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A number of drugs based on recombinant erythropoietin contain human serum albumin as an auxiliary component. The presence of this protein hinders the proper control of the drug quality in accordance with the requirements of regulating agencies. We propose the novel method for separation of recombinant erythropoietin (epoetin beta) and human serum albumin. It is based on the subsequent use of hydrophobic sorbent and anion exchange resin placed in gravity flow columns (without the use of spin-columns). The proposed approach makes it possible to concentrate and purify the preparations containing the epoetin beta both at high and at minimal concentrations (the ratio of the amount of albumin and erythropoietin in the used preparations can reach 125:1). The average yield of epoetin beta after the use of hydrophobic sorbent and anion exchange resin was 75 % and 97 %, respectively. It was shown that the determined conditions of sample preparation had no affect on the content of the epoetin beta in the product., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Zybin reports a relationship with Pharmapark LLC that includes: employment. Orlova reports a relationship with Pharmapark LLC that includes: employment. Klishin reports a relationship with Pharmapark LLC that includes: employment. Zyryanov reports a relationship with Pharmapark LLC that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Increasing HIV drug resistance is an important public health concern. The current study aimed to assess HIV drug resistance among people who live with HIV (PLWH) experiencing virological failure. Blood samples and epidemiological characteristics were collected in four Siberian regions from PLWH experiencing ART failure. Partial pol gene sequences were obtained for the study individuals. Drug resistance mutations (DRMs) were predicted using the Stanford HIVdb Program. The association of HIV DRM with epidemiological characteristics was estimated using logistic regression analysis. Further analysis was performed for children (0-14 y old) and adults (≥15 y old) separately. In total, 815 (89.4%) patients were included in the final dataset. Overall, 501 (61.5%) patients had DRM detected. NRTI DRM was more common in children, while NRTI+NNRTI DRM was more frequent in adults (P < 0.001). Krasnoyarsk region, male sex and high viral load were positively associated with the presence of DRM in adults, while higher CD4 cell count and PI/INSTI-based ART had a negative association. No association between epidemiological characteristics and DRM was identified in children. The remaining 38.5% of patients with virological failure had no DRM detected; those patients were likely to have insufficient ART adherence. Most (55.5%) patients had HIV CRF63&#95;02A6, followed by sub-subtype A6 (39.2%). This study revealed poor ART adherence as a main factor driving ART failure among PLWH in the Siberian region. DRM was detected in over 60% of PLWH experiencing ART failure. The current results highlight an urgent need for the introduction of special programs focusing on ART adherence improvement., (Copyright © 2023 Elsevier Ltd and International Society of Antimicrobial Chemotherapy. All rights reserved.)

Objectives: Antiretroviral (ARV) drugs have played a vital role in controlling the HIV-1 epidemic; however, some challenges remain. ARV drugs vary in their ability to control HIV infection, displaying differences in treatment-limiting factors and genetic barriers to resistance. The current report assesses the prevalence of HIV-1 drug resistance mutations (DRMs) among patients who failed first-line antiretroviral therapy (ART) and evaluates the genetic barrier of different regimens., Methods: The study cohort (n = 271) included HIV-infected individuals who visited the Novosibirsk, Russia, HIV/AIDS clinic in 2018-2022. All patients received first-line ART prior to virological failure. Sociodemographic and HIV-related data were collected from medical records and self-reported questionnaires. HIV-1 pol gene sequences were generated, and the presence of HIV-1 DRM was assessed. The genetic barrier to resistance was assessed by combining treatment regimen and adherence data., Results: Nonoptimal ART adherence was identified in 48.3% of patients and correlated with male sex, PWID, unemployment, and rural area residence. Most of the patients with high-level adherence were identified among those who were on TDF+3TC+DTG. HIV-1 DRMs were identified in 54.6% of the patients. The analysis of HIV-1 DRM, ART regimen, and adherence data classified TDF+3TC+DTG and TDF+3TC+LPV/r as treatment regimens with a high genetic barrier, whereas EFV-containing ART was classified as a regimen with a low genetic barrier., Conclusions: The current study delivers results on the efficacy of HIV-1 ART and treatment adherence in real-world practice settings. This report suggests that ART regimens with a high genetic barrier to resistance combined with improved treatment adherence may reduce the transmission of HIV-1 resistant variants., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

HIV-1 epidemic in Russia is one of the fastest growing in the world reaching 1.14 million people living with HIV-1 (PLWH) in 2021. Since mid-1990s, the HIV-1 epidemic in Russia has started to grow substantially due to the multiple HIV-1 outbreaks among persons who inject drugs (PWID) leading to expansion of the HIV-1 sub-subtype A6 (former Soviet Union (FSU) subtype A). In 2006, a local HIV-1 sub-epidemic caused by the distribution of novel genetic lineage CRF63&#95;02A6 was identified in Siberia. In this study, we used a comprehensive dataset of CRF63&#95;02A6 pol gene sequences to investigate the spatiotemporal dynamic of the HIV-1 CRF63&#95;02A6 sub-epidemic. This study includes all the available CRF63&#95;02A6 HIV-1 pol gene sequences from Los Alamos National Laboratory (LANL) HIV Sequence Database. The HIV-1 subtypes of those sequences were conferred using phylogenetic analysis, and two automated HIV-1 subtyping tools Stanford HIVdb Program and COMET. Ancestral state reconstruction and origin date were estimated using Nextstrain. Evolutionary rate and phylodynamic analysis were estimated using BEAST v 1.10.4. CRF63&#95;02A6 was assigned for 872 pol gene sequences using phylogenetic analysis approach. Predominant number (n = 832; 95.4%) of those sequences were from Russia; the remaining 40 (4.6%) sequences were from countries of Central Asia. Out of 872 CRF63&#95;02A6 sequences, the corresponding genetic variant was assigned for 75.7 and 79.8% of sequences by Stanford and COMET subtyping tools, respectively. Dated phylogenetic analysis of the CRF63&#95;02A6 sequences showed that the virus most likely originated in Novosibirsk, Russia, in 2005. Over the last two decades CRF63&#95;02A6 has been widely distributed across Russia and has been sporadically detected in countries of Central Asia. Introduction of new genetic variant into mature sub-subtype A6 and CRF02&#95;AG FSU epidemics could promote the increase of viral genetic diversity and emergence of new recombinant forms. Further HIV-1 studies are needed due to a continuing rapid virus distribution. Also, the implementation of HIV-1 prevention programs is required to reduce HIV-1 transmission. This study also highlights the discrepancies in HIV-1 subtyping approaches. The reference lists of HIV-1 sequences implemented in widely used HIV-1 automated subtyping tools need to be updated to provide reliable results., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Sivay, Maksimenko, Osipova, Nefedova, Gashnikova, Zyryanova, Ekushov, Totmenin, Nalimova, Ivlev, Kapustin, Pozdnyakova, Skudarnov, Ostapova, Yaschenko, Nazarova, Chernov, Ismailova, Maksutov and Gashnikova.)

The aim of this study was to determine the frequency and the clinical and laboratory features of acute viral gastroenteritis in adult patients the residents of Novosibirsk., Materials and Methods: A total of 363 patients aged 16 to 82 years hospitalized with a diagnosis of"acure gastroenteriris" in the winter-spring season 2016 and with no evidence of immunosuppression were examined. In addition to generally accepted diagnostic techniques faeces were investigated using polymerase chain reaction for the detection and differentiation group A and C rotaviruses (HRVA and HRVC), I norovirus genogroup II (HNoV Gil) and astroviruses (HAstV) with applying the original set of specific primers., Results: Viral etiology has been set at 19,6% of patients with acute gastroenteritis. Noroviruses were predominant (10,2%), the proportion of rotavirus was 7,7%, astroviruses - 1,7%. The proportion of viral gastroenteritis was higher in the winter months and decreased in spring months. The moderate form of the disease dominated (99.4%). No significant differences in clinical symptoms, blood count and coprogram parameters were revealed in patients with different etiology of viral gastroenteritis, except the higher frequency of lymphocytosis in norovirus infection (p < 0,05)., Conclusion: Set frequency of viral gastroenteritis in adults, no significant differences in the clinical picture of viral gastroenteritis of various etiologies determine the necessity of elaboration and introduction into clinical practice of universal test systems for the detection of common viral pathogens.

The efficiency of one-step and multi-step protocols of DNA isolation from lysed sputum samples containing the Mycobacterium tuberculosis complex has been compared. DNA was isolated using spin-cartridges containing a special silica-based sorbent modified with fluoroplast and polyaniline, or using an automated isolation system. One-step isolation using the obtained sorbent has been shown to ensure a significantly lower DNA loss and higher sensitivity in the PCR detection of Mycobacterium tuberculosis as compared to a system based on sorption and desorption of nucleic acids during the isolation.

Aims: To demonstrate the effectiveness of application of the adsorbent successively modified with nano-layers of fluoroplast and polyaniline for one-step isolation of DNA of hepatitis B virus and transfusion-transmitted virus from human serum., Materials & Methods: The technique is based on the application of the spin-cartridges containing porous adsorbent for one-step viral DNA isolation from serum followed by polymerase chain reaction., Results: The developed adsorbent was shown to be effective for one-step isolation of viral DNA from serum samples for polymerase chain reaction diagnostics., Conclusion: The effectiveness of the developed adsorbent application for isolation of viral DNA from serum for polymerase chain reaction diagnostics was confirmed in comparison with standard methods. Thus, the facile sample preparation method of viral DNA isolation was elaborated.

Aims: A new approach for the preparation of adsorbents for one-step isolation/purification of DNA from different samples (e.g., bacterial lysates, smears and blood) has been developed., Materials & Methods: The technique is based on the use of porous silica preliminary treated with ozone followed by grafting of polymer nanolayers on the activated carrier surface. The chemical nature of active centers, their stability and conditions for the use of the activated support as heterogeneous initiator of different macromolecular reactions were studied., Results: The adsorbents modified with thin (~3.0-7.5 nm) polytetrafluoroethylene and polyaniline layers were prepared and characterized. Sorption properties of the obtained adsorbents were demonstrated on examples of express (2-5 min) one-step DNA isolation for direct use in PCR diagnostics., Conclusion: The effectiveness of the developed adsorbents used for DNA isolation and purification was confirmed in comparison with the standard methods. Thus, a facile (alternative in relation to irradiation postpolymerization or standard oxidative polymerization techniques) manufacturing method of the materials for nucleic acids sample preparation was developed.

A composite sorbent based on porous glass beads modified with thin polyaniline coating was prepared by precipitating aniline polymerization in the presence of carrier particles. It was shown that the modification ensures the uniform coating of the inner surface of the carrier pores with the polymer layer approximately 70 A thick. It was shown that the resulting material retains the initial porosity of the carrier and is selective in the separation of nucleic acids and proteins. The polyaniline-coated sorbents were shown to be efficient for both the preparative DNA isolation from bacterial lysates and for analytical purposes, in particular, for studying DNA fragmentation during apoptosis proceeding under UV irradiation of cell lysates of colon carcinoma. The morphological and chromatographic characteristics of the new sorbent were demonstrated to be similar to those of the polyfluorobutadiene sorbent.

Composite fluoropolymer-containing sorbents based on porous silicas were synthesized for the isolation and purification of biopolymers under nondenaturing conditions. Examples of the application of these sorbents in the separation of various mixtures of peptides and proteins and purification of nucleic acids from various sources (plasmid DNA and DNA from nucleated human blood cells) using the cartridge, column, and batch (sorption in a stirred volume) methods are presented. It was shown that the sorbents can be used in laboratory practice because they are selective to nucleic acids (DNA and RNA) and proteins. These materials combine the mechanical properties of the inorganic matrix with the specific sorption properties of the polymer phase and exhibit enhanced stability to alkaline hydrolysis. Alternative methods of preparing sorbents containing polytetrafluoroethylene, polytrifluorostyrene, and polyfluorobutadiene are described. By the example of polyfluorobutadiene-containing sorbents, a completely new method for obtaining fluorinated polymer phases was developed: the polymer phase was preliminarily formed on the surface of porous disperse carriers and was fluorinated with xenon difluoride.