E M Malkova

State Research Center of Virology and Biotechnology VECTOR, Novo-Nikolaevsk, Novosibirsk, Russia

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Publications (25)1.27 Total impact

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    ABSTRACT: The synergistic action of the cytostatic drug cyclophosphamide (CP) and fragmented exogenous DNA causes illness and death in mice (Dolgova et al., 2011–2013). The observed “delayed death” effect was most clearly pronounced when the DNA preparation was administered 18 to 30 h after CP treatment. This time span was termed the “death window.”It was found that injections of exogenous DNA result in a sustained increase in bone-marrow cell (BMC) apoptosis, which occurs throughout the time of DNA administration (18–30 h). Exogenous DNA, both allogeneic and belonging to various taxa, induces BMC apoptosis. Plasmid DNA has the greatest effect on apoptosis induction. The analysis of reduction and restoration of the BMC subpopulations as the mice progressed to death revealed a virtually complete loss of the 12–20-μm fraction of the cell population (about 3–4% vs. 35–40% in the control), which corresponds to the maximum leukopenia on day 3 after CP treatment. However, the relative number of CD34+ hematopoietic stem cells (HSCs) from day 15 and until the end of the observation constituted 1.2–1.4%, which corresponds to the wild-type range. Comparison of BMC smears from the sternal bone marrow of the CP and CP + DNA groups of mice indicated that the BMC populations isolated from CP + DNA animals lack young committed lymphopoiesis progenitor cells. Moreover, the affected mice had immature blast cell types in their blood, which was never observed in healthy or CP-treated mice. Pathological and morphological analyses show that starting from posttreatment day 9, mice that received CP + DNA preparations displayed pronounced morphological changes in their lungs, liver, pancreas, central and peripheral immune system organs, and brain. Most of the pathological changes observed are consistent with a severe inflammatory response. This suggestion was proven by structural equivalents of functional involution of lymphoid organs, such as the thymus, spleen, and lymph nodes.We speculate that the death of treated animals resulted from multiple organ dysfunctions caused by accidental involution of lymphoid organs and the systemic inflammatory response syndrome, both associated with injections of fragmented exogenous DNA into experimental animals within the “death window,” which corresponds to the final step in the repair of the majority of CP-induced double-strand breaks.
    Russian Journal of Genetics: Applied Research. 01/2013; 3(4).
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    ABSTRACT: To study efficacy of anaferon pediatric in mice infected by pandemic influenza virus A(H1N1/09)v. Influenza virus strain A/California/07/2009 (H1N1)v was used. Three groups of BALB/c mice intranasally inoculated with influenza virus were studied. First group received solution of Anaferon pediatric during 5 days before and 8 days after inoculation, 2nd group received Tamiflu during 5 days after inoculation. Distilled water was administered orally to mice from control group. It was shown that Anaferon pediatric used as preventive and treatment agent in mice intranasally inoculated with 100% infectious dose of influenza virus strain A/ California/07/2009 (H1N1)v had antiviral effect, which expressed in 10-fold decreased reproduction of influenza virus in lungs of infected mice compared to control group measured 4, 6, and 8 days after inoculation. Use of anaferon pediatric before and after inoculation with influenza virus A(H1N1/09)v was not less effective than use of Tamiflu after inoculation.
    Zhurnal mikrobiologii, epidemiologii, i immunobiologii 01/2011;
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    ABSTRACT: Molecular epidemiological study of novel strain of Rubella virus isolated during the outbreak in Western Siberia in 2004 was described. Detailed phylogenetic analysis performed based upon entire SP-region, which encodes all three Rubella structural proteins (C, E2, and E1), was implemented. This analysis provides characterization of this strain and classifies it as 1H genotype, thereby correcting previous classification of this strain based upon shorter nucleotide sequence, only encoding E1 protein. Therefore, this study identified the genotype of the Rubella virus not previously detected in Western Siberia (and even entire Russian Federation), which highlights the importance of more extensive characterization of genetic variability of the Rubella virus, especially with regard to potential influence of vaccination on the Rubella virus mutagenesis.
    Molekuliarnaia genetika, mikrobiologiia i virusologiia 01/2011;
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    ABSTRACT: The study of basic biological properties of H5N1 subtype strain isolated during an outbreak among wild birds in Russia in 2010 was presented. The study was carried out using conventional methods according to the WHO recommendations. H5N1 influenza virus isolated in Siberia belonged to clade 2.3.2 of the hemagglutinin gene; the phylogenetic analysis was performed. The antigenic characteristics and the basic genetic markers of biological properties were studied. It was shown that all strains were highly pathogenic for chickens and white mice. Thus, it was shown that in Russia in the 2010 H5N1 virus phylogenetically closely related to Asian variants caused epizootic among wild birds. The potential danger of this variant of the virus for humans was confirmed by different methods. We discussed the possibility of formation of H5N1 influenza natural focus.
    Molekuliarnaia genetika, mikrobiologiia i virusologiia 01/2011;
  • L N Shishkina, E M Malkova
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    ABSTRACT: Cells from the rat bronchoalveolar lavage fluid comprising over 70% of neutrophils were labeled with 51Cr and administered intraperitoneally or intravenously to control animals and to rats which were injected subcutaneously with 2 mg of ketalog 5 days before the onset of experiment. Blood samples, lung, hepatic, splenic, renal, small intestinal, and muscular biposies were taken 1 hour after cell infusion. The intraperitoneal administration of 51Cr-labeled neutrophils did not significantly increase the radioactivity count rate in any organ examined. The rise was most notable in the lungs after v/v infusion of labeled neutrophils. Rats given kenalog showed a low level of radioactivity in the lung tissue compared with controls probably because the glucocorticoid caused a specific change in the mechanism of uptake of labeled neutrophils.
    Vestnik Rossiĭskoĭ akademii meditsinskikh nauk / Rossiĭskaia akademiia meditsinskikh nauk 01/2011;
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    ABSTRACT: Anaferon (pediatric formulation) administered in the therapeutic-and-prophylactic regimen to mice receiving intranasally 100% infecting dose of A/California/07/2009(H1N1)v influenza virus exhibited an antiviral effect and 10-fold reduced the production of influenza virus in the lungs of infected mice on days 4, 6, and 8 after infection compared to the control (distilled water). The efficiency of Anaferon (pediatric formulation) administered before and after infection with A/California/07/2009(H1N1)v influenza virus was not inferior to the use of Tamiflu after infection.
    Bulletin of Experimental Biology and Medicine 05/2010; 149(5):612-4. · 0.34 Impact Factor
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    ABSTRACT: To analyze influenza viruses isolated in the 2008-2009 autumn-winter season, and to test sera collected in the south of Western Siberia during the beginning and the end of the epidemic seasons from 2007 until the A/H1N1 pandemic. Total 149 clinical samples were analyzed and 2190 blood sera were tested. During the 2008-2009 season 17 influenza viruses were isolated. 9 of these were A/H1N1, 5-were A/H3N2, and 3 were influenza B viruses. The nucleotide sequences and amino acid composition of influenza A virus hemagglutinin (HA) were compared with reference strains. Among A/H1N1 viruses circulating in Novosibirsk region three viruses contained four amino acid replacements in antigen sites Ca, Cb and Sb. In A/ H3N2 viruses from Novosibirsk, 2 amino acid substitutions were detected in antigen sites B and E. Based on genotyping influenzae epidemic on February to April of 2009 in the south of western Siberia was associated with influenza viruses A/H1N1, A/H3N2, and B. All A/H3N2 influenza virus isolates were variants of reference A/Brisbane/10/2007(H3N2) and A/ H1N1 influenza viruses isolates were similar to reference A/Brisbane/59/2007(H1N1).
    Zhurnal mikrobiologii, epidemiologii, i immunobiologii 01/2010;
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    ABSTRACT: The levels of susceptibility to influenza virus A/Aichi/2/68 H3N2 and the virus yield were determined using primary cells of the trachea and lungs of CD-1 mice and Wistar rats, and for 3 sets of cells obtained from primary lung cells of the both species by centrifugation in the gradient of density and by sedimentation on a surface. The values of ID50 virus dose for 10(6) cells and virus yield per 1 infected cell determined for primary mice cells were 4.0+/-0.47 and 3.2+/-0.27 IgEID50 (lung cells), 3.8+/-0.17 and 3.3+/-0.20 IgEID50 (tracheal cells), and those determined for primary rat cells were 4.0+/-0.35 and 2.1+/-0.24 IgEID50 (lung cells), 3.7+/-0.27 and 2.2+/-0.46 IgEID50 (tracheal cells). The values of ID50 and yield measured for mixtures of cells obtained from primary lung cells by centrifugation in gradient of density and by sedimentation on a surface differed insignificantly (p = 0.05) from the values of the corresponding parameters measured for lung and tracheal cells for both rats and mice. The analysis of data on the variation of the concentrations of different cell types in the experimental cell mixtures shows that type 1 and 2 alveolocytes possess significantly lower (p = 0.05) susceptibility and productivity vs. ciliated cells of the both species. The investigation was conducted within the frame of the ISTC/DARPA#450p project.
    Vestnik Rossiĭskoĭ akademii meditsinskikh nauk / Rossiĭskaia akademiia meditsinskikh nauk 02/2008;
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    Doklady Biochemistry and Biophysics 01/2008; 420:108-11. · 0.32 Impact Factor
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    ABSTRACT: Evaluations of immune system of 155 patients with rubella and 90 contacts with patients were examined. Detection of viral genetic material in blood, urine, and nasopharyngeal swabs has been performed using RT-PCR method. Clinical diagnosis has been confirmed by RT-PCR in 114 (73.5%) patients. Changes of laboratory tests for rubella without clinical signs of the infection were observed in 20% of contacts. Complex ELISA- and PCR-assisted examination of patients can help to determine the stage of disease and characteristics of immune response. For differential diagnostic of rubella and other infectious diseases with exanthema it is rational to perform complex examination of patients using immunologic and molecular biologic methods.
    Zhurnal mikrobiologii, epidemiologii, i immunobiologii 01/2007;
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    ABSTRACT: Two outbreaks of rubella infections notified in the Tomsk and Kemerovo Regions were investigated. Two rubella virus strains from one patient in each outbreak were isolated and genetically characterized. Reverse transcription polymerase chain reaction was used to reveal partial E1 gene sequence at a length of 915 nucleotides. Analysis indicated that the rubella virus strains circulating in the West-Siberian region belonged to international genetic 1g group, which had been first detected in Russia.
    Voprosy virusologii 01/2007; 52(2):16-9.
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    ABSTRACT: Twenty one strains of rubella virus were isolated in the Western Siberia during 2004-2006 epidemic period. Genotyping of isolated strains was performed by partial sequencing of glycoprotein E1 gene. Phylogenetic analysis showed that 20 out of 21 isolated in the Western Siberia strains of rubella virus belonged to genotype 1g, and 1 strain (isolated in Altai region in 2006)--to genotype 1E.
    Zhurnal mikrobiologii, epidemiologii, i immunobiologii 01/2007;
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    ABSTRACT: The mechanisms of infection development in intraperitoneal inoculation of mice by ectromelia virus strain K-1 and cowpox strain EP-2 were studied. Ultrastructural parameters of virus assembly and maturation are described. Differences in the types of cells replicating the viruses and in the type of visceral injuries were detected. The studies showed a local type of strain EP-2 cowpox infection and dissemination of ectromelia strain K-1.
    Bulletin of Experimental Biology and Medicine 05/2006; 141(4):448-52. · 0.34 Impact Factor
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    ABSTRACT: The study demonstrates the effects of kenalog (Kn), a synthetic glucocorticoid hormone, on the course of virus A/Aichi/2/68 influenza in white mice. In doses of 5 and 10 mg/kg, Kn reduced the weight of the adrenal glands, thymus and spleen, which was accompanied by decrease of the resistance to the mentioned virus, judging by LD50 decrease vs. this index in the control infected group. Besides, four days after infecting with 5 LD50 of influenza virus (IV), lung virus and interferon titers were significantly lower in mice pretreated with Kn vs. mice treated with placebo. Lung cell susceptibility to IV in vitro was identical in mice treated with Kn or placebo. In ultrathin lung sections of IV-infected mice, both experimental and control ones, there was virus budding in bronchial epithelium cells and type I and II alveolocytes. Analysis of inflammatory effusion compound in semithin lung sections 6 days after IV infection, found a substantially smaller number of mature alveolar macrophages (AM) and a bigger number of neutrophiles vs. infected controls. The authors reckon that higher mortality of mice pretreated with Kn before infecting, is caused not by enhancement of IV reproduction in target lung cells during influenza development, but by the contribution of other pathogenic factors. One of those may be increase of neutrophilic migration into the lungs; neutrophiles are more able to realize their significant destructive potential under the condition of reduction in the clearing function of AM and IV infection.
    Vestnik Rossiĭskoĭ akademii meditsinskikh nauk / Rossiĭskaia akademiia meditsinskikh nauk 02/2006;
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    ABSTRACT: The specific features of reproduction of EP-2 strain of cowpox virus (CPV) were studied in intranasally infected BALC/C mice by light and electron microscopy. Virus replication was found in the ciliated, intercalary, basal, and goblet cells (the nasal respiratory area), basal and supporting cells (the nasal olfactory area), ciliated, intercalary, goblet cells (the tracheal and bronchial epithelium), and collagen-producing, Schwann's, endothelial, smooth muscle, and adventitial cells. It has been shown that the CPV strain EP-2 locally replicates in the nasal cavity, trachea, and large bronchi and that there is no generalized infection.
    Voprosy virusologii 01/2005; 50(4):37-42.
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    ABSTRACT: The purpose of the case study was to evaluate comparatively the relative contribution of cell susceptibility and the inhibiting effect of factors of pulmonary epithelial lining in mice and rats to influenza virus A/Aichi/2/68 (H3N2) adapted to mice as related with the development of infection process in the lungs of experimental animals when infected in vivo and in vitro. Mice and rats were infected aerogenically with different doses of influenza virus. The primary cell-culture suspensions sampled from the lungs of mice and rats were used to study the adsorption and dynamics of influenza virus production in infection by different dose of influenza virus in vitro. The cell suspensions were shown to be able to produce the influenza virus for as long as 48 hours after infection. It was for the first time that the results denoted the identical susceptibility of primary pulmonary cells in mice and rats to influenza virus. A lower pulmonary susceptibility to influenza virus in rats versus mice could be indicative of that the surface factors of epithelial lining contribute essentially to shaping the pulmonary susceptibility to influenza virus since there is no difference of the susceptibility of pulmonary cells to influenza virus between the two above animals' species.
    Vestnik Rossiĭskoĭ akademii meditsinskikh nauk / Rossiĭskaia akademiia meditsinskikh nauk 02/2004;
  • ЗДРАВЭКСРО—СИБИР 2001. “ИННОВАЦИИ В ОХРАНЕ ЗДОРОВЬЯ ЛЮДЕЙ”. СБОРНИК ТЕЗИСОВ НАУЧНО-ПРАКТИЧЕСКОЙ КОНФЕРЕНЦИИ [ZDRAVÈKSRO—SIBIR 2001. “Innovations in public health”. Collection of abstracts of the scientific-practical conference], Novosibirsk, Novosibirsk Oblast, Russia; 11/2001
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    ABSTRACT: The specificity of lethal infection was studied in noninbred white mice (age--15 to 20 and 25 to 30 days) infected intraperitoneally with the EP-2 strain of cowpox virus (CPV) in doses 10(5), 10(6) and 10(7) PFU. The virus caused the lethal infection in the 15-20-day mice; while the 25-30-day mice remained healthy and survived. Virologic, immunologic-and-histochemical and electron-microscopy examinations of the 15-20-day mice revealed a replication of the EP-2 strain in tissues bordering on the virus introduction area; there was no generalization of infection. The virus replicated first in the mesothelium cells, and after that, in fibroblasts as well as in the endothelial, fatty, adventicial, cross-striated and muscle cells and in myosatellites.
    Voprosy virusologii 48(5):34-8.
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    ABSTRACT: Injections of exogenous DNA combined with a cytostatic agent cyclophosphamide (CP) cause illness and death in experimental mice. This phenomenon is referred to as delayed death. It has been found that fragments of exogenous DNA reach the bone marrow and enter the bone marrow cells (BMCs) 1–5 min after injections. Fragments of exogenous DNA are captured from culture medium by BMCs generated ex vivo. After joint incubation with BMCs of mice, the fragments of exogenous DNA are internalized into internal compartments in a nondegraded form. Up to 1800 kb of nucleic acid material can be present in each cell of BMCs generated ex vivo and treated with fragments of exogenous DNA. The fragments of exogenous DNA internalized in BMCs generated ex vivo of both intact and CP pretreated mice become circularized. In the case of intact mice, the fragments of exogenous DNA can form high-molecular weight structures in vivo. It is suggested that the exogenous fragments localized in BMC nuclei integrate into chromosome(s) of the recipient mouse genome when treated with CP and exogenous DNA.
    Russian Journal of Genetics: Applied Research. 2(6).
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    ABSTRACT: A molecular epidemiological study of novel strain of Rubella virus isolated during the outbreak in Western Siberia in 2004 is described. A detailed phylogenetic analysis performed based upon entire SP-section, which encodes all three Rubella structural proteins (C, E2, and El), is implemented. This analysis provides characterization of this strain and classifies it in the 1H genotype, thereby correcting a previous classification of this strain based upon a shorter nucleotide sequence only encoding El protein. Therefore, this study identified the genotype of the Rubella virus not previously detected in western Siberia (and even the entire Russian Federation), which highlights the importance of more extensive characterization of genetic variability of the Rubella virus, especially with regard to the potential influence of vaccination on the Rubella virus mutagenesis.
    Molecular Genetics Microbiology and Virology 26(4). · 0.27 Impact Factor