Growth-differentiation factor-8 (GDF-8) in the uterus: its identification and functional significance in the golden hamster

Transforming growth factor-beta superfamily regulates many aspects of reproduction in the female. We identified a novel member of this family, growth-differentiation factor 8 (GDF-8) in the 72 h post coital uterine fluid of the golden hamster by proteomic techniques. Uterine GDF-8 mRNA decreased as pregnancy progressed while its active protein peaked at 72 h post coitus (hpc) and thereafter stayed at a lower level. At 72 hpc, the GDF-8 transcript was localized to the endometrial epithelium while its protein accumulated in the stroma. Exogenous GDF-8 slowed down proliferation of primary cultures of uterine smooth muscle cells (SMC) and endometrial epithelial cells (EEC). In addition, GDF-8 attenuated the release of LIF (leukaemia inhibiting factor) by EEC. As for the embryo in culture, GDF-8 promoted proliferation of the trophotoderm (TM) and hatching but discouraged attachment. Our study suggests that GDF-8 could regulate the behavior of preimplantation embryos and fine-tune the physiology of uterine environment during pregnancy

Первичная медиастинальная В-крупноклеточная лимфома с редкой мутацией в гене ALK

Introduction. Primary mediastinal large B-cell lymphoma is an aggressive variant of lymphoma characterized by genetic heterogeneity. First-time therapy for primary mediastinal large B-cell lymphoma usually includes immunochemotherapy. However, a substantial proportion of patients do not respond to this therapy.Objective – to analyze clinical characteristics of primary refractory primary mediastinal large B-cell lymphoma taking into account the results of targeted next-generation sequencing (NGS).Materials and methods. A 22‑year-old patient with primary mediastinal large B-cell lymphoma who had not responded to immunochemotherapy was tested using targeted NGS for 77 genes.Results. We identified 2 rare mutations in the ALK gene with an unclear clinical value. According to the literature, these mutations are primarily found in solid tumors.Conclusion. Missense mutations identified in the ALK gene are presumably associated with the course of primary mediastinal large B-cell lymphoma, in particular, with primary refractory disease.Введение. Первичная медиастинальная В-крупноклеточная лимфома представляет собой агрессивный вариант лимфомы, характеризующийся генетической гетерогенностью. В 1-й линии лечения данной патологии принято проводить иммунохимиотерапию. Однако, несмотря на успехи в лечении первичной медиастинальной B-крупноклеточной лимфомы, есть большая группа пациентов, рефрактерных к терапии.Цель исследования – оценка особенностей клинического течения первично рефрактерной первичной медиастинальной В-крупноклеточной лимфомы с учетом данных таргетного секвенирования нового поколения (next generation sequencing, NGS).Материалы и методы. Пациентке, 22 лет, с первичной медиастинальной В-крупноклеточной лимфомой, у которой не наблюдалось эффекта от иммунохимиотерапии, выполнено таргетное секвенирование нового поколения с использованием панели из 77 генов.Результаты. В ходе таргетного секвенирования нового поколения выявлены 2 редких варианта мутаций в гене ALK с неясным клиническим значением. Согласно данным литературы они встречаются преимущественно в солидных опухолях.Заключение. Выявленные миссенс-мутации в гене ALK, возможно, связаны с особенностью течения первичной медиастинальной В-крупноклеточной лимфомы, в частности с первичной рефрактерностью к проводимому лечению

DNA-hydrolyzing activity of sIgA-abzymes of human milk is differentially inhibited by nucleoside triphosphates

It is known that human milk contains secretory immunoglobulines A (sIgA-abzymes) possessing affinity to mammalian DNA and capacity to cleave plasmid DNA. Regulation of DNA-hydrolyzing activity of those sIgA-abzymes is poorly studied. Here we investigate the effect of nucleoside triphosphates towards the DNase activity of sIgA-abzymes which were purified from human milk by a sequential chromatography on protein A-agarose, DEAE-Fractogel, and DNA-cellulose. By using DNA-hydrolysing assay, we revealed that 1 mM ATP and 1 mM dATP markedly reduced the cleavage of linear form of plasmid DNA by sIgA-abzymes, while the effect of GTP, CTP, TTP and dGTP was much weaker. dCTP and dTTP did not influence DNase activity of the sIgA-abzymes. Possible mechanisms of nucleotide-mediated inhibition of the DNase activity of sIgA-abzymes are discussed

Cytotoxic activity of anti-DNA slgA-antibodies from milk of healthy women

Earlier we have shown that immunoglobulin preparations derived from milk of healthy women by precipitation with 50% ammonium sulfate, possess toxic activity toward several transformed and tumor cells in vitro (Kit Y., and others, Biotechnology 2008). We hypothesized that cytotoxic effect was associated with the presence of cytotoxic anti-DNA sIgA-antibodies in the Ab preparations. To check this hypothesis, we have purified electrophoretic homogeneous anti-DNA sIgAs from human milk and stu­died their affect toward different tumor cells in vitro. Anti-DNA sIgAs were obtained by sequential chromatography on protein A-sepharose, DEAE-fractogel and DNA-cellulose columns. It was found that the sensitivity of studied cells to Ab changed in the order L929> L1210> Jurkat> Namalva. Annexin V – test , as well as DNA-fragmentation test showed that cell death under Abs action occurred by apoptosis

Characteristics of interaction of lectins with carbohydrate determinants of secretory immunoglobulin A (sIgA) isolated from colostrum of parturient women

Secretory immunoglobulin A (sIgA) contains glycoconjugates moieties which can be involved in humoral immunity providing antimicrobial defense. The aim of present work was to characterize carbohydrate moieties of sIgA isolated from human colostrum via their interaction with specific lectins. Preparations of sIgA were obtained from 23 colostrums samples of parturient women by 3-fold precipitation with in ammonium sulphate 50% saturation. In the study a panel of 44 lectins of known carbohydrate specificity was used immobilized onto nitrocellulose membrane. The panel was incu­bated with sIgА preparations and lectin-antibody complexes were detected by protein-A conjugated with horseradish peroxidase. Western-blot analysis was applied to cha­racterize glycosylated polypeptides of sIgA isolated from colostrum of parturient women. sIgA from the colostrum of clinically healthy parturient women contained mainly carbohydrate moieties consisting of N-acetyl-D-glucosamine, sialic acid and D-mannose. Substantial differences in interaction of carbohydrate determinants of slgA isolated from colostrum of individual parturient women with the lectines were found. It is proposed that such characteristics can define sensitivity of mother and child to the action of pathogenic microflora

Level of physiological form of prion in brain and spleen and anti-prpc – antibodies in blood secum of mice is dependent of their ration

It was found that long term feeding of mice with ration supplemented with cattle brain and meat-bone meal (1:1) resulted in producing cellular form of prion in organs of prion-replicated system (brain and spleen) and anti-PrPc antibodies in blood serum. The effect of ration on potential development of transmissible spongiform encepha­lopathies is discussed

Sialylation of anti-histone antibodies in blood serum of systemic lupus erythematosus patients

Anti-histone autoantibodies belong to main marker antibodies used in the diagnostics of systemic lupus erythematosus (SLE) patients. During autoimmune diseases, especially in SLE, prevalent process of inflammation is caused by high titers of auto-antibodies. It is known that the level of IgG sialylation affect their anti-inflammatory properties. The aim of the experiments was to investigate the level of sialylation of anti-histone IgG-antibodies from blood serum of patients with SLE. Investigation was carried out by two methodological approaches. In the first case, the antibodies were purified from blood serum by affine chromatography on histone-Sepharose and protein G-Sepharose followed by Western blot detection of sialic acid residues in the carbohydrate chaines of IgG molecules using sialospecific previously biotinylated Sambucus nigra lectin (SNA). In the second case, IgG-antibodies were isolated from blood serum by affine chromato­graphy on protein G-Sepharose, this fraction was separated on sialylated and not sialy­lated IgG by SNA-Sepharose chromatography followed by checking their affinity to histones by ELISA. In this manuscript, it is shown that the level of sialylation of anti-histone IgG is lower than in control preparation of antibodies. Our data showed that anti-histone autoantibodies are devoid of sialic acid residues at the ends of the carbohydrate chains of Fc-fragments promote their inflammatory properties in patients with SLE

Subcellular localization of adapter protein Ruk, in HEK293 cells

Subcellular localization of adapter protein Ruk1 has been investigated in human embryonic kidney HEK293 cells transfected with pRc/CMV2/Ruk1-Glu-tag. Using immunofluorescence microscopy, it was shown that the recombinant protein was distributed diffusely in both cytoplasm and nucleus, but with punctated structures in the nucleus, which correspond in all probability to the nucleolus. The Ruk1 localization in the nucleus was confirmed by Western blotting of nucleic extracts prepared from transfected HEK293 cells using monoclonal anti-Glu-tag antibodies, as well as from nontransfected HEK293 and U937 cells using polyclonal anti-Ruk antibodies. The ability of affine purified Ruk1 Glu-tagged preparation to bind to DNA from calf thymus, but not to Escherichia coli DNA, was revealed by the immunodot-blot analysis. The nuclear localization of Ruk1 suggests that this adapter protein is involved in some new, yet unrecognized functions, in the eukaryotic cell nucleus