Nomenclature for factors of the HLA system. A Nomenclature Committee composed of geneticists and immunologists, including specialists in tissue typing, has met after each of the Histocompatibility Workshops beginning with the Third Workshop in 1967. The Committee, in part under the auspices of the World Health Organization and the International Union of Immunological Societies (WHO/IUIS), met after the Sixth Workshop in Aarhus in July 1975. The expanding knowledge of the genetics of the major histocompatibility system of man has necessitated a revision of the terminology for the HLA region following the principles established in previous reports. This has been done with as few changes as possible.


Distribution of the snail Biomphalaria glabrata, intermediate host of Schistosoma mansoni, within a St Lucian field habitat. A total of 6360 mud samples were obtained, in 62 collections made with an exhaustive sampling device, from banana drains on the West Indian island of St Lucia during fortnightly samplings over a 2(1/2)-year period. Analysis of counts of the snail Biomphalaria glabrata from these samples showed that this species had a contagious distribution. This finding is consistent with other evidence that banana drains form a rigorous habitat for B. glabrata. Its distribution was more contagious than that of Oncomelania quadrasi in certain Philippine habitats and several species of aquatic snail in various African irrigation canals. The exact transformation for normalizing the snail counts for standard statistical techniques was z = x(0.287) but the more convenient cube root transformation is probably adequate. However, if too few snails are collected (15 or fewer per 100 samples) or if the frequency distribution of snail counts is discontinuous, with too many widely separated high frequency counts, neither transformation will be entirely satisfactory.


Blinding and non-blinding trachoma: assessment of intensity of upper tarsal inflammatory disease and disabling lesions. A simplified method of evaluating both the intensity of active inflammatory disease and the visually disabling lesions of trachoma is presented. Evaluation of the active disease is based on the scoring of clinical signs, lymphoid follicles, and papillary hypertrophy. The irreversible lesions of trachoma are classified as conjunctival scarring, trichiasis and/or entropion, and corneal scarring.


Hepatitis B antigen, antigen subtypes, and hepatitis B antibody in normal subjects and patients with liver disease. The relative sensitivities of counterimmunoelectrophoresis (CIE) and haemagglutination assays for the detection of hepatitis B surface antigen (HB(s)Ag) and antibodies (anti-HB(s)) were compared. Twelve scientists from ten countries in Asia, Africa and the Pacific region participated in the study. The participants provided serum samples from 15 953 subjects comprising patients with acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC), as well as blood donors and other normal individuals. For the detection of HB(s)Ag in a reference panel serum, immune adherence haemagglutination (IAHA) was slightly more sensitive than passive haemagglutination inhibition (PHI); CIE was the least sensitive. Mean HB(s)Ag frequencies in patients with acute hepatitis, chronic hepatitis, cirrhosis, and HCC were significantly higher than in healthy controls. Passive haemagglutination (PHA) was more sensitive than CIE for the detection of anti-HB(s). The frequency of anti-HB(s) in patients with HCC was significantly lower than that in the other groups. Mean anti-HB(s) frequencies in patients with acute hepatitis, chronic hepatitis, and cirrhosis were not significantly different from that in normal subjects. Subtyping of HB(s)Ag was performed by PHI. Among asymptomatic carriers the predominant HB(s)Ag subtype in northeast Asia was adr.In India, ayw predominated in carriers, with the demarcation between adr and ayw occurring west of Burma. In West Africa the only subtype detected was ayw, but in East Africa the majority subtype was adw. The r subtype was found only in Asian populations east of India and in Western Pacific populations. In Papua New Guinea all four subtypes were identified. With one possible exception, the subtypes of HB(s)Ag-positive patients with liver disease reflected the predominant type in each geographic location.


Clinical observations on smallpox: a study of 1233 patients admitted to the Infectious Diseases Hospital, Calcutta, during 1973. The paper presents clinical observations on 1 233 persons with smallpox who were admitted to the Infectious Diseases Hospital, Calcutta, in 1973. The disease was of the modified type in 53 patients (4.3%), the ordinary type in 717 (58.2%), the flat type in 249 (20.2%), and the haemorrhagic type in 214 (17.3%). The fatality of these types of smallpox was found to be 5.7%, 26.8%, 88.4%, and 98.1%, respectively, and the overall case fatality was 50.7%. The haemorrhagic type was found mainly among older patients and affected males more often than females. The vaccination status of 1 218 patients was known. Of these, 901 (73.9%) were unvaccinated and had a fatality rate of 53.4%, whereas the 317 (26.1%) vaccinated patients had a fatality rate of 36.5%. Among the 201 haemorrhagic cases, 145 patients were unvaccinated (16.09% of the total number unvaccinated) and 56 (17.67%) had been vaccinated. Of 34 patients vaccinated during the incubation period, 19 (41.1%) died, whereas of 18 patients who had been vaccinated after the onset of fever, but before the appearance of rash, 9 (50%) died.


The minimum protective level of antibodies in smallpox. Blood samples from 57 contacts of 6 smallpox cases were tested for haemagglutination-inhibiting (HI) and neutralizing antibodies. All 6 contacts who subsequently developed smallpox were unvaccinated and had neutralizing antibody titres of 10 or less. However, 6 unvaccinated contacts with similar antibody levels did not develop smallpox. None of the 41 vaccinated contacts, regardless of their antibody level, contracted the disease.


A simple simulation model of tuberculosis epidemiology for use without large-scale computers. A large-scale computer service is not always available in many countries with tuberculosis problems needing epidemiological analysis. To facilitate work in such countries, a simple epidemiological model was made to calculate annual trends in the prevalence and incidence of tuberculosis and its infection, in tuberculosis mortality, and in BCG coverage, using average parameter values not specific for age groups or birth year cohorts. To test its approximation capabilities and limits, the model was applied to epidemiological data from Japan, where sufficient information was available from repeated nation-wide sample surveys and national statistics. The approximation was found to be satisfactory within certain limits. The model is best used with a desk-top computer, but the calculations can be performed with a small calculator or even by hand.


Prevalence of Salmonella serotypes in India: a 61-year study. During the period 1958-73, 8 027 strains of Salmonella were tested at the National Salmonella and Escherichia Centre: 3 834 strains from man, 3 018 from animals, 839 from sewage and water sources, and 336 of unknown origin. A total of 99 serotypes were identified: 47 from man, 83 from animals, and 35 from sewage and water sources. S. typhi was the commonest serotype in man, followed by S. weltevreden and S. paratyphi A. S. typhimurium was the commonest serotype isolated from animals, followed by S. weltevreden and S. anatum. In sewage and other water sources S. weltevreden was the commonest serotype identified followed by S. typhimurium and S. bareilly. Infection due to S. weltevreden in both man and animals increased considerably after 1970. Out of a total of 99 serotypes isolated from different sources, 13 were isolated from man only, 49 from animals only, 34 from both man and animals, 30 from both animals and water sources, and 22 from man, animals, and water sources.


Lactation and reproduction.

Lactation and reproduction. The authors review the literature on the effect of lactation on fertility in the absence of contraception and on the effects of contraceptive measures on lactation. They examine data from several countries on the intervals between births and on the return of menstruation and ovulation after childbirth, comparing lactating with nonlactating women. They conclude that lactation is an inefficient contraceptive for the individual, but that in populations sustained lactation is associated with reduced fertility. Possible physiological mechanisms causing lactation amenorrhoea are discussed. Though much of the literature on the effect of contraceptives on lactation is inadequate, there is general agreement that the estrogen component of hormonal preparations has an adverse effect on lactation, but that progestins alone do not. Many questions remain. Is this effect seen in established lactation, or only in the puerperal period? Is it a direct pharmacological effect, or are pill-users the mothers least motivated to maintain breast-feeding? Does a close relationship exist between hormones given and lactation performance? The authors comment on some of the technical deficiencies of previous studies in this field and discuss practical possibilities of, and limitations to, obtaining adequate scientific information in the future.


Testing the susceptibility of Simulium damnosum (Diptera: Simuliidae) to insecticides in the laboratory and in the field. There are several methods for measuring the susceptibility of blackfly larvae to insecticides. In 1970 the provisional method suggested by the World Health Organization was used with a view to establishing baseline data on the susceptibility of S. damnosum larvae to insecticides; this method was found to be unsuitable. Thereafter, attempts were made to develop a reliable susceptibility testing method for S. damnosum. This report describes the method that yielded the best results in the laboratory; a second method suitable for use in the field is also described. The results from several replicates of such tests gave reasonably straight log-dosage-probit-mortality curves from which values of LC(50) could be calculated. The overall control mortality in the laboratory series of tests was about 9% and that in the field series was about 4%.


The use of psychotropic drugs in developing countries. Psychotropic drug therapy combined with other forms of treatment provides an effective means for the control and treatment of a number of mental disorders. In developing countries a wide range of health workers must be prepared to use psychotropic drugs if there is to be a significant improvement in mental health care. A number of problems are involved: not all mental disorders respond to drug treatment; the range of available drugs is very wide; side effects are relatively common; patients may not take prescribed drugs regularly; and there are dangers of overuse, abuse, and overdose. Some of the problems could be overcome by: (a) focusing on a limited number of conditions of public health importance; (b) establishing a clear policy as to which drugs should be available at various points in the health service and limiting the range of such drugs; (c) adopting a more flexible system of task sharing in psychotropic drug therapy; (d) coordinating training programmes; and (e) setting up a central policy body concerned with mental health in health ministries.


Evaluation of the impact of various diseases on mortality. The scarcity of resources within the health sector creates serious allocation problems. If it were legitimate to allocate the available means to the various diseases according to their impact on the length of human life, how should this impact be evaluated? That is the purpose of this study, which compares and evaluates methods for measuring the importance of various diseases from the point of view of their lethal effects and significance for life expectancy.THREE METHODS OF EVALUATION HAVE BEEN USED: the percentage of all deaths due to the disease (method A); the gain in life expectancy that would occur if the disease were eradicated as a cause of death (method B); and the change in the ratio between productive and nonproductive groups that would result from eliminating the disease as a cause of death (method C). The analyses are based on the total number of deaths in Denmark in 1969 and 1971. According to (A), one-third of all deaths was caused by heart diseases, one-fourth by cancer, and one-tenth by stroke. The results of method (B) were in agreement with those of method (A); the greatest gain in life expectancy was obtained by eliminating cardiac diseases; cancer ranked second, and accidents third for men and fourth for women. Method (C) yielded contrasting results. The result of eliminating most diseases would be a decrease in the proportion of persons of productive age. This was most distinct for cardiac diseases. The most important exception to this rule was accidents: if these were eliminated, the distribution of the population in the productive and nonproductive age groups would remain stable.


Respiratory virus disease in Malaysian children: a serological study. Paired sera from 101 Malaysian children aged up to 10 years and suffering from respiratory illnesses were examined serologically for evidence of respiratory viral infections. Of these children, 32.6% showed rising antibody titres for one or more of the test agents. Respiratory syncytial virus appeared to be the main respiratory pathogen involved, followed by Mycoplasma pneumoniae, parainfluenza viruses, adenoviruses, and influenza A virus. These findings are generally similar to those reported by others in temperate and tropical countries.


Abolition of ischemic response to atrial pacing following aortocoronary bypass surgery. To determine the effect of successful aortocoronary bypass surgery (ACBS) on left ventricular (LV) function, the ischemic response to right atrial pacing (RAP) was studied in 22 angina patients before and 3.8 +/- 1.1 months after surgery. All patients were free of angina after ACBS and had at least one patent graft. Before ACBS, RAP induced angina in 15 patients (Group 1) but not in 7 patients (Group 2). After ACBS, no patient had angina with RAP despite the increased maximum rate of pacing. Post-pacing LV end-diastolic pressure (LVEDP) after ACBS decreased in Group 1 from 25 +/- 6 to 15 +/- 6 mm Hg (P less than 0.01), but not in Group 2. Changes in ejection fraction, cardiac output, resting LVEDP, or LVEDP after LV angiography were not significant in either group and were therefore not useful in evaluating the result of ACBS. However the ischemic response to right atrial pacing was abolished by successful aortocoronary bypass surgery, suggesting improved myocardial perfusion during stress.


Continuous rapid atrial pacing to control recurrent or sustained supraventricular tachycardias following open heart surgery. A technique is described to control recurrent or sustained supraventricular tachycardia associated with rapid ventricular rates following open heart surgery. The technique utilizes a pair of temporarily implanted atrial epicardial wire electrodes to pace the heart. In one group of patients with recurrent atrial flutter and 2:1 A-V conduction, continuous rapid atrial pacing at 450 beats/min produced and sustained atrial fibrillation. The ventricular response rate immediately slowed when compared to that during atrial flutter, and if further slowing was required, it was easily accomplished by the administration of digitalis. Another group of patients with different arrhythmias (recurrent paroxysmal atrial tachycardia, sustained ectopic atrial tachycardia, or sinus rhythm with premature atrial beats which precipitated runs of atrial fibrillation) was treated with continuous rapid atrial pacing to produce 2:1 A-V block. In all instances, the continuous rapid atrial pacing suppressed the supraventricular tachycardia and maintained the ventricular response rate in a therapeutically desirable range. It was demonstrated that the technique is safe, effective, and reliable.


Liver disease--a prominent cause of serum IgE elevation. Serum IgE concentrations were elevated in thirty-seven out of sixty-seven patients (55%) with acute or chronic liver disease of widely differing aetiology. The mean IgE concentrations in these patients showed an eight-fold increase above that observed in control subjects. Increased IgE levels in patients with liver disease occurred in the absence of eosinophilia, clinical evidence of atopy or other known causes of IgE elevation. No IgE-containing plasma cells were detected in the liver biopsies from thirty-two of the sixty-seven patients tested. Peripheral blood T cells were significantly decreased from normal in the patients with liver disease, but no correlation emerged between serum IgE levels and absolute peripheral blood T-cell numbers. These findings emphasize the importance of liver disease as a significant cause of serum IgE elevation.


T and B lymphocytes in South American pemphigus foliaceus. T and B lymphocytes were quantified in peripheral blood of thirty patients with South American pemphigus foliaceus according to their ability to form rosettes with sheep erythrocytes (E) or sheep erythrocytes sensitized with antibody an complement (EAC). When compared with the counts obtained from thirty normal subjects, a decrease was found in the total T-lymphocyte count (32-33+/-7-7 versus 46-7+/-8-7) and in the T functional lymphocyte count as detected by the active rosette test. The mean percentage of B lymphocytes within the total number of lymphocytes was not significantly different from that of normal subjects (24-0 +/- 8-2 versus 25-3+/-8-0). Lymph node sections from three pemphigus patients examined for E or EAC adherence showed depletion of T cells in the paracortical areas. The low percentage of E cells in the peripheral blood and the depletion of E cells in paracortical areas of lymph nodes from patients with South American pemphigus foliaceus may reflect an impaired cellular immunity.


Stimulation of lymphocytes by antigen in microplate cultures; absence of an effect of transfer factor in vitro. The best conditions for the optimum stimulation of human leucocytes by antigens, including protein purified derivative (PPD), streptokinase-streptodornase varidase (SKSD) and tetanus toxoid have been studied in microplate cultures. The leucocytes of each donor have their own characteristic response to antigen which depends on the culturing conditions such as antigen concentrations, cell concentrations and the time of measuring the rate of DNA synthesis. Thus, no conditions provide a universal optimum for antigen stimulation in vitro. Leucocyte dialysates, i.e. potential transfer factors, have been prepared from donors, who between them are strongly positive to the antigens PPD, SKSD, tetanus toxoid, diphtheria toxoid and Keyhole limpet haemocyanin. In contrast to some previous reports these leucocyte dialysates had no effect on the thymidine incorporation by leucocytes grown in the presence of these antigens. It is suggested that the selection of optimal conditions for the response to antigen may have obscured the effect of any non-specific enhancement of reactivity b lyeucocyte dialysates.


The effect of cyclophosphamide and irradiation on cells which suppress contact sensitivity in the mouse. Contact sensitivity was produced in mice by painting the skin with picryl chloride and was assessed by the increase in ear thickness following local challenge. Contact sensitivity was passively transferred by immune lymph node and spleen cells taken at 4 days. The mice were then challenged immediately and the reactions read at 24 and 48 hr. Immune lymph node and spleen cells taken at day 8 virtually fail to transfer. Experiment showed that they contain cells which suppress passive transfer. These are demonstrated by mixing approximately equal numbers of 4-day cells, which transfer contact sensitivity, and cells taken at later times and injecting them intravenously into recipients. These 'suppressor cells' can be demonstrated by day 6 and are still present at day 11 after immunization. The precursors of the suppressor cells are sensitive to cyclophosphamide. Irradiation of immune mice 2 days before taking cells also selectively inactivates the suppressor cells. When mice are pretreated with cyclophosphamide before immunization or irradiated 2 days before transfer, the lymph node and spleen cells taken on day 9 after immunization transfer contact sensitivity. In contrast the same number of cells from untreated mice were inactive. This suggests that the cells which mediate passive transfer or their precursors may occur in an inhibited form in lymph nodes and spleen at later times after immunization. These suppressor cells in immune mice differ from the T suppressor cells produced by the injection of picryl sulphonic acid--an agent which causes unresponsiveness: (1) the precursors of the T suppressor cells resist cyclophosphamide; (2) the T suppressor cells are found in mice treated so as to produce unresponsiveness while the other type of suppressor cells occurs in mice immunized for contact sensitivity. However, both types of suppressor cells are selectively inactivated by irradiation as compared with the cells which mediate contact sensitivity and both are able to act on the effector stage of contact sensitivity.


Antibodies for T lymphocytes in systemic lupus erythematosus. Sera of twenty-four patients with systemic lupus erythematosus were evaluated for antibodies cytotoxic for autologous lymphocytes. Such antibodies were domenstrable in twenty-two of these sera, whereas only one out of twenty patients with other connective tissue or lymphoproliferative disease showed a positive test. The antibodies remained detectable even when the patients went into remission. Sera containing the lymphocytotoxic antibodies were tested on cell fractions enriched for T or B lymphocytes. Primarily T lumphocytes wree affected by these antibodies. The presence of antibodies cytotoxic for T lymphocytes corresponded with a deficit of circulating T lymphocytes observed in most of our patients with systemic lupus erythematosys.


Soluble factors in immune sera of mice. II. Reversible loss of surface immunoglobulin induced in vitro. Serum collected from BALB/c mice at different time intervals during primary immunization induces in vitro a decrease of Ig+ cells in short-term cultures of normal spleen cells. The decrease was shown to be caused by loss of surface immunoglobulin (Ig) for the following reasons: no cell loss was detected which coulc account for it; the number of Ig+ cells returned to normal levels when the serum was removed and the cells were cultured further in fresh medium. The serum activity was recovered in a fraction with molecular weight less than 10,000 Daltons. The factor was active at 4 degrees C and on cells treated with high doses of Con A. Only a portion (25-30%) of the Ig+ cells are affected by this factor since after the initial decrease no further changes were observed during the 7 hr culture period. No changes in the distrubution of surface Ig was detected by fluorescent techniques on the remaining Ig+ cells. This factor was detected as early as 6 hr after immunization but its concentration was found 4-fold higher on the 7th day.


Suppressed hetergeneous antinuclear antibody response in lymphomabearing NZB/NZW mice. A long-term study of cyclophophamide-induced suppression of auto-immune disease in NZB/NZW mice demonstrated the oncogenic potential of this alkylating agent. In seven of ten mice dying with lymphomas, retrospective examination of serial tests for heterogeneous ANA showed terminal decreases of titres or reversion of tests to negative. Suppressed titres were found in one of seven mice dying with other tumours and in four of twenty-nine mice succumbing to renal disease-vasculitis. Transplantation studies confirmed the association between growing lymphoma tissue and decreasing ANA titres. In the immunosuppressed host with connective tissue disease, the loss of certain auto-antibodies may signal development of a lymphoreticular neoplasm. Tumour-induced impairment of auto-immune responsiveness may reflect the loss of specific ANA that fulfil a protective role and inhibit neoplastic growth.


Mechanisms of corticosteroid action on lymphocyte subpopulations. II. Differential effects of in vivo hydrocortisone, prednisone and dexamethasone on in vitro expression of lymphocyte function. The present study was undertaken to determine what, if any, differential effect various commonly used corticosteroid preparations had on the numbers and specific functions of lymphocyte subpopulations when these agents were administered in equivalent pharmacological dosages. Normal volunteers received a single dose of either 320 mg of hydrocortisone intravenously, 80 mg of prednisone orally, or 12 mg of dexamethasone orally. There was a marked lymphocytopenia and monocytopenia maximal 4-6 hr following administration of all three corticosteroid preparations with almost identical kinetics and degree of fall in total cell numbers as well as proportions of thymus-derived and bone marrow-derived lymphocytes. Hydrocortisone and prednisone caused only a slight suppression of phytohaemagglutiinin (PHA) induced lymphocyte blastogensis which could be reversed at supra-optimal concentrations of PHA. On the contrary, dexamethasone administration casued a marked suppression of PHA responses which was not reversed by supra-optimal PHA stimulation. In addition, hydrocortisone and prednisone administration did not suppress non-specific PHA-induced cellular cytotoxcity, while dexamethasone caused a marked suppression (P less than 0.001) of cytotoxicity. These studies show that although equivalent anti-inflammatory doses of these three corticosteroid preparations cause almost identical suppression of the numbers of circulating lymphocyte populations, they have a differential effect of the numbers of circulating lymphocyte populations, they have a differential effect on a certain in vitro functional correlates of cell-mediated immunity.


Respiratory involvement in systemic lupus erythematosus. A clinical and immunological study. Thirty patients fulfilling conventional criteria for systemic lupus erythematosus and who presented with extensive pleural and pulmonary involvement were studied retrospectively. Four overlapping patterns of respiratory disease were identified and observations were made on their clinical presentation, radiographic abnormalities and response to treatment. A low incidence of severe renal disease was found in this series of patients and this was in keeping with the general finding of low serum binding using native DNA in a globulin Farr-binding technique (greater than 20% binding in only 4/21 (19%) of the series) and normal or elevated serum complement (C3) levels. Precipitating antibody detected by double diffusion and counter-current immunoelectrophoresis and probably reacting in most cases with single-stranded DNA was, however, detected in 66% of pretreatment serum samples tested. This evidence supports the idea that different types of anti-nuclear antibody may be associated with different clinical manifestations seen within a group of patients who broadly fulfil the criteria for SLE.


Measles antibodies and autoantibodies in autoimmune disorders. Measles CF antibodies have been examined in the sera of patients with a variety of clinical disorders associated with the production of autoantibodies. Previous reports of high-titre reactions in DLE and chronic active hepatitis have been confirmed, the titres in the latter disorder being particularly elevated. Mean antibody titres to measles in patients with rheumatoid arthritis were significantly lower than in matched controls, and an inverse correlation between measles antibody levels and serum globulin levels was found. Measles antibody titres in patients with myasthenia gravis and primary biliary cirrhosis did not differ significantly from those found in controls. However, subdivision of patients with rheumatoid arthritis, myasthenia gravis and primary biliary cirrhosis showed that the presence of anti-nuclear antibody (ANA) was associated with significantly increased measles antibody levels compared with the ANA-negative sera. The presence of gastric parietal cell antibody or thyroid microsomal antibody did not appear to be associated with increased measles antibody levels, whether or not they occurred in association with previous anaemia or thyroid disease. Possible explanations for these findings in terms of immune complex formation and immune hyper-reactivity are discussed.


The effect of Corynebacterium parvum on the humoral and cellular immune systems in patients with breast cancer. Corynebacterium parvum, a Gram-positive anaerobic bacillus thought to be a strong immunological stimulant, has been shown to decrease tumour growth and prolong survival in patients with metastatic disease. Study of the effect of a single injection of a strain of C. parvum (CN. 6134) in six patients with stage IV metastatic breast cancer is reported. Results of laboratory tests to judge the physical and immunological effects of the drug infusion 24 hr post-treatment and weekly thereafter for 3 weeks are evaluated. Within 24 hr after C. parvum administration, most patients experienced fever and nausea. Blood counts and differential counts exhibited increased values 24 hr after treatment with a strong shift to the left. Lymphocyte and monocyte counts were greatly depressed at 24 hr. T-cell numbers in peripheral blood did not appear to be altered, but the picture with regard to B cells was less clear. Normal count was recovered by day 8. It appears that intravenous administration of C. parvum produces a temporary marked immunological depression which returns to essentially normal values in 8 days. The return to normal may be accompanied by resolution of the endotoxin-like syndrome of side-effects. Further study of patients receiving this therapeutic agent is important to detect enhancement of the anti-tumour immunological response precipitated.


Depression of the T-lymphocyte response to phytohaemagglutinin by renal cells. The lymphocytic infiltrate in the renal parenchyma is a consistent histological feature of pyelonephritis, but the role of the lymphocytes in the immunobiology of pyelonephritis is not known. In this investigation the influence of the local environment on the potential function of T lymphocytes in the kidney was investigated. The experiments have demonstrated that the response of rat lymphocytes to stimulation in vitro with phytohaemagglutinin (PHA) can be entirely ablated by normal kidney cells. Even when the number of kidney cells added to cultures of lymphocytes was less than 2% of the cells present some ablation of T-lymphocyte function could be detected. The biological characteristics of the factor causing ablation of the PHA responsiveness of T lymphocytes were partially characterized and the factor appears to have unique features that differentiate it from lymphocyte chalones and other tissue factors influencing lymphocyte function. The results may explain recent findings where T lymphocytes were found to be the predominant lymphocyte in the inflammatory infiltrate but were not responsive to PHA in vitro.


Intestinal blood loss after a new anti-inflammatory drug, sulindac. Sulindac was tested for fecal blood loss in 40 healthy male subjects whose red cells had been labeled with Na2 51 CrO4. Two daily dose levels of 240 mg and 400 mg were compared with 4.8 gm of aspirin and placebo for 2 wk. At day 15, aspirin-induced blood loss was greater than that of both dose levels of sulindac and of placebo (p less than 0.05). There were no significant differences between the two sulindac groups and the placebo group. Aspirin caused more adverse reactions than sulindac, 240 mg (p less than 0.05), 400 mg (p less than 0.05), and placebo (p less than 0.05).


Fiberoptic colonoscopy: complications of colonoscopy and polypectomy. Members of the American Society of Colon and Rectal Surgeons were polled regarding complications of colonoscopy and polypectomy. Sixty per cent of the members responded. One hundred sixty-two surgeons were found to be performing colonoscopy, and this paper presents the accumulated data on complications. A total of 20,139 colonoscopies were performed, with 12,746 diagnostic procedures and 7,393 polypectomies. The overall complication rate was 0.4 per cent with diagnostic colonoscopy and 1.8 per cent with polypectomy. Based upon these complications, guidelines in regard to history, preparation, and technique of colonoscopy and polypectomy are discussed.


Cyproterone-mediated stimulation of delta-aminolevulinic acid synthetase in chick embryo liver cells. In cultured chick embryo liver cells, cyproterone and cyproterone acetate, synthetic anti-androgenic steroids, were found to be potent inducers of mitochondrial delta-aminolevulinic acid (ALA) synthetase, the rate-limiting enzyme in the heme biosynthetic pathway. Both steroids produced a detectable increase in enzyme activity at 5 muM and a maximal stimulation, 36-fold for cyproterone and 29-fold for cyproterone acetate, at 55 muM. The dose-response curves of the steroids differed, however, in that cyproterone acetate produced a greater mean stimulation of the enzyme at concentrations less than approximately 25 muM, whereas, at higher concentrations, cyproterone was the more effective inducer. Increased activity of ALA synthetase was not apparent until about 12 hours after the addition of cyproterone, and maximal activity was not achieved until 20-24 hours. The induction of ALA synthetase by these anti-androgens was prevented by actinomycin D, cordycepin, anisomycin, cycloheximide, and puromycin. These results suggest that new RNA and protein synthesis are necessary for enzyme induction. The cyproterone-mediated induction of the enzyme was inhibited 50% by 2 muM heme, the putative physiological inhibitor of ALA synthetase. These antiandrogens, unlike other potent steroid inducers of this enzyme in chick embryo liver, do not possess either a 5beta-pregnane or 5beta-androstane nucleus. The stimulation of hepatic ALA synthetase represents the first example of a direct effect of these steroids on enzyme induction.


[Charge-transfer complexes of hematoporphyrin with pteridine derivatives]. Charge-transfer complexes of hematoporphyrin with pteridine derivatives (folic acid, folinic acid, methotrexate) were studied in solutions buffered at pH 7.5 - 9 -10. The association constants for folic acid are pH dependent; at pH 7.5 the constant is 1240 wheras at pH 10 it is only 65. The association constant for folinic acid was determined only at pH 7.5 and was found to be 55. The association constants of methotexate are approximately the same at different pH's.


Clones of B lymphocytes: their natural selection and expansion. The operation of clonal selection for cells of the B-lymphocyte line is discussed with regard to: 1) The clonal repertoire determined by antigen binding to B lymphocytes, which is much larger than that determined by limiting dilution cloning assays. This quantitative difference is interpreted in terms of the multiple shared specificities of each antibody molecule. 2) Multiclonal responses and initial selection by antigen of particular clones (preferential primary selection). 3) Clonal dominance. During an immune response one clone (or a small number of clones) of B cells is preferentially selected and proliferated, apparently at random, from a heterogeneous population of cells capable of responding to the given antigen. Co-dominance of two or more clones simultaneously can be obtained by mixing selected clones. Secreted antibody is seen as playing a role in the establishment of clonal dominance. A model for clonal expansion is presented. The model attempts to explain the generation of memory and antibody secreting cells within each clonal expansion in terms of the ratio of two signals, one for proliferation and one for differentiation. The delivery of these signals is proposed to involve the receptor antibody-antigen interaction for proliferation and a self-recognition site interaction for differentiation.


Studies on the mode of action of ecdysterone in adult female Aedes aegypti. Injection of ecdysterone into non-blood fed adult female Aedes aegypti results in a marked stimulation of aromatic-L-amino-acid decarboxylase (formerly DOPA-decarboxylase) activity (Schlaeger and Fuchs, 1974a). When the hormone and alpha-amanitin are injected either simultaneously or if the toxin is administered first no inhibition of subsequent enzymatic activity is observed and in fact substantial enhancement occurs. Cordycepin injection along with ecdysterone gives results similar to alpha-amanitin. The inhibitors by themselves elicit a very small increase in DOPA decarboxylase activity compared to saline-injected controls. Conversely, actinomycin D causes severe depression of ecdysterone-mediated DOPA decarboxylase activity as dose cycloheximide and puromycin. We interpret our data to mean that the mRNA for DOPA decarboxylase is already present prior to exposure to ecdysterone. We postulate that the function of the hormone would be to modulate translation of specific pre-formed mRNA's by an unknown mechanism or to induce transcription of specific tRNAs necessary for the initation of translation of selecting existing messengers.


Does lower esophageal sphincter incompetency contribute to esophageal bleeding? The purpose of this study is to determine whether lower esophageal sphincter (LES) incompetency is a common occurrence in patients with liver cirrhosis and contributes to the development of variceal bleeding. Resting LES pressure (17.8 +/- 1.1 mm Hg) in 35 patients with cirrhosis was similar to that of our control population (17.3 +/- 2.0 mm Hg). No differences were found among patients with ascites, variceal hemorrhage, or with different degrees of hepatic decompensation. In both patients and control subjects the LES responded with a significant pressure increase to gastric alkalinization. Symptoms and radiological evidence of gastroesophageal reflux were extremely uncommon in patients with liver cirrhosis. Based on these data it is unlikely that acid-pepsin regurgitation is a significant factor in the development of variceal hemorrhage.


Indications for surgery in Crohn's disease: analysis of 500 cases. Data for 500 patients with Crohn's disease who underwent operations were analyzed (316 patients, 1966 to 1969; 184 patients, 1972 to 1973) by comparison of various anatomic disease locations (clinical pattern): (1) ileocolic, 225 patients; (2) small intestinal, 130 patients; (3) colonic, 127 patients; (4) anorectal, 18 patients. Indications for surgery were tabulated and compared using statistical analysis for the three large patterns. For patients with ileocolic Crohn's disease, the primary surgical indications were internal fistula and abscess, 44%, intestinal obstruction, 35%, and perianal disease, 12%. For patients with Crohn's disease of the small intestine, the primary surgical indications were intestinal obstruction, 55%, and intestinal fistula and abscess, 32%. Patients with colonic Crohn's disease had a significantly more diverse surgical indication, with poor response to medical therapy, 26%, internal fistula and abscess, 23%, toxic megacolon, 20%, and perianal disease, 19%. These values were highly statistically significant (P less than 0.0001) in all instances but one. This study demonstrates that statistically significant differences occur in the surgical indication depending on the location of Crohn's disease. Patients with ileocolic, small intestinal, and colonic involvement have striking differences in clinical course. It is concluded that Crohn's disease is not a homogeneous entity, but should be recognized as having a varying course depending on clinical pattern.


Immunological studies in familial primary biliary cirrhosis. Tests for cell-mediated immunity and presence of autoantibodies were performed in a mother and daughter with primary biliary cirrhosis. Lymphocytes transformation to phytohemagglutinin, delayed cutaneous response to one or more skin test antigens, and percentage of peripheral T and B lymphocytes were normal in both patients. Although successful in the mother, dinitrochlorbenzene sensitization was not achieved in the daughter. Histocompatibility antigens 1 and 8, elevated levels of IgM, and antibodies to mitochondria, smooth muscle, and skeletal muscle were present in both patients. However, the clinical course was more severe in the daughter who developed portal hypertension with bleeding esophageal varices requiring portacaval anastomosis. Except for intermittent pruritus, the mother has remained asymptomatic.


Massive splenic infarction in cirrhosis: report of a case with spontaneous disappearance of hypersplenism. A cirrhotic patient with massive splenic infarction is described. Celiac angiography showed normally opacified splenic artery and vein and a markedly enlarged spleen with large avascular zones. Splenic infarction was associated with the spontaneous disappearance of a syndrome of hypersplenism. The spleen was surgically removed. Histological examination showed multiple thromboses of the small arterial and venous vessels. The cause of this infarct remained unclear.


The subpopulations of circulating white blood cells in inflammatory bowel disease. A detailed analysis of the species of lymphocytes was carried out in 58 patients with inflammatory bowel disease (IBD). These individuals were further divided into 31 with Crohn's disease (CD) and 27 with ulcerative colitis (UC). There were 13 CD patients with only small bowel involvement called "regional enteritis" and 18 who had some degree of colonic involvement called "ileocolitis". Similarly, the UC group was subdivided into 9 patients with disease confined to the rectosigmoid area called "proctosigmoiditis" and 18 with more extensive involvement called "universal colitis". We also studied 13 patients who had undergone previous colectomy and ileostomy and 78 healthy age- and sex-matched controls. Although there was no increase in the absolute number of lymphocytes in patients with ileocolitis and universal colitis, the percentage of these cells was decreased because of an increase in both polymorphonuclear leukocytes and monocytes. In IBD and its subgroups, mean T lymphocytes, determined by the sheep red blood cell rosette technique, were not significantly different from the controls either in percentage or absolute number. Furthermore, no difference was noted between UC and CD. However, there seems to be a subpopulation of patients with UC or CD whose T cells are reduced below 1 SD of the mean. There was also no difference in the number of immunoglobulin-bearing B cells in both diseases; however, when the B cells were enumerated by their ability to rosette with antibody-complement-coated sheep cells (EAC), we found a marked decrease in percentage (P less than 0.001) and absolute number (P less than 0.0005) relative to the control population. The decrease bore a direct relation to the severity of the disease process and, although more marked in patients with UC, was present in CD also.


Adhesion of human blood platelets to glass polymer surfaces. II. Demonstration of the presence of a natural platelet adhesion inhibitor in plasma and serum. Adhesiveness of washed platelets resuspended in citrated plasma, serum, or several different media has been investigated. A method specific for quanititation of adhesion was used. Platelets suspended in saline of Tyrode's solution were found to be highly adhesive to glass, polyethylene, polyvinyl chloride, or Cuprophane. This adhesiveness of platelets to test surfaces decreased by nearly 50% when plasma was the suspension medium. When the suspension medium was serum, the decrease in adhesion was nearly 75%. Cohn fraction V also decreased the adhesiveness of platelets significantly, but highly purfied albumin had only a small effect. Several pharmacologic agents decreased platelet adhesiveness when added to platelets suspended in plasma or serum, but had negligible effect on the adhesiveness of platelets suspended in artificial media devoid of proteins. Normal washed platelets, when suspended in citrated plasma obtained from an afibrinogenemic donor or in normal serum, showed a significant decrease in adhesion compared to the same platelets suspended in normal citrated plasma. Addition of fibrinogen to afibrinogenemic plasma or normal serum restored the adhesiveness of platelets to normal levels. Normal platelets resuspended in plasma obtained from a thrombasthenic donor exhibited normal adhesiveness. These observations suggested that while fibrinogen promotes platelet adhesion, plasma or serum possess also an adhesion-inhibiting activity.


[Sex difference in the circadian rhythm of corticotropin-releasing activity in the rat hypothalamus (author's transl)]. Sex difference in the circadian rhythm of the hypothalamic content of Corticotropin-Releasing Factor (CRF) and plasma corticosterone levels was examined in the rat. In the male rat the CRF content was higher in the afternoon than in the morning. In contrast, the hypothalamic CRF content in the female rat was higher in the morning. The peak value was found at 8 a.m. and it fell rapidly around noon with a concomitant sharp rise in the plasma corticosterone. Thus, the whole pattern of CRF content during a 24-hour period makes a mirror image of that of plasma corticosterone. The influence of gonadal hormones on the sex difference was then examined by following variations in the CRF content after ovariectomy. Even after chronic ovariectomy, essential features of the female pattern of CRF rhythm persisted: rapid fall of the CRF content was accompanied by a sharp rise in the plasma corticosterone level. It is noteworthy however that ovariectomy reduced the morning level of CRF activity without elevating the afternoon level, resulting in a peak shift toward noon. The female pattern of CRF rhythmpersisted similarly after bilateral ganglionectomy which is known to affect the biogenic amine rhythm in the pineal gland. The persistence of the female pattern may suggest some endogenous nature of the rhythm. It was then possibly that sex differentiation in the central nervous system (CNS) might be related with manifestation of the female pattern. In order to elucidate this point, postnatal development of circadian rhythm of CRF activity was examined in male and female rats separately. It was found that the CRF rhythm became manifest both in male and female rats around the third week of postnatal life and there was no essential difference in their rhythm pattern. In other words, both male and female rats begin their CRF rhythm with so-called male pattern, with higher values in the afternoon than in the end of the third week of life. In contrast, the CRF rhythm in females rats showed a marked change at ages of five to six weeks during which the onset of puberty intervened in our series of experiment;during this period, the CRF rhythm in female rats turned into the so-called female pattern, with higher values of CRF activity in the morning than in the afternoon. It was of interest that a marked rise in the plasma corticosterone, characteristic of mature female rats, concomitantly appeared. Further attempt were made to examine the effect of gonadal hormones in the periratal period. The hypothalamic CRF content in androgen sterilized female rats as well as in neonatally castrated male rats showed no circadian rhythm. The results implicate subtleness of hormonal effect in simulating physiological processes. In fact, CRF rhythms are variable depending on stages of the estrous cycle: during proestrus and estrus, the CRF content was markedly higher in the morning (9 a.m.) than in the afternoon (4 p.m.). But no significant difference was observed between them during diestrus I and II...


In vitro effects of zinc chloride on spontaneous sheep red blood cell (e) rosette formation by lymphocytes from cancer patients and normal subjects. The in vitro spontaneous formation of sheep red blood cell (SRBC) (E) rosettes by peripheral blood lymphocytes is altered in vitro by the addition of micro-molar quantities of zinc chloride. Appropriate concentrations of zinc chloride cause marked enhancement of rosette formation, both in normal subjects and in cancer patients with low initial E rosette values. The effect is dependent upon zinc treatment of the lymphocyte and not the SRBC. Other zinc-induced effects on E rosettes include retardation of temperature-dependent spontaneous decay, increased mechanical stability due to enhanced of "capping." The formation of EAC rosettes is not altered by the presence of zinc.


Distribution of T and B lymphocytes in lymphoid tissue of infants and children. Normal lymphoid tissue from children undergoing elective surgery was examined for T and B lymphocyte distribution. Although established for peripheral blood and bone marrow, T and B lymphocyte distributions have not been previously reported for lymph nodes, appendix, thymus, and spleen tissues in children. Thymus-dependent T cells were determined by the sheep erythrocyte rosette technique, and thymus-independent B cells were determined by the fluorescent labeling of surface immunoglobulins A (IgA), G (IgG), and M (IgM). Fifty percent of lymph node cells were either T or B cells; 65% of these cells were T lymphocytes, whereas 58% of B cells were of the IgM subclass. Less than half of the appendix cells were either T or B cells; 47% of these were T lymphocytes, and the remainder B lymphocytes had subclass distribution similar to that of lymph nodes but different from peripheral blood and bone marrow where B cells bearing IgG predominate. Thymus tissue contained 43% T cells and less than 1% B cells, but the spleen was composed largely of B cells, predominantly of the IgM type. Lymphoid tissue from nine children with either inflammatory or neoplastic diseases were studied and included for contrast. This paper establishes relative distribution values for T and B lymphocytes in normal lymphoid tissue and points out the potential use of this technique to quantitate deviations from normal in certain inflammatory and neoplastic diseases.


Tube leukocyte adherence inhibition assay for the detection of anti-tumor immunity. I. Monocyte is the reactive cell. Peripheral blood leukocytes (PBL) from breast cancer patients with early or localized cancer fail to adhere to glass in the presence of breast cancer extract. The same leukocytes do not react to unrelated tumour extracts. Enrichment and depletion of certain PBL populations from patients with apparently localized breast cancer indicated that the indicator and/or reactive cell manifesting non-adherence in the presence of appropriate tumour antigen was phagocytic, glass adherent in the absence of tumour antigen and had cell surface Fc-receptors. The cell involved, therefore, appears to be the circulating monocyte. These results show that in the tube LAI assay, the peripheral blood monocyte appears to react directly with the tumour antigen resulting in a loss of its property of adherence to glass.


Sulphydryl and disulphide groups in normal and irradiated epidermis. The distribution of sulphydryl (SH) and disulphide (SS) groups in normal and irradiated epidermis of newly-born rats was studied by microspectrophotometric quantitation of the Mercury-Orange reaction. In order to obtain information on (1) the total content of both groups within the different strata of the epithelium, and (2) the mean SH and SS concentration, which is an indicator of the density of these groups per unit of tissue. The content of both groups increased in relation to the extent of irradiated area, whereas there was a decrease in the mean concentration. The increase in content of SH and SS would lead to augmented production of keratin substance previously reported in irradiated epithelia, coincidentally with an acanthotic response. The decrease in concentration of SH and SS indicates a destruction of these groups at cellular level, which may be due to membrane alterations. The findings would explain an acceleration of the keratinizing process together with modifications, both in quality and in quantity, in the horny substance.


Somatic aberration induction in Tradescantia occidentalis by neutrons, x- and gamma-radiations. I. Dosimetry. The dosimetry is described for an investigation of the induction of somatic aberrations in Tradescantia occidentalis by substantially mono-energetic neutrons in the energy range 100 keV to 15 MeV, by 200 keV X-rays and cobalt-60 gamma-radiation. Spectrometry was carried out for both neutrons and X-rays. Neutron fluence was measured by uranium fission chambers. Two types of ionization chamber were employed for dose measurement. One chamber was manufactured of CH-plastic and filled with acetylene and the other of graphite and filled with carbon dioxide. Dosimetry for X- and gamma-radiation was by means of lithium fluoride thermoluminescent dosemeters calibrated against a Victoreen ionization chamber.


Somatic aberration induction in Tradescantia occidentalis by neutrons, x- and gamma-radiations. II. Biological results, r.b.e. and o.e.r. Biological results, including statistical features, are described for the irradiation of Tradescantia occidentalis with 250 kVp X-rays, cobalt-60 gamma-radiation and monoenergetic neutrons with energies between 0-08 and 15 MeV. The effect studied was that of the induction of pink sectors in the otherwise blue staminal hairs of the flowers at low doses of radiation. Statistical aspects of the results suggest that a fraction of the asynchronous cell population in the hairs is very sensitive to neutron radiation, but not necessarily to lower LET radiations. All the results were fitted by a least-squares method by polynomials of different degrees. Best fits to X- and gamma-ray data were provided by second-degree polynomials, and to the neutron data by either second- or third-degree polynomials. Limiting r.b.e. and o.e.r. values at low doses are derived. Some computed microdosimetric parameters are presented in comparison with the r.b.e. values. It is concluded that the effect studied is complex and may not provide a critical test of bio-physical theories of radiation effects.


Independence of the radiobiological oxygen constant, K, and the respiration rate of mammalian cells. The radiosensitivity, and the radiobiological oxygen-constant K for mouse Ehrlich ascites cells depend on the pre-irradiation growth conditions. Measurements of the oxygen-consumption rate of cells grown by different methods have been made to ascertain whether the differences in radiobiological properties could be associated with different rates of respiration. Inhibition of respiration by sodium amytal had no significant effect on either the radiosensitivity or the estimate of K.


The effects of x-irradiation on collagen metabolism in rat skin. Collagen metabolism in rat skin and the response to X-radiation are studied during three consecutive hair-cycles. Collagen biosynthesis and turnover, assessed by the hydroxylation of [U-14C]-proline, proceeds as a function of the periodic hair-growth phenomenon. A single X-radiation treatment to the skin does not affect the amino-acid composition of collagen, but reduces the proline hydroxylation capacity of the fibroblasts. The initial damage to the tissue, resulting in a reduced collagen concentration of the skin, particularly noticeable after several hair-cycles, is at least partially responsible for the delayed skin lesions after irradiation.


The metabolites of catecholamines in urine of patients irradiated therapeutically. The metabolites of catecholamines were determined in 24-hour urine samples of patients with genital carcinoma and treated by radio therapy. The patients were irradiated first with gamma-rays of radium and then with X-rays. The radium sources (80 mCi) were placed intracavitarily for 46 hours twice within 2 weeks. X-irradiation (800 R daily), applied 1 month after radium treatment, was delivered on four abdominal fields over 15 days. The quantities of excreted catecholamine metabolites during irradiation were compared with control values (obtained before irradiation) in the same patients. Gamma-irradiation provoked a significant increase in the excretion of 3-methoxy-4-hydroxy-mandelic acid, metadrenaline and normetadrenaline, as well as of homovanillic acid, whereas X-irradiation provoked only a significant increase in the excretion of free 3-methoxy-4-hydroxy-phenylglycol. The increased excretion might be explained: (1) in the case of radium application, by direct radiation-induced release of catecholamines from the peripheral symphathetic nerves; (2) in the case of X-irradiation, by putting in the motion the complex of early neuroendocrine reactions via irradiated adrenal medulla.


The rates of electron transfer from ClUra- and ClUraH-to p-nitroacetophenone. Pulse-radiolysis experiments demonstrate that both the radical ion CIUra-/- and the protonated radical CIUraH- Of chlorouracil will transfer electrons to p-nitroacetophenone (PNAP). The reaction of CIUraH- is complicated by unknown reaction with PNAP below pH 4-4. Rate constants measured for reactions (2) and (3) are kappa2=5-3 x 109 and kappa3=3-3 x 109 M-1 sec -1. See article. An independent determination of the rate-constant for dissociation of the radical ion (equation (1)) was also obtained (kappa1=0-7 x 105 sec -1). See article.


Induction by gamma-radiation of DNA synthesis in radicle cells of germinating seeds of Pisum sativum l. In radicle meristem cells of germinating seeds of the pea (Pisum sativum L) before the onset of replicative synthesis of DNA, irradiation with 2-3 krad of gamma-rays induced the incorporation of 3H-thymidine (3H-TdR). Maximum isotope incorporation was noted during the first 2 hours after irradiation. Higher doses of radiation suppressed 3H-TdR incorporation. It was not seen after gamma-irradiation of air-dried seeds, nor after fast-neutron irradiation. The replication inhibitors hydroxyurea and 5-aminouracil had no effect on the gamma-induced incorporation of 3H-TdR, Whereas caffeine and acriflavine inhibited it to some extent. It is suggested that the gamma-radiation-induced incorporation of 3H-TdR in meristem cells during the pre-replicative period may be connected with repair phenomena.


Changes in morphogenesis and developmental enzyme levels in Dictyostelium discoideum after gamma irradiation. Changes in the levels of specific activity of two enzymes believed to be involved in developmental regulation were observed after irradiating differentiating cells of Dictyostelium discoideum. Stimulation of the levels of specific activity of alkaline phosphatase occured after irradiation at the beginning of development and at the end of the aggregation period, but not after irradiation at the beginning of aggregation. A stimulation in UDP-glucose pyrophosphorylase specific activity was also observed, but to a lesser extent and only after irradiation at the end of aggregation. Dose-dependent delays in the appearance of peaks of specific activity were noted. The delay per unit dose was less when irradiation took place at the beginning of development as opposed to the beginning or end of the aggregation period. Radiation-induced delays in progression through visible developmental stages were almost identical to delays in enzyme appearance. Other radiation effects on morphogenesis included the induction of a migratory slug phase.


Surface markers and other characteristics of the lymphocyte in chronic lymphocytic leukemia. The vast majority of leukemic cells bear surface Ig. The class restriction of this surface Ig and other characteristics described for chronic lymphocytic leukemia (CLL) are considered as evidence for the monoclonality of the neoplastic proliferation. Different CLL cases with various expressions of surface Ig represent various degrees of block in the maturation of the lymphocytes. Through the use of other cell markers, it has been shown that most cases of CLL represent B cell neoplasia. Variations in the expression of the various B cell markers on the leukemic cells were observed. One case in which the leukemic cells were clearly T cell in character, and a single case of mixed B and T CLL, are described. The significance of the sheep red blood cell rosette-forming cells in CLL is discussed. Scanning electron microscopy (SEM) can be a useful adjunct to the identification of B- and T-derived lymphocytes. According to these criteria, leukemic cells are mostly of the B type, although variations in their surface architecture were noted. It is concluded that SEM alone cannot consistently distinguish between leukemic B and T cells. Electron microscope studies of mitogen-transformed CLL lymphocytes suggest that there is a residual normal B and T population of cells in addition to the predominant, abnormally reacting cells, which are mostly of B origin. Antigenic changes on the surface of CLL lymphocytes are suggestive of normal antigens present on young normal lymphocytes, rather than of the emergence of truly "tumor-specific" antigens. These antigens are independent of the T or B origin of the leukemia. CLL lymphocytes are shown to be defective in their ability to form caps with concanavalin A and anti-HL-A sera, and in their osmotic regulatory capacity.


New high-molecular decomposition products of natamycin (pimaricin) with intact lactone-ring. Mild acid treatment of natamycin (IV) results in biologically inactive aponatamycin (VI), an amphoteric substance with some natamycin-like chemical and physical properties. Aponatamycin contains one natamycin- and one natamycinolide-moiety. More drastic acid degradation of natamycin eliminates the aminosugar under formation of the dimer (VII) of  the hypothetical aglycone of natamycin, natamycinolide (V) as well as a non-ionic compound, the dimer of the 12-decarboxy-11-anhydro analogue of natamycinolide.


A syngeneic metastatic tumor model in mice: the natural immune response of the host and its manipulation. The cell-mediated immune response against a transplantable syngeneic metastatic solid tumor in mice was studied. The immune reactivity of spleen cells from tumor-bearing mice was found to vary during development of the tumor. For about a week after tumor transplantation, the spleen cells were able to protect recipient mice against challenge with tumor cells. Subsequently, the protective activity was replaced by an enhancing activity. Recipient mice that received tumor cells together with spleen cells from mice bearing tumors for about two or three weeks had a higher incidence of tumor takes and larger tumors than controls. This enhancement of tumor development was correlated with the size of the local tumor or metastases in the donors. The enhancing activity was found to be mediated by T lymphocytes and appeared to suppress the protective immune response of the recipients. We devised a system to strengthen the immune response of the host against the development of tumor metastases. In the tumor model used, removal of the local tumor after s.c. transplantation failed to prevent the development of lung metastases and death in most of the mice. However, syngeneic spleen cells which had been sensitized in vitro against tumor cells were found to serve as immunotherapeutic agents. Injection of such spleen cells into mice from which primary tumor implants had been removed surgically led to a markedly increased survival. Spleen cells from both normal and tumor-sensitized donors were effective, but splenocytes from mice bearing large tumors did not reduce metastatic development after sensitization in vitro. Thus, protection against the development of lethal metastases can be achieved with certain types of lymphocytes sensitized in vitro.


Selection and enrichment of antigen-specific T lymphocyte populations. Antigen-specific T lymphocytes bind in vitro to cells carrying alloantigens or to macrophages bearing antigen to which the lymphocytes were sensitized. This phenomenon is the basis of present procedures for the isolation and enrichment of antigen-specific T lymphocytes. The antigen-mediated adherence permits the removal of T cells with other specificities, which do not bind to the antigen-containing monolayer. The subsequent activation and multiplication of the antigen-bound cells further increases the proportion of the antigen-specific lymphocytes in culture. The selected cells, which are derived from a small fraction of the initial lymphocyte population, proliferate until they comprise up to 30% of the original total cell number. Another factor favoring enrichment is that unactivated cells do not survive well in culture whereas antigen-specific activated cells maintain their viability. Indeed, the selected cells can be maintained for a few weeks in culture. The activity of the selected lymphocytes is elevated toward the antigen used for enrichment, but lowered, or absent, toward other antigens. The progeny of the antigen-adherent lymphocytes are thus seen to be enriched for one antigen-specific subpopulation, and deprived of lymphocytes committed to other antigens. Cells obtained by this procedure should prove to be very useful for the functional analysis of T lymphocytes.


Comparison of the chemical, physical, and survival properties of normal and Z-variant alpha-1-antitrypsins. A procedure has been developed for the purification of Z-type alpha-1-antitrypsin (alpha-1-AT) which is rapid, gentle, and results in good yields. From 4 units (750 ml) of fresh human plasma, obtained from two individuals possessing the Pizz phenotype, 53 mg of pure Z-type alpha-1-AT was obtained. The preparation was homogeneous by the criteria of polyacrylamide gel electrophoresis, both in the presence and absence of sodium dodecyl sulfate, and by analytical ultracentrifugation. When compared to pure alpha-1-AT from plasma of individuals possessing the normal PiMM phenotype, the two proteins were indistinguishable with respect to amino acid composition, sedimentation coefficient (s20w of 3.33 for both M and Z), molecular weight (51,000 by sodium dodecyl sulfate gel electrophoresis and 47,000 by sedimentation equilibrium for both M and Z), and trypsin-combining ratio (0.91 for Z and 0.99 for M). The only difference which was observed between the variant forms of alpha-1-AT was in the carbohydrate composition. The Z-type alpha1-AT contains between 20 and 25% less carbohydrate than the M-type alpha-1-AT. Specifically, the Z-type alpha-1-AT is deficient in 1 glucosamine residue, 3 neutral sugar residues (1 mannose and 2 galactose), and 2 sialic acid residues. Although the Z-variant is deficient in sialic acid, its survival time in the serum of a rabbit was not significantly different from that of M-type alpha-1-AT.


Surface phenotype of nonadherent peritoneal cells effecting cell-mediated cytotoxicity in vitro for allogeneic and syngeneic murine sarcoma cells. The participation of B cells in cell-mediated cytotoxicity (CMC) for allogeneic and syngeneic murine sarcoma cells induced with 3-methylcholanthrene was investigated. Primed nonadherent peritoneal cells (NAPC) were treated with antisera against lymphocyte surface antigens and complement, and residual CMC activity was measured with the [3H]proline microassay. The antisera (anti-PC.1 and various anti-Ig sera) used for this purpose were highly reactive with B cells according to established serologic criteria. Elimination of cells that carried PC.1 or Ig on their surface did not diminish CMC of NAPC for allogeneic or syngeneic tumor cells. Exposure of cytotoxic NAPC to various anti-Ig antisera (without complement) before and during the CMC assay did not inhibit CMC either. We conclude that under these conditions CMC is not mediated by B cells, nor is it dependent on their presence. In addition, our findings do not implicate any of the usual classes and types of Ig covered by the Ig antisera used in this study as constituting specific receptors on the effector T cells of CMC.


Induction of cell-mediated immunity in the mouse: circumstantial evidence for highly immunogenic antigen in the regional lymph nodes following skin painting with contact sensitizing agents. This paper describes an investigation of why contact sensitizing agents cause strong cell-mediated immunity. Contact sensitivity was induced in mice by painting the skin with 4-ethoxymethylene-2-phenyloxazolone (oxazolone), and measured by the increase of ear thickness following challenge six days later. Reactivity was transferred by taking the regional lymph node cells from mice 18 h after immunization and injecting them into the footpads of recipients. This "18-h transfer" has several characteristics. As few as 2 X 10(4) cells were effective. The donor lymph node cells were best taken one to three days after immunization, were less effective on day 4 and virtually inactive by day 7. The recipients developed contact sensitivity when challenged on day 4, but lacked sensitivity when challenged on days 1 and 2 after transfer. The transferred cells were still active after treatment with anti-theta serum and complement. They also resisted 2,000 R in vitro, mitomycin, vinblastine, and inhibitors of protein synthesis such as emetine, cycloheximide and puromycin. The transfer was prevented by treatment with trypsin, freeze-thawing, and heating at 56 C. Plasma membranes were also immunogenic. The evidence suggests that the "18-h transfer" is a special type of active immunization, not due to ordinary free oxazolone, and that the agent is present within the lymph node in a free oxazolone, and that the agent is present within the lymph node in a specially immunogenic location or form.


Axonal transport of [3H]serotonin in an identified neuron of Aplysia californica. A population of characteristic ellipsoidal dense-core vesicles was identified in axons of the giant cerebral neuron of the mollusc Aplysia. We injected [3H]serotonin into the cell body of this identified serotonergic neuron in the isolated central nervous system in order to study the subcellular components associated with the neurotransmitter. Subcellular fractionation by differential centrifugation indicated that injected serotonin was rapidly taken up into particulate form. [3H]Serotonin appeared in the axons within 2 h after injection, and export continued at a constant rate of 6% of the total in the neuron/h thereafter. The dependence of the total amounts of [3H]serotonin which appeared in the axons in 6 h (export from the cell body) on the amounts injected was consistent with the idea that export is a saturable process, possibly depending on the capacity of somatic vesicles or of some unidentified carrier for serotonin. [3H]Serotonin moved into both major branches of the axon, where it was translocated rapidly. The transmitter, which was shown by autoradiography to be restricted to the axons of the injected cell, was distributed along axons in accumulations of radioactivity; in contrast, its precursor, [5-3H]hydroxytryptophan, moved slowly along axons in a smooth, declining curve, its kinetics consistent with diffusion. Quantitative electron microscope autoradiography revealed that the dense-core vesicles and the cytosol of axons fixed with glutaraldehyde were labeled with [3H]serotonin.


Human serum binding capacity and affinity for 25-hydroxyergocalciferol and 25-hydroxycholecalciferol. 25-Hydroxyvitamin D-binding capacity and affinity were studied in human cord, adult, and maternal sera, and in sera from women receiving oral contraceptives, by in vitro satuaration analyses employing dextran-coated charcoal to adsorb unbound sterol. 25-Hydroxyergocalciferol and 25-hydroxycholecalciferol were equipotent in their ability to displace 3H 25-hydroxycholecalciferol from human serum binding sites. At 0C, the apparent dissociation constant for the serum binding of 25-hydroxycholecalciferol was low (Kd=8x 10-10M). Cord and adult sera had a similar 25-hydroxycholecalciferol binding capacity (1.8 x 10-6M), but the binding capacity of maternal sera and the sera from women receiving oral contraceptives was significantly higher. At physiological serum concentrations of 25-hydroxyvitamin D (5 x 10-8M), only 2-3% of human serum 25-hydroxyvitamin D-bindig sites are occupied.


Studies on human antihemophilic factor. Evidence for a covalently linked subunit structure. When purified antihemophilic factor (Factor VIII) was rechromatographed on 4% agarose in 0.15 M NaCl or 1.0 M NaCl, a single protein peak, containing both procoagulant activity and von Willebrand factor activity, as defined by ristocetin-induced platelet aggregation, was eluted in the void volume. Purified Factor VIII immediately lost about 30% of its procoagulant activity when dissolved in 0.25 M CaCl2, and when rechromatographed on 4% agarose in 0.25 M CaCl2, the protein peak and von Willebrand factor activity remained coincident in the void volume; however, most of the remaining procoagulant activity was eluted after the void volume. The elution position of Factor VIII procoagulant activity from 4% agarose in 0.25 M CaCl2, and hence its apparent molecular weight, varied with the protein concentration applied to the column; at low protein concentrations it was eluted close to the inner volume. Yet on Sephadex G-200 in 0.25 M CaCl2, the protein and procoagulant activity were eluted together in the void volume. These observations suggested that the Factor VIII procoagulant activity was not eluting according to size or shape, but was adsorbing to some extent to the agarose. Isolated activity peak material from the 0.25 M CaCl2 columns contained protein and had a typical ultraviolet spectrum. Even at high concentrations, the protein contained no thrombin, Factors IX, X, or Xa activity, or detectable phospholipid. In addition to Factor VIII procoagulant activity, which could be inactivated by a human antibody to Factor VIII, the activity peak protein also contained von Willebrand factor activity. Like native Factor VIII and the void volume protein, the activity peak contained protein that did not enter a sodium dodecyl sulfate 5% polyacrylamide gel in the absence of reducing reagent. After reduction of disulfide bonds, several subunits ranging from 195,000 to 30,000 daltons were observed. These results indicate that the protein in the shifted Factor VIII procoagulant activity peak is large and that its anomalous elution pattern from 4% agarose in 0.25 M CaCl2 results from interaction with the agarose. The Factor VIII-like properties of the activity peak protein and its electrophoretic pattern on sodium dodecyl sulfate gels suggest that it is a species of Factor VIII modified by proteolytic cleavage. These results allow an interpretation that is different from the recently proposed "carrier protein-small active subunit" hypotheses for the structure-function relationships of the Factor VIII molecule.


Obstructive airway disease associated with heterozygous alpha-1-antitrypsin deficiency. Twenty-eight members of one family were investigated for the presence of pulmonary disease concurrent with variable (ZZ, MZ, or MS) degrees of alpha-1 antitrypsin deficiency. Three symptomatic siblings (2, ZZ; 1, MZ) had abnormal pulmonary scans and abnormal routine pulmonary function studies. Five asymptomatic heterozygotes, three of whom had never smoked and were the youngest studied, had distinctly abnormal closing volumes. Three of the five patients also had abnormal perfusion dynamics when scanning was done in the upright position. Pulmonary dysfunction occurs in asymptomatic heterozygotes but requires the use of sensitive techniques for its appreciation.


Low molecular weight immunoglobulins in Rana catesbeiana tadpoles. Rana catesbeiana tadpoles formed high and low m.w. antibodies in response to immunization with a bacteriophage. Although the neutralizing activity associated with the low m.w. immunoglobulins was relatively weak, the existence of antibodies in this class was clearly demonstrated by radioimmunoelectrophoresis. Moreover, two antigenically distinct variants of the low m.w. antibodies were detected. These were serologically indistinguishable from the two types of low m.w. immunoglobulin previously isolated from the serum of adult frogs of this species.


Function of 2-mercaptoethanol as a macrophage substitute in the primary immune response in vitro. The mechanism by which 2-ME acts as a macrophage-substitute for the induction of a primary PFC response to SRC in vitro was studied in macrophage-depleted mouse spleen cell cultures. 2-ME could replace macrophages only in FCS-supplemented cultures. Evidence is presented that the function of 2-ME is independent of residual macrophages. Neither normal nor macrophage-depleted spleen cell cultures from congenitally athymic nude mice supplemented with 2-ME, with or without FCS, could give rise to a primary in vitro anti-SRC immune response. 2-ME, at an optimal concentration of 10(-5) M, induced DNA synthesis in normal and macrophage-depleted spleen cells in both FCS-containing and serum-free cultures. The peak response occurred on day 3. The stimulation was accompanied by a polyclonal B cell activation to antibody secretion which was much more pronounced in FCS-containing than in serum-free cultures. Spleen cells from nude mice showed a weaker DNA stimulation than did cells from normal mice in FCS-containing cultures, and nearly no response under serum-free conditions. T cells obtained by a nylon column adherence method from normal mouse spleen cells showed good DNA synthetic responses in FCS-containing, but no response in serum-free cultures. These results show that 2-ME has weak mitogenic activity for B cells, and in combination with FCS, strong mitogenic activity for T cells. Since the macrophage provides stimulation to the T cell in the primary anti-SRC PFC response in vitro, these results suggest that the direct mitogenic activity of 2-ME with FCS on T cells provides the functional substitution for macrophages.


Immunogen structure and cell selection. I. Cellular requirements for the in vitro response to moderately haptenated forms of the solid-phase immunogen DNP-O-Bio-Gel-P. The cellular requirements for the in vitro response to DNP-O-Bio-Gel-P were determined. Results of these experiments indicate that: 1) splenic adherent cells are required for generation of anti-hapten PFC to the solid phase immunogen; and 2) cultured nude-mouse spleen cells have far less capacity to respond to the immunogen than do cells from wild type mice. These experiments suggest that both T and B cells are required for responsiveness to the moderately haptenated form of DNP-O-Bio-Gel-P.


Two functionally distinct anti-tumor effector cells isolated from primary murine sarcoma virus-induced tumors. The ability of cells from primary MSV-induced tumors to function as effector cells in vitro was evaluated. Host cells were isolated by enzymatic disaggregation of the tumor and fractionated by sedimentation velocity at unit gravity on a Ficoll gradient. Characterization of these cells indicated that 30 to 40 % were T lymphocytes, about 50% were macrophages and less than 5% were B lymphocytes. Two different functional activities were mediated by these cells: cytolysis, as measured by the CRA, and inhibition of proliferation, as measured by the GIA. The effector cells in the CRA were T cells with sedimentation velocities of 3.5 to 4.0 mm/hr, whereas those cells which mediated the GIA were presumably macrophages and displayed a heterogeneity in size two peak sedimentation velocities, one at 4.0 mm/hr and another at 6.0 mm/hr. Activity by the effector cells in the CRA was antigen specific in contrast to the activity in the GIA which was directed against cells which did not carry detectable cross-reacting antigens.


B lymphocytes in the guinea pig thymus are specifically localized in the central area. Since the central area is an integral part of the guinea pig thymus, the cells in this area were compared with those in the thymic cortex and medulla in cryostat-sections by using methods for demonstration of E-, EA-, EAC-adherence and surface membrane immunoglobulins. In the extra cortical central area (ECCA) 15 to 25% of the lymphocytes showed EAC-adherence and 5 to 10% appeared to bear surface membrane immunoglobulins (SIg). In the lymph sinuses up to 70% of the lymphocytes were EAC- and SIg-positive. A small amount of EAC-adhering cells was present in the medulla of the central area. Cortical lymphocytes were EAC- and SIg-negative. From these results we conclude that in the guinea pig thymus B lymphocytes are specifically localized in the central area.


Requirements for induction of T cell tolerance to DNFB: efficiency of membrane-associated DNFB. Tolerance to contact sensitization with DNFB, a T cell-dependent phenomenon, was induced in mice by preparations of DNFB coupled to mouse RBC or spleen cells. Such tolerance is dose related, wanes with time, and can be transferred to normal animals with lymphoid cells (presumably containing suppressors). Tolerance to DNFB-RBC can be produced by whole DNFB-RBC, by ghosts of these cells, by sonicates of the ghosts, and by detergent-treated DNFB-RBC ghosts. Tolerance cannot be produced by larger amounts of DNFB-RBC components not associated with membrane. The ability of various DNP compounds to stimulate DNA synthesis in DNFB-sensitized cells also correlates with their ability to bind to protein components; i.e., DNFB is a far more efficient stimulator than DNBSO, whereas DNPlysine does not stimulate at all. Thus, the ability to sensitize or to tolerize with DNFB congeners is related to their ability to couple to proteins. It appears that the active induction of T cell tolerance requires that tolerogen be coupled to cell membranes. Since both T cell sensitization and tolerance to DNFB are best produced by DNFB-membrane, the actual occurrence of one state or the other must depend on the molecular method of "presentation" of DNFB-membrane.


Rat T lymphocyte-specific antigens and their cross-reactivity with mouse T cells. Anti-rat T lymphocyte serum (ATLS)2 was prepared by immunizing rabbits with purified T cells from rat mesenteric nodes and absorbed with rat red cells and syngeneic sarcoma cells. The specificity of ATLS for rat T cells was confirmed by the following reasons: a) ATLS was not toxic for bone marrow cells but lysed most of the thymocytes and a number of spleen and lymph node cells, which were inversely correlated to the percentage of cells with B cell characteristics in respective organs; b) anatomical localization of ATLS-reactive cells in lymphoid organs coincided to the thymus-dependent areas, i.e. the paracortex of lymph node and the periarteriolar region of spleen; c) spleen cells treated with ATLS and complement failed to respond to phytohemagglutinin but normally responded to bacterial lipopolysaccharide; d) those cells treated with ATLS and complement could not induce a graft-vs-host reaction in F1 hosts, whereas the same treatment did not affect direct plaque-forming cells. All of these data confirm the specificity of ATLS and indicate that ATLS recognizes rat T lymphocyte-specific antigens (RTLA). Absorption studies showed that RTLA were present in higher concentration on medullary thymocytes and peripheral T cells than on cortical thymocytes, but absent from bone marrow, liver, and brain tissues. When the cross-reactivity of RTLA with mouse T cells was studied by C-dependent cytotoxicity and immunofluorescence, it was found that mouse T cells shared at least one determinant of RTLA with rat T cells, and that distribution pattern of the cross-reacting antigens in mouse lymphoid tissues was essentially the same as that of RTLA in rat lymphoid organs.


Isolation of a murine leukemia FC receptor by selective release induced by surface redistribution. A protein which binds to the Fc region of IgG has been isolated from the murine leukemia L1210. The isolation technique involves surface cross-linking of the cells's Fc receptors with the use of aggregated human IgG and anti-human IgG. This results in the redistribution (patch formation and capping) of the cells's Fc receptors. Lactoperoxidase-catalyzed radioiodination of the cells before complex binding indicates that Fc receptor redistribution results in the selective release of surface proteins. SDS-PAGE analyses of the supernatants from cells thus treated reveals a major peak corresponding to a molecular weight of 45,000 daltons. This protein has been purified from the cell supernatants by immunoprecipitation and chromatography of the percipitates on Sephadex G-200 under dissociating conditions. After separation from the immune complex this protein can be bound to heat-aggregated IgG, but not aggregated F(ab')2 fragments. The 45,000 dalton protein appears to be the Fc receptor which has been released from the cell surface in association with the complex.


T lymphocyte-enriched murine peritoneal exudate cells. III. Inhibition of antigen-induced T lymphocyte Proliferation with anti-Ia antisera. The recent development of a reliable murine T lymphocyte proliferation assay has facilitated the study of T lymphocyte function in vitro. In this paper, the effect of anti-histocompatibility antisera on the proliferative response was investigated. The continuous presence of anti-Ia antisera in the cultures was found to inhibit the responses to the antigens poly (Glu58 Lys38 Tyr4) [GLT], poly (Tyr, Glu) ploy D,L Ala-poly Lys [(T,G)-A--L], poly (Phe, Glu)-poly D,L Ala-poly Lys [(phi, G)-A--L], lactate dehydrogenase H4, staphylococcal nuclease, and the IgA myeloma protein, TEPC 15. The T lymphocyte proliferative responses to all of these antigens have previously been shown to be under the genetic control of major histocompatibility-linked immune response genes. The anti-Ia antisera were also capable of inhibiting proliferative responses to antigens such as PPD, to which all strains respond. In contrast, antisera directed solely against H-2K or H-2D antigens did not give significant inhibition. Anti-Ia antisera capable of reacting with antigens coded for by genetically defined subregions of the I locus were capable of completely inhibiting the proliferative response. In the two cases studied, GLT and (T,G)-A--L, an Ir gene controlling the T lymphocyte proliferative response to the antigen had been previously mapped to the same subregion as that which coded for the Ia antigens recognized by the blocking antisera. Finally, in F1 hybrids between responder and nonresponder strains, the anti-Ia antisera showed haplotype-specific inhibition. That is, anti-Ia antisera directed against the responder haplotype could completely block the antigen response controlled by Ir genes of that haplotype; anti-Ia antisera directed against Ia antigens of the nonresponder haplotype gave only partial or no inhibition. Since this selective inhibition was reciprocal depending on which antigen was used, it suggested that the mechanism of anti-Ia antisera inhibition was not cell killing or a nonspecific turning off of the cell but rather a blockade of antigen stimulation at the cell surface. Furthermore, the selective inhibition demonstrates a phenotypic linkage between Ir gene products and Ia antigens at the cell surface. These results, coupled with the known genetic linkage of Ir genes and the genes coding for Ia antigens, suggest that Ia antigens are determinants on Ir gene products.


Cellular specificity of plasmacytoma-induced immunosuppression. The pattern of immunodeficiency in plasmacytoma-bearing mice appears to be unique. Mice bearing these tumors exhibit a severe impairment in their ability to mount a primary immune response to thymus-dependent and -independent antigens. However, cell-mediated immune functions in these mice apparently remain intact. Thus, when T cell activity of lymph node cells from plasmacytoma-bearing mice was tested in vivo by sensitization with dinitrofluorobenzene and in vitro by responsiveness to phytohemmagglutinin, allogeneic cells, and dinitrobenzene sulfonate, cell-mediated immunity was found to be normal.


T cell requirement for experimental allergic encephalomyelitis induction in the rat. The question of whether a cell-mediated or a humoral mechanism initiates EAE in rats sensitized with BP-CFA was investigated. The requirement of T cells for EAE induction was manifested when Tx, irradiated rats were reconstituted with normal lymphoid cells treated with ATS and then injected with BP-CFA. Neither EAE nor antibody was produced, indicating the T cell dependency of BP specific antibody production. More precise information regarding the role of the T cell in the production of EAE was obtained by means of passive transfer of EAE with sensitized lymphocytes. Thus, transfer of lymphoid cells from rats previously sensitized to BP-CFA into Tx, irradiated rats elicited EAE and antibodies to BP. However, no EAE followed when the transferred cells were first depleted of T cells by treatment with ATS. Nevertheless, ATP pretreatment did not depress the levels of antibody to BP produced in the transfer recipients. The latter finding indicates that the cells from animals sensitized 9 days previously were already committed to the production of antibodies to BP. Therefore, a) T cells are absolutely necessary for induction of EAE and b) antibody detected by antigen-binding is not responsible for the pathogenesis of this disease.


Recruitment of effector lymphocytes by initiator lymphocytes. Circulating lymphocytes are trapped in the reacting lymph node. In previous studies, we have shown that initiator T lymphocytes (ITL) sensitized in vitro recruit effector T lymphocytes in vivo. When ITL were sensitized against fibroblast antigens in vitro and injected into footpads of syngeneic recipients, they induced enlargement of the draining popliteal lymph node (PLN) and the development there of specific effector lymphocytes of recipient origin. To study the basis of this lymph node response in recruitment, we injected 51Cr-labeled spleen cells i.v. into recipients of sensitized ITL and found that the labeled circulating lymphocytes were trapped in the reacting PLN. The trapping depended on surface properties of the labeled circulating lymphocytes, as revealed by various enzymatic treatments. The trapping process was radiosensitive, both on the part of the trapped lymphocytes and the lymph node-trapping mechanism. Thus, sensitized ITL injected into the hind footpads migrate to the PLN and induce the trapping of circulating recruitable lymphocytes, which either differentiate into or regulate the differentiation of effector T lymphocytes.


A temporal dissociation of energy liberation and high energy phosphate splitting during shortening in frog skeletal muscles. Measurements of the time course of high energy phosphate splitting and energy liberation were performed on rapidly shortening Rana pipiens skeletal muscles. In muscles contracting 30 times against small loads (less the 0.02P), the ratio of explained heat + work (H + W) (calculated from the measured high energy phosphate splitting) to observed H + W (from myothermal and mechanical measurements) was 0.68 +/- 0.08 and is in agreement with results obtained in isometric tetani of R. pipiens skeletal muscle. In lightly afterloaded muscles which were tetanized for 0.6a and whose metabolism was arrested at 3.0 s after the beginning of stimulation, a similar ratio of explained H + W to observed H + W was obtained. However, in identical contractions in which metabolism was arrested at 0.5-0.75 s after the beginning of stimulation, the ratio of explained H + W to observed H + W declined significantly to values  ranging from 0.15 to 0.40. These results suggest that rapid shortening at the beginning of contraction induces a delay between energy production and measurable high energy phosphate splitting. This interpretation was tested and confirmed in experiments in which one muscle of a pair contracted isometrically while the other contracted against a small afterload. The afterload and stimulus pattern were arranged so that at the time metabolism was arrested, 0.5 s after the beginning of stimulation, the total energy production by both muscles was the same. Chemical analysis revealed that the isotonically contracting muscle spilt only 25% as much high energy phosphate as did the isometrically contracting muscle.


A reexamination of the thermoelastic effect in active striated muscle. Isolated Rana pipiens sartorius muscles at 0degreeC were stimulated via their nerves and small stretches or releases applied during the plateau of the isometric tetanus at lo. Extra heat above the isometric maintenance heat was produced during the drop in tension caused by release and, for very small releases (delta less than or equal to 0.5% lo), was completely reabsorbed during tension recovery. The extra heat was always directly proportional to the tension change. Heat absorption proportional to the tension change was also observed during the increase in tension produced by small stretches in the range 0.5% lo less than or equal to deltal less than or equal to 3.0% lo. The mean heat:tension ratio R in seven experiments was -0.0084, which is within the range of values reported previously by Woledge. In addition, it was found that during tension recovery after small releases of 1.0% lo less than or equal to deltal less than or equal to 3.0% lo the "contractile" component seems able to shorten about 1% lo without producing shortening heat.


Malignant spread of haemangioblastoma: report on two cases. Two cases are described in which, after successful removal of a cerebellar haemangioblastoma followed by several years of freedom from symptoms, there developed a progressive spinal cord compression, leading to death. At necropsy the spinal cords in both cases and the brainstem in one case, were irregularly plastered with haemangioblastoma. Although there was no doubt that malignant spread had occurred from one or more primary tumours, the histology of the tumour tissue was in no way different from that of conventional haemangioblastoma.


Microtubules in synapses of the retina.

Microtubules in synapses of the retina. Using a new method, microtubules can be seen running up to, and lying in close relationship with, the synaptic ribbons in the outer and inner plexiform layers of the frog retina. In the inner plexiform layer microtubules can be seen running up to the terminal membrane in the non-ribbon synapses. Unlike non-ribbon C.N.S. synapses (frog and rat) processed by the same method. There is no clear association between synaptic vesicles and microtubules in the approach regions.


Electrical properties of frog motoneurons in the in situ spinal cord. Electrical properties of the spinal motoneurons of Rana temporaria and R. esculenta were investigated in the in situ spinal cord at 20-22 degrees C by means of intracellular recording and current injection. Input resistance values depended on the method of measurement in a given cell but were generally inversely related to axon conduction velocity. The membrane-potential response to a subthreshold current pulse was composed of at least two exponentials with mean time constants of 2.5 and 20 ms. The membrance potential reached by the peak of a spike depended on the mode of spike initiation and membrane potential. Preceding a suprathreshold depolarization by a hyperpolarizing pulse could delay and eliminate spike initiation, similar to effects reported in certain invertebrate neurons. Antidromic invasion frequently failed in motoneurons of normal resting potential. Antidromic spike components (m,IS, SD) were similar to those of cat motoneurons. The delayed depolarization and the long afterhyperpolarization following an antidromic spike had many properties in common with the analogous afterpotentials of cat motoneurons. The reversal potential of the short afterhyperpolarization occurring immediately after the spike varied with resting potential and could not be used to determine potassium equilibrium potential. Sustained rhythmic firing could be evoked by continuous synaptic drive or long pulses of injected current. The plot of firing rate versus current strength had a substantial linear region. Both steady firing and adaptation properties varied markedly with motoneuron input resistance.


Evidence for electrotonic coupling between frog motoneurons in the in situ spinal cord. A recurrent EPSP was observed on antidromic stimulation of motoneurons in the in situ spinal cord of Rana temporaria and R. esculenta at 20-22C. The EPSP was finely graded and not refractory following full or partial antidromic spike components in a given neuron. The EPSP amplitude varied in parallel with the antidromic field potential under different conditions, suggesting transmission of the EPSP to the recorded motoneuron depended on invasion of the somadendritic membrane or neighboring motoneurons by the antidromic spike. The latency of the EPSP with respect to antidromic invasion of the local motoneuron pool was too short for the EPSP to be mediated by chemical transmission. It was concluded the EPSP was electrically transmitted between the somadendritic membranes of the motoneurons. Under certain conditions, the EPSP magnitude could be made to vary with membrane potential in a direction opposite to that expected from a chemical EPSP. Dendritic spikes were sometimes associated with the EPSP.


Direct effect of electrical stimulation on peripheral nerve evoked activity: implications in pain relief. Experiments were performed with a peripheral neurostimulator, used clinically for pain relief, on isolated cat cutaneous peripheral nerve to determine the effect of electrical stimulation on components of the compound action potential. The results show that neurostimulation alters the conduction velocity and the amplitude of both the A-alpha and beta and the A-delta waves with the more slowly-conducting A-delta component showing the greatest changes. This direct alteration of peripheral nerve activity distal to the first synapse in the spinal cord might contribute to the mechanism of pain relief.