Corynebacterium spp.: relationship of pathogenic properties and antimicrobial resistance.

Corynebacterium spp. are part of the human microbiome, but can cause the development of inflammatory diseases of various localization. Purpose - to evaluate the relationship between pathogenic properties and resistance to antimicrobial drugs (AMD) of Corynebacterium spp. from patients with inflammatory diseases of the respiratory tract. Strains of Corynebacterium spp. isolated from patients with inflammatory diseases of the respiratory tract (99 pcs.) and practically healthy individuals (33 pcs.). Isolates were identified by mass spectrometric method (MALDI-ToFMS), their adhesive and invasive activity on Hep-2 cells, cytopathic effect (CPE) in CHO-K1 cell culture, and resistance to antimicrobial drugs (AMD) were determined. Indicators of adhesion (3.65±0.679(CFU±m)x102/ml), invasion (1.72±0.230 (CFU±m)x102/ml), cytotoxicity (69.1±3.8% of dead CHO-K1 cells ) Corynebasterium spp. strains isolated from patients are higher (p≤0.05) than similar indicators in practically healthy people. 90.9% of isolates from patients had resistance to AMD, in most cases (57.6±4.9%) resistance to only one AMP was noted, less often to two (25.2±4.3%), three or more (8.08±2.7%). According to the results of correlation-regression analysis, pathogenic properties (adhesiveness, invasiveness, cytotoxicity) of Corynebacterium spp. strains isolated from patients are in close direct relationship with resistance to AMD. This indicates the importance of identifying strains of non-diphtheria corynebacteria resistant to AMDs, which, under the influence of developing resistance to AMDs, can increase their pathogenic potential, moving from commensalism to parasitism.

AnFiSA: An open-source computational platform for the analysis of sequencing data for rare genetic disease.

Despite genomic sequencing rapidly transforming from being a bench-side tool to a routine procedure in a hospital, there is a noticeable lack of genomic analysis software that supports both clinical and research workflows as well as crowdsourcing. Furthermore, most existing software packages are not forward-compatible in regards to supporting ever-changing diagnostic rules adopted by the genetics community. Regular updates of genomics databases pose challenges for reproducible and traceable automated genetic diagnostics tools. Lastly, most of the software tools score low on explainability amongst clinicians. We have created a fully open-source variant curation tool, AnFiSA, with the intention to invite and accept contributions from clinicians, researchers, and professional software developers. The design of AnFiSA addresses the aforementioned issues via the following architectural principles: using a multidimensional database management system (DBMS) for genomic data to address reproducibility, curated decision trees adaptable to changing clinical rules, and a crowdsourcing-friendly interface to address difficult-to-diagnose cases. We discuss how we have chosen our technology stack and describe the design and implementation of the software. Finally, we show in detail how selected workflows can be implemented using the current version of AnFiSA by a medical geneticist.

[The immune-enzyme techniques of analysis in diagnostic of cholera.]

The optimal conditions of arrangement of direct alternative of flatbed enzyme-linked immunosorbent assay and dot-immunoanalysis with application of monoclonal peroxidase conjugates for express identification of comma bacillus of serogroups O1 and O139 both in hospital and field conditions without device support. The direct technique enzyme-linked immunosorbent assay in flatbed alternative shortens time of analysis up to 70-80 minutes and in case of dot enzyme-linked immunosorbent assay on membrane - up to 70-90 minutes. It is established that in case of analysis in conditions of room temperature (20-25 oC) sensitivity of techniques remains at initial level.

[Evaluation of toxin producing abilities of non-O1/non-O139 serogroup Vibrio cholerae isolated from humans].

AIM: Determination of non-O1/non-O139 Vibrio cholerae toxin (CT) gene expression by using EIA, and biological effect of non-O1/non-O139 V. cholerae supernatant on cell cultures evaluation. MATERIALS AND METHODS: 39 V. cholerae strains from various serological groups were studied. Hemolytic activity of strains was determined by using Greig test, and cholera toxin production--in GM1-EIA and in continuous cell lines by registering cytotonic, cytotoxic and proteolitic effect. RESULTS: GM1-EIA method does not detect CT production in 29 museum strains of non-O1/non-O139 V. cholerae in vitro. CT was detected only in 1 non-O1/non-O139 V. cholerae strain supernatant with OD = 0.577 that is substantially lower than in O1 V. cholerae strains (OD = 2.176). In cell cultures non-O1/non-O139 V. cholerae supernatants diluted to 1:100 caused elongation only in single cells. CONCLUSION: Cytological model is a more sensitive technique to evaluate toxin producing abilities of non-O1/non-O139 V. cholerae strains and is appropriate for use.

[Identification of Vibrio cholerae serogroups O1 and O139 by monoclonal antibodies in the slide agglutination test].

The agglutinating properties of MCA-O1 of the IgG class and MCA-O139 of the IgM class towards epitopes of O-antigen of Vibrio cholerae O1 and accordingly Vibrio cholerae O139 were studied. The ascitic and cultural fluids by hybridomas F8G12 and D11 deposited in the specialized Collection of Cell Cultures of Vertebrates (Saint Petersburg) under RKKK (II) 386 D and RKKK (II) 674 D were the sources of monoclonal immunoglobulins. The advantage of diagnostic monoclonal immunoglobulins is that they are distinguished for strict specificity and their use in practical health care contributes to the higher specificity of a laboratory test for cholera and to its shorter performance.

Morphofunctional changes in neurons of hippocampal fields and neocortical layers in rats treated with Polydan.

Light microscopy revealed four types of neurons differing by staining intensity and size in each of the pyramidal neuron populations in the dorsal hippocampal fields and layer V of the neocortical somatosensory area and in granular cells of the dentate fascia and stellate cells of layer II of the same cerebrocortical area in rats. Treatment with Polydan according to different protocols led to redistribution of these types of cells, which attests to stimulation of synthetic processes in these brain areas, rather than to similar sequence of the involvement of different brain areas in this process. Polydan promoted the increase in the mean number of nucleoli in the nuclei of neurons of all types, the degree of this increase being different for each type. It seems that morphological signs (staining intensity and number and size of nucleoli in the nuclei) reflect certain functional states of the neurons in the homogeneous populations. Presumably, various factors can stimulate transition of neurons from one morphofunctional status into another.

[Some aspects of occupational hygiene in females engaged into nickel production in Murmansk region]. / Nekotorye aspekty gigieny truda zhenshchin, zaniatykh v proizvodstve nikelia na predpriiatiiakh Murmanskoi oblasti.

The authors present hygienic evaluation of work conditions for women engaged into nickel production, when related to physiologic features of female organism.

[Effect of polidan on the conditioned memory and structural and metabolic parameters of the rat brain neurons]. / Polidan--vliianie na uslovno-reflektornuiu pamiat' i strukturno-metabolicheskie pokazateli v neironakh mozga krys.

New experimental data on the mnemotropic (nootropic) properties of the well-known drug polidan (a DNA derivative) are summarized. The results obtained by histological techniques showed changes in the brain structure, indicating protein synthesis activation under the action of polidan. These results confirm prognostic function of the concept of evolutionary-molecular basis of memory.

The major messenger ribonucleoprotein particle protein p50 (YB-1) promotes nucleic acid strand annealing.

p50, a member of the Y-box binding transcription factor family, is tightly associated with eukaryotic mRNAs and is responsible for general translational regulation. Here we show that p50, in addition to its previously described ability to melt mRNA secondary structure, is capable of promoting rapid annealing of complementary nucleic acid strands. p50 accelerates annealing of RNA and DNA duplexes up to 1500-fold within a wide range of salt concentrations and temperatures. Phosphorylation of p50 selectively inhibits DNA annealing. Moreover, p50 catalyzes strand exchange between double-stranded and single-stranded RNAs yielding a product bearing a more extended double-stranded structure. Strikingly, p50 displays both RNA-melting and -annealing activities in a dose-dependent manner; a relatively low amount of p50 promotes formation of RNA duplexes, whereas an excess of p50 causes unwinding of double-stranded forms. Our results suggest that the alteration of nucleic acid conformation is a basic mechanism of the p50-dependent regulation of gene expression.

The major mRNA-associated protein YB-1 is a potent 5' cap-dependent mRNA stabilizer.

mRNA silencing and storage play an important role in gene expression under diverse circumstances, such as throughout early metazoan development and in response to many types of environmental stress. Here we demonstrate that the major mRNA-associated protein YB-1, also termed p50, is a potent cap-dependent mRNA stabilizer. YB-1 addition or overexpression dramatically increases mRNA stability in vitro and in vivo, whereas YB-1 depletion results in accelerated mRNA decay. The cold shock domain of YB-1 is responsible for the mRNA stabilizing activity, and a blocked mRNA 5' end is required for YB-1-mediated stabilization. Significantly, exogenously added YB-1 destabilizes the interaction of the cap binding protein, eIF4E, with the mRNA cap structure. Conversely, sequestration of eIF4E from the cap increases the association of endogenous YB-1 with mRNA at or near the cap, and significantly enhances mRNA stability. These data support a model whereby down-regulation of eIF4E activity or increasing the YB-1 mRNA binding activity or concentration in cells activates a general default pathway for mRNA stabilization.

[Major mRNP proteins in the structural organization and function of mRNA in eukaryotic cells]. / Mazhornye belki mRNP v strukturnoi organizatsii i funktsionirovanii mRNK v kletkakh éukariot.

The properties of two universal major proteins of cytoplasmic mRNP, p50 and the poly(A)-binding protein (PABP), are summarized. Their roles in formation of polyribosomal and free inactive mRNP are considered, with the focus on the authors' studies of p50. The parts these mRNP proteins play in translation regulation, stability, and localization of mRNA are described, and the the possible mechanisms of their function are discussed.

[Sex ratio in early embryonal mortality in man]. / K voprosu o sootnoshenii polov pri rannei émbrional'noi letal'nosti u cheloveka.

The problem of the functioning specificity of sex chromosomes during the early stages of embryogenesis in man and the associated problem of the sex ratio in spontaneous and induced abortions, as well as in newborns, remains open. We have conducted a cytogenetic examination of 342 spontaneous abortions divided into three clinical groups on the basis of the severity of the developmental disturbances of the embryo: spontaneous abortions sensu stricto with a developed embryo without any significant intrauterine delay of development (n = 100), nondeveloping pregnancies (n = 176), and anembryonic fetuses (n = 66). The frequency of chromosomal mutations in these groups was 22.0, 48.3, and 48.5%, respectively. Statistical analysis has demonstrated significant differences between the studied groups in the frequencies of the normal and abnormal karyotypes: the major contributions to these differences were associated with autosomal trisomy, triploidy, and 46,XY karyotype. The presence of 46,XY may reflect specific genetic mechanisms of prenatal mortality of embryos with normal karyotype, associated with sex and/or with the imprinting of X-chromosomes. The sex ratio in spontaneous abortions with normal karyotype was as follows: 0.77 for spontaneous abortions with well-developed embryos without any significant intrauterine delay of development; 0.60 for non-developing pregnancies; and 0.31 for anembryonic fetuses. An analysis of DNA from the embryos and their parents has demonstrated a low probability of contamination of cell cultures with mother cells as a possible source of prevalence of embryos with 46,XX karyotype among spontaneous abortions. Nondeveloping pregnancies and anembryonic fetuses showed statistically significant differences in the sex ratio (1.11) from the control group consisting of medical abortions. Differences in the sex ratio were due to an increasingly lower proportion of embryos with karyotype 46,XY (relative to the expected one) among the fetuses with an increased severity of developmental disturbances. The statistical "chances ratio" index also provided evidence that embryos with 46,XY karyotype had a higher propensity to produce a well-formed fetus as compared with the female embryos. We propose that the expression of genes of the maternal X-chromosome in XY embryos supports a more stable development during early embryogenesis as compared with XX embryos. In the latter case, normal development is coupled with the operation of an additional mechanism for compensation of the dose of X-linked genes. Operation of this mechanism increases the probability of disturbances in female embryos. A higher viability of XY embryos during the early stages of ontogenesis in man appears to explain their underrepresentation in samples of spontaneously aborted embryos and appears to be the major factor responsible for the deviation of the sex ratio from the theoretically expected value.

[Polymorphism of trinucleotide CAG-repeats of the first exon of the androgen receptor gene in the Tomsk population]. / Polimorfizm trinukleotidnykh CAG-povtorov pervogo ékzona gena retseptora androgenov v populiatsii g. Tomska.

Polymorphism of the CAG repeat of exon 1 of the androgen receptor (AR) gene was analyzed in the Tomsk population. In total, 12 alleles varying in size from 285 to 318 bp (21-32 CAG units) were revealed. The allele frequency distribution did not differ from the normal one. No difference in allele frequencies was detected between men and women of the same generation. The observed heterozygosity was equal to the expected one (0.88 +/- 0.03). Compared with other populations, the Tomsk population displayed a narrower allele spectrum and a bias of the most common allele to a greater repeat number. The results obtained may reflect specific population genetic processes characteristic of young developing populations.

[The absence of uniparental X-chromosome inheritance in spontaneous abortuses with a 46,XX karyotype]. / Otsutstvie odnoroditel'skogo nasledovaniia X-khromosom u spontannykh abortusov s 46.XX-kariotipom.

The problem of the presence of imprinted regions on the X-chromosome and the possible influence of the imprinted expression of X-linked genes on the embryonic development in man remains largely unsolved. A comparison of the uniparental inheritance of chromosomes or of their regions having different phenotypic manifestations provides an instrument with which to study the phenomenon of genomic imprinting at the chromosomal level. Assuming that the imprinted inactivation of X-chromosomes is functionally significant for embryonic development, we have studied several polymorphic micro- and minisatellite loci of X-chromosomes in 52 fetuses with karyotype 46,XX, which were spontaneously aborted during the first trimester of pregnancy. The purpose was to determine the contribution of uniparental disomy for the X-chromosome in any disturbances of the embryonic development. We found that inheritance of X-chromosomes was biparental in the studied embryos, suggesting the absence of any significant contribution of the parental origin of the X-chromosome to embryonic mortality occurring between 4 and 12 weeks of development.

Interaction of the universal mRNA-binding protein, p50, with actin: a possible link between mRNA and microfilaments.

We have shown previously that p50 is the most abundant protein associated with a variety of eukaryotic mRNAs and exhibits about 98% amino acid sequence identity to mammalian Y-box binding transcription factors. The dual function of p50 in the cell as a regulator of both transcription and translation has been suggested. To gain insight into the role of p50 in these processes, we performed the yeast two-hybrid screen to identify p50 molecular partners. Here we report the identification of actin as a p50-interacting protein. Coimmunoprecipitation of p50 and actin from HeLa extracts as well as in vitro binding studies indicate specificity and a high affinity for the interaction between p50 and actin. Interestingly, p50 binding to actin is affected by mRNA; binding was observed at a low p50/mRNA ratio and was greatly reduced at higher ratios. Since the p50/mRNA ratio appears to be important for mRNA translatability, we speculate that p50 can regulate the attachment of mRNA to the actin network depending on its translational activity. Using immunofluorescence, we show that p50 binds to actin filaments in permeabilized cells and causes actin fibers to bundle in vitro. Together, these findings support the view that p50 may play an important role in mRNA transport, anchoring, and localization on actin filaments in the cell.

Translational regulation by Y-box transcription factor: involvement of the major mRNA-associated protein, p50.

p50, the major core protein of messenger ribonucleoprotein particles (mRNPs), is a universal protein found exclusively in association with different mRNA species in the cytoplasm of somatic mammalian cells. Furthermore, p50 is the most abundant and tightly bound protein within both inactive mRNPs and active mRNPs derived from polysomes, although the latter contain a lower level of p50. Recent experiments have revealed that, depending on the p50 to mRNA ratio, p50 may either act as a repressor or an activator of protein synthesis. On the other hand, p50 exhibits about 98% amino acid sequence identity to mammalian transcription factors that bind specifically to Y-box containing DNA. Thus, it is a counterpart of the Y-box binding proteins which are found in bacteria, plants and animals, exhibiting multiple biological activities ranging from transcriptional regulation of a wide variety of genes to 'masking' mRNA activity in germinal cells. This review summarizes our current knowledge of p50 structure and function. It also discusses the biological roles of p50 and related proteins in gene expression and describes the likely mechanisms of their action.

The major core protein of messenger ribonucleoprotein particles (p50) promotes initiation of protein biosynthesis in vitro.

The major core protein of cytoplasmic messenger ribonucleoprotein particles (p50) has been shown previously to inhibit protein synthesis in vitro and in vivo. Furthermore, p50 is highly homologous to the Y-box-binding transcription factor family of proteins, binds DNA containing the Y-box motif, and thus may have a dual function in cells as a regulator of both transcription and translation. Here we show that binding or removal of p50 from rabbit reticulocyte lysate by monospecific antibodies to p50 strongly inhibits translation of endogenous and exogenous globin mRNAs as well as prokaryotic beta-galactosidase mRNA in a rabbit reticulocyte cell-free system. Thus, depending on the conditions, p50 not only may act as a translational repressor, but may also be required for protein synthesis. Translation inhibition with anti-p50 antibodies is not a result of mRNA degradation or its functional inactivation. The inhibition does not change the ribosome transit time, and therefore, it does not affect elongation/termination of polypeptide chains. The inhibition with anti-p50 antibodies is followed by a decay of polysomes and accumulation of the 48 S preinitiation complex. These results suggest that p50 participates in initiation of protein biosynthesis. Although uninvolved in the formation of the 48 S preinitiation complex, p50 is necessary either for attachment of the 60 S ribosomal subunit or for previous 5'-untranslated region scanning by the 43 S preinitiation complex.

Overexpression in COS cells of p50, the major core protein associated with mRNA, results in translation inhibition.

p50, the major core protein of messenger ribonucleoprotein particles (mRNPs) in the cytoplasm of somatic mammalian cells, has been characterized previously as a member of the Y-box binding transcription factor family of proteins (YB-protein) by both high structural homology and ability to bind specifically the Y-box sequence in double-stranded DNA. YB proteins are present in a whole range of cell types and some have been identified as germ-specific cytoplasmic proteins masking stored mRNA from translation. Western blot analysis of the distribution of p50 in subcellular fractions of COS-1 cells shows that p50 is a cytoplasmic protein quantitatively associated with mRNA, both in polyribosomes and in free mRNPs. The level of p50 in COS-1 cells determined by Western immunoblotting is 0.10% of total protein, which is nearly equimolar to that of ribosomes and is approximately 5-10-fold higher than the mRNA level. Transient transfection of COS-1 cells with a p50-expressing vector results in a dramatic inhibition of protein synthesis. A control transfection with a vector expressing a frameshift mutant of p50 does not cause translation inhibition. Therefore the increase in p50 protein level is responsible for the inhibitory effect in these cells.

[X-chromosome inactivation in mammalian ontogeny]. / Inaktivatsiia X-khromosomy v ontogeneze mlekopitaiushchikh.

This review is dedicated to a very interesting problem of developmental genetics: dose compensation of the X-chromosome genes in somatic cells of female mammals. A sequence of events related to X-chromosome inactivation during cell differentiation at the early embryonic stages, association of distributed X-chromosome heterochromatization with developmental pathologies, evolutionary origin of X-inactivation, and its relationship with genomic imprinting are considered.

[The major cytoplasmic mRNP protein, p50, is required for efficient mRNA translation in vitro]. / Mazhornyi belok tsitoplazmaticheskikh mRNP, p50, neobkhodim dlia éffektivnoi transliatsii mRNK in vitro.

The major cytoplasmic mRNP protein of somatic cells, p50, is the member of the Y-box-binding transcription factor family and can control gene expression at two levels including mRNA transcription and translation. It has been demonstrated that p50 is responsible for the inactive state of mRNA within free mRNPs. In this work, we show that the Y-box-containing DNA (Y-DNA) predominantly binds to p50 in rabbit reticulocyte lysates and causes translation inhibition of the endogenous and exogenous globin mRNA and prokaryotic beta-galactosidase mRNA. Preincubation of Y-DNA with purified p50 prevents the inhibition. Inhibition of protein biosynthesis by the Y-DNA is not due to the degradation or functional inactivation of mRNA. The inhibition is associated with the decay of polyribosomes and dissociation of a newly synthesized protein from the ribosomes. The data indicate that Y-DNA inhibits protein biosynthesis predominantly at the initiation stage and that p50 is an essential component of the translation initiation apparatus.