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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Vyakhireva JV, Filatova AYu, Krivosheeva IA, Skoblov MYu

RNA-interference enjoys a wide range of applications in medical and biological research. In particular, it is used to study functions of genes. One of the most popular approaches to this task is gene knockdown by small interfering RNA (siRNA). Currently there is no unified protocol for this method, which results in low reproducibility of experimental data. In the following article we outline the theoretical bases for this method and provide practical recommendations for its use in siRNA-mediated gene silencing experiments.

Received: 2017-06-25

Published online: 2017-07-19

DOI: 10.24075/brsmu.2017-03-02

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VIEWS 5552

Filatova AYu, Sparber PA, Krivosheeva IA, Skoblov MYu

As of today, there have been about 2,500 gene therapy clinical trials initiated or completed worldwide. Some of the tested drugs have been already approved for clinical use. Most of these drugs target well-characterized protein-coding genes. At the same time, the past few years have witnessed an increasing interest in long noncoding RNAs (lncRNAs) and their role in cellular processes. Of 16,000 identified human lncRNA genes, biological functions have been elucidated for only two hundred. Nevertheless, we already know about their association with the development of 200 different disorders. In some cases these genes are the key element in disease pathogenesis, which makes long noncoding RNAs a promising target for gene therapy. To date, researchers successfully employ molecular biology techniques for the development of lncRNA-based therapeutic strategies. The following review focuses on the main approaches to gene therapy based on the use of lncRNA.

Received: 2017-06-25

Published online: 2017-07-18

DOI: 10.24075/brsmu.2017-03-01

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VIEWS 5180

Guglya EB, Kotlobay AA, Sekretova EK, Volkova PV, Yampolsky IV

An extensive collection of plants gathered in the European part of Russia was screened for a substrate of fungal luciferase. This work was inspired by the recently discovered mechanism of bioluminescence in higher fungi and the structural similarity of fungal luciferin with some plant metabolites. Of all studied leaf extracts obtained from 200 different plants, bioluminescent activity was discovered in 10 species. Each of these species contained a plurality of active compounds. All the luminescent substrates were not identical to fungal luciferin (3-hydroxyhispidin) and were chemically unstable, rendering the attempt to isolate individual compounds for further structural characterization yet unsuccessful. This study is the first step towards engineering a self-luminescent plant based on a fungal enzyme-substrate bioluminescent system.

Received: 2017-02-19

Published online: 2017-06-04

DOI: 10.24075/brsmu.2017-02-10

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VIEWS 4978

Malov IA, Strenev NV, Takhchidi KhP

Premacular hemorrhage occurs in various disorders and causes sudden unilateral or bilateral visual impairment. One of the well-established techniques to treat this condition is Ng:YAG laser hyaloidotomy. Below we report a case of premacular hemorrhage in the right and left eyes of a 23-year old patient with acute myeloblastic leukemia. Ng:YAG laser hyaloidotomy was successfully performed on both patient’s eyes at different puncture sites.

Received: 2017-04-10

Published online: 2017-06-02

DOI: 10.24075/brsmu.2017-02-09

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VIEWS 4600

Tkacheva SV, Manapova ER, Sozinova YuM, Yakupova FM, Fazylova YuV

Interferon-based regimens for chronic hepatitis C (HCV) are quite common, just like interferon-free treatments, and are extensively used in Russia because interferon is widely available to most patients. In 2013 the original Russian drug cepeginterferon alpha-2b (cepegIFN alpha-2b marketed as Algeron by Biocad, Russia) was introduced into clinical practice. The aim of this study was to assess effectiveness and safety of cepegIFN alpha-2b as part of the combination therapy with ribavirin in patients with chronic HCV infection. The study was conducted over the period from 2014 to 2016 and recruited 37 patients with chronic genotype 1 HCV infection: 22 men and 15 women (mean age of 42.0 ± 5.2 years). All of them received the following combination antiviral therapy (AT): 1.5 μg/kg cepegIFN alpha-2b once a week and 15 μg/kg ribavirin daily over the period of 48 weeks. Effectiveness of AT was assessed by the rate of sustained virological response (SVR), i. e. aviremia achieved 24 weeks after the onset of treatment. In our SVR was observed in 26 patients (70.3 %). Adverse effects seen in the course of AT were typical of interferon-based drugs and ribavirin. CepegIFN alpha-2b dosage was corrected in two patients who developed neutropenia; ribavirin dosage was corrected in 3 patients who developed anemia. Based on the obtained results, we recommend including cepegIFN alpha-2b into the combination antiviral therapy in patients with chronic HCV infection.

Received: 2017-02-09

Published online: 2017-06-02

DOI: 10.24075/brsmu.2017-02-08

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VIEWS 4720

Voroshilina ES, Plotko EE, Khayutin LV, Tischenko NA, Zornikov DL

Moderate vaginal dysbiosis is a shift in normal vaginal microbiota composition characterized by increased levels of opportunistic microbes and an ordinary high proportion of lactobacilli that make up 20 to 80 % of the total microbial population of the vagina. Some women with vaginal dysbiosis do not show any symptoms of the infectious inflammatory condition (IIC), which raises the question of whether their dysbiosis should be corrected. We studied the association between some parameters of the microbiota and clinical symptoms of IIC in female patients with moderate vaginal dysbiosis. Participants were distributed into two groups: group 1 included patients with clinical symptoms of IIC (n = 91), group 2 was comprised of asymptomatic patients (n = 44). Mean age was 26.9 ± 6.9 years. Vaginal microbial communities were studied using real-time polymerase chain reaction assays. Levels of six Lactobacillus species were measured in the vaginal discharge: Lactobacillus crispatus, L. iners, L. jensenii, L. gasseri, L. johnsonii, and L. vaginalis. We found that L. iners dominated the microbiota of 45 (49.5 %) symptomatic patients and only 9 (20.5 %) asymptomatic individuals (p = 0.002), unlike L. gasseri that significantly prevailed in the samples of asymptomatic patients: 23 (52.3 %) women vs 21 (23.1 %) in the group of patients with clinical signs of IIC (p = 0.001).

Received: 2017-04-09

Published online: 2017-06-01

DOI: 10.24075/brsmu.2017-02-07

Comments 0

VIEWS 5070

Voroshilina ES, Zornikov DL, Plotko EE

Criteria of normality for the vaginal microbiota of healthy women are still a subject of discussion. A decision to assign a study participant to a group of healthy individuals is quite subjective if based on the absence of complaints and physical examination only, which renders study results ambiguous. Below we compare occurrence of the normal vaginal flora and vaginal dysbiosis in women divided into 3 groups according to the examination type (patient’s subjective evaluation of her condition, physical examination, and laboratory tests). We examined 234 women of reproductive age from Yekaterinburg (mean age was 30.3 ± 6.6 years). Microbiota composition and lactobacillus diversity (L. crispatus, L. iners, L. jensenii, L. gasseri, L. johnsonii, L. vaginalis) were evaluated by real-time polymerase chain reaction using the Femoflor assay and reagent kits by DNA-Technology, Russia. One in 5 women of reproductive age who had no health complaints was found to have dysbiosis. The normal microbiota of those women was dominated mostly by L.iners, while dominant L. crispatus were observed in every third participant. Prevailing L. crispatus were also found in the normal microbiota of 46.2 % of women who were considered healthy based on the doctor’s examination and laboratory tests. Thus, clinical evaluation of the female lower reproductive tract can be compromised by doctor’s subjectivity if not supported by laboratory tests and may overlook vaginal dysbiosis in the patient.

Received: 2017-04-11

Published online: 2017-05-31

DOI: 10.24075/brsmu.2017-02-06

Comments 0

VIEWS 5060

Pochernikov DG, Galkina IS, Postovoytenko NT, Gerasimov AM

Many sexually transmitted diseases are caused by bacteria. While we fairly well understand the role of some microorganisms in the development of genitourinary tract infections, there is still a vast majority of those whose contribution is unclear. It is believed that sexual partners share their genitourinary microbiota, meaning that treatment regimens should be the same for both of them. This article reports results of the study of seminal and cervical microbial communities conducted in 50 married couples who did not use barrier birth control and did not take any antibiotics at least 3 months before the study. All couples presented with complaints of primary or secondary infertility, recurrent miscarriages or sought preconceptional counseling. The mean age of male and female participants was 34.8 ± 7.8 and 30.4 ± 6.2 years, respectively. Samples of the seminal fluid and vaginal secretions were studied by real-time polymerase chain reaction (real-time PCR) with Androflor and Femoflor reagent kits. The following bacteria were more frequent in the vaginal microbiota than in the seminal fluid: Lactobacillus spp. (p < 0.005), Eubacterium spp. (p = 0.002), Gardnerella vaginalis (p = 0.002), Megasphaera spp./Velionella spp./Dialister spp. (p = 0.004). Ureaplasma spp. was 3 times more frequent in women, Mycoplasma hominis was 4 times more frequent in men; however, this difference was not significant. In 4 (8 %) couples both partners had normal microbiota; 23 (46 %) couples shared at least one microbiota resident. Also, microbial communities were totally different in 23 couples. The obtained data indicate that both sexual partners should be examined to decide on the most effective treatment for each of them. Qualitative and quantitative real-time PCR assays Androflor and Femoflor provide comprehensive data essential for adequate treatment planning.

Received: 2017-04-08

Published online: 2017-05-31

DOI: 10.24075/brsmu.2017-02-05

Comments 0

VIEWS 5204