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A new in vitro model of Huntington's disease (HD) was developed via a direct reprogramming of dermal fibroblasts from HD patients into striatal neurons. A reprogramming into induced pluripotent stem (iPS) cells is obviated in the case of direct reprogramming, which thus yields neurons that preserve the epigenetic information inherent in cells of a particular donor and, consequently, the age-associated disease phenotype. A main histopathological feature of HD was reproduced in the new model; i.e., aggregates of mutant huntingtin accumulated in striatal neurons derived from a patient's fibroblasts. Experiments with cultured neurons obtained via direct reprogramming make it possible to individually assess the progression of neuropathology and to implement a personalized approach to choosing the treatment strategy and drugs for therapy. The in vitro model of HD can be used in preclinical drug studies.
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Enfermedad de Huntington , Células Madre Pluripotentes Inducidas , Humanos , Animales , Enfermedad de Huntington/genética , Enfermedad de Huntington/patología , Neuronas , Cuerpo Estriado/patología , Fibroblastos , Células Madre Pluripotentes Inducidas/patología , Modelos Animales de EnfermedadRESUMEN
The transcription factor NF-κB is a key factor in the activation of immune responses; it is in turn activated by pattern recognition receptors, such as TLR and NLR receptors. The search for ligands activating innate immunity receptors is an important scientific problem due to the possibility of their use as adjuvants and immunomodulators. In this study the effect of recombinant Pseudomonas aeruginosa OprF proteins and a toxoid (a deletion atoxic form of exotoxin A) on the activation of TLR4, TLR9, NOD1, and NOD2 receptors has been investigated. The study was carried out using free and co-adsorbed on Al(OH)3 P. aeruginosa proteins and eukaryotic cells encoding these receptors and having NF-κB-dependent reporter genes. The enzymes encoded by the reported genes are able to cleave the substrate with the formation of a colored product, the concentration of which indicates the degree of receptor activation. It was found that free and adsorbed forms of the toxoid were able to activate the TLR4 surface receptor for lipopolysaccharide. OprF and the toxoid activated the intracellular NOD1 receptor, but only in the free form. This may be due to the fact that the cell lines used were not able to phagocytize aluminum hydroxide particles with protein adsorbed on them.
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FN-kappa B , Pseudomonas aeruginosa , Células Eucariotas , Receptor Toll-Like 4/genética , Toxoides , Adyuvantes Inmunológicos , Proteínas Recombinantes/genéticaRESUMEN
A recombinant form of pneumolysin from Streptococcus pneumoniae was obtained. By using Vector NTI Advance 11.0 bioinformatic analysis software, specific primers were designed in order to amplify the genome fragment of strain No. 3358 S. pneumoniae serotype 19F containing the nucleotide sequence encoding the full-length pneumolysin protein. A PCR product with a molecular weight corresponding to the nucleotide sequence of the S. pneumoniae genome fragment encoding the full-length pneumolysin was obtained. An expression system for recombinant pneumolysin in E. coli was constructed. Sequencing confirmed the identity of the inserted nucleotide sequence encoding the full-length recombinant pneumolysin synthesized in E. coli M15 strain. Purification of the recombinant protein was performed by affinity chromatography using Ni-Sepharose in 8 M urea buffer solution. Confirmation of the recombinant protein was performed by immunoblotting with monoclonal antibodies to pneumolysin.
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Escherichia coli , Streptococcus pneumoniae , Streptococcus pneumoniae/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Estreptolisinas/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
Chronic kidney disease (CKD) is characterized by high mortality from cardiovascular diseases, the development of which is facilitated by traditional risk factors (typical for the general population) and by nontraditional ones (specific to patients with CKD) as well. These factors include also uremic toxins, for which a causal relationship has been established with specific pathological processes in patients with CKD, comprising the development of vascular dysfunction and accelerated progression of atherosclerosis. Urea has long been considered not as a uremic toxin, but as a marker of metabolic imbalance or dialysis efficiency (Kt/V) in CKD patients. In recent years, more and more publications have appeared on the study of the toxic effects of urea with the development of toxic-uremic complications and the phenotype of premature aging, common in CKD. It was found that an increase in urea levels in uremic syndrome causes damage to the intestinal epithelial barrier with translocation of bacterial toxins into the bloodstream and the development of systemic inflammation, provokes apoptosis of vascular smooth muscle cells, as well as endothelial dysfunction, which directly contributes to the development of cardiovascular complications. The indirect effects of increased urea levels are associated with carbamylation reactions, when isocyanic acid (a product of urea catabolism) changes the structure and function of proteins in the body. Carbamylation of proteins in CKD patients is associated with the development of renal fibrosis, atherosclerosis and anemia. Thus, urea is now regarded as an important negative agent in the pathogenesis of complications in CKD. Studies on a low-protein diet with using ketoanalogues of essential amino acids to minimize the accumulation of urea and other uremic toxins demonstrate the clinical benefit of such an intervention in slowing the progression of CKD and the development of cardiovascular complications.
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Aterosclerosis , Toxinas Bacterianas , Insuficiencia Renal Crónica , Uremia , Humanos , Dieta con Restricción de Proteínas/efectos adversos , Carbamilación de Proteína , Urea , Aminoácidos Esenciales , Tóxinas Urémicas , Insuficiencia Renal Crónica/complicaciones , Proteínas/metabolismo , Toxinas Bacterianas/metabolismo , Aterosclerosis/complicaciones , Uremia/complicaciones , Uremia/metabolismoRESUMEN
Immunogenic and protective activity of recombinant pneumolysin was studied in experiments on male BALB/c mice. The mice were immunized intraperitoneally with recombinant pneumolysin sorbed on Al(OH)3 (200 µg per mouse). In 2 weeks after immunization, the isotypes of antibodies to recombinant pneumolysin in the serum of immunized mice were determined by ELISA. The animals were infected with Streptococcus pneumoniae serotype 3. Immunization with recombinant pneumolysin induced the production of anti-pneumolysin antibodies, mainly of IgG1 subisotype. On day 21 after intraperitoneal infection with S. pneumoniae serotype 3 in a dose of 106 microbial cells, the survival rate of animals immunized with recombinant pneumolysin in a dose of 25 µg/mouse was 67% vs. 0% in the control (p<0.001). Recombinant pneumolysin could be considered as a promising protective antigen for inclusion in the serotype-independent vaccine against S. pneumoniae.
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Anticuerpos Antibacterianos/biosíntesis , Inmunoglobulina G/biosíntesis , Infecciones Neumocócicas/prevención & control , Vacunas Neumococicas/administración & dosificación , Streptococcus pneumoniae/inmunología , Estreptolisinas/administración & dosificación , Adyuvantes Inmunológicos/administración & dosificación , Compuestos de Alumbre/administración & dosificación , Animales , Proteínas Bacterianas/administración & dosificación , Proteínas Bacterianas/biosíntesis , Inmunización/métodos , Inmunogenicidad Vacunal , Inyecciones Intraperitoneales , Masculino , Ratones , Ratones Endogámicos BALB C , Infecciones Neumocócicas/inmunología , Infecciones Neumocócicas/mortalidad , Infecciones Neumocócicas/patología , Vacunas Neumococicas/biosíntesis , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/biosíntesis , Streptococcus pneumoniae/efectos de los fármacos , Streptococcus pneumoniae/patogenicidad , Estreptolisinas/biosíntesis , Análisis de SupervivenciaRESUMEN
Fertilization (gamete fusion followed by zygote formation) is a multistage process. Each stage is mediated by ligand-receptor recognition of gamete interaction molecules. This recognition includes the movement of sperm in the gradient of egg chemoattractants, destruction of the egg envelope by acrosomal proteins, etc. Gametic incompatibility is one of the mechanisms of reproductive isolation. It is based on species-specific molecular interactions that prevent heterospecific fertilization. Although gametic incompatibility may occur in any sexually reproducing organism, it has been studied only in a few model species. Gamete interactions in different taxa involve generally similar processes, but they often employ non-homologous molecules. Gamete recognition proteins evolve rapidly, like immunity proteins, and include many taxon-specific families. In fact, recently appeared proteins particularly contribute to reproductive isolation via gametic incompatibility. Thus, we can assume a multiple, independent origin of this type of reproductive isolation throughout animal evolution. Gametic incompatibility can be achieved at any fertilization stage and entails different consequences at different taxonomic levels and ranges, from complete incompatibility between closely related species to partial incompatibility between distantly related taxa.
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Bone marrow mesenchymal stromal cells are multipotent and can differentiate into cells of various tissues, which determines their high importance for clinical application. We performed an in vitro study of the osteogenic potential of mesenchymal stromal cells cultured on intact polylactide scaffolds or scaffolds modified with collagen I or fibrin. Scanning electron microscopy showed that the cells formed osteogenic nodules or osteogenic nodules on both intact and fibrin-modified polylactide scaffolds. Spectrophotometric detection of alkaline phosphatase activity on days 7 and 11 showed that mesenchymal stromal cell grown on intact polylactide scaffolds and on scaffolds modified with collagen type I or fibrin more intensively synthesized alkaline phosphatase than in the control (culture plastic). This dependence increases in the presence of osteogenic differentiation factors in the medium. After long-term culturing (4 weeks), the presence of calcium deposits detected by alizarin red staining confirmed the osteoinductive properties of intact and protein-modified polylactide scaffolds. These findings suggest that polylactide scaffolds and collagen I increase the osteogenic differentiation potential of mesenchymal stromal cells.
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Poliésteres/química , Andamios del Tejido/química , Fosfatasa Alcalina/metabolismo , Animales , Células de la Médula Ósea/citología , Células de la Médula Ósea/metabolismo , Diferenciación Celular/fisiología , Células Cultivadas , Colágeno Tipo I/metabolismo , Fibrina/metabolismo , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/fisiología , Osteocalcina/metabolismo , Osteogénesis/fisiología , Conejos , Ingeniería de Tejidos/métodosRESUMEN
The formation of pro-/eukaryotic systems is the general biological mechanism of formation and variability of the phenotype of plants, animals, human beings under the influence of external wednesday, i.e. formation of adaptive potency conditions to external wednesday that increases the <
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Epigénesis Genética/inmunología , Tolerancia Inmunológica , Microbiota/inmunología , Animales , Preescolar , Humanos , Lactante , Recién NacidoRESUMEN
AIM: Evaluate antibacterial activity of an experimental mixture of phages, belonging to several well-studied species. MATERIALS AND METHODS: The study was carried out using a group of 55 clinical Pseudomonas aeruginosa strains of various origins,- 4 mono-species mixtures of 32 virulent bacteriophages (species phiKZ-, phiKMV-, phiPBl-, PaP3-like phages) and 2 novel phages, phiMK (species PaK-P2) and phiPerm5. Activity of preparations from mono-species mixtures of bacteriophages ofvarious species were compared with activity of 3 commercial mixtures. Standard methods of study of bacteriophages were used: determination of lytic activity by seeding onto bacterial lawns of P. aeruginosa, restriction analysis of phage DNA for confirmation of their be- longing to certain species. RESULTS: Cumulative activity of 6 mono-species mixtures of virulent phages was shown to be similar to lytic activity of commercial therapeutic mixtures used against P. aeruginosa infections. 54 of 55 strains of clinical isolates of P: aeruginosa showed sensitivity to experimental mixtures composed of mono-species mixtures of bacteriophages. 53 strains were lysed by commercial preparations. Wherein the possibility of accidental inclusion of moderate -bacteriophages in the experimental mixture is excluded. CONCLUSION: A possibility of creation of highly active therapeutic antibacterial preparations against P. aeruginosa using mono-species mixtures of 6 species of lytic bacteriophages is shown Use of such a mixture in therapy of lung infections reduces the risk of emergence of bacterial strains with increased virulence and patho- genicity during prolonged administration.
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Bacteriófagos , Terapia de Fagos , Infecciones por Pseudomonas/terapia , Pseudomonas aeruginosa/virología , HumanosRESUMEN
AIM: Study protective activity of protein-containing antigens of pneumococcus, obtained from serotypes 6B, 10A, 14, 19F, 23F and 36R, against infection with heterologous strains of S. pneumoniae. MATERIALS AND METHODS: S. pneumoniae strains of serotypes 3, 6B, 10A, 14, 19F, 23F and 36R, obtained from the collection of pneumococcus strains of Mechnikov RIVS, were used in the study. Protein-containing antigens of S. pneumoniae were isolated by acetone precipitations of supernatant fraction of culture medium. Protective activity of preparations of protein-containing antigens of pneumococcus as studied in experiments of active protection of BALb/c line mice. RESULTS: The data obtained give evidence, that protein-containing antigens of pneumococcus, isolated from serotypes 6B, 10A, 14, 19F and 23F, effectively protect animals from subsequent infection with a heterologous S. pneumoniae strain of serotype 3 No. 11/56. Protection was noted at a level from 80 to 100% (p ≤ 0.05). Similar protective effect was detected in another experiment in a group of mice, immunized with preparations of protein-containing antigens of pneumococcus, obtained from serotypes 6B and 36R, against infection with a heterologous S. pneumoniae strain of serotype 3 No. 11/56. Protection was noted at a level of 90% (p ≤ 0.05). CONCLUSION: The results of the experiments carried out allow to assume, that the main role in formation of cross-protection in experiments in animals is played by pneumococcus, proteins, that are a part of the studied preparations, and not polysaccharide antigens.
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Antígenos Bacterianos/inmunología , Proteínas Bacterianas/inmunología , Vacunas Neumococicas/inmunología , Neumonía Neumocócica/prevención & control , Streptococcus pneumoniae/inmunología , Vacunación , Animales , Antígenos Bacterianos/administración & dosificación , Antígenos Bacterianos/química , Proteínas Bacterianas/administración & dosificación , Proteínas Bacterianas/química , Protección Cruzada , Medios de Cultivo/química , Humanos , Ratones , Ratones Endogámicos BALB C , Vacunas Neumococicas/administración & dosificación , Vacunas Neumococicas/química , Neumonía Neumocócica/inmunología , Neumonía Neumocócica/microbiología , Serogrupo , Streptococcus pneumoniae/químicaRESUMEN
AIM: Study the effectiveness of the substance and various drug formulations of fullerene-(tris-aminocapronic acid) hydrate (FTAAH onwards) in the model of experimental viral-bacterial pneumonia of mice. MATERIALS AND METHODS: BALB/c mice were infected with influenza virus A/California/04/2009 and subsequently infected with Staphylococcus aureus. The animals were treated after viral infection with the substance and various drug forms of FTAAH, as well as comparative preparations--oseltamivir and arbidol. Therapy effectiveness was evaluated by clinical indicators (survival, lifespan, animal mass decrease reduction), virological (virus titer), microbiological (density of bacteria in lungs) parameters, confirmed by pathomorphological characteristics of lungs. RESULTS: FTAAH therapy in injectable form was effective in the model of a combined viral-bacterial pneumonia of mice by all the studied criteria: treatment increased mice survival, reduced the decrease of their body weight, resulted in a reduction of virus titers and density of bacteria in lungs, that correlated with the data from morphological study and signs of bronchopneumonia resolution in mice. FTAAH therapy in rectal form depended on animal infection schemes, as well as preparation dose, increasing with its increase. CONCLUSION: FTAAH substance is effective in the model of experimental viral-bacterial pneumonia of mice.
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Fulerenos/administración & dosificación , Neumonía Bacteriana/tratamiento farmacológico , Neumonía Viral/tratamiento farmacológico , Animales , Química Farmacéutica , Modelos Animales de Enfermedad , Fulerenos/química , Humanos , Subtipo H1N1 del Virus de la Influenza A/patogenicidad , Pulmón/microbiología , Pulmón/patología , Pulmón/virología , Ratones , Oseltamivir/administración & dosificación , Neumonía Bacteriana/microbiología , Neumonía Bacteriana/patología , Neumonía Bacteriana/virología , Neumonía Viral/microbiología , Neumonía Viral/patología , Neumonía Viral/virología , Staphylococcus aureus/patogenicidadRESUMEN
Salinity is one of the most important abiotic environmental factors affecting marine animals. If salinity deviate from optimum, adaptive mechanisms switch on to maintain organism's physiological activity. In this study, the reaction of the snails Littorina saxatilis from natural habitats and in response to experimental salinity decreasing was analyzed on proteomic level. The isolation of all snails inside their shells and gradually declining mortality was observed under acute experimental salinity decrease (down to 10 per hundred). Proteomic changes were evaluated in the surviving experimental mollusks compared to control individual using differential 2D gel-electrophoresis (DIGE) and subsequent LC-MS/MS-identification of proteins. Approximately 10% of analyzed proteins underwent up- or down regulation during the experiment. Proteins of folding, antioxidant response, intercellular matrix, cell adhesion, cell signaling and metabolic enzymes were identified among them. Proteome changes observed in experimental hypoosmotic stress partially reproduced in the proteomes of mollusks that live in conditions of natural freshening (estuaries). Possible mechanisms involved in the adaptation process of L. saxatilis individuals to hypo-osmotic stress are discussed.
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Regulación de la Expresión Génica , Proteoma/genética , Tolerancia a la Sal/genética , Caracoles/efectos de los fármacos , Cloruro de Sodio/farmacología , Secuencia de Aminoácidos , Animales , Antioxidantes/aislamiento & purificación , Antioxidantes/metabolismo , Cromatografía Liquida , Electroforesis en Gel Bidimensional , Proteínas de la Matriz Extracelular/genética , Proteínas de la Matriz Extracelular/aislamiento & purificación , Proteínas de la Matriz Extracelular/metabolismo , Perfilación de la Expresión Génica , Chaperonas Moleculares/genética , Chaperonas Moleculares/aislamiento & purificación , Chaperonas Moleculares/metabolismo , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Presión Osmótica , Proteoma/aislamiento & purificación , Proteoma/metabolismo , Salinidad , Transducción de Señal , Caracoles/genética , Espectrometría de Masas en TándemRESUMEN
AIM: Study experimental production series of Staphylovac-2 by accumulation of specific IgG and safety. MATERIALS AND METHODS: Experimental production samples of staphylococci vaccines were studied by the accumulation of specific IgG in sera of immunized BALB/c line mice in EIA. Safety was evaluated in tests of acute and chronic toxicity including pathomorphologic and histologic, hematologic and biochemical studies, studies of the effect on central nervous system. RESULTS: A statistically significant (2.6 - 3.0 times) increase of IgG levels in sera of immunized mice compared with control was noted. In the experiments studying acute and chronic toxicity the increase in body mass and mass of internal organs differed from data obtained from control animals at no observation periods. None of the studied methods of safety evaluation showed differences of the studied vaccine series from the control. CONCLUSION: The recommended dose for subcutaneous administration into human of 200 µg is experimentally justified and could be the basis for carrying out clinical studies of staphylococci vaccines in humans.
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Inmunoglobulina G/sangre , Vacunas Estafilocócicas/administración & dosificación , Vacunas Estafilocócicas/inmunología , Animales , Formación de Anticuerpos , Humanos , Inmunización , Ratones , Vacunas Estafilocócicas/efectos adversosRESUMEN
Objectives. To study the risk factors of asymptomatic cerebral infarctions (AI) and their effect on cognitive functions. Material and methods. We examined 114 patients, aged from 53 to 88 years, including 58 patients with AI (group 1), 32 women and 26 men (mean age 66.5±7.3 yeas), and 56 patients with chronic cerebral ischemia (CCI) without a history of stroke and transitory ischemic attacks, (group 2), 36 women and 20 men (mean age 64.2±6.6 yeas). Results. Cardiovascular risk factors (arterial hypertension, diabetes mellitus, ischemic heart disease, atrial fibrillation) were more frequent in group 1. The results of neuropsychological testing revealed worse cognitive performance in patients with AI. Conclusions. The results confirmed the negative effect of AI on cognitive functioning and higher risk of CCI progression.
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AIM: Production, study of properties and evaluation of a possibility to use monoclonal antibodies against Pseudomonas aeruginosa exotoxin A (ETA) for the detection of molecules of recombinant exotoxin A and anatoxin. MATERIALS AND METHODS: Producer hybridomas for monoclonal antibodies against ETA were generated. Recombinant exotoxin A, its atoxic forms, P. aeruginosa anatoxin and P. aeruginosa PA-103 live culture were used for immunization of mice. RESULTS: The experiment of fusion of malignant cell line with immune lymphocytes obtained from a mouse immunized with recombinant ETA turned out to be the most productive. Intensity of interaction of monoclonal antibodies with recombinant atoxic forms of ETA was evaluated in enzyme immunoassay. Protective (toxin-neutralizing) activity of antibodies in cell culture and the ability to detect exotoxin A in latex-agglutination reaction were studied. Antibodies of hybrid culture No. 21 are able to detect molecules of recombinant ETA and anatoxin in the latex-agglutination reaction. CONCLUSION: Use of the antibodies produced for testing toxigenicity of P. aeruginosa strains as well as detection of anatoxin in the technological process of production of prophylaxis preparations based on exotoxin A is expected.
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ADP Ribosa Transferasas/inmunología , Anticuerpos Monoclonales de Origen Murino/inmunología , Toxinas Bacterianas/inmunología , Exotoxinas/inmunología , Pseudomonas aeruginosa/inmunología , Factores de Virulencia/inmunología , ADP Ribosa Transferasas/química , Animales , Anticuerpos Monoclonales de Origen Murino/química , Toxinas Bacterianas/química , Línea Celular Tumoral , Exotoxinas/química , Femenino , Ratones , Ratones Endogámicos BALB C , Pseudomonas aeruginosa/química , Factores de Virulencia/química , Exotoxina A de Pseudomonas aeruginosaRESUMEN
AIM: Study the protective properties of "Staphylovac-2" vaccinie. MATERIALS AND METHODS: Samples of the vaccine manufactured by SPA "Microgen" based on the developed technology were studied in balb/c mice during 3- and 6-fold immunization schemes. Protective activity of the preparation was determined in experiments with active and passive protection during intraperitoneal infection, seeding of the causative agent from spleen and kidneys during intravenous infection, of animals. RESULTS: In experiments with active protection of mice for both 3- and 6-fold immunization schemes, a significant protective activity of the studied series was determined, compared with the control group of mice. Sera obtained after animal immunization (rabbits, mice) by staphylococcus vaccine had protective properties. A reduction of spleen and kidneys seeding by Staphylococcus aureus in immunized mice compared with the control group was detected in the model of generalized staphylococci infection. CONCLUSION: The preclinical studies carried out with the "Staphylovac-2" vaccine, developed baed on the complex of protective staplylococci antigens, have confirmed the high protective activity of the preparation.
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Antígenos Bacterianos/inmunología , Sueros Inmunes/administración & dosificación , Inmunización , Infecciones Estafilocócicas/prevención & control , Vacunas Estafilocócicas/inmunología , Staphylococcus aureus/inmunología , Inmunidad Adaptativa/efectos de los fármacos , Animales , Antígenos Bacterianos/administración & dosificación , Antígenos Bacterianos/química , Carga Bacteriana , Inmunidad Innata/efectos de los fármacos , Inyecciones Intraperitoneales , Inyecciones Subcutáneas , Riñón/efectos de los fármacos , Riñón/inmunología , Riñón/microbiología , Masculino , Ratones , Ratones Endogámicos BALB C , Conejos , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/microbiología , Infecciones Estafilocócicas/inmunología , Infecciones Estafilocócicas/microbiología , Vacunas Estafilocócicas/administración & dosificación , Vacunas Estafilocócicas/química , Staphylococcus aureus/químicaRESUMEN
Pneumonia often occurs as a secondary infection after influenza and accounts for a large proportion of the morbidity and mortality associated with seasonal and pandemic influenza outbreaks. The efficacy of umifenovir (Arbidol) was investigated on a murine model of S. aureus pneumonia following A/California/04/2009 (H1N1) influenza virusinfection. Oral treatment with umifenovir (40 and 60 mg/kg/day) in all the contamination schemes increased the survival rate in the mice from 0% to 90% and lowered the animal weight loss. The umifenovir treatment also decreased the virus titer by ≥ 2 logs and the viable bacteria counts in the lungs of the mice. The lungs of the mice treated with umifenovir had less severe histopathologic lesions compared to the control group.
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Antivirales/farmacología , Indoles/farmacología , Pulmón/efectos de los fármacos , Infecciones por Orthomyxoviridae/tratamiento farmacológico , Neumonía Bacteriana/tratamiento farmacológico , Infecciones Estafilocócicas/tratamiento farmacológico , Administración Oral , Animales , Carga Bacteriana/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Coinfección , Modelos Animales de Enfermedad , Esquema de Medicación , Femenino , Subtipo H1N1 del Virus de la Influenza A/efectos de los fármacos , Subtipo H1N1 del Virus de la Influenza A/crecimiento & desarrollo , Pulmón/microbiología , Pulmón/patología , Pulmón/virología , Ratones , Ratones Endogámicos BALB C , Infecciones por Orthomyxoviridae/mortalidad , Infecciones por Orthomyxoviridae/patología , Infecciones por Orthomyxoviridae/virología , Neumonía Bacteriana/microbiología , Neumonía Bacteriana/mortalidad , Neumonía Bacteriana/patología , Infecciones Estafilocócicas/microbiología , Infecciones Estafilocócicas/mortalidad , Infecciones Estafilocócicas/patología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Análisis de Supervivencia , Carga Viral/efectos de los fármacosRESUMEN
AIM: Study protective activity of S. pneumoniae protein-containing antigen complex obtained from T3No.3 strain against infection by homologous pneumococcus strain. MATERIALS AND METHODS: S. pneumoniae T3No.3 (serotype 3) strain obtained from collection of pneumococcus strains of Mechnikov Research Institute of Vaccines and Sera was used in the study. S. pneumoniae protein-containing antigen complex was isolated by precipitation by 2 volumes of acetone of supernatant fraction of cultural medium used for pneumococcus cultivation. Molecular mass of proteins contained in S. pneumoniae antigen complex was determined by SDS electrophoresis in polyacrylamide gel. Protective activity of S. pneumoniae protein-containing antigen complex was studied in BALB/c line mice active protection experiments. Activity of mice immune sera obtained against whole-cell pneumococcus culture (T3No.3 strain) was determined in vitro by solid phase indirect EIA. RESULTS: The data obtained give evidence that the isolated protein-containing antigen complex from S. pneumoniae T3No.3 strain effectively protects mice from consequent infection by a homologous S. pneumoniae strain. S. pneumoniae protein-containing antigen complex sorbed on solid phase at 5 microg dose was established by using EIA to interact with homologous mice immune sera. CONCLUSION: The results of the carried out studies allow to move to studies of cross-activity of S. pneumoniae protein-containing antigen complex isolated from T3No.3 strain.
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Antígenos Bacterianos/inmunología , Proteínas Bacterianas/inmunología , Infecciones Neumocócicas/inmunología , Vacunas Neumococicas/inmunología , Streptococcus pneumoniae/inmunología , Animales , Antígenos Bacterianos/química , Antígenos Bacterianos/aislamiento & purificación , Antígenos Bacterianos/farmacología , Proteínas Bacterianas/química , Proteínas Bacterianas/aislamiento & purificación , Proteínas Bacterianas/farmacología , Femenino , Masculino , Ratones , Ratones Endogámicos BALB C , Infecciones Neumocócicas/prevención & control , Vacunas Neumococicas/química , Vacunas Neumococicas/aislamiento & purificación , Vacunas Neumococicas/farmacologíaRESUMEN
AIM: Production of water soluble protein-containing antigens from various strains of S. pneumoniae during cultivation in complete and semi-synthetic culture media as well as selection of strains with cross antigenic activity. MATERIALS AND METHODS: S. pneumoniae 3, 6A, 6B, 14, 10A, 18A, 19A, 19F, 23F serotype strains were cultivated in brain-heart broth and semi-synthetic medium with addition of aminopeptide for 24 hours at 37 degrees C for the production of water soluble antigens. The antigens were obtained by a method of triple water extraction from acetone dried microbial cells. Chemical composition of preparations, electrophoresis mobility of protein-containing components of preparations and cross antigenic activity in gel immune diffusion reaction by using rabbit hyperimmune sera were studied. RESULTS: In studies of 10 pneumococcus strains from various serotypes a method of microbial cell inactivation by acetone was selected that allows to produce preparations with high protein content (25.5 - 53.1%). Electrophoretic separation of the preparations revealed difference in the preparations obtained from various pneumococcus strains in the layout of major protein lines in the 8 - 95 kDa range. The most virulent and immunogenic S. pneumoniae strain that during cultivation in semi-synthetic medium was characterized by intraspecies cross antigenic activity and in gel immune diffusion reacted with all the studied sera against 3, 14, 18C, 23F serotype strains was selected. CONCLUSION: The study resulted in the selection of a technologically simple method of production of pneumococcus antigens with high protein content and showed that only 1 of the studied preparations produced from a virulent strain with poorly expressed S. pneumoniae capsule during cultivation in semi-synthetic medium has the highest cross antigenic activity.