Induction of erythropoietin responsiveness in vitro by a distinct population of bone marrow cells. Bone marrow contains a small population of primitive erythroid progenitor cells which can be detected by their capacity to form large numbers of erythroid progeny in viscous cultures containing erythropoietin (EP). These cells have been termed erythroid 'burst-forming units' (BFUe). The present study demonstrates that expression of the erythroid differentiation potential of BFUe requires the presence of an activity additional to EP. This activity has been designated as BFA (burst feeder activity). It is shown that the number of BFUe detected and their apparent sensitivity to EP are directly related to the BFA concentration of the cultures. BFA was found to be associated with a population of bone marrow cells of high buoyant density and small volume, which are sensitive to irradiation. The radiation dose-effect curve provided strong evidence that bone marrow BFA is independent of cell proliferation; this was supported by showing that BFA is unaffected by in vivo treatment with hydroxyurea. The findings are compatible with a two-step regulation model for erythroid differentiation in which BFA-induced progeny of BFUe acquire sensitivity to EP.


Fine structure of the rhombencephalic tela of the bullfrog, Rana catesbeiana. The posterior rhombencephalic tela choroidea of the bullfrog was examined by electron microscopy. This membrane, the pia-ependymal roof of the caudal hindbrain, contains a large central region characterized by cuboidal ependymal cells which surround sizable microscopic apertures -- the interependymal pores. Ultrastructurally ependymal cells of this area are characterized by infrequent apical microvilli and cilia. They contain irregularly shaped nuclei and few cytoplasmic organelles that are largely apical in position. The most striking feature is an abundance of cytoplasmic filaments forming an extensive cytoskeleton. Laterally these cells are joined by numerous elaborate desmosomes. The majority of the ependymal cells have a basal lamina consisting of single, double, or triple laminae lying parallel to the basal plasma membrane. Several unusual specializations are seen at the margins of the interependymal pores. The ependymal cells have lateral cytoplasmic processes that form the actual border of each pore. These processes originate from the apical surface of the cell and partially enclose an elaborate network of basal lamina associated with the interependymal pores. These findings demonstrate microscopic apertures in the roof of the fourth ventricle in the bullfrog that are associated with an unusual form of supportive ependyma.


Stimulation of septation in mitochondria by diamide, a thiol oxidising agent. Diamide (10-(4)M), a thiol oxidizing agent, rapidly promotes septation in isolated frog liver mitochondria an also in situ in liver slices. The effect is partially inhibited by dithioerythritol. DNP does not have this effect, and it is concluded that diamide does not promote septation via an uncoupling action. The septate mitochondria have a different appearance from typical dividing mitochondria previously described; it is suggested that (1) diamide may act by favouring the fusion of the internal membranes, and (2) -SH oxidation is of importance in mitochondria in ageing and in various pathological conditions. The ways in which septa may develop in mitochondria in the orthodox and condensed configuration are discussed.


Tumor promoters and epidermal growth factor stimulate anchorage-independent growth of adenovirus-tranformed rat embryo cells. Previous studies indicated that the potent tumor promoter 12--0--tetradecanoyl-phorbol-13-acetate (TPA) enhances transformation of rat embryo cells (2 degrees RE) by a mutant of human Ad5 (H5ts125). This study examines the effect of TPA, its structural analogs and epidermal growth factor (EGF) on anchorage-independent growth of a cloned population of H5ts125-transformed 2 degrees RE cells (clone E11). Both TPA and EGF (approximately 10(-8) M) induced a 3--5 fold increase in agar cloning efficiency of E11 cells. In addition, macroscopic colonies appeared earlier and were larger and more diffuse. The TPA analogs phorbol--12,13--didecanoate (PDD) and ingenol--3,20--dibenzoate also enhanced growth in agar of E11 cells, whereas phorbol, 4 alpha PDD and 4--0--meTPA, which are inactive as tumor promoters, failed to enhance agar growth. In contrast to the results obtained with E11 cells, TPA, PDD or ingenol--3,20--bidenzoate failed to induce growth in agar of normal 2 degrees RE cells. Dexamethasone (10(-5)--10(-6) M), trans retinoic acid (10(-5)--10(-6) M) and the protease inhibitors leupeptin, antipain and elastatinol did not inhibit the ability of TPA to enhance the growth of E11 cells in agar. The TPA-enhanced anchorage independence was a stable property, since subclones of E11 cells isolated from TPA-agar plates had a higher agar cloning efficiency than the parental E11 cells when retested in the absence of TPA. This effect of TPA does not appear to reflect simple selection of a subpopulation of cells. When the parental E11 cells were first cloned in monolayer culture in the absence of TPA, all ten randomly picked clones showed enhanced growth in agar in the presence of TPA. In addition, prior growth of E11 cells in monolayer culture in the presence of TPA did not enhance their subsequent growth in agar. This system therefore provides an example in which TPA appears to enhance the acquisition of a stable cell property, and thus may be a useful model for studying mechanisms of tumor promotion and progression.


Synchronization of MEL cell commitment with cordycepin. The response of differentiating MEL cells to the nucleotide analogue cordycepin reveals a previously unrecognized aspect of the molecular events which cause commitment of these cells to terminal erythroid differentiation. Cordycepin rapidly inhibits commitment of DMSO-treated MEL cells in a dose range which does not cause cytotoxicity. Reversal of cordycepin treatment in the presence of inducer leads to a rapid and synchronous commitment of a significant proportion of cells in the culture. These results suggest that MEL cells can be blocked just prior to the point of commitment by cordycepin treatment.


Significance of the 48-hour plasma level in high-dose methotrexate regimens. Plasma methotrexate (MTX) concentrations at 48 hours were determined for 40 patients with osteosarcoma who received 256 infusions of high-dose methotrexate-citrovorum rescue (HD-MTX-CF) regimen. Five manifestations of toxicity (dermatitis, stomatitis, myelosuppression, liver dysfunction, and kidney dysfunction) were considered in the assessment of the overall toxicity. Logistic regression analysis was applied to study the effect of number of prior infusions, age, and 48-hour MTX plasma level on the risk of moderate or severe overall toxicity. Each factor had a significant effect with P less than 0.08. The predicted incidence of moderate-severe overall toxicity in the high-risk group (48-hour MTX level greater than 1.00 x 10(-6) mol/l., prior infusions greater than 10, age greater than or equal to 15 years) was 33.2% compared to only 2.4% in the "low-risk" group (48-hour MTX level less than or equal to 1.00 x 10(-6) mol/l., prior infusions less than or equal to 10, age less than 15 years). The plasma MTX determination at 48 hours postinfusion was found to be independent of both dose infused and patient's age.


alpha-Amylase of human pure pancreatic juice: effects of pancreatic disease and the occurrence of variant forms in pancreatic juice from healthy volunteers. Pure pancreatic juice (PPJ) from healthy human volunteers and from patients with pancreatic or liver disease was subjected to isoelectric focussing (IEF) and assayed for alpha-amylase activity. In PPJ from most normals, a single predominant form of amylase was found, comprising congruent to 83% of the total activity recovered, and having pIapp congruent to pH 6.8. In PPJ from six normals, variant principal forms of amylase were found at pH congruent to 6.4 or pH congruent to 7.3, in addition to the peak at pH 6.8. IEF patterns of PPJ from individuals with pancreatic or liver disease were indisquishable from patterns obtained with PPJ from the control group of healthy volunteers.


Immunological and genetic markers in a family with Hashimoto's disease. A family with Hashimoto's disease in three generations is described. Seven persons (all female) had Hashimoto's disease was the only thyroid disorder occurring in this family; no other autoimmune disease was observed. Circulating thyroid antibodies were detected in all seven subjects with overt thyroiditis in this family. Thyroid antibodies were also detected in low titres in about half of the healthy relatives. Evidence of thyroid antigen-directed cell-mediated immunity was demonstrated using the leucocyte migration inhibition test in four out of seven subejcts with thyroiditis and also in about half of the relatives without clinical thyroid disease. The relative number of thyroglobulin-binding circulating lymphocytes was elevated in six subjects with Hashimoto's disease. Again, the percentage of such cells was also increased in about half of the 'healthy' relatives. Thyroid-stimulating immunoglobulins were detected by the radioreceptor assay in three of the seven subjects with Hashimoto's disease and in four out of thirteen relatives without overt signs of thyroiditis. In conclusion, all subjects with Hashimoto's disease carried immunological markers of autoimmune thyroid disease in the circulation. In addition, most of the 'healthy' relatives were also positive for some or all of the markers sought in this study. The expression of these markers thus seems to be variable. No clear-cut conclusion could be drawn regarding the inheritance of these markers. HLA genotypes were assayed for thirty-five specificities of A, B and C loci and five of the D loci. There was no correlation between any individual antigen or HLA haplotype and overt Hashimoto's disease in this family.


Urinary free cortisol excretion in patients with hyperserotoninaemia from the carcinoid syndrome. To determine if patients with chronic hyperserotoninaemia from the carcinoid syndrome have increased basal adrenocortical function, I have determined the urinary free cortisol excretion of seventeen patients with carcinoid tumours and the carcinoid syndrome, twelve patients with carcinoid tumours without the carcinoid syndrome and seventeen normal subjects. There was no significant difference in the urinary free cortisol excretion of the patients with carcinoid tumours and the carcinoid syndrome (133 +/- 20.0 nmoles/24 h), patients with carcinoid tumours without the carcinoid syndrome (115 +/- 29 nmoles/24 h) and the normal subjects (96 +/- 9 nmoles/24 h). There was no correlation between the urinary free cortisol secretion and urinary 5-hydroxyindoleacetic acid or serum serotonin concentration in the patients with the carcinoid syndrome. There was a suggestion that patients with 5-hydroxytryptophan (5-HTP) secreting carcinoid tumours had higher urinary free cortisol excretion than patients with predominantly serotonin (5-HT) secreting carcinoid tumours. This may be due to the fact that the non-polar 5-HTP molecule penetrates the blood-brain barrier more effectively than the polar 5-HT molecule. 5-HTP is then converted to 5-HT within the brain. None of the twenty-nine patients with carcinoid tumours had clinical or laboratory evidence of the ectopic ACTH syndrome.


Chlormezanone in the treatment of insomnia due to rheumatic stiffness. An open assessment of chlormezanone in the treatment of rheumatic sleep disturbances was carried out in general practice. Sixty-one patients received 400 mg chlormezanone (2 tablets) each night for 2 weeks. Patients' nightly sleep assessment showed a steady improvement over the 2-week period, and overall effectiveness was rated as good or excellent by 75% of patients. Only 1 patient stopped treatment because of side-effects. It was concluded that chlormezanone could prove a useful alternative to current therapy for treatment sleeplessness in rheumatic patients.


A study of non-articular rheumatic disorders and their response to treatment with naproxen sodium. A group of 85 patients suffering from non-articular rheumatic disorders was studied in an open trial of naproxen sodium in general practice. Patients were assessed and then treated for 7 days with naproxen sodium at a dosage of 275 mg 3 or 4-times daily depending on the severity of pain. Patients were asked to keep a daily record of symptoms and were re-assessed by the doctor after 7 days and, in some cases, after 14 days. Pain and limitation of movement were the predominant symptoms at admission. Patients' daily records showed statistically significant reductions in pain from Day 1 and significant reductions of limitation of movement from Day 2 of the study. At the 7-day follow-up, 27 were cured, 35 improved, 19 not changed or worse, and 4 were not assessed. Thirty patients were given a second weeks' therapy and at the end of this period 24 were cured or improved. Indigestion was the most common side-effect, but only 1 patient withdrew from the trial because of this. One patient developed a rash and was withdrawn from the trial. Both patients were taking the lower dose.


Clinical and pharmacological evaluation of different preparations of oral erythromycin. The activity of erythromycin stearate in respiratory tract infections was evaluated. This study involved 269 out-patients. Of these, 76 satisfied the strict criteria for detailed bacteriological study with positive cultures of a known pathogen before treatment and bacteriological follow-up after treatment. The results of erythromycin stearate were good against the haemolytic streptococcal infections. The activity of this antibiotic was shown to be insufficient against Haemophilus influenzae. A second clinical trial with 20 patients with acute exacerbations of chronic bronchitis demonstrated quite good activity from erythromycin ethylsuccinate and no intolerance when administered in a dose of 1 g q.i.d. The administration of 1 g of the ethylsuccinate immediately after a meal gave a peak serum concentration after 60 min of 3.36 micrograms/ml. Blood levels on the fourth day of this treatment showed some accumulation, concentrations of greater than 4 micrograms/ml being found at 30, 60 and 90 min after the dose. With its good absorption, its lack of toxicity and its reputation for good diffusion into bronchial secretions, we consider that erythromycin ethylsuccinate should have a major place in the treatment of respiratory tract infections.


K-Cell activity in lamina proprial lymphocytes from the human colon. Using an enzymatic method for their isolation, it has been shown that human colonic lamina proprial lymphocytes, isolated from patients with various colonic diseases and depleted of polymorphonuclear leukocytes and macrophages, demonstrated K-cell activity in two antibody-dependent cell-mediated cytotoxicity assays in vitro. No differences were found between the activities of lamina proprial lymphocytes isolated from colonic specimens involved by Crohn's disease, chronic ulcerative colitis, or colonorectal carcinoma. K-cell activity was demonstrable using lymphoid cells isolated from the colons of patients receiving steroid therapy at the time of the therapeutic resections. These findings indicate the need for further investigation of possible roles for K-cells in local immune responses in the human bowel mucosa in inflammatory and neoplastic diseases.


Effect of sex hormones on HL-RH-degrading hypothalamic enzyme system during estrus cycle in rats. Basal activity of L-cysteine arylamidase, an LH-RH-degrading enzyme, was determined in the hypothalamus of rats at 4 h intervals throughout the 4-day estrus cycle. The activity of the enzyme system fluctuated during the four estrus stages in a circadian rhythm with maximal values in the night and lowest values at noon. The injection of increasing doses of estradiol-17 beta at various estrus stages caused a moderate stimulation of enzyme activity with no relationship to endogenous hormone levels or estrus stage. The highest activation of the hypothalamic enzyme in response to the application of progesterone occurred at such periods of the cycle when endogenous plasma progesterone is known to be low, and vice versa. When luteinizing hormone or prostaglandin E2 were injected i. v. during the various estrus stages maximal stimulation could be observed at diestrus. The LH-RH-degrading L-cystine arylamidase in the hypothalamus of the rat seems to play a modulating role in the regulation of the tonic LH release during the estrus cycle, but apparently does not influence the events governing the preovulatory LH-peak.


Labelled amino acids in plasma of patients with thyrotoxicosis and thyroid cancer after radioiodine treatment. The levels of total organically bound 131I, per cent of labelled iodoproteins, total labelled iodinated amino acids and a percentage of individual iodinated amino acids (thin-layer chromatography) were measured in serum of 84 patients with thyroid cancer and of 16 patients with thyrotoxicosis at 48 h after the administration of therapeutic dose of 131I. In thyrotoxic patients treated with therapeutic doses of 131I (2 to 39.6mCi; 74 to 1500 MBq) the findings were similar to normal subjects. In patients with thyroid cancer a significant increase of iodotyrosines was found after thyroid radioiodide ablation (100 to 200 mCi; 3.7 to 7.4 GBq). No remarkable differences were found between two groups of patients with thyroid cancer, the first one being treated with thyroid eliminating dose in attempt to activate the metastases of functionally differentiated tumour, while the second one was treated with similar doses (i. e. 100 to 200 mCi) to suppressor destroy a functionally active tumour or its metastases. Even though the hormonogenesis in tumours was hardly distinguishable from the products of its radiation damage, it was suggested that the hormonogenesis in neoplastic tissue differs from that in normal thyroid only quantitatively, being less in patients with thyroid cancer than in these with thyrotoxicosis.


The evidence of production or activation of a natriuretic factor in the liver. The natriuretic activity was measured with the aid of bioassay in blood flowing out from the brain, kidney and liver before and after the expansion of extracellular space in dogs. In another experiments in rats the volume natriuresis and the natriuretic activity in blood were measured in controls and in animals with carbon tetrachloride liver dystrophy and pharmacological stimulation (dehydrocholic acid, Ca pantothenate, cyanocobalamin) of liver functions. In dogs, the level of natriuretic activity in blood flowing out of the liver before and after the expansion of extracellular fluid volume was significantly higher than that in arterial blood. In contrast, the natriuretic activity was not found in the blood flowing out of the brain and the kidney. In rats the experimental dystrophy of the liver decreased the content of a natriuretic factor in blood plasma and the expansion of extracellular space did not produce any natriuretic effect as compared to control rats. After pharmacological stimulation of the liver by dehydrocholic acid, Ca pantothenate and cyanocobalamine the volume expansion led to a significant increase of the excretion of sodium by the kidney and to the increase of the content of a natriuretic factor in blood plasma as compared to control rats. These facts are considered to support the view that a natriuretic factor is either synthesized or activated by the liver.


Effects of gonadal steroids on hypothalamic and pituitary RNA and protein synthesis in rats. The sexual differences and the effect of castration and of a single i. p. injection of oestradiol-17 beta or testosterone on RNA and protein synthesis have been studied in the anterior pituitary, hypothalamus and cortex from rats. It was found that: 1. Sex-, castration- and tissue dependent differences were found in RNA content and in in vitro synthesis of nucleic acids and proteins. 2. Oestradiol showed a general facilitatory effect on RNA and protein synthesis in every tissue examined in 4-8th and 8-16th day after treatment respectively. This facilitation was followed by a suppression of RNA synthesis in the pituitary from castrated males and females and by that of protein synthesis in the hypothalamus from ovariectomized and in the pituitary from orchidectomized rats. 3. The first significant change in the above measures appeared simultaneously in the pituitary and hypothalamus or - under some experimental conditions - the pituitary anticipated the hypothalamus in its response to steroid treatment.


Peroxidase activity in the rat breast tissue as a marker of estrogen action. Peroxidase activity and its inducibility by estrogen in the uterine and breast tissues of Holtzman rats varied with the age of animals. Basal proxidase level of the two tissues was low in immature and high in older animals. Estrogen induced significant peroxidase activity in the uteri of rats up to 56 days of age and thereafter the uterine response was poor. Though the breast tissue peroxidase response to estrogen was significant in 20-160 days old female rats, greater inducibility being noted between 40-90 days of age. Among the sex steroids, only estrogen could induce the enzyme in the breast tissue in a dose dependent way. Peroxidase was also induced appreciably in the male breast tissue to an extent observed in age-matched females. It is suggested that this enzyme could be regarded as a diagnostic end point of estrogen action in the rat breast tissue.


19-Nortestosterone in mouse kidney.

19-Nortestosterone in mouse kidney. 19-Nortestosterone was identified as a product of testosterone conversion in vitro in male mouse kidney. After incubation of [14C]testosterone with kidney slices of castrated male mice, 64% of the radioactivity was recovered as testosterone, about 6% as epitestosterone and 2.5% was identified as 19-nortestosterone. The renotrophic effect of 19-nortestosterone was at least as high as that of testosterone. Possible role of 19-nortestosterone in mouse kidney is discussed.


Age differences in the effect of glucagon on the metabolism of glucose precursors in perfused rat liver. The effect of glucagon on the uptake and incorporation of 14C-pyruvate and 14C-alanin into glucose was studied in perfused liver of young (aged 10 days) and adult (aged 120 days) male rats. It was found that the processes studied were significantly higher (P less than 0.005) in young animals, the difference being still more remarkable after the injection of glucagon (10 microgram kg-1) to the animals (liver donors) one hour before the dissection of liver.


Inhibition of lymphocyte proliferation by polyamines requires ruminant-plasma polyamine oxidase. Spermine and spermidine in vitro are potent inhibitors of proliferation of phytohaemagglutinin-stimulated rat thymic lymphocytes, lymphoma cells and human lymphoblastic leukaemia cells, but only in media supplemented by foetal calf serum. This inhibition is shown to be due to a bovine plasma polyamine oxidase, with a high specificity for these polyamines. Spontaneously dividing lymphocytes are not subject to this inhibition. This, plus direct evidence from synchronous cultures of EB2 cells demonstrates that the inhibition is expressed in the late G1 or G1/S interface of the cell cycle. Putrescine was not an inhibitor in the presence of foetal calf serum but became so in the presence of human pregnancy serum, possibly due to the action of diamine oxidase.


In vitro immunosuppressive effect of serum from orally immunized mice. Mice were orally immunized by 4 daily intragastric (IG) intubations of 4 X 10(9) SHEEP RED BLOOD CELLS (SRBC). The serum of such mice (OS), taken two weeks after the first intubation, was compared to normal mouse serum for its suppressive effect on in vitro primary responses of normal spleen cells to SRBC. OS regularly depressed the in vitro response to less than 10% of the normal response. The suppressive effect was dose-dependent, antigen-specific and removable by absorption with SRBC. When cultures were stimulated with SRBC pre-exposed to OS, the response was also depressed. The suppression was less when the addition of OS was delayed. OS from BALB/c mice efficiently depressed in vitro responses of spleen cells from other strains. Spleen cells, taken 2 weeks after IG immunization, responded normally to in vitro challenge with SRBC. The suppressive factor had the molecular size of an IgG and could be removed by anti-Ig and anti-IgG1 but not by anti-alpha or anti-mu immunosorbents. Purified IgG1 was more suppressive than IgG2a and IgG2b despite a similar anti-SRBC antibody content.


Deficient production of anti-red cell autoantibodies by mice with an X-linked B lymphocyte defect. The production of anti-self red cell antibodies has been studied in mice bearing the CBA/N X-linked B lymphocyte defect and their phenotypically normal littermates. Defective mice produced anti-self red blood cell (RBC) antibodies at markedly reduced levels after multiple injections of rat RBC. They also failed to show significant numbers of plaque-forming cells against enzyme-treated isologous RBC. These findings are discussed in terms of the nature of the CBA/N defect and the acquisition of a complete specificity repertoire.


Frequencies of mitogen-reactive B cells in the mouse. Lipopolysaccharide-, lipoprotein- and Nocardia mitogen-reactive B cells in CBA/N mice. B cell-defective (CBA/N X BALB/c)F1 male mice have 100 times less lipopolysaccharide-reactive and 10--100 times less lipoprotein- and Nocardia mitogen-reactive B cell precursors than F1 female mice with normal B cells.


Experimental gynogenesis provides evidence of hybridogenetic reproduction in the Rana esculenta complex. The gynogenetic offspring of the hybrid frog Rana esculenta (R. ridibunda X R. lessonae) are exclusively of the ridibunda type. This is due to the premeiotic exclusion of the lessonae genome from the hybrid's germ cells.


Small ventricles detected by CT-scanning in epileptic patients. Relatively young 119 cases of generalized epilepsy and 99 controls of an identical age were selected. The bicaudate CVI was measured and the results were as follows: 1) In our cases, the mean of bicaudate CVI showed a smaller value than those of Hahn's in both epilepsy and control groups. 2) When the value of bicaudate CVI was set up below the point of 8.5 as "excessively small ventricle," the incidence rate of "excessively small ventricle" was 9% in the control group and 31% in the epilepsy group. The data showed higher incidences of small ventricles in the epilepsy group, and that the younger females have a tendency to have higher incidences of small ventricles. 3) To make clear the reason for the higher incidences of small ventricles in epilepsy, the small ventricle group and the non-small ventricle group in epilepsy were compared with each other from the points of the clinical course, EEG findings and frequency of attacks. No correlation or difference between them was found. Further investigation will be needed to make clear the etiology of small ventricles in epilepsy. However, it could be said at the present stage that not so many atrophic changes were observed in the younger patients of generalized epilepsy.


Demonstration of surface membrane immunoglobulin on L  lymphocytes by the mixed antiglobulin rosetting reaction (MARR) and by the direct antiglobulin rosetting reaction (DARR). Various preparations of human peripheral blood lymphocytes were examined for the presence of membrane-incorporated immunoglobulin (Ig) molecules using the direct antiglobulin rosetting reaction (DARR) and/or the mixed antiglobulin rosetting reaction (MARR). In an initial study, 97% of lymphocytes depleted of sheep erythrocyte (E) rosette-forming cells were found to be Ig positive by the MARR. This suggests that the majority of E rosette-negative lymphocytes are Ig-bearing B lymphocytes. Since most investigators divide E rosette-negative lymphocytes into an Ig-positive or B-cell population and an Ig-negative or null-cell population, it was decided to study further a well defined null-cell subset. Lymphocytes with membrane-labile IgG markers or L lymphocytes are reported to be E rosette negative, Ig negative by direct immunofluorescence (DIF), C3 receptor negative and Fc receptor positive. L lymphocytes were separated from DIF-positive B cells then tested for the presence of surface Ig, approximately 80% of L lymphocytes were found to be Ig positive with both the MARR and the DARR. Comparison of gamma, alpha and mu heavy chain determinants on both DIF-positive and L lymphocytes indicated that Ig isotypes are similar on both cell types. These various findings suggest that most L lymphocytes are of B-cell lineage.


Human thymus cells: a study of rosette-forming capacity with unsensitized and sensitized erythrocytes of various species. Human thymus cells from 117 donors aged 2 months to 15 years, were studied for rosette-forming capacity with unsensitized and sensitized erythrocytes of various species. Viability of the thymus cells at the completion of isolation ranged from 95 to 99%. Over 90% of the total population of thymus cells were identified as T lymphocytes by either standard Es-rosette assay utilizing a 4 degree incubation with preceding centrifugation or T--LCL assay, on the average. Gravity Es-rosette assay high affinity Es-rosette assay, stable Es-rosette assay or active Es-rosette assay for identification of subsets of T lymphocytes indicate that most of the human thymus cells were capable of forming rosettes with unsensitized sheep erythrocytes at higher degrees of temperature with or without preceding centrifugation. Approximately half of the human thymus cells were also capable of forming rosettes with unsensitized allogeneic erythrocytes. Less than 3% of the thymus cells expressed receptors for the Fc portion of IgG (T gamma suppressor cells) whereas less than 1% of thymus cells expressed receptors for the Fc portion of IgM (T mu helper cells), indicating that most of the thymus cells are non-T mu/non T gamma cells. Only 1% or less of the thymus cells were identified as B cells by the SIg assay, the Em-rosette assay or the Ia-like antigen assay. There was no significant difference of any rosette assay between males and females or among various age groups.


Maturation of mitogenic response in foetal guinea-pig. Lymphocytes obtained from thymus, spleen and heart blood of 33-day-old guinea-pig foetuses and from lymph nodes of 48-day-old foetuses onwards were stimulated by phytohaemagglutinin (PHA). concanavalin A (Con A) and dextran sulphate (DxS). The results, based on the analysis of ninety-five foetuses, indicated that the mitogenic responses of the guinea-pig T cells matured first in the thymus and then in the spleen and lymph nodes, in that order. The PHA and Con A responses in the thymus, spleen and lymph nodes and the DxS response in the spleen improved with the increasing age of the foetus. A clear improvement of the mitogenic responses did not take place in the blood. Thus, at birth the guinea-pig is immunologically mature as regards the PHA and Con A responses in the thymus, spleen and lymph nodes and the DxS response in the spleen. Regarding the maturation of PHA and Con A responses by lymphocytes from the blood, further studies are needed, since at birth both of these responses are clearly below the adult level.


On the composition of IgG anti-IgG immune complexes. The formation of different immune complexes of IgG was followed in a system comprising human IgG as antigen and rabbit IgG directed against the Fc portion of human IgG as antibody. Soluble IgG complexes were analyzed by analytical zonal centrifugation. In antigen excess, 16S complexes predominated. 16S complexes are oligomers of IgG, mainly trimers and tetramers. In decreasing antigen excess larger and larger complexes were formed. It was, however, found consistently that oligomers were always formed in the largest amounts. The largest complexes detectable by this method consisted of about twenty IgG molecules. The solubility of different complexes in polyethylene glycol was also studied. Low concentrations of polyethylene glycol preferentially precipitate large complexes. Four and six per cent polyethylene glycol precipitated all types of IgG complexes although not completely. Polyethylene glycol was seemingly not bound directly to soluble immune complexes, but caued otherwise soluble complexes to precipitate by an indirect mechanism.


Relationships among differentiated T-cell subpopulations. V. T-cell dependent mechanism of xenograft rejection and its radioresistant nature. Rat tumours grew progressively in nu/nu mice but not in nu/+ mice, normal AKR mice or sublethally irradiated AKR mice after subcutaneous inoculation. Suppression of tumour growth appeared to depend on radioresistant cytotoxic T cells which were detected in in vivo neutralization tests (Winn's test) with spleen cells of mice immunized with rat lymphocytes or tumour cells 5 days previously. Radioresistant cytotoxic activity was detected by 51Cr-release test in glass-non-adherent peritoneal exudate cells 5 days after intraperitoneal immunization with rat lymphocytes. These results suggested that immune lymphocytes might differentiate to mature cytotoxic cells at the site of direct graft rejection.


The collaboration of T-cell subsets in the mitogenic stimulation of purified B-cell subpopulations. A highly purified preparation of mouse B cells showed greatly decreased incorporation of [6-3H]-thymidine when stimulated with pokeweed Pa-1 mitogen or bacterial lipopolysaccharide compared with mouse splenic lymphocytes. This decreased stimulation was restored by the addition of purified T cells, but not macrophages. Nylon-adherent T cells exerted this helper activity only toward complement receptor-positive B cells (CR+B cells). whereas the helper activity of nylon-non-adherent T cells was effective only on complement receptor-negative B cells (CR-B cells). Since the helper activity of nylon-adherent T cells was completely abolished by the treatment with anti-Ia antiserum and complement but that of nylon-non-adherent T cells was not, it was assumed that Ia+T cells were helper cells for CR+B cells and Ia-T cells helper cells for CR-B cells. Moreover, these helper activities of both T-cell subsets were mediated by soluble factors, which were effective just before the onset of DNA synthesis of the corresponding B-cell subpopulations.


Removal of intraocular lenses.

Removal of intraocular lenses. The following points should be considered regarding removal of an IOL: 1. Be sure that it is an offending agent and not an innocent bystander before deciding to remove it. 2. An offending IOL may usually be replaced with an anterior chamber lens, with excellent results. 3. Removal of an IOL is usually a difficult procedure in less-than-satisfactory circumstances, and should be undertaken only by an experienced implant surgeon, with special attention to the delicate nature of the patient's eye and emotional feelings about this dilemma. 4. Every effort should be made to perform maneuvers in a closed system with minimal opening and folding of the cornea and minimal irrigation of the anterior chamber. 5. An offending IOL should be removed and replaced with an appropriate lens before irreversible damage has been done to the eye, if at all possible.


Activation of ribulose 1,5-bisphosphate carboxylase from Rhodopseudomonas sphaeroides: probable role of the small subunit. The activation properties of the form I and form II ribulose 1,5-bisphosphate carboxylases from Rhodopseudomonas sphaeroides were examined. Both enzymes have a requirement of Mg2+ for optimal activity. Mn2+, Ni2+, and Co2+ can also support activity of the form I enzyme, whereas only Mn2+ can substitute for Mg2+ with the form II enzyme. The effect of different preincubations on the carboxylase reaction was also examined. Both enzymes exhibited a lag when preincubated with other than Mg2+ and CO2 before assay, but the lag was much more pronounced and the rate of the reaction was slower with the form I enzyme under these conditions. Activation of the form I carboxylase By Mg2+ and CO2 occurred more rapidly than that of the form II enzyme. The results obtained with the two distinct forms of carboxylase from R. sphaeroides, as well as studies with the spinach and Rhodospirillum rubrum enzymes, thus indicate that the presence of the small subunit affects the rate of activation by Mg2+ and CO2 as well as the rate of reactivation of ribulose bisphosphate-inactivated enzyme.


Bacteriochlorophyll a-types in chromatophore and subchromatophore preparations from Rhodopseudomonas sphaeroides. Comparison of absorption and circular dichroism (CD) spectra in the near infrared region was made with chromatophore and subchromatophore preparations obtained from Rhodopseudomonas sphaeroides. The 850 nm absorption band had a positive correlation with the 850 nm and 870 nm CD bands. The 800 nm and 870 nm absorption bands seemed not to correlate with any CD bands. Lipid contents in chromatophores and subchromatophores were measured. Lipids in membranes seemed to contribute to the appearance of the 870 nm absorption band, but not to that of the 800 nm and 850 nm absorption bands. The time courses of absorbance changes were compared at 800, 850, and 870 nm in detergent-treated chromatophores. Relative changes of absorbances differed from one another. The present results suggest that the three absorption bands are due to three different bacteriochlorophyll a-types and the 850 nm absorption band originates from exciton-coupling of bacteriochlorophyll a.


Hydrophobic-ionic chromatography. Its application to purification of porcine pancreas enzymes. 1. All the porcine pancreas enzymes tested, regardless of their pI's were adsorbed on Amberlite CG-50 (a weakly acidic cation exchange resin) at pH 4, where the ion-exchange group (carboxyl group) is not dissociated. The adsorption is hardly influenced by ionic strength. 2. At pH 4, the adsorbed enzymes were partially eluted by organic solvents such as 50% propanol. 3. The adsorbed enzymes were effectively eluted by increasing the pH from 4 to 6. Trypsin (pI 10.5) was eluted before carboxypeptidase A (pI 4.5 AND 5.3) WITH 0.5 M acetate buffer, whereas the former enzyme was eluted after the latter enzyme with 0.2 M 3,3-dimethyl glutarate buffer. However, with either buffer, the elution order of enzymes was not always the same as the order of the pI's. 4. By a single Amberlite CG-50 column chromatography of porcine pancreas extracts, kallikrein, carboxypeptidase B, deoxyribonuclease, carboxypeptidase A, and trypsin were purified 100-fold, 16-fmately 13%. The purification procedures included treatment with protamine, ammonium sulfate fractionation, treatment with acid, DE-32 cellulose column chromatography, gel filtration on Sephadex G-100, preparative polyacrylamide gel electrophoresis, and affinity chromatography on 5' AMP-Sepharose 4B. The last procedure, affinity chromatography on 5' AMP-Sepharose 4B, was useful for the removal of other dehydrogenases. The enzyme which was homogeneous, as shown by polyacrylamide gel electrophoresis, had a molecular weight of about 92,000. The optimum pH was at 10.0 and isoelectric point at 5.2. The enzyme accepted both L-fucose and D-arabinose as substrate, but was specific for NAD+ as coenzyme. Km values were 0.15 mM, 1.4 mM, and 0.07 mM for L-fucose, D-arabinose, and NAD+, respectively. A single enzyme catalyzed the oxidation of L-fucose and D-arabinose, which had the same configurations of hydroxyl groups from C-2 to C-4. The reaction products obtained with L-fucose as substrate were L-fucono-lactone and L-fuconic acid. The L-fucono-lactone was an immediate product of oxidation and was hydrolyzed to L-fuconic acid spontaneously. This reaction was irreversible. Therefore, it is likely that L-fucose dehydrogenase is involved in the initial step of the catabolic pathway of L-fucose in rabbit liver.


Cultivation of Pneumocystis carinii with WI-38 cells. Pneumocystis carinii has been successfully cultured with WI-38 human embryonic lung fibroblasts. Inoculum was obtained from infected lungs of cortisone-treated Sprague-Dawley rats. Trophozoites reached peak numbers between days 4 and 8 and grew in two subcultures, but then proliferation ceased. If primary cultures were harvested and new medium were added, a second and sometimes a third harvest could be obtained. Cell monolayers were not destroyed. After growth of the initial inoculum had ceased, monolayers supported growth of a new inoculum. Harvested organisms had few contaminating tissue culture cells. Additional studies of growth requirements are needed. Although cultures cannot be used to diagnose P. carinii infection, cultured organisms should be useful for studies of biology and pathogenesis and for the development of immunodiagnostic techniques.


Meningitis due to Haemophilus influenzae type e biotype 4. Haemophilus influenzae type e biotype 4 was isolated from the cerebrospinal fluid of a 16-month-old child with meningitis. This is the first isolation, from a case of meningitis, of this organism that has been biotyped.


Acquired von Willebrand's syndrome with IgM inhibitor against von Willebrand's factor. This report describes a patient without evident underlying disease, in whom an acquired von Willebrand's syndrome was discovered before surgery. Coagulation abnormalities included a borderline bleeding time, a low retention of platelets on glass beads, decreased levels of factor VIII procoagulant activity (VIIIAHF), factor VIII-related antigen (VIIIAg), and ristocetin-induced agglutination cofactor (VIIIVWF). After cryoprecipitate infusion the patient did not have the expected rise and there was no secondary increment in VIIIAHF. The patient was treated with prednisone for three weeks without significant improvement in the laboratory findings. Spontaneous resolution was observed long after this therapy. The haemostatic abnormalities were attributable to the presence of an inhibitor directed against VIIIVWF. The inhibitor was found in the IgM fraction. Its autoimmune nature is probable although we failed to demonstrate any inhibitory effect of Fab obtained from the patient's purified IgM. Despite the lack of inhibitory effect against VIIIAHF and VIIIAg, the low levels of all three activities of the factor VIII complex could be explained by the short half-life of immune complexes between factor VIII and the inhibitor.


Changes in T and B blood lymphocytes after splenectomy. The blood lymphocytes of 37 splenectomised patients were analysed by means of T and B lymphocyte surface markers. Sixteen patients had had a splenectomy for non-haematological and 21 for haematological reasons. The results show that 15 had normal numbers of T and B cells; decreased T cells were found in two patients, raised B cells in seven, raised T and B cells in eight, and raised T cells in five patients. Increased numbers of 'null' cells were observed in some patients, especially in those with raised B cells. Follow-up studies indicate that raised levels of T and B cells can be established by one to three months post-splenectomy and may persist, although in some patients the cells fall to normal levels. The lymphocyte proliferative response to phytohaemagglutinin and Concanavalin A in vitro was normal in eight out of nine patients with raised T cells and was depressed in one patient, possibly due to an intrinsic cell defect.


Target role of Langerhans cells in mycosis fungoides: transmission and immuno-electron microscopic studies. Langerhans cells of the epidermis are "special" macrophages, as indicated by their expression of Fc and C3 receptors and Ia antigen. Functionally, they can replace macrophages in presentation of antigens to T cells and in the MLR. Ultrastructural and immunologic studies were carried out on epidermal samples of mycosis fungoides. As in contact allergy, apposition of lymphocytes to Langerhans cells was noted. Destruction of Langerhans cells appeared to act as the focus for the development of Pautrier microabscesses. This destruction was accompanied by ingress of histocytic cells, probably related to the stem cell population for Langerhans cells. The target cell role of Langerhans cells appears to be similar in contact allergy and mycosis fungoides.


A statistical evaluation of the binding of ferritin-conjugated lectins to the surface of rat cells. Topographical variations as a function of lectin concentration and cell type. Statistical procedures were utilized to evaluate the concentration dependence of labeling by ferritin-conjugated lectins on four different rat cells: hepatocytes, normal thymocytes, Friend virus-induced rat tumor cells and feline sarcoma virus-induced rat sarcoma cells. Labeling by ferritin conjugates of concanavalin A, wheat germ agglutinin and Ricinus communis agglutinins I and II was quantitated by counting the number of ferritin granules on 600 Angstrom membrane segments. Relationships between the arithmetic means and variances for sample populations from each cell and ferritin-lectin combination were used to define four types of topographical distributions: uniform/ordered, uniform/random, random and clustered. It was found that the distribution and/or density of surface-bound lectin was concentration-dependent for all four ferritin-lectins. The nature of this dependency was complex and varied with both lectin and cell type.


Indirect 125I-labeled protein A assay for monoclonal antibodies to cell surface antigens. An assay for detection of monoclonal hybridoma antibodies against cell surface antigens is described. Samples of spent medium from the hybridoma cultures are incubated in microtest wells with cells, either as adherent monolayers or in suspension. Antibodies bound to surface antigens are detected by successive incubations with rabbit anti-immunoglobulin serum and 125I-labeled protein A from Staphylococcus aureus, followed by autoradiography of the microtest plate or scintillation counting of the individual wells. Particular advantages of this assay for screening hybridomas are: (1) commerically available reagents are used, (2) antibodies of any species and of any immunoglobulin class or subclass can be detected, and (3) large numbers of samples can be screened rapidly and inexpensively. We have used the assay to select hybridomas producing monoclonal antibodies to surface antigens of human melanomas and mouse sarcomas.


Changes in corticotrophin releasing factor of the stalk median eminence in rats with various cuts around the medial basal hypothalamus. The activity of corticotrophin releasing factor (CRF) in extracts of the stalk median eminence (SME) complex proper (average protein content, 30.6 micrograms) of male rats was assayed by monolayer cultures of anterior pituitary cells using the release of immunoreactive ACTH. Extracts which were equivalent to 0.025 SME of control rats usually had detectable CRF activity, while there was no detectable activity in extracts of 0.4 SME equiv. taken 8 days after complete surgical isolation of the medial basal hypothalamus (MBH). The activity of CRF in extracts from rats with an anterolateral cut around the MBH was at least ten times less than that in the control rats. One day after placing an anterolateral cut around the MBH the ACTH releasing activity of the SME was not significantly different from that of the control animals but activity decreased significantly 3 days after the operation and was at least ten times less than in the control animals on day 7 after the operation. It is suggested that most of the CRF activity of the SME is contained in nerve fibres entering the neurohaemal region from outside the MBH and that transection of these fibres produced the fall in CRF content of the SME in rats with partial or total surgical isolation of the MBH.


Cross protection among togaviruses in nude mice and littermates. After immunization with Sindbis virus, T-cell deficient nude mice, compared to normal littermates, were equally protected against challenge with Sindbis virus. However, the nude mice showed about one-tenth the protection observed with normal littermates after challenge with Semliki Forest virus at a dose of 100 LD50. This consistent with our previous interpretation that sensitized T-cell populations play a major role in cross protection between the two togaviruses. The remaining low level of specific cross protection in nude mice (detectable only at a challenge dose of 10 LD50) could not be attributed to an anamnestic response of neutralizing antibody to the challenge virus or to an effective antibody-dependent, complement-mediated cytolysis of infected cells in vivo. Other possible compensatory mechanisms to explain the low level of specific cross protection in nude mice are discussed.


Lymphocyte subpopulations in multiple sclerosis: serial studies and clinical correlations. Total lymphocyte counts and T and SIg+ cell numbers in peripheral blood were determined in 74 healthy controls and 44 MS patients. Twenty-five patients were studied in relapse and at two intervals after ACTH. Nineteen had not relapsed for 6 months but most had progressed clinically. MS patients showed significantly lower total lymphocyte counts which were not correlated with disease activity. Subpopulation analyses showed the MS patients to have significantly lower T cell numbers and significantly elevated SIg+ cell numbers and percentages. Although T cell numbers were low in all phases, both T and SIg+ cell abnormalities were maximal in progressive disease or in the recovery phase after relapse and a significant fall in T cell percentage and rise in SIg+ cell number was confirmed in serial studies of 16 patients followed from relapse to recovery. SIg+ cell numbers correlated negatively with relapse rate and correlated positively with the ratio of disability to number of relapses. The incidence in MS patients of HLA A3 was significantly increased and of HLA B12 and HLA BW40 significantly decreased but no correlation was seen between HLA phenotype and T, SIg+ or total lymphocyte counts. It is suggested that the peripheral blood SIg+ cell abnormalities and some of the T cell abnormalities are secondary to the pathological process, perhaps due to antigenic stimulation and particularly in non-relapsing progressive disease. Their possible role in influencing recovery or progression is discussed.


Idiotype anti-idiotype complexes in cerebrospinal fluids of multiple sclerosis patients. The involvement of idiotype-anti-idiotype complexes in multiple sclerosis was approached by ultracentrifuge studies. Specimens of the immunoglobulin G fraction obtained from the cerebrospinal fluids of multiple sclerosis patients which contained distinct oligoclonal bands, were subjected to analytical ultracentrifugation. None of these samples revealed the presence of any detectable amount of oligomeric immunoglobulin G, namely dimer or trimer. By comparison with control mixtures containing known amounts of dimeric immunoglobulin, it was evaluated that the cerebrospinal fluid samples contain less than 5% dimer. These findings indicate that the spinal fluids of multiple sclerosis patients do not contain idiotype-anti-idiotype complexes to an extent that would account for the oligoclonal immunoglobulin bands as representing complementary idiotypes and anti-idiotypes.


Large-field color naming of dichromats with rods bleached. A color-naming method was used to examine the large-field red/green discriminations of dichromats screened with standard tests. The stimulus was a 12 degrees annular field with the central 4 degrees removed, flashed for 300 ms. Four wavelengths were equated in brightness for each observer at two retinal illuminance levels, approximately 10 and 100 trolands. The stimuli were then presented in random order and the observer was asked to name each, using one of four color terms. The entire experiment was done with the observers dark-adapted and also with the rods bleached. For all four deuteranopes and two of four protanopes, color names were very significantly related to both illuminance and wavelength in both adaptation conditions. The relationship between name and wavelength was similar to that of a normal trichromat, but the performance of a dichromat was very poor by comparison. Performance was generally somewhat better in the dark than with the rods bleached. However, the result in the bleached condition is consistent with recent evidence that at least some observers who are classified as dichromats with standard small-field screening procedures actually have a weak residual third cone mechanism.


Estimation of the density of sodium entry sites in frog skin epithelium from the uptake of [3H]benzamil. 1. The inhibition of short circuit current in frog skin by benzamil (N-benzyl-amidino-3,5-diamino-6-chloropyrazine carboxamide) was investigated. When skins were bathed on both sides by Ringer solution (pH 7.6) the affinity was 5 x 10(7) M-1. When the sodium concentration was reduced to 1.1 mM and the pH adjusted to 6.5 the affinity increased to 8.5 x 10(8) M-1. 2. A method is described for measuring uptake of [3H]benzamil into the mucosal (outer) surface of pieces of isolated epithelium, 0.95 cm2 in area, under open circuit conditions. 3. The relation of [3H]benzamil uptake at the mucosal surface to its concentration was measured in solutions containing 1.1 mM-sodium and adjusted to pH 6.5. Uptake could be resolved into a linear component (10.2 f-mole nM-1) and a saturable component (21.5 f-mole cm-2) with a half saturating concentration of 1 nM. 4. In the presence of amiloride (1 microM) or unlabelled benzamil (1 microM) uptake was linear with concentration, and was, respectively, 9.2 f-mole nM-1 and 8.8 f-mole nM-1. When the pH was reduced to 3.5 uptake was again linear but reduced to 3.3 f-mole nM-1. 5. The identity of the saturable component of [3H]benzamil uptake to sodium entry sites is discussed. The results suggest a sodium entry site density of around 130 micron-2 of mucosal surface.


Uptake of [3H]benzamil at different sodium concentrations. Inferences regarding the regulation of sodium permeability. 1. The effect of benzamil on short-circuit current in frog skin was measured at different external sodium concentrations. A linear relationship exists between the concentration of benzamil reducing short-circuit current by 50% and the external sodium concentration, indicative of some form of competitive antagonism between sodium and benzamil. 2. Uptake of [3H]benzamil into isolated frog skin epithelium and whole skin (0.95 cm2 pieces) was measured at different external sodium concentrations. With a sodium concentration of 111 mM in the external medium the uptake of [3H]benzamil is linear with concentration. Uptake amounted to 8.8 f-mole nM-1, a value similar to the linear component of the uptake measured at low (1.1 mM) sodium concentration. 3. Using a variety of other conditions the maximal number of specific binding sites for [3H]benzamil was calculated from displaceable binding and the fractional occupancy, the latter being derived from the inhibition of short-circuit current. This approach gave similar binding site densities to those reported previously at low sodium concentrations. 4. The reduction in specific [3H]benzamil uptake at high sodium may result from two mechanisms, competition of sodium with the ligand for an external binding site and a reduction in the site density as the intracellular sodium concentration increases. 5. It is concluded that the saturation of sodium transport which occurs at high sodium concentration is likely a consequence of the reduced availability of entry sites, rather than saturation of the uptake process.


The action of salicylate ions on the frog node of Ranvier. 1. The effects of salicylate on the membrane currents in the frog node of Ranvier were investigated. 2. External salicylate slows the falling phase of the action potential, with little effect on the action potential amplitude. 3. External salicylate has no effect on the leak current. 4. Most of the actions of external salicylate can be attributed to a simple incorporation of negative charge into the membrane surface. The h infinity, tau h, tau m and m infinity curves for the sodium current are shifted to more negative potentials, as are the n infinity and k infinity curves for the potassium current. Potassium ion accumulation prevented analysis of the action of salicylate on the time constant of the potassium current kinetics. 5. In addition to the h infinity and tau h curves being shifted to more negative potentials, the shapes of the curves are also changed by salicylate. These shape changes cannot be explained by conventional homogeneous surface charge theory. Possible explanations for these changes are discussed. 6. Internal salicylate has similar effects to external salicylate: the gating variable curves for the sodium current are shifted in the negative direction on the voltage axis, rather than in the positive direction expected if negative charge were added to the inner surface of the nodal membrane. This may be due to salicylate crossing the membrane and binding preferentially to a receptor at the external surface, or might be due to a rise in intracellular calcium concentration following inhibition of oxidative phosphorylation.


Primary health care in the New Hebrides. The New Hebrides is a small Melanesian country in the South-West Pacific whose doctors are almost entirely recruited from France and Great Britain, the two countries which jointly administer the territory. This paper describes briefly the difficulties of providing primary health care for a fairly primitive island society.


Antitumor and antiinflammatory agents: N-benzoyl-protected cyanomethyl esters of amino acids. A series of N-protected cyanomethyl esters of various amino acids was synthesized and tested for antineoplastic and antiinflammatory activity in rodents. Utilizing the L-phenylalanine cyanomethyl ester and varying the N-protecting moiety demonstrated that the N-tosyl and the N-Cbz analogues were the most active against Ehrlich ascites cell proliferation. The N-(carbobenzyloxy)- and N-benzoyl-L-phenylalanine cyanomethyl esters were the most active against carrageenan-induced inflammation. In the N-benzoyl series of cyanomethyl esters, L-alanine, DL-valine, and L-leucine amino acid analogues were the most active against Ehrlich ascites cell proliferation. The glycine and L-alanine analogues possessed the best inhibitor activity in the antiinflammatory screen. The cyanomethyl esters also demonstrated immunosuppressive activity and the ability to suppress the writhing reflex which is associated with inflammatory pain. However, no antipyretic or narcotic analgesic activity was demonstrated by these agents.


Comparative effectiveness of five analgesics for the pain of rheumatoid synovitis. Utilizing 3 different methods for analyzing the results, a single dose double-blind crossover study of the effectiveness of 5 analgesics and placebo against the pain of rheumatoid synovitis in 30 subjects showed all active drugs superior to placebo by 1 analysis; aspirin, codeine and acetaminophen superior to both placebo and propoxyphene by a second; and aspirin alone superior to placebo and propoxyphene by a third. More side effects were reported from pentazocine than from other agents.


Patient compliance: a novel method of testing non-steroidal antiinflammatory analgesics in rheumatoid arthritis. Patient compliance has been used as a measure of efficacy of non-steroidal antiinflammatory drugs (NSAID) in the treatment of patients with rheumatoid arthritis. A series of studies on various cohorts of patients using different drugs and different prescribing methods has been conducted. The studies have confirmed the absence of long-term placebo response. Indomethacin emerges as the most effective drug regardless of the mode of prescription, but it is clear from the results that the level of compliance varies with the mode of prescription.


Infectious arthritis due to Hemophilus influenzae. A healthy 51-year-old male developed multiarticular infectious arthritis due to Hemophilus influenzae, a rare cause of infectious arthritis in adults. Previous case reports are reviewed. Predisposing factors include chronic illness, underlying joint disease, joint trauma, and respiratory infection. H. influenzae is frequently misidentified on Gram stain, being mistaken for gonococci or pneumococci. Infections due to H. influenzae may occur in normal adults. Aspects of immunity are discussed.


Interaction between caffeine and adenosine on the membrane current and tension component in the bullfrog atrial muscle. Antagonistic action of caffeine and adenosine were studied at the membrane current and tension component levels in myocardium of the bullfrog atrium, mainly under voltage clamp using the double sucrose-gap method. Caffeine (1-5 mM) produced marked augmentation of the phasic and tonic tensions as well as slow inward current (Is). At higher concentrations it elicited an increase of the delayed outward current (Ix). The augmentation of Is was principally due to increase of the limiting conductance (gs), while the activation and inactivation variables (dinfinity, finfinity) were not changed significantly. The dose-tension response curve for caffeine, which appeared sigmoidal, was notably lowered in the presence of adenosine (1-3 mM), indicating a competitive type of inhibition. Adenosine, in the presence of caffeine, exerted a narcotic-like action by inhibiting all of the membrane currents (INaf, Is, Ix and background inward current, Ib) and tension components (ICa-dependent and independent tensions). The inhibition of Is was due to a decrease of gs and no appreciable change was observed in dinfinity and finfinity. These results suggest that adenosine has a strong stabilizing action on the myocardium, especially when the heart muscle activity is accelerated by increased cyclic AMP.


Contractility of the frog ventricular myocardium in sodium-free lithium solution. The effects of Na-free Li solution on the electrical and mechanical activities were examined in a bullfrog's ventricular strip. The action potential showed a triphasic change during Na-free perfusion: the initial decrease in the overshoot and duration, the secondary restoration and the gradual decrease in the membrane potential. The twitch contraction also changed in a triphasic manner: the initial rapid decrease with a transient development of contracture and the secondary slow decrease followed by a partial recovery. A negative staircase, which was accompanied by full-sized twitch contractions, was induced by lowering the stimulation rate in this medium, indicating an involvement of some intracellular Ca releasing sites for activating the well-maintained contractile proteins. When CaCl2 was omitted during Na-free Li perfusion the twitch decreased only slightly down to 71.8% whereas in Na-containing medium it decreased to 11.9%. The sensitivity of twitch to the external Ca was thus greatly diminished in the Na-free condition. A fairly large contracture could be elicited by high K solution in Ca-free Li solution even after a prolonged Ca-free perfusion. Modification of the action potential by passing a hyperpolarizing current did not affect the twitch occurring in the Li solution, in contrast to its marked effect on the muscle perfused with normal Ringer. From these results it is concluded that the twitch is activated mainly by Ca released from some intracellular site in the Na-free Li solution. The inhibitory effect of Mn on this contraction was discussed in relation to its intracellular action.


Response patterns of olfactory bulb neurons to stimulation of distilled water and odorous solutions. Responses of 50 olfactory bulb neurons of the bullfrog to odorous solutions of ethyl n-butyrate were composed mainly of facilitation, inhibition and no response, whereas those of the same neurons to water were mainly of facilitation and delayed facilitation. The responses to water tended to continue longer than those to the odorous solutions.


[Modern methods in localization of pheochromocytomas (author's transl)]. In six patients with adrenal pheochromocytoma the tumors were localized by ultrasonography, phlebography of the adrenal glands and by estimation of plasma catecholamines selectively obtained from the vena cava and the adrenal gland veins. All tumors were localized by selective catecholamine estimation, five by ultrasonography, and four by phlebography. The smallest pheochromocytoma of 1.5 g weight was only localized by selective catecholamine estimation but not by ultrasonography or phlebography. This tumor, however, had been visualized by computed tomography. To avoid diagnostic errors by selective catecholamine estimation, it is important to withdraw blood from the adrenal gland veins prior to the injection of any radiographic contrast media, since this may result in an extremely enhanced secretion of catecholamines from the adrenal medulla.


A new approach in the relief of pain of leprous neuritis. Transcutaneous Nerve Stimulation (TNS) has been known since the last several decades to relieve pain in many conditions. For the first time it was used in the treatment of the severe and agonizing pain caused by leprous neuritis with highly beneficial results without producing any side effects. This study was made on 40 patients, and in the majority of the cases there was total relief of pain with one application of a few hours duration. This encouraging result has led the authors to the conclusion that TNS could be a useful tool in a hospital where leprosy patients are treated.


Immune suppression by cultured lymphocyte supernates of tumor-bearing hosts. A nonspecific immunosuppressive factor produced by lymphocytes of tumor-bearing hosts has been demonstrated. A modified Jerne plaque was used to quantify IgM antibody production of sheep erythrocyte-immune Swiss mouse lymphocytes (IL). Significant suppression of IgM antibody production was found when IL were treated with cultured lymphocyte supernates of 4198 fibrosarcoma-bearing mice. The suppressive activity was found to be significant six days prior to any visible tumor development, and, through the use of glass and nylon wool columns, appears to be produced by T cells. Similar immunosuppressive activity was found in 22 cancer patients tested as above, compared either to 16 patients who previously had had solid tumors but are now without clinical disease or to 19 normal controls.


Effects of age on cellular immune responses in BALB/cJ mice: increase in antibody-dependent T lymphocyte mediated cytotoxicity. Two in vitro cell-mediated immune (CMI) responses of splenic lymphocytes of BALB/cJ mice of various ages (1.25, 2.5, 4, 10.5 and 30 months) were examined. These were the antibody-dependent lymphocyte-mediated cytotoxicity (ADLMC) responses, mediated by the K cell subclass to T lymphocytes, and phytohemagglutinin (PHA) induced mitogenesis, mediated by the theta positive T lymphocytes. ADLMC responses increased with age, culminated at 10.5 months, then slowly declined. Lymphocytes of young mice (2.5 or 4 months of age) were significantly more responsive to PHA than lymphocytes of mature (10.5 months) or aged adults (30 months) (p less than 0.01), indicating progressive suppression of this type of CMI response with age, as observed previously. The difference in time of maturation and extent of suppression of ADLMC and PHA responses was not influenced by changes in numbers of specific effector cells, since percentages of these did not change with age (as measured by immunofluorescence and rosetting techniques). The increase with age of ADLMC responses may be due to deregulation of earlier suppressor activity which allows the K cell to be fully responsive, but other mechanisms may be involved.


[Diseases associated with chronic bronchopathies and plasma alpha-1-antitrypsin deficiency (author's transl)]. Two groups of patients were selected according to their plasma levels of alpha-1-antitrypsin deficiency from among 58 patients with a chronic bronchopathy. Group I had normal plasma levels of alpha-1-antytrypsin; group II had plasma values lower than normal. The pathologic conditions associated with chronic bronchopathies were studied in both groups and so were the gasometric characteristics of the same. Chronic bronchopathies in subjects belonging to group II showed a clear tendency to present normal levels of pCO2 and hydrogen ions possibly related to a greater bronchial impairment in these patients. Independently of the genetic characteristics of plasma alpha-1-antitrypsin deficiency, its general levels are the real indication of its possible etiopathogenic action. Patients with recurrent plasma alpha-1-antitrypsin deficiency, its general levels are the real indication of its possible etiopathogenic action. Patients with recurrent plasma alpha-1-antitrypsin deficiency are more susceptible to bacterial infections, liver cirrhosis, diabetes, and allergic states. All this would be related to the protective effect of this protein fraction, and its reduction according to the most recognized theories would decrease the resistance of hepatic and pancreatic cells.


[Late postpartum hemorrhage].

[Late postpartum hemorrhage]. In the presented material the frequency of postpartum haemorrhage is 0.74%. In the study sixty-nine histologically confirmed cases were considered. At histological examination three main factors were found--subinvolution of the placental site, retained placental tissue, and inflammation, each of them responsible for about one third of all cases. The possible influence of such factors as parity, time of delivery, frequency of caesarian section, frequency of intrauterine exploration, manual extraction of the placenta, and induction and stimulation of the delivery on late postpartum haemorrhage is discussed.


The nutritional regulation of T lymphocyte function. Prostaglandin (PG) E1 plays a major role in the regulation of thymus development and T lymphocyte function and the evidence for this is reviewed. The production of PGE1 is dependent on nutritional factors with linoleic acid, gamma-linolenic acid, pyridoxine, zinc and vitamin C playing key roles. Inadequate intake of any one of these will lead to inadequate PGE1 formation and defective T lymphocyte function. Megadoses of any one are likely to be only minimally effective in the absence of adequate intakes of the others. By careful attention to diet it should be possible to activate T lymphocyte function in the large number of diseases including rheumatoid arthritis, various auto-immune diseases, multiple sclerosis, and cancer in which such function is defective. It is possible that T lymphocytes may require both endogenous and exogenous PGE1 in order to function adequately. It is therefore of particular interest that many cancer cells and virally infected cells are unable to make PGE1 because they cannot convert linoleic acid to gamma-linolenic acid. The direct provision of gamma-linolenic or dihomo-gammalinolenic acids in these situations is worthy of full investigation.


Response to therapy in acute myeloid leukaemia: an explanation of two unrelated findings. Androgen therapy prolongs complete remission in acute myeloid leukaemia (AML). (1,2) In vitro studies of clone formation by myeloid leukaemia cells in semi-solid agar cultures have revealed a relationship between culture findings and response to chemotherapy. (3) These seemingly unrelated findings may be explained by the hypothesis proposed here which relates clone size in culture and sensitivity to regulators of granulopoiesis with level of differentiation in the granulocyte-monocyte pathway. This hypothesis not only suggests other means of therapy which may be equally or more effective while lacking the side effects of androgen therapy, but also predicts which patients are most likely to benefit from such therapy.


The pattern of human lymph node involvement by the non-Hodgkin lymphomas of B-cell lineage. A total of 93 lymph nodes from 24 patients with a primary malignant non-Hodgkin lymphoma of B-cell lineage was assessed histologically with special reference to the presence or absence of neoplastic B-cells in the lymph node cortex and medulla (B-cell areas) as well as in the paracortical (T-cell area) area, and in the lumen of the post-capillary venules (PVC). The B-cell origin of the lymphomas was verified by using an indirect immunoperoxidase technique for the demonstration of the cell surface immunoglobulins. The B-cell areas of the node were involved in all the lymph nodes studied, and the T-cell areas in 95.7 per cent of the nodes. Neoplastic cells were present inside the PCV in 80.6 per cent of the nodes. Furthermore, 15 lymph nodes were seen where the paracortical area was only partially involved despite the total replacement of the B-cell areas by the neoplastic cells. The results suggest that the nodal involvement by the B-cell lymphomas starts from the B-cell areas, a fact that could be used as an aid in the diagnosis of these lymphomas as well as in the study of the dynamics of B-lymphocytes in general.


[Scintigraphic and EEG diagnosis of posterior cranial fossa tumors in children]. The purpose of this work was to assess the diagnostic value of scintigraphy and EEG in posterior fossa tumours. The analysis covered 22 children aged from 10 months to 13 years treated surgically in the period from 1975 through 1978 for expanding intracranial lesions of this location. Before the operation in all cases positive ventriculography, brain scintigraphy and EEG were done. In 21 cases ventriculograms were abnormal and scintigrams demonstrated in all these cases presence of pathological foci in the posterior fossa. This method helped in accurate localization of 11 out of 12 tumours of cerebellar hemispheres and 9 of 10 posterior fossa tumours situated near the midline. EEG investigation demonstrated in 7 cases of cerebellar hemisphere tumours presence of asymmetric slow bioelectric activity in the posterior part of the brain on the side corresponding to the tumour. In EEG investigations in cases of midline posterior fossa tumours in most children symmetrical bursts of slow rhythms prevailed in posterior leads showing the characteristics of the so called á distance rhythms.


[Preliminary evaluation of epidural electric stimulation of the spinal cord]. Since 1974 clinical experiments have been conducted at the Rehabilitation Centre in Konstancin near Warsaw on the effects of electrostimulating on the damaged spinal cord. As yet stimulating electrodes were implanted in 44 cases of spinal cord injury. In the present report groups of patients with total and incomplete cervical cord injury and complete injury of the thoracic spinal cord were compared. The patients were treated by surgical decompression with simultaneous implantation of stimulation electrodes in contact with the spinal cord. The control group comprised patients operated upon in the same time period, for similar injuries, who had no stimulators implanted. The terapeutic effect was better in the stimulated patients in relation to the non-stimulated ones (this was true especially of patients with injuries to the cervical spinal cord), with a greater number of neurological improvements as well as with a better quality of these improvements. The comparison confirmed also a favourable effect of spinal cord stimulation on the development of bladder automatism which was achieved in this material twice as rapidly as in non-stimulated patients. The author thinks that these data justify further investigations along these lines and suggest that electrostimulation  will extend the possibilities of treatment of spinal cord injuries.


[Gastrointestinal bleeding in cases of ruptured cerebral aneurysms (author's transl)]. Among the 1,000 cases of direct surgery on cerebral aneurysms, two cases showed clear signs of preoperative and 19 cases showed postoperative gastrointestinal bleeding. Here we have made a clinical analysis of various aspects of the 19 cases in which the bleeding developed postoperatively. (1) Gastrointestinal bleeding was most frequent postoperatively in cases of VBA aneurysms (4.3%) and AcomA aneurysms (2.9%) and less common in MCA and ACA aneurysm cases. (2) Gastrointestinal bleeding was most frequently seen in those cases operated on between the third and seventh days from the last hemorrhage attack (9.0%) and was more common in those cases with a relatively poor preoperative grade. (3) The development of such bleeding in cases with a good preoperative grade was due to problems with the surgical operation in most cases, although the influence of vasospasm must not be ignored. The development of bleeding in cases with a poor preoperative grade is thought to be due primarily to vasospasm and transitory brain damage to the hypothalamus and the orbital portion of the anterior lobe due to hematoma caused by aneurysm rupture. (4) First the location of gastrointestinal bleeding should be determined endoscopically and, if hemostasis is not possible even after attempted coagulation, then the desirability of surgery should be determined in an early period and abdominal surgery performed.


Acute hyperesthesia after spinal trauma. Four cases of acute hyperesthesia after spinal trauma are presented. An attempt is made to characterize the natural history of this syndrome. In the three cases with truncal sensory aberration, hyperesthetic zones developed only anteriorly. All four patients demonstrated spontaneous relief of pain within 2 weeks after injury. The pain was modified by transcutaneous stimulation in one case and by l-dopa in another case. It is suggested that the "lesion" responsible for the hyperesthetic response is partial damage to dorsal root fibers at a point medial to the dorsal root ganglion.


Ribonucleoprotein antibodies, polyarthritis and metastatic squamous cell carcinoma of the bronchus. A 62-year-old woman presented with seronegative polyarthritis four months before she was found to have squamous cell carcinoma of the bronchus with widespread metastases. Unexpectedly, high titre speckled antinuclear antibodies with antibody to ribonucleoprotein were found.


Temperature experiments on nerve and muscle membranes of frogs. Indications for a phase transition. The influence of temperature changes in the range of 25 degrees C to -6 degrees C on the time constants of Na activation (tau m) and inactivation (tau h) was studied in twitch muscle fibers and the node of Ranvier under voltage-clamp conditions. Arrhenius plots of tau m and tau h exhibit a change in activation enthalpy at temperatures below 10 degrees C. Cooling and subsequent heating induce a hysteresis in the temperature dependence of tau m and tau h; Ni2+ and UO22+ increase the hysteresis width. With fast temperature changes the gating kinetics relax to their new values more slowly than the temperature change. Hence, temperature must be changed more slowly than 5 degrees C/min if an additional apparent hysteresis due simply to this relaxation is to be avoided. The data are explained by the hypothesis of a phase transition in the membrane lipids. This conception is favoured over a temperature-induced change in protein conformation, since the neutral local anaesthetic benzocaine shows use-dependent block as if low temperature restricted the access of the drug through the lipid phase to its receptor.


Contractile responses to direct stimulation of frog slow muscle fibres before and after denervation. 1. The contractile responses of single slow muscle fibres were investigated under isometric conditions. The fibres were isolated from normal and denervated iliofibularis muscle of Rana temporaria. Direct stimulation was achieved by external application of electrical pulses of depolarizing solutions (40 mM K and acetylcholine 10(-6) g/ml). 2. Upon electrical stimulation normal slow fibres developed contractions whose amplitude increased steadily with the strength of the pulses. These contractions did not exceed 0.22 kg/cm2, and their rates of rise and fall were approximately 1/6 of those observed in twitch fibres. During pulses of several 100 ms duration the tension continued to rise slowly until the end of the pulses. Application of 40 mM K or acetylcholine resulted in contractures which reached maximum values of 1.71 and 1.87 kg/cm2, respectively, after less than 1 min; little relaxation occurred during the following minutes. 3. The responses of slow fibres denervated for 9--76 days differed in several respects from those of normal slow fibres. a) The contractions elicited by electrical stimulation became faster but their amplitude decreased; the relationship between tension and pulse strength remained, however, essentially unchanged. b) Small and short contractions could be observed during and at the end of stimulating current pulses. Their amplitude was independent of the amplitude and duration of the stimuli; they were blocked by tetrodotoxin and must be attributed to action potentials. c) The ability to develop and maintain tension was reduced. Maximum values of K- and acetylcholine contractures were generally only 50% of those observed in normal slow fibres. Maintenance of tension was markedly reduced in some fibres, less so in others. 4. It is concluded that incorporation of Na channels into its membrane does not transform the slow fibre into a type of fibre which resembles a twitch fibre.


Effects of cerebellectomy on the cat's vertical vestibuloocular reflex. The effects of cerebellectomy on the semicircular canal evoked responses recorded from individual vertical and oblique eye muscles were studied in cats anesthetized with Ketamine. The phase lag relative angular acceleration of the electromyographic response was consistently smaller over the frequency range tested (0.02--1.0 Hz) in cerebellectomized than in intact animals. This finding indicates that the time constant of the central, neural integrator was shifted towards smaller values by the lesion. This was also suggested when the vertical eye movements generated by rotation about the pitch axis were recorded in the fully alert animal. In addition, the EOG data show that the phase of the VOR in the low frequency range was not altered following cerebellectomy suggesting that the macular influences were still present. When the EMG responses of the superior oblique (SO) and superior rectus (SR) muscles were studied in their presumed main mode, i.e. roll-canal and pitch-canal, respectively, no difference was noted in hemicerebellectomized animals when compared to intact control animals. However, when SO and SR were studied in pitch-canal and roll-canal rotation, respectively, dramatic changes in the response pattern were noted in lesioned animals.


[Persisting deficiency of cell mediated immunity in Hodgkin's disease in complete remission (author's transl)]. There is increasing evidence in the literature for persistent deficiency in cell-mediated immunity (CMI) in Hodgkin's diseases during apparent remissions after discontinuation of the treatment. Patients were followed for 6 months to 2 years after all treatments were stopped. There was a high percentage (41.6) of subjects with skin anergy to seven recall antigens, a highly significant (p less than 0,0001) decrease, as compared with controls, in total and active E rosettes independently from the number of lymphocytes, and a highly significant diminution of T-lymphocyte in vitro reactivity to various doses of phyto haemagglutinine (PHA). There was no difference between the patients tested between 6 months and 1 year and those tested more than 2 years after treatment. No correlation between skin tests and active rosettes was found in this series. Finally, the CMI deficiency is some what different in patients on remission and in untreated patients.


Hunger induced changes in the noradrenaline and dopamine contents in various nuclei of the limbic system in rats. Teh effect of 48 hr food deprivation on noradrenaline (NA) and dopamine (DA) content in the dorsomedial, ventromedial and arcuate hypothalamic nuclei, in the anterior and posterior part of medial forebrain bundle, in the medial preoptic region, nucleus of diagonal band (septum), and in the central, medial and basal nuclei of the amygdaloid complex was investigated by radioenzymatic assay. It was found that starvation resulted in decreased NA and DA levels in arcuate and ventromedial nuclei, and increased DA content in the posterior medial forebrain bundle. A statistically insignificant increase of DA in the central amygdaloid nucl. was also observed.


Head twitches produced by serotonergic drugs and opiates after lesion of the mesostriatal serotonergic system of the rat. Electrolytical lesions of the dorsal raphe nucleus in the rat did not change frequency of head twitches produced by codeine and apocodeine. The action of serotonergic drugs was affected differently: the lesion depressed the frequency of head twitch episodes produced by 5-hydroxytryptophan (5-HTP), LSD and quipazine, but did not change the frequency of head twitches produced by 5-methoxytryptamine. On the other hand, the lesions protected against high mortality produced by combined treatment with tranylcypromine and 5-methoxytryptamine. There was a good correlation between the extent of lesion, measured by the depression of the level of prosencephalic serotonin, and the depression of head twitch frequency produced by 5-hydroxytryptophan and quipazine, while the correlation was not significant for the action of LSD. It is concluded that presynaptic serotonergic structures belonging to the mesostriatal serotonergic system are necessary for appearing of head twitches after treatment with 5-HTP, LSD and quipazine, and therefore these compounds have a presynaptic action in this test.


[Effect of long-term administration of testosterone on the activity of T- and B-lymphocytes in male rats]. Oil testosterone propionate solution was administered to Wistar rats for 14 days in a dose of 2 mg. Some rats were immunized with sheep red cells on the 10th day of the hormone administration. The animals were sacrificed on the 5th day of the immune response. The hormone proved to suppress considerably the blast lymphocyte response of the peripheral blood to PHA, but produced no influence in laconos stimulation. A sharp increase in the antibody forming cell count was noted in the spleen, whereas the rosette forming cell count had a tendency to reduction.


Antibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus. Patients with systemic lupus erythematosus often possess antibodies against two nuclear antigens called Sm and RNP (ribonucleoprotein). We have established the molecular identity of these antigens by analyzing immune precipitates of nuclear extracts from mouse Ehrlich ascites cells labeled with (32)P and (35)S. Anti-Sm serum selectively precipitates six small nuclear RNA molecules (snRNAs); anti-RNP serum reacts with only two of these; and a third serum, characterized as mostly anti-RNP, precipitates a subset of three snRNA bands. Three of the six RNAs are identified by fingerprint analysis as the previously characterized and highly abundant nucleoplasmic snRNA species U1a (171 nucleotides), U1b, and U2 (196 nucleotides). The other three RNAs (U4, U5, and U6) likewise are uridine rich and contain modified nucleotides, but they are smaller, with lengths of about 145, 120, and 95 residues, respectively. Each of the six snRNAs is complexed with and apparently antigenic by virtue of association with specific proteins. All three sera precipitate an identical complement of seven different polypeptides ranging in molecular weight from 12,000 to 35,000; these proteins are abundant in nuclear extracts, but are neither histones nor the major polypeptides comprising the 30S heterogeneous nuclear RNP particles of mammalian nuclei. Our data argue that each of the six snRNAs exists in a separate small nuclear ribonucleoprotein (snRNP) complex with a total molecular weight of about 175,000. We find that human antisera also precipitate snRNAs from a wide range of vertebrate species and from arthropods. We discuss the antigenic snRNPs in relation to the published literature on snRNAs and nuclear RNPs and consider possible functions of snRNPs in nuclear processes.


Human lymphocyte antigens: production of a monoclonal antibody that defines functional thymus-derived lymphocyte subsets. A monoclonal mouse antibody (3A1) that specifically bound to 65% of human peripheral blood (PB) thymus-derived (T) cells but did not bind to complement receptor-positive PB bone marrow-derived (B) cells, polymorphonuclear leukocytes, or human erythrocytes has been produced. The 3AI antibody was synthesized by a stable cloned lymphocyte hybrid cell line. This lymphocyte hybrid line (3AI) was derived from fusion of P3 X 63/Ag8 myeloma cells and spleen cells from BALB/c mice immunized with HSB-2 cells, a human T cell line. The 3A1 lymphocyte hybrid line produced mouse ascites fluid containing 3A1 antibody in saturating titers of up to 1:25,600. Purified PB T cells that carried the 3A1 antigen incorporated tritiated thymidine maximally in response to phytohemagglutinin and concanavalin A stimulation, whereas purified PB T cells that lacked the 3A1 antigen responded suboptimally to phytohemagglutinin and minimally to concanavalin A. Thus, the 3A1 antibody can be easily used to study the role of 3A1-positive and negative T cell subsets in the regulation of normal and abnormal human immune responses.


The biosynthetic precursor of epidermal growth factor and the mechanism of its processing. The biosynthesis of epidermal growth factor (EGF) was studied in mouse submaxillary glands incubated with L-[35S]cystine. EGF and EGF-like proteins were isolated from the gland homogenates by immunoprecipitation with anti-EGF antiserum. The major species appearing after short labeling periods is significantly larger (Mr, 9000) than EGF. The label in the Mr 9000 species plateaus after 1 hr whereas tha in EGF continuously increases. When glands are chased with unlabeled L-cystine after a brief period of labeling, the Mr 9000 peak decreases and a corresponding amount of label appears in EGF. The Mr 9000 species was isolated from boiled homogenates in which it accounts for approximately 1% of the total EGF content. It contains five of the six chymotryptic peptides of EGF and a sixth peptide which is a modified form of the COOH-terminal chymotryptic peptide of EGF. Of the arginyl esteropeptidases, gamma subunit of 7S nerve growth factor, beta-endopeptidase, trypsin, and EGF-binding protein, only the latter converts the isolated Mr 9000 species to EGF. The extrapeptide material released in the conversion comes from the COOH terminus of the Mr 9000 species. These results suggest that the Mr 9000 species is a biosynthetic precursor of EGF and that the EGF-binding protein is the specific intracellular cleaving enzyme that converts the precursor to EGF. In the process, the stable high molecular weight complex of EGF is formed.


Endothelial plasmalemmal vesicles as elements in a system of branching invaginations from the cell surface. In electron microscopy studies of the endothelial vesicles in frog mesenteric capillaries, an accidental observation was made concerning vesicular organization. When tannic acid was added to already fixed tissue, the mordant reached apparently free vesicles in the cytoplasm under conditions in which vesicular movement was excluded and in which the impermeability of the cell membranes was preserved. This indicates a spatial continuity between the vesicles and the cell exterior. It is proposed that cytoplasmic vesicles in endothelial cells are elements of branching, permanent or semipermanent invaginations of the plasmalemma.


A human thymus-leukemia antigen defined by hybridoma monoclonal antibodies. A series of mouse hybridomas producing monoclonal antibodies against human acute lymphocytic leukemia (ALL) cells was generated and screened for tumor specificity. Among 1200 primary cultures, 60 produced an antibody that could distinguish between the immunizing leukemia cells and an isologous B lymphoblastoid cell line. Of these, two produced an antibody that detects an antigen expressed preferentially on ALL cells and on a subpopulation of normal cells found in the cortex of the thymus. Other normal human lymphoid cells from lymph nodes, spleen, bone marrow, and peripheral blood express only low levels of this antigen. High levels of this "thymus-leukemia" antigen were found on T-ALL cells, T-ALL-derived cell lines, and some "null" ALL cells. By contrast, B-cell leukemias, B lymphoblastoid cell lines, and normal and malignant myeloid cells contain either low or undetectable amounts of this antigen. The thymus-leukemia antigen has been isolated from the membranes of leukemia cells by detergent solubilization and subsequent immunoprecipitation with the monoclonal antibody. Preliminary biochemical characterization shows the antigen to be associated with a polypeptide of Mr approximately 28,000.


Patterned acquisition of the antibody repertoire: diversity of the hemagglutinin-specific B-cell repertoire in neonatal BALB/c mice. The B-cell response of 12- to 14-day old BALB/c mice to the hemagglutinin molecule of influenzae virus A/PR/8/34(H0N1) has been examined with monoclonal antibodies obtained by the splenic focus technique. An analysis of the specificity of these antibodies with a panel of heterologous viruses indicates that the antibody repertoire is highly restricted at an intermediate stage in postnatal development of the immune system. In toto, only 10 distinct reactivity patterns have been observed in an analysis of 72 antibodies derived from 28 donors. This contrasts with a substantially more diverse repertoire present in nonimmune and immune adult populations. The neonatal antibody specificities do not appear to be a random sampling of adult specificities, because several clonotypes (as defined by reactivity pattern) frequently found in neonates are rare or absent in adults. Most importantly, the vast majority of adult clonotypes are absent from the neonatal repertoire. These findings indicate that, at a developmental stage when the B-cell repertoire contains at least 10(6) clonotypes, the repertoire of genetically identical individuals is shared. This is consistent with a diversification process that is highly patterned and genetically determined. Furthermore, because 12- to 14-day-old neonates exhibit a diversified but definable hemagglutinin-specific B-cell repertoire, this experimental system should enable precise analyses of genetic and environmental influences on repertoire expression.


Differential effects of colchicine and cytochalasins on the shedding of murine B cell membrane IgM and IgD. Lactoperoxidase (EC 1.11.1.7) catalyzed cell surface radioiodination was employed to monitor  the fate of murine B cell membrane (mem) IgM and IgD on radiolabeled cells in short-term culture. Both mem-IgM and mem-IgD were shed from the cell surface with biphasic kinetics. The rapid phase of mem-IgD shedding was somewhat slower (half-time = 12 hr) than that of mem-IgM shedding (half-time = 7-8 hr). The effect of temperature, colchicine, and cytochalasin on the shedding of the two membrane immunoglobulin isotypes was determined. The shedding of mem-IgD was more energy dependent than that of mem-IgM and was sensitive to colchicine but not cytochalasin. Conversely, the shedding of mem-IgM was sensitive to cytochalasin but not colchicine. The results suggest that the mechanisms of shedding of mem-IgM and mem-IgD are qualitatively distinct and may be regulated by microfilaments or microtubules, respectively.


Thymic control of proliferation of T cell precursors in bone marrow. The activity of T lymphocyte precursors (pre-T cells) in the bone marrow of mice was measured by the concanavalin A response synergy assay. Pre-T cell levels were low in marrow of neonatally thymectomized mice and could be restored to control values by treatment in vivo with an extract of mouse thymus. Levels of activity were also low in aging mice and again could be restored by thymic extract treatment. The most profound fall with aging was in the proliferating pre-T cell compartment as detected by tritiated thymidine suicide; and this compartment was restored by thymic extract treatment. Irradiation to the thymus, with the bone marrow shielded, caused a fall in resting pre-T cells in the bone marrow and a concomitant rise in proliferating cells. These results are consistent with a model of control of pre-T cell maturation in which the thymus senses the number of developing lymphocytes within it and responds to a fall in this number by increasing production of hormone. The hormone acts on resting pre-T cells in the marrow, stimulating some of them to proliferate, leave the bone marrow, and repopulate the thymus.


Is trisomy cause or consequence of murine T cell leukemia development? Studies on Robertsonian translocation mice. Trypsin-Giemsa banding studies on T cell leukemias induced in Robertsonian translocation mice by dimethylbenz[a]anthracene and Moloney leukemia virus show a trisomy of chromosome 15 even in cases in which chromosome 15 has undergone centromeric fusion with chromosomes 1, 5, or 6. These results suggest that the duplication of gene(s) located on chromosome 15 is of critical importance for murine T cell leukemia development.