For B complex depleted rats (Silberberg, 1948), there was a retardation and subsequent cessation of growth of cartilage and bone. With the return to a complete diet the skeletal growth was resumed and normal conditions were restored. The calcium retention is affected by the rate of excretion of that mineral from the body, mainly by way of the intestines and kidneys. By far the larger quantity is excreted in the feces, but the excretion in the urine is more significant (ficCance, 1942, and Knapp, 1947). mcCance (1942) after observing that urinary excretions of normal persons rise and fall with intestinal absorptions, concluded that specific excretions "might be used as an index of chance in amount of calcium absorption". _ 10 - Knapp (1947) did not observe any relationship between the retention and urinary excretion. She states that the quantity of urine calcium is dependent on an endogenous factor, presumably endocrine, and also on calcium intake per unit of weight. The urinary calcium expressed as percent of intake varies inversely with intake per kg. of weight, and is an exponential function of the latter. Among the few investigators who have made contributions to the calcium retention as affected by the preadolescent spurt of growth are Wang, Johnston, Stearns and Jeans, and Sherman and Hawley. Wang (1928 - 1936), in a series of studies, used twenty-three adolescent girls ranging in ages from eleven to fifteen years, but investigation revealed that these subjects were deficient in calcium stores. The average intake of calcium.was 1604 mg./day, and the average retention was 11 mg./kg. of body weight per day, or 417 mg. with a range of '79 to 823 mg./day. About eight percent of the calcium intake was eliminated through the kidneys and sixty-five percent was fecal loss. Johnston (1939 - 1950), in a number of investigations, studied the factors influencing calcium.retention during periods of growth. The subjects were girls, and the retentions were observed under conditions of the reinfection type of tuberculosis. .A variation of some intake factors was also studied. Some of the results of Johnston's work and other authors are recorded in Table l. The range of retentions feund by the workers indicated in.Tab1e l is from.0.070 to 0.825 gm./day. The greatest retention observed was for a normal subject on a mixed diet, and the least retention was for a mal- nourished child. Other data include retentions of subjects under various conditions such as added estrogen and thyroid medication. - 11 - CALCIUM RETEKTI SUBJECTS OF P Table 1 0E DATA FRCM THE LITERATURE CCKCERKIFG ADOLES EXT AED ADOLESCZTT AGES a ‘4 Q) -3 Q“e a g0 10 m («g .2112?) 23>. :3 tat; ttt; a m 313 $33'2 3: 3 m 00) 00) H: HQ «4:13 «449 S on ea 33 as as. am .5 3:03 #38 3.53 32’. 88.3? 8‘8 smo/day sm./day smo/day Sherman and Haley 11 yr. F 0.748 0.456 0.013 Normal subject (1922) 0.994 0.729 0.021 on "mixed diet" 1.273 0.782 0.022 1.794 0.825 0.033 1.262 0.636 0.018 1.015 0.544 0.015 0.741 0.403 0.011 12 yr. F 1.02? 0.406 Normal subject 9 mo. 1.047 0.473 on "mixed dist" 1.057 0.336 1.030 0.226 1.076 0.326 1.091 0.229 1.002 0.163 1.056 0.090 1.053 0.333 1.068 0.371 Wang 12 yr. F 1.364 0.529 Probably had (1936) 1.799 0.591 calcium de- 1.799 0.466 ficient stores Johnston 13 yr. F 0.570 No estrogen (1941) 0.299 Estrogen 0.433 No estrogen Johnston 13 yr. F 0.503 No estrogen (1941) 0.328 Estrogen 0.458 No estrogen - 12 _ Table 1 (continued) 9 {2'3 3 "‘ v m {133 m m g g '8) g a? H +9 +> H H "—4 «4 -u ‘H o #40) 5 m g-p S-P m -P S m o m o m -fl,M -H a -a c a oH-P : es we as as as. em .5 3? 2'3 .3 a 8 ii 8 3 8 i“. 32° 8 ‘3 gnu/day ark/day sun/day Johnston 12 yr. F 0.247 No thyroid (1941) 0.310 One grain thyroid 0.359 One grain thyroid 0.193 Two grains thyroid 11 yr. F 0.245 No thyroid 0.215 One grain thyroid 0.342 Two grains thyroid 0.346 One grain thyroid Stearns, 11 yr. 1.586 0.235 Malnourished it. £51.. a 1.952 0.288 children (1941) 1.927 0.169 were used 12 yr. 2.353 0.509 1.955 0.447 2.056 0.369 13 yr. 1.93? 0.179 3.252 0.554 3.325 0.700 2.288 0.070 EXPERIMENTAL PROCEDURE EICPERDEPI'aL PROCEDURE This study reports the calcium intake and retention of one male subject, eleven and one half years of age, from.November to March, 1950 - 1951. The diet was self selected. The experimental study contained a preliminary period and four sampling periods; the sampling periods were one week in length and approximately one month apart. Period one was the preliminary period and collections were made and analyzed only for practice. The results for period one are not recorded in this paper. At the beginning of the experimental period, an examination to establish the physical status of the subject was made t; a physicianl. The examination consisted of the evaluation of the weight and sub- cutaneous tissue in relation to height, and the condition of teeth, gums, and skin. .A patch test and chest x-ray for tuberculosis, a threat culture and an x-ray of the hands to determine the osseous development also were made. The subject weighed 32.9 kilograms and was 140.5 centimeters tall at the beginning of the study. The subject, described as the wiry type possessed very little excess subcutaneous tissue. The subcutaneous tissue present, however, was firm, muscular, and possessed "spring". The subject was very active and seemed to have an abundance of energy. The skin of the arms and face was clear, smooth, and moist; and the lips were pink, moist, and full. The condition of the oral cavity was 1 Dr. Edith H. Kent, Lansi 9, Michigan good at the time of the study. The teeth were clean, seemingly well formed, and all cavities were filled. The gums were pink, firm, and filled between the teeth well. However the six year molars contained very large fillings. Most of the temporary teeth had been badly decayed before they were extracted. No temporary teeth remained. The subject had received.anm1e quantities of milk throughout infancy and early childhood. The child was a breast fed baby, and had consumed between one and two quarts of milk per day for the last several years. Scars were present indicating a history of rickets. Protruding scapulas and the presence of slight chest headings were observed, however the x—ray of the wrists showed normal ossification to date. The radio- graphic report of the subject indicated that the development of the carpal bones was well within the normal limits, as measured by the Todd Table or any other standard of bone develOpment. A.picture of the x-ray is found in Figure 1. The subject lived in the south until one year before this study was made. Additional vitamin D had not been given since infancy, but extra doses of vitamin D were again started approx- imately two months before the experimental period. Results from the patch test and chest x-ray for tuberculosis were negative. Negative results also were reported for the throat culture. As a summarization, the physical condition of the subject may be considered as good. He appeared to be a healthy, well nourished, and contented child. Basal metabolism ratesz, hemoglobin, and plasma ascorbic acid determinations were made three times during the course of study. Blood 2 A Benedict - Roth metabolism apparatus was used. - 15 - Figure I. An x-ray of the wrists and hands of the subject showing the ossification of the carpal bones. -17... J- uples were taken from a tricked finger, and he: :oglobin was de termir cd colorimetrically using a photolometer5 and the method suggested by Waddell, Steinbock, Elvehjem, and Hart (1928). The micro-method of Farmer and Abt (1936) Les ‘sed for the ascorbic acid analysis of blood. Lnth“01 cmetric reasurencnts, taken once a month, wer- similar in technique to those described by Teredith (1935), Davenport (1927) and Hrdlicka (1939). The measurements4 aken are list ei be 10?: Standing height Sitting height I:1::i:mJ.'1 he: d length Vaximur read breadth Nose breadth FaxiL um length of pinna (ear) Breadth of pinna Hand length Hand breadth Foot length Foot breadth Girth of lower leg Chest depth Arm span 3 A Cenoo-Sheard “Intolo 2 ter was used. 4 r . 1 1 The Fald”in Tale: heisurin g Scales and So luare , a metal tape, aid as spreading and slidingc cal p; s were the Lustrilments used. ghrn possible the left side of the suijct was measured. Weight5 was taken with shoes removed, but under- wear and pants were kept on by the subject. Similar articles of clothing were worn at each weighing. Grip strength6 was determined using a hand grip dynomometer. The subject was instructed to take a firm.hold and exert the maximum strength possible without allowing the arm.to come in contact with the trunk or lower extremity. One trial for each hand was made and the record was taken in kilograms. The blood pressureV and the pulse rate8 was taken each time the subject was measured. A.number of the measurements were discontinued for the last periods. It was felt that the growth changes by these measurements were not signifi- cant for an experimental period of only four months. - The diet was well balanced and checked by reference to the "Basic 7". The menus were made each week in advance, and foods were chosen that were seasonal and easily prepared. A typical dietary record for one day is given in.Table 2. A.calculated analysis of that days dietary is found in Table 3. The subject ate from the family table, and the method of 5‘A Fairbanks Scales was used. 6 A Narragansett dynomometer was used. 7 A.Nbrcury'nonometer and cuff was used. 8 The pulse rate was determined as the sum of two consecutive half minutes. -19- Table 2 FOOD IKTAKE RECORD 333 THE SUBJECT FOR JAIUAEY 15, PERIOD III Meal Foods Eaten Weight in Grams Breakfast Orange halves 7O Cooked rolled oats 150 Oleomargarine 10 Sugar 1% Scrambled eggs 35 Enriched bread 25 Milk 300 Lunch ‘ Vegetable soup 185 Roast pork 5O Bread 55 Salad dressing 1 Baked custard llO Milk (+50 Cookie 10 Orange 230 ’Dinner Grilled hamburger 85 Bread lOO Celery 25 Lettuce 25 Corn 70 Milk 375 Oleomargarine 10 After Dinner Banana 88 Coca Cola 340 L Additional vitamin D equivalent to 312 I.U. was taken each day in tablet form. The tablets were manufactured by White Laboratories, Inc. - 20 — .. 0.0 3.0 00-0 .. 00.0 0.00.." 00.0 0.0a 0.000 3 - H-0 H0.0 H0.0 - H-0 0.0 0.0 00.0 0.0: odaoao ems-0 00.0 0.0-0 00.0 .1000 91.0 0.03 0.00.” 00.2 3.02 0% u n u .. 2.0a 00.0 0.0 N001 .20 0.00 «0% .. 3-H 00.0 0H0 - 004 0.00 00.0 «0.0 0.03 33 0-0 00.0 00.0 00.0 . 00.0 0.00 0.0% 3d p.00 0000 finals-0.00M .. 0.0 00.0 00.0 .. Win 002 0.000 0.3 040: 0% u . 00.0 00.0 0.03 5.0 0.00 0.04. 00.0 0.0: 33 u .. .. .. u .. .. - .. 0.0a “00.00 .. - .. - 0.000 .. 0.40 “.04: 3.0 0.0.: 32%qu .. 8.0 00.0 0.10 .. 004 H000 00.2 dad 0-00 300.0033 900a 0-0 00-0 3.0 0.03 0.0 0.00 0-Ns 0.0 0-00 gala-meg 0.3 04 00-0. .00-0 0003 3-0 0.30 0.00.00 M.0M 0.000 wad-s .ma .MS .MS .ME .D.H .wa .ma .ma .aw 000... 03.803 5932 .003 .030 4 .3» 0.0 m .00 .020 .30 000.0 \ H: BEE .00 age-4.0 00a 0 .300 aofihmbm Hwy Nah deHzH HAH¢Q ho smfibfldfi mock nmadqpoqdo m Manama Moopvsmm os500500hw< .ohdpasoflnw< mo pdesphmmon .m.D II omma .0mummenm .0emweoosm .smm II mcooh mo sofipwmomsoo * 00.000 00.00 00.0, 00.0 00.000: 00.00 00.0000 00.0000 00.000 00.0000 .000000 . u u - u u - u - 0.000 1003000000 00-0 00.0. 00.0 00.0 0.000 00.0 00.00 .20.: 00.0 0.00 000000 0.0. 00.0 00.0 00.0 10.000. 00.0 10-00 0.0 00.0 0.00 0000 0.0 00.0 00.0 .00.0 0.000 00.0 00.0 0.0 0.0 00.0 naaannaw 0.0 00.0 100.0 00.0 n 00.0 0.00 0.00 00.0 0.0 000000 .mE .ma .wa .wa .D.H .me .wa .wa .Em 0000 00000000 000002 .0000 .0000 .4 .000 00 m .00 .0000 .000 0000 HHH nonmm .00 0000000 000 AvmdsapcooV 0 .300 sunbmpm mma Mom mmmazH MHH¢Q ho *mHDH<> nooh nmamgpondu service was unchanged from.previous eating procedure. The plates were filled in the kitchen then brought to the table. The subject was served a portion of each food on the menu and was expected to consume all of it; additional portions were available if desired. Each of the four sampling periods began on Mbnday morning before breakfast. During these four balance periods, all food and liquids consumed by the subject were weighed9 and these weights recorded. Samples of food and excreta were collected during the balance periods. A description of the methods used in the collection and preparation of urine, feces, and food for sampling is given below. URINE The urine was collected in one liter, screw tOp brown bottles that contained five ml. of concentrated acetic acid as a preservative. The first day's collection excluded the first voiding of the bladder upon rising, but included the morning voiding twenty four hours later. The succeeding days collections were separated similarly. Each twenty four hours collection was measured in a graduated mixing column, and with distilled water, made to volumne easily divisible by five. The urine was mixed fifty times; samples were taken for creatine and creatinine (Folin, 1914); and an aliquot of one fifth of the total volume was saved for the composite. The composite was kept refrigerated in a brown bottle until the end of each balance period. 9 A Hansen scale was used. -23 _ After the last day of each balance period the urine composite was allowed to reach room temperature, then mixed thoroughly by inverting and rotating. The composite ready for sampling was equal to one fifth of the urine excreted in seven days. Samples were met ashed as described later. FOOD Samples of food similar to that served and equal to one fifth the weight of that consumed by the subject were collected after each meal. The food was weighed on wax paper or in small beakers, and washed with distilled water into large containers. The solid and liquid foods were kept separate for ease in later handling. Both containers were stored in a deep freeze until time for sampling. The food was removed from the freezer the night before sampling to allow for thawing. Thawed food was transferred to a blender10 cup, and enough thawed liquid was added to fill the cup slightly over one half full. Care was taken not to fill the cup too full since the food was apt to foam over. The cup was covered and allowed to blend for five minutes. After all food was blended and transferred quantitatively to as many two liter volumetric flasks as were needed, the foam was allowed to subside. The flasks were made to volume, mixed thoroughly, and were ready for sampling. The total contents of all the flasks represented one-fifth of a seven day food intake. Each flask was sampled separately for ashing. The sum of calcium content of the flask was the sample total. 10 A Waring blender was used. - 24 - ,1 71d The focal samples were collected in quart-size d pa: ffined line d caid- board containers and co rrine was used as a marker. 0 carmine capsule was taken before broa1:fast of the first day and another before breakfast on the day following the last twenty four hours of the balance period. The fecal samples containing the first dos e of ca rwi11e vere sepasated and that portion of the stool excreted before the color showed was discarded. The colored portion and the poi rtion that follov.ed were saved as well as the succeeding samples until the second dosage of carmine appeared. Any part of the stool containing color from the second capsule vas discarded. The samples were dxted and fro: .en until the comlosite was to be made. The thawed feces v.ere placed in a blender cnpu, covered with distilled vater, and blended f. or five minutes. The blended material was trans- ferred quantitatively to a two liter 'elametric flask, and after the foam had subsided, the fla sk was made to volume, and finally mixed. after being thorou3hly mix ed, t is solution I;as ready for sampling and repre- sented the total fecal excretion for seven days. Samples were taken for ashing. Triplicate samples ..ere t Jzen from ea1ch com_: osite (food, feces, urine) for ashing, and duplicate portions were frozen in twelve ounce pharmaceutical bottles for later sampling if necessary. An additional experimental problem in ashing techniques was performed. Both wet and the dry ashing of certain food and fecal composites were done. The methods of ushing as used by this laboratory at the present time are described below. Triplicate samples of the blended composites were pipetted into Erlenmeyer flasks and placed on a hot plate. Five ml. each of concen- trated sulfuric acid, nitric acid, and hydrogen peroxide were slowly added, in that order, to each sample. Additional quantities of one to two ml. of nitric acidvere added upon each appearance of charring. The samples were allowed to digest until no further charring was observed then heated for about twenty minutes. The contents of the flasxs were dissolved with a few ml. of a 1:4 hydrochloric acid solution and trans- ferred quantitatively to a volumetric flask.3ach fLask was made to a volume of one hundred ml., mixed, and poured into a paraffined stoppered six ounce bottle and kept for chemical analyses. DRY AoHIIfG Triplicate samples of the composites mere poured into separate weighed porcelain evaporating dishes and dried. The fecal samples were dried partially over a steam bath under a hood, then to constant weight under infra red lanes. The food samples were dried in an oven at 400 C. All samples were stirred often. After being sufficiently dried, the samples were weighed, scraped, ground, and stored in labeled bottles in a desiccator. Duplicate one gram samples of the food and duplicate one-half gram samples of the feces were ignited in silica dishes in a muffle furnace at approximately 500° F. When the ashes we"e white or an even gray in color, they were dissolved with a few ml. of a 1:4 hydrochloric acid solution and transferred quantitatively to a one-hundred ml. volumetric flask. The flask was made to volume, mixed, and the contents were poured - 25 - into a bottle for chemical analyses. The calcium content of all the ashed samples was determined using McCrudden's method (1911). The analysis consisted of precipitating the calcium as calcium.oxalate and titrating with standard potassium permanganate at 90° - 100° C. Reagent blanks were prepared for each period. The results in mg. calcium /day as determined from both types of ashes are compared in Table 4. Higher values were found from the dry ash. This indicated that part of the calcium was not recovered from.the wet ash, probably for one of two reasons. First, the organic matter might not have been completely destroyed; or, secondly, some of the hydrogen peroxide might have been left which interfered with the titration with permanganate. The higher values obtained from the dry ashing method were used in calculating the retentions of calcium. - 27 - [.‘Pqu'v-‘r Tqv :1".C"’\ '.'r-_.~’<~a:~ “71'“ VTAp-v- 1!!" harp-r: \ch IL~U A -1“ ‘Ju _' J ‘JJ Ix afikLJ—J . i1 ‘J—JVaHLIAI .L.—’. .Lg.‘ l 5.x} 3 : T\‘.“r7'1“.")“ T".m :Y 7"7‘31 9.1“ “T .'. .1: ‘f'fl T’TFL"""Q ..-.. ..—a.-~.AL4' ‘d-I‘O" JJ 01‘... 31-.54' Jud—.5 -.,g.--.\.f b-JJaLéAKJ‘ ~ .‘4 ‘ ‘ :._.~ r a, ' . H.“ v w.- A’— 1 qu Ierioos : Calcium IntJne Fecal nicretion of vd;Ci u I -- 1 ‘— _. .. - 1' -~ . v- ! Dry net bGVlktlll fry net Ervleiicn I f .1. n . 1, t'. ." J, . v ASL; Ash 1 11‘. Dry Ash Luz”. .1 , Mr; I, ’5‘ . “ii i -1 .— V ;;’" v‘ {-4 qr —~—: ‘ ”1+- 4‘.- I I‘ .7 *1 r V ‘.\ ‘ I l 5 N "',~_' r‘é. /k 8.-é).l Wu Iv gut.’ L C v "“4. /\_U_‘. la v 1' T ' n , y I v i . If) 9". "0 ‘1ij ’3 ’2‘ ‘ fnq 0C 11”,!) (3 1r: III lLI—‘u.b.rU.K/"i; $.lh.’2 $1.3 W—JHI'Q‘u-v/ a——‘./."Lu' L1. 1 a V" "‘Nu'} 0.: ' "rvf. 4 1 I 11 ’ "Vi-'1 '2 “on - " [Ax/”’04!” L L‘VI‘OGJ 1 "-’04 l{ 90.17 L/,-’*0-C’ PESUIEFS MID DISCUSSION EDCLEERIIIEJEAL IESULTS During the four balance periods the subject ingested 2.043, 2.021, 2.043 and 1.873 gm./day of calcium, and retained 0.519, 0.520, 0.194, and 0.371 gm./day respectively. These data are found in'Table 5. The calcium intake averaged 1.995 gm./day, and the retention ranged from 0.194 to 0.520 gm./day. Certain of the anthropometric measurements are recorded in Table 6. Only the measurements that seemed to indicate a significant change within the experimental time limit were related to the calcium retention in that table. a complete record of the mersurements may be found in the appendix Table 7. A total increase in height of two cm. for the three month period, and a total gain in weight of 1.7 kg. were observed. The increment for each period is given in Table 6. A record of the basal metabolic rates, hemoglobin and ascorbic acid determinations are found in Table 8. The results of all tests were within the ranges considered normal. The intake of certain food nutrients for each period of study is given in Table 9. Total calories, protein, and ascorbic acid were calculated. An average of 2369 calories, 97.4 gms. of protein, and 115 mg. of ascorbic acid were taken each day. DISCUSSION OF RESULTS The range of calcium retention of this subject was within the range found in the literature. Sherman (1922) reported retentions for two early adolescent girls that ranged from 0.090 to 0.825 gm./day. The - 30 _ D‘ " p .d v Q.- CF ~~dg I-wv ‘T'fi '1? r "1 -I--. b: Coot-V '1” W) t“‘"“ 9‘? -‘ ---4;-~~ I Q . "1:91 *‘Ith A N i . ouL‘J *o‘. Lop-J. _———. ...—.00..— ”O fl‘Afi'. S m.+ V.- Tl Ia ... n a K... 9.). r.... In“. C. ...u 0 —I/ nU o (a ....V 3. 1 .fi (0 . . .v O / 1-x _ . v - y n.... l ITP - a a n. .1 o . b ..LD Au L 0.» I ..A J’I“ Iii. Ill. I I :4 7. :4 . v.1 . O O r. a a J M/ a; a; _ m w --4:=-,41 _ a _ v. 6 , _ 3 re. 1 a A. _ I... Au 0 o _ a / “c/ «1.3 _ A. .. O .x i A... . A. ./ 1 r./ a; :J a..- n.‘ h a a o o 7 Q). n _ ,3). d.) all .1. 11 Kt a) o o 1; /3 n; .. fl 1‘ 1. «I... . n ~ one All 1;. O 0 We fly an be A. A; 91 on _ r. 0‘. I III --- -. A. C... .1. . o _ 1.. . o 3 _ lb» A../ . «i ’1", I; 0-0+ 04mm 0.0 o¢w~ «.0: 0 ”mm Ndm . 183 won 33 arse 113 TH «E .3 13 pooh Puma Apooonm N.NN MSNN he mm Jumw 1&3 . pooh who..." 5%an do- To 90+ 04. Na? he as Tad deem Con Andaman 0-0+ mdfl m.o+ TS N. 6+ 0.3 «in Add and £3 5.qu :49. m.NN go N.Nm N6... ”MN odm 383 $309 Mango oi... mama 4.9 mi.“ :12. :43 d4; 3-33 fldflnm “mane 30+ @me m6... VS macaw. 0.3 31$ A sea A 4R4? 3.1% 00.34 mcmm @301 31mm :me Adds dimwdmz w «0+ m 194 m . 9. 04.0.3 oi? W133 m «9.: =8 m puma poms acme puma paws poms paws ImuodH Ionswmoz 1323 ~ tongue: .0."qu .0568: $8998: 36% 862 sedan 3.2m Assamese qofiafiom gaoado .03 o." ..R 3 .05 a ..Q .3 .08 w ..E 3 .08 m in.» .3 owe. > R3 :3 HH ueoaumm - Baum QMBRSMm gm 4 no Bahmbm ho ZOHBEHM wsHondo 08 38% m4 mhhuufiumpma OHMBflfimoEmP/Q “3483.10 2H MdeMOZH 0 Spas -32- Table 8 BASAL METABOLIC RATES, HEHOGLOBIN, AND ASCORBIC ACID DETERMINATIONS OF SUBJECT ON.A SJLF SELECTED DIET Age Basal Metabolic Hemoglobin Plasma Ascorbic Rates :acid_ percent gun/100 m1 gm. [100 ml. 11 yr. 6 m0. +8.04 13.63 1.5 11 yr. 8 1110. -1100 15063 1.6 11 yr. 10 m0. “206 14.40 1.4 Table 9 THE AVERAGE IETAKE OF CALORIES, PROTEIN, AND ASCORBIC ACID OF SUBJECT 0N SELF SELECTED DIET Period Calories Protein Ascorbic acid NUmber/day gm./day mg./day;g II 2074.86 88.57 105.91 III 2469.53 98.13 115.89 IV 2635.87 98.23 111.30 'V 2294.42 104.67 126.72 Average 2368.67 97.40 ' 114.96 . . . ‘- w. A C‘»‘ .- .v ' "W a. u '\ ‘3‘ ' I ~ 'p ‘- I‘ '. 9‘ conditions under whicL ~LPvr5Q go e‘vei .rat rarfe oi retort 31s, here L// -l _. ‘ ‘5 ‘5. a .- ..l_ m 1A a '1 A‘ u naitia-r: or this tr" um; ( 4°’:) swerve-"3- ti..";t ‘- 7 ~- . a” : «v-‘I . ‘- r~ J— " . A. 4‘ «3‘ .- \«-_ ‘h l././\ ,f‘. ”/31 , A r: c - m‘g a U’n‘u {3.15.er L O V r bits C‘.'- 3‘: PC 5 1“." (“1:15 ‘1 ..r'./~.. k". '(\"-.4 to \J.:/JJ. \ TH. 2'51” . ‘o-e ' .-- ‘ . 7 ‘. . 1 ’ ! . cu r . '1 c' a : retenticns as re ortca b; ucanctcn (1,41) “*d 0 r “as lj—i ‘Cngeu A U C Q ‘ Q ‘ from 0.C70 to 0.790 an./L327. and inst art‘v botn extreme vslnes were recorded for the scre e“bj act. The s bjee a use; by these authors were " . . ‘ “‘5‘ v‘ 1 W ‘.~v-- .-\t a ‘ . 3 ”I. v 1%, g “ W't‘ not ConleHIQC in normal bu¢81pun concit r“. ins; ‘~“s 6 nr 1- . ‘ F 1 F‘ ‘D I . ‘Ac on -~~ “. 'I . ‘t r . 1 nourish d or leg t.6 re1nf«ctiun t,'e of t.berculoris. Cthcr v—r1.r es the intKLe pattern exce t for one .tahes were practically the care, the retentions "er 9 the same and for ~eriod is the int me W”? decrease}, the retention also decreased. '7“. : _ I .. -. a . ‘y‘ ~ §__ 0 .. .‘ i.- ' o C . '_I ‘3’ I. _ .nls vari'tien ;.tn the lath“: are in accoxrnnce 21th the res_lts 3“ r- N 7 I: 1 V' at ..r~ ”‘7‘ (V qt" 7‘ " r' " J”‘I\V‘ 0+ ”‘9‘ " Y‘, 3! fit“ “'7‘ t‘ OLJ ~el\ ekt D. 9 (($41-73 kb$eA~l<.-‘A’ -j‘L¢.-_ .J ‘-.’ L'zJ-yL IV AA‘ 3 NI. . II 14" ‘~ 6 ‘ ' ,- ‘ A A . It is not “:0vn “r. t;e 11*ere0 retention cc~rrci 1 e forr - - : i a ‘ .: 1:, .. . ‘ ' . ‘- . .‘ ‘ '|-.- ‘. - perioc. Tn srsuect a c a sliHat Cold c"rino that L?TIOC hit this .7: not de;med sufficient to terminate the study at that time. The reteati n icr " ‘~ ‘ . - f: \‘D" N ~n r. n er 1 \ ‘4 1 . 't‘ ' tires or t e periois 1 need tram ‘4 to 25 i,v._-ice.it cf the intahe. "'/ ‘ a : . .— ' _' . .q- . 1 ‘m, ., ... ..c‘ -11 “n inierwstinr rel. tion_;1n an obeerrod between the creeth o, 1-9 a“ -: t I. 1 .9. - . yea , ‘fi a V- ‘r- 1w .r- 1 sundect ant tne calciim retention. It u.a 3199?.01 that the s e eton of the subflrct m'de severfl ChaRPOG between peri.ds two and three [—3rn / m‘— " r- . I‘ t ‘ A! 5" ‘- .‘ ‘ l' ‘ (T la a). lne at ture incre and one cm.; the leg lsngta inC1€chQ - - g N 1‘ . ‘ a 1 ‘A 'J n 1_ ‘ - _ .1. u_ 0 Q C.7 cm.; tle h.1a Chengee; ans t.o chest up tn anc 0:4a tn inzr 'eea. _ L P‘ ca -: C. The retention for period three was 0.520 gm./day of calcium. For period four a retention of 0.194 ans. of calcium per day was recorded. Less change in the skeleton was observed, although the sub- ject gained 1.06 kg. of weight during this period. The stature increased only 0.5 cm. The subject probably increased in soft tissues. Again for period five, the e was an indication of an increase of soft tissue with a continuation of the slowed rate of skeletal change. The increase in leg girth of 0.6 cm. and the gain in weight of 0.74 kg. indicated growth in soft tissues. The skeletal changes were about the same as for period four. This rhythm of growth from.the skeleton to the soft tissue and back to the skeleton, are similar to the changes as recorded by heredith (1935). The excretion of calcium through the kidneys varied with the calcium retention. This relationship was similar to that reported in the literature by McCance, although McCance did not use children as subjects. -36- SUIII'LXRY AND CONCLUSIONS SUIQJPRY AND COI‘ICLUS IONS The calcium intake and excretion of a healthy adolescent boy were studied for an experimental period of approximately ‘four months. Certain anthropometric measurements also were made periodically. The rate of growth was compared with the retention of calcium. The calcium intakes for the four balance periods were 2.043, 2.021, 2.043 and 1.873 gm./day, and the retentions were 0.519, 0.520, 0.194 and 0.371 gm./day respectively. The total increase in stature for the subject was two cm.; the leg length was 1.5 cm.; chest breadth was 1.8 cm., and the increase in weight was 1.7 kg. The calcium retentions seemed to correlate with the rhythm of growth. LITEEE‘URE C ITED LITERATURE CITED Adolph, W. H., C. C. Liang 1940 Calcium in the Alimentary Tract of the Rat. J. Biol. Chem., 122: 517. Brenneman's Practice in Pediatrics, 1: 13. Christiansen, H. 1936 Investigations Into the Excretion of Calcium. Doctorate Thesis, University of Copenhagen. Nutr. Abs.,_§: 722. Daniels, A. L., M. K. Hutton, E. Knott, G. Everson, 0. Wright 1934 Relation of Ingestion of Milk to Calcium.Metabolism in Children. .Amer. J. Dis. of Child., 42; 499. Daniels, A. L. 1941 Relation of Calcium, Phosphorus, and Nitrogen Retentions to Growth and Osseous Development. Amer. J. Dis. of Child., 62: 279. Davenport, C. B. 1927 Guide to rhysical anthropometry and Anthroposcopy. Waverly Press, Baltimore, Md.,'U.S.A. Eliot, M. 1933 The Effect of'Tropical Sunlight on the Development of Bones of Children in Puerto Rico. U. S. Children's Bureau Publication, No. 217. Fairbanks, B. H., H. H. Mitchell 1936 The Relation Between Calcium Retention and the Store of Calcium in Its Body with Particular Reference to the Determination of Ca1cium.Reguirement. J. Nutr., _11: 511. Farmer, C. J., A. F. Abt 1936 Determination of Reduced Ascorbic Acid in Small Amounts of Blood. Proc. Soc. Exp. Biol. and Med.,.§4: 146. Folin, 0. 1914 On the Determination of Creatinine and Creatine. J. Biol. Chem., 12: 469. Greenberg, D. M. 1939 Mineral I-Zetabolism: Calcium, lingnesiwn, and Phosphorus. Annual Rev. of Biochem, 8: 1369. Handler, P., G. S. Baylin, R. H. Follis, Jr. 1947 The Effect of Caloric Restriction on Skeletal Growth. J. I-Jutr., __3__4: 677. Hawks, J. 13., M. Bray, M. 0. L'ilde, LI. Dye 1942 Interrolationship of Calcium, Phosphorus, and Nitrogen in Ixietabolism of Pre-School Child. J. Nutr., 33: 283. Holmes, J. 0. 1945 The Requirement for Calcium During Growth. Nutr. Abst. and Reviews, _]._4_: 579. Hrdlicka, A. 1939 Practical anthr pometry. The liistar Institute of Anatomy and Biology, Philadelphia, N. Y. Hunscher, H. A., F. Cope, A. Noll, I. G. Macy 1933 Calcium and Phosphorus Storage in Growing Children. J. Biol. Chem., 130: IV. Hunscher, H. A., F. C. Hummel, I. L'Iacy 1936 Variability of Metabolic Response of Different Children to Given Intake of Calcium. Proc. Soc. Exp. Biol. Iied., 35.: 189. Hunscher, H. A., F. C. Hutuuel, I. G. 1.2acy, T. W. Todd, C. C. Francis 1936 Is Skeletal Maturity Related to Calcium Storage in the Child? J. Biol. Chem., _l_1_9_: lii. Johnston, J. A., J. 'w'. Maroney 1939 Factors Affecting Retention of Nitrogen and Calcium in Period of Growth, II Effect of Thyroid on Calcium Retention. Amer. J. Dis. Child., 58: 1186. Johnston, J. A. 1941 Factors Influencing the Retention of Calcium and Nitrogen in Period of Growth. IV Effect of Estrogen. Amer. J. Dis. Child., _§_._2_: 708. V Further Evidence of An Anabolic Effect of Thyroid on Calcium Lietabolism. Amer. J. Dis. Child., _6_?:: 1172. Johnston, J. A. 1944 Factors Influencing the Retention of Calcium and Nitrogen in Period of Growth. VI The Calcium and Vitamin D Requirements of the Older Child. Amer. J. Dis. Child, _61: 265. Johnston, J. A. 1950 Calcium Retention by Six Adolescent Girls. J. Nutr., .41: 137. Jones, J. H. 1942 The Relation of pH of Intestinal Contents of Calcium and Phosphorus Utilization. J. Biol. Chem., 142: 557. Kempster, E., H. Breiter, R. Hills, B. McKay, M. Bernd, J. Outhouse 1940 Ulitization of Calcium of Di Calcium Phosphate by Children. J. Nutr., 39: 279. Kinsman, G., D. Sheldon, J. Outhouse, J. Hathway, M. Smith 1937 The Calcium Requirement of Children. J. Home 1410., _2_9_: 571. Knapp, E. L. 1947 Factors Influencing the Urinary Excretion of Calcium. I In Normal Persons. J. Clin. Invest., _2_6: 182. Hack, P. B., A. T. O'Brien, J. M. Smith, A. t. Bauman 1939 A Method for Estimating the Degree of Mineralization of Bones from Tracings of Roentgenograms. Science, _8_9_: 467. Mellon, M. G., R. S. Rosenblatt 1946 The Effect of Fresh Grapefruit Juice and Conmercially Canned Tomato Juice on Calcium Retention. J. Amer. Dietet. Assoc., _2_2: 673. (Nutr. abs” _]_._6_: 639 (1946). McCance, R. A., E. M. Widdowson 1942 The Significance of Urinary Calcium, Magnesium, and PhOSphorus. J. Physiol., 121: 350. ' McCance, R. A., E. M. Widdowson, H. Lehmann 1942 The Effect of Protein Intake on Absorption of Calcium and Magnesium. Biochem. J., 352: 686. McCance, R. A., E. M. Widdowson 1943 Seasonal and Annual Changes in Calcium Metabolism of Man. J. Physiol. , 102: 42. McCrudden, F. H. 1911 The Determination of Calcium in Presence of Magnesium and Phosphorus. Determination of Calcium in Urine. J. Biol. Chem., 10: 187. McKay, H., M. B. Patton, M. A. Ohlson, 3131;. 1942 Calcium, PhOSp-horus, and Nitrogen Metabolism of Young College Women. J. Nutr., _2_4: 367. McKay, H., M. B. Patton, M. S. Pittman, G. Stearns, N. Edelblute 1943 The Effect of Vitamin D on Calcium Retention. J. Nutr., 26: 153. Meredith, H. D. 1935 The Rhythm of Physical Growth. XI: No. 3. Univ. of Iowa. Nicolaysen, R. 1943 The Absorption of Calcium as a thction of Body Saturation with Calcium. Acta Physiol. Scand., _5_: 200. Nutr. Abs., _]._3: 368. Porter-Levin, T. 1933-1934 Calcium and Phosphorus Metabolism of Normal Pre-School Children. II Successive Balance Studies Showing the Range of Variation in Calcium and Phosphorus Storage. J. Amer. Dietet. Assoc., _9_: 22. Recommended Dietary Allowances 1948 National Research Council Reprint and Circular Series, No. 129. Washington D.C. Sherman, H. C., E. Hawley 1922 Calcium and Phosphorus Metabolism in Childhood. J. Biol. Chem., _53: 375. Silberberg, M., B. M. Levy, F. Younger 1948 Skeletal Changes in Growing B Complex Depleted Rats and Course of Repair. Proc. Soc. Exp. Biol. and Med., _§_7_: 185. stearns, G. 1934 The Retention of Calcium from Early Infancy to Adolescense. J. Biol. Chem., _1_0_5: DOCKIV of Proceedings lCKVIII. Stearns, G., P. C. Jeans, R. Catherwood, J. B. McKinley 1941 Diet in Malnutrition and Celiac Disease. J. Pediat., _1_8_: 12. Stuart, H. C. 1945 Fed. Proc., 4: No. 3, 271. Todd, T. M., 3231;. 1938 The Use of the Roentgenology in Assessing the Nutritional State. 16th Annual Conference of the Milbank Memorial Fund. Waddell, J., C. Elvehjem, H. Steinbock, E. B. Hart 1928 Iron in Nutrition. VI Iron Salts and Iron Containing Ash Extracts in the Correction of anemia. J. Biol. Chem., 21: 777. Wang, C. C. 1936 Metabolism of Adolescent Girls. IV Mineral Metabolism. Amer. J. Dis. Child., _5_2_: 41. APPENDDC A ‘Vm' ,“ 'qrn‘“ ! A 3‘. A:JLOP V‘LQLI‘IC IVIDXDT“ flY ADELE CL“ Table 7 v-v- 'TR’TS DLva-VG “5.4—! .L‘. T BOY THE GROWTH OF AN JL 1 yr. 8 mo. 11 yr. 9 mo. 11 yr. 10 mo. (k:.) . _ . .. . , .2. Maximum Arm Sean £cm,) 14b 0 J 144,2 1L2,4d__gl l4~hl_, §itting Height ~ . 7° 7 ram , ,_-__ a. 2 j _.__z3_._3m 71.5 - in Head_Lc m1 gthlficnii 184;, 18.3 18.2. 18-5 Head-.33 waith Lem.) late 1315 13.8 13-9 Morph olo ical Fa.ce ‘ ! Lenrth (c: L) 11-8 111:: 11.7 . Ph mys 'ologice 1 Face / Length cm. 17.3 J,“ 17.Q 1?.5 5.? Eregdth of Ear_£gm.) 3.§___J 3.Lt - Nose‘Hei“Lt (C3111 “-9 “-6 - I‘TQS Bree. L1 uh. LC I‘D-J)— 3.0 Hand Length (out) 14.3 15.3 15.9 Hand “re dth (83.1 6.7 22.7 lad. ...... m" C Foot Breadth (C5,) 5.1 7.9 7.9 0.1 _ 1 ' . Girth of Lower Loggflan) 28.2 ‘ 38.9 25.0 26.6 1 -“-----.. i 1 Chest 3re.c h (c ) 42.0 i 23.2 22.2 ZZLQ __ m n * l" 0 W 111" ’ 19 F l:- u;.aSt #33311 Ln“ , __ *0 - 01’ ‘4'. . _L‘QU Table 7 (con inued “'m1"7"f7“."“m’?7.“ 1.1T}: Q'W“'l"§“m fi'T‘QT‘""1 FIT"? ;".‘,:'“""“"_' ’T j“ \ ...bsVA thafié5‘d “-UV;»«-¢0“_-.~= .LJV-‘-.A.-.g .L-...J J1.vn ..“ V- ..-. TAT)" r, -"‘":‘N\":"'m ”Jr" —-4£'L&hl-J[ ..tDv'Hc—J.J~‘-‘ JVY 11 yr. 1 11 yr. 11 vr 11 yr. A39 7 mo. 8 mo. 9 mo. 10 mo. Grip Strength R 25.0 25.0 20.0 25.0 (k:.) L 2220 22.0 1510 1_20-0 Ld E )4 '0 H (D U) I. I (V: Kb 1’.) IL;\ Ta .0\ 9 I- <\ U'I 1:— +0 13 .1? (I) H C) -;‘~ {B C) . ' grements L cm pm 4 pm 4 gm ELI. w... N 0 E n3 "U ....m PU a. U mu ‘4' ‘II' lo'tl. 0.- . (,1 l.l .I la 'IIHII. A. bla's‘.‘. e .x‘u 1Hl|||||illl|||||t|||||||i|||L|H|N|l|l|||||||||||||||||H|H 1293 03175 591