The highly conserved human leukocyte antigen-A2(HLA-A2)-restricted epitope NS3-1073 represents a promising candidate for a therapeutic vaccine against hepatitis C virus(HCV).In this study,we engineered a set of fusion...The highly conserved human leukocyte antigen-A2(HLA-A2)-restricted epitope NS3-1073 represents a promising candidate for a therapeutic vaccine against hepatitis C virus(HCV).In this study,we engineered a set of fusion proteins based on the artificial self-assembling peptide(SAP),which were expressed in Escherichia coli and spontaneously self-assembled into nanosized particles displaying HCV epitopes,including NS3-1073.To enhance immunogenicity,we incorporated the T helper epitope PADRE into the construct.Alpha-helical linkers were introduced between SAP and the epitopes to facilitate proper protein folding.Notably,a helical linker with a high supercoiling propensity enabled soluble expression of the fusion protein containing both the NS3-1073 and PADRE epitopes,allowing purification of the in vivo-formed nanoparticles by metal affinity chromatography.Human dendritic cells derived from peripheral blood monocytes showed robust activation in response to the fusion proteins and preferentially stimulated T lymphocytes toward a Th1-biased immune response.In mice,immunization with nanoparticles carrying NS3-1073 induced splenocyte proliferation in response to in vitro stimulation with a mixture of NS3 peptides.These results demonstrate that recombinant nanoparticle-based carriers presenting the NS3-1073 epitope can be produced in bacterial systems and hold strong potential as a foundation for a therapeutic HCV vaccine.展开更多
Isolates of Newcastle disease virus (NDV) from deceased wild and domestic pigeons in Kazakhstan were obtained from the Almaty region during 2005 and were genotypically analyzed by using reverse transcription polymeras...Isolates of Newcastle disease virus (NDV) from deceased wild and domestic pigeons in Kazakhstan were obtained from the Almaty region during 2005 and were genotypically analyzed by using reverse transcription polymerase chain reaction (RT-PCR) with primers specific to the viral fusion (F) protein gene. Part of the amplified F protein DNA product (nucleotide sequence 47-422) and the deduced amino acid sequences were compared phylogenetically with those from strains previously reported in other geographic regions. Phylogenetic analysis indicated that the Kazakhstanian pigeon paramyxovirus type 1 (PPMV-1) isolates belong to genotype VI or 4bii. To our knowledge, this is the first reported VI isolates that possess the sequences of 112 GKRQKR116* F117 within the F0 protein. The information is fundamental to improving the efficiency of control strategies and vaccine development for NDV.展开更多
Although the important role of the non-structural (NS1 and NEP) gene of influenza A in virulence of the virus is well established, our knowledge about the extent of variation in the NS gene pool of influenza A viruses...Although the important role of the non-structural (NS1 and NEP) gene of influenza A in virulence of the virus is well established, our knowledge about the extent of variation in the NS gene pool of influenza A viruses in their natural reservoirs in Kazakhstan is incomplete. 17 influenza A viruses of different subtypes were studied in this paper. Seven types of haemagglutinin and five different neuraminidase subtypes in eight combinations were found among the isolated viruses. A comparison of nucleotide sequences of isolated viruses revealed a substantial number of silent mutations, which results in high degree of homology in amino acid sequences. By phylogenetic analysis it was shown that two distinct gene pools, corresponding to both NS allele A with 5 Clades and B, were present at the same time in Kazakhstan. The degree of variation within the alleles was very low. In our study allele A viruses had a maximum of 5% amino acid divergence in Clade while allele B viruses had only 4% amino acid divergence.展开更多
HIV-1 matrix protein (MA) is a multifunctional structural protein localized on N terminus of Gag precursor p55. MA participates in HIV-1 assembly as membranotropic part of Gag precursor as well as an individual protei...HIV-1 matrix protein (MA) is a multifunctional structural protein localized on N terminus of Gag precursor p55. MA participates in HIV-1 assembly as membranotropic part of Gag precursor as well as an individual protein spliced from Gag early in infection. MA is found in the nuclei of infected cells and in plasma membrane, the site of virus assembly, in association with viral genome RNA. MA mutated variant M4 which contains two changed amino acids in N-terminal regions is also associated with viral RNA, but it is localized in the nuclear and cytoskeleton fractions but not in the plasma membrane suggesting that the mutant is deprived of membranotropic signal and “sticks” in the nuclei an d cytoskeleton, its previous location sites. These data allow suggesting that MA involved into transmission of viral RNA is transported to plasma membrane by cytoskeleton.展开更多
Influenza viruses and coronaviruses have linear single-stranded RNA genomes with negative and positive sense polarities and genes encoded in viral genomes are expressed in these viruses as positive and negative genes,...Influenza viruses and coronaviruses have linear single-stranded RNA genomes with negative and positive sense polarities and genes encoded in viral genomes are expressed in these viruses as positive and negative genes,respectively.Here we consider a novel gene identified in viral genomes in opposite direction,as positive in influenza and negative in coronaviruses,suggesting an ambisense genome strategy for both virus families.Noteworthy,the identified novel genes colocolized in the same RNA regions of viral genomes,where the previously known opposite genes are encoded,a so-called ambisense stacking architecture of genes in virus genome.It seems likely,that ambisense gene stacking in influenza and coronavirus families significantly increases genetic potential and virus diversity to extend virus-host adaptation pathways in nature.These data imply that ambisense viruses may have a multivirion mechanism,like"a dark side of the Moon",allowing production of the heterogeneous population of virions expressed through positive and negative sense genome strategies.展开更多
The efficiency of devices for bioelectronic applications,including cell and tissue stimulation,is heavily dependent on the scale and the performance level.With miniaturization of stimulation electrodes,achieving a suf...The efficiency of devices for bioelectronic applications,including cell and tissue stimulation,is heavily dependent on the scale and the performance level.With miniaturization of stimulation electrodes,achieving a sufficiently high current pulse to elicit action potentials becomes an issue.Herein we report on our approach of vertically stacking organic p-n junctions to create highlyefficient multilayered organic semiconductor(MOS)photostimulation device.A tandem arrangement substantially increases the photovoltage and charge density without sacrificing lateral area,while not exceeding 200-500 nm of thickness.These devices generate 4 times higher voltages and at least double the charge densities over single p-n junction devices,which allow using lower light intensities for stimulation.MOS devices show an outstanding stability in the electrolyte that is extremely important for forthcoming in vivo experiments.Finally,we have validated MOS devices performance by photostimulating fibroblasts and neuroblasts,and found that using tandem devices leads to more effective action potential generation.As a result,we obtained up to 4 times enhanced effect in cell growth density using 3 p-n layered devices.These results corroborate the conclusion that MOS technology not only can achieve parity with state-of-the-art silicon devices,but also can exceed them in miniaturization and performance for biomedical applications.展开更多
Birds at wholesale or retailmarkets which selling live poultry are recognized as important places where some reassortation processes might happen to form new variants,such as reported H7N9,H5N6 and H4N2[1,2].In this c...Birds at wholesale or retailmarkets which selling live poultry are recognized as important places where some reassortation processes might happen to form new variants,such as reported H7N9,H5N6 and H4N2[1,2].In this case,workers in the poultry business are under the high risk of being directly infected by influenza viruses.The recent study presented by Quan et al.展开更多
基金supported by the Russian Science Foundation(Grant No.24-25-20087 to V.K.)。
文摘The highly conserved human leukocyte antigen-A2(HLA-A2)-restricted epitope NS3-1073 represents a promising candidate for a therapeutic vaccine against hepatitis C virus(HCV).In this study,we engineered a set of fusion proteins based on the artificial self-assembling peptide(SAP),which were expressed in Escherichia coli and spontaneously self-assembled into nanosized particles displaying HCV epitopes,including NS3-1073.To enhance immunogenicity,we incorporated the T helper epitope PADRE into the construct.Alpha-helical linkers were introduced between SAP and the epitopes to facilitate proper protein folding.Notably,a helical linker with a high supercoiling propensity enabled soluble expression of the fusion protein containing both the NS3-1073 and PADRE epitopes,allowing purification of the in vivo-formed nanoparticles by metal affinity chromatography.Human dendritic cells derived from peripheral blood monocytes showed robust activation in response to the fusion proteins and preferentially stimulated T lymphocytes toward a Th1-biased immune response.In mice,immunization with nanoparticles carrying NS3-1073 induced splenocyte proliferation in response to in vitro stimulation with a mixture of NS3 peptides.These results demonstrate that recombinant nanoparticle-based carriers presenting the NS3-1073 epitope can be produced in bacterial systems and hold strong potential as a foundation for a therapeutic HCV vaccine.
基金USDA-ISTC partner project(K-747p,Institute of Microbiology and Virology funding,and USDA CRIS(6612-32000-038-00D)
文摘Isolates of Newcastle disease virus (NDV) from deceased wild and domestic pigeons in Kazakhstan were obtained from the Almaty region during 2005 and were genotypically analyzed by using reverse transcription polymerase chain reaction (RT-PCR) with primers specific to the viral fusion (F) protein gene. Part of the amplified F protein DNA product (nucleotide sequence 47-422) and the deduced amino acid sequences were compared phylogenetically with those from strains previously reported in other geographic regions. Phylogenetic analysis indicated that the Kazakhstanian pigeon paramyxovirus type 1 (PPMV-1) isolates belong to genotype VI or 4bii. To our knowledge, this is the first reported VI isolates that possess the sequences of 112 GKRQKR116* F117 within the F0 protein. The information is fundamental to improving the efficiency of control strategies and vaccine development for NDV.
基金USDA-ISTC(The International Science and Technology Center) partner project #К-747рInstituteof Microbiology and Virology project #0103KZО0126 and #0106KZ00581
文摘Although the important role of the non-structural (NS1 and NEP) gene of influenza A in virulence of the virus is well established, our knowledge about the extent of variation in the NS gene pool of influenza A viruses in their natural reservoirs in Kazakhstan is incomplete. 17 influenza A viruses of different subtypes were studied in this paper. Seven types of haemagglutinin and five different neuraminidase subtypes in eight combinations were found among the isolated viruses. A comparison of nucleotide sequences of isolated viruses revealed a substantial number of silent mutations, which results in high degree of homology in amino acid sequences. By phylogenetic analysis it was shown that two distinct gene pools, corresponding to both NS allele A with 5 Clades and B, were present at the same time in Kazakhstan. The degree of variation within the alleles was very low. In our study allele A viruses had a maximum of 5% amino acid divergence in Clade while allele B viruses had only 4% amino acid divergence.
文摘HIV-1 matrix protein (MA) is a multifunctional structural protein localized on N terminus of Gag precursor p55. MA participates in HIV-1 assembly as membranotropic part of Gag precursor as well as an individual protein spliced from Gag early in infection. MA is found in the nuclei of infected cells and in plasma membrane, the site of virus assembly, in association with viral genome RNA. MA mutated variant M4 which contains two changed amino acids in N-terminal regions is also associated with viral RNA, but it is localized in the nuclear and cytoskeleton fractions but not in the plasma membrane suggesting that the mutant is deprived of membranotropic signal and “sticks” in the nuclei an d cytoskeleton, its previous location sites. These data allow suggesting that MA involved into transmission of viral RNA is transported to plasma membrane by cytoskeleton.
文摘Influenza viruses and coronaviruses have linear single-stranded RNA genomes with negative and positive sense polarities and genes encoded in viral genomes are expressed in these viruses as positive and negative genes,respectively.Here we consider a novel gene identified in viral genomes in opposite direction,as positive in influenza and negative in coronaviruses,suggesting an ambisense genome strategy for both virus families.Noteworthy,the identified novel genes colocolized in the same RNA regions of viral genomes,where the previously known opposite genes are encoded,a so-called ambisense stacking architecture of genes in virus genome.It seems likely,that ambisense gene stacking in influenza and coronavirus families significantly increases genetic potential and virus diversity to extend virus-host adaptation pathways in nature.These data imply that ambisense viruses may have a multivirion mechanism,like"a dark side of the Moon",allowing production of the heterogeneous population of virions expressed through positive and negative sense genome strategies.
基金funded by the Ministry of Science and Higher Education of the Russian Federation(No.075-15-2021-596).
文摘The efficiency of devices for bioelectronic applications,including cell and tissue stimulation,is heavily dependent on the scale and the performance level.With miniaturization of stimulation electrodes,achieving a sufficiently high current pulse to elicit action potentials becomes an issue.Herein we report on our approach of vertically stacking organic p-n junctions to create highlyefficient multilayered organic semiconductor(MOS)photostimulation device.A tandem arrangement substantially increases the photovoltage and charge density without sacrificing lateral area,while not exceeding 200-500 nm of thickness.These devices generate 4 times higher voltages and at least double the charge densities over single p-n junction devices,which allow using lower light intensities for stimulation.MOS devices show an outstanding stability in the electrolyte that is extremely important for forthcoming in vivo experiments.Finally,we have validated MOS devices performance by photostimulating fibroblasts and neuroblasts,and found that using tandem devices leads to more effective action potential generation.As a result,we obtained up to 4 times enhanced effect in cell growth density using 3 p-n layered devices.These results corroborate the conclusion that MOS technology not only can achieve parity with state-of-the-art silicon devices,but also can exceed them in miniaturization and performance for biomedical applications.
文摘Birds at wholesale or retailmarkets which selling live poultry are recognized as important places where some reassortation processes might happen to form new variants,such as reported H7N9,H5N6 and H4N2[1,2].In this case,workers in the poultry business are under the high risk of being directly infected by influenza viruses.The recent study presented by Quan et al.
