Stage-sensitivity and dose-response study after gamma-irradiation of mouse primary spermatocytes. The sensitivity pattern for X-ray-induced chromatid aberrations was analysed in primary spermatocytes irradiated at 1 to 11 days diakinesis-metaphase I of meiosis. Using a dose of 300 R (60 R/min) of gamma-rays zygotene (day 9) was found to be the most sensitive and leptotene (day 11) the most insensitive stage. The dose-response to gamma-ray-induced aberrations was evaluated in cells irradiated zygotene. Both the yields of rearrangement as well as the yields of fragments gave a best fit to a quadratic model.


Arterial wall damage by X-rays and fast neutrons. Radiation-induced atheromatosis has been studied for 200 kVp X-rays and 15 MeV neutrons. From the results of two earlier experiments, a r.b.e. less than 1 was expected for neutrons. (1) Irradiation of blood-vessels causes depolymerization of the mucopolysaccharides in the vessel wall, resulting in atheromatosis, and (2) in two other mucopolysaccharide systems a r.b.e. less than 1 is observed for fast neutrons. Irradiation of the carotid arteries of a total number of 120 hypercholesterolaemic rabbits, divided over several groups, with 500 and 1000 rad of X-rays or neutrons results, however, in atheromatous plaques which are more pronounced for neutrons than for X-rays at the 500 rad dose-level; for a dose of 1000 rad the effect of neutrons is less extensive than the effect of X-rays. These results lead to the assumption that the origin of the atheromatosis seems not only to be the mucopolysaccharide depolymerization, but that radiation induced damage at the cellular level must be taken into account.


O2 consumption of the amphibian cornea: relationship to Na+ and Cl- and the presence of drugs. Thirty percent of the O2 consumption of the amphibian cornea was Na-dependent. Ouabain, in the presence but not the absence of this cation, inhibits O2 uptake by about 30 patient. In contrast, neither the presence of Cl- nor drugs which stimulate (adrenaline, theophylline) or inhibit (furosemide, thiocyanate) active transport of this anion had detectable effects on O2 consumption.


Small-angle light-scattering patterns of corneas of different species. Light-scattering patterns of corneas of different species have been obtained. The different small-angle light-scattering (SALS) patterns were classified into four groups on the basis of the angular dependence of the intensity of light scattered in the I+ and I modes. At present, only two types of patterns can be explained on the basis of theoretical models. The need to develop a general model that can account for all four types of SALS patterns is discussed.


Polyene antibiotics. IX. An improved method for the preparation of methyl esters of polyene antibiotics. An improved general method for the preparation of methyl esters of polyene antibiotics is discussed. Using this method methyl esters of pimaricin, nystatin, eurocidin, hamycin, hamycins A and B, aureofungin, partricins A and B, candimycin, candicidin, and amphotericin B have been prepared and their physical properties are reported. The biological activities of hamycins A and B and their methyl esters are also described.


Purification and properties of colony-stimulating factor from mouse lung-conditioned medium. Colony-stimulating factor, which specifically stimulates mouse bone marrow cells to proliferate in vitro and generate colonies of granulocytes, or macrophages, or both, was purified 3500-fold from mouse lung-conditioned medium. Analysis by discontinuous polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate indicated that there was a single protein component. All of the colony-stimulating activity was coincident with the protein band. The molecular weight of colony-stimulating factor estimated by gel filtration was approximately 29,000 and by electrophoresis approximately 23,000. The specific activity of purified colony-stimulating factor from mouse lung-conditioned medium bound to concanavalin A-Sapharose, indicating that it is a glycoprotein. The small percentage of colony-stimulating factor in mouse lung-conditioned medium which did not bind to concanavalin A-Sepharose appeared to represent molecules which lacked the carbohydrate moieties required for binding to this lectin. It was necessary to include low concentrations (less than 0.01%, v/v) of polymers such as gelatin and polyethylene glycol, or nonionic detergents such as Triton X-100, in all of the buffers used throughout the purification scheme, otherwise colony-stimulating factor was lost from solution. At high concentrations (greater than 20 mug/ml) the factor stimulated the formation of granulocytic, macrophage, and mixed colonies from C57BL mouse bone marrow cells. As the concentration of purified colony-stimulating factor was decreased, the frequency of colonies containing granulocytes also decreased. At low concentrations of colony-stimulating factor (less than 70 pg/ml) only macrophage colonies were stimulated.


Quaternary structure of delta-aminolevulinic acid synthase from Rhodopseudomonas spheroides. Delta-Aminolevulinic acid synthase (succinyl-CoA: glycine C-succinyltransferase (decarboxylating) EC 2.3.1.37) was purified from Rhodopseudomonas spheroides. The purity of the enzyme preparation was established by its behavior in disc electrophoresis in the presence and absence of sodium dodecyl sulfate and by analytical ultracentrifugation. The molecular weight of the enzyme as determined by sedimentation equilibrium was found to be about 80,300, a value similar to those obtained by gel filtration, polyacrylamide gel electrophoresis, and sucrose gradient centrifugation. The molecular weight of the enzyme, denatured with either sodium dodecyl sulfate or guanidine hydrochloride, was found to be about 45,000 and 41,000, respectively. The dimeric structure was supported by sedimentation in sucrose gradients. Further evidence for the dimetic nature of the enzyme was obtained by gel electrophoresis of the enzyme treated with dimethylsuberimidate and sodium dodecyl sulfate.


Effects of fibroblast growth factor and epidermal growth factor on the rate of growth of amniotic fluid-derived cells. Primary cultures of cells derived from human and bovine amniotic fluid were cultivated in Dulbecco's modified Eagles medium with 20% fetal calf serum. Whereas increased concentrations of fetal calf or other types of serum bring about no further mitogenic response, the rate of growth of these cells was nevertheless sharply increased when 100 ng/ml of FGF was added to the culture media. The FGF induced mitogenic effect was statistically significant. EGF at 100 ng/ml had a less pronounced effect which, when analyzed statistically, was not significant. Amniotic fluid-derived cells are used for the prenatal detection of genetic disorders. The use of FGF to increase the rate of growth of these cells should reduce the time required between amniocentesis and diagnosis.


Modification of factor VIII complex properties in patients with liver disease. Factor VIII procoagulant activity (VIII:C), factor VIII related antigen (VIIIR:AG), and the von Willebrand factor (VIII:WF) were measured in 19 patients with liver disease (8 cases of alcoholic cirrhosis and 11 with acute viral hepatitis). In both groups of patients the levels of VIII:C, VIIIR:AG, and VIIIR:WF were above normal or normal with mean values well above the normal range. VIIR:AG was also measured in the same patients by two-dimensional electrophoresis, and its biological properties were measured after purification by chromatography. By both methods the VIIIR: AG in the patients with liver disease differed from normal VIIIR:AG. In five of the patients with acute viral hepatitis, who were tested after they had recovered, the previously elevated levels of VIII:C, VIIIR:AG, and VIIIR:WF had dropped to normal limits and the qualitative abnormalities had disappeared.


Palindromic rheumatism: an allergic manifestation? A case of palindromic rheumatism is presented in which the arthritic attacks usually were accompanied by an allergic vasculitis and dermatitis. Although this is not proof of the etiology, the constant association is suggestive of an allergic cause. A brief review of palindromic rheumatism in included.


Receptors for IgM-coated erythrocytes on chronic lymphatic leukemia cells. Our experiments show that lymphocytes of CLL patients, having typical B cell characteristics, form rosettes with IgM-coated bovine erythrocytes. Of 18 investigated patients, 3 to 78% (mean 29%) of the isolated lymphocytes reacted with EA-IgM. With mixed rosette assays. EA-IgM bound to cells bearing receptors for IgG as well, but not receptor-bearing lymphocytes. Rosette formation could be completely blocked by addition of IgM at concentrations as low as 0.17 mg/ml. Ten milligrams per milliliter of aggregated human IgG had no effect on the rosette formation with EA-IgM but completely abolished the binding of EA-IgG. Adult human or rabbit serum blocked the EA-IgM binding, whereas cord blood serum and FCS had no effect. These inhibition data indicate that EA-IgM binding does not occur via a somewhat altered IgG-Fc receptor but reacts with different membrane structures. That EA-IgM receptor can be cleaved off with trypsin and can be reconstituted after overnight cultivation, also supports this viewpoint. In contrast to the situation in normal subjects, in CLL patients the IgM receptors are demonstrable before overnight cultivation and are found on cells with B cell characteristics.


T and B lymphocyte migration into syngeneic tumors. The migration of splenic T and B lymphocytes into syngeneic tumors undergoing immunologic rejection was investigates. Spleen cells were obtained from normal BALC/c mice or BALB/c mice bearing tumors induced by murine sarcoma virus (MSV). Either whole spleen cells or immunoabsorbent purified T and B cells were radiolabeled with sodium chromate-51 and injected i.v. into normal or MSV inducted-tumor bearing syngeneic recipients. Twenty-four hours later the recipient mice were sacrificed and radioactivity was assessed for tumor, contralateral normal muscle, the lymph nodes draining the tumor and contralateral draining lymph nodes, peripheral lymph nodes, spleen, and liver. Both T and B lymphocytes from either normal or MSV tumor-bearing animals show greatly increased migration into the tumor when compared with normal muscle. Migration of T cells from both normal and MSV tumor bearers was 30 times that of migration to normal muscle. B cells from tumor-bearing mice, on the other hand, localized in the tumor itself only 50% as frequently as did B cells from normal animals. In addition, T cells from MSV tumor bearers were found in the highest proportion in the lymph node draining the tumor site. We conclude that T and B lymphocytes from either normal or tumor-bearing mice migrate to a syngeneic tumor undergoing immunologic rejection. In contrast, the migration of both T and B cells from tumor-bearing animals was decreased to the peripheral lymph nodes at the time of maximum tumor growth.


Antigen-initiated B lymphocyte differentiation. IX. Characterization of memory AFC progenitors by buoyant density and sedimentation velocity separation. The characteristics of memory B cell antibody-forming cell (AFC) progenitors from long-term hapten-primed CBA mice were investigated by using sedimentation velocity and buoyant density separation to isolate physically distinct B cell sub-sets. The isolated fractions were assayed by the adoptive immune response to NIP-POL antigen, under conditions where neither T cells nor other accessory cells were limiting the IgM or IgG AFC responses. The results were compared to previous studies on the IgM AFC-progenitors of unprimed adult mice. Splenic IgM and IgG memory AFC-progenitor activity was largely found among the typical B cells of slow to medium sedimentation rate, in contrast to the fastre sedimenting IgM AFC-progenitor activity of unprimed animals. Splenic IgM and IgG memory AFC-progenitor activity was found among the medium to light density cells, and so resembled by this parameter the IgM AFC-progenitor activity in unprimed animals. Thoracic duct lymphocytes from hapten-primed mice also exhibited memory IgM and IgG AFC-progenitor activity in the slow-medium sedimentation range. However, in contrast to spleen, the IgM and IgG memory AFC-progenitor activity in lymph was found among very dense B cells. Two physically distinct sub-populations of memory B cells have thus been identified, namely: i) small, medium-light density, presumably tissue-resident B lymphocytes found in spleen; and ii) small, dense, presumably recirculating B lymphocytes found in lymph. Both physical forms include IgM and IgG progenitors. Both forms are distinct from the larger, medium-light density "virgin" AFC-progenitors in the spleen of unprimed adult mice.


Differentiation of B lymphocytes in C3H/HeJ mice: the induction of Ia antigens by lipopolysaccharide. The lipid A moiety of bacterial lipopolysaccharide (LPS) elicits several types of responses in murine B lymphocytes. First, lipid A induces the nonproliferative expression of cell surface antigens in more immature cell types. Second, lipid A induces a mitogenic response in more mature B cell types. Lipid A induces the expression of Ia antigens on bone marrow cells from C3H/DiSn but not C3H/HeJ mice. The Ia-inducible cells possess surface immunoglobulin. Agents that elevate intracellular levels of adenosine 3',5'-cyclic monophosphate (cyclic AMP) induce the appearance of Ia antigens on B lymphocytes from both C3H/HeJ and C3H/DiSn mice, suggesting that lipid A exerts its inductive effects by increasing cyclic AMP levels in cells. In contrast to what is observed by using other strains of mice, mature B lymphocytes from C3H/HeJ mice do not support a mitogenic response to lipid A. The subpopulation of B lymphocytes in C3H/HeJ mice that normally respond mitogenically to LPS not only appear to lack an LPS-response mechanism utilized in the mitogenic pathway, but they lack the LPS-response pathway of the immature B cell types. A lipid A-bound protein (LAP) induces both the expression of Ia and a mitogenic response in the different subpopulations of B lymphocytes from C3H/HeJ and C3H/DiSn mice. The genetic defect in C3H/HeJ mice that limits responses to lipid A may be associated with a receptor that is normally expressed on many different cell types.


Fractionation of lymphocytes involved in the generation of cell-mediated cytotoxicity over insolubilized conjugated histamine columns. Spleen cells from normal BALB/c mice were cultured in vitro with irradiated C57BL/6 stimulating cells. Five days later the T cell-mediated cytotoxic activity of the effector cells was assessed with a 51Cr-release assay that used H-2bEL-4 tumor cells as targets. Before the BALB/c responding lymphocytes were sensitized they were fractionated by passing the spleen cells over insolubilized histamine rabbit serum albumin Sepharose columns (H-RSA-S) or over rabbit serum albumin Sepharose (RSA-S) control columns. Fractionation of cells over the H-RSA-S columns depleted or significantly reduced the cytotoxic potential of the unretained cells. All cytotoxic potential was recovered when the cells that adhered to the H-RSA-S were eluted from the columns. In contrast, no effect on responsiveness was detected after the cells had been fractionated over the control column. The loss of response potential by the cells that did not adhere to H-RSA-S could not be accounted for by removal of macrophages nor by the concentration of cells with suppressor activity in the effluent. These cell fractionation studies raise the possiblity but do not prove that cytotoxic precursor cells may express amine receptors that could be responsible for their retention by insolubilized histamine columns.


Analysis of the major histocompatibility complex in Syrian hamsters. III. Cellular and humoral immunity to alloantigens. Among the genetic loci incorporated into the major histocompatibility complex in every species studied to date have been prominent genes encoding for strong histocompatibility determinants that elicit detectable alloantibody responses and which are the chief antigenic targets of cell-mediated cytotoxicity reactions. The K and D regions of the H-2 complex in the mouse and the A, B, and C regions of the HLA complex in man are representative examples. Syrian hamsters, as described in this report, do not make alloantibodies to antigens of this type and only very poorly do they carry out in vitro cell-mediated cytotoxicity to target cells putatively bearing these antigens. Since hamsters are quite capable of discriminating analogous antigenic differences in xenogeneic species, and xenogeneic sources cannot distinguish immunologically between the antigens encoded by the two hamster major histocompatibility alleles. Hm-1a and Hm-1b, we conclude that the hamster strains we work with are serologically indistinguishable by the methods used here. However, they obviously differ for determinants which elicit T cell-mediated responses, as evidenced by their ability to express acute skin graft rejection, mixed lymphocyte reactivity, graft-vs-host reactions, and cell-mediated cytotoxicity reactions. Such alloreactivity may reflect a mutation at an SD locus, affecting antigenic sites recognized only by T cells, or that the available hamster strains are SD identical, but differ at loci similar to the I region loci in mice. Alternatively, we cannot exclude the possibility that Syrian hamsters somehow fail to express properly the genes coding for SD determinants.


Regulation of antibody response in different immunoglobulin classes. II. Induction of in vitro IgE antibody response in murine spleen cells and demonstration of a possible involvement of distinct T-helper cells in IgE and IgG antibody responses. In vitro induction of anti-DNP IgE as well as IgG1, IgG2a antibody responses was shown in murine spleen cell culture. Spleen cells primed three times with 1 mug of DNP-OA or DNP-Asc produced significant amounts of anti-DNP IgE as well as IgG antibodies by the in vitro stimulation with DNP-OA or DNP-Asc, respectively. Collaboration between DNP-primed B cells and carrier-primed T cells was required for the induction of both IgE and IgG antibodies with DNP-coupled T-dependent antigen. Carrier-specific T cells induced with a low dose of Asc (0.01 mug) showed helper function only on IgE antibody response, whereas T cells primed with a higher dose of Asc (10 mug) cooperated only with IgG-B cells. T cells primed with Asc in CFA showed helper function mainly on IgG antibody response but not on IgE antibody response. The result indicated the presence of a distinct population of T helper cells for IgE and IgG antibody responses. T-independent antigen (DNP-Ficoll) induced both anti-DNP IgE and IgG antibody responses in DNP-primed spleen cell population without the requirement of the collaboration of helper T cells.


Age-related decrease in mouse T cell progenitors. Mice were given a lethal dose of whole-body gamma-radiation and injected with a 10(5) or 10(6) marrow cells from 10- to 143-week-old syngeneic donors. Nine days later, colony-forming units (CFU) were counted in the spleens of mice given 10(5) cells, and 15 to 21 days after irradiation thymus weights and in some experiments 3H-thymidine uptake or total thymic cellularity were determined in the recipients of 10(6) cells. It was found that in the majority of mouse strains studied there were no significant changes with age in marrow CFU. In contrast, thymic regeneration was significantly impaired when the recipients received marrow cells from donors 100 weeks of age or older. These observations and results obtained in dose-response and time-course studies are best explained by an age-related decrease in marrow T cell progenitors; however, certain findings suggest that in addition the proliferative capacity of these stem cells may at times be moderately impaired.


Cross-linking of T cell mitogens: effects on B and T cell proliferation and immunoglobulin synthesis. The T cell mitogens Pa-2, concanavalin A (con A) and its dimeric derivative succinyl-con A, were each cross-linked with the bifunctional reagent dimethyl suberimidate. Although the dose-response curves of these insoluble aggregated products were markedly changed from those of the soluble mitogens, each aggregate continued to stimulate DNA synthesis by murine thymus and T cells. Both aggregated Pa-2 and aggregated succinyl con A stimulated DNA synthesis by B cells from athymic (Nu/Nu) mice. Aggregated con A did not stimulate these cells and, like soluble con A, depressed the background incorporation of 3H-thymidine. Unlike soluble Pa-2, aggregated Pa-2 also greatly increased Ig production by both the B cell cultures and B + T cell cultures from normal (BALB/c) mice.


Distribution of alloantigens on human Fc receptor-bearing lymphocytes: the presence of B cell alloantigens on sIg-positive but not sIg-negative lymphocytes. Human peripheral blood lymphocyte subpopulations were analyzed for the presence of B cell alloantigens with a microcytotoxicity assay. B cell alloantigens were found exclusively on sIg-positive lymphocytes and were not present on sIg-negative, Fc receptor-bearing lymphocytes or sIg-negative, Fc receptor-negative T lymphocytes.


Induction of cell-mediated immunity to chemically modified antigens in guinea pigs. II. The interaction between lipid-conjugated antigens, macrophages, and T lymphocytes. Protein antigens covalently conjugated with lipid groups (dodecanoic acid) have previously been shown to stimulate strong delayed-type hypersensitivity (DTH) without the aid of adjuvants. The present experiments show that lipid-conjugated bovine serum albumin (L-BSA) is taken up in vitro by macrophages (Mpsi) 25- to 50-fold more than unconjugated BSA or aminidated BSA, neither of which induces DTH. Macrophages that take up 125I-labeled L-BSA in vitro stimulate DTH even more efficiently, when injected into syngeneic guinea pigs, than does soluble L-BSA. Tracer studies on the fate of radiolabeled BSA and L-BSA showed that much more L-BSA than BSA was retained by draining lymph nodes. Autoradiography demonstrated that 125I-L-BSA is rapidly taken up by Mpsi in the medullary sinuses of the lymph nodes. Some of this antigen is then transported into the paracortex, a region in which T lymphocytes predominate. The capacity of lipophilic antigens to stimulate cell-mediated immune responses may be caused by their increased uptake by Mpsi, resulting in more efficient presentation to immunocompetent T lymphocytes. The anatomical site of this Mpsi-T cell interaction may be within the sinusoids or paracortex of the draining lymph nodes.


In vitro induction of tumor-specific immunity. III. Lack of requirement for H-2 compatibility in lysis of tumor targets by T cells activated in vitro to oncofetal and plasmacytoma antigens. Many recent studies have demonstrated that cytotoxic T lymphocytes (CL) activated to various antigens other than those of the H-2 complex, will lyse target cells only when H-2 compatibility exists between the CL and target cell. From these observations, it has been inferred that T lymphocytes might only be capable of responding to H-2 antigen or antigens that become associated with H-2 region gene products. Our results suggest that this is not the case, and that in some situations, cytotoxic T lymphocytes can specifically lyse target cells of different H-2 types. Two in vitro systems are described where primary induction of cytotoxic T lymphocytes to oncofetal and plasmacytoma antigens results in CL capable of lysing suitable targets bearing these antigens, of either syngeneic or allogeneic derivation. Thus it is proposed that although interaction antigens involving H-2 components may preferentially activate T lymphocytes, this does not imply a restriction on the recognition potential of T lymphocytes.


Inhibition of antibody-dependent cellular cytotoxicity by protein A from Staphylococcus aureus. Protein A, a cell wall constituent of several strains of Staphylococcus aureus, binds strongly to the Fc portion of immunoglobulins. This investigation demonstrated that such binding can inhibit antibody-dependent cellular cytotoxicity (ADCC). The degree to which ADCC was inhibited depended upon the relative concentrations of protein A and anti-target cell antiserum. Protein A also inhibited the formation of rosettes between antibody-coated sheep red blood cells and lymphoid cells with Fc receptors. We, therefore, conclude that protein A inhibits ADCC by preventing the binding of antibody-coated target cells to Fc receptors on cytotoxic effector cells.


Functional characteristics of Peyer's patch lymphoid cells. IV. Effect of antigen feeding on the frequency of antigen-specific B cells. The frequency of B cells in Peyer's patches from normal BDF(1) and sheep red blood cell (SRBC)-fed BDF(1) mice that could respond to antigenic determinants on SRBC and trinitrophenyl (TNP) was determined using an in vitro system of limiting dilution analysis. In normal mice, one B cell in 1.9 x 10(4) Peyer's patch cells could be induced to an anti-SRBC response and one B cell in 3.6 x 10(4) Peyer's patch cells could be induced to an anti-TNP response. The frequency of B cells capable of responding to SRBC in normal mice was similar in Peyer's patches and spleen. However, after feeding mice SRBC for 3 weeks, there was a 6-fold reduction in the frequency of B cells in Peyer's patches capable of responding to SRBC but no change in the frequency of B cells capable of responding to TNP. The average clone size of Peyer's patch B cells responding to SRBC was similar in normal and SRBC-fed mice. Although antigen-feeding does not stimulate Peyer's patch B cells in situ to humoral antibody synthesis, antigen-feeding can markedly alter the reactivity of the antigen-sensitive cell population in Peyer's patches. We previously demonstrated that T cells in Peyer's patches could be specifically carrier primed for helper function by SRBC feeding. We have now demonstrated that antigen-feeding reduced significantly the frequency of B cells in Peyer's patches capable of responding to the fed antigen. Peyer's patches appear to serve an important function as a sampling site for intestinal antigens.


Interactions between human lymphocytes and paramyxovirus-infected cells: adsorption and cytotoxicity. The capacity of human lymphocytes to adhere to paramyxovirus-infected monolayers and their capacity to kill paramyxovirus-infected cells was investigated. A large fraction of human lymphocytes was found to adhere firmly to the paramyxovirus-infected monolayers. Predsorption of lymphocytes on mumps virus-infected cells impaired their  adsorption to a second cell monolayer of the same type. The cytotoxic activity of lymphocytes against mumps virus-infected cells was also reduced after predsorption on mumps virus- or Newcastle disease virus-infected (NDV) cell monolayers. Exposure of lymphocytes to trypsin did not significantly decrease either adsorption or cytotoxicity. Pretreatment of lymphocytes with neuraminidase (NANase) partly inhibited adsorption whereas cytotoxicity was not decreased. Cell fractionation experiments after rosetting of the lymphocytes with sheep erythrocytes (E) indicated that T cells were equally or better adsorbed than "non-T" cells. Taken together with previous experiments which showed that the majority of T lymphocytes are not cytotoxic against mumps virus-infected cells these results suggest that adherence of lymphocytes to infected cells and cytotoxicity may be unrelated phenomena.


Recovery of soluble sheep erythrocyte receptor from the T lymphocyte surface by proteolytic cleavage. Proteolytic digestion of the human T lymphoblastoid cell line (Molt-4) and of peripheral blood lymphocytes by trypsin, chymotrypsin, and pronase results in a progressive, time-and dose-dependent diminution of T lymphocyte-sheep red bloock cell (SRBC) rosette formation, whereas thrombin, plasmin, collagenase, DNAse, and phospholipase have not effect. Complete abrogation of SRBC binding is achieved when lymphocytes (1 x 108/ml) are incubated with either trypsin or chymotrypsin at 10 mug/ml for 30 min, and greater than 50% abrogation is observed between 3 to 10 min. Preincubation of SRBC with the 10 min and 20 min lymphocyte digest supernatants inhibited their subsequent binding by normal T lymphocytes by as much as 64%. Thirty-minute digests were less inhibitory. Equivalent digests from several human B lumphoblastoid cell lines and from a non-rosetting clone of Molt-4 cells were not inhibitory. Polyacrylamide gel electrophoresis followed by elution of serial gel slices revealed four distinct inhibitory bands (I-IV) in the 20-min digest supernatant whereas only bands I-III and band IV were present in the 10-min and 30-min digest supernatants, respectively, suggesting progressive proteolysis of a distinct receptor. These experiments indicate that the binding of SRBC by human T lymphocytes represents a receptor-ligand interaction rather than a nonspecific electrical charge phe nomenon and that the receptor is a discrete molecular species which can be isolated from the surface of T but not B lymphocytes by limited enzymatic proteolysis.


Generation of cytotoxic T lymphocytes during coxsackievirus B-3 infection. I. Model and viral specificity1. The production of cytotoxic cells in the spleen of adult male BALB/c mice infected with Coxsackievirus B-3 has been examined. An in vitro 51Cr release assay was used to measure cytotoxic activity against virus-infected and uninfected neonatal sygeneic fibroblasts. Cytotoxicity of immune spleen cells against virus-infected targets was detected on the 3rd day after infection, reached a peak on day 7, and then declined to low levels by days 12 and 14. Spleen cells obtained 3 and 5 days after infection also exerted cytotoxicity against uninfected fibroblasts, but by the 7th day there was little or no reactivity against uninfected target cells, although activity against infected fibroblasts was maximal at this time. Reciprocal assays performed by using Coxsackie and vaccinia viruses provided evidence of virus specificity of the cytotoxic reaction. When spleen cells were obtained 7 days after infection, the Coxsackievirus-immune population was not cytotoxic for vaccinia-infected fibroblasts, and the vaccinia-immune population was not cytotoxic for Coxsackievirus-infected targets, although each immune cell preparation caused significant lysis of fibroblasts infected with the homologous virus. Additional studies showed that primary mouse or hyperimmune rabbit anti-Coxsackieviral serum could not block immune spleen cell cytotoxicity or induce complement-mediated lysis of infected targets. The findings indicate that Coxsackievirus infection results in surface membrane alterations, but no evidence was obtained that antiviral antibody could react with the infected cells.


Generation of cytotoxic T lymphocytes during coxsackievirus tb-3 infection. II. Characterization of effector cells and demonstration cytotoxicity against viral-infected myofibers1. This report describes studies characterizing the virus-specific cytotoxic effector cells which are present in the spleens of mice 7 days after infection with Coxsackievirus B-3. An in vitro 51Cr assay employing eyngeneic virus-infected neonatal fibroblasts was used to measure cytotoxic activity. Treatment of immune cells with (anti-thy-1.2) and complement abolished dtheir cytotoxic activity, but no reduction occurred when B cells were removed by incubation with anti-Ig and complement or macrophages eliminated by adherence depletion. The findings therefore imply that the cytotoxic reaction was mediated by sensitized T cells and that B cells and macrophages did not play an important role. Reciprocal assays performed with BALB/c and CBA/J cells showed that Coxsackievirus-immune spleen cells lysed infected syngeneic targets but not allogeneic targets, providing further evidence that cytotoxicity was mediated by effector T cells. In addition and in vitro assay system employing neonatal myocardial cells was developed and used to demonstrate that Coxsackievirus-infected myofibers were susceptible to destruction by immune spleen cells. The evidence suggests that mice infected with Coxsackie B viruses are able to mount a cell-mediated immune response with production of cytotoxic T cells which have the capacity to damage tissues infected with these agents.


Multiple occurrence of spontaneous AKR/J lymphomas with T and B cell characteristics. Three Thy 1.1-positive and surface IgM-positive (Thy 1+, SIg+) AKR/J lymphoma lines are described. These doubly marked tumors arose spontaneously in the peripheral lymphoid organs of 14- to 16-month-old AKR/J mice that either had spontaneous thymus atrophy or had been thymectomized at 1 month of age. All lines bore surface Thy 1.1, Ly,Ig(micron-chain) and Fc receptor (FcR), detectable by immunofluorescence. Immune response region (Iak) antigen was present on the two lines tested. Persistence of Thy 1.1 antigen and SIg after long-term tissue culture provided evidence that these markers were not passively acquired. One of these tumor lines, AkTB-1 always grows in lymph nodes as Thy 1.1-positive,SIg-negative tumors cells, whereas tumor cells growing in the spleen are initially Thy 1.1 positive, SIg negative, but they rapidly acquire SIg, FcR, and 1a between 18 and 21 days of passage.


Radiosensitivity of T and B lymphocytes. IV. Effect of whole body irradiation upon various lymphoid tissues and numbers of recirculating lymphocytes. Groups of 10-week-old female CBA/J mice were exposed in whole body fashion to 0,5,50, and 500 rads and sacrificed in serial fashion 1,3,5,7,9,15, and 30 days after irradiation for morphologic evaluation of thymus, spleen, lymph node, and Peyer's patch, and assessment of the relative numbers of thymus-derived (T) and bone marrow-derived (B) cells in these tissues. The absolute and relative numbers of recirculating T and B cells mobilizable by thoracic duct cannulation were also determined and compared with similar determinations with respect to peripheral blood lymphocytes. B cell depletion occurred more quickly and was more pronounced in spleen and lymph node than T cell depletion at all three exposure doses. Depletion of T and B cells was roughly equal in peripheral blood and thoracic duct lymph. When present, regeneration of the T cell component occurred more rapidly than did B cell restoration. The latter often was incomplete at the time of the final sacrifice (day 30). PHA-responsive and Con A-responsive cells also appeared to differ with respect to the kinetics of cell death after whole body irradiation.


Neonatal susceptibility to MHV3 infection in mice. I. Transfer of resistance. Up to 3 weeks of age, mice of the resistant A/J strain are fully susceptible to mouse hepatitis virus type 3 infection (MHV3). Immune deficiency, however, resulting from neonatal thymectomy or long term ALS administration led A/J animals to remain susceptible when tested at adult age. Whole spleen cells transferred from normal adult A/J donor mice protected suckling syngeneic recipients from i.p. infection with MHV3. Such a protective capacity of spleen cells was abolished after treatment with anti-theta serum and complement. Spleen cell separation by means of adherence to plastic also showed that neither the nonadherent nor the adherent populations injected separately were able to confer resistance to young mice challenged with the virus. Protection was not achieved with peritoneal cells originating from adult syngeneic animals. Transfer of resistance to MHV3 was obtained, however, when peritoneal cells were associated with adherent spleen cells. This study indicated that two types of mature cells, at least, were required for transferring MHV3 resistance into newborn mice of the A/J strain: T lymphocytes and an adherent spleen cell population.


Mechanisms of B cell tolerance. I. Tolerance to dextran B1355 induced with the oxidized dextran. The bacterial dextran B1355, which is normally a potent thymus-independent immunogen, was made tolerogenic by oxidation. The injection of the oxidized dextran into BALB/c mice before, at the same time, or up to 4 days after the injection of the immunogenic form of the dextran resulted in a marked immunologically specific suppression of the number of anti-dextran antibody-forming cells found in the spleen. This suppression resulted from a direct inactivation of antibody-forming cell precursors rather than from either inhibition of antibody secretion or the exhaustive utilization of precursor B cells that have been observed in other tolerance systems. A substantial degree of tolerance was achieved after only a 1-hr in vivo exposure of the spleen cells to the tolerogen. At a dose of 1 mg of oxidized dextran per mouse, tolerance persised for at least 3 weeks. A complete recovery was apparent by 10 weeks. The stability of the tolerance was demonstrated by transferring tolerant spleen cells to irradiated recipients. The response in the recipient animals to an immunogenic dextran challenge remained suppressed. It appears that the tolerogenicity of the oxidized dextran is due to its ability to couple covalently with free amino groups in or near the receptor site of the cell membrane via the reactive dialdehyde groups of the dextran.


Hapten-specific leukocyte migration inhibition. I. Inhibition of cells from animals immunized with DNP-KLH by epsilon-DNP-L-lysine Ficoll. Guiena pigs were immunized with dinitrophenyl-keyhole limpet hemocyanin (DNP-KLH) or with hemocyanin in complete Freud's adjevant. The migration of oil-induced peritoneal exudate cells was measured in the presence and absence of epsilon-dinitrophenyl-lysine-tyrosinyl Ficoll (DNPL-F). When the cells came from animals immunized with DNP-KLH their migration was inhibited by DNPL-F. Control cells from animals immunized with KLH or not immunized migrated normally in the presence of DNPL-F.


The quantitative production of interferon by mitogen-stimulated mouse lymphocytes as a function of age and its effect on the lymphocytes proliferative response. Interferon production by mitogen-stimulated mouse spleen cells increases as the age of the cell donor increases and also varies with with the strain of the cell donor. Exogenous interferon added to mouse spleen cell cultures at dose levels known to be produced by the cells causes a reduction in the proliferative response of T cells to mitogen stimulation. Since the spleen cells from old mice respond poorly to mitogen stimulation, it may be possible that the interferon elaborated by these cells is adversely affecting the mitogen assay.


Regulatory substances produced by lymphocytes. V. Production of inhibitor of DNA synthesis (IDS) by proliferating T lymphocytes. The conditions neccessary for production of inhibitor of DNA synthesis (IDS) by rat lymphocytes were investigated. In concanavalin A (Con A)-stimulated lymph node cell (LNC) cultures, IDS production was not  detected in the culture supernatant during the first 24 hr, and it increased gradually after that to reach a maximum at 3 to 4 days. When the cells were pretreated with mitomycin C, IDS was not produced, suggesting that DNA synthesis of LNC or a LNC subpopulation is necessary for IDS production. In contrast, Con A-stimulated spleen cells priduced a high level of IDS within 24 hr, and its production  fell off sharply thereafter. Con A-stimulated rat thymocytes also produced IDS reaching a maximum at 2 to 3 dyas. However, thymus cells from rats treated with hydrocortisone 48 hr previously did not produce IDS. This finding implies that cortisol-sensitive (cortical) thymocytes are capable of producing IDS and cortisol-resistant (medullary) thymocytes are not. IDS production by lymphoblasts was proportional to cell number and unaffected eith by cell density (1 to 10 x 106/ml) or by the concomitant presence of normal cells from spleen, lymph node, or thymus. Thus Con A-stimulated cells, after becoming blasts, appear to produce IDS automatically wihtout affecting or being affected by other cells. Both spleen and thymus cells from rats injected with a large dose of antigen (ovalbumin, 100 mg, i.p.) 24 hr in advance produced substantial amounts of IDS in culture within 24 hr in the absence of mitogen or additional antigen, but not the cells from rats injected with an immunizing dose (1 mg) of the same antigen. The cells producing IDS in the spleen were shown to be adherent to glass wool, and those in the thymus were partially so. IDS production by antigen-stimulated spleen cells was abrogated by injecting rats with bromodexyuridine (BUdR) at 0 and 12 hr after the ovalbumin. These findings suggest that a subpopulation ofadherent spleen cells (possibly resembling cortical thymocytes), which begins to proliferate within a few hours after a large dose of systemic antigen, produces IDS. This may account for increased nonspecific suppressor activity observed at the same time.


Are Actinomyces viscosus antigens B cell mitogens? An extracellular heteroglycan (ECHG) and a sonicated cell supernatant (SCS) of Actinomyces viscosus Ny 1 induced strong lymphocyte proliferation. This was shown with spleen and thoracic duct cells form germfree rats and confirmed with cells from conventional "nude" mouse spleens. Spleen cells developed direct plaque-forming cells against densely coupled TNP-SRBC. The mitogenic property of ECHG was diminished considerably after mild alkaline hydrolysis for lymphocytes form rat spleens and was totally abolished for cells from "nude" mouse spleens. These results suggest that ECHG and SCS have B cell mitogenicity.


Excretion of DNA by purified human lymphocyte subpopulations. A large proportion of DNA synthesized in vitro by human lymphocytes stimulated with plant mitogens or specific antigens is selectively excreted from the cells. To determine if DNA excretion differs among various types of lymphocytes, we examined purified human lymphocyte subpopulations for DNA synthesis and excretion in response to stimulation by L-PHA. The relative proportion of newly synthesized DNA that is excreted by unseparated mononuclear cells, macrophage-depleted cells, T, and B lymphocytes is identical despite great differences in the magnitude of their responses. Low levels of both DNA synthesis and excretion by macrophage-depleted cells and B cells can be increased by reconstitution with macrophages and T cells, respectively. These data indicate that DNA exretion is a general property of lymphocytes stimulated to undergo DNA synthesis by plant mitogens.


Microwaves induce an increase in the frequency of complement receptor-bearing lymphoid spleen cells in mice. A single 30-min exposure of mice to 2450 MHz microwaves (12 to 15 mW/g body weight) in an environmentally controlled waveguide facility induced a significant increase in the proportion of complement-receptor positive lymphoid cells in the spleen. This effect was further enhanced by repeated (three times) exposures, which in addition produced a significant increase in the proportion of Ig+ cells. The proportion of theta-positive cells and the total number of spleen cells remained unchanged.


Suppression of natural killer cell activity with radioactive strontium: effector cells are marrow dependent. NK cells with lytic capacity for Moloney leukemia virus-induced lymphomas have previously been found to occur spontaneously in spleens from nonimmune adult mice. Here, 89Sr-treatment is shown to suppress NK cell function in adult mice without similarly affecting other cell-mediated immune reactions. Thus, selective 89Sr-sensitivity distinguishes NK cells from other killer cell types. The present results indicate that in vivo a functional bone marrow is needed for generation and maintenance of NK activity.


A microassay for antibody binding to tumor cell surface antigens using 125I-labelled protein a from Staphylococcus aureus. A sensitive assay for antibodies bound to the surface antigens of adherent tumor cells using protein A from Staphylococcus aureus is described. Cells, as monolayers in microtest wells, are incubated first with immune or control serum, and then with 125I-labelled protein A (IPA), which binds to IgG antibodies bound to cell surface antigens. The IPA assay is shown to be more sensitive than the conventional isotopic antiglobulin (IAG) assay, because IPA binds to a lesser extent to IgG bound non-specifically to cells.


Heparin-induced coagulopathy.

Heparin-induced coagulopathy. Intravenous heparin, at doses of 3.0 U./gm of body weight, produced an intravascular coagulopathy in rats which was manifested by intestinal tract hemorrhage, a reduction in plasma fibrinogen concentration, a rise in fibrinogen-fibrin degradation products, and the absence of a rise in platelet count noted in the control animals. This coagulopathy could not be produced by conventional anticoagulant doses of heparin or the injection of large doses of heparin in the presence of protamine sulfate. Specific studies excluded hypoxemia, metabolic acidosis, and endotoxemia as possible etiologic factors. The coagulation abnormalities observed in this study differ from those produced by injection of other polyanionic substances but their precise pathogenesis is still uncertain.


Thymus-dependent lymphoid regions in the spleen of the lizard, Calotes versicolor. This paper describes the distribution of thymus-derived lymphocytes in the spleen of the lizard, Calotes versicolor. Our studies reveal the existence of red and white pulp in the spleen. The red pulp consists of collagenous fibrous septae containing lymphocytes, erythrocytes and sinuses. The white pulp is an area enclosed by fibrous septae, containing lymphocytes, reticular cells and arterioles. The region around the arteriole in the white pulp was depleted of lymphocytes either one month after adult thymectomy or after anti-thymocyte serum treatment. The repopulation of this region was observed in sham-thymectomized controls, but not in thymectomized lizards. Therefore, this peri-arteriolar region may be designated as a thymus-dependent area, as described previously in higher vertebrates. The significance of these findings is discussed in relation to the phylogeny of the immune system.


Factors influencing the differential sorption of odorant molecules across the olfactory mucosa. By use of a flow dilution olfactometer, tritium-labeled odorants were presented through the external naris to the bullfrog's intact olfactory sac. After stimulation the animal was frozen in liquid nitrogen. The dorsal surface and eminentia of the olfactory sac were then removed and sawed into sections perpendicular to the long axis of the mucosal surface. Each section was dissolved in a tissue solubilizer and counted in a liquid scintillation system. The amount of radioactivity in each section was used to estimate the number of odorant molecules it sorbed. For tritiated butanol there was a significant decrease in radioactivity from the section containing the external naris to that overhanging the internal naris. The steepness of the gradient was unaffected by a rather large range of stimulus flow rates, volumes, and partial pressures. Only when these parameters were pushed to extreme physical limits did this gradient change significantly. When the stimulus was presented through the internal rather than the external naris, the butanol gradient reversed its direction, decreasing from the internal to external. Unlike butanol, tritiated octane presented through the external naris was rather evenly distributed among the mucosal sections. That is, octane showed no distribution gradient across the mucosa. These results complement previous electrophysiological data that suggested a "chromatographic-like" differential sorption of odorant molecules across the mucosa.


Time course and magnitude of effects of changes in tonicity on acetylcholine release at frog neuromuscular junction. 1. The time course for the changes in miniature end-plate potential (min epp) frequency and in epp amplitude produced by alterations in the tonicity of the Ringer at the frog neuromuscular junction was studied. The relations between the tonicity and min epp frequency as well as epp amplitude were also investigated. 2. The change in min epp frequency occurred within 1 min after the start of the change in the tonicity of the extracellular solution. Following a shift to a hypertonic solution, the min epp frequencies were often maintained at a relatively steady, elevated level, even with large (+100 mosM) changes in tonicity. In other instances the elevation was transitory like the reported data for the rat neuromuscular junction. Essentially the same results were obtained in very low Ca2+-Ringer. Unlike the rat neuromuscular junction, the final level after hours of the increased min epp frequency caused by raising the osmolarity by more than 75 mosM was well above the control level. Following the return from a hypertonic to an initial solution there was a prompt decrease in min epp frequency to about the initial level; there was no indication of the transitory depression in min epp frequency following the return from hypertonic solution that has been reported in mammals. 3. Until the osmolarity of the Ringer reached about 420 mosM, the frequency of min epp continued to rise along a line relating log (min epp frequency) to (osmolarity)0.5. When the osmolarity exceeded 460 mosM, the relation started to level off. 4. The hypothesis that the min epp frequency in a Ringer with a given increased tonicity is a fixed multiple of the frequency in normal Ringer is not in accord with the data. 5. The decrease in epp amplitude caused by markedly hypertonic solutions also came about within 1 or 2 min after the start of the change in the tonicity of the solution surrounding the nerve terminal. 6. Hypertonic solutions did not appear to affect facilitation. 7. Below 360 mosM increasing the tonicity of the Ringer had little effect on the amplitude of epp. Above this level the amplitude decreased as the tonicity increased. At a given junction an increase in tonicity in a range above 360 mosM can cause an increase in min epp frequency and a decrease in epp amplitude. 8. The results are discussed in terms of the theories proposed to account for the effects of osmolarity on synaptic function. Two theories--the water flow hypothesis (11) and the barrier of water hypothesis (2)--do not fit with the results. The two other theories--calcium elevation (1) and screening of surface charges (3, 13, 21)--fail to account for important aspects of the results and therfore cannot be accepted without substantial modifications. None of the theories devised to account for the increase in min epp frequency predicts the falloff in frequency and in evoked quantal release that occurs in highly hypertonic solutions.


Chronic cerebellar stimulation and developmental reflexes. The changes in motor function in cerebral palsy patients produced by chronic anterior lobe cerebellar stimulation were documented with the Milani-Comparetti developmental scale, which allows comparison betweenfunctional gains and reflex patterns. Two patients with marked motor retardation and primitive reflexes were followed serially over several months of stimulation. Using these measures, the efficiency of chronic stimulation can be assessed, although many years will be needed to evaluate its role in treating motor disabilities.


Lipoma of the cerebellopontine angle. Case report. The authors present a surgical case of a lipoma of the cerebellopontine angle in a 28-year-old woman.


A nerve stimulator for the cerebellopontine angle. Technical note. A fine-tipped bipolar nerve-stimulator probe has been developed and has proved especially useful in intracranial surgery of the cranial nerves.


Convective diffusional analysis for drug transport through a tubular polymeric membrane. The transport of three p-aminobenzoate esters (ethyl, butyl, and hexyl) through a tubular dimethyl polysiloxane membrane into a flowing liquid was investigated. The tubular configuration permits the exact determination of the convective diffusional contribution to membrane transport with models that account for fluid hydrodynamics. The observed transport behavior ranged from complete convective diffusion control for the hexyl ester to complete membrane control for the ethyl ester; the butyl ester exhibited a change in control with flow rate. The implications of convective diffusional considerations to intestinal absorption and dissolution studies are discussed.


Direct comparison of three isotopic release microtoxicity assays as measures of cell-mediated immunity to Gross virus-induced lymphomas in rats. Three isotopic release microtoxicity assays--[125I]5-iodo-2'-deoxyuridine release assay (IRA), 51Cr release assay (CRA), and [3H]proline release assay (PRA)--have been utilized to measure cell-mediated immunity to (C58NT)D, a Gross virus-induced lymphoma in rats. These studies were designed so that all three assays were done under physical conditions as comparable as possible between the assays. A considerable difference was noted in the ability of one or another target cell to function well in each assay. The tissue culture line of (C58NT)D proved an excellent target cell in the long-term assays, whereas the ascites line was inadequate in these same long-term assays. The monolayer Gross virus-induced tumor cell line ERTh/G was resistant to lysis in the short-term CRA but functioned well in the long-term assays. The autologous and thymus cell controls utilized in these studies were reasonably neutral baseline controls for the evaluation of both normal and immune activity. Although all three assays were capable of measuring both natural and immune activity in this systemthe PRA appeared more sensitive at 24 hours and the IRA at 48 hours, whereas the CRA activity with these target cells was only useful in the short-term assays.


Immunologic function in humans before and after hyperthermia and chemotherapy for disseminated malignancy. Multiple immunologic parameters were studied in three patients prior to and after hyperthermia treatment for disseminated malignancy. Two patients had malignant melanoma and received chemotherapy during the hyperthermia treatment. One had adenocarcinoma of the stomach and received no concomitant chemotherapy. Rapid rosettes as a measure of thymus-derived lymphocytes (T-lymphocytes) were found to increase significantly after therapy (P less than 0.05) both in percentage and absolute numbers. There was no change in the numbers or percentages of other markers for T-lymphocytes or bone marrow-derived B-lymphocytes. Complement profiles revealed a significant decrease in C3 (P less than 0.005) after hyperthermia but no change in levels of other components of the alternate pathway. Antibody-dependent lymphocyte-mediated cytotoxicity and polymorphonuclear cell-mediated antibody-dependent cytotoxicity were also depressed after hyperthermia. No change was observed in immunoglobulin levels with hyperthermia therapy. Results indicated that hyperthermia may favorably alter the immune balance between tumor and host in selected instances.


Isolated aortic valve replacement in patients older than 65 years. The results of 196 isolated aortic valve replacements in patients older than 65 years were analyzed. Eighty-four percent of patients were in New York Heart Association (NYHA) functional class III or IV preoperatively. The operative mortality was 12% for all cases and 9% for elective cases. Actuarial probability of five-year survival was 55% for the entire group and 61% for discharged patients. Myocardial failure and congestive heart failure were the most common causes of early and late postoperative death, respectively. Embolic strokes occurred in 16% of discharged patients and caused substantial disability in 9%. At the termination of the study, 94% of surviving patients were in NYHA class I or II, and none were in class IV. Aortic valve replacement in elderly patients entails reasonable operative risk, and results in satisfactory postoperative rehabilitation.


Mechanisms of occlusion of saphenous vein--coronary artery "jump" grafts. Seven patients with 12 end-to-side and eight side-to-side saphenous vein graft-to-coronary artery anastomoses were studied at autopsy. Postmortem coronary arteriograms and serial histologic sections of the anastomoses were performed. Occlusive changes occurring in the coronary arteries tended to be at the ends of the arteriotomy and could be attributed to reduction of arterial circumference from its eversion and utilization in the anastomotic connection, or from thrombosis along the suture line or from both. The mechanisms of arterial occlusion were the same in side-to-side and end-to-side anastomoses. In addition to the occlusive changes at the anastomosis, "jump" grafts with two anastomoses were susceptible to rolling or kinking of the graft, apparently as a result of torsion from placement of the side-to-side anastomosis.


Occlusive changes at the coronary artery--bypass graft anastomosis. Morphologic study of 95 grafts. Ninety-five bypass graft anastomoses in 52 patients dying up to 4 years after direct coronary revascularization were studied at autopsy by angiograms and serial histologic sectioning of the graft-artery anastomosis. When new coronary occlusions and narrowings occurred, they were adjacent to either the proximal or distal ends of the anastomosis and were due to compression or loss of circumference of the arterial lumen (40 per cent), thrombus formation (40 per cent), mural dissection of the coronary wall (8 per cent) or the combination of compression and thrombosis (12 per cent). Small coronary artery diameter, local atheromas, and extension of the arteriotomy into a branch vessel were significant factors predisposing to occlusive changes. The findings emphasize the importance of careful artery selection for bypass, the need to avoid local vascular disease and branch-points, and the technical difficulties encountered in the presence of local vascular lesions or small coronary arteries.


Left main coronary bypass for aberrant (aortic) intramural left coronary artery. This report describes the first saphenous vein bypass graft from the aorta to the left main coronary artery for an aberrant left main coronary artery arising from the anterior sinus of Valsalva. A 20-year-old college student had a cardiac arrest and documented ventricular fibrillation while jogging. He was resuscitated. An anomalous left main coronary artery, arising anteriorly from the right coronary sinus, was demonstrated at operation to be within the wall of the aorta. Following aorta--left main coronary artery bypass with the saphenous vein, results of a stress test were normal, and cardiac catheterization revealed the left coronary system to be entirely supplied by the graft.


Long-term clinical and hemodynamic studies after ventricular aneurysmectomy and aorta-coronary bypass. Late clinical and hemodynamic evaluations in 18 patients with ventricular aneurysmectomy and aorta-coronary bypass are presented. Tne patients had significant obstructive lesions in two  major vessels (55 per cent), and 6 had extensive three vessel disease (33 per cent). In 13 patients, 21 aorta-coronary saphenous bypass grafts were performed in addition to aneurysmectomy. The operative mortality rate was 11 per cent. One patient died suddenly 5 months after the operation (one year mortality rate 17 per cent). The 15 surviving patients have been followed up for 12 to 41 months (average 24 months). Clinical results were considered excellent in 2 patients who have been asymptomatic (Class I, N.Y.H.A.). Nine others were considered to have good clinical results (Class II). Five patients have continued to have congestive heart failure and angina on minimal effort (Class III or IV). Six of the 11 patients considered to have excellent or good results underwent postoperative hemodynamic studies 6 to 34 months after the operation. A significant increase in cardiac index was documented in all 6 patients. Paradoxic movement was not detected in any of the postoperative ventriculograms. Five of the seven venous grafts inserted were patent. Elevated left ventricular end-diastolic pressure (LVEDP), low cardiac index, and a persistent dyskinetic area in the left ventricle were found in 2 patients considered to have poor clinical results. Clinical and hemodynamic evaluations have shown a significant improvement in most patients surviving ventricular aneurysmectomy. However, postoperative systemic embolism, myocardial infarction, progression of coronary artery disease, transient cerebral ischemic attacks, graft occlusion, arrhythmias, and mitral regurgitation in previously prolapsed mitral valve leaflets account  for progressive disability and limited activity after a successful operation.


Measurements of autonomic function before, during, and after transcutaneous stimulation in patients with chronic pain and in control subjects. Transcutaneous stimulation (TCS) has been reported to modify peripheral blood flow, skin temperature, blood pressure, and heart rate, all of which are under the influence of the autonomic nervous system (ANS). In 20 patients with intractable pain and in 10 control subjects, TCS was not found to alter significantly any of the observed properties of ANS function of control subjects or patients, except to decrease skin impedance in the patient group after the stimulation. Patients reporting pain relief during TCS were found to have significantly higher systolic blood pressure under all test conditions than patients reporting no pain relief (p less than 0.05). No other significant differences were found between these two groups. Finally, in the patients, no significant localized autonomic changes were demonstrated in the painful area by comparison with the homologous body part.


Neonatal hepatitis and alpha-1-antitrypsin deficiency. The prognosis in five patients. Five patients with alpha-1-antitrypsin deficiency (PiZ) are reported. All these patients presented with the neonatal hepatitis syndrome and two fo them had developed cirrhosis at ages 5 and 8 years, respectively. Three patients, ages 1, 9 and 21 years, are asymptomatic. The oldest patient, 21 years of age, has only mild histologic changes in the liver. The prognosis for patients with alpha-1 antitrypsin deficiency (PiZ) presenting with neonatal hepatitis is not necessarily grave, a finding that differs from previous observations.


Acoustic neuroma causing subarachnoid haemorrhage. The case reported in this paper appears to be the first example of spontaneous subarachnoid haemorrhage complicating an acoustic tumour. The only comparable case is one of a pontocerebellar angle tumour, but its histological nature is unstated. The excellent response to surgical management has been gratifying, and the patient has remained well over an extended period of surveillance.


High-dose methotrexate therapy.

High-dose methotrexate therapy. Eighty-six adults with malignant disease were given high doses of methotrexate with folinic acid rescue, with acceptable toxicity. Protocal violations in two cases led to death. The results of therapy in some diagnostic groups are encouraging.


Von Willebrand's disease: combined qualitative and quantitative abnormalities. Studies of von Willebrand's disease typically show either a quantitative or a qualitative abnormality of the factor VIII/von Willebrand factor protein. In studies of five patients we found a combination of quantitative and qualitative defects. Quantitative abnormalities included decreased levels of procoagulant, antigen and von Willebrand factor activities in the plasma and in the column fractions of cryopercipitate from gel chromatography. Qualitative abnormalities included annormal migration or shape of the crossed antigen-antibody arcs, disproportionate reduction of von Willebrand factor in relation to antigen, altered gel elution patterns of procoagulant, and von Willebrand factor activities the negative carbohydrate stain of the VIII protein on polyacryl-amide-gel electrophoresis and, in two patients, a decreased sialic acid content of factor VIII protein. Our studies indicate that von Willebrand's disease comprises both quantitative and qualitative defects.


[The stress ulcer in the neurosurgical disease and the warning data (author's transl)]. Twelve cases of G.I. tract bleeding, treated in the Department of Neurosurgery, Kobe University School of Medicine, were analyzed with the special reference to clinical data before the episode of G.I. tract bleeding. These 12 cases consist of 10 cases of aneurysm, 1 of metastatic brain tumor, and 1 of chronic subdural hematoma. The anterior communicating artery has the majority in the case of aneurysm by 50 per cent. In each case, C.B.C., bleeding time, coagulation time, serum electrolytes, serum glucose, glucosuria, steroid administration, blood pressure, hyperthermia, and water balance were reviewed retrospectively. Only serum glucose showed the specific abnormality with the average of 135 mg/dl (Hoffman method) before the episode of G.I. tract bleeding among them. In 1931, Cushing reported the clinical case of G.I. tract bleeding with intracranial disease and also experimental G.I. tract bleeding, in which discussion he concluded only parasympathetic nerve system has the involvement in the mechanism. In 1952, French, however, certified also the involvement of sympathetic nerve system in G.I. tract bleeding with the experimental results. In addition to this report, many experimental works under the standpoint of hypothalamus involvement in stress ulcer were undertaken. On the other hand, hyperglycemia and glycosuria in the stress have been reported as the results of clinical and experimental works. In 1971, Frohman concluded that only ventromedial nuclei has relation to the control of glycogen metabolism in hypothalamus from his experimental results. We concluded that this hyperglycemia before the evidence of G.I. tract bleeding due to the involvements of hypothalamus or other parts of C.N.S. in the neurosurgical disease is one clinical sign of the stimualtion to sympathetic nerve system that also can be a cause of G.I. tract bleeding, and this hyperglycemia before the evidence of G.I. tract bleeding should be paid attention as a warning datum.


Extra-axial growth of fourth ventricle tumors - angiographic changes. Tumors of the fourth ventricle, particularly ependymomas, may grow out of the ventricular cavity into the cerebellopontine angle cistern and the subarachnoid cisterns. The angiographic changes will then suggest both the intra- and extraventricular location of the tumors. The ependymoma is the most prone of the fourth ventricle tumors to demonstrate this pattern of growth.


Angiographic findings with an intracranial gumma. The case of a syphilitic gumma in the cerebellopontine angle is reported. The angiographic appearance of neurosyphilis is similar to that of cerebral inflammatory processes of all varieties. The hypervascularity of the blush shown on angiography is due to hyperemia surrounding the focal necrosis. Vasoconstriction and dilatation may be seen with meningovascular syphilis and gummata. This case showed a focal hypervascularized lesion unlike the avascular lesions described in the textbooks.


Propagation of Pneumocystis carinii in vitro. Pneumocystis carinii was propagated in vitro with primary embryonic chick epithelial lung (CEL) cells. Viability and growth of the organism were demonstrated by direct observation of the reproductive cycle in the Sykes-Moore chamber, serial passage with an increase in the number of mature cysts forms, the cytopathic effect of the organism on cell culture, and inhibition of growth of the organism by specific antiserum and pentamidine isethionate. Attempts to culture P. carinii indefinitely were not successful. However, cyst forms derived from murine and human sources increased 100-fold and 10-fold, respectively, during CEL cell culture passages. Serial passage of trophozoites alone resulted in the development of typical CPE and a maximum number of 2.8 X 10(3) cyst forms. Autoradiographic methods demonstrated active DNA, RNA, and protein synthesis within the cyst and suggest that metabolic interaction between the host cells and the organisms occurred. The nature of the attachment of P. carinii to the host CEL cell was clearly discernible by scanning electron microscopy (SEM). In the reproductive cycle a vegetative cell (designated "trophozoite") attached by tubular expansions to the hhed without entering the cell. Sporozoites developed within the detached young cyst, reaching a maximum number of eight within the mature cyst. Excystment occurred through single or multiple sites in the cyst wall, after which the released trophozoite attached to a new host cell.


[Major esophageal hemorrhages of unusual origin. A poorly known entity]. Nine cases of severe gastrointestinal haemorrhage as the presentation of oesophageal lesions are reported. Haemorrhages complicating oesophageal varices, carcinoma of the oesophagus and oesophageal surgery were eliminated from the study, as were haemorrhages associated with hiatal hernias or Mallory-Weiss syndrome. Amongst these 9 cases of severe haemorrhage, 5 were due to an ulcer of the oesophagus, 2 to acute oesophagitis, 1 to rupture of the oesophagus and one to a traumatic aneurysm of the aorta rupturing into the oesophagus. The location and cause of bleeding were determined only 3 times before operation and only when a fibroscopic examination had been carried out. At fibroscopy, an irregular lesion responsible for a severe haemorrhage has every chance of being a benign ulcer and not a carcinoma. Treatment directed at the aetiology was carried out in 6 cases and success obtained in 3 patients. The aetiological diagnosis who was not made in 3 patients, who died. The aetiology, diagnosis and therapeutic approach required in such cases of severe haemorrhage of oesophageal origin are considered. In the light of these cases, a possible oesophageal aetiology should be considered as a possible cause of an unexplained severe gastrointestinal haemorrhage. An attempt at proving such a hypothesis by fibroscopy is necessary and, in general, urgent thoracotomy since the bleeding is in most instances of arterial origin.


Macromolecular factor VIII complex: functional and structural heterogeneity observed in von Willebrand swine with transfusion. The physiologic activities concerned with hemostasis and associated with the Factor VIII macromolecular complex were investigated in swine with von Willebrand's disease after infusion of cryoprecipitate, a lyophilized Factor VIII concentrate, or porcine serum. Immediately after each infusion the various activities antihemophilic factor, von Willebrand platelet aggregating factor, and Factor VIII-related antigen, were elevated in approximate proportion to dose and the bleeding time was shortened.There was a late secondary rise in antihemophilic factor. During the period after infusion, there was a differential fall-off of the various activities, with the bleeding time effect lost first, followed by the von Willebrand platelet aggregating factor and then by the Factor VIII-related antigen. The plasma from swine with von Willebrand's disease late after infusion contained high levels of antihemophilic factor without other detectable activities of the complex. Antihemophilic factor, free of the other components, obtained from plasma from swine with von Willebrand's disease either before or late after infusion eluted from agarose gel columns both as high and lower molecular weight material, unlike normal antihemophilic factor, which had a high molecular weight. In contrast, on ultracentrifugation the antihemophilic factor in these plasma sedimented slowly, even though chromatographically the plasmas contained both high and low molecular weight factor. All of the Factor VIII complex activities in normal porcine plasma sedimented rapidly. These studies demonstrate the heterogeneity of  the Factor VIII complex and the apparent dependence of its chromatographic and sedimentation behavior on the functional activities associated with the complex.


Partial chemical characterization of Ia antigens derived from murine thymocytes. Previous chemical studies attempting to demonstrate Ia antigens on mouse thymocytes have given contradictory results. We attempted to resolve the question of whether Ia antigens exist on thymocytes (defined as thymus cells that bear a T cell marker) by isolating strain C3H thymocytes free of other contaminating cells using the fluorescence-activated cell sorter, and then chemically testing the purified populations for Ia antigens. Immunoglobulin-negative thymus cells and thymus cells selected with a rabbit antiserum to mouse brain were the two populations of thymocytes labeled with [3H]leucine after sorting. Radiolabeled proteins were solubilized with the non-ionic detergent Nonidet P-40, reacted with anti-Ia antiserum, and analyzed by electrophoresis on discontinuous sodium dodecyl sulfate/polyacrylamide gels. Ia antigens were recovered from both cell populations. These antigens were synthesized by thymocytes and were found on molecules composed of two chains of molecular weight 33,000 and 25,000, respectively, similar to Ia antigens derived from spleen cells. Assuming that all thymocytes bear similar amounts of Ia antigens, we estimated that they have approximately 1/50 the amount of Ia antigens that spleen cells do.


Heterogeneity of the cytotoxic response of thymus-derived lymphocytes after immunization with influenza viruses. Immunization of mice with serologically distinct type A influenza viruses results in development of highly crossreactive populations of cytotoxic thymus-derived lymphocytes (T cells). This can be shown both at the level of effector function and of priming for an enhanced response after challenge with another type A virus. Cytotoxic activity is demonstrable, in both the primary and secondary situations, only for H-2 compatible interactions. Further analysis by competitive inhibition experiments indicates that some of the T cell clones generated are specific for the virus used to immunize, while others are much less restricted. Secondary stimulation may result in preferential stimulation of the crossreactive T cells if the type A viruses used are very different serologically. When more closely related viruses are used, however, some degree of specificity is seen for the challenge virus. Even so, the patterns of crossreactivity observed are complex, and cannot be readily predicted on the basis of known serological relationships between surface hemagglutinin and neuraminidase antigens of type A influenza viruses.


[Simple and rapid heterologous method of radioimmunochemical determination of luteinizing hormone in the blood serum of rats. 1. Basic principles of the method]. Possibilities of creation of a radioimmunochemical method (RICM) of determination of the luteinising hormone (LH) were studied. The method is based on the application in the binding system of a water-insoluble immunosorbent obtained by copolymerization of antisera with isobutyl ester of chlorocarbonic acid. The antisera were obtained by immunization of rabbits with a highly-purified LH isolated from sheep hypophyses. The method is rapid and precise, and is carried out in a single stage. Significant results of RICM in the LH determination can be obtained with the minimal two-hour incubation time. Using different amounts of the immunosorbent in the reaction mixture with the same indicator dose of I125-LH it is possible to vary the method sensitivity (from 0.05 to 5 ng/test).


Cytolytic thymus-derived lymphocytes specific for allogeneic stimulator cells crossreact with chemically modified syngeneic cells. Mouse spleen cells cocultured with irradiated allogeneic stimulator cells develop cytolytic effector cells capable of lysing 51Cr-labeled syngeneic trinitrophenyl-derivatized tumor or spleen targets and to a lesser degree unconjugated tumor cells in addition to the allogeneic stimulator cells. Lysis of trinitrophenyl-syngeneic targets was inhibited competitively by cold trinitrophenyl-syngeneic tumor or spleen targets as well as by cells bearing the allogeneic stimulator H-2 haplotype demonstrating the immunological specificity of the interaction. Allogeneic H-2 specificities may, therefore, be considered variants of modified autologous H-2 specificities against which cytolytic thymus-derived clones potentially exist that are capable of exerting immunological surveillance.


Biochemical catalysis involving coenzyme B-12: a rational stepwise mechanistic interpretation of vicinal interchange rearrangements. A mechanism is proposed for the catalytic action of coenzyme B-12 which is consistent with current knowledge of organometallic reactions and with the experimental data now available from biochemical studies. A key feature of the proposal is an electrocyclic cleavage of the coenzyme that reduces cobalt and also leads to a 1,19-seco-corrin. The seco-corrin serves as a tridentate ligand about Co(I). This arrangement permits the metal to take part in the kinds of organometallic reactions that are ideal for coenzyme B-12 catalysis, including oxidative addition and its reverse, reductive elimination. It is further proposed that the rearrangement steps involve cobaltcarbene complexes.


The keloidal diathesis, a resistant state to malignancies? We report clinical observations and immunological and laboratory studies whcih suggest that the tendency toward skin malignancies and the keloid diathesis may be two opposing conditions. We conclude that it is quite possible that a keloidal person, armed with a hypersensitive cell-mediated immune response condition and an overactive enzyme system, has a safeguard against acquiring skin malignancies.


Computed tomographic diagnosis of suprasellar masses by intrathecal enhancement. Ten suspected suprasellar mass lesions were evaluated by CT cisternography (CTC). In each case the cisterns could be defined. Suprasellar mass lesions were found in six. Precise assessment of the anterior, lateral, posterior, and superior extent, made by metrizamide CTC, was verified at surgery. In two masses the intravenously enhanced scan was not diagnostic, while a lesion was visualized by intrathecal (metrizamide) CT. The major diagnostic entity was an enlarged third ventricle. When a chiasmic lesion is suspected and the conventional scan is negative, metrizamide CTC is the examination of choice.


Radionuclide skeletal survey for pediatric neoplasms. A prospective study comparing the radionuclide skeletal survey with the radiographic survey was performed for 159 children with known primary neoplasm. Forty-four showed radionuclide evidence of metastases, while only 14 had radiographic evidence. Thus, 68% of metastases were discovered by bone imaging alone. No false negative images for metastases were obtained, though there was one false negative image for primary tumor. Radionuclide skeletal survey is recommended as the primary evaluation for bony metastases in all children with neoplasms. Suspect areas should then receive complete radiographic examination.


Influence of freezing and storage methods on concentrations of brain DOPA and 5 HTP after pretreatment with a decarboxylase inhibitor. The concentrations of DOPA and 5 hydroxytryptophan in rat brain were decreased by freezing on dry ice rather than in liquid nitrogen, and by a 5 minute delay in removing and freezing the brain by either method. The effect of post-mortem delay in freezing was greater for 5 HTP than for DOPA.


[The biochemistry of collagen and the locomotor apparatus. Hereditary diseases of connective tissue and rheumatologic diseases (part 2)]. In lathyrism, the toxic agent directly blocks the development of the transverse links in the collagen fibre, while in bovine dermatosparaxis there is a deficiency of procollagen peptidase. Of the many clinical forms of the Ehlers-Danlos syndrome, some are due to a deficiency of lysine oxidase, others to deficiency of lysine hydroxylase and still others to procollagen peptidase deficiency. The essential deficiency in Marfan's disease is still unknown, but that in homocystinuria causes blocking of the groups necessary to form the transverse links. In osteogenesis imperfecta, which is probably a heterogeneous group, there is deficient consolidation of the collagen fibrils, the cause of which is still unknown. One possibility is a quantitative imbalance between chains of different types. In scleroderma there is excessive synthesis of an apparently normal connective tissue; the cause of this is also still unknown.


Distribution of B, T, and O lymphocytes in blood and tissues of normal humans reflecting a kinetic model. The occurrence of the lymphocyte subpopulations TE, BFc, BC3, BFc + C3, BIg, and O-, percentage as well as absolute, was studied in lymphocyte suspensions from tonsils, abdominal lymph nodes, spleens, bone-marrow aspirates, and at the same time in venous blood. The absolute lymphocyte content (number of lymphocytes per g tissue) was highest in the abdominal lymph nodes, lower in the spleens and tonsils, and lowest in the blood. TE lymphocytes were found in the significantly highest percentage, 60%, in the blood. B lymphocytes, comprising BFc + C3 plus BIg, were present in the highest percentage in the bone marrow: 74%. Tonsils, spleens, and abdominal lymph nodes contained fewer B lymphocytes, and the blood fewest: 39%. A significant correlation was found only between the absolute numbers of T and B lymphocytes. A relationship between the absolute number of T lymphocytes and the total number of B lymphocytes as well as fractions thereof was thus demonstrated in the various tissues and in the blood and also between the blood and the tissue. O- lymphocytes were found in bone marrow, lymph nodes, and spleens, apparently as markerless precursors of other subpopulations. The main conclusion of the study is: In the lymphocytokinetic system the T lymphocytes must play a guiding role as an afferent vector, trigging the B fractions which thus constitute the efferent vector of the system.


T-cell regulation of immunoglobulin synthesis and proliferation in pokeweed (Pa-1)-stimulated human lymphocyte cultures. In pokeweed (Pa-1)-stimulated human lymphocyte cultures, T cells are essential for the survival, proliferation, plasma-cell development, and high-rate Ig secretion of B cells. Their effects are T-cell-specific in the sense that B-cell stimulation does not take place in the absence of T cells even when fibroblasts or monocytes are provided. The experimental system is the most effective model for activation of human B lymphocytes so far described. Plasmablast development requires approximately 7 days in culture. In T + B-cell cultures established at 1 X 10(6)/ml (1 X 10(4)/mm2) initial cell density, plasma cells can secrete, on the average, as much as 40-70 pg IgM or IgG per cell per day. When the initial T-cell density is increased well above 1.0 X 10(6)/ml, a T-cell-mediated depression of Ig synthesis becomes predominant. Thus, in the pokeweed model T-cell effects represent a balance of helper and suppressor influences. The study also shows that B cells are heterogeneous. A non-adherent IgG-committed (smIg-?) TONSIL B-cell population seems to be less susceptible to T suppressor effects than normal tonsil B cells. This subset proliferates particularly well and synthesizes large quantities of IgG in the presence of large initial proportions of T cells.


Tentative identification of arginine vasotocin in the bovine pineal extract prepared by in the bovine pineal extract prepared by the Milcu-Nanu method. The bovine pineal extract prepared by the Milcu-Nanu method (BPE-MN) has frog bladder, rat antidiuretic and rat uterine activities susceptible to tryptic digestion. The ratios of these activities are characteristic of arginine vasotocin (AVT). Desalting on Amberlite CG25 and partial purification of BPE-MN by free-flow continuous electrophoresis (FFCE) allow the separation of some fractions with AVT-like biological activities and with a carrier-free electrophoretic mobility similar to the synthetic AVT. The results strongly suggest the presence of AVT in the bovine pineal extract prepared by the Milcu-Nanu method.


Size distribution of in-vitro-induced antigen-specific helper and suppressor cells. Specific helper and suppressor cells were induced in vitro with low and high doses, respectively, of protein antigen in Marbrook cultures. On different days of culture the cells from helper and suppressor cultures were size-fractionated by 1-g velocity sedimentation and the different fractions tested for specific activity in secondary co-operative cultures after challenge with protein-hapten conjugate. We observed a change in the size distribution of both helper and suppressor cells during their in vitro differentiation; the first helper and suppressor cell to be detected is a large (blast) cell, whereas later both help and suppression is mediated by medium- and small-sized lymphocytes. A difference between helper and suppressor cells with regard to their relative dependence on DNA synthesis and proliferation during their induction was also seen. Although both DNA synthesis and divisions are needed for the optimal generation of both types of cells, some suppressor activity was still induced in cultures treated with mitosis-arresting concentrations of demecolcine (Colcemid). Treatment of helper cell cultures with Colcemid or with 5-bromodeoxyuridine plus light completely abolished the help.


Regulation of in vitro immunocyte activation: origin of and targets for cell-released inhibitors. Supernatants from incubated normal mouse spleen cells suppressed the DNA synthetic response induced by polyclonal B- and T-cell activators and the primary immune response to sheep erythrocytes in normal spleen cells in vitro. The inhibitory effects of the supernatants were found to be dependent on the culture system used. The main cell population producing or releasing the inhibitory factors was nonadherent spleen cells, but macrophages and bone marrow cells could also give supernatants with inhibitory effects. Thymocytes did not release any inhibitors under the same conditions. Separation of the supernatants showed that there were at least two factors with inhibitory effects on proliferation in lymphocytes. One of these factors had a molecular weight of less than 10,000 (not degradable by protease) and suppressed activation by all tested polyclonal B- and T-cell activators. Another factor (a protein with a molecular weight of more than 30,000) inhibited activation by some polyclonal B-cell activators (lipopolysaccharide, type III pneumococcal polysaccharide) whereas the proliferation induced by dextran sulfate was less or not at all affected under the same culture conditions. Trivial mechanisms for inhibition by supernatant factors, such as medium depletion, general toxicity, and accumulation of free thymidine in the medium, were excluded.


Inhibition of spontaneous and antibody-dependent cellular cytotoxicity by sera and isolated antiglobulin preparations from rheumatoid arthritis patients. Using sera from patients with rheumatoid arthritis and antiglobulin preparations obtained from such sera, inhibition of human lymphocyte cytotoxicity in antibody-dependent (ADCC) and spontaneous cell-mediated cytotoxicity (SCMC) reactions against an allogeneic melanoma cell line (IGR3) has been demonstrated. Fractionation of effector cell preparations indicated that Fc-receptor-bearing lymphocytes were operative in both reactions. Removal of phagocytic and adherent cells from the effector cell population resulted in more pronounced inhibition of SCMC and ADCC reactions by rheumatoid sera. The results indicate that antiglobulin preparations from human sera containing rheumatoid factor activity can effectively block the cytotoxic activity of Fc-receptor-bearing effector lymphocytes (K cells) in vitro. On analogy with the inhibition of ADCC and SCMC reactions by immune complexes and aggregated IgG, antiglobulin complexes present in the antiglobulin preparations are responsible for this effect.


Mechanism of action of suppressor cells. In vivo concanavalin-A-activated suppressor cells do not directly affect B cells. The addition of a small proportion (10%) of in vivo concanavalin-A (Con-A)-activated spleen cells to normal spleen cell cultures suppressed the primary immune response to sheep erythrocytes (SRBC) but had no effect on the thymus-independent primary immune response to 3,5-dinitro-4-hydroxy-phenacetyl-conjugated lipopolysaccharide. When Con-A-activated cells were added after 24 h, there was no suppression of the anti-SRBC response but rather an enhanced response when few cells were admixed. Con-A-activated cells did not influence activation of normal cells by polyclonal T- and B-cell activators. It is concluded that Con-A-induced suppressor cells do not act on B cells but rather on helper cells (T cells or macrophages) at a very early stage of the immune response to thymus-dependent antigens.


[Transplantation antigens: the individual in biology and psychology]. The persistent defense of biological individuality is explained on the basis of the concept that "foreign" and "self" are complementary phenomena. Since it is the T-lymphocyte of the immune system which recognizes "foreign", its receptor must represent "self". How is this accomplished in immunogenetic terms? An intriguing possibility would be the somatic reduplication and amplification of "self" through random combinatorial principles by means of exclusion: all receptors are formed, but those directed against "self" suppressed while the others (including those identical with "self") remain at disposition. In this way, HLA gene products, rather than representing immune receptors themselves, would prime the formation of the immunological repertoire indirectly. The concept that "self" plays the role of a template copied by the actual recognition system is supported by elementary information theory. Analogies to other, higher biological information systems such as the brain are drawn. Moreover, since neuroscience and psychology are in fact inseparable, the analogies reach even much further. A common blueprint can be traced from primitive cell to cell interactions through molecular immunology to neurochemistry, psychology and philosophy. Particularly Jung's concept of psychological individuation as the never-ending struggle of the human individual for consciousness would precisely fit the role of "molecular individuation" as a means of acquiring the immunological repertoire. In psychological terms "foreign" corresponds not only to the outer world (antigens( but also to our own unconscious (antiidiotypic set) resulting in a similar network of mutual interactions between conscious and unconscious much as between idiotypes and anti-idiotypes.


Photoreceptor outer segments: accelerated membrane renewal in rods after exposure to light. The rate of rod outer segment renewal in Rana pipiens tadpoles under constant light and under diurnal conditions of 12 or 2 hours light per day is significantly increased compared to that in animals in darkness. Furthermore, during 24 hours in light after 6 days in darkness the rate of renewal is three to four times that in darkness. In Xenopus laevis tadpoles the rate of renewal is more than five times greater during the first 8 hours of a normal diurnal cycle than during the following 16 hours. These observations demonstrate that bursts of renewal activity occur as a response to light, and suggest that a normal pattern of light alternating with darness plays a fundamental role in the regulation of rod outer segment turnover.


Infratentorial retrocerebellar cysts.

Infratentorial retrocerebellar cysts. Clinical analyses of ten cases of infratentorial retrocerebellar cysts were performed in order to define specific signs or symptoms and thus improve the preoperative diagnosis. There were not symptoms or signs specific for the cysts. When a tumor in the posterior fossa is suspected by ventriculography, vertebral angiography or pneumoencephalography should be performed to exclude the possibility of a cyst, particularly in patients under 20 years of age. Care should also be taken to look for supratentorial cysts, since multiple cysts are found occassionally. It is necessary to evaluate the therapeutic program for each case, depending on the relationship between the cyst and the subarachnoid space and ventricular system. Postoperative follow-up is important, especially in cases in which only excision of the cyst is performed.


Ligation procedures in the management of portal hypertension. In the 10 year period from 1964 through 1973, 25 patients bleeding from esophageal varices underwent ligation procedures coupled with splenectomy rather than a shunt. These procedures included the transesophageal and transgastric approaches and extragastric ligations. For patients with normal liver function, the risk of this urgent or emergency surgery is comparatively low (two of 12 patients died). The chance of recurrent hemorrhage is high (nine of nine surviving patients), as is the need for subsequent surgery (eight of nine patients). Nevertheless, despite these drawbacks, nine of these 12 patients (75 percent) are alive, and seven have survived 5 or more years. In patients with cirrhosis, the initial operative mortality rate (three of 12 patients) and the subsequent mortality rate (five additional patients) reflect the greater risk because of liver disease. Only five of 13 patients (38 percent) survive, three of whom are alive 5 or more years. In patients with cirrhosis, the initial operative mortality rate (three of 12 patients) and the subsequent mortality rate (five additional patients) reflect the greater risk because of liver disease. Only five of 13 patients (38 percent) survive, three of whom are alive 5 or more years after their initial surgery. These results indicate that there are situations when ligation procedures are valuable, especially in the noncirrhotic patient.


Paraprosthetic-enteric fistula.

Paraprosthetic-enteric fistula. A case of paraprosthetic-enteric fistula is presented and the total reported literature of 21 cases is reviewed. Paraprosthetic-enteric fistula is a complication of aortic revascularization with synthetic prostheses. The entity is characterized by erosion of the gastrointestinal tract by an underlying prosthesis but absence of a true fistulous communication with the aortic lumen. It is both a distinct pathologic entity and a step in the formation of a true aortoenteric fistula with suture line involvement. The most frequent clinical manifestations are sepsis and gastrointestinal bleeding, but nonspecific abdominal pain is present occasionally as well. The distal duodenum is the portion of the gastrointestinal tract involved most commonly. Diagnostic evaluation should include endoscopy, aortography, and barium contrast studies. Venous and femoral arterial blood cultures also should be done in patients presenting with sepsis. Treatment should consist of either graft excision with extra-anatomic revascularization or graft excision alone when dealing with a previously thrombosed prosthesis.


Weak anti-IgA antibodies with limited specificity and nonhemolytic transfusion reactions. Anti-IgA antibodies were studied in sera from patients with nonhemolytic transfusion reactions for which no serological reason had been found. Of the 158 sera that were studied by passive hemagglutination assay using twelve IgA myeloma proteins, 4 samples had class-specific antibodies and 10 samples antibodies with limited specificity. Titers were 1:8 or less. 100 multitransfused hemophilia patients were studied with two IgA myeloma proteins. Four of the sera had anti-IgA antibodies. Normal blood donor sera reacted with IgA only when antigens were first either digested with pepsin or stored for several months as dilute solutions in refrigerator. The results emphasize the need for fresh IgA proteins of good quality when human anti-IgA antibodies are investigated.


[Life-threatening generalized varioloid zoster with zoster pneumonia due to combined immunologic deficiency. Increased contagiousness of this disease]. There is a clear-cut correlation between the clinical course of an herpes zoster and the quality of immune response. A varioloid generalized herpes zoster accompanied by zoster-pneumonia developed in a forty years old female patient with metastasizing breast cancer treated with x-ray and cytostatic drugs. The particularly serious course of this herpes zoster was brought about by combined immune deficiency, consisting of low level of circulating T-lymphocytes (5/mm3) and for a long time lacking rise of complement fixing antibody titer. Under these conditions an enhanced replication and massive haematogenous spread of varicella-zoster virus took place. The increased contagiosity of this case is indicated by a varicella infection in the patient's two children, appearing after a regular incubation period. Patients with cellular or combined immune deficiency are potentially exposed to danger of herpes zoster and may serve as a source of infection for susceptible persons.


New aspects of the life cycle of Pneumocystis carinii. Pneumocystis is recognized as a cause of pneumonitis in vertebrates, including man. So far an extra-cellular life cycle has been proposed; thick-walled cysts are the site of the formation of so-called intracystic bodies. These intracystic bodies are set free and are supposed to develop into free living thin-walled Pneumocysts ("trophic" stage) (Seifert and Pliess, 1960; Vavra and Kucera, 1970; Campbell, 1972). In this paper we present, based on elaborative ultrastructural studies, data on the existence of both intra-cellular and extra-cellular stages of the parasite which result in a new proposal for the life cycle of the parasite. This short communication will be followed by a more detailed description of our observations.


[Histiocytosis X (eosinophilic granulomatosis) of the lung (author's transl)]. Report of a 21-year old male with biopsy-proved (primary) isolated Histiocytosis X (eosinophilic granuloma) localized in the lungs. The disease prefers young males and leads to a non-characteristical fibrosis of the lung in cases of chronical course. Lung biopsy is the only certain method for diagnosis. Morphology, differential-diagnosis and aetiology of this rare disease are also considered.