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Orf or contagious ecthyma is a highly contagious, zoonotic, and economically important global viral disease of small ruminants and is endemic in India. Vaccination of susceptible goats/sheep along with suitable recombinant protein-based serological assay will be useful in the control of the infection. In this study, the full-length and truncated versions of F1L encoding gene (ORF 059) of orf virus were cloned into pFasBac HT A vector, transformed in DH10Bac cells, and expressed in insect cells. The full-length and truncated recombinant F1L proteins were expressed as a 6 × histidine-tagged fusion protein for ease of purification by Ni-NTA affinity chromatography under denaturing conditions. A protein with ~ 40 kDa and ~ 35 kDa for full-length and truncated F1L protein, respectively, were expressed and confirmed by SDS-PAGE and western blot. The protein reactivity evaluated by western blot analysis and indirect ELISA using ORFV hyperimmune serum was also found to be reactive. The results of the present study showed that the purified recombinant F1L protein can be used as a diagnostic antigen in sero-surveillance of ORFV infection in small ruminants. To the best of authors' knowledge, this is the first report on the expression of ORFV F1L in insect cells using a baculovirus vector and its successful purification to use as the potential diagnostic antigen in ELISA.
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Radial artery occlusion (RAO) is now a major concern in transradial approach (TRA). RAO limits future radial artery use for further TRA, for as a conduit during CABG, for invasive hemodynamic monitoring and for creation of arteriovenous fistula for hemodialysis in Chronic Kidney Disease (CKD) patients through same vascular approach. The effect of duration of hemostatic compression of RAO is unknown in Bangladesh. This prospective observational study was conducted in the department of Cardiology, National Institute of Cardiovascular Diseases, Dhaka, Bangladesh (NICVD) from September 2018 to August 2019, to evaluate the effect of duration of hemostatic compression on the incidence of radial artery occlusion (RAO) after transradial percutaneous coronary intervention. A total of 140 patients underwent percutaneous coronary intervention (PCI) through TRA. RAO was defined as an absence of antegrade flow or monophasic flow or invert flow on Duplex study. In this study 70 patients (Group I) received 2 hours hemostatic compression after transradial PCI. Another 70 patients (Group II) received 6 hours hemostatic compression after transradial PCI. Radial arterial blood flow assessed at early (24 hours) and late (30 days) by color duplex study after the procedure in both groups. Early radial artery occlusion occurred in 4.3% of patients in Group I and 12.8% of patients in Group II, (p=0.04). Late radial artery occlusion occurred in 2.8% of patients in Group I and 11.4% of patients in Group II, the difference was statistically significant (p=0.04). From multivariate logistic regression analysis duration of hemostatic compression time 6 hours (p=0.01), post-procedural nitroglycerine use (p=0.03) and procedure time (p=0.03) were predictors of RAO. Shorter duration of hemostatic compression is associated with a lower incidence of early and late radial artery occlusion after transradial intervention.
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Arteriopatías Oclusivas , Hemostáticos , Intervención Coronaria Percutánea , Humanos , Intervención Coronaria Percutánea/efectos adversos , Intervención Coronaria Percutánea/métodos , Arteria Radial/cirugía , Bangladesh , Arteriopatías Oclusivas/etiología , Arteriopatías Oclusivas/epidemiología , Cateterismo Cardíaco/métodos , Angiografía Coronaria/métodos , Resultado del TratamientoRESUMEN
BACKGROUND: Modern day otolaryngology has expanded beyond the ear, nose and throat to include head and neck surgery and aesthetic facial procedures. Photographic documentation is important within this expanded horizon. The spectrum of clinical photography includes photomicrographs, endoscopic photographs, peri-operative photography and medical social photography. METHOD: This article aimed to review the standard guidelines essential to obtain, store and disseminate photographs and looked at setting up a small clinic with minimal gadgets to suit clinical photography requirements. Elaboration of basic photography techniques in otolaryngology was reviewed, with examples of photographs taken in a clinic by a clinician. Advances and innovation in clinical photography, in the form of smartphone photography, artificial intelligence, device editing and newer hardware and software in otorhinolaryngology was reviewed. CONCLUSION: Having a professional photographer to aid a clinician is a luxury. Simple knowledge and regular practice of basic photography guidelines by a clinician is imperative.
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Inteligencia Artificial , Otorrinolaringólogos , Humanos , Fotograbar , Teléfono Inteligente , Documentación/métodosRESUMEN
The vapor pressure values of common elements are available in the literature over a limited temperature range and the accuracy and reliability of the reported data are not generally available. We evaluate the reliability and uncertainty of the available vapor pressure versus temperature data of fifty common pure elements and recommend vapor pressure versus temperature relations. By synthesizing the vapor pressure values from measurements reported in the literature with the values computed using the Clausius Clapeyron relation beyond the boiling point, we extend the vapor pressure range from 10-8 atm to 10 atm. We use a genetic algorithm to optimize the fitting of the vapor pressure data as a function of temperature over the extended vapor pressure range for each element. The recommended vapor pressure values are compared with the corresponding literature values to examine the reliability of the recommended values.
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Goatpox is an economically significant transboundary viral disease of goats that is caused by goatpox virus (GTPV). This study describes the prokaryotic expression of the GTPV ORF117 protein, a homologue of vaccinia virus A27L, and evaluation of its diagnostic potential in ELISA. The GTPV ORF117 gene was cloned into the pET32a vector to express recombinant ORF117 protein (rA27L) in E. coli BL21-CodonPlus (DE3)-RIPL. The bacterial expression of the protein was confirmed by western blot analysis using anti-GTPV polyclonal antibodies that detected rA27L, which is ~ 35 kDa in size. rA27L was affinity purified under native conditions and used to assess the antibody response in an optimized indirect ELISA. The purified antigen specifically reacted with anti-GTPV and anti-SPPV serum in ELISA. A preliminary screening of random and purposive serum samples (n = 520) from sheep and goats using this optimized ELISA gave a positivity rate of 19.4 % with a diagnostic specificity of 88.7% and diagnostic sensitivity of 98.5% when compared to the gold standard serum neutralization test. Our results suggest that the indirect ELISA based on the rA27L protein has potential for serosurveillance and seromonitoring of GTPV in goats.
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Antígenos Virales/análisis , Antígenos Virales/genética , Capripoxvirus/metabolismo , Ensayo de Inmunoadsorción Enzimática/métodos , Enfermedades de las Cabras/virología , Infecciones por Poxviridae/veterinaria , Proteínas Virales/análisis , Proteínas Virales/genética , Animales , Antígenos Virales/aislamiento & purificación , Antígenos Virales/metabolismo , Capripoxvirus/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Enfermedades de las Cabras/diagnóstico , Cabras , Infecciones por Poxviridae/diagnóstico , Infecciones por Poxviridae/virología , Proteínas Virales/aislamiento & purificación , Proteínas Virales/metabolismoRESUMEN
Land use and land cover (LULC) change have considerable influence on ecosystem services. Assessing change in ecosystem services due to LULC change at different spatial and temporal scales will help to identify suitable management practices for sustaining ecosystem productivity and maintaining the ecological balance. The objective of this study was to investigate variations in ecosystem services in response to LULC change over 27 years in four agro-climatic zones (ACZ) of eastern India using satellite imagery for the year 1989, 1996, 2005, 2011 (Landsat TM) and 2016 (Landsat 8 OLI). The satellite images were classified into six LULC classes, agriculture land, forest, waterbody, wasteland, built-up, and mining area. During the study period (1989 to 2016), forest cover reduced by 5.2%, 13.7%, and 3.6% in Sambalpur, Keonjhar, and Kandhamal districts of Odisha, respectively. In Balasore, agricultural land reduced by 17.2% due to its conversion to built-up land. The value of ecosystem services per unit area followed the order of waterbodies > agricultural land > forests. A different set of indicators, e.g., by explicitly including diversity, could change the rank between these land uses, so the temporal trends within a land use are more important than the absolute values. Total ecosystem services increased by US$ 1296.4 × 105 (50.74%), US$ 1100.7 × 105 (98.52%), US$ 1867 × 105 (61.64%), and US$ 1242.6 × 105 (46.13%) for Sambalpur, Balasore, Kandhamal, and Keonjhar, respectively.
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Cambio Climático , Conservación de los Recursos Naturales/métodos , Monitoreo del Ambiente/métodos , Agricultura/métodos , Ecosistema , Bosques , India , Minería , Imágenes Satelitales/métodosRESUMEN
Catechol, one of the major effluents released by various chemical and metal processing industries, causes severe pollution of groundwater. Monitoring of catechol in water using cost-effective, handheld sensor is demanding for the safety of the environment. In this work, non-enzymatic zinc oxide thin film based electrochemical strip sensor is developed on conducting glass substrate for detection of catechol. The preparation of strip without employing standard Pt or Ag/AgCl electrodes and simply depositing ZnO through wet chemical process represents a cost-effective innovative technique. The ZnO thin film is characterized using field emission scanning electron microscope (FESEM), energy dispersive X-ray spectroscopy (EDS), atomic force microscopy (AFM) and grazing incidence X-ray diffractometer (GIXRD). Catechol is electrochemically detected by means of cyclic voltammetry and amperometry. A prominent redox peak of the developed strip attributed to the detection of catechol is observed at -0.26â¯V in cyclic voltammetry. The strip is integrated with readout meter and an algorithm is built based on the experimentally observed linear variation of amperometric current with catechol concentration. The quantitative detection performance is demonstrated by testing 0.1-12â¯ppm catechol solutions.
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Técnicas Biosensibles , Catecoles/aislamiento & purificación , Agua/química , Óxido de Zinc/química , Catecoles/química , Técnicas Electroquímicas , Microscopía de Fuerza Atómica , Oxidación-ReducciónRESUMEN
Background Simple and mucopurulent chronic bronchitis (SMCB) is characterized by recurrent mucoid or mucopurulent expectoration in absence of localized suppurative disease. This observational open label study was undertaken to evaluate the effects of homeopathic medicine in SMCB. Methods 1902 patients were screened from 07 centres out of which 1305 were excluded. 597 patients were enrolled as per the inclusion and exclusion criteria. A total of 14 pre-defined homeopathic medicines were shortlisted for prescription after repertorizing the pathological symptoms of SMCB. Outcomes were assessed through chronic bronchitis symptom scale (CBSS) and FEV1/ FVC ratio with spirometry for over a period of two years. Appearance of any change (relief/ worse)/ status quo was immediately followed by placebo/ change in dilution/ change in remedy. Statistical analysis was done using SPSS version 20. Results: 532 patients were analyzed based on the intention to treat principle using last observation carry forward method. Mean CBS score reduced from 29.86±4.5 at baseline to 12.33±7.6 at completion of 2 years. Repeated measures ANOVA, at time points 0 (baseline), 3, 6, 9, 12 ,15, 18, 21 and 24 months, showed significant reduction in CBS scores [Wilk's Lambda 0.104, F=564, df 524; p=00001]. The FEV1 and FEV1/FVC was maintained within normal limits. 86% prescriptions included Lycopodium, Arsenicum album, Pulsatilla, Phosphorus, Stannum metallicum, Calcarea carbonica, Silicea, Bryonia alba. Conclusion: The result suggests effectiveness of homeopathic treatment in early years of SCMB patients. Controlled trials are warranted. (AU)
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Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Bronquitis Crónica/terapia , Homeopatía , Pulsatilla nigricans/uso terapéutico , Arsenicum Album/uso terapéutico , Calcarea Carbonica/uso terapéutico , Lycopodium clavatum/uso terapéutico , Silicea Terra/uso terapéutico , Stannum Metallicum/uso terapéutico , BryoniaRESUMEN
An integrative functional genomics study of multiple forms of data are vital for discovering molecular drivers of cancer development and progression. Here, we present an integrated genomic strategy utilizing DNA methylation and transcriptome profile data to discover epigenetically regulated genes implicated in cancer development and invasive progression. More specifically, this analysis identified fibromodulin (FMOD) as a glioblastoma (GBM) upregulated gene because of the loss of promoter methylation. Secreted FMOD promotes glioma cell migration through its ability to induce filamentous actin stress fiber formation. Treatment with cytochalasin D, an actin polymerization inhibitor, significantly reduced the FMOD-induced glioma cell migration. Small interfering RNA and small molecule inhibitor-based studies identified that FMOD-induced glioma cell migration is dependent on integrin-FAK-Src-Rho-ROCK signaling pathway. FMOD lacking C-terminus LRR11 domain (ΔFMOD), which does not bind collagen type I, failed to induce integrin and promote glioma cell migration. Further, FMOD-induced integrin activation and migration was abrogated by a 9-mer wild-type peptide from the FMOD C-terminus. However, the same peptide with mutation in two residues essential for FMOD interaction with collagen type I failed to compete with FMOD, thus signifying the importance of collagen type I-FMOD interaction in integrin activation. Chromatin immunoprecipitation-PCR experiments revealed that transforming growth factor beta-1 (TGF-ß1) regulates FMOD expression through epigenetic remodeling of FMOD promoter that involved demethylation and gain of active histone marks with a simultaneous loss of DNMT3A and EZH2 occupancy, but enrichment of Sma- and Mad-related protein-2 (SMAD2) and CBP. FMOD silencing inhibited the TGF-ß1-mediated glioma cell migration significantly. In univariate and multivariate Cox regression analysis, both FMOD promoter methylation and transcript levels predicted prognosis in GBM. Thus, this study identified several epigenetically regulated alterations responsible for cancer development and progression. Specifically, we found that secreted FMOD as an important regulator of glioma cell migration downstream of TGF-ß1 pathway and forms a potential basis for therapeutic intervention in GBM.
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Epigénesis Genética , Epigenómica , Fibromodulina/genética , Regulación Neoplásica de la Expresión Génica , Genes Esenciales , Glioma/genética , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Ensamble y Desensamble de Cromatina , Colágeno Tipo I/metabolismo , Citoesqueleto/metabolismo , Metilación de ADN , Epigenómica/métodos , Proteína-Tirosina Quinasas de Adhesión Focal/metabolismo , Perfilación de la Expresión Génica , Estudio de Asociación del Genoma Completo , Glioma/metabolismo , Glioma/mortalidad , Humanos , Estimación de Kaplan-Meier , Modelos Biológicos , Pronóstico , Regiones Promotoras Genéticas , Transducción de Señal/efectos de los fármacos , Transcripción Genética , Transcriptoma , Factor de Crecimiento Transformador beta1/metabolismo , Quinasas Asociadas a rho/metabolismo , Familia-src Quinasas/metabolismoRESUMEN
Eclampsia is an important cause of maternal mortality in developing countries. This cross-sectional comparative study was conducted in the Departments of Clinical Pathology, Bangabandhu Sheikh Mujib Medical University (BSMMU), and Obstetrics and Gynecology, BSMMU, and Dhaka Medical College Hospital, Dhaka, Bangladesh, from March 2012 to February 2013 and was designed to evaluate liver markers to predict preeclampsia. One hundred fifty (150) women with pregnancy of ≥28 weeks, 50 for each normal, preeclampsia and eclampsia group, were enrolled purposively. The protocol was approved by IRB of BSMMU. Consent was taken from each patient. Serum total Bilirubin and ALT were assessed. Data was collected in a questionnaire and analyzed by SPSS-16. Quantitative data were compared by ANOVA or 't' test and qualitative data by chi-square test. P value <0.05 was considered significant. The patients of all groups were similar in age and gravida. The mean±SD serum total bilirubin and ALT were significantly higher in preeclampsia and eclampsia groups than normal pregnancy. However they were similar in preeclampsia and eclampsia group.
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Alanina Transaminasa/sangre , Bilirrubina/sangre , Eclampsia/sangre , Preeclampsia/sangre , Adulto , Bangladesh , Biomarcadores/sangre , Estudios Transversales , Femenino , Humanos , Pruebas de Función Hepática , Embarazo , Adulto JovenRESUMEN
We have prepared biocompatible composites of 80wt% polyvinyl alcohol (PVA)-(20wt%) polyvinylpyrrolidone (PVP) blend with different concentrations of bioactive nanohydroxyapatite, Ca10(PO4)6(HO)2 (HAP). The composite films demonstrated maximum effective conductivity (σâ¼1.64×10(-4)S/m) and effective dielectric constant (εâ¼290) at percolation threshold concentration (â¼10wt% HAP) at room temperature. These values of σ and ε are much higher than those of PVA, PVP or HAP. Our preliminary observation indicated excellent biocompatibility of the electrospun fibrous meshes of two of these composites with different HAP contents (8.5 and 5wt% within percolation threshold concentration) using NIH 3T3 fibroblast cell line. Cells viability on the well characterized composite fibrous scaffolds was determined by MTT [3-(4,5-di-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay analysis. Enhancement of σ, due to HAP addition, was found to show increased biocompatibility of the fibrous scaffold. Enhanced σ value of the PVA/PVP-HAP composite provided supporting cues for the increased cell viability and biocompatibility of the composite fibrous meshes. Excellent biocompatibility these electrospun composite scaffolds made them to plausible potential candidates for tissue engineering or other biomedical applications.
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Materiales Biocompatibles/química , Durapatita/química , Nanocompuestos/química , Alcohol Polivinílico/química , Povidona/química , Andamios del Tejido , Animales , Materiales Biocompatibles/farmacología , Supervivencia Celular/efectos de los fármacos , Durapatita/farmacología , Conductividad Eléctrica , Técnicas Electroquímicas , Ratones , Células 3T3 NIH , Alcohol Polivinílico/farmacología , Porosidad , Povidona/farmacología , Ingeniería de TejidosRESUMEN
The isolation of BTV-1 serotype from cattle in India and its phylogenetic relationship based on VP2 gene has been reported. Virus (JBP42/12/Ind) is isolated in BHK-21 cell line from blood sample tested positive for BTV antigen in sandwich ELISA from asymptomatic cattle. Full length VP2 gene of cattle isolate was amplified, cloned and sequenced. On BLAST analysis virus isolate was identified as BTV-1 serotype. Phylogenetic tree based on complete VP2 coding region segregated Indian isolates, Australian isolates and African/European isolates in three distinct clusters. Segregation of Indian BTV-1 isolates at close proximity to the monophyletic cluster of Australian BTV-1 isolates indicates the present isolate as "eastern topotype' of BTV. Multiple alignments of VP2 gene nucleotide sequences suggest that, Indian BTV-1 isolate is more closely related to Australian BTV-1 isolates; where 14.1% to 14.4% and 6.8% to 7.4% divergence was observed at nucleotide and deduced amino acid sequence level respectively.
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Herein, chlorophyll and betalain dyes are extracted from fresh spinach leaves and beetroots. Fourier transform infrared spectra are used to identify the characteristic peaks of the extracted dyes. UV-vis light absorption characteristics of the dyes and their mixed counterpart are investigated by varying their pH and temperature. These dyes are used as photo sensitizer for fabrication of zinc oxide photo-anode based dye sensitized solar cells (DSSCs). The photo-voltaic characteristics of the developed DSSCs are measured under simulated solar light (power of incident light 100 mW cm(-2) from Air Mass 1.5G). The solar to electric conversion efficiencies for the chlorophyll, betalain and mixed dye based solar cells are estimated as 0.148%, 0.197% and 0.294% respectively. The highest conversion efficiency for mixed dye based solar cell is attributed due to the absorption of wider range of solar spectrum.
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Beta vulgaris/química , Betalaínas/química , Clorofila/química , Suministros de Energía Eléctrica , Fármacos Fotosensibilizantes/química , Spinacia oleracea/química , Betalaínas/aislamiento & purificación , Clorofila/aislamiento & purificación , Colorantes/química , Colorantes/aislamiento & purificación , Electricidad , Electrodos , Luz , Fármacos Fotosensibilizantes/aislamiento & purificación , Hojas de la Planta/química , Raíces de Plantas/química , Energía Solar , Espectrofotometría Ultravioleta , Óxido de Zinc/químicaRESUMEN
AIM: The aim was to study the effect of Aloe vera and clove powder supplementation on carcass characteristics, composition and serum enzymes of Japanese quails. MATERIALS AND METHODS: The study was conducted on 120-day-old Japanese quails, which were randomly divided into four treatment groups viz. T1-control, T2-0.5% A. vera, T3-0.5% clove, and T4-(0.25% A. vera + 0.25% clove) powder, each having three replications consisting of ten Japanese quails. The birds in control group (T1) were fed no supplement whereas in treatments T2-T4 birds were supplemented with A. vera leaf powder, clove, and mixture of both (powdered form) at 0.5, 0.5, and 0.25+0.25% of feed, respectively. After 7 days of brooding, a feeding trial of 35 days was conducted. On 35(th) day of trial birds were sacrificed and dressing percentage, cut up parts, organ weight, processing losses and carcass meat composition of the breast, thigh, and drumstick were recorded. Blood samples were collected on the same day and serum was separated to evaluate serum enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST). RESULTS: It showed significantly (p<0.05) higher values of dressing percentage with and without giblet and breast weight in the T2-T4 group as compared to the T1 group. No significant (p>0.05) differences were observed in giblet weight, other cut up parts and composition of the breast, thigh, and drumstick muscles in all the treatment groups. Serum ALT and AST did not vary significantly among different treatment groups. CONCLUSION: A. vera and clove supplementation improved the dressing percentage and breast weight without adversely affecting the meat composition and serum enzymes. Thus, these can be used as a growth promoter in Japanese quails.
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To evaluate comprehensively the distribution of established risk factors of stroke among Bangladeshi patients. This is an observational study. It involved 8400 stroke patients from different hospitals in Bangladesh over a period of sixteen years. Common established risk factors of stroke e.g. age, sex, family history, hypertension, diabetes, ischemic heart disease, smoking, obesity, dyslipidaemia, alcoholism, use of oral contraceptive pill, lack of fresh fruit consumption etc. were evaluated in these patients through a preformed questionnaire and data were analyzed. Majority of the stroke events occurred after the age of forty (82.3%) and the ischemic stroke (72%) is the most common. Apart from non modifiable risk factors (advancing age, sex, Family history of stroke) hypertension was the most common modifiable risk factor found in stroke patients (57.6%) followed by smoking (44.6%), tobacco use (24.3%), OCP use in female (40% of female stroke), diabetes (23%), ischemic heart disease (17.1%), obesity (10.6%) and dyslipidaemia (5.3%). Lack of fresh fruit consumption and alcoholism were found in some of the patients. Stroke is common after the age of forty. Ischemic events are commonest type of stroke. Hypertension, smoking, tobacco use, diabetes and ischemic heart disease were five most common risk factors of stroke. Outlining the common stroke risk factors in our settings, may help the physicians and care givers in managing this disabling disease properly.
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Accidente Cerebrovascular/etiología , Adulto , Anciano , Estudios Transversales , Femenino , Humanos , Hipertensión/complicaciones , Masculino , Persona de Mediana Edad , Isquemia Miocárdica/complicaciones , Factores de Riesgo , Fumar/efectos adversosRESUMEN
The use of alternative solvents in the iridium-catalysed hydrogen isotope exchange reaction with developing phosphine/NHC Ir(I) complexes has identified reaction media which are more widely applicable and industrially acceptable than the commonly employed chlorinated solvent, dichloromethane. Deuterium incorporation into a variety of substrates has proceeded to deliver high levels of labelling (and regioselectivity) in the presence of low catalyst loadings and over short reaction times. The preparative outputs have been complemented by DFT studies to explore ligand orientation, as well as solvent and substrate binding energies within the catalyst system.
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Química Orgánica/métodos , Medición de Intercambio de Deuterio/métodos , Hidrógeno/química , Iridio/química , Anilidas/química , Catálisis , Conformación Molecular , Niclosamida/química , Solventes/químicaRESUMEN
The chemically treated Labeo rohita scale is used for synthesizing hydroxyapatite (HAp) biomaterials. Thermogravimetric and differential thermal analyses of fish scale materials reveal the different phase changes with temperature and find out the suitable calcination temperatures. The composition and structures of wet ball-milled calcined HAp powders are characterized by Fourier transform infrared spectroscopy, X-ray diffraction, field emission scanning electron microscopy, transmission electron microscopy, energy dispersive X-ray analysis (EDX). The EDX as well as chemical analysis of fish scale-derived apatite materials confirms that the Ca/P ratio is 1.71. The compressive stress, hardness and porosity have been evaluated on sintered HAp biomaterials. The cell attachment on HAp surfaces, cytotoxicity evaluation and MTT assay, which are carried out in RAW macrophage-like cell line media demonstrate good biocompatibility. The histological analysis also supports the bioaffinity of processed HAp biomaterials in Wistar rat model for investigating the contact reaction and stability at the artificial or natural prosthesis interface.
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Materiales Biocompatibles/química , Cyprinidae , Durapatita/química , Andamios del Tejido/química , Animales , Apatitas/química , Línea Celular , Fuerza Compresiva , Macrófagos/metabolismo , Ensayo de Materiales , Ratones , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Porosidad , Polvos , Ratas , Ratas Wistar , Espectroscopía Infrarroja por Transformada de Fourier , Temperatura , Difracción de Rayos XRESUMEN
Abortions and stillbirths were noticed in pregnant goats on a farm in the state of Gujarat, India. About 50% of the pregnant goats aborted or gave birth to dead kids. Bluetongue virus (BTV) antibody in the sera of affected goats was detected using a competitive enzyme-linked immunosorbent assay (ELISA). Viral antigen in the blood of these goats and in the aborted fetal spleens was detected using a sandwich ELISA. Two viruses (SKN-9, SKN-10) were isolated in cell culture from aborted fetal spleens and were confirmed as Orbivirus by demonstration of ten bands in RNA polyacrylamide gel electrophoresis and identified as BTV-1 by sequencing of the VP2 gene. Sequence analyses revealed thatthese isolates were very closely related to a BTV-1 (strain SKN-8) isolated from Culicoides vectors captured on the same farm one month after the occurrence of abortion. Isolation of BTV-1 from fetuses is probably evidence of transplacental transmission of the wild-type strain, because attenuated or laboratory-adapted BTV-1 strains have never been used in this region. This may have important implications in the epidemiology of bluetongue, considering the presence of many BTV serotypes in India.
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Feto Abortado/virología , Aborto Veterinario/virología , Virus de la Lengua Azul/aislamiento & purificación , Lengua Azul/complicaciones , Enfermedades de las Cabras/virología , Animales , Lengua Azul/virología , Virus de la Lengua Azul/clasificación , Virus de la Lengua Azul/genética , Proteínas de la Cápside/genética , Proteínas de la Cápside/metabolismo , Femenino , Regulación Viral de la Expresión Génica , Cabras , Filogenia , Embarazo , Estudios SeroepidemiológicosRESUMEN
Bluetongue virus serotype-1 (BTV-1) was isolated from Culicoides oxystoma vectors captured on livestock farms in two places of Gujarat, India. The viruses were isolated on BHK-21 cells, which produced characteristic BTV-related cytopathic effects between 24 and 48 h post-infection. Virus antigen was demonstrated in infected cells at different passage by a BTV-specific sandwich ELISA. Further, polyacrylamide gel electrophoresis and silver staining of viral genomic RNA revealed ten double-stranded RNA segments characteristic of BTV. Serotype of the isolates was identified by virus neutralization and PCR coupled with sequencing. The isolates were designated as SKN-7 and SKN-8 and their genome segment-2 (VP2) were sequenced. Phylogenetic analyses revealed very close relationship between them although they are not identical. SKN-8 showed closer relationship with a recently isolated BTV-1 from goat. Bluetongue virus was earlier isolated from Culicoides in adjacent state more than 20 years ago, although the serotype of the virus was not determined.