The effect of interleukin-2 on experimental emotional hypertension]

The effect of human recombinant interleukin-2, one of the potent mediators of the immune system, on the course of emotional hypertension in non-linear white rats has been investigated. A significant and prolonged hypotensive action of a single injection of interleukin-2 in hypertensive rats has been revealed. The data obtained can be a new evidence of participation of immune system in the development of hypertension in experimental animals.     

A new affinity reagent for the site-specific, covalent attachment of DNA to active-site nucleophiles: application to the EcoRI and RsrI restriction and modification enzymes.

A modified oligodeoxyribonucleotide duplex containing an unnatural internucleotide trisubstituted 3' to 5' pyrophosphate bond in one strand [5'(oligo1)3'-P(OCH3)P-5'(oligo2) 3'] reacts with nucleophiles in aqueous media by acting as a phosphorylating affinity reagent. When interacted with a protein, a portion of the oligonucleotide [--P-5'(oligo2)3'] becomes attached to an amino acid nucleophilic group through a phosphate of the O-methyl-modified pyrophosphate linkage. We demonstrate the affinity labeling of nucleophilic groups at the active sites of the EcoRI and RsrI restriction and modification enzymes with an oligodeoxyribonucleotide duplex containing a modified scissile bond in the EcoRI recognition site. With the EcoRI and RsrI endonucleases in molar excess approximately 1% of the oligonucleotide becomes attached to the protein, and with the companion methyltransferases the yield approaches 40% for the EcoRI enzyme and 30% for the RsrI methyltransferase. Crosslinking proceeds only upon formation of a sequence-specific enzyme-DNA complex, and generates a covalent bond between the 3'-phosphate of the modified pyrophosphate in the substrate and a nucleophilic group at the active site of the enzyme. The reaction results in the elimination of an oligodeoxyribonucleotide remnant that contains the 3'-O-methylphosphate [5'(oligo1)3'-P(OCH3)] derived from the modified phosphate of the pyrophosphate linkage. Hydrolysis properties of the covalent protein-DNA adducts indicate that phosphoamide (P-N) bonds are formed with the EcoRI endonuclease and methyltransferase.     

Immortalization of hypothalamic GnRH neurons by genetically targeted tumorigenesis

By genetically targeting tumorigenesis to specific hypothalamic neurons in transgenic mice using the promoter region of the gonadotropin-releasing hormone (GnRH) gene to express the SV40 T-antigen oncogene, we have produced neuronal tumors and developed clonal, differentiated, neurosecretory cell lines. These cells extend neurites, express the endogenous mouse GnRH mRNA, release GnRH in response to depolarization, have regulatable fast Na+ channels found in neurons, and express neuronal, but not glial, cell markers. These immortalized cells will provide an invaluable model system for study of hypothalamic neurosecretory neurons that regulate reproduction. Significantly, their derivation demonstrates the feasibility of immortalizing differentiated neurons by targeting tumorigenesis in transgenic mice to specific neurons of the CNS.     

Response of Picea rubens seedlings to intermittent mist varying in acidity, and in concentrations of sulfur-, and nitrogen-containing pollutants

Two-year-old red spruce seedlings ( Picea rubens Sarg .) growing in. field chambers were repeatedly exposed to acidic mist with a factorial combination of 3 fluctuating levels of acidity: median pH values of 3.0 (range of 2.5 to 3.5), 3.5 (range of 3.0 to 4.0), and 4.0 (range of 3.5 to 4.5). and 3 ion compositions: sulfate. nitrate and ammonium, and a combination of all 3 ions. The experiment was performed during the growing season over a period of 3.5 months. Mist exposures were intermittent with 5 wet-dry cycles for each 16-h overnight exposure period, Foliar necrosis occurred on seedlings treated with the most acidic mist and was most severe when the mist contained sulfate. At a median pH of 3.5, a value close to that of cloud water occurring in the eastern United States, injury developed with sulfuric acid mist, but not. when the mist contained nitric acid. The combination of high acidity and sulfate significantly decreased volume of aboveground tissues, while high acidity and nitrate increased volume. Root and needle dry weights were not affected. However, high acidity of mist was associated with increased leader shoot length. These results indicate, that there is a risk of foliar injury and changes in growth of red spruce with cloud water at a median acidity of pH 3.5 or below, especially when there are high concentrations of sulfate and low concentrations of nitrate.     

Persistence of Epstein-Barr Virus in Self-Reactive Memory B Cells

Epstein-Barr virus infection has been epidemiologically associated with the development of multiple autoimmune diseases, particularly systemic lupus erythematosus and multiple sclerosis. Currently, there is no known mechanism that can account for these associations. The germinal-center (GC) model of EBV infection and persistence proposes that EBV gains access to the memory B cell compartment via GC reactions by driving infected cells to differentiate using the virus-encoded LMP1 and LMP2a proteins, which act as functional homologues of CD40 and the B cell receptor, respectively. The ability of LMP2a, when expressed in mice, to allow escape of autoreactive B cells suggests that it could perform a similar role in infected GC B cells, permitting the survival of potentially pathogenic autoreactive B cells. To test this hypothesis, we cloned and expressed antibodies from EBV⁺ and EBV⁻ memory B cells present during acute infection and profiled their self- and polyreactivity. We find that EBV does persist within self- and polyreactive B cells but find no evidence that it favors the survival of pathogenic autoreactive B cells. On the contrary, EBV⁺ memory B cells express lower levels of self-reactive and especially polyreactive antibodies than their uninfected counterparts do. Our work suggests that EBV has only a modest effect on the GC process, which allows it to access and persist within a subtly unique niche of the memory compartment characterized by relatively low levels of self- and polyreactivity. We suggest that this might reflect an active process where EBV and its human host have coevolved so as to minimize the virus''s potential to contribute to autoimmune disease.     

Genetic Control of Spermatogenesis and Sex Determination in Mammals

The material was analyzed on the main problems of genetics of mammalian spermatogenesis, sex determination, its reversion and other defects from the standpoint of current cytological and molecular-genetic concepts of functional activity of the parental genomes after fertilization and behavior of their chromosomes at the early embryonic stages. On the basis of this analysis, a hypothesis has been proposed, which explains a high percentage (50% or more) of early embryonic mortality in placental mammals under the conditions of natural and extracorporeal fertilization, as well as regular appearance of defects in the course of natural sex determination, including the appearance of representatives of both sex minorities. We do not make pretense to comprehensive and deep analysis of male gametogenesis and sex determination in mammals.