Effects of methylphenidate alone and in combination with behavior modification procedures on the behavior and academic performance of hyperactive children. Twenty hyperactive 6- to 9-year-old children of normal intelligence were studied in a half-day laboratory classroom in a 2-week period baseline-treatment-reversal design for behavior modification. Under double-blind conditions half the children were placed on .3 mg/kg of Ritalin and half on placebo for the entire program. The classroom program consisted of a group period with immediate reinforcement possible, and an individual time period without immediate reinforcement possible. Behavior modification caused a significant decrease in nonattending, out-of-seat, inappropriate vocalizing and inappropriate peer interaction behavior in the group period. Fidgeting, a nontargeted behavior, was not significantly decreased during this period but did significantly decrease as a result of medication. No other drug effects occurred during this period. During the individual period, the results were essentially reversed. There were no significant behavior modification effects observed. Significant reductions resulted from medication in all behaviors except out-of-seat and fidgeting. Behavior modification alone significantly affected the two academic measures. No signigicant effects were seen on the Conners Abbreviated Teacher Rating Scale. No significant interactions were noted between medication and behavior modification.


Span of apprehension in hyperactive boys. The spans of apprehension of hyperactive and normal boys were compared with a forced-choice letter-recognition task developed by Estes (1965). This task provides an estimate of the span which is relatively insensitive to either memory or motivational influences. The span size was the same for both groups when visual "noise" was absent. In the presence of noise, span size for the hyperactive boys decrease. Moreover, as the number of noise letters increased, the reduction in span size observed for the hyperactive boys increased. It is argued that this increasing reduction in span size represents a true deficit in attention.


Dextroamphetamine and placebo practice effects on selective attention in hyperactive children. Three groups of boys referred to a hospital study unit for evaluation of hyperactive behavior were tested on a classification task involving selective attention while on either dextroamphetamine (D) or placebo (P). In two sessions, groups had D first, P second (DP), or PD, or PP. Amphetamine reduces a response times in general and reduces interference due to orthogonally varying irrelevant information. Practice while on placebo improves performance in a subsequent placebo session. Practice while on amphetamine does not, however, improve performance in the subsequent session on placebo. Assessment of the extent of the drug-state-related practice effect is necessary for evaluation of long-term benefits of dextroamphetamine therapy in these children.


Continuities in maternal reports and child behaviors over time in hyperactive and comparison groups. Maternal reports, observations of nursery and elementary school behavior, and teacher ratings of problems were available for hyperactive and control children who had participated in a longitudinal study. This paper examines consistencies in maternal reports and child behaviors over time, and their relationship to teacher ratings in elementary school. Maternal reports of infant sleep difficulties were related to maternal ratings of hyperactivity at 4 1/2 and 6 1/2 years. Maternal ratings of activity at 4 1/2 were predictive of 6 1/2-year ratings of hyperactivity and conduct problems. In addition, behavior in a research nursery at 4 1/2 predicted teacher ratings of problems and classroom behavior in grade two. Hyperactive preschoolers who left the table most during structured activities were more often out-of-seat and off-task in school. Controls who were more aggressive in the nursery were more disruptive in the classroom. These data indicate continuities in both maternal reports and child behaviors.


Formal and informal classroom settings: effects on hyperactivity. The behavior of hyperactive children was compared to controls in two different classroom settings. One setting (Informal) involved choice and variety of tasks; the other setting (Formal) involved teacher specification of a small number of tasks. As assessed by a composite observational measure of hyperactivity, there were significant differences between the hyperactive and control groups in the Formal but not in the Informal setting. Analyses of five individual categories of hyperactive behavior showed that, with one exception, the hyperactive group tended to display higher frequencies of behavior than did controls in both settings. For two of the categories, the difference between the groups was significantly larger in the Formal than in the Informal setting. Finally, a modified observational code was suggested that differentiated hyperactives from controls equally in the two settings.


The reaction of adolescent offenders and nonoffenders to nonverbal emotion displays. Adolescent male offenders and nonoffenders were compared on two tasks designed to assess reactions to the nonverbal emotion expressions of others. It was found that the offender group was less accurate in labeling another's emotion states but that this difference was apparently a function of verbal intelligence. Further, the results of the second task, a task not related to verbal intelligence, indicated that the punitive decisions of both the offender and nonoffender groups were similarly affected by the actors' nonverbal emotions. These results wree discussed as indicating that much of the social insensitivity often ascribed to delinquents may simply be a result of impoverished verbal skills among this group. A deficit in verbal skills may preclude delinquents from adequately describing their perceptions of the emotional reactions of others and it may also necessitate a nonverbal, rather than verbal, reaction to others' emotions.


The children of Mexican-American migrant workers: a population at risk? The present study assessed intellectual, academic, and emotional strengths and weaknesses for a group of Mexican-American children of migrant farm workers. In order to test the vulnerability hypothesis, the test profiles of these children were contrasted with those of two groups of black children with similar demographic makeup. The children of one group (Clinical Black) had been referred previously for a psychological consultation while the children of the other (Nonreferred Black) had not. Across dependent measures, between-groups contrasts tended to describe the scores of the Migrant children as similar to those of the Clinical Black children and significantly below those of the Nonreferred Black children. The findings suggested specific intellectual, academic, and emotional vulnerabilities of the Migrant children and demonstrated the need to develop ameliorative programs for these children.


Monocyte cellular function in asthmatic patients on alternate-day steroid therapy. Monocyte cellular function in 15 asthmatics on alternate-day steroid therapy (mean dose of prednisone, 45.4 +/- 17.45 mg qod) was studied at 8 A.M. and noon after receiving an 8 A.M. steroid dose and at 8 A.M. the following day, and was contrasted to function in 16 healthy controls. Monocyte chemotaxis, bacterial killing and phagocytosis, and oil phagocytosis were not significantly altered by the steroid dose. On the other hand, all the patients experienced monocytopenia, lymphopenia, and neutrophilia 4 hr following the administration of steroid. The lack of functional impairment on this clinically relevant steroid regimen is consistent with the lack of serious infections seen in patients on such regimens. This study re-enforces the need to differentiate the effect of even small doses of steroid on circulating cell populations from direct effects on cell function which occur only with very high or frequent steroid dose regimens.


The safety of immunotherapy during pregnancy. A group of 121 pregnancies from 90 atopic mothers who had received immunotherapy during pregnancy were studied retrospectively with the use of physician or maternal questionnaires and obstetric records. The incidence of prematurity, toxemia, abortion, neonatal death, and congenital malformation was no greater than that for the general population. The offspring of the treated mothers developed allergic disease as frequently as children born into allergic families. A group of 147 untreated pregnancies of atopic mothers were similarly studied, and the incidence of those parameters were similar except for a greater incidence of abortion. The implications of these findings are discussed.


Localized heat urticaria.

Localized heat urticaria. The pathophysiology of localized heat urticaria was studied by performing a heat challenge on a patient with this disease. Serum levels of total hemolytic complement, C3, and factor B decreased following heat challenge, whereas levels of C4 and C5 did not. Plasma histamine levels remained unchanged. Electron microscopic studies of affected tissue revealed endothelial cell damage and neutrophilic degranulation in the affected area. Mast cells remained intact. These data imply that activation of the alternative complement pathway is involved in the pathogenesis of localized heat urticaria and that mast cell histamine release does not play a significant role in this disease.


Preschoolers on alternate life-style diets. Associations between size and dietary indexes with diets limited in types of animal foods. One hundred nineteen vegetarian preschool children whose parents volunteered were studied in their homes. Length, weight, triceps and subscapular skinfolds, and head circumference were measured. Information on extensiveness of avoidances of animal food groups, parental dietary group, birth weight, parental heights, and the child's status with respect to breast feeding at the time of measurement was obtained by parental report. All infants under six months old were breast fed. More of the children's measurements were below the Harvard 50th percentiles for length and weight than would have been expected after six months of age, but not until then. Vegetarian children were also leaner (triceps and subscapular skinfolds) after six months of age than would be expected from Tanner-Whitehouse standards. Head circumferences were similar to standards and within normal limits at all ages. An association between smallness, lightness, and leanness was found among the vegetarians, with these characteristics being more pronounced among the older, non-breast fed, macrobiotic children on diets with limited variety in terms of animal food. While smallness and low weight-for-length were associated with leanness, largeness and high weight-for-length did not imply obesity in these subjects.


Effects of high- and low-fiber diets on human feces. Increased- and low-fiber diets composed of common hospital foods were tested for their ability to alter stool weight, frequency of defecation, transit time, and absorption efficiency. The diets were calculated to be isocaloric and similar in protein, fat, and carbohydrate. They were dissimilar in ash and fiber. No stimulants, secretagogues, food concentrates, or extracts were used on either diet. Six normal, healthy, male subjects consumed each constant diet for five to eight days. Diets and stool composites were analyzed for protein, fat, fiber, moisture, and ash. Carbohydrate and calories were calculated. Stool weight during ingestion of the low-fiber diet averaged 51 gm. per day; subjects had an average of one bowel movement every 33 hr.; and the mean transit rate was 48 hr. When subjects consumed the increased-fiber diet, values were 157 gm., 19h., and 12 hr., respectively. Measurements of fecal protein, fat, carbohydrate, and calories from the low-fiber diet were less than half those observed when the increased-fiber diet was consumed. By altering specific components of a normal diet, e.g., fiber, significant differences can be made in bowel habit and absorption efficiency.


Improving dietary education for patients with hyperlipidemia. In this study, an attempt was made to analyze the effectiveness of a health communication campaign in bringing about dietary changes in persons with hyperlipoproteinemia. The author developed an instructional booklet for each of three types of hyperlipidemia. Sixty-eight patients identified by cooperating physicians were provided copies of the booklet (a mass media message) or a booklet combined with a telephone consultation (interpersonal communication). Fifty-six persons returned a mail questionnaire and indicated that the instructional technique was successful in changing both attitude and behavior. The positive correlation between changes in attitude and behavior suggests that a change in attitude about food would be likely to bring about alterations in eating habits. Questionnaire replies also demonstrated that the participants responded favorably to presentation techniques emphasizing the characteristics of simplicity, compatibility, trialability, and relative advantage. The combination of a telephone consultation plus a mass media message was significantly related to a greater change in attitude. These results have implications for the future development of instructional materials, as well as for the provision of dietary consultation by persons with a limited opportunity for personal contact with patients.


Nutritional status of main food preparers and of nutrition education assistants. The dietary habits and nutritional status of nutrition education assistants (NEAS) in the Expanded Food and Nutrition Education Program in Missouri were compared with a representative population of Missouri main food preparers (MFPs) from the same regions of the state. Dietary histories indicated that the NEAs tended to have better habits than the MFPs. Using biochemical parameters for determining status for iron, vitamin A, ascorbic acid, and riboflavin, the differences between the two groups were not as great as for the dietary intakes. However, almost half of the MFPs were taking at least one vitamin or mineral supplement compared with 17 per cent of the NEAs. A low dietary intake, coupled with some anemia, probably relating to a low iron intake, was a major nutritional problem for both groups of subjects. Low plasma vitamin A values were also observed among a large proportion of both groups, and obesity was a widespread problem for both groups.


Hormones: beneficial or dangerous to the aged? Postmenopausal women lose 75-85 percent of estrogen and progesterone production compared with that in their reproductive years. The marked decrease in estrogen results in changes in the uterus, cervix, vaginal lining, glandular breast tissue, and the rate at which bone loses thickness. Some of these changes result in symptoms which range from unpleasant to disabling. Many of the symptoms can be alleviated by treatment with estrogens. However, recent reports have been linking estrogen usage with carcinoma of the uterus. Postmenopausal women should be treated with estrogens when the situation demands, but the dosage should be kept as low as possible and progestational agents should also be administered.


Interest in geriatrics at a university Department of Medicine. To estimate available resources for teaching geriatrics to medical students, a survey of the members of a university Department of Medicine was undertaken. Information was elicited in six areas: 1) personal data, 2) characteristics of each physician's clinical practice, 3) potential interest in a Division of Geriatrics, 4) the comparative levels of aggressiveness indicated in the management of an old patient versus a young one, 5) the attitude of physicians toward the elderly, and 6) a rough measure of knowledge of the demographic characteristics of the elderly. The results suggest that a significant number of faculty members of a university medical school would indeed be interested in teaching and research in the field of geriatrics.


Training objectives of a well developed geriatrics program. The Department of Family Practice and Community Health of the University of Minnesota Medical School is composed of six affiliated units. Each unit is allied with a hospital complex and operates an independent family practice clinic. Since each unit has unique demographic characteristics, general objectives for training residents in geriatric medicine have been developed. Cognitive, attitudinal and skill objectives also have been identified. The unit director implements the general objectives by using the special strengths of that particular unit.


A concept of loneliness in the elderly.

A concept of loneliness in the elderly. The author's concept of loneliness is described, especially as it concerns the elderly. An attempt is made to develop an understanding of the word, with its implications for guidance in the nursing profession. The nurse's defenses against her own feelings of loneliness and depression can inhibit therapeutic interactions with lonely or depressed patients. Appropriate nursing action depends upon the knowledge, correlation and use of many other concepts such as those related to dynamics of behavior, personality structure and development, or problem solving. Practical recommendations for nursing intervention are outlined. Emphasized is the necessity for the nurse to understand the concept of loneliness so that she may render effective service.


Nurses' notes on nursing home patients: are they effective? The notes written by nurses in a level-III nursing home were reviewed in detail. They were found to be neither pertinent nor meaningful in most cases. The daily writing of these notes required a large percentage of the nurses' time but they were seldom, if ever, read by physicians. An alternate system of writing notes on a weekly basis is suggested.


Fate of parathion in artificially fortified grape juice processed into wine. "Semellon" grape juice fortified with a high level of 25 ppm parathion was fermented using Saccharomyces cerevisiae var. ellipsoideus. After 12 days inte parathion levels in the wine and lees were 10.3 and 156 ppm, respectively; the paraoxon, aminoparathion, and p-nitrophenol levels in the wine were 0.16, 0.20, and 4.5 ppm, respectively, and in the lees were 0.04, 3.1 and 10 ppm, respectively. Thus, hydrolysis of parathion to p-nitrophenol and parathion sorption to sedimented particulate matter were important pathways for parathion residue reduction in the wine. The 56-day-old finished wine just prior to bottling contained 8.8 ppm parathion, 0.04 ppm paraoxon, 0.21 ppm aminoparathion, and 3.0 ppm p-nitrophenol. Two months storage at 24 degrees, 12 degrees, 4 degrees, and -20 degrees C had no effect on paraoxon and aminoparathion residue levels in the wine; parathion residues in wine decreased at all storage temperatures.


Persistence of residues in water and sediment of a fresh-water lake after surface application of technical chlordane. A fresh-water lake, free from detectable pesticide residues before this study, was treated with a commercial formulation of technical chlordane. Water and sediment samples were analysed for chlordane residues 7, 24, 52, 279. and 421 days after treatment. Residues moved rapidly from the water to the lake bottom, supporting earlier results of a laboratory study with other organochlorines. In water, alpha- and gamma-chlordane concentrations remained proportional to total chlordane concentrations, as determined by total-peak area measurements of gas-liquid chromatogrammes. But in bottom sediments, alpha- and gamma-chlordane were more persistent than other constituents of technical chlordane, supporting recent evidence that quantification of technical chlordane residues on the basis of gamma-chlordane quantification only (or that of alpha- and gamma-chlordane only), can lead to incorrect results and that this method, although simple and fast, should no longer be recommended.


Polydactyly of the thumb: abnormal anatomy and treatment. In nine years 26 duplicated thumbs were seen in the hand clinic of a hospital for crippled children. Using a classification based on the level and degree of duplication and a knowledge of the abnormal anatomy, reconstruction was done. This surgery was done with proper skin incisions, reconstructions of the collateral ligaments, centralization of the flexor and extensor tendons, and alignment of the bones and joints by corrective osteotomies. Simple ablation or incomplete correction produced complications such as skin contractures, increased angular deformities, unstable joints, and intrinsic weakness. In all of the patients, the appearance was improved, opposition was maintained, and postoperative function was not impaired.


Osteochondritis dissecans and brachymesophalangia: a hereditary syndrome. A kindred with an unusual congenital hand and foot anomaly is presented. It is an autosomal dominant hereditary disorder characterized by brachymesophalangia, slight clinodactyly, deformed metacarpal heads, osteochondritis dissecans in metacarpophalangeal joints, absence or hypoplasia of the ulnar styloid processes, and brachyhypophalangia in the feet.


Common variations of the radial writs extensors. An anatomical study of the extensor carpi radialis longus (ECRL) and brevis (ECRB) in 173 upper limbs demonstrated abnormalities in 50%. Interconnecting tendons between the longus and brevis were found in 35% of limbs, the most common variation (26%) being a tendon arising proximally from the ECRL and inserting distally with the ECRB. In 42 limbs (24%) an extra muscle (the extensor carpi radialis intermedius) was present; many were large enough to activate a tendon transfer. Two case reports illustrate the use of extra muscles in the surgical rehabilitation of patients with paralytic disorders.


Bilateral extraosseous chondromas of the hand in a patient with chronic renal failure. Extraosseous cartilage tumors of the hand are rare. Histologically they can be distinguished from chondrosarcomas. We report a patient with bilateral extraosseous chondromas of the hand with rapid appearance and growth of this unusual tumor during chronic renal failure. The clinical and histological characteristics of these benign tumors are reviewed, and the possible role of the patient's depressed cellular immunity in the appearance of these chondromas is discussed.


Chronic fracture-subluxation of the proximal interphalangeal joint. Reported methods of treatment of chronic fracture-subluxations of the proximal interphalangeal joint require wide dissection and prolonged immobilization and often result in limited motion. Open reduction with minimal dissection, immobilization for 11 days to allow soft-tissue healing, and the early institution of active flexion exercises protected by an extension block splint was done in four patients. The long-term results demonstrate restoration of a stable joint with satisfactory motion and minimal subjective complaints.


Results following removal of silicone trapezium metacarpal implants. In 100 consecutive patients treated by arthroplasty using Swanson's trapeziometacarpal implant, 32 showed subluxation or dislocation of the implant. Although function was excellent in those with a correctly placed implant, 18 of the 32 had disturbances of function and complaints which necessitated removal of the implant. An operative technique is described for removal of the implant and interposition of the imbricated abductor tendon. The result after removal of the Swanson implant was better than that following primary excision of the trapezium, especially with regard to stability and strength. Thus satisfactory function may be retrieved in patients in whom the implant has dislocated.


Percutaneous recordings from peripheral nerves (preliminary communication). The recently developed technique provides a precise assessment of neutral activity via intrafascicular axonal recordings from human peripheral nerves. With this technique it is possible to record from cutaneous afferent fibers. Receptive fields belonging to cutaneous afferents are readily perceived by patients due to orthodromic conduction of activity induced by mechanical activation of appropriate fibers. Temporal synchrony between peripherally applied stimuli and the proximally placed recording electrode assure continuity of the nerve. By sampling axonal activity from cutaneous fasciles at regular intervals following trauma of undefined etiology, a definite appraisal can be made concerning the integrity of a nerve and, hence, the need for surgical intervention. This is a preliminary report of the first such applications of this technique.


Closed system venography in the evaluation of upper extremity hemangiomas. Closed system venography is a technically simple radiographic technique of minimal patient morbidity and inconvenience that has proven to be useful in evaluating the extent of upper extremity hemangiomata and in planning its treatment. The only apparent contraindication to the technique is allergy to the radiopaque dye. In a recent series of cases, this technique of venography has yielded more information than has arteriography.


The lupus hand: a new surgical approach. Three patients with lupus erythematosus developed severe hand deformities which, over the course of a year, went on to fixed volar subluxation of the proximal phalanges with some ulnar drift. The flexion deformity at the metacarpophalangeal joints of nearly 90 degrees was not correctable passively, and the palmar skin became macerated. The articular cartilage of the metacarpal heads and proximal phalanges was well preserved and replacement arthroplasty was not justified. Contracture not only in the intrinsics but also in the long flexors was not relieved by operation on the soft tissues. A step-cut metacarpal shortening did correct the remaining intrinsic and long flexor contracture, corrected the volar dislocation, restored full metacarpophalangeal extension, and maintained a full range of flexion. Position of the bones was maintained by intramedullary Steinmann pins and crossed Kirschner wires and, in one patient, by a small screw. No recurrences developed in the follow up period of 18 to 96 months.


Bowen's disease of the nail bed: a case presentation and review of the literature. Since 1919, when Bowen first described intraepidermal squamous cell carcinoma of the nail bed, only 11 cases of this rare disease have been reported in the literature. The 12th case of Bowen's disease, presented here, offers an alternative method of treatment using 5-fluorouracil combined with a keratolytic ointment.


An apparatus to assess function of the hand. An apparatus was developed to study the movement, strength, and manipulative ability of the hand. To obtain normal values, measurements were made on 80 women and 65 men between the ages of 17 and 70 years. As a comparison, 30 female patients with severe rheumatoid arthritis were studied. There was little difference in the motion between men and women and among the different age groups. In pinch strength men were 40 percent stronger than women, but in grip strength men were twice as strong. In manipulative ability, there was a decline with age. The arthritic patients were compromised severly in both active motion and strength. In the latter they were between one third and one tenth of normal. Manipulative ability was one half of normal. A measurement system, such as described, could be used for the evaluation of results of various operative procedures.


Improved intravenous regional anesthesia for surgery of the hand, wrist, and forearm. The second wrap technique. In 330 consecutive anesthetics administered over a period of 24 months, an improved method of upper extremity intravenous regional anesthesia, entitled "the second wrap technique," included wrapping the extremity a second time with a Martin rubber bandage after the extremity was prepared and draped. In addition, a Penrose drain tourniquet often was applied during injection of the 0.5% lidocaine. No complications occurred. The technique provides a nearly bloodless operative field, improves the anesthesia, diminishes tourniquet pain, lessens the contraindications, and requires no premedication. The only contraindications are allergy to lidocaine, infection, operating time over 2 hours, and severe hypertension.


Case report: false aneurysm of the hand secondary to digital amputation. A false aneurysm in an amputation stump of the second metacarpal produced a pulsatile mass and throbbing pain. Operative findings disclosed a false aneurysm arising from the digital artery on the radial side of the middle finger, with entrapment of the nerve to the ulnar side of the index digit within the aneurysmal sac. Excision of the aneurysm, relocation of the traumatic neuromas, and amputation of the index ray relieved the symptoms.


Coloidal gold, ferritin and peroxidase as markers for electron microscopic double labeling lectin techniques. Three markers, colloidal gold, ferritin and peroxidase, were checked for usefulness in double labeling of lectin-binding sites. The amount of various lectins for the stabilization of good sols of a different particle size was evaluated. Several lectin-gold complexes were prepared for electron microscopic labeling purposes, and the optimal amount of various lectins needed for stabilization of gold solutions of a different particle size was determined. The following combinations were investigated for their usefulness in labeling two different lectin-binding sites: lectin-gold and lectin-gold (different particle size), lectin-gold and lectin-ferritin, as well as lectin-ferritin and lectin-peroxidase. Of these combinations the latter did not give satisfactory results for double labeling. In all single and double labeling techniques with the above mentioned markers the quantitative evaluation of the number of lectin-binding sites is not feasible, but these techniques will be of considerable value for the investigation of the dynamics of different lectin-binding sites on the cell surface.


A histochemical method for the identification of 9-O-acyl sialic acid. An investigation of bovine submaxillary gland and intestinal mucins. Prolongation of the initial periodate oxidation step of the periodate-borohydride technique/potassium hydroxide/periodic acid-Schiff and periodic acid-Thionin Schiff/potassium hydroxide/periodic acid-Schiff sequences produced little or no change in the diagnostic staining for the potassium hydroxide/periodic acid-Schiff effect, exhibited by the colonic epithelial mucins of man and rat and the Brunner's gland mucin of rabbits. In contrast, there was a gradual, but clear decrease in the intensity of such staining of bovine submaxillary gland mucins. It was concluded that, in the intestinal mucins studied the potassium hydroxide/periodic acid-Schiff effect was due to sialic acids bearing O-acyl substitutents at positions C7 and/or C8 whereas in bovine submaxillary gland mucin the potassium hydroxide/periodic acid-Schiff effect is probably due, at least in part, to the presence of 9-O-acyl sialic acids. This investigation has led to the development of a technique which can be used to identify 9-O-acyl sialic acids.


Acid D-ephedrine-O-phosphatase activity in rate ventral prostate lysosomes. Hydrolysis of D-ephedrine-O-phosphate (DEP) by rat ventral prostrate was investigated by electron microscopic hystochemistry. In contrast to previous reports that DEP is exclusitely hydrolyzed by a secreted nonlysosomal acid phosphatase, enzyme reaction product was present in supranuclear and basal lysosomes.


The use of the fluorescent probe 8-anilinonaphthalene sulfate (ANS) for collagen and elastin histochemistry. The reactivity of skin elastin and collagen to ANS (8-anilinonaphthalene-1-sulphonate) dissolved in nonpolar (e.g. butanol) and polar (distilled water, phosphate buffer at pH 2.8 and 8.2) solvents was compared in histological sections. The elastin fibers were demonstrated to differ from collagen at the fluorescence microscopy level under certain ANS staining conditions. When treated with butanol-ANS solutions, both fiber types exhibit a blue fluroescence with spectral emission profiles alike. However, the elastin emission values in fluorescence-relative units are larger than those exhibited by collagen. On the other hand, for fibers treated with ANS dissolved in polar solvents, elastin and collagen differ in terms of wavelengths of their emission maxima. The ANS attachment to substrate is ascribed to be predominantly nonpolar for dye butanol solutions. When the dye is dissolved in polar solvents, however, polar bindings between ANS and the stained substrate may be established that contribute to emission maximum shift to longer wavelengths.


Bacterial contamination in a modern operating suite. 3. Importance of floor contamination as a source of airborne bacteria. The redispersal factor for bacteria-carrying particles from a contaminated floor was determined after mopping, blowing and walking activity. Walking gave the highest redispersal factor, 3.5 X 10(-3) m-1, which was three times higher than for blowing and 17 times higher than for mopping. The mean die-away rate for the bacteria-carrying particles used was 1.9/h without ventilation and 14.3/h with ventilation. It was calculated that in the operating rooms less than 15% of the bacteria found in the air were redispersed floor bacteria.


Bacterial contamination in a modern operating suite. 4. Bacterial contamination of clothes worn in the suite. Clean clothes in the staff dressing rooms were heavily contaminated with bacteria, mainly Bacillus sp., but Staphylococcus aureus were found on 14% and Clostridium sp. on 10% of the garments examined. A comparison of the occurrence of Staph. aureus on shirts worn by staff in wards and operating departments showed ward shirts to be contaminated more heavily and with more strains. Examination of sterile gowns worn by surgeons showed that 70% were contaminated with Staph. aureus after operation. Of the strains isolated 31% were identical with those carried by the surgeon or by the patient operated on, but for the remainder no source could be found.


Studies on Rift Valley fever in some African murids (Rodentia: Muridae). Brains, spleens and livers of 2214 murids, 27 shrews and 7 dormice, trapped at 7 sites in Rhodesia, were tested in 277 pools for the presence of Rift Valley Fever virus. There were no isolations of Rift Valley Fever, but 69 isolations of an unidentified virus were obtained. Sixteen out of 867 sera had low-titre haemagglutination-inhibition activity against Rift Valley Fever antigen, but only one out of 1260 sera had neutralizing antibody. The evidence suggests that murids fail to encounter infection in nature and are unlikely to play a role in circulation and dissemination of Rift Valley Fever virus. Four out of seven widely distributed species of muried, Rhabdomys pumilio, Saccostomys campestris, Aethomys chrysophilus and Lemniscomys griselda, were shown to be capable of circulating amounts of virus likely to be infective for mosquitoes.


Genetic factors in leprosy: a study of children in Uganda. A group of 20 990 children in Uganda was examined for leprosy over a period of 8 years. There was no evidence that the incidence of leprosy varied according to a child's genetic relationship to a leprosy patients, once allowance had been made for the grade of physical contact.


The behaviour of f2 coliphage in activated sludge treatment. A model activated sludge treatment plant was used which was functionally very similar to a full-scale plant. It was inoculated with f2 coliphage and the titres of virus in the influent, the mixed liquor and the effluent were monitored regularly. The distribution of the virus in the solids and liquid fractions of the mixed liquor was in the ratio of 18:82 and 20.4% of the influent virus was recovered in the effluent. After inoculation was stopped the titre of virus in the solids fraction of the mixed liquor remained high and unaltered for up to 70 h, whereas the value for effluent reverted to the low background titre originally present. These results are discussed in relation to those reported for poliovirus and it is concluded that f2 coliphage is not a suitable model for studies of the behaviour of human enteroviruses.


Immunity to challenge in volunteers vaccinated with an inactivated current or earlier strain of influenza A(H3N2). Volunteers were inoculated with vaccine made from the 30c mutant, A/Port Chalmers/73 or B/Hong Kong/8/73. Preliminary experiments showed that the 30 c strain was antigenically quite close to A/HK/8/68. Volunteers given 30c developed haemagglutination inhibiting antibodies against the 'current' 1973 serotypes (as well as to the vaccine virus) but the titres were less than those after the A/PC/73 vaccine. Volunteers were then challenged with a live attenuated virus, WRL 105, with A/Finland/4/74 antigens, by intranasal inoculation. The rates of infection were 43% after B/Hong Kong/8/73, 20% after 30c and 5% after A/PC/73. This indicated that the 30c gave some protection but that the vaccine prepared from the current strain gave more.


Excess mortality from influenza in a large urban population, Rome, Italy, 1956--76. Influenza activity was studied in the Rome population from 1956 to 1976 by analysis of mortality from respiratory causes and from all causes. During cold weather months, type A influenza virus was associated, as a rule, with epidemic excess deaths at two year intervals while type B virus was prevalent twice during isolation data were also compared with epidemic excess mortality during four consecutive years. The evidence obtained indicated that influenza virus isolation alone does not represent a reliable index of epidemic influenza activity in this population. The proportion of deaths attributed to respiratory causes consistently increased in every epidemic, the most pronounced increases occurring during large epidemics. The break-down by age of deaths from respiratory causes in the course of two epidemic periods showed that the percentage distribution of deaths was essentially the same as in non-epidemic periods. This evidence indicates that the same factors influencing the age-related distribution of mortality from respiratory causes during non-epidemic periods, probably affect the fatal outcome of influenza during epidemics.


Relationship between lymphocyte response to tetanus toxoid and age of lymphocyte donor. An investigation of the in vitro responsiveness of peripheral blood lymphocytes to tetanus toxoid was undertaken to determine the extent to which such reactivity might reflect in vivo reactions to tetanus toxoid immunization. It was found that lymphocyte transformation in the presence of tetanus toxoid, as measured by the uptake of tritiated thymidine, did not differ significantly for the group giving local or systemic reactions to tetanus toxoid and an immune control group. However, a striking inverse relation between lymphocyte reactivity to tetanus toxoid and age of the cell donors of both groups became clear. Such an age-related decline in the in vitro responsiveness of peripheral blood lymphocytes to a specific antigen has apparently had not been reported previously in man.


Two epidemics of diarrhoeal disease possibly caused by Plesiomonas shigelloides. Two epidemics of water-borne diarrhoeal disease involving a total of 1000 persons are reported. In both epidemics, none of the usual bacterial entero-pathogens were recognized and Plesiomonas shigelloides was the only suspect aetiological agent isolated. The ecology of P. shigelloides was investigated in these outbreaks. It was recognized as an inhabitant of fresh surface water and its presence was closely related to warm weather.


The distribution of Vibrio parahaemolyticus in British coastal waters: report of a collaborative study 1975--6. A collaborative survey has shown that V. parahaemolyticus is widely distributed in British coastal waters, sediments and shellfish, especially in southern and western areas. The relatively small numbers in the environment do not present significant health hazards from marine products, provided that processing, storage and distribution are adequate. The presence of this organism in small numbers in British coastal waters or in shellfish should not in itself be regarded as cause for concern.


Epidemiological patterns of Klebsiella colonization and infection in an intensive care ward. Twenty-four per cent of 2315 patients admitted to the intensive care unit of St Thomas's Hospital in the 4 year period from November 1969 became colonized or infected with Klebsiella species. Capsular typing of 986 klebsiella isolates from 551 patients showed that there were 695 patient-isolates, mostly derived from the respiratory tract. Capsular types 47 and 21 were the commonest types and together accounted for 19.9% of the patient-isolates. The 14 commonest types accounted for 47.3% of all patient-isolates and all these strains showed clustering, strongly suggesting a changing common source, cross infection, or both.


Trials of the rodenticide pyriminil against wild house mice (Mus musculus L.). Pen field trials were conducted to assess the performance of the acute rodenticide pyriminil against the house mouse (Mus musculus L.). Four types of poison treatment were carried out using penned family groups of warfarin-resistant mice supplied with alternative plain foods. In each treatment pyriminil was included at 2% in a wholemeal flour/pinhead oatmeal/corn oil bait. Mortality was highest (46/54; 85.2%) when poison bait was offered for 4 days following 3 days of pre-baiting. The same pre-baiting and poisoning technique was adopted in five field trials carried out against mice infesting farm buildings. The efficacy of each poison treatment was estimated from the results of pre- and post-treatment census baitings; treatment success ranged between 53.7% and 96.7%, mean 80.5%. It is concluded that pyriminil treatments are best carried out after a period of pre-baiting and that when pyriminil is used in this manner it is about as effective as zinc phosphide for the control of mice.


Really great returns to medical education? This article reports the value of a medical education as opposed to education for alternative professional careers, using 1970 Census data. When standard net present value computations are performed, it is found that the returns to investment in medical education exceed those to education for alternative professions. When the computations are adjusted for hours worked, physicians' earnings become roughly equal to those of dentists and lawyers but remain clearly superior to those of the other professions considered. Additional information on the earnings of physician cohorts vs. dentist cohorts for the post-Flexner era is also reported.


The impact of occupational licensure of clinical laboratory personnel. This paper develops a model of the economic impact of occupational licensure. The model is then used to estimate the effects of occupational licensure on wages and the division of labor in clinical laboratories and to generate a low-bound estimate of the welfare impact of licensure. Estimates are based on cross-section and time-series data on areas with and without licensure of laboratory personnel. Recent licensure laws have no effect on wages or employment, but older, more stringent laws sharply increase the wages and employment of skilled personnel in laboratories.


The effect of D-penicillamine on mitogen-induced human lymphocyte proliferation: synergistic inhibition by D-penicillamine and copper salts. The effect of D-penicillamine on in vitro mitogen-triggered human peripheral blood lymphocyte DNA synthesis was examined. D-penicillamine caused a modest degree of inhibition of responsiveness. Since D-penicillamine avidly chelates copper, a possible additive effect of D-penicillamine and copper salts on mitogen-induced lymphocyte proliferation was investigated. The addition of various copper salts to D-penicillamine-containing cultures resulted in a marked augmentation in the degree of inhibition observed. Specificity for the synergy between D-penicillamine and copper was suggested by the failure of iron, zinc, or gold salts to augment the inhibition produced by D-penicillamine alone. However, a number of other thiols, but not disulfides, could substitute for D-penicillamine in inhibiting mitogen responsiveness in the presence of copper salts. Preincubation of peripheral blood mononuclear cells with D-penicillamine and CuSO4 resulted in diminished mitogen responsiveness on subsequent culture in the absence of the inhibitors. Data are presented to indicate that preincubation with D-penicillamine and CuSO4 directly affected the ability of the T lymphocytes to respond to mitogens but did not alter the capacity of the monocytes to act as accessory cells in this response.


Interaction between Taenia taeniaeformis and the complement system. Factors present in the cystic bladder fluid of metacestodes of Taenia taeniaeformis and released by these parasites maintained in vitro were shown to interact with the C system in vitro and in vivo. This interaction resulted in the depletion of hemolytic C in normal sera of several species, the generation of anaphylatoxin-like activity in vitro, the conversion of C3, and the production of vascular permeability changes in vivo. The substances appeared to initiate C fixation nonimmunologically via both the alternative and classical pathways. Intravenous administration of parasite factors in rats led to profound depression of circulating C levels and the rats did not become refractory to this effect after repeated dosing over a 3-week period. Circulating levels of hemolytic C were not altered in infected rats over the first 8 weeks of infection. C-dependent inflammatory responses in the skin of rats were inhibited in animals given i.v. doses of parasite factors. The possibility is raised that local consumption of C around the metacestode in vivo could contribute to its successful evasion of inflammation and immune rejection during infection.


Bronchopulmonary macrophage activation in the pathogenesis of hypersensitivity pneumonitis. Repeated intratracheal (IT) inoculation of rabbits with a homogenized, saline suspension of Micropolspora faeni produced bronchopulmonary (BP) histologic lesions resembling those of human hypersensitivity pneumonitis. With an in vitro phagocytic and bactericidal assay, an analysis of BP macrophages from M. faeni-injected rabbits demonstrated activation at both 2 and 4 weeks after the initiation of immunization. No BP macrophage activation was observed in immunized rabbits 6 weeks post-inoculation. BP macrophage activation was capable of recall after 6 weeks in M. faeni-sensitized animals that received a booster IT injection (2 mg) that did not activate "normal" alveolar wash cells. This recall of BP macrophage activation was accompanied by both a marked migration of mononuclear cells into the lung and positive delayed hypersensitivity skin reactions after intradermal injection of M. faeni antigen. Pulmonary histologic examination of sensitized, boosted rabbits suggested an enhanced cellular parenchymal infiltrate when compared with appropriate controls. The above observations confirm the occurrence of immunologically activated BP macrophages in rabbits inoculated witn M. faeni via the respiratory tract route and suggest a correlation between macrophage activation and histopathology.


Immune response to immunization via the anterior chamber of the eye. II. An analysis of F1 lymphocyte-induced immune deviation. Exposure to alloantigen via the anterior chamber of the eye elicits a transient suppression of cellular immunity, whereas humoral immunity is preserved--i.e. F1 LI-ID. The majority of lymphoid cells inoculated into the anterior chamber are retained within the posterior segment of the eye. The latter serves as a depot of alloantigen, allowing the chronic egress of small numbers of cells into the vascular tree. The persistence of this antigen depot is essential to the development of F1 LI-ID. Since there is a preferential distribution of cells that migrate from the eye to the spleen, the functional integrity of the latter is also necessary to elicit F1 LI-ID. It is concluded that an anatomically intact spleen, i.v. presentation of antigen, and persistence of antigen within the eye are all important to the elicitation of this phenomenon.


Isolation of an Fcgamma-binding protein from the cell membrane of a macrophage-like cell line (P388D1) after detergent solubilization. Lactoperoxidase-catalyzed iodination, NP-40 lysis, and subsequent affinity chromatography on IgG-Sepharose were used in an attempt to define some of the molecular properties of the Fc receptor of P388D1, a macrophage-like mouse tumor line. Radioiodinated material retained on columns of Sepharose coupled either to monomeric mouse IgG2a or monomeric human IgG1 appeared on SDS polyacrylamide gel electrophoresis to contain principally three labeled components, a major band of about 57,000 m.w. and two minor bands of 28,000 and 24,000 m.w. The mobilities of these components changed little on reduction, which suggested that they represented single polypeptide chains, An identical pattern was obtained with Sepharose-linked Fc fragments of human IgG1, but neither Fab fragments of IgG1 nor IgM appeared to bind these components. Since the specificity of binding to the immobilized proteins is the same as that observed in vivo, it is postulated that these proteins represent either all or some portion of the P388D1 Fc receptor.


Enumeration and ultrastructure of C4-producing free alveolar cells from guinea pig lung. Free alveolar cells from guinea pig lung producing the fourth emoponent of C (C4) were identified, enumerated, and characterized by using anti-C4 Fab-peroxidase conjugates in conjunction with transmission electron microscopy. The C4-producing cell population consisted of: 1) alveolar macrophages (AM); 2) less well differentiated phagocytes similar in morphology to exudate macrophages; and 3) weakly phagocytic secretory cells with numerous profiles of rough-surfaced endoplasmic reticulum (ER). Internal immunolabeling allowed the visualization of C4 in the ER, perinuclear space, and Golgi complex of producer cells and its release at cell surfaces; synthesis of C4 in vitro was sensitive to inhibitors both of protein synthesis and messenger RNA function. The percentage of free alveolar cells from normal animals competent for C4 production as indicated by cell surface immunolabeling was approximately 1% of the total cells obtained by lavage. Transnasal infection with Listeria monocytogenes, generation of a pulmonary granulomatous reaction by i.v. injection of heat-killed BCG, and aerosol infection of nonvaccinated animals with Myco-bacterium tuberculsois each resulted in an increase in numbers of AM and exudate macrophage-like free alveolar cells competent for C4-production.


Immune response in guinea pigs to two different lipid conjugates of bovine serum albumin. A lipid conjugate of BSA was made by coupling dodecylamine to the COOH groups of BSA (DA-BSA). Studies on the immunogenicity of this material and one made by coupling dodecanoic anhydride to NH2 groups (D-BSA) demonstrated that cell-mediated immunity could preferentially be generated by lipid conjugation to an antigen as measured by delayed skin reaction, invitro blast transformation, and antibody formation. DA-BSA was found superior by both in vivo and in vitro tests. Both the conjugates retained the ability to suppress delayed type hypersensitivity, but only DA-BSA retained the ability to elicit optimum skin reaction in a sensitized animal and to precipitate anti-BSA antibody.


Anisakis and eosinophil. I. Detection of a soluble factor selectively chemotactic for eosinophils in the extract from Anisakis larvae. Studies were undertaken in order to determine if Anisakis larva itself has the ability to attract eosinophils. Extracts from Anisakis larvae were examined for both neutrophil and eosinophil chemotactic activities with both in vivo and in vitro assay systems. When the soluble extract was injected intradermally into normal guinea pigs, a profound accumulation of eosinophils was observed at the injection site. The cells started to accumulate at the site within 1 hr and the number of eosinophils at the site reached its peak at 8 hr after the injection of the extract. Such eosinophilic accumulation was enhanced in a dose-response fashion over the range of 0.5 to 50 micrograms protein. The strong chemotatic activity of the Anisakis extract for eosinophils was confirmed when in vitro chemotaxis assays were performed with Boyden chemotatic chambers. Interestingly, no chemotactic activity for neutrophils was found at those concentrations of the extract with which the eosinophil effect was observed. These results indicate that the factor described here, in addition to the various known immunologic factors, may play an important role in the development of eosinophilia in anisakiasis.


Plasminogen activator of guinea pig basophilic leukocytes: probable localization to the plasma membrane. The plasminogen activator (PA) activity of guinea pig basophil-enriched leukocyte preparations was localized to basophils, and not to contaminating lymphocytes and eosinophils, by correlating PA activity with basophil frequency and, more directly, by means of an improved cytochemical method here described. PA activity was fully expressed in living cells in the absence of immunologic stimuli and was suppressed/lost to a variable extent by different techniques of cell disruption. Conversely, killed, but not living, basophils expressed significant plasminogen-independent fibrinolytic activity, presumably reflecting access of cytoplasmic proteases of broken basophils to fibrin substrate. The PA activity of intact cells was destroyed by gentle trypsinization under conditions that did not impair cell viability. When disrupted cells were ultracentrifuged on a sucrose density gradient, PA activity was absent from purified granules and was confined to fractions containing cell membranes. The simplest explanation of these data is that guinea pig basophils have PA activity associated with their plasma membranes. This conclusion has several important implications for basophil functions in cell-mediated and other immunologic reactions in vivo.


Natural killing in immunodeficient patients. Natural killing (NK) capacity was evaluated in peripheral blood mononuclear cells from 14 patients with well defined primary immunodeficiency disorders and compared with the activity of those cells in antibody-dependent cell-mediated cytotoxicity (ADCC) assays against antibody-coated erythrocyte (killed primarily by monocytes) and lymphoid or tumor targets (killed exclusively by lymphoid cells). A selective inability to lyse antibody-coated lymphocyte targets was observed with cells from patients with x-linked agammaglobulinemia, suggesting the involvement of either a different lymphocyte subpopulation or membrane receptor for NK and ADCC, or that a different functional susceptibility exists for the two types of killing. The only immunodeficiency state in which lymphocyte NK activity was found to be lacking was severe combined immunodediciency disease.


Membrane receptors of mouse leukocytes. I. Two types of complement receptors for different regions of C3. Mouse leukocytes were studied for membrane receptors for the third C component by rosette formation with C coated erythrocytes (EAC). Methods were devised for the preparation of EAC complexes containing either mouse C3b or mouse C3d. EAC 1-3dmo were prepared from EA treated with whole mouse serum while EAC 1-3bmo were produced from EAC 142hu treated with whole mouse serum containing sodium suramin. The specificity of the EAC complexes for mouse leukocytes was confirmed by inhibition experiments using fluid phase human C3d. Low concentrations of fluid phase human C3d inhibited EAC1-3dmo rosettes but failed to inhibit EAC 1-3bmo rosettes. Eight-fold higher concentrations of fluid phase C3d caused partial inhibition of EAC1-3bmo rosette formation with lymphocytes, but not with other types of murine leukocytes. Thus mouse leukocytes apparently contain the same two types of C receptors as do human and guinea pig leukocytes. Mouse CR1 is specific for a non-C3d region of C3b, (possibly analogous to human C3c) whereas mouse CR2 is specific for both C3d and the C3d region of C3b.


Stimulation of the synthesis and release of lipids in tumor cells under attack by antibody and C. Antibody-sensitized line-1 or line-10 tumor cells treated with GPC (TAC) incorporated fatty acids into complex cellular lipids and released increased amounts of fatty acids within 5 to 10 min after the addition of GPC as compared to control cells. This effect was dependent on the concentration of GPC used; however, under conditions where the cells were not killed, the enhanced synthesis and release of lipids were not dependent on the antibody concentration used to sensitize the cells. Treatment of the cells with antibody alone, GPC alone, or antibody plus heat-inactivated GPC did not result in enhanced synthesis or release of lipids. No enhancement in DNA, RNA, or protein synthesis in TAC was noted. Line-1 cells, which can be killed by GPC when sensitized with excess anti-Forssman IgM antibody, demonstrated enhanced lipid synthesis within 1 to 3 min after the addition of GPC to the antibody-sensitized cells, before measurable killing of the cells had occurred. This effect persisted in the surviving cells when tested 5 and 10 min after the formation of TAC. Addition of GPC deficient in C4 to antibody-sensitized cells did not result in enhanced lipid synthesis or release. These data suggest that the synthesis of macromolecules of which lipids are a major component is of central importance for the ability of the cells to resist antibody-GPC mediated attack.


Aging and the immune response. I. Antibody formation and chronic infection in Toxoplasma gondii-infected mice. Studies were performed to determine the effect of aging on the antibody response and cyst formations after infection with a relatively avirulent strain of the intracellular parasite Toxoplasma gondii. When compared with young mice (4 months), aged mice showed a significant decrease in the magnitude of humoral immune response to infection. This decrease was observed at the peak of the acute infection and also during chronic infection. Evaluation of the presence of Toxoplasma cysts, a measure of the latent infection, revealed that the numbers of tissue cysts present 11 weeks after infection increased with the age of the mice at time of infection. The larger numbers of cysts in older mice which had received the same inoculum size of T. gondii as young mice, together with our previous observations of increased susceptibility of these older mice to T. gondii, suggest that an age-related decrease in the early immune response to this infection allows an increased multiplication of the organism in vivo, leading to increased cyst numbers or death.


DNA synthesis and proliferation of human lymphocytes in vitro. I. Cell kinetics of response to phytohemagglutinin. The response of human lymphocytes to phytohemagglutinin (PHA) stimulation was assessed in detailed kinetic studies, in order to define conditions permitting and regulating proliferation. By utilizing live and dead cell counts, cell cycle characterizations, and calculations of rates of entry into S phase and mitosis, it was demonstrated that dilute culture conditions (2 X 10(5) cells/ml) enable lymphocytes to proliferate for a period of 5 or 6 days. Although cell division occurs in concentrated cultures (2 X 10(6) cells/ml), net proliferation is not demonstrable due to extensive cell death and earlier decay in the proliferative response. It is projected that without cell death the cell count would rise more than 6-fold in a dilute culture and 2-fold in a concentrated culture under these growth conditions. The kinetic analyses did not demonstrate separate PHA dose thresholds for blastogenesis and entry into S phase. High doses of PHA stimulated a suboptimal response by causing excessive cell death without reducing initial entry into the cell cycle. These results provide a kinetic explanation for the frequent observation of little net proliferation despite substantial 3H-thymidine incorporation in PHA-stimulated lymphocyte cultures. Cell death is identified as a parameter that must be considered in the interpretation of in vitro lymphocyte stimulation studies, and methods for quantifying the role of cell death are demonstrated.


DNA synthesis and proliferation of human lymphocytes in vitro. II. Characterization of the DNA newly synthesized after phytohemagglutinin stimulation. DNA newly synthesized by phytohemagglutinin-(PHA) stimulated human lymphocytes has been analyzed for the possibility that all DNA synthesis may not represent premitotic genome duplication. Equilibrium density gradient characterization of bromodeoxyuridine-(BUdR) substituted DNA demonstrates semi-conservative DNA replication, without evidence for repair synthesis. Experiments to detect selective replication (amplification) of a portion of the DNA involved measurement of reassociation kinetics as well as measurement of the kinetics of BUdR appearance in prelabeled DNA. The presence of large quantities of amplified DNA has been excluded, although amplification cannot be ruled out completely by these techniques. Finally, the kinetics of DNA release from dead lymphocytes is characterized, and factors tending to reduce the complexity of released, labeled DNA are identified.


Opsonic activity of guinea pig serum is reduced by in vivo administration of carcinogenic nitrosamines. The administration of each of four carcinogenic nitrosamines to normal guinea pigs resulted in decreased opsonic activity of their sera but did not affect the capacity of their peritoneal macrophages to phagocytize particles opsonized with normal serum. Diphenylnitrosamine, a noncarcinogenic analogue, had no significant effect on opsonic or phagocytic activity.


A simplified procedure for cyclic nucleotide radioimmunoassay and its application to human blood leukocytes. Rapid treatment of leukocyte suspensions was found to be an effective alternative to acid treatment in the preparation of these cells for radioimmunoassay of cyclic GMP and cyclic AMP. A 2-sec heating to boiling temperature, followed by sonication and micropore filtration, was employed. This procedure adequately inactivated or removed enzymes and binding proteins that can alter cyclic nucleotide concentrations or otherwise interfere with the radioimmunoassay. Moreover, heating in this manner did not appear to affect the stability of cyclic nucleotides or cause significant formation of cyclic GMP from endogenous GTP. Recovery of cyclic nucleotides after heating and filtration was high (89-96%), making possible the measurement of both cyclic GMP and cyclic AMP in small cellular samples. Variation in cyclic nucleotide recovery was small (+/- 2-4% SD), therefore individual recovery determinations were unnecessary.


Systemic protection by antioxidants against UVL-induced erythema. An antioxidant supplemented diet provided marked systemic protection against ultraviolet light mediated erythema in hairless mice. Among the individual constituents of the diet, butylated hydroxytoluene was most effective whereas glutathione and vitamins C and E afforded negligible protection. The mixture of antioxidants, and butylated hydroxytoluene individually, demonstrated diminished, but significant, protection when applied topically. The safety of this systemic photoprotectant and its clinical relevance at present is unknown.


The effect of aluminium chlorhydrate on sweat gland activity in cattle. Topical application of aluminium chlorhydrate had no appreciable antiperspirant action on the epitrichial glands of cattle. There was no evidence of penetration of the salt into the dermis or of any change in the morphology of the glands. It is probable that the antiperspirant activity of aluminium chlorhydrate in the human axilla, is only on the atrichial glands.


Route of contact sensitization and in vitro lymphocyte transformation. A guinea pig skin extract conjugate with dinitrofluorobenzene elicited significant in vitro transformation of cultured lymphnode lymphocytes from 19 of 27 guinea pigs sensitized by footpad injection of dinitrochlorobenzene in Freund's complete adjuvant, as compared to only 1 of 26 guinea pigs topically sensitized to dinitrochlorobenzene. Topically sensitized guinea pigs appear to be more appropriate models for contact allergy in man than guinea pigs sensitized by other methods. Other sensitization procedures are likely to produce more heterogeneous forms of sensitization, with features of contact allergy, tuberculin-type allergy, antibody-mediated hypersensitivity and cutaneous basophile hypersensitivity.


Mitotic activity of epidermal melanocytes in UV-irradiated mouse skin. The role of mitosis in the increase in the epidermal melanocyte population following repeated ultraviolet (UV) irradiation has been studied in C57B1 mice. Cumulative labeling with 3H-methyl-thymidine (3HTdr) was used in combination with autoradiography. A 4-6 fold increase in the melanocyte population was associated with an isotope labeling of 65-80% of the melanocytes. This indicates that cell division during the irradiation period can account for the total increase in the number of melanocytes. Labeled melanocytes were also found in the epidermis from unirradiated skin. Thus, the epidermal melanocytes seem to constitute a dynamic cell population under continuous renewal.


Identification of topical photosensitizing agents in humans. A method is described for the detection of topical photosensitizers in humans. Test agents were applied to the untanned midback under an occlusive dressing for 6 hr and then exposed to broad-spectrum radiation containing UV-A and visible wavelengths from a Xenon arc source. Well-known topical photosensitizers were readily identified. It was found that with certain poorly-penetrating substances, such as water-soluble dyes, applications to scarified skin were necessary to reveal phototoxic activity. In addition, these dyes were activated by wavelengths in the visible region of the spectrum.


A method to evaluate skin moisturizers in vivo. Blank demonstrated that water is the only plasticizer of human stratum corneum. Moisturizers attempt to add and hold water within the horny layer, and have previously been evaluated subjectively or in vitro. By passing a stream of dry nitrogen over skin, then through an electrolytic moisture analyzer, moisture present at the skin surface will be detected. This technique has been utilized to measure transepidermal water loss and the inhibition of water loss from skin by various investigators. However, when this same procedure is done on normal human skin, higher values indicate the presence of more moisture. The more moisture detectable at the surface, the more moisture available for keratin to absorb. By applying the moisturizers to plastic film and the skin, it can be demonstrated that water is not detectable after a few minutes on the plastic, but is detectable on skin at higher than control (transepidermal water loss) values for several hours. The technique can demonstrate enhanced moisturization from plastic occlusion and wet dressings as well as the drying effect of benzoyl peroxide gels. It allows objective rank ordering of moisturizers by monitoring moisture enhancement rather than occlusivity of applied substances.


Distensibility of the vascular bed in subcutaneous tissue in generalized scleroderma. Passive distensibility of resistance vessels in subcutaneous tissue of hand and leg was examined in 6 normals and 8 patients suffering from generalized scleroderma of the acrosclerosis type. Local paralysis of smooth muscle was obtained by injection of papaverine and increase in transmural pressure by lowering the limb. Blood flow was estimated by the local 133Xenon wash-out method. Distensibility of blood vessels in subcutaneous tissue in generalized scleroderma was not decreased to a significant degree, whereas, both in patients and normals, the blood vessels of the leg exhibited diminished distensibility compared to blood vessels of the hand. The findings suggest that the morphologic changes of vascular and extravascular tissue observed in sclerodermic skin do not affect distensibility of the vascular bed in subcutaneous tissue.


In vitro activity of cephalosporins against methicillin-resistant, coagulase-negative staphylococci. The in vitro activity of five cephalosporins, gentamicin, and vancomycin was determined against 41 clinical isolates of methicillin-resistant, coagulase-negative staphylococci. Results obtained with disk diffusion and agar dilution methods failed to show complete cross-resistance between methicillin and four of the five cephalosporins despite the use of a high-salt medium and a large inoculum. Thirty-six (88%) of the 41 isolates were sensitive to cephalothin by a standardized disk diffusion method, whereas 23 isolates (56%) were sensitive to cephalothin with use of an agar dilution method and a large inoculum. Of these 23 isolates, only 11 (47%) were both inhibited and killed by less than or equal to 6.25 microgram of cephalothin/ml. The inhibitory and bactericidal activity of gentamicin was greater than that of vancomycin and cephalothin. These results suggest that the disk diffusion technique has limitations in determining the in vitro activity of cephalosporins against methicillin-resistant, coagulase-negative staphylococci and that cephalothin exhibits poor bactericidal activity against these same isolates.


Epidemiology of hepatitis B surface antigen (HBsAg) and antibody to HBsAg in hospital personnel. The epidemiology of hepatitis B in hospital personnel was studied by testing of sera from 3,770 employees of the Medical School of Hannover (Hannover, West Germany) for hepatitis B surface antigen (HBsAg) and its corresponding antibody (anti-HBs) by solid-phase radioimmunoassay. An average prevalence of 2.2% for HBsAg and 11.7% for anti-HBs was found. Physicians (18.2%), nurses (20.1%), and members of the cleaning service (26.3%) showed the highest frequencies of HBsAg or anti-HBs carriage. In a study of age- and sex-matched personnel, nurses showed a significantly (P less than 0.01) higher rate of infection than a control group with less exposure to infectious materials. The frequency of HBsAg or anti-HBs was highest in persons associated with dialysis (31.3%), anesthesiology (31.0%), ophthalmology (29.4%, neurosurgery (28.0%), and surgery (24.4%). The rate of infection was significantly higher in surgical departments (24.4%) than in nonsurgical ones (13.3%). Persons who had been nursing patients with hepatitis were significantly (P less than 0.05) more frequently carriers of HBsAg or anti-HBs than a comparable control group.


Serologic studies of human genital mycoplasmas: distribution of titers of mycoplasmacidal antibody to Ureaplasma urealyticum and Mycoplasma hominis in pregnant women. Titers of mycoplasmacidal antibody to the human genital mycoplasmas Ureaplasma urealyticum and Mycoplasma hominis were determined using genital isolates from pregnant patients as antigens and comparing these isolates with the 11 prototypic reference strains for U. urealyticum and the seven reference strains for M. hominis. Virtually all titers that were detected with use of the patient's own isolates were detected by the 11 reference strains of U. urealyticum and by the seven reference strains of M. hominis. Serologic surveys of pregnant women who harbored either or both mycoplasmas in vaginal cultures indicated that antibody to M. hominis was found more commonly than antibody to U. urealyticum. It was demonstrated that significant postpartum rises in titers of antibody to M. hominis were correlated with the presence of these mycoplasmas in genital cultures. Postpartum rises in titer of antibody were particularly likely to occur in women with low titers of mycoplasmacidal antibody in serum at the time of delivery. Approximately 88% of the women who were colonized with M. hominis showed significant changes in titer of antibody to M. hominis throughout an apparently normal pregnancy; only 40% of the women who were colonized with U. urealyticum showed such changes in titers to U. urealyticum. Statistical analysis showed that mean log titers of antibody to both mycoplasmas at the first prenatal visit were significantly associated with the number of pregnancies experienced by these women.


In vitro susceptiiblity of Mycobacterium fortuitum and Mycobacterium chelonei to amikacin. Infections due to Runyon group IV atypical mycobacteria, Mycobacterium fortuitum and Mycobacterium chelonei, in humans have been difficult to treat in the past because of the organisms' resistance to all of the conventional antimycobacterial drugs. Determinations of minimal inhibitory concentrations (MICs) by the agar dilution method suggest that amikacin may be useful in the treatment of infections due to M. fortuitum and M. chelonei. Further support for the efficacy of antimicrobial agents with good in vitro activity in the treatment of infections with M. fortuitum and M. chelonei will depend on correlation of future clinical experience with standardized data on the MICs for these organisms.


Durability of antibody titers induced by RA27/3 rubella virus vaccine. The RA27/3 rubella virus vaccine induced titers of hemagglutination-inhibiting antibody that were slightly lower than those induced naturally, but after an initial decline the titers remained stable even in the absence of reexposure to rubella virus.


Surgical microscopy of the nose. New frontiers in nasal diagnosis and treatment. The need for microscopic rhinoscopy is obvious to otolaryngologists who have used the operating microscope in otology and laryngology. It provides excellent exposure, brilliant illumination, magnification and the opportunity for bimanual instrumentation, teaching and documentation. The development of the procedure is discussed and the technique of using the method in endonasal diagnosis and surgery is illustrated.


Carcinomas of the nasal septum.

Carcinomas of the nasal septum. Fourteen new cases of primary squamous cell carcinoma of the nasal septum are presented. Their inclusion brings the total reported in the literature to 97. Early, confined neoplasms are amenable to cure by surgical excision, external irradiation or radium implants. Prognosis does not correlate with histological grade of the neoplasm but is inversely related to the size of the carcinoma at the time of diagnosis and the presence of metastases.


Spontaneous haematoma in the head and neck. Three cases of spontaneous haematoma in the head and neck are presented. Whereas one case was associated with lingual haemangiomatosis, no source of bleeding could be demonstrated in either a chronic, spontaneous orbital blood-cyst or an acute retropharyngeal haematoma.


Plasmacytoma of the larynx.

Plasmacytoma of the larynx. Plasmacytoma of the larynx is rare; thirty-one cases are reported in the literature and in this paper a further case is presented. The difficulty in diagnosing plasmacytoma is mentioned. Investigations to exclude multiple myeloma are mandatory to be sure that the disease is a localized lesion. Treatment of a localized lesion is essentially by radiotherapy but in the case of widespread or systemic disease, chemotherapy is recommended. A scoring system utilizing different parameters is suggested to decide whether the case has passed the borderline between local and systemic disease, a line which is invariably difficult to define.


Hearing loss in carcinomatous meningitis. Although the syndrome of carcinomatous meningitis presenting as multiple cranial neuropathies is well known, hearing loss as a presenting complaint has not been previously stressed. Five cases of carcinomatous meningitis beginning as hearing loss are presented and the clinical and laboratory features discussed. A thorough review of the literature summarizes the pathology, pathogenesis and recommended modes of therapy of this disease and the importance of early diagnosis and treatment is stressed.


Dominant hereditary nerve deafness.

Dominant hereditary nerve deafness. A family of seventy-six members showing a dominant progressive nerve deafness is reported and compared with previously reported cases. It is suggested that the basic site of the lesion may be in the vascular system.


Salivary gland choristoma of the middle ear. A case is described in which a middle-ear salivary gland choristoma was found in association with dehiscence of the horizontal part of the facial nerve and abnormality of the stapes and incus. It is suggested that these features may constitute an abnormal development syndrome.


Embryonal rhabdosarcoma of the middle ear and mastoid. Two cases of embryonal rhabdomyosarcoma, arising from the middle ear, are described. It is a rare mesenchymal tumour of childhood, with a very poor prognosis. The literature is briefly reviewed and the difficulties encountered in diagnosis are outlined.


A quantitative analysis of fine structure and drug metabolism in livers of clofibrate-treated young adult and retired breeder rats. The effects of clofibrate on the fine structure and drug-metabolizing capacity of livers of normolipidemic young adult virgin (YA) and hypercholesterolemic retired breeder (RB) male rats were measured by morphometric and biochemical procedures. The oral administration of clofibrate for 7 days significantly increased liver weight and reduced the cholesterol concentrations in the serum and liver tissue in both groups of animals. The hepatic triglyceride (TG) concentration and the volume of cytoplasmic lipid droplets, presumably TG, as well as the serum TG concentration, increased only in the drug-treated RB rats. Clofibrate treatment resulted in significant increases in the volumes of the hepatocytes and their constituent mitochondria and microbodies and caused a proliferation of the smooth-surfaced endoplasmic reticulum. Although the magnitude of the hypocholesterolemic response was considerably greater in the RB animals, the morphological changes were much more marked in the YA group. However, the surface area of the rough-surfaced endoplasmic reticulum was reduced in the livers of the drug-treated RB rats. NADPH cytochrome c reductase specific activity was significantly increased in both the RB and YA animals, but the concentration of cytochrome P-450 (per mg microsomal protein) increased only in the YA rats. Neither the cytochrome b5 concentration nor the rate of ethylmorphine N-demethylation was significantly affected by clofibrate administration. The results suggest that there is no positive correlation between the hypocholesterolemic response to clofibrate and the degree of subcellular changes in the hepatocytes and that this hypolipidemic drug elicits a minimal effect on the concentrations of the components of the hepatic microsomal drug-metabolizing system.


Analysis of fatty acid methyl esters by a gas--liquid chromatography--chemical ionization mass spectrometry computer system. The technique of gas-liquid chromatography-chemical ionization mass spectrometry can easily identify trace peaks and unresolved peaks on gas-liquid chromatography, utilizing MH(+) ions of chemical ionization mass spectra. In polyunsaturated fatty acid methyl esters such as C22:5 and C22:6, the determination of molecular weights that are difficult to determine by electron impact mass spectrometry could be easily identified by chemical ionization mass spectrometry. The identification could be performed even more easily from a mass chromatogram obtained by means of a gas-liquid chromatography-chemical ionization mass spectrometry-computer system. This technique was applied to analysis of fatty acid methyl esters of triglycerides, cholesteryl esters, nonesterified fatty acids, and phospholipids obtained from normal human serum. It was observed that odd-carbon-number fatty acids contained more isomers of different types than even-carbon-number fatty acids, whether the fatty acids were saturated or unsaturated. As for the types of isomers, we presume the existence of iso, anteiso, branched, cyclic fatty acids, and, in the case of unsaturated fatty acids, positional isomers. The qualitative determination of the group of fatty acid isomers containing 20 carbon atoms was also investigated as well as the application of procedures for quantitative determination. Sensitivities for saturated fatty acids differ from those for unsaturated fatty acids of the same carbon number, and sensitivities for unsaturated fatty acids of the same carbon number differ with the number of double bonds because the amounts of MH(+) - 32 and MH(+) - 32 - 18 fragment ions formed are different.


Determination of double bond positions of unsaturated fatty acids by a chemical ionization mass spectrometry computer system. After stereospecific oxidation, trimethylsilylated methyl esters of mono- and diunsaturated fatty acids were analyzed by combined gas-liquid chromatography-chemical ionization mass spectrometry. The positions of original double bonds were deduced from the fragment ions produced by the cleavage of the carbon-carbon bond between two trimethylsilyl ethers. These fragment ions were recorded at m/e 187 and 259 in the case of 16:1(n-7), at m/e 187 and 287 in the case of 18:1(n-7), at m/e 215 and 259 in the case of 18:1(n-9), and at m/e 172 and 259 in the case of 18:2(n-6), respectively. The diastereoisomers of monounsaturated fatty acids can be discriminated by comparing the intensities of the fragment ions at m/e 253 and 285 in the case of 16:1 and at m/e 281 and 313 in the case of 18:1. The diastereosiomers of diunsaturated fatty acids may also be distinguished from each other by comparing the intensities of the fragment ions formed by the loss of trimethylsilyl function from the characteristic ions. Mono- and diunsaturated fatty acids in liver may be dominantly in cis- 18:1(n-1) and cis-cis-18:2(n-6) as determined by a mass chromatographic technique.


Occurrence of sulfated 5alpha-cholanoates in rat bile. Bile acids in bile from male and female rats with cannulated bile ducts have been analyzed by repetitive scanning gas-liquid chromatography-mass spectrometry after initial fractionation of conjugate classes on diethylaminohydroxypropyl Sephadex LH-20. Sex differences were observed in the amounts and types of bile acids in the sulfate fraction. The proportion of total bile acids excreted as sulfates was higher in female (0.9-1.3%) than in male (0.1-0.2%) rats. Most of the sulfated bile acids had a 5alpha configuration, allochenodeoxycholic acid being the major compound in bile from female rats. This bile acid was also present in the nonsulfate fraction but could not be found in bile from male rats. The results indicate that gas-liquid chromatography-mass spectrometry has to be used to provide sufficient specificity in the bile acid analyses. Thus, compounds from the sulfate fraction having the retention times of cholic and chenodeoxycholic acid derivatives were found to be due to derivatives of the 3beta,5alpha-isomers of these bile acids.


Absolute configuration of pentahydroxy bile alcohols excreted by patients with cerebrotendinous xanthomatosis: a circular dichroism study. The absolute configurations of the C27 pentahydroxy bile alcohols present in bile and feces of two patients with cerebrotendinous xanthomatosis (CTX) were determined by circular dichroism (CD) spectroscopy. The CD spectra of 5beta-cholestane-3alpha,7alpha,12alpha,24alpha,25-pentol in the presence of Eu(fod)3 [tris(1,1,1,2,2,3,3-heptafluoro-7,7-dimethyloctane-4,6-dionato) europium (III)] exhibited a negative Cotton effect and was assigned to 24R absolute configuration. Conversely, 5beta-cholestane-3alpha,7alpha,12alpha,24beta,25-pentol showed a strong positive Cotton effect and was assigned the 24S configuration. These assignments were based upon comparison with a model compound, 5-cholestene-3beta,24(R),25-triol, whose single-crystal X-ray structure has been determined. The importance of these data is to establish a structural mechanism for the conversion of 5beta-cholestane-3alpha,7alpha,12alpha,24S,25-pentol rather than 5beta-cholestane-3alpha,7alpha,12alpha,24R,25-pentol into cholic acid in man as well as in animals.


Preparation of 24(R)- and 24(S)-5beta-cholestane-3alpha,7alpha,24-triols and 25(R)- and 25(S)-5beta-cholestane-3alpha,7alpha,26-triols by a hydroboration procedure. This report describes a new and convenient method for the preparation of 5beta-cholestane-3alpha,7alpha,24-triol (24R and 24S) and 5beta-cholestane-3alpha,7alpha,26-triol (25R and 25S) starting from 5beta-cholestane-3alpha,7alpha,25-triol. Dehydration of the latter with acetic anhydride and glacial acetic acid yielded a mixture of 5beta-cholest-24ene-3alpha,7alpha-diol and the corresponding delta25 compound. Hydroboration and oxidation of the delta24 unsaturated bile alcohol resulted in the formation of 5beta-cholestane-3alpha,7alpha,24-triol. 5beta-Cholestane-3alpha,7alpha,26-triol and 5beta-cholestane-3alpha,7alpha-diol were obtained from the delta25 bile alcohol. In each case the bile alcohols epimeric at C-24 and C-25 were resolved by analytical and preparative thin-layer chromatography and characterized by gas-liquid chromatography, infrared-, proton magnetic resonance-, and mass spectrometry. Tentative assignment of the 24R, 24S and 25R, 25S configurations was made on the basis of molecular rotation differences. These epimeric bile alcohols will be useful for biological studies of chenodeoxycholic acid biosynthesis.


Determinants of fatty acid and alcohol monomer activities in mixed micellar solutions. The determinants of monomer activities of lipids dissolved in micellar bile salt solutions have been studied using polyethylene discs as the organic phase of a partitioning system. The studies show that fatty acids and alcohols interact with micelles as a partitioning system so that the monomer activity is determined by micelle volume and the lipid's partition coefficient as well as mass of lipid in the solution. Influence of the partition coefficient is seen in the dependence of monomer activity on chain length, unsaturation and carboxyl or alcohol polar groups. Dependence on chain length is equivalent to an incremental free energy of approximately -700 cal. mol(-1) per methylene group. Substitution of an alcohol group for the carboxyl group at pH 7.4 decreases monomer activity by a factor of 900. Expansion of taurodeoxycholate micelles with 5mM monooleoylglycerol slightly decreases monomer activity whereas solutions of lipids in taurocholate have relatively greater monomer activities, demonstrating the influence of volume of the micelle organic phase. With constant micelle structure, monomer activity was linearly dependent on lipid mass in the system as predicted by partitioning theory. Addition of low concentration of lecithin, lysolecithin, or monoacylglycerol to the solutions had only small effects on the monomer activities consistent with the small change in total micelle organic phase. Data provided allow calculation of monomer activities of fatty acids and alcohols in many complex micellar solutions. Such data are important for evaluating such processes as intestinal absorption and gallstone formation and dissolution.


Effect of biliary obstruction on 26-hydroxylation of C27-steroids in bile acid synthesis. The effect of biliary obstruction on side chain hydroxylations in the biosynthesis and metabolism of bile acids was studied in the rat. For comparison, several other hydroxylation reactions in bile acid biosynthesis and metabolism were assayed. Biliary obstruction inhibited microsomal 26-hydroxylation of 5beta-cholestane-3alpha,7alpha-diol and microsomal 25- and 26-hydroxylation of 5beta-cholestane-3alpha,7alpha-12alpha-triol. Microsomal 7alpha-hydroxylation of cholesterol and 6beta-hydroxylation of lithocholic acid acid increased significantly, whereas the increase in microsomal 12alpha-hydroxylation of 5beta-cholestane-3alpha,7alpha-diol was less. Mitochondrial 26-hydroxylation of cholesterol, 5-cholestene-3beta,7alpha-diol, and 7alpha-hydroxy-4-cholesten-3-one was stimulated, whereas 26-hydroxylation of 5beta-cholestane-3alpha,7alpha-diol was not affected and that of 5beta-cholestane-3alpha,7alpha,12alpha-triol was markedly inhibited. The results indicate that mitochondrial 26-hydroxylation, particularly of substrates that primarily are precursors of chenodeoxycholic acid, plays a more important role in bile acid biosynthesis under conditions of biliary obstruction than under normal conditions.


Ruminal hydrogenation of cholesterol.

Ruminal hydrogenation of cholesterol. Cholesterol was hydrogenated by anaerobic incubation with sheep rumen fluid for periods up to 20 hr. The principal product of cholesterol hydrogenation was identified as coprostanol. Cholesterol could be protected against in vitro ruminal hydrogenation by encapsulation in a matrix of formaldehyde-treated casein. Formaldehyde-treated casein-cholesterol preparations were also shown to be protected against hydrogenation in vivo and, when supplements containing 1 g per day of protected or unprotected cholesterol were fed to sheep over a period of 8-9 weeks, there were marked differences in the plasma cholesterol response. The plasma cholesterol of the sheep fed protected cholesterol increased by at least 60%. The plasma cholesterol of the sheep fed unprotected cholesterol also tended to increase during the first 5 weeks of supplementation but thereafter declined to almost control levels at 8 weeks.


Total synthesis of stereospecific sphingosine and ceramide. A small-scale synthesis of the four sphingosine stereoisomers (d-erythro, l-erythro, d-threo, and l-threo) and lignoceroyl d- and l-erythro-sphingosines, which is suitable for synthesis of tritium-labeled compounds, is described. Ethyl dl-erythro-2-acetamino-3-hydroxy-4t-octadecenoate was esterified with l(+)-acetylmandeloyl chloride and the two diastereomers obtained were separated from each other by thin-layer or column chromatography. Each diastereomer was subjected to ethanolysis to obtain ethyl d- or l-erythro-2-amino-3-hydroxy-4t-octadecenoate which was then reduced with LiAlH(4) or NaBH(4) to yield d- or l-erythro-sphingosine. d-erythro-[1-(3)H]Sphingosine with high specific activity was prepared by using LiAl(3)H(4) in the last step. d- and l-threo-sphingosines were synthesized from ethyl dl-threo-2-acetamino-3-hydroxy-4t-octadecenoate by using a similar procedure. Ceramide (lignoceroyl sphingosine) was prepared either by acylating sphingosine or by the following new method. Ethyl dl-erythro-2-amino-3-hydroxy-4t-octadecenoate was converted to the N-lignoceroyl derivative and esterified with l(+)-acetylmandeloyl chloride. The two diastereomers obtained were separated and each isomer was treated with a catalytic amount of sodium ethoxide. One of the products, ethyl d-erythro-2-lignoceroylamino-3-hydroxy-4t- octadecenoate, was reduced with NaBH(4) to yield ceramide. N-palmitoyl dl-erythro-sphingosine was also prepared using an identical procedure. N-lignoceroyl d-erythro-[1-(3)H]sphingosine was prepared by NaB(3)H(4) reduction of the corresponding amide ester. A doubly labeled ceramide, [1-(14)C]lignoceroyl [1-(3)H]sphingosine, containing high specific activity, was prepared by mixing the above N-lignoceroyl d-erythro-[1-(3)H]sphingosine and N-[1-(14)C]lignoceroyl d-erythro-sphingosine. The conversion of the doubly labeled ceramide to 3-keto derivative is also described.


Determinations of adipose cell size and number in suspensions of isolated rat and human adipose cells. The osmic acid fixation-Coulter electronic counter method described for determining adipose cell size and number in intact adipose tissue fragments has been modified for use with suspensions of isolated rat and human adipose cells. Mean cell sizes in tissue fragments and isolated cell suspensions prepared from the same tissue are virtually identical in rats of various weights. No statistically significant difference in mean adipose cell size between tissue and isolated cell suspension was observed in human adipose tissue although the variability was much greater than in rat tissue. The distribution of cell sizes among replicate samples is more uniform in the isolated cell preparations, possibly reflecting the considerably larger quantities of tissue used in preparing isolated cells than in determining cell size and number directly from tissue fragments. An example of the utility of the modified method during routine metabolic studies with isolated rat epididymal adipose cells is described; isolated cells of increasing size can be obtained from rats of increasing body weight, or from the separated distal and proximal portions of the fat pads of rats of the same weight.


A novel semiautomated method for the estimation of free fatty acid in serum or plasma. A modification of the semiautomated assay method of Antonis (1965. J. Lipid Res. 6:307-312) for free fatty acid is presented. Free fatty acids are extracted from serum or plasma into di-n-butyl ether-2-methoxyethanol; the extract is almost free from phospholipids. The acids are analyzed in a portion of extract by a copper soap method using diphenylcarbazide. The extractant, being less dense than water, is easily separated from an aqueous phase both in the extraction of samples and in the assay of copper soaps. The assay is comparable in accuracy with well-tried titrimetric methods and is quicker and easier to operate.


Evaluation of Poly S-179 as a stationary phase for the gas-liquid chromatography-mass-spectrometry of bile acid methyl ester acetates. The stationary phase Poly S-179 has been found to offer distinct advantages over the previously reported SP-525 for the gas-liquid chromatographic separation of bile acid methyl ester acetates. Relative retention times of these bile acid derivatives are compared on the two phases.


Effects of adrenalectomy and prednisolone on the absorption and phosphorylation of thiamine in rats. After adrenalectomy, the absorption of [35S]thiamine in the small intestine of rats was significantly reduced. Injection of delta1,4-pregnadiene-11beta,17alpha,21-triol-3,20-dione (prednisolone) restored this absorption to some extent, but not to the value found in control (unoperated) rats. Treatment with 1% saline had no effect. Adrenalectomy also reduced the amount of [35S]thiamine that accumulated in the intestinal mucosa and liver compared with that found in controls, but treatment with prednisolone increased the retention of thiamine in these tissues. One per cent NaCl raised the retention of thiamine in the liver but not in the mucosa of the jejunum. The loss in body weight that occurred after adrenalectomy was not, however, corrected after treatment with either saline or steroid. In adrenalectomized rats, the concentrations of thiamine mono- and diphosphates in the intestinal mucosa and the liver were lower than in the control animals. The concentration of thiamine triphosphate was, however, higher in the intestinal mucosa after adrenalectomy, but remained unchanged in the liver. Treatment with prednisolone increased the concentrations of thiamine mono-, di- and triphosphates in both of these tissues in adrenalectomized rats, compared with unoperated control animals.


Urinary excretion of neurophysins in patients with kidney disease. The mean rate of excretion of neurophysins in the urine of 16 patients with kidney disease but without tubular dysfunction was 0.48 +/- 0.14 (S.E.M.) ng/min, whereas the rate in 16 patients with tubular dysfunction was 3.64 +/- 1.56 ng/min (significantly different; 2P less than 0.01). In the whole group of 32 patients there was a relationship (r = 0.57) between the rate of excretion of neurophysins in the urine and the clearance of lysozyme. The increased rate of urinary excretion of neurophysins observed in some patients with kidney disease therefore appears to be related to a disorder of renal tubular function. It is shown that the raised levels of neurophysins observed in the serum of some patients with kidney disease are not attributable to a decrease in the urinary clearance of neurophysins.


Preparation of antisera to three individual rat neurophysins and their use for radioimmunoassays. A number of antisera have been raised against individual rat neurophysins. One of these proved suitable for the specific radioimmunoassay of rat vasopressin-neurophysin and data validating such a use are presented. Use of this antiserum showed that vasopressin-neurophysin is present in rat neurohypophyses in equimolar amounts to vasopressin, while in the hypothalamus the assay detected twice as much neurophysin-like material as hormone. A number of partially specific antisera for rat oxytocin-neurophysin have been obtained and the best of these was raised against the minor neurophysin.


Response of chondrocytes isolated from human foetal cartilage to plasma somatomedin activity. Plasma somatomedin activity enhanced the incorporation of [3H]thymidine into chondrocytes isolated from human foetal cartilage during weeks 13-21 of gestation. Human growth hormone (0.1-20 muu./ml), human placental lactogen (0.1-5 microgram/ml) and insulin (0.25-10 muu./ml) had no direct effects on the synthesis of DNA in these chondrocytes, although insulin at concentrations of 2.5-100 mu./ml increased [3H]thymidine incorporation by up to 400%.