Coordination of the head and eyes in pursuit of predictable and random target motion. Subjects were required to use their head and eyes in pursuit of visual targets which moved randomly or sinusoidally in the horizontal plane. All subjects disliked moving their heads to pursue the random motion, apparently because the motion broke fixation which resulted in a predominance of the vestibulo-ocular compensatory reflex over the smooth pursuit reflex. As a consequence gaze (head plus eye movement) was at times in the opposite direction to the motion of the target. In steady state pursuit of sinusoidal targets, eye movement consisted of a combination of pursuit and vestibulo-ocular reflex eye movements. At frequencies below 0.8 HZ, the vestibular reflex was used at times of minimum target velocity to stabilize fixation whereas during maximum target velocity the head movement was slowed and the smooth pursuit reflex predominated. At 1 HZ and over, there was a failure to suppress the compensatory vestibulo-ocular reflex; however, the saccades of vestibular nystagmus were used to "catch up" the target. There was a preference not to use the head in predictable pursuit.


[Emergency colectomy in ulcerative colitis (author's transl)]. During the last 12 years 7 patients with ulcerative colitis were treated because of an acute complication consisting in 2 cases of colonic perforation, 3 cases of toxic megacolon (two of them perforated) and 2 cases of massive enterorragia. In all patients an emergency total colectomy was performed resulting a mortality of 3 cases, two of which had required the removal in a second time of the rectal stump. In 3 or 4 recovered patients we performed in a period of 7-15 months an ileo-proctostomy once confirmed the healing of the ulcerative lesions in the residual rectum. The problems concerning the indications and surgical procedures involved in such cases are discussed on the base of the personal experience, with particular consideration on advantages and limits of preserving a rectal stump. The authors agree on an eclectic behavior considering each case on the base of pathological findings, the age and the general and psychic conditions of the single patient.


Treatment of advanced bronchogenic carcinoma with adriamycin, 5-fluorouracil and methotrexate. The combination of adriamycin, 5-fluorouracil and methotrexate with folinic acid reversal was used to treat patients with advanced bronchogenic carcinoma. Twenty-one of 30 patients showed an objective response. This combination appears to produce useful remissions in patients with non-squamous lung cancer.


The effect of androstanes on granulopoiesis in vitro and in vivo. Bone marrow culture in a patient with aplastic anaemia responding to anabolic steroid (methandienone) therapy, showed an unusually high degree of growth in unstimulated cultures. Growth in unstimulated cultures is due to factors with colony stimulating activity (CSA) released by monocyte macrophages in the bone marrow sample. These cells, which can be identified by staining for non-specific esterase activity, were not increased in this patient's marrow, implying either increased production of colony stimulating factors or increased sensitivity to these factors. Addition of methandienone or testosterone to feeder-layers containing normal peripheral blood leucocytes increased their stimulatory activity. Addition of these drugs to feeder-layers of CSF-containing conditioned medium did not have this effect, implying that androstanes cause increased production of colony stimulating factors rather than increased sensitivity to them. Lack of response to androstane therapy may be related to lack of response by bone marrow monocyte macrophages or to the inability of granulopoietic cells to respond to the increased CSF production induced by androstane therapy. A means of predicting lack of response is proposed.


Control of light-activated phosphorylation in frog photoreceptor membranes. In this paper, we examine some factors which regulate the efficiency of light in activating rhodopsin phosphorylation. We have measured phosphate incorporation after illumination in suspensions of bullfrog rod outer segments incubated with [gamma-32P]ATP. We observed that delaying ATP addition after illumination causes maximum phosphate incorporation to decrease 80% within 2 h. This decay occurs in urea-treated, extracted rod outer segment membranes. The decay of the light effect is not influenced by regeneration of opsin to rhodopsin or the presence of long-lived photoproducts. However, regeneration of opsin increases the amount of phosphorylation initiated by a second exposure to light. Further phosphorylation can also occur after phosphate groups have been removed from the membranes by dephosphorylation. Finally, we have confirmed our earlier observation that small amounts of light (bleaching less than 5% of the rhodopsin present) are more effective, by tenfold, in initiating phosphorylation than are larger amounts.


Oxygen consumption by frog skin and its isolated epithelial layers as a function of their sodium-transporting activity. The metabolic cost (in terms of oxygen consumption) of transcellular sodium transport was assessed on ventral frog skin and its isolated epithelial layers, by measuring the decrease in oxygen consumption by the tissue upon transient withdrawal of sodium from the outside solution. The same number of sodium ions was transported per molecule oxygen consumed whole skin (17.4 +/- 2.3) and its isolated epithelium (17.3 +/- 2.4). The metabolic cost of sodium transport could not be estimated properly when this process was blocked by amiloride or ouabain, as these drugs were found to bring about an increase in oxygen consumpton by the tissue when no sodium was available for transport.


Magnetic field effects on radical pair intermediates in bacterial photosynthesis. We have investigated the effects of magnetic fields on the formation and decay of excited states in the photochemical reaction centers of Rhodopseudomonae sphaeroides. In chemically reduced reaction centers, a magnetic field decreases the fraction of the transient state PF that decays by way of the bacteriochlorophyll triplet state PR. At room temperature, a 2-kG field decreases the quantum yield of Pr by about 40%. In carotenoid-containing reaction centers, the yield of the carotenoid triplet state which forms via PR is reduced similarly. The effect of the field depends monotonically on field-strength, saturating at about 1 kG. The effect decreases at lower temperatures, when the yield of PR is higher. Magnetic fields do not significantly affect the formation of the triplet state of bacteriochlorophyll in vitro, the photooxidation of P870 in reaction centers at moderate redox potential, or the decay kinetics of states PF and PR. The effect of magnetic fields support in view that state PF is a radical pair which is born in a singlet state but undergoes a rapid transformation into a mixture of singlet and triplet states. A simple kinetic model can account for the effects of the field and relate them to the temperature dependence of the yield of PR.


[Mechanism of the formation of single-stranded DNA breaks under the effect of prednisolone on rat liver thymocytes]. Rat liver thymocytes were incubated in vitro with prednisolon, and molecular weight of single-stranded DNA was estimated in alkaline lysates of nuclei by viscosity methods. The number of single-stranded breaks was estimated, which originate in DNA after 3 hours of incubation with prednisolon at a concentration of 3-10(-9) M--3-10(-4) M. Maximal hormone effect was produced within 30 min., and it remained constant for the rest 3 hours of the incubation. Cycloheximide prevented hormone-induced DNA depolymerization, and actinomycin D only slightly inhibited the formation of single-stranded DNA breaks, originated after 30 min. incubation of thymocytes with prednisolon, were repaired after their washing and reincubation in the fresh medium for 1 hour at 37 degrees C. The washing of thymocytes and their reincubation for 3 hours resulted in the decrease of the molecular weight of single-stranded DNA. Possible mechanism of the formation of single-stranded DNA breaks in prednisolon-induced thymocytes is discussed.


In vitro synthesis of alpha-1-antitrypsin in long term monolayer human liver cell cultures. Explants for long term monolayer liver cell cultures were taken from individuals having PiM or PiZ phenotypes. Radioactive labelling, using I125 and an original method, permitted the quantitative measurement of AAT in vitro and more important, allowed Pi phenotype determination.


Interaction of colony stimulating factor (CSF) from different sources with sepharose-concanavalin A. Preparations from various sources (human urines, mouse placenta and pregnant uterus extract, and fibroblast conditioned medium) were examined comparatively with affinity chromatography on Sepharose-Concanavalin A for the presence of Colony Stimulating Factor (CSF). The preparations from all sources constantly revealed two protein peaks: the proteins of the first peak, without Concanavalin A affinity, contained CSF in the material from pregnant uterus and fibroblast conditioned medium, while human urine preparation was devoid of CSF: the inhibitor of CFUc growth, detected and titrated in this peak, probably does not explain the lack of CFUc growth. The proteins from the second peak, specifically eluted with alpha-methyl D-glucopyranoside revealed CSF in all the examined preparations.


[Analgesia following electrostimulation of midbrain nuclei of rats with a pain syndrome of spinal origin]. The effect of electrostimulation of the mesencephalic grey matter and of the dorsal nucleus raphe on physiological pain produced by nociceptive stimulation (compression of the tail or the skin on the limb by a clamp) and on pathological pain (the pain syndrome of spinal origin) were studied in experiments on albino rats. Pathological pain was induced by creating a generator of pathologically enhanced excitation in the dorsal horn of the spinal cord by local disturbance of the inhibitory mechanisms with the aid of tetanus toxin. It was shown that electrostimulation of the indicated areas abolished both physiological and pathological pain. A conclusion was drawn that analgesia produced by electrostimulation of certain structure of the brain was connected not only with augmentation of the descending inhibition in the spinal cord as in the case of physiological pain caused by peripheral nociceptive stimulation (as shown by several authors), but also with the block of excitation at the supraspinal level. This mechanism should play a decisive role in analgesia realization in the pain syndrome of central origin, both under experimental and natural conditions.


[Changes in the concentration of catecholamines in the organs of animals with experimental myocardial infarcts under the influence of malaben]. The content of adrenaline and noradrenaline in the tissues of the heart, adrenal glands, spleen and brain of rats was studied in experimental myocardial infarction. A significant decrease in the catecholamine levels was revealed in the tissues. Malaben promoted normalization of the catecholamine tissue content in myocardial infarction. It is suggested that the said effect of malaben is due to its antihistaminic properties.


[Effect of cortisone-resistant thymocytes on endogenous colony formation in inbred mice]. Endogenous colonies in the spleen of sublethally irradiated (CBA X C57BL/6) E1 hybrid mice were recorded after the injection of thymocytes and the lymph node cells from the hydrocortisone-treated and intact CBA mice. Cortisone-resistant thymocytes failed to suppress the endogenous colony formation, whereas the lymph node cells produced a distinct suppressive effect on the endogenous colonies. After the injection of cortisone-resistant thymocytes the number of colonies in the spleen of individual recipients was double that in control irradiated hybrids.


PHA and endotoxin stimulation of human lymphocytes separated by albumin density gradient centrifugation. Venous blood from eight healthy subjects was divided into four fractions on a discontinuous albumin density gradient. The percentage recovery of lymphocytes was 82.3%; the purity of the lymphocyte fractions was 83.6%. The lymphocytes were cultured with PHA and Endotoxin, and the samples were analysed after 24, 48, and 72 hours. After PHA stimulation immunoblasts appeared up to 59.3% in the cultures from the 19-21% albumin fraction. After Endotoxin stimulation the maximum (75.8%) was reached in the heavy (25-27% albumin) fraction. Thus, it is concluded that the lymphocytes which can be stimulated with both the mitogens have different densities, the PHA-stimulable T lymphocytes being ligther than the Endotoxin-stimulable B lymphocytes. It is also concluded that as a mitogen Endotoxin is equal to PHA.


Comparison of the effects of caffeine and a 2-alkyl-1,2,3-benzotriazinium iodide on frog rectus abdominis. 1 The mode of action of 2-n-propyl-4-p-tolylamino-1,2,3-benzotriazinium iodide (TnPBI), which induced contractures of frog rectus abdominis muscle, was investigated. 2 TnPBI caused contractures of frog rectus abdominis when the muscle had been depolarized with potassium chloride. 3 Experiments with TnPBI and caffeine in calcium-free Ringer suggested that both compounds produce contractures by releasing intracellular bound calcium. 4 It is suggested that at least two calcium stores are involved, one of which is sensitive to caffeine while both are sensitive to TnPBI.


Bleeding in renal failure: is von Willebrand factor implicated? Normal or increased concentrations of factor VIII procoagulant activity (VIIIC), factor VIII-related antigen (VIIIRA), and factor VIII-von Willebrand activity (VIIIVWF) were found in the predialysis plasma of 10 out of 11 patients with chronic renal failure (CRF). All patients had a bleeding time longer than 15 minutes and platelet retention to glass beads lower than 40%. The only patient who had reduced concentrations of all three factor VIII complex components was subsequently shown to have von Willebrand's disease. In four patients with CRF, very low platelet retention, and slightly prolonged bleeding time none of the three factor VIII COMPLEX COMPONENTS WERE SELECTIVely modified in predialysis samples. These findings suggest that the bleeding tendency common in CRF is not necessarily linked to defective plasma factor VIII-related activities.


The spontaneous firing patterns of forebrain neurons. III. Prevention of induced asymmetries in caudate neuronal firing rates by unilateral thalamic lesions. Unilateral lesions interrupting striatal outputs and inputs (MFB lesions) produce a marked slowing of neuronal firing in the caudate nucleus contralateral to the side of the lesions without affecting neuronal firing in the ipsilateral caudate nucleus. Although the MFB lesion also interrupts the nigrostriatal pathway and depletes the ipsilateral striatum of dipamine and its associated enzymes, the slowing of unit firing rates is apparently due to interruption of striatal outputs rather than inputs. Unilateral thalamic lesions palced ipsilateral to MFB lesions in iether the ventral anterior-ventrolateral nuclei (VA-VL) or in the center median-parafascicular nuclei (CM-PF) prevent the MFB lesion-induced asymmetry in caudate neuronal firing rates. These thalamic lesions do not, however, restore the striatal dopamine content depleted by the MFB lesion. Unilateral CM-PF lesions in otherwise intact cats do not alter caudate unit firing rates nor do they affect striatal dopamine. VA-VL lesions in otherwise intact cats produce a bilateral slowing in the spontaneous firing of neurons in the caudate nuclei, again, whithout altering caudate dopamine concentrations. These results provide further evidence that caudate dopamine concentration per se does not appear to be a potent variable in controlling the spontaneous firing rates of striatal neurons.


Age related changes in the effect of electroconvulsive shock (ECS) on the in vivo hydroxylation of tyrosine and tryptophan in rat brain. A single electroconvulsive shock (ECS) was applied to 40 young (9 months) and 40 old (24 months) male rats. The effect on brain catecholamine synthesis was determined at intervals after ECS by measuring the accumulation of dihydroxyphenylalamine (DOPA) and 5-hydroxytryptophan (5-HTP) in rats pretreated with a central decarboxylase inhibitor (M-hydroxybenzyl hydrazine, NSD 1015). This accumulation permitted an estimation of the in vivo rates of hydroxylation of tyrosine and tryptophan. Control brain DOPA and 5-HTP concentrations were both lower in the older animals. Following ECS there was an increase in brain DOPA concentration (maximal at 60 min after ECS) in both young and old rats, but the increase was much smaller in the older animals. There were no changes in brain 5-HTP at any time after ECS, in either age group. It appears that aging selectively affects the response of the brain dopaminergic neurotransmitter system to stress, and we suggest that this may be a factor in the decreased resistance to stress in older subjects.


Origin of glutamate-decarboxylase (GAD)-containing cells in discrete hypothalamic nuclei. Glutamate decarboxylase activity (GAD) was assayed in discrete hypothalamic nuclei in the rat following lesions of the major afferent pathways from hippocampus, amygdala, midbrain, septum, thalamus and globus pallidus. None of these lesions led to a marked decrease in GAD activity in selected nuclei. After total deafferentation of the medial-basal hypothalamus GAD remained unchanged in the median eminence but fell markedly in the ventromedial and arcuate nuclei. In these two nuclei a decrease of GAD still occurred following partial deafferentation from lateral and posterior hypothalamus, but not from anterior and preoptic areas. These results indicated that most GAD-containing cells have their origin inside of the hypothalamus. In this region GABAergic neurons are likely to be short interneurons providing intrahypothalamic connections. Such connections are suggested from lateral and posterior hypothalamus onto the medial basal nuclei.


A measure of extracellular unit responses to repeated stimulation applied to observations of the time course of olfactory responses. A technique for measurement of small samples of unit behavior during regularly repeated stimulation is described. A correlation-based measure was computed over two successive stimulus presentation by summing the products of spike counts for corrsponding time bins and normalizing to the number of bins and the number of spike counts during the two stimulus presentations. This measure was combined with the mean frequency of spike occurrence during the stimulus presentation to give a characterization of neuronal activity sensitive to changes in both strength of temporal patterns and mean frequency. Examples are given of olfactory responsive neurons with comparison of measurement techniques. The time course of the response to odor of neurons recorded under urethane anesthesia from the olfactory bulb and the lateral hypothalamus was studied. By both simple mean frequency measures and the measures proposed in this study, the neurons recorded from the lateral hypothalamus underwent more rapid temporal decrements in the odor response.


Peripheral B- and T-lymphocyte counts in patients with sarcoma and breast carcinoma. Peripheral B- and T-lymphocyte counts were studied in 50 patients before they received any therapy for their conditions: 16 had carcinoma of the breast; 8 soft tissue sarcoma; and 26, benign disease. Among 11 males, the six patients with advanced soft tissue sarcoma had significantly lower percentages of T lymphocytes than five patients with benign conditions. And six female patients with advanced breast cancers had significantly higher numbers of B lymphocytes than the 10 patients with early breast malignancies. Pertinent reports in the literature were reviewed and discussed. Presently, the clinical and prognostic values of quantitative study of T and B lymphocytes remain to be defined.


A scanning electron microscopic and immunologic study of B-cell lymphosarcoma cells in cerebrospinal fluid. Homogenous populations of malignant cells from the spinal fluids of two patients with metastatic lymphosarcoma were studied immunologically and by scanning electron microscopy (SEM). The malignant cells in both spinal fluids were shown to be of B-cell origin. The surface morphology of these cells was variable with the majority of the cells having numerous microvilli and a minority having few or no microvilli. This appears to be the first report on the surface ultrastructure of B-lymphosarcoma cells suspended in spinal fluid.


Thymus-dependent and nonthymus-dependent lymphocytes in patients with sarcomas. Several measurements indicate that cellular and humoral immunity is depressed in patients with sarcoma; but whether such observations reflect diminished numbers of lymphocytes and their subpopulations is not known. We determined the numbers of peripheral leukocytes, lymphocytes, rosette-forming thymus-dependent cells (T cells), and nonthymus-dependent SmIg-bearing cells in 134 patients with various mesenchymal sarcoma and in 47 normal controls. In all of these determinations, patients and controls gave similar values. Furthermore, values for patients with or without tumor, with or without metastasis, and with bone or soft-tissue sarcomas were similar. All 134 patients were studied prospectively, and the 84 who remained free of disease after 6 months had virtually identical values to the 50 who showed disease progression.


Hemodynamics at rest and during exercise in 222 patients with coronary heart disease before and after aorto-coronary bypass surgery. In 222 patients with coronary heart disease hemodynamics at rest and during exercise were measured before and after aortocoronary bypass surgery. A total of 552 grafts were constructed, i.e. an average of 2.47 grafts per patient. Only 10.8% of the patients had a 1-vessel-disease, 59.2% had a 3-vessel-disease. 10.8% of the patients were provided with one graft, 49.7% got 3 or 4 grafts. In 92.8% of the patients the r. desc. ant. was significantly stenosed, and in 94.2% this vessel has been provided with a graft. Preoperatively only 7 patients had no angina pectoris during exercise (bicycle ergometer in supine position, each load lasting 6 min), postoperatively 154 patients have been completely free of angina pectoris. The preoperative angina pectoris-free exercise tolerance was 27.4 +/- 27.4 W (means +/- SD), postoperatively it was 76.5 +/- 33.8 W. The largest increase of exercise tolerance was observed in patients with a 3-vessel-disease (208%). Preoperatively only 10.1% had normal values of pulmonary wedge pressure and cardiac output at rest and during exercise, postooperatively 51.5%. The postoperative normalization of hemodynamics depends on the number of vessels involved (1-vessel-disease 86%, 3-vessel-disease 39.8% normalization) and on the status of the left ventricle (without a previous transmural myocardial infarction 68.4%; with a previous myocardial infarction 41.0%). The effects of revascularization on myocardial ischemia can be evaluated by measurements of pulmonary wedge pressure and cardiac output at rest and during exercise.


Proliferation of Buffalo rat liver cells in serum-free medium does not depend upon multiplication-stimulating activity (MSA). A line of Buffalo rat liver cells (BRL 3A) that multiplies in the absence of serum produces a family of polypeptides termed MSA that can partially satisfy the serum requirement for growth of chick embryo fibroblasts. Temin, Pierson and Dulak (1972) proposed that BRL cells multiply in serum-free medium because they produce MSA. This does not appear to be the case. We have studied three BRL cell lines: 3A2 and 3A have diverged from the same original isolate from normal liver; 61t is a spontaneous transformant of a different isolate. All three cell lines showed a 10 fold increase in cell number during 5 days in serum-free medium. However, 3A-conditioned medium stimulated 3H-thymidine incorporation into DNA in chick embryo fibroblasts and human skin fibroblasts; 3A2- and 61t-conditioned media did not. After ion-exchange chromatography or gel filtration of the conditioned media and measurement of MSA by 3H-thymidine incorporation or radioreceptor assay, MSA again was found in the 3A medium but not in the 3A2 or 61t media. The absence of MSA in the 3A2 and 61t media was not due to inactivation of MSA by these two cell lines. Addition of partially purified MSA to 3A2 cells did not increase their multiplication rate in serum-free medium. We conclude that the ability of the BRL cells to multiply in serum-free medium is independent of the level of MSA in the medium.


Enhancement of the therapeutic effectiveness of methotrexate and protection of normal proliferating tissues with purines and pyrimidines. Mice can be protected against the toxicity arising from lethal doses of methotrexate with purine and pyrimidine combinations, but not with pyrimidine alone. Furthermore, methotrexate treatment of tumour-bearing mice, conjunction with purine/pyrimidine protection, can be more effective than conventional metotrexate/folinic acid treatment.


[Digestive hemorrhage as a complication of a pancreatic pseudocyst]. A study was made at the Padua University Second Surgical Clinic--located in Verona--on four cases of digestive haemorrhage in patients with pancreatic pseudo-cysts. The psysiopatological, diagnostic and therapeutic problems inherent in this pathology are discussed. In particular, attention is drawn to the need for and difficulty of marking a correct diagnosis in good time, and the utility, for such purpose, of endoscopic examinations and selective arteriography of the coeliac tripod and the superior mesenteric artery. Lastly, the various possibilities that surgery offers in the treatment of this disease are mentioned.


Vision screening of illiterate populations. To assess the amount of reduced vision in a population is an important public health matter, especially in areas where blinding diseases are endemic. Testing visual acuity is, however, a complex problem when a major part of the population is illiterate. The best-known test of vision is the E-test, but this produces the problem of untestability in illiterate populations.The introduction of the Sjögren hand-test as an alternative to the E-test for vision screening of unselected illiterate populations in West Africa resulted in a highly significant reduction of untestability. For certain vision levels it is possible to correlate the results of the hand-test directly with those of the E-test. The hand-test is less well defined than the E-test, but has important advantages for the purpose of vision screening of illiterate populations.


Isolation and characterization of influenza A viruses from avian species in Hong Kong. Surveillance of apparently healthy ducks, chickens, and geese at a poultry dressing plant in Hong Kong yielded 51 haemagglutinating viruses 25 of which were influenza A viruses. Of these, 24 were subtyped into 13 combinations based on haemagglutinin and neuraminidase surface antigens. Of the 13 different influenza A viruses isolated, 7 possessed combinations of haemagglutinin and neuraminidase subunits that have not been reported previously-i.e., Hav2N1, Hav2Nav5, Hav4N2, Hav7N2, Hav7Nav1, Hav7Nav3, and Hav7Nav6. Four of the isolates were non-avid: they were not neutralized by antisera to any of the reference subtypes of influenza A viruses, yet antisera to each isolate inhibited both that virus and a known reference strain. The large number of combinations of haemagglutinin and neuraminidase and the isolation of two different influenza A viruses from one duck suggests that recombination may be occurring in nature.


Measles immunity in the first year after birth and the optimum age for vaccination in Kenyan children. Collaborative study by the Ministry of Health of Kenya and the World Health Organization. There is still controversy about the optimum age for measles vaccination in developing countries, where the incidence of measles infection is higher in the first few months of life than it is in developed countries. This study was undertaken to collect reliable data in order to determine the optimum age for mass vaccination programmes. Haemagglutination inhibition (HI) antibodies were titrated periodically from birth to one year of age in children who were given the vaccine at different ages, between 5 and 9 months. It was found that 90% of children no longer have their maternal antibodies at 7-8 months of age, precisely at the period that the incidence of measles begins to rise sharply. Almost all children showed HI seroconversion when vaccinated at 71/2 months (or later, but not before), even if a low level of maternal antibody still persisted when the vaccine was given. These data show that there is an advantage in carrying out measles vaccination at 71/2 months of age in countries with conditions similar to that of Kenya. The duration of post-vaccinal immunity beyond one year of age has not been studied, but it can reasonably be expected that immunity after one vaccination can last for at least 3-5 years, thus exceeding the period when African children are most exposed to malnutrition.


The serological response to Yersinia pestis infection. Passive haemagglutination antibody titres to Fraction I antigen of Yersinia pestis were plotted against day of clinical illness in 82 patients in Viet Nam. A rise was evident by day 5 with a peak at day 14, after which a plateau occurred. In contrast to all other patients, 2 patients with recurrent infections had elevated titres at the time of admission which decreased significantly during convalescence.


BCG vaccination by bifurcated needle in a pilot vaccination programme. The bifurcated needle technique for BCG vaccination was compared with intradermal injection in a mass BCG and smallpox vaccination programme in Afghanistan. In all population groups the bifurcated needle technique produced substantial tuberculin sensitivity, which however was significantly inferior to that following intradermal injection. The advantages of the bifurcated needle technique did not result in a significant increase in vaccination coverage and output. Under the present circumstances vaccination by bifurcated needle would probably be more expensive than intradermal vaccination.


Causes of perinatal mortality in an African city. Little has been published about the causes of most of the deaths responsible for the high perinatal mortality rates characteristic of preindustrial, urban societies. The present study searched for answers in one such society by identifying the causes of death in a large-scale analysis of perinatal mortality in Addis Ababa, Ethiopia. A 72% autopsy rate was achieved when 1019 postmortem examinations were performed for 1424 consecutive perinatal deaths. The overall perinatal mortality rate was 65.3 per thousand live births. The ratio of stillbirths to neonatal deaths was 2.7:1, indicating that maternal factors were dominant in causing the deaths. One-third of the deaths were due to amniotic fluid infections, 15% to obstructed labour, 8% to abruptio placentae, and the rest to more than 20 other specific disorders.


Effects of the injectable contraceptive depot medroxyprogesterone acetate in Thai women with liver fluke infestation: results after six months. The effect of the three-monthly injectable contraceptive depot medroxyprogesterone acetate (DMPA) on liver and lipid function was assessed in Thai women with liver fluke (Opisthorchis viverrini) infestation, DMPA administration being started in the immediate postpartum period. Immediate postpartum IUD and sterilization acceptors with fluke infestation were recruited as a comparison (control) group for the fluke-positiv DMPA acceptors. Comparable groups of fluke-negative acceptors were recruited in an area of Thailand free of liver fluke transmission. Results are presented for the first 6 follow-up months for 170 DMPA and 177 control fluke-positive subjects and for 153 DMPA and 150 control fluke-negative subjects. Small and similar increases occurred in each of the four groups for alanine amino transferase, isocitrate dehydrogenase, and total bilirubin levels while aspartate amino transferase levels changed less in the DMPA groups than in their respective control groups. None of the subjects in either DMPA group had clearly abnormal results in these tests at 6 months. Alkaline phosphatase, cholesterol, and triglycerides levels were markedly lower in each group at 6 months than in the puerperal specimens. There was a greater decrease in triglycerides levels in both DMPA groups than in their respective control groups. However, the decrease in the alkaline phosphatase and cholesterol levels was greater only in the fluke-positive DMPA group than in the fluke-positive control group. None of these biochemical results were related to differences in age, parity, or lactation status between the groups. The results indicate that DMPA did not cause any early deleterious effects in the metabolic factors studied in women with liver fluke infestation.


Simplified field assessment of nutritional status in early childhood: practical suggestions for developing countries. This paper proposes a simple and inexpensive method for the field assessment of certain objective indicators of nutritional status in children of preschool age. It emphasizes the need for statistically valid sample selection and presents a design for randomly selecting 30 children from each of 30 village sites in each region for which quantitative inferences are to be made, the main purpose being to estimate the prevalence of protein-energy undernutrition and anaemia. The need to train indigenous paraprofessional workers as assessors and periodically to control their accuracy is stressed. The method used is limited to an estimate of the location and magnitude of common childhood malnutrition and it is recommended that it be supplemented by detailed ecological analysis to determine causal factors and propose remedial action.


Double-antibody radioimmunoassay for factor VIII-related antigen. A plasma protein required for the support of ristocetin-induced platelet aggregation was isolated from antihemophilic factor concentrate and radiolabeled with 125I. A double-antibody radioimmunoassay was developed, with use of specific rabbit anti-VIII related antigen serum and goat anti-rabbit globulin. The assay is sensitive, reproducible, and technically simple to perform. Values obtained in normal subjects ranged from 0.65 to 1.53 units, similar to our normal range for VIII coagulant activity (0.67-1.43 units). However, normal or increased values of VIII-related antigen were observed in VIII coagulant-deficient hemophiliacs. Also, concentrations of VII-related antigen significantly exceeded coagulant concentrations in several patients with liver disease or disseminated intravascular coagulation, or both. Of a broad selection of congenital coagulation disorders examined, only patients with von Willebrand's disease had decreased VIII-related antigen concentrations, and these corresponded to the lowered concentration of ristocetin cofactor in the patients. In three transfused patients, VII-related antigen values correlated with the concentration of the cofactor. Our results suggest that the radioimmunoassay of VIII-related antigen is a simple and valuable adjunct in the study of patients with clotting abnormalities.


Measurement of antitryptic activity of serum with a centrifugal analyzer. The Selected Method [Clin. Chem. 20, 396 (1974)] for the enzymatic assay of alpha1-antitrypsin in serum has been adapted for use with the E.N.I.-GEMSAEC. With alpha-N-benzoyl-D,L-arginine-p-nitroanilide as substrate, the difference between the tryptic activity measured with and without addition of serum in the same run has been used to calculate the trypsin-inhibitory capacity. The rate of increase in absorbance at 400 nm of the p-nitroanilide formed, has been evaluated during a reaction time of 140 s. Results correlated well (r = 0.986) for 54 human sera analyzed as described here and by the Selected Method. The adaptation on GEMSAEC can be used in detecting alpha1-antitrypsin deficiency in the newborn.


Plasma lipid changes in the female in aging and the menopause. Results from three population studies. Postabsorbtive plasma lipid levels have been analyzed in studies of the female population in the Swedish city of Gothenburg. The population samples cover the ages 15, 23, 30, 38, 40, 45, 46, 50, 54, 60 and 70. In some groups data are available from both cross-sectional and prospective longitudinal studies. Cross-sectional studies showed a successive increase in total plasma cholesterol from 15 to 54 years of age. Longitudinal studies showed a further increase at least up to 60 years of age. The triglyceride level remained rather constant between 15 and 30 years of age, then increased but was again rather similar in the 40--50-year olds and reached the highest level in the 60- and 70-year samples. The total phospholipid curve was approximately parallel with the cholesterol curve at younger ages but nearer the cholesterol curve at higher ages. The composition of the plasma phospholipid fraction apparently changed at the ages around the menopause, mainly due to an increase in the proportions of lecithin and lysolecithin. Data available allowed considerations concerning possible influences on plasma lipids by age, by changes in dietary habits and body composition, as well as by gonadal steroid hormones.


Alpha-I-antitrypsin (Pi) phenotypes in Lyon, France: departure from Hardy-Weinberg equilibrium. The validity of the Hardy-Weinberg equilibrium for Pi phenotypes was tested in a population of 1,653 randomly selected, healthy blood donors in Lyon, France. A significant departure from the equilibrium was found, with an excess of FF and SS homozygotes and a deficiency of FM heterozygotes.


Persisting lymphocyte deficiences during remission in Hodgkin's disease. Blood lymphocytes from nineteen patients with Hodgkin's disease were tested in vitro before and after treatment. The patients were retested in complete and unmaintained remission at least 15 months after termination of radiotherapy. All patients except two had been treated with total nodal irradiation. The lymphocyte-DNA synthesis induced by concanavalin A (Con A) and PPD was poor and the spontaneous DNA synthesis was increased in untreated patients. Most patients had a T lymphocytopenia before treatment. After irradiation the total lymphocyte counts were reduced drastically. The number of T lymphocytes was particularly low, though the number of B lymphocytes decreased as well. However, the lymphocyte response to Con A and PPD remained low and unchanged. The results may suggest a persisting immunodeficiency in Hodgkin's disease as reflected by the lymphocyte response to Con A and PPD.


Relationships between antibodies to native DNA and glomerulonephritis in systemic lupus erythematosus. Anti-native DNA antibodies have been evaluated in forty-six lupus patients' sera for antigen-binding capacity, affinity and precipitating activity. Diffuse proliferative glomerulonephritis was significantly correlated with the presence of high serum level of free anti-native DNA antibodies. Weak affinities were more often found in patients with than without glomerular changes but several patients had high-affinity anti-DNA antibodies and severe glomerulonephritis. No correlation was found between anti-DNA antibody-precipitating activity and renal lesions.


Detection and characterization of DNA-anti-DNA complexes in a patient with systemic lupus erythematosus. DNase digestion of SLE serum, with consequent release of bound DNA antibody has been proposed as a method for the direct demonstration of circulating DNA-anti-DNA complexes. In the present studies on the serum of a girl with active SLE nephritis, circulating DNA-anti-DNA complexes were demonstrated at the precise time of relapse of SLE nephritis. Ultracentrifugation showed that these complexes were of low molecular weight.


Spontaneous lymphocyte transformation in chronic lymphocytic leukaemia. In ten patients with CLL and in eight controls T lymphocytes were separated out by the rosetting method. The in vitro reactivity of these cells and of unseparated lymphocytes was studied in cultures without stimulants. A net increase of spontaneous lymphocyte transformation was found in the populations enriched with T cells in patients with CLL with respect to analogous populations in the controls. This increase is interpreted as a cell-mediated immunological reaction and due to T-cell sensitization towards the neoplastic B cells.


Suppressor T cells prevent experimental autoimmune encephalomyelitis in mice. Immune suppression (immunoprotection) in experimental autoimmune encephalomyelitis (EAE) was studied in (SJL X BALB/c)F1 mice using inocular of mouse spinal cord homogenate (MSCH), or mouse basic protein of myelin (M-BPM), in Freund's incomplete adjuvant (FIA). Such immunization specifically recruited lymphoid cells which markedly suppressed the capacity of effector lymph node cells from appropriately immunized syngeneic mice to transfer adoptively EAE. Suppression was demonstrable with transfer of bone marrow and spleen cells, but not with lymph nodes or thymus cells. Adoptively transferred suppression was maximal when cells were injected 9-30 days after the suppressive injection. Inhibition of EAE by suppressor cells was specific for the relavant antigen BPM, and required viable cells. Treatment of cells with anti-Thy-1 serum before transfer abolished their suppressor activity. After adoptive transfer of suppressor cells into syngeneic recipients subsequently immunized for EAE, there was inhibition of EAE and reduced cell-mediated immune response to BPM as judged by macrophage migration inhibition assays. Hence, in mice at least, immuno-protection against EAE is explicable by recruitment of suppressor T lymphocytes with the dual capacities of inhibiting development of effector T cells after antigenic stimulation, and of blocking their damaging effects on the antigen in the central nervous system.


Suppressor cells and loss of B-cell potential in mice infected with Trypanosoma brucei. The functional changes in splenic lymphoid populations from mice infected with T. brucei strain S42 were studied throughout the 3 weeks of infection. Within a week of infection, proliferation of B and T cells profoundly increased as shown by 3H-labelled thymidine incorporation and fluorescent staining of surface Ig; the spleen cells secreted high levels of both IgM and IgG immediately cells were put into culture; but with progressing infection this Ig production declined. The early effect on T cells was reflected by lack of responsiveness to PHA. B-cell potential was studied in low-density cultures treated with lipopolysaccharide (E. coli). Normal spleen cells proliferate extensively in these cultures with subsequent secretion of IgG as well as IgM. The ability to proliferate and produce Ig in response to LPS was severely depressed by day 7 and almost totally absent by day 12 of infection. Removal of T cells from the spleen cells obtained early in infection partly restored the response to LPS but as the infection neared its fatal end, B-cell potential appeared to become exhausted. Macrophages obtained from infected mice even early in infection profoundly depressed the ability of normal spleen cells to proliferate and secrete immunoglobulin in LPS cultures. The general immunodepressing effect of trypanosomes can be attributed to clonal exhaustion of B-cell potential caused by an undefined blastogenic stimulus from the parasites which may operate at least in part by the generation of suppressive T cells and macrophages.


The mechanism of action of cyclophosphamide on the nephritis of (NZB x NZW)F1 hybrid mice. The effects of cyclophosphamide an antinuclear antibody levels, immune complex deposition in renal glomeruli, glomerular cell proliferation and glomerulosclerosis in the (NZB x NZW)F1 hybrid mouse have been investigated in order to better understand the mechanism of action of this drug on nephritis. Five groups of mice were injected daily with this agent (15 mg/kg) by the i.p. route over a 2-month period. Treatment periods began at 1, 3, 5, 7 and 9 months. Each group was killed at the end of the treatment period. During the last week [3H]Tdr(1 mu Ci/g) was injected daily i.p. Immune complex deposition was measured by quantification of glomerular immunofluorescent staining in both capillary loops and mesangium. Glomerular cell proliferation was quantified by measurement of the number of glomerular cells which incorporated [3H]Tdr during in vivo labelling. The number of [3H]Tdr-labelled cells in each of 100 glomeruli was counted using kidney autoradiographs of whole kidney slices. This technique provided a very reproducible and quantitative index of glomerular cell proliferation. Glomerulosclerosis was measured by determining percentage of PAS-positive material in each of 100 counted glomeruli. Immune complex deposition in the kidney preceded the glomerular cell proliferation. The proliferation reached a peak at 9 months of age. Glomerulosclerosis gradually increased with age until the animal's death. Cyclophosphamide significantly decreased antinuclear antibody levels, glomerular cell proliferation, and immunoglobulin staining in the glomeruli. Progression of glomerulosclerosis was significantly arrested. The present findings demonstrate a very significant effect of cyclophosphamide on three important pathological changes which occur in the nephritis of the (NZB x NZW)F1 hybrid mouse and provide information about possible mechanisms underlying these changes.


Selective expression of separate I-region loci in functionally different lymphocyte subpopulations. Determinants controlled by separate loci mapping in the I region of the H-2 gene complex are selectively expressed on subpopulations of lymphocytes which play different roles in generating humoral responses. The Ia-4 locus, which marks a new I subregion, designated I-J, controls determinants found on allotype suppressor and normal (nonsuppressed) T lymphocytes. These determinants are not present on helper T lymphocytes or B lymphocytes. In contrast, the Ia-1 locus, which marks the I-A subregion, controls determinants present on B lymphocytes but not on suppressor or helper T lymphocytes. Another locus (subregion not known) controls determinants which distinguish helper T lymphocytes from suppressor T lymphocytes. Such selective expression suggests that the products of these loci may plan an integral role in lymphocyte interactions.


Recognition of alterations induced by early vaccinia surface antigens and dependence of virus-specific lysis on H-2 antigen concentration on target cells. Specific recognition of antigens by cytolytic T lymphocytes sensitized to vaccinia virus was tested by monolayer adsorption. Adsorption was possible only on monolayers also expressing syngenic H-2 as viral antigens present during the sensitization phase. Vaccinia-virus-infected target cells were subjected to papain and neuraminidase treatment. H-2 antigenic determinants could be removed by papain treatment. Due to virus-specific inhibition of host-cell protein synthesis, reexpression of H-2 antigenic determinants did not take place, but viral surface antigens were resynthesized. Susceptibility of target cells to T-cell-mediated lysis was decreased after papain treatment. Substrains of vaccinia virus were used in order to define the minimal changes induced by vaccinia virus necessary for T-cell sensitization in vivo and target-cell lysis in vitro. When the immune response to a conditioanl lethal mutant strain of vaccinia virus was investigated, it could be demonstrated that expression of early surface antigens is sufficient for induction of the cellular immune reactions. These data were confirmed by inhibition studies with virus-specific antisera.


Antigen activation of T lymphocytes: influence of major histocompatibility complex. There is considerable evidence that T-cell activation to soluble antigens occurs only if this is processed by macrophages and displayed appropiately on the cell membrane in association with products of the genes of the MHC. The genes responsible differ according to the cells and antigens involved. For cytotoxicity, targets and killer T cells must share K- or D-region gene products. For delayed-type hypersensitivity to FGG in mice, I-A identity is necessary; for DNFB, identity at either the I, K, or D region is sufficient. Experiments using three different approaches do not support the notion that these genetic constraints are due to the necessity for the T cell and stimulator cell to match an identical gene product or cell-interaction molecule. Rather, they favor the hypothesis that there are receptors on the activated T cell which recognize antigen and products of genes of the MHC. The implications of the results are discussed in terms of (1) different T-cell subsets, (2) the mode of action of Ir genes, and (3) the possible parallel evolution of T-cell receptors for antigen and gene products of the MHC.


An information system for clinical recording, administrative decision making, evaluation, and research. An information system suitable for clinical, administrative, and research uses is described. The source document is a modified problem-oriented record in which progress on each problem can be measured by a goal attainment scaling technique during therapy, at termination, and at follow-up. The model deals with the problem of how specific the goals should be by establishing two evaluation procedures. One procedure is based on predefined long-term general goals of therapy. The other procedure deals with therapist defined short-term goals and procedures.


Oral contraceptives and hypertension.

Oral contraceptives and hypertension. A variety of studies have noted that the use of oral contraceptives generally leads to mild increases in blood pressure which are usually reversible when the medication is discontinued. Representative data from the Walnut Creek Contraceptive Drug Study and the Royal College of General Practitioners Study concerning the magnitude of excess risk and relation to duration of use and pull content are shown. Preliminary data from women, aged 25-34 years, taking multiphasic health checkups in Oakland and San Francisco, suggest that black as well as white women are susceptible to this side effect of oral contraceptives. A method is given for estimating the proportion of hypertensives among a population of young women that is attributable to oral contraceptive use. Although the risk of pull-induced hypertension is small for the average user, oral contraceptives appear to be an important identifiable cause of hypertension in samples of women studied.


Gastrointestinal hemorrhage in patients in a respiratory intensive care unit. Significant gastrointestinal hemorrhage occurred in 20% (20) of the patients in a respiratory intensive care unit. Risk factors significantly associated with the development of gastrointestinal hemorrhage included (1) the principal respiratory diagnosis of adult respiratory distress syndrome; (2) increasing numbers of days on a ventilator, days in the respiratory intensive care unit, and total days of hospitalization; and (3) the development of thrombocytopenia. Factors not associated with an increased risk of gastrointestinal hemorrhage were the age and sex of the patient, the respiratory diagnosis of chronic obstructive pulmonary disease, and the use of therapy with either heparin or corticosteroids. Routine prophylactic administration of antacids was associated with a decreased incidence of hemorrhage. The mortality of bleeders was significantly greater than that of nonbleeders.


Mercuric chloride-induced anti-glomerular basement membrane antibodies in the rat: genetic control. Mercuric chloride induces anti-glomerular basement membrane antibodies in the Brown-Norway rat. Various other inbred rat strains (Lewis, Wistar AG, August, PVG/c) were not found to be able to produce such antibodies under the same experimental conditions. Hybrids (F1, F2 and F1 x LEW) were bred from Brown-Norway and Lewis rats and injected with mercuric chloride. It has been demonstrated that the induction of anti-glomerular basement membrane antibodies by mercuric chloride in these crosses is under genetic control. The response was found to depend on two or three genes one of which was H-1-linked. The negative results obtained with L.BN congenic rats were in complete agreement with this conclusion.


Increase in T cell mitogen responsiveness in rat thymocytes by thymic epithelial culture supernatant. Supernatants from rat thymic epithelial cultures (TES) increased [14C]thymidine incorporation into rat thymocytes stimulated with phytohemagglutinin (PHA) or concanavalin A, whereas supernatants from nonthymic control tissue cultures had no effect. It was demonstrated that the increased mitogen responsiveness was at least partially due to an increase in the number of responsive cells. Furthermore, our data suggest that the target cell for TES resides among the cortisone-sensitive population of thymocytes. The continuous presence of TES provided the best conditions for enhancement of mitogen stimulation. The addition of TES had very little effect on mitogen-stimulated spleen and lymph node lymphocyte cultures. The data suggest that, in the presence of TES, a cortisone-sensitive, PHA-nonresponsive subpopulation of rat thymocytes is induced to respond to PHA.


Effect of tumor cells on the generation of cytotoxic T lymphocytes in vitro. I. Accessory cell functions of mouse tumor cells in the generation of cytotoxic T lymphocytes in vitro: replacement of adherent phagocytic cells by tumor cells or 2-mercaptoethanol. In agreement with previous reports, the primary in vitro response to alloantigens has been shown to be dependent on the presence of macrophages (Mphs). Splenocytes extensively depleted of adherent phagocytic cells did not generate cytotoxic T lymphocytes, and this activity could be completely restored by small numbers of adherent peritoneal cells (accessory cells). Either P388D1 (Mph-like tumor), P388 ("null" tumor) or P815 (mastocytoma) tumor cells, or 2-mercaptoethanol, could completely replace the accessory function normally mediated by accessory cells. These tumor cells did not non-specifically "enhance" the cytotoxic activity generated with normal nondepleted spleen cells. The restored cultures maintained killing specificity to H-2 targets which was mediated by effector T cells as shown by sensitivity to anti-theta and complement. Therefore, Mphs seem not to be the sole cells capable of mediating an accessory function in a primary response to alloantigens in vitro.


Cellular events in protein-tolerant inbred rats. IV. The mechanism of immunogen-maintained tolerance. The ability of immunogens to maintain or extend a state of unresponsiveness was investigated in unbred rats using a human serum albumin (HSA) model of tolerance. Rats initially challenged with immunogen within two weeks of high or low dose tolerance induction by tolerogen (soluble HSA) remained hyporesponsive even a year and a half later and in some cases became less responsive following a subsequent challenge. An inhibitory effect of immunogen on escape from tolerance was formally demonstrated: in comparison with an unchallenged group, tolerant rats which received a second immunogen challenge 6 months after the first, synthesized less antibody; this antibody underwent a gradual decline in affinity after each challenge which suggested that higher avidity B cells were progressively lost. In addition, immunogen-maintained tolerant rats (a) had demonstrable helper T cell activity among their thoracic duct lymphocytes on adoptive transfer, (b) did not produce a significant increase in antibody synthesis after receiving peripheral T cells and (c) provided no evidence that suppressor cells were playing a role. The results suggested that the mechanisms of tolerance induction by tolerogen and tolerance maintenance by immunogen are fundamentally different.


H-2 gene complementation in cytotoxic T cell responses of female against male cells. H-2 gene-dependent nonresponsiveness and responsiveness to H-2-matched male cells can be observed in cytotoxic assays in several mouse strains. H-2 genes of several low responder strains can complement each other in cis or trans position to produce high responders. One of the H-2 gene products functions at the level of antigen-presenting cells (stimulators, targets), the other must be expressed in the responding T cell population.


Accessory cell dependence of lectin-induced proliferation of mouse T lymphocytes. Mouse lymph node cells were exposed to carbonyl iron and a magnet to remove phagocytic cells, and passed over Sephadex G-10 and nylon wool and incubated for 12 h on plastic to remove adherent cells and their precursors. More than 99% of the cells in this macrophage-depleted population (which constituted 3-5% of the starting population) were Thy-1+ and Ly-1+, while less than 2% were Ly-2+. These cells usually did not synthesize detectable amounts of DNA when cultured with concanavalin A and responded poorly to phytohemagglutinin. These proliferative responses were completely reconstituted by small numbers of syngeneic or allogeneic peritoneal cells, purified peritoneal macrophages or cells from tertiary cultures of mouse embryo 'fibroblasts', but not by 3T3 cells, P815 mastocytoma cells or Nulli SCC-1 embryonal carcinoma cells, or by 2-mercaptoethanol. The reconstituting peritoneal cells were Thy-1--, Ia+ and present in nu/nu mice; although they had to be alive to reconstitute, they did not have to divide. These results are consistent with the hypothesis that T cell proliferation induced by lectins, like that induced by antigens, may involve the dual recognition of stimulating ligand in association with major histocompatibility complex (Ia) determinants.


Differential antagonism by chlorpromazine and diazepam of frog motoneurone depolarization induced by glutamate-related amino acids. The effects of some central depressant drugs on amino acid-induced depolarization of motoneurons have been determined in the isolate hemisected frog spinal cord. Motoneuron depolarization was recorded from ventral roots and measurements were made in the presence or absence of procaine or tetrodotoxin to minimize indirect effects of both drugs and amino acids. Chlorpromazine (0.05-0.1 mM) and diazepam (0.5 mM) produced a similar differential pattern of depression of amino acid-induced depolarizations. Responses induced by L-homocysteate were markedly antagonized by these drugs, while responses to quisqualate were unaffected. L-Aspartate-induced responses were antagonized more than L-glutamate-induced responses. This pattern of antagonism resembles that previously described for Mg2+. In contrast, pentobarbital (0.1 or 0.3 mM), and the inhibitory amino acids GABA and beta-alanine (0.5-1.0 mM), depressed amino acid-induced responses in a more uniform manner. The differential effects observed with chlorpromazine and diazepam provide further support for the possibility that responses to excitant amino acids structurally related to L-glutamate may have different underlying mechanisms.


The nucleocytoplasmic distribution of 3-O-methylglucose in the amphibian oocyte. Ultra-low temperature microdissection was employed to determine the in situ nucleocytoplasmic distribution 3-O-methylglucose in the amphibian oocyte. The nucleocytoplasmic partition ratio, Kn/c, for this non-metabolizable monosaccharide was 1.54 +/- 0.08. The observed asymmetry could be explained by the differential solubility of the solute in the water of the nucleoplasm and the cytoplasm.


Comparative studies on the trichomonacidal activity of 5-nitroimidazole-derivatives in mice infected s.c. or intravaginally with T. vaginalis. The trichomonacidal efficacy of metronidazole, tinidazole, nimorazole and ornidazole was studied in mice infected s.c., or for comparison purposes intravaginally. In both test systems, the drugs revealed nearly the same order of efficacy, whereby the compounds showed a marked decrease of activity when analyzed in s.c. infected mice.


An initial phasic depolarization exists in the receptor potential of taste cells. When frog taste cells were stimulated by varying salts after adaptation to water, quinine or acetic acid, a phasic depolarization appeared initially in the receptor potential of taste cells. The initial transient depolarization may be related to the enhancement of an initial phasic response in the taste nerve.


Carbon dioxide sensitivity of pulmonary receptors in the frog. Pulmonary mechano-receptors have been found in the frog lung that are sensitive to CO2 concentrations in the lungs and airways comparable to the physiological levels recorded in frogs. These results support the suggestion that a pulmonary receptor with distinct mechano- and chemosensitive properties may represent the functional precursor of the more specialized pulmonary receptor types which have evolved in birds and mammals.


[Role of creatine phosphokinase systems in regulating the force of myocardial contraction in frog ventricles]. Creatine phosphate (10 and 20 mM) increased both the force of contraction and action potential duration in isolated frog heart strips. In concentration of 75 mM creatine phosphate increased the force of contraction during first minutes with its following decrease. Action potential duration was also increased during this period. These data suggest existence of receptory sites for creatine phosphate in the heart cells: the myofibrillar creatine phosphokinase system where creatine phosphate increases the force of contraction; and the cellular membranes where creatine phosphate decreases the rate of membrane repolarization. Dinitrofluorobenzene (1.5 mM) completely inhibited contraction of the heart strips and significantly decreased the duration and intensity of action potential. The data obtained are discussed from the view--point of existence of a creatine phosphate pathway for intracellular energy transport in the heart cells.


[Intracardiac nervous regulation of the systolic and diastolic activity of isolated heart ventricle]. In isolated hearts of cats and frogs, sistolic and diastolic pressure in the ventricle, volume speed, speed of contraction and relaxation, and indexes of contractability and elasticity, were estimated during stretching of the venous receptacle. The data showed that the stretching increased both the sistolic and diastolic activities of the ventricle. Shunting off of the intracardial nervous apparatus prevented these interrelationships. The increase in the ventricle sistolic and diastolic activities due to stimulation of mechanoreceptors of the heart venous receptacles can be regarded as the display of a regulatory activity of intracardial nervous system related to changes of the blood volume coming to the venous receptacles.


Endoscopic laser treatment. II. Comparison of the efficacy of high and low power photocoagulation in control of severely bleeding experimental ulcers in dogs. The coagulative efficacy of a "high power" argon laser which delivers 6.5 +/- 1.0 w was compared to that of a "low power" argon laser which delivers 1.0 +/- 0.1 w. The wave-guide angle of divergence was 8 degrees. For the high power laser, the distance between the wave-guide tip and the mucosa varied from 1.0 to 1.5 cm with a delivered power density range of 160 to 487 w per cm2. For the low power laser, the distance between the tip and mucosa varied from 0.7 to 1.3 cm with a power density of 35 to 146 w per cm2. Group A consisted of acute experiments in 6 heparinized dogs in which 51 standard-sized acute gastric ulcers extending into the submucosa were made at laparotomy via a large gastrotomy. Ulcer bleeding rates were quantified into three categories of severity. Within each category, ulcers were randomized to high power, low power, or untreated control. All 19 low power-treated ulcers and 16 untreated controls continued bleeding; 13 of 16 high power-treated ulcers stopped bleeding completely. Representative ulcers were examined histologically. Group B consisted of chronic experiments on 31 ulcers in 7 unheparinized dogs. These ulcers were either treated with high power or left as untreated controls. Dogs were killed and the ulcers were examined histologically at 7, 14, and 28 days. Only rarely did the laser injury penetrate into the muscularis externa. There were no perforations. By 14 days, all lesions were covered with normal surface epithelium. These data encourage further development of congruent to 7.0 w argon laser photocoagulation for eventual clinical use in man.


Morphologic changes in adverse drug reactions in infants and children. The morphologic findings in 102 validated cases of adverse drug reactions classified in the "causative" and "probable" category are reported. All of them are well documented cases of drug associated reactions. The patterns of tissue reactions in these cases are divided into the following categories: inflammation, degeneration and infiltration, hypoplasia and aplasia, hyperplasia, necrosis, circulatory disturbances and vascular abnormalities, and teratogenesis. Degeneration and infiltration constitute the most common tissue reaction pattern present in these cases. The liver is the organ most frequently affected. Anti-infective drugs were associated with the highest number of adverse drug reactions, followed by antineoplastic drugs.


Effect of streptolysin S on human and mouse T and B lymphocytes. The effect of streptolysin S on T and B human and mouse lymphocytes was studied. The dose-dependent inhibition of T cell rosette formation was observed, and this was accompanied by a dose-dependent cytotoxicity. The existence of a difference in the degree of susceptibility of T and B cells to the cytotoxic effect of streptolysin S is shown.


Preparation of arthritogenic hydrosoluble peptidoglycans from both arthritogenic and non-arthritogenic bacterial cell walls. Cell wall lytic enzyme (Kyowa lytic no. 2 enzyme) liberated arthritogenic hydrosoluble peptidoglycans from both arthritogenic and non-arthritogenic bacterial cell walls. From these cell walls, mutanolysin (peptidoglycan-degrading enzyme) also liberated hydrosoluble peptidoglycans which, however, lacked arthritogenicity. Based on the chemical composition of these peptidoglycans, it was suggested that their arthritis-inducing ability depends on a relatively long chain of glycan units that consists of repeated units of N-acetylglucosaminyl-N-acetylmuramic acid. However, the glycan chain lengths on these peptidoglycans appeared to be related to their adjuvancy rather than to an antigen(s) responsible for development of arthritis in rats.


Effect of food on absorption of indoprofen administered orally to man in two dosage forms. The influence of food on the bioavailability of two oral dosage forms (100-mg capsules and 200-mg tablets) of indoprofen, a new propionic acid derivative with marked anti-inflammatory and analgesic properties, has been investigated. Plasma levels and urinary excretion of indoprofen were determined both in the fasting state and after a standard meal in healthy volunteers after administration of two 100-mg capsules (4 subjects) and of one 200-mg tablet (6 subjects). Indoprofen in biological fluids was determined by gas-liquid chromatography. The extent of absorption from tablets was not affected by food as indicated from the values of the total area under plasma level curves and urinary excretion of the drug. The rate of absorption was faster after meal than in the fasting state. The opposite was found for capsules, which showed a slightly delayed absorption after food. The results suggest that food may differently influence the absorption pattern of different pharmaceutical forms of the same drug.


Isolation and characterization of highly purified alpha-1-antitrypsin. Highly purified human alpha-1-antitrypsin (phenotype MM) was obtained by an original method of preparative electrophoresis. The criteria of homogeneity were assured by one arc in crossed immunoelectrophoresis and one band on polyacrylamide gel. A unique N-terminal amino acid (pyroglutamic acid) and a unique C-terminal residue (lysine) were identified. Determined by gel electrophoresis, its molecular weight was 47,000 daltons.


Arginine modification in Kunitz bovine trypsin inhibitor through 1, 2-cyclohexanedione. Arginine residues (5.5 out of 6) of the trypsin-kallikrein inhibitor from bovine organs (Kunitz inhibitor) were selectively modified by reaction with 1, 2-cyclohexanedione in sodium borate buffer, pH 9.0. The modified inhibitor is still highly active in inhibiting trypsin and chymotrypsin at 1:1 inhibitor: enzyme molar ratio and full inhibition was achieved at slightly higher molar ratio. The extent of correct refolding, upon reoxidation, of the reduced, arginine-modified inhibitor is diminished and regeneration of two arginines occurred under the reduction conditions. The stability constants and the standard-free energies of binding of the complexes between trypsin, or chymotrypsin, and the native, the arginine-modified and the reduced and reoxidized arginine-modified inhibitor have been determined from inhibitory assays.


Effect of salt solutions on radiosensitivity of mammalian cells. III. Treatment with hypertonic solutions. V79 Chinese hamster cells were treated with hypertonic solutions of NaCl or KCl and irradiated rat various times before, during, or after exposure to the solution. In solutions of molarities between 0-2 and 0-5 M, the cellular radiosensitivity increases with the molarity of the bathing solution. At these molarities, the hypertonic solution need not be present during irradiation to sensitize cells. Furthermore, radiosensitivity of cells could be increased by exposing cells for longer times to the hypertonic solution before irradiation. At higher salt concentrations (at 1-5 to 1-8 M), significant radioprotection is observed. Survival curve data showed that this protection was characterized by an increase in DO and a decrease in n, while the survival curves of cells sensitized with 0-465 M NaCl or with lower concentrations exhibited mainly changes in DO. The 1-55 M NaCl solution must be present during radiation to give a protective effect. Prolonged exposure to the salt before irradiation reduced the amount of radioprotection afforded by the salt. The results are discussed in terms of the effects of ions on histones, cellular water structure and the cell-aging cycle.


Sensitivity of the vibrios to ultraviolet-radiation. The ultraviolet-inactivation kinetics of a number of strains of Vibrio cholerae (classical), Vibrio cholerae (el tor), NAG vibrios and Vibrio parahaemolyticus were investigated. Statistical analyses revealed significant differences between any two of the four types of vibrio in respect to their sensitivity to U.V.


Split-dose recovery for radiation-induced tumours in rat skin. Tumour-related recovery in rat skin was estimated from the dependence of tumour yield on time between split doses of electron radiation. Tumour yield versus dose was established at nine dose points, and at three points the dose was split into two equal fractions spaced 0-25, 3-2 or 6-3 hours apart. After irradiation the rats were observed periodically for at least 64 weeks, and at death the tumours were examined histologically. The dependence of yield on dose for single doses was consistent with a quadratic function up to a peak yield at about 1600 rad. The effect of split doses on tumour yield depended on the position on the dose--response curve. At the lowest split dose, the yield declined with a half-time of about 1-8 hours. At the intermediate split dose, an initial increase was followed by a decline with a half-time of about 3-9 hours. At the highest split dose, the tumour yield increased with time between exposures. Fractionation-induced increases in tumour yield were explained as a sparing effect on cell lethality, whereas tumour-related recovery per se was indicated at the lower two doses.


The response of mouse skin to combined hyperthermia and X-rays. The effects of combined hyperthermia and X-irradiation were studied in the skin of the mouse ear. Ears were heated for 1 hour by immersion in a waterbath at temperatures ranging from 37 degrees C--43 degrees C. These heat treatments had little visible effect alone, but when combined with X-rays, enhanced the radiation response. Enhancement depended on the degree of heating. When heat was given immediately after X-rays, the radiation dose to cause a given skin reaction had to be reduced by about 10 per cent for 37 degrees C and about 40 per cent of 43 degrees C. The timing and sequence of the two treatments were important. Heat after X-rays was less effective than heat before X-rays. When heat followed X-rays, the enhancing effect was lost completely if the interval exceeded 4 hours. When heat preceded X-rays, the effect was lost more slowly, depending on temperature. The implications of this for the treatment of cancer by combined therapy are discussed.


A comparison of psycholphysical and VECP increment threshold functions of a rod monochromat. Increment thresholds of a rod monochromat were obtained over a wide range of background luminances. Within each experimental session, visual-evoked cortical potentials (VECP's) were computed at several luminance levels above the increment threshold for each background level. The results indicate that as the background level approached 1,000 scotopic trolands (td.), the VECP began to decrease in amplitude and it was unrecordable above 2,000 scotopic td. The VECP increment threshold function was determined with latency used as the criterion response. Good agreement was found between the psychophysical and VECP functions, although the VECP function did show a tendency to underestimate the threshold at the two dimmest background levels. It is concluded that the dramatic drop in the VECP amplitude at high background luminances is a reflection of the saturation of the rod system and that the VECP appears to be a sensitive correlate of the psychophysical data.


[Nystagmus as an objective assessment of the cervical spine syndrome and its treatment (author's transl)]. Nystagmus together with the history, X-ray and palpation findings were used to confirm the diagnosis of the cervical spine syndrome. In patients with this disorder the nystagmus was controlled by manual treatment in 63%. Nystagmus therefore can be used objectively to assess the success of therapy in cervical spine syndrome.


Purification of HLA-linked B lymphocyte alloantigens in immunologically active form by preparative sodium dodecyl sulfate-gel electrophoresis and studies on their subunit association. The HLA-linked B cell alloantigen (p29,34) is composed of two subunits of 29,000 (p29) and 34,000 (p34) molecular weight. The partially purified HLA-linked B cell alloantigen was purified by a final step of preparative sodium dodecyl sulfate-gel electrophoresis. An antiserum was prepared against p29,34 which specifically lysed B lymphocytes. In sodium dodecyl sulfate at 21 degrees, p29 and p34 remained noncovalently associated and retained immunologic activity; subunit dissociation at higher temperatures correlated with loss of immunologic activity. Although the pI values of p29 and p34 are 6.1 and 5.2, respectively, the subunits co-electrofocus under nondenaturing conditions. In addition, cross-linking studies showed the B cell antigen has a (p29)1(p34)1 subunit structure.


The ventriculo-cephalic ratio on computed tomography. A simple method to describe the size of the cerebral ventricules on CT scans is presented.


Portal hypertension and bleeding esophageal and gastric varices: a surgical approach to treatment. Ascites, hepatic encephalopathy, and bleeding esophageal and gastric varices are frequent consequences of hepatic disease and portal hypertension. Variceal bleeding, however, is the most dangerous sequel and the most yielding to surgical intervention. Portal decompression by the shunting procedure is instituted in order to decrease the risk of future gastrointestinal hemorrhage. The presence of hepatic disease and alterations in hepatic blood flow following the procedure present implications for postoperative management of fluid and electrolyte balance, mental status, ventilation, and coagulation.


Pneumonia due to Haemophilus influenzae (H. aegyptius) biotype 3. Haemophilus influenzae (H. aegyptius) biotype 3 was isolated from eye, nasopharyngeal, and sputum cultures of a 23-month-old male and from sputum and transtracheal aspirate cultures of his 39-year-old mother, both with diffuse bronchopneumonia.


Primary bioassay of human myeloma stem cells. The ability to clone primary tumors in soft agar has proven useful in the study of the kinetics and biological properties of tumor stem cells. We report the development of an in vitro assay which permits formation of colonies of human monoclonal plasma cells in soft agar. Colony growth has been observed from bone marrow aspirates from 75% of the 70 patients with multiple myeloma or related monoclonal disorders studied. Growth was induced with either 0.02 ml of human type O erythrocytes or 0.25 ml of medium conditioned by the adherent spleen cells of mineral oil-primed BALB/c mice. 5-500 colonies appeared after 2-3 wk in culture yielding a plating efficiency of 0.001-0.1%. The number of myeloma colonies was proportional to the number of cells plated between concentrations of 10(5)-10(6) and back-extrapolated through zero, suggesting that colonies were clones derived from single myeloma stem cells. Morphological, histochemical, and functional criteria showed the colonies to consist of immature plasmablasts and mature plasma cells. 60-80% of cells picked from colonies contained intracytoplasmic monoclonal immunoglobulin. Colony growth was most easily achieved from the bone marrow cells of untreated patients or those in relapse. Only 50% of bone marrow samples from patients in remission were successfully cultured. Tritiated thymidine suicide studies provided evidence that for most myeloma patients, a very high proportion of myeloma colony-forming cells was actively in transit through the cell cycle. Velocity sedimentation at 1 g showed myeloma stem cells sedimented in a broad band with a peak at 13 mm/h. Antibody to granulocyte colony-stimulating factor did not reduce the number or size of the colonies. Increased numbers of myeloma colonies were seen when the marrow was depleted of colony-stimulating factor elaborating adherent cells before plating. This bioassay should prove useful in studying the in vitro biological behavior of certain bone marrow-derived (B)-cell neoplasia. In addition, systematic and predictive studies of anticancer drug effects on myeloma stem cells should now be feasible.


Measurement of cytoplasmic fluorescence depolarization of single cells in a flow system. We have built a flow system in which we can analyze and sort individual viable cells on the basis of their cytoplasmic microviscosity. The average cytoplasmic microviscosity (or cytoplasmic structuredness) of a cell can be quantitated by exciting fluorescein molecules in the cytoplasm of the cell with a polarized light source and measuring the extent of depolarization of the fluorescent signal. Changes in the state of a cell (e.g., the reception of a signal inducing the cell to differentiate) often appear to be associated with changes in cytoplasmic microviscosity, these changes being detectable within hours of the inducing signal. An example of one such change is presented.


Differentiation of lymphoid cells: the preferential binding of the lipid A moiety of lipopolysaccharide to B lymphocyte populations. Lipid A, prepared from lipopolysaccharide, was labeled with 125 I. Such iodinated lipid A possesses the full mitogenic activity of untreated lipid A. Comparison of the 125 I-lipid A-binding activity of splenocytes and thymocytes from the same rabbit revealed that the extent of labeling of splenocytes was 10 to 20 times greater than that observed with an equivalent number of thymocytes. A similar preferential binding was detected in comparing cells in mouse and rat. Spleen populations depleted of adherent cells were essentially unaltered with regard to binding when compared to the original population. In addition, spleen cell populations enriched for thymus-derived cells (T cells) exhibited a marked loss of specific binding activity. On the other hand, spleen cell populations enriched for bone marrow-derived cells (B cells) exhibited the expected binding. The difference in binding behavior of B and T cell-enriched populations was confirmed by using three independent techniques to separate B and T cells. These findings are consistent with the mitogenic specificity of lipid A toward B cells rather than T cells and suggest that the observed cellular specificity resides in an early event in mitogenesis, i.e., binding of the mitogen.


Suppressor cells in mice infected with Trypanosoma brucei. Within 2 to 3 days of infection with Trypanosoma brucei strain S42, the ability of spleen cells from infected CBA mice to mount a primary in vitro antibody response to sheep red blood cells (SRBC) is profoundly reduced, and suppressor cells are generated as detected by cell mixture experiments. Suppressor cell activity lies in the T and adherent cell compartments of spleens from infected mice, but not in the B cell compartment, although antibody responses to a thymus-independent antigen, DNP-Ficoll, are significantly reduced. Suppression of antibody responses of normal spleen cells depends on viable cells from infected mice. The trypanosome, itself, plays no direct role in suppression, and we have ruled out the possibility of antigenic competition as a mechanism of suppression. Our data is consistent with the model of suppressor T cells induced by concanavalin A mitogenesis. We hypothesize that trypanosome antigens may directly stimulate T cells with the concomitant release of factors with affinity for macrophage surfaces thus becoming suppressive for T and B cell responses.


Experimental allergic encephalomyelitis: enhancement of cell-mediated transfer by concanavalin A. Adoptive transfer of experimental allergic encephalomyelitis (EAE) with splenic lymphocytes from Lewis rats sensitized to myelin basic protein (BP) was potentiated by incubation of the cells in vitro with concanavalin A (Con A). Spleen cells of donors which had recovered from EAE also transferred the disease readily after activation by this procedure. In contrast, the transfer of activity of lymph node cells was not altered. We conclude that during the course of EAE a population of T cells with immunologic memory for BP is generated and persists in the spleen. Incubation with Con A activates these cells and results in marked enhancement of their ability to transfer the disease.


Human suppressor T cells induced by concanavalin A: suppressor T cells belong to distinctive T cell subclasses. Human peripheral blood lymphocytes were stimulated by concanavalin A (Con A) and then evaluated by their suppressive activity for thymus-derived (T) cell- and bone marrow-derived (B) cell-proliferative responses to mitogen and allogeneic cells. Con A-activated T cells markedly suppressed these responses, but Con A-activated B cells failed to demonstrate suppressor activity. Discontinuous bovine serum albumin (BSA) density gradient separation of T cells which had been activated by Con A demonstrated that a fraction containing blast cells as well as fractions containing unproliferated cells manifest the same degree of suppressor capabilities. However, when density gradient separation of T cells followed by subsequent incubation with Con A was performed, fractions of proliferating cells of low density exhibited no suppression; a fraction containing high density T cells produced marked suppression, but this fraction incorporated only little thymidine in response to Con A. Thus, these studies indicate that Con A-induced suppressor T cells belong to a distinctive subpopulation which has already been programmed to express this function before exposure to Con A and that cell proliferation may not be a prerequisite for the development of such suppressor T cells.


Induction of plasma cell differentiation of human fetal lymphocytes: evidence for functional immaturity of T and B cells. Resembling the in vitro antibody response of the newborn cultures of cord blood lymphocytes stimulated with pokeweed mitogen (PWM) generated fewer plasma cells (PC) than comparable adult lymphocyte cultures and the response was almost exclusively of the IgM class. We investigated the cellular basis of this difference by preparing mixed cultures of newborn T or B lymphocytes with adult B or T cells. Substitution of adult for newborn T cells enhanced the response of newborn B cells, particularly of the IgG and IgA classes. The response of second trimester fetal spleen cells was also increased by adult T cells, although no IgA PC appeared. Conversely, adult B cells generated fewer PC particularly of the IgG and IgA classes when cultured with newborn T cells. The relatively poor IgA and IgG responses of newborn cells seems partially but not entirely due to deficiency of T cell helper function. Suppressor activity of newborn T cells was investigated by adding excess unrelated newborn or adult T cells to adult T + B cells: adult T cells improved the response whereas newborn T cells were variably suppressive. The results indicate that newborn T cells, although capable of helper function, are balanced toward suppression.


20alpha-hydroxysteroid dehydrogenase: a T lymphocyte-associated enzyme. 20alpha-Hydroxysteroid dehydrogenase (20alpha-SDH), an enzyme which reduces progesterone to 20alpha-dihydroprogesterone, was found to be associated with T lymphocytes. 20alphaSDH activity was present in spleen cells bearing theta antigen, spleen cells nonadherent to nylon wool (T lymphocyte-enriched population), and in  thymocytes. Almost no enzymatic activity was found in bone marrow cells from normal mice and in spleen cells from neonatally thymectomized or athymic nude mice. T cell mitogens (PHA and Con A), but not the B cell mitogen LPS, induced high levels of enzymatic activity 48 hr after addition to spleen cell cultures. The level of 20alphaSDH activity in lymphocytes was age dependent. At the age of 4 weeks 20alphaSDH activity in thymocytes, spleen cells, and lymph node lymphocytes was 3 to 5 times higher than at 8 and 16 weeks. Progesterone (5.0 X 10(-7) M) was found to inhibit thymocyte proliferation after exposure to mitogens, but not 20alpha-dihydroprogesterone (10(-6) M). 20alpha SDH may protect the embryonic thymocytes against high concentrations of progesterone.


In vitro immune response of human peripheral lymphocytes. I. The mechanism(s) involved in T cell helper functions in the pokeweed mitogen-induced differentiation and proliferation of B cells. Human peripheral lymphocytes (PBL) upon stimulation with PWM proliferate and differentiate to IgM- and IgG-producing cells. The PWM-induced Ig production in B cells was dependent on T cells, and cell-free supernatant (CFS) obtained from PWM-stimulated PBL or T cell-rich fraction replaced T cell helper functions. The active substance(s) in CFS were most likely derived from T cells. The kinetic studies showed that the proliferation of B cells took place in advance of the final differentiation to Ig-producing cells and that T cells or T cell product(s) had to exist at the initiation of cultures in order to give the maximum helper effect. However, the final differentiation of B cells to Ig-producing cells was not dependent on T cells. The helper effect of T cells or T cell product(s) on PWM-induced proliferation and differentiation of B cells was exerted across the MHC barrier. This may make it possible to apply this experimental system to the assessment of quantitative and/or qualitative changes in human helper T cells in several immunologic diseases.


Characterization of suppressive immunoglobulin-binding factor (IBF). I. Production of IBF by a theta-positive lymphoma (L-5178-Y). L-5178-Y, a theta-positive, Fc receptor-bearing mouse thymoma cell line spontaneously releases immunoglobulin-binding factor (IBF) upon short-term incubation in vitro. IBF produced by L-5178-Y cells is identical in its biologic activity with IBF produced by Fc receptor positive alloantigen-activated T cells. It suppresses the in vitro plaque response of mouse spleen cells to sheep erythrocytes by interfering mainly with the late phase of the generation of antibody-forming cells. Therefore, L-5178-Y thymoma affords a homogeneous source of IBF in sufficient quantities for the study of its biochemical nature and the mechanism by which it interferes with cells participating in antibody synthesis.


Select growth of human T lymphocytes in single phase semisolid culture. Selective growth and clonal proliferation of human T lymphocytes can be achieved by using a single-phase semi-solid methylcellulose system without the requirement of preincubation with lectins. Significant proliferation, however, depends upon the continued presence of Con A or PHA, but not pokeweed mitogen or lipopolysaccharide within the methylcellulose. This procedure eliminates nonspecific agglutination by lectins and allows for direct visualization of colonies and their specific removal and subsequent cloning in liquid phase. Optimal growth and production of colonies greater than 40-cell size require 3 to 9 days. Individual cells can be identified on the basis of E rosette formation and absence of surface immunoglobulin or ability to phagocytize latex particles. Moreover, proliferation is inhibited by antithymocyte but not anti-B cell sera and can be demonstrated with peripheral blood T and MOLT-4 cells, but not with B or Raji cells. Finally, colony formation is not enhanced by the presence of 2-mercaptoethanol. The clonal proliferation of human T lymphocytes has wide application in the study of both antigen recognition and lymphocyte alterations in specific diseases.


Induction of I region-restricted hapten-specific cytotoxic T lymphocytes. Murine cytotoxic T lymphocytes (CTL) reactive to TNP-conjugated syngeneic target cells do lyse to a moderate but significant extent TNP-conjugated, I region compatible but H-2K or H-2D region incompatible target cells. Antibody inhibition experiments and "cold inhibition" experiments indicate that some CTL clones recognize TNP-conjugated targets in association with syngeneic I region determinants independently of H-2K or H-2D gene products. The data suggest that besides TNP-conjugated H-2K or H-2D gene products, in principle, also TNP-conjugated I region determinants do stimulate TNP-specific CTL precursor cells and act as target antigens of TNP-specific CTL.


Characterization of antigen-binding receptors in vitro. II. Antigen-binding capacity and affinity of lymphoid receptors after immunization. We examined the kinetics and affinity of antigen binding in lymphoid populations in mice after immunization. There is increased binding capacity in lymphoid cells from animals that have undergone primary immunization. This increase would seem to be related to increased numbers of antigen-binding cells (rosette-forming cells). The serum antibody titers rise after the increasing binding capacity and numbers of BSA rosette-forming cells have increased. There is an increased amount of antigen bound per antigen-binding cell at certain times after immunization with two peaks in this capacity being demonstrable--one occurring at 4 days after immunization and the second occurring approximately 12 days after immunization and persisting for prolonged periods after that. With time, after immunization two separable peaks of increased antigen-binding cells become apparent, one very early (before Day 4) and one later (after Day 20 to 30). The affinity constants for antigen-binding cells have been measured and found to be high, and to increase with time after immunization. It appears that the heterogeneity of the affinity constants for antigen-binding cells is high early in immunity and becomes more homogeneous with time after immunization.


Effect of macrophages and antibodies on in vivo growth of Moloney sarcoma in the rat. Brown Norway and Lewis rats were challenged with a Brown Norway Moloney sarcoma tumor, MST-1, admixed with nonimmune peritoneal exudate macrophages syngeneic to the host; or admixed with nonimmune peritoneal exudate macrophages and hyperimmune anti-MST-1 antibodies. In vivo growth of MST-1 in BN and Lewis rats was inhibited by admixing Brown Norway or Lewis macrophages, respectively, with BN anti-MST-1 antibodies. The inhibiting BN antibodies were of the IgG2 class, lacking IgG2a antibodies. Brown Norway anti-MST-1 of IgG2 class without macrophages did not affect growth of MST-1. Brown Norway and Lewis anti-MST-1 antibodies of IgG2a class enhanced tumor growth, whether admixed with macrophages or not. Anti-MST-1 antibodies of IgM and IgG1 classes did not influence tumor growth. Peritoneal exudate macrophages removed from Lewis donors 8 to 10 days after inoculation of MST-1 inhibited completely growth of the challenge tumor; macrophages of Brown Norway origin were inhibitory only when harvested from hyperimmune donors, that is, 40 or more days after inoculation of MST-1. Macrophages from hyperimmune donors were specifically cytotoxic to MST-1 and did not inhibit an unrelated syngeneic BN tumor of chemical origin.


Specific binding of T lymphocytes to macrophages. II. Role of macrophage-associated antigen. Peritoneal exudate lymphocytes from immune guinea pigs that bind in vitro to autologous antigen-pulsed macrophages were allowed to proliferate for 1 week to give a population markedly enriched in antigen-specific T cells. This enriched population was then studied with regard to its binding to fresh autologous antigen-pulsed macrophages. Specific binding was not inhibited by a large excess of antigen in the media (5000-fold greater than the amount of antigen associated with the macrophages) either soluble or bound to Sepharose beads, or by coating the antigen-pulsed macrophags with antibody to the exogenous antigen, by reacting a second layer of antibody to the heterologous antibody, or by haptenating the antigen and treating the hapten-antigen macrophage complex with excess anti-hapten antibody. Results of treating antigen-pulsed macrophages with the proteolytic enzymes trypsin and pronase indicate that exogenous antigen is on the macrophage surface, but the experiments failed to prove that the removable antigen is essential for binding. The simplest interpretation of these results is that the T cell receptor is not specific for native exogenous antigen.


Receptors for IgM on certain human B lymphocytes. A receptor for IgM was demonstrated on the surface of human B lymphocytes by using a rosette technique with ox erythrocytes coated with rabbit IgM antibody (EAM). Lymphocytes forming rosettes with EAM did not bind sheep red cells, had membrane Ia-like antigens and, in some instances, surface immunoglobulin. The specificity of EAM rosettes was confirmed by inhibition experiments with purified human Ig. IgM but not IgG molecules inhibited the rosette reaction. In addition, inhibition of EAM rosettes with IgM fragments showed that the receptor has affinity for a part of the molecule located in the Fc portion. By analogy with the receptors previously found on certain human T cells, receptors for IgM were not detected on freshly isolated B cells, but were expressed after overnight culture in IgM-free media. Studies on different human lymphoid tissues showed that IgM receptors are expressed on a limited percentage of both circulating and noncirculating B cells. In addition to normal B cells, the malignant B cells of a majority of cases of chronic lymphocytic leukemia expressed the receptors for IgM.


Pharyngeal colonization with Haemophilus influenzae type b: a longitudinal study of families with a child with meningitis or epiglottitis due to H. influenzae type b. A longitudinal study of pharyngeal colonization with Haemophilus influenzae type b included 264 members of families that had a child with meningitis or epiglottitis due to this organism. It was found that (1) 52 of 67 such families contained at least one carrier of H. influenzae type b, who was usually a sibling; (2) H. influenzae type b spread slowly in 39 families colonized continuously during a six-month period, with only eight of 19 uncolonized siblings acquiring the organism during that time; (3) 18 of 30 initially colonized families contained one or more carriers after 12 months, including 30% of initially colonized siblings; (4) the highest carrier rate of H. influenzae type b occurred in recovered patients, 80% of whom were colonized after hospital discharge; (5) titers of antibody in serum were higher in colonized than in uncolonized individuals (P less than 0.001); (6) levels of antibody in colonized children were lower in those younger than two years than in older children (P less than 0.001); and (7) prolonged or heavy colonization with H. influenzae type b was not associated with unusually high titers of antibody.


Circulating capsular antigen in infant rats infected with Haemophilus influenzae type b. The kinetics of bacteremia and capsular antigenemia in infant rats infected with Haemophilus influenzae type b were measured by quantitative bacterial counts in blood and counterimmunoelectrophoresis of plasma. After intraperitoneal inoculation with 10(4) colony-forming units (cfu) of H. influenzae type b, bacteremia was detected in 100% of animals at 12 hr after inoculation (mean, 16,500 cfu/ml) and by two days exceeded 10(5) cfu/ml in most animals. Despite these high levels of bacteremia, capsular antigen was detected infrequently during the early phase of experimental infection; it was present in 20% of animals at 12 hr and in 50% at one day. Peak levels of antigen in blood occurred two to three days after inoculation and coincided with the histologic appearance of meningitis. Thereafter, the frequency of antigenemia declined and paralleled the decline in quantitative bacterial counts in blood. Since detection of antigen was dependent on the occurrence of prolonged infection, counterimmunoelectrophoresis proved to be an insensitive method for early diagnosis.