[Frequency of drug resistance and immune escape mutations in the hepatitis B virus genome detected in pregnant women in the Republic of Guinea].

The aim of the work is to assess the prevalence of hepatitis B virus drug resistance mutations and immune escape mutations in pregnant women in the Republic of Guinea. MATERIALS AND METHODS: Blood plasma samples obtained from 480 pregnant women from different regions of the Republic of Guinea with laboratory-confirmed viral hepatitis B were studied. Nucleotide sequences for genotype identification and mutation detection were obtained using nested-PCR followed by Sanger sequencing, based on overlapping pairs of primers spanning the complete genome of the virus. RESULTS AND DISCUSSION: In the examined group, the viral genotype E was the most prevalent (92.92%) compared with subgenotypes A1 (1.67%), A3 (1.46%), D1 (0.63%), D2 (1.04%) and D3 (2.29%). Among the examined HBV-infected pregnant women, 188 (39.17%) had undetectable HBsAg. Drug resistance mutations were detected in 33 individuals, which amounted to 6.88%. The following mutations were found: S78T (27.27%), L80I (24.24%), S202I (15.15%), M204I/V (42.42%). The presence of polymorphic variants not described as drug resistant has also been shown in positions associated with the development of drug resistance to tenofovir, lamivudine, telbivudine and entecavir (L80F, S202I, M204R). When analyzing the MHR and the region of a determinant, mutations were detected in 318 (66.25%) of pregnant women. In 172 of them, which amounted to 54.09%, multiple mutations were found. The amino acid substitutions in 13 positions associated with HBsAg-negative hepatitis B and/or potentially affecting HBsAg antigenicity were identified. CONCLUSION: The high prevalence of immune escape and drug resistance mutations potentially associated with false-negative result of HBsAg screening, prophylaxis failure, and virological failure of therapy that has been identified among treatment naive pregnant women imposes a serious problem.

[Genetic diversity of the human immunodeficiency virus (HIV-1) in the Kaliningrad region].

INTRODUCTION: As is currently known, the epidemic process in the Kaliningrad Region was mainly associated with the spread of the recombinant form of HIV-1 (CRF03_AB); however, regular HIV importations from other countries and continents has created favorable conditions for emergence and spread of various recombinant forms of the virus.The most complete information on the diversity of recombinant forms in the region is also necessary to understand the structure of drug resistance (DR). The aim of the study was to explore the HIV-1 genetic diversity in the Kaliningrad Region. MATERIALS AND METHODS: We studied 162 blood plasma samples obtained from patients from the Kaliningrad Region, both with confirmed virological failure of antiretroviral therapy (ART) and with newly diagnosed HIV infection. For reverse transcription and amplification of HIV genome fragments, diagnostic «AmpliSense HIVResist-Seq¼. RESULTS AND DISCUSSION: The various recombinants between subtypes A and B (74%) were predominant in study group: recombinant was between CRF03_AB and subtype A (33.95%) and CRF03_AB-like (13.58%) were the most common. Among the "pure" subtypes of the virus, subtype A6 (16.67%). The circulation of subtypes B (3.70%) and G (1.23%) was also noted.Ninety-six patients (59.26%) were identified with at least one mutation associated with antiretroviral (ARV) drug resistance. CONCLUSION: The observed diversity of subtypes and recombinant forms of the virus implies that the new recombinants are actively emerging in the studied region, both between existing recombinant forms and "pure" subtypes, as well as between "pure" subtypes.

[Prevalence of viral hepatitis B markers among blood donors in the Republic of Guinea].

INTRODUCTION: The problem of transfusion safety in relation to parenteral viral hepatitis still remains relevant. Viral hepatitis B (HB) remains the most common viral infection transmitted through transfusion procedures. One of the natural phases of chronic hepatitis B (CHB) is occult hepatitis B infection (OBI), characterized by an undetectable HBsAg (regardless of the other serological markers content) in the presence of hepatitis B virus (HBV) DNA in the liver tissue and an extremely low, up to undetectable, level of viral load in the blood. In the Republic of Guinea, as in most countries on the continent, the prevention of HBV transmission through transfusion is still based on HBsAg serological testing of donors only. In this connection, OBI remains as a potential threat to blood transfusion safety. Detection of HBV DNA is a reliable preventive measure against transmission of the virus from donors with HBsAg-negative HBV infection, especially in highly endemic regions. In this regard, the study was conducted to substantiate recommendations for improving blood safety against the background of significant HBV prevalence in the Republic of Guinea.The aim of the work was the evaluation of serological and molecular markers of HBV infection in blood donors in the Republic of Guinea. MATERIAL AND METHODS: We examined 250 blood samples obtained from donors living in Conakry, Republic of Guinea. Samples were tested for the presence of serological (surface antigen, HBsAg; antibodies (ABs) to surface (anti-HBs IgG) and core (anti-HBc IgG) antigens) and molecular (DNA) markers of HBV infection. RESULTS AND DISCUSSION: The overall detection rate of hepatitis B markers was 83.2%; HBsAg was detected in 16.4% of all individuals. The high incidence of HBsAg in men (19.55%) compared to women (8.45%) was shown, the relative risk of HBV infection with the formation of HBsAg-positive chronic hepatitis B in males was also significantly higher. The prevalence of the HBV DNA in the study group was 30.4%, the OBI cases accounted for 15.6%. The prevalence of this form of the disease was shown in donors aged 30-49 years (24.78%), in the group of people younger than 30 years, the incidence was lower (8.73%), and at the age of over 50 years, OBI was not detected. Based on the phylogenetic analysis of 76 virus isolates, it was shown that genotype E prevails in the examined group (85.53%).Cases of pathogen DNA detection occurred in HBsAg-negative blood donors in the presence of anti-HBs IgG (n = 4), as well as in the simultaneous presence of ABs anti-HBs IgG and anti-HBc IgG (n = 7). The viral load exceeded 200 IU/ml in OBI samples. Escape mutations were detected by sequencing in each OBI sample, contributing to the virus escaping from diagnostic based on screening for HBsAg. CONCLUSION: Assessment of the prevalence viral hepatitis B markers in blood donors, determination of genotypes and clinically significant mutations of virus variants are necessary to ensure safe medical manipulations, control and prevention of the spread of this infectious agent.

Serological diagnosis and prevalence of HIV-1 infection in Russian metropolitan areas.

BACKGROUND: HIV infection is a major health problem in Russia. We aimed to assess HIV prevalence in different population groups and to compare the characteristics of 4th generation immunoassays from Abbott, Bio-Rad, Vector-Best, Diagnostic Systems, and Medical Biological Unit. METHODS: The study included 4452 individuals from the general population (GP), 391 subjects at high risk of HIV infection (HR) and 699 with potentially interfering conditions. HIV positivity was confirmed by immunoblot and by HIV RNA, seroconversion and virus diversity panels were also used. HIV avidity was employed to assess recent infections. RESULTS: The prevalence in GP was 0.40%, higher in males (0.62%) and in people aged < 40 years (0.58%). Patients attending dermo-venereal centers and drug users had a high prevalence (34.1 and 58.8%). Recent infections were diagnosed in 20% of GP and in 4.2% of HR. Assay sensitivity was 100% except for one false negative (99,54%, MBU). Specificity was 99.58-99.89% overall, but as low as 93.26% on HR (Vector-Best). Small differences on early seroconversion were recorded. Only the Abbott assay detected all samples on the viral diversity panel. CONCLUSION: HIV infection rate in the high-risk groups suggests that awareness and screening campaigns should be enhanced. Fourth generation assays are adequate but performance differences must be considered.

Optimization of the algorithm diagnosis chronic hepatitis B markers in patients with newly diagnosed HIV infection.

The possibility of modifying the algorithms for chronic viral hepatitis B laboratory diagnosis in individuals with newly diagnosed HIV infection is analyzed. Plasma samples were used from 196 patients residing in the Northwestern Federal District. Serological HBV markers were found in 79.6% of cases. However, HBsAg was detected in 5.6% of patients. Anti-HBcore IgG antibodies are found in 62.24% of cases, anti-HBe IgG antibodies in 27.55%, anti-HBs IgG antibodies in 52.55% of cases. Using a commercial kit with a 100 IU / ml sensitivity, HBV DNA was detected in 4.6% of patients, that is, 81.8% of HBsAg-positive individuals. Using the method developed by us, HBV DNA was found in 18.36% of HIV-infected individuals, including 12.75% of cases was HBsAg-negative (latent) disease form. In the examined group, HBV of genotype D prevailed (91.7%), genotype A was detected in 8.3% of cases. The distribution of subgenotypes is presented in the following ratios: D2 - 55.6%, D1 - 22.2%, D3 - 13.9%, A2 - 8.3%. Mutations were detected in the reverse transcriptase (RT) region in 91.6% of patients, in the SHB region in 83.3%, in the Core and Precore regions in 72.2% and in 27.7% of patients, respectively. Three HBV isolates (8.3%) were identified with drug resistance mutations to lamivudine, entericavir, telbivudine and tenofovir, which are amino acid substitutions in the HBV polymerase gene at positions L180M, T184A, M204V. Vaccine escape mutations were detected in 61.1% of patients. In all samples with drug resistance mutations, escape-mutants were simultaneously present. When analyzing the basal nucleus promoter, Precore and Core regions, 22.2% of patients with the double mutation A1762T / G1764A, 25% with the mutation G1896A were identified. In one person, all three substitutions were found. In the Core region, 77.7% of patients showed mutations in one of the hot spots (codons 87, 97, 112, and 130 substitution), which can play a role in immunomodulation in CHB. Analysis of the HBV genetic structure, mutations detection early in the virus in patients with HBV can help predict the clinical course and disease progression, and ART complications. To reduce the HIV HBV co-infection burden and to appointer anti-HBV therapy, it is necessary to introduce detection the occult HBV to modify the algorithm for CHB laboratory diagnosis.

[The prevalence clinically significant virus mutations among patients with chronic viral hepatitis B.]

The prevalence of clinically significant virus mutations in patients with chronic viral hepatitis B from the Kyrgyz Republic was analyzed. Blood plasma samples of 64 patients with verified chronic viral hepatitis B obtained from Kyrgyzstan indigenous people were used in the work. Asymmetric PCR was carried out with extended oligonucleotides and the first reaction amplification product was further used in a new PCR with one of the nested pairs overlapping primers that flanked the entire HBV genome together, followed by sequencing. Based on the phylogenetic analysis of 64 HBV isolates obtained from patients from the Kyrgyz Republic, it was shown that only the genotype D virus was present in the examined group, the HBV subgenotype D1 (68.75%) prevailed compared with the HBV subgenotype D2 (18.75%) and subgenotype D3 (12.5%). For all subgenotypes, several independent infection sources are obvious, subclusters that include isolates from Kyrgyzstan, Kazakhstan and Uzbekistan are distinguished, as well as subclusters that include isolates only from Kyrgyzstan, which are less similar to isolates previously deposited in the international database, which probably indicates an independent HBV homologous evolution in the region. Clinically significant mutations were identified in 26.5% of patients. Including 12.5% with escape mutations that prevent the virus detection and / or allow the virus to replicate despite the vaccine (122K, 128V, 133I, 134N). Another 12.5% of the isolates are characterized by mutations that are independently associated with the liver cirrhosis and hepatocellular carcinoma development, including 21, 24, 27 nucleotides deletions in the Pre-S2 region and the S11F mutation in the PreCore region. In one case, unusual 236S and 250P mutations were found in the positions described as drug resistance sites of the P region associated with the resistance development to adefovir, tenofovir, and entecavir. The hepatitis B virus genetic structure analysis, early virus mutations detection in patients with chronic hepatitis B virus can help to choose the right vaccination strategy, antiviral and immunosuppressive therapy, as well as predict the clinical course and disease progression.

[Identification and molecular-genetic characteristics of the hepatitis B virus among HIV-infected patients in Arkhangelsk.]

AIM: To estimate the prevalence and characterize the hepatitis B virus among HIV-infected patients with virological failure of antiretroviral therapy in Arkhangelsk. MATERIAL AND METHODS: HBV markers determinations (HBsAg, anti-HBs IgG, antiHBcor IgG, DNA HBV) were performed in isolates from blood plasma samples 64 HIV-infected patients with virological failure of antiretroviral therapy (viral load >50 IU / ml after 6 months of antiretroviral therapy or an increase in viral load after primary suppression of viral replication). For the detection of the hepatitis B virus, nucleic acids were isolated using the commercial kit «AmplePrime Ribo-prep¼. The virus presence analysis was performing by the polymerase chain reaction (PCR) method with hybridization-fluorescence detection in "real time" using the commercial set of «AmpliSens® HBV-FL¼. In the future, we used the method developed by the Saint-Petersburg Pasteur Institute, which allows detecting HBV in biological material with a low viral load. RESULTS: HBsAg-negative (occult) HBV was detect in 28 (43.8%) HIVinfected patients. Only HBV genotype D was detected, and the HBV subgenotype D1 prevailed (39.3%) compared with the HBV subgenotype D2 (32.1%) and D3 (28.6%). Serological markers in 42.8% of patients with HBV DNA were founding. Two HBV isolates with drug resistance mutations in the polymerase gene, leaded to amino acid substitutions (L180M, M204V) associated with the resistance development to lamivudine, entecavir, telbivudine and tenofovir were identifying. CONCLUSION: The occult (HBsAg-negative) HBV high prevalence among HIV-infected patients suggests the need to use molecular-biological diagnostic methods to identify HBV, as well as to analyze the HBV drug resistance mutation before starting antiretroviral therapy for HIV.

[The identification of clinical strains Pseudomonas fulva using techniques of MALDI-TOF mass spectrometry and common analysis].

The technique MALDI-TOF mass spectrometry was applied using device Microflex with database MALDI Biotyper (Bruckeer Daltonics Inc.) to identify with high level of reliability 8 strains P. fulva from collection of pseudo monads isolated from clinical material in St. Petersburg. When analyzing the same strains applying technique MALDI TOF mass spectrometry using device Vitek MS (bioMerieux) these starins were wrongly identifies as P. putida. The complex of tests of common analysis was approved and proposed for control differentiation of P. fulva and P. putida. The medical significance of P. fulva was approved.

[Electrical pulse therapy for the treatment of the patients with obliterative atherosclerosis of the vessels in the lower extremities].

The authors report the results of the application of electrical pulses to the region of lumbar ganglia for the treatment of patients with obliterative atherosclerosis of the blood vessels in the lower extremities. It was shown that the temporary blockade of sympathetic regulation of the lower limb arteries promotes reduction of vascular tone in the distal segments of the lower extremities, stimulates collateral blood flow, and improves microcirculation in ischemic tissues(specifically, following reconstructive surgery on the lower limb arteries).

[Current state-of-the-art of rehabilitative treatment of chronic obliterative arterial diseases of the lower extremities].

Up-to-date approaches and methods for balneotherapy of chronic obliterative arterial diseases of the lower extremities are discussed with special reference to their confirmed clinical efficiency. The necessity of differential application of physical agents to the treatment and rehabilitation of the patients with these diseases is substantiated.

[Albumin differentiation by isoelectric focusing in patients with liver cirrhosis]. / Differentsirovka al'bumina metodom isoélektricheskogo fokusirovaniia u bol'nykh tsirrozom pecheni.

In patients with cirrhosis of the liver, there is noted a striking imbalance in the distribution of isoelectric fractions of serum albumin in the isoelectric field. Disproportion in isoelectric fractions of serum albumin can, we believe, be referred to functional loads on albumin relative to fixation and transport of surplus of bilirubin and ammonia. In the ascitic fluid, there comes to be seen, in fact, a reversal of imbalance of pH-dependent fractions of albumin,--probably, due to "purification" in the lymphatic system and partly in the functioning hepatocytes. As to the isoelectric spectrum, the ascitic fluid albumin is qualitatively very similar to serum albumin of healthy people.

[The inhibiting effect of cimetidine on a number of enzymes]. / Ingibiruiushchee vliianie tsimetidina na riad fermentov.

The results of the comparative study on the contents and activity of the enzymes of the system of oxidases of mixed function in the lungs and liver in rabbits, the specific features of phospholipid spectrum in the supernatant and microsomal fractions of the lungs and liver tissues are presented. The same parameters were investigated in experimental chronic inflammatory diseases of the lungs. The degree of the damage of the monoxygenase capacity of the lungs and the degree of the liver involvement in the process were established. The peculiarities of the inhibitory action of cimetidine on the detoxication capacity of the lungs and liver and on the disorder of the reproduction of phospholipids in these organs were revealed. The substrate specificity of cimetidine in inhibition of microsomal NADPN-cytochrome-C-reductase both in the liver and in the lungs, the probability of blocking the synthesis of phospholipids at the level of phosphatide acid and citidinephosphatecholine were noted. The results obtained should be taken into consideration during drug therapy.