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Malolina E.A.,RAS D. I. Ivanovsky Institute of Virology | Kulibin A.Y.,Russian Academy of Sciences | Naumenko V.A.,RAS D. I. Ivanovsky Institute of Virology | Gushchina E.A.,RAS D. I. Ivanovsky Institute of Virology | And 2 more authors.
International Journal of Experimental Pathology | Year: 2014

Summary: This study aimed to establish the influence of herpes simplex virus (HSV) on testis morphology and germ cell development using a model of ascending urogenital HSV infection in mice. Adult C57BL/6J mice were inoculated with 100 plaque-forming units of HSV1 in rete testis. Viral proteins and HSV DNA were detected from 3 days postinoculation (DPI), while capsids and virions could be visualized at 6 DPI. Infectious activity of HSV was revealed by rapid culture method in testes from 3 to 14 DPI, and virus DNA by PCR - from 3 to 100 DPI. Germ and Sertoli cells were infected during the early stages of the infection, whereas interstitial cells only occasionally contained the virus at 21 and 45 DPI. Microscopic analysis revealed severe degeneration of the germinal epithelium in the infected testes. By 21 DPI, testes became atrophic and most Sertoli cells were destroyed. No testicular regeneration and no spermatozoa in the epididymis were observed at 45 and 100 DPI. From 3 DPI, inflammatory cells accumulated in the interstitium between damaged tubules; a significant increase in the number of CD4+, CD8+ T lymphocytes and F4/80+ cells was observed in the infected testes. This study shows that in the case of HSV retrograde ascent into seminiferous tubules, the acute viral infection results in irreversible atrophy of the germinal epithelium, orchitis and infertility. These results may be used to further study viral orchitis and the influence of HSV on spermatogenesis and male fertility. © 2014 International Journal of Experimental Pathology. Source


Polzikov M.A.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Kordyukova M.Y.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Zavalishina L.E.,Gertsen Research Institute of Oncology | Magoulas C.,Queen Mary, University of London | Zatsepina O.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry
Hybridoma | Year: 2012

SURF-6 is an evolutionarily conserved nucleolar protein that is important for cell viability; however, its function in mammals still remains uncertain. The aim of this study is to generate monoclonal antibodies to human SURF-6 protein suitable for fundamental and biomedical research. The full-size human SURF-6 was expressed as a recombinant GST-fusion protein and used as an antigen to generate monoclonal antibodies, S79 and S148, specific for SURF-6. The monoclonal antibody produced by hybridoma clone S79 specifically recognizes endogenous SURF-6 by Western and immunofluorescence analyses in various cultured human cells, and by immunohistochemistry in paraffin-embedded sections of human breast cancer samples. Moreover, S79 immunoprecipitates protein complexes containing SURF-6 from HeLa cells extracts. The antibody S79 recognizes SURF-6 only in human cells; however, the antibody produced by hybridoma clone S148 can detect SURF-6 of human and mouse origin. Monoclonal antibodies to the nucleolar protein SURF-6 described in this work can be a useful tool for studies of ribosome biogenesis in normal and cancer cells. © 2012, Mary Ann Liebert, Inc. Source


Gol'Dberg M.A.,RAS Institute of Metallurgy | Smirnov V.V.,RAS Institute of Metallurgy | Kutsev S.V.,RAS Institute of Metallurgy | Shibaeva T.V.,RAS Institute of Metallurgy | And 4 more authors.
Inorganic Materials | Year: 2010

Hydroxyapatite/calcium carbonate (CC) composite powders containing up to 50 wt % CO 3 2-, have been prepared via precipitation from aqueous solutions. According to chemical analysis data, the CO 3 2- content of the powders coincides with the intended one over the entire composition range studied. With increasing CO 3 2- content, the specific surface area of the powders decreases because of the formation and growth of large needlelike CC crystals up to 1.5 μm in size. The addition of low-melting-point alkali carbonates to the starting powder mixture reduces the sintering temperature of the powders to below 720°C, thereby preventing the thermal decomposition of CC. The highest bending strength, up to 76 MPa, is offered by the materials with the lowest CC content, about 20 wt %, and an average crystal size of 100 nm. The solubility of the composites gradually increases with CC content. In vitro experiments with a human fibroblast model (MTT test) demonstrate that the composites have zero cytotoxicity. © 2010 Pleiades Publishing, Ltd. Source


Balabushevich N.G.,Moscow State University | Borzenkova N.V.,Moscow State University | Izumrudov V.A.,Moscow State University | Larionova N.I.,Moscow State University | And 3 more authors.
Applied Biochemistry and Microbiology | Year: 2014

Suspensions of insoluble polyelectrolyte complexes of dextran sulfate (DS) of different molecular masses with lactoferrin (LF) have been fabricated and characterized. The encapsulation efficiency of LF and DS in a complex at pH 3.0 and 4.0 was assessed, and particles were characterized by their sizes and ζ-potential. The complexes formed at pH 3.0 differed by a higher stability level. The interaction with DS resulted in a twofold decrease in the antioxidant activity of LF, although the formation of complexes was not accompanied by conformational changes in LF molecules according to IR-spectrometry data. Microencapsulation was carried out by treating the suspensions with negatively charged LF-DS complexes with protamine and chitosane solutions with different molecular masses. The composition, size, and the ζ-potential of interaction products were assessed which allowed us to select the conditions for the preparation of pH-sensitive polyelectrolyte microparticles loaded with LF which would be able to gradually release glycoprotein under conditions that model the passage through the gastrointestinal tract of humans. These data indicate that this approach is promising for the creation of pH-sensitive biopolyelectrolytes suitable for oral administration of LF to target cells. © 2014 Pleiades Publishing, Inc. Source


Akhidova E.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Volkova T.D.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Koroev D.O.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Kim Ya.S.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | And 6 more authors.
Russian Journal of Bioorganic Chemistry | Year: 2010

Survivin, an endogenous protein, is a promising marker for the cancer diagnosis. The aim of the present work was to obtain antibodies specific to survivin and capable of detecting this protein in tumor tissues. Four peptides corresponding to fragments (1-22), (54-74), (80-88)-(153-165), and (118-144) of the survivin-2B sequence were selected and synthesized. Rabbits were immunized with the synthetic peptides. It has been shown that all peptides in a free state, without conjugation with a high-molecular-weight carrier, stimulate the production of antibodies capable of binding with recombinant survivin. Antipeptide antibodies were isolated from sera and their performance in the immunohistochemical detection of survivin in human tumor tissues was studied. It was shown that only antibodies to the (80-88)-(153-165) peptide bind to the survivin present in breast and bladder tumors. The ability of antibodies to this peptide to detect survivin in tumor tissue lysates was demonstrated by immunoblotting. The part of the sequence targeted by the antibodies against the (80-88)-(153-165) peptide was localized using truncated peptide fragments. © 2010 Pleiades Publishing, Ltd. Source

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