Bulletin of RSMU

ISSN Print 2500–1094 ISSN Online 2542–1204

Bulletin of RSMU

BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)

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Belous AS, Shevelev AB, Trubnikova EV, Biryukova YuK, Mishina ES, Loyko EA, Lebedeva AA, Zakharchenko NS

Management of purulent wounds is a problem that requires particular attention: wounds are a common injury type for which suppurative complications are frequent, mortality rates are high and antimicrobial therapy may be ineffective due to the presence of drug-resistant bacteria in the wound. In this work we have studied the effectiveness of wound treatment with the leaf extract of transgenic Kalanchoe pinnata modified to produce antimicrobial peptide cecropin P1. Purulent wounds infected with Staphylococcus aureus were modeled in Wistar rats. Four groups of animals were formed, with 10 animals in each group. In all groups, the wounds were cleansed with 3 % hydrogen peroxide solution once a day; all groups except the controls received additional treatment. Group 2 received 10 % cefazolin solution, group 3 received kalanchoe juice, group 4 received the juice of cecropin P1-producing kalanchoe. Histologic stains of biopsy samples were performed after rats were sacrificed by anesthetic overdose on days 3, 10 and 14 after treatment onset. On day 3, wound dynamics was the same in all groups. On day 10 exudate was still observed in the controls; in group two exudation was almost finished and regeneration was about to begin; in groups 3 and 4 the wound defect was filled with granulation tissue. In spite of epidermal repair along the wound edges in groups 2 and 3, there still was some sloughing and granulation tissue was less mature than in group 4. We recommend conducting more extensive clinical research of the leaf extract of cecropin P1-containing transgenic Kalanchoe pinnata.

Received: 2017-02-05

Published online: 2017-03-14

DOI: 10.24075/brsmu.2017-01-09

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VIEWS 4509

Eremeeva ZhG, Fazylov VH

Individuals carrying occult (latent) hepatitis B pose epidemiological threat. Testing donated blood donors for surface antigen HBsAg (hepatitis B virus, HBV) only does not allow to assume the blood safe from the point of view of infections, which can result in post-transfusion transmission of infection. Lack of confidence here is due to the fact that the virus is present in the body even when HBsAg is negative. The study analyzes data of 61,155 blood donors of the Republican Blood Center (Kazan), collected in 2010–2014. The tests applied were those aimed at detecting HBsAg, anti-HBc-total, anti-HBc IgM (enzyme immunoassay), and determining DNA of the virus in the blood by polymerase chain reaction in "real time". It was found that donors with occult hepatitis B are identified each year, but their numbers decrease gradually. To prevent the spread of the virus it is recommended to add the anti-HBc-total test to the standard set of diagnostic tests.

Received: 2017-02-01

Published online: 2017-03-14

DOI: 10.24075/brsmu.2017-01-08

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VIEWS 4522

Oleynik AF, Fazylov VH, Beshimov AT

Antiretroviral therapy (ART) for HIV-positive patients allowed labeling the disease a therapeutically controlled one. The main goal of ART is to prolong patient's life and preserve its quality. This is accomplished through viral load reduction (decrease of the number of HIV-RNA copies in blood plasma), which leads to the growing numbers of CD4<sup>+</sup>-T-lymphocytes. However, ART can be ineffective. In 2010–1014, we conducted an observational cohort retro/prospective study aimed at learning how often ART can be ineffective from immunological (II), virological (VI) and clinical points of view. The study was carried out at the premises of the Republic Center of AIDS and Infectious Diseases (Kazan, Russia). The study included 341 adult HIV-positive patients subjected to ART at 3rd and 4th stages of disease's development, with the treatment virologically efficient at least during the first year. The observation period was 1 to 3 years. ART was considered II (immunologically inefficient) when the number of CD4<sup>+</sup> increased for less than 50 cells/mcl through the year with HIV completely suppressed. VI (virological inefficiency) of ART was registered if the number of HIV RNA copies was above the definition threshold after 6 months of treatment. ART was II in 14.0–15.9 % of cases after a year of treatment and in 22 % of cases after three years. It was noted that subsequent restoration of an adequate number of T-lymphocytes CD4<sup>+</sup> required they overcame the threshold of 100 cells/mcl within the 1st year of treatment. Virologically, ART was effective for 92.7 % for patients. Most (80 %) cases of VI of ART were results of patients' lax attitude towards treatment. Clinically, ART helped 91 % of patients; this result largely depended on the number participants for whom ART was II. II of ART is a risk factor, the risk being progression of the disease with active ART in the background and death of the HIV-positive individual. II of ART makes the risk of clinical progression of HIV 6.232 times higher (95 % CI 3.106–12.51).

Received: 2017-01-31

Published online: 2017-03-13

DOI: 10.24075/brsmu.2017-01-07

Comments 0

VIEWS 5019

Chernyavskaya OA

Increasing HIV prevalence among women and growing numbers of HIV-positive patients who choose to become pregnant prompt a discussion of management strategies applied to such patients. In this work we analyze a case of a pregnant HIV-positive woman with marked immunodeficiency who started seeking medical advice only after she had developed severe life-threatening secondary conditions. We look at the progression of comorbidities that led to the death of the patient and her baby and evaluate the chosen treatment plan. We also propose recommendations for the management of patients with similar pathologies that include psychological care, vigilance against possible atypical progression of a comorbidity, such as tuberculosis, and extensive diagnostic evaluation.

Received: 2017-02-04

Published online: 2017-03-13

DOI: 10.24075/brsmu.2017-01-06

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VIEWS 4922

Shur KV, Maslov DA, Bekker OB, Danilenko VN

Antibiotic selection pressure, genetic polymorphism as well as diversity of the immune status of the host and other selection factors continuously prompt Mycobacterium tuberculosis, the tuberculosis causative agent, to evolve. Significant or insignificant mutations shape new (sub)lineages of the pathogen whose evolution can be understood only through analyzing and monitoring its genotypic diversity and properties of its lineages. In our study we used a set of 46 M. tuberculosis clinical isolates from Moscow region. The samples were typed using the standard 24-loci MIRU-VNTR technique. Beijing family isolates were shown to prevail in the collection (60.9 %), as well as Beijing-B0/W148 subtype (60.7 % of total Beijing type samples); most of them (88,2 %) were multidrug-resistant resistant. The applied technique allowed us to detect one case of a mixed-strain infection.

Received: 2017-02-01

Published online: 2017-03-13

DOI: 10.24075/brsmu.2017-01-05

Comments 0

VIEWS 4828

Maslov DA, Bekker OB, Alekseeva MG, Kniazeva LM, Mavletova DA, Afanasyev II, Vasilevich NI, Danilenko VN

Tuberculosis (TB) is the world’s deadliest bacterial infection. Its causative agent Mycobacterium tuberculosis evolves into rapidly spreading multidrug-resistant and extensively drug-resistant (MDR and XDR) strains, which complicates the treatment. Therefore, the use of novel target-specific chemical compounds is crucial for the development of effective antituberculosis agents. Serine/threonine protein kinases (STPKs) of M. tuberculosis are currently considered as attractive drug targets. In turn, aminopyridines and aminopyrimidines that have not been used for TB treatment so far exhibit inhibitory activity towards STPKs. In this study we screened 192 aminopyridine- and aminopyrimidine-based compounds using the Mycobacterium smegmatis aphVIII+ test system designed to screen for active STPKs inhibitors. First, we selected 53 compounds with subinhibiting concentrations of up to 100 nmol/disk. Of them, 22 showed STPKs-inhibiting activity in the test system, which was confirmed in vitro on the M. tuberculosis PknA protein with a maximum of 26.9 ± 6.1 %. Toxicity testing was performed in vitro on human embryo fibroblasts using the MTT-assay. Ultimately, 3 relatively active and relatively non-toxic STPKs inhibitors were selected for further research as drug candidates for MDR-TB treatment.

Received: 2017-01-30

Published online: 2017-03-13

DOI: 10.24075/brsmu.2017-01-04

Comments 0

VIEWS 5099

Chagina IA, Perevarova YuS, Perevarov VV, Chaplin AV, Borisova OYu, Kafarskaya LI, Afanas'ev SS, Aleshkin VA

The pathogenic mechanism used by Corynebacterium diphtheriae is attributed to the ability of the diphtheria toxin to disrupt protein synthesis in human cells. Diphtheria toxin production is regulated by the DtxR protein. The latter is involved in the iron-mediated repression of the toxin gene and coordinates activities of other genes essential for the survival of C. diphtheriae. The DtxR-encoding gene occurs in both toxigenic and non-toxigenic strains; therefore it can be used to analyze the population structure of the species. In our work we have studied 45 strains of C. diphtheriae isolated in the Russian Federation in 2010–2015. These strains were analyzed to reveal that gene dtxR is a highly conservative region of С. diphtheriae genome that can be found in all members of the studied species. The majority of the discovered polymorphisms were synonymous (16 of 18 single nucleotide polymorphisms identified). In spite of the low phylogenetic signal, the allelic variant of dtxR was associated with the strain’s phenotype (biovar, toxigenicity). The obtained data indicate the presence of aggressive negative selection aimed to maintain the existing protein sequence in the population. Based on the results, we recommend dtxR polymerase chain reaction as an additional technique for pathogen identification, which is especially relevant considering the increasing prevalence of the disease associated with non-toxigenic C. diphtheriae strains.

Received: 2017-02-02

Published online: 2017-03-11

DOI: 10.24075/brsmu.2017-01-03

Comments 0

VIEWS 4628

Gordeev AB, Lyubasovskaya LA, Rodchenko JV, Dubodelov DV, Mukosey IS, Kochetkova TO, Nikitina IV, Ionov OV, Zubkov VV, Trofimov DYu, Priputnevich TV

Staphylococcus epidermidis is a member of the normal bacterial flora of humans capable of causing potentially dangerous diseases in neonates with very or extremely low birth weight. The number of genes responsible for virulence and antibiotic resistance may vary in different S. <i>epidermidis</i> strains. We sequenced isolates of S. <i>epidermidis</i> to explore genetic diversity of 14 strains circulating in the Neonatal Intensive Care Unit of Kulakov Research Center for Obstetrics, Gynecology and Perinatology. Among the studied strains, 8 sequence types were identified, the most frequent being ST2 and ST59, both of which belong to the clonal complex CC2. Of 14 studied strains, 10 were of CC2 type. The studied strains revealed a variety of genes responsible for antibiotic resistance. We found 15 genes that provided resistance to aminoglycosides, beta-lactam antibiotics, fusidic acid, macrolides, lincosamides, streptogramin B, tetracycline, and trimethoprim. We identified a number of genes associated with virulence (<i>aae, atlE, aap, embp</i>), whose frequency in the studied isolates was varied. The insertion element IS256 was detected in 9 strains, and 7 strains revealed the presence of the <i>ica</i>-operon responsible for the biosynthesis of the biofilm matrix proteins.

Received: 2017-02-02

Published online: 2017-03-11

DOI: 10.24075/brsmu.2017-01-02

Comments 0

VIEWS 4619