Description and evaluation of a preventive mental health program for preschoolers. A school-based model of preventive intervention is described which identified and significantly overcame school maladaptation among preschool children enrolled in a Detroit Public School Title I Program. Children were screened by teachers for learning and behavioral difficulties using the AML Scale, the Classroom Adjustment Rating Scale, and the Caldwell Preschool Inventory. Sixty-four children were identified on the basis of these instruments as exhibiting some form of school maladaptation. These children were randomly assigned to an experimental condition (N = 32) or placebo control (N = 32). Individually designed prescriptive techniques were developed for high-risk children in the experimental group and were implemented by college student aides after training by the project staff. A multivariate analysis of variance and follow-up univariate analyses, testing differences between experimental and control groups on the posttests, were performed. The difference between the groups on all three criteria measures was significant and favored the experimental children.


The discriminative efficiency of the Bayley Scales of Infant Development. This study was designed to determine the discriminative efficiency of the Bayley Mental and Motor Scales for classifying infants as neurologically suspicious and neurologically normal. The two groups employed in the study were formed on the basis of the results from comprehensive neurological examinations performed at ages 1 and 7. Three discriminant function analyses were conducted, one for each scale separately and a third incorporating both scales using a stepwise procedure. Efficiency was measured in terms of number of correct and incorrect classifications, false positive and false negative errors, and validity coefficients. Comparisons among the analyses were examined by means of relative discrimination power and incremental validity. The findings suggested that the Motor Scale provides the most accurate identification of infants with suspected neurological impairments. A statistically significant likelihood discriminant function derived from both scales yielded some additional discriminative power. The law of parsimony, however, dictated the use of the single Motor Scale.


An adolescent symptom checklist for juvenile deliquents. A rating instrument was developed to assess the personality functioning of problem adolescents who have a brief history of official delinquency. Subjects were 186 residents at a short-term diagnostic detention facility. The instrument that emerged from a factor analysis was tested for reliability, validity, and cross-situational generality. Two major orthogonal factors emerged, Apathy-Withdrawal (I) and Anger-Defiance (II). Each factor was found to be congruent with the corresponding dimension on behavior rating instruments that had previously yielded two orthogonal factors and that had been shown to be relevant to a large number of psychological and educational issues. Significant correlations between the factors and a series of global ratings measuring behavior and measures indicative of the frequency and seriousness of official delinquency provided further evidence of construct and discriminant validity as well as evidence of cross-setting consistency of personality functioning.


An analysis of observational learning in autistic and normal children. The present investigation studied observational learning in autistic children. Fifteen autistic and 15 normal children watched an adult model engage in a set of behaviors under specific verbal instructions. After observing this situation, the children were tested to determine what they had acquired through observation. The results showed that (1) the majority of the autistic and the youngest normal children acquired only some limited features of the observational situation and (2) chronological age was related to the amount of learning through observation in the normal children but not in the autistics. The deficit that the autistic children showed in observational learning may be related to a failure to discriminate or attend to the total stimulus input presented. Their failure in observational learning can be seen to contribute in a major way to the severely impoverished behavioral repertoires of these children.


Test anxiety and evaluation threat: children's behavior in the classroom. Children differing in test anxiety level were observed in two art classes, one immediately preceding a classroom examination, the other when no examination was expected. The observational analyses indicated the following: (a) When an examination was expected there were general increases both in children's task orientation and in their concern with other's evaluations, and a decrease in general activity level. It was suggested that future research examine the effects of additional situational variables on children's classroom behaviors. (b) Highly test-anxious children were generally hardworking and inactive. They reacted to examination expectancy with a decrease in task orientation, reduction in communications, and attentional constriction. Less anxious children reacted to examination expectancy by working harder, eliminating task-irrelevant behaviors, and seeking feedback. These results were interpreted as supporting a cognitive theory of test anxiety.


Behavior problems of preschool children: dimensions and congenital correlates. Factor analysis of a revised Behavior Problem Checklist for a sample of preschool children yielded six oblique primary factors and two orthogonal second-order factors. Scores on these factors were correlated with activity level, gross- and fine-motor incoordination, minor physical anomalies, and sociability. There were sex differences and factor differences in the resulting patterns of correlations. The patterns of correlations also indicated that the six primary factors could be collapsed into two broad (Conduct and Personality) and two narrow (Distractibility and Attention Seeking) factors.


Relationship among environmental and demographic variables and teacher-rated hyperactivity. This study assessed the role of environmental and demographic factors in the occurrence of teacher-rated hyperactivity. The subject group consisted of 79 hyperactive and 81 nonhyperactive children ranging from 5 to 12 years of age. Parents of the subjects were interviewed to obtain information regarding the environmental and demographic factors of sex, race, birth order, number of siblings, frequency of change of residence, income level, mother's age, father's age, educational level of mother, educational level of father, parents' marital status, and the method of child discipline used in the home. Comparison between the the hyperactive and nonhyperactive groups suggested nonsignificant differences with the exception of sex (p less than .001), in which the ratio of hyperactive males to hyperactive females was 5:1.


An interview with a child to assess psychiatric disturbance: a note on its reliability and validity. Forty-six children with enuresis were given a psychiatirc interview. The two writers made independent ratings of 10 behavioral variables. Seven of these showed satisfactory interrater reliability. No relationship was established between child psychiatric disturbance assessed in this way and estimates of disorder obtained from information given by mothers and teachers.


A psychosocial study of hyperactive and learning-disabled boys. Adequately reading hyperactive boys, normally behaved learning-disabled (LD) boys, and normal controls were contrasted on tests measuring personality traits, cognitive role taking, and moral resoning. Additionally, parents and teachers rated all children on a number of behaviors, and parents were interviewed in a process-oriented fashion to assess home stimulation potential. Hyperactive boys were rated more aggressive and anxious than LD boys and controls and had not been encouraged as much by parents to achieve. Hyperactives had been born to younger parents, on the average, and 25% lived with their mothers and stepfathers. None of the LD or control boys had stepfathers. The groups did not differ significantly in moral reasoning ability, cognitive role taking, or locus of control; on the Junior Personality Inventory hyperactives tended to have elevated scores on the neuroticism scale while LD boys had higher scores on the lie scale.


Food and nutrition policy: probability and practicality. The nutrition community has, unfortunately, tended to overemphasize the benefits to be derived from consumption of recommended levels of essential nutrients and has tended to ignore or underemphasize--except for obesity--the effects of over-consumption. New guidelines incorporating both concepts must and will be formulated. The nutrition community has the opportunity to provide the leadership required. If it does not meet the challenge, that will be assumed by others.


The scientific basis of the "Goals": what can be done now? Universal dietary goals for the general public cannot be formulated or implented. More appropriate would be guidelines to serve as preventive measures for specific groups, based on genetic endowment, age, sex, and condition. Nutrition education as a component in health education is essential.


Are professionals jumping the gun in the fight against chronic diseases? The first steps in implementing a national nutrition policy of providing a food supply which meets the nutritional requirements of our population have been the periodic issuance of the Recommended Dietary Allowances. More recently, the "Dietary Goals for the United States" have been released by the Select Committee on Nutrition and Human Needs of the U.S. Senate, developed in the belief that they would, if followed, serve as one avenue of preventing coronary heart disease. The author presents a "critique" of the Dietary Goals, including a re-examination of food consumption statistics and a summary of intervention trials in five countries in which diet or drug therapy was aimed at lowering serum cholesterol. Listed are four criteria for evaluating recommendations to the public about diet and chronic disease. Rather than making a major revision in the proportion of fat and carbohydrates in our national diet, he recommends only limitation of the overall amounts to achieve weight control for those who need it.


Are low-cholesterol diets expensive?

Are low-cholesterol diets expensive? To serve as the basis of cost comparison, USDA "moderate-cost" and "thrifty" menus for one week were modified to meet guidelines for a cholesterol-lowering diet. Prices of individual items on the menus at both cost levels were obtained from two supermarkets and averaged. Items on the moderate-cost, cholesterol-lowering diet were $1.37 less than the USDA menus. At the low-cost level, the modified menus cost 22 cents more. In neither case was the difference significant by the t-test.


Cholesterol Index of foods. Teaching tool for fat-modified diets. The Cholesterol Index of foods expresses the relative effect of individual portions of foods on the serum cholesterol of an average individual. It is calculated from food cholesterol content and fatty acid composition. The total Cholesterol Index for the day has a value of 100 for a 2,400-kcal, prudent diet and a value of slightly above 200 for an equicaloric average American diet. The Cholesterol Index for a recipe prepared at home is the sum of the Index values for its ingredients. The Index can be adapted to individual dietary prescriptions fo patients with hyperlipidemia. After publication of Cholesterol Index tables, the Index could be used by dietitians or by individuals who wish to alter their present food pattern to a more prudent diet.


Environmental fate of rice paddy pesticides in a model ecosystem. The distribution and metabolic fate of several rice paddy pesticides were evaluated in a modified model ecosystem. Among the three BHC isomers, beta-isomer was the most stable and bioconcentrated in all of the organisms. Alpha- and gamma-isomers were moderately persistent and degraded to some extent during the 33 day period. Disulfoton was relatively persistent due to the transformation to its oxidation products. Pyridaphenthion was fairly biodegradable. N-Phenyl maleic hydrazide derived from the hydrolysis of pyridaphenthion was not detected in the organisms though it was found in the aquarium water after 33 days. Cartap and edifenphos were considerably biodegradable, and the ratio of the conversion to water soluble metabolites was very high. There was a distinct difference in the persistence of Kitazin P and edifenphos in the aquarium water. It appeared that the hydrolysis rate of the pesticides affected their fate in the organisms. PCP appeared to be moderately biodegradable. CNP was considerably stable and stored in the organisms though the concentration in the aquarium water was relatively low. The persistence and distribution of the pesticides in the model ecosystem were dependent on their chemical structures. In spite of the limitation derived from short experimental period, the model ecosystem may be applicable for predicting the environmental fate of pesticides.


Effects of dieldrin on operant behavior of bobwhites. Bobwhites (Colinus virginianus) were trained to peck a green lighted key to receive food. Bird response performance was monitored for 14 days, then birds were dosed with five levels of dieldrin (50 to 300 micrograms every other day) for 42 days while their response performance was monitored. At the end of the 42-day dosage period, mean brain concentrations of dieldrin ranged from 2.6 ppm to 11.8 ppm; lower concentrations associated with lower dosage levels. All dieldrin dosage levels altered cage behavior of bobwhites, dieldrin dosage levels greater than 100 micrograms resulted in slower and less accurate responses by dosed birds during the 42-day trial period.


Photomirex: a teratogenicity and tissue distribution study in the rabbit. Adult New Zealand white does were intubated orally with single daily doses of 0, 5, or 10 mg of photomirex (8-monohydromirex) per kg body weight from the 6th through to the 18th day of gestation. Pregnancies were interrupted at term by cesarian section and fetuses removed and evaluated by following routine teratologic methods. Both maternal and fetal tissues were analyzed for residues of photomirex. None of the treated does showed any sign of toxicity. Except for a significant reduction in the mean fetal weight of the 10 mg/kg group all other parameters which evaluated fetal survival and fetal development were within the control range. Photomirex was found in all tissues examined. In the doe, the highest levels were found in fat followed by liver, kidney, spleen, heart, brain and blood. Photomirex was readily transferred across the placenta and accumulated in the fetus. However, in the fetus the highest levels were found in the heart, followed by liver, brain and blood. There were no teratogenic effects at the doses used in this study.


Persistence and movement of chlorbromuron in potato soil. Movement and persistence of chlorbromuron applied at rates of 1.5, 3, and 6 kg a.i./ha was studied in a New Brunswick potato soil for one growing season. Most of the chlorbromuron remained in the 0 - 2.5 cm soil depth with slight residues in the 2.5 - 5, 5 - 7.5, and 7.5 - 10 cm depths. After 47 days the chlorbromuron residues had dropped to 40%, then gradually levelled off to 25% at the end of the season. Chlorbromuron was determined directly by electron capture gas chromatography using a short glass column of 3% OV-210.


Determination of disulfoton and permethrin residues in an organic soil and their translocation into lettuce, onion and carrot. The residues of disulfoton and permethrin in an organic soil and in vegetables grown in soil treated with a granular formulation of the pesticides were determined by gas chromatography. The residues were removed from soil or plant samples by successive extractions with acetone and hexane. Permethrin persisted in the soil for the initial 28 days and declined slowly during the rest of the season but disulfoton after persisting for one week at the applied concentration was degraded in the next two weeks. The insecticides did not translocate into the edible parts of the vegetables but were present in the root system of onion and lettuce. Carrot and lettuce yields were not singificantly different from those of the controls but onion yields were substantially decreased by the use of permethrin.


Effects of sub-chronic low-level dietary intake of chlordane on rats with cirrhosis of the liver. Male rats, 60 days old, were treated with chlordane during or after induction of liver cirrhosis with carbon tetrachloride to determine the effect of treatment with chlordane on the response of the rats to the disease. When liver cirrhosis was induced simultaneously with chlordane treatment the disease symptoms were aggravated; the lipid content of the tissue was lowered significantly, growth rate was significantly lower than controls and there was no apparent replacement of damaged liver tissue by liver growth. The cytochrome P450 content of the liver was similar after both treatments. Continuation of the chlordane treatment after termination of the carbon tetrachloride treatment brought about a more rapid recovery from the induced cirrhotic condition. All these responses were to a dose range one tenth the recommended "no effect" level for healthy animals.


Delayed neuropathy in sheep by the phosphonothioate insecticide cyanofenphos. Cyanofenphos (surecide)(R), 25% E.C., O-ethyl O-(4-cyanophenyl) phenylphosphonothioate, was orally administered to one year old lambs at sublethal doses of 1 mg, 2 mg and 4 mg active ingredient kg-1 day-1 for time intervals 60, 45 and 30 days respectively. Irreversible paralytic ataxia symptoms of delayed neuropathy appeared at about 80, 50 and 30 days respectively. In weekly blood samples, AChE (acetylcholine-sterase) and MAO (monoamine oxidase) activities were inhibited depending upon level of dosing and time interval. However no significant correlation was found between the extent of plasma AChE and MAO inhibition and the onset of ataxia symptoms. In brain samples from ataxiated animals, AChE, MAO and NTE (neurotoxic esterase) activities were assayed simultaneously with untreated animal. Direct correlation was shown between in vivo NTE inhibition and the occurrence of delayed neuropathy. Cyanofenphos is the third compound of the phenyl phosphonothioate type on the market showing delayed neuropathy together with Leptophos and EPN.


The metabolic fate of N-isopropyl-N-phenyloxamic acid in the rat and the milk goat. Rats excreted the 14C from a single oral dose of N-isopropyl-N-[14C]phenyloxamic acid [I, a soil metabolite from 2-chloro-N-isopropylacetanilide (propachlor)] in approximately equal quantities in the urine (49.2%) and feces (48.2%). A milking goat given daily oral doses of [14C]-I (1 mg of I three times daily) excreted more 14C in the feces (56.6%) than it excreted in the urine. From both species, I accounted for 97 to 100% of the urinary 14C, and all of the 14C that was extractable from the feces (73 to 75% of the 14C in feces was extractable with methanol). Goat milk samples collected 16 hr after the last dose contained no detectable 14C. Tissue residues of 14C were determined.


Fate of ochratoxin A in goats.

Fate of ochratoxin A in goats. The fate of ochratoxin A (OA) was studied in goats given a single oral dose of 3H-OA (0.5 mg/kg). More than 90% of the radioactivity was found to be excreted in 7 days and the majority (53%) was found in feces. Thirty-eight percent, 6% and 2.26% of the activity was found in urine, milk and serum, respectively. The radioactivity in the liver and kidney 6 hours after feeding amounted to 1.5 and 0.5% of the total dose administered, respectively. Subsequent fractionation of liver and kidney homogenates revealed that microsomes, ribosomes and post-ribosomal supernatant fractions contained most radioactivity. Thin layer chromatographic analyses revealed two additional radioactive spots with Rf values and fluorescent characteristics different from OA, Oalpha and 4-OH-OA. Whereas OA was found as the unaltered molecule in feces, the metabolites were primarily found in urine and milk. Less than 0.03% of free OA was found in milk during the 7-day period.


Effect of ferrous sulfate on parathion degradation in flooded soil. In an isotope study, the effect of ferrous sulfate on the degradation of parathion was studied under flooded soil conditions. The addition of ferrous sulfate to flooded soil led to more rapid and extensive degradation of parathion with the formation of additional degradation products in ferrous sulfate-amended soil. This effect was not pronounced when ferrous sulfate was added to non-flooded soil or to flooded autoclaved soil. Sulfate, rather than Fe2+, was implicated in the extensive degradation of parathion.


Blood-brain barrier permeability in senescent rats. Cerebrovascular permeability to 14C-sucrose was measured in 3-mo-old and 28-mo-old conscious, restrained Fischer 344 male rats. Tracer was injected intravenously in these animals and arterial plasma concentrations were followed for 50 to 240 min, when animals were killed and regional brain radioactivity was measured. Equations derived by a two-compartment diffusion model were fit to concentration data to estimate PA (product of cerebral capillary permeability and surface area) and Ve (cerebral distribution space of 14C-sucrose). PA in 3-mo-old rats averaged 7.53 x 10(-6) sec-1 in 14 cerebral regions, and was not significantly elevated in 28-mo-old rats except possibly at white matter. Ve fell from an average of 0.126 in control rats to 0.070 in 28-mo-old rats. This 45% reduction may reflect a reduced extracellular space in the older animals. The findings do not support the hypothesis that aging of the central nervous system is related to breakdown of the blood-brain barrier.


Trypsinization frequency and loss of proliferative capacity in WI-38 cells. The effect of the trypsinization procedure employed in routine serial subcultivation on the proliferative life span of WI-38 cells was studied by varying the frequency of trypsinization and by using a nonenzymatic subcultivation technique. The results indicate that recurrent trypsinization does not contribute to loss of proliferative capacity and may, in fact, delay this loss to some degree.


Sequential appearance of auto-erythrocyte antibody immunoglobulin classes with age. The autoimmune, short-lived, NZB/BINJ mice were followed for the changes taking place in the number of thymic and splenic nucleated cells from day 1 up to 12 mo of age. The long-lived C57BL/6J mice were monitored in parallel with the NZB mice to see whether similar changes were reflected. Both strains of mice were investigated for the sequential appearance of anti-erythrocyte autoantibody classes by the direct Coombs' agglutination test. IgG1 was the first class of antibody detected on the erythrocytes and was followed by IgG3, IgG2 and IgA, simultaneously. IgM was the last antibody to react with erythrocytes. The C57BL strain had a very small number of mice with low levels of IgG1 antibody on erythrocytes throughout the study.


Heart rate and skin conductance during cognitive processes as a function of age. Heart rate (HR) and skin conductance (SC) were measured in males ranging in age from 23 to 87 years while they were performing mental arithmetic tasks, subtractions, and during multiple trial free recall. Performance on the arithmetic tasks was not affected by age. HR but not SC showed greater changes from basal levels in young subjects. Free recall was greater in young than in old subjects, and changes in HR were again greater in the young subjects. During intertrial rest periods HR returned t9 basal levels in young but not in old subjects. These changes are similar to those seen in older people after physical exercise. Correlations between HR and SC changes during arithmetic performance and during free recall indicated that response stereotypy is to some extent age dependent.


Anxiety, aging and the third emergency reaction. Severe anxiety, unlike severe depression, is not common among the aged. In view of the multitude of stresses accompanying later life and considering the fact that lack of success in mastering stress typically produces anxiety, one should expect a high frequency of anxiety among the aged. To explain the deficit, it is proposed that those who survive into old age have developed strategies to successfully deal with stress, that among these strategies is a passive stance, termed "freeze," and that "freeze" is a third emergency reaction, one which was omitted from the fight-flight paradigm of emergency reactions.


Participation of senior citizens in the swine flu inoculation program: an analysis of Health Belief Model variables in preventive health behavior. Evidence is presented of statistically significant and strong relationships between Swine Flu Inoculation status and nine variables in the reformulated Health Belief Model with 122 randomly selected subjects, primarily Black and Portuguese-American, who are active members of two Providence, Rhode Island senior centers. No statistically significant relationship was discovered between inoculation status and previously having had the flu. The variables which were found related are: Efficacy, Safety, Knowledge of Side Effects, Prior Flu Shot Status, Proportion of Friends and Relatives Who Got the Shot, Sex, Race, Future Plans for Flu Shots, and Future Plans for Other Inoculations. The data were obtained through personal interviews in the Spring of 1977. It is suggested that the results provide some basis for optimism for successful intervention designed to change the future preventive health behaviors of nonparticipants in the Swine Flu Inoculation Program. Many nonparticipants had fears and doubts about the effectiveness and safety of the shot and are amendable to suggestions from physicians about future inoculation participation. Full information should be provided to high-risk groups such as senior citizens about the relative risks of suffering serious side effects, the effectiveness and safety of the procedure for persons their age with the typical health problems of senior citizens, and the relative risks and dangers to them of contracting the illness against which the shot is designed to protect them.


The influence of stimulus age and sex on person perception. College males and females evaluated 25-, 50-, and 75-year-old males and females using a semantic differential. Evaluations became more negative with increasing age of stimulus concept. Males were perceived as more effective and autonomous than females until age 75. Females were perceived as more personally acceptable at all ages. The aging process was perceived as similar for males and females between 25 and 50 but more detrimental to males than to females between 50 and 75. Female Ss evaluated male and female stimulus concepts similarly while male Ss evaluated male stimuli more positively than female stimuli.


Differences in the magnitude of the Delboeuf Illusion and Usnadze Effect during adulthood. The Delboeuf illusion (Type I) and Usnadze Effect (Type II) were used to investigate changes in perceptual processes in 72 female subjects, aged 20-79 years. Unlike other studies, small figures (less than 2 degrees) were presented for brief periods under fixation conditions. The magnitude of the Delboeuf illusion increased in old age, while the magnitude of the Usnadze Effect showed no change.


Age-related differences in two stages of central perceptual processes: the effects of short duration targets and criterion differences. Two experiments investigated age differences in speed of central perceptual processes using a dichoptic backward masking paradigm. Twelve young (mean age = 19.6 years) and 12 old (mean age = 68.5 years) adults served as subjects. Exp. 1 examined age differences in speed of central perceptual processes using an ascending limits procedure and forced-choice responding to control for criterion differences. Target durations from 2 to 30 msec were used to further test the fit of young and elderly masking functions to the additive rule believed to describe masking which arises centrally (Turvey, 1973). Results from Exp. 1 showed a 33% slower speed of processing in the elderly sample. Furthermore, masking functions for both age groups fit the central rule across the wide range of target durations used. Exp. 2 examined the perceptual detectability of a 2- and 20-msec target at 7 stimulus onset asynchronies between target and masking stimuli using a forced-choice procedure. Subanalysis of two different components of the observed masking functions suggested the elderly sample was equally slow in completing both an early and late stage of central perceptual processing.


A long-term longitudinal study of intellectual ability of older adults: the matter of selective subject attrition. Two hundred and forty-six subjects aged 60 to 94 were tested with the WAIS at test session 1, the start of a longitudinal study which lasted approximately 20 years. The subject attrition from the first session to the last (session No. 11) was progressive and selective, leaving mainly the intellectually superior ones in the study. Two facts appeared particularly important: One, little intellectual decline was seen in these superior subjects until very late in life. Two, analyses based on such superior subjects without reference to the larger body of subject populations can lead to spurious generalizations about aging in general. Age decline in intellectual ability seen in more representative populations may not be apparent when analyzing data of select samples.


WAIS performance for different age groups of hypertensive and control subjects during the administration of a diuretic. This study examined the effects of age and hypertension on WAIS performance. Previous studies have reported that hypertension adversely affects cognitive functioning, but few have considered the influence of hypertension on discrete age groups. Hypertension was found to be negatively associated with WAIS Verbal scores for younger (21 to 39 years) but not older (45 to 65 years) subjects. Further, younger subjects attained higher WAIS Performance scores than did older subjects. Medication history was not associated with performance levels for hypertensives, nor was performance of controls influenced by whether or not they were tested while on a diuretic. When WAIS Performance scores were analyzed for subjects matched on WAIS Verbal scores across age (21 to 39 vs 45 to 65 years) and blood pressure (normotensive vs hypertensive), a significant age by blood pressure interaction was found. The effect of blood pressure on Performance scores was found to be greater for younger than for older subjects. The association between plasma renin activity (PRA) level and WAIS Performance scores was also examined. Contrary to previous findings PRA was not related to Performance scores. The results are discussed in the context of previous hypertension research and their implications for comparisons across age levels.


Pre-retirement life-style and the degree of planning for retirement. The purpose of this study was to examine the relationship between the pre-retirement life-style of adult men, and both the degree of planning for the retirement years and the decision to retire early. Adult males (N = 360) between 55 and 64 years of age were interviewed about their present leisure life-style and the amount of retirement planning they were doing or had done. In order to determine which demographic, attitudinal and social participation variables predict retirement related behavior, Multiple Classification Analysis (MCA) was used. It was found that socioeconomic status, health, involvement in expressive type organizations, job satisfaction and degree of leisure orientation were associated with pre-retirement attitudes toward retirement.


Age and the satisfactions from work.

Age and the satisfactions from work. This research documents age differences in the relative importance of intrinsic work satisfactions as determinants of the individual's sense of global well-being. Evidence of a decline in the importance of these intrinsic satisfactions for men during the later stages of labor force participation is not an artifact of lower levels of satisfaction. Rather, it appears that toward the end of the period of labor force participation, the satisfactions men derive from work are transferred from the actual experience of work to its consequences. The relevance of the results to early retirement policies is considered. Data used in the analysis come from the 1971 Quality of American Life survey. The national cross-sectional survey allows for comparison of the multivariate structure of life satisfactions among four age groups of labor force participants (i.e., 21-34, 35-44, 45-54, 55-64).


Differentials in survivorship of the older populations among areas within a state. The case of Pennsylvania. It has often been assumed when making estimates or projections of populations of sub-state areas that survival ratios of the state, region, or even the nation may be used since significant differentials are not expected within the state. The validity of this assumption is questioned, employing single-year-of-age survival ratios estimated for each planning and service area in Pennsylvania. Regression analysis estimations were made to determine whether the survival functions of the areas were statistically the same as the survival functions of the state. The survival functions were partitioned into two groups by age; a 10 to 49 year age group and a 50 to 84+ year age group. Briefly, the results indicated that, at least for Pennsylvania significant differences existed and that these differences are more likely to occur among the older group.


Recovery of sensibility after suture of digital nerves. A group of 49 patients with 74 injured digital nerves was studied more than 5 years following epineurial nerve suture. Three tests of return of sensibility were used: von Frey plastic filaments, a modified Weber two-point discrimination device, and a dynamic test--the plastic ridge. The correlation between two-point discrimination and tactilegnosis in the range of 8 to 12 mm was poor. The plastic ridge device detects the presence or absence of tactilegnosis in patients in the intermediate range of two-point discrimination between 8 and 12 mm. Of the factors studied, the patient's age at the time of nerve suture was found to be directly related to the return of sensibility by both two-point discrimination and ridge criteria.


Skeletal deformities of the denervated hand in Hansen's disease. Skeletal deformities of the denervated hand are subtle in their development and pose a more formidable problem than do primary deformities of Hansen's disease. Basic osseous changes consist of concentric, longitudinal, or the longitudinal varying methods of absorption. Knowledge of their pathogenesis provides the basis for their management. Prevention of deformity and conservative management of secondary complications are necessary to salvage as much of the hand as possible.


Ray transposition for central digital loss. In fourteen cases transposition of either the second or fifth ray was used following traumatic amputation of one or both central digits, or in conjunction with surgical removal of either the middle or ring finger when it was rendered useless by trauma, involved with a malignancy, or congenitally hypoplastic. The operative technique consisted of osteotomies at the base of the metacarpals with internal fixation of the transposed ray. When the fifth ray was transposed, the level of division of the bone was such as to provide the appearance of a more normal length of the digit.


Compartment syndromes of the forearm: early recognition using tissue pressure measurements. A compartment syndrome of the forearm can be a devastating injury if not relieved promptly by early fasciotomy. Of five patients who developed compartment syndromes, compartment pressures were measured in four and found to average 69 mm Hg. Tissue pressure measurements provided early objective evidence of the presence of the compartment syndrome and contributed significantly to the ultimate functional recovery. We would advise repeating the measurement for pressures in excess of 30 mm Hg in the upper extremity, and fasciotomy for pressures in excess of 40 mm Hg when accompanied by any neurovascular compromise.


Fibromatosis of the upper limb.

Fibromatosis of the upper limb. Two cases of fibromatosis of the upper limb are presented, neither of which could be placed in a specific category as described in the literature. Both patients were black, and both had a history of previous trauma to the region, which has been regarded as an initiating factor. Emphasis is placed on conservative treatment in order to maintain hand function as long as possible.


Iterative image transformations for an automatic screening of cervical smears. The new generation of image analysis systems permits the use of iterative image transformations. It is now possible to construct algorithms where the elementary steps are not arithmetic operations but image transformations. This will be illustrated by two examples. In the first, the absorption image of Feulgen Stained nuclei is processed by contrast algorithms in order to detect suspect cells. In the second, free lying cells are separated from overlapping cells and other artefacts by the use of skeletonization procedures.


Transformation of nuclear morphology during cellular maturation. Using in vivo labeling of mouse leukocytes with tritiated thymidine, cells in the neutrophilic series were studied to determine the change in their nuclear shape as a function of maturation level. Several morphologic shape parameters including perimeter and bending energy were used to quantify the distribution of the nuclear morphology in a given age cohort. The change in these distributions as a function of calender age level was determined. The two parameters named above were used to test the possibility of inferring the age from the quantitative morphology.


Light scattering and fluorescence by small particles having internal structure. We consider two related, yet distinct queries: 1. How does the internal morphology of a small particle affect the elastic light scattering signals? We have devised an algorithm, presently accurate for particles comparable only to small biological spheres (diameter less than 1 micron), which suggests that light scattering is sensitive to internal morphology only in the backward directions. Accordingly, observations should be obtained in these directions when probing for internal morphology. 2. How are fluorescent signals affected when the active molecules are variously distributed within small particles? One cannot assume that the fluorescent signals are simply proportional to the number of active molecules contained in the particle because there may also be a dependence upon the geometrical and optical properties of the particle and upon the particular spatial distribution of these molecules within the particle. Indeed, even the measured emission spectrum may be affected by such morphological features. Here, too, these calculations are mainly restricted to small particles (diameter less than 1 micron) in which the fluorescent molecules are isotropic and immobile. Under these conditions the effects are quite dramatic. These effects should be considered in quantitative procedures which utilize fluorescence for determining the concentration of specific molecules in small particles such as biological cells. They may provide a clue for discriminating among cells which differ morphologically or in which the spatial distribution of the fluorescent moiety differs. These effects may be minimized by utilizing a light source which is polarized perpendicularly to the scattering plane.


Cell volume decrease during Friend leukemia cell differentiation. Friend leukemia cells (GM86, clone 745) were induced to differentiate with dimethyl sulfoxide or butyric acid. The kinetics of induction were measured by cell growth, cell volume distributions, and [3H]thymidine incorporation. From the volume distributions, it was found that the rate of induction was both agent sensitive and concentration dependent. The changes in volume distributions occurred approximately 4 hr earlier with dimethyl sulfoxide induction relative to butyric acid induction. However, the changes with the butyric acid induction were more dependent on concentration. A decrease in labeling indices during the 12- to 20-hr time period was correlated to a decrease in mean cell volume and an increase in the proportion of G1 cells. After the 20-hr time period of induction, an increase in labeling indices and in the percentage of large cells was observed. The data suggest that a transient block of cells in G1 occurred between 12 to 20 hr, and that the early differentiation involved a volume decrease which was related to a redistribution of cell cycle stages. The study was also shown that the changes of cell volume are a rapid monitor to determine the early events of the differentiation process.


Cellular autofluorescence--is it due to flavins? For many cell types, cellular autofluorescence in the 500-600 nm spectral region can result in a significant background signal for measurements of weakly fluorescent probes. Measurements of fluorescence intensity and spectra of a variety of cell types and assays of neuronal homogenates are presented to demonstrate that this autofluorescence is most likely due to endogenous flavoproteins.


Polarization of fluorescein fluorescence in single cells. Measurement of fluorescence polarization (P) gives information about the immediate environment of the fluorescent molecule. We used a flow polarimeter to investigate the factors influencing P of fluorescein in mammalian cells to determine whether such measurements are useful for characterizing heterogeneous cell populations. Fluorescein was introduced into cells by incubation with FDA. Measurements of the intensity of fluorescence (TI) and polarization (P) revealed an unexpected dependence: P decreased with increasing intensity of fluorescence. This may be accounted for by the classical model of the binding of small molecules to protein in which P is dependent on the ratio bound to unbound molecules. We have been able to estimate the quenching due to binding and construct a Scatchard plot. We estimated a wavelength shift from in vitro data consistent with the dependence of P on wavelength seen in our cell work. Generally, the distributions of P are symmetrical. Photon statistics broadens the P distribution of dim cells. However, structure does develop in the P distribution when the cells are deprived of calcium or incubated in the cold. This appears as a shoulder on the P distribution or resolves into two peaks. Calcium deprivation may differentially affect a subpopulation of cells whose significance remains to be explored in various cell types.


Centrifugal separation of cells in sputum. A centrifugation technique was developed and used to separate fixed cells from the sputa of patients with bronchogenic squamous cell carcinoma and ones with no evidence of cancer. This article presents the relative frequencies of occurrence of five cell types (i.e., leukocytes, macrophages, squamous, columnar and atypical/cancer) in specimen fractions separated from a discontinuous aqueous Ficoll density gradient. These differential counts show that individual cell types may be selectively collected. Atypical and cancer cells are found at high-density gradient fractions (p congruent to 1.138-1.155 g/ml) with a 10-fold enrichment over unprocessed samples.


A microplate immunoenzyme assay for anti-influenza antibodies. An enzyme-linked immunosorbent assay (ELISA) using horseradish peroxidase is described for the detection and quantitation of anti-influenza virus antibodies. Compared with complement fixation and hemagglutination inhibition tests, ELISA is far superior with respect to sensitivity and reliability. Non-specific viral inhibitors present in sera do not affect the titer in ELISA. Its sensitivity, close to that of radioimmunoassay permits detection of small amounts of antibodies in pulmonary secretions and supernatants from in vitro spleen cell cultures.


Methods for binding cells to plastic: application to a solid-phase radioimmunoassay for cell-surface antigens. Two methods are described for attaching cells to plastic plates such that they may be used for antibody binding assays. In the first method, lymphoid cells or erythrocytes were attached to the wells of plastic plates using glutaraldehyde. This resulted in monolayers of fixed cells which retained surface antigens and were stable to storage. The second method involved binding of unfixed cells to the plastic surface by means of antibodies non-specifically adsorbed to the plate. Both methods resulted in cell layers which remained attached to the plate during the washing and incubation procedures of a radioimmunoassay. The cell layers were shown to be suitable for screening the product of hybrid cell lines for the presence of monoclonal antibodies to cell-surface antigens.


The effect of cryopreservation of lymphocytes on E rosetting ability: a study in lung cancer patients and controls. Two methods of measuring total T cell percentages have been used to assess the effect of freezing and storage on the ability of lymphocytes to form rosettes in blood transfusion donors, laboratory staff, patients awaiting surgery for non-malignant conditions and 48 preoperative lung cancer patients. A significant reduction in rosetting ability after freezing was found for the surgical patients group but not for the other groups. With frozen lymphocytes we have found the same significant decrease in the percentage of T cells seen with fresh lymphocytes when lung cancer patients are compared with the blood donor group. A significant difference in the percentage of T cells detected by the two assays with frozen lymphocytes was found in the control groups.


Recovery of soluble feline oncornavirus-associated cell membrane antigen from large volumes of tissue culture fluids. Feline oncornavirus-associated cell membrane antigen (FOCMA), which is released spontaneously in tissue culture, was harvested from large volumes of culture fluid. Continuous-flow molecular filtration was the method of choice for concentrating the component. The yield of antigen recovered, as determined by cytotoxicity inhibition, was equal to that recovered with concentration in cellulose membrane tubing against polyethylene glycol and half that obtained by lyophilization. The latter two techniques were found to be impractical for handling large volumes of material. However, lyophilization was used for the final concentration step following volume reduction to approximately 300 ml in the molecular filtration system. Antigen yields were equivalent when removing salt by constant-volume molecular washing, prior to concentration in the filtration system, or when removing salt by conventional diffusion through cellulose membrane tubing.


Circulating immune complexes in experimental streptococcal endocarditis: a monitor of therapeutic efficacy. An important problem in the management of infective endocarditis has been the delineation of laboratory procedures that are sensitive, reliable indicators of therapeutic efficacy. Because circulating, complement-containing immune complexes of the IgG type (CICs) have been demonstrated in most humans with infective endocarditis, serum CIC levels during the natural course of the infection and in response to penicillin therapy were studied in 42 rabbits with right-sided endocarditis due to Streptococcus salivarius. A significant rise in the level of CICs in both 21 control rabbits and 21 treated rabbits was observed after induction but before treatment of infective endocarditis (P less than 0.01). In the 17 successfully treated rabbits, CIC levels fell sharply during the first week of therapy and remained at preinduction levels thereafter (P less than 0.005). In contrast, CIC values did not change significantly either in control animals or in the four treated animals with refractory endocarditis, although in the latter animals, serum bactericidal titers remained less than or equal to 1:32. These findings suggest that serial measurements of CIC levels during antimicrobial therapy of infective endocarditis may aid in monitoring therapeutic efficacy.


Plasmid-mediated, transferable resistance to clindamycin and erythromycin in Bacteroides fragilis. A clindamycin- and erythromycin-resistant strain of Bacteroides fragilis, TMP10, that had been isolated from a blood culture transferred its drug resistance in a Millipore mating procedure to another strain of B. fragilis, TM2000, and to a strain of Bacteroides thetaiotaomicron. Donor strain TMP10 contained numerous species of extrachromosomal DNA ranging from approximately 2 x 10(6) to 20 x 10(6) daltons in molecular weight, as revealed by agarose gel electrophoresis. Clindamycin- and erythromycin-resistant transcipient cells of B. fragilis strain TM2000 always contained at least two plasmid species with molecular weights of 2 x 10(6) and 20 x 10(6) daltons, respectively, although other plasmid species were also transferred.


Enterobacter cloacae septicemia in a burn center: epidemiology and control of an outbreak. An outbreak of infections due to Enterobacter cloacae occurred in the burn center at the Medical College of Virginia (Richmond, Virginia) in 1976. Fifteen patients had bacteremia due to E. cloacae; 10 cases of bacteremia occurred during a six-week period in January and February. The development of bacteremia was significantly related to the extent of third-degree burn and to admission to the burn center in January and February but not to the presence of an intravenous cannula, underlying disease, or antimicrobial therapy. E. cloacae was spread by contaminated hands of personnel and by cross-contamination of hydrotherapy water. A shortage of staff appeared to be an important factor in the occurrence of the outbreak. Control measures included an increase in the number of personnel, instruction of personnel in proper aseptic technique, and adoption of a new hydrotherapy protocol.


Marked differences between pigmented and albino rabbits in the concentration of clindamycin in iris and choroid-retina. The concentrations of clindamycin were significantly higher in iris and choroid-retina or pigmented rabbits than in those of albino rabbits after subconjunctival injection. Equilibrium dialysis experiments showed no affinity of clindamycin for synthetic melanin or for collagenase digests of pigmented tissues. In contrast, strips of iris and choroid-retina took up clindamycin rapidly from solution, achieving concentrations substantially higher than those in the medium. Uptake by tissue strips was not influenced by temperature (4 C vs. 37 C), cyanide, or ouabain. However, N-ethylmaleimide, which reacts with sulfhydryl groups, decreased the tissue-to-medium ratios by about 50% for both albino and pigmented choroid-retina. Even in the presence of this inhibitor, the ratio for pigmented tissues remained higher than that for albino specimens. These findings suggest the existence of one, or possibly two, mechanisms of energy-independent accumulation of clindamycin by pigmented ocular tissues: one may relate to protein sulfhydryl bonds that are present in both breeds; the other may involve the pigmentation apparatus.


Levels of exfoliatin antitoxin in pooled human serum globulin. The potential value of human serum globulin for treatment of patients with the staphylococcal scalded-skin syndrome was determined by measuring the exfoliatin antitoxin content of globulin pools. Of 16 lots tested, only four had detectable antitoxin activity, and in these the levels were minimal. These findings suggest that pooled serum globulin is not a practical source of exfoliatin antitoxin.


Effect of chloramphenicol on in vitro function of lymphocytes. The effect of chloramphenicol on the in vitro function of human peripheral blood lymphocytes was studied in assays of lymphocyte transformation and lymphokine production. When lymphocytes were stimulated by phytohemagglutinin, concanavalin A, or pokeweed mitogen in the presence of various concentrations of chloramphenicol, only minimal effects on blastogenesis were noted. However, suppression by chloramphenicol of blastogenesis induced by candida antigen or streptokinase-streptodornase was greater in magnitude and was dose-dependent; blastogenesis was suppressed to 25%--30% or normal levels by concentrations of chloramphenicol of 25--50 microgram/ml. Chloramphenicol had little effect on the production of the lymphokine leukocyte migration inhibition factor by lymphocytes stimulated either by candida antigen or by concanavalin A, whereas puromycin at a concentration of 5 microgram/ml significantly suppressed this response. Thus chloramphenicol appears to suppress antigen-induced lymphocyte blastogenesis significantly but not lymphokine production by stimulated lymphocytes.


Resistance to Listeria monocytogenes in mice: genetic control by genes that are not linked to the H-2 complex. After mice of several inbred strains were injected with Listeria monoyctogenes, two parameters of resistance, the 50% lethal dose and the suppression of bacterial proliferation in spleen, were determined. The strains of mice tested could be segregated into two groups: the resistant C57BL/10Sn mice and the sensitive A/J and DBA/2J mice. Congenic resistant strains of mice were used because they would express the H-2 haplotype of the sensitive strains (H-2a or H-2d) on the background of a resistant strain, C57BL/10Sn. Both the B10.A/SgSn (H-2a) and the B10.D2/Sn (H-2d) mice were as resistant as mice from their background strain and were significantly more resistant than the strains that donated their H-2 locus (A/J or DBA/2J). Therefore, the resistance of mice to Listeria, although genetically controlled, is not controlled by gene (s) linked to the H-2 haplotype. On the other hand, the level of specific immunity to listeria antigens (as indicated by the footpad reaction) was higher in the C57BL/10Sn (H-2b) mice than in either the A/J and B10.A/SgSn (H-2a) mice or the DBA/2J and B10.D2/Sn (H-2d) mice. This observation suggests an H-2 linkage of specific immunity to Listeria.


Infection with Chlamydia trachomatis: involvement of multiple anatomic sites in neonates. In a study of infection due to Chlamydia trachomatis in infants, chlamydiae were recovered not only from the conjunctiva and respiratory tract but also from the vagina and rectum. The timing of recovery suggested that the vagina and conjunctivae are exposed to chlamydiae at birth and that pneumonia and gastrointestinal infection occur later. Sampling of the rectum may be a useful procedure for the diagnosis of chlamydial disease in infants.


Unmasking of actual and potential receptor sites for cholera toxin in intestinal mucosal homogenates. Endongenous and exogenous sialidases appear to unmask sialidase-stable and sialidase-labile gangliosides in intestinal mucosal homogenates by attacking glycoproteins. Exogenous (but not endogenous) sialidase then converts sialidase-labile gangliosides into the cholera toxin-binding, sialidase-stable ganglioside GM1 (galactosyl-N-acetylgalactosaminyl [sialosyl] lactosyl ceramide). Since Vibrio cholerae produces sialidase, these observations may be relevant to the course of cholera.


Defective leukotaxis in monocytes from patients with pulmonary tuberculosis. Because the accumulation of macrophages and their precursors, peripheral blood monocytes, in foci of infection is an important feature of the host reponse to mycobacterial challenge, the leukotactic responsiveness of monocytes from patients with active tuberculosis was evaluated. With a double-filter, in vitro technique, defective leukotaxis was demonstrated in monocytes from 19 of 20 untreated patients, whereas normal leukotactic responses were found in monocytes from 11 of 15 patients with chronic, nontuberculous pulmonary inflammatory diseases. This defect may be related to increased activity of a naturally occurring, heat-stable plasma substance with a molecular mass of approximately 2.3 x 10(5) daltons that inhibited leukotactic responsiveness. Monocyte leukotaxis improved and the leukotactic inhibitory activity of plasma disappeared in most patients while they were on therapy; these phenomena were unrelated to bacteriologic conversion or resolution of symptoms. In vitro studies with isoniazid, ethambutol, and rifampin excluded a direct effect of these drugs or their metabolites on monocytes or on the leukotactic inhibitor in plasma. Thus, defective leukotaxis of monocytes in patients with pulmonary tuberculosis may be an epiphenomenon of the local tissue reaction.


Detection of tobramycin- and netilmicin-induced ototoxicity in guinea pigs with evoked action potentials. To evaluate the action potentials evoked in the cochlea in aminoglycoside-induced ototoxicity, 80 guinea pigs were given 25, 50, 75, or 100 mg of tobramycin or netilmicin/kg per day for 14 or 28 days. Ten other guinea pigs (controls) were given 200 mg of ampicillin/kg per day for 14 or 28 days. Cochlear evoked action potentials (CEAP) before and after treatment were measured, and the cochlea was examined microscopically after treatment. Comparison of initial and final values showed that the threshold of the main negative (N1) wave rose (p less than 0.00001 for dose and duration factors), the amplitude decreased (P less than 0.00001 for dose factors at sound intensities of 120 and 90 dB, P less than 0.001 at 70 dB), and the latency lengthened (P less than 0.0001 for dose factors at 120 and 90 dB). The CEAP method appeared to be more sensitive than microscopic examination of the cochlea for detection of ototoxicity induced by the lower dosages of the aminoglycosides. No significant differences were observed between the effects of tobramycin and netilmicin. In conclusion, the CEAP method appears to be a promising tool for detection of aminoglycoside-induced ototoxicity.


5-fluorocytosine: inhibition of hematopoiesis in vitro and reversal of inhibition by uracil. The in vitro effect of 5-fluorocytosine on human and murine hematopoiesis was studied by means of soft-gel assays for erythroid and myeloid colony-forming cells. The drug consistently inhibited colony formation by granulocyte-monocyte and erythroid precursor cells. Clear effects were observable at concentrations of 5-fluorocytosine of 5 x 10(-5) M (6.5 micrograms/ml), and concentrations of 5 x 10(-4) M (65 micrograms/ml) caused approximately 90% inhibition of cloning of bone marrow myeloid colony-forming cells from mice and normal humans. Greater concentrations of 5-fluorocytosine were required to inhibit erythroid progenitors than to inhibit myeloid precursors. Uracil competitively reversed the toxicity of 5-fluorocytosine. Our data strongly suggest that the hematopoietic toxicity of 5-5-fluorocytosine is mediated through cellular metabolism of the drug. The reversal of mammalian, but not of fungal, cytotoxic effects of 5-fluorocytosine by uracil may have important clinical applications.


Preparation of competent single-cell suspensions of Mycoplasma hominis tets and Mycoplasma salivarium tets for genetic transformation to tetracycline resistance by DNA extracted from Mycoplamsa hominis tetr. DNA extracted from Mycomplasma hominis (Sprott strain), resistant to 100 micrograms of tetracycline/ml transformed M. hominis strain H29 and Mycoplasma salivarium strain S9, which are sensitive to 2.5 and 5.0 micrograms of tetracycline/ml, respectively, to resistance. The transformants were selected on agar medium containing 10 micrograms of tetracycline/ml. Some transformants were resistant also to 20 micrograms of tetracycline/ml, a finding confirming that transformation occurred between homologous and heterologous species and that resistance is stepwise and controlled by several genetic loci. Medium containing 10 micrograms of tetracycline/ml was bacteriostatic. Prototype experiments employing mixtures of strains that were tetr and tets (tetracycline-resistant and tetracycline-sensitive, respectively) demonstrated that tetr mutants and transformants formed typical fried-egg colonies when mixtures containing not more than 10(9) mycoplasmas were spread on tetracycline agar plates. No mutants to tetracycline resistance were detected. Both M. hominis and M. salivarium were competent after treatment with MgCl2 and CaCl2, while Mycoplasma orale type 2 was inactivated. During DNA extraction different quantities of DNA formed insoluble precipitates with protein, thus preventing quantitative experiments.


Synergy of vancomycin plus cefazolin or cephalothin against methicillin-resistance Staphylococcus epidermidis. The in vitro activity of cephalothin, cefazolin, and vancomycin against 25 isolates of methicillin-resistant Staphylococcus epidermidis was determined by means of a broth dilution technique with two sizes of inoculum. The size of the inoculum had a marked effect on the minimal inhibitory concentrations and the minimal bactericidal concentrations of all three antibiotics. With a small inoculum, 100% of the isolates were inhibited by 3.12 micrograms of vancomycin/ml, 76% by 12.5 micrograms of cephalothin/ml, and 64% by 12.5 micrograms of cefazolin/ml. With a large inoculum 100% of the isolates were inhibited by 200 micrograms of vancomycin/ml, 40% by 12.5 micrograms of cephalothin/ml, and 12% by 12.5 micrograms of cefazolin/ml. As determined by a tube dilution checkerboard technique for both sizes of inoculum, the combination of vancomycin plus cephalothin was synergistic against methicillin-resistant S. epidermidis in 45 of 50 cases, and the combination of vancomycin plus cefazolin was synergistic in 39 or 50 cases. These data from in vitro studies suggest that these antibiotic combinations should be evaluated clinically in patients with severe infections caused by methicillin-resistant S. epidermidis.


Non-B hepatitis in Japanese recipients of blood transfusions: clinical and serologic studies after the introduction of laboratory screening of donor blood for hepatitis B surface antigen. Cases of hepatitis virus infection in Japanese recipients of blood transfusions were serologically and clinically analyzed after the introduction of laboratory screening of donor blood for hepatitis B surface antigen by counter immunoelectrophoresis. Non-A, non-B hepatitis occurred in 116 (10.7%) and hepatitis type B in nine (0.9%) of the 1,082 recipients. The incubation period of the post-transfusion non-A, non-B hepatitis cases varied from two to 33 weeks, but most occurred within 15 weeks. In 97 (83.6%) of the 116 cases of non-A, non-B hepatitis studied, the duration of abnormal elevation of the level of serum alanine aminotransferase (glutamic-pyruvic transaminase [SGPT]) was 16 weeks. The cases of non-A, non-B hepatitis could be divided into three groups according to the pattern of elevation of SGPT levels. These findings may suggest either a multiple etiology for non-A, non-B hepatitis or a variety of clinical symptoms with a single etiology for the infection.


Comparison of the features of astroviruses and caliciviruses seen in samples of feces by electron microscopy. Astroviruses and caliciviruses may both be observed in the feces of babies by electron microscopy and are two of the viruses whose relationship to infantile diarrhea is still being established. Although similar in size and in their star-shaped surface configurations, they differ in detailed structure. The astrovirus has a round unbroken edge, a six-pointed star with a white center, and surface hollows that are triangular. The calicivirus has a feathery edge, a six-pointed star with a dark hollow in the center (Star-of-David) appearance), and surface hollows that appear round or oval. Because only a minority of the virions of both viruses exhibit a completely characteristic structure, careful searching may be necessary to distinguish between the two viruses.


Seroepidemiology of infection with hepatitis A and B viruses in an isolated Pacific population. To determine the prevalence of infection with hepatitis A virus and hepatitis B virus in an isolated population, samples of serum were collected from 574 healthy subjects living on the remote Pacific Island of Funafuti. Each specimen was tested for antibody to hepatitis A virus, hepatitis B surface antigen, and antibody to hepatitis B surface antigen by solid-phase radioimmunoassay. Overall, 79.8% of the population showed evidence of previous infection with hepatitis A virus, and 72.5% with hepatitis B virus; the high prevalence of antibody to both viruses in young adults suggested that the majority of infections were acquired in the first decade of life. Although it is known that hepatitis B virus maintains itself in isolated populations through a reservoir of chronic carriers, the reason for the persistently high rate of infection with hepatitis A virus is unknown.


Radioimmunoassay of somatostatin and its application in the study of pancreatic somatostatin secretion in vitro. A sensitive and specific radioimmunoassay for somatostatin is described. With the use of this system, somatostatin release from incubated rat pancreatic islets and perfused rat pancreases has been studied in vitro. Both arginine and glucose, known modulators of insulin and glucagon secretion, were found to stimulate somatostatin release. These results provide additional support for the concept that somatostatin may act as a local regulator of pancreatic A cell function.


Metal-dependent neutral proteoglycanase activity from monolayer-cultured lapine articular chondrocytes. Neutral proteoglycanase and other protease activity from cellular and CM fractions of monolayer-cultured rabbit articular chondrocytes were studied. The cellular fraction comprising soluble cytoplasmic enzymes possessed concentration-dependent elastase-like esterase activity and activity against trypsin and chymotrypsin synthetic substrates but had little caseinase activity. The 20% ammonium sulfate precipitate of CM possessed more neutral caseinase activity than the 60% ammonium sulfate precipitate and the bulk of activity against the synthetic substrates. Activity against bovine nasal septum PG was present in these fractions. Both the 20% and 60% ammonium sulfate fractions reduced the viscosity and the S of the PG substrate. This activity was incompletely inhibited by preincubation with either 5 mM o-phenanthroline or 10 mM EDTA, indicating that it was paritally metal-dependent. The activity in the cellular fraction was also partially inhibited by o-phenanthroline but more so by EDTA. These data indicate that chondrocytes synthesize and secrete into the culture medium neutral proteoglycanase(s) capable of initiating degradation of PG derived from the neutral pH cartilage matrix. The inhibitory profiles, together with recent evidence of enzymes with similar activity extracted from cartilage suggested that the proteoglycanase enzyme(s) may occur in multiple forms.


The effects of dihydrotachysterol therapy on the measurement of plasma 1,25-(OH)2-vitamin D in humans. Anephric patients have undetectable plasma concentrations of 1,25-(OH)2-D, but when anephric patients are treated with DHT2 (Hytakerol), their plasma contains a substance that is co-purified with and displaces authentic 3H-1,25(OH)2-D3 from its intestinal cytoplasmic receptor. The concentration of this substance in the plasma of anephric patients taking DHT2 is proportional to the administered daily dose of DHT2 per kilogram body weight. When DHT2 therapy is discontinued, the substance disappears from plasma with an average t 1/2 of about 8 days.


A method for continuous drug infusion in unrestrained rats: its application in evaluating the toxicity of 5-fluorouracil/thymidine combinations. A method for continuous infusion of drugs into the tail vein of rats was developed. This simple procedure has the advantage of permitting freedom of movement of the animal during extended drug exposure. Utilization of an aluminum sheath as a tail cover prevents destruction of the cannulating apparatus without placing additional stress upon the animal. This method produced no adverse effects upon rats during prolonged infusion (up to 7 days) and may be useful in the routine evaluation of agents having a short plasma half-life. The lethality of FU alone or in combination with TdR was evaluated with this technique. Infusion of up to 10 gm/kg/72 hr of TdR produced no mortality. Co-administration of TdR at 1 gm/kg/72 hr with FU, however, potentiated the toxicity of FU. These data indicate that the toxicity of FU may be modified by provision of TdR with this method.


Blood coagulation system pathophysiology in acute myocardial infarction: the influence of anticoagulant treatment on laboratory findings. Two hundred twenty patients admitted to a Coronary Care Unit were studied by serial plasma fibrinogen chromatography--a method for quantification of HMWFCs, native fibrinogen, and other fibrinogen derivatives in plasma. Enhanced formation of fibrin (intravascular coagulation/thrombosis) is reflected by elevation of plasma HMWFC. One hundred ten patients suffering from documented acute myocardial infarction showed early, sharp elevation of plasma HMWFC (p less than 0.001 when compared to normal and cardiac control groups), a finding which persisted for 10 to 20 days after infarction. Forty-three of the patients did not receive anticoagulant therapy, and the others received either initial heparin, heparin plus warfarin, or werfarin therapy. Plasma fibrinogen chromatographic findings, days 1 to 5, did not differ between the anticoagulated and nonanticoagulated treatment groups, although there were minor differences in the data for days 6 to 10. The results demonstrate that patients with acute myocardial infarction develop a coagulopathy characterized by enhanced fibrin formation, which is influenced to only a minor degree by conventional dosage anticoagulant therapy.


Hypothalamic-pituitary-ovarian axis in uremic women. Menstrual abnormalities are common with decreasing renal function and uremia. Maintenance hemodialysis therapy partially corrects some of the abnormalities. Since the pathophysiology of the menstural irregularities is unknown, the secretion of LH, FSH, estrogen, and PRL was studied under basal conditions and after provocative testing in 13 women undergoing chronic maintenance hemodialysis. Elevated LH concentrations were found in all six premenopausal (44.1 +/- 26.4 mlU/ml, normal follicular phase 4 to 30) and seven postmenopausal (246.1 +/- 171.4 mlU/ml, normal 35 to 200) women studied. FSH values were normal both in the premenopausal women (7.9 +/- 4.1 mlU/ml, normal 2 to 20) and in postmenopausal women (88.0 +/- 69.0 mlU/ml, normal 40 to 200). FSH and LH levels did not show typical ovulatory spikes in three of four women studied. Pulsatile hormonal release was absent. The pituitary response to LH-RH was adequate and prolonged, but the response to clomiphene citrate was variable. The study documents the existence of abnormalities at various levels of the hypothalamic-putuitary-ovarian axis.


Serum alkaline phosphatase in azotemic and hemodialysis osteodystrophy: a study of isoenzyme patterns, their correlation with bone histology, and their changes in response to treatment with 1alphaOHD3 and 1,25(OH)2D3. One hundren seventy-eight azotemic patients, 114 on hemodialysis, had measurements of total serum ALP, and definition of isoenzyme patterns on acrylamide gel electrophoresis. In addition, bone histomorphometry was defined in all of the patients by means of transiliac bone biopsies. Serial estimations over 2 years were carried out on several patients, including some being treated with vitamin D2, 1alphaOHD3, and 1,25(OH)2D3. (1) In both nondialyzed and dialyzed patients, serum ALP showed a significant positive correlation with osteitis fibrosa due to secondary hyperparathyroidism irrespective of the presence or absence of concurrent osteomalacia. Increases in the bone isoenzyme were largely responsible for the rise in total ALP. (2) A higher incidence of osteomalacia (p less than 0.001) was observed in patients on hemodialysis in Newcastle Upon Tyne. In hemodialyzed patients where osteomalacia was accompanied by either no secondary hyperparathyroidism (21 patients) or minimal secondary hyperparathyroidism (14 patients), serum ALP remained within normal limits, giving no indication of the existing osteomalacic bone disease. Isoenzyme studies revealed a high prevalence of the intestinal type and also varied combinations of hepatic, intestinal, and bone types. (3) Good response to vitamin D depended on the presence of significant amounts of the bone isoenzyme. Azotemic osteodystrophy characterized by a raised serum ALP and a prominent bone isoenzyme predicted a good response to vitamin D, and the decrease in serum ALP following vitamin D was the result of a reduction in the bone isoenzyme. Patients with symptomatic dialysis osteomalacic bone disease, accompanied by normal total serum ALP and no elevation of the bone isoenzyme, responded less well.


Clinical studies with a nonvasoactive peritoneal dialysis solution. Topical application of dialysis solution to the rat microcirculation causes a transient vasoconstriction for 2 to 3 min. We assessed the clinical importance of this vasoconstriction by developing a dialysis solution without vasoactive properties, as assessed in the microcirculatory laboratory. The solution was of similar composition to human extracellular fluid. We tested its effects on Cur, Ccr, Cin, and dialysate protein concentration. We found that compared to commercial solutions, the lower osmolality of the NVS resulted in loss of ultrafiltration and decreases in clearance of urea and creatinine. The clearance of inulin was unchanged, and dialysate protein increased, suggesting a major increase in diffusive transport of large solutes. Increased diffusive transport of large solutes with NVS suggests that initial vasoconstriction seen in the rat could be present and clinically important during peritoneal dialysis in humans.


Hb P-Nilotic in association with beta0-thalassemia: cis-mutation of a hemoglobin betaA chain regulatory determinant? Hb P-Nilotic which is produced by a hybrid of beta and delta genes was found in several members of a Sudanese family, three of whom had an associated beta-thalassemia. Chemical analyses confirmed the crossover between positions 22 and 50 of the beta delta P chain. The Hb p-Nilotic heterozygote had completely normal hematology, but the patients with the Hb P-Nilotic--beta-thalassemia condition had moderately severe clinical and hematological abnormalities which were considerably more pronounced than those in the father who had a beta-thalassemia heterozygosity. The absolute cellular contents of normal and abnormal non-alpha chains in these subjects and the results of in vitro chain synthesis analyses suggested that the thalassemia gene in this family is of the beta0 type and that the beta A gene which is present in cis to the beta delta P gene is incapable of being stimulated when the beta0-thalassemia determinant is present in trans. It is proposed that a number of recombination events produced a beta delta P hydrid gene with duplication of the beta A gene in cis as well as a change in an untranscribed strand of DNA which controls the expression of the beta A gene.


Acinic cell carcinoma: a clinicopathologic study of thirty-five cases. A clinicopathologic study of 35 cases of acinic cell carcinomas is presented. Complete follow-up information was available on 31 patients (average period, 7-5 years). The malignant potential of these neoplasms is affirmed by observations that metastases (local and distant) occurred in nine patients and that eight patients died as a consequence of their carcinoma. Retrospective classification of the carcinomas into high and low grade lesions correlated well with ultimate biologic behavior but is unlikely to be successful in an intra-operative (frozen section) mode. This limitation is due to: (a) sampling limitations at the time of primary surgery and (b) the malignant behavior of the occasional low grade carcinoma. Histologic features characterizing high grade carcinomas are local aggressive infiltration and areas of the tumor that appear analogous to the embryonic and post-embryonic terminal tubules and intercalated ducts. The best opportunity for cure of these neoplasms lies in their complete surgical removal at the time of initial treatment. For this, a total parotidectomy is the procedure of choice. Enucleation and local excision is to be condemned.


Interaction of kanamycin and noise exposure. The amount of hair cell damage in the cochleas of guinea pigs after exposure to different experimental conditions suggests that a 'safe' level of kanamycin has a potentiating effect on damage due to noise. Such an effect lasts more than 20 days after the cessation of the drug administration.


Submucous resection. The treatment of choice in the nose-ear distress syndrome. The combination of nasal septal deviation and Eustachian tube dysfunction, in the absence of any other pathology, constitutes the Nose-Ear Distress Syndrome. We have undertaken a clinical assessment of the relationship between uncomplicated deviation of the nasal septum and Eustachian tube dysfunction in Naval Personnel who are serving in environments of primarily increased barometric pressure. 120 candidates to the Submarine, Diving and Aircrew branches of the Royal Navy who presented with the Nose-Ear distress syndrome were initially surveyed. None were able to equilibrate their middle ear pressures at an increased ambient pressure of 3 metres of water. Submucous resection was performed on 116, of whom 110 (94.83 per cent) were able to equilibrate their middle ear pressures at an increased ambient pressure of 9 metres of water post-operatively. Xenon 133 Scintigraphy was performed on a further 25 recruits to delineate the post-nasal airflow. This investigation was performed pre- and post-operatively. Pre-operatively, Scintigraphy showed the presence of turbulence in the post-nasal space, while post-operatively the turbulence was absent. All the candidates were unable to equilibrate their middle ear pressures pre-operatively, but after submucous resection 24 (96 per cent) were able to equilibrate their middle ear pressures at an increased barometric pressure of 9 metres of water.


Complications of long-term nasal and oral endotracheal intubation. A case of multiple complications following long-term endotracheal intubation in an adult is presented. It is important to suspect multiple injuries in such a case and to do a complete radiological and endoscopic examination. In this way various problems will be identified and treated promptly.


The treatment of juvenile multiple laryngeal papillomatosis by suction diathermy. Ten cases of laryngeal papillomatosis treated by suction diathermy are described. The results of this treatment compare favourably with other far more expensive techniques. It is emphasised that proper paediatric anaesthesia is most important.


Recurrence of carcinoma of maxillary antrum after 25 years. A true recurrence of a glandular carcinoma of the antrum was diagnosed 20 years after treatment of the original condition: such an occurrence is rare. Detection of the tumour, however, was delayed as inspection of the original site was not possible during follow-up. Adequate visualization of the tumour area is considered to be of vital importance in the management of cases of malignant disease involving the antrum. The tumour showed a surprisingly good response to radiotherapy.


Dentigerous cyst with oro-antral fistula. A dentigerous cyst of the upper jaw is a common disease, forming 3 per cent of all alveolar cysts. However, its association with an oro-antral fistula is extremely rare and only a few cases have been reported so far.


Mesenchymal chondrosarcomas of the head and neck. Mesenchymal chondrosarcoma, in both osseous and extra-skeletal forms, has a decided predilection for the head and neck region. The two cases presented in this report affirm this tendency and also illustrate the capricious biologic behavior of the neoplasms as manifested by the 30 additional cases recorded in the literature. Histopathologic confusion with hemangiopericytoma is avoided by identification of the cartilage component in the mesenchymal chondrosarcoma and by the undifferentiated nature of the cellular (non-cartilaginous) component.


Cranial nerve involvement in malignant lymphoma. A rare case of malignant lymphomas extensively invading the central nervous system is presented. Cranial nerve involvement is usually a late complication of this disease and its presentation is an ominous sign. This patient presented with symptoms and signs related to the fifth to tenth cranial nerves inclusive. He responded promptly to chemotherapy and has been followed up for 14 months without recurrence.


Primary lymphoma of the parotid gland.

Primary lymphoma of the parotid gland. Primary lymphomas of the salivary glands are very rare. Approximately fifty possible cases have been reported. Whether primary malignant lymphoma of the parotid gland is extranodular or not, is not yet clear. We describe two cases of primary lymphoma of the parotid gland and review the literature of this condition.


Acinic cell carcinoma of intraoral minor salivary gland origin. A case of acinic cell carcinoma involving minor salivary gland tissue is reported. This is an exceptional occurrence. The response to radiotherapy is described and it is suggested that this form of treatment should be considered in these cases.


Acoustic impedance measurements and their importance in cleft palate patients. There is a high incidence of middle ear pathology in cleft palate patients, secretory otitis media being the most common lesion. Impedence audiometry gives more valid information about conditions in the middle ear, as confirmed by exploratory paracenteses. Impedance audiometry, therefore, should be done as a routine, wherever facilities exist, in conjunction with the clinical examination.


The transformation of laryngeal leucoplakia to cancer. In the transformation of laryngeal keratosis to cancer what one may be witnessing is a progression from normal epithelium to keratosis without cellular atypia, through varying degrees of atypia to the stage of carcinoma-in-situ when the full thickness of the epithelium is occupied by atypical cells until finally, with breaching of the basement membrane, an invasive carcinoma occurs (Fig. 12). Although this is, in theory, an attractive sequence, in practice it seems rare to detect an alteration in the epithelium from keratosis without atypia to keratosis with atypia and furthermore most of the carcinomas which developed were not in-situ but invasive. Therefore the significant pathway is probably a change from normal non-keratinizing stratified squamous epithelium via keratosis with atypia to invasive carcinoma (Fig. 13).


Hereditary deafness in children: diagnosis and a family report. A family with hereditary deafness is presented. The mother and ten of her children suffer sensorineural hearing loss. The maternal grandparents and four uncles and aunts were all hard of hearing. Clinical examination revealed no accompanying gross anomalies. In one child, Mondini deformity is the cause of deafness. In the rest of the family isolated membranous labyrinth deformity is suggested. The methods of early diagnosis are discussed, among which polytomography proved the most reliable for diagnosing Mondini and Michel deformities. We demonstrated how early fitting of hearing aids is essential for language acquisition and speech development. Clinical ability to differentiate between subtypes of inner ear hereditary deafness not accompanied by gross anomalies is limited. This problem and the problem arising from late diagnosis are illustrated by a report on a family of ten children, nine of whom suffer sensorineural hearing loss.


Multicentric metachronous primary carcinoma of the upper alimentary tract in 'Biri' smokers. Three Biri smokers with multicentric primary metachronous squamous cell carcinomas of the upper alimentary tract are presented. The susceptibility of the tissues of the upper alimentary tract to multiple malignancies, and the carcinogenic effect of radiation and Biri smoke on these tissues, are discussed. We have tried to assess the role of continuous Biri smoking in the production of a second malignancy of the upper food passages in patients who received radiotherapy for their first tumour.


Internal cysts and fistulae of branchial origin. Cysts and fistulae related to the second and third pharyngeal pouches are described. These anomalies probably occur more often than presumed. Unilateral recurring infections of the tonsillar region, including parapharyngeal abscess, may be caused by anomalies related to the second pouch. Third pouch anomalies are located in the piriform recess. Cysts may reach a size that causes obstruction to the larynx and hypopharynx, while diverticulae cause few symptoms.


Biliary lipid secretion in hypercholesterolemia. A report on the effects of primary bile acid ingestion alone or in combination with plant sterols on serum cholesterol levels, biliary lipid secretion, and bile acid metabolism. Biliary bile acid and cholesterol secretion were measured in four patients with type IIa hypercholesterolemia before and after randomized treatment periods. During these periods either a bile acid mixture (cholic-chenodeoxycholic 2:1, a proportion similar to that endogenously synthesized in health), at a level of 20 mg/kg, or the same mixture plus sitosterols, 200 mg/kg, was fed. Serum cholesterol and the cholesterol saturation of fasting-state bile was also measured. Pretreatment biliary lipid secretion was within normal limits. Bile acid kinetic measurements were also recorded before treatment and showed that cholic acid synthesis was disproportionately decreased relative to that of chenodeoxycholic acid, a finding previously reported by others. Administration of the bile acid mixture increased biliary bile acid secretion in 3 of 4 patients, but did not influence biliary cholesterol secretion. The combination of sitosterol-bile acid, however, caused a relative decrease in cholesterol secretion in bile, and fasting-state bile became unsaturated in all patients. No change in fecal neutral sterol excretion occurred during the beta-sitosterol-bile acid regimen, suggesting that simultaneous bile acid feeding blocks the compensatory increase in cholesterol synthesis known to be induced by beta-sitosterol feeding in hypercholesterolemic patients. Serum cholesterol levels also fell modestly during the sitosterol-bile acid regimen, the decrease averaging 15%. We conclude that the abnormally low rate of bile acid synthesis in patients with type IIa hyperlipoproteinemia does not influence biliary lipid secretion; that increasing the input of the two primary bile acids into the enterohepatic circulation does not increase biliary cholesterol secretion or lower serum cholesterol levels in such patients; and that the usual increase in cholesterol synthesis induced by beta-sitosterol feeding does not occur if bile acids are administered simultaneously.


Synthesis of [24, 25-3H]cholesterol: a new substrate for determining the rate of cholesterol side chain oxidation. A procedure for the synthesis of [24,25-3H]cholesterol from the nonradioactive precursor desmosterol is described. The intermediate, isodesmosterol, which was purified by column chromatography, was formed to protect the original double bond (delta 5-6) from hydrogenation. Tritium was introduced into the side chain by catalytic reduction of the double bond (delta 24-25) of the isodesmosterol in the presence of carrier-free tritium. After ring rearrangement of the iso-[24,25-3H]cholesterol acetate, the acetate was hydrolyzed to form the free labeled cholesterol. Hepatic oxidation of the [24,25-3H]cholesterol side chain release tritium into water which freely equilibrates with cell and body water pools. Thus, the rate of 3H2O appearance corresponds to the rate of cholesterol side chain oxidation. Applications of this method to in vivo, isolated perfused liver, and isolated hepatocyte preparations of the rat are discussed.


Evidence that cholic acid CoA ligase is located asymmetrically on the cytoplasmic surface of hepatic microsomal vesicles. The cholic acid CoA ligase activity of rat liver was quantitatively inactivated by proteolysis with pronase, chymotrypsin, subtilisin, or proteinase K in intact microsomal vesicles. Under the conditions employed, less than 14% of the lumenal mannose-6-phosphate phosphatase activity was lost, and the mannose-6-phosphate phosphatase activity remained highly latent. After microsomal integrity was disrupted with sodium deoxycholate, protease treatment resulted in a loss of greater than 74% of the mannose-6-phosphate phosphatase activity. Cholic acid CoA ligase activity was unaffected by preincubation of microsomes with sodium taurocholate under conditions that led to the complete expression of latent mannose-6-phosphate phosphatase activity. The data suggest that cholic acid CoA ligase activity is located on the cytoplasmic surface of hepatic microsomal vesicles.


Metabolism of potential precursors of chenodeoxycholic acid in cerebrotendinous xanthomatosis (CTX). In patients with cerebrotendinous xanthomatosis (CTX), diminished cholic acid production is associated with incomplete oxidation of the cholesterol side chain and the excretion of C(25)-hydroxy bile alcohols. The aims of this investigation were 1) to provide quantitative information on the pool size and production rate of chenodeoxycholic acid by the isotope dilution technique; and 2) to investigate the possible existence of a block in chenodeoxycholic acid synthesis and explain the absence of chenodeoxycholic acid precursors in CTX. After the injection of [24-(14)C]chenodeoxycholic acid, measurements of chenodeoxycholic acid pool size and production rate in a CTX subject were, respectively, 1/20 and 1/6 as great as controls. Further, three potential precursors of chenodeoxycholic acid, namely [G-(3)H]7alpha-hydroxy-4-cholesten-3-one, [G-(3)H]5beta-cholestane-3alpha,7alpha,25-triol, and [G-(3)H]5beta-cholestane-3alpha,7alpha,26-triol, were administered to the CTX and control subjects and the specific activity curves of [G-(3)H]cholic acid and [G-(3)H]chenodeoxycholic acid were constructed and compared. In the control subjects, the two bile acids decayed exponentially, but in the CTX patient maximum specific activities were abnormally delayed, indicating the hindered transformation of precursor into bile acid. These results show that chenodeoxycholic acid synthesis is small in CTX and that the conversion of 7alpha-hydroxy-4-cholesten-3-one, 5beta-cholestane-3alpha,7alpha,25-triol, and 5beta-cholestane-3alpha,7alpha,26-triol to both chenodeoxycholic acid and cholic acid were similarly impaired.


Improved methods for the solubilization and assay of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase. A method for solubilizing HMG-CoA reductase is described that reproducibly yielded approximately 190% of the activity assayed in rat liver microsomes. Optimal solubilization occurred when microsomal membranes were frozen at a fixed concentration, thawed, homogenized in a buffer containing 50% glycerol, and incubated at 37 degrees C for 60 minutes. A rapid spectrophotometric assay of the reductase has been developed and the optimal conditions defined. Using this assay, the kinetics were determined for HMG-CoA reductase purified to a specific activity of 17,400 nmol NADPH oxidized per minute per mg protein.


Quantitative assay of conjugated and free bile acids as heptafluorobutyrate derivatives by gas-liquid  chromatography. Quantitative analyses of individual bile acids in biological samples are limited by the lengthy multistep preparations necessary. Using heptafluorobutyric acid anhydride in pyridine as derivatizing agent, we reduced several steps to one. Bile acids and their glycine and taurine conjugates form stable heptafluorobutyrate derivatives, climinating the need for deconjugation and preparation of methyl esters. The derivatives have been characterized by mass spectrometry, and optimum reaction yields have been determined. Operating conditions for analyzing the bile acid heptafluorobutyrates by gas-liquid chromatography on various column packings were investigated, and 0.5% QF-1 or 3% OV-255 was found suitable. The bile acid derivatives were identical whether starting with the bile acid or the glycine or taurine conjugates. The procedure was applied to a quantitative analysis of artificial mixtures of bile acids and bile conjugates, and also of human bile. The results compared favorably to those obtained with a 3 alpha- and 7 alpha-hydroxysteroid dehydrogenase fluorimetric method.


Mechanism of insulin resistance in adipocytes of rats fed a high-fat diet. Insulin's ability to stimulate glucose metabolism is severely diminished in the adipose tissue of rats fed a high-fat diet as compared to that of rats fed a low-fat diet. To elucidate the mechanism for this effect we have measured the binding of insulin, the hormone effect on 2-deoxyglucose uptake and the major pathways of [1-(14)C]glucose metabolism, and the activity of lipogenesis-related enzymes in adipocytes of rats fed a low- or high-fat diet for 7 days. Rats fed high- or low-fat diets bound equal amounts of insulin per adipocyte at all insulin concentrations tested. Basal and maximally insulin-stimulated 2-deoxyglucose uptake per fat cell were reduced in high-fat-fed rats. However, the insulin stimulation over basal was the same in both groups (230%). Submaximal doses of insulin produced equivalent increments of 2-deoxyglucose uptake in both groups, as would be predicted by the binding studies. The effect of both submaximal and maximal insulin concentration on the labeling of CO(2) and fatty acids was markedly decreased by high-fat feeding. The insulin response of the glyceride-glycerol pathway was less severely, though significantly, reduced. Acetyl CoA carboxylase and malic enzyme in adipocytes of high-fat-fed rats were reduced to 13% of the activity in the low-fat-fed rats. Glucose-6-phosphate and 6-phosphogluconate dehydrogenases were decreased to 20% and 34% of their activities in low-fat-fed rats, respectively. These reductions paralleled the changes in insulin-stimulated glucose oxidation and fatty acid synthesis. The data therefore strongly suggest that the blunted response of glucose metabolism to insulin in adipocytes of high-fat-fed rats is a result of a decreased intracellular capacity to utilize glucose for lipogenesis.


Analysis and quantitation of free ceramide containing nonhydroxy and 2-hydroxy fatty acids, and phytosphingosine by high-performance liquid chromatography. Reaction of ceramides containing nonhydroxy fatty acids with benzoyl chloride in pyridine at 70 degrees C for 1 hr resulted in N-benzoylation to form N,N-acyl,benzoyl derivatives; O-benzoylation also occurred. However with ceramides containing 2-hydroxy fatty acids and phytosphingosine only O-benzoylation occurred even on prolonged treatment. Only O-benzoylation occurred on reaction with benzoic an hydride. However, the benzoylation of ceramides with phytosphingosine could not be achieved with benzoic anhydride and this benzoylation was performed by reaction with benzoyl chloride at 70 degrees C for 4 hr. Because N,N-acyl,benzoyl derivatives of ceramides containing nonhydroxy fatty acids produced by treatment with benzoyl chloride overlap methyl benzoate on high-performance liquid chromatography, benzoic anhydride was preferable for benzoylation of ceramides with nonhydroxy and 2-hydroxy fatty acids. On the other hand, the reaction with benzoyl chloride at 70 degrees C for 4 hr was used for quantitation of benzoylated ceramides containing 2-hydroxy fatty acids and phytosphingosine. 3-(p-Phenylbenzoyl)estrone was used as an internal standard for both reactions and values for ceramides containing 2-hydroxy fatty acids obtained by the two reactions were in good agreement. This procedure was applied to measurement of the ceramide levels in the brain, liver, and kidney of rats during development. The levels of ceramides containing nonhydroxy and 2-hydroxy fatty acids in the brain, liver, and kidney increased to the adult levels and then remained unchanged. Ceramide with phytosphingosine was detected in the liver and kidney, where its concentration gradually increased with age, but it was not found in the brain. The composition of nonhydroxy fatty acids were also analyzed.