Secretion of melanophore-stimulating hormone (MSH) in long-term cultures of pituitary neurointermediate lobes. Neurointermediate lobes from pituitaries of the frog, Rana berlandieri forreri (Rana pipiens, sensu lato), were maintained in organ culture in media with and without serum for up to six months. The cultured tissues were examined periodically by light microscopy and transmission electron microscopy and by bioassay of the melanophore-stimulating hormone (MSH) secreted and present in the culture media. Light-microscopic observations revealed a high degree of preservation of the pars intermedia at four weeks with isolated areas of some glands maintaining histological integrity for the entire six months. Similarly, at the ultrastructural level the cells appeared morphologically intact and to be actively synthesizing and secreting hormone. Bioassays showed the glands to be continuously secreting MSH; however, larger yields of hormone were obtained in media lacking serum. No significant ultrastructural differences between cells grown in the presence or absence of serum were detected. The difference in concentration of MSH between the two groups therefore apparently results from enzymatic degradation of the hormone by the serum. Organ culture of the vertebrate neurointermediate lobe may provide a unique method for the production of large quantities of MSH and for the study of other melanotropic and opiate peptides as they may be synthesized and secreted by the pars intermedia.


DNA-binding antibodies and hepatitis B markers in acute and chronic liver disease. A Farr technique has been used to assay antibodies to double-stranded DNA in the serum of patients with acute and chronic liver disease and carriers of HBsAg from the United Kingdom and Iraq. These antibodies were found in all groups from both countries. The highest levels were found in chronic active hepatitis and cirrhosis. In the Iraqi patients there was a strongly positive correlation between DNA-binding antibody levels and the presence of hepatitis B markers but not with disease activity. In the patients from the United Kingdom there was little correlation with disease activity and none with autoantibodies. Ninety-five per cent of asymptomatic carriers of HBsAG had elevated DNA-binding antibodies. It is suggested that hepatitis B-specific DNA might be one trigger to DNA antibody formation, though in liver disease a variety of factors are clearly operative.


The immune-dependence of schistosomicidal chemotherapy: relative lack of efficacy of an antimonial in Schistosoma mansoni-infected mice deprived of their T-cells and the demonstration of drug-antiserum synergy. When T-cell deprived CBA mice, infected with Schistosoma mansoni, were treated orally with potassium antimony tartrate, the reduction in size of their worm burdens was less than in similarly treated, immunologically-intact animals. The defect in deprived mice could be restored by the administration of serum obtained from S. mansoni-infected normal mice simultaneously with the drug, but by a different route. A serum component, probably immunoglobulin, obtained from rabbits which had been injected with an extract from S. mansoni adult worms was also found to act synergystically with the antimonial in the chemotherapeutic eradication of S. mansoni worms from immunologically intact mice.


T- and B-lymphocyte subpopulations in the tropical splenomegaly syndrome (TSS). Lymphocyte subpopulations in thirty-nine patients with the tropical splenomegaly syndrome (TSS) were studied using the E-rosette, EAC rosette and direct immunofluorescent staining techniques. In the peripheral blood a decrease in the percentage of T-lymphocytes was found which was accompanied by an increase in the percentage and number of B-lymphocytes. These abnormalities reverted towards normal on treatment. Splenic aspirates contained an increased proportion of T-lymphocytes. The relative T-cell lymphoenia in TSS is probably due to specific trapping or non-specific sequestration of lymphocytes in the liver and splen. B-cell lymphocytosis is probably due to persistent antigenic or mitogenic stimulation by malaria.


Rabbit antiserum to human thymocyte membranes: specificity for normal and malignant T-lymphocytes. An antiserum was obtained by immunization of rabbits with human thymocyte membrane fractions. After appropriate absorptions, the antiserum was shown to detect specifically a population of T-cells. When tested by complement-mediated cytotoxicity the antiserum lysed 95% of thymocytes, 65% of normal PBL and 45% of tonsillar lymphocytes. It was also cytotoxic for three different T-cell lines (MOLT-4, CCRF-CEM and CCRF-HSB-2). When peripheral or tonsillar lymphocytes were separated into populations enriched in B- and T-cells, the percentage of cells lysed by the antiserum correlated well with the proportion of E-rosetting cells. Treatment of PBL with the antiserum and complement resulted in an increase of SmIg-positive B-cells in the residual cell fraction, which could no longer form E-rosettes. Treatment of PBL with the antiserum alone completely inhibited the E-rosette formation. The cytotoxic index on PBL from patients with various lymphoid disorders always correlated with the proportion of T-cells as assessed by E-rosette formation. Finally, the absorptive capacity of thymocytes for the antiserum was ten times higher as compared to that of PBL or tonsil cells.


B-cell activation of peripheral blood lymphocytes from patients with chronic lymphatic leukaemia. The mitogenic response of blood lymphocytes from twelve patients with B-cell CLL and seven healthy controls was investigated. All patients were untreated and had a typical B-cell dominance in their peripheral blood. The patients demonstrated a scattered response pattern with a higher variance than in the control group. We demonstrated examples of stimulation of DNA-synthesis by polyclonal B-cell activators (PBA) as well as inhibition. The variable pattern between the patients might result from a malignant transformation of cells originating from different lymphocyte subpopulations in different patients. Thus PBA studies of lymphocytes from CLL patients might be a valuable tool in characterizing further this clinically variable disease at the cellular level.


Avidity of anti-native DNA antibody and glomerular immune complex localization in lupus nephritis. Ten patients with clinically active SLE and biopsy-proven renal involvement were studied. Antigen binding capacity (ABC) and avidity of anti-nDNA antibodies were measured, using 125I-nDNA, by the ammonium sulfate test of Minden and Farr. The glomerular localization of immune complexes was determined by morphological studies. Mean avidity in 7 patients with mesangial and subendothelial deposits was 0.387 +/- 0.022 (mean +/- SE), and in those with additional subepithelial deposits was 0.247 +/- 0.036 (P less than 0.01). When measured serially, the avidity was found to alter slowly either during treatment or spontaneously. Follow-up histology in 2 cases showed morphologic transitions; in one the transition from the initial mesangial proliferative type to the membranous form was associated with a shift of the avidity from a high level to a low one. There was no significant relation between the avidity and ABC (r = 0.235, P greater than 0.1), suggesting that avidity and the quantitative ABC of antibodies in immune complexes may not necessarily behave in parallel. Thus, the avidity measurement is useful in understanding the immunological events which underlie various clinicopathological features of SLE.


The effect of quipazine, a serotonin receptor agonist, on serum corticosterone concentration in rats. Quipazine, a serotonin receptor agonist, increased serum corticosterone within 30 min after its i.p. injection (at 10 mg/kg) into rats; the effect persisted at 1 and 2 hrs, but not at 4 hrs. Elevation of serum corticosterone did not occur with a 1.25 mg/kg dose of quipazine but was dose-related over a 2.5-20 mg/kg dose range. The effect of quipazine was completely prevented by methergoline, a serotonin receptor antagonist. The effect of quipazine was present in rats pretreated with 5,7-dihydroxytryptamine to destroy serotonin nerves and was not enhanced (as was the effect of L-5-hydroxytryptophan) by fluoxetine pretreatment. These data are compatible with the idea that quipazine increases serum corticosterone as a consequence of direct stimulation of serotonin receptors in brain.


The amino-acid sequence of troponin C from frog skeletal muscle. The primary structure of the troponin C from skeletal muscle of the frog Rana esculenta has been determined. The amino acid sequence was deduced from amino acid determinations of peptides obtained after cleavage with cyanogen bromide. Overlapping peptides were isolated from tryptic digests of performic-acid-oxidized troponin C and phthalylated performic-acid-oxidized troponin C. All overlaps have been determined except for the Arg-Ile sequence at position 103--104, which has been obtained by comparison with homologous troponins C. Frog troponin C consists of one polypeptide chain containing 152 amino acids. The calculated molecular weight is 18299. There is a single cysteine residue at position 101 and a single tyrosine residue at position 112. No histidine or tryptophan residues are present. The amino-terminal amino acid is N-acetylated. The homology of frog troponin C with other skeletal and cardiac troponin C is briefly discussed.


The hatching rhythm of Polystoma integerrimum, a monogenean from the frog Rana temporaria. 1. Eggs of Polystoma integerrimum incubated in alternating periods of light and darkness at a constant temperature hatch in the light. 2. Eggs incubated and hatched in continuous light (10 degrees or 18 degrees C) hatch without a rhythm. Eggs incubated and hatched in continuous darkness at 18 degrees C hatch without any circadian rhythm but fail to hatch at 10 degees C until 1 month later than expected. 3. At both 10 degrees and 18 degrees C the hatching rhythm persists in conditions of continuous light if, during incubation, the eggs were exposed to alternating light and darkness. At 18 degrees C the rhythm appears to persist in total darkness following incubation in alternating light and darkness. At 10 degrees C hatching is blocked. 4. At 10 degrees C if eggs are incubated in alternating periods of light and darkness and then, when hatching begins, these periods are reversed, the hatching rhythm becomes synchronous with the new lighting regime. This may be due to the effect of low temperature. 5. Hatching during the day may have survival value of P. integerrimum since the host tadpoles congregate at this time around the edges of the lakes, the region where the parasite eggs are laid.


Helix pomatia A hemagglutinin: selectivity of binding to lymphocyte surface glycoproteins on T cells and certain B cells. Human and mouse lymphocytes were surface-labeled by lactoperoxidase-catalyzed iodination, or by galactose oxidase oxidation followed by reduction with tritiated sodium borohydride. The labeled cells were lysed with Nonidet P-40. Proteins binding to Helix pomatia A hemagglutinin (HP) were isolated by affinity chromatography on HP-Sepharose and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. A major cell surface glycoprotein (apparent mol. wt. 150 000, using reducing conditions) on human lymphocytes was responsible for almost all binding of HP. This protein was present on normal and malignant thymus-derived lymphocytes, e.g. thymocytes, blood T cells and T leukemia cell lines. It was also found on chronic lymphocytic leukemia cells, one null cell leukemia line, one unidentified leukemia line, one lymphoblastoid cell line of B origin and on one stem cell lymphoma line. In contrast, this protein was not found on various B cells at different steps of differentiation, e.g. four B lymphoma lines or one myeloma line. It was also absent from a histiocytic leukemia line. However, two of the four B lymphoma lines and the myeloma line had another HP-binding surface glycoprotein (mol. wt. 200 000) instead of the 150 000 protein. Studies of mouse lymphocytes similarly showed that thymus-derived lymphocytes (normal and malignant) but not normal adult B cells expressed a major HP-binding surface glycoprotein of apparent mol. wt. 130 000 (reducing conditions).


A new surface marker on mouse natural killer cells: receptors for Helix pomatia A hemagglutinin. Natural killer (NK) cells in normal mouse spleen have so far been found to lack conventional cell surface markers. Recently, neuraminidase treatment has been shown to uncover receptors for the A hemagglutinin of the snail Helix pomatia (HP) on populations (preferentially T cells) of human blood lymphocytes. We have used HP coupled to Sepharose in columns to fractionate mouse spleen cells. Fractions consisted of passed cells (fraction I) and cells eluted with the competitive hapten N-acetyl-D-galactosamine at either 0.1 mg/ml (fraction II) or 1.0 mg/ml (fraction III), NK activity and percentage of T and B cells in each fraction were determined. B cells were found enriched in fraction I. T lymphocytes, among them alloreactive cytotoxic T cells (CTL), were eluted in fraction III. However, NK activity of normal CBA spleens was highest in fraction II, fraction III was somewhat enriched, whereas fraction I was depleted of lytic activity. Thus, while NK cells possess HP receptors, their binding properties seem to differ from those of CTL. HP receptors represent the first simple and reliable marker detected on NK cells and might be useful for purification of this cell type.


Quantitative comparison of K cell potential in human T and null cells. Using a short-term quantitative assay of the antibody-dependent cell-mediated cytotoxicity (ADCC) of human peripheral blood lymphocytes towards 51Cr-labeled mouse lymphoma cells, it is possible to estimate the relative involvement of various lymphocyte populations in the overall ADCC potential displayed by unfractioned lymphocytes. Lymphocytes were separated by means of an anti-F(ab')2 column combined with sedimentation of E rosette-forming cells or EA rosette-forming cells, and a discontinuous serum albumin gradient, and were characterized by several surface markers. ADCC results were expressed as the cytotoxic capacity of 10(6) cells of each population (lytic units/10(6) cells) as well as the cytotoxic potential of each population by taking into account their relative numerical sizes. Although the null cells had the highest cytotoxic capacity, highly purified T cells also disclosed a cytotoxic capacity which, because of the large numerical excess of T cells over null cells, could account for closely equivalent cytotoxic potentials of these two populations. ADCC was entirely abolished by further removal of Fc receptor-bearing-cells, in all the subsets studied. Our data underline the role that the small proportion of Fc receptor-carrying T cells might play in ADCC.


Water compartments in the myelinated nerve. III. Pulsed NMR results. 3 experimentally distinct transverse relaxation components of the water in frog sciatic nerve are obtained by Carr-Purcell-Meiboom-Gill technique. The relative weights of these components: approximately 29%; approximately 50%; approximately 21% fit well with water compartments in this tissue as revealed by previous methods.


Chemically mediated cell-to-cell contractile activation in isolated frog atrial cardiac cells. A dying single frog atrial cardiac cell liberates an unknown substance which diffuses away from the dying cell and activates contractile activity in other isolated intact single cardiac cells within the vicinity of the dying cell.


Porphobilinogen-accumulation by a porphyrin auxotrophic strain of Bacillus subtilis. The amount of porphobilinogen accumulated by a Bacillus subtilis hemC mutant strain is dependent on time and exogenic aminolevulinic acid addition. hemC mutant seems to be suitable for porphobilinogen production on a large scale.


Inhibition of vitamin A action in rat bone cultures by inhibitors of RNA and protein synthesis. Using explanted humeri of late fetal rats, retinoic acid induced a dose- and time-dependent regression of cartilage. The retinoic acid-induced release of proteoglycan into the medium was blocked by inhibitors of RNA and protein synthesis, suggesting its dependence upon continuous RNA and protein synthesis.


Role of catechol structure in the inhibitory effects of D-isomers of amino acids on the decarboxylation of L-5-hydroxytryptophan. D-3,4-dihydroxyphenylalanine, D-threo-3,4-dihydroxyphenylserine and pyrocatechol competitively inhibited aromatic L-amino acid decarboxylase, whereas no inhibition occurred with l-norepinephrine, l-epinephrine, dopamine or some other amino acids without catechol side chains.


Tubuloreticular structures in chicken bursa Fabricii lymphocytes. Tubuloreticular structures were observed in bursal lymphocytes of a 12-day-old male chicken. These structures are often observed in mammalian lymphocytes and endothelial cells. Their presence in bursal lymphocytes confirm their association with B-lymphocytes.


Epidermotropic lymphomas. Introductory immunological remarks. The immune system is reviewed. Its ontogeny, differentiation and localization are discussed. Although cellular immunity mainly depends on T cells (thymus-derived) and humoral immunity depends on B cells (bone-marrow-derived), there are many interactions between these two systems. The role of T cells, in the production of antibodies, appears to be mainly regulatory. Various subpopulations of T cells are mentioned. The interactions between B cells and macrophages are described, as well as the markers of human T and B cells.


[Quantitative analysis of cerebral electrograms following administration of biogenic amines]. The influence of acetylcholine, norepinephrine and serotonin on the biopotentials of the cortical and subcortical-stem formations of the brain was studied in cats during a chronic experiment with monopolar electrodes. The accumulation of acetylcholine, norepinephrine and serotonin determines the formation of various profiles representing spectral characteristics of the brain electrograms, which, apparently, is conditioned by a temporal specificity of the action produced by the neuromediators on the post-synaptic membranes.


[Introduction to the modern classifications of the non-Hodgkin malignant lymphomas (author's transl)]. The authors present the historical evolution of the concepts of non-Hodgkin malignant lymphomas which lead to the new functional classifications of Lukes and of Kiel. These classifications are based on the discoveries of modern immunology and are relying on the marking techniques for the identification of the T, B and U lymphocytes. Arguments are presented why these modern nomenclatures should progressively replace the Rappaport classification which preceded the major developments of modern immunology. The Kiel nomenclature is briefly described.


Cancer immunotherapy.

Cancer immunotherapy. This paper briefly reviews some mechanisms of tumor immunity and the principles of cancer immunotherapy. Cellular and humoral immunity can both influence tumor cells. Most of the cells belonging to the immune system can act on neoplastic cells. T cells can kill them, macrophages inhibit their growth, and K cells through their Fc receptor also destroy antibody-coated tumor cells. Cancer patients have usually depressed cellular immune functions. The goal of immunotherapy is to amplify the immune reactions in order to destroy the tumor cells. The modalities of immunotherapy are described. They may become important as adjuvant therapy. Immunotherapy has already been successfully in skin cancers, lung cancer and acute myeloblastic leukemia.


T-lymphocyte dependency of B-lymphocyte blastogenic response to phytomitogens. Human peripheral blood T and B lymphocytes were separated by a method based on the stable rosette formation of T lymphocytes with neuraminidase-treated sheep erythrocytes, followed by centrifugation over a Ficoll-Hypaque gradient. Monocytes were isolated from the T-depleted B lymphocyte preparation by allowing the monocytes to ingest iron particles and by subsequent centrifugation over a Ficoll-Hypaque gradient. The T lymphocytes responded extremely well to PHA and very well to PWM, while the B lymphocytes were unresponsive to either PHA or PWM. However, when the B lymphocytes were cultured together with irradiated autologous or allogeneic T lymphocytes (1:1, 1:2 or 1:4 ratio), both PHA and PWM became mitogenic to B lymphocytes. Irradiated T lymphocytes alone did not respond to either PHA or PWM, indicating that the 3H-thymidine incorporation seen in the mixed-cell culture was due to the activation of unirradiated B lymphocytes. The B lymphocytes failed to respond to these phytomitogens in the presence of lower concentrations of irradiated T lymphocytes. The monocytes were found to be incapable of helping the B lymphocytes to respond to PHA or PWM.


The generation of memory cells. II. Generation of B memory cells with preformed antigen-antibody complexes. Mice were immunized with preformed complexes of dinitrophenylated haemocyanin (DNP-KLH) and anti-DNP or anti-KLH antibodies, and subsequently assayed for the generation of B memory (BM) cells. Complexes formed at equivalence or in slight antigen excess were far more effective than antigen alone in generating memory: as little as 100 ng DNP-KLH-anti-DNP produced substantial B cell priming. Anti-carrier and anti-hapten antibodies were equally effective. Complexes generated memory more rapidly than antigen alone, and the adjuvant effect was not simply due to aggregation of the antigen. Optimal priming by complexes required the integrity of the Fc portion of the antibody: F(ab')2 antibody fragments were less effective. The capacity of complexes to prime BM cells was abrogated by depriving mice of C3. C3 was also required for localization of complexes within splenic lymphoid follicles; complexes made with F(ab')2 localized in follicles, but less efficiently than those made with intact antibody. These results extend earlier findings (Klaus & Humphrey, 1977) and strongly suggest that the generation of BM cells involves the C3-dependent localization of antigen-antibody complexes within lymphoid follicles and strengthen the concept that germinal centres are the birthplace of BM cells.


Macrophage-lymphocyte association in in vitro mouse spleen cultures; the formation of B-cell colonies. Colonies of predominantly B lymphocytes have been grown in liquid culture, without agar or specific mitogen treatment. An essential component of the colonies is a macrophage-like cell. Colonies appear to result from continued growth of B lymphocytes following a primary macrophage-lymphocyte association which frequently occurs under a variety of culture conditions. Factors promoting colony growth are foetal calf serum and 2 mercapto-ethanol, the latter acting in part through reduction of a non-dialysable sulphur-containing component of serum.


Activation of B lymphocytes by mycoplasma mitogen(s). Various strains of the murine mycoplasma M. neurolyticum have been shown to induce extensive blast transformation of mouse lymphocytes, comparable in strength to mitogenicity exerted by these mycoplasma species on rat lymphocytes. The data summarized in this report demonstrate that this mitogenic effect is non-specific. Lymphoid cells from mycoplasma free, germ-free mice were activated to the same extent as those lymphocytes obtained from conventionally bred animals. Lymph node cell suspensions obtained from athymic nude mice were strongly activated by M. neurolyticum mitogen. Furthermore, mouse thymocytes and mouse T-cell enriched populations, were not stimulated by these mitogens. It was thus suggested that M. neurolyticum activates mouse B lymphocytes.


Active E rosette formation by human lymphoblasts. The percentage of active and total E rosettes for lymphocytes cultured for 72 h both with and without phytohaemagglutinin (PHA) was evaluated in 20 normal subjects. A clear increase in the formation of active E rosettes by PHA-stimulated lymphocytes was demonstrated with respect to unstimulated lymphocytes. Our data gives further proof that the active E rosettes constitute a specific subpopulation of lymphocytes which represent functionally active T cells in the blood.


T-lymphocyte colonies in the lymphoproliferative disorders. Human lymphocytes from peripheral blood, bone marrow spleen and lymph nodes were cultured. Continuous phytoheamagglutinin (PHA) stimulation was used, first during a 24 h liquid preincubation, then during a 5 day culture in methylcellulose. In normal donors a rapid colony formation took place, with a mean of 124+/-82 colonies per 1 times 10(5) preincubated lymphocytes. Cells from such colonies were studied by cytology, scanning electron microscopy and rosette formation techniques; arguments favour the hypothesis that these could be T lymphocytes. Neither granulocytes nor macrophages could be grown, and no lymphoid colony formation occurred without PHA stimulation. The same technique was applied to patients with various lymphoproliferative disorders. Significant colony suppression was observed in nearly every case of chronic lymphatic leukaemia; the number of colonies was reduced in some patients with acute lymphatic leukaemia, lymphosarcoma, dysglobulinaemia and Hodgkin's disease. This lymphoid culture method should be applied to a larger number of patients to determine whether it has a classification value and/or prognostic significance. When colonies were grown in pathological states, rosette formation was identical to that of normal donors; colony formation could be due to persisting normal lymphocytes.


Distinction between suppressors of the delayed-type hypersensitivity and the humoral response to sheep erythrocytes. Suppressors for both delayed-type hypersensitivity (DTH) and the humoral immune response could be simultaneously induced in the spleens of mice by immunization with a high dose of SRBC. Normal recipient mice of the spleen cells from donors immunized 5 days previously elicited depressed DTH or humoral response when immunized with SRBC. The suppressive activity was found to reside in T not B enriched fraction. Four hundred rad irradiation of the primed spleen cells resulted in complete loss of DTH suppressor activity, but only in some reduction of the suppressor activity for the humoral response. In contrast, hydrocortisone treatment of the donor mice caused no loss of DTH suppressor activity while approximately half of the suppressive activity for anti-SRBC PFC response was lost. Adult thymectomy prevented completely the induction of the DTH suppressor in contrast to little loss of the suppressor activity for the humoral response. DTH suppression was antigen-specific for the induction, but nonspecific for the expression. However the suppression of the humoral response was antigen-specific not only for the induction but also for the expression. In addition, DTH suppressor was capable of suppressing both the induction and expression of DTH while the humoral response was suppressed only in the induction stage by the suppressor.


Isolation of various canine leucocytes and their characterization by surface marker analysis. Various techniques were used to separate canine peripheral blood leucocytes into populations enriched in lymphocytes, polymorphonuclear leucocytes, phagocytic mononuclear cells (monocytes) and macrophages. Surface markers on each cell population were determined by rosette formation. Fc receptors for IgG and complement receptors (C3b and C3d) were present on PMN, monocytes, macrophages as well as on a sub-population of lymphocytes. Purification of the lymphocytes into T-and B-cell-enriched populations revealed that these receptors were present only on the B lymphocytes and not on the T lymphocytes. In addition, a third lymphocyte population, which did not possess surface immunoglobulin, and Fc receptor but not the complement receptor. None of the cell populations exhibited C4 complement receptors or Fc receptors for IgM. When different cell populations were tested for their ability to form rosettes directly with human type 'O' red blood cells it was found that most populations could rosette, suggesting that this technique could not be used as a specific marker for canine T lymphocytes.


Expression of T cell suppressor activity in the immune response of newborn mice to a T-independent synthetic polypeptide. One and two day old mice responded to the T-independent synthetic polypeptide poly (DTyr,DGlu)-poly(DPro)--poly(DLys) with high antibody production, whereas a dramatic decrease was observed in the immune response of 3 day old injected animals. The presence and expression of suppressor T cells in the 3 day old newborn mice spleens was demonstrated by transferring spleens of 3 day old mice into either thymectomized, irradiated recipients together with bone-marrow cells, or into 1--2 day old newborn recipients. In both cases a significantly lower antibody response was observed in recipients of the 3 day old newborn spleen cells as compared to recipients of the same number of adult spleen cells. No decrease in the antibody levels was detected following inoculation of 3 day old spleen cells after anti-theta-treatment, while enriched T cell populations obtained from nylon wool columns had the same suppressive effect as the whole spleen cell preparations. Thus, the cell expressing the suppressor activity in the newborn spleen is a T cell.


Studies on the host-parasite relationship in Schistosoma mansoni-infected mice: the immunological dependence of parasite egg excretion. CBA mice deprived of their T cells by means of thymectomy and anti-thymocyte serum and subsequently infected with Schistosoma mansoni were found to have substantially fewer parasite eggs in their faeces than similarly infected immunologically-intact control animals. The number of parasite eggs deposited in the tissues of T-cell deprived mice was by comparison only marginally lower than in control mice. Administration of serum obtained from normal mice with chronic S. mansoni infections partially restored the egg excretion rate in infected deprived mice, and also resulted in an increased number of eggs being deposited in the liver and intestine of these animals.


Effects of the B-cell activators lipid A and dextran sulphate on the antibody response to sheep red blood cells in piglets. The in vivo effects of Lipid A and dextran sulphate on the antibody response to the thymus-dependent antigen SRBC in piglets are reported. No alteration of the low primary antibody response was observed with any of the B-cell activators. However, both substances induced an increase in antibody titre after secondary challenge with the antigen, which was shown to be of the IgG class. An activation of immunological memory in the absence of primary antibody response is suggested.


Studies of hyperthymic mice. I. The influence of multiple thymus grafts on the size of the peripheral T cell pool and immunological performance. Following the implantation of syngeneic chromosomally marked thymuses under the kidney capsule of normal adult mice, donor T cells were found in the blood and other lymphoid organs. The mice were found to have increased numbers of Thy.1-positive cells in spleen and lymph nodes, increased peripheral blood lymphocyte levels and also augmented immune responses to sheep erythrocytes. The contribution made by an individual graft to the donor T cell pool was independent of the number of thymuses grafted as was the growth of the grafts themselves. The results reaffirm that there is no homeostatic feedback control of thymus growth in adult mice.


Surface membrane changes of T cells induced by syngeneic tumour cells. II. T-cell defects induced by small tumour cell inocula or tumour cell antigens. Injection of a large number of tumour cells, like other strong immunogenic challenges, is followed within 6 h by the uptake of cytophilic Ig (probably complexes) by a subpopulation of T cells. This phenomenon, known as the "6-hour T-cell response" is abrogated when small tumour cell inocula (10(2)), or small amounts of a preparation from tumour cells, which contains tumour antigens, are injected prior to the immunogenic challenge Abrogation of the "6-hour T-cell response" resulted in a decrease in specific anti-tumour cell immunity as tested in vitro by measuring growth inhibition (cytostasis). It has also resulted in loss of the amplifying function on antibody formation against sheep erythrocytes, normally detected in a T-B cell co-operative system when T cells are used 6 h after priming with sheep erythrocytes. It is postulated that this T-cell defect may represent a mechanism by which tumour cells, in the early stages of their growth, interfere with inductive stages of the immune response for a sufficient period of time to allow the tumour to grow beyond immune control.


Human tumor-lymphocyte interaction in vitro. III. T lymphocytes in autologous tumor stimulation (ATS). The T-cell nature of the responding cells in autologous tumor stimulation (ATS) was proven by (1) E-rosette formation of lymphocytes attached to tumor cells during the early period of co-cultivation; (2) blast-transformed E-rosetting cells detectable at the end of the test period (day 6) and (3) the reactivity of T-enriched lymphocyte subset and unresponsiveness of T-depleted fractions. Preparation of tumor suspensions devoid of lymphocytes made it possible to carry out cross-tests between patients. Blastogenesis with allogeneic tumors was rare. The restricted autologous nature of the reaction is a strong indication for genuine tumor-relatedness.


Heteroantiserum against acute lymphocytic leukemia raised to the lymphoblastoid cell line NALM-1. Antisera have been raised in rabbits to the lymphoblastoid cell line NALM 1 precoated with anti-lymphocyte serum (ALS). Following absorption with chronic lymphocytic leukemia cells (CLL) the antisera reacted mainly with acute lymphocytic leukemia (ALL) cells, and were very similar in specificity to antisera raised to ALL cells precoated with ALS. Leukemia cells from the following numbers of patients were positive for the anti-NALM 1 sera in a complement-dependent cytotoxicity test; 11/14 ALL, 3/15 acute myelocytic leukemia (AML), 1/5 chronic myelocytic leukemia (CML) and 0/8 CLL. Normal B and T peripheral blood lymphocytes were negative. The titer of the anti-NALM 1 sera against positive cells was 1:64 to 1:256 whereas the undiluted sera did not react with negative cells. Ten out of 11 of the positive ALL cells were of the non-B non-T type. However, cells from 1/4 T ALL patients and a cultured T ALL line 8402 were also positive. Six of 12 cultured lymphoblastoid cell lines were positive, all of which were of malignant origin. The molecular weight of the ALL antigen detected by anti-NALM-1 serum was determined by immunoprecipitation and sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) to be approximately 98,000 daltons.


Effect of 60Co gamma radiation on the valyl-tRNA synthetase isolated from chick embryo brain. The effect of 60Co gamma irradiation on the activity of valyl-tRNA synthetase isolated from chick embryo brain was studied. The enzyme activity exponentially decreased in the dose range 10--200 krad. The first step of the enzyme action, i.e. the amino acid activation, was found to be less sensitive to irradiation than the whole reaction, the formation of valyl-tRNA, 2-Mercapto ethanol and/or glycerol had a significant radioprotective effect. The lesion caused by radiation in the enzyme was also influenced by its concentration during exposure (dilution effect). According to gel-electrophoretic experiments, no chain rupture occurred in the enzyme molecule. Not even a change in Km was observed; however, the maximum velocity of the reaction was found to decrease with increasing radiation dose.


Chromosome condensation and radiation-induced G2 arrest studied by the induction of premature chromosome condensation following cell fusion. When mitotic and interphase cells are fused together, the chromosomes of the interphase cell sometimes condense prematurely. The phenomenon of premature chromosome condensation (PCC) was utilized in investigating the problem of whether the chromosomes of cells suffering a radiation-induced G2 delay are capable of condensation. Colcemide-arrested mitotic cells were fused with synchronized G2 cells, and with irradiated cells suffering a G2 delay. The frequency of PCC in mitotic X G2 binucleate cells was determined. This was compared to the PCC frequency in an unirradiated synchronized population rich in G2 cells after fusion with mitotic cells. Flash-labelling with 3HTdR and autoradiography allowed us to eliminate S-phase cells. The frequency of G2 PCCs was not significantly different for the irradiated G2-delayed or unirradiated cells. From these results we conclude that the chromosomes of cells suffering a G2 arrest are capable of condensation, although the involvement of the condensation process in radiation-induced G2 delay cannot be ruled out.


Radiation-induced damage to blood vessels: a study of dose--effect relationship with time after X-irradiation. Radiation damage to the right femoral artery was studied in mice 7--8 weeks old. Single X-ray exposures of 500 to 10 000 rad were used. The animals, except for the right upper legs, were shielded from exposure. The unexposed left femoral arteries served as 'controls'. Animals were killed at post-irradiation times of up to 120 days. Pathological changes in the irradiated arteries were scored on an arbitrary scale of 0 to 5 for individual tunica. The overall arterial damage was scored on a scale of 0 to 15. Radiation doses of 1000 to 10 000 rad produced quantitative histological changes in the three individual tunica and the degree of damage was dose-dependent. The tunica adventitia showed the maximum radiation injury, followed by tunica media and tunica intima in decreasing magnitude of injury. The combined effects on the artery followed closely the effects on media and adventitia.


Syntheses of DNA and chromosomal proteins in the rat thymus following whole-body X-irradiation. Syntheses of chromosomal proteins was studied in relation to DNA syntheses in the rat thymus following whole-body X-irradiation (600 rad). There was a considerable depression of the syntheses of DNA and histones during the first 4--48 hours after irradiation. The syntheses of histones H3 and H4 plus H2A were affected to a much greater degree than those of histones H1 and H2B, suggesting a tight coupling between the syntheses of DNA and histones H3 and H4 plus H2A. A part of the lysine-rich histones H1 and H2B, however, seems to be synthesized, even in the absence of DNA synthesis. Biosynthesis of non-histone proteins was also depressed in the thymus after irradiation. The degree of inhibition, however, was very low, except for the syntheses of a few non-histone protein components which were depressed to a considerable extent. This implies that the synthesis of the majority of non-histone proteins is independent of DNA synthesis.


Medically and surgically treated coronary patients in cardiovascular rehabilitation: a comparative study. Mood and physical performance were assessed in medically and surgically treated patients with coronary artery disease referred to a cardiovascular rehabilitation program early in convalescence. Surgically treated patients reported less mood disturbance prior to training and demonstrated greater gains in exercise tolerance. Medically managed patients had more mood disturbance prior to training and greater mood improvement in the six week rehabilitation program. Improvements in mood were independent of improvement in exercise tolerance. Group differences in mood state could not be attributed to the use of defensive denial, time from discharge, severity of coronary disease, or age of patient. The psychological and physical impact of bypass surgery contributes to the mood and physical performance of postoperative patients.


[Ulcus-molle epidemic in Turkey].

[Ulcus-molle epidemic in Turkey]. 240 patients with soft chancre who have been observed within a year dating from March 1976 at the University Dermatological Clinic of Istanbul are presented. The clinical findings encountered in our cases are presented and the therapeutic results are discussed.


[Protein concentrations in the ejaculate of patients with chronic prostatitis]. In 158 patients suffering from chronic prostatitis and 23 healthy men seminal fluid concentrations of IgG, IgA, C3c, coeruloplasmin, alpha-1-antitrypsin and alpha-1-glycoprotein were determined by immunodiffusion. Concentration of IgG is closely correlated to leucocyte counts, that of IgG, C3c, and coeruloplasmin to erythrocyte counts. C3c, coeruloplasmin, alapha-1-glycoprotein and alpha-1-antitrypsin are further correlated to sperm counts. IgG and C3c concentrations are related to pathogenetic factors. If pathogenic bacteria are found, they are significantly higher than in patients with mycoplasmas, without bacteria or normal men. The same relation is found when observing the leucocyte count. IgG is significantly lower when mycoplasmas are found.


Ultrastructural localization of alkaline phosphatase in the hypertrophic chondrocyte of the frog. The ultrastructural localization of alkaline phosphatase was studied in the hypertrophic chondrocyte of the frog (Rana temporaria) by incubating sections of glutaraldehyde fixed tissue in a medium containing sodium beta glycerophosphate and calcium chloride. Control specimens were incubated in substrate free medium. Alkaline phosphatase (orthophosphoric monoester phosphohydrolase) is a high molecular weight glycoprotein that hydrolyses phosphorylated metabolites much as acid phosphatase does except that its action is optimal at an alkaline pH. The results of this investigation showed that alkaline phosphatase activity was present within the cytoplasm and around the plasma membrane of frog hypertrophic chondrocytes. Although only a small proportion of frog hypertrophic chondrocytes demonstrated enzyme activity, there was evidence that this was concentrated within Golgi lamellae and vesicles leaving other organelles unreactive. The finding of alkaline phosphatase activity within Golgi lamellae of hypertrophic chondrocytes is regarded as unusual although postitive reactions within chondrocyte lysosomes have previously been reported (Doty and Schofield, 1976).


[Congenital cysts of the oval window (author's transl)]. A 3 year old child with recurrent pneumococcal meningitis and a congenital cyst of the oval window is presented. An aqueous middle ear effusion was found and scintigraphy demonstrated a middle ear CSF leak. Surgical exploration revealed a cyst originating from the stapes footplate. The origin of this malformation is discussed and its possible presence in recurrent mengitis is stressed.


Apparent suicide inactivation of human lymphoblast S-adenosylhomocysteine hydrolase by 2'-deoxyadenosine and adenine arabinoside. A basis for direct toxic effects of analogs of adenosine. Adenosylhomocysteine hydrolase from human lymphoblasts binds 2'-deoxy[3H]adenosine tightly. Binding is associated with time-dependent, saturable, irreversible inactivation of catalytic activity which occurs with first order kinetics, suggesting "suicide" inactivation. Adenine arabinoside produces similar inactivation but is more potent. These results suggest a basis for a heretofore unrecognized mechanism of action for these and other analogs of adenosine, in which toxicity results from actions of the nucleosides themselves, rather than from nucleotides to which they may be converted.


The sidewinder catheter for extra cranial angiography: technical, diagnostic and therapeutic applications. Through using the sidewinder catheter system for visceral angiography in over 200 patients we have improved our success rate. The method is simple and safe, and we recommend it.


Glomerular permeability in the bullfrog Rana catesbeiana. Filtration studies suggest similar size pores in the glomerular filters of mammals and amphibians. However, the glomerular wall in the bullfrog exhibits several structural features not found in mammals. The subendothelial space of the basement membrane is often greatly enlarged and infiltrated by cellular elements. The lamina densa of the basement membrane shows extensive variation in thickness and packing of its filaments. On the other hand, the epithelial slits in the bullfrog are closed by a slit diaphragm which appears similar in size and structure to the slit diaphragm in mammals. Horse spleen ferritin, a protein with a hydrodynamic radius of 61 A, was used as an ultrastructural tracer to determine whether the highly variable structure of the basement membrane renders this layer more permeable than its mammalian counterpart. Within 10 min after intravenous injection, ferritin was found throughout the basement membrane and often in clusters within the subepithelial layer adjacent to the slit diaphragm. Virtually no ferritin was found within the urinary space, podocytes, or cells of the proximal tubule. Ferritin distribution was the same in both superficial glomeruli and more deeply lying glomeruli regardless of the method of fixation. These results indicate that in the bullfrog the slit diaphragm is a principal filtration barrier to ferritin and thus to smaller plasma proteins.


Two distinct mechanisms for redistribution of lymphocyte surface macromolecules. I. Relationship to cytoplasmic myosin. A detailed kinetic analysis of the distribution of cytoplasmic myosin during the capping of various lymphocytic surface molecules revealed two distinct capping mechanisms. (a) Some cell surface molecules, including immunoglobulin, Fc receptor, and thymus leukemia antigen, all cap spontaneously in a small fraction of lymphocytes during locomotion. Cytoplasmic myosin becomes concentrated in the cytoplasm underlying these spontaneous caps. Exposure to specific antibodies causes all three of these surface molecules to cap rapidly with a concomitant redistribution of cytoplasmic myosin to the area of the cap. These antibodies also stimulate cell locomotion. (b) Other lymphocyte surface molecules, including H2 and Thy.1, do not cap spontaneously. Moreover, exposure to antibodies to these molecules causes them to cap slowly without a redistribution of cytoplasmic myosin or stimulation of cell locomotion. Exposure to concanavalin A gives a response intermediate between these two extremes. We believe that the first type of capping is active and may involve a direct link between the surface molecules and the cytoplasmic contractile apparatus. The second type of capping appears to result simply from aggregation of cross-linked molecules in the plane of the membrane.


A collision analysis of lymphoid cell aggregation. We have obtained data on the frequency of aggregates of different size classes and on the maximum aggregate diameter of lymphoid cells subjected to aggregation in the laminar shear field of a Couette viscometer. Maximum aggregate diameter reached a plateau level during aggregation. This plateau is considered to be the result of a balance between the hydrodynamic shear forces tending to resist the formation of aggregates, and the adhesive forces of the aggregated cells tending to resist dissociation. Stepwise-increases in the shear rate produced aggregates of progressively smaller maximum diameter until the limiting (i.e. control equivalent) diameter was reached. Equations governing these stepwise changes in aggregate diameter were obtained by regression analysis, and an estimate of the force of dissociation (FD) was made from derived values of the critical shear rate. Thymocytes (FD=2.54 X 10(-6) N m-2/cell) were found to be more adhesive than lymphocytes (FD=2.05 X 10(-6) N m-2/cell), in agreement with current concepts. The observed data on aggregate frequency were seen to be of poor fit with a model of aggregation derived by collision analysis of the aggregation process. This led us to consider the possibility that all cells may not share the same probability of forming an adhesion. We thus derived further models of aggregation in which some fraction of the total cells was considered to have enhanced possibilities of a collision producing an adhesion. Of the models we considered, a 15% preferred fraction offered best agreement with the experimental observations. We therefore conclude that the populations of cells studied in this report are not 'homogenous' in that some cells are more adhesive than others. Alterations in the percentage of the preferred fraction of cells will lead to different aggregate-frequency indices. Such changes might be expected to occur during the initial stages of carcinoma development.


Oxygen uptake during the cell cycle of the fission yeast Schizosaccharomyces pombe. Oxygen uptake was measured in synchronous cultures of the fission yeast Schizosaccharomyces pombe. The rate of oxygen uptake was found to increase in a step-wise manner at the beginning of the cycle and again in the middle of the cycle. The increases in rate were such that overall, oxygen uptake doubled in rate once per cell cycle. Addition of inhibitors of DNA synthesis or nuclear division to a synchronous culture did not affect the uptake of oxygen. In an induced synchronous culture, in which DNA synthesis, cell division, and nuclear division, but not 'growth' were synchronized, oxygen uptake increased continuously in rate and did not show the step-wise rises which were shown in the selection-synchronized culture. These results were compared with previous measurements of oxygen uptake in yeast and an explanation is suggested for the many different patterns which have been reported.


Nerve fibers of the eighth nerve and their distribution to the sensory nerves of the inner ear in the bullfrog. The number of both myelinated and unmyelinated nerve fibers has been determined for the eighth nerve and each of its sensory branches in the bullfrog. No significant differences were found in the size distribution of the myelinated nerve fibers among the three ampullary nerves or between the saccular and lagenar nerves. Comparison of these two functional groups also showed no significant differences in the myelinated nerve fiber size distribution between the angular acceleration and otolithic receptors, collectively the equilibrium receptors. The wide variability, that was found not only among the animals but also between the right and left sides, most likely resulted in the absence of differences. Highly significant differences were noted when the myelinated nerve fiber diameter distributions to the acoustic organs were compared to those of the equilibrium organs. The most notable difference was the absence of any myelinated nerve fibers whose diameters were greater than 8 micrometer.


A histochemical study on the innervation of cerebral blood vessels in the bullfrog. Specific histochemical techniques for the demonstration of catecholamine and acetylcholinesterase have been used to study the distribution of adrenergic and cholinergic nerves on the cerebral blood vessels of bullfrog, Rana catesbeiana. The adrenergic nerve meshworks on the cerebral arteries of bullfrog were less dense, had a more elongate appearance along the arterial axis as compared with those of mammals and were rather similar to those of snakes. The nerve plexuses on the cerebral carotid artery and its main branches were somewhat denser than those on the basilar artery. The most characteristic feature of innervation in the bullfrog cerebral vessels was that no acetylcholinesterase-positive fibres were observed on the extraparenchymal arteries, whereas, in all higher vertebrates investigated so far, the cerebral arteries have been found to be dually innervated although differences in the density of innervation of the two nerves may exist. This suggests that the peripheral adrenergic innervation on the cerebral blood vessels appeared earlier than the cholinergic one in the evolution of vertebrates. On the other hand, both adrenergic and acetylcholinesterase-positive fibres were observed in close contact with parenchymal arterioles and capillaries suggesting the possible existence of a dual central innervation. This feature, however, was by no means common. Thus, the central neurons have a significant influence on the cerebral circulation in the bullfrog is somewhat equivocal. Most of the pial and the parenchymal small vessels and the parenchymal capillaries exhibited a heavy acetylcholinesterase activity on the vascular walls. Although the significance of the enzyme is obscure as yet, this has to be considered in relation to the regulatory mechanism of the cerebral circulation.


Visualization of intracellular concanavalin A binding sites in retinal photoreceptors. Localization of carbohydrate components in retinal photoreceptor cells and membranes was studied. Frog and rat retinas were fixed with glutaraldehyde and embedded in glycol methacrylate or in a mixture of glycol methacrylate, glutaraldehyde and urea. Thin sections were incubated with ferritin-labeled concanavalin A (F-Con A) and stained with osmium vapors. Intensive binding was observed in both rod and cone outer segments. In the rod inner segment, differential binding of F-Con A was demonstrated. While numerous ferritin granules were observed in the myoid zone, only a few were seen in the ellipsoid zone, except for a local accumulation along the plasma membrane. In the rod outer segment, Con A binding sites were closely associated with the disk membranes. Ferritin granules were observed on both sides of the membranes. The relationship between the localization of Con A binding sites and the orientation of visual pigment molecules within the rod outer segments disk membranes was discussed.


Proliferative response to anti-IgM antibodies of various B lymphocyte subpopulations isolated by cell sorting. We have utilized several methods to isolate populations of murine splenic B lymphocytes bearing different classes of surface Ig molecules. Antibody and complement-mediated cytotoxicity was used to remove IgD-bearing B cells, and fluorescein-conjugated anti-IgM and polyspecific anti-Ig-F(ab')2 antibody fragments were used with a fluorescence-activated cell sorter to isolate cell bearing IgM or all Ig classes. First, the technique of isolating cells in the sorter was evaluated. It was found that among cells sorted as negative for Ig there was always a certain percentage where IgM reappeared on the membrane after overnight culture. Second, evidence is presented that surface IgM is present on the controls the proliferation of the majority of B lymphocytes in an adult mouse spleen. This includes cells bearing predominantly IgM and cells bearing predominantly IgD. Finally, a theory of B cell stimulation is presented, based on distinctive roles for surface IgM and IgD molecules, as well as a balance between their signals to an individual cell.


Interaction of cytotoxic T lymphocytes with target cells. I. Specific inhibition by detergent-solubilized, partially purified mouse histocompatibility antigens. H-2 antigens from EL4 (H-2b) and RLmale 1 (H-2d) leukemias were solubilized with deoxycholate and partially purified based on their adherence to a Lens culinaris hemagglutinin column. In double reciprocal experiments we have shown that these preparations specifically inhibit conjugate formation between target cells and alloimmune peritoneal lymphocytes, a preparation rich in cytotoxic T lymphocytes.


The effects of anti-IgD on serum immunoglobulins, antibody production, and immunoglobulin-bearing cells in adult rats. Rabbit anti-rat IgC (delta-specific) injected subcutaneously into adult rats resulted in hypergammaglobulinemia, increased serum immunoglobulins, increased antibody production, and alterations in the percentages of PBL expressing IgD and IgM. Serum IgM, IgG2a and IgG2c concentrations were increased 100%, 125%, and 200%, respectively, over controls, whereas IgA levels were not altered. Increases in the serum immunoglobulins were predominately due to an increase in antibodies to the rabbit serum components. A depletion of IgD-IgM bearing lymphocytes in the peripheral blood was observed after 24 hr of injection. Percentages of IgC-IgM bearing lymphocytes returned to normal by day 4 and were significantly increased (approximately 2-fold) by day 8. The in vivo effects of anti-delta in rats were similar to those reported in the monkey. Possible modes of action of the anti-delta and the significance of these results in relation to the biologic function of membrane IgD are discussed.


5'-Nucleotidase activity in subpopulations of rat lymphocytes. Rat lymphocyte populations were assessed for 5'-nucleotidase activity by both biochemical and histochemical methods. Enzyme-specific activity was enriched in lymphocyte plasma membrane fractions and was higher in lymph node and spleen lymphocytes than in thymocytes. Histochemical reactions on sections of spleen and lymph node revealed strong activity in the thymus-dependent regions, periarteriolar lymphoid sheath in spleen, and paracortex in lymph node; whereas the thymus-independent regions, follicles, and germinal centers were negative. In cellular depletion experiments, with three different methods to detect 5'-nucleotidase, it was observed that the depletion of T cells, but not B cells, was accompanied by a loss of enzyme activity and a decrease in the percentage of nucleotidase-positive cells. The results suggest that, among members of the lymphocyte series, high 5'-nucleotidase activity is selectively associated with the plasma membranes of peripheral T cells in the rat.


Modulation of immune response by bacterial lipopolysaccharide (LPS): cellular basis of stimulatory and inhibitory effects of LPS on the in vitro IgM antibody response to a T-dependent antigen. The role of thymus-derived lymphocytes (T cells) in LPS modulation of T cell-dependent antibody responses has been investigated. We have assessed the effect of LPS on the primary anti-TNP response to TNP-SRBC of cultures of whole spleen cells or T cell-depleted spleen cells that were supplemented with various subpopulations of carrier-primed (SRBC) spleen cells. The TNP-PFC response was enhanced in the presence of irradiated SRBC-primed spleen cells by addition of 0.16 to 20 microgram/ml LPS, but inhibition was observed when irradiation of primed cells was omitted. Enhancement but no inhibition occurred when added primed cells were first passed through a nylon wool column. LPS-mediated enhancement was dependent on a T cell in the primed population. These results suggest that LPS modulation of antibody synthesis is dependent on two populations of antigen-specific cells that have opposing effects on B cell responses to a T-dependent antigen: a helper cell that is irradiation resistant, nonadherent to nylon wool, and sensitive to anti-T cell serum, and a suppressor cell that is irradiation sensitive and adherent to nylon wool.


Development of IgE-forming cells in vitro from rat mesenteric lymph node cells. Mesenteric lymph node cells from normal rats and rats infected with Nippostrongylus brasiliensis (Nb) were cultured with pokeweed mitogen (PWM) or Nb antigen, and the development of IgM-, IgG2a-, or IgE-containing cells was assessed by immunofluorescence. Normal lymph node cells stimulated with PWM developed into both IgM- and IgE-containing cells, whereas similar stimulation of cells from Nb-infected rats resulted in the development of IgM-, IgG2a-, and IgE-containing cells. The in vitro plasma cell response to PWM was dependent on the presence of T lymphocytes. Lymph node cells from Nb-infected rats responsed to Nb antigen and developed into plasma cells of IgM, IgG, and IgE classes. The response was antigen specific and required antigen-primed T cells. Depletion of IgE-bearing cells or IgM-bearing cells before stimulation with either PWM or Nb antigen diminished the level of IgE forming cell development, suggesting that IgE-IgM double bearing cells are precursors of IgE-forming cells. The distribution of the three isotypes among the If-forming cells that developed in response to PWM was influenced by the source of both B and T cells. When B cells from Nb-infected rats were employed as a source of precursors, T cells from infected animals were more effective than normal T cells for the development of IgE-forming cells, whereas the latter cells were more effective for the development of IgG2a-forming cells than T cells from infected animals.


Distribution of identity of Thy-1-bearing cells during ontogeny in rat hemopoietic and lymphoid tissues. The distribution and identity of Thy-1+ cells during ontogeny in the rat were examined by immunofluorescent and histologic techniques, and the relative amount of Thy-1 antigen on different cell types was assessed by quantitative absorption. Thy-1+ cells were subsequently studied by dual immunofluorescence for the simultaneous expression of either T or B lymphocyte markers. The results indicate that the Thy-1 antigen is present on hemopoietic precursors and on at least some of their lymphoid (both T and B) and nonlymphoid descendants. In addition, six serologically distinct subpopulations of lymphoid cells were identified, and their position in T and B cell maturational pathways is discussed.


Recognition by cytolytic T and K cells: identification in both systems of a divalent cation-independent, cytochalasin A-sensitive step. Most studies of recognition events in cell-mediated cytolysis have analyzed the conditions that are required to establish mechanically stable binding between effector cells and target cells. This approach has shown that cytolytic T cells require a temperature above 20 degrees C and free divalent cations to bind specifically to target cells. K cells on the other hand were shown to bind to IgG on target cells in the absence of divalent cations at 4 degrees C. In this paper we show that both K cell- and T cell-mediated cytolysis are inhibited by cytochalasin A, if this drug is added to cultures before the effector and target cells have interacted. However, the drug does not block the calcium-dependent lethal hit stage of cytolysis in either system. We then show that in both T cell- and K cell-mediated cytolysis, the cytochalasin A-inhibitable stage can proceed in the absence of divalent cations. We conclude from these data that: 1) both K cell- and T cell-mediated cytolysis have a divalent cation-independent stage, which is inhibitable by cytochalasin A; 2) firm mechanical binding of T effectors to target cells requires divalent cations. It is uncertain if the latter is necessary for cytolysis of whether K cells also have to go through a divalent cation phase before the lethal hit stage. The implications of these findings are discussed, especially as to the possibility that strong binding may not be necessary for T cell-mediated cytolysis.


The detection of alloantibodies to subpopulations of human lymphocytes: an adaptation of the indirect anti-immunoglobulin rosetting reaction (IARR). An indirect anti-immunoglobulin rosetting reaction (IARR) can be successfully used to detect alloantibodies to human lymphocytes. In this paper we describe an adaptation of the IARR which allows detection of alloantibodies to human lymphocyte subpopulations. Fluorescein labelled sheep red cells, as a T cell marker, are incorporated into the rosetting reactions and by looking for rosettes with two indicator cell types, sheep and ox, one can determine if the alloantibodies are reacting with T cells or non-T cells. This type of assay is more sensitive than a standard cytotoxic test and can detect non-cytotoxic antibodies. The results show that this rosetting assay with two types of indicator cell can be useful in the study of pregnancy anisera, particularly those that are thought to contain reactivity directed solely against non-T cells. These antisera probably recognise the human DR antigens which are thought to be equivalent to the rodent Ia antigens.


Spontaneous murine lupus-like syndromes. Clinical and immunopathological manifestations in several strains. MRL/1 and BXSB male mice have a systemic lupus erythematosus (SLE)-like disease similar to but more acute than that occurring in NZB X W mice. The common elements of lymphoid hyperplasia, B-cell hyperactivity, autoantibodies, circulating immune complex (IC), complement consumption, IC glomerulonephritis with gp70 deposition, and thymic atrophy were found in all three kinds of SLE mice. On the basis of these common elements, SLE seen in these mice can be considered a single disease in the same sense that human SLE is one disease. The differences in the SLE expressed in the different mice are no greater than those found in an unselected series of humans with SLE. However, the significant quantitative and qualitative variations in abnormal immunologic expression suggest that different constellations of factors, genetic and/or pathophysiologic, may operate in the three murine strains and that each constellation is capable of leading, via its particular abnormal immunologic consequences, to the activation of common immunopathologic effector mechanisms that cause quite similar SLE-like syndromes. From an experimental point of view, the availability of several inbred murine strains of commonplace histocompatibility types that express an SLE-like syndrome makes possible innumerable manipulations which should help to elucidate the nature and cause(s) of this disorder.


Anti-idiotype induced regulation of helper cell function for the response to phosphorylcholine in adult BALB/c mice. An adoptive secondary antibody response to phosphorylcholine (PC) can be generated by the transfer of keyhole limpet hemocyanin (KLH)-primed T cells, PC-bovine gamma globulin-primed B cells, and PC-KLH into irradiated syngeneic BALB/c mice. If the KLH-primed T-cell donors were pretreated with anti-idiotype antibodies directed against the BALB/c PC-binding myeloma TEPC 15, their T cells were unable to collaborate effectively with PC-primed B cells; moreover, they could suppress the helper activity of T cells from normal mice for the PC-KLH response. The Ly phenotype of these T cells was found to be Ly 1-, 2+. The specificity of the suppressor T-cell population induced by anti-T15 treatment appears to be both for idiotype (hapten) and carrier, since the suppressor T cells fail to interfere with the antibody response to PC on a heterologous carrier, nor do they suppress the response to trinitrophenol-KLH.


Evidence that cytotoxic T cells are part of the host's response to influenza pneumonia. Cytotoxic T cells were detected in the cervical lymph nodes, lungs, spleen, and peripheral blood of mice with influenza. Lymphocytes decreased in the peripheral circulation and increased in the lung during the period of acute inflammation and pneumonia. Peak cytotoxic T-cell activity was present at the time of marked pulmonary infiltration, and it decreased with resolution of the pneumonia. The cytotoxic T cells in the lung were shown to be H-2 restricted and specific for the hemagglutinin of the infecting virus. The results indicate that hemagglutinin specific cytotoxic T cells are (a) induced during influenza infection; (b) they circulate in the blood; (c) they are present in greatest number; and (d) they have their peak cytotoxic effect when pneumonia is most marked. We interpret the results to indicate that specific cytotoxic T cells in the infected target organ are part of the immunological and pathological response to virus infection.


Demonstration of Thy-1 antigen on pluripotent hemopoietic stem cells in the rat. Three approaches were used to demonstrate the presence of Thy-1 antigen on the surface of pluripotent hemopoietic stem cells in the rat. In the first, stem cells from fetal liver, neonatal spleen, and adult bone marrow were prevented from forming hemopoietic colonies in the spleens of irradiated recipients spleen (colony-forming unit assay) by incubation with antibodies to Thy-1 antigen. Highly specific rabbit heteroantiserum to purified rat brain Thy-1 antigen and mouse alloantisera to Thy-1.1-positive thymocytes were equally effective. This inhibition was neutralized by purified Thy-1 antigen. In a second series of experiments, Thy-1-positive and Thy-1-negative populations of nucleated bone marrow cells were separated by the FACS. All of the hemopoietic stem cell activity was recovered in the Thy-1-positive population. The stem cells were among the most strongly positive for Thy-1 antigen, being in the upper 25th percentile for relative fluorescence intensity. The relationships of Thy-1 antigen to the rat bone marrow lymphocyte antigen (BMLA) was shown in a third series of experiments. Rabbit anti-BMLA serum, which is raised against a null population of lymphocyte-like bone marrow cells, has been shown to have anti-stem cell activity. Here we demonstrate by double immunofluorescence, cocapping, and differential absorption studies that Thy-1 and BMLA are parts of the same molecule.


Ia-bearing T lymphocytes in man. Their identification and role in the generation of allogeneic helper activity. The presence of Ia-like antigens was demonstrated on a small population (2-6%) of highly purified human circulating T lymphocytes by immunofluorescence with a rabbit anti-Ia serum raised against the isolated bimolecular Ia structure. The Ia+ T lymphocytes have no surface or intracellular immunoglobulins. The expansion of this Ia+ T-cell population was encountered in certain patients. Ia antigens were also found on T blasts grown in long-term cultures with conditioned medium generated by phytohemagglutinin-stimulated lymphocytes. In addition, leukemia blasts which stained for Ia antigens and formed E rosettes were identified in the peripheral blood of two leukemic patients. This evidence further supports the existence of Ia-bearing T cells in man. The Ia+ T-lymphocyte population was shown to contain cells responsible for the generation of allogeneic helper activity. Elimination of Ia+ lymphocytes from a purified T-cell population by the anti-Ia antiserum and complement abolished its ability to help an allogeneic B-cell preparation to generate plaque-forming cells against sheep erythrocytes in vitro in the presence of the antigen.


Secondary induction of cytotoxic T lymphocytes with solubilized syngeneic tumor cell plasma membranes. Secondary induction of in vitro cytotoxic T lymphocytes in a syngeneic system has been achieved with plasma membrane, both in the particulate and solubilized forms. Both the induction and the lytic phases were shown to be immunologically specific. The effector cells generated were completely susceptible to treatment with anti-theta antibody and complement, suggesting that they are T lymphocytes.


HLA restriction of human cytotoxic T lymphocytes specific for influenza virus. Poor recognition of virus associated with HLA A2. Cytotoxic T lymphocytes (CTL), specific for influenza A/X31 virus, were generated from human peripheral blood lymphocytes. These CTL lysed target cells that were infected with the same virus and that shared HLA A or B locus antigens. Minimal lysis was observed when HLA-D antigens were shared. Not all HLA A and B antigens were equally effective. Efficient lysis of target cells was seen when HLA A1, A3, B7, B8, B27 and BW21 were shared with the CTL, but when HLA A2 was the only shared antigen lysis was usually minimal. This deficiency in CTL function associated with HLA A2 was not absolute. It is suggested that the function of this antigen might be influenced by other surface molecules on the cell and in particular the other HLA products.


Monoclonal origin of B lymphocyte colony-forming cells in spleen colonies formed by multipotential hemopoietic stem cells. Spleen colonies produced by transplanting lethally irradiated mice with either 12 day fetal liver or adult bone marrow cells were found to contain B- lymphocyte colony-forming cells (BL-CFC) . The proportion of BL-CFC positive spleen colonies did not increase substantially between 8 and 14 days after transplantation, the range being 18-45 percent. However, the absolute number of BL-CFC per spleen colony varied considerably (between 1 and 10,318), although the majority of colonies contained less than 200 BL-CFC. Irrespective of the time after transplantation, smaller spleen colonies were found to have a higher frequency of BL-CFC than larger spleen colonies. To determine the possible clonal origin of BL-CFC from spleen colony- forming unit (CFU-S), CBA mice were injected with equal numbers of CBA and CBA T(6)/T(6) fetal liver or adult bone marrow cells. Analysis of 7-15-day spleen colonies demonstrated that 90 percent were either exclusively T(6) positive or T(6) negative and approximately equal numbers ofboth colony types were observed. B-lymphocyte colonies were grown and successfully karyotyped from 19 spleen colonies. When compared with the original spleen colony karyotype the B-lymphocyte colony cells karyotype was identical in all 19 cases. In 3 of the 19 colonies analyzed a mixture of T(6) positive and T(6) negative karyotypes was present and identical proportions of the karyotypes were present in the pooled B-lymphocyte colony cells and spleen colony cells. The data indicate that the B-lymphocyte colony-forming cells detected in spleen colonies are genuine members of the hemopoietic clone derived from the initiating hemopoietic stem cell (CFU-S).


T-T-cell interactions during the vitro cytotoxic allograft responses. I. Soluble products from activated Lyl+ T cells trigger autonomously antigen-primed Ly23+ T cells to cell proliferation and cytolytic activity. Secondary murine cytotoxic T lymphocyte responses from alloantigen-primed T cells can be induced in vitro by apparently unrelated regimens, such as addition of either concanavalin A (Con A), conditioned medium from Con A stimulated lymphocyte cultures, conditioned medium from secondary mixed lymphocyte cultures (MLC), or stimulator cells sharing only the I-region with the stimulating cells used for primary sensitization. We now report that upon polyclonal (Con A), or antigen-specific (MLC) stimulation, Lyl+ T cells release a factor, which in turn triggers alloantigen primed Ly23+ T cells to proliferation and cytolytic activity. The secondary cytotoxic T lymphocyte inducing factor (SCIF) is produced within 24 h. For its production, an intact protein metabolism, not DNA metabolism, is required. Once induced, the functional activity of SCIF is nonspecific and not H-2 restricted. SCIF allows exponential growth and long-term propagation of cytolytic Ly23+ T cells with specificity to alloantigens used for primary sensitization. SCIF induced activation of alloantigen primed Ly23+ T cells does not require the presence of alloantigens. The results therefore reveal a process by which Lyl+ T-cell-derived nonspecific factor(s) induce autonomously Ly23+ T-cell-mediated, antigen-specific, cytotoxic T lymphocyte responses.


Immune suppression in vivo with antigen-modified syngeneic cells. I. T-cell-mediated suppression to the terpolymer poly-(Glu, Lys, Phe)n. The palmitoyl derivative of the linear polypeptide of poly-(L-Glu-L-Lys-L-Phe)n (GLphi) can be coupled to spleen cells directly. The intravenous administration of 2 X 10(5)--3 X 10(7) GLphi-coupled syngeneic spleen cells induces GL-phi-specific suppressor T cells in C57BL/6 nonresponder mice. The suppression is antigen specific and can be detected by the inhibition of the primary GLphi plaque-forming cell response to challenge with GLphi-fowl gamma globulin. The number of inducer cells required for suppression carry less than 0.1 microgram of antigen. Spleen cells from tolerized mice can transfer suppression to normal syngeneic recipients. The suppression is cyclophosphamide sensitive and the suppressor cells bear the Thy 1.2 marker. This method of inducing antigen-specific suppressor cells may be generally applicable to other antigen systems.


The immunological basis of endotoxin-induced tumor regression. Requirement for T-cell-mediated immunity. It was shown that although intravenous administration of bacterial endotoxin caused extensive hemorrhagic necrosis of four different syngeneic murine tumors, only two of these tumors subsequently underwent complete regression: the two that were shown to be immunogeneic as classically defined. An immunologic basis for endotoxin-induced regression was further indicated by the additional findings that regression, but not hemorrhagic necrosis, of the two immunogenic tumors failed to occur in mice that were immunodepressed by whole-body gamma-irradiation, or that were made T-cell deficient by thymectomy and irradiation. That endotoxin-induced regression is T-cell mediated was suggested by the findings that tumor regression was followed by a state of long-lived immunity to a tumor cell challenge implant, and with the possession by the host of T cells that were capable of passively transferring this state of immunity to normal recipients. It is concluded that although parenteral injection of endotoxin causes hemorrhagic necrosis of most solid murine tumors, it is only those tumors that are immunogenic enough to evoke the generation of T-cell-mediated immunity which subsequently go on to completely regress.


The immunological basis of endotoxin-induced tumor regression. Requirement for a pre-existing state of concomitant anti-tumor immunity. It was shown that of four syngeneic, murine tumors investigated, only those that evoked the generation of a state of concomitant anti-tumor immunity were susceptible to endotoxin-induced regression. Moreover, the temporal relationship between the generation of concomitant immunity and the onset of susceptibility to endotoxin-induced regression points to the likely possibility that tumor regression depends on the preceding acquisition of the specifically-sensitized, effector T cells that express concomitant immunity. It is suggested that endotoxin-induced hemorrhagic necrosis which invariably precedes tumor regression serves to create conditions inside the tumor that are conducive to the entry and the functioning of effector T cells. It is also suggested that tumor necrosis factor causes hemorrhagic necrosis rather than tumor regression.


Induction of in vitro differentiation and immunoglobulin synthesis of human leukemic B lymphocytes. Successful induction of in vitro differentiation and immunoglobulin synthesis of the leukemic lymphocytes was carried out in two cases of chronic lymphocytic leukemia. Few plasma cells and little specific Ig secretion were detected in the cultures of isolated leukemic B cells in either the presence or the absence of autologous T cells. Up to 30% of the leukemic B cells matured to plasma cells, and a 32-fold increase in specific Ig synthesis was observed when T cells from normal individuals were added to the cultures of these leukemic B cells. In one of the two cases, autologous T cells were able to induce greater than 50% of the leukemic B cells to differentiate further to plasma cells in the presence of pokeweed mitogen. This markedly accelerated in vitro differentiation was only achieved with leukemic cells from cases in which there was evidence of slight differentiation in vivo. No evidence could be obtained for excessive suppressor T cells in these patients. However, a T-cell defect in the generation of allogeneic effect helper factors was identified. This defect may be responsible for the reduced rate of leukemic maturation in vivo.


In vitro generation of antigen-specific helper T cells that collaborate with cytotoxic T-cell precursors. Antigen-specific helper T cells are required in the generation of cytotoxic T cells from thymocyte precursors. We have demonstrated that these alloantigen-specific helper cells can be generated in vitro and that both the quantity and quality of the helpers appear to be superior to the help obtained from unprimed spleen cells. Optimal helper cell activity is produced at day two of culture when CBA splenic helper precursors are stimulated by irradiated allogeneic spleen cells. Helper cell precursors are antigen-specific cells which cannot be instructed to express forbidden receptor specificities and bear theta antigen on their surface. The helper effectors are radioresistant, theta-bearing, and antigen-specific cells.


B-cell precursors specific to sheep erythrocytes. Estimation of frequency in a specific helper assay. A sensitive, specific, and reproducible in vitro helper assay is described which is suited to limiting dilution analysis of murine B cells. 1 in about 3,000 syngeneic splenic B cells can be induced to form plaque-forming cells (PFC) to sheep erythrocytes in this system. The induction of PFC is absolutely dependent on antigen  and specific helper T cells.


Nonspecific signals for B-cell localization and activation. Virgin, inactive mammary gland autografted to the anterior chamber of the rabbit eye remains free of lymphoid cells. Activation of the ectopic gland by systemic injection of chorionic gonadotropin results in maturation of the gland and milk production, accompanied  by the immirgration of lymphocytes and their activati-n to Ig formation, predominantly of the IgA class. In the presence of antigen-induced intraocular inflammation, the activated gland is able to influence the Ig class of B cells in the neighboring ocular tissues. These data suggest that even nonlymphoid tissues may elaborate lymphocyte-homing and polyclonal B-cell activating factors which function independently of specific antigen.


Disease associations of the Ia-like human alloantigens. Contrasting patterns in rheumatoid arthritis and systemic lupus erythematosus. Increasing evidence has been obtained of the special value of Ia-like B-cell alloantisera for demonstrating disease associations with histocompatibility antigens. This was particularly evident for the study of the immunogenetics of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), two conditions frequently considered related. The profiles of antigens recognized by the alloantisera in patients from each disease group was distinctive. Two types of alloantisera were obtained that illustrated the divergence between the twod iseases. One type showed a higher than normal incidence in RA but lower than normal in SLE; the other showed a higher incidence in SLE. While these sera were not totally defined, evidence was obtained that the SLE-reactive alloantiserum related to two alleles of the major histocompatibility complex DRw2 and DRw3, while the RA-reactive alloantiserum related to a common specificity shared by cells positive for either DRw4, DRw7, or DRw10. The data indicate that immunogenetic factors are relevant to the development of both RA and SLE, but that these are distinct for each disease.


The problem-oriented unit record in residency management. Application of the "medical model" of supervision to a university affiliated residency network resulted in a problem-oriented record for each unit. A data-base tabulation and standardized problem list were developed. These have been found useful in the supervision/consultation process for established programs, in the assistance of developers of new programs, and in evaluative reporting.


The sarcomere length-tension relation in skeletal muscle. Tension development during isometric tetani in single fibers of frog semitendinosus muscle occurs in three phases: (a) in initial fast-rise phase; (b) a slow-rise phase; and (c) a plateau, which lasts greater than 10 s. The slow-rise phase has previously been assumed to rise out of a progressive increase of sarcomere length dispersion along the fiber (Gordon et al. 1966. J. Physiol. [Lond.]. 184:143--169;184:170--192). Consequently, the "true" tetanic tension has been considered to be the one existing before the onset of the slow-rise phase; this is obtained by extrapolating the slowly rising tension back to the start of the tetanus. In the study by Gordon et al. (1966. J. Physiol. [Lond.] 184:170--192), as well as in the present study, the relation between this extrapolated tension and sarcomere length gave the familiar linear descending limb of the length-tension relation. We tested the assumption that the slow rise of tension was due to a progressive increase in sarcomere length dispersion. During the fast rise, the slow rise, and the plateau of tension, the sarcomere length dispersion at any area along the muscle was less than 4% of the average sarcomere length. Therefore, a progressive increase of sarcomere length dispersion during contraction appears unable to account for the slow rise of tetanic tension. A sarcomere length-tension relation was constructed from the levels of tension and sarcomere length measured during the plateau. Tension was independent of sarcomere length between 1.9 and 2.6 microgram, and declined to 50% maximal at 3.4 microgram. This result is difficult to reconcile with the cross-bridge model of force generation.


Mortality and psychosis in a peasant society. A survey was conducted of psychotic persons in a peasant society where psychiatric services were not available. Their age distribution and duration of psychosis indicated that they had a reduced longevity as compared to the general population. Intensive study of six fatal and near fatal cases suggested that violence and infection accounted for their early demise. Those with organic psychosis appeared to have a greater mortality than those with functional psychosis. As our social policies toward the mentally ill change, these findings may have applicability in our own society.


Clinical effects of aspirin and acetaminophen on hemostasis after exodontics. The effects of aspirin and acetaminophen on postextraction hemostasis were evaluated in a double-blind study in a group of 43 patients; 20 received aspirin and 23 received acetaminophen. No significant difference was found in the observed postsurgical alveolus bleeding times or the patients' reported length of postoperative bleeding. However, Ivy bleeding times taken after the ingestion of 80 grains of medication showed an elevation in 75% of the aspirin users and in 60% of the acetaminophen group; the mean change in bleeding times was 2.75 times greater in the aspirin group.


Problems involved in grading the severity of hemophilia by test results. Some problems involved in grading the severity of hemophilia were discussed. No definite correlation existed between the clinical severity of hemophilia and the results of coagulation tests (activity of deficient factors, whole blood clotting time, thromboplastin screening test). There was also no correlation between the presence or absence of oral bleeding or hemarthrosis and the test results. It was concluded that the clinical severity of the disease can be graded more reasonably on the basis of clinical hemorrhagic symptoms than according to the activity of deficient factors.


Heterotransplantation of human neoplasms. The heterotransplantation of 39 solid human neoplasms into athymic and asplenic-athymic mice resulted in the successful growth of 28 of the transplants. However, only 17 of the tumors could be maintained in serial passage. Metastasis or local infiltration by the transplanted tumors was not observed. Melanomas and colorectal carcinomas grew well whereas other types grew poorly or not at all. Possible factors responsible for the poor growth or lack of growth are discussed. The rapid growth of a melanoma in newborn mice in contrast with the slow growth of the same tumor in adult mice suggests a new experimental approach for transplantation of malignancies which previously have been difficult to establish.


Quantitation of the anti-inflammatory agent fenbufen and its metabolites in human serum and urine using high-pressure liquid chromatography. Specific procedures are described for the determination of fenbufen and its metabolites in serum and urine using high-pressure liquid chromatography. Serum or urine extracts were chromatographed on a bounded reversed-phase partitioning column. The sensitivity of the assay for fenbufen was 0.5 microgram/ml in serum with 2-ml samples and 1.0 microgram/ml in urine with 1-ml samples. The procedures are suitable for bioavailability and pharmacokinetic studies.


Fever and survival in rabbits infected with Pasteurella multocida. 1. Fever and survival rate of New Zealand white rabbits, injected with two doses of live Pasteurella multocida, were compared to determine if the relation between fever and survival observed in reptiles is also seen in a mammal. Since it is known that fever is beneficial in infected reptiles, our experiments were viewed as an initial step in the investigation of a similar potentially beneficial effect in mammals.2. There was a statistically significant correlation between the fever magnitude and survival. As fever increased up to 2.25 degrees C, the survival rate increased. Fevers above this level were associated with a decreased survival rate.3. Antipyretic drugs were administered to half the rabbits. The drugs were ineffective in reducing the fevers produced by injections of large doses of bacteria. Rabbits infused with antipyretics had a decreased mortality rate. The decreased mortality rate may be due to some action of the drugs other than their antipyretic effect.4. In vitro growth of P. multocida at normal (39 degrees C) and febrile (42 degrees C) temperatures was not significantly different. This suggests that the increased survival rate associated with higher fevers was not due to a direct inhibition of the growth of the bacteria by the increased temperature. It may be due to an enhancement of some aspect of the rabbits' immunological defences.5. It remains to be determined whether fever causes a higher survival rate or whether fever is only correlated with survival rate.


T-tubule swelling in hypertonic solutions: a freeze substitution study. Striated muscles from Rana pipiens have been exposed for variable periods of time to Ringer solutions made hypertonic by addition of either sucrose or sodium chloride. The muscles have been rapid-frozen and then prepared for electron microscopy by either freeze-substitution, freeze-fracture or cryoultramicrotomy. The only compartment greatly affected by hypertonicity is the transverse tubular system, which is visibly swollen. None of the elements of the sarcoplasmic reticulum increase in size.


Inward calcium current in twitch muscle fibres of the frog. 1. Voltage clamp experiments using the three micro-electrode voltage clamp technique were performed on sartorius muscles of the frog. 2. By blocking potassium currents with tetraethylammonium and replacing chloride ions with sulphate a slow inward current was detected. 3. The slow inward current is mainly carried by calcium, since it is abolished by cobalt and D-600, it depends on external calcium, and is not affected by removing external sodium or by tetrodotoxin (TTX). 4. The slow inward current has a mean threshold of -40 mV, reaches a mean maximum value at ca. 0 mV of 81 microamperemetercm-2 and has a mean reversal potential of +38 mV. 5. The calcium current is inactivated by the application of 2 sec conditioning prepulses according to a sigmoid curve with V(h) = -42 mV and k = 6.2 mV. 6. The slow time course of this calcium current makes it rather unlikely that it participates in contraction during a twitch, but it might be activated during long depolarizations as potassium contractures.


An analysis of the cable properties of frog ventricular myocardium. 1. The passive and active electrical parameters of frog ventricular myocardium have been measured. 2. The cytoplasmic resistivity has been determined by following changes in the resistance of a micro-electrode on penetration of a cell. 3. Unidimensional cable analysis using direct and alternating currents revealed the presence of a single time constant attributed to the surface membrane. 4. Longitudinal impedance measurements indicate that a second time constant is present in the intracellular pathway. 5. The results indicate that the resistance between cells is low so that action potentials can propagate from cell to cell by local circuits. 6. A three-dimensional cable analysis has also been carried out and compared to a simplified mathematical model which is presented in an Appendix and which closely approximates the experimental situation.


A comparison of current-voltage relations for full and partial agonists. 1. Local conductance changes produced by various bath-applied agonists at frog end-plate membrane were measured using focal recording of extracellular potential in voltage-clamped muscle fibres. The potential difference between a focal micropipette placed on the nerve terminal and another micro-pipette placed on or near inactive membrane was taken as proportional to the agonist-induced current through a small patch of an end-plate membrane. 2. The current-voltage (I--V) relation of active membrane was obtained directly by increasing the membrane potential in a ramp fashion. The change in membrane potential was slow enough for post-synaptic gating processes to reach equilibrium during the ramp. 3. During application of sufficiently low concentrations of full agonists (carbachol, (ACh) and partial agonists (choline and decamethonium) the I--V relation of end-plate membrane showed strong curvature in the range of -60 to -130 mV. The slope of I--V relations increased exponentially with membrane hyperpolarization, an e-fold change in conductance occurring for about 50 mV potential shift. 4. The curvature of the I--V relation of end-plate-membrane activated by the partial agonists choline and decamethonium became less as the agonist concentration was increased, and with high concentrations (choline 15 mM; decamethonium 250 micrometer) the I--V relation became almost straight. 5. When end-plate currents produced by high concentrations of partial agonists were matched by application of equi-active concentrations of carbachol, the carbachol-activated membrane still showed as much curvature in its I--V relation as when low concentrations of carbachol were used. 6. Choline and decamethonium concentrations for which the I--V relation was straight produced much greater depression of miniature end-plate currents than did carbachol concentrations which produced the same membrane current at the holding potential. 7. I--V relations for full agonists at high concentrations were obtained after alpha-bungarotoxin pre-treatment. During application of carbachol (400--500 micrometer) and ACh (30--40 micrometer; after complete inhibition of acetylcholinesterase activity) the I--V relation of end-plate membrane is much less curved than during application of low concentrations. 8. It is concluded that either the voltage sensitivity of agonist-induced end-plate conductance reflects voltage sensitivity of agonist binding, or the partial agonists used can exert a voltage-dependent 'local anaesthetic' action in addition to their agonist activity.


The effect of vasectomy on the testes of inbred Lewis rats. By 3 months after surgery, testicular weights were 12-14% below control values in a large series of vasectomized rats of the inbred Lewis strain. Testicular sperm concentrations in the vasectomized rats were 25-29% below those in the sham-operated animals. Although modest in magnitude, these differences were highly significant (P less than 0.001). Histometric analysis of testes from a smaller series of vasectomized Lewis rats revealed various proportions of abnormal seminiferous tubules. The degenerative changes included reduction in tubule diameter, fusion of spermatids, and depletion of advanced germinal elements. In the most severely affected tubules, leakiness of the blood-testis barrier was indicated by penetration of lanthanum through Sertoli-cell tight junctions.


Synthesis and antiinflammatory activity of 6,11-dihydro-11-oxodibenzo[b,e]thiepinalkanoic acids and related compounds. Acetic acid derivatives of tricyclic systems, such as 6,11-dihydro-11-oxodibenzo[b,e]thiepin, 4,10-dihydro-4-oxo-thieno[2,3-c][1]benzothiepin, dibenzo[b,f]thiepin, dibenz[b,f]oxepin, etc., were synthesized and assayed for antiinflammatory activity. One of the compounds, 6,11-dihydro-11-oxodibenzo[b,e]thiepin-3-acetic acid (52), was chosen for evaluation in man on the basis of high antiinflammatory activity in both short- and long-term animal assays and a low gastric irritation liability in rats and dogs.


Novel analgesic-antiinflammatory salicylates. 5-(2,4-Difluorophenyl)salicylic acid, diflunisal (25), is the best compound, in terms of both efficacy and safety, from over 500 salicylates investigated in our laboratories. It is a chemically distinct, nonacetylating salicylic acid, more active than aspirin as an analgesic and antiinflammatory agent and superior in duration of action and therapeutic index. Some recent clinical and biochemical observations are briefly discussed.


Synthesis and analgesic-antiinflammatory activity of some 4- and 5-substituted heteroarylsalicylic acids. We have made a series of 4- and 5-aryl- and 4- and 5-heteroarylsalicylic acid derivatives with the objective of reducing gastric irritation and increasing potency. Here we describe a series of 4- and 5-heterocyclic salicylic acids and their antiinflammatory-analgesic potencies measured in comparison to aspirin. An improvement of the therapeutic index over aspirin of 100 was achieved; however, the heterocyclic salicylic acids lacked antipyretic activity. Some physicochemical parameters which may bear on the antiinflammatory activity of these compounds are discussed.


Antiinflammatory and analgesic diastereoisomeric derivatives of indan-5-acetic acid. Various 2-alkyl-alpha-methyl- and 2-alkylindan-5-acetic acids have been prepared. The acids, which can exist in two diastereoisomeric forms that cannot be separated by crystallization or chromatography, can be analyzed in their mixture by NMR in the presence of Eu(dpm)3. It has been possible to reconstitute the two pure racemic 2-isopropyl-alpha-methylindan-5-acetic acids from their enantiomers obtained after resolution of the mixtures through salts with various active bases. The relative configuration of the two asymmetric centers of one of the diastereoisomers salts with various active bases. The relative configuration of the two asymmetric centers of one of the diastereoisomers has been determined by X-ray crystallography. The absolute configurations of the resolved acids have been established by a comparative study of their CD curves. The antiinflammatory and analgesic properties of these compounds as functions of their structure and stereochemistry are discussed.