A STUDY OF DIETARY INTAKES C}? ELDERLY WQMEN AND THEIR RELATSON TO SOME SERUM CONSYETUENTS Thesis fa: fl'ia ”MW of M. S. MICHEGAN STATE UNIVER$ITY Sist‘er M. Clare; Wivia ESQ-whim, Q. P. 1962. nl.v LIBRARY Michigan State . University ByS Current i nature of the a fietary and die great volume of .3059 Of the pre of elderly borne; Status. MlVe mm is hit served a dietary records records were Use Examnation of t '3 1, . mats, “finaly; :esterol dEtem; ABS TRACT A STUDY OF DIETARY INTAKES OF ELDERLY WOMEN AND THEIR RELATION TO SOME SERUM OONSTITUENTS By Sister M. Clare Olivia Beaubien, 0.P. Current interest has stimulated much research into the nature of the aging process and its causes. Many non- dietary and dietary factors have been considered, and a great volume of literature testifies to the fact. The pur- pose of the present study was to relate the dietary intakes of elderly women to their physical condition and nutritional status. Twelve women aged 58-90 living in a cooperative home in Detroit served as the subjects of the study. Sevenrday dietary records were kept in fall, winter and summer. These records were used to evaluate nutrient intakes. Physical examination of the heart, pulse and blood pressure were made, and height and weight were measured. Biochemical examination included hemoglobin, hematocrit, red and white blood cell counts, urinalyses, and serum lipoprotein, protein and cho- lesterol determinations. \- lean daily 1 hydrate, were 48 15. 37 and 48 pe Younger subjecti older subjects hi’jrate calorie fatty acids Via: Host oi t‘ 75 per cent of respectiVely. at least 75 Pl nukes were ascorbic Beic‘ vitamin A am in suzmer th; SUbject er Cent Ove ceived 81. S respectivel‘ intakes. no 8m 'afpertaenSiv a. Sister M. Clare Olivia Beaubien, O.P. Mean daily dietary intakes of protein, fat and carbo- hydrate, were 48, 56 and 162 gm., respectively, supplying 15, 37 and 48 per cent of total calories, in the same order. Younger subjects usually had higher protein intakes than older subjects who consumed a larger percentage of carbo- hydrate calories. The ratio of linoleic acid to saturated fatty acids was 0.18. Most of the subjects received less than 50 per cent and 75 per cent of the recommended allowance for calcium and iron, respectively. Intakes of other nutrients usually equalled at least 75 per cent of the recommended allowances. Lowest intakes were for calcium, iron, thiamin, riboflavin and ascorbic acid, and highest intakes were in calories, protein, vitamin A.and niacin. Dietary intakes were generally lower in summer than in fall and winter. Subjects within 10 per cent of normal weight, 10 to 20 per cent overweight and more than 20 per cent overweight re— ceived 81, 99 and 118 per cent of the recommended calories, respectively. General health was often reflected in dietary intakes. Two subjects had abnormal hearts, and 4 subjects were hypertensive. Hemoglobin values ranged from 12.32 to 15.68 gm. per 100 m1. of blood; hematocrit ranged from 37.50 to 45.50 per cent 3.64 to 5.52 C6 whose hemoglobi wants indica' azbjects. Electroph azean of 66 ; Absinglobu? analyses show teia, with a : Per cent alph Subjects. Tc from 105 to ; tent). Unesa 24 to 66 mg, There w. 'Detalipopro these relati Sister M. Clare Olivia Beaubien, 0.P. 46.50 per cent of blood volume; and red cells ranged from 3.64 to 5.52 cells per mm. of blood, except for two subjects whose hemogldbin and hematocrit concentrations and red cell counts indicated anemia. Urinalyses was normal for most subjects. Electrophoretic serum protein patterns were normal with a mean of 66 per cent albumin and 34 per cent globulin. Albuminzglobulin ratios ranged from 1.1 to 2.6. Lipoprotein analyses showed a great preponderance of the beta-lipro- tein, with a mean of 85.5 per cent beta-lipoprotein and 14.5 per cent alpha-lipoprotein. There was great variation among subjects. Total serum cholesterol concentrations ranged from 105 to 284 mg. per 100 m1. of serum (mean 206 mg. per cent). Unesterified cholesterol concentrations ranged from 24 to 66 mg. per cent (mean 46 mg. per cent). There were no correlations between serum cholesterol, beta-lipoprotein, hypertension and atherosclerosis, although these relationships existed in individual cases. A STUDY OF DIETARY INTAKES OF ELDERLY WOMEN AND THEIR RELATION TO SOME SERUM CONS TI TUENTS By Sister M. Clare Olivia Beaubien, 0.P. A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Foods and Nutrition 1962 Approved by: AX '1’ 114/ (e/( '( LZ¢4 Q: 44¢. ACKNOWLEDGEMENTS The author is sincerely grateful to Dr. Evelyn M. Jones for her constant assistance, guidance and encouragement. She gives thanks to Dr. James Feurig, M.D., Director of the Michigan State University Health Center, for his interest in this work and for making available the facilities of the Health Center; to Dr. Fredrick Swartz, M.D. of Lansing for his advice in the planning of the study; and to Dr. Ezra Lipkin, M.D. of Detroit, Michigan,for his interest in the project, and for the physical appraisals and blood collec- tion. Further thanks are due to Margaret Smith, Nancy Ferrar, Mary Morris, David Anderson and Matthew Zabik, food and nutrition technicians, for their assistance. The author is especially grateful to Frances McNamara and the other residents of St. Catherine Cooperative House, Detroit, Michigan,for their patience, cooperation and good will dur- ing the course of this study. ii 4.... . $323... $52.41...“ a TABLE OF CONTENTS LIST OF TABLES o o o “o s s o o o s o o o o s o s 0 LIST OF FIGURES o o o o s o o o o s o o o o o s o 0 LIST OF APPENDICES O O O O O O O O O O O O O O O O INTRODUCI‘ION o o o o o s o o o o s s o o s o s o s 0 REVIEW OF LITERATtJRE O O O O O O O O O O O O O O O 0 Aging and Atherosclerosis . . . . . . . . . . . Nature of the Disease .. . . . . . . . . . Incidence of Atherosclerosis . . . . . . . Lipids and Atherosclerosis . . . . . . . . . . Triglycerides O O O O O O O O O O O O O O PhOSphOliPidsooososoooooooo‘ ChOlGSterOlsssoosssosossss Fatty Acids . . . . . . . . . . . . . . . Lip0proteins . . . . . . . . . . . . . . . Structure . . . . . . . . . . . . . . Characterization . . . . . . . . . . PrOperties of lipoproteins . . . . . Normal concentrations of lip0proteins Factors in Atherogenesis . . . . . . . . . . . Non-dietary Factors . . . . . . . . . . . HereditYosssosoossssos Arterial anatomy . . . . . . . . . . Age and sex . . . . . . . . . . . . . Homonessoosssoossssoo obeSitYsoosooooosoooos Hypertension . . . . . . . . . . . . DietaryFaCtors sossssosoosso Restriction of fats . . . . . . . . . Degree of saturation of fats . . . . Restriction of calories . . . . . . . Amount and quality of protein . . . . Intakes Of Vitamins o o o o o s o s 0 iii pAGE . v .viii . ix . 1 . 3 . 3 . 3 . 4 . 7 . 7 . 8 . 8 . 10 . 10 . 10 . ll . 15 . 16 . l7 . 17 . 17 . 18 . 18 . 20 . 20 . 20 . 21 . 23 . 27 . 29 . 31 . 32 TABLE OF CONTENTS (CONT.) Dietary Studies of Elderly PeOple . . . . . . Factors Influencing Dietary Intakes . . Nutritional Status of Elderly People . . EXPERIMENTAL PROCEDURES . . . . . . . . . . . . . Subjects and Environment . . . . . . . . . . Dietary'Data . . . . . . . . . . . . . . . . Food Intakes . . . . . . . . . . . . . . Nutrient Intakes . . . . . . . . .1. . . Recommended Dietary Allowances . . . . . Physical and Biochemical Examinations . . . . Laboratory.Methods . . . . . . . . . . . . . Hematocrit and Hemoglobin Determinations Electrophoretic Methods . . . . . . . . General procedures . . . . . . . . Lipoprotein analysis . . . . . . . Protein analysis . . . . . . . . . Scanning the strips . . . . . . . . Cholesterol Determinations . . . . . . . RESULTS AND DISCUSSION . . . . . . . . . . . . . . Nutrient . Intakes O O O O O O O O O O O O O 0 Physical and Biochemical Examination . . . . SUMMARY.AND CONCLUSIONS . . . . . ; . . . . . . . mmmATIONS O O O O O O O O O O O O O ‘0 O O O O “TEEN“ CI Tm C O O O O O O O O O O O O O O '0 . 0 iv PAGE 33 33 34 37 37 41 41 43 44 47 48 48 49 49 50 so 51 51 54 54 74 91 95 96 LIST OF TABLES TABLE 1. Vital statistics of the subjects . . . . . . . . . 2. Typical menus for fall, winter and summer . . . . 3. Computed ideal body weights and recommended dietary allowances O O O O O O O O O C O O O O O O O O O O 4. Mean daily intakes of energy, protein, fat and carbOhYd rates 0 O O O O O O O O O C O O O O O O O 5. Mean daily intakes of fat and distribution of the component fatty acids . . . . . . . . . . . . . . 6. Mean daily intakes of calcium and iron . . . . . . 7. Mean daily intakes of vitamin A, thiamin, ribo- flavin, niacin and ascorbic acid ._. . . . . . . . 8. Seasonal mean intakes of nutrients . . . . . . . . 9. Seasonal and mean distribution of subjects consum- ing specified percentages of the recommended allowh ances of mentioned nutrients . . . . . . . . . . . 10. Heart, pulse and blood pressure data . . . . . . . ll. Hemoglobin and hematrocrit concentrations and red and White 131006 C311 counts 0 o s o s s o s s o- o 12. Physical and chemical examination of urine Speci- mens C C O O O O O O O O O O O O O O O O O O O O O 13. Microsc0pic examination of urine specimens . . . . 14. Mean per cents of albumin and g10bu1in in serum proteins and albumin:globulin ratios . . . . . . . PAGE 38 42 45 55 57 61 63 66 68 75 77 79 80 84 LIST OF TABLES (CONT.) TABLE 15. Mean per cents of alpha and beta-1ip0proteins and alphazbeta ratios . . . . . . . . . . . . . 16. Mean concentrations of total and unesterified serum cholesterol . . . . . . . . . . . . . . . 17. Adjustments and assumptions used in calculating nutrient intakes . . . . . . . . . . . . . . . 18. Daily and mean nutrient intakes of Subject 1 . 19. Daily and mean nutrient intakes of Subject 2 . 20. Daily and mean nutrient intakes of Subject 3 . 21. Daily and mean nutrient intakes of Subject 4 . 22. Daily and mean nutrient intakes of Subject 5 . 23. Daily and mean nutrient intakes of Subject 6 . 24. Daily and mean nutrient intakes of Subject 7 . 25. Daily and mean nutrient intakes of Subject 8 . 26. Daily and mean nutrient intakes of Subject 9 . 27. Daily and mean nutrient intakes of Subject 10 . 28. Daily and mean nutrient intakes of Subject 11 . 29. Daily and mean nutrient intakes of Subject 12 . 30. Weekly and mean percentages of the recommended allowances in the nutrient intakes. . . . . . . . 31. Hemogldbin, hematocrit, red and white blood cell count 8 O O O O O O O O O O O O O O O O O C O O O , vi PAGE 87 89 L108 109 1111 113 115 117 119 121 123 125 127 129 131 133 137 LIST OF TABLES ( CONT. ) TABLE PAGE 32. Distribution of fractions of serum proteins . . . . 139 33. Distribution of fractions of serum lipoproteins . . 141 34. Total and unesterified serum cholesterol concen- trations . . . . . . . . . . . . . . . . . . . . 143 l v.1 ‘i LIST OF FIGURES FIGURE 1. 6. Number of deaths in the United States from all causes, from cardiovascular lesions and heart disease, and from arteriosclerosis in 30 to past looyears--19590osooooosoossss'os Schematic representation of hypothetical rela- tionship between hypertension, intimal haemorr- hage and atherosclerosis . . . . . . . . . . . Percentages of protein, fat and carbohydrate calories in the diets in each decade of age . . Percentage of recommended caloric allowances in the mean intakes of subjects who were nor- mal weight or overweight . . . . . . . . . . . Typical electrOphoretic serum protein pattern (0.005m1.8erum)...........o... Variations in electr0phoretic lipoprotein curves (0.03 ml. serum) . . . . . . . . . . . . viii PAGE 22 59 73 83 85 1.: -' 11'; LIST OF APPENDICES APPENDIX PAGE A. Dietarydata................. 107 B. Laboratorydata ..............‘. 137 ix INTRODUCTION Aging is a modern problem. Because of the triumph of medicine over many infectious diseases which formerly caused relatively early death, increasingly more people reach the time in life when they are called 'aged'. For some, the aging process is accelerated, and for others, it is greatly retarded. Such questions as, I"What is the aging process?', I'What factors control aging?', ‘What is an index of aging?', constantly confront investigators. Obviously, the aging process is complex and has many facets which cannot be measured quantitatively. However, blood lipids can be determined, and these have been impli- cated ca degenerative processes, particularly atherosclero— sis. This disease of the vessel wall is characterized by the deposition of lipids and subsequent development of fi- brous plaques. Generally, the severity of the disease in- creases with age. Early researchers regarded cholesterol the major cause of atherosclerotic lesions. More recently, the large cholesterol-bearing lipoprotein molecules have been considered more significant than other serum consti- tuents. Certain dietary factors caused increases, others 1 a . 4. WM; athei a brought about decreases in concentrations of lipids, in- cluding cholesterol and lipoprotein. Investigations have involved many nutrients, levels of intake and interrela- tionships of dietary factors. Considerable research sug- gested the importance of non-dietary factors. Results have been interesting, but sometimes contradictory. Many ques- tions remain unanswered. The purposes of this study were to evaluate the diets. of elderly women: to assess their health status; and to determine concentrations of serum cholesterol and relative amounts of serum proteins and lipoproteins. From the in- formation gained, recommendations would be made to improve the nutrition of this and similar groups. REVIEW OF LITERATURE A considerable volume of literature dealing with old age and aging is appearing at the present time. This con- siders factors of dietary and non-dietary origins, and at- tempts to define factors which actually indicate aging. One such recognized factor is the condition known as atheroscler- sis. Asimnridéichemaclemsis maimem Atherosclerosis is the most frequently observed patho- logical alteration of the intima and subintima of arteries, particularly of the coronary arteries. In the initial stages, fats are deposited in streaks in the walls of the arteries. Holman et a1. (1) have gathered evidence these 'fatty streaks' persist in a potentially reversible form for 15 to 20 years. The fatty streak is a collection of lipids just below the endothelium and is the beginning of the atherosclerotic lesion. An overgrowth of the endothelium incorporates the deposits of lipids into the arterial intima. Fibrous tis- sue gradually grows over and about the margin of the fatty 3 streak (l) . Florey reported (2) the lesion containi:: extracellular and intercellular lipid. At the base of the intimal plaque, cholesterol crystals as well as considerable amounts of cal- cium were found in a semi-fluid mass of lipid. Vasculariza- tion, ulceration and hemorrhage into the plaque resulted from.mechanical stress and turbulence of blood flow. Gresham, Howard, and King (3) demonstrated the lipid accumulation in fibroblasts or intimal smooth muscle cells was the first event in the production of 1eSions. However, these workers observed predominantly fibrous lesions with sparse lipid deposits in the plaques from human subjects 13 to 24 years of age. Acid sulfated mucopolysaccharides were present in greater amounts in the plaques than in the adja- cent media. The presence of polysaccharide and protein sup- ported the view that collagen formation was taking place. These lesions or thickenings of the arterial wall inter- fere with blood flow. Therefore, atherosclerosis plays an important role in many of the functional impairments that accumulate with age. Unless an adequate blood supply is available for all cells, changes take place in the body (4). WEEW All humans are susceptible to atherosclerosis. In 'r 'r'i', '.. ”‘.. _ i . - . . , ,. — - ' 1" -‘. a '.' ,) _ J . .— -. ‘ » - - a ’ " l ,, , . - [I y , , . .p _ , 1 J . , ‘l , , 5 ‘ . \-~ 1 ) ' \ n I l a. ', ‘ .. . ' . o . 4 '. - "‘l‘\ .‘ ‘ .I ‘ . . . - » . ' ' a *-, ‘hc " ' a (I ~ . I , _. - A. . .'L. ,‘ o r— . , —- ‘~ _ ‘\ k4 I _ i ‘ _1 t ‘ .. . - , _' r~ ‘ ‘ 4 -- ‘ . } . , _ 3 a 1 .' . f ,- - - ,4 . ' i , ' 4 4 _ . .fl ,- ._. - . . O . . _ _ , ‘ '.. . - , , J - . _ . . . . . . " . . . _ 1" s , _ I J I O ‘ ' 4- n - - r e ‘ I ,1 4 . .) .. I -. u ' . . -- . -. , . I) . is {I , 9 4| ‘ l ' - , -. ( r ’7‘ . ' , a l ‘ . L ' . .' s t , - ._., . . . I ‘ - . k ‘ , a. v ’. a o ' 7‘. I I- . - .1‘ l .. '1 _, . . _ . " - gtf’ ‘¢ . I “.. ‘ ‘ \ . . . . - - » ,. . .. ‘ . a ) ‘ l v.’ “ < . - .. . . I a ‘ ‘ I .. . . ,- v ‘ I ’ .‘. - . ‘ ‘ .. 1 ~— _ p . v ‘0 e | _ ; w u_ , a-_ _ .. . , a ' r‘ , . . I , '. J _ . . \ _ ' l j) . J a ' _ ,~ . — v' I -. I I I. J . a ._J i A) ’ .1 ‘ .- autopsies of 1,400 persons, aged 1 to 40 years, from differ- ent geographical areas of the United States, fatty streaks were found in the intima of the aorta of every individual over 3 years of age. The aorta was the first area involved in the disease (1). Vital statistics reveal the seriousness of the athero- sclerosis problem and of the heart and vascular diseases associated with it. Sebrell listed (5) the causes of death in 1950. Per 100,000 population there were 356 deaths from heart disease, 140 from cancer, 104 from diseases of the central nervous system, and 119 from other causes including accidents. A document published by the United States Government (6), reported a total of 1,656,814 deaths for the year 1959. Diseases of the cardiovascular system, caused 898,336 deaths, including 474,143 from atherosclerotic heart disease and 68,159 from hypertension. In the United States, heart di- sease was listed (7) as the leading cause of death in all groups of individuals over 25 years of age, regardless of sex or race. This was true for every state except Alaska, where accidental death ranked first. Figure 1 shows the to- :tal' mindset of deaths from heart and related diseases in each decade for the year 1959. om ca memoumaomoflnopum Eonm ocm .mmmsuwmnmms cos nmmm on .mmmmmflo Dams: ocm mCOHmmH Hoasomm> IOHUHMU EOHM .mwmgmv HHM EOHM wwflwfim muwanCD $3.... CH m£#mmmv MO HOQEDZ H MHDWHM whammy mm mm mm mm mfi mm — H — u — Ola“ o H“ \a(.\H\\\. \O a\ \o\o \\ O Q .\\ \\\\ '\ \ \ \D 0 .\d \ QI/ .\\ \\ .\\ .\ /d\ D \ .\ \. .\ a, \ \ \ D mHmOHmHUmOHHmHHmN III.|..II. O mmmmmflo Damon ocm mcoflmwa Hmasomm>0Honu mmmsmo HHQ 000.0N 000.00 000.00H 000.0VH 000.00H 000.0NN °0N 844989 30 ML" MW The role of lipids in the pathogenesis of atherosclero- sis has received much attention in recent years. The ori- ginal oversimplified hypothesis of the influence of fat has been modified, but experimentation still points to the imp portance of lipids. It is no longer possible to consider fat as an entity; rather it is necessary to think of specific lipid compounds. Goldsmith (8) lists 4 main classes of serum lipids: triglycerides, phospholipids, free or ester- fied cholesterol, and free or esterfied fatty acids. Erislxsenides Triglycerides may be considered the common storage form of energy. Normally, they appear in the blood for only a few'hours after meals. If the concentration is high, the serum is turbid, because these compounds are not very soluble. Albrink (9) suggested that an error in the meta- bolism of triglycerides was the lipid abnormality operative in coronary artery disease. This worker quoted 175 mg: per cent as the normal fasting concentration of serum trigly- cerides. Seventy per cent of the coronary patients studied had concentrations above this. Elevations of serum trigly- cerides above 160 mg. per cent occurred in 5 per cent of the persons over 50, and in 85-90 per cent of the patients with coronary artery disease. Wield: Phospholipids are important structural components of the walls and the mitochondria of cells. The importance of phospholipid as an emulsifying agent in the blood will be discussed later. Chapin (10) found blood phospholipid con- centrations 162 to 254 mg. per cent, constituting 42 per cent of total lipid. Gertler, Garn and Lerman (ll) ob- tained 299 mg. per cent for normal controls and 316 mg. per cent for a coronary group. .Shclsstsrcl The function of cholesterol is poorly defined, but its general wide distribution in the body suggests its importance. Cholesterol is present and can be synthesized by every cell. Atherosclerotic lesions contain deposits of cholesteroL which may account for 60-70 per cent of the lipids in the fully developed lesion. There is a statistical relationship between serum cholesterol concentrations and coronary artery disease (12). Keys and Anderson (13) stated high blood cho- lesterol concentration indicated a marked tendency toward the development of atherosclerosis. Lawry et a1. (14) re- ported higher serum cholesterol concentrations in men with established myocardial infarctions than age-matched men without obvious disease. In the Framingham study, Dawber et a1. (15) proposed those men with elevated cholesterols (over 260 mg. per cent) were twice as apt to develop heart disease as the total group. Those with levels below 220 mg. per cent had less than half the risk. Extensive studies have been undertaken to establish normal values for serum cholesterol. Aldersburg and co- workers (16) studied 1200 healthy males and females aged 2 to 77 years. In 20 women, 58-62 years, the mean choles- terol value was 263.8 mg. per cent; in 14 women, aged 63-67 years, 259.9 mg. per cent; and in 3 women, 68—72 years, 241.8 mg. per cent. In a cooperative study, several laboratories (17) around the country determined serum cholesterol concentra- tions in 15,000 people. The mean for men rose from around 188 mg. per cent at age 18 to approximately 245 mg. per cent at age 55, after which it declined. At age 20, both men and women had comparable concentrations. In women there was little change until age 30. At 50 years of age the concentrations were higher for women than for men, and continued to rise. . Lawry et a1. (14) found average cholesterol concentra- tions of 245 mg. per 100 ml. in 1560 adults aged 45—49 years. Twenty women, aged 60-69 had mean cholesterol 10 concentrations of 263 mg. per 100 ml. of serum. Scarborough (12) listed cholesterol values of 279 mg. per cent for nor- mal patient group aged 60-69; and 260 mg. per cent for those a decade older. Those men with elevated serum cholesterol concentrations (above 260 mg. per cent) were twice as apt to develop heart disease as the total group. mm In normal fasted subjects, Fredrickson (18) obtained 300 mg. of fatty acids per 100 ml. of plasma. Roughly 80 per cent of these fatty acids were present in phospholipids or in simple esters of glycerol. The triglycerides predominated in the latter fraction. Another 15 per cent of the fatty acids were esterfied with cholesterol. The remainder circu- lated as free fatty acids bound to protein. Free and esterfied fatty acids are important sources of energy. The non-caloric functions of fatty acids have been studied. Since these lipids seem to have an indirect role in atherogenesis, fatty acids will be discussed as a dietary factor in a later section. L' ! . l§$§§g§3§§. - Lipids are insoluble in water, so they are transported attached to protein molecules. Plasma lipopro- tein fractions represent a class of related, though not 11 identical, molecules. The fractions are not homogeneus, but their physical and chemical properties and metabolic relationships indicate they represent distinct classes of related molecules (19). In an historical reveiw, Oncley et al. reported (20) that Macheboeuf isolated the first lipoprotein in 1929, and found it contained 23 per cent phospholipid and 18 per cent cholesterol. Gurd et al. found (21) liproproteins represented 3 to 5 per cent of the total plasma protein and 75 per cent of plasma lipids. Page gave (22) the molecular weight of lipo— protein as about 1,300,000 and the composition as 77 per cent lipid and 23 per cent protein. The molecules are suffi- ciently large to be visible with a dark field microsc0pe. Although the lipids have a great preponderance over the protein, the solubility of these lipoproteins is typical of plasma proteins rather than of the component lipids (20). In the molecule, the structural arrangement is such that the peptides and hydrOphilic ends are exposed, furnishing mo- tility to the lipids (23). The major lipids of the blood, including glyceryl esters, phospholipids and fatty acids, cholesterol and its esters, are transported in the form of these giant lipoprotein molecules (24). .ShliflstfiiiZBSign, - Ultracentrifugation and electro- phoresis are among the various methods that are used to 12 separate the lipoproteins of serum. Depending on the method of study, the same lipoprotein may be designated by more than one name. The ultracentrifuge of Svedburg is used to produce an intense centrifugal field in which fractions of serum are separated according to their flotation rates. The designa- tion Sf means Svedburg flgtatiOQIQQit and is a convenient way of expressing density or the position of the fraction in the tube after centrifugation. The Chylomicrons, which are extremely low density (Sf 0-12), form a creamy layer at the top of the tube.- They contain less than 1 per cent protein and 99 per cent lipid, and are important vehicles in lipid transport. Chylomicrons are found in serum after a meal containing fat, and produce lactescence in serum when present in large amounts (8). The beta-lipoprotein fraction low density (Sf 10-400 class) is the other important vehicle for the tranSport of fat. It can be found even in fasting blood samples. The beta-lipoproteins consist of lipid associated with serum globulins. Analyses showed the content is 21 per cent pro- tein, 23 per cent phospholipid, 47 per cent cholesterol and 9 per cent triglyceride, (25). This represented most of the serum cholesterol and a major portion of serum trigly- ceride. 13 In the opinion of Olson (26) beta-lipoproteins were the primary agents in atherogenesis, while other factors in the host or environment determined whether or not lesions would develop. Gofman et al. (27) suggested that Sf 12-20 lipo- proteins correlated better than other serum lipids with coro- nary disease. Subsequently, Gofman extended the range of atherogenic lipoproteins to the whole of the beta—lipopro- teins (Sf 0—400). Gofman observed these complexes deposited in atheromatous plaques and in high concentrations in the blood of persons susceptible to atherosclerosis. The high-density lipoproteins were composed of non- esterfied fatty acids attached to albumin. There was no evidence linking alpha—lipoprotein to atherogenesis (8). In the original electrophoresis cell, Tiselius applied (28) an electrical potential to each end of a U tube con- taining a solution of protein. The protein fraction mi- grated toward one of the electrodes. Blix, Tiselius and Svennson (29) separated lipoprotein by electrophoresis of the serum in a U tube. Modifications of the technique in- volved migration of the protein in an electrical field pass- ing through a solution supported by a material such as silica gel, potato starch or glass powder (30, 31). More recently, electrophoresis with filter paper strips has been introduced. Durrum pioneered (32) in this procedure, 14 applying an electrical potential across the ends of a paper strip saturated with buffer. A small sample of material was placed on the paper. After migration and staining, the op- tical densities of the stained serum fractions on the strip were read on a scanner. The relative amount of each protein fraction could be determined from the distinct colored zones which developed when electrophoretic paper strips were subjected to protec- lytic dye, such as bromphenol blue. The scanner was an absorptiometer type of instrument in which the optical den- sities of narrow sections of the strip could be read from one end to the other, and the readings plotted directly on a paper to give a graph of the concentrations of dye along the strip. Measuring the area included in each section of the curve allowed the worker to calculate the relative amount of protein contained in each fraction (33). The largest peak of the graph contained the albumin fraction, and the smaller fractions were the globulins. The ratio of albumin to glObulin was believed to be indica- tive of the health of the person. Koiw reported (34) ratios for healthy subjects in the range 1:1 to 2:1. Lipophylic dyes such as Sudan Black (35) or Oil Red 0 (36) have been used to stain the lipoprotein in the paper strips. When the dyed strips were scanned and the graphs 15 were plotted, 2 main peaks became visible. The first peak contained about 30 per cent of the area under the curve and represented the lipoprotein with the highest rate of migra- tion, the alpha-lipoprotein. The larger peak contained about 70 per cent of the area under the curve and represented the beta-lipoprotein fraction (35). Flynn and de Mayo (37) enumerated certain advantages of this method: the apparatus was simple, very small samples were needed and many analyses could be run simultaneously. This method has been used extensively in clinical and experi- mental work. W p; lipoproteins. - Water is a major com- ponent of the lipoprotein molecule; it has been estimated there are 44,000 moles of water per mole of lipoprotein. The importance of water structurally is suggested by the sensitivity of lipoproteins to freezing and drying, both of which produce irreversible changes in the molecule. The water of hydration seems involved in binding the polypoptides to the lipids. Besides the tendency to lose water, the lipoproteins have a marked susceptibility to oxidation. With the aging of serum samples, the beta-lipoproteins increase in electro- phoretic mobility. Therefore, serum should be used as soon 16 as possible, and if storage is necessary, it should be done in concentrated solution at 0°C. (39). Beta-lip0protein seemed more prone to deposition in sub- intimal tissues, or less stable, than alpha-lip0protein. The colloidal stability of lipoprotein complexes rather than the total concentration of any particular component of the molecule may prove to be the variable that determines whether or not lipids will be deposited (40). Phospholipids seemed to be this colloid stabilizer because it emulsifies lipopro- tein complexes to relatively small size (41). ‘When these complexes are smaller, they are less likely to be deposited in arterial intima. Individuals with atherosclerosis have a relatively high phospholipid-cholesterol ratio. The normal ratio is 1:2 (42, 43). Ema]. W 2: We. - Goldbloom used (44) paper electrOphoresis to determine serum.protein and lipoprotein in normal patients 80 to 94 years of age. About 60 per cent of the protein was in the albumin band and the rest was in the globulins. In normal patients, 60 per cent of the lipid was in the beta-lipOproteins: while in abnormal cases, as much as 80 per cent of the lipid was in the frac- tion. Eiber and associates found (45) no significant dif- ference in phospholipids and cholesterol, but total lipids wifwiiriqs'l‘,‘ ‘ |||la . ll|l 17 decreased after the age of 60 to 75 years. The normal pattern in the paper electrOphoresis reported by this author was 60 per cent of the lipids in the beta-lipoProtein and 40 per cent in the alpha-lipoprotein. In a series of normal plasmas, Oncley and coworkers found (20) approximately 75 per cent of the plasma lipid bound to the Beta-lipoprotein: which represented 5 per cent of the normal plasma protein. Lawry and coworkers found (14), in women aged 60 to 69, 71 per cent of total lipid occurred as beta-lipoprotein. mum WW Coronary atherosclerosis in a human being results from a number of causative factors. ,flgggditx. - The anatomy, physiology and biochemistry of the vessel walls is believed to be conditioned by heredity. This conclusion was reached because the clinical manifesta- tions of atherosclerosis and the conditions that precede it are often observed in all or nearly all members of a family. Wilkinson et a1. studied (46) a large farm family of 91 mem- bers spanning 4 generations. Hypercholesterolemia was present in so many of the family members in all ages that 18 these workers proposed that this characteristic is trans- mitted by the genes. ,Agtggigl anatomy. — Moreton theorized (47) the arterial anatomy was such that it predisposed to the deposition of lipoproteins. The intimal walls were formed in such a way as to trap foreign particles, and the large lipoproteins stimulated this response. Smaller molecules entering the vessel walls were readily reabsorbed into the blood, but large ones were trapped between the high pressure blood column on one side and the internal elastic membrane of the artery on the other. Thus, the lipoprotein particle was deposited and retained in the arterial wall. .§g§.§gg,§gx. - The development of atherosclerosis and the concentrations of the blood lipids and lipid-protein complexes which are often used as indices of the advance- ment of atherosclerotic damage appear to correlate with age and sex. Russ and others (48) found that the plasma of young women between the ages of 18 and 35 contained less beta-lipoprotein than that of men of comparable age. Other- wise, there was no significant difference in the lipid- protein distribution in the sexes. In a group of men and women aged 75-91, women revealed significantly higher lipid concentrations than men. In women, the serum lipid Ci 19 concentrations rose continually up to the age of 80: but in men, the rising trend ended at 55 years (49). Sperry and Webb compared (50) the cholesterol concentra- tions of blood serum for 14 men and 8 women with values found for the same subjects 13 to 15 years earlier. In 8 men and 1 woman, no appreciable change occurred, but in 6 men and 6 women, increases in serum cholesterol were found. Serum cholesterol concentrations and also lipoprotein content tended to increase with age, but the elevation did not necessarily parallel chronological age. Dawber et a1. included (15) that individuals in middle age (40 to 50 years) with elevated serum cholesterol concentrations had greater probabilities for de- veloping atherosclerosis than did older persons with comparable serum lipid concentrations. In a study of air force personnel, aged 20 to 40, Milch and coworkers found (51) twice as much serum.beta-lipoprotein in the 40-year-old group as in the 20-year-old group. In general, these workers believed, serum cholesterol and athero- sclerosis both rose with increasing chronological age. Mean values of low-density lipoproteins were demonstrated to be linear functions of chronological age. Standard deviation of these means increased in accordance with an expected increasing divergence of individuals aging at different rates. .‘w .r..i.(Fv'-1I§.7 L: e . 2O .flgzmgnefi. - Holman (1) found the most striking increase in the degree of the atherosclerotic condition occurred be- tween the ages of 8 and 18, and suggested a relationship be- tween atherogenesis and the hormonal changes of puberty. Cornwell et a1. (52) found hyperthyroid patients had elevated serum cholesterol, phospholipid and fatty acids, but not necessarily elevated lipoproteins. Furman et a1. (53) observed hyperlipemia associated with such disorders as dia- betes mellitus and hyperthyroidism. thfiiix. - Wilens compared (54) the incidence of various degrees of general and coronary atherosclerosis in obese and poorly nourished persons. Advanced atherosclerosis was about twice as common in the obese as in the poorly nourished. The persons of average nutrition were intermediate in the degree of atherosclerosis. This author stated the most striking factor in atherogenesis, especially of coronary arteries, was obesity; and that the condition was more dangerous in men than in women. ‘flypggtgngign. - In autopsies of hospital patients, Gofman et a1. (55) found appreciable high amounts of Sf 10-20 lipoproteins in 92-94 per cent of the arteries of hyperten- sive patients. Paterson and associates (56) believed hypertension 21 promoted atherosclerosis and theorized the mechanism in- volved was an increased tendency for intimal rupture and hemorrhage and consequent enhancement of atherosclerosis. These workers developed a schematic representation of the hypothetical relationship between hypertension, intimal hemorrhage and atherosclerosis. This is reproduced in Figure 2. I Sacks (57) compared the incidence of atherosclerosis in hypertensive and normotensive persons. Atherosclerosis was present in 80, 40 and 70 per cent of hypertensive men, and 23, 29 and 40 per cent of nonhypertensive men, in the fifth, sixth and seventh decades, respectively. This inves- tigator found atherosclerosis in 25 and 50 per cent of hyper- tensive women and in O and 17 per cent of non-hypertensive women in the fifth and sixth decades, respectively. Wm Various studies related dietary intakes to blood lipid and lipoprotein concentrations. Total fat content of the diet 3f the proportion of various fats seriously affected serum cholesterol concentrations in adults. No direct proof of a relationship between serum cholesterol and atherosclero- sis has been established for man. However, population stu- dies indicated a marked tendency toward the development of 22 HYPERTENSION transient persistent (from stress) V reflected into 1nt1ma1 capillaries a" 8 92a of early and older on atherosclerotic plaques o o g; ‘5' e e elevated pressure in ‘5 a 7' CY intimal capillaries ‘6 <3 (D Increased capillary \ 9n fra ilit " g Y\‘rupture of 1ntimal capillaries 7?; i 3 ‘3’, internal haemorrhage Haemosiderin and/ ceroid deposition Fibrosis and secondary from haemorrhage Arterial thrombosis vascularization from from injury from haemori'hage Acceleration of atherosclerosm FIGURE 2 organizatio/n of haemorrhage Lipid depos1t1on Schemat1c representation of hypothet1ca1 relat1onsh1p I I O sclerosis (56). between hypertensmn 1ntimal haemorrhage and athero— . In“! L.“ ._. ”I .s e! 1:2 . ’9‘ tr 23 atherosclerosis in individuals with high concentrations of cholesterol or cholesterol-bearing lipoproteins, (13). Bestgigtign.gf.§a§§‘ - The death rate from circulatory disease in Europe declined sharply during 1939-1945. This wartime period was characterized by limited consumption of milk, butter, cheese and eggs. In England, the disease was twice as prevalent in the professional and executive classes who were able to go to rural areas to obtain scarce food items, as in the laboring classes who did not have this oppor- tunity (58). During World War II and immediately following, the Scandinavian countries conducted statistical studies on diet and heart disease. In Sweden, death rates from atherosclero- sis rose until 1941, but fell during 1942 and 1943. This corresponded to the period of rationing of foodstuffs with accompanying reduction in the consumption of butter, eggs, and meat. The mortality rate declined more in the urban popula- tion than in the rural area where the rationed foods could eas- 111 be obtained. When the government ended food rationing, the incidence of heart disease rose again. In Finland, the mortality rate from atherosclerosis also fell during the war. After the termination of food rationing the incidence of heart disease increased again. In Denmark, where total fat 24 intake declined and butter and egg consumption rose, the athersclerotic death rate remained constant. Despite the rationing of fats in the United States, coronary disease did not decrease, possibly because the consumption of eggs and dairy products, except butter, increased constantly (59). In certain parts of Russia, Anderson Observed (60) many old people who ate 3 or 4 meals a day,but avoided overeating. Their dietary of vegetables, dairy products, cheese and yo- gurt slowed the advance of senility. Malmros reported (59) atherosclerosis is a serious problem in other parts of Russia where pe0p1e are in the habit of consuming enormous quantities of fermented mare's milk. The disease does occur in China where the diet is mostly vegetarian, but the Chinese do not suffer the wide- spread lesions of Americans and Europeans. In a village for the aged in Israel, Toor et a1. com- pared (61) the diets of European and Yemenite immigrants. The Eur0peans consumed mainly the food supplied by their common dining room. The Yemenites cultivated small tracts of land to raise food which was served from their common din- ing room. The latter ate about 30 grams of fat per day and large quantities of bread, and their intake of animal protein was lower than that of the Europeans. The nutritional status was lower in the Yemenitesx as was their serum cholesterol and 25 lipoprotein concentrations. The incidence and severity of atherosclerosis of the abdominal aorta were about 3 times higher in the European than in the Yemenites. Cohen and coworkers (62) compared the diet pattern of Yemenite Jews before and after their immigration to Israel in order to ascertain whether or not the greater prevalence of heart disease among the long-time residents of Israel could be attributed to a change in diet. While these people lived in Yemen, the main sources of dietary fat had been mutton and beef fat, and butter. The quantity of sugar used had been negligible. In Israel, the Yemenites consumed far more oil, and derived 20 per cent of carbohydrate calories from sugar. This diet was higher in calories than the for- mer. Yemenites living in Israel a longer time had more heart disease than recent immigrants. Keys et al. (63) studied Japanese men in Japan; in Hawaii; and in Los Angeles. Fats contributed 12 per cent, 29 per cent, and 32 per cent of total calories’respectively. The men had cholesterol concentrations of 160, 223, and 253 mg. per 100 ml. of serum. In Japan, heart disease was rare. In Los Angeles, heart disease was as prevalent in the Nisei as in the local Caucasians. In Hawaii, the occurence of heart disease was intermediate. Bronte-Stewart correlated (64) dietary fat and 8.313.014 , I. l. J. _.. .7 r.“ r' 26 cholesterol concentrations in Bantu natives and both colored and white men in Capetown. The Eur0peans ingested 15 gm. of fat from vegetable sources and 83 gm. from animal sources per day. The colored had 20 gm. of vegetable fat and 60 gm. of animal fat. The Bantu consumed 10 gm. of vegetable fat and 40 gm. of animal fat per day. Serum.cholesterol concentra- tions in alpha-lip0protein was 40 mg. per cent for each group. In the beta-lipoprotein, cholesterol concentrations averaged 194.5, 160.5 and 122.3 mg. per cent in the Eur0peans, coloreds and Bantus, respectively. Percentage of cholesterol in the beta-lipOprotein fraction was 82.6 for the EurOpeans, 76.8 for the coloreds, and 71.9 for the Bantus. Under certain conditions diets low in cholesterol and fat effect decreases in the concentration of serum choles- terol. Morrison reported (65) a long-term study of 100 patients with proven coronary atherosclerosis. Fifty of the patients were on self-selected diets, while the remain- der were given a low-fat, low-cholesterol diet containing a total daily intake of 25 gm. of fat. .Animal fats were eaten in.minimum quantities. After 12 years, all of the con- trol group had died but 19 of the experimental group survived. aAt times the degree of dietary restriction necessary to achieve the hypocholesterolemic effect in patients may preclude 27 the use of the regimen. Very low-fat diets may be distress- ing and cause the body to manufacture cholesterol or other lipids from carbohydrate or protein (60). Hatch and others (66) observed this metabolic synthesis in subjects who had been given only 3 grams of fat per day with no cholesterol added. Serum lipid and lipoprotein concentrations did not decline.in.the subjects. Degree gf saturation 2f fats. - Bronte—Stewart has re- viewed (67) the evidence that animal fats, (butter, beef tallow and muscle, lard and eggs) lead to a prompt rise in serum lipid concentrations and vegetable oils, (corn, pea- nut, safflower and olive) lower the serum cholesterol con- centrations. Steiner et a1. (68) placed norma1,atherosclero- tic,and hyperlipemic patients on a diet containing 65-120 grams of safflower oil in addition to carbohydrate and protein adjusted to meet caloric requirements. These pa- tients showed a 21 per cent decrease in serum cholesterol and 5-7 per cent decrease in beta-lipoprotein. The same patients receiving coconut oil in the same amounts showed rapid rises in serum lipids. Page and coworkers (69) found the lowest concentrations of serum lipids in 'pure' vege- tarians. even though 35 per cent of their calories came from fat. 28 Patients given diets containing approximately 100 grams per day of olive oil, usually ingested fewer other lipids. Cholesterol concentration declined 26 per cent and lipopro- teins decreased (70). Boyle and others studied (71) 4 hyper- lipemic patients, all of whom had elevated serum cholesterol with the major portion of the elevations in the very low-den- Sity lipoprotein fractions. Serum lipids decreased strikingly in 2 patients and increased in the other 2. No explanation of this condition was given by Boyle. Beveridge gave (72) diets containing 20 and 60 per cent of calories as corn oil to subjects who had been on a basal fat-free diet. Later, the same subjects were placed on a diet in which butterfat supplied 58.5 per cent of the calories. Plasma lipids increased 28.2 per cent over concentrations during the period of the corn oil diets. Dayton placed (73) elderly institutionalized men on diets containing 38 per cent linoleic acid and 300 mg. of cholesterol per day. The men showed significant drops in serum cholesterol, while men on isocaloric diets with 12 per cent linoleic acid and 750 mg. cholesterol per day had higher cholesterol values. Keys and coworkers investigated (74) men who were in calorie balance. The diets were the same except for the fat. One hundred grams of corn, sunflower seed or fish oils, or 29 butter fatm given. The fat constituted 39.8 per cent of the calories in the diets. Serum cholesterol concentrations rose to 216.9 mg. per cent during the butter fat diet. Corn oil, fish oil and sunflower seed oil depressed serum choles- terol concentrations 52, 39.8 and 35.8 per cent, respectively. Linoleic acid (or arachidonic acid) seems necessary to promote the excretion of cholesterol, but this also depends on the other fatty acids consumed at the same time (75). Keys, Anderson and Grande (76) found no support for the claim that a daily supplement of 4-5 grams of arachidonic acid would depress serum cholesterol. .Bfifiinisgignwgi.Qalggigs. - The effect of calories is less well-defined than the influence of dietary fat on serum lipoprotein concentrations. In an individual in a steady state (not gaining or losing weight) excess calories pg;.§e do not increase serum lipoprotein, according to Ahrens (77). Neither did absolute amounts of carbohydrate, fat, protein and cholesterol have any demonstrable relation to the con- centration of total serum cholesterol. However, the per cent of calories supplied by each was related to the serum cholesterol concentration. 'Walker and others (78) associated positive caloric balance on a low lipid intake (15 grams of fat and 50 mg. 30 of cholesterol) with a significant increase in serum cho- lesterol, and a two-to threefold increase in serum lipo- protein. Since elevated serum cholesterol concentrations contributed to the causation of atherosclerosis, these au- thors proposed that weight reduction was a proper treatment or prevention for the disease. Gofman reported (79) caloric restriction and consequent weight reduction lowered concentrations of low-density lipo- protein. Jolliffe and others (80) found that cholesterol concentrations fell in patients during reducing diets which were low in saturated fats and high in protein from non-' ruminant animal sources and low-fat dairy products. Reduc- ing total caloric intake and reducing the amount of fat in the diet, or simply controlling the amount and type of fat without reducing caloric intake reduces cholesterol in the b1ood, according to the American Heart Association (81). In a study of full-blooded Navajo Indiana, Kositchek and others (82) found a low incidence of coronary artery disease, and a low ratio of alpha-lipoprotein to beta- 1ipoprotein. The diet of these Indians was relatively meager in calories, but relatively high in fats and carbo- hydrates. The findings of Walker (83) in patients on re- ducing diets do not entirely agree. The patients lost an average of 21.7 pounds on a diet in which 73 per cent of i. . it (£14.51... 7 31 total caloric intake consisted of animal fat. In this case, beta-lipoprotein rose, apparently independent of weight change. These data supported the concept that body lean- ness and restriction of dietary fat were both important in preventing atherosclerosis. 5mm: and 51231151 2: melanin. - Wright reviewed (84) human studies in 22 countries. A better relationship was found between the incidence of heart disease and the protein intake than between the prevalence of heart disease and the fat intake. High protein intake resulted in a lower amount of heart disease. On the other hand, British workers stated serum.1ipid concentrations were not influenced by either high or low protein diets (85). Keys et a1. placed (74) two groups on isocaloric diets containing 83 and 130 grams of protein daily. The low pro- tein group received 8.6 per cent of calories from protein; and the high protein group, 17.7 per cent of calories from protein. There wasrx>significant difference in cholesterol concentrations. Connor also found (86) that dietary levels of protein (20 to 143 grams per day) did not influence serum lipid concentrations. in serious protein deficiency may cause a decrease in serum cholesterol. In kwashiorkor, Schendel and Hansen (87) r—1- 32 found exceptionally low concentrations of blood cholesterol as well as a decrease in alpha- and beta-lipoproteins. Sebrell explained (88) the lowering of blood cholesterol by protein restriction. He proposed it could be explained on the basis that a protein deficiency limits the formation of beta-lipoprotein and thereby depresses cholesterol concen- trations, since this moiety is the vehicle for the trans- port of cholesterol. Intakes of vitamins. - The importance of niacin in pro- ducing serum cholesterol changes has been demonstrated. Parsons and Flinn (89) found that nicotinic acid reduced the ratio of alpha-lipoprotein to beta-lipoprotein, but was in- effective in reducing serum cholesterol concentrations. Goldner and Vallan (90) obtained marked and sustained lower- ing of serum cholesterol in humans by medication with a com- bination of niacin and pyridoxine. Oral administration of 3-6 grams of nicotinic acid daily reduced serum cholesterol (91), but this therapy often had adverse effects such as the jaundice reported by Rivin (92). Altschul and Hoffer (93) administered 1 gram of nicotinic acid (buffered with sodium carbonate) 3 times daily for 2 weeks. Serum cholesterol concentrations fell from approxi- mately 200 to about 160 mg. per cent. Minimal. 4" 33 Simonson and Keys (94) have reviewed and Summarized Russian literature on the effects of vitamins on serum lipids and lipoproteins in animals and humans. It appears the studies were done without controls. Vitamin.A was not effective in the prevention and treatment of atherosclero- sis;and in fact, it sometimes produced hypercholesterolemia by eliminating cholesterol from brain and liver. Nicotinic acid was of clinical value. Patients with serum cholesterol concentrations over 250 mg. per cent showed significantly lower concentrations after treatment with ascorbic acid. Administration of 1000 mg. of ascorbic acid for 10 days produced a decrease in beta- and an increase in alpha- lipoproteins in 18 of 35 subjects. Vitamin D (calciferol) increased cholesterol concentrations. Dietary Studies of W 23.9.2.1; WWWW Many factors enter into the aging process. The poor diets of elderly people may accelerate it. Poor teeth and inadequate cooking facilities often cause low consumptions of vitamins and minerals and high intakes of carbohydrates. The latter may lead to weight increase and deterioration of tissues (95). Davidson and coworkers (96) found that the more socially . :pr? .. 34 isolated individuals ate less varied diets and in general, were more above the desirable weight than the more gregarious. The ages of the 104 subjects studied by this group ranged from 50 to 97 years. All were apparently in good physical health. Thirty-nine of the subjects received less than 15 per cent of the total calories from protein. Only those whose teeth or dentures were rated 'good' had high protein intakes (1.5 gm. per kg. of body weight). Some therapeutic diets, such as low sodium, are anorexo- genic. Malnutrition may result. Some diseases, such as Parkinson's disease, make it difficult 1hr the patient to feed himself. Gastrointestinal disturbances following meals may discourage eating. Personality changes due to atheros- clerosis of the brain may affect the appetite (97). WW Sfimm Davidson et al. compared (96) the subjects' dietary in- takes of various vitamins with the National Research Council allowances; 19 per cent of the patients had less than recom- mended intakes of Vitamin A; 40 per cent of thiamin, 67 per cent of riboflavin: 27 per cent of ascorbic acid, and 40 per cent of iron. Ohlson and associates found (98) women aged 52 to 74 years average 300 to 500 calories less per day than younger r .93 «I Q .4. D o I I u- \ r a v‘ \ \ \ " n l l ' \ I _ 1‘ I- \ l e ‘L a \ 35 women. Lyons and Trulson (99) observed that almost half the women studied had caloric intakes well below the recommended allowances, even though 59 per cent of them were 10 per cent or more overweight. In balance studies on women, Ohlson and the others (100) found calcium, phosphorus and nitrogen losses were in excess of intakes when the women consumed less than 1800 calories per day. Batchelder found (101) 58 per cent of the women studied were in negative nitrogen balance whenever caloric intakes fell below 1500 calories per day. From 1947 through 1958, a coordinated research program involving the State Agricultural Experiment Stations of the United States and the Institute of Home Economics of the United States Department of Agriculture evaluated the nutri- tional status of persons of all ages and various economic groups (102). The percentage of calories from protein was practically constant for women aged 20 to past 70, 13.8 per cent. Protein intakes dropped significantly after age 70._ Women past 70 showed dietary averages of 500 mg. of calcium, and low'iron intakes. Some of these investigators felt there was no objective evidence these intakes were low. Estimated allowances for calcium and iron are based chiefly on balance and retention studies, but these are affected strikingly by the level of food calcium and iron to which the subjects are accustomed. Minnelli), . 1,13%}: 114‘ 36 In Lansing, Michigan, Kelley and coworkers studied (103) 54 white and 63 Negro women aged 40 to 90years. Less than 5 per cent reported food intakes providing 80 per cent or more of recommended allowances of calories, protein, calcium, iron, Vitamin A, thiamin, riboflavin and ascorbic acid. Chinn noted (104) in 20 of 500 peOple studied, over- nutrition proved a drawback to rehabilitation after illness or injury. These patients were placed under dietary restrictions but did not adjust easily due to long-established patterns of eating. Ohlson and others also reported (97) obese women re- stricted total calories, but tended to eat sweets to satisfy the appetite, rather than a mixture 0 f proteins, cereals, fruits and vegetables. Usually, this pattern of eating did not bring about weight reduction. Ohlson and associates studied (100) older women, and found that those who maintained vigor for more than 70 years earlier in life. During the late sixties and early seventies, the women reduced food intakes, but kept nutritional equili- brium in their diets. These workers felt that changes in food intakes in old age paralleled loss of vitality. EXPERIMENTAL PROCEDURE Dietary records were obtained from 12 elderly women. Nutrient intakes were calculated and compared with recom- mended allowances. Nutritional status was appraised by physical and bio- chemical examinations, including determination of the con- centrations of serum cholesterol, and relative amounts of serum proteins and lipoproteins. Subjects and Environment The subjects were 12 ambulatory women who lived in a cooperative home in Metropolitan Detroit. A thirteenth resident was deleted from the study because of her inability and/or unwillingness to cooperate with the experimenter. Table 1 lists vital statistics for the 12 participants. The establishment was incorporated and the residents are purchasing the property. The subjects derived their income from one or more of the following sources: Social Security, Social Aid (Old Age Assistance), contributions of children and private income from property, savings and in- surance. No financial support was given by a church or other group. 37 38 TABLE 1 Vital statistics of the subjects SUBJECT ‘ AGE HBIGHTl WEIGHT yr. cm. kg. 1 58 163 61.8 2 67 156 63.5 3 71 166 77.6 4 74 159 55.5 5 76 155 52.4 6 78 161 63.9 7 81 164 87.2 8 82 157 56.7 9 82 154 66.9 10 84 145 53.2 11 89 155 60.2 12 90 157 64.5 1 . ' ‘Wlth two-inch heels. 39 The home was a converted two-story brick duplex resi- dence, with a living room, dining room, kitchen, two bath- rooms, and basement recreation room and laundry. Sleeping accommodations included single and double bedrooms. The residents did the light cleaning and most of them did their personal laundry (automatic washer and dryer). They assisted with the table setting and kitchen cleanup on a rotational basis. Some occasionally helped with food pre- paration. Most felt, however, release from the responsibility of meal preparation was one of the greatest advantages of liv- ing at the home. A regular employee did the general cleaning once a week and a second prepared and served the meals each day. A substitute cook was hired on the day the regular cook was free. Some of the subjects were quite sedentary, because Of chronic illnesses, or lack of vitality and/or interest. On the other hand, some had remarkable vigor. Subject 1 was active professionally (substitute teaching), and took part in many religious, charitable and social functions in addi- tion to managing the home. Subject 2 did most of the food purchasing, and sometimes prepared simple meals. Subject 5 periodically participated in the activities of a "senior citizen"group, but complained of poor eyesight which kept her from such activities as reading and sewing. Subject 12 1' 40 was virtually blind, but did her share of the work. Subject 6 was very active for her 78 years, and maintained a great deal of interest in her personal appearance. She volun- teered for extra household tasks and for some gardening on the premises. Subjects 10 and 11 sewed for charity. The majority of the subjects shared in making charity cancer pads. This project was operated in the home about one day per week. All were free to come and go at will and many visited families and/or friends one or more times a week. No medical personnel were in residence.) A physician was available for house calls. A visiting nurse came to the home free of charge as often as she was needed. Pro- vision for extensive nursing or convalescent care was im- possible. Most of the food was purchased in the neighborhood market. Occasionally donations of government surplus food or gifts from individuals and organizations supplemented food purchases. A supply of canned foods was usually avail- able because of these contributions. A food freezer per- mitted enhanced utilization of perishable foods. The meals were served 'family style” at a common table at regular times each day. (The subjects ate most of their meals at home, but some regularly ate away from home on 41 Sunday evenings when they visited their families. The same food was served to all, but there was some freedom of choice, as individuals could request particular items of food. Some typical menus are shown in Table 2. No attempt was made to regulate the meal patterns or the individual’s selections during the period of study. Dietary Data Enos Intakes -~\‘ Seven-day dietary records were obtained at three dif- ferent seasons: fall, winter and summer. During the fall and winter periods the author instructed the subjects to estimate intakes in household measures, and to record these immediately after each meal on the provided forms (Appen- dix A). The investigator visited the home each day to follow the progress and to give assistance. The cook sup- plied information about the composition of prepared dishes. Subject 3 was not in residence during these first two re- cording periods. To obtain the records during the summer, the author visited the home at each mealtime for the 7 days, and re- corded the intakes. Before the meals, certain foods were weighed or measured in order to have a standard portion for comparison. Intakes were recorded during the meals by a.“ .Lu “i a 12> ¥ in!“ 42 oommoo oHHoo swsam Houusm omenm osasm ousuuon mHouonamU mowmasoxoflso mmmmbm Mae: oumuuso ooxom Hoeusm omowm umonk oaosz usmwxumomm moopmuom poses: xuom unmom MWZZHQ oommoo mom omxooo umom maflmn unmoe nonsense Mas: Hmosflmo canon mmsmuo omscmu Bmdhxdmmm commoo museum nouusm - osoum snowman ooxoou gflflm “$8 ”OGHOU mmmmbw mommou Hnmuxooo passe Houusm omowm fiaoooonm sononm uo>au mama MNZZHQ Tommou Dames omuoousm mmm emxooo umom Rafi: moxmam Deena ocean vasomommuwloammmonwm Emflhfidmmm summou “spasm omoum monsoon voodoo osasm cusses 0.0010 ommuuou - muooso one fiuuonmmmm mammbm moumoo mxmo 050 Houusm ommum mcsom nouns noovmuom omens: seam omxsm mmZZHQ moumou mam moxooo poem Dance oououusm Maw: mcawmm ocean uflswmwmsww Bmfihfidfimm mmzzpm mmHZH3 HoEEss was Housfi3..aasw How Afldh noose Mendoza N Manda ‘ “PM?! 1322 Dflvnlui‘ 5'14 ._ 43 observing the actual food consumption. Subjects were questioned about snacks, which were usually very light. Information was also obtained concern- ing meals eaten away from home. Ordinarily an average of one meal per subject per week was consumed elsewhere. Dur- ing the summer period, subjects 8 and 1 were on vacation part of the time. Subject 8 was away for 4 days, but took the dietary forms with her and recorded all intakes. Sub- ject 1 was absent for the first 5 days of the summer period. Hewever, she completed her set of records on 5 days im- mediately following the regular period. Although these 2 subjects were unsupervised during 4 and 5 days, respectively, their reports were considered accurate. Both had demon- strated competence and conscientiousness in keeping the pre- vious records, and both were very interested and cooperative throughout. 1221mm The handbook (U.S.D.A. Heme and Garden Bulletin #72) (105) was used to calculate the daily intakes of calories, carbohydrate, protein and fat; saturated and unsaturated fatty acids; calcium and iron; Vitamin A, thiamin, ribo- flavin, niacin and ascorbic acid. Whenever possible, the values in the handbook were used. Since niacin values were a...) Hi u Puffeidfg. 1‘ 44 for the preformed vitamin only, niacin mg. equivalents were calculated from the tryptophan on the assumption that tryp- tophan comprises 1 per cent of the protein and that 60 mg. of protein is equivalent to 1 mg. of niacin (106). Appendix A lists certain assumptions and adjustments which were made. The calories contributed by carbohydrate, protein and fat were determined by multiplying the total grams of these nutrients by 4, 4 and 9, respectively. The percentage of total calories contributed by carbohydrate, fat and protein were calculated. Becgmmgngeg Dietary Allowances Recommended allowances were computed according to pro- cedures outlined by the National Research Council (107). Dietary recommendations for a woman 65 years of age were used for calcium, iron, Vitamin A, thiamin, niacin and ascorbic acid. Daily protein allowances were made on the basis of 1 gram per kilogram of ideal body weight. Ideal body weights for a medium frame were selected from height- weight tables (108). The ratios of actual to ideal body weights were determined. These values are recorded in Table 3. Riboflavin recommendations were computed from the pro- tein allowances with the factor 0.025. For individuals 45 TABLE 3 Computed ideal body weights and recommended dietary allowances RATIO RECOMMENDED ALLOWANCES IDEAL ACTUALleEAL FOOD RIBO- SUBJECT WEIGHT WEIGHT ENERGY PROTEIN FLAVIN kg. cal. gm. mg. 1 58.2 1.06 1964 58 1.54 2 54.5 1.18 1707 54 1.54 3 60.5 1.28 1614 60 1.93 4 55.9 1.00 1520 56 1.40 5 53.6 0.99 1473 54 1.35 6 57.3 1.12 1548 57 1.59 7 59.1 1.48 1355 59 2.18 8 55.0 1.03 1284 55 1.42 9 53.2 1.26 1252 53 1.67 10 49.5 1.08 1188 50 1.34 11 53.6 1.12 1259 54 1.52 12 55.0 1.17 1284 55 1.61 46 above average in size, the riboflavin allowance was in- creased by adding 0.025 mg. per kilogram above the ideal body weight (107) . Caloric recommendations were determined on the basis of ideal weights according to the formula: Calories = 0.95 (580 + 31.1 W) where W is the ideal body weight in kilograms (109). Since these caloric recommendations were for women 25 years of age, adjustments were made by decreasing the caloric alloWH ance by 3 per cent per decade between the ages 30 and 50; and by 7.5 per cent per decade between 50 and 70. A further decrement of 10 per cent was employed for each decade past the age of 70. The subtraction of an additional 10 per cent for the decade 80 to 90 was done arbitrarily. Although the National Research Council made no provision for this re- duction, the author felt justified because of apparent de- creased activity of these older subjects. Thus, the energy recommendation calculated from the formula was decreased by 13.5 per cent for the individual aged 58; by 21 per cent for the person aged 67; by 31 per cent for subjects aged 71 to 80; and by 41 per cent for those aged 81 to 90. The allowances for riboflavin and energy are listed in Table 3. 47 Physical and Biochemical Examinations All physical and biochemical examinations were performed during the summer. There is no reason to believe the sub— jects varied appreciably during the study. on each of 3 or 4 visits, trained personnel measured height and weight with laboratory equipment. Means were com- puted and are listed in Table 1. The subjects wore indoor clothing with the exception of shoes. The heights varied due to limitations in the ability of the subjects to stand erect. Two inches were added to each height to allow for 2-in. heels used in the height—weight tables. The physician1 who was in regular attendance at the home made the physical examinations. Heart, pulse and blood pressure data were recorded on Personal History Forms provided by the American Medical Association Committee on Aging.2 These detailed forms were adapted for use in this study. Blood for hematological determinations was obtained by 1Dr. Ezra Lipkin, M.D., Detroit, Michigan. 2Committee on Aging, Council on Medical Service, American Medical Association, 535 Nbrth Dearborn Street, Chicago 10, Illinois. ,Eersgn§1.§istgrx”§grm. Dr. Fredrick Swartz, M.D., Lansing, Michigan was chairman of the comp mittee. 48 finger puncture. On two occasions a medical technician1 collected samples for hemoglobin and hematocrit determina— tions. The same technician performed routine red and white blood cell counts. Each hematological test was performed at least twice for each subject. Each subject collected a morning urine specimen. Physi- cal, chemical and microscopic urinalyses were performed at the University Health Center? The physician3 drew approximately 10 m1. of fasting venous blood on 3 days, at approximately 10-day intervals. Two subjects (2 and 5) refused to give more than one sample of blood. The blood was allowed to clot, and the serum was separated by centrifugation. A portion of each serum was re- frigerated for use in paper electrophoresis, and the remain- der was frozen for cholesterol determinations. mm mem lbi We 'n in The cells were separated from the plasma by centrifuging 1 Employee of the Foods and Nutrition Dept., Michigan State University. 2Arranged by Dr. James Feurig, M.D., Medical Health Service Director, Michigan State University. 3Dr. Ezra Lipkin, M.D. 49 the capillary tubes at 15,000 r.p.m. for 5 min.1 The hema- tocrit was read1 in terms of per cent of packed cells in the total blood volume. Hbmoglobin determinations were made by placing 0.02 ml. of blood in 5 ml. of cyanmethemoglobin reagent.2 Optical densities of the solutions were read.3 Absolute amounts of hemogldbin were determined by comparison of the sample with a standard of known concentration. This method is accurate in practice to i_3 per cent variation. WW .Egngraluprggeggggg. ~13 Durrum-type paper electrophore sis cell4 was used. Veronal buffer of pH 8.6 and ionic strength 0.075 was prepared by dissolving 15.40 gm. of sodium diethyl barbituate and 2.76 gm. of diethyl barbituric acid in distilled water to make one liter. Fresh buffer was used for each determination. A.power supply'maintained a constant current of 125 volts. Schleicher and Schuell filter paper 1International.Micro-Hematocrit Centrifuge, Model MB and International Micro-Capillary Reader. 2Hycel Inc., Houston, Texas. 3Bausch and Lomb colorimeter at 540 mu. 4Spinco Model R paper electrophoresis cell and Spinco Duostat power supply. Procedures were described in Spinco Technical Bulletins TB 6043B and TB 6050A, April, 1958. 50 strips 2034 mgl., 3.0 cm. wide and 30.6 cm. long were placed in the cell and moistened with buffer. The cell was sealed and allowed to equilibrate for 30 min. Samples were trans- ferred by micropipette to a stripper and then applied through a slit in the top of the cell. .Lipgprgtein.agal¥§ifi. - A 0.03 ml. serum sample was used on each paper strip. A constant current of 125 volts was supplied for 16 hr. The strips were dried flat in the oven at 110-1200C. for 30 min. and placed in the stain in vertical position for 18-20 hr. Oil Red 0 dye1 in 60 per cent ethanol was prepared according to the method of Jencks, Durrum and Jetton (1955). The stain was used repeatedly and was kept in a covered cylinder in a 370 C. incubator during equilibration and use. After the dyeing was completed, the strips were washed gently in running tap water at room tem- perature for 5 min., drained, blotted and allowed to dry in air. .Eggtgig analysis. - A 0.005 ml. serum sample was used on each paper strip. A constant current was supplied for 9 hr. The strips were dried flat in the oven for 30 min. at 110-1200C., prerinsed in methyl alcohol for 6 min. and 1National Aniline Corporation. 51 stained for 30 min. in alcoholic bromphenol blue, made by dissolving 1 gm. of the solid dye in a liter of methanol. The stain was stable for 3 or 4 determinations. After 3 rinses of 6 min. each in 5 per cent acetic acid, the strips were dried in the oven for 15 min. Exposure to ammonium hydroxide vapors for 15 min. developed the color before scanning. ‘§ganning.;hg strips. - The filter paper strips were scanned with a recording densitometer} The instrument re- corded the Optical density of the dye in the strips in the form of graphs and sawhtooth integrations. Therelative amount of area included in each portion of the curve could be determined by counting the integrations recorded by the instrument. Wises Total and free cholesterol in the blood sera were de- termined by a modified Schoenheimer-Sperry method. Serum samples of 0.2 ml. were pipetted into a 5 m1. volumetric flask which contained about 3 m1. of 1:1 acetone-alcohol. 1Spinco Analytrol Model RB equipped with 500 mu inter- ference filters, and a B-S com. 1‘ .jl‘éjfl 1w (4%.. \S‘ [be 1 52 The mixture was heated, agitated, cooled and made to volume. The protein, denatured by the solvent and heat treatment, was removed by filtration through fluted fat-free filter paper. One and 2 m1. of this filtrate were pipetted directly into 5 ml. centrifuge tubes for the total and free cholesterol determinations&respective1y. For total cholesterol, 50%.KOH was used foreaponification and consequent release of cho- lesterol from its esters. For both total and free choles- terol, 2 ml. of a solution of 0.5 per cent alcoholic digi- tonin were added to the acidified extracts (alcoholic acetic acid). The precipitated digitonide was allowed to floccu- late in a water bath at 60°C. for about 30 min., and was then stored in the dark at room temperature over 2 nights. The tubes containing the cholesterol digitonide were centri- fuged for 30 min. at about 1500 r.p.m. and the supernatant liquid was decanted. The precipitate was washed once with 1:1 acetone-ether, recentrifuged for 15 min., and the super- natant was again decanted. The washing process was repeated twice with absolute ether. The centrifuge tubes were placed in a sand bath in an oven at 110-1150C. for 30 min. Each pre- cipitate was dissolved in 1 m1. of glacial acetic acid while still hot. After cooling, 2 ml. of cold Liebermann—Burchard reagent (1:20 sulfuric acid and acetic anhydride) were added to each tube. After allowing 30 min. for the development ., .7. s . We. . It? 0.P.!!!“ 1"! u i I r. 53 of color, the optical densities of the cholesterol solutions 1 _ were read at 620 mu. Cholesterol concentrations were de- termined by comparison with cholesterol standards. lBeckman DB Spectrophotometer. RESULTS AND DISCUSSION mm ks Daily nutrient intakes for the subjects are reported in Appendix.A. The mean daily consumptions of food energy, protein, fat and carbohydrate are included in Table 4. Mean caloric intakes for the subjects ranged from 957 to 1692 with a mean of 1342 calories per day. Subjects 8 and 10 ate very small amounts, so their caloric intakes were far be- low the N.R.C. recommended allowances (Table 3), but all other subjects were receiving at least three-fourths of their recommended allowances. Lyons and Trulson also found (99) that 85 per cent of the older people were receiving at least three-fourths of the N.R.C. recommended energy allowance. The average amounts of protein, fat and carbohydrate eaten were 48, 55 and l6lgm., respectively: with ranges of 30 to 67 gm., 36 to 73 gm. and 99 to 212 gm., in the same order. Subject 1, the only person under 60 years of age, consumed the highest amount of protein, 67 gm. No subject over 70 years of age had more than 60 gm. of protein daily: subjects 80 years and over had a mean of 44 gm. per day. Batchelder observed (101) a similar downward trend in protein 54 55 TABLE 4 Mean daily intakes of energy, protein, fat and carbohydrate ' SUBJECT FOOD ENE RGYF PROTEIN? FAT CARBOHYDRATE cal. gm. gm. gm. 1 1692 67 61 167 2 1271 57 55 140 3 1376 52 y 60 160 ‘4 , 1251 ' 36 58 160 5 1318 50 62 144 6 1674 58 73 186 7 1570 45 52 207 8 822 30 36 99 9 1509 58 69 176 10 957 35 40 123 11 1421 50 44 212 12 1278 43 56 160 MEAN 1342 48 55 161 *Recommended allowances were adjusted for individuals and are recorded in Table 3. [1 $11M” uval #999... .I‘ 14 m. u 56 consumption over the years, especially in the sixth and seventh decades. The mean daily intakes of fat and the distribution of the component fatty acids are included in Table 5. Total fat consumption ranged from 36 to 69 gm., with mean satu- rated and unsaturated fatty acid values of 22 and 24 gm., representing 40 and 44 per cent of total fat, respectively. In all cases except one (subject 7), the amount of un- saturated fatty acids equalled or slighly exceeded the amount of saturated fatty acids. However, oleic acid is not an effective unsaturated fatty acid, and when it is omitted, the ratio of unsaturatedzsaturated fatty acids is 0.18. :The variations in butter and meat consumption are reflected in total fat and saturated fatty acid data. The fatty acid data are considered incomplete because values for some foods are not reported in the handbook. Figure 3 shows the individual and mean distributions of calories calculated from grams of protein, fat and carbo- ihydrate. The mean percentages of calories from protein, fat and carbohydrate were 15, 37 and 47 per cent, respectively. Subjects 1 and 2 had 18 per cent of total calories from pro— tein. Subject 4 had the lowest amount of calories from pro- tein, 11 per cent, and claimed she ”never had been a meat eater”. No subject over 70 years of age received more than TABLE 5 Mean daily intakes of fat and distribution of SATURATED FATTY ACIDS TOTAL TOTAL PERCENT OF SUBJECT FAT SAT'D TOTAL RAT . gm. gm. ‘% W i E 4 1 61 23 38 2 55 23 42 3 60 19 32 4 58 22 38 5 62 24 39 6 73 29 40 7 52 25 48 8 36 15 42 9 69 26 38 10 40 16 40 11 44 19 43 12 56 22 39 MEAN 56 22 40 *The mean ratio of unsaturated:saturated fatty acids was 58 the component fatty acids UNSATURATED FATTY ACIDS UNSATURATED OLEIC LINOLEIC TOTAL PERCENT OF SATURATED ACID ACID UNSAT'D TOTAL FAT RATIO 9m- 9m- gm- % 22 6 28 46 1.2 20 4 24 44 1.0 24 6 3o 50 1.6 21 5 26 45 1.2 22 4 26 42 1.1 28 s 33 45 1.1 18 3 21 40 0.8 13 2 15 42 1.0 24 4 28 41 1.1 14 2 16 4o .1-0 16 2 18 41 1.0 20 4 24 43 1.1 20 4 24 44 1.1* 0.18 when oleic acid was omitted. 59 SUBJECT 50-59 years 1 60-69 years 2 , 70-79 years 3 4 5 6 80-89 years 7 8 9 10 11 * 90-99 years 12 *' 0 ‘25 SO 75 100 per cent FIGURE 3 Percentages of protein, fat and carbohydrate calories in the diets in each decade of age. 60 15 per cent of total calories from protein. This is in ac— cordance with the date of Davidson et a1. (96), which showed 40 per cent of aging persons obtained less than 15 per cent of total calories from protein. Although the mean intake of protein was 15 per cent of the total calories, it does not follow that protein intakes were equal to the recommended allowances. Some of the total food consumptions were low, notably those of subjects 8 and 10, so protein intakes did not meet the recommendations for the individuals. Subjects 7 and 11 received more than 50 per cent of their calories in the form of carbohydrate. Both of these persons were in poor health. Subject 7 was almost 50 per cent overweight. Ohlson et al. (100) noted this same pre- ference for carbohydrate in women who experienced loss of health in old age. The subjects' fat intakes comprised from 27 to 42 per cent of total calories. Davidson’s subjects (96) received 26-53 per cent of total calories from fat, and animal fat was over half the total. The mean daily intakes of calcium and iron for each sub- ject are included in Table 6. Calcium intakes of 197 to 597 per day (mean 448 mg.) were considerably lower than the re- commended 800 mg. per day. These low amounts of calcium I.-. m. .u.....i._u..,~.,.. .w .fl 3 61 TABLE 6 Mean daily intakes of calcium and iron SUBJECT CALCIUM IRON mg. mg. 1 539 11.3 2 458 8.6 3 435 7.6 4 453 7.0 5 376 8.3 6 482 10.5 7 553 7.7 8 i 197 5.8 9 ‘ 511 10.2 10 307 6.4 11 597 7.9 12 455 7.3 MEAN 448 8 . 3 N.R.C. RECOMMENDED ALIDWANCES 800 12 .0 62 resulted from poor consumption of milk, although it was available in whole, powdered and evaporated forms, and puddings and ice cream often appeared on the menus. Davidson (96) and Morgan (102) reported increased milk consumption with age, but this trend is not reflected in the subjects of the present study. Iron intakes, ranging from 5.8 to 11.3 mg. per day, failed to meet the recommended allowance of 12 mg. Meats and other food sources were eaten, but the small amounts of these foods which were consumed would account partially for the low iron intakes. These values for dietary iron are not in agreement with Lyons and Trulson (99). whose subjects had mean intakes of 10.00.: 2.4 mg. of iron daily. The mean daily intakes of vitamins are listed in Table 7. Of all vitamins, vitamin A compared most favorably with the N.R.C. recommendations. The mean intake was 7018 I.U. per day, with 9 of the 12 subjects receiving the vitamin in excess of 5000 I.U. per day. These intakes were due to the large amount of this nutrient supplied by butter and eggs which were used freely, and by liver which was served once during the fall and winter recording periods. Carrots and broccoli were common vegetables. Other studies report (102, 103) low intakes of Vitamin A for women, but the sub- jects of the present study are well provided with Vitamin A. 63 TABLE 7 Mean daily intakes of vitamin A, thiamin, ribo- flavin, niacin and ascorbic acid PRE- VIT. RIHJ- FORMED 'IOTAL ASCORBIC SUBJECT A . THIAMIN FLAVIN NIACIN NIACIN ACID I.U. mg. mg. mg. mg.eq. mg. 1 13,380 0.87 1.67 14.6 25.8 126 2 6,199 0.66 1.08 10.4 19.9 30 3 3,823 0.78 0.95 9.4 18.1 51 4 3,313 0.68 0.90 5.8 11.8 '75 5 6,641 0.77 1.29 12.1 20.6 36 6 9,097 0.87 1.39 12.3 22.0 96 7 5,584 0.76 1.26 7.9 15.4 48 8 6,201 0.63 0.78 7.1 12.1 51 9 10,607 0.82 1.54 11.3 21.0 69 10 5,873 0.62 0.99 7.5 13.3 43 11 6,639 0.73 1.31 9.0 17.3 60 12 3,663 0.73 0.91 8.2 15.4 58 MEAN 7,018 0.74 1.19 9.6 17.7 63 N.R.C. RECDMMENDED ALLOWANCES 5,000 1.00 * 17.0 70 *The allowances were adjusted for the individual sub- jects and listed in Table 3. . O a O a a n a a n s a 1 . AECEZSL‘ I! 4 L- .1 64 Thiamin intakes were below the recommended 1 mg. per day. The intakes averaged 0.74 mg. and ranged from 0.62 to 0.87 mg. per day. The mean intake of riboflavin was 1.19 mg. and ranged from 0.78 to 1.67 mg. per day. Since the allowances ‘for this vitamin were based on the weight of the subjects, many of themfespecially those who were considerably over- weight, did not recehe the recommended amounts. Kelley et a1.-reported (103) mean daily intakes of 0.93 mg. of thia- min and 1.26 mg. of niacin in aging women of normal weight. Davidson and coworkers found (96) that 40 per cent of his older subjects took less than 1 mg. of thiamin and 67 per cent toOk less than 2 mg. of riboflavin per day. The niacin values listed in the handbook were for the preformed vitamin. At first, dietary niacin intakes appeared low, but when the niacin equivalents were calculated from its precursor tryptoPhan, the values, for the most part, approached the recOmmended allowance. The mean daily intake of niacin equivalents was 17.7 mg. Trypt0phan was considered to be 1 per cent of total protein, so low values for niacin equivalents were associated with low protein intakes (106). Morgan indicated (102) that niacin deficiencies are unlikely in a person who has a balanced diet with high-quality protein. The subjects of the present study who had good protein intakes also had sufficient niacin. 65 .Ascorbic acid intakes ranged from 30 to 126 mg. per day, with only 3 subjects consuming the recommended 70 mg. per day. The relative absence of raw fruits and vegetables in the diets could be responsible for this. Canned juices were provided for breakfast each day, and the subjects who drank these juices had greater intakes of vitamin C than those who omitted them. Raw cabbage was eaten occasionally, but in very small amounts. Apples were available in the fall, but some of the women did not eat them raw. In the summer, cantaloupe and peaches were served. Low intakes of ascorbic acid are often reported. A survey revealed (102) that nearly 40 per cent of women of all ages 30 to past 80 received less than two-thirds the re- commended allowance of ascorbic acid. In most sections of the country, vitamin C intakes were associated with econo- mic levels, because foods rich in this nutrient may be high in price. In fact, the subjects in the present study have given this as the primary reason why they do not consume more fresh fruits and vegetables. Seasonal variations in nutrient intakes are summarized in Table 8. Subject 3 was excluded from all seasonal compu- tations because she was in residence only in the summer. Mean intakes were at least three-fourths of the recommended allowances, except for calcium and iron. Since the N.R.C. TABLE 8 Seasonal mean nutrient intakes NUTRIENT MEAN INTAKES Energy, cal. Protein, gm. Fat, gm. Carbohydrate, gm. Calcium, mg. Iron, mg. Vitamin A, I.U. Thiamin, mg. Riboflavin, mg. Niacin equivalents FALL WINTER SUMMER MEAN 1523 1353 1150 1342 53 47 45 48 60 57 49 55 178 172 134 161 473 470 401 448 9.7 8.2 6.9 8.3 9624 6976 4454 7018 0.76 0.75 0.71 0.74 1.48 1.14 0.95 1.19 mg. 20.6 17.0 15.5 17.7 57 67 64 63 Ascorbic acid, mg. 67 allowances are generous, these subjects probably have suf- ficient of the various food elements to maintain good nutri- tion. Generally, the subjects had their lowest intakes in the summer. The unpleasant heat probably interfered with the women's appetites. The first two recording periods were in Nevember and February, during which times the subjects stayed in the well-heated home. The lower intakes of the summer would appear serious for nutrients which could not be stored from periods of higher intakes. lllthough the menus contained a variety of foods, they did not differ appreciably from season to season. The con- sumption of canned goods was high throughout the year. Most vegetables were cooked, often for a long time in a large amount of water. Percentages of the N.R.C. recommended allowances sup- plied by the intakes of individual subjects are in Appen- dix A. The seasonal and mean distribution of the subjects consuming less than 50 per cent, between 50 and 74 per cent and 100 per cent or over of the recommended allowances are tabulated in Table 9. In fall and winter, about half of the subjects received the recommended amount of food energy. In summer, only 1 68 TABLE 9 Seasonal and mean distribution of subjects con- of mentioned nutrients PERCENTAGE OF RECOMMENDED FOOD ALIOWANCES ENERGY PROTEIN CALCIUM IRON FALL ‘ Less than 50% 0 0 2 0 50-74% 2 4 8 5 75-99% 3 1 0 5 100% or over: 6 6 l 1 WINTER less than 50%. 0 0 5 0 50-74% 1 3 4 7 75-99% 5 5 1 4 100% or over 5 3 1 0 SUMMER Less than 50% 1 1 8 3 50-74% 2 4 1 7 75-99% 7 4 2 1 100% or over 1 2 0 0 MEAN Less than 50%. 0 0 3 1 50-7¢% 2 3 8 7 75-99% 4 4 0 3 100% or over 5 4 0 0 69 suming specified percentages of the recommended allowance RIBO— NIACIN ASOORBIC __i_[rT . A . THIAMIN FLAVIN EQUIVALENTS ACID 1 0 1 0 2 0 5 3 0 2 2 5 2 4 5 8 l 5 7 2 1 0 2 0 0 0 5 3 l 4 2 6 5 5 4 8 0 1 5 3 ~1 1 2 0 3 4 5 7 3 3 3 5 2 3 3 3 0 0 5 2 0 0 0 0 1. 2 6 6 2 4 0 5 4 3 3 9 0 1 6 3 70 subject had the recommended energy intake. Recommended amounts of protein were taken by 6 subjects in the fall, but by 3 and 2 subjects in winter and summer, respectively. The number of subjects receiving 75 per cent or over of the recommended energy and protein was approximately the same in the three seasons. Most of the subjects received less than 50 per cent of the recommended calcium, and less than 75 per cent of the recommended iron with slightly higher intakes in fall and winter than in summer. However, Morgan shared (102) the opinion of many nutritionists that the calcium and iron requirements are much lower than the recommendations. Al- lowances are based chiefly on balance and retention studies, which are affected by the food levels of these minerals to which the subjects are accustomed. If these recommendations are excessive, the intakes of the present subjects may not be unsatisfactory. Vitimin A intakes were good, with 9 of the 11 subjects having mean intakes higher than the recommended allowances. Thiamin and riboflavin intakes were low, with 6 subjects receiving less than 75 per cent of the recommended allowance. Intakes of niacin equivalents were over 75 per cent of the recommended in most cases. Six subjects had ascorbic acid intakes 75 per cent or over the recommended allowance. 71 Seasonal variations in vitamin intake were not great. .According to the means shown in Table 9, the number of subjects receiving less than 75 per cent of the recommended allowances of mentioned nutrients were: energy - 2} protein - 3) calcium - 11: iron - 8: vitamin.A - 2: thiamin - 6; riboflavin - 6: niacin equivalents - 2; and ascorbic acid - 5. In a nationwide survey of the nutritional status of the peoPle of the United States (102) the average women past 70 had intakes of protein, iron, vitamin A and ascorbic acid less than two-thirds of the recommended allowances. In nearly all cases, the percentage of deficiencies increased with age. In the group studied by the present author, the lowest intakes were in calcium, iron, thiamin, riboflavin and ascorbic acid. The present findings correlated with those of Kelley and associates (103), who studied Lansing, Michigan, white and Negro women aged 40 to 90 years. Less than 5 per cent of these women reported food intakes providing 80 per cent or more of recommended allowances of calories, protein, calcium, iron, vitamin A, thiamin, riboflavin and ascorbic acid. In- takes less than 40 per cent of recommended calcium, vitamin A and ascorbic acid were frequently reported. Lyons and Trulson report (99) iron, calcium and vitamin 72 C were most frequently low in women's diets. They made the best showing in protein, vitamin A and niacin. Five subjects (1, 4, 5, 8 and 10) were within 10 per cent of normal weight; four subjects (2, 6, 11 and 12) were between 10 and 20 per cent overweight, and three subjects (3, 7 and 9) were more than 20 per cent overweight (Table 3). Subject 3 experienced much difficulty of movement due to arthritis, and subjects 7 and 9 were quite sedentary. Many of the subjects reported they had weighed more in earlier years. Figure 4 shows that subjects within 10 per cent of nor- mal weight had mean caloric intakes which were 81 per cent of their recommended allowances; the subjects 10-20 per cent overweight had 99 per cent of recommended calories; and the subjects more than 20 per cent overweight had 118 per cent of recommended calories. These data correlate caloric in- takes with overweight. Since none of the subjects were los- ing weight, the caloric recommendations seemed too high for women of these ages who were no more active than these sub- jects. Dissimilar results were noted by workers (102) who found an inverse relation between obesity and caloric intake, with overweight women reporting dietaries with lower mean energy values than normal weight or underweight subjects. This was only partially explained by the attempts at reducing 1!. 1.: 73 m : 100/o normal weight 10-20% overweight more than 20% overweight % of recommended calories FIGURE 4 Percentage of recommended caloric allowances in the mean intakes of subjects who were normal weight or overweight. 74 weight reported by some of the women. In general, the subjects with the best health also had the best dietary intakes. The meals contained a variety of foods, but some subjects avoided certain dishes or ate very small quantities. Food consumption seemed to depend on ac— tivity and interest. Subject 1, the youngest in the group, did not appear to have any food prejudices and had not changed her dietary habits. Subject 6 was 78 years old, but looked much younger, and was more active than many of the women. Subject 9 was rather senile, but was physically well. The dietary intakes of these 3 women were more in agreement with recommended allowances than the intakes of other less healthy subjects. l’hxsisalansiwoc m’ Simmering Table 10 indicates all subjects had normal hearts, ex- cept two (subjects 2 and 4) who suffered from aortic insuf- ficiency. Subject 4 had an irregular pulse. This subject died of heart failure following surgery to repair a broken bone, (July, 1962). The pulses of all other subjects were regular, the rates ranged from 64 to 108 beats per minute. Subject 2 was definitely hypertensive. The diastolic blood pressure was 254, and the systolic blood pressure was 130 mm. Hg. Three other subjects (3, 8 and 12) had diastolic 75 TABLE 10 Heart, pulse and blood pressure data SUBJECT HEART PULSE BLOOD PRESSURE“ Beats/min. mm. Hg 1 Normal Regular 76 120/70 2 Aortic insufficiency Regular 108 254/130 3b Normal Regular 84 200/90 4 .Aortic insufficiency Irregular 92 190/80 5 Normal Regular 190/80 6 Normal Regular 80 140/70 7a Normal Regular 64 130/50 8 Normal Regular 96 230/100 9 Normal Regular 90 190/80 10 Normal Regular 84 160/80 11 Normal Regular 64 170/80 12 Normal Regular 72 214/90 EAtherosclerosis. b Osteoarthritis of knees. 76 blood pressures over 200 mm. Hg. The oldest subject (12) had a diastolic pressure of 214 mm. Hg. and subsequently died of I"heart stroke' in March, 1962.' Subject 8, whose heart was considered normal, had a rapid pulse, 96 beats per minute; and an elevated blood pressure, 230/100 mm. Hg. Subject 7 was suffering from atherosclerosis, but did not‘ exhibit hypertension (blood pressure: 130/50 mm. Hg.). This subject died of pneumonia in October,l961. Subject 6 had been treated for hypertension and had a blood pressure: 140/ 70 mm. Hg. at the time of the study. The lowest blood pres- sure (120/70 mm. Hg.) was found in Subject 1, the youngest. Mean values for hemoglobin, hematocrit, red and white blood cell counts are listed in Table 11. Hemoglobin values ranged from 12.32 to 15.61 gm. per 100 ml. of blOOd and hema- tocrit values ranged from 37.5 to 46.5 per cent of total blood volume, except for subjects 4 and 5. Red cell counts ranged from 3.64 x 106 to 5.52 x 106 cells per mm. of blood except for the same two persons. Subject 4 had hemoglobin 10.72 mg. per cent, hematocrit 32.38 per cent and red cell count 2.76 x 106 cells per mm. Subject 5 had hemoglobin 8.11 mg. per cent, hematocrit 2.76 per cent and cell count 3.31 x 106 cells per mm. Subject 4 was aware she suffered from pernicious anemia, and reported that she routinely re- ceived injections for this condition. Reticulocyte counts 77 TABLE 11 Hemoglobin and hematocrit cgncentrations and red and white blood cell counts RED WHITE HEMO— HEMAP BLOOD BLOOD SUBJECT GLOBIN TOCRIT CELLS CELLS gm/100 m1. %»of blood million per cells per mm. blood volume mm. blood blood 1 13.23 39.62 4.38 6638 2 15.68 49.5 5.52 7050 3 12.48 39.62 4.84 5875 4 10.72 32.38 2.76 5131 5 8.11 27.6 3.31 6538 6 14.56 44.62 5.15 6038 7 13.46 43.75 3.64 5788 8 13.72 43.38 4.67 10000 9 13.87 42.38 4.16 6206 10 15.68 46.50 4.85 8125 11 12.32 37.50 4.34 6300 12 12.78 38.25 4.90 6988 * Mean values. Individual data are in Appendix 2. r. . VHF": ”I .. 78 counts were made for subjects 4 and 5, and these were nor- mal (0.9 per cent of red cells). White cell counts ranged from 5,000 to 10,000; the high- est count being that of Subject 8, who was apparently suffer- ing from some infection. . It is interesting to note that although the iron intakes were considered low; most of the hemoglobin concentrations are considered normal. The values agree with those found in women aged 17 to 86, in whom the mean hemoglobin was 13.2 mg. per cent, with little difference in women of various ages (100). Batchelder (101) reported the comparable hemoglobin concentration (13.2 gm./100,m1.) in older women. The urines were quite normal (Tables 12 and 13). All were acid to litmus and gave negative sugar tests. The pre- sence of albumin in some of the specimens was not usual of persons of these ages. The cloudiness may have been due to the fact that urinalyses Could not be performed until about four hours after specimens were collected. White cells were present in some specimens,, and these were clumped in the urine of subject 3, possibly indicating a pathological condi- tion. The urine of subject 2 contained budding yeastlike forms, thought to be'a fungus. Casts were found in 3 speci- mens, (subjects 2, 7 and 10), and cylindroid forms, which are not pathological, but may be the forerunner of casts, 79 TABLE 12 SB gigglsand chemical examination of the urine SPECIFIC SUBJECT worm copes GRAVE ALBUMIN 1 cloudy yellow 1.004 negative 2 cloudy 1t. amber 1.018 trace 3 cloudy yellow 1.011 faint trace 4 clear yellow 1.011 negative 5 cloudy 1t. amber 1.015 negative 6 cloudy yellow* 1.014 negative 7 cloudy 1t. amber 1.016 ‘negative 8 cloudy yellow 1.012 trace 9 cloudy straw 1.008 trace 10 cloudy 1t. amber 1.021 faint trace ll cloudy amber 1.025 negative 12 s1. cloudy straw 1.011 faint trace All specimens were acid in reaction and showed negative sugar tests. . . A . - a V . L A . . a a. o .. q 0 a C ._. v” _ -h. l-.‘—.—1-r-—:'w.v!'_ vs.- TABLE 13 Microscopic examination of urine specimens WHITE RED EPITHELIAL SUBJECT BLOOD CELLS BLOOD CELLS CELL§__ 1 8-10 - + 21 16-18 - occasional 3 25-302 0-2 + 4 5-10 - occasional 5 5-15 - 2+ 6 20-30 0-1 occasional 7 2-4 - few 8 3-5 - few 9 2-3* - + 10 25-30 - _ 11 75-100 1-2 few 12 2-3 - few \J lOccasional budding yeastlike forms. 2With occasional clumping. 3Mucous composition, not pathological, forerunner of casts. 81 few BACTERIA CASTS CRYSTALS few - _ few occasional many-Ca 0x hyaline + - - few - _ 3+ - - +++ - urates + + occasional - coarsely granulated ++ - - H» — .. 3+ hyaline 0-11 urates, Ca Ox few occasional3 urobilinogen cylindroid quantitative m”. 1.6., I! BE Av. lg 82 appeared in one sample (subject 3).. Electrophoretic patterns of serum proteins were similar to those reported in the literature (44). A typical serum protein curve is shown in Figure 5. (This curve incloses 62 per cent of the area in the albumin, and 38 per cent of the area in the globulin fractions. The mean value of all the determinations was 66 per cent of the total protein in the albumin fraction, and 34 per cent in the glObulins (Table 14). Subject 4 had albumin concentrations more than 10 per cent above the mean, and subject 3 had concentrations more than 10 per cent below the mean. Goldbloom, using paper electro- phoresis, obtained (44) 60 per cent of the protein in the albumin fraction. Five of the subjects in the present study had mean values more than 10 per cent above this reported normal value. Albumin:globulin ratios of 1:1 to 2:1 are considered normal (34). Only ratios less than 1:1 are in- dicative of liver disease or damage, and this clinical sign does not then appear in the subjects of this study. The character of the lipoprotein fraction varied greatly in the subjects. Figure 6 illustrates differences in subjects 1, 4 and 8. The curve for subject 8 corresponds most closely to the normal values. .Although the same size serum sample was used throughout and the same procedure was followed for all electrophoretic determinations, there was a striking \v Enslfilthli raft». ..4 (m r. 83 . Broom .LE moo; Eofimo 5.30.5 830m 0389305020 339$. m mmDUE ZHEqud WZEDmOAO 7V6 N 84 TABLE 14 Mean percents of albumin and globulin*in serum proteins, and albumin:globulin ratios ALBUMIN GLOBULIN §UBJECT ALBUMIN GIDBULIN RAT;9__ ‘% ‘% 1 69.8 30.2 2.3 2 60.6 39.4 1.5 3 53.0 47.0 1.1 4 71.1 28.9 2.5 5 66.0 34.0 1.9 6 70.8 29.2 2.4 7 62.7 37.3 1.7 8 66.6 33.4 2.0 9 67.1 32.9 2.0 10 72.0 28.0 2.6 11 65.2 34.8 1.9 12 65.8 34.2 1.9 MEAN 65.9 34.1 1.9 * Individual data are in Appendix 3. 85 . Epsom . is no .8 mezzo 530.500: 0339303030 E mcofiotm> 5.32 a ) ~ ‘ . o ‘ ' \ . .5 . 4 ~ . b '. I k." ,' \ '- ‘ \ \- s 1 _.. t u x I . .- . s .‘ . ‘ . . . -. I ~ I a I . .-. \ . . . . _‘ . \ \ TABLE 21 Daily and mean nutrient intakes of Subject 4 7-4—.9 L... . . LL -U'.”.Y“ FATTY ACIDS FOOD PRO- TOTAL UNSATURATED CARBO- DATE ENERGY TEIN FAT SAT'D OLEIC LINDLEIC HYDRATE cal. gn. gm. gn. gn. gn. gm. 11-1 1145 27 44 16 15 2 177 11-2 788 26 40 17 12 4 93 11-3 2357 51 85 36 32 1 350 11-4 1658 59 82 31 32 5 184 11-5 976 36 41 16 11 1 123 11-6 1763 54 79 30 27 9 197 11-7 1158 25 44 17 15 4 179 MEAN 1406 40 58 23 21 4 186 2-14 612 2 29 12 9 1i 81 2-15 1520 31 80 31 36 5 180 2-16 1382 38 58 26 16 2 194 2-17 997 20 41 15 12 6 150 2-18 1542 42 81 30 36 5 173 2-19 1498 35 82 33 27 8 168 2-20 1315 42 75 29 25 9 127 MEAN 1267: 29 64 25 23 5 153 6-30 1041 38 55 23 19 3 211 7-1 965 50 41 13 15 5 109 7-2 1227 46 72 29 28 6 112 7-3 1266 46 60 21 22 5 155 7-4 1332 37 60 20 23 5 179 7-5 749 20 37 9 14 7 94 7-6 976 38 43 15 17 4 119 MEAN 1079 39 53 19 20 5 140 GRAND MEAN 1251 36 58 22 21 5 160 116 267 474 625 357 445 601 345 445 161 183 538 217 250 317 408 296 917 592 602 751 634 261 574 619 453 tommmunmw O ”(d-FOO‘wCD O5 0 \J IFU'IO‘QQUIIh 0 41007410101» 0‘ O O 7.0 RIBO— ASOORBIC VIT. A. THIAMINE FLAVIN NIACIN ACID I.U. mg- m9- m9: mg- 5354 0.60 0.60 5.5. 80 1593 0.41 0.83 2.6 62 5237 1.23 1.20 10.65 53 1813 0.78 .901 11.0 69' 4225 0.59 .89 5.3 43 7555 1.05 1.09 8.6 69 2113 0.40 0.59 3.9 22 4004 0.72 0.89 6.8 57 1368 0.33 0.39 2.9 50 1893 0.52 0.50 4.5 63 2311 0.58 .9 4.4 46 1946 0.45 0.50 4.4 76 1752 0.94 0.70 7.6 31 3211 0.49 0.67 5.5 91 2340 1.25 0.94 7.2 89 1989 0.64 0.67 5.2 64 2714 0.51 1.33 2.6 24 1664 0.82 1.13 5.4 71 3958 0.88 1.28 6.0 - 294 4795 0.93 1.56 7.5 78 2755 0.63 1.08 4.2 71 7343 0.50 0.63 5.6 110 4403 0.49 1.05 5.9 85 3947 0.68 1.15 5.3 105 3313 0.68 0.90 5.8 75 117 - I - \ . | \ o TABLE 22 Daily and mean nutrient intakes of Subject 5 FATTY ACIDS ' . F _ . . - 44 FOOD PRO— TOTAL UNSATURATED CARBO— DATE 4; NE RGY TE IN FAT SAT ' D OLEIC LINOLEIC HYDRATE cal. gm. gm gn. gn. gn. gn. 11—1 1057 34 50 20 19 4 134 11—2 1220 46 63 19 17 7 127 11-3 1561 68 79 33 26 9 156 11-4 1389 56 .73 24 16 Trace 145 11-5 1691 59 75 31 26 1 196 11-6 1456 81 97 41 40 5 133 11-7 1567 60 96 33 29 9 128 MEAN 1421 58 76 29 25 5 146 2-14 1700 53 78 31 28 3 189 2-15 1196 46 41 19 12 2 163 2-16 905 42 37 17 12 Trace 108 2-17 392 28 16 3 2 Trace 36 2-18 2082 63 101 42 46 5 202 2-19 1117 38 42 19 13 4 156 2-20 1752 65 76 34 28 7 166 MEAN 1303 48 56 24 20 3 146 6-30 1378 68 53 25 17 4 166 7-1 595 12 23 5 8 5 90 7-2 1806 77 102 35 40 9 156 743 974 46 57 21 22 4 79 7-4 1580 52 60 28 21 1 196 7-5 942 38 39 15 18 5 105 7-6 1328 42 51 10 24 8 188 MEAN 1229 47 55 20 21 5 140 GRAND MEAN 1313 51 25%, 28 22 4, 144 118 RIBO- ASCORBIC CALCIUM IRON' VIT. A. THIAMINE FLAVIN NIACIN ACID I9. I9. I.U. mg- mg. mg- mg- 172 7.1 5836 0.46 0.52 9.4 42 315 10.1 6132 0.69 1.32 9.8 41 586 12.1 8281 0.70 0.92 13.8 91 411 5.4 2007 0.71 2.90 18.3' 42 457 14.7 50281 0.96 5.07 21.9 44 401 12.8 2235 0.90 1.40 19.3 35 472 12.8 5754 1.24 1.84 11.2 47 402 10.7 11232 0.81 2.0 14.8 49 530 7.4 1637 1.11 0.92 10.7 36 717 5.4 1568 0.54 0.99 9.6 47 331 9.4 32648 0.57 2.90 14.9 54 195 1.9 240 0.34 0.52 8.7 10 216 11.5 2831 0.90 0.98 13.7 31 326 6.4 2426 0.48 0.56 7.5 33 341 9.5 1010 0.72 1.02 15.4 42 379 7.3 6051 0.67 1.06 11.5 36 816 5.6 2120 0.68 1.36 14.7 20 128 1.9 797 0.14 0.21 1.7 4 369 11.7 2867 1.53 1.23 15.4 18 212 6.2 685 1.28 0.67 9.5 10 482 9.6 2175 0.56 1.07 8.2 17 249 6.8 7114 1.16 0.76 8.7 79 180 5.6 2726 0.42 0.44 10.8 15 348 6.8 2641 0.82 0.82 9.9 23 376 8.3 6641 0.77 1.29 12.1 36 n 0 1 a . 3 '- u '- ' \ V . . ' l '\ ‘ 'N I \ 5 . 1 | I J . w . 119 TABLE 23 Daily and nean nutrient intakes of Subject 6 FATTY ACIDS UNSATURATED CARBO— OLEIC LINDLEIC HYDRATE FOOD PRO— TOTAL DATE ENERGY TEIN FAT SAT’D C810 9.0 g‘. g‘. 9.. 9.. 9‘. 11-1 1485 50 82 34 26 6 152 lL—2 1689 54 78 30 27 3 210 11-3 1571 79 84 30 23 3 167 11-4 1477 52 64 24 19 1 212 11-5 2413 102 120 66 48 6 201 11-6 1858 58 80 34 28 3 231 11—7 2152 70 96 34 31 3 252 MEAN 1949 65 72 36 29 4 204 2-14 1821 48 90 38 31 3 219 2-15 1452 54 64 27 19 2 182 2-16 1214 39 46 20 14 Trace 179 2-17 1811 89 91 35 28 1 189 2-18 1847 48 59 24 20 Trace 303 2-19 2130 65 97 39 39 3 240 2-20 1747 60 84 35 30 3 199 MEAN 1717 58 76 31 26 2 216 6-30 1587 71 80 21 31 14 163 7-1 1185 49 61 18 25 7 118 7-2 1428 43 89 25 37 14 127 7-3 1379 47 75 20 30 10. 143 7-4 1736 72 75 24 26 7 194 7-5 1171 41 63 18 24 8 114 7-6 1017 35 56 11 24 9 107 MEAN 1357 50 71 20 29 10 138 GRAND MEAN 1674 58 73 29 28 5 186 120 RIBO— ASCORBIC CALCIUM IRON. VIT. A. THIAMINE FLAVIN NIACIN ACID no mg. lot]. ”9. ‘9. mg. mg. 424 9.9 6866 0.61 0.80 11.2 85 362 9.3 2458 0.64 0.77 9.3 65 795 8.6 3351 0.69 1.43 14.3 71 371 11.8 36724 0.75 3.43 16.0 54 690 18.7 4118 0.96 1.76 20.1 56 408 9.8 9621 0.71 0.95 15.1 71 598 12.7 14678 1.12 1.82 10.9 116 536 11.6 11117 0.80 1.69 14.0 74 556 8.8 2243 0.82 0.94 9.8 182 870 10.0 9960 0.72 2.12 13.1 181 420 12.2 43943 1.23 3.00 16.1 190 819 8.6 8400 0.91 1.75 9.2 158 538 11.3 4509 0.77 0.89 12.3 176 359 13.4 3358 0.92 1.10 15.6 164 408 13.1 2783 0.82 0.79 13.5 50 567 11.1 10744 0.90 1.48 12.8 157 466 7.9 12570 0.70 1.21 13.9 27 350 7.3 2081 1.22 0.92 7.7 70 321 8.7 3870 1.21 1.01 7.6 46 237 9.2 4706 1.43 0.97 9.5 73 494 10.6 2495 0.75 1.30 13.8 32 293 9.1 6822 0.61 0.92 10.2 69 240 8.3 5474 0.45 0.67 7.2 77 343 8.7 5431 0.91 1.00 10.0 56 482 10.4 9097 0.87 1.39 12.3 196 c L o c a o o n . o o . . . . o 4 o o u . . F I C O a v It, DL. 1...» H. II. &\, 121 N l' I I \ \ n .‘ ‘. \ n 1 1 ‘ \ L ---.---.. .-....-. '1 ... '\ I .r "" ._.. . ._.. . ._.. ~Ns .— ... - . ‘ , I 4 . . . . . \‘ 1 . \ D 4 I 1‘. . 1‘ 1 ,. .. \ x n ‘ . . ‘ . TABLE 24 Daily and mean nutrient intakes of Subject 7 mm ACIDS UNSATURATED CARBO— OLEIC LINOLEIC HYDRATB FOOD PRO- TOTAL DATE ENERGY TEIN FAT SAT'D cal. gn. gn. gm. gm. gl. 9!. 11-1 1607 28 31 10 12 2 222 11-2 1823 57 62 25 22 3 224 11-3 1404 31 34 15 10 Trace 210 11-4 1660 51 47 20 16 3 221 11-5 2014 44 63 23 20 3 284 11-6 2233 52 75 30 30 3 298 11-7 1831 43 54 23 19 l 261 MEAN 1789 44 52 21 18 2 246 2-14 1187 51 55 21 20 4 128 2-15 2032 67 87 41 22 4 257 2-16 1641 62 62 30 20 1 219 2-17 1878 69 84 42 28 2 225 2-18 1538 49 58 27 20 1 228 2-19 1914 42 65 28 16 1 288 2-20 1092 28 37 16 11 2 175 MEAN 1612 53 64 39 20 2 220 6-30 1259 40 45 21 18 4 131 7-1 1255 38 24 7 9 3 174 7-2. 1472 37 59 21 24 '4 159 7-3 1422 35 33 12 13 3 209 7-4 1490 43 54 17 21 4 163 7-5 947 26 25 6 8 3 109 7—6 1328 51 46 12 21 5 135 MEAN 1310 39' 41 14 16 4 154 GRAND . MEAN 1570 45 52 25 18 3 207 122 RIBO— ASCORBIC CALCIUM IRON’ VIT. A. THIAMINE FLAVIN NIACIN ACID m. mg. I.U. mg. lug. mg. mg. 248 6.9 7041 0.65 0.74 6.9 49 297 7.9 2538 0.69 0.86 16.5 24 385 5.0 1644 0.50 0.88 5.0 23 828 8.2 3646 0.62 1.39 6.7 15 477 8.6 11557 0.77 1.16 6.2 19 510 2.0 2670 1.25 1.27 10.2 40 590 9.0 3410 0.76 1.22‘ 6.6 56 476 8.2 4644 0.75 1.07 8.3 32 360 8.5 1786 1.25 1.05 8.7 12 1186 9.8 4215 0.92 1.96 6.2 50 978 2.1 38278 1.09 3.82 16.4 123 1515 7.7 8700 0.83 2.09 5.8 18 645 8.9 3850 0.76 1.24 8.2 70 554 9.2 3294 0.96 1.15 9.2 127 372 6.8 2056 0.64 0.74 5.6 64 801 9.0 8883 0.92 1.72 8.6 66 633 6.2 2193 1.18 1.38 4.2 20 420 5.3 1000 0.79 0.95 5.2 67 311 7.4 3845 0.69 1.01 5.6 52 440 5.1 3715 0.44 1.08 5.2 67 444 4.6 2105 0.35 1.01 5.2 7 160 5.9 6376 0.45 0.67 8.0 74 256 6.3 3349 0.38 0.86 13.4 45 381 5.8 3226 0.61 1.00 6.7 47 553 7.7 5584 0.76 1.26 7.9 48 ,. 4 " l 1' .' t .. . , 7. ....... . ..---. r v - . o .‘ o o.‘ a. o . ~ . . , . .\ u . I . l . ‘ 1 - 1 1 . . . . . . . . . . , | '\ l y . , _ . . k , ‘. . . .. . . . \ ' '7 Q ..-. . . 1 ' V \ '-: .‘. .7 . . . ‘ . I Qu ‘ . . . . \ r_ . . . . . . 1 . . . 4. . L. , . ..-. ...... . -....... . . . .u' “MFR?" J TABLE 25 Daily and mean nutrient intake: of Subject 8 FATTY ACIDS FOOD PRO- TOTAL UNSATURATED CARBO- DATE ENERGY TEgN FAT 4§AT'D OLEIC LINOEEIC IUDRATB cal. gm. gm. gn. gm. gn.- gl. 11-1 848 29 39 17 14 1 113 11—2 857 42 40 15 15 4 74 11-3 796 34 33 11 7 l 98 11—4 929 29 39 16 15 l 120 11-5 911 34 51 22 22 3 88 11-6 755 22 28 13 8 Trace 122 11-7 823 34 44 16 17 3 84 MEAN 870 32 4O 16 14 2 102 2-14 1431 38 65 26 25 3 176 2-15 915 30 33 15 10 Trace 128 2-16 787 35 30 12 12 Trace 97 2-17 297 14 8 3 2 2 44 2-18 1070 45 33 13 11 4 132 2-19 866 36 38 19 14 Trace 96 2-20 1420 33 57 18 26 4 203 MEAN 969 33 38 17 14 2 125 6-30 381 13 16 7 6 Trace 50 7-1 588 16 26 ll 9 1 77 7-2 1005 41 63 22, 26 4 76 7-3 777 15 35 9 l7 4 110 7-4 823 49 26 12 10 Trace 84 7-5 462 18 26 12 9 Trace 45 7-6 355 17 13 3 6 2 45 MEAN 627 24 29 11 12 2 69 GRAND MEAN 822 30 36 15 13 2 99 124 RIBO— ASCORBIC CALCIUM IRON VIT. A. THIAMINE FLAVIN NIACIN ACID ”0 m. I.U. m. ”90 mg. mg. 255 5.3 4619 1.14 0.61 7.6 78 114 6.7 1471 0.38 0.82 8.9 58 320 3.8 3330 0.47 0.72 10.1 63 205 8.7 31828 0.46 2.70 10.7 68 236 7.5 3012 0.53 0.70 8.4 29 381 4.4 9315 0.43 0.79 6.4 53 181 6.4 6781 1.17 0.56 5.3 62 251 6.3 8645 0.66 0.98 8.2 59 317 6.5 1391 3.28 0.74 7.8 57 357 4.5 5198 0.33 0.48 9.3 63 198 0.0 34249 0.46 2.60 12.7 126 160 3.1 9690 0.23 0.16 2.7 8 280 8.7 1780 0.56 0.82 10.1 77 145 7.6 1429 0.60 0.56 5.4 61 150 6.0 680 0.35 0.27 8.1 41 230 6.9 7774 0.83 0.95 8.0 62 118 1.7 2899 0.17 0.22 2.0 12 73 3.2 1168‘ 0.61 0.28 4.7 61 160 6.5 2099 0.79 0.61 7.4 54 105 3.5 1288 0.38 0.25 2.9 18 119 7.8 728 0.35 0.56 9.9 2 134 3.5 3834 0.25 0.29_L 3.8 6 71 2.9 3270 0.16 0.75 4.3 67 111 4.1 2184 0.39 0.42 5.0 31 197 5.8 6201 0.63 0.78 7.1 51 . a . . o o c . c c o o o . .. a . o o - \ . . o . . . . . n . . c o . . n . o n . , 4 . . . . 1 . u _ . 9 7,1,4 Al TABLE 26 Daily and mean nutrient intake: of Subject 9 FATTY ACIDS ~25- FOOD PRO— TOTAL UNSATURATED CARBO— DATE ENERGY TEggr FAT SAT'D OLEIC LINDLEIC HYDRATE cal. gn.- gm. gm. gm. gm. gl. 11-1 1802 75 76 32 29 2 225 11-2 1837 64 79 33 25 3 235 1143 1796 85 91 38 26 2 177 11-4 1896 73 76 34 26 1 236 11-5 898 49 35 13 13 3 103 11-6 984 40 53 21 14 1 103 11-7 1688 54 71 26 23 6 218 MEAN 1557 63 69 28 22 2 185 2-14 1309 57 52 23 18 1 165 2—15 1721 59 79 36 24 4 207 2-16 1734 42 72 31 25 3 200 2-17 1122 35 51 23 16 2 145 2-18 2283 65 113 46 48 4 245 2-19 1954 84 91 39 31 6 217 2-20 1865 63 77 32 24 5 257 MEAN 1712 58 77 33 27 4 205 6-30 1191 61 53 17 20 5 133 7-1 1390 56 75 24 28 9 133. 7-2 1478 50 74 25 30 8 165 7-3 1326 47 61 20 25 6 162 7-4 1486 69 65 23 26 4 161 7-5 824 30 34 8 14 4 103 7-6 1108 49 57 13 26 8 113 MEAN 1259 52 60 18 24 6 139 GRAND MEAN 1509 58 69 26 24 4 176 126 RIBO- ASCORBIC CALCIUM IRON VIT . A . THIAMINE FLAVIN NIACIN ACID n9. m9. I.U. mg- mg. mgo m9- 468 13.0 9657 0.95 1.34 17.2 65 679 11.1 5938 0.83 1.32 13.9 84 876 15.6 3523 0.72 1.86 8.9 33 646 14.7 50849 1.03 5.39 20.8 69 345 7.3 1807 0.56 0.83 8.4 37 460 7.2 4458 0.54 0.98 6.9 21 622 10.8 15386 1.06 1.62 8.3 108 585 11.4 13088 0.81 1.91 ' 12.1 59 447 14.0 53318 0.83 4.34 20.3 129 917 9.6 6718 0.84 1.36 10.5 51 449 6.9 1963 0.72 1.51 8.8' 82 362 7.2 2303 0.61 0.72 5.0 60 387 13.0 3429 1.00 1.68 14.1 81 542 15.4 9155 1.14 1.20 17.4 108 856 11.5 . 12529 0.77 1.20 11.1 102 566 11.1 12773 0.84 1.71 12.5 87 497 6.3 11711 0.65 1.21 9.6 18 547‘ 7.9 2540 1.31 1.24 8.0 69 391 9.9 8317 0.91 0.95 10.1 53 325 9.2 4298 1.23 0.99 9.4 80 506 8.6 2400 0.59 1.22 9.6 24 191 7.3 7279 0.57 0.63 8.6 96 226 7.3 5188 0.50 0.73 8.9 80 383 8.1 05961 0.82 0.99 9.2 60 511 10.2 10607 0.82 1.54 11.3 69 0 Q t I o q 4 a ‘ .|.[l.\ . x TABLE 2 7 Daily and mean nutrient intakes of Subject 10 FATTY ACIDS FOOD PRO— TOTAL UNSATURATED CARBO— DATE ENERGY TEIN FAT SAT ' D OLEIC LINOLEIC HYDRATE cal. gm. gm. gm. gm. gm. gm. 11-1 1160 31 47 18 17 4 165 11-2 830 27 39 17 13 3 100 11-3 1001 37 38 16 13 4 133 11-4 920 31 39 14 11 l 120 11-5 1479 41 50 17 18 1 216 11-6 1174 55 48 35 21 3 105 11—7 600 22 17 8 5 Trace 98 MEAN 1023 35 40 18 14 2 134 2-14 1052 35 38 14 15 2 143 2-15 812 23 26 12 10 Trace 127 2-16 877 46 32 15 11 Trace 104 2-17 877 36 24 10 6 Trace 139 2-18 1458 71 107 20 26 3 192 2-19 712 28 21 9 7 l 107 2-20 873 37 36 17 12 Trace 85 MEAN 952 39 41 14 12 l 128 6-30 574 16 19 7 7 1 90 7-1 1029 41 48 17 18 4 115 7-2 1013 29 46 16 18 4 129 7-3 919 32 46 14 18 4 107 7-4 1077 42 51 19 20 3 115 7-5 887 31 4O 20 14 l 101 7-6 776 29 32 12 14 3 98 MEAN 896 31 40 15 16 3 108 GRAND MEAN 3957 35 40 16 14 2 123 128 RIBO— ASCORBIC CALCIUM IRON’ VIT. A. THIAMINE FLAVIN NIACIN ACID m9. m9. I-U- mg. mg. m9. m9. 245 5.6 2775 0.46 0.55 6.8 19 283 4.2 2101 0.32 0.55 3.9 55 223 6.2 1059 0.50 0.49 7.3 15 317 5.7 2955 0.40 0.49 7.6 64 375 0.1 33994 1.84 3.72 15.5 52 239 7.4 1183 0.51 0.76 15.2 55 318 4.9 2879 0.36 1.49 5.0 28 286 7.8 6707 0.77 1.15 8.8 41 314 5.1 529 0.90 0.67 7.2 15 363 3.6 2589 0.30 0.50 4.3 30 315 0.8 47588 0.54 3.97 18.0 83 467 6.6 2330 0.61 0.89 7.1 62 163 7.4 1266 0.52 0.62 7.8 85 237, 5.0 665 0.37 0.39 6.0 31 252 6.0 899 0.42 0.68 8.6 31 302 6.3 7981 0.52 1.10 8.4 48 225 2.5 962 0.28 0.41 2.6 18 542 5.2 1396 0.86 0.76 5.6 34 367 6.3 2841 0.72 0.78 5.2 51 250 5.4 2858 1.08 0.74 6.7 47 402 5.4 1914 0.40 0.90 5.9 9 305 5.6 6025 0.43 0.78 7.4 64 273 4.6 4513 0.33 0.64 4.2 54 332 5.0 2930 0.58 0.72 5.4 40 307 6.4 5873 0.62 0.99 7.5 43 TABLE 28 Daily and mean nutrient intakes of Subject 11 1__“ 15..-.....___._wl" I FATTY ACIDS FOOD PROa TOTAL UNSATURATED CARBO- DATE ENERGY TEIN FAT SAT'D OLEIC LINOLEIC HYDRATE cal. gm. gm. gm. gm. gm. 9!. 11-1 1372 63 41 19 14 l 197 11-2 1516 45 39 17 12 2 251 11-3 1990 56 57 22 20 3 325 11-4 1981 71 84 36 36 4 222 11-5 1652 58 38 17. 11 1 285 11—6 1462 64 52 24 19 2 190 11-7 2066 86 75 29 29 5 273 MEAN 1720 63 56 24 20 3 249 2-14 1685 59 66 30 24 3 224 2-15 1514 65 38 14 11 1.5 237 2-16 1299 36 43 16 17 4 204 2-17 1351 37 49 22 19 2 196 2-18 1133 43 37 16 14 3 163 2-19 1303 37 33 16 ll 1 229 2-20 1641 52 50 20 21 2 261 MEAN 1418 47 45 19 16 2 216 6-30 1185 53 32 13 11 l 182 7-1 1379 47 39 15 14 3 212 7-2 973 25 34 15_ 11 2 149 7-3 1233 35 37 15 14 3 203 7-4 1338 53 39 17 14 1 188 7-5 813 33 20 7 7 1 118 7-6 944 36 26 9 10 2 147 MEAN 1124 40 32 13 12 2 171 GRAND MEAN 1421 50 44 19 16 2 250 130 ASCORBIC CALCIUM IRON VIT..A. THIAMINE FLAVIN NIACIN .ACID n9. n9. 1-0- mg. mg. m9. m9- 818 13.7 52747 0.82 4.70 26.7 118 780 6.5 5258 0.48 1.11 8.4 18 710 9.5 1826 0.92 1.27 9.2 58 978 9.7 3085 0.76 1.84 11.6 33 794 11.2 3417 0.90 1.51 7.9 57 764 9.2 3107 0.64 1.26 10.6 88 803 10.0 2220 0.65 1.50 11.5 65 807 10.0 10237 0.74 1.88 12.3 62 759 9.2 1870 1.27 1.48 9.2 29 910 8.7 13733 0.87 1.42 17.9 135 436 7.5 4471 0.77 1.01 6.2 48 524 5.6 1658 0.73 1.07 4.4 61 412 .4.9 5724 0.40 0.86 5.3 29 679 6.4 4797 0.74 1.12 8.7 100 598 8.7 7801 0.73 1.15 7.5 106 615 7.3 5723 0.79 1.14 8.3 73 493 6.1 11413 ' 0.49 1.13 6.0 33 494 6.6 1466 1.03 1.03 6.1 66 326 5.0 2271 0.63 0.70 4.8 26 317 7.4 3580 0.91 0.90 _ 6.3 81 463 8.4 1753 0.67 1.17 7.8 35 156 6.2 3277 0.44 0.58 8.4 47 334 5.5 3938 - 0.41 0.78 5.7 35 369 6.4 3957 0.65 0.90 6.4 46 597 7.9 6639 0.73 1.31 9.0 60 TABLE 29 Daily and mean nutrient intake. of Subject 12 FATTY ACIDS FOOD PRO- TOTAL UNS ATURATED CARBO- PATE ENERGY TEIN FAT SAT ' D OLEIC LINOLEIC HYDRATE cal. gm. gm. gl. 93. 9!. 9|. 11-1 1842 45 77 32 26 2 263 11-2 1107 37 49 21 14 2 145 11—3 1241 47 58 18 15 7 148 11—4 1861 67 67 28 22 7 233 11-5 810 25 32 15 10 2 115 11—6 1971 63 97 39 39 6 201 11-7 1195 37 40 15 14 4 185 MEAN 1433 46 60 24 20 4 184 2-14 1479 48 60 23 19 9 201 2-15 1151 26 52 18 18 7 156 2-16 1320 41 55 21 17 5 182 2—17 1210 33 32 13 11 Trace 211 2-18 1402 41 65 30 19 4 186 2-19 1382 36 68 27 26 2 170 2-20 1507 41 72 29 27 4 184 MEAN 1351 38 58 23 20 4 184 6-30 1499 67 70 22 25 11 166 7-1 1239 58 49 16 18 7 148 7-2 540 19 26 10 10 2 62 7-3 1208 48 72 24 28 7 106 7-4 1352' 59 63 25 26 3 134 7-5 835 29 32 9 14 3 104 7-6 680 34 32 11 13 3 72 MEAN 1050 45 49 19 19 5 113 GRAND MEAN 1278 43 56 22 20 4 160 132 RIBO- ASCORBIC CALCIUM IRON VIT. A. mums FLAVIN NIACIN ACID mg- mg- I-U. mg- m9° m9- m9- 572 8.2 2907 0.95 1.00 7.8 47 606 5.8 4028 0.60 0.84 8.9 35 293 7.3 3194 0.52 0.87 12.2 69 466 13.7 2540 0.87 1.28 12.9 89 341 4.7 1328 0.46 0.53 4.1 21 204 11.7 2400 0.95 0.90 14.7 95 433 6.0 1676 0.49 0.79 5.2 13 416 8.2 2296 0.69 0.89 9.4 53 236 9.8 7536 1.58 0.60 13.2 115 352 5.5 1597- 0.47 0.55 5.4 76 666 8.2 10390 0.48 0.86 7.8 42 344 6.9 1983 0.52 0.50 5.7 70 994 6.0 2394 0.74 1.33 4.9 41 302 7.1 2832 0.80 0.78 8.2 30 265 7.7 1712 1.26 0.81 9.9 84 452 7.3 4063 0.84 0.78 7.9 65 780 6.4 12174 0.59 1.43 11.0 29 735 8.0 1849 1.11 1.38 6.2 90 332 2.9 868 0.32 0.53 2.3 4 526 8.0 4918 1.16 1.29 7.7 74 650 7.5 2673 0.60 1.41 7.6 28 139 7.1 6872 0.52 0.57 10.4 106 327 4.8 3048 0.31 0.76 5.0 62 498 6.4 4629 0.66 1.05 7.2 56 455 7.3 3662 0.73 0.87 8.3 58 133 vvumrvw TABLE 30 Weekly and mean percentages of the recemnended FOOD SUBJECT SEASON ENERGY PROTEIN % % 1 Fall 120 143 'Winter 58 86 Summer 86 119 Mean 88 116 2 Fall 72 106 Winter 84 109 Summer 67 100 Mean 74 106 3 Fall Winter Summer 85 87 Mean 85 87 4 Fall 92 71 ‘Winterf 83 52 Summer 71 70 Mean 82 64 5 Fall 96 107 Winter 88 89 Summer 83 80 Mean 89 93 6 Fall 126 114 Winter 111 102 Summer 88 88 Mean 108 102 CALCIUM ‘% 70 47 85 67 56 73 43 57 54 54 56 37 77 57 50 47 44 47 67 71 43 60 IRON %» 127 63 93 94 77 79 58 72 63 63 70 56 50 58 89 61 57 69 97 92 47 88 134 allowances in the nutrient intakes RIBO- ASCORBIC VIT. A. THIAMIN FLAVIN NIACIN ACID '% ‘% ‘% %» ‘% 462 100 166 125 168 119 67 65 49 70 222 95 95 84 300 , 268 87 108 96 180 216 60 84 7o 38 96 62 62 64 so 60 76 59 49 40 124 66 68 61 43 76 78 49 55 73 76 78 49 55 73 80 72 64 4o 81 4o 64 48 3o 91 79 68 82 31 150 66 68 64 34 107 225 81 148 87 70 121 67 81 69 51 53 82 ‘ 61 58 33 133 77 96 71 51 222 80 106 82 106 215 90 93 75 224 109 91 63 59 80 182 87 87 72 137 fl w 135 TABLE 30 Weekly and mean percentages 0f the recommended FOOD SUBJECT SEASON ENERGY PROTE;N grown IRON % % ‘% % 7 Fall 132 74 60 ‘68 ‘Winter 119 90 100 75 5?. Summer 97 66 48 48 { Mean 116 76 69 64 7” 8 Fall 68 58 31 52 ‘Winter 75 6O 29 58 Summer 49 44 14 34 Mean 64 54 25 48 9 Fall 124 119 73 95 ‘Winter 137 109 71 92 Summer 100 98 48 68 Mean 120 109 64 85 10 Fall 86 7O 36 65 Winter 80 78 38 52 Summer 75 62 42 42 Mean 80 70 38 53 11 Fall 137 117 101 83 Winter 113 87 77 61 Summer 89 74 46 53 Mean 113 92 74 66 12 Fall 112 84 52 68 Winter 105 69 56 61 Summer 82 82 62 53 Mean 100 78 57 61 136 allowance: in the nutrient intakes--continued RIBO— ASCORBIC ‘le. A. THIAMIN FLAVIN NIACIN ACID % % % % % 93 75 49 49 46 178 92 79 50 94 64 61 46 39 67 112 76 58 46 ' 68 173 66 69 48 83 155 83 67 v 47 88 44 39 30 29 44 124 63 55 42 71 262 81 114 71 84 255 84 102 74 124 119 82 59 54 86 212 82 92 66 98 134 77 86 52 58 160 52 82 49 68 59 58 54 32 57 117 62 74 44 61 205 74 124 72 88 114 79 75 49 104 79 . 65 59 38 66 133 73 86 53 86 46 68 55 55 76 81 84 48 46 93 92 66 65 42 80 73 73 56 48 83 [I117 .ulh‘ll' APPENDIX B Laboratory data 137 TABLE 31 Hemoglobin, hematocrit, red and white blood cells counts RED WHITE HEMO— HEMATO— CELL CELL '§UBJECT DATE SAMPLE GLOBIN CRIT COUNT COUNT gm/lOO %»of million cells/mm. ml. blood blood per mm. blood volume blood 1 7-8 1 13.23 39.0 2 13.23 39.5 7-15 1 40.0 4.33 6950 6700 2 4.43 6850 6050 2 7-8 1 15.46 49.0 5.06 5.97 7450 6650 2 15.89 50.0 2 12.87 5.67 5.14 5700 5150 7-15 1 41.5 5.10 2 41.0 4.98 4 7-8 1 11.13 31.5 5650 5950 2 10.01 7-15 1 10.36 33.5 2.76 4750 4300 2 11.39 33.0 2.77 4400 4400 5 7-8 1 8.39 26.0 2.43 7400 7950 2 26.5 3.30 7-15 1 8.11 29.0 3.75 5200 5600 2 18220 3 8.03 6 7-8 1 14.85 44.0 3.51 5950 5950 2 44.0 3.54 6400 5850 7-15 1 14.26 45.0 4.70 2 45.5 5.60 ‘A‘Ill.l".l..ll. It‘ll). 138 TABLE 31 Hemoglobin, hematocrit, red and white blood cells counts--continued RED WHITE HEMO— HEMATO- CELL CELL SUBJECT DATE SAMPLE GLOBIN GRIT COUNT COUNT gm/100 ‘% of million cells/mm. ml. blood blood per mm. blood volume blood 7 7-8 1 13.59 44.0 3.62 6350 5700 2 13.32 43.5 3.65 5900 5200 8 7-8 1 14.25 45.0 2 14.15 45.0 '7-15 1 13.77 41.5 4.63 9750 9550 2 12.72 42.0 4.70 10350 9 7-8 1 13.32 42.0 7100 6950 2 41.5 7-15 1 14.42 43.0 4.74 5650 5600 2 3.59 5150 10 7-8 1 15.68 46.0 4.94 7150 8500 8450 8800 2 47.0 4.76 7900 8150 11 7-8 1 12.18 36.0 6200 6400 2 38.0 7-15 1 12.46 38.0 4.42 6200 6400 2 38.0 4.27 ‘ 12 7-8 1 12.95 37.0 4.45 7050 7550 2 3.52 6500 6850 7-15 1 12.62 39.5 5.80 139 TABLE 32 Distribution of fractions of serum proteins PERCENT OF TOTAL PROTEIN I , . $12. 'J. 34:“. .. 1 PE; "in GLOBULINS - SUBJECT DATE ALBUMIN ALPHAl ALPHA2 BETA GAMMA ‘% ‘% ‘% '% 7-19 70.2 2.9 5.8 9.6 11.5 7-26 66.7 2.7 6.3 9.9 14.4 7-26 73.6 2.8 5.7 7.5 1004 2 7-19 59.1 3.4 8.0 14.8 14.8 3 7-19 54.9 1.2 3.7 12.2 28.0 7-19 49.5 3.3 6.6 11.0 29.7 7-26 55.7 3.1 8.2 11.3 21.6 7-26 51.8 3.6 7.1 9.8 27.7 7-19 71.7 3.3 5.4 5.4 1 7-26 76.9 3.8 5.1 501 5 7-19 66.0 4.0 4.0 12.0 14. 7-19 74.4 7.5 7-26 66.4 11.8 7-26 53.6 12.0 140 TABLE 32 Distribution of fractions of serum proteins-- continued PERCENT OF TOTAL PROTEIN GIDBULINS ~ SUBJECT DATE ALBUMIN ALPHAl ALPHA2 BETA GAMMA ‘% ‘% ‘% %1 ‘% 7-19 65.2 3.4 10.1 .10.1 11.2 7-26 65.2 4.3 8.7 7.8 13.9 7-26 68.2 2.7 7.3 7.3 1405 9 7-19 71.2 2.5 7.5 5.0 13.8 7-19 62.2 3.1 8.2 9.2 17.3 ‘ 7-26 67.6 1.8 9.0 9.9 11.7 7-26 59.1 3.5 10.4 11.3 15.7 10 7-19 73.3 4.2 7.5 8.3 6. 7-19 71.3 2.6 6.1 13.9 60 7-26 69.9 3.2 7.5 11.8 7. 7-26 73.5 2.9 7.4 10.3 5. 7‘26 66.2 3 .4 8.8 10.8 10.8 7-19 63.9 4.2 6.9 9.7 15.3 7-26 62.5 1.7 8.3 10.8 16.7 7-26 66.4 3.2 604 7.2 16.8 141 TABLE 33 Distribution of fractions of serum lipoproteins DATE OF LIPOPROTEINS SERUM PERCENT OF TOTAL SUBJECT COLLECTION ALPHA BETA % % 8-8 707 92.3 8-8 9.1 90.9 8-8 7.6 92.4 2 3 7-19 13.1 86.9 7-26 24.8 75.2 7-26 16.0 84.0 8-8 25.7 74.3 4 7-19 12.9 87.1 8-8 15.2 84.8 8—8 18.4 81.6 5 7-19 9.3 90.7 6 7-19 6.7 93.3 7-19 8.9 91.1 8-8 9.4 90.6 8-8 13.2 86.8 8-8 11.8 88.2 7 8-8 7.4 92.6 8-8 5.4 94.6 8-8 6.1 93.9 142 TABLE 33 Distribution of fractions of serum lipoproteins ' ;--continued DATE OF LIPOPROTEINS SERUM PERCENT OF TOTAL SUBJECT COLLECTION ALPHA BETA ‘% ‘% 8 7-19 28.5 71.5 8-8 34.9 65.1 8-8 31.0 69.0 9 7-19 15.1 84.9 8—8 7.6 92.4 828 6.1 93.9 8-8 16.8 83.2 10 7-26 19.1 80.9 8-8 18.9 81.1 8-8 14.5 85.5 8—8 15.4 84.6 11 7-26 15.6 84.4 8-8 9.4 90.6 8—8 8.1 91.9 8-8 2.8 97.2 8-8 8.7 91.3 12 7-19 14.7 85.3 7-26 27.4 72.6 7-26 21.2 78.8 8-8 15.2 84.8 8-8 14.0 86.0 8-8 11.3 88.7 143 TABLE 34 Total and unesterified serum cholesterol concene trations . 5 DATE OF SERUM CHOLESTEROL SUBJECT COLLECTION 'IOTAL UNESTERIFIED mg.% mg.% 251.2 50.0 282.0 55.5 313.2 62.2 8-8 300.0 52.4 261.8 51.5 2 7—19 268.0 68.1 283.0 64.8 3 7-19 215.0 53.1 244.8 41.5 7-26 259.8 56.4 221.2 59.9 253.2 8-8 193.2 42.2 209.5 44.9 225.0 4 7-19 119.8 25.9 116.2 27.5 7-26 102.8 24.0 100.0 22.2 100.0 111.0 88.0 144 TABLE 34 Total and unesterified serum cholesterol concen- trations--continued DATE OF SERUM CHOLESTEROL SUBJECT COLLECTION TOTAL UNESTERIFIED _ mg.% mg.% 5 7-19 223.0 54.9 209.8 56.4 56.4 6 7-19 177.2 43.2 7-26- 238.0 50.0 219.5 50.6 8-8 198.2 52.4 201.2 44.0 7 7-19 190.0 50.0 223.0 41.5 7~26 158.5 39.0 183.5 39.0 142.0 36.5 8 7-19 .204.8 45.8 221.2 43.1 226.2 7-26 203.0 50.6 190.0 42.4 173.2 45.8 145 TABLE 34 Total and unesterified serum cholesterol concen- trations--continued DATE OF SERUM CHOLESTEROL SUBJECT COLLECTION 'IOTAL UNESTERIFIED mg.% mg.% 9 7-19 195.0 44.0 231.2 44.0 7-26 226.2 55.5 246.2 51.4 8-8 215.0 51.5 190.0 43.1 .10 7-19 143.2 31.5 178.8 30.8 34.0 170.0 38.1 11 7-19 248.0 60.6 250.0 60.6 7-26 225.0 55.5 241.2 61.5 8-8 177.0 38.1 203.0 46.5 12 7-19 162.2 35.6 178.8 30.8 188.2 36.5 156.8 36.5 183.5 '7' . 'i .I A“... V " 19 “CE ”LLB a -r ‘4' 5 4‘ E’sd’ifin: Us; (11% I MITITIIIWWIWI lul 1111111171111 NW ES 3 1193 03082 9 46 6