Effect of antihepatocytotoxic serum on DNA synthesis in the rat

Incorporation of thymidine-³H into parenchymatous and reticulo-endothelial cells of the liver was studied autoradiographically in adult female rats treated with small doses (0.06 g/100 g body weight per injection) of antihepatocytotoxic serum (AHTS), the -globulin isolated from it (AHTS), and the -globulin fraction of normal rabbit serum (NRS) to intact animals and to rats with liver damage caused by carbon tetrachloride (CCl₄). Following injection of AHTS and, to a lesser degree, of AHTS into intact animals the index of labeled nuclei of both the parenchymatous and the reticulo-endothelial cells was increased. When given after preliminary CCl₄ administration, AHTS stimulated reparative regeneration. The action of AHTS took place in phases: A period of increase in the index of labeled nuclei was followed by a period of decrease, and this again was followed by a fresh period of stimulation of proliferative processes.Department of Immunology and Cytotoxic Sera, A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. (Presented by Academician of the Academy of Medical Sciences of the USSR N. N. Gorev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 7, pp. 75–78, July, 1978.     

Structure of the liver as a possible factor in the regulation of α-fetoprotein synthesis

During regeneration of the mouse liver after poisoning with CCl₄ and paracetamol, hepatocytes containing -fetoprotein (FP) lose their membrane antigen that served as marker for biliary capilaries. Both during regeneration and in early postnatal development, cessation of FP synthesis by the cells coincides with the appearance of an antigen marking the biliary capillaries on their surface. It is suggested that cessation of FP synthesis is the result of establishment of a system of contacts characteristic of the definitive hepatic column.Laboratory of Immunochemistry and Diagnosis of Tumors, Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 6, pp. 576–579, June, 1979.     

A comparative immunohistological study of cerebellar enolases

Summary A comparative immunohistological study of the neurone-specific enolase and enolase, demonstrates the exclusive neuronal localization of enolase and its absence from glial cells. In contrast, enolase is located in astroglial cells. The validity of enolase as a neuronal marker and enolase as an astrocytic marker, is confirmed both by a double labelling technique, using antibodies to and to revealed with fluorescence or peroxidase in the same tissue sections, and by immunoelectronmicroscopy.     

Isoelectric focusing and immunochemical analysis of proteins of the “Diaferm-3” antitetanus serum

The isoelectric points (PIs) of proteins present in the Diaferm-3 antitetanus serum, a mixture of partially purified [F'ab]₂-fragments of native molecules of antitetanus immunoglobulins, their antitoxic activity, and their immunochemical properties were investigated by isoelectric focusing in a pH gradient of ampholines. The PI values and electrophoretic mobility of the preparation were compared. Proteins with antitoxic activity were shown to have PI values within the range 9.1–5.15, and from their electrophoretic mobility they can be classed as G- and -globulins. No antitoxic activity was found in macroglobulins with the mobility of -globulins.Laboratory of Pathophysiology of Toxicoinfections, Institute of Normal and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. M. Chernukh.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 78, No. 10, pp. 41–45, October, 1974.     

Activation of atrial muscarinic K+ channels by low concentrations ofβγ subunits of rat brain G protein

Effects of G protein subunits from rat brain on cardiac K⁺ channel was examined in single atrial cells of guinea-pig, using patch clamp techniques. We found that 10 pM concentration of rat brain subunits preparation could activate the atrial muscarine receptor-gated K⁺ channel (I_K.ACh). Neither the detergent, CHAPS, used to suspend nor the boiled preparation activated I_K.ACh. Furthermore, preincubation of subunits preparation in Mg²⁺-free solution, which easily inactivated -GTP-S, did not affect -activation of I_K.ACh. We concluded, therefore, that subunits themselves can activate I_K.ACh.Supported by the grants from the Ministry of Education, Culture and Science of Japan and from the Calcium Signal Workshop on Cardiovascular Systems     

Aggravation of injury to liver lysosomes of rats with CCl4 hepatitis by preliminary prolonged administration of chlorpromazine

Preliminary prolonged administration of chlorpromazine (5 mg/kg for three weeks) aggravated the injury to liver lysosomes of rats with acute CCl₄ hepatitis. Similar marked changes were observed in lysosomes sedimented with heavy and light mitochondrial fractions.Central Scientific-Research Laboratory and Department of Psychiatry, Novosibirsk Medical Institute. (Presented by Academician, of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 1, pp. 38–41, January, 1977.     

Cells with fragmented nuclei in ascites hepatoma 22A and their role in proliferation

During aging of an ascites hepatoma 22A (AH22A) the number of cells with fragmented nuclei (especially multilobate) increases: in an AH22A aged 6 days they numbered 15±9.3, in a tumor aged 14 days 196±53, and in a delayed tumor aged 18 days 453±51. The main method of formation of fragmented nuclei is by amitosis. Approximately 150 and 170 of cells with fragmented nuclei in a 14- and 18-day old AH22A were in the reversible resting R₁ stage (or in a very protracted G₁-period, extending over 4 days), whereas the remaining 50 and 230 of cells respectively had left the mitotic cycle irreversibly and were evidently undergoing involution, which takes place more rapidly during passage-stimulated division.Laboratory of Chemical Factors of Regulation of Growth and Cell Division, Institute of Biological and Medical Chemistry, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR V. N. Orekhovich.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 10, pp. 445–447, October, 1979.     

GABA<Subscript>A</Subscript> receptor subunit expression in the guinea pig vestibular nucleus complex during the development of vestibular compensation

The aim of this experiment was to investigate whether vestibular compensation following unilateral vestibular deafferentation (UVD) is associated with changes in the expression of GABA_A receptor subunits in the guinea pig vestibular nuclear complex (VNC) at 2, 10, and 30 h post-surgery. Using Western blotting, the ₁ and ₂ subunits (but not the ₂ subunit) were detected in the VNC of labyrinthine-intact animals. However, there were no significant differences in the protein expression of the ₁ and ₂ subunits within the ipsilateral or contralateral VNC at any time post-UVD compared to sham and anesthetic controls. Furthermore, UVD did not induce the expression of the ₂ protein. These results suggest that vestibular compensation in guinea pig, as in the rat, is not associated with changes in the protein levels of the GABA_A receptor subunits ₁, ₂, and ₂ in the VNC. However, a limitation of this study is that the Western blotting technique can detect only changes that are larger than 30% and therefore small changes cannot be excluded.     

Über Veränderungen im Kohlenhydratgehalt der Serumproteine bei Kala-Azar

Zusammenfassung Die elektrophoretisch isolierbaren Fraktionen der Serumproteine zeigten bei Kindern, die an Kala-Azar erkrankt waren, charakteristische Veränderungen. Der in den Zellen des Reticuloendothels lebende Parasit verursacht eine Vermehrung der ₁- und ₂-Globulinfraktion bei gleichzeitiger prozentueller Verminderung des Kohlenhydratgehaltes dieser Fraktionen. Die -Globulinfraktion ist dagegen nicht nur in ihrer Menge, sondern auch in ihrem Kohlenhydratgehalt vermehrt. Fast des an die Serumproteine gebundenen Kohlenhydrats ist bei den Kala-Azar-Fällen in den prosthetischen Gruppen der -Globuline enthalten.     

Changes in the serum proteins of rats with radiation sickness

Summary Immunoelectrophoretic analysis of serum proteins in rats of the August strain irradiated with -rays in a dose of 630 R showed an increase in the heterogenity of ₂B- and ₂B-globulins with the appearance of electrophoretically faster or slower fractions retaining (₃B) or slightly changing (₂B) their antigenic properties. Beginning with the 7th day after irradiation the serum of irradiated animals is found to contain a new radiation antigen migrating during electrophoresis together with slow ₂-globulins and indicated by ₂x-protein. The protein fails to appear after irradiation of sera in vitro and is absent both in healthy and artificially infected nonirradiated animals.(Presented by Active Member of the Academy of Medical Sciences of the USSR N. N. Zhukov-Verezhnikov). Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 62, No. 10, pp. 57–61, October, 1966.     

Immunologic specificity,nature, and properties of suppressor cells induced in mixed lymphocyte cultures

After incubation of lymphocytes 1–5 days in allogeneic or syngeneic one-way mixed lymphocyte culture (MLC) suppressor cells blocking proliferation of 10 to 100 times as many syngeneic responding lymphocytes in fresh allogeneic MLC are generated. Neither allogeneic nor syngeneic suppressors have the properties either of B-cells or macrophages, both are resistant to mitomycin C, but their precursors are sensitive to cyclophosphamide. They differ in their properties both from T-killers and from T-suppressors induced in a lymphocyte monoculture. The allogeneic suppressors are heterogeneous T-cells as regards radiosensitivity and specificity. The formation of nonspecific T-suppressors is considerably reduced if embryonic calf serum in the medium is replaced by human or bovine serum. Syngeneic suppressors also are heterogeneous and differ from allogeneic in their greater resistance to anti-T-antibodies and irradiation, whereas their precursors are more resistant to hydrocortisone.Laboratory of Immunochemistry and Diagnosis of Tumors, Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 89, No. 3, pp. 315–318, March, 1980.     

Immunoelektrophoretische Studien an Myelomseren

Zusammenfassung 17 Seren von Kranken mit klinisch gesichertem Myelom wurden mit Hilfe der Papier- und der Immunoelektrophorese untersucht. Hierbei wurde jedes Serum 1. gegen ein Anti-Normalserum, 2. gegen ein Anti-Normalserum, aus welchem das -Globulin durch Absorption entfernt wurde, in 6 von den untersuchten Fällen auch 3. gegen das spezifische Antimyelomserum sowie 4. gegen das mit physiologischem -Globulin versetzte spezifische Antimyelomserum getestet.Es konnte festgestellt werden, daß sich Myelomfraktionen wie normale Globulinfraktionen verhalten, da sie I. bei Diffusion gegen Anti-Normalserum auspräcipitieren wie die entsprechenden physiologischen Fraktionen, und da II. der gegen die Myelomfraktionen gebildete Antikörper im entsprechenden Immunserum durch ein physiologisches Antigen — hier ein -Globulin — absorbiert wird, was darin zum Ausdruck kommt, daß Myelomfraktionen gegen ein Antimyelomserum nach Absorption durch -Globulinkeine Präcipitation mehr ergeben.Im einzelnen konnte die -Globulinfraktion von 9 papierelektrophoretisch reinen -Typen in 4 Fällen alseinheitliche, in weiteren 4 Fällen alsdoppelte und in einem Fall alsweitauslaufende Präcipitationslinie differenziert werden. Ferner waren zwei ₁-Myelome, ein ₂-, einM-Myelom, sowie drei ₂-Myelome immunoelektrophoretisch darstellbar. In einem papierelektrophoretisch normalen Serum konnte mit der hier angewandten immunologischen Methode eine im ₂-Bereich gelegene Präcipitationslinie nachgewiesen werden.Herrn Prof. Dr.H. Pette zum 70. Geburtstag.     

Isoenzymes of γ-glutamyl transpeptidase in liver diseases

Summary A new method for the separation of isoenzymes of-glutamyl-transpeptidase is described, using electrophoresis on acetate cellulose gel and a developing solution composed by-glutamyl-naphthylamide, and a colored diazonium compound.The method permits the separation of up to four different isoenzymes, which we called-GT₁,-GT₂,-GT₃,-GT₄, the first two showing an electrophoretic migration similar to that of ₁- and ₂-globulins and the other two to that of-globulins.The present technique has proved its usefulness in detecting isoenzymes in serum with values of total-glutamyl-transpeptidase higher than 80 U/L.The application of this method in 52 patients with different types of biliary obstruction and hepatocellular damage has shown that it provides new possibilities in differential diagnosis.     

Synergistic effects of tumour necrosis factor-α and interferon-γ on feline haemopoietic progenitors

Pathogenic mechanisms that underlie feline leukaemia virus subgroup-C (FeLV-C) induced erythroid aplasia are unknown. FeLV-C infection is associated with higher serum levels of interferon- (IFN-) and tumour necrosis factor- (TNF-), which may act synergistically to cause haemopoietic suppression. In the present studies, the synergistic effects of TNF- and IFN- on feline bone marrow progenitors in vitro were evaluated. Bone marrow mononuclear cells from specific-pathogen-free cats were exposed to TNF- (100 and 200 pg/ml) and IFN- (100 or 200 units/ml), alone or in combination, for 2 h before plating for clonal assays of colony forming units. Our results show that TNF- and IFN- in combination caused marked suppression of feline colony forming units-erythroid (CFU-E), burst forming units-erythroid (BFU-E), and colony forming units-fibroblasts (CFU-F), whereas colony forming units-granulocyte/macrophage (CFU-GM) were minimally affected. The same concentrations of TNF- and IFN- alone had minimal effects on CFU-E, BFU-E and CFU-F. These results suggest that TNF- and IFN- may play a significant role in regulating haemopoiesis in cats and may be involved in the pathogenesis of erythroid aplasia in cats infected with feline leukaemia virus.     

Acute and chronic inflammatory responses to local administration of recombinant IL-1α, IL-β, TNFα, IL-2 and Ifnγ in mice

The contention that cytokines are important mediators of inflammation prompted the present studies which were designed to compare acute and chronic pathological effects of locally-administered recombinant (r) IL-1, IL-1, TNF, IL-2 and Ifn. Acute (6 hr), resolving (48 hr) inflammation was induced by the following, in order of potency: rIL-1>rIL-1>rTNF>rIfn=BSA (control) following a single sc. injection. However, only rIL-1 and rIL-2 initiated and maintained chronic granulomatous reactions when delivered locally from a sc. ethylene vinyl acetate (EVA) slow-release polymer. The predominance of macrophages in EVA-rIL-1 lesions contrasted with the proliferative lymphoid granulomata induced by EVA-rIL-2 implants. These in vivo observations reinforce, the roles of both IL-1 and IL-2 as potent mediators of chronic immunoinflammatory disease.     

Effects of interferon-γ on cell differentiation and cytokine production of a human monoblast cell line,U937

The U937 cell, a human monoblast cell line, has been used as a model to study the function of human monocytes. We investigated the effects of interferon- (IFN-) on Superoxide anion (O ₂ ^– ) production, cell surface antigens, and cytokine production of U937 cells. IFN- treatment enhanced O ₂ ^– production of fMLP or PMA-stimulated U937 cells. IFN- increased the ratio of CD23 and CD 11b positive cells. The fluorescence intensity of CD14 and CD25 was enhanced by IFN- treatment. U937 cells produced IL-1, IL-1, IL-6, and TNF- by lipopolysaccharide (LPS) stimulation. IFN- treatment enhanced TNF- production, but decreased IL-6 production. These results suggest that IFN- differentiates U937 cells to monocytelike cells and it regulates the production systems of IL-6 and TNF- separately in U937 cells.     

α-Fetoprotein production in regenerating mouse liver synchronizedin vivo

Liver cells regenerating after CCl₄ poisoning were synchronizedin vivo by continuous administration of hydroxyurea. Accumulation of hepatocytes at the G₁-S phase boundary or in the S-phase did not affect the course of changes in the blood -fetoprotein (-FP) levels or the characteristic location of -FP for regenerating liver for 2 or 3 days after poisoning. -FP production began in the hepatocytes before their entry into the S-phase, and -FP was found in the cells also at different times after they had ended their mitotic cycle. No dependence of -FP synthesis on any particular phase of the mitotic cycle could be observed.Laboratory of Immunochemistry and Diagnosis of Tumors, Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 89, No. 3, pp. 346–349, March, 1980.     

Evidence for the presence of pro-γ-melanotropin,the NH2-terminal fragment of the corticotropin-β-lipotropin precursor,in corticotropin-producing tumours

Summary Five corticotropin-producing tumours were examined for peptides related to the corticotropin--lipotropin precursor. Two were basophil pituitary adenomas and three were bronchial carcinoids. The cells of the two pituitary adenomas stained with antisera against -endorphin and against pro--melanotropin, the NH₂-terminal fragment of the corticotropin--lipotropin precursor, but not with antisera against -melanotropin or -lipotropin. The corticotropin-storing tumor cells of the bronchial carcinoids stained with antisera against -endorphin, -lipotropin or pro--melanotropin. Only one of the three bronchial carcinoids contained cells reacting with the antiserum against -melanotropin. Although the two types of corticotropin-storing tumours (pituitary adenoma and bronchial carcinoid) differed with respect to -lipotropin content, the over-all picture indicates that the proteolytic processing of the corticotropin precursor proceeds along similar lines in tumour cells and in pituitary corticotrophs.An acetic acid extract of one of the bronchial tumours was subjected to gel chromatography and immunochemical analysis of material related to pro--melanotropin. The immunoreactive material displayed a considerable size heterogeneity, with the predominant components having a molecular weight larger than that of authentic pro--melanotropin.     

Density heterogeneities of hepatitis C virus in human sera due to the binding of β-lipoproteins and immunoglobulins

Heterogeneities in the density of hepatitis C virus RNA-carrying material (HCV-RNA-CM) found in human sera (1.03–1.20 g/cm³) are attributed to the binding of low-density lipoproteins and/or of IgG. In some sera HCV-RNA-CM seems to be nearly totally bound to -lipoproteins and cannot be precipitated by anti-IgG (); in others more than 95% of HCV-RNA-CM is bound to IgG and cannot be precipitated by anti--lipoprotein. Furthermore, there are sera from which HCV-RNA-CM can be completely be precipitated by either anti--lipoprotein or anti-IgG (), pointing to a binding of the two serum proteins to the same HCV-RNA-CM. There are other sera from which HCV-RNA-CM can be partially precipitated by the one or the other antiserum, leaving behind fractions, which are bound to -lipoprotein or to IgG. HCV-RNA-CM cannot be precipitated from some sera either by anti--lipoprotein or by anti-IgG ().     

Interferon β1a Treatment Modulates TH1 Expression in γδ + T Cells from Relapsing-Remitting Multiple Sclerosis Patients

A paradigm exists that multiple sclerosis is causally related to dysregulation of T_H1 inflammatory cytokines and T_H2 antiinflammatory cytokines. The cytokine source(s) that initiate the imbalances are unknown. In this study, , CD4, and CD8 T cell receptor-positive (TCR+) cells were isolated from the blood of 26 definitive relapsing-remitting multiple sclerosis patients prior to interferon -1a (IFN1a) therapy and following 8–10 weeks of this therapy. The bioactivities of interferon (IFN), interleukin 10 (IL10), and interleukin 12 (IL12) were determined. The concentrations of IFN, IL10, and IL12 from each cell type did not change significantly with IFN1a treatment. The IL10 secreted by TCR+ cells strongly correlated with the IL12 secreted by the same TCR+ cells, supporting the paradigm. Furthermore, IFN1a therapy decreased the TCR+ cell secretion of T_H1 cytokines after 8–10 weeks of therapy.