Considering the wide spread of known and new pathogens of acute respiratory viral infections (ARVI), it is important to search for drugs that have a wide spectrum of antiviral activity and additionally the ability to act pathogenetically Equally important is the confirmation of therapeutic activity in drugs known for their effectiveness and safety In order to monitor the susceptibility of ARVI pathogens to the antiviral drug Influcid, a series of in vitro studies was performed Materials and methods: classical virological controlled studies were performed using current strains of human influenza virus (A/Moscow/225/2019 (H1N1)pdm09 and B/Moscow/17/2019) and MDCK cell culture Antiviral, cytoprotective (against viruses, antiviral and antipyretic drugs) properties of the Inflicid were evaluated by changing the logarithm of the tissue cytopathic infecting dose, which causes 50% of cells to be damaged (lgTCID50), as well as by changing the optical density when using tetrazolium dyes Results and discussion: Influcid dose-dependently suppressed the reproduction of epidemic strains A/Moscow/225/2019(H1N1)pdm09 and B/Moscow/17/2019 by 50% at concentrations of 0, 34 and 0, 69-0, 86% vol , respectively Also, the Influcid reduced the reproduction of human influenza virus strains: A/New Caledonia/20/99(H1N1), A/Victoria/35/72, Wisconsin/67/05(H3N2) and B/Malaysia/2506/04, reducing the titers by 0, 5-2, 5 lgTCID50 and Dengue virus DENV-2/RUS/TH-Novosibirsk02/2012 Influcid exerted a cytoprotective effect on cells infected with A/IIV-Moscow/01/2009 (H1N1)pdm09, increasing the proportion of surviving cells by 4-52, 0% For cells infected with the pandemic strain A/California/07/09(H1N1)pdm09, there was a significant decrease in the cytopathic effect of the virus up to full protection with the «preventive» scheme of the drug application In addition, the Influcid protected cells from the action of herpes viruses types I and II, type III adenovirus, parainfluenza virus, seasonal coronavirus and respiratory syncytial virus, toxic doses of umifenovir and rimantadine, and did not increase the cytotoxicity of acetaminophen and ibuprofen Conclusion: having a combination of confirmed antiviral activity against current epidemic virus strains, cytoprotective action, the ability to exert a pathogenetic (immunomodulatory and anti-inflammatory) effect, as well as a favorable safety profile, Influcid may be a relevant and promising drug in the arsenal of medical practitioners

Influenza virus has two major structural modules, an external lipid envelope and an internal ribonucleocapsid containing the genomic RNA in the form of the ribonucleoprotein (RNP) complex, both of which are interlinked by the matrix protein M1. Here we studied M1-RNP cohesion within virus exposed to acidic pH in vitro. The effect of acidification was dependent on the cleavage of the surface glycoprotein HA. Acidic pH caused a loss of intravirion RNP-M1 cohesion and activated RNP polymerase activity in virus with cleaved HA (HA1/2) but not in the uncleaved (HA0) virus. The in vitro acidified HA1/2 virus rapidly lost infectivity whereas the HA0 one retained infectivity, following activation by trypsin, suggesting that premature activation and release of the RNP is detrimental to viral infectivity. Rimantadine, an inhibitor of the M2 ion channel, was found to protect the HA1/2 virus interior against acidic disintegration, confirming that M2-dependent proton translocation is essential for the intravirion RNP release and suggesting that the M2 ion channel is only active in virions with cleaved HA. Acidic treatment of both HA0 and HA1/2 influenza viruses induces formation of spikeless bleb-like protrusion of ~25nm in diameter on the surface of the virion, though only the HA1/2 virus was permeable to protons and permitted RNP release. It is likely that this bleb corresponds to the M2-enriched and M1-depleted focus arising from pinching off of the virus during the completion of budding. Cooperatively, the data suggest that the influenza virus has an asymmetric structure where the M1-mediated organization of the RNP inside the virion is a prerequisite for infectious entry into target cell.

Dynamic light scattering method or laser correlation spectroscopy was applied to evaluation of the size of viruses. We measured correlation functions of the light scattered by human immunodeficiency viruses (HIV) and hepatitis A viruses (HAV) and found that size of HIV-1 (subtype A and B) and HAV virions were 104 nm and 28 nm, respectively. Comparison of these findings with electron microscopy data for fixed samples of the same viruses showed good agreement of the results.

Vesicles on the virion surface, which continued the lipoprotein membrane but had no spikes of virus glycoproteins hemagglutinin (HA) and neuraminidase (NA), were detected. These vesicles and virus particles were 18±7 and 103±12 nm in diameter, respectively, and, as a rule, one vesicle was found per virion. The locus of the external protrusion in the virion presumably corresponded to the site of virus budding during assembly in infected cell free from HA and NA spikes outside and M1 matrix protein inside, but enriched with ionic channel protein M2. Particles with vesicles constituted ~3-10% of the virus population produced in MDCK-H culture and containing uncleaved HA0 hemagglutinin. The content of vesicular virions increased slightly after trypsin cleavage HA0→HA1+HA2 and reached 10-15%. Exposure of the virus in acid medium (pH 4.3) led to a drastic increase of vesicular virions – to 60-80% for HA0 and HA1+HA2 virus. This was paralleled by changes in contrast permeability (for phosphotungstic acid). HA0 virions remained contrast-impermeable, while HA1+HA2 particles let the contrast in through the vesicles detected on the surface and were rapidly destroyed after incubation in acid medium. Hence, cleavage of the surface glycoprotein HA0 into HA1 and HA2 stimulated the acid-dependent permeability of the lipid membrane and led to attenuation of the ribonucleoprotein and protein matrix M1 contacts inside the virion.

The comparative examination of the interaction of the influenza A and B viruses and fragments of DNA with the carbon nanotubes - composites of polyaniline (PANI) nanotubes and granules containing Ag and without Ag was performed. The increased absorption of the allantois viruses and DNA was demonstrated in composites with Ag. The influence of temperature in the range of 4-36° C was not found to be essential. The intensive absorption took place within the first 15 min of the contact with the sorbents. in total, the properties of the composites of PANI nanotubes + Ag 30% are the most promising for the influenza viruses and DNA absorption in water solutions.

The Velmi thermal-laser therapeutic system for treatment of genital human papillomavirus (HPV) infection using photoactivated drug is described. The results of clinical use of this method are discussed. The efficacy of HPV elimination in the uterine cervix was supported by conventional clinical methods of HPV diagnosis (including electron microscopy).

Получены полианилин и его композиты с углеродными нанотрубами или с наночастицами серебра, которые охарактеризованы методами электронной микроскопии, спектроскопии, кондуктометрии и БЭТ-анализа. Изучена in vitro и in vivo токсичность материалов, после чего они использованы в качестве сорбентов вирусов гриппа А и В. Показано, что наилучшей сорбционной способностью по отношению ко всем штаммам вирусов обладает композит полианилина с серебром, который обеспечивает практически полное удаление вирусов из водного раствора.

Polyaniline and its composites with carbon nanotubes or silver nanoparticles are prepared and characterized via electron microscopy, spectroscopy, conductometry, and BET analysis. The in vitro and in vivo toxicities of the materials are studied, and then the materials are used as sorbents of the influenza A and B viruses. It is shown that the best sorption capacity with respect to all of the virus strains is exhibited by the polyaniline composite with silver, which provides for almost complete removal of viruses from aqueous solutions.

The perspectives of the application of modern materials contained nanodiamonds (ND) are considered in this study. The interaction between detonation particulate ND, soot and influenza A and B viruses, fragments of cDNA were analyzed at the normal conditions. It was shown that these sorbents can interact with the following viruses: reference epidemic strains of influenza A(H1N1), A(H1N1) v, A(H3N2) and B viruses circulated in the word in 2000-2010. The allantoises, concentrated viruses, cDNA can be absorbed by ND sorbents and getting removed from water solutions within 20 min. ND sorbents can be used for the preparation of antivirus filters for water solution and for future diagnostic systems in virology.

The paper presents data on the sorption of influenza A(H1N1), A(H1N1)v, A(H3N2) viruses, cDNA of A(H1N1)v and B viruses on nanodiamonds and furnace charge. The sorption of viruses occurred in different solutions at 4-37° C during 10-20 min. The rate of sorption varied with the concentration of a sorbent in the solution and its structure, but did not with the antigenic formula of viruses or temperature. The sorption capacity of furnace charge towards influenza A and B viruses was higher than that of nanodiamonds. Nonviral proteins (bovine serum albumin and influenza virus antibodies) were found to be bound by both sorbents. Viral desorption did not take place in physiological solution at 4 and 22° C for 48 hours.

Virus particles are typically consisted of several hundreds to thousands of protein molecules, which self-assembled to form hollow scaffold containing the viral nucleic acid (RNA or DNA). They ranged in sizes from ~ 10 nm to over a micron and can be found in variety of distinctive shapes (icosahedrons, spheres, rodes). As an emerging and important nanocarrier platform, viruses and virus-like particles (VLP) offer the great advantages of morphological uniformity, biocompatibility and polyfunctionality. In recent years, viruses and VLPs are tailorable at the genetic level for application as reagents, catalysts and scaffolds for chemical reactions, effective vectors for targeted drug delivery and suitable platform for candidate vaccine constructions. This review describes approaches to use viruses in different spheres of nanotechnology and nanomedicine for the next generation therapeutics and imaging devices, as nanocomposites with chemical compounds and metals for new diagnostic methods and even in electronics. Several examples in nanobiomedicine researches at D.I. Ivanovsky Institute of Virology (new diagnostic biochips for influenza A subtypes differentiation, latex-agglutination method for HCV antibodies detection, atomic force microscopy in clinics, TMV recombinants expressing influenza HA antigenic epitopes, polyanylin and nanodiamonds as effective biosorbents) are mentioned as promissive for future development.

This paper presents the data on sorption efficiency of nanodiamonds, detonation soot and its complexes with polyaniline for nano size biological objects: fragments of cDNA and epidemic influenza viruses, including A(H1N1, H3N2) and B strains, isolated in 1999-2010, pandemic strain A(H1N1)pdm09, influenza avian A(H5N2) viruses and reassortants A(H5N2) and A(H5N1). As the result of the sorption on detonation nanodiamonds (DND), detonation soot and their complexes with polyaniline, hemagglutination (HA) titers of virus solutions decreased by ≤ 4000 times for concentrated viruses, suspended in saline and by ≤8 times for allantoic viruses. The detonation soot was approximately 2.5 times more effective than DND. Detonation nanodiamond materials proved to be efficient sorbents for cDNA fragments of >190 bp size. No desorption of viruses from nanodiamonds and soot into saline was revealed (observation time 48 h). DND and soot at concentrations ≤ 1 mg/ml were not toxic for MDCK cells. A moderate increase of monocytes was registrated in blue rats after intra-abdominal immunization with DND+ virus complexes.

Elimination of types 16 and 18 human papilloma virus from the surface of cervix uteri for secondary prevention of cervical cancer was evaluated. The method is protected by patent of invention of the Russian Federation. Infrared laser therapy of cervix uteri was carried out in patients with precancer diseases of cervix uteri at Department of Gynecology of Municipal Clinical Hospital No. 52 (Moscow). Papillomavirus infection was eliminated using a Russian diode laser (λ = 1.06 μ, radiation power 10 W) with a collimating headpiece using carbon die at a distance of 10-12 cm from the exposed surface. The treatment resulted in a high percentage of elimination of types 16 and 18 oncogenic virus 4-6 weeks and during delayed periods after exposure.

The paper gives data on the sorption of influenza virus pandemic strain A/IIV-Moscow/01/2009 (H1N1)swl, avian influenza viruses with A/H5 and A/H7 hemagglutinin, poliomyelitis virus, and T4-D bacteriophage on polyaniline sorbents, carbon nanotubes, and their based nanocomposites. The sorption of viruses occurred in different solutions at 4-37 degrees C during 15 min or more. The rate of viral sorption depended on the structure of sorbents.

Complexes between influenza viruses proteins and antibodies to them create the precipitation lines in polyaniline (PM) films which were detected by polarization optical microscopy after coating the film from the top the eutectic mixture of nematic liquid crystals MBBA:EBBA. Polyaniline and it's composites interact as sorbents with influenza human and bird viruses apart from antigenic properties of surface proteins as well as with bacteriophage T4-D. The sorption of virions occurred in different solutions in the temperature range (8-25 degrees C) during 15 - 60 min. After contact with sorbent the HA titers of influenza virus in solutions were decreased in 4 -2048 times. The immobilized influenza viruses were able to interact with homological antibodies from immune sera in solution.