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PubMed1977

Radioimmunoassay for the antigenic determinants of cholera toxin and its components.

A radioimmunoassay procedure is described for the detection of cholera toxin and its component polypeptide chains. Cholera toxin, A subunit, B subunit, alpha chain, and gamma chain were iodinated by the chloramine T procedure. Radiolabeling did not significantly alter the polyacrylamide electrophoretic migration patterns of the toxin or its components. Moreover, radiolabeled toxin, B subunit, and alpha chain preparations retained substantial ability to bind to intestinal mucosal homogenates. The minimal amount of antitoxin detectable with radiolabeled toxin was 0.04 antitoxin units/ml. Substitution of radiolabeled B subunit, A subunit, and alpha chain for radiolabeled toxin decreased the sensitivity of the test. Radiolabeled gamma chain did not bind to the antitoxin preparation. Competitive inhibition studies, with titrated anti-choleragen serum and radiolabeled toxin or components, indicated that the minimum concentration of toxin detectable was 7.0 x 10(-8) mumol/ml at a 90% inhibition level. The A subunit and alpha chain preparations inhibited the binding of the radiolabeled B subunit to antitoxin sites. Conversely, B subunit inhibited the binding of radiolabeled A subunit and alpha chain to antitoxin. The gamma chain did not show any reaction with antitoxin or cross-reaction with either whole toxin or its components. These results strongly suggest that the A subunit and the alpha chain contain antigenic determinant(s) that are common to the B subunit. The B subunit (beta chain) and the alpha chain of cholera toxin may therefore contain region(s) of chemical similarity.

Antigens, Bacterial↗
PubMed1977

Rabbit intestinal fluid stimulation by an enterotoxigenic factor of Staphylococcus aureus.

An exoprotein of Staphylococcus aureus 100 that elicited a positive ileal loop response in the rabbit model was investigated in this study. The protein, as it occurred in the culture supernatant fluid, could be detected initially in the late log phase of growth under aerobic and anaerobic conditions. It was stable under acidic conditions to pH 2.0 after 24 h at 4 degrees C. Thirty-minute treatments at 80 degrees C destroyed the ileal loop activity whereas similar trials at 70 degrees C had no effect. Although preparations of staphylococcal enterotoxins purified by other investigators did not produce positive ileal loops, the enterotoxigenic activity of the S. aureus 100 supernatant fluid could be neutralized by antisera prepared against enterotoxins A and B. Throughout purification studies, the active moiety reacted serologically with antiserum A. Polyacrylamide gel electrophoresis of the partially purified protein produced migration patterns nearly identical to those of enterotoxin A.

Animals↗
PubMed1978

Different behaviour of 19S thyroglobulin extracted from normal thyroid, simple goitre and toxic goitre in polyacrylamide slab gel isoelectric focusing.

19S thyroglobulin (19S TG) was isolated from normal thyroids, simple goitres and one case of toxic goitre (Graves' disease). With isoelectric focusing on polyacrylamide gel slabs (pH 4--6) several bands for each specimen were detected. The migration patterns of the toxic goitre 19S TG were clearly different from the patterns of normal 19S TG and simple goitre 19S TG.

Electrophoresis, Polyacrylamide Gel↗
PubMed1978

[Molecular weight of lipovitellins during egg development in the crustacean amphipod Orchestia gammarella (Pallas)].

Egg lipovitellins of Orchestia gammarella tested by electrophoresis on gels of different acrylamide concentrations, following the procedure of Hedrick and Smith (1968), display a migration pattern identical to that of proteins with molecular weights of congruent to 3.2 x 10(5) (lipovitellin I) and congruent to 5.5 x 10(5) (lipovitellin II) respectively. Since their molecular weight remains constant during embryogenesis, the changes of their relative mobility in disc gels indicates alterations in their ionic charges.

Animals↗
PubMed1977

[Determination of the molecular weight of vitellogenin and of lipovitellins of Orchestia gammarella, Crustacea, Amphipoda].

Vitellogenin and lipovitellins of Orchestia gammarella, tested by electrophoresis on gels of different acrylamide concentrations, following the procedure of Hedrick and Smith (1968), displays a migration pattern identical to that of proteins of respectively congruent to 4 x 10(5) (vitellogenin), congruent to 3,5 x 10(5) (lipovitellins I and I') and congruent to 5 x 10(5) (lipovitellin II) molecular weights.

Animals↗
PubMed1975

Maturation of B lymphocytes in the rat. II. Subpopulations of virgin B lymphocytes in the spleen and thoracic duct lymph.

Thoracic duct and spleen cells of normal (unimmunized) adult rats were fractionated according to size by 1 times G velocity sedimentation. Fractions were tested for their ability to restore the adoptive antibody response of irradiated hosts to horse spleen ferritin. A constant source of T cells (small numbers of unfractionated thoracic duct cells) was added to each fraction in order to monitor the B cell activity of the latter. Although large and small cell fractions of the spleen showed restorative activity, only the small cell fractions of the thoracic duct lymph showed activity. The turnover rate of the spleen cell fractions was determined by treating donors with high specific-activity 3H-thymidine for 48 hr before splenectomy. Rapidly dividing cells are preferentially killed by this treatment. The results suggest that a considerable proportion of large, intermediate, and small virgin B cells turn over within 48 hr. The cell surface of the various spleen cell fractions was examined for the presence of immunoglobulin (Ig) and a receptor for complement. The percentage of Ig-bearing cells in the large cell fractions was similar to the percentage of cells bearing IgM and a receptor for complement. However, the majority of Ig-bearing cells in the small cell fractions did not show the latter two surface markers. Experiments with the fluorescence-activated cell sorter showed that the large functionally active B cells bore surface IgM. The experimental findings suggest that there are subpopulations of virgin B cells in the spleen of the adult rat which differ with respect to size, migration pattern, turnover rate, and cell surface characteristics. The relationship of these cells to one another is discussed in the framework of an antigen-independent model of B cell maturation in the rat.

Animals↗
PubMed1979

The development of the hippocampus and dentate gyrus in normal and reeler mice.

The histogenesis, the time of origin and the pattern of migration of the cells in the hippocampus and dentate gyrus, have been studied in normal and reeler mice. The earliest indication of a defect in the reeler hippocampus is seen on the fifteenth embryonic day (E15) which is at least 24 hours after the first indication of a defect in the neocortex. It is not until E18, that the dentate gyrus shows signs of its incipient abnormality. It appears then, that in both the hippocampus and the dentate gyrus the gene defect first manifests itself at the stage at which the definitive cellular layers are assembled. Experiments involving the injection of 3H-thymidine (3H-TdR) at different developmental stages have confirmed that the site and rate of cellular proliferation in the reeler hippocampus and dentate gyrus are normal, as is the initial pattern of cell migration. However, in the reeler dentate gyrus, most postnatal cell proliferation occurs ectopically and in the hippocampus the normal "inside-out" sequence of neurogenesis is reversed, the earliest pyramidal cells generated coming to lie superficially within the stratum pyramidale and the later formed cells being added at progressively deeper levels. There is no discernible gradient in the time of origin of the granule cells in the radial dimension of the reeler dentate gyrus, whereas there is an obvious "outside-in" gradient in the normal animal. The characteristic gradients in cell proliferation seen in the transverse and longitudinal dimensions of the normal dentate gyrus are, however, also evident in the reeler mouse. Taken together, these observations suggest that the reeler gene exerts its effect on neuronal position only in the radial dimension, and does so at a stage of development subsequent to the proliferation and initial migration of the relevant neurons. Timm's sulfide silver preparations indicate that the characteristic staining patterns seen in the dentate gyrus and hippocampus appear at the same time, and mature at the same rate in normal and reeler mice.

Animals↗
PubMed2025

SimHumanity: Using SLiM 5.0 to run whole-genome simulations of human evolution.

The reconstruction of human evolutionary history has undergone repeated advances, each made possible by methodological innovations. In recent decades, genetic and genomic data played a central role in the reconstruction of major evolutionary events such as the out-of-Africa migration, and genetic simulations of human evolutionary history have come to play a major role in testing more specific hypotheses including proposed patterns of migration and admixture with archaic hominins. Increasing computational power has allowed human evolutionary history to be modeled at ever-larger scales, but simulations that encompass the complete human genome, including sex chromosomes and mitochondrial DNA, have been difficult due to the lack of support for whole-genome models in commonly used evolutionary simulation frameworks. With the recent introduction of SLiM 5 such simulations are now straightforward to construct, allowing the easy simulation of humans at whole-genome scale under different demographic models and evolutionary dynamics. We here present three versions of a reusable, customizable, open-source SLiM 5 model for simulating the molecular evolution of the full human genome. We also show some simple analyses of results from the model, to illustrate its utility. We hope this model, which we have nicknamed "SimHumanity" in jest, will facilitate further progress in the field of human evolutionary simulations.

SLiM↗
PubMed2025

Migration strategies, connectivity and corridor features of the partial migrant little bustard (Tetrax tetrax) across the Iberian Peninsula.

The study of migration ecology is crucial for understanding the factors and pressures affecting migratory species. Here, we studied the migratory ecology of the little bustard (Tetrax tetrax), a steppe bird that has suffered a sharp decline over recent decades, mainly due to agricultural intensification. Using 105 adult birds tagged across the main Iberian regions where the species is present (Alentejo, Extremadura, Ebro Valley, Northern Plateau, Southern Plateau and Guadalquivir Valley), we analysed the ratio of migratory and resident birds in each population and assessed their connectivity during the three main migratory periods (summer, winter and pre-breeding). Additionally, we describe the features of the migrations recorded in terms of length, duration and day period. Our results corroborate that little bustards can be considered partial migrants across Iberia, although the proportion of residents versus migrants varied between populations: the Alentejo (94.74%) and Northern Plateau (93.75%) had the highest proportion of migrants, followed by Guadalquivir Valley (81.82%), Extremadura (65.38%), Southern Plateau (55.56%) and Ebro Valley (25.93%). Migratory connectivity varied between periods: the pre-breeding and summering migrations showed a trend to move northwards, while birds moved southwards for winter. Regarding the migratory corridors obtained from the 253 migrations identified, we found three main routes: one corridor that connects the Northern Plateau with the western part of the Southern Plateau and Extremadura, another one that connects the Southern Plateau, Extremadura, Alentejo and Guadalquivir Valley, and one corridor that concentrates migrations within the Ebro Valley, and between the Ebro Valley and the Southern Plateau. Finally, analyses showed that little bustards migrate at night through areas dominated by herbaceous cover (avoiding tree-covered land and water bodies) and of low elevation and terrain roughness. Our results highlight the importance of developing an international and inter-regional conservation strategy to protect not only the breeding and wintering quarters, but also this endangered species' migratory corridors, thus supporting the viability of the metapopulation.

Brownian bridge kernel↗
PubMed1977

Association of a low molecular weight helper factor(s) with thymocyte proliferative activity.

Human mixed leukocyte supernatants contain thymocyte proliferative activity (TPA) and a low m.w. helper factor, designated HP-1, which is capable of partially restoring the antibody response of T-cell-deficient adherent murine spleen cells to the thymic-dependent antigen, SRC. TPA and HP-1 appear to have a comparable m.w. (14,000 to 14,500 daltons) by Sephadex gel filtration column chromatography. Furthermore, HP-1 and TPA exhibit similar patterns of heterogeneity on DEAE-cellulose chromatography, elute together on CM-cellulose chromatography, and manifest identical patterns of migration on polyacrylamide gel electrophoresis. These data suggest that the TPA and HP-1 activities reside in either the same molecule(s) or in different molecules with identical charge/mass ratios. Furthermore, the results support the hypothesis that the helper activity of HP-1 is derived from its capacity to activate T and/or pre-T cells.

Animals↗
PubMed1977

Migrating myoelectrical complex of the small intestine. An intrinsic activity mediated by the vagus.

In healthy conscious parenterally fed dogs and in sheep on their usual diet, the basic motor profile on the small intestine consists of recurring cycles of action potential activity. This cyclic pattern called the migrating myoelectrical complex comprises two distinct phases termed irregular, and regular spiking activity and is followed by a period of quiescence. The pattern persists after bilateral transthoracic vagotomy and in animals rendered diabetic by alloxan. In dogs, feeding disrupts the migrating myoelectrical complex pattern by obliterating the phases of regular spiking activity and quiescence for 6 to 12 hr, depending upon the amount of dry matter intake. After vagotomy a latency in the disruption of the migrating myoelectrical complex pattern with feeding occurs and in vagotomized dogs rendered diabetic, the duration of disruption is strongly reduced. In sheep, the duration of irregular spiking activity of a jejunal segment increased or decreased corresponding to the bulk of digesta, a phenomenon damped after vagotomy. The results indicate that the role of the vagus is limited to (1) prompting prandial disruption, which is then maintained by hormonal effect; (2) regulating the irregular spiking activity duration in relation to the bulk of digesta.

Action Potentials↗
PubMed1979

Rapid change of chromomeric and pairing patterns of polytene chromosome tips in D. melanogaster: migration of polytene-nonpolytene transition zone?

The high variability of chromomeric patterns in near-terminal regions of polytene chromosome arms has been explored in a number of races, strains and hybrids of Drosophila melanogaster. Traditional explanations for tip differences between strains (differential compaction of chromatin, somatic or germinal deletion) are examined and, in the light of the reported observations, rejected. The range of polytene tip variability and rates of change in wild races are greater than has been supposed: strains formerly considered to be terminally deleted appear to gain terminal bands; others, formerly considered normal, appear to have lost them. Strains with high cell-to-cell tip variability are also described. Cell-to-cell variations, as well as much of the observed rapid changes in tip appearance, are probably due to heritable differences in the location of an abrupt transition zone between polytene and nonpolytene chromatin. A quantitative relationship between the amount of certain subterminal bands present and the frequency of tip association of nonhomologous chromosomes is shown and its possible significance for chromosome is shown and its possible for chromosome pairing discussed.

Animals↗
PubMed1977

[Migration of transformed fibroblast-like cells on substrates with patterned relief-work (quantitative study)].

The quantitative estimation of migration ability of transformed fibroblast-like cells of various epecies (mouse , rat, hamster, man) cultured on the substratum with grooves of various depth (5--40 mcm) was performed. Of the majority of 11 cell lines examined, a reduced migration capacity up to its total disappeanance was registered as compared with the homological normal embryonic cells. A more pronounced reduction of migration ability was shown in mouse and rat transformed cells and less in human ones. Migration of transformed hamster cells was just equal to the weak migrtion of normal hamster embryonis cells. These data shown a weak response of the analysed transformed cells to the relief of underlying substratum.

Animals↗
PubMed1975

Characterization of human alpha-galactosidase A and B before and after neuraminidase treatment.

It has been previously reported that following neuraminidase treatment alpha-galactosidase A is converted into the B form, as revealed by electrophoresis. By a variety of techniques such as isoelectrofocusing, DEAE-chromatography and by enzyme kinetic parameters, no conversion of alpha-galactosidase A into B, or the reverse, could be detected after neuraminidase treatment. Only an apparent transformation of alpha-galactosidase A into B was revealed by Cellogel electrophoresis. In addition, a discrepancy was noticed between the pattern of electrophoretic migration on starch gel and Cellogel and the net electrical charges of the two alpha-galactosidases as deduced by isoelectrofocusing and DEAE-cellulose. Neuraminidase treatment did not affect the activity of alpha-galactosidase A towards the natural substrate, ceramidetrihexoside, but the activity of alpha-galactosidase B decreased by about 30% under the same conditions. The two forms of alpha-galactosidases A and B used in this study were extensively purified by classical procedures.

Animals↗
PubMed1975

Effect of antithymocyte globulin pretreatment on immunological reconstitution of lethally irradiated animals.

Clinical and experimental studies have shown that antithymocyte globulin pretreatment will reduce the severity of the graft-versus-host reaction after allogeneic bone marrow transplantation. To determine whether this was attributable to a persisting cytotoxic factor in the recipients' sera or a result of the "masking" of foreign antigens by the antihymocyte globulin, an experimental schema utilizing a syngeneic transfer of immunocompetent cells was devised. Lethally irradiated mice that had been pretreated with rabbit antimouse thymocyte globulin (RAMTG) were injected with syngeneic spleen or bone marrow cells and their immunological competence was measured by the response to a test antigen, sheep red blood cells. It was found that such pretreatment had an adverse effect on the immunological potential of the infused cells. Thus, the plaque-forming cell response to sheep red blood cell antigen in RAMTG-pretreated recipients injected with spleen cells was reduced when compared to the saline or normal rabbit globulin-treated controls. The immunological recovery of lethally irradiated animals protected with syngeneic bone marrow was also delayed, but not permanently impaired, when the recipients had been pretreated with RAMTG. These effects were evident although the spleen or bone marrow cells had been injected into the RAMTG-pretreated recipients at a time when their sera were devoid of any cytotoxic antibodies. It is speculated that two mechanisms may contribute to this alteration in immune function of the infused cells: (1) that RAMTG exists in the serum in amounts sufficient to attach to receptor sites on lymphocytes or their precursors but insufficient to kill the cells, interfering with the antigen recognition mechanism or migration and homing patterns, or (2) that the RAMTG treatment creates a temporary defect in the microenvironment of the spleen and other hemopoietic tissues, thereby affecting the transplantation and proliferation kinetics of the infused cells.

Animals↗
PubMed1977

The role of cell-mediated immunity in the induction of inflammatory responses. Parke-Davis Award Lecture, 1977.

Reactions of cell-mediated immunity fall into two broad categories: those that involve direct participation of intact lymphocytes in the effector mechanism of the reaction and those that involve mediation by soluble lymphocyte-derived factors known as lymphokines. The first kind of reaction is essentially limited to lymphocyte-dependent cytotoxicity, although certain aspects of T cell-B cell cooperation may fall into this category as well. The second category appears to comprise the bulk of the so-called cell-mediated immune response and provides a link between this system and the inflammatory system. Various lymphokines have been shown to exert profound influence upon inflammatory cell metabolism, cell surface properties, patterns of cell migration, and the activation of cells for various biologic activities involved in host defense. Although substantial information is now available about various physicochemical as well as biologic properties of lymphokines, purification and characterization data are as yet too incomplete to allow us to ascribe all of these activities to discrete mediator molecules. Current work involving the development of antibody-based techniques for mediator assay may shed light on this issue. Information on the kinds of cells capable of lymphokine production is now available. Contrary to prior expectation, T cells are not unique in their capacity for lymphokine production. Under appropriate circumstances, B cells and even nonlymphoid cells can do so as well. The unique property of lymphocytes in this regard appears to relate to their ability to respond to certain specialized signals such as specific antigen or an appropriate mitogen. Mediator production per se may represent a general biologic phenomenon. Although lymphokines have been defined mainly in terms of in vitro assays, early speculations about their in vivo importance are proving correct. Evidence for the role of lymphokines comes from studies involving detection of lymphokines in tissues, studies involving injection of exogenous lymphokines, and studies involving suppression of in vivo reactions by various techniques. The use of antilymphokine antibodies has proven useful in the latter kinds of experiments. Work in many laboratories is beginning to relate these findings to clinically relevant situations. A major unsolved problem relates to the regulation and control of lymphokine production and activity. At present only a limited body of information is available on this point. This is a potentially fruitful area for future investigation since it may provide techniques for manipulating the immune system in ways that are clinically useful.

Animals↗
PubMed1979

Lipoprotein abnormalities in cholestasis. I. Electrophoretic and ultracentrifugal analyses.

The alterations of lipid composition in sera of patients with liver diseases, particularly intrahepatic cholestasis and biliary obstruction, were studied by ultracentrifugation and polyacrylamide-gel disc-electrophoresis of lipoproteins and apoproteins. The elevation of serum cholesterol in intrahepatic cholestasis was greater than in biliary obstruction. The appearance of lipoprotein X in obstructive disease accounted for most of the increased cholesterol. The level of non-lipoprotein X cholesterol in intrahepatic cholestasis was significantly elevated, this being in part ascribed to the appearance of a new class of cholestatic lipoprotein, Slow-migrating HDL. The electrophoretic pattern of lipoprotein in cholestasis was generally characterized by a decrease in alpha band intensity and, in some types of cholestasis, by the appearance of Slow-migrating HDL. In addition, other abnormal lipoproteins exhibiting the characteristics of triglyceride-rich LDL (LP-Y), LP-X-like HDL and LDL-like HDL were found in some cases of intrahepatic cholestasis and biliary obstruction.

Apolipoproteins↗