Mifiifiaéemm Sitate 4 Umw witty ‘-—-—.—._...~. _ This is to certify that the dissertation entitled SENSORY RESPONSE TO FOOD STIMULI. IN THE ELDERLY AND COLLEGE-age SUBJECTS presented by STEVEN ANTHONY WITHERLY has been accepted towards fulfillment of the requirements for PH . D . degree in _EQ_QD__$_C_LENC_E 8 HUMAN NUTRITION Rachel Schemmel Major professor Date 11-12-82 MS U is an Affirmative Action/Equal Opportunity Institution , 0-12771 f MSU RETURNING MATERIALS: Place in book drop to LJBRARJES remove this checkout from w your record. FINES will - be charged if book is returned after the date stamped below. SENSORY RESPONSE TO FOOD STIMULI IN THE ELDERLY AND COLLEGE-AGE SUBJECTS By Steven Anthony Hitherly A DISSERTATION Submitted to Michigan State University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Food Science and Human Nutrition 1982 ABSTRACT SENSORY RESPONSE TO FOOD STIMULI . IN THE ELDERLY AND COLLEGE-AGE SUBJECTS By Steven Anthony Witherly The purpose of this dissertation was twofold. First, to compare the sensory responses of gustation, olfaction and saliva- tion to food stimuli between two divergent age populations (elderly and college-age subjects) and second, to study the effect of zinc and copper supplementation on the restoration of decreased olfactory and salivation function in a group of eight elderly subjects. In Study 1, a comparison of sensory responses to gustation (saltiness and sweetness, both hedonic and intensity) and olfaction (intensity, odor recognition) was studied in 25 healthy elderly subjects (mean age: 83.0; SD::5.0) and college students (mean age: 24.0; 50:17.2). In addition, whole-mouth salivary flow rates were measured in response to sniffing and tasting lemon wedges and lemonade. Utilizing a 17-point, category structured scale, the taste hedonics and intensity responses to 6.0, 8.0, 10.0, 14.0, and 18.0% w/v sucrose in lemonade and 0, 1.0, 2.5, 4.5, and 7.0% NaCl in unsalted tomato juice were studied. Hedonic responses peaked for both groups at the 1.0% w/v NaCl level in tomato juice and the 14.0% w/v sucrose in lemonade. The positively sloped intensity reSponses to saltiness and sweetness were similar, although the elderly demonstrated a narrower range of response than the college students. In the olfactory studies, subjects compared odor recognition of ten food odorants, including the intensity of the first 7 using an unstruc- tured 100mm line scale. Correct identification of the ten food odors averaged 86% and 33.5% for the college and elderly, respectively. Compared to the elderly, college students rated the seven odorants as being significantly more intense (mean: 61.4 vs. 42, t= 4.49, pstOI). In the final study, 2-minute salivary flow rates were measured under various conditions. Resting levels of salivary flow were similar for both groups. However, unlike the college students, salivary flow in the elderly did not increase due to sniffing lemons and increased only moderately in response to tasting lemonade. In Study II, two additional olfactory experiments, utilizing the subjects of Study I, examined whether the elderly would improve their odor recognition scores with practice, and what classes of foods, if any, the elderly might assign to blank sampling cups included with the eight food aromas. As in Study I, the elderly scored below 50% recog- nition for the majority of the eight food odorants, even when a listing of the foods was placed inside the tastebooth for inspection. Elderly subjects recognized the blank sample cups as blank 50% of the time, with no discernible pattern of food classification in their guessing. In contrast, the college-age subjects approached 100% correct identi- fication for the eight food aromas and two blank sampling cups. Finally, in Study III, the incorporation of 50 mg of zinc (as zinc citrate), and 5 mg of capper (as an amino acid complex) in the diets of eight elderly for eight weeks had little effect upon the elderly's perception of aroma intensity, olfactory recognition or salivation to food stimuli as compared to the previous baselines established before supplementation. Some subjects, however, noticed an increase in general appetite, while one subject demonstrated a remarkable increase in overall salivation (2-3 times greater). Analysis of the hair in Study I revealed no significant difference in zinc level between the college (mean: 214.5 :_61 pg/g) and elderly (mean: 188.3 :_47 ug/g); while copper levels were significant at pvuuu=mmocaoc m.me "x .mw-m~ moan .mopos ma Amhaav aa,;z gamma. wows: Ahamfiv mama new me_o3 Lemaav L._=oum ac. apnn_h .....Aommfiv ...e “a ae__z ....eahmfiv ..pm um coxJ .II.Ihmnm~v ..pn um yea: ......Amfimsv ..P. a. emcee: npmmwp a._.fl Hm weapon: Amumfiv camcapma ecu weapon: “may am away :N cocoa: muuannam mcoguz< ".mucmnzum ommP—ou c. axons. agaum.o use Lmqqou\o:.N ea movcaum .e mpneh 10. 11. 12. REFERENCES Harland, 8.F., Johnson, R.D., Blenderman, E.M., Prosky, L. , Vanderveen, J.E., Reed, G.L., Forbes, A.L. and Roberts, H.R. Calcium, phosphorus, iron, iodine, and zinc in the "Total Diet". J. Am. Diet. Assoc., 1977, 16-20, 1977. Harland, 8.F. and Peterson, M. Nutritional status of lacto-ovo vegetarian trappist monks. J. Am. Diet. Assoc.,72, 259-264, 1978. ‘ Holden, J.M., Wolf, W.R. and Mertz, W. Zinc and copper in self-selected diets. J. Am. Diet. Assoc., 75, 23-28, 1979. Hunt, 1.F., Murphy, N.J., Gomez, J. and Smith, J.D. Dietary zinc intake of low-income pregnant women of Mexican descent. J. Am. Clin. Nutr., 32, 1511-1518, 1979. Lyon, T.D.B., Smith, H. and Smith, L.B. Zinc deficiency in the west of Scotland? A dietary intake study. J. Br. Nutrition, 42, 413-416, 1979. Milne, 0.8., Schnakenberg, 0.0., Johnson, H.L. and Kuhl, G.L. Trace mineral intake of enlisted military personnel. J. Am. Diet. Assoc., 76, 41-45, 1980. Tribble, H.M. and Secular, F.I. Zinc metabolism of young college women on self-selected diets. J. Nutr., 52, 209-216, 1954. Walker and Page, L. Nutritive content of college meals. J. Am. Diet. Assoc., 66, 146-152, 1975. Walker, M.A. and Page, L. Nutritive content of college meals. II. Lipids. J. Am. Diet. Assoc., 68, 34-38, 1976. Walker, M.B. and Page, L. Nutritive content of college meals III. Mineral elements. J. Am. Diet Assoc., 70, 260-66, 1977. White, H.S. Inorganic elements in weighed diets of girls and young women. J. Am. Diet. Assoc., 55, 38-43, 1969. White, H.S. Zinc content and the zinc-to-calorie ratio of weighed diets. J. Am. Diet. Assoc., 68, 243-245, 1976. 48 C.1. Nutritional, Biochemical and Pharmacological Roles, Requirements and Toxicities of Zinc and Copper Several excellent and comprehensive reviews exist delineating the functional relevancies of zinc and copper in human, animal, and microorganism nutrition. Prasad (1979b), an early pioneer of zinc inadequacy in human diets, provides a concise review of the trace elements, zinc and copper, from a biochemical and clinical standpoint. A more detailed work by Prasad (1979a) carefully delineates the clinical, pharmacological and biochemical roles of zinc in human nutrition. A summary of the more important aspects of the two references cited above follows. Zinc, a mineral of molecular weight of 65, is a normal constituent of body fluids, enzymes and bone. A 70 kg male contains approximately 1.5-2.0 grams of zinc, concentrated in liver, kidney, bone, retina, prostrate and muscle. Zinc in the plasma is mainly bound to albumin, but other proteins such as ceruloplasmin and a-macroglobulin may bind significant quantities. Of interest is the zinc-binding abilities of several amino acids (histidine, glutamine, threonine, cystine and ly- sine). Zinc absorption, influenced by such factors as body size, phytate content and chelating agents in the diet, varies considerably in man, with approximately 20-30% of the ingestate absorbed. Animal proteins usually provide greater zinc availability than plant proteins (Prasad, 1979). Normal zinc intake in a well-balanced diet approximates 12 to 15 mg/day. 49 50 Urinary, sweat and gastrointestinal zinc losses can be considerable (3-4 mg/day) depending, of course, on climatic or disease conditions. The major function of zinc in biological systems centers around the 70 metalloenzymes known to require zinc as a catalyst or as part of the apoenzyme complex. As such, the metal may be found in several dehydrogenases, aldolases, peptidases, and phosphatases. Deficiency of zinc in man occurs in a wide variety of diseased states or syndromes. A summary of the more important medical conditions is listed in Table 5. Typical syndromes of mild zinc deficiency include growth retard- ation, hypogonadism in males, skin changes, poor appetite, mental lethargy and delayed wound healing. In severely zinc-deficient patients, dermatologic manifestations, diarrhea, alopecia, mental disturbances-and recurrent infections predominate. For comparative purposes, Table 6 lists some biochemical functions and manifestations of zinc and copper with their molecular weight relatives--vanadium (51), chromium (52), manganese (55), iron (55.8), cobalt (60), and nickel (58.7). Copper has received extensive attention from reviewers in the past decade (Evans, 1973; Prasad, 1979; and Mason, 1979). The presence of capper in plant and animal tissue was first recognized almost 150 years ago. The earliest observation was the role of copper in promoting hematOpoesis. Copper is widely distributed in the body, with the highest concen- trations in the liver and brain. This metal forms stable complexes and chelates with organic molecules, nucleotides, DNA, RNA, and various proteins (enzymes). Several copper-containing enzymes include cerulo- 51 plasmin, cytochrome oxidase, monoamine oxidase, tyrosinase, super- oxide dismutase, ascorbic acid oxidase, uricase, dopamine-B-hydroxy- lase, galactose oxidase, and g-amino-lenulinic acid dehydrase. Copper's ubiquitous presence in nature allows generous intakes, usually in excess of the minimum daily allowance of 2 mg. Once absorbed through the lumen of the stomach and small intestine with the aid of a copper-binding protein (intestinal metallothionein), copper binds with albumin or amino acids for transport to the liver. Ingested copper disappears rapidly from the plasma and concentrates in the liver where a secondary increase in plasma copper occurs in the form of ceruloplasmin. As with zinc, copper levels in mammalian systems constitute wide variations due to nutritional, hormonal or pathological states. Table 7 summarizes the major metabolic conditions in which copper levels are known to play a role. . Interactions between copper and zinc have long been recognized in animals and man, as an excess of one is often associated with diminution of the other in bodily fluids and the liver. Competition for binding sites on metallothionein or metallothionein-like protein provides the best explanation. Klevay (1974) supported an hypothesis that a high zinc to copper ratio increased the risk of ischemic heart disease and associated hypercholesteremia. Although interest has been stimulated, specific human-oriented research has been lacking. Human allowances for copper must take into account variability in absorption (40-60%), reabsorption of biliary copper and the chemical state of copper in food systems. The chelating effects of fiber, dietary phytates, sulfates and vitamin C have yet to be elucidated sufficiently. Although little is known on copper bio-availability, 52 states of hypocupremia in man are relatively rare, except for the high urinary losses of ceruloplasmin associated with the nephrotic syndrome in children. Paradoxically, states of hypercupremia, due mainly to high ceruloplasmin levels, are commonly observed in pregnancy, oral intake of contraceptives, and innumerable disease states and disorders. Manifestations of copper deficiency, when they do occur, include anemia, intermittent neutropenia, severe osteoporosis and pathological fractures. Premature human infants are remarkably sensitive to poor copper intake and usually display the deficiency symptoms, described above, within two or three weeks after birth. Copper, as compared to the other trace elements identified in Table 8, has a relatively low L050. Considering that the total body stores of copper in man approach 1 gram, an L050 of 0.05 gm/kg 8W (mouse) implies a narrow margin of safety in administration of copper supple- ments. Although small amounts of the ion (15-20 mg) may cause vomiting and diarrhea, humans and pigs are known to adapt to higher levels with rapid recovery from accidental overdoses. 53 Table 5. Clinical conditions leading to Zinc deficiency Medical/Biological Condition Etiology 1. Alcoholism: induced Hyperzincuria 2. Cirrhosis of liver: low serum zinc & hyperzincuria 3. Renal disease: low plasma concentrations 4. Dialysis: low plasma concentrations 5. Burns and skin disorders: loss of zinc in exudates, ulcers, causing low plasma zinc 6. Diabetes: Hyperzincuria 7. Pregnancy and oral contraceptives 8. Acrodermatitis enteropathica: cachexia major cause of death Unknown. Effect on renal tubular epithelium postulated Unknown. Zinc salts may have protective effect on rat liver Proteinuria and failure of tubular reabsorption Unknown Skin contains 20% body stores of zinc, depletion through losses Unknown Zinc uptake by fetus; redistri- bution of zinc from plasma to RBC's Zinc deficiency due to genetic malabsorption 54 Table 6. Biochemical functions and manifestations of essential trace element deficiency. Adapted from Bland (1979) Site of Key Biochemical Signs of Element Action Function Deficiency Vanadium Hemovanadin Oxygen trans- Impaired bone & port lipid metabolism (chicks, rats) Chromium Glucose tolerance Glucose trans- Impaired glucose factor plant metabolism Manganese Pyruvate carboxy- Oxidative phos- Defective growth, lase plus many phorylation reproduction, enzymes and mucopoly- collagen synthesis, saccharide .CNS disorders synthesis Iron Hemoglobin (Hb) Hb synthesis Anemia plus some enzymes Cobalt Coenzyme 812 Biologic Pernicious anemia & Methylation methylmalonic aciduria Nickel Ribonucleic acid Nucleic acid Impaired reproduction stabilization; membrane structure (rats), deranged phospholipid meta- bolism in chicks Table 7. Metabolic conditions affecting copper homeostasis. Adapted from Evans (1973). Agent/Disease Effect on Copper Concentration Growth hormone Adrenocorticotropic hormone Addison's disease Corticosteroids Circadium variation Epinephrine Stress Estrogens and androgens Wilson's disease: (Abnormal metallothionein) Menke's Kinky Hair Syndrome Nephrotic Syndrome Depresses hepatic levels Depresses ceruloplasmin and plasma copper Elevates serum copper Reduces plasma copper Copper and ceruloplasmin's mean 24-hr. values range from a low at 1600-2400 hr. and highest at 0200-0800 hr. Elevates serum copper and ceruloplasmin Elevates plasma copper and ceruloplasmin Elevates serum copper and ceruloplasmin Excessive accumulation of copper in organs, low ceruloplasmin, low plasma copper Very low serum, hepatic, brain copper levels. (Defect in copper absorption) Low serum copper and ceruloplasmin 56 Table 8. Toxicities of selected trace minerals. Adapted from Bland (1979) Element Action Toxicity (L050) Zinc Malaise, dizziness 2.0 gm/Kg vomiting, diarrhea (Rat) Copper Hemolytic anemia 0.05 gm/Kg CNS degeneration (Mouse) Chromium Cr VI more toxic than 0.18 gm/Kg Cr III. Hyperemia, (Rat) emphysema, bronchitis Manganese Effects CNS, cramps, 0.31 gm/Kg tremors, hallucinations, (Mouse) renal degeneration Iron Hemosiderosis 0.9 gm/Kg Hemochromatosis (Mouse) Cobalt Polycythemia, abnormal 0.5 gm/Kg erythrocyte growth (Rat) Nickle Dermatitis, respiratory 0.8 gm/Kg disorders, reduces activities (Dog) of Kreb's cycle enzymes REFERENCES Bland, J. Trace Elements in Human Health and Disease: 11. The Physiologic Function of the Essential Trace Minerals. Osteopathic Medicige,7, 33-43, 1979. Evans, G.W. Copper Homeostasis in the Mammalian System. Physiological Reviews, 53, 535-570, 1973. Mason, R.E. A Conspectus of Research on Copper Metabolism and Requirements of Man. J. Nutr.,109, 1079-2066, 1979. Prasad, A.5. Clinical, Biochemical, and Pharmacological Role of Zinc. Ann. Rev. Pharmacol. Toxicol.,20, 393- 426, 1979a. Prasad, A.5. Trace Elements: Biochemical and Clinical Effects of Zinc and Copper. J. Am. Hematology, 6, 77-87, 1979b. 57 C.2. Hair, Plasma and Saliva as Indicators of Marginal Zinc/Copper Status Symptoms of marginal zinc deficiency have been identified in several segments of the population, both in the 0.5. and abroad. Typically, these symptoms include growth failure, dermatitis, inter- mittent hypogeusia, delayed wound healing and hypogonadism (Sandstead, 1973). Unfortunately, identifying marginally deficient subjects through biochemical and/or clinical testing remains difficult. Four of the most commonly used parameters--saliva, hair, serum, and erythrocyte zinc--seem to have their own proponents armed with viable arguments concerning the efficacy of use. A recent focus of attention on the use of a biological fluid on the determination of marginal zinc status is saliva. The single greatest proponent of saliva usage is Henkin (1978), who proposes a mechanism for hypogeusia based on low zinc levels in the saliva with a concomitant drop in a zinc protein (gustin) alleged to have taste bud promoting properties. Henkin and his associates were first to notice the presence of zinc in saliva following labeled 65Zn intravenous administration. Henkin cites many of his own studies demonstrating a reduced level of salivary zinc associated with taste loss in a variety of medical and/or metabolic disturbances (Henkin, 1975; McConnell et_el,, 1975; Henkin e£_el,, (1976). Henkin suggests that metals, particularly zinc, appear to be involved at several levels of organization in the taste system, more specifically, the taste bud receptor. Zinc ion may even be involved in the structure of mouse 75 58 59 Nerve Growth Factor, a compound with the ability to facilitate taste bud function (Pattison and Dunn, 1975). Unfortunately, Henkin's enthusiasm over zinc status in saliva finds little agreement with other researchers. Everett and Apgar (1979) conducted three experiments with rats to determine whether salivary zinc concentration is a more sensitive indicator of zinc status than plasma zinc. Salivary zinc concentrations in the deficient rats did not differ from those of the zinc supplemented ad libitum-fed controls in any of the experiments. While decreases in serum, bone and fetal zinc indicated a severe zinc deficiency, salivary zinc failed to reflect these changes. The authors conclude that plasma zinc is still the best available indicator of zinc status. Greger and Sickles (1979) collected mixed saliva from eight adolescent females participating in a metabolic study and 59 adolescent females participating in a nutrition survey. The zinc content of whole mixed saliva and the supernatant of the saliva was determined with a graphite-furnace AAS. Significantly (p<0.05) lower levels of zinc were found in the supernatant samples (30.5 ng/ml) than in the whole saliva samples (173 ng/ml) when the same subjects were fed 11.5 mg zinc daily rather than 14.7 mg zinc during the metabolic study. Although the levels of zinc and protein in whole mixed saliva and supernatant were correlated, the levels of zinc in both types of saliva samples were not correlated to serum or hair zinc levels of girls in the survey. The authors conclude that the use of supernatant saliva to assess zinc status may have potential. Whole mixed saliva, however, did not adequately reflect dietary status. Although Greger and Sickles (1979) expressed concern over their lack of control over flow rates, Tuompo 60 _t_el, (1977) fbund no correlation between salivary rate of flow and zinc levels. Geders et_gl, (1980) randomly assigned 32 young women (20-40 years of age) and 31 older women (41-78 years of age) into three groups, supplementing them with O, 15, or 50 mg zinc. After two months of zinc therapy, an increase in plasma zinc was proportional to the quantity of supplemental zinc. The young women exhibited a response that was approximately double that of the elderly women (67% vs. 37.5% increase over the initial plasma values for the 15 and 50 mg group, reSpectively). Maximal plasma zinc levels for the older women were attained at the 7.9 week timespan versus 5.5 for the younger women. The authors concluded that increases in the zinc content of saliva and hair in response to the two month supplementation period were not significantly different between the two age groups. In a similar study, Buchanan e£_gl, (1980) studied the response of zinc status parameters to zinc supplementation in older women (41-78 years). Only after the eighth week of zinc supplementation (15 and 50 mg zinc) were there significant differences in the zinc content of saliva and plasma. Significant increases in salivary zinc levels were noted one month following the 50 mg dosage. Of interest was the lack of change in hair zinc levels following zinc supplementation. An earlier report by Baer and King (1979) monitored zinc levels in saliva, sweat, whole blood erythrocytes and feces in the experimental depletion of zinc in young men. None of these parameters demonstrated any consistent changes with depletion over nine weeks. The results suggested that the response to low dietary zinc varies greatly among 61 individuals. Snowden and Freeland (1979) found a circadian rhythm of zinc in saliva. Highest values of zinc were noticed in late morning, decreasing to the lowest levels in early morning. No correlations were found between the dietary intake and zinc content of saliva. Johnson et_gl, (1979), in contrast to the paper by Snowden and Freeland (1979), found a decline in zinc levels of saliva after consump- tion of a low zinc diet (3 mg/day) for 21 days. When the subjects were given 50 mg zinc as ZN S0“, zinc levels in saliva increased significantly, demonstrating to the authors that saliva may be a sensitive parameter for the assessment of zinc status in man. However, their levels of zinc in saliva (126 ppm) were much higher than those found by other researchers, such as Greger and Sickles (1979), who found 173 ppb. Another concern of zinc measurements is the problem of what frac- tion of the whole saliva to analyze. Mathur e3_el, (1977) measured the relation between the zinc content of saliva and blood in healthy human adults. First, the authors noted that the mean Zn content of resting mixed saliva was one third of that in plasma and about one fifteenth that of whole blood. Secondly, the concentration of Zn in stimulated parotid saliva is only one tenth that in resting mixed saliva. In addition, nine-tenths of salivary Zn occurs in the portion of saliva containing the bulk of the mucopolysaccharide material. Finally, it is suggested that a true picture of Zn status in the body requires a knowledge of the concentrations of both Zn and Cu in the plasma. Spencer and Samachson (1968) concur with the finding of Mathur et 91, (1977). These authors found that the concentration of Zn in parotid saliva and in mixed saliva varied greatly in the different 62 patients. Not only was there great variability in the Zn levels in replicate samples of parotid and mixed saliva throughout the day, but the concentration of Zn in mixed saliva was consistently higher than in the parotid sample. The authors note that magnesium and calcium were found to have relatively constant concentrations in different samples of saliva obtained from the same person. Hair and Zinc/Copper Nutritional Statue The question of using hair biopsies to assess marginal zinc/ copper status is as controversial as the use of saliva. 0f greater importance, however, is the question of whether hair mineral analysis reflects dietary intake of the same minerals. In a most recent review of zinc status and elderly Black Ameri- cans,(Wagner e5_gl, 1980) summarized the literature concerning the ' efficacy of hair samplings. In the elderly, at least, the available information suggests that hair may be a useful index of zinc nutriture (Greger and Sciscoe, 1977; Greger, 1977; Greger and Geissler, 1978). In other individuals, however, the use of hair for the evaluation of human zinc nutriture remains somewhat controversial (Solomon, 1979; Hilderbrand and White, 1974; McKenzie, 1978). These authors suggest that contamination from exogenous sources, beauty treatments, and variable hair growth rates may complicate the interpretation of hair zinc data. Other evidence from animal studies has demonstrated that hair zinc concentrations reflect chronic or long term zinc nutriture and that it correlates with the zinc content of bone and diet (Reinhold e£_gl,, 1968; Lewis et_gl:,1957). Depressed hair zinc levels in children and adults have also been associated with human zinc 63 deficiency (Hambridge et.el,, 1972, Strain et_gl,,1966). Flynn et_el, (1971) have emphasized that hair can be of great useful- ness in nutritional assessment, if attention is given to proper sampling techniques and appropriate statistical analyses. Researchers generally agree that homogeneity in the distance of the sample from the scalp is an important consideration and that only that portion of the hair closest to the scalp should be sampled for zinc analysis. This newly grown hair is presumably more reflective of current metabolic status and is less subject to contamination from exogenous sources. The measurement of serum or plasma zinc is also frought with difficulties in interpretation and biological relevance. Although a depressed concentration of circulating zinc in plasma or serum has been associated with human zinc deficiency (Hambidge e§_gl,, 1976; Halsted et’gl,, 1974; Baer and King, 1978; Prasad e£_el,, 1978), circulating zinc levels are not always a reliable index of zinc nutriture (Mertz, 1975; Solomons, 1979; Baer, etngl., 1978). Numerous factors which are known to affect zinc status include: hemolysis, hypoproteinemia, hyperzincemia, infection, weight loss, and disease states. In addition, it should be pointed out that normal plasma zinc levels do not necessarily imply adequate body stores of this nutrient, because in some individuals plasma zinc values remain in the normal range even when the skin lesions of zinc deficiency are apparent (Baer et_el,, 1978). The variety of studies on zinc nutriture and the relevance of hair biopsies in its assessment are listed in Table 9. .Hs ocn\m= an mo=~e> ~H< 6 onmumofimaom .amm he "monEom can "moan: .Egm mm ”meme: .noamaom on .oxmucn xnauonc no m~o>o~ "so "no ecu ones Ha .mnaox Awnmnv ennom nonuno can: Ham: open mennmonuaom .anm mam "monoaom em on on muoonnamv none: -onnoo no: one mnmnosna nnm: mm" "mono: .smm bun "mono: .mwsnv ecu .xom use .omm can: vommonooo :N nnm: ":N ucN .owm one mucoEono oomne wcnaooo .amnnonmuoa oenu mo oumum can mmommm on «on was .umn ogu annmnv now xocononmoo :N on one so none: mm Homom: mn cw nne: .uono .mumn one can: omcmgo no: one an «amend .amm anm ucN an maneum ocnn nnm: wcnaooa .wnm onumocmmno a on fiasco nnw: an.mu:oEoHo ooanu nozuozz amuoonasm cuss: mo meow“ nanm~v A. ammunsoo mn on use .oman .ama mom ”:N neonMnmmmHo xcmav nne: amm.mm one meannmnnm> nanon>nccm .amm an "no amass an mucoaono oomnn mannnm .msueam ucosono -oomnu co canumenowcn now ofinmnnoncs one onomonogu use enuncnoou can“ .nnm: Annmnv -mnmmonm on» o» o>nunm=om one nnm: an mucoEoHo page ouaonvcn mnnnmom mo ucoucou u:oso~o meonnono .smmz acomnouounnonuoiocouooe use .nmez “onenuoaoeexo: e .ammz -oomnu mo monsoooonm one ucomnouov muimconuanmmonm nnm: mo moosuoe conga vonnaum mnonuz< mcnnmez mo noommm connemm< mnzmzzou =U\=N :u\=N Amnumnmsmv mozna< 22mmm xu<= mqhuh .noamoo one oznu xnnansonunen .mouconunmov nanoane mo unoammommw ecu an unnamamm eznom nnw: mo xoaonmmm .m mqmnm sun: uouenonnoo nne: sn 5 6 no99ou .osnu nos .no99oo .muen sn neuo5 onue9o: mwmmww nom moxepsn xneuonu osu .599 man "sN mo xousn se me nne: .He no uo~o-ene9 mneuos nne: see: .599 «.9 "so sn osnn use no99ou nooeu .aesu x .99eom mo 50 N on H snsunz .599 n.o~ .mneo» mm on v .muoow nne: o~95em .ooseumnu 9Heom “msonuno9 Heumnu now nose mmv .9Heom 5onm Amnmnv can: monne> so nne: seoz .599 w.~u "so ooseumnu use no99oo nne: omunosez .muonu neonusoun can: uonne> xmueonm :u\sN mo m~o>o~ .599 on "moneeoe .amneox u on N\Hu~ nne: pee .msueum suneo: cm ”none: "so .mnnnm use mxon smug nmmmww usnuonuon9 sn os~e> onnunn c.om ”monesoe nnes pen use seas: .He no mo one so\sN mo m~o>o~ nnez .599 man "none: "sN sn mgenosna ooenn mmosmnoo .on~-en eo cacao a son: .5snom use .599 m.on~ no: .Amneox osnu nne: mo sonuenonnoo oz .599 mmnxoun " maven ma on o .muoonoSm .mouonnam season eon: mso>os :N .555 mes\man " os-o mssv muse cayenneo nmmmmw nne: nonwns ue: u: .ome can: .95 can .599 mnn\mnn Hen» muo uosmnnsosnea use .ae no omeonosn sN nne: mo m~o>og \m: m.vo no: z\9 xsuneo: sn osnu nne: sounm .msonaenusoosoo .Amneox no sn5me99onsnoo no no99oo on an .monesow any nwmmww e5me~9 «no99oo..no>«~ wsoau .599 mass .mnmosnnno anennnn .He no sonuenonnoo 0: me: onosn mo omsen "so sn no99oo nae: snoum9m mnzmzzoo =o\=N :U\=N Amnueeeemv eo=n=< zzmmm mH<= mannn Aeosesueoau .a msm soozpoo :m: omno>s9 .u.sme3 Henunsn ”so Aunanv no99oo ass .onsunnuss sN now .599 m.mm~ ”mos~e> .nen on» sn amm.mm nennoues xm9onn uoom e on nnez Hennnsn "sN onsunnuss no99ou\oan xswnsz o0\ms w.m~ “monesom .88\m: n.ms "mones now on: .amneox mm on om Hu\m: mm .moaesom em «mneox .xom no owe "oneEom no9 em on em .moHea vomv sun: omseso os ues sN use Hu\us om ”moaes mHo>oH osnu one use annmnv .ome can: momeonoou sN esmens now e5meH9 esmene use xom\om< sesuan .nooeoneen sesame nausea noes .om .o~-os an one; an an .mosse> an .eom.onmoo5\ms .ms-ss .os-o .m-o one use nne: .e5meH9 mso5e sn m.umm mome now .Nen .moH .mmneox mm on n uome canoesonnou oz .Onoen sues .mNH He oes\e= .GNs .sms .nes "memo: anaemesmv .sennouea .paonmsv e on esme~9 sn sN\umm sn sN sn e5me~9 A.599 sn Hyev Am9onn me nne: >e>o~x ..m .8 one Ba 2: $298". .esmea9 use no99oo nne: .nmfi "0mm sooznon sonnenonnoo o>nunmo9 "moses ”so .mones sesu so ono5 o>es man "mofiesom Amunsue use sonunngov munsue o~e5o9 .xom use owe Hen " e5me99. w\ws m.mmumone5o9 ano99oov Hennonea Aouanv can: monne> nne: sn no99ou "none: ”so u\ms «.mmumonez "so xm9onn e we nnez. xe>o~x .mno>o9 nnes use oxeusn osnn sooznon sonuenonnoo useo .mos~e> sn omsen Aowmng -nMnsMMm os uozosm .mHHeoon ounz .599 wan .sonunnso uouneuon nmm.mm nso: um mo mnmxaese Aneuonn .599 cm 5onm usN sn osnu nne: moon: mnzmzzoo eoxen eoxen genomeeamv eo=n=< zszmm ¢Hon nnes umm.mm. so muso9ou nnes sn sN now .599 men m use sonnnnnss oan unossnom .uonenon .one nonu use nne: sn sN .mnen sn moxensn nmwmww .nne: wo mno>on so use .599 m.mn "so snonon9 use osnu son .He no oxenen acne eoaznon :5: oz .555 case "an ens: one: an =U\=N esogenoe .599 m.mn "mnonnsou .xnne55mon9 o.Nn "so mnonu mnoonwon nnez .599 oNN ”nonnsou nwmmww .no99oo nos nan .mnowennn> 599 c.nNn usN .msensenn wo .He no sensenw sn uonon9ou sN "mnowennn> nne: sn sU\sN unossnom .nennenm Amonezv .ow owe >no> moneaoe ne .599 an .ow owe ne .ome snnz :non uomeonoou sosn NH one .599 mNn on Nn .nmneox none: .sN sn omeonoou ne 599 cu .N owe ne .599 ch cm on n .msomno9 nan nwwmww e use .ome snnz no99oo one no 599 mn on .599 monuN su\sN unnes wo .ne no sn omeonosn uozosm mone5o9 "no99oo nne: owe "none: nsonsoo neno5 ooenn Nsnnonoe mnzmzzoo :o\=N =U\:N nanomeesmv eo=n=< zsmmm mwwrm u Aoosewsse9v >cuzow asarm ficEmQB :>Hz mmxcz Ns\ns N:\ns OOZZmzum mnemws mm. Awuoov <9e 5:9 ro5oewomsm omo9 as no mu ueoem. 2m” nom.mwn.n uua. on mn.n uua. Nom.w no Nno.o sm\m Na. uN.u no mn.o uoe as" No sm\m swnr o sm\9oo 59 mos ensue“ 99.9 no No.9 Ns. uoe as" Hum sm\9oo 59. Hnwwuu sw\m wwwnm sm\99 wow :59 rows New :59 no uwsn Anoo sm\m. sm\m. :9wromn N: w: ms55ee aosnsm. oeow News: wsneoome9 rowe N: w: a no 9N9 uua. :mwe osowumwm 9m eenwoone. mwsuwe. os9 oneossnnwn. :owe nonwosm :o9 so :z: inn: one oe max. unomsm o=9 some 999 so” noeeewone. unomao N: os9 oncssws 999 noeeewmne. m99eenu N: mnonsm 9m wemm arms 99eo9. zone 9m seemsp w=9ex om N5. 68 10. 11. 12. REFERENCES Assarian, G.S. and Oberleas, 0., Effect of washing procedures on trace-element content of hair. Clin. Chem., 23/9, 1771-1772, 1977. Briggs, M.H., Briggs, M. and Wakatama, A., Trace elements in human hair. Experientia, 28, 406-407, 1971. Deeming, S.8. and Weber, C.W., Evaluation of hair analysis for determination of zinc status using rats. J. Am. Clin. Nutr., 30, 2047-2052, 1977. Deeming, 5.8. and Weber, C.W., Hair analysis of trace minerals in human subjects as influenced by age, sex, and contraceptive drugs. J. Am.Clin.Nutr., 31, 1175-1180, 1978. Epstein, 0., Boss, M.A., Lyon, 0.8. and Sherlock, 5., Hair copper in primary biliary cirrhosis. J. Am.Clin.Nutr., 33, 975-956, 1980. Erten, J., Arcasoy, A., Cavadar, A.O. and Cin, 5., Hair zinc levels in healthy and malnourished children. J. Am. Clin. Nutr., 31, 1172-1174, 1978. Geders, J., Freeland-Graves, J., King, J., and Buchanan, 0., Difference in response to zinc supplementation in young and older women. Federation Proceedings, 37, 2048, 1980. Gershoff, S.N., McGandy, R.8., Nondasuta, A., Pisolyabutra, V. and Tantiwongse, P., Nutrition studies in Thailand III. Trace minerals in human and rat hair. J. Am. Clin. Nutr., 30, 868-872, 1977. Hambidge, K.H., Increase in hair copper concentration with increasing distance from the scalp. J. Am. Clin. Nutr., 26, 1212-1215, 1973. Jacob, R.A., Levay, L.M. and Logan, G.M., Hair as a biopsy material V. Hair metal as an index of hepatic metal in rats; copper and zinc. J. Am. Clin. Nutr., 31, 477-480, 1978. Klevay, L.M., Hair as a biopsy material 11. Assessment of copper nutriture. J. Am. Clin. Nutr., 23, 1194-1202, 1970. Klevay, L.M., Hair as a biopsy material. 1. Assessment of zinc nutriture. J. Am. Clin. Nutr., 23, 284-289, 1970. 69 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 70 Krebs, N.F., Moser, D.P., Blyter, E.M., Smith, J.C. and Kozlowski, B.W., Hair zinc concentrations in small-for-age and normal sized functionally retarded children. Fed. Proc.) Ant., 2045, 1980. Lindeman, R.D., Clark, M.L. and Colmore, J.P., Influence of age and sex on plasma and red-cell zinc concentrations. J. Geront., 26, 358-363, 1971. Mathur, A., Wallenius, K. and Abdulla, M., Relation between zinc content in saliva and blood in healthy human adults. Scand. J. Clin. Lab. Invest., 37, 469-472, 1977. Murthy, L., Klevay, L.M. and Petering, H.G., Interrelationships of zinc and copper nutriture in the rat. J. Nutr., 104, 1458-1465, 1974. Petering, H.G., Yaeger, D.W. and Witherup, 5.0., Trace metal content of hair I. Zinc and capper content of human hair in relation to age and sex. Arch. Environ. Health, 23, 202-207, 1971. Reinhold, J.G., Kfoury, G.A., Ghalambor, M.A. and Bennett, J.C., Zinc and copper concentrations in hair of Iranian villagers. J. Am. Clin. Nutr., 18, 294-300, 1966. Reinhold, J.G., Kfoury, G.A. and Thomas, T.A., Zinc, copper and iron concentrations in hair and other tissues: effects of low zinc and low protein intakes in rats. J. Nutr., 92, 173-182, 1967. Reinhold, J.G., Kfoury, G.A. and Arsuanian, M., Relation of zinc and calcium concentrations in hair to zinc nutrition in rats. J. Nutr., 96, 519-524, 1968. Spencer, H. and Samachson, J., Secretion of alkaline, earth metals in saliva. In: "Secretory Machanisms of Salivary Glands." Schmeyer and Schmeyeri EDS., 101-113, 1968. Solomons, N.W., Rosenberg, I.H. and Sandstead, H.A., Zinc nutrition in celiac sprue. J. Am. Clin. Nutr., 29, 371-375, 1976. Solomons, N.W., On the assessment of zinc and copper nutriture in man. J. Am. Clin. Nutr., 32, 856-871, 1979. Strain W.H., Steadman, L.T., Lankau, C.A., Berlinger, W.P. and Pories, W.J., Analysis of zinc levels in hair for the diag- nosis of zinc deficiency in man. J. Lab. Clin. Med., 68, 244-249, 1966. 71 25. Vir, S.C. and Love, A.H.G., Zinc and copper status of the elderly. J. Am. Clin. Nutr., 32, 1472-1476, 1979. 26. Wagner, P.A., Krista, M.L., Bailey, L.B., Christako, G.J., Jernigan, J.A., Araujos, P.E., Appledorf, H., Davis, 0.6. and Dinning, J.S., Zinc status of elderly black americans from urban low-income households. J. Am. Clin. Nutr., 33, 1771-1777, 1980. C.3. Factors Influencing Zinc/Copper Absorption from Supplements Although many researchers have used oral zinc/copper supplements to alleviate clinical and/or nutritional disorders, the actual bio- chemics of absorption, as influenced by dietary foodstuffs, remains obscure. Ingestion of zinc or copper salts on an empty stomach precipitates gastric irritation, intestinal motility changes, and, in some cases, diarrhea. Although acceptance of oral doses of the metals is enhanced after a meal, absorption is not. The negative influence of breads rich in phytate, interference of copper, and interactions with other nutrients can have profound effects on serum zinc levels. In one study on the effect of foodstuffs on the absorption of zinc sulfate, Pecoud (1975) demonstrated decreased zinc absorption when subjects consumed dairy products and brown bread. When 50 mg of zinc were given with coffee, the serum zinc concentration was much lower than the equivalent dose with water. When purified phytate, phosphate and calcium were given with 50 mg of zinc, only the addition of 500 mg calcium did not induce consistently lower serum levels. Of special interest was the report of gastric discomfort after 50 mg zinc on an empty stomach in five of the six subjects. The authors concluded with the warning that the absorption of ZnSOu may be completely inhibited when the drug is given with meals containing dairy products. In another study of similar experimental design, Andersson e§_el, (1976) studied serum zinc levels in eight volunteers after oral administration of 50 mg zinc as zinc sulfate. Maximum serum levels 72 73 were obtained three hours after ingestion. Some of the healthy subjects and some of the patients reported slight gastrointestinal discomfort within 30-60 min. after swallowing the zinc sulfate. Since there was little difference between the concentrations of zinc in portal and peripheral venous blood during absorption, the authors suggested the slow passage of zinc across the intestinal wall inhibits zinc absorption. A more recent article by Oelshlegel and Brewer (1977) studied the absorption of phanmacologic doses of zinc in more detail. In one study which followed the serum zinc levels of nine volunteers over one year, considerable variability was encountered; consistently high and low levels were noticed. This same variability on elevated serum zinc values was evident upon administration of 110 mg zinc sulfate in six volunteers. As expected, plasma zinc values vary as a function of dosage, with the highest serum values obtained three hours after ingestion of 50, 25, and 12.5 mg Zn. Because zinc sulfate may not be well tolerated, the authors com- pared the efficacy of using two zinc salt alternates, zinc acetate and zinc carbonate. In a serum comparison of zinc levels, the sulfate and acetate compounds showed very similar zinc tolerance curves, while the zinc carbonate exhibited lower plasma zinc levels after three hours. The final study compared serum zinc levels after consumption of various foodstuffs. In subjects consuming 25 mg Zn, the ingestion of a single hamburger bun had adverse effects on serum levels. More importantly, many food items--celery, lemon juices, coffee, hard- boiled eggs, and milk---had marked inhibitory effects on zinc absorption. Meat, histidine, glutamine, cola, club soda, ginger ale, tea, and apples did not appear to affect zinc levels. 74 Two very recent reports add additional caveats to the proper use of oral zinc supplements. Casey et_gl, (1980) studied the variations in zinc absorption in subjects consuming a standard breakfast, infant milk formulas or Lofenalac R. The standard breakfast of white toast, butter and jam was ingested with 25 mg zinc. Plasma zinc was determined prior to and at 30 minute intervals following ingestion. Although wide variability occurred in peak serum zinc values, highest values were obtained after 60-90 minutes. Male control subjects exhibited consistently lower zinc uptake values than the females, in spite of the addition of picolinic acid or citrate designed to increase absorption. LofenalacR significantly decreased zinc absorption, which may explain the poor zinc nutritional status of many children with PKU. In the other study of the intestinal interaction of zinc and iron in man, Solomons and Jacob (1980) concluded that nonheme but not heme iron competitively inhibits the intestinal absorption of zinc in man. Fe/Zn ratios of 2:1 and 3:1, but not 1:1, resulted in progressive inhibition of zinc absorption. The authors found a Fe/Zn ratio greater than 3:1 in 21 vitamin-mineral supplements listed in the Physician's Desk Reference (1977) which suggests intrinsic inter- ference with the biological availability of zinc. . The decreased absorption of zinc after administration of a Guatemalan Rural Diet was observed by Solomons e3_gl, (1979). After administration of 25 mg zinc, serum values rose significantly the first hour. Ingestion of tortillas, bean gruel, sweet rolls and black coffee with the zinc administration significantly lowered plasma zinc. The authors suggested that dietary phytates, fiber and calcium are the prime considerations for the observed inhibition of zinc absorption. Tsai and Lei (1979) demonstrated that even with high levels of 75 cellulose in the diet (16%), the whole body homeostasis toward zinc and copper was only slightly affected. Although increasing amounts of cellulose in the diet decreased serum zinc levels by a small amount, no detrimental effect on the distribution of zinc, iron, or copper in tissues was noted. The ultimate control of metal absorption apparently resides in the small intestine, at least in the rat biosystem. Evans et_gl, (1979), using radioisotope dilution to follow zinc absorption in rats, found that zinc homeostasis in rats is maintained by zinc secretion from the intestine rather than by regulation of zinc absorption. In addition, mature rats absorbed as much or more zinc than young rats fed the same dietary level of zinc. These surprising results were also encountered by Weigand and Kirchgessner (1976) who found that rats absorb far more than the daily zinc requirement and maintain zinc balance by secretion of zinc in the intestine with eventual elimination in the feces. Evans e§_gl, (1979) even goes so far as to suggest his earlier interpretation of zinc absorption was incorrect. That is, rats fed a zinc deficient diet do not absorb more zinc than rats fed a zinc adequate diet (Evans e§_el,, 1975). Mature rats simply secrete more endogenous zinc into the intestine than do young growing rats. By studying the net uptake of zinc from segments of rat duo- denum, jejunum and ileum using in vivo intestinal perfusions, the authors suggest that the capacity for zinc absorption in the rat is significantly greater in the ileum. (60.1%), at least in in vivo perfusions. Another finding of interest was the observation of rapid zinc transport across the intestinal lining upon contact with zinc. 76 It is apparent that many food items can negatively influence zinc absorption, and presumably, copper absorption when taken in the form of supplements. Therefore, the design of zinc/copper ingestion protocols or therapies must consider the types and timings of foodstuffs taken along with zinc. Armed with the foregoing information one may conclude: 1. Zinc preparations should be given ef£e5_the meal is completed, preferably 30-60 minutes. 2. Zinc and copper, and possibly other minerals, should ggt_ be given together to minimize competitive absorption. However, since there is a delicate balance in the zn/cu ratio of the blood, a disproportional amount of zinc absorbed relative to that of copper may initiate or exacerbate nutritional and/or medical disorders. 3. As compared to zinc sulfate, zinc acetate or another conjugate may lessen gastric irritation (Oelshlegal and Brewer, 1977). 10. 11. REFERENCES Andersson, K.E., Bratt, L., Dencker, H., and Lanner, E, Some aspects of the intestinal absorption of zinc in man. Europ, J. Clin. Pharmacol., 9, 423-428, 1976. Antonson, D.L., Barak, A.J., and Vanderhoof, J.A., Determination of the site of zinc absorption in rat small intestine. J. Nutr., 109, 142-147, 1979. Casey, C.E., Walravens, P.A., and Hambidge, K.H., Variations in zinc absorption. Federation Proceedings 39, p. 1213, Abstract 2041, 1980. Evans, G.W., Johnson, E.C., and Johnson, R.E., Zinc absorption in the rat determined by radioisotope dilution. J. Nutr. 109, 1258-1264, 1979. Hahn, C.J., and Evans, G.W., Absorption of trace metals in the zinc-deficient rat. J. Am. Physiology 228, 1020-1023, 1975. Oelshlegal, Jr., F.J., and Brewer, G.J., Absorption of Pharmacologic doses of zinc. 1g; "Zinc metabolism: Current aspects in health and disease," Brewer, G.J. and Prasad, A.5., EDS., Alan R. Liss, Inc., New York, pp. 299-311, 1977. Pecoud, A., Donsel, 0., and Schelling, J.L., Effect of foodstuffs on the absorption of zinc sulfate. Clin. Pharmacol. Ther. 17,‘ 469-474, 1975. Physician's Desk Reference., Huff, 8., Editor, Medical Economics Company, New Jersey, 1977. Solomons, N.W., Jacob, R.A., Pineda, 0., and Viteri, F.E., Studies on the bioavailability of zinc in man. Effects of the Guatemalan rural diet and of the iron-fortifying agent, NaFe EDTA. J. Nutr. 109, 1519-1928, 1979. Solomons, N.W., and Jacob, R.A., Studies on the intestinal inter- action of zinc and iron in man. Federation Proceedinge, 39, 430, 1980 (Abstract 856), 1980. Tsai, R.C.Y., and Lei, K.Y., Dietary cellulose, zinc and copper: effects on tissue levels of trace minerals in the rat. J. Nutr., 109, 1117-1122, 1979. 77 0.1. Animal Studies of Altered Feeding Behavior(s) with Zinc or Copper Deficient Diets. Studies concerning the influence of zinc or copper deficient diets on taste preferences in animal model systems provide more definitive information on possible taste-mineral interactions than clinical studies. Several investigators studied the effect of a zinc deficient diet on fluid intakes, altered preferences for sodium chloride and changes in plasma and urinary zinc concentrations (Chesters and Quarterman, 1970; McConnell and Henkin, 1974;' Catalanotto and Lacy, 1977; Catalanotto, 1979). The effects of a zinc deficient diet on food intake and feeding patterns of rats were studied by Chesters and Ouarterman (1970). Rats fed the Zn deficient diet reduced voluntary food intake to 70% of the control level. In addition, the Zn deficient rats ceased to gain weight and increased their day-to-day variation of feed intakes. The addition of microgram quantities of ZnSOu to the Zn deficient rat diets produced large increases in food intake within four hours. Finally, Zn deficient rats ate more of test diet containing 6 ppm Zn as compared to the 1 ppm diet. Zn supplemented diet preference was also noted by Gordon egngl, (1980) who showed that Zn deficient rats (<2 ppm) ate significantly more of the Zn supplemented diet, whereas the zinc supplemented rats (50 ppm) and zinc supplemented ad libitum rats did not. In another early experiment, McConnell and Henkin (1974) observed changes in NaCl ingestion patterns, plasma and urinary zinc in rats fed zinc deficient or zinc supplemented diets ad libitum. NaCl preference was significantly greater in zinc deficient rats; this 78 79 alteration in preference occurred within three days of the zinc deficient diet initiation, although anorexia set in the next two days. Concentrations of plasma and urinary zinc were significantly lower in rats fed the zinc deficient diet than pair-fed rats or rats fed the zinc supplemented diet ad libitum. In a similar study, Catalanotto and Lacy (1977) demonstrated an increased intake of NaCl solutions in zinc deficient rats, compared to normal rats, presumably due to decreased taste acuity. In addition to NaCl intake, the authors studied two-bottle preference for sucrose, quinine sulfate (OS) and hydrochloric acid (HCL) solutions, since NaC1 intake may be reflective of serious pathophysiologic processes not related to taste. One group was pair-fed a zinc deficient diet supplemented with ZnSOu, 100 ppm, while the other group received the zinc deficient diet, ad libitum. Compared to the controls, the deficient rats displayed significantly increased mean preferences for the sucrose, quinine and HCL solutions. In a subsequent report, Catalanotto (1979) designed an experiment to control for postingestional determinants upon fluid consumption utilizing a two-bottle, one hour test combined with a restricted three hour eating period. Zinc deficient rats demonstrated significantly greater preference for all the tastant-containing solutions (0.15 m NaC1; 0.0025 m HCL; 0.00000128 m 05; and 0.30 m NaC1 and 0.00000128 m 05), but showed both significant increases and decreases in total volume intakes. The authors suggest that increased preferences of zinc deficient rats may be related to preingestional, i.e., taste, rather than postingestional cues. 80 Increased preferences by zinc deficient animals to tastant- containing solutions suggest a physiological aberration or alteration in the gustatory mechanism(s). Catalanotto (1978) hypothesized that if the hyperkeratosis, which was observed by other investigators (Usmanski and Meyer, 1969), extended over the fungiform and circum- vallate papilla, the pores of the taste buds might become blocked and prevent the interaction of tastant and receptor. Subsequent histo- 'logical studies (Catalanotto and Nanda, 1978) failed to confirm the hypothesis, however. While the taste buds appeared smaller, the area of the pore surface exhibited a relatively normal layer of keratotic material similar to that seen in the control animals. Although the taste buds of zinc deficient animals seemed altered, cellular differ- ences were difficult to determine. Chaudhry and Meyer (1979) noted the failure of Catalanotto's taste pore hypothesis to explain zinc deficiency and taste impairment. They suggested, however, an hypothesis of their own. Functional impairment of salivary myoepithelial cells, the authors suggest, present in zinc deficient animals, might result in an impaired flow of saliva to the taste cells. Assuming that saliva contains the postulated trophic factor of Henkin gt 313(1975), then interrupted flow rates or reduced volumes of saliva may impair normal taste cell development. Copper, in addition to zinc, may be involved in the gustatory mechanisms as demonstrated by taste preference studies and copper depletions in rats. An early study by Kare and Henkin (1969) involved the use of D-Penicillamine (D-Pen) added to the diet of rats, a drug which, at least in humans, causes noticeable hypogeusia. D-Pen, added to the 81 diet at a concentration of 2 g/kg, caused significant increases in preference for 0.15 and 0.3 M NaC1 solutions. The authors suggested the altered taste acuity may be due to biochemical alterations in copper and/or thiol metabolism. In another study, Catalanotto and Henkin (1972) elaborated on the effects of D-Pen and cysteine on NaC1 preference and copper and zinc metabolism in the rat. Rats fed D-Pen exhibited greater intakes for all NaC1 solutions, decreased serum copper and increased urinary copper and zinc excretions. Rats fed cysteine exhibited higher intakes for 0.15-.60 M NaC1 solutions, but had normal serum and urinary zinc/ capper concentrations. The results indicated that D-Pen can suppress taste acuity to an extreme degree and produce anemia and endocrine malfunction, perhaps through copper depletion. Dietary intake of thiol alone (cysteine) might also suppress taste acuity through thiol group binding to the taste bud membrane. Zawalich (1971) also found marked preference shifts in D-Pen treated rats, but concluded that since electrophysiological and behavioral thresholds for NaCl were not altered, D-Pen cannot work by changing the peripheral sensitivity of the taste bud. Zawalich suggested a mechanism of action in D-Pen treated animals that centered around the possible chelating action of the drug to copper stores in the CNS. The author conceded, however, that there has yet to be established a simple relationship between sensitive or integrated nervous activity and preference. Yamamoto and Kawamura (1971) demonstrated, using electrophysio- logical studies of chemoreception, a general inhibition in the sweet taste response upon application of 0.001 and 0.0001 M copper and zinc. 82 ions to a rat tongue. The sweet taste, elicited by 1M sucrose solu- tions, was depressed (magnitude of integrated chorda tympani response) 10 to 30% of the control response. The inhibitory effect of cupric ions was stronger than that of zinc ions in the same concentrations. Human saliva normally contains 10'5M zinc and 10'7M copper, most of it being bound, however, and not in the ionized form (Dreizen _e_t_:_ 31,, (1970). The authors concluded by noting a difference in the inhibitory mechanisms of the ions; cupric ion competes with the sugar molecule for the same receptor site, while the zinc ion acts non- competitively. Elaborating on their earlier observations, Yamamoto et 91,,(1978) studied taste preference and chorda tympani nerve responses of rats under copper toxicosis. Injection of 450 ug CuCIz/IOO 9 BW for two weeks increased preference for tartaric acid and quinine solutions, but not for NaCl or sucrose solutions. Discontinuation of copper administration restored the altered preferences to the preference level of the control period. Summated taste nerve response to varying concentrations of the four basic taste stimuli in the copper-injected rats was similar to those in the control rats. Several possible mechanisms may exist to explain the observed changes in preference behavior. First, the lack of chorda tympani changes to copper injec- tion does not exclude peripheral changes that might be present in other gustatory constituents. Second, single taste fibers in copper- injected rats may have changed without any manifestation of altered whole nerve activity. Third, copper may exert its effect indirectly by influencing enzyme activities or other biological catalysts. 83 Catalanotto (1978) best summarized the influence of metal ions (notably zinc) on taste preference: ”The animal studies with various drugs seem to indicate that thiols as well as agents that deplete trace metals such as zinc cause an increased intake of certain solutions distin- guished primarily by their taste. We inter- pret this to mean a change in taste funtion. Animal studies with a dietary induced zinc deficiency . . . cause an increased intake of tastant-containing solutions and we again interpret this as modified taste function. . . Depletion of zinc can lead to decreased taste acuity but decreased taste acuity is not necessarily assoCiated with depletion of zinc." 10. 11. REFERENCES Catalanotto, F.A., and Henkin, R.I., Effects of thiols on sodium chloride preference and capper and zinc metabolism in the rat. Am. J. of Physio., 222, 1594-1598, 1972. Catalanotto, F.S., and Lacy, P., Effects of a zinc deficient diet upon fluid intake in the rate. J. Nutr., 107, 436-442, 1977. Catalanotto, F.A., and Nanda, R., The effects of administration of a zinc deficient diet on taste acuity and tongue epithe- lium. J. Oral Pathol., 6, 211-217-, 1978. Catalanotto, F.A., The trace metal zinc and taste. J. Am. Clin. Nutr., 31, 1098-1103, 1978. Catalanotto, F.A., Alterations of short-term tastant-containing fluid intake in zinc deficient adult rats. J. Nutr., 109, 1079-1085, 1979. Chaudhry, 1.M., and Meyer, J., Response of submandibular gland of the rat to nutritional zinc deficiency. J. Nutr., 109, 316-320, 1979. Chesters, J.K., and Quarterman, J., Effects of zinc deficiency on food intake and feeding patterns of rats. J. Nutr., 24, 1061-1069, 1970. Henkin, R.K., Lippoldt, R.E., Bilstad, J., and Edelhoch, H., A zinc protein isolated from human parotid saliva. Proc. Nat. Acad. Sci. U.S.A., 72, 448-494, 1975. Gordon, E.F., Denny, M.R., and Bond, J.T., Behavioral and physiological effects of experimental zinc deficiency in young and aging rats. Federation Proceedings, 39, #1, p. 431, 1980. Kare, M.R., and Henkin, R.I., The effects of D-Penicillamine on taste preferences and volume intake of sodium chloride by the rat (33925). Proc. Soc. Exptl. Biol. Med., 131, pp. 559-565, 1969. McConnell, 5.0., and Henkin, R.I., Altered preference for sodium chloride, anorexia, and changes in plasma and urinary zinc in rats fed on zinc-deficient diet. J. Nutr., 104, 1108-1114, 1974. 84 12. 13. 14. 15. 85 Osmanski, C.P., and Meyer, J., Ultrastructural changes in buccal and palatal mucosa of zinc deficient rats. J. Invest. Dermatol., 53, 14-23, 1969. Tamamoto, T., and Kawamura, Y., Inhibiting effect of cupric and zinc ions on sweet taste response in the rat. J. Osaka Univ. Dent. Sch., 11, 99-104, 1971. Yamamoto, T., Kosugi, T., and Kawamura, Y., Taste preference and taste nerve responses of rats under copper toxicosis. Pharmac. Biochem. and Behav., 9, 799-807, 1978. Zawalich, W.S., Gustatory nerve discharge and preference behavior of penicillamine treated rats. Physiology and Behavior, 6, 419-423, 1971. 0.2. Zinc/Copper Supplementation in the Chemical Senses The General Population An association between trace metals and gustatory and/or olfactory function has been suggested by several authors for some time. Approx- imately 80% of all such mineral supplementation studies, with the associated sensory restorations, have been completed by Henkin or his colleagues. Although recent reviews by Catalanotto (1978) and Anonymous (1979) tend to minimize the importance of Henkin's work, the great majority of his work, that showed improvement in taste/ smell function, cannot be categorically ignored by serious researchers. -Hence, the reader is invited to come to his own conclusions by perusal of the following papers listed in Table 10. At present, the opinion of this author agrees with the observation by Catalanotto: "Depletion of zinc can lead to decreased taste acuity but decreased taste acuity is not necessarily associated with depletion of zinc." 86 87 .munosmonsn sonnwsmooom “99 “munosmonsh sownoonon up: n .mnmxnenu nmo9 use on9 omseso nmoneonw .mnonnsoo on osn uozosm noon .msowneonwnnsoun .omeomnu w noozm .a~u\ms new noonnoo N nonw< .nnnoone5o: xosunx omenm osnn ssnom sn inomonosm .nuez use sN -uso Ann: mmnmnv ouseso os :nnz .m.h¢ .uwoe unnennen asnom .eon: Amneox monunv .mnmAnenuoao: use amm.mm uo>on95n mnmxnenuoso: .osnsnsou be .osnsnneonu mnsonne9 an znnsoe onmeh ounsm .ensmo9xs use enwzomo9x: .enxonose uonnnnsxo mnsonne9 .oosenmnsm .5snsonom w onmen oso nmeon .muosnos .s5 .eo .ua .mnoonnsm .mnmosnnno ne sn nnosm use onmen m.snxso: an .50 .sN ssnom nonnsoo 5 sn xnnsoe nwmmww now m.nzw n9 nozon nno5m use monmen .mnmosnnno mnsonne9 anonoewno .He no ue: nsonne9 seem onmea wo 59 use 99 so>on9 xm9owm ones a use Anonenmso gonna .sennansnaco .nss can\ma ens -mne .omnases on 999 nso: sN ssnom .mmosnuunu .eomses .msonnenne> .9599 can on many .osn ".99sm wo mnoowwo ounz uozosm sN nne: ..osn oxensn ounm .mxooz u now sN asnom "onoz onnoneo uoow .uo>on95n goo: x n .aomsN .sN nnez .n9nov .mnsonne9 onnno99< .x9enosn sN . N5 sue no>o .oxensn onnoneo .mneox ominN mwmxnenu onsonso amm.mm nonwe uo>on95n nonmen -mmono usnnnuonnsoo .sN asnom uoue 990 cN sn enmsouo99= nonhusnxn< msonmsnosou mnsoaonsmeoz mnsosonsmeo: mnoonssw wusnm wo o9>n nosns< anomsom neunuononm .nno5m use onmeh wo sonnesne>m xnomsow sn sonnonenoz nenoz ooenn wo xo>nsm nenosou < .sn onoen H 88 .sN us .sonn9oono9 mmosnoozm .nonu .nnes can .cm .mn .c nbmmww o>on95n xes use e5men9 .monmen unmen now e>nnem .uoonn znnsoe onmen .ne no sN .osw .99sm sN wsnnmon onmsennn onosz .e5men9 .so5o3 mssor so .99sm .oan se5uonn9 .mnonnsoo me soaoz on use so5 .momseso .enxonose .uonos Nu .msnsa onmen on uonenon ue: mnsonne9 osnns sn sN oonmou unn on amnnosenos osnn m .enmsome ues uomeonosw on usN :nnz .amnnonenos . uononn< .mnsonne9 mnsonne9 n .Aeon: .osnn: Amneox on osnu uononne nmmnww snap sn :0 ne5nos .4u=.au<29 monmen use asnom on 99 sosoz :nw: enmsomo9>s .ne no nos sN ssnom uomeonooa unmen u now n: w he sn sost so + sol nn snap neanosh sosou .onmen .Amnmn .amm.mm omnsmv mnmosno9xs an: onswon use osnn so no .senon ..mm:mm ooon-onso o.onxoo: onoeooo noon ooono one moneono noeonv onxoo: neno>om .mnounomnu onmen Nsnneonn sn osnu wo mmos>nnoowwo onnso9enosn osn x: nonnon nowanv so onsnenonnn nenmno>onnsoo ogn xusnm on mnoueon uonmononsn monn>sn semeu on x~9o¢ seueu .nononmonoso w .mnmon onmen somonnns eons :nnz .nnoo os ue: .mnsno9 oNuonoom uoow: .wnnu sN e5me~9 .nsouw>o .xe5 .sonneonw -e9eo .msnusnn neon use nonnns innsoun noonnoo sonn .unoe on enmsowo9x= now nsno9 n u unnow .sonn .mneox mN .emo>nos .nesnos an nnes nonoom sonnnsmouon eamen9 on Nn Hom< enxonose sn wo nsonsoo sU\sN onmenv men .50 w sN .mnsonne9 sonnossw onmen nbmmww "nozon onoz sN uosnos m.snxso= wne>nnem use onesow uonn use .msnaenn> .ne no xnesnn: use e5me~9 we he use he nne= .e5me~9 unenn9mo: on .onenoa ooenh no9meu msowmsnosou mnso5on3meo: mnso5onsmeo: mnoonasm wusnm wo o9xh nosn=< xnomsom neonmononm noooonoooov .on onoon 89 .sN nne: no .mnonoew xnenonu .nmn "owe on uonenonnoo .omonosm .o>ev .moneaow nos Annsoe onmen use noez wo .osoo n nn .monea nn .99sm nonwe .osn sN use none: sooznon .oxensn nsonnnss H9=onu nonnsou nnes sn 3on ono: oonoso uoonow now sonnesne>o .aon owe mnona .sN m5mn wo .mnmon onusennn :so5 xeu ma .o>e9 xnnouno .xnnsoe onmen Ammanu mneu mm now .99sm noose use xnnem now use .sN nwes oneaow nn use xnnouno osn nosemnoo we uonoowwe nos be c be .xnnsoe onmeb .munooon neonuoz use ones Nn sn .99sm oan w nomono .xnn>nnnmsom nnem noo9 wno> ue: wen .mnnsue mssox wo nesn sesn .mnmon .mon95em moon we: xnnsoe onmeb onmsennn msnm: nne: .mnneoon .mmoN .zwnv .mno>on sN nne: son use mmosnnnem anenonu .nsivN mneox mu .xnnsoe onmen w nunmnv osnu wo moxensn now n: now .mnnne: uoow so .one seoa xnnouno osn sn ooomnom son ue: noonasm wnnsoe onmeh onnessonnmoso .mnoonnsm ue sonnnnnss oan w nomono .xnnsoe onmen on uonenonnoo nos onoz sN nne: .oxensn sN .xnnsoe onmen uomeonoou .omonosm.nuez .uo>nomno ue: mnoonnsm m\n wo .osoo m use moxensn nn< .5w\wsmn sonon mno>on sooznoa oowoso .m: use sN now nne: ues mnoonasm *m uoonow .mnmon uonmon mneoz .uoue .smes use oswu n9ooxo onwsennn ”an: w n99 .sN nne: .n: .mneow osn sn msnenm onescoue xnnenosom noozm use Annem ..n: .moxensn m.mh some .one nesonnnnnss use Annmnu ono: moxensn anenono now xnnsoe onmen Anenonu weuIOn .mwvn .znnv mu oxensn anenona nouonu msonmsnosou mnso5onsmeo: mnso5onsmeo: mnoonssm wusnm wo o9>h nosns< anomsom neonuononm noooonoooou .on onoon 9O . nave eon: annoo x N sosn .xoo: n .mnnne: now xnneu ooso .eon9 msnneo eon9 Amnmnu somon as on no snooono eon: moooeon ooano>nnm .mnnnes msnneo eon9 so>no .599 an e :nnz nnnu sn sonnensoa use use .onnno99e noo9 "osnu nnez uno neon N e -on99sm oan ouunnaez .Amnmwnese .monnnsoono9 sN nne: .599 on nsmnoz now mneox u umoenmsomo9x: 9 s9 \n:Mno: son on msnsos my 599 nmnusN nne; seoz .uonmon mnoonnsm use onnno99e mnoonnsm ea :omsN wo xeu\wx\m5N m sn uonne95n noo9 ue: .Amneox onnev .sonunnso neanos AAMMMHH. >2 uonne9on annsoe annsoe onmen mnoonnsm sonunnso sn osnn nnes use .He no onmen uomeonooo .uosnos 9onu n m\m .sN nnez neanos Nmn Annsoe onmeh ouunnaez .sonunnso :neenos: sn nsooo xe5 .wou sN .Amneow so on n .uomeonosn mno>on mouev.usnnmon sN nne: use xnnsoe .msonnesne>o .nsunoz wuon onmen now .sonunnso onmen uonnnesnos m.nosno5 so uomen uu\:cmsN sonunnso c sw enmsomo9xs nmmmww sN .sonunnso uxm onnno99< .snxso: m5 Nun uoms .sN nnes now use snzonu .ne no sn son xnnsoe onmen An uosnos 9onu n .osnu nnez sonunnso enn noo9 .enxonos< ouunaaez msonmsnosou mnsosonsmeoz unsoaonsmeoz mnoonnsm wusnm wo o9xh nosns< Anomsom neonmononm soooonoooov .on osoon 1|. 9 .wnnsoe onmen uonozon m9onu Howsh .oofioz oflo «.55 Soo: .xnnsoe onmen uosnsnon .bs use he .sU\sN .nsonne9 .mson nenos nonuenm :cmsu .m.n¢ w n: nonmen n09 .uosnos Anesnns esonoxa use mnonsn use uonozon ue: nsonne9 9onuum m.snxso: use 55nom. on9nnn=5 n Annsoe onmeh snxsoz .noowwo os ue: .99sm Esnoneo no noennxo .osvnssoon 9 unonwsnene9 .nesnos 9onu1m .osonson .eu ssnom we seen nozun; 9: use n: Tonnns.osos9on:n mnso5onsmeo5 mno9e> n~< .nonnnn .osnuwnx9 wo mno9e> neonsnno .929 :nnz .5mnunonxsnene9 .nsom now ne5nos use nonmen unmen we uosnnwsoo uNimn uoue -o9xsousom9 sn Awomnv 52o 85:; s. o no .. now 5 ooo no 3688 55 8688 o ooootoonooowno :35: Amy mnmonnnw .nesnos on sonnossw onmen .uonenon nneso5ns9 uosnsnon .99sm .no99oo Annsoe onmen Huey mnnnnsnne .uo99onm son: nesnos on use no99oo nesn RNHU ennssnnmxo uosnsnon non mnsonne9 mousnosoo snxso: .5snom sn aNnv e5nou .sonnennmnsnsue Anoanv womv osom sn Annsoe .snxsoz an nonmen ono>on\no99oo -ononom :nnx osnsennnonso9ns sn .mm.mm onmen uoosuon .so9un unmen wo n9 use n: .swsmen9onsnou ”mnsonne9 nu oaseso onmeb snxsoz .munosmonsn neenos uo5nowon .oscnssoon 9onuun nsosneonn osonomnsuon9 uom= .40: .eons .Awu<9 xosononwwsm .owsen ne5nos omonosm .mouzez usn neoownnoo nmwmww osn mosnn can nu< no: .nue: now .2 use <2 onesnos m nesonue sn .ne no sn xnn>nnnmsom onmen munosmonsn sonnoono: ssnom .nsmnoz n9o<9 m munosmonsn onmen snxsoz sonmsnosou mnsosonsmeo: enso5onsmeoz onoonnsm wusnm wo o9wn nosns< xnomsom neonmoHOnm Auossnnsoov .cn onoen §-.w--- Inna. q» AK». ni~I§U\.§ 92 .musn onmen nesnosne mseo5 e>nnem wo xoea .msonnenn usoosoo can; onn9mou ween :oeon nos unu wsnneom .munosmonsn noswn: ue: mnoomnsm .m.b¢ use .mmon onmen ues mnoomnsm e>nnem wo sonmmon9ou :nn: .monen .omeomnu unnone9 nmoz .enmsomo9x: use no no9e> m neon: 3onw ane>nnem snnz mnoonosm monnno99e uomeonoou .Nsnneom uoonow .oennn9e9 on .osonusxm .o5onusxm mNnmnv ue: nNm .3onw wne> m=n9 .m.nz w n: onen~e>ssonno m.sonmonm snnz m.sonwonm sn umm.mm innem uoxoen mnoonosm 9onunn o.snxso= wo monm9onm monesow 9N nno5m use onmen snxsoz .oennn9e9 onmen wo sonnennnosoe> use sown lensonmo> uonos .nun5 sonnoonm .o9oom -onon5 nsmnn sn uonos .osos9onzn .osouson onoz momseso usn -onnns .osnunnw9 .o9oomonon5 onmen onse>nomso oz wo mno9e> now sonnuono .mnonnsoo monmen v wo wsnneom m.n¢ w n: .monmen use nsunn nN Amneow no .munosmonsn xnnmsonsn uomeonoou unmea u wo oneom An uosnaexo uvnV asoeoz nnoam use onmen finnmnv use munosmonsn xnnmsonsn conic e so ono: oennn9e9 en .soa nNV use enmsouo9ns nflm.mm uone>ono ue: mnsonne9 wsnneom .5: w n: onenne>5=onnu -umnsonne9 mm onsne9onuw snxsoz .ennssnnmxo :nnz .no.no .oN as oo ooo .nz snoonna noono ooo nononu w n: uonozonv eu no nonu on nz a: so .50 use use eenoxa .osnn use wonuenm sN .nz snns uonoonnoo uouu< .uosnos sN anesnn: snns oso Anzu noxons use use on see enmsowo9>= snxsoz 9onu-» use asnom "mnoonsso N enmsowo9>= snxsoz msonmsnosou mnso5onsmeo: onsosonsmeo: mnoonnsm nusnm wo o9xn nosns< Anomsom neonuonone noooonoooov .on onoon w ...-noon - ‘0‘.- out .~I~ h 93 .oxensn uoow nonnsoo wee osnu new mnosns< .nosun: so use .mnonnsoo sezn nozon one: sN .monen 3onw xne>nnem unnone9 .ono .nononmonosu snassne .snonon9 nenon .mosxuso no>nn .unue onns .osnsnneono .eons .omoosnu .30 «EN em: on .noa cos a mNo .sN snn: uoneonn mnsonne9 Nu .e5senn nennsnm no now .osoo 5snom seoz .eo .Noo sonnoowsn Henn> .xnnsoe .m.na w 99 nonoo9 ue: .+x 5snom osom ue: onmen use .mnsonne9 sn onmen wo .mnssoo om: mnoonssm nn< oan .non9ooon munosmonsn nne uomeono .wnnsue .moonusn on: .Amneox no onmen osn wo Asmnanv -ou moxensn oan .sN onmen now uosno5 use nnnooneao: -mNo moneaow mnmwnese onon9 amm.mm wo wusnm usnns onmsnm snxsoz 9onuum .snnonuosoz mm .monea we -onon5 nomea snxsoz .mnsn uomno>on sN uouu< .sN xnesnns nosmns use sN 5snom sn .oou "uonos omne-sonnossw .nnosm .xnneno so>nm .mnmononum -mxu nnosm use onmen now onnnon m use :omsN N5 cue onsonmxm snns anmww .enxonose uomseo onmen now uosno5 .sN anesnns nzN w woo .o5onusxm .ne no oxensn osnunnmnz snxso: 9onuum use asnom mnsonne9 xnm moon oan onso< snxso: .xon95oo osnunnmn: use sN .monnnsoe uononmon sN N5 can .Aosos9 .e5mo9xs use enxonose uonsn use osonsos .enmsowo9»; uomseo -onnns .osnunnx9o .esnouononom :nns use .munosmonsn mno9e> use Aeons .oneanos mnsonne9 use nno5m use onmen use no: .omonosm n .e5nou mnoomasm neanos nmwmww uomeonosn Amvuumo .noezo monmen .sN xnesnns ononom :nns sn enmsouo9xs .ne no momemou osnunnmn: u now m.ns a n9 use ssnom mnsonne9 m use osnunnmnz snxsoz sonmsnosoo mnsosonsmeoz onso5onsmeo: unconnsm nusnm wo o9>h nosns< anomsom neonwononm auossnnsooo .on onnen - hill-III ‘ Inn-...: i A“ ‘0‘ own. \ . snooo .oooo ono5 use Ameo moon .uommso weu\u5oon osom .nmo uoonomne imnu mnnsmon osn sn so>nm .sonnosswmxu Nn wo o .mnoonnsm ono: nos uonwnoo9m .m@ oan .sonnosswmxu anoam use onmen nmnmno oN wo nn nos .mxeu nos nsn .muosnoa wo mownnuwnes nno5m\onmen snnz ensonne9 .mm mm mmwnmN osnn wne: m.snxso= uoasmm< neonmononm mnsonne9 cN sn meioan onnass Amman. .mnoos no on mN uoue soaos mm use see we .msonn .mno>on sN 5snoo .mnsonne9 sown 1e>nomno use son :nns .enmsom .uosno5 m.snxso= .sN xnesnns nosswmxu anoam xnos m.snxso= nvnmno -o9x; ue: mnsonne9 um uossmme .ssosxsa .sN 5snom use onmen men so nennonnum zooms: .nso5o>on95n onmen :nns onenonnoo sN xne>nnem sn momeonosn xnso .mnoneonusn nenosn5 me onnennonss one .onmen use mno>on osnu osmmnn onmsnm nosno no .nwes .osnns .asnom necn moneonusn snxso: osnn so nonnum aowmno .sonnenonmon onmen sn sN wo mmoso>nnoowwo osn so oosenm on: mannwweon snxsoz osn on nonnoq snxsoz 4 o..xnnsoe onmen on uonen -on nos sN osnns no .mnouo on use muoow 5snom .xnnsoe onmen nN uom: .mno>on momeonosn osnn nesnos on nno5m unnone9 .osw .an99uno w onmen wo snsnon nesnos zones mno>on now oneom can on c .sonnoonnoo osnu unnone9 seo: msn9 .snxso: wo e>nneo use .nm.vm .omso9mon onmen .usn osonssuon wwnsm monxooxson sn Home seoso .sonnosswmxu nnoam sn nsone>nsco onooen9 msnssnnm n .wsnneom onenes9mos9 sonnusswmxu \onmen so osnu wo use onewnsm sN .ousen uoonow .snxsoz osnnexn< nnoam use wusnm no>ommonu nmmmww nesnos sn so owns: wo monsuooon9 .so .sN osnns onmen snns usnnnuonnsou .ne no son me: sN 5snom see: we w n: 9onuin use ssnom mnsonne9 can .uonnaousem snxsoz msonmsnosoo mnso5onsmeo: mnsoaonsmeoz mnoonnsm xusnm wo o9zh nosns< Anomsom neonmononm neoconnoooo .on onoon .uone>ono me: no99oo 5snom .amnmosnnno onsonso now nu\m5 Na .m> Nmo .nesnos sesn nozon sN 5snom Hench .munocmonsn onmen unu om .uo>on95n mnnnn -e9o: m< .mnsonne9 on sn enmsowo9x= .sosaoo wno> enxonos< .nsunoz xuoo no oxensn 5 a o. uoow sonn9oono9 wnomsom.onnno99e so noowwo os ue: .xeu \eo .oononoono none .nnosm use onmen sn uone>ono munosmonsn nmo: .ensmomxu .enmsoumxu .enmsomo9xs ousno .neons .qoz .omonosm .aoezo wsnneom uounow use munosmonsn onmeh .onnno99e use sown u9oono9 nno5m use onmen now oneom osnn EoOn uoma .nne o noo osos9on=n osouson -onnnz .oswunnw9 --mno9e> .eon: use so: .omonosm .noez .snonon9 eonoono noonooe so u sN 5snom .<.nn> .noo .omen nes9mos9 osnn -exne snnsn -nnno "mnmon sonnossw no>n4 .sN ussoo snnsnonaonoe5 use 5 usson sn5s9n< .oxensn uoow use osnunnmns use sN anesnns use 5snom .mn:Mno3 wuom .so 5snom suns .sN asnom son so\sN osnns .omeomnu no>nn onsonso snns mnsonne9 on .mnnnn -e9o: Henn> onsoe Ann: onsonne9 NN .nem-~m oomeo monea o .ononnsoo msn9 amneox on-e~o .oonoe -ow Nn .monea en .easenn ueo: nonwe .omeomnu Ammoww no>nn sn Annsoe .He no nnonenmso snnsm nonono .5mnnoneno5 smexen9 osnn use use oononoone-s none nonooooom nonono .nno5m\onmen .snxsoz -sn m5on95>m neon9>n no9 .nz w no use 5snom mnsonne9 9N use easenn ueo: .nonuosom msonmsnosoo mnsosonsmeoz mnso5onsmeo: mnoonnsm wusnm wo o9xb nosns< Anomsom neonwononm nooeonnoooo .on onoon 96 .munosmonsn ..mnsou .Asoo mnmonsnw onmen nosmn: use sN .wnno. umonoue nonn onnmxo sn finmano nne: son ue: 9o .sN .ossnssoon 9onuum -e9eo mssn .sN usoo cu w .9o xnnsoe onmen .Mm:mm esmen9 nesnos ues 9o snxso: uonwnuoz nne: use esmen9 onnso>sn on w msnenm oan ososonom .nmneox annoy emo>nos enxonose snns 5n use .onsnenm .xnnsoe onmen .nmon nnosm wo Asmwoo omeomnu nesnos ue; .ennso oosenonon sN .so mn “son nosos anones Romano -snno9»: snns omosn .uoms oscnssuon .sN anesnns nunnso nesnos uaenwsn onsonso .mm.mm oo>o .oooonooo nn< onsooz sono-m one eanom nn noose on on monooo oonn noon: mmwssnm nzmows .uommns .osoo :oeo now Anny uozno5 Among onoom son .osn9m oennoo nnsue -noonou onmen uom: Amneow mvuano sn sossoo .wou sN .onenonnoo nos .mnonnsoo .m.msn .mos ono5 unu mosne> onmen ones on use ues mnsonne9 9o use sN esmen9 monesow on .mosne> so nesnos .monmen unmen u now .snssnne e5men9 nosn9m :nns .A9oo nwmmww nss sN 5snom ossnssoon snxso: so use amneox mnuwNo osn9m oennoo sn .ne no son ues 9o 9onuun uonwnuo: sN ssnom none-ow on sonnnnnss oan msoaonom msonmsnosoo mnsosonsmeoz onsosonsmeo: mnoomssm nusnm wo 0999 nosns< snomsom neonuononm noooonooooo .on onoon «wa.-.:~ i-bhvdw oaww nouns-5‘. 97 .onnoonose .ones oseoos mnsonne9 N .moscnssoon nsunosno>o Anonnono xnnsoe Anwmno .osnunnmnsio snns nnosm m.snxso= so .sN wnesnns n “mones onmen use osnu amm.mm use onmen sn momseso oz uomen muosnoz use ssnom -ow omono n -nnmnsun wens nonnonnoo .mnmon sonnnsmooon nouo .nu\m5 .mseos use omooso .no>e_w soxonso nNn .m> man owennoo sn moon omne use .osnsnmne om: usN esmen9 m> .m>zo msennenooo> Anmmno onos m> now sonneonw .onennno .eons .noez .nonu use nne: .so: use Am>g nmm.mm unnsoun e5on< .monmen .omonosm now .e>nnem .esmen9 .>z can msennenomo> sn mo>eno nne so noson onoz no m> m.na onusennn sn ono>on sN use m> can xnnsoe onmeh uusenoonw .sonnon9ou snns uomeono .mmosnonnns use -ou .unnone9 nos .sN moosnnnemueons .u\usem.o usN .sonn esmen9 .mmosnonnns nos use noez now eamen9 seos .unes -9oono9 onmen Anmmno .sonn9oono9 mmosnnnem Nsnneom annmsonsn sN unnone9 onnosenos uononne use .Mm MM uomeonoou sonnon9ou sN unosmonsnen9sm use esmew9 e sn so5 m sonno~9ou oan nsunn: nomno oonooosw .nooooo -one snoao anoono 5snomiux9enosn .sN use onmen snxso: sN msosowoxo wo onxoonsnxno .m.smn we sn nNmo oan use noneonusn noo9 Nm osoz asnom .onesnos cu onxoonsnxnm seamox msonmsnosoo onsosonsmeo: mnsosonsmeo: onoonnsm wusnm wo o9>n nosns< wnomsom nooneonono muossnnsouU .on owneh 98 .sonn9oono9 onmen .msmnsesoo5 sn seo5 xes nn ness onmen so mnmosno9»: Aaumnv use xnoz m.snxso: wo unos nosnen snns .onmen so .mnounomnu mnn use onmen nwm.mm nssoooe uomenn annne9 .:.n.z no xnos wo sonnenonnom wo enmes nensoono5 osb msnxsoz .uonsonno oneo nsonne9 .mson .msonnon9 Anomsom wo Romano unon9 onmen sn onewnsm monmononno neonuoz .msonson9 nounnom osnu wo oms osn use nno5m\onmen sn oneo nsonne9 eso» nnon moosesnsnmnu use eneu m.snxso= msonnso: wo Amononmn: use noononmws9 nnosm use onmen ou neg: nonn< .sno: m.onsoo= senono wo sonmmsomnu mousnosn .onmen oneumouxno .moosonowon no99oo msnusnosn sonnnnnss no99oo .sonnnnnss sn use uonoonom wo son>on uooo wo mnoo9me nne mno>oo unwonne no99oo wo monon Ases one somnous< .mnouno -mnu nnosm use onmen sn osnn usnms now mnmnxo enmen onwnnsonom .uome osn sn Annsoe onmen so oz .monuononno xnox m.nouono wo sonnsos .msonnosswmxu nnosm Amneno on9nnns5 wo one mnouno snns .wusnm usnnsuonnsou use onmen Anesnuno usnneonn 3on>om -mnu nnosm use onmen m.snxso: wo mnmxnese nswoneo sn osnu wo mmoso>nnoowwosw sonnnnnsz .osnu wo sonnon9ou :nnz uonenoomme wnnnemmooos .xnoz nos on xnnsoe onmen nesnse use sees: mson>on9 uomeonoou nss nnnsoe wo smnonnnno momnen onmen uomeonoou on ueon nosns< .onesnse use msesss seo osnn wo sennon9ou sn onmen use osnu so ono9e9 aenmno nesn mousnosoo nosns< uonoonom wo 3on>on wonnm .onmen use osnn nenos ooenn onnoseneneo enso55oo uono>oo newnonez no9e9 wo onnnn nosns< sOnnesne>m.xnomsom use lmnnooeno: so\sN wo ozon>os .nn onoen 10. 11. 12. REFERENCES Atkin-Thor, E., Goddard, B.W., O'Nion, J., Stephen, R.L. and Kolff, W.J. Hypogeusia and zinc depletion in chronic dialysis patients. J. Am. Clin. Nutr., 31, 1948-1951, 1978. Alter, M. and Seltzer, A.P. What do taste and smell disturbances tell you? Patient Care, Dec. 1, 76-88, 1974 Anderson, N.E. and Clydesdale, P.M. The many roles of copper in nutrition. Food Product Dev., 12, 39-40, 1978. Anonymous. Ineffectiveness of zinc in treating ordinary taste and smell dysfuntion. Nutrition Reviews, 37, 283, 284, 1979. Burch, R.E., Sackin, D.A., Jetton, H.M. and Sullivan, J.F. Decreased acuity of taste and smell in cirrhosis. Gastroenterology, 67, 781, 1974. Burge, J.C., Park, H.S., Whitlock, C,P, and Schemmel, R.A. Taste acuity in patients undergoing long-term hemodialysis. Kidney International, 15, 48-53, 1979. Cagan, R.H. Reply to Henkin. Nutrition Reviews, 38, 229-230, 1980. Catalanotto, F.A. The trace metal zinc and taste. J. Am. Clin. Nutr., 31, 1098-1103, 1978. Casper, R.C., Kirschner, 8., Sandstead, H.H., Jacob, R.A. and Davis, J.M. An evaluation of trace metals, vitamins, and taste function of anorexia nervosa. J. Am. Clin. Nutr., 331, 1801-1808, 1980. Cohen, 1.K., Schechter, P.J. and Henkin, R.1. Hypogeusia, anorexia and altered zinc metabolism following thermal burn. J. AMA, 223, 914-916, 1973. Freeland-Graves, J.H., Greninger, S.A., Young, R.K., Roby, M.J., Friedman, B.J. and Vann, K.L. Taste acuity in vegetarians and nonvegetarians. Fed. Proc., 40, 840, 1981. Friedman, B.J., Freeland-Graves, J.H., Han, W.S., Young, R., Schlorlemer, M. and Geders, J. Effect of zinc supplementation on taste acuity in young women. Fed. Proc., 39, 651, 1980. 99 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 100 Geliebter, A.A., Hashim, S.A., and Van Itallie, T.B. Oral L-histidine fails to reduce taste and smell acuity but induces anorexia and urinary zinc excretion. J. Am. Clin.Nutr., 34, 119-120, 1981. Greger, J.L. Dietary intake and nutritional status in regard to zinc of institutionalized aged. J. of Gerontology, 32, 549-553, 1977. Greger, J.L. and Sciscoe, 8.5. Zinc nutriture of elderly participants in an urban feeding program. J. Am. Diet.Assoc., 70, 37-41, 1977. Hambidge, K.M., Hambidge, C., Jacobs, M. and Baum, J.D. Low levels of zinc in hair, anorexia, poor growth and hypogeusia in children. Pediat. Res., 6, 868-874, 1972a. Hambidge, K.M., Hambidge, C. and Jacobs, M.A. Taste acuity, appetite, stature and hair zinc concentrations in "normal" children. Clin. Res., 20, 265, 1972b. Hambidge, J.M. and Silverman, A. Pica with rapid improvement after dietary zinc supplementation. Arch. Dis. Childhood, 481, 567-568, 1973. Henkin, R.I., Gill Jr., J.R. and Bartter, F.D. Studies on taste thresholds in normal man and in patients with adrenal cortical insufficiency: The role of adrenal cortical steriods and of serum sodium concentration. J. Clin. Invest., 42, 727-735, 1963. Henkin, R.I., Keiser, H.R., Jaffe, J.A., Sternlieb, I. and Scheinberg, I.H. Decreased taste sensitivity after D-Penicillamine reversed by copper administration. Lancet, 2, 1268-1271, 1967. Henkin, R.I. Impairment of olfaction and of the tastes of sour and bitter in Pseudohypoparathyroidism. J. Clin. Endocr., 28, 624-628, 1968. Henkin, R.I., Graziadei, P.P.G. and Bradley, D.F. The molecular basis of taste and its disorders. Ann. Intern. Med., 71, 791-821, 1969. Henkin, R.I. and Bradley, D.F., Regulation of Taste Acuity by Thiols and Metal Ions. Nat. Acad. Sci. Proc., 62, 30-37, 1969. Henkin R.1. and Bradley, D.F. Hypogeusia corrected by Ni++ and Zn++. Life Sciences, 9, 701-709, 1970. Henkin, R.I., Schechter, P.J., Hoye, R. and Mattern, C.F.T. Idiopathic hypogeusia with dysgeusia, hyposmia, and dysosmia. J. AMA, 217, 434-440, 1971. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. 101 Henkin, R.I., Keiser, H.R. and Bronzerti, D. Histidine- Dependent zinc loss, hypogeusia, anorexia, and hyposmia. J. Clin. Invest., 51, 44, 1972. Henkin, R.I., Talal, N., Larson, A.L., and Mattern C.F.T. Abnormalities of taste and smell in Sjorgrens Syndrome. Ann. of Internal Medicine, 76, 375-383, 1972. Henkin, R.I., Patten, 8.M., Re, P., and Bronzert, D.A. A Syndrome of acute zinc losses. Arch. Neurol., 32, 745-751, 1975. Henkin, R.I., Schecter, P.J., Raff, M.S., Bronzert, D.A. and Friedewald, W.T. Zinc and taste acuity: A clinical study including a laser microprobe analysis of the gustatory receptor area. In: "Clinical Applications of Zinc Metabolism," W. Pories, W. Strain, J.M. Hsu, and R.L. Woosley, Eds., Charles C. Thomas, Springfield, ILL. Ch. 19, pp. 204-228, 1975b. Henkin, R.I., Schecter, P.J., Friedewald, W.T., Demets, D.L. and Raff, M. A double-blind study of the effects of zinc sulfate on taste and smell dysfunction. Am. J. Med. Sciences, 272, 285-299, 1976. Henkin, R.I. Ineffectiveness of zinc in treating ordinary taste and smell dysfunction. Nutrition Reviews, 38, 228-229, 1980. Hussey, H.A. Taste and smell deviations: Importance of zinc. J. AMA, 24, 1669-1670, 1974. Kosman, D.J. and Henkin, R.I. Enthrocyte zinc in patients with taste and smell dysfunction. J. Am. Clin Nutr., 34, 118, 1981. Nishi, Y., Lifshitz, F., Bayne, M.A., Daum, F., Silverberg, M., and Aiges, H. Zinc status and its relation to growth retardation in children with chronic inflammatory bowel disease. J. Am. Clin. Nutr., 33, 2613-2621, 1980. Rumble, W., Aamodt, R., Johnston, G. and Henkin, R. Absorption and retention of zinc-65 in patients with taste and smell dysfunction. Clin. Res., 23, 393, 1975. Schechter, P.J. and Henkin, R.1. Abnormalities of taste and smell after head trauma. J. Neurol. Neurosurg. Peychiatry, 37, 802-810, 1974. Schechter, P. J. and Prakash, N. J. Failure of oral L- histidine to influence appetite or affect zinc metabolism in man: double-blind study. J. Am. Clin. Nutr. , 32, 1011- 1014, I979. 102 38. Smith, F.S., Henkin, R.I. and Dell, R.B. Disordered gustatory acuity in liver disease. Gastroenterology, 70, 568-571, 1976. 39. Solomons, N.W., Rosenberg, I.H. and Sandstead, H.H. Zinc nutrition in celiac sprue. J. Am. Clin. Nutr., 29, 321-325, 1976. 40. Solomons, N.W., Rieger, C.H., Jacob, R.A., Rothberg, R. and Sandstead, H.H. Zinc nutriture and taste acuity in patients with cystic fibrosis. Nutrition Research, 1, 13-24, 1981.‘ 0.3. Use of Zinc/Copper to Restore Normal Taste/Odor Thresholds in the Elderly. The literature reports two attempts to improve taste perception in the elderly. The first study involved the influence of improved oral hygiene, via a hygienist, on taste thresholds for the four basic tastes in twelve elderly institutionalized subjects (Langan and Yearick, 1976). The twelve selected subjects, members of a geriatric home and ranging in age from 52 to 86 years, underwent the oral prophylaxis for five weeks to remove gross calculus and debris from the teeth. Eleven control subjects received no prophylaxis but were visited three times a week by the dental hygienist. Detection and identification thresholds were tested by the dilution method of Dawson et_gl, (1963), in which subjects compared solutions of increasing concentration against distilled water. Subjects varied widely in their ability to detect and identify the four primary tastes, with thresholds near those for younger adults. After oral hygiene treatments, the experimental group showed signi- ficant reductions in mean detection thresholds for sucrose and sodium chloride. The eleven control subjects were not affected. The authors concluded that taste perception, particularly for sweet and salty tastes, was enhanced by improving the health of the oral cavity. Although the experimental group increased their intakes of kcals, vitamins A, 82 and 81, calcium, protein and iron, a direct relation- ship between increased taste acuity and improved nutrient intake was not shown. 103 104 The second study by Greger and Geissler (1978) studied the effect of zinc supplementation on taste acuity in 49 institutionalized elderly. One group of 25 subjects received 15 mg of zinc sulfate daily for 95 days, while the other 24 subjects received a placebo daily. Taste acuity for salty and sweet, both detection and recognition thresholds, were measured by triangle tests of water versus three concentrations of each tastant. The detection thresholds for sodium chloride (NaCl) and sucrose improved slightly but not significantly, while recogni- tion thresholds for NaCl and sucrose were unaffected. Taste acuity, according to the authors, was ggt_correlated to dietary factors, hair zinc levels, smoking habits or use of dentures. A recent study by Friedman e; 31. (1980) on zinc supplementation in young women attempted to determine if 15, 50 and 100 mg zinc dosages would influence taste acuity after 60 days. Taste acuity was measured via forced-choice triangle tests using the four basic flavor modalities of sweet, sour, salty and bitter. Although plasma zinc values increased in each supplementation group, salivary sediment and hair zinc did not significantly change within the four treatment groups. While a significant increase in taste acuity for sweetness existed in the 50 mg zinc group, the 100 mg group failed to exhibit improvement. The authors suggest that a transient rise in plasma zinc may improve the ability to taste sweetness. REFERENCES Dawson, E.H., Brudgon, J.L. and McManus, S. Sensory testing of differences in taste. 1. Methods. Food Technolggy, 17, 1125-1131, 1963. Greger, J.L. and Geissler, A.H. Effect of zinc supplementation on taste acuity of the aged. J. Am. Clin. Nutr., 31, 633-637, 1978. Friedman, B.J., Freeland-Graves, J.H., Han, W.S., Young, R., Schorlemer, M. and Geders, J. Effect of zinc supplementation on taste acuity of young women. * Federation Proceedingg, 39, 651, 1980. Langan, M.J. and Yearick, E.S. The effects of improved oral hygiene on taste perception and nutrition of the elderly. J. of Gerontology, 31, 413-418, 1976. 105 0.4. On Determining the Proper Levels of Zinc/Capper in Taste/Olfaction Restoration Studies. In the development of the proper concentrations of both zinc and copper for supplementation purposes, several other biochemical con- siderations must be addressed: 1. Zinc inhibits copper absorption and vice-versa (Evans, 1973). 2. Klevay (1975) postulated a coronary heart disease link in diets with high zinc/copper ratios. 3. Zinc and copper sulfates may cause gastric irritation, nausea or malaise. Alternate conjugated forms of these mineral supplements may be more desirable (Oelshlegel and Brewer, 1977). 4. Estimated actual absorption of zinc and copper from dietary or supplemental sources (Mason, 1979). Most authors would agree that zinc and copper have mutually antagonistic effects on absorption. Tsai and Lei (1979), in studying the consequences of cellulose on zinc/copper metabolism, noted a small but significant increase in the tibia zinc content in rats fed copper- deficient diets as compared to those fed the copper adequate diets. This observation was explained by a physiological competition for assimilation between the two metal ions. Hahn and Evans (1975) also noted that in their studies of metal homeostasis in zinc-deficient rats, copper may antagonize zinc metabolism by interacting with the metal-binding component in the intestinal fraction. Taper e§_gl, (1980) are of the opinion that the levels of zinc and copper normally found in the diet of man have little effect on the absorption of each other. They reiterate that dietary zinc only affects the utilization of copper when the Zn/Cu ratio is excessively high (500:1). Most human diets, they contend, possess zinc 106 107 to copper ratios in the range of 10:1 to 40:1. Copper retention, they caution, may be altered in those subjects being given thera- peutic amounts of zinc for extended periods of time. The studies cited above provide supporting evidence that in zinc supplementation regimens, additional dietary copper may be necessary to prevent concomitant copper deficiencies. Certainly to be controversial is the data presented by Klevay (1975) in support of his zinc/copper ratio hypothesis and coronary heart disease. Epidemiological and metabolic data, according to the author, are consonant with his hypothesis that a metabolic imbalance (high ratio) of zinc to copper in dietary regimens is a major factor in hypercholesterolemia and increased mortality. Although the author does not state what ratio of Zn/Cu may be suspect, the article leaves the impression that ratios greater than 10:1 are involved. While the hypothesis of an elevated Zn/Cu ratio in heart disease expoused by Klevay has not met with universal acceptance, the data and arguments presented merit some attention by researchers interested in zinc supplementation and sensory restoration. Consequently, the addition of a small amount of copper seems warranted in an effort to protect the best interests of all subjects involved in the studies. Zinc sulfate, the most commonly used formula in human supplemen- tation studies, is usually associated with minor gastric irritation in doses of 50 mg to 150 mg elemental zinc (Oelshlegel and Brewer, 1977). The authors contend that alternate forms of zinc ions, i.e., the citrate, gluconate or acetate forms, may be more tolerable in patients. Hence, it behooves the experimenter to provide a source of zinc and copper that minimizes the problems of gastric irritation 108 in experiments with voluntary compliance. The final area of consideration in determining dosage levels is the percent absorption of the metal ions into the gastrointestinal tract-~notably the small intestine (ileum). Sandstead (1973) surveyed the literature on this point and found, not surprisingly, that the dietary requirement of zinc is dependent upon its availability from food and its losses from the body. Reported estimates of the bio- availability from normal diets range from 1 to 58%, although most studies report 20 to 30% as the average value. In contrast, the National Academy of Sciences has set the dietary absorption of zinc at 40 percent in their determination of the recommended dietary allowances for zinc. Despite the apparent discrepancies in the determination of zinc absorption, it remains important to recognize that oral supple- ments of zinc may not precipitate as great an increase in plasma, RBC or hair levels as the experimenter might be led to expect. REFERENCES Hahn, C.J. and Evans, G.W. Absorption of trace metals in the zinc-deficient rat. Am. J. Physiol., 228, 1020-1023, 1975. Keppel, G. "Design and Analysis: A Researcher's Handbook." Prentice-Hall, Inc., Englewood Cliffs, N.J. 658 pp., 1973. Klevay, L.M. Coronary heart disease: the zinc/copper hypothesis. J. Am. Clin. Nutr., 28, 764-774, 1975. Sandstead, H.H. Zinc nutrition in the United States. J. Am. Clin. Nutr., 26, 1251-1260, 1973. Taper, L.J., Hinners, M.L. and Ritchey, S.J. Effects of zinc intake on capper balance in adult females. J. Am. Clin. Nutr., 33, 1077-1082, 1980. Tsai, R.C.Y. and Lei, K.Y. Dietary cellulose, zinc and copper: effects on tissue levels of trace minerals in the rat. J. Nutr., 109, 1117-1122, 1979. 109 III. METHODS AND PROCEDURES A.1. Experiment I: Sensory Scaling, Salivary Flow Rates and Odor Recognition between Twenty-Five College Students and Twenty-Five Elderly Subjects. College Subjects Twenty-five Michigan State college students, both graduates and undergraduates, volunteered to be participants in the five week study, May 1, 1979 to June 9, 1979. Selection was based upon interest and availability for sensory testing. Table 20 gives the anthropometric particulars of the subjects (13 males, 12 females) who averaged 25.20 years, 174 cm in height, 65.5 kg in weight, and 16.6 in triceps fat- fold with a corresponding average of 25% body fat. Elderly,5ubjects The twenty-five elderly participants were chosen on the basis of availability and general health. Prior to selection, this author gave a short seminar to all the residents of Burcham Hills on September 5, 1979. The seminar outlined the purpose and objectives of the intended research. No attempt was made to conceal any information relating or pertaining to the intended research. Any biases that may be formed before the study by the elderly were considered in the design of the study. A flyer was prepared and distributed to ell_the residents of Burcham Hills, only after permission was granted by Mr. Redheffer, Administrator in charge of operations. Figure 1 in the Appendix shows the verbatim letter put into all the mailboxes of the Burcham 110 111 Hills residents to encourage participation. In addition, an information bulletin was distributed that outlined the particulars of the study in question (Figure 11, Appendix). Initial response was disappointing; thus, a follow-up letter was distributed (Figure 111, Appendix), encouraging the residents to witness an actual taste-testing session. With the help of Jane Greene, the activities director, and Mike Dauria, an expert in social communi- cations, additional subjects were recruited. The elderly participants also filled out a medical record form which inquired about drugs taken, vitamins used, and illnesses suffered by the participants prior to the initiation of the sensory studies (Table IV, Appendix). The intent of the study was to select a cross section of elderly subjects free from debilitating or life-threatening illnesses. According to Jane Greene, the subjects on our panel were the more enthusiastic and light-hearted members of the 240 residents at Burcham Hills. Jane mentioned to us that many of the other residents felt that their taste/smell acuity was not "good enough" for their participation or inclusion in the study. This feeling by some of the residents was later confirmed by personal observation, once Mike and I became friends with the 25 panel members. Thus, it would be important to reiterate that our taste panel did ggt_represent a true cross section of elderly subjects, but reflected a healthier, more ebullient or exuberant subsection. Table XXI gives the anthropo- metric particulars of the elderly subjects (23 females, 2 males) who averaged 83 years, 158 cm in height, and 62 kg in weight. 112 Sensory Tasting All college student taste testing was conducted in 301 Food Science Bldg. between the hours of one and four in the afternoon, three days a week, in the specially constructed taste booths which accommodated three subjects at a time. The sensory booths were constructed of particle board and painted with several coats of a latex, flat white paint. The booths were allowed to "season" for two months to insure that all the volatile components of the paint were removed. The rationale behind using 301 Food Science instead of the regular taste booths for the college students was to simulate the conditions of the elderly taste testing at Burcham Hills to be completed at a later date. The booths and all the related taste/smell apparatus were easily constructed and movable for transportation purposes. Subjects were provided with taste treats at the completion of each day of sensory testing. A maximum of three subjects at a time were allowed, with scheduled times for each participant. Subjects who were sick or missed certain testing days completed their tests the following day or at a future time more convenient for them. Elderly sensory testing was conducted from November 1, 1979 to January 12, 1980. Subjects were tested three times a week unless a holiday, such as Christmas, prevented the MWF taste testing. All sensory testing was conducted between one o'clock and four o'clock to assure the uniformity of sensory comparisons with the college subjects. The actual testing site was the "Snack Room ", a small, but very quiet room located in the basement, adjacent to the Beauty Shop. Tables and chairs were arranged such that four elderly subjects could be tested 113 simultaneously. Specially designed "Lazy Susan" booths isolated each participant during the sensory testing. Consent Forms Prior to the start of the taste studies, all panelists signed a consent fbrm, prepared specifically for this study and approved by the All University Human Use Committee, as shown in the Appendix, Figure V. PROCEDURES Experiment I was divided into five sensory parts. The studies are summarized in Table 12. A.2. Experiment 1, Parts 1 and 2. Hedonic and Sensory Scaling of Sweetness Imparted by Sucrose in Lemonade. Materials 1. Lemonade powder Powdered lemon beverage base produced by General Foods Corp., White Plains, New York. Ingredients: citric acid, monocalcium phosphate, flavoring, F D & C color, Vitamin C, Vitamin A, and a clouding agent. Lemonade provided a convenient beverage in which to vary sweetness perception by addition of sucrose. Preliminary laboratory testing established five easily distinguish- able concentration levels of sucrose to be used in hedonic and intensity testing (6, 8, 10, 14, 18% w/v). 2. Sucrose crystals High purity, (analytical reagent) grade sucrose crystals manufactured by Mallinckrodt, Inc., Paris, Kentucky, 40361. Sugar was purchased in 5 lb. containers and used when needed. r.w.: 342.30. Chemical Formula - clzuzzoll. 3. Distilled water All solutions were prepared with ion exchange distilled water using the Culligan purifying system present in the Food Science Building. 114 115 Sensory_taste apparatus Throughout the studies, 50 ml polypropylene beakers were employed which were coded with three digits using high visibility orange paint. Preparation of Lemonade Unsweetened lemonade base was prepared by dissolving 27 grams of powdered base into 3.8 liters of distilled water. Five preweighed amounts of sucrose corresponding to 6, 8, 10, 14, and 18% w/v sucrose were added. METHODS Hedonic Tastipg Judges rated degree of liking for sweetness in lemonade by circling a cross-bar on the 17-point, semi-structured hedonic scale labeled at each end with the terms "dislike extremely" and "like extremely" (Appendix, Figures VI-IX). Likewise, for sweetness inten- sity, subjects used the identical 17-point scale, but with the word anchors "extremely sweet" and “no sweetness". Between the tasting of all samples, distilled water for rinsing and a cuspidor for expectoration were provided. Serving,Conditions All solutions were presented at room temperature (21°C). The five samples were randomized and placed in aluminum serving pans prior to subject presentation. The use of randomized three-digit codes on 116 the polypropylene beakers and individualized taste booths prevented second guessing. In addition, all experiments had a trial practice run followed by three replications. If at any time the subjects had difficulty with the sensory testing procedures, the author explained the test over again, with additional trial runs if necessary. The elderly subjects had significantly more problems with the taste and smell sensory procedures. Thus, the experimenters took considerable time to explain all the testing procedures thoroughly. .mnmon zoomsom wo sownswsomou wnsw now nxon oomn 117 .msom m .no>on mswnmos nosww o use .oue -soso_ mswnmon .momuoz soson uswwwwsm .mswnmon Henna; wensou sonnou mswm: mones soww ase>wnom wo nsososamooz xooz H .m .meom m .xnwmsonsw souo wo mswweom anwmsonsw sooz n .u .meom m .mnsonouo uoow son wo sownoowwwnsouw souo xoos n .m .9>\z no.n .m.o .m.N .o.n .ov .msownosnsoosoo m .msom m .oowsu onoson sw nnem wo mswweom nnwmsonsw use unsouoz memo: N .N .9>\z son .on .so .e .oo .ooonoooooooooo o .ooos m .ouesosow sw omosoum wo msw—eom anwmsonsw use unsouo: mxooz N .n sownswnumoa seemsom sownesza .9 ausnm n.9 nsoswsosxu s9 announm use mnsousnm omownoo wo mswnmon wsomsom osn wo snesssm weowuonosossu .Nn owoon Experiment I. Part 2. Hedonic and Sensory Scaling of Saltiness Imparted by NaCl in Tomato Juice. Materials 1. Sodium chloride (NaC1) One pound containers of Mallinckrodt, Inc., analytical grade sodium chloride were purchased. Formula weight: 58.44. 2. Tomato juice All tomato juice used was the unsalted brand "Featherweight" purchased through Central Food Stores on the Michigan State campus. Nutritional information, as stated on the label, is listed in Table X of the Appendix. Preparation of Tomato Juice Fifteen to twenty 18 fl. oz. cans of unsalted tomato juice were emptied into a large carboy for proper mixing. Five concentrations of NaCl in tomato juice were used--0.0, 1.0, 2.5, 4.5, and 7.0% w/v. All solutions were prepared at room temperature (21°C) and stored in the refrigerator at 40°F (4.500) until needed. No opened, mixed tomato juice was stored for longer than three days. Methods Five samples of tomato juice with the five different levels of NaC1 were served to each taste panelist in the same manner as that described for lemonade. Three replications of hedonic testing were followed by three replications of intensity testing for each subject. 118 The identical 17-point scale was used, with only a nomenclature change of the scoresheets from sweetness to saltiness. Tomato juice hedonic and intensity scoresheets are listed in the Appendix, Figures VI and VII. Table 13. order of presentation in Experiment 1.1 Identity of foods used as olfactory stimuli and their Replication 1 Replication 2 Replication 3 H osooououna-wmo—s soy sauce pepper grape cocoa tea root beer coffee cinnamon orange onion ...: osooowmm-nwmn- root beer cocoa soy sauce pepper coffee grape tea orange cinnamon onion H 1. coffee 2. soy sauce 3. cocoa 4. tea 5. root beer 6. grape 7. pepper 8. orange 9. onion 0. cinnamon 1Only the first seven odorants were used for the intensity scaling work. Table 14. List of food odorants presented to the elderly prior to third replication of odor recognition experiment. Experiment I, Part 3.1 Soy sauce* Root beer* cherry molasses pepper* coffee* lime tuna fish grape* onion* strawberry orange* cocoa* cinnamon* garlic powder tea* 1 *Asterisk indicates the actual ten food odorants. A.3. Experiment I. Part 3. Odor Identification and Intensity of Ten Food Odorants. Protocol Rationale The purpose of part 3, Experiment I (odor identification) was to compare the olfactory responses to sniffing food odors without the use of associated sensory cues, i.e., vision, sound, etc. The criteria for selection of foods for odor identification included the purity of the volatiles produced and ready familiarity. The final choices for food aromas and their order of presentation are listed in Table 13. As with the other parts of Experiment 1, three replications were used, with care taken to keep weak odors preceding strong ones. The order of presentation of food aromas to the college students and the elderly residents was kept the same throughout the three replications. This was to assure that aroma order variance was identical in the two populations. The college students were presented with the same odorants, in different orders, for three straight days. The elderly subjects were tested by the exact same sensory protocol, except, that on their third replicate, sixteen food odorants were listed on a sheet of paper and pasted on the booth wall for the elderly to view. The theory being that if memory loss was the primary problem resulting in poor recognition scores (see results section), then the "list" might measurably improve their performance (see Table 14). Materials The olfactory set-up enumerated below was performed the morning of the day's sensory testing to ensure fresh samples for the panelists. 120 121 Each of the following materials was placed in an individual 50 ml polypropylene beaker: 1. 10. Cocoa (Nestle) powder-~loosely fill the beaker to the 30 ml mark (19 g of cocoa powder). Tea (Nestea)--place the instant tea into a beaker to the 20 ml mark and add water until the slurry levels off at the 20 ml mark. Coffee (Folger's Perk)--fill the 50 ml beaker to the 30 ml mark (14 g). Onion--mince a white onion and fill the beaker to the 30 ml mark. Cinnamon (Krogers)--fill the powder to the 20 ml level (11.5 g). 0range--cut up a fresh orange into small pieces and fill to the 30 ml level. Pepper (Krogers)--pour 20 ml of the ground pepper into the beaker (12 9). Soy Sauce (La Choy)--pour out 20 ml. . Grape (General Foods)--empty % of the grape "Kool-Aid" packet into the beaker and add water to the 20 ml mark and stir thoroughly. Root beer (McCormick)--pour enough of the liquid essence of root beer to cover the bottom of the beaker (2.5 g). 122 After the preparation of the food materials, the appropriate amounts were added to 50 ml beakers covered by aluminum foil to eliminate visual inspection. Foil lids, that had 150 pinholes punched through them, were placed on the beakers as caps. Beaker lids were securely pressed into place by pressing with thumb around the rim. All samples were allowed to come to room temperature (21°C). Four sets of the ten odorants were prepared in the morning and rotated immediately after use. A tray contained three to four odorants at a time. Each beaker was covered with a polypropylene cap to prevent aromas escaping from the hole-punched foil lids. These beakers were very effective in containing the food aromas. Subjects merely had to lift the cover, pick up the foil-lined 50 ml beaker and sniff the contents, then replace the cap. Three digit code numbers were used on the outside cap. Table X1 in the Appendix contains additional information on the food items used. The polypropylene caps were kindly supplied by the L.J. Minor Corp. of Cleveland, Ohio 44113. Preparation All food items were assembled and prepared, as outlined above, on the morning of the testing day. The onions and oranges were purchased at a local grocery store and chopped approximately two hours before the actual olfactory testing. Methods Each taste panelist received a total of ten samples, with each tray holding three to four beakers. The first seven were to be scored 123 for odor recognition and intensity, while the last three were scored for odor recognition only. The scale used was a 200 mm horizontal line in which the panelists placed a vertical line corresponding to their perception of odor intensity. The line scale ranged from no odor to high odor intensity with reference foods given for each anchor point. Milk was chosen as an appropriate reference food stimulus for a low odor food, and vinegar was chosen as a high odor food reference. Care was taken to ensure that all panelists under- stood the placement of the vertical line scale and the direction of the testing methodology. Subjects were required to sniff the reference foods to help anchor their judgments, then sniff the samples presented. First, subjects tried to identify the odor, guess if necessary, followed by the rating of the odor intensity.- As will be described in greater detail in the results section, a number of the elderly failed to perceive the aroma of the vinegar reference (about 20%). In such cases, we asked the panelists to use their odor memories of strong food stimuli, such as vinegar, for the high odor intensity reference. An example of the odor recognition and intensity scoresheets are listed in the Appendix, Figure XII. Panelists were allowed to sniff repeatedly among the samples, if necessary. Data Reduction For odor intensity, each horizontal line placed by the residents onto the scoresheet was converted to a mm number starting from the left. The 200 mm line and scores taken from it were reduced by a factor of two, thus converting the line to 100 mm. 124 The odor recognition scores were essentially a three-point scoring system. Three points were awarded for a correct choice, two points for a guess that was regarded as "close" to the correct response, i.e., cherry for grape, and one point for a response such as "perhaps it is a fruit, but I am not sure“. Zero points would be awarded for giving a response such as "tuna fish" for grape Kool-Aid. All panelists were instructed to describe the odorant if they could not name it. In the odor recognition for the elderly, the third replication used a “list" of the odorants placed before them in the taste booth. Subjects merely sniffed the samples and selected the odorant from the given list. The choice of odorants on the list is given in Table 14. In such a system, the scoring was three points for a correct response (correctly identified the odorant) and zero points for ggy- other response. The added discrimination of a full zero to three point scale was inappropriate when the list of the actual food odorants was given. It is also important to note here that the college students did not receive a list of the food odorants for their third replication. A.4. Experiment I, Part 4. Salivary Secretion to Sniffing and lasting Lemons and Lemonade. Materials Salivary Collection Egyipment 1. Johnson and Johnson No. 2, 1-1e inch cotton dental rolls. 40 ml tared plastic vials with airtight lids. Hever (W&R) stopwatch. 7 inch stainless steel forceps. 0'1th General Foods lemonade with 10% w/v sucrose (mixed from powder). 6. Fresh lemons, chopped. 7. 70% isopropyl alcohol for disinfectant purposes. Methods Whole-mouth salivary secretion was collected using three pre-weighed cotton dental rolls. One roll was bent slightly to accommodate the narrow confinement in the mouth and was placed sublingually, and the two remaining rolls were placed buccally. All subjects completed the cotton roll placement within 15 seconds, insuring a uniform salivary baseline. After the one minute and 50 second stimulus presentation, the rolls were extracted within 10 seconds using stainless steel forceps. The rolls were replaced into the previously tared plastic vials and immediately weighed to the nearest mg. When the rolls were in place, subjects were instructed to keep head and mouth movements to a minimum while carefully attending to 125 126 the stimulus. In addition, the head was kept in a horizontal plane to insure that the saliva was collected by the rolls and not pooled near the epiglottis, triggering the swallowing reflex. Each subject was tested four times. The first session was an orientation run designed to familiarize the panelists with the stimuli and collection procedures. The remaining three replications were used in the data analyses. As in all the taste sessions, food taste treats, such as cookies, were offered to interested taste panel members. A sampling of 30 Johnson and Johnson No. 2 cotton dental rolls had a mean weight of 0.348 g, with a standard deviation of 0.0296 g. Total liquid absorptivity was 2.4 g distilled water per roll. Stimuli Presentation: 1. 2 min. resting level (no stimulus present). 2. 2 min. sniffing of lemons (3 short sniffs every 15 sec., starting 15 sec. and ending 1:45). 3. 2 min. tasting of lemonade (3 drops on tongue every 30 secs., starting at 30 secs. and ending at 1:30). 4. 2 min. resting level. One minute was allowed to elapse between stimuli presentations. As mentioned earlier, subjects put the three rolls into their own mouths, while the experimenter removed the rolls 1:50 min. later. The elderly had considerable difficulty with the placement of the cotton rolls in the proper positions, as if they had "forgotten" the exact location of the oral cavity. As a consequence, it took the elderly longer to place the rolls in the mouth, particularly under the tongue, than their student counterparts. A.5. Experiment I. Part 5. Dietary Records. In addition to the sensory studies outlined above, dietary records for three days were completed by both the elderly and college students. Data were collected for three days by the college students who recorded their own intakes, while the elderly intake during meals was recorded by Mike Dauria and this author. The college students recorded their dietary intakes for three days- two weekdays_ and the following Saturday (during June 14-30, 1979), using the forms found in the Appendix, Table 13. Once the dietary records were completed, the foods were coded onto punchcards using the Michigan State University Nutrient Data Bank Code Book. The coding manual was developed under the joint efforts of Dr. Karen Morgan and Dr. Mary Zabik, members of the Dept. of Food Science and Human Nutrition. The data base itself was adapted from the HVH-CWRU nutrient data base, and is continually updated and revised as knowledge about nutrient content of food increases. If a food was not listed in the code book, then the closest food approximation was used in the analysis. If a food was a composite of several food groups and not listed in the code book, then every effort was made to find the appropriate recipe and code its food ingredients. The actual coding form used is listed in the Appendix, Table XIV. To illustrate its use, suppose a college student consumed 8 oz. of Total cereal, 1 teaspoon of non-dairy coffee whitener, 2 teaspoons of sugar, 2 cups of milk (whole), and 8 oz. of tea for breakfast. The decoder in this case would simply look up these food items in the code book, list the 6 digit code number under "item", code in the measure used (cups, 02., etc.) and fill in the 127 128 quantity, i.e., 2 cups, 2 02. etc. For granulated white sugar, the item code is 700040, the measure allowed is a teaspoon, and the quantity used is 2.00 in this example. The elderly subjects' dietary regimen was determined, for the most part, by the cyclic menu at Burcham Hills. Although the diets were somewhat restrictive, additional variance was allowable, as double entrees were a part of every meal. As the subjects sat down to their meal, we recorded all food items and intakes as closely as possible, including the post-meal weighing of unfinished foodstuffs. Portion sizes and weights were recorded prior to serving by actual measurement in the food service line. The cooperation of the head dietitian, Nancy Herbert, and food service personnel was greatly appreciated. The actual food service menu (Table XV) for the three day period is summarized in the Appendix. Subjects merely selected the desired food groups off the listing, and after a short wait, the meal was served at their own table. Because Burcham Hills had two food serving shifts, the actual recording process was considerably less hectic. Those subjects who ate meals away from the facility or who consumed late night snacks conveyed this information to us for inclusion into their dietary records. Since portion sizes are critical to calculation of nutrient content of foods, the measuring scoop sizes used to serve the meals to the elderly were carefully studied to insure accurate serving sizes (see Figure XVI, Appendix). A.6. Experiment I. Part 6. Hair Zinc/Copper Analysis. In addition to the sensory studies and dietary recalls, hair samples were obtained (0.5-1.0 g) from each participant. Hair was snipped from the nape of the neck with stainless steel scissors. Care was taken to keep the hair samples of uniform length and as close to the scalp as possible. All samples were later wet ashed and analyzed for Zinc/Copper by atomic absorption spectrometry. See "Hair Washing and Ashing" under Experiment III, in the Methods section for more details on the exact procedure. 129 8.1. Experiment 11. Second Olfactory Study with the Elderly and College Students. The decreased olfactory acuity in the elderly during Experiment 1, Parts 3 and 4, suggested a second experiment using the same elderly and college students who participated earlier. The experimental protocol of Experiment 11 focused upon the observed failure of the elderly to identify common grocery foodstuffs, such as pepper and root beer. In Part A: Subjects were required to identify ten food odorants as before; however, two of the samples were actually "blanks", containing no odorous materials. A "list" of the odorants was not provided to the panelists in this first session. In Part 8: Subjects received the ten samples as in Part A, but a "list" was posted against the taste booth wall containing the eight food items plus six additional ones. In addition, all subjects were instructed that two of the samples were blanks. A question was raised concerning the relative odor recognition abilities of the male vs. female college students participating through- out the sensory studies. In order to more closely study the potential difference, five more subjects were added to the 25 college participants, bringing the total male/female population to 15 each. Materials The olfactory set-up outlined below was prepared the morning of the taste session. All samples were purchased as fresh as possible, and disposed of after use. The following materials were placed into 50 ml polypropylene beakers: 130 131 1. Grape (General Foods)--empty 1/2 the packet into the beaker and add water to the 20 ml mark. Mix thoroughly. 2. Tuna Fish (Breast O'Chicken, Chunk Light)--lightly fill to the 20 ml mark. 3. Tomato Paste (Contadina)--fill to the 20 ml mark. 4. Lemon--dice a fresh lemon and fill to the 30 ml mark. 5. Pepper (Kroger Pure Ground)--pour 20 ml of the table pepper into the beaker. 6. Maple Syrup (Camp Pure Maple)--fill beaker to the 20 ml mark. 7. Apple Juice (Mott's Natural Style)--fill beaker to the 20 ml mark. 8. Garlic Powder (Kroger Pure)--fill beaker to the 20 ml mark. Two blank beakers (which contained only water to make the weight similar to the samples) were also prepared, in addition to the eight food samples. Table XVII in the Appendix contains the nutrient and ingredient information of the aforementioned products. The order of the odorants for recognition sessions A and B are described in Table 15. The list given to the subjects during session 8 contained the eight food odorants plus six additional food items (see Table 15). Procedure The sensory procedures for Experiment 11 were very similar to those of Experiment I. The differences in food ingredients, and list vs. without list protocol, have been outlined previously. The use of blank samples in Experiment 11 should help discriminate those subjects who “just guess" and those more positive in aroma identification. .... 132 In addition, the use of blanks allows the experimenter some insight into which "classes" of food items the subjects might mistake for actual foods. With the addition of the five college students, the age break- down is as follows: 15 males, average age 25.31:2.9; 15 females, average age 24.6::4.3. To reiterate, all college students were either undergraduate or graduate students, with additional female subjects picked from the secretarial pool. Table 15. Order of the food odorants presented to the elderly and college students in Experiment 11. First Session A Second Session 8 1. grape garlic powder 2. tuna fish ~blank- 3. tomato paste lemons 4. -blank- tuna fish 5. lemons apple juice 6. pepper tomato paste 7. maple syrup -blank- 8. -blank- maple syrup 9. apple juice pepper 0. garlic powder grape 2) Actual list of the food odorants posted inside the taste booth in Session 8, Experiment 11. tomato juice pepper apple juice peaches sardines bananas onions maple syrup grape soy sauce lemons tuna fish cranberry juice garlic powder 133 Data Reduction Scoring of the odor recognition results follows the identical guidelines of Experiment I, Part 3. Three points are awarded for a correct choice, two if the guess is reasonably close (such as cherry for strawberry), one if slightly related and zero if totally unrelated. In Part B, the panelists were presented with a list; therefore, the appropriate scoring was all or none. In other words, they either identified the food odorant correctly from the given list (three points) or failed to do so (zero points). C.1. Experiment III. The Effect of Zinc/Copper Supplementation on Odor Identification, Intensity Scaling, and Salivary Flow Rates to Food Stimuli in Eight Elderly. Subjects Eight elderly subjects, previous participants in Experiments 1 and 11, agreed to continue with the third phase of the experimental protocol--the supplementation study. Tables XVIII and XIX in the Appendix describe the handout given to all participants near the completion of Experiment 11. The actual supplementation period was from Feb. I, 1980 to March 31, 1980. This two-month supplementation period was considered an adequate length of time to allow for assimilation and absorption. As a comparison, a general survey of the zinc/copper supplementation studies in the literature finds the average length of time for oral supplementation to be approximately two months (Table 16). Egperimental Protocol Materials Randall Health Foods--zinc citrate (50 mg elemental zinc) and amino acid chelated copper (5 mg elemental copper)--were supplied to each elderly participant in the study. A sixty-day supply of each tablet was provided, along with the instructions to take the copper . tablet in the morning after breakfast and the zinc tablet after dinner. Subjects who experienced memory lapses and failed to take their tablets were not requested to "catch up" by taking two tablets the next day. 134 135 Table 16. Concentrations of Zinc/Copper and length of the supplementation period in studies using metal ions to restore sensory acuity. Supplementation Author Research Area Amounts and Length Hambidge Taste acuity in ZnSOo 2 mg/kg/day for e£_el, (1972 children 1-3 months Friedman Taste acuity in 15, 50, 100 mg zinc for et_e1, (1980) young women 60 days Atkin-Thor Hypogeusia in 220 mg ZnSOk for e; 31. (1978) dialysis patients 6 weeks. (85 mg Zn) Henkin Double-blind zinc 100 mg zinc as ZnSOt £531. (1975) Greger and Geissler (1978) Henkin 'e_t_ _a_]_. (1967) supp. study Zinc supp. and taste in the elderly Taste sensitivity and copper administration for 1-1/2 months 15 mg zinc as ZnSOt for 95 days 5-15 mg cooper as CuSOo for 4 weeks 136 Sensory Evaluation At the conclusion of the two month period, the elderly were retested on the identical sensory protocols of odor recognition, intensity and salivation, as described previously in Experiment I. Since the sensory testing of gustation in the elderly of Experiment I found little evidence of decreased taste acuity, the eight elderly subjects were not retested in this area. Furthermore, some subjects experienced fatigue along the course of the sensory retesting; the decision to eliminate the gustatory protocol was made. Hair Analysis At the beginning of the sensory testing and at the end of the supplementation period of Experiment III, hair samples were obtained (0.5-1.0 g) from participants. Hair was snipped from the nape of the neck with stainless steel scissors. Care was taken to keep the hair samples of uniform length and as close to the scalp as possible. All samples were later wet ashed and analyzed for zinc/copper by atomic absorption spectrometry, as described in a following section on hair washing and ashing. METHODS Olfactory,Testing_ At the end of the supplementation period, subjects were retested on olfactory recognition and intensity to the identical foods used in Experiment 1, Parts 3 and 4. Subjects were required to first identify the odorant and then rate the intensity. The order of the testing was: 137 I. Odor identification of the ten food odorants without a list, then an intensity rating of seven food odorants (1 rep.). 2. Odor identification of the ten food odorants gigb a list, followed by an intensity rating of seven food odorants (1 rep.). 3. Final intensity scaling of seven food odorants (1 rep.). Salivagy Testing Salivary flow rates were measured in the eight subjects, as described in Study 1. To reiterate, 2-min. salivary flow rates were measured in response to: 2 min. resting level (no stimulus present) 2 min. sniffing of lemons “NH 2 min. tasting of lemonade 4. 2 min. resting level (no stimulus present) Finally, at the end of the experiment, all subjects were reweighed and asked for any subjective feelings on whether the mineral therapy was affecting their sensory preparation. A list of their comments may be found in the Appendix, Table XXV. Hair Washipg and Ashing Hair Washing All collected hair samples were subjected to the following cleans- ing procedure prior to the wet-ashing. The procedure used was a modified version of the methodology by Klevay (1970). Hair samples were collected and finely chopped with stainless steel clippers. After chopping, the samples were placed in individual plastic vials 138 and washed according to the following: 1) acetone--3 times 2) 1% (Triton X)--3 times 3) Distilled water--3 times The washing procedure consisted of shaking the sample in the selected washing medium for 2 minutes, whereupon the solution was drained off. Individual 50 ml polypropylene beakers were used throughout the analysis to avoid metal ion contamination. After the final rinsing with dis- tilled water, the samples were allowed to dry Overnight in a calcium chloride dessicator. Wet Ashing The dried hair samples were weighed accurately to three places on an analytical balance. Care was taken in the weighing and transfer to insure against loss of small hair fibers in handling. After transfer to glass ashing vials, 3 ml of concentrated nitric acid was added to each of the tubes. Distilled water was added as a washdown along the sides of the boiling flask. Samples were then heated to boiling for approximately 30 minutes or until a small amount of clear, nonfuming liquid remained. Digested samples were quantitatively transferred to clean polypropylene beakers and brought up to 10 ml final dilution with distilled water. After the appropriate standard solutions for zinc and copper were prepared, samples were analyzed using a Hitachi 180-70 Model Atomic Absorption Spectrophotometer (AAS). The cooperation of the Carnation Company for the assistance and use of their AAS is greatly appreciated. REFERENCES Klevay, L.M. Hair as a biopsy material. Assessment of zinc nutriture. J. Am. Clin Nutr., 23, 284-289, 1970. I39 IV. RESULTS A.1. Experiment I. Sensory Scaling, Salivary Flow Rates and Odor Recognition between Twenty-Five College Students and Twenty-Five Elderly Subjects. Parts 1 and 2: Hedonic and Intensity Scaling of Sucrose in Lemonade and Tomato Juice. Subjects Twenty-five college-age subjects and 25 elderly subjects volun- teered to participate in Experiment 1. Table XX, in the Appendix, details the anthropometric data of the college participants, who averaged 25.2 years of age, 65.5 kg in weight and 25.0% body fat. The college group consisted of 12 females and 13 males, picked from a pool of graduate students and secretaries working in the Food Science Building. Table XXA, of the Appendix, describes the anthropometric particulars of the 25 elderly residents of Burcham Hills. Subjects averaged 83.5 years in age and 62.0 kg in weight. The group, whose age ranged from 74 to 95 years, consisted of 23 female and 2 male subjects. Appendix Table XXII summarizes the anthropometric data on the two subject groups for side-by-side comparison. The elderly subjects were required to fill out a brief medical survey that asked their history of drug and vitamin ingestion, smoking and current illnesses. Appendix Table XXIII summarizes the questionnaire and the findings. A majority of the subjects questioned were taking medication (76%) at the time, but only 32% were concurrently taking vitamins. Two of the subjects were diabetic, one had kidney stones, and eight were hypertensive. None of the subjects were 140 141 smokers. Table XXIV, in the Appendix, lists the drugs mentioned in the medical record of the elderly. 1. Hedonic Response to Tomato Juice Figure 1 illustrates the average hedonic and intensity response to tomato juice by the 25 elderly and college subjects. Inspection of the hedonic responses, by both groups, reveals a similar 1.0% NaCl- peaking sensory function. Because of the great variability in individual responses, the slight elevated funCtion for'the elderly would usually have little significance. However, there was a highly significant interaction by Group and Concentration (A X C) (see Table 17). This significance is suggested by visual inspection of the consistently elevated elderly response to the degree of liking of the various salt levels. Of considerable interest is the non-significant judge F-ratio (1.09) indicating good subject agreement for all concentrations. The low F-ratio for replications again highlights the ability of the judges to replicate their own sensory evaluations. 2. Intensity Reeponse to Tomato Juice A similar situation exists in the intensity ratings of the tomato juice by the two groups. Figure 1 depicts a narrower intensity range of response by the elderly (5 to 15.5 points) than the college students (1.5 to 16 points). Table (18) shows a significant F-ratio for groups (elderly vs. college) in the grand mean, individual judges, concentrations, but not in replications. The most important interaction (A X C), groups 142 X concentration, was highly significant, indicating a difference of opinion in the intensity judgements by college students and the elderly. 143 .n.9 nsoewsosxu .ouwzu onoson sw msownosnsoosou nooz mswnso> on momsosmos nnwmsonsw use owsouos soos wo somwsooeoo .n mesons 144 mi m.N .0ez omoo< xxo 0.9 on 3 me on o s n 4}) T n n 11. 1 l >.:mzw._.z_f momssooolllb 555.5 out ...o , O_ZODmI _ . «(s wx.._m.a l No.2. 0.5220... SSNOdSBH AHOSNES BQVHBAV 145 .nno>nooosoon .noo.o oso .so.ovo no noeonwnoonm .... ... .Onnenuw uonensnen osn onensoneo on uoms nonesnsosou osn wo ounsom oaks mew n noon wo oonsom mosh. onszonuumowzosm: .nun nsosnno9xm sn monsousnm owonnoo use xnnouno osn so oonsn one5on sn mmosnnnem on momso9mon unsouos osn now monnenaw use A.m.mo monesom wo sso osn .n.9.so 5ouoonw wo moonuos noosenne> wo mnmwnese uonmoz .whno onsen 146 .nnosnnoosoon .noo.o oso mo.ovs no osoonwnsenm .... .. .OMHMHIN— fiOHfiHDDdH 0:9 GHflHqfiuHmu OH 60m: HOHGEMEOG0§ 058 MO OOH—now OSHA own s neon wo oonsom mowau OHHHmszzH ..n.w nsosnno9xm sn monsousnm omonnoo use wnnouno osn no oonsn oneson on momso9mon Annmsonsn osn now monnenum use A.m.mo monescm wo ssm .A.9.oo 5ouoonw wo moonmou noosenne> wo mwmwnese uonmoz .Aeno onoen 147 3. Hedonic Reeponses to Lemonade Table (19) summarizes the results of the nested analysis of vari- ance for the hedonic responses of the elderly and college students to varying levels of scurose in lemonade (Figure 2). The hedonic scaling by the elderly followed closely to that of the college students for the first three concentration levels of sucrose (6, 8, 10% w/v), then increased slightly in the last two concentrations (14, 18% w/v). This difference was significant, but barely so (5% level). Although judges did not differ significantly from each other, the replications per judge did. 4. Intensity Responses to Lemonade Inspection of the intensity graphics to sweetness in lemonade (Figure 2) demonstrates a greater disparity in response by the two groups than to the NaCl in tomato juice. As with the NaCl graphics in tomato juice, the elderly displayed a constriction in sweetness intensity range (6.3 to 14.6%) compared to the college students (2.7 - 15.8%). Both the group and the group X concentration inter- action was significantly different, as shown in Table (20). Although the replicate samples differ from each other (F-ratio: 4.58*), the magnitude of the group by concentration interaction was so large (F-ratio: 26.23) that it overshadowed the increased sample variability. Table (21) contains the hedonic and intensity means for groups (elderly vs. college) and tastant concentrations for Experiment 1, Part 1. 148 .>~o>nnoo9mon .Hoo.o use no.o .mo.ovs no osoonwnsMnm ......... .Owumhin— woumuan—eu on; Quwmfiommu OH flow: Houmcwfiocowu 6:» HO OOH—now ”Fr H eon s neon wo oonsom on momso9mon .ouesoson sn omonosm onsouo: now mensousnm omonnoo .m> xnnounoo e>os< uonmoz .mmno onoon 149 .N.9 nsoswnoexu .ouesosow sw mwo>ow omonosm mswzse> on momsoemon znwmsonsw use unsouos omeno>e osn wo somwneesoo .N onsmw9 150 mmOmoaw omao< nxo or a 003. up or a o o I. 10.... >....mzm...z_ d _ N5 a J]. : - mx...m.oJ >..¢wo..m OI I I .6 NOW—.50 Olilll.’ 1_ 0.20nm1 . In): wv.....l mo<202mo BSNOdSHH AHOSNBS EQVHBAV 151 .nno>noooomon .noo.o oso mo.ovo no oooonnnsonm .... .. .onnena9 uonensnen osn onensoneo on uoms nonesnaosou osn wo ounsom oaks eon s amok mo ouufiom OmOHUSW OH mo<202mgai>bwmzmth nouesoson sn momso9mon annmsonsn now momonnoo .m> xnnounoo e>os< uonmoz .noN. onsen 152 Table (21). Hedonic and intensity means for groups (college vs. elderly), and concentrations (of NaCl) for the tomato juice and lemonade food systems of Experiment I. TOMATO HEDONICS LEMONADE HBDONICS A-Groups:*** A-Groups:* Elderly College Elderly College 7.11 6.05 10.78 9.81 C-Concentrations:*** C-Concentrations:*** 0.0 1.0 2.5 4.5 7.0 6 8 10 14 18 8.06 12.42 6.62 3.42 3.04 7.24 9.90 11.38 11.76 11.18 TOMATO INTENSITY LEMONADE INTENSITY A-Groups:*** A-Groups:*** Elderly College Elderly College 10.99 9.56 10.63 9.11 C-Concentrations;*** C-Concentrations:*** 0.0 1.0 2.5 4.5 7.0 6 8 10 14 18 3.38 6.71 11.48 14.0 15.58 4.76 7.22 9.46 12.94 14.98 1Underlined means within a row do not differ significantly. *, ***, Significant at P<0.05 and 0.001, respectively. 153 Of interest is the consistently higher group means for the elderly throughout the two food systems. The subjects, both elderly and college, had an easier time separating the five concentrations of lemonade by intensity, as evidenced by the greater number of mean separations, than the NaCl in tomato juice. Because the hedonic function for sweetness in lemonade is an inverted "U" shape, levels 14-18% sucrose were not separated well by the elderly panelists. Tables (22) and (23) contain the separate analysis of variance for the elderly and college students in both the hedonic and intensity scalings of tomato juice and lemonade. In the replica- tions of their individual sensory scalings, only the elderly (intensity, lemonade) had any difficulty. However, in the F-ratios for the other three food systems, the elderly matched the college subjects' low variabilities. As far as the concentration and subjects' variations are concerned, both groups displayed remarkable similarity. Figure 3 depicts the average sensory response to tomato hedonics and intensity in five classes of elderly response types. For example, subject number four, in the upper left, displays the "ideal" opposite bifurcation of the sensory functions of hedonics and intensity. In contrast, subject number 13 demonstrates a "flat" intensity curve, suggesting a decreased ability to taste NaCl in tomato juice. Five elderly subjects, in total, displayed a flat intensity curve, as seen by subject number 13. However, the same subjects did rate the hedonics of the higher concentrations in a downward fashion for all four of the "flat“ intensity curves. 154 .onnenu9 osn on 9 use 5ouoonw wo moonmou on .9.oH .nno>nnoosoon noo.o oso .so.o .mo.ovs no nooonwnoone .... ... . A uwm .s omonnoo 9nnounm omonnoo wanounm wo oonsom ouesosos oonsu one5on noez use ouesoson ..H nsosnno9xm .oonsn one5on sn sn omonosm on oeso9mon unsouos osn now >o< onene9om .ANNU onsen 155 .onnenu9 osn on 9 use 5ouoonw wo moonmou on .99H .nnonnooosoon noo.o use .so.o .mo.ovo no nooonwnounm .... ... .. one s ouesosos ounsu one5on f wo oonsom use ouesoson.sn .9 nsosnno9nm .oonsn oneson sn noez omonosm on momso9mon Annmsonsn osn now >Q< onene9ow .mmNo onoen 156 .n.. nsoswnosxu sw annouwo osn so monsouos use xnwmsonsw one5on on moszn xsomsom o>ww wo essenm zoomsom nesuw>wus9 .m ossmw9 157 .0 e2 Oman: .x. on m... on on o on. o... ...N on s on m... 3. on o n u . nH- . . . JV. . . . JV 0.. ..o/ / I Foss N x ..o a. . .525. :22 .l >....wzm...z. m0.20om1 H09.520... >..mmo..m (8 = N) SSNOdSEH AHOSNES BQVUBAV 158 This would suggest that the elderly subjects, although unable to differ- entiate the higher levels of NaCl, were still able to rate the solutions as unpleasant. Figure 4 illustrates five examples of individual sensory scaling by the college students to tomato intensity and hedonics. Note the distinctive sensory response profiles of hedonic versus intensity scaling. None of the college students had any difficulty separating out the higher levels of NaCl concentrations. Figures 5 and 6 depict the individual sensory graphs of five response types to lemonade intensity and hedonics by the elderly and college students, respectively. In the lemonade food system, ten elderly subjects exhibited “flat" intensity functions as the concentra- tion of sucrose increased. In Figure 5, subject number 18 is an example of the "flat" response. A confused response, one in which a concentration is not scaled correctly, is shown by subject number 25, in the lower right corner of Figure 5. Ten elderly subjects had some difficulty correctly scaling the sucrose concentrations in order, although this confusion is not obvious by inspection of panel average. Table (24) sunmarizes the number of elderly subjects with "flat'I or "confused" sensory scaling functions. A question might be asked concerning those subjects who fall into the flat or confused response category: 00 they repeat themselves in both taste systems? By observing Table (24), one finds that three elderly with flat functions for tomato juice were also flat in lemonade. In addition, six individuals were possessors of flat and confused sensory functions in the tomato juice and lemonade. 159 .n.. nsoswnosxm sw mnsouanm omo—noo osn as muwsouos use anmsonsw onoson on moszn omsosmon o>ww we essenm snowsom nezuw>wus9 .u onsmw9 160 .0 e2 omoo< .x. mi. ad. OJ 9.9. mi ad c.— a 6.9 n . C1 ‘\ rte ad a; 0 O 1- P“ ......s o x ..o ...N. smzn_w.zmnz_-\. oozes :. 5222 s s momjoo = N) asuoasau AUOSNES aovuanv (8 161 .N.9 nsoswsoexm sw mnuoussm xnnouwo osn 9o muwsouos use 9nwmsonsw ouososow on moean omsoemon o>ww we assess nsomsom wezuw>wus. .m onsmws 162 mmOmozm Oman: o\o e e on up an e e on 3. on _ n _ n . i-.. at. Foes o n ..o ...N. so... .325. (8 = N) HSNOdSSH AHOSNSS EQVHEAV >....m2m...z. mO.ZODwI an..mmo..m 163 .N.9 nsoswnooxm s. mnuonszm omonnoo osn no muwsouos use anwmsonsw ouososow on mosxn omsoamon o>ww wo mseenu seamsom nesuw>wus. .o 9:5: 164 mwOmozm omna< Qo 2. up 2. a o o... I. oi lo >n_ezmnz_ ...oos o x ..o .5. smz< N- s 'TrrIIUIIITlTIITIIU L [ l rL In r1 m62(¢0 {\\\\\\\\\\\ [ cummmu IL mm<¢6 N m llllllllLillllllLll IL w03Om Ill zmmn IL (uh boo: Samoa V\\\§ 33.60 I mpflsmuomzw Mmfla ..qmgflflm. do?” lllllllllllllllllllllll >._mmo._w .m> mamaaoo H20.._.< :— mug-(m >Om main-O Emma #00: 202(225 wb<400020 TIIIIIflijfT11TIITI 26.20 moza¢maaw “20.h<0.u.hzwo. 2000 (58 = N) BSNOdSSB 1038800 lNSOHBd 176 Table (27). Statistical treatment of the odor recognition of Experiment 1.3. A. Chi-Square: x2 = 33.3*** Reference: Hewlett-Packard (1974) B. Paired t-statistic: T = 16.55,*** df=9 Reference: Roessler (1975) C. Analysis of Variance: DF 83 ' MS F Total 39 34310.0 Odors 9 5160.5 573.39 6.81*** Groups 1 26832.4 26832.4 318.49*** Reps 9 632.2 70.24 0.83 Error 1684.90 , 84.25 Reference: Roessler (1975) ***, Significant at P<0.001. A.3. Experiment I, Part 3. Intensity Ratings to Seven Fbod Odorants by the Elderly and College Students. Figure 9 depicts the average intensity response to seven food odorants by the 25 elderly and 25 college students. Odorants are arranged such that the intensity differences between the two groups increase as one visually travels to the right. The food odorants--c0coa, tea and grape-~received similar odor intensities by both groups. However, two very common foodstuffs, pepper and coffee, were rated quite dissimilar. Surprisingly, pepper, known to have a strong trigeminal or pain stimulus component, was rated at almost half the strength of the college students' level by the elderly participants. This difference agrees with the previous observations by the experimenters which found that at least 20% of the elderly failed to detect the food reference for high odor intensity vinegar. In any event, the intensity rankings of the odorants by the two groups were not related, as the Spearman rank correlation coefficient is only r5=.643. The nested analysis of variance (Table 28) finds significant variation in the group means (41.8 vs. 61.4), the odorants, and judges. Replications, however, were not significantly different, indicating good agreement by all judges through the three repeat trials. The group by odorant interaction was highly significant, indicating the lack of agreement for intensity ratings of the seven odorants by the elderly and college students. Inspection of Table (29) reemphasizes the difficulty of the elderly to differentiate the seven food odorants. Using the Scheffe’mean comparison technique, a very 177 178 conservative separation technique, the elderly failed to separate many of the odorants by intensity alone. One reason for this failure to differentiate is the greater inherent error variance in their intensity judgments. The separate AOV of Table (30) reveals a significant replication factor and a significant odorant by replication factor. In addition, note the large error variance for the elderly as compared to the college students (M.S.: 242.06 vs 157.79). 179 .¢.~ acmevcmaxu eo xpcmvpm mgu .m> mmmFFoo wsu An um>_mocma .mpzs_um uoow cm>mm mo au_mcoucp coco maucm>< .m mgzmwd 180 ...—...ZFm >mOhomw hood, .3: a. l._. r 1. 30h L: I” .330 1 326 h >4mmoqm g. .. momjoo Us. 8... .. m .. a .. w .... m. S.NG 2: O 8 [.8 76M 2 m... a N 8 S . I. ..A 181 .»~o>wooooooo .Hoo.o oeo ~c.ova oo oeooumMeuflw .... ... .OMHmnum vouaaznau can oumaaogoo on com: Houmcwaocov on» we connom onh~ ova" o Home mo ransom ..v.H uaoafiuonxm mo mucovaum ouofiaou one xfluoego one an mueonoeo uoow =o>om one on noncommoa xufimcoucw on» mo >o< voumoz. .fimau oases 182 Table (29). Intensity means for groups and odorants of the seven food odorants, Experiment 1.4. 1 FOOD ODORANT INTENSITY Nested AOV: A-Groups:*** Elderly College 41.81 61.44 B-Odorants: *** Elderly Tea Cocoa Soy S. Grape Root B. Coffee Pepper 41.20 44.52 63.6 64.72 65.72 74.36 76.40 Separate AOV: Elderly Tea Soy 8. Cocoa Coffee Pepper _ Root 8. Grape 32.56 35.44 40.52 42.60 44.92 45.84 50.80 291.1282. Tea. Cocoa . 50y 5. Grape Root 8. Coffee Pepper 41.20 44.52 ‘"63.16 64.72 65.72 74.36 76.40 1Within the same row, means underscored by the same line are not significantly different. ***, Significantly different at P<0.00l. .monmacm mo 55m cues“ .NHo>Nooomooo .Hoo.o one .No.c .mo.ovo om ocoofimfiomwm .444 .44 .4 mmN VNm 3odl< a a _ a a oNoHNou Nfiooon .a.o oNoHNou NNnoon .a.a :ohooNoo> cowue>wfimm Amecouca Hope we oohsom .m.oooa .N Nooom .4 Hora 4N4Nmoom .m ecu .v.H ucoeHaomxm mo mucocSNm omeaaoo use xfiuovfio on» c“ .:0wua>wfiam mo uczoaw on» cu can .xpwmcoucw macho mo mwcwawn you >o< ouaumaom .nonv ofinmh A.4. Experiment I, Part 4. Salivary Flow Rates to Food Stimuli in the Elderly and College Students. Figure 10 illustrates a comparison of the whole mouth salivary flow rates to sniffing lemons and tasting lemonade between the elderly and college students. Salivary resting levels were lower in the elderly than the college students, but not significantly 50. Of greater interest was the increased salivary flow rates to the lemons by the college students, whereas the elderly demonstrated only a slight increase. Likewise, the elderly averaged only 80% of the flow rate to lemonade as compared to the college students. The nested AOV (Table 31) reveals that, while the judges and treatments differed significantly, the overall group mean and the replication did not. The interaction of greatest importance, the A X C or group by treatment interaction, was significantly different, indicating that the elderly did not follow the same pattern of increased salivation to the food stimuli as the college students. The separate AOV (Table 30) compares the variances of each group. Both groups demonstrated significant levels of variation in the concentration and subject factors. The elderly, however, were a bit more uniform across replications than the college students, as evidenced by a non-significant F-ratio. By use of the mean separation techniques of Scheffe’and Least Significant Difference (LSD), one may note from Table (32) that the elderly did not significantly increase their salivation levels when sniffing the lemons, while the college students did so (using the 184 185 LSD method). The Scheffe’test is considered the most stringent mean comparison technique, while the LSD is the least sensitive. A compar- ison and analysis of the two methods is presented with great clarity by Keppel (1973). Both methods were illustrated in the interest of accurately assessing and evaluating the salivation data, although, in this case, the two techniques came to the identicalconclusion. Figure 10. 186 Average whole-mouth salivary secretion to sniffing and tasting lemons in the elderly and college students in Experiment I.5. 187 EENNQ <>_._ amok mo oonaom .m.H acoefiuomxm cw mucovspm ouofiaoo new xfiuoefio any coozaon Nfisewum uoow on money cemua>wfimm on» me >o< eopmoz .nfimu cane? 189 Table (32). Mean separation methods of Scheffé and LSD on the salivation data of the elderly and college students, Experiment I.5. 1 ’ Whole Saliva (gZZ min) Elderly Colle e (using ScheffETI (using Scheffé) Sequence: . Segpence: -1- -4- -2- -3- -4- -1- -2- -3- 1.08 1.11 1.16 2.22 1.21 1.23 1.59 2.77 Using LSD: Using LSD: -l- -4- -2- 43- -4- -l- ' -2- -3- 1.08 1.11 1.16 2.22 1.21 1.23 1.59 2.77 4* 1 1=First resting, 2=Sniffing, 3=Taste, 4=Final resting. - 2 Underlined values within a row do not differ significantly. A.5. Experiment I, Part 5. Nutrient Analysis of the 3-day dietary recalls. College Students Table (33) lists the means, standard deviations, minimum and maximum values, and percent 1974 RDA for the 25 students. Generally, the college subjects met or exceeded the RDA for eleven nutrients: total protein, vitamins A, C, 81, 82 , niacin, iron, calcium, phos- phorus and magnesium. Nutrients falling short of the RDA were total calories, pyridoxine, folacin, vitamin D, and zinc. Copper levels were 1.39 mg/day, somewhat below the suggested level of 2 mg/day. Elderly Subjects Table (34) lists the means, standard deviation, minimum and maximum values, and percent 1974 RDA for the 25 elderly subjects. Eleven nutrients met or exceeded their 1974 RDA's: total kilocalories total protein, vitamins C, 81, 82, niacin, 812, and A, iron, calcium and phosphorus. Five nutrients which failed to reach the 100 percent- ile of the RDA were 85, folacin, vitamin 0, magnesium and zinc. Total zinc, (10.35 mg), although substantially less than the recommended allowance of 15 mg, was marginally higher than the students' diets. Only two nutrients differed between the two groups: total calories was a bit lower in the college students (86.99 vs. 102.3% RDA) and magnesium levels were a bit lower for the elderly (87.23 vs. 100.34% RDA). 190 191 Table (33). Nutrient analysis of the 3-day dietary recall of the college students, Experiment I. 1’2 COLLEGE STUDENTS _ 1974 Nutrient i 3.0. Min. Max. % RDA Units Energy 2142.04 885.24 765.26 4255.65 86.99 kcal Protein 83.66 39.62 27.54 193.92 159.65 g Fat 82.12 39.82 21.11 183.99 --- g Carb. 268.69 109.11 97.25 526.75 --- g Cholesterol 311.15 180.87 59.02 822.48 --- mg Crude fiber 4431.62 2325.12 918.44 10441.84 --- mg Vitamin C 172.87 103.49 10.84 436.35 384.14 mm Thiamin 1.43 0.83 0.33 '3.97 110.97 m2 Niacin 21.55 11.56 5.31 58.84 134.32 m2 Riboflavin 2.07 1.32 0.57 5.79 135.69 m2 Pyridoxine 1.43 0.89 0.47 4.55 71.47 mg Vitamin 812 3.39 2.55 0.42 12.59 112.98 pg Folacin 300.35 153.99 23.21 740.39: 75.08 pg Vitamin A 9439.03 8784.33 2257.49 33795.96 213.27 U Vitamin D 121.10 142.99 0.21 628.94 30.27 10 Iron 14.02 7.21 5.40 40.16 114.43 mg Calcium 1037.48 670.78 237.08 2919.94 129.68 mg Phosphorus 1395.73 688.16 397.16 3167.77 174.46 mg Sodium 3084.10 1349.38 631.85 5912.21 --- mg Potassium 3344.07 1456.91 1399.71 7609.21 --- mg Magnesium 327.49 218.51 86.05 1050.72 100.34 mg Copper 1.387 0.711 0.453 2.729 69.35 mg Zinc 9.676 6.29 3.21 32.98 64.50 mg Sugars 132.44 62.31 47.49 292.14 --- g 1i? mean, 8.0.: standard deviation; Min.: value. 2 Value fer cOpper (2 mg) is considered reasonable. minimum value, Max: maximum 192 Table (34). Nutrient analysis of the 3-day dietary record of the elderly, ExPeriment I. 1:2 ELDERLY 1974 Nutrient x S.D. Min. Max 8RDA Units Energy 1923.55 272.22 1489.51 2406.82 102.29 kcal Protein 80.42 16.23 54.39 113.96 167.01 g Fat 73.31 15.15 51.45 111.04 ---- g Carb. 247.06 41.02 185.72 320.37 ---- g Cholesterol 367.01 150.47 135.85 687.48 ---- mg Crude fiber 4124.23 816.6 2745.7 6003.2 ---- mg Vitamin C 115.97 31.33 60.0 183.53 257.7 mg Thiamin 1.27 0.33 0.830 2.200 118.4 mg Niacin 20.84 5.32 11.99 32.12 154.80 mg Riboflavin 2.65 0.976 1.01 4.42 215.66 mg Pyridoxine 1.67 0.52 0.87 3.12 83.63 mg Vitamin 812 15.54 12.57 1.76 31.33 517.94. pg Folacin 277.65 91.98 185.35 602.84 69.41 pg Vitamin A 16648.77 9895.38 4136.15 31474.16 409.44 IU Vitamin D 217.27 134.47 18.72 506.92 54.31 IU Iron 16.63 4.17 9.51 27.01 166.30 MG Calcium 855.75 314.2 315.73 1474.52 106.96 MG Phosphorus 1341.91 300.18 808.92 1834.27 167.74 mg Sodium 2407.1 487.39 1488.15 3466.55 ---- mg Potassium 3266.67 550.65 2143.99 4081.78 —--- mg Magnesium 264.94 47.55 2261.26 355.27 87.23 mg Copper 1.884 0.698 0.630 3.19 94.00 mg Zinc 10.35 2.36 6.65 14.81 68.99 mg Sugars 141.07 33.60 85.32 196.42 ---- g 1i: mean, S.D.: value. standard deviation; Min.: minimum value, Max: maximum 2 Value for c0pper (2 mg) is considered reasonable. A.6. Experiment I, Part 6. Hair Analyses for Zinc/Copper in the Elderly and College Students. Table (348) contains the results of the hair analyses for zinc/copper in all subjects participating in Experiment I, II. The college students averaged 214.5:61 ug/g of zinc and 39.2219 ug copper per gram of hair; while the elderly's values were somewhat lower with 188.3r47 ug/g of hair for zinc and 27.5:12 ug/g copper. The zinc/copper values for one elderly subject were lost due to a hand-eye coordination error by this author, allowing the sample to empty onto the floor. Statistical correlations were attempted between the hair concentrations of zinc and copper for both the university students and elderly classifications. The correlation between copper (x) and zinc (y) levels in the hair of university students was: r=.02, while the similar correlation for the elderly group was: r=.58.* The significant positive correlation of the concentrations of zinc and copper in hair for the elderly signifies that higher levels of zinc are usually associated with higher levels of copper. It is interesting to note that this relationship did not exist in the younger university age population. Analysis of variance revealed NSD between the elderly and college in hair zinc (F=2.79), but significant in the c0pper levels at p.05 (F=6.92). 193 194 Table (348). Zinc/copper analyses of hair samples from the college-age and elderly subjects, Experiment I and II. College Elderly Subject Zn Cu Zn Cu 1 212 22 ug/g -1 187 13 2 241 27 175 37 3 310 64 201 41 4 170 S7 209 19 5 202 40 117 23 6 187 45 187 29 7 117 97 219 37 8 199 46 330 61 9 232 29 163 19 10 404 31 147 13 11 312 71 113 19 12 208 41 193 27 13 117 52 214 33 14 151 17 253 29 15 198 33 188 47 16 183 18 137 23 17 179 37 241 19 18 246 41 214 30 19 196 27 * * 20 237 40 163 22 21 246 46 159 27 22 163 22 . 140 19 23 197 19 214 17 24 213 23 193 41 25 242 36 161 14 214.5161 39.2119 188.3147 27.5112 1All values in micrograms/gram hair sample. *Data lost in analyses. 8. Experiment 11, Parts A and B. Odor identification in the elderly, with (Part A) and without (Part B) a list. Figure 11 compares the percent correct response to odor identi- fication of eight food odorants and two blanks by 25 elderly subjects. In Experiment 11, two of the odorants were the same (grape and pepper) while the other six odorants were common household foodstuffs. The results of Experiment 11 were similar to the odor recognition studies of Experiment I. Elderly subjects averaged 22% and 30%, without and with the list, in contrast to the 75% and 90% recognition perfor- mance of the college students. Surprisingly, the elderly increased their recognition scores only 8% above the without list trial. And, if one discounts 6% due to chance guessing alone, the improvement of 2% seems incredibly small. Garlic powder recognition scores actually decreased when subjects used the list (20% to 27%). In contrast, with the aid of the list of food odorants, the college students were able to increase their overall scores by 15% to a 90% level (Figure 12). Lemon and pepper received 100% recognition in the college students, while the best recognized food odors for the elderly were the same, but with 42% recognition values. The least volatile food odor, maple syrup, was the most difficult to recognize for both groups. 195 196 .m .<.11 oeoe_toaxm .6 xpcmupm on» an .mxcopn N was mucocouo voom m 4o cowuooww_u:mup cove on» o» mmcoamoc uumccoo “cause; .11 orsmsu 197 mzz< FITI1IYIrTIITrTTIIT 2 m=¢>m N.E(z ”3030.; 03920 No.2. wa&..( -22a¢mn=m n20_b< wen... w..aa( 'Tulrlti'l'ial'lw' mango.“ .2215. rll 1' ' o—JC‘O """""""""" ' ' ' """""""""""""""""" a=¢>m waa<2 wpm< Cmaawa mxz4”;ko \V\\\\ 8n n z. mom—doc HHU H28.28....»55. cooo ESNOdSBH 1038800 .LNSOHSd 203 .m.- ucmswcmaxm .muummmam omo__oo oso sproo_o oso so .84., 4:3 no.3 .mwcoum cowuoovewucmuw coco one eo comvcoasoo .41 otsmau 204 mx2< c. an an :9 cm on as as am . _ \ ...... r m .1 . m . .. m 1 .l l w L T l r. use“ NEH. I T 5.22 3...? 1 rnl l r upmf. L .. E L 1 m: I. x25... 02:9 (flak L 55266 .. l uuuuuuuuu Letitialuliiunnni 111111111 .L IIIIIII 0356... music cwoomo 2623 3.23... EN H z. >amwoaw V\.\\\... .8 u z. momjoo U c- an ow om cc 8 8. me< a=¢>w wan—<2 mozz. 3.24.. - 3...: a cwaawa 'I'WITTnI'TTI'T‘I' amazon UZC<6 fl 9234. Nim: 501:); rflm whm mum—.25. HZO_._.€U.H=._.an: mOoO OOOOOOOOO mchmmenwp as'ivoasau 1338800 inaouad 212 .m.11 Newspcmaxm 4o mucousum oomppoo apogee use opus asp caravan .umwp o saw: mucoum copuougmvpcmew coco we» mo comwceaeou .81 otamau 213 9.25m N 11211 $25.80 coo... .1 ac mmoom uo< @— on on an cc 2. on on No.2. mama: 39:. fi 2:3» 12.. «23 0.: ll Iwmcco cuaawa .... I I J.M.-1.0.3. lllllllllllllll z 083 1.1.23". .wwwafi , a. u 21 11.1521... E 18.23.. N15: 1:2; 31 .. z. .352 GO NP CG '0 [111111111 L .1 . L c1838 1.038800 1.0130838 J c a NO 1111 69 On p 38 “Wm—(Em... .21 mu...< . j . 4. 1 11. .. L t 1.22... 1 8 . L >Om 1 ... «up 1 8 1 some ._ I 888 <88 59. 1 2. . 29,2226 1 H. 5.1.3.. 15256 L 8 I l 8 . ZO-ZO WA.6.hm.._ zwh up 0002 2:825 Doom 3 .o w6<¢w>< ] 1 1 o r. 1 2 4 1 d 1- |... ON a . . 8 1. 4.. 81% -s m .4 .1 N T J 3 I. t 4 o .. 8.1.18 4 8 W 4 . 8 3 .. r 1 1 s o 4 WWW" cuoauo 82:6 . 1. 1 1 2 . 20232.0 228 . we. I <88 1 8 1.” . 4 O I. 4 J 8 N . 20.20 :OEN .22 \\\S . 11“.. ‘8’ ........................ J 8? ) N BEN 26.8 U _. 8 .m: 5.3\20_._.<0_u=h2wo_ c000 1>._cma._m 220 Table (40). Means of the correct percent response to the odor identification of Experiment III.1, with and without a list, in eight elderly subjects, before and after supplementation with Zn/Cu. e a Without a List1 With a List2 Food ‘ Before3 After 4 Before ‘ After Soy Sauce 17 0 25 13 Pepper 46 50 63 SO Grape 33 63 63 38 Cocoa 29 38 75 63 Tea 25 O 25 25 Root Beer 21 33 63 25 Coffee 38 25 50 SO Cinnamon 38 38 63 63 Orange 17 SO 38 63 Onion 63 63 88 75 i: (mean) 33 36 55 47 1Student's t-test: -0.55 (not significant) 2Student's t-test: 1.65 (not significant) 3Before supplementation of Zn/Cu 221 Part 2. Odor intensities The similarities of odor recognition carried over to the odor intensities. Figure 19 illustrates that the average intensities of seven food odorants were quite similar, before and after Zn/Cu supplementation. Because the trend for five of the seven foods was for increased odor intensity, the four-way AOV found the difference significant at the P<0.01 level (Table 41). If one pools the non- significant 3-way interactions, as some statisticians suggest, the before/after F-ratio is not significant. In anyevent, the trend for most of the food odors was increased odor intensity after the mineral supplementation. The group by odorant interaction (AXB) was also significant, indicating that the Zn/Cu supplementation affected the way the panelists rated all the odorants. However, this inter- action was low (P<0.05) and would be non-significant in pooled data. Part 3. Salivation to food stimuli Figure 20 represents the effect of Zn/Cu on whole-mouth salivation in the eight elderly. Although the two resting levels and tasting lemonade were comparable, there existed an unexpected drop in the response to sniffing lemons after the mineral tablets. Examination of the AOV (Table 42) reveals significant overall means before and after supplementation, highly significant experimental sequence and subject variability, and a non-significant replication factor. The group by sequence interaction was significant at the P<0.05 level. Inspection of the individual data revealed that one subject nearly doubled her overall salivation rate to food stimuli by participation 222 in the eight week supplementation regimen. Interestingly, the subject mentioned previously to the experimenter that her salivation had increased dramatically to food stimuli or no stimuli at all. Throughout the zinc/copper supplementation period, a log was kept each week concerning the subjective remarks by the elderly to the effect(s) of supplementation on taste and aroma perception. Table (25) in the Appendix catalogues these remarks as the study progressed through the eight week period. Although few comments were made concerning changes in gustatory or olfactory acuity, subject number 6 mentioned an increase in salivation near the completion of the experiment. The A X C interaction (before/after by subject) was highly significant, as three subjects increased their overall secretory rates and five decreased. A cautious interpretation of the data suggests that the observed effects were more random than the statistics indicate, especially in non-pooled AOV's of significant variability. Part 4. Hair Zinc/Copper analyses The zinc/copper analyses of the hair samples from the elderly subjects before and after zinc/copper supplementation is listed in Table (43). After the eight week supplementation program, hair zinc values increased from 210 ug/g to 231 ug/g of hair. This increase, although not statistically significant at P<.05, was, nevertheless, of considerable importance as a trend for significance was noted at P<.10. Copper levels, however, demonstrated a slight, nonsignificant increase at 28 ug/g to 33 ug/g of hair. It should be pointed out that the variance between subjects was considerable. 223 .N.111 Newswaoaxm mo cowuoucosmpaaam connoU\u:1N sea cmuem new mcoema .mucocouo uoow =o>mm o» xpcoUFm agave on» »n aavmcoucv gave .11 oesm1u .l It‘ll} 224 322.2% >¢Ohowm ocpflz 2:825 noon. 5 .o 5.23:: w6<¢m>< \ 4 W 33 2 .. m .4 1 “ fl 1 A L 8 . \ 1 1 .4 $ 1 L L ow . ‘ . <1: ...8 .1 .. <88 812m 8 .... .r ammo >8 8 poo: 1 11.28 cmaama 88.8 2 U s N. 82822 VAN 8 1811111.. .. .... .. .... 1 1.. 111111111918. .10ch 2200 I >._..m2m._.z_ mono gamma—4w 173 = N) A.LISN3LNI 8000 30V83AV ('Sd38 8 225 .11os1uooeooe .1eo.o eea .1o.e .mo.ove so oeao111eu1m .... ... .. .mEONHmdsono cwuannm Ham How Houmcweocou ma.voms as x u x m x mo OUHSOW .N.HHH Hfivfiwhomxm .mughovo voom =0>om mo kuwmcvunm eoeo oeo eo 1eooea oee anemone =U\eN mo uoommo on» we >o< xas-uaoa .1141 o1oee Figure 20. 226 Whole-mouth saIivation by the eight eIderly to sniffing and tasting lemonade, before and after the zinc/copper supplementation of Experiment III. 227 d d «I q Rest M r A.5.: «\3 <>.._1oooamon .1oo.o cam 1o.c .mo.cvm um Hemo1m1eu1m .tII ... .. .EHOH HOMH” 0&9 fiQma mOMHdHIQ HH< ”QHOZ mm.~1 we a x u x m x < om.c m~.c mo.11 me a x u x m ..vn.~ mm.o 1m.11 Q1 a x u x < Nm.1 1¢.c me.~ o o x m x < mm.1 mv.o 1N.¢1 11 u x m x < «mtmm.m mo.H Hm.mH Va D x U wo.o 1~.° m~.1 o a x a 1m.o om.o 1e.o N a x < ...me.» 1c.1 m¢.- 1N u x m .**m~.¢1 me.v «c.1m 1 u x < *01.n mm.c mm.~ n m x < mm.o m1.c om.o N m:O1umo11mom -a ...mn.oh 1m.1~ mo.~m1 h mouonnsm -u .t.hm.~1 mm.- ”n.1o n oocozcom -1 .mo.v m«.1 m¢.1 1 =U\=N pouma\oaomam -< o1umn-m .m.z .m.m .m.a :o1oa11a> mo oohsom .U.HHH acoEwnonxm .cofiumuaoEOHQQ9m :0\:N noumw can onomon .xgnovgo pamwo up ovmcoaoa can mcosod wcfiumau and ucwmmficm cu m~o>01 :oHum>flHam ecu mo >o< Amzunsom .mmvv canoe 229 Table (43). Zinc/copper analyses of hair samples from the elderly subjects, befbre and after zinc/copper supplementation in Experiment III. Before/After Before/After Subject (Zinc)2 (Copper)3 8 330/400 ug/gl 61/43 11 113/137 19/23 12 193/240 27/33 13 214/210 33/57 17 241/235 19/24 18 214/240 30/30 23 214/201 17/19 25 161/191 ‘ 14/31 210:63/223176 28i15/33i12 1All values in micrograms/gram hair sample. 2t-test: before/after zinc =-2.16 (not significant) 3t-test: before/after copper = -1.14 (not significant) REFERENCES Hewlett-Packard. "Statistics manual for use with the HP-SS", 1974. Keppel, G. "Design and analysis, a researcher's handbook". Prentice-Hall, New Jersey, 1973. Roessler, E.B. "Applied statistics in the agricultural sciences". Syllabus for Ag Stats. 150, UC Davis, September, 1975. Schiffman, S. Food recognition by the elderly. J. Gerontology, 32, 586-592, 1977. 230 V. DISCUSSION A. STUDE I. Sensory Scaling, Salivary Flow Rates and Odor Recognition between Twenty-Five College Students and Twenty-Five Elderly Subjects. Sensory Scaling In the analysis of the hedonic responses to varied levels of NaCl in tomato juice or to the sucrose levels in lemonade, one must remember the great variability of response inherent in hedonic judgments. As was pointed out by Trent and Pangborn (1981), a significant statistical difference in hedonic scores may obscure the correct interpretation of individual variance. In the tomato juice experiment, although the elderly and college students judged the overall degree of liking for the samples as being significantly different, inspection of the individual graphics reveals an amazing similarity in response. while elderly possessed a slightly restricted range of response, only a few individuals had difficulty in the sensory discrimination of the samples. A similar situation existed for the lemonade hedonic scores--the two groups were significantly different in overall hedonic scores but inspection of the individual graphics revealed remarkable similarity in direction and magnitude. Similarly, truncation of the intensity respbnses to NaCl levels in tomato juice and sucrose in lemonade was noted in the elderly. Inspection of the individual data, however, found that two subjects possessed flat sensory functions for both the hedonic and intensity 231 232 scalings. In the only other study on age and suprathreshold sensory scaling, Enns g£_al, (1979) concluded that old age does not produce a noticeable deficit in suprathreshold sensory scaling. It must be pointed out, however, that the mean age of their subject population was 71 years as compared with the 83 years of age in Experiment I. The failure to find moderate gustatory deficits in suprathreshold scaling could not have been predicted with the literature on discrimin- ation of threshold testing. A great majority of the sensory studies found taste impairment in the elderly when threshold measurements were taken (Hinchcliff, 1958; Richter and Campbell, 1940; Cooper gt 31,, 1959; Hermel gt_al,, 1970; Balogh and Lelkes, 1961; Cohen and Gitman, 1959; Greger and Geissler, 1978; Langan and Yearick, 1976). Grzegorczyk gt_al, (1979), however, criticized many of these early threshold studies by virtue of their poor methodologies, inadequate subject numbers, or contradictory conclusions. Grzegorczyk gt_al,, in their threshold data, concluded that the elderly possess only a slight impairment in threshold abilities. In the light of the recent study by Grzegorczyk gt_al, (1979) and the suprathreshold study by Enns gt_al, (1979), one may conclude that while the elderly may have a slight to moderate impairment in absolute or detection thresholds to one or more of the four basic tastes, suprathreshold measurements are less affected than one would predict from the myriad of literature threshold studies. 233 Olfaction In contrast to the sensory studies on gustation which demon- strated a remarkable similarity of response between the elderly and college students, the results of the olfactory studies--where the elderly, in general, averaged a percent correct response one-third that of the college students--were quite unexpected. In fact, every effort was made to placate the concerned panel members when they were unable to detect the aroma in any of the ten samples. Quite often the panelists commented on their lack of olfactory acuity with such remarks as "my sniffer isn't working today", and "I haven't been able to smell (foods) for a number of years now". A review of the individual aroma identification revealed that about one-half of the elderly (12/25) possess essentially no ability to detect food aromas as presented. Even foods with a high degree of volatility (eg., onions or pepper), failed to elicit an increase in recognition. Although many of the panelists did not identify the onion and pepper samples, a few subjects did comment on a tingling sensation present in the nasal cavity. It was as if the "quality" of the odorant was absent, while the trigeminal or "pain“ component continued to reSpond, however weak. Although olfactory sensitivity testing has not received as much research as gustatory discrimination, the few studies that were completed demonstrate a surprising concordance with the present study. Chalke and Dewhurt (1957), Kimbrell and Furchtgott (1963) and Venstrom and Amoore (1968) all agree that olfactory acuity in the aged is decreased compared to younger age controls (subjeCts generally under 40 years of age). Schiffman (1979) in a study on olfactory sensitivity in the 234 elderly demonstrated that the elderly (mean age 73.3 years) had moderate difficulty in identifying five fruit aromas; i.e., cherry, grape, lemon, orange and tomato. In comparing Schiffman's study with the present work, it must be noted that her subjects tasted pureed food samples while blindfolded, which contrasts with the straight olfactory testing here. The additional recognition cue of tasting the pureed food complicates a comparison of the data. Therefore, it is not surprising that Schiffman's subjects were able to identify the foods with a greater degree of accuracy than in Experiment I. Schiffman (1979), in the discussion on olfaction and aging, speculated that memory impairment may be a significant factor in the decreased recognition scores for food odorants. The design and out- come of Experiment 1.3, part 2, however, must cast some doubt on this conclusion. The addition of a "list" of the odorants in the sensory testing booth increased the accuracy of the elderly's reSponse only 20% over the previous level. An intriguing question arose during the intensity testing of the seven food odorants by the elderly. Is it possible to perceive an odorant without being able to identify it? The data of Experiment I suggests an affirmative answer. As mentioned previously, 12/25 subjects possessed essentially no ability to recognize food odorants; however, during the food aroma intensity rating sessions, these same subjects perceived a food aroma and rated it as such. Although this experimenter cautioned the subjects to give a zero value to aromas not perceived, subjects consistently gave intensity ratings to foods they could not identify. Although there is no precedent in the literature demonstrating this effect, one might speculate a 235 presence of a "universal" aroma that might be present and perceived. This nondescript aroma might arise from a failure to integrate lower order intensity perceptions. Schiffman (1979), in a review on the biological aspects of olfaction and aging, suggested that there are age-related structural degradations in the hippocampus and amygdala. If these two areas in the brain are responsible for quality interpre- tation, then the possibility of perceiving a stimulus without discrim- ination is a plausible conclusion. An alternate hypothesis might be that the surface cilia in the olfactory region, through the aging process, have increased detection thresholds to a point where sensory input does not support sensory discrimination.. Schiffman (1979) concluded that: ". . . the loss of taste quality information, i.e., loss of distinctions between stimuli, following neural losses means that all stimuli must taste somewhat more similar to each other than before such losses. In other words, with neural losses various stimuli may tend to elicit a common . . ." By substituting the word "taste" with "olfaction", the same conclusion must be drawn here. Salivation One way to check the validity of the olfactory intensity data by the elderly might be to measure salivation induced by the aroma of foodstuffs. Reflex salivation is known to be a fairly linear process-- higher strengths of odorants induce greater salivary secretions (Pangborn gt_gl, 1979). Therefore, one might speculate that the elderly would not salivate as greatly to the aroma of the lemons as the college students by virtue of their poorer intensity responses to the seven 236 food aromas. The results indicate that the elderly, in general, did not salivate across all treatments to the same level as the college students. In particular, responses to sniffing the lemons and tasting lemonade were depressed moderately. This decrease in salivation in the elderly group agrees with the few published reports in the literature (Meyer gt_al,, 1937; wainwright, 1943; Gutman and Ben Aryeh, 1974; and Makila, 1977). Although not stated explicitly in his article, Chauncey _t_gl, (1980) found little correlation between age and salivary flow rates. The population tested, however, was divided into two groups--above and below 55 years of age. Only 10 of his subjects were above 60 years of age, which may be an insufficient number to delineate differences in salivation when the total group tested was 153 individuals. Nutrient Analysis Calculation of nutrient intakes from the 3-day dietary recalls revealed surprisingly little differences between the elderly and college age subjects. Only total kilo-calories and magnesium levels were found to be lower in the elderly than in the university students. This result is not surprising, considering the fact that the elderly resided in a retirement facility with an excellent food service program. Using the 1974 RDA as a comparison, the college students were somewhat low in kilo-calories, pyridoxine, folacin, vitamin D, copper and zinc. Inspection of the individual dietary records revealed the abnormally low levels of kilo-calories among the thirteen women participants. Eight of the women recorded total calories less than 1200 kcals. The 237 current fashion trend to stay slim through dieting may be responsible for these results. Indeed, White (1976) mentions that diets low in energy values are not unusual for college women. Diets low in kilo- calories usually are low in other nutrients as well (White, 1969), especially the mineral elements. Therefore, it is not unusual, when all the data from the males and females are averaged together, to find the diets low in certain mineral levels such as zinc and copper. As White (1976) and Holden 22.21: (1979) point out, caloric intakes less than 1000 kcals rarely provide 100% of the RDA for zinc or copper. The data on the college students, however, agree with the report by Holden gt_gl, (1979) which found 4.19 mg zn/1000 kcals as compared with 4.5 mg zn/1000 calories in our study. Surprisingly, the elderly, although below the RDA's for pyridoxine, folacin and vitamin D, were a bit higher in their levels of zinc and copper (1.884 and 10.35 mg, respectively) than university students. These values are similar to the values of Deeming and Weber (1978)-- 11.5 mg zn and 1.7 mg cu--and Marshall gt_al, (1975). Hair Analysis In Study 1, the university students averaged 214 mcg of zinc and 39.2 mcg of copper per gram of hair sample, while the elderly's values were somewhat lower with 188.3 mcg of zinc and 27.5 mcg copper per gram of hair. In the discussion of zn/cu levels in hair and the comparison of the two age groups, it is useful to refer back to Table 9 of the dissertation. Generally speaking, the values for zn/cu per gram of hair sample in the university students were a bit higher than most 238 of the studies cited, but well within the range of variance noted. A recent report by Vir and Love (1981) reconfirms this finding. In a study of 24 young women comparing the effects of oral contraceptives on zinc and copper nutriture, the authors found an average zinc content of hair near 175 pg/g and of copper, 13.4 ug/g in the control subjects. Large standard deviations in these studies underscore the need to be cautious in absolute number comparisons. Fewer studies have determined the trace mineral contents of zinc/copper in the elderly. Vir and Love (1979) studied the zinc and copper status of the elderly with ages ranging from 65 to 95 years. These subjects were studied under two groups, with and without multi- vitamin supplementation. The control groups showed a mean zinc value of 208 ug/g hair and a mean copper level of 12.0 pg/g hair. It is interesting to note that the range of zinc values for the elderly group was 85-490 pg zn/g hair. In the multivitamin group (no zinc or copper added) mean zinc values increased to 245 pg/g hair and copper to 15.8 pg/g hair. The apparent association between the increase in vitamin intake and plasma or hair level of cations was not explained by the authors. However, it does serve as a caveat to investigators on the variety of factors that might influence trace metal nutriture. As far as the influence of age on zinc and copper contents of hair is concerned, the literature is markedly equivocal. Kelvay has stated that both copper and zinc levels in hair increase with the age of the subject (Klevay, 1970a; Klevay, 1970b). Deeming and Weber (1978), however, stated that not only do hair minerals correlate poorly with serum levels or dietary intake, hair zinc decreased with age. Petering (1971), however, demonstrated that female subjects increase 239 their hair copper with age but hair zinc decreases. Males, on the other hand, show a decrease in both hair zinc and copper with age. To further confuse the issue, Hambidge (1973) emphatically states that interpretation of analytical data on hair copper concentrations requires great caution. Analyses, the author states, should be limited to recently grown hair within 1 t012 cm of the scalp. It is obvious that until a more satisfactory method of measuring trace elements in hair appears (e.g., erythrocytic zinc, Robson and Spell, 1981), the researcher must exercise extreme caution in the preparation of samples and the interpretation of data in all hair biopsy studies. B. EXPERIMENT II. Odor Identifications in the Elderly and College Students. The research intentions of Experiment 11 were twofold. First, to discern whether the elderly subjects would improve their odor recogni- tion scores with practice or with a list of foods placed in the booth; and secondly, to enumerate what classes of foods, if any, the elderly might assign to the blank sampling cups. In Experiment II, five new foods were added to the list, two of the odorants were retained from the first experiment and the two blank cups were randomly placed with the others. _ As in Experiment I, the elderly experienced great difficulty in odor recognition, even with the list of foods appearing in the sensory booth. The two food items which the elderly firSt encountered in Experiment I received similarly poor recognition scores. If experience were a factor in aroma identification, one might expect a higher correct reSponse than the results indicated. The college students, in marked contrast, were able to approach 100% recognition for all but the difficult-to-identify maple syrup. Of considerable interest was the finding that the elderly were able to identify the blank sample 50% of the time. This result suggests that the elderly, while having great difficulty with aroma discrimination, could still identify an empty sample cup. Again we see support for the theoretical "universal" aroma of the elderly--a phenomenon where the quality of an aroma is not discernable but the perception of an undefinable stimulus is. Although Schiffman (1977) commented in her paper on food recogni- tion and the elderly that the decline in smell sensitivity is greater 240 241 than the decline in taste sensitivity, the presence of a universal aroma was not postulated as an explanation for sensory decline. The grouping of misnamed food aromas by the elderly did not form any discernible trend or classification. Curiously, Schiffman (1979) found that elderly subjects were better at guessing fruit aromas over any other categories, i.e. meat, milk, vegetables, etc. In Experiment II, the elderly were encouraged to verbalize all guessing; however, the natural tendency of these subjects to be conservative in their opinions may have masked the proper interpretation of our results. C.' EXPERIMENT III. Zinc/Copper Supplementation of Eight Elderly Subjects. Although several dozen studies in the literature examined the use of zinc/copper to restore or treat taste and smell dysfunction, (Henkin, gt_al,, 1969), only one study attempted to improve taste acuity in the elderly. In this study, Greger and Geissler (1978) found that 15 mg of zinc for 95 days essentially unaffected the taste acuity thresholds for the four basic tastes. In our study, although copper was added and the zinc dosage was higher, the findings echo those of the 1978 study by Greger and Geissler. Odor recognition, aroma intensities and salivation to food stimuli were essentially unchanged. In fact, some of the data on odor identification and intensity suggest a decrease in response after the zinc/copper supplementation. These artifacts of statistical data do little except confuse the interpreta- tion of the supplementation results. The salivation data are of even greater variability. Although the decrease in salivation to food stimuli is not significantly different, this curious response remains unexplained at present. Several possible explanations might exist to explain the lack of response to zinc/copper therapy. First, and most obvious, is the confusion surrounding the link between trace metal ingestion and sensory restoration (Catalanotto, 1978). This author concluded that zinc depletion can lead to decreased taste acuity, but decreased taste acuity is not necessarily associated with depletion of zinc. Since both the dietary recalls and hair mineral analysis of the elderly in Experiment I do not suggest a depleted zinc/copper body state, it is not 242 243 surprising that additional Zn/Cu in the diets failed to affect sensory acuity. A second explanation concerns that actual amount of Zn/Cu absorbed from the supplement by the elderly. The hair analyses of the elderly after supplementation demonstrated only a moderate increase in Zn/Cu after eight weeks of mineral ingestion. Perhaps the levels of added Zn/Cu were insufficient to increase body stores to a level where sensory acuity is affected. Aamodt gt_§1, (1981) demonstrated that human zinc absorption is age dependent. In their study, 75 volunteers aged 18-84 were studied for zinc absorption using total body counting of Zn 65. The authors concluded that zinc absorption decreases linearly with age in both men and women. In another study, Taper gt_gl, (1980) added supplements and ZnSOu and CuSOu to a three day cycle menu in ten subjects aged 68.1 years. The authors noted a great variability in the retention of zinc in their patients, with the level of zinc intake (7.8 or 23.3 mg/day) having no effect on zinc retention. Both of the studies cited above demonstrate the difficulty in increasing body stores of Zn/Cu, whether used in dietary intake or absorption studies. In fact, Friedman gt_gl, (1980) found that hair zinc and salivary sediment zinc levels did not increase after 15, 50 and 100 mg zinc. Perhaps the form of the supplement used is of greater importance than the amount. In any event, additional research on Zn/Cu absorption and retention is needed to discern the most efficient method of increasing the mineral stores of the body. VI. GENERAL CONCLUSIONS Gustation The overall results of the taste responses to sucrose in lemonade and salt in tomato juice suggested a great similarity in response between the college students and the elderly. Although there was evidence of a flattened psychophysical function in the overall response, the suprathreshold responses were not nearly as depressed as one would expect from a prior knowledge of the threshold studies in the literature. Olfaction Quite unanticipated were the results of the olfactory studies demonstrating a highly depressed level of olfactory acuity and recognition exhibited by the elderly. Although the literature contains several reports of decreased olfaction in older subjects, the extent of the decline was hardly anticipated. The anecdotal accounts of the little old lady who applies too much perfume in grooming for church activities is now partially explained. As to the etiology of athis condition of anosmia, whether partial or complete, one must wait further investigation. Why olfaction is more affected than gustation remains another question subject to future scientific inquiry. While Schiffman (1980) has speculated on the finding of atrophy in certain brain structures relating to olfaction, why the sense of taste is not as profoundly affected remains unclear. 244 245 Zinc/Copper and Taste Although the attempt to restore "normal" olfactory acuity in the elderly with the addition of Zn/Cu to the diets was unsuccessful, the results, however negative, shed light on the complexity of linking mineral nutrition with a psychophysical task. Aging is a phenomenon of complex organ and subcellular changes over time. The likelihood of reversing any one of these changes with a simple change of diet must be considered a formidable task at best. Simple solutions, while therapeutically and experimentally enticing, have yet to be elucidated by this or anyone, else's research protocols. Perhaps the etiology of decreased sensory acuity with age is a simple one. The application of this knowledge, however, to reverse or delay this impending decline is likely to be a challenging task, not only for psychophysicists involved in its resolution, but also medical personnel involved with its application. 10. 11. REFERENCES Aamodt, R.L., Rumble, W.F., Johnston, G.J., Markley, E.J. and Henkin, R.I. Human Zinc absorption is age dependent. Fed. Proc., 40, 950, 1981. Balogh, K. and Lelkes, K. The tongue in old age. Gerontol. Clin., 3, 38-54, 1961. Catalanotto, F.A. The trace metal zinc and taste. J. Am. Clin. Nutr., 31, 1098-1103, 1978. Chalke, H.D. and Dewhurst, J.R. Coal gas poisoning: loss of sense of smell as a possible contributory factor with old people. Br. Med. J., 2, 1915-1917, 1957. Chauncey, H.H., Wallace, 5. and Alman, J.E. Salivary Chloride Levels, Taste Thresholds for Salt, and Food Ingestion. In: "Biological and Behavioral Aspects of Salt Intake", Academic Press, pp. 113-125, 1980. Cooper, R.M., Bilash, I. and Zubek, J.P. The effect of age on taste sensitivity. J. Gerontol., 14, 294-298, 1959. Deeming, 5.8. and Weber, G.W. Hair analysis of trace minerals in human subjects as influenced by age, sex, and contraceptive drugs . J. Am. Clin. Nutr., 31, 1175-1180, 1978. Enns, M.P., Van Itallie, T.B. and Grinker, J.A. Contributions of age, sex and degree of fatness on preference and magnitude estimations for sucrose in humans. Physiol. Behav., 22, 999-1003, 1979. Greger, J.L. and Geissler, A.H. Effect of zinc supplementation on taste acuity of the aged. J. Am. Clin. Nutr., 31, 633-637, 1978. Grzegorczyk, P.B., Jones, S.W. and Mistretta, C.M. Age-related differences in salt taste acuity. J. Gerontology, 34, 834-840, 1979. Gutman, D. and Ben-Aryeh, H. The influence of age on salivary content and rate of flow. Int. J. Oral Surgery, 3, 314-317, 1974. 246 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 247 Hambidge, K.M. Increase in hair copper concentration with increasing distance from the scalp. J. Am. Clin. Nutr., 26, 1212-1215, 1973. Henkin, R.I., Graziadei, P.O.G. and Gradley, D.F. The molecular basis of taste and its disorders. Ann. Intern. Med., 71, 791-821, 1969. Hermel, J. Schonwetter, S. and Samueloff, 5. Taste sensation and age in man. J. Oral. Med., 25, 39-42, 1970. Hinchcliff, R. Clinical quantitative gustometry. Acta Otolarynggl., 49, 453-466, 1958. Holden, J.M., Wolf, W.R. and Mertz, W. Zinc and copper in self- selected diets. J. Am. Dietet. Assoc., 75, 23-28, 1979. Kimbrell, G. and Furchtgolt, E. The effect of aging on laboratory threshold. J. Geront., 18, 364-365, 1963. Klevay, L.M. Hair as a biopsy material. 1. Assessment of zinc nutriture. J. Am. Clin. Nutr., 23, 284-289, 1970b. Klevay, L.M. Hair as a biopsy material. II. Assessment of copper nutriture. J. Am. Clin. Nutr., 23, 1194-1202, 1970a. Langan, M.J. and Yearick, E.S. The effects of improved oral hygiene on taste perception and nutrition of the elderly. J. Gerontol., 31, 413-418, 1976. Makila, E. Oral health among the inmates of old people's homes. Salivary secretion. Proc. Finn. Dent. Soc., 73, 64-69, 1977. Marshall, M.W., Iacono, J.M., Young, C.W., Washington, V.A., Slover, H.T. and Leaply, P.M. Composition of diets containing 25 and 35 percent calories from fat. J. Am. Dietet. Assoc., 66, 470-481, 1975. Meyer, J., Golden, J.S., Steiner, N. and Necheles, H. The ptyalin content of human saliva in old age. Am. J. of Physiology, 119, GOO-602, 1937. Pangborn, R.M., Witherly, S.A. and Jones, F. Parotid and Whole Mouth secretion in Response to Viewing, Handling and Sniffing Food. Perception, 8, 339-346, 1979. Petering, H.G., Yaeger, D.W. and Witherup, S.D. Trace metal content of hair. I. Zinc and copper content of human hair in relation to age and sex. Arch. Environ. Health, 23, 202-207, 1971. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. 38. 39. 248 Richter, C.P. and Campbell, K.H. Sucrose taste thresholds of rats and humans. Am. J. Physiol., 128, 291-297, 1940. Robson, J.R.K. and Spell, L. Erythrocyte zinc. J. Am. Clin. Nutr., 34, 1983, 1981. Schiffman, S. Food recognition by the elderly. J. Gerontol., 32, 586-592, 1977. Schiffman, S. Changes in taste and smell with age: psycho- physical aspects. In: "Sensory Systems and Communications in the Elderly", Aging, Vol. 10. EDS., Ordy, J.M. and Brizzee, K., pp. 227-246, K. Raven Press, New York, 1979. Schiffman, S.S., Hornack, K. and Reilly, 0. Increased taste thresholds of amino acids with age. J. Am. Clin. Nutr., 32, 1622-1628, 1979. Schiffman, S. Magnitude estimation of amino acids for young and elderly subjects. In: "Olfaction and Taste VII", Edited by H. Van der Starre. IRL Press, London, pp. 379-383, 1980. Taper, J.L., Burke, D.M., DeMicco, F.J. and Ritchey, S.J. Zinc and c0pper interaction and retention in elderly human subjects. Fed. Proc., 39, 430, 1980. Trant, A.5., Pangborn, R.M. and Little, A.C. Potential fallacy of correlating hedonic responses with physical and chemical measurements. J. Food Sci., 46, 583-588, 1981. Venstrom, D. and Amoore, J.E. Olfactory threshold in relation to age, sex and smoking. J. Food Sci., 33, 264-265, 1968. Vir, S.C. and Love, A.H.G. Zinc and copper status of the elderly. J. Am. Clin. Nutr., 32, 1472-1476, 1979. Vir, S.C. and Love, A.H.G. Zinc and copper nutriture of women taking oral contraceptive agents. J. Am. Clin. Nutr., 34, 1479-1483, 1981. Wainwright, W.W. Human saliva XV. Inorganic phosphorus content of resting saliva of 650 healthy individuals. J. Dental Res., 22, 403, 414, 1943. White, H.S. Inorganic elements in weighed diets of girls and young women. J. Am. Dietet. Assoc., 55, 38-43, 1969. White, H.S. Zinc content and the zinc-to-calorie ratio of weighed diets. J. Am. Dietet. Assoc., 68, 243-245, 1976. VII. SUGGESTIONS FOR FURTHER RESEARCH Sensory research involving the influence of aging and the perceptions of gustation and olfaction has received only limited attention by psychophysicists. Taste thresholds--detection, recogni- tion and olfactory--are well characterized for a variety of food substances in a population of younger individuals, generally 50 years or less. What is needed, therefore, is a concerted effort to tabulate all threshold data in the elderly using modern sensory evaluation procedures and principles. Preferably, these studies should include a younger group of subjects tasting or sniffing the identical substances under similar conditions. Already there are indications of renewed interest in elderly taste threshold work as evidenced by the recent work by Schiffman (1979) on the "Increased Taste Thresholds of Amino Acids with Age". Suprathreshold studies are equally important in the assessment of sensory decline during the aging process. As stated in the literature review, only one study compared the sensory scaling of suprathreshold concentrations of tastants between the elderly and a younger population directly (Schiffman, 1980). Additional studies are needed to confirm or to elaborate on the present finding of this dissertation that the elderly possess flattened psychophysical functions for sucrose (sweetness perception) and salt. The other two basic tastes-sourness and bitter perception-have yet to be tested in. an elderly population. 249 250 Additional research on olfaction is necessary to confirm the finding of a "universal" aroma perception in the elderly. Studies based on intensity scaling of a wide variety of food substances should clarify this puzzling phenomenon. A more reliable index of Zn/Cu status than hair analyses is probably necessary before one can correlate dietary intakes and the mineral nutritional state in individuals. Saliva, enzyme assays or ’plasma levels of Zn/Cu are candidates for short term indicators of mineral stores. Correlations between these values and sensory studies of gustation or olfaction may prove doubly rewarding. VIII. APPENDIX No. Page I. Letter to all residents of Burcham Hills encouraging participation ....................... 253 II. Information bulletin given to all residents of Burcham Hills Retirement Center .................. 254 III. Follow-up information bulletin to residents of Burcham Hills ....................... 256 IV. Medical record form used to assess drug intakes in the elderly prior to Experiment I ............... 257 V. Consent form signed by all individuals prior to the participation in Experiment I ................ 258 VI. Scorecard used in the hedonic evaluation of saltiness in tomato juice ...................... 259 VII. Scorecard used in the intensity evaluation of tomato juice saltiness . .‘ ..................... 260 VIII. Scorecard used in the hedonic evaluation of sweetness in lemonade ........................ 261 IX. Scorecard used in the intensity evaluation of lemonade sweetness ......................... 262 X. Composition of tomato juice used in Experiment I ....... 263 XI. Product information on food products used in the odor recognition Experiment 1.3. ................ 264 XII. Scoresheet used in odor recognition and intensity evaluation of Experiment 1.3. ............... 265 XIII. Dietary recording forms used by the college students for their 3-day food records in Experiment I .......... 266 XIV. Actual coding form used to record dietary intakes of subjects in Experiment I .................. 270 XV. Actual food items served during the 3-day dietary recording of the elderly in Experiment I ......... 271 XVI. Basic measurements and ladle equivalents used by the Burcham Hills staff to portion out food items to the elderly, during the 3-day dietary food recall ............ 273 251 No XVII. XVIII. XIX. XX. XXIA. XXIB. XXII. XXIII. XXIV. XXV. XXVI. 252 Product information on f00d products used in the odor recognition, Experiment II Handout distributed to the elderly participants near the end of Experiment II Handout given to all elderly participants of Experiment II, near the end of the olfactory studies ........... Anthropometric data on college students participating in Experiment I, II Anthropometric data on elderly subjects participating in Experiment I, II Anthropometric data on the elderly subjects participating in Experiment III ..................... Comparison of the anthropometric data of college students and elderly subjects in Experiment I ............ Answers to the following questions posed in the medical record survey of the elderly ................ List of drugs mentioned in the medical record of the elderly in Experiment I ‘ Comments by elderly subjects during the course of their zinc/copper supplementation period in Experiment III . Equations by Durnim and Womersley (1974) used to predict % body fat from a triceps skinfold measurement . . . . ... Page . 274 275 276 277 278 279 280 281 282 . 283 287 253 October 15, 1979 TO ALL RESIDENTS OF BURCHAM HILLS: Thank you for your attention during my ten minute presentation of the taste project. I would also like to take this opportunity to thank Mr. George Redheffer for his enthusiastic support of this study. We would like to invite all residents to participate in the study conducted by Dr. R. A. Schemmel and Steven A. Witherly. If you would like to join us in what plans to be an exciting project, we ask that you would read the following materials: 1. INFORMATION BULLETIN: everything about the study is contained right there. 2. CONSENT FORM: if you wish to participate, please read the consent ferm and sign your name and put the apt. number down so that we might be able to contact you. 3. FINALLY: we will need a little information regarding your state of health since this may influence results. This infbrmation will be collected privately in your room. We appreciate your concern in this project. I will be by later to pick up the consent forms and contact each of you individually. Yours truly, Steven A. Withe y Figure 1. Letter to all residents of Burcham Hills encouraging participation. 254 INFORMATION BULLETIN To all interested subjects . We would like to invite all persons residing in: to participate in a unique scientific experiment involving the senses of taste and smell. People willing to join our study will be involved in the fellowing: 1. 6. All subjects will taste tomato juice with various levels of salt and taste lemon drink with varying levels of sugar. We ask you to rate how sweet or salty the samples are and how much you like each sample. Samples are not swallowed-- only tasted! Subjects will be asked to identify and then rate the inten- sity of each food odor. Food samples will be in containers that mask visual cues, but allow one to sniff the ten samples. A small bit of hair is needed from each subject; a snip from the back of the neck will prove adequate. Only two samples are needed: one in the beginning and one at the end of the experiment. Finally, we wish to take salivary samples, collected very simply through the use of three cotton dental rolls (Johnson and Johnson No. 2, 1/2 by 3/8 in.) which are placed in the mouth. Subjects merely place them in the mouth, then sniff lemon wedges and taste lemonade. Saliva soaks into the rolls and by immediately re-weighing them, the amount of saliva produced is determined. At the end of the taste-testing we wish to collect a small amount of saliva by having our panelists drool into a plastic container. When the above data have been compiled, the groups of subjects will be broken up into two groups, with one group receiving a mineral supplement and the other receiving a placebo. Neither the subjects nor the investigators will know who receives what. This prevents bias in the final results. The supplementation period will last 2 months, with a pill taken once a day. Finally, all subjects will be retested as outlined above (1-5). The project will be quite interesting to those of you who have not participated in taste-testing. The study has been designed with your safety in mind, and we do not anticipate any problems. 255 Information Bulletin - cont. THANK YOU FOR LISTENING TO US AND READING THIS BULLETIN. IF ANY QUESTIONS ARISE, FEEL FREE TO CONTACT US ANYTIME AT YOUR LEISURE. Figure II. Information bulletin given to all residents of Burcham Hills. 256 October 26, 1979 TO OUR FRIENDS IN BURCHAM HILLS: A few weeks ago a letter was placed in your mailbox in regards to a taste project to be conducted here. Unfbrtunately, we did not receive as many responses as we had anticipated. In order to generate additional interest in our study and diSpel any fears you may have about it, we are planning to conduct a taste demon- stration to be held in the SNACK BAR on Oct. 30 at 9:30 a.m. By doing so, you will have the Opportunity to see and taste for yourself the procedures to be followed in our project. Please come! Everyone is welcome! Previous experience has shown that taste-testing can be a lot of fun, since most people that we have tested said that they hated to see it end. Even if you feel you are unable to taste or have a low level of taste ability, we are interested in having you as a part of our project. Thank you very much fer your time. If interested, please check the "YES" box below and return this portion to Jane Greene by Oct. 29. If not interested, you are more than welcome to stop by and SEE a taste session in progress. Perhaps you will change your mind. Sincerely, Steven A. Witherly, Project Director Michael D. Dauria, Project Coordin- ator PLEASE DETACH HERE AND RETURN THIS PORTION TO JANE GREENE YOUR NAME: YES, I would be interested in participating in your project. NOTE: If you are one of the few who did respond earlier, please ignore this letter. We will contact you directly. Figure III. Follow-up information bulletin to residents of Burcham Hills 257 MEDICAL RECORD: ELDERLY 1. Are you taking any drugs? 2. Are you taking any vitamins? 3. Are you taking any other pills? 4. Do you smoke? 5. Do you have any of these illnesses? Diabetes? Kidney Disease? High/Low Blood Pressure? 6. Is there anything else we should know concerning your medical history? Figure IV. Medical record form used to assess drug intakes in the elderly prior to Experiment I. 258 Consent form for participation in study Michigan State University Dept. of Food Science 8 Human Nutrition I have freely consented to take part in a scientific study being conducted by: under the supervision of: Academic Title: The study has been explained to me and I understand the explanation that has been given and what my participation will involve. I understand that I am free to discontinue my participation in the study at any time without penalty. I understand that the results of the study will be treated in strict confidence and that I will remain anonymous. Within these restrictions, results of the study will be made available to me at my request. I understand that my participation in the study does not guarantee any beneficial results to me. Signed Date Figure V. Consent form signed by all individuals prior to the participation in Experiment I. 259 DEGREE OF LIKING FOR TOMATO JUICE JUDGE: SESSION: DATE: INSTRUCTIONS: Taste samples in order presented. Place a relatively large volume of tomato juice in the mouth, move it around with the tongue, swallow a small amount and expectorate the remainder. Indicate how much you like each sample by circling one of the horizontal marks on the vertical line. Between each sample, rinse the mouth with distilled water. like extremely like extremely dislike extremely dislike extremely Figure VI. Scorecard used in the hedonic evaluation of saltiness in tomato juice. 260 INTENSITY OF SALTINESS IN TOMATO JUICE JUDGE: SESSION: DATE: INSTRUCTIONS: Taste samples in order presented. Place a relatively large volume of tomato juice in the mouth, move it around with the tongue, swallow a small amount and expectorate the remainder. Indicate how much you like each sample by circling one of the horizontal marks on the vertical line. Between each sample, rinse the mouth with distilled water. extremely salty extremely salty no saltiness no saltiness Figure VII. Scorecard used in the intensity evaluation of tomato juice saltiness. 261 DEGREE OF LIKING FOR LEMONADE DRINK JUDGE: SESSION: DATE: INSTRUCTIONS: Taste samples in order presented. Place a relatively large volume of lemonade in the mouth, move it around with the tongue, swallow a small amount and expectorate the remainder. Indicate how much you like each sample by circling one of the horizontal marks on the vertical line. Between each sample, rinse the mouth with distilled water. like extremely like extremely dislike extremely dislike extremely Figure VIII. Scorecard used in the hedonic evalutation of sweetness in lemonade. 262 INTENSITY OF SWEETNESS IN LEMONADE DRINK JUDGE: SESSION: DATE: INSTRUCTIONS: Taste samples in order presented. Place a relatively large volume of lemonade drink in the mouth, move it around with the tongue, swallow a small amount and expectorate the remainder. Indicate how much you like each sample by circling one of the horizontal marks on the vertical line. Between each sample, rinse the mouth with distilled water. extremely sweet extremely sweet T10 sweetness no Sweetness Figure IX. Scorecard used in the intensity evaluation of lemonade sweetness. 263 Table X. Composition of tomato juice used in Experiment 1. Tomato Juice: Featherweight Brand1 NUTRITION INFORMATION Calories ............... 35 Fat ............... 0 Protein ............... 2 g Sodium: not more than Carbohydrates ........... 7 g 10 mgs. in 100 g. Percent of U.S. recommended daily allowance (U.S. RDA) Protein 2 Niacin 8 Vitamin A : 20 Calcium 2 Vitamin C : 2 Iron 4 Thiamin . 4 PhOSphorus: 2 Riboflavin: 2 Magnesium : 4 1Packed for Chicago Dietetic Supplies, Inc., La Grange, IL, 60525, U.S.A. 264 .moec1 .>z .me1e1e eo1ez .meooa 1eeoeou .ee1em 0:1; .emoom .e1m HmAUMMMHAe .ouecwomwo ESMoOm .ufiem .omo131uoo 1xcuosxxonaeo .1fio ofinmuowo> oo~m=uwonv>s xflaewuhen .mowuom mahxm :Hoo .wouou .xufia awn Hammo: .mesm .Ncmme .o1eo .e1oeeoe< .muosoonm coon zonu ea .uoHoo aoeenmo ace ashxm :Aoo .uHem .memon xom seam muoehuxo aflououm .Aouez .Hommv .ofiao .wumenwoewu .00 women: .coamczuo ocsonm chad .wommom xoean ocean» chad .Ncmam .n: .onoawuaem .xUMEHOUo: .zuceoemenu saw one afloeoe 53m .muoumo nonwe one Hennowao .mouxsooae nonwe one :MHHA:e> .muoumo weave use oum1xo -MHem 1xnuoa .mo>fiuueuuxo Mohave: ensue wee mo>fluoenuxo xuen xwwoso v1“: .nsuzm :hoo .Heuez .Aanu “canneu .mOGO1 .>z .me1e1a 011:2 .ee1 ..ou o1omoz .eeo *oc1 .mwec1 .>z .me1e1e 011;: .mooom Heuocoo .m N.o .womoo mewomeuwe .u sweeuw> .muo>e~u deflowmfiuue one Humane: .oumgmmonm asfiofieouoeoa .owoe ofiama mcwmucou .xwa xcwuv uMOm oocouoozmca common panchu .n «0000 .o oosmm xom .m Acumen xomam Homonx .v QHNHHQGOn—OU 800m HOOK .m mop uemumzm .eoumoz .N e1<-1eox omega .1 mewumwnomoo Humvee; teem .m when .H ucoawhomxm .:o«um:uooow Mono ecu cw com: mausoOHm uoow co newueahowcw weapon; .Hx ofinme 265 ODOR IDENTIFICATION AND INTENSITY Name: INSTRUCTIONS: 1. Try to identify the odor in the beaker by sniffing. 1. SAMPLE: 2. Then, rate the intensity of the odor by putting a line somewhere between no odor and high odor. , IDENTITY: *0 ODOR 2. SAMPLE: F0 ODOR 3. SAMPLE: MEDIUM , IDENTITY: HIE‘} MEDIUM , IDENTITY: HIGH | F0 ODOR 4. SAMPLE: MEDIUM , IDENTITY: HIGH ‘ F0 ODOR S. SAMPLE: MEDIUM , IDENTITY: HIGH—l |NO ODOR Table XII. MEDIUM Scoresheet used in odor recognition and intensity evaluation of Experiment I, Part 3. HIGIT( 266 GENERAL GUIDELINES ON HOW TO RECORD FOOD EATEN IN ONE DAY 1. Milk/fruit juice, and other liquids: Record in terms of measuring cups or ounces. (1 measuring cup = 8 ounces.) 2. Butter: Record as level teaspoon. (1 pat of butter = 1 level teaSpoon. 3. Sugar: Record as level teaspoons. 4. Eggs: Note whether egg or yolk or white only. Cereals and Vegetables Record in terms of measuring cups or tablespoons (1/2 measuring cup equals 8 tableSpoons equals 24 teaspoons). For vegetables If whole, give size (small, medium, and large). If creamed, record in terms of measuring cup or tablespoons, as above. Note whether fresh, frozen, or canned. Meats Record as ounces or level tableSpoons if possible. *1 ounce = 2 level table5poons. Note how cooked. For example, whether it is fried, baked, or broiled. Other foods Record as accurately as possible in terms of ordinary household measures. Note flavor (vanilla, choc. or strawberry, etc.) of a baked product and whether it was iced. Brand names are always helpful. Table XIII. Dietary recording forms used by the college students for their three-day food records in Experiment I. 267 Name: Time: Complete record of the day's food . ------------------------- BREAKFAST------------------------------------ Fruit or juice (circle one) Kind: , Amount: Cereal Kind: , Amount: Milk/with cereal Amount: 02. Sugar: tsp. Bread Kind: , Number of slices: Butter or margarine (circle one used) Amount: tsp. Other Spread Kind: Amount: Egg: Amount: How cooked? Bacon or other meat Kind: Amount: oz. Other foods (please Specify): Kind: . Amounts: BETWEEN BREAKFAST AND NOON MEAL, PLEASE RECORD FOODS EATEN Food (kind) Amount Time Where With Whom COMMENTS: Table XIII, (continued) 268 Name: Time: Complete record of the day's food . ------------------------- NOON MEAL-------------------------------------- Soups, Kind: , Amount: Meat/Main Dish, Kind: , Amount: Potato: How cooked: , Number and size: Vegetable Kind: , Amount: Sandwich filling Kind: Amount: Bread Kind: Amount: Butter/margarine (circle one) Amount on bread: tsp. Salad dressing Kind: , Amount: Milk Kind: , Amount: Other beverages Kind: , Amount: Dessert Kind: , Amount: Other foods, please Specify: Kind: Amounts: BETWEEN NOON AND THE EVENING MEAL, PLEASE SPECIFY WHAT YOU ATE: Food (Kind) Amount ‘Time Where With Whom COMMENTS: Table XIII, (continued) 269 Complete record of the day's food . . . Name: Time: ------------------------ EVENING MEAL---------------------------------- Meat or Main dish: Kind: , Amount: Potato: How cooked? , No. and size: Vegetables: Kind: . Amount: Bread: Kind: , No. of slices: Butter or margarine on bread? Amount: t5p. Gravy: , Amount: Salad dressing: Kind: , Amount: 02. Milk or other beverages: Kind: , Amount: oz. Dessert: Kind: , Amount: OTHER FOODS: Kind: Amount: BETWEEN EVENING MEAL AND BEDTIME, PLEASE RECORD WHAT YOU ATE: Food (Kind) Amount Time Where With Whom Comments: 'Table XIII, (continued) 270 .1 acosvgwaxm :1 muumnasm eo museucw xguumpu cacao; on new: Esau mcpuoo puauu< .>1x e1eeh 1 4.. .1 . i. 1. 1 .. ... . .. 1...... .IIJNI :1 a. _ . ”...-.. _ . . uvucmmd mtammmz Emu” Tummwm . _ . ...... J1... . 18... .18 1.1 Sum... 1.1 31.5... m .35. 88 1.8.15.1... Samoan. iliiiilii.cmnou and atom mcwvou .iiiiiil : 271 Table XV. Actual food items served during the three-day dietary recording of the elderly in Experiment I. Menu for 3 Day Food Intake Analysis* A. Breakfast *Juices (orange, cranberry, prune, apple, grape, grapefruit, pineapple) maple syrup sugar (regular) Sweet 'n Low (substitute) Zesta saltines Keebler-Waldorf low-salt crackers Tea (Salada) coffee (regular) Sanka decaf. coffee CoffeeMate *Milk (whole, skim, Farm Maid cultured buttermilk) hot chocolate (Carnation) butter (regular and low-salt) Fleischman's corn oil margarine fruits (banana, apple, orange, prunes--see lunch/dinner also) Bread (Holsum white and wheat) ketchup/mustard/honey (Nifda brand) cold cereals (Rice Krispies, Cornflakes, Shredded Wheat, Raisin Bran, Special K, All Bran) hot cereals (Nabisco Instant Cream of Wheat, Quaker Instant Oatmeal w/raisins, Malto-Meal) french toast Sausage links eggs (scrambled, poached, sunny side-up, etc. Chef's Pride Frozen brand) danish roll jams (Nifda assorted brands) B. Lunch fruit cocktail soups (Pea, Beef Barley, Cream of Mushroom) **Sloppy Joe (recipe below) *Foods of those who ate out or in their rooms are not included on these lists. **Sloppy Joes 10 1b. Ground Beef (beef and onion) 2 t. dry mustard 8 oz. onion, chopped 2 t. worc. sauce 1 qt. tomato puree 2 t. paprika 2 t. salt 2 t. chili powder 272 Table XV, (continued) cottage Cheese Fruit Plate buttered carrots chocolate pudding Spanish rice w/ground beef tuna salad sandwich tossed salad with bacon bits cherry crisp salad Dressings (French, Gourment--Chadalee Farms, Oil and Vinegar, Creamy Italian, Thousand Island) hot dog and bun w/slice of pickle baked beef hash baked beans spiced peach salad (jello with peaches) cherry cake apple sauce peaches C. Dinner Menu molded vegetable salad (with apricots) swiss steak with gravy liver and onions french fries ice cream (vanilla and chocolate) toppings for ice cream (strawberry and chocolate) wax beans peas (buttered) coleslaw chicken fried steak lamb pattie with mint jelly new boiled potatoes chilled apricots gingerbread with whipped topping cottage cheese beef stroganoff with noodles filet of fish with tartar sauce carrot fingers pears green beans 273 Table XVI. Basic measurements and ladle equivalents used by the Burcham Hills staff to portion-out food items to the elderly during the 3-day dietary food recall. Basic Measurements Scoop #: ngghg: 24 (red) 1-1/2 - 1-3/4 oz. 20 (yellow) 1-3/4 - 2 oz. 16 (blue) A 2 - 2-1/4 oz. 12 (green) 2-1/2 - 3 oz. 10 (ivory) 3 - 4 oz. 8 (grey) 4 - 5 oz. 6 (white) 6 oz. Ladle Equivalents Measure: ngghgg 1/8 c 1 02. 1/4 c 2 02. 1/2 c 4 oz. 1 c 8 oz. 274 Table XVII. Product information on food products used in the odor recognition, Experiment 111 Food Product Description 1. Tuna fish, light chunk 9 1/4 oz. can. Tuna packed in (Sun Harbor Industries) soybean oil, seasoned with vegetable broth and salt. San Diego, Calif. 92113. 2. Tomato paste 12 oz. can. A subsidiary of the (Contadina) Carnation Co., Los Angeles, Calif. 90036. 3. Maple syrup 8 fl. oz. bottle. Grade "A", (Camp) dark amber, 100% natural pure maple syrup. Flessisville, Quebec, Canada. 4. Apple juice 40 F1. oz. bottle. Natural (Mott's) style apple juice, no sugar or preservatives . Duffy-Mott , Co . New York, NY 10017. 5. Garlic powder 2-7/8 oz. canister. Pure garlic (Kroger) powder. Cincinnati, Ohio 45201. 1The three other food items--pepper, lemon, and grape--are described in Study I, and Table XI, Appendix. 275 TO THOSE WHO PARTICIPATED IN THE TASTE SESSION: We would like to take this opportunity to thank you for your time on Tuesday morning. We hope you will join us for additional sessions to begin next Monday, November 5. These sessions will then be held evegy Monday, Wednesday, and Friday between 2:00 - 4:00 p.m. in the Snack E25: Please, we need your help! We need to test you three times next week, once on each of the three days as indicated above. If you can't make it each time, please don't worry. We can always make other arrangements to suit your schedule. We will be calling you soon to remand you about the taste sessions and answer any questions you may have about them or the study itself. Thank you again for your cooperation. This taste experiment means a great deal to us, and_we promise you a "good time". Sincerely, Steve and Mike NOTE: If any of your friends would like to join us next week and were reluctant the first time, please invite them along. It's not too late to get involved! Figure XVIII. Handout distributed to the elderly participants near the end of Experiment 11. 276 From: Steven A. Witherly and Michael D. Dauria Dear Taste Panelist: As you may know, the end of the taste panel draws near, and Mike and I would like to thank you for your participation thus far. All that is left is the salivation testing, which is the most interesting part of the study. It involves the collection of a little hair and saliva. For the saliva study, we will call each of you individually to come to the Snack Bar because we can only handle a few people at a time. The times will be about the same: M-W-F BETWEEN 1:00 and 4:30 STARTING WEDNESDAY, NOVEMBER 28 At the end of the taste study, we hope that you will join us and try the zinc/copper supplement that may improve your senses of taste and smell. All you do is take a tablet of zinc and copper once a day for one month. These tablets are safe, non-toxic, and have no known side effects. Mike and I have taken them for some time now with NO problems whatsoever. We hope that you will continue with the study and try the zinc and copper. Remember you can stop at any time, but we would eatl appreciate your continued support of our project. If you decide to do so, Mike and I will check back with you weekly and see how your taste and smell senses are doing. I appreciate your support of my Ph.D. Dissertation Project. Without you, research and progress involving Senior Citizens comes to an abrupt halt. Thank you again, Steve and Mike Figure XIX. Handout given to all elderly participants of Experiment II, near the end of the olfactory studies. 277 Table XX. Anthropometric data on college students participating in Experiment I and II. Subject Gender1 Age Height Weight Triceps % Fat2 (cm) (kg) skinfold(mm) 1 F 25 168 58 25 33.7 2 F 22 163 52 16 26.7 3 F 22 155 45 18 28.5 4 F 19 170 S7 22 31.6 5 F 23 168 59 21 30.9 6 F 28 163 56 23 32.3 7 F 30 173 61 19 30.6 8 F 34 178 73 24 33.9 9 F 21 160 52 25 33.7 10 F 24 173 $8 72 31.6 11 F 21 157 59 25 33.7 12 F 31 173 59 20 31.4 25.0t4.7 l66.8r7.2 57.4:6.6 lZ.Orl4.8 31.6:2.2 13 M 23 188 82 8 14.7 14 M 25 188 79 9 15.9 15 M 33 180 79 19 23.5 16 M 22 185 79 16 21.8' 17 M 23 183 77 14 20.4 18 M 29 180 61 10 16.9 19 M 25 178 76 12 18.8 20 M 28 178 70 11 17.9 21 M 25 168 54 11 17.9 22 M 24 175 69 20 24.1 23 M 22 185 82 8 14.7 24 M 25 173 6B 17 22.4 25 M 26 188 73 9 15.9 25.4:3.l 180.7:6.3 73:8.4 lZ.6i4.2 18.8i3.3 Mean:DS 25.2:3.9 174:9.8 65.5r10.9 16.616.2 24.917.1 1Female (F); Male (M) 2A5 calculated from equations by Durnim and Womersley (1974), see Appendix, Table XXVI. 278 Table XXIA. Anthropometric data on elderly subjects participating in Experiment I and II. Subject Gender Age Height Weight Triceps - skinfold (cm) (k8) (Iran) 1 F 95 159 40 14 2 F 85 158 48 28 3 F 84 159 41 18 4 F 90 154 61 20 s F 80 158 76 38 6 F 76 156 66 35 7 M1 90 162 68 25 8 F 82 153 68 38 9 F 81 158 75 28 10 F 82 150 44 14 11 F 83 144 58 21 12 F 74 170 57 15 13 F 79 155 69 34 14 F 90 157 80 35 15 F 84 159 83 37 16 F 79 158 66 35 17 F 80 162 66 28 18 F 90 148 49 22 19 F 87 156 65 22 20 F 85 173 74 23 21 F 80 157 53 18 22 F 80 160 50 17 23 M1 80 178 84 14 24 F 79 158 49 17 25 F 82 155 58 25 Mean: 06 83:5.0 158:7.0 62:12.9 25:8.0 1Note: The two males averaged--85. 17.1 years; l70.0rll.3 height; 76.0rll.3 weight; and 19.5t7.8 triceps skinfold. 279 Table XXIB. Anthropometric data on the elderly subjects participating in Experiment III. Subject Age Height Weight Triceps (cm) (kg) skinfold (nun) 25:233. After 3 84 159 41 43.2 18 6 76 156 66 61.2 35 7 90 162 68 70.6 25 .8 82 153 68 69 38 12 74 170 57 57.2 15 15 84 159 83 81.4 37 21 BO 157 53 52.2 18 25 82 155 58 S7 25 Mean:SD 81.5:5.0 158.8i5.3 62i13/6l.5111.9 26.4:9.2 280 fivnmflvxu1muoaoz one awnhnn An oomoHo>oo new xoon mo nofiuoaoonm new nowuenco noflmmouwon no oomem1 1mm-en1 o1ee N 1m~-:e a.mno.m~ m.~11o.~o c.51em1 a.mnc.mm o1e5om me n1eoo1m 1en-m1 . o1ea n1 1m~-=. a.mm «.ono.o1 m.o1nm.mo e.anen1 e.nn~.m~ o1esoe N1 oeo11ou 1uem * ouomnwxm Auxv nae. om< Hoonou macaw mnoownh unwwoz unmmoz .H onenehomnm n“ muoonnnm xauoo1o one munoonum omoafioo mo eueo oflnuonomownune onu mo nomfinemeou .HHxx maneh Table XXIII. 281 medical record survey of the elderly. Are you taking any drugs? No: 6/25 (24%); Yes: 76% Are you taking any vitamins? No: 17/25 (68%); Yes: 32% Are you taking any other pills? No: 25/25 (100%) Do you smoke? No: 25/25 (100%) Do you have any of these illnesses? Diabetes? (2/25: (8%) (l borderline, 1 severe) Kidney Disease? 1/25: (4%) (Kidney stones) High Blood Pressure? 8/25: (32%) (2 cases slight) Answers to the following questions posed in the Other past diseases: arthritis (3/25); skin cancer (1/25) 282 Table XXIV. List of drugs mentioned in the medical record of the elderly in Experiment 1.1 Drug Manufacturer Use(s) Aldactazide Searle Diuretic Antivert Roerig Antihistamine Ascripton Rorer Analgesic Dalmane Roche. Hypnotic Deltasone Upjohn Anti-inflammatory Dilitoxin Purepac Cardiac-Stimulant Dyazide Smith, Kline 8 French Diuretic, Anti- Hypertensive (AH) Hydrodiuril Merck, Sharp 6 Dohme, Diuretic, (AH) Hygroton USV Pharm. Diuretic, (AH) Inderal Auerot Hypertension Lanoxin Burroughs, Wellcome Cardiac-Stimulant Mylicon Stuart Flatulence Notorostat Parke-Davis Myocardial ischemia Peritrate Warner/Chilcott Angina Pectoris Persantine Boehringer Ingelheim Angina Pectoris Prednisone Forest ' Anti-inflammatory Proloid Warner/Chilcott Thyroid extract Quinamm Merrel-National Muscle cramps Stress-Tabs Lederle Vitamins 5 Calcium Thiuretic Parke-Davis Diuretic Triavil Merck, Sharp 6 Dohme Tranquilizer Tranicon Dista 812 intrinsic factor Vasodilon Mead Johnson Cerebral vascular 1Information derived from the Physician's Desk Reference, 1979. 283 Table XXV. Comments by elderly subjects during the course of their Zn/Cu supplementation period in Experiment III. First Week Second Week Subject (Feb. 7, 1980) (Feb. 14, 1980) 1 No change in food taste; pills Maybe more moisture are like "horse pills". in the mouth now. I think it's improved. 2 Might have a change of No change--maybe taste; can't say for sure. more moisture. 3 I haven't noticed anything. Still alive. 4 If we get in the Guiness No problems. Book, I want to be in it. 5 Haven't noticed anything. We're taking our pills. 6 Just taking them regularly. We're taking our pills. 7 No comments. No problems. 8 I haven't noticed anything. Maybe I'm imagining it, but I think I'm smelling better than before. 284 Table XXV, (continued) Third Week Fourth Week Subject (Feb. 21, 1980) (Feb. 28, 1980) 1 Feeling fine, everything For a few days, the tastes the same. 2 Feeling fine, everything tastes the same. 3 Nothing at all. 4 Feeling okay; haven't noticed any special differences. 5 Just taking the pills religiously and don't notice any difference. 6 Just taking the pills religiously, with no changes. 7 Nothing. 8 Can't see any improvement. foods I like taste better. We're just taking the pills. No change. The sense of smell may be better, other than that, just taking the pills. No change--just taking the pills.' No change-just taking the pills. No change. I can't see if there is any difference. 285 Table XXV, (continued) Fifth Week Sixth Week Subject (Mar. 6, 1980) (Mar. 13, 1980) 1 Nothing. No change, but it was worth a try. 2 . Nothing. I don't realize I'm taking anything. 3 Nothing. No--guess not. 4 No. I don't think so. 5 No. Haven't noticed any change at all. 6 I don't know of anything. I have more saliva; have to wipe my lips all the time. 7 No. None that I know of. 8 Can't see if it makes No change; I can't any difference. say there is any difference. 286 Table XXV, (continued) Subject Week Seven (end) March 20, 1980 1 No change. 2 No change. 3 No comment. 4 Not a thing. 5 No change. 6 Still have lgt§_of saliva. 7 No change. 8 No change. 287 Table XXVI. Equations by Durnim and Womersley (1974) used to predict % body fat from a triceps skinfold measurement. Males: 92 S3, 20-29 yr: D = (1.1131 - 0.0530) log TS 34 S3, 30-39 yr: D = (1.0834 - 0.0361) log TS Females: 100 55, 20-29 yr: D = (1.1319 - 0.0776) log TS 58 E, 30-39 yr: 0 = (1.1176 - 0.0686) log TS 48 S3, 40-49 yr: D = (1.1121 - 0.0691) log TS % Fat 3 100 (4.95/0 - 4.50) Example: #1 Female. Triceps = 25 mm Age 25 l) 0 (1.1319 - (0.0776 Log + S (25)) 1.02 % Fat 8 100 (4.95/1.02 - 4.50) = 33.67% Fat Mllilllfilllljllfillflllfllllllfllllllfs