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The review presents the main physiological functions of thrombin. The procoagulant and anticoagulant activities of the key serine protease are discussed in both physiological and pathological conditions of hemostasis. The involvement of thrombin in atherogenesis, as well as its role as a mediator of vascular dysfunction and inflammation in both the peripheral and central nervous system, is highlighted. A pronounced imbalance between the pro- and anticoagulant systems leads to an increase in thrombin formation and creates conditions for the development of thrombosis. Tests that allow direct or indirect assessment of thrombin's functional activity are presented. The potential applications of direct thrombin inhibitors and direct blockers of thrombin PAR receptors in vascular neurology are also considered.
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Neurología , Trombina , Humanos , Serina Endopeptidasas , Anticoagulantes , Sistema Nervioso CentralRESUMEN
Toxoplasmosis is a zoonotic protozoal disease characterized by a chronic course, polymorphism of clinical manifestations, predominant damage to the central nervous system, organs of vision, liver and lungs. The causative agent of the disease is the obligate intracellular parasite Toxoplasma gondii, which circulates widely in the external environment and has a large circle of intermediate hosts. Toxoplasmosis is classified by the method of infection (congenital or acquired), by pathogenesis (acute or chronic), by manifestation (latent or with the manifestation of symptoms). According to the state of the human immune system, the disease can occur without immunodeficiency, while the patient has a chronic lifelong carrier, and with immunodeficiency. People with HIV most commonly present with cerebral toxoplasmosis. The article presents a case of the development of toxoplasmosis in a patient in the absence of a burdened history.
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Toxoplasma , Toxoplasmosis Cerebral , Humanos , Neurólogos , Toxoplasmosis Cerebral/diagnóstico , Sistema Nervioso Central , Polimorfismo GenéticoRESUMEN
Members of the genus Thermaerobacter belong to the phylum Firmicutes and all isolates characterised to date are strictly aerobic and thermophilic. They were isolated from a mud sample of the Challenger Deep in the Mariana Trench, hydrothermal vents, and silt compost. A novel thermophilic, facultatively lithoautotrophic bacteria of the genus Thermaerobacter, strain PB12/4term (=VKM B-3151T), with a metabolism that is uncharacteristic of the type species, was isolated from low-temperature surface sediments near the Posolsk Bank methane seep, Lake Baikal, Russia. The new strain grows with molecular hydrogen as electron donor, elemental sulfur, and thiosulfate as electron acceptors, and CO2/[Formula: see text] as carbon source. The genome of strain PB12/4term consists of one chromosome with a total length of 2.820.915 bp and the G+C content of the genomic DNA was 72.2%. The phylogenomic reconstruction based on 120 conserved bacterial single-copy proteins revealed that strain PB12/4term belongs to the genus Thermaerobacter within in the class Thermaerobacteria, phylum Firmicutes_E. The strain PB12/4term is closely related to Thermaerobacter subterraneus DSM 13965 (ANI=95.08%, AF=0.91) and Thermaerobacter marianensis DSM 12885 (ANI=84.98%, AF=0.77). Genomic and experimental data confirm the ability of the Thermaerobacter PB12/4term pure culture to facultatively lithotrophic growth, which is provided by the presence of [NiFe]hydrogenase enzymes that are absent in T. marianensis DSM 12885 and T. subterraneus DSM 13965. The data obtained on the physiological and biochemical differences of strain PB12/4term provide a deeper insight into the species diversity and functional activity of the genus Thermaerobacter.
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Bacterias Aerobias , Lagos , Temperatura , Lagos/análisis , Análisis de Secuencia de ADN , Bacterias Aerobias/genética , Genómica , Filogenia , ARN Ribosómico 16S/genética , ADN Bacteriano/genética , ADN Bacteriano/análisis , Técnicas de Tipificación BacterianaRESUMEN
In addition to exhibited antioxidant and anti-inflammatory activity, fullerene C60 is a promising wound healing agent. An important stage in the production of fullerene-based ointments is the stability of the aqueous fullerene dispersion (AFD) with minimum size of colloidal fullerene aggregates and sufficiently high concentration. To achieve these parameters tangential flow filtration of fullerene C60 was used ("green technology"). As estimated by small-angle neutron scattering and dynamic light scattering purified AFDs with narrow-size distribution nanoclusters have a size of 6 nm and are assembled into agglomerates which reach a size of 150 nm. The ability of the AFD to exhibit regenerative activity was studied using the animal wound model. This study shows for the first time that the fullerene-based composition stimulates the healing of wounds of various origins. We assume that the mechanism of the AFD wound-healing activity is associated with the aryl hydrocarbon receptor and macrophages activity.
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TecnologíaRESUMEN
In this study, we present the first results on oxidation stress in Lake Baikal phytoplankton and its adaptation to environmental changes under anthropogenic impact. As was shown, the changing of the dominant species of phytoplankton collected from the surface water layer (~0.3 m) took place from February to June 2021. Phytoplankton were collected at a nearshore station (a littoral station at a distance of ~0.01 km from the shoreline, depth to bottom is ~5 m) and an offshore station (a pelagic station at a distance of ~1 km from the shoreline, depth to bottom is ~543 m). In February, dinoflagellates were dominant (~40 %) as well as diatoms (≤33 %) and green algae (≤12 %). Their biomass was 100 mg·m-3. In March, chrysophytes were dominant (up to 50 %) as well as cryptophytes (≤43 %) and dinoflagellates (≤30 %). Their biomass was 160-270 mg·m-3. In April, biomass increased up to 700-3100 mg·m-3 with the dominance of large cell dinoflagellates (up to 99 %), chrysophytes (up to 50 %), and cryptophytes (up to 35 %). By the end of the first decade of May, the percentage of dinoflagellates decreased and that of cryptophytes increased. In the second decade of May, the percentage of diatoms increased up to ~26-38 % but phytoplankton biomass was minimal (13-30 mg·m-3). By June, the percentage of diatoms in the samples reached 44-75 % at 60-550 mg·m-3. The oxidation stress of phytoplankton as a nonspecific adaptive response to a prolonged, intensive, or recurrent effect of a stress factor was estimated from the content of thiobarbituric acid reactive substances (TBARS). The mean content of these substances (markers of the lipid peroxidation) was determined spectrophotometrically. The oxidation stress of phytoplankton was revealed only when diatom algae dominated. It can be explained by adaptation of algae of other classes to the stress factor. The content of the lipid peroxidation markers in the coastal phytoplankton collected close to the settlement of Listvyanka known as a large touristic center was estimated from 100 to 500 µg·g-1 of dry weight of sample. During the period of diatom blooming in 2016 and 2018, oxidation stress of phytoplankton collected near large settlements was found. In phytoplankton from deep-water pelagic stations most remote from settlements, stress was not revealed. Using the method of gas chromatography, we showed a lower (up to 15 %) content of polyunsaturated fatty acids in phytoplankton characterized by stress occurrence. This confirms cell membrane damages. In Lake Baikal surface water, we found a higher content of synthetic anionic surfactants (sodium alkylbenzene sulfonates), which are components of detergents and cause oxidation stress of hydrobionts (up to 30 ± 4 µg·L-1). The presence of these substances in a water ecosystem can result in exhausting of phytoplankton cell resources, homeostasis imbalance, stress, pathological changes, and rearrangements in phytoplankton assemblage.
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AIM OF THE STUDY: To investigate the efficacy and safety of non-immunogenic staphylokinase (NS) compared with alteplase (A) in patients with acute ischemic stroke (AIS) within 4.5 h after symptom onset. MATERIAL AND METHODS: 336 patients with IS within 4.5 h after symptom onset were included in a randomized, open-label, multicenter, parallel-group, non-inferiority comparative trial of NS vs A (168 patients in each group). NS was administered as an intravenous bolus in a dose of 10 mg, regardless of body weight, over 10 s, A was administered as a bolus infusion in a dose of 0.9 mg/kg, maximum 90 mg over 1 hour. The primary efficacy endpoint was a favorable outcome, defined as a modified Rankin scale (mRS) score of 0-1 on day 90. Safety endpoints included all-cause mortality on day 90, symptomatic intracranial haemorrhage, and other serious adverse events (SAEs). RESULTS: At day 90, 84 (50%) patients reached the primary endpoint (mRS 0-1) in the NS group, 68 (41%) patients - in the A group (p=0.10, OR=1.47, 95% CI=0.93-2.32). The difference between groups NS and A was 9.5% (95% CI= -1.7-20.7) and the lower limit of the 95% CI did not cross the margin of non-inferiority (pnon-inferiority<0.0001). There were no significant differences in the frequency of deaths between the groups: on day 90, 17 (10%) patients in the NS group and 24 (14%) in the A group had died (p=0.32). There was a trend towards significant differences in the frequency of symptomatic intracranial haemorrhage: NS group - 5 (3%) patients, A group - 13 (8%) patients (p=0.087, OR=0.37, 95% CI=0.1-1.13). There were significant differences in the number of patients with SAEs: in the NS group - 22 (13%) patients, in the A group - 37 (22%) patients (p=0.044, OR=0.53, 95% CI=0.28-0.98). CONCLUSION: The presented results of the FRIDA trial are the first in the world to use a drug based on NS in patients with IS. It has been shown that a single bolus (within 10 s) administration of NS at a standard dose of 10 mg, regardless of body weight, allows to conduct fast, effective and safe thrombolytic therapy in patients with IS within 4.5 h after symptom onset. In further clinical tials of NS, it is planned to expand the therapeutic window beyond 4.5 h after symptom onset in patients with IS.
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Isquemia Encefálica , Accidente Cerebrovascular Isquémico , Metaloendopeptidasas , Accidente Cerebrovascular , Peso Corporal , Isquemia Encefálica/complicaciones , Isquemia Encefálica/tratamiento farmacológico , Fibrinolíticos/uso terapéutico , Humanos , Hemorragias Intracraneales/inducido químicamente , Hemorragias Intracraneales/complicaciones , Metaloendopeptidasas/uso terapéutico , Accidente Cerebrovascular/tratamiento farmacológico , Accidente Cerebrovascular/etiología , Terapia Trombolítica , Resultado del TratamientoRESUMEN
The new coronavirus infection (COVID-19) represents a challenge for global health. Since the outbreak began, the number of confirmed cases has exceeded 117 million, with more than 2.6 million deaths worldwide. With public health measures aimed at containing the spread of the disease, several countries have faced a crisis in the availability of intensive care units. Currently, a large-scale effort is underway to identify the nucleotide sequences of the SARS-CoV-2 coronavirus that is an etiological agent of COVID-19. Global sequencing of thousands of viral genomes has revealed many common genetic variants, which enables the monitoring of the evolution of SARS-CoV-2 and the tracking of its spread over time. Understanding the current evolution of SARS-CoV-2 is necessary not only for a retrospective analysis of the new coronavirus infection spread, but also for the development of approaches to the therapy and prophylaxis of COVID-19. In this review, we have focused on the general characteristics of SARS-CoV-2 and COVID-19. Also, we have analyzed available publications on the genetic diversity of the virus and the relationship between the diversity and the biological properties of SARS-CoV-2, such as virulence and contagiousness.
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Gene therapy is a promising approach for personalized medicine, but its application in humans requires development of efficient and safe vehicles. PEGylated liposomes are some of the most suitable delivery systems for nucleic acids because of their stability under physiological conditions and prolonged circulation time, compared to conventional and other types of "stealth" liposomes. In vitro/in vivo activity of PEGylated liposomes is highly dependent on PEG motif abundance. The process of "stealth" coverage formation is a very important parameter for efficient transfection assays and further fate determination of the PEG layer after tissue penetration. In this review, we discuss the latest methods of PEGylated liposome preparation.
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BACKGROUND: Mouse models of allergic asthma play a crucial role in exploring of asthma pathogenesis and testing of novel anti-inflammatory drugs. Widely used acute asthma models usually developed with adjuvant (aluminum hydroxide (alum)) do not reproduce one of the main asthma feature - airway remodeling while chronic asthma model mimic the pathophysiology of human disease. Moreover, the use of alum causes distress in experimental animals and impedes the test of adjuvant-containing drugs. In this study, we aimed to develop a chronic adjuvant-free asthma model with pronounced asthmatic phenotype. METHODS: Female BALB/c mice were divided into 3 groups. The first group was sensitized with intraperitoneal injections of ovalbumin (OVA) emulsified in aluminum hydroxide on days 0, 14, 28 followed by two stages of intranasally challenge with OVA on days 41-43 and 62-64. The second group was subcutaneously sensitized with the same dose of OVA without adjuvant and challenged on the same days. The third group (negative control) included mice which did not received any kind of treatment (i.e. sensitization and challenge). Serum levels of OVA-specific IgE, IgG2a and IgG1 antibodies were detected by ELISA. Airway hyper-responsiveness was measured by non-invasive plethysmography on days 44 and 65. Bronchoalveolar lavage fluids (BALF) sampled in all groups on days 45 and 66 were analyzed by light microscopy. The left lung was removed for histological analysis. The IL-4 and IFNγ mRNA expression in BALF cells was evaluated by RT-PCR. RESULTS: The OVA-specific IgE antibody response was two-fold increased in mice from adjuvant-free group compared to the adjuvant group that reflects reorientation of immune response towards Th2 phenotype. At the same time, the level of OVA-specific IgG1 and IgG2a antibodies was increased in the adjuvant group. Airway hyperresponsiveness to methacholine in mice of both experimental groups was two-fold higher than in control. Analysis of cell composition in BAL has shown a significant increase in eosinophil count in both experimental groups that indicate the development of allergic inflammation. Lung histology revealed airway remodeling in both experimental groups including goblet cell hyperplasia/metaplasia, thickening of airway walls, collagen deposition in the wall of distal airways. Additionally, the tendency to develop hypertrophy of bronchial smooth muscle layer was observed. Study of gene expression in BAL cells revealed the increase of IL-4 level in both adjuvant and adjuvant-free groups while IFNγ expression in both experimental groups was similar to control group. CONCLUSION: We have developed a chronic adjuvant-free mouse asthma model which possesses all necessary features of the disease including airway remodeling and is more suitable for pre-clinical evaluation of novel therapeutic approaches including adjuvant-containing drugs.
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Adyuvantes Inmunológicos , Hidróxido de Aluminio , Asma/inducido químicamente , Pulmón , Ovalbúmina , Hipersensibilidad Respiratoria/inducido químicamente , Remodelación de las Vías Aéreas (Respiratorias) , Animales , Asma/sangre , Asma/inmunología , Asma/fisiopatología , Líquido del Lavado Bronquioalveolar/inmunología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Femenino , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Interferón gamma/genética , Interferón gamma/metabolismo , Interleucina-4/genética , Interleucina-4/metabolismo , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Pulmón/fisiopatología , Ratones Endogámicos BALB C , Hipersensibilidad Respiratoria/sangre , Hipersensibilidad Respiratoria/inmunología , Hipersensibilidad Respiratoria/fisiopatología , Células Th2/inmunología , Células Th2/metabolismo , Factores de TiempoRESUMEN
Interleukin-33 (IL-33) belongs to the IL-1 cytokine family and plays an important role in modulating immune system by inducing Th2 immune response via the ST2 membrane receptor. Epithelial cells are the major producers of IL-33. However, IL-33 is also secreted by other cells, e.g., bone marrow cells, dendritic cells, macrophages, and mast cells. IL-33 targets a broad range of cell types bearing the ST2 surface receptor. Many ST2-positive cells, such as Th2 cells, mast cells, basophils, and eosinophils, are involved in the development of allergic bronchial asthma (BA). This suggests that IL-33 directly participates in BA pathogenesis. Currently, the role of IL-33 in pathogenesis of inflammatory disorders, including BA, has been extensively investigated using clinical samples collected from patients, as well as asthma animal models. In particular, numerous studies on blocking IL-33 and its receptor by monoclonal antibodies in asthma mouse model have been performed over the last several years; IL-33- and ST2-deficient transgenic mice have also been generated. In this review, we summarized and analyzed the data on the role of IL-33 in BA pathogenesis and the prospects for creating new treatments for BA.
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Asma/inmunología , Asma/fisiopatología , Interleucina-33/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Asma/genética , Asma/patología , Humanos , Interleucina-33/antagonistas & inhibidores , Interleucina-33/deficiencia , Interleucina-33/genéticaRESUMEN
In the study, the effect of the TLR4 agonist Immunomax was investigated in vitro and in vivo. In particular, Immunomax was shown to polarize mouse bone marrow macrophages from the M0 and M2 states into the M1 state (ARG1 and iNOS mRNA expression levels were used to identify the mouse M1 and M2 phenotypes). Next, we investigated the prophylactic antiviral effect of Immunomax in both a model of mouse respiratory syncytial virus (RSV) infection and a model of RSV-induced bronchial asthma (BA) exacerbation. In the experiment with RSV-induced BA exacerbation, Immunomax-treated mice were characterized by a significant decrease of the viral load in lung homogenates, an increased amount of M1 macrophages in the lung, a tendency toward Th2-dependent ovalbumin-specific IgG1 antibodies decrease in blood serum, a significant increase in RSV-activated CD4+ T cells secreting IFNγ (Th1 cells), and a simultaneous significant decrease in the amount of CD4+ cells secreting IL-4 (Th2 cells) in the mouse spleen, which were detected by ELISPOT 1.5 months after experiment. These findings suggest that treatment with the TLR4 agonist Immunomax polarizes the immune response towards antiviral Th1 and may be used for short-term antiviral prophylaxis to prevent acute respiratory viral infections in asthmatics.
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The results of the study of the distribution of calicivirus infection in a population of domestic cats of different breeds, contained individually or the group method, the virus isolation in the cell culture and a comparative phylogenetic analysis of their nucleotide sequences with published sequences of reference feld and vaccine strains of Feline calicivirus (FCV) from other countries: USA, Germany, Japan, China and Korea are presented. Clinical signs of infection were found in 14.3% of the animals examined. After several passages in the primary kidney cells of the kitten embryo, seven cytopathogenic isolates FCV were isolated: 1 - from a cat with an acute infection, 5 - subclinical infection, 1 - systemic infection. They were adapted to continuous FK-81 cells in which they reached a maximum infectious activity of 10.0 ± 1.15 lg TCD 50 / cm3. Based on the sequence analysis of the open reading frame 2 region of the viral genome Eshli strain showed a close relationship with strain KM016908 from China with the identity of the nucleotide sequences between them of 81.0%. The results of the investigations showed that FCV isolates obtained from animals on the territory of Siberia are genetically different from strains included to imported vaccines used to prevent disease in Russian Federation and also among themselves. This causes a decrease in the effectiveness of preventive measures. In nurseries that do not have contacts and connections between themselves but located in the same geographic region FCV populations may have some genetic differences. A close relationship of some feld isolates with strains from other countries geographically located so far from the Siberian region has been revealed. Studies on the molecular epizootology of caliciviruses are important in the development of test systems and the monitoring of the spread of strains in Russia.
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Infecciones por Caliciviridae/genética , Calicivirus Felino/genética , Filogenia , Animales , Secuencia de Bases/genética , Infecciones por Caliciviridae/virología , Calicivirus Felino/aislamiento & purificación , Calicivirus Felino/patogenicidad , Gatos , República de Corea , Federación de Rusia/epidemiología , Siberia/epidemiología , Vacunas Virales/inmunología , Vacunas Virales/uso terapéuticoRESUMEN
Asthma is responsible for approximately 25,000 deaths annually in Europe despite available medicines that maintain asthma control and reduce asthma exacerbations. Better treatments are urgently needed for the control of chronic asthma and reduction in asthma exacerbations, the major cause of asthma mortality. Much research spanning >20 years shows a strong association between microorganisms including pathogens in asthma onset, severity and exacerbation, yet with the exception of antibiotics, few treatments are available that specifically target the offending pathogens. Recent insights into the microbiome suggest that modulating commensal organisms within the gut or lung may also be a possible way to treat/prevent asthma. The European Academy of Allergy & Clinical Immunology Task Force on Anti-infectives in Asthma was initiated to investigate the potential of anti-infectives and immunomodulators in asthma. This review provides a concise summary of the current literature and aimed to identify and address key questions that concern the use of anti-infectives and both microbe- and host-based immunomodulators and their feasibility for use in asthma.
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Antiasmáticos/uso terapéutico , Antiinfecciosos/uso terapéutico , Asma/tratamiento farmacológico , Asma/patología , Factores Inmunológicos/uso terapéutico , Factores de Edad , Antiasmáticos/administración & dosificación , Antiinfecciosos/administración & dosificación , Asma/etiología , Progresión de la Enfermedad , Femenino , Humanos , Factores Inmunológicos/administración & dosificación , Inmunomodulación/efectos de los fármacos , Masculino , Embarazo , Complicaciones del Embarazo , Probióticos/administración & dosificación , Resultado del Tratamiento , Vacunas/administración & dosificación , Vacunas/inmunologíaRESUMEN
To address uncertainties in the prevention and management of influenza in people with asthma, we performed a scoping review of the published literature on influenza burden; current vaccine recommendations; vaccination coverage; immunogenicity, efficacy, effectiveness, and safety of influenza vaccines; and the benefits of antiviral drugs in people with asthma. We found significant variation in the reported rates of influenza detection in individuals with acute asthma exacerbations making it unclear to what degree influenza causes exacerbations of underlying asthma. The strongest evidence of an association was seen in studies of children. Countries in the European Union currently recommend influenza vaccination of adults with asthma; however, coverage varied between regions. Coverage was lower among children with asthma. Limited data suggest that good seroprotection and seroconversion can be achieved in both children and adults with asthma and that vaccination confers a degree of protection against influenza illness and asthma-related morbidity to children with asthma. There were insufficient data to determine efficacy in adults. Overall, influenza vaccines appeared to be safe for people with asthma. We identify knowledge gaps and make recommendations on future research needs in relation to influenza in patients with asthma.
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Asma/complicaciones , Asma/epidemiología , Gripe Humana/complicaciones , Gripe Humana/epidemiología , Gripe Humana/prevención & control , Antivirales/uso terapéutico , Costo de Enfermedad , Salud Global , Humanos , Inmunogenicidad Vacunal , Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/efectos adversos , Vacunas contra la Influenza/inmunología , Gripe Humana/terapia , Evaluación del Resultado de la Atención al Paciente , Vigilancia en Salud Pública , Resultado del Tratamiento , VacunaciónRESUMEN
Bovine viral diarrhea virus (BVDV) belongs to the genus Pestivirus, family Flaviviridae. It causes various clinical forms of infection leading to significant economic losses in beef and dairy industry worldwide. Furthermore, the virus is a contaminant of biological preparations (bovine fetal serum, continuous cell cultures, vaccines for human and veterinary medicine, interferons, trypsin, biotechnological preparations, embryos, stem cells, etc.). It is used as a test object when developing methods of decontamination. In some countries, a tool for monitoring the infection caused by the virus is vaccination based on the use of live and inactivated vaccines with varying efficiency. The antiviral compounds are a potential means of control in case of insufficient efficacy of vaccines. Their advantage for BVDV control is the ability to provide immediate protection for animals at risk in the case of an outbreak of the disease. This review summarizes the current state of knowledge about antiviral compounds against BVDV. It was noted that due to the use of advanced biomedical technologies there is a tendency to search for drugs that might be effective for antiviral therapy of BVDV, as indicated by numerous studies of new compounds and the antiviral efficacy of known drugs used in medical practice. In addition to the well-known antiviral targets for the virus, such as the RdRp, IMPDH, NS3, new targets were discovered, such as protein p7. Its mechanism of action remains to be explored. It can be concluded that there is a great potential for BVDV control through the use of antiviral drugs which has not yet implemented. The biggest obstacle for commercial implementation of identified compounds is the lack of demonstration of their efficacy in vivo. Further studies should be performed to develop a method for administering effective drugs to groups of animals.
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Nucleic acid-based therapeutics have recently emerged as a new class of next generation agents for treatment and prevention of viral infection, cancer, and genetic disorders, but their wide use is limited by their relatively weak delivery into target cells. Usage of synthetic cationic amphiphiles with peptide hydrophilic domain as agents for non-viral gene delivery is an attractive approach. We developed the schemes for the synthesis of aliphatic peptides with different length of the hydrocarbon chains in hydrophobic domains and different amino acids in polar head. For the obtained derivatives we determined transfection efficiency, critical vesicle concentration, particle size, ζ-potential and aggregates stability. We have found that the transfection efficiency is increased if the ornithine is a part of polar head in an amphiphile. The most promising amphiphile for liposomal formation OrnOrnGlu(C16H33)2 was examined more carefully. It has been shown that the lipopeptide possesses low toxicity (in vitro and in vivo) and high transfection efficiency with pDNA and siRNA in different cell lines. In addition, the production of liposomes based on this lipopeptide is simple, quick and cheap. Thus OrnOrnGlu(C16H33)2 is a promising vehicle for gene delivery and gene silencing.
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Silenciador del Gen/efectos de los fármacos , Lipopéptidos/administración & dosificación , Lipopéptidos/química , Células A549 , Animales , Células CHO , Cationes/administración & dosificación , Cationes/química , Línea Celular , Línea Celular Tumoral , Cricetulus , ADN/metabolismo , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Células HEK293 , Células HeLa , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Células Jurkat , Liposomas/administración & dosificación , Liposomas/química , Tamaño de la Partícula , Plásmidos/administración & dosificación , Plásmidos/química , ARN Interferente Pequeño/metabolismo , Transfección/métodosRESUMEN
In this study, a rapid quantitative method using TaqMan-based real-time reverse transcription-polymerase chain reaction (qPCR-RT) has been developed for estimating the titers of measles, mumps and rubella (MMR) viruses in infected cell culture supernatants. The qPCR-RT assay was demonstrated to be a specific, sensitive, efficient and reproducible method. For MMR viral samples obtained during MMR viral propagations in Vero cells at a different multiplicity of infection, titers determined by the qPCR-RT assay have been compared with estimates of infectious virus obtained by a traditional commonly used method for MMR viruses - 50% cell culture infective dose (CCID(50)) assay, in paired samples. Pearson analysis evidenced a significant correlation between both methods for a certain period after viral inoculation. Furthermore, the established qPCR-RT assay was faster and less-laborious. The developed method could be used as an alternative method or a supplementary tool for the routine titer estimation during MMR vaccine production.
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Virus del Sarampión/aislamiento & purificación , Virus de la Parotiditis/aislamiento & purificación , ARN Viral/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Virus de la Rubéola/aislamiento & purificación , Animales , Línea Celular , Chlorocebus aethiops , Humanos , Sarampión/diagnóstico , Sarampión/virología , Virus del Sarampión/genética , Paperas/diagnóstico , Paperas/virología , Virus de la Parotiditis/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Rubéola (Sarampión Alemán)/diagnóstico , Rubéola (Sarampión Alemán)/virología , Virus de la Rubéola/genética , Sensibilidad y Especificidad , Células Vero , Carga ViralRESUMEN
The paper gives the results of a comparative analysis of the detection of influenza viruses in clinical samples, by using multiplex real-time polymerase chain reaction (RT-PCR) and by virus isolation in MDCK cell cultures. The investigation employed 267 nasopharyngeal swab specimens obtained from patients with influenza symptoms during two epidemic seasons (2008-2009 and 2009-2010). Influenza viruses were found in 104 samples (48 with influenza A virus (IAV) and 56 with influenza B virus (IBV)) by multiplex RT-RCR and in 84 samples (35 with IAV and 49 with IBV) by a cultural technique. The results of detection of influenza viruses by the two methods showed 89.4% agreement. The diagnostic sensitivity of multiplex RT-PCR testing a panel of the clinical samples in question was estimated to be 94.3% for IAV and 95.9% for IBV. The diagnostic sensitivity of multiplex RT-PCR in virus detection was demonstrated to be not only highly competitive with virus isolation, but also superior to the latter.
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Virus de la Influenza A/genética , Virus de la Influenza B/genética , Gripe Humana/diagnóstico , Animales , Técnicas de Cultivo de Célula , Línea Celular , Diagnóstico Diferencial , Perros , Femenino , Humanos , Virus de la Influenza A/aislamiento & purificación , Virus de la Influenza B/aislamiento & purificación , Gripe Humana/virología , Masculino , Nasofaringe/virología , Reacción en Cadena en Tiempo Real de la Polimerasa , Sensibilidad y EspecificidadRESUMEN
Carriage frequencies of alleles and genotypes of polymorphous locus of -174G>C IL6 (rs1800795) were analyzed in the patients with ischemic stroke (IS) of Russian ethnic descent (200 cases) and in the control group of the same ethnic descent with similar sex and age (140 controls). Significant differences were identified in frequencies of carriage (in homo- or heterozygous form) of allele IL6*-174G (p = 0.0029, OR = 2.9, 95% CI: 1.4-5.8), which can be considered as risk factor for IS and in frequencies of IL6*-174C/C genotype carriage, correspondingly (p = 0.0029, OR = 0.35, 95% CI: 0.17-0.69). After sex stratification of patients and controls similar significant differences were observed only between female patients and controls, after age stratification the difference was observed only for the age group older 60 years. Complex analysis of association of SNP -174G>C IL6 alleles and genotypes carriage in combination with SNP 4266A>G (Thr312Ala) FGA (rs6050) (see symbol) -249C>T FGB (rs1800788) with IS revealed protective combinations IL6*-174C/C + FGA* 4266A (see symbol) IL6*-174C/C + FGB*-249C, which were slightly more significant than single protective genotype IL6*-174C/C associated with IS and their ORs didn't differ substantially from the single genotypes's OR value. At the same time the combinations of alternative allele IL6*-174G with the same FGB*-249C or FGA* 4266A alleles were revealed and their association significance levels as well as OR values were lower than the values for the single risk allele IL6*-174G. In case of the mutual carriage of IL6*-174G allele with FGA*4266A/A, FGB*-249C/C genotypes or with combinations of these alleles/genotypes the "neutralized" effect became stronger. In other words, we observed association of IS with allele/genotype combinations of genes IL6, FGA and FGB, in which IL6 plays key role and FGA and FGB have modulating function. In analysis of association of fibrinogen plasma levels with three analyzed polymorphous loci significant differences were not revealed.
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Fibrinógeno/genética , Fibrinógeno/metabolismo , Interleucina-6/genética , Polimorfismo de Nucleótido Simple/genética , Accidente Cerebrovascular/genética , Anciano , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Factores de RiesgoRESUMEN
The cytokine interleukin (IL)-15, major histocompatibility complex (MHC) class I molecules and MHC class I chain-related proteins (MIC) A and B are involved in cellular immune responses to virus infections but their role in respiratory syncytial virus (RSV) infection has not been studied. We aimed to determine how RSV infection modulates IL-15 production, MHC class I and MICA expression in respiratory epithelial cells, the molecular pathways implicated in virus-induced IL-15 production and how interferon (IFN)-γ alters RSV-induced IL-15 production and MHC class I and MICA expression. We infected respiratory epithelial cell lines (A549 and BEAS-2B cells) and primary bronchial epithelial cells with RSV and measured production of IL-15, expression of MHC I and MICA and the role of the transcription factor nuclear factor (NF)-κB. We report here that RSV increases IL-15 in respiratory epithelial cells via virus replication and NF-κB-dependent mechanisms. Furthermore, RSV infection of epithelial cells upregulated cell surface expression of MICA and levels of soluble MICA. IFN-γ upregulated RSV induction of soluble IL-15 but inhibited induction of MICA. Upregulation of IL-15, MHC I and MICA are likely to be important mechanisms in activating immune responses to RSV by epithelial cells.