Therapeutic Efficacy and Hepatotoxicity of a Composition of Isoniazid and Dialdehyde Dextran Obtained by Radiochemical Oxidation of Dextran

Tuberculous granulomas were found in all parenchymal organs of mice infected with mycobacteria of BCG vaccine. The number and size of hepatic granulomas decreased, while the count of degenerated and necrobiotic hepatocytes in infected animals increased 3 months after the start of therapy with a composition of isoniazid and dialdehyde dextran. The composition of isoniazid and dialdehyde dextran obtained by radiochemical oxidation of dextran had greater therapeutic efficacy and lower hepatotoxicity. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Suppl. 1, pp. 73–75, 2008     

Effect of Dialdehyde Dextran on Structural Changes in the Liver and Lungs during Chronic BCG Granulomatosis

Male BALB/c mice were intraperitoneally infected with BCG vaccine. Cytomorphological changes in BCG granulomas of the liver and lungs were compared during spontaneous tuberculous inflammation and after intraperitoneal injection of dialdehyde dextran for 5 months. Administration of dialdehyde dextran to mice infected with mycobacteria of BCG vaccine was followed by a decrease in the number and size of BCG granulomas in organs, contributed to the increase in the count of fibroblasts in hepatic and pulmonary granulomas, decreased the severity of destructive changes in the liver parenchyma, promoted reparative processes in hepatocytes, and reduced the degree of fibrosis in the liver and lungs due to a decrease in fibroplastic activity of fibroblasts in BCG granulomas. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Suppl. 1, pp. 113–115, 2008     

Characterization of surface adhesins of lactobacilli used in production of probiotic preparations

We examined production of the protein-lipoteichoic complex detectable by concanavalin A in six lactobacillus strains. A correlation was found between detection of this complex by both reaction with concanavalin A and agglutination with strain-specific antiserum. The cultures were characterized by expression of different types of adhesins. Among them, strains were differentiated with low adhesion activity and intensive production of the protein-lipoteichoic complex and strains with wide adhesin spectrum not producing the complex. We assume that a combination of lactobacillus variants differing in production of the protein-lipoteichoic complex and adhesion potency can be a basis of efficient probiotic preparation. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 141, No. 6, pp. 664–667, June, 2005     

Peculiarities ofin vitro immune response to mycobacterial antigens in inbred I/St mice

Comparative study on inbred mouse stains showed thatin vitro response of T lymphocytes from tuberculosis-susceptible I/St mice to mycobacterial antigens did not differ from that of T lymphocytes from resistant mouse strains. The defect appeared only in the presence of virulent mycobacteria and was not related to T lymphocytes. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 129, No. 6, pp. 661–663, June, 2000     

Stromal cell strains derived from suspension fraction of bone marrow cultures of tumor necrosis factor-deficient mice

Culturing of suspension fraction of a long-term bone marrow culture derived from tumor necrosis factor (TNF)-deficient mice for 75 days produced cells forming adherent cell strains. The cells of all strains expressed RNA of various stromal differentiating markers in various combinations. The cells of many strains simultaneously expressed genes encoding products characteristic of different differentiation lineages. The derived strains maintained hemopoiesis for 10 weeks. RNA-analysis of gene expression by cells of these strains showed that they express a set of various growth factors and cytokines. It was hypothesized that suspension fraction of long-term bone marrow culture derived from TNF-deficient mice includes immature stromal precursor cells, which were never detected in long-term bone marrow culture derived from wild-type mice. Translated fromByulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 138, No. 9, pp. 284–288, September, 2004     

Effect of epithalone on the age-specific changes in the time course of lipid peroxidation inDrosophila melanogaster

The effect of epithalone on the age-specific time course of lipid peroxidation was studied in inadaptiveDrosophila melanogaster strains. A single dose of epithalone at the larval stage decreased the level of conjugated hydroperoxides and Schiff bases throughout the life span of imago. Strain- and sex-specific differences in the time course of the studied sign are characterized. Translated fromByulleten’s Eksperimental’noi Biologii i Meditsiny, Vol. 130, No. 11, pp. 585–588, November, 2000     

Detection of mutations in Mycobacterium tuberculosis genome determining resistance to fluoroquinolones by hybridization on biological microchips

We developed a method of identification of Mycobacterium tuberculosis with simultaneous evaluation of the sensitivity to fluoroquinolones on a biological microchip array. The method of multiplex two-staged PCR followed by hybridization of a biochip makes it possible to detect 8 mutant variants of gyrA gene occurring in fluoroquinolone-resistant strains (∼85% all resistant forms) within 1 day. Using this method we analyzed 107 cultures isolated from patients with tuberculosis and 78 sputum samples. Mutations in gyrA gene were detected in 48 (92%) resistant strains. Natural S95T polymorphism in gyrA gene was detected in all resistant and in 76% sensitive strains. The sensitivity and specificity of the proposed method calculated on the basis of the analysis of sputum samples (n=78) were 94 and 100%, respectively. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 1, pp. 115–120, January, 2008     

Immune response in mice of a new strain ASC (antidepressants sensitive catalepsy)

Inherited predisposition of ASC mice with depressive behavior to catalepsy was accompanied by a significant decrease in the immune response to sheep erythrocytes (compared to parent strains CBA and AKR). The degree of immunosuppression was highest on day 5 after immunization. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 144, No. 8, pp. 188–190, August, 2007     

Effect of hyperlipidemia on thymocyte sensitivity to apoptosis in CBA and C57B1/6 mice

Effects of experimental hyperlipidemia on apoptosis and proliferation of thymocytes in response to mitogens were studied in CBA and C57B1/6 mice. The concentrations of cholesterol in the serum and thymocyte membranes increased in both mouse strains. Spontaneous and dexamethasone-induced apoptosisin vitro and the proliferative response to phytohemag-glutinin and concanavalin A were enhanced in thymocytes from C57B1/6 mice and suppressed in cells from CBA mice. These data suggest opposite reactions of thymocyte to increased serum cholesterol concentration in these two strains, associated with stimulation and suppression of cell activity. Translated fromByulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 130, No. 8, pp. 200–202, August, 2000     

Analysis of the contribution of molecular mechanisms into formation of gonoccocal resistance to tetracycline

We applied complex genetic analysis for evaluation of tetracycline-resistance markers in 129 clinical strains of Neisseria gonorrhoeae from Central, Privolzhskii, and Siberian regions. For detection of mutations in rpsJ gene and MtrRCDE locus we first used minisequence reaction followed by identification of products by MALDI-TOF mass spectrometry. The incidence of detection of resistance markers among the analyzed strains were: tetM — 3.1%, mutations in genes rpsJ — 82.2 %, penB — 62.8%, and mtrR — 54.3%. The analyzed genetic markers were not detected in 17.5% strains. tetM gene was detected in only 12.5% strains from the Central Region. No differences were revealed in regional distribution of other genotypes. Genotypes tetM _pres, rpsJ _mut, mtrR _mut, and rpsJ _mut, penB _mut, mtrR _mut reliably predict tetracycline resistance. Microbiological and genetic testing of tetracycline resistance yielded similar results. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Supplement 2, pp. 61–66, April, 2007     

Evaluation of a Fluorescence Hybridisation Assay Using Peptide Nucleic Acid Probes for Identification and Differentiation of Tuberculous and Non-Tuberculous Mycobacteria in Liquid Cultures

 The performance was evaluated of a fluorescence in situ hybridisation assay using peptide nucleic acid probes (Dako Probe MTB Culture Confirmation Test; Dako, Denmark) for identification of Mycobacterium tuberculosis complex (MTC) organisms and differentiation between tuberculous and non-tuberculous mycobacteria (NTM) in material taken directly from Bactec 12B (Becton Dickinson, USA) and MB/BacT (Organon Teknika, USA) bottles. The test was applied to 129 smear-positive (Ziehl-Neelsen stain) clinical specimens, 48 previously identified clinical strains of mycobacteria (12 MTC and 36 NTM), and 51 reference strains (7 MTC and 44 NTM) which were all previously inoculated into Bactec 12B and MB/BacT bottles. The sensitivity and specificity of the assay for MTC-positive cultures was 87.6% and 100%, respectively, for Bactec 12B, and 100%, respectively, for MB/BacT. The sensitivity and specificity of the assay for NTM-positive cultures was 100% for both media.     

Role of Rac and Rho-GDI Alpha in the Frequency-dependent Expression of h1-calponin in Vascular Smooth Muscle Cells under Cyclic Mechanical Strain

Phenotype transformation of vascular smooth muscle cells (VSMCs) has been reported to be directly influenced by the frequency of mechanical strain. This study explored the effects of different frequencies of mechanical strain on expression of phenotype marker h1-calponin and the possible mechanism. VSMCs were subjected to cyclic strains of 10% elongation at 1 and 2 Hz for 24 h by using a Flexercell strain unit. The protein expression of h1-calponin was assessed by Western blotting and the possible protein kinases involved were evaluated by their specific inhibitor or targeted siRNA ‘knock-down.’ The results showed that cyclic strains modulated the expressions of h1-calponin, phospho-p38, Rac and Rho-guanine nucleotide dissociation inhibitor alpha (Rho-GDIα) in nonlinear frequency-dependent manners. This nonlinear frequency-dependent change of h1-calponin expression could be blocked by a specific p38 inhibitor, SB202190. The changed expression of phospho-p38 induced by the frequencies of cyclic strain was reversed by targeted siRNA ‘knock-down’ of Rac, while enhanced by targeted siRNA ‘knock-down’ of Rho-GDIα. These results suggest that the frequency-dependent expression of h1-calponin under cyclic strain is mediated at least partly by the regulation of Rac and Rho-GDIα expression on the activation of p38 pathway. Ming-Juan Qu and Bo Liu contributed equally to this work.     

Epitopic mapping of different mycobacteria with the use of monoclonal antibodies

Monoclonal antibodies reacting with different mycobacteria are prepared. They are used for epitopic mapping ofMycobacteria tuberculosis H37Rv, M. bovis bovinus 8, M. avium, M. kansasii, andM. smegmatis. On the basis of the epitope maps 129 cultures of mycobacteria were identified by immunoblotting with the antibodies. The results of this identification coincided with those of microbiological investigation in 61.5–80% cases, depending on the genus of mycobacteria. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 125, No. 3, pp. 329–331, March, 1997     

Effect of Thyroxin on Behavior of Mice with Inherited Difference in Predisposition to Catalepsy

We studied effects of chronic thyroxin administration (2 mg/l, for 60 days) on motor activity, anxiety and depression-like behavior in cataleptic (ASC/Icg) and non-cataleptic (AKR/J) strains of mice. No effects of thyroxin on anxiety indicators in “open field” and “light/dark” tests were revealed in mice of the strains under study. At the same time, thyroxin increased moveability in the “open field” test in AKR/J mice and produced an antidepressant effect in the “forced swimming” test in animals from ASC/Icg strain. Obtained results are indicative of the role of inherited predisposition to catalepsy in determining the sensitivity to thyroid hormones. Translated from Byulleten’ Experimental’noi Biologii I Meditsiny, Vol. 147, No. 2, pp. 177-180, February, 2009     

Lymphadenitis in children is caused by <Emphasis Type="Italic">Mycobacterium avium hominissuis</Emphasis> and not related to ‘bird tuberculosis’

Mycobacterium avium is the most commonly encountered mycobacterium species among non-Mycobacterium tuberculosis complex (nontuberculous mycobacteria) isolates worldwide and frequently causes lymphadenitis in children. During a multi-centre study in The Netherlands that was performed to determine the optimal treatment for mycobacterial lymphadenitis, concern was expressed in the media about the possible role of birds as sources of these M. avium infections, referred to as ‘bird tuberculosis.’ To examine the involvement of birds in mycobacterial lymphadenitis, 34 M. avium isolates from lymphadenitis cases were subjected to IS1245 restriction fragment length polymorphism (RFLP) typing. This genotyping method enables the distinction of the subspecies M. avium subsp. hominissuis and the ‘bird-type’ M. avium spp. avium. Highly variable RFLP patterns were found among the lymphadenitis M. avium isolates, and all belonged to the M. avium hominissuis subspecies. A relation to pet birds in the etiology of mycobacterial lymphadenitis could not be established, and the source of the infections may be environmental.     

Nonradioactive single-strand conformation polymorphism analysis for detection of fluoroquinolone resistance in Mycobacteria

A simple, rapid, and nonradioactive method for routine detection of fluoroquinolone resistance in mycobacteria is described. A single-strand conformation polymorphism (SSCP) methodology, based on the use of mini-gels and silver staining of DNA, was optimized for the analysis of denatured DNA products obtained by polymerase chain reaction (PCR) from thegyrA gene involved in fluoroquinolone resistance in mycobacteria. The method was successfully applied to fluoroquinolone-susceptible and -resistant laboratory strains ofMycobacterium smegmatis and to clinical strains ofMycobacterium tuberculosis isolated from patients who developed resistance during the course of fluoroquinolone treatment.     

BCG Granulomatosis in Adult Mice of Different Strains with a History of Intrauterine Hypoxia

Specific features of the formation and spontaneous regression of BCG granulomas, depending on mouse genotype and history of intrauterine hypoxia, were detected in 2-month-old male CBA and C57Bl/6 mice. The numerical density of granulomas, their size and (partially) cell composition varied. There are good grounds to assume that the number of BCG granulomas in the liver, irrespective of the experimental conditions and animal strains, decreases after elimination of M. tuberculosis persisting in granuloma phagocytes and subsequent migration of cells constituting the granuloma. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Suppl. 1, pp. 93–96, 2008     

The prevalence of plasmid-mediated AmpC β-lactamases among clinical isolates of Escherichia coli and Klebsiella pneumoniae from five children’s hospitals in China

The purpose of this study was to investigate the prevalence of plasmid-mediated AmpC β-lactamases in Escherichia coli and Klebsiella pneumoniae from five children’s hospitals in China. A total of 494 E. coli and 637 K. pneumoniae isolates were collected from five children’s hospitals in China from 2005 to 2006. The isolates with decreased susceptibility to cefoxitin were subjected to confirmation test with 3-aminophenyl boronic acid. Polymerase chain reaction (PCR) amplification of the blaAmpC, blaTEM, blaCTXM, and blaSHV genes and their gene sequencing were performed. Transconjugants were achieved by conjugation experiments. Plasmid-mediated AmpC β-lactamases were found in 10.1% of K. pneumoniae (64/637) and in 2.0% of E. coli (10/494) strains. The proportion of plasmid-mediated AmpC-producing strains significantly increased from 2005 (2.6%) to 2006 (9.3%) (p<0.001). The DHA-1-producing isolates were the most prevalent type (93.2%, 69/74). The sequences of blaDHA-1 genes were all identical to those from the GenBank. Strains of blaCMY-2 were isolated from five isolates (6.8%), which were all from E. coli. One sequence of blaCMY-2 differs from blaCMY-2 in the GenBank. Eighteen of the 74 (24.3%) AmpC-producing K. pneumoniae and E. coli isolates coproduced an extended-spectrum β-lactamase (ESBL). Cefoxitin resistance was transferred to 15 of the 74 positive strains (20.3%). Our study has demonstrated the occurrence of plasmid-mediated AmpC β-lactamases in E. coli and K. pneumoniae in Chinese pediatric patients and DHA-1 type AmpC enzymes had the highest prevalent rate. The CMY-2 AmpC β-lactamases from the children’s hospitals in China in this study are the first reported. Hence, continuous surveillance of the prevalence and evolution of AmpC β-lactamase is important.     

Therapeutic Efficacy of a Composition of Amphotericin B and Dialdehyde Dextran in Kidney Damage in Mice of Various Strains with Systemic Candidiasis

The therapeutic efficacy of a composition of amphotericin B and dialdehyde dextran was much higher than that of amphotericin B in the therapy for systemic candidiasis. This conclusion was derived from the earlier and progressive decrease in the numberand size of candidal granulomas in the kidneys. The composition of amphotericin B and dialdehyde dextran was more potent than amphotericin B in decreasing the nephrotoxiceffect of C. albicans. The opposite strains of CBA and C57Bl/6 mice differed in morphologicalsigns of granulomatosis in the kidneys, but not in the nephrotoxic effect of C. albicans metabolites. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 149, No. 12, Suppl. 1, pp. 76–79, December, 2008     

Evidence for limited exchange of avian influenza viruses between seaducks and dabbling ducks at Alaska Peninsula coastal lagoons

Avian influenza virus (AIV) prevalence and sequence data were analyzed for Steller’s eiders (Polysticta stelleri) to assess the role of this species in transporting virus genes between continents and maintaining a regional viral reservoir with sympatric northern pintails (Anas acuta). AIV prevalence was 0.2% at Izembek Lagoon and 3.9% at Nelson Lagoon for Steller’s eiders and 11.2% for northern pintails at Izembek Lagoon. Phylogenetic analysis of 13 AIVs from Steller’s eiders revealed that 4.9% of genes were of Eurasian origin. Seven subtypes were detected, including two also observed in northern pintails. No AIV strains were highly similar (> 99%) at all gene segments between species; however, highly similar individual genes were detected. The proportion of highly similar genes was greater within rather than between species. Steller’s eiders likely transport AIV genes between continents through long-distance migratory movements. Differences in AIV prevalence, subtype distribution, and the proportion of highly similar genes suggest limited AIV exchange between Steller’s eiders and northern pintails at Alaska Peninsula coastal lagoons during autumn.