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Herein, we report dye-sensitized solar cells (DSCs) based on conventional nanocrystalline TiO2 photoanodes decorated with one-dimensional (1D) CNT-TiO2 core-shell structures (CTH). The core-shell nanotubes are synthesized by a simple sol-gel template-assisted method via in situ deposition of TiO2 on the surface of non-covalently functionalized CNTs. The core-shell nanotubes are well characterized by various techniques. Field emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM) images show that formation of the TiO2 shell on the surface of the CNT core follows a layer or Frank-van der Merwe growth mode, resulting in a highly uniform interface with excellent charge transfer from the TiO2 conduction band into the CNTs. The thickness and crystal structure of the TiO2 shell can be tailored by controlling the processing parameters. X-ray photoelectron spectroscopy (XPS) and Raman spectroscopy verify that CNTs have no surface defects and are well preserved using the employed method and the subsequent heat treatment in air, respectively. UV-vis spectroscopy and photoluminescence spectroscopy reveal an extension to visible regions with an increase in overall intensity and a significant reduction in charge recombination due to a shift of the Fermi level toward positive potentials. We find an increase by up to 37% in the DSC device's power conversion efficiency by incorporating the CNT-TiO2 core-shell nanotubes into the nanoparticle TiO2 photoanode due to the charge recombination reduction and electron injection enhancement.
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AIM: Previous research indicates that high tie of the inferior mesenteric artery during anterior resection for rectal cancer might be associated with an increased risk of postoperative functional disturbances. The goal of this population-based retrospective cohort study was to further investigate that association. METHOD: Patients who underwent anterior resection for rectal cancer from April 2011 to September 2012 were identified through the Swedish Colorectal Cancer Registry. Bowel and urogenital function were assessed by a postal questionnaire 2 years after surgery. Information on the level of mesenteric tie and clinical variables was retrieved from the registry. The outcome was defined as any defaecatory, urinary or sexual dysfunction as reported by the patient. The association between high tie and the outcome was evaluated with multivariable logistic and linear regression with adjustment for confounders, such as sex, body mass index, comorbidity and preoperative radiation. RESULTS: With a response rate of 86%, 805 patients were included in the study. Of these, 46% were operated with high tie. After adjustment for confounders, high tie did not affect the risk of faecal incontinence (OR 0.85; 95% CI 0.59-1.22), urinary incontinence (OR 0.94; 95% CI 0.63-1.41) or various aspects of sexual dysfunction (erectile dysfunction, anejaculation, dyspareunia and coital vaginal dryness). However, an association between high tie and defaecation at night was detected (OR 1.44; 95% CI 1.02-2.03). CONCLUSION: This study does not support that the level of vascular tie influences the risk of major defaecatory, urinary or sexual disturbances 2 years after anterior resection for rectal cancer.
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Ligadura/efeitos adversos , Complicações Pós-Operatórias/etiologia , Neoplasias Retais/cirurgia , Reto/cirurgia , Idoso , Incontinência Fecal/etiologia , Incontinência Fecal/fisiopatologia , Feminino , Humanos , Ligadura/métodos , Masculino , Artéria Mesentérica Inferior/cirurgia , Pessoa de Meia-Idade , Complicações Pós-Operatórias/fisiopatologia , Período Pós-Operatório , Neoplasias Retais/fisiopatologia , Sistema de Registros , Estudos Retrospectivos , Disfunções Sexuais Fisiológicas/etiologia , Disfunções Sexuais Fisiológicas/fisiopatologia , Suécia , Fatores de Tempo , Incontinência Urinária/etiologia , Incontinência Urinária/fisiopatologiaRESUMO
Background: Heart transplantation is an established treatment for end-stage heart failure patients, but its cost-effectiveness is under question. Objective: This study aimed to assess the cost of heart transplantation in Iran as a developing country in Asia to contribute to future planning in the region. Methods: This study was conducted in two phases. First, in a retrospective multicenter study, hospital data of heart transplant and hospitalization of three active heart transplant centers in Tehran, Iran, were reviewed from April 2013 to May 2015. Then pre-transplantation, transplantation, and one-year posttransplant costs were calculated according to the ABC (activity-based costing) method in 2016. Results: Data were obtained for 120 patients, among which 95 (79.17%) were males with a mean (SD) age of 35.31±13.41 years. Mean (SD) hospital and ICU length of stay were 17.85±14.91 and 9.74±8.94 days, respectively. A significant correlation existed between the mean of hospital and ICU length of stay (P<0.001, r: 0.754). The mean heart transplant and hospitalization cost was 3445.47±1243.29 USD from 2013 to 2015. Using the activity-based costing method, the cost of pre-transplantation, transplantation, and one-year -post-transplantation were extracted 6.5%, 73.5%, and 20%, respectively, with a total cost of 26232 USD. Conclusion: Compared to other countries, the cost of heart transplantation in Iran is very low. Numerous reasons lead to this difference. Firstly, a heart transplantation surgery is performed in university-based hospitals and is supported by the government. On the other hand, a significant difference exists between personnel costs in Iran compared to other countries.
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AIM: Nigella sativa (N. sativa) is a plant widely used in traditional medicine of North African countries. During the last decade, several studies have shown that extracts from the seeds of N. sativa have antidiabetic effects. METHODS: Our group has recently demonstrated that N. sativa seed ethanol extract (NSE) induces an important insulin-like stimulation of glucose uptake in C2C12 skeletal muscle cells and 3T3-L1 adipocytes following an 18 h treatment. The purpose of the present study was to elucidate the pathways mediating this insulin-like effect and the mechanisms through which these pathways are activated. RESULTS: Results from western immunoblot experiments indicate that in C2C12 cells as well as in H4IIE hepatocytes, but not in 3T3-L1 cells, NSE increases activity of Akt, a key mediator of the effects of insulin, and activity of AMP-activated protein kinase (AMPK), a master metabolic regulating enzyme. To test whether the activation of AMPK resulted from a disruption of mitochondrial function, the effects of NSE on oxygen consumption were assessed in isolated liver mitochondria. NSE was found to exhibit potent uncoupling activity. CONCLUSION: Finally, to provide an explanation for the effects of NSE in adipocytes, PPARgamma stimulating activity was tested using a reporter gene assay. Results indicate that NSE behaves as an agonist of PPARgamma. The data supports the ethnobotanical use of N. sativa seed oil as a treatment for diabetes, and suggests potential uses of this product, or compounds derived thereof, against obesity and the metabolic syndrome.
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Proteínas Quinases Ativadas por AMP/efeitos dos fármacos , Adipócitos/metabolismo , Hipoglicemiantes/farmacologia , Músculo Esquelético/metabolismo , Nigella sativa/química , Extratos Vegetais/farmacologia , Proteínas Quinases Ativadas por AMP/metabolismo , Adipócitos/efeitos dos fármacos , Animais , Western Blotting , Células Cultivadas , Humanos , Músculo Esquelético/efeitos dos fármacos , PPAR gama/metabolismo , Extratos Vegetais/química , Sementes/química , Transdução de SinaisRESUMO
PURPOSE: We evaluated the expression and activation of Notch pathway genes in the adult human and murine corneal epithelium during proliferation. METHODS: The expression of Notch pathway genes in the limbal and central human corneal epithelium was compared by reverse transcription polymerase chain reaction (RT-PCR). Their expression pattern was examined by immunofluorescence and in situ hybridization. The temporal expression of Notch1 during murine wound healing was assessed by RT-PCR. Notch activity was determined using western blot for the Notch intracellular domain (NotchIC). The expression of Hes1 was evaluated in cell culture. RESULTS: The expression of Notch1 and Jagged1 was higher in the human limbal epithelium while the expression of Hes1 and Hes5 was higher in the central cornea. Expression of Notch1, Jagged1, and Hes1 was found predominantly in the basal and immediate suprabasal cells. During neonatal corneal development, NotchIC was detected in occasional cells at P10 while at P15 and P90, it was found in the basal and early suprabasal layers. NotchIC was found to be lower in the limbal compared to central corneal epithelium. The expression of Notch1 was lower at 24 h post-wounding but was completely restored in six days. The levels of NotchIC were decreased at 24 h post-wounding and after application of topical phorbol myristate. In vitro, the expression of Hes1 was higher in confluent cells maintained under high calcium conditions. CONCLUSIONS: The inverse correlation between Notch signaling and the proliferative status of the corneal epithelium is consistent with the idea that Notch plays a role in corneal epithelial differentiation.
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Regulação para Baixo/genética , Epitélio Corneano/citologia , Receptores Notch/genética , Transdução de Sinais , Adulto , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Proliferação de Células , Células Cultivadas , Epitélio Corneano/embriologia , Epitélio Corneano/patologia , Imunofluorescência , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Hibridização In Situ , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Proteína Jagged-1 , Limbo da Córnea/citologia , Limbo da Córnea/embriologia , Limbo da Córnea/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Pessoa de Meia-Idade , Modelos Animais , Estrutura Terciária de Proteína , Receptores Notch/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas Serrate-Jagged , Fatores de Transcrição HES-1RESUMO
BACKGROUND: Tuberculosis (TB) is an important cause of morbidity and mortality in renal transplant recipients and, because of its infrequency and the lack of medical awareness, it is usually misdiagnosed. This study was carried out to determine frequency and weight of multiple risk factors for post kidney transplantation TB. METHODS: A total of 44 cases (0.3%), out of 12,820 patients from 12 major kidney transplantation centers in Iran from 1984 to 2003, were compared with 184 healthy transplant subjects who were transplanted by the same surgical team. RESULTS: The mean age of cases and controls was 37.7 (13-63) and 35.6 (8-67) years (P=0.3), respectively. The mean duration of pre-transplantation hemodialysis was 30.3 (3-168) months in cases and 18.2 (1-180) months in controls (P=0.03). A positive past history of TB was detected in 2 cases and 1 control (P=0.3). The mean doses of initial and maintenance immunosuppressive drugs in cases and controls were not significantly different. A total of 25 cases (56.8%) and 60 controls (32.6%) had rejection before diagnosis of TB (P=0.004; OR=2.7, CI(95%): 1.3-5.6). CONCLUSIONS: To our knowledge, this is the first study that demonstrated an increase in the risk of post-transplant TB by increasing the duration of pre-transplant hemodialysis and the number of post-transplant rejection episodes as 2 immunocompromised states. Further study is needed to clarify our new findings, specifically in relation to different immunosuppressive regimens.
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Transplante de Rim/efeitos adversos , Tuberculose Pulmonar/epidemiologia , Tuberculose/epidemiologia , Adolescente , Adulto , Estudos de Casos e Controles , Feminino , Rejeição de Enxerto , Humanos , Irã (Geográfico)/epidemiologia , Masculino , Pessoa de Meia-Idade , Diálise Renal , Fatores de Risco , Transplante Homólogo/efeitos adversos , Tuberculose/microbiologia , Tuberculose Pulmonar/microbiologiaRESUMO
BACKGROUND: The health system faced a new problem of an increasing number of civilian victims of landmine explosions at the end of Iran-Iraq war. METHODS: In a descriptive survey from 1998 to 2004, data was collected retrospectively from medical records of Shahid Motahhary Hospital, Urmia University of Medical Sciences, Iran. RESULT: The study covered 156 victims. 80% of the casualties were civilians of which 95% were male. Injuries led to amputation in 73.3% of the victims. The mortality rate was 3.8%. Blood transfusions was given to 93 (62%) victims. There were 52.6% pattern I, 14.6% pattern II and 32.6% pattern III injuries according to International Committee of the Red Cross (ICRC) classification. CONCLUSION: Mine awareness programs should be conducted amongst civilians who live in high-risk areas. Improved health infrastructure with trained personals for emergency care and early transfer of the casualties would reduce morbidity and mortality. Studies are required to understand the social and public health consequences of this problem.
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A 30-year old male right handed professional tennis player complained about reduced athletic performance, chronic pain and restricted extension of his right wrist. Lateral radiograph of the right wrist demonstrated an osteophyte projecting from the dorsal lip of the lunate bone. The presence of an osteophyte on the lateral radiograph of the lunate along with the history, clinical examination, intra-operative findings, and post-operative satisfactory result made the diagnosis of dorsal luno-capitate impingement syndrome reasonable.
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gp100 is a melanocyte lineage-specific antigen recognized by tumor-infiltrating lymphocytes whose adoptive transfer has been associated with tumor regression in patients with metastatic melanoma. The peripheral blood mononuclear cells of five melanoma patients were sensitized in vitro with synthetic peptides to elicit antigen-specific cytotoxic T lymphocyte (CTL) lines against four gp100 epitopes. These epitope-specific CTL lines were generated following weekly in vitro stimulation with the synthetic decamer G10(476) (V-L-Y-R-Y-G-S-F-S-V) or the nonamers G9(280) (Y-L-E-P-G-P-V-T-A), G9(154) (K-T-W-G-Q-Y-W-Q-V), or G9(209) (I-T-D-Q-V-P-F-S-V) pulsed onto autologous irradiated peripheral blood mononuclear cells. These lines grew as long as 4 months in culture in low-dose interleukin 2 (30 IU/ml) and exhibited antigen-specific, MHC class I-restricted lysis of peptide-pulsed T2 cells and HLA-A2+, gp100+ established melanoma cell lines. G10(476)- and G9(280)-specific CTLs demonstrated specific release of granulocyte-macrophage-colony-stimulating factor and tumor necrosis factor alpha in response to T2 cells pulsed with relevant peptide, as well as to gp100+ melanoma cell lines. These results demonstrate that several peptides derived from the gp100 protein are presented on the surface of melanoma cells and are sufficiently immunogenic to generate, in vitro, potent CTLs capable of cytolysis and the secretion of cytokines. Therefore, for HLA-A2+ melanoma patients, these and possibly other gp100 peptides could represent good candidates for antigen-specific immunotherapy either singly or in a multivalent regimen.
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Antígenos de Neoplasias/imunologia , Epitopos/imunologia , Ativação Linfocitária/efeitos dos fármacos , Glicoproteínas de Membrana/imunologia , Proteínas de Neoplasias/imunologia , Peptídeos/farmacologia , Linfócitos T Citotóxicos/imunologia , Sequência de Aminoácidos , Antígenos de Neoplasias/farmacologia , Divisão Celular/efeitos dos fármacos , Divisão Celular/fisiologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Antígeno HLA-A2/imunologia , Humanos , Imunoterapia Adotiva , Cinética , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Ativação Linfocitária/imunologia , Melanoma/sangue , Melanoma/imunologia , Melanoma/terapia , Glicoproteínas de Membrana/farmacologia , Dados de Sequência Molecular , Proteínas de Neoplasias/farmacologia , Sensibilidade e Especificidade , Linfócitos T Citotóxicos/efeitos dos fármacos , Linfócitos T Citotóxicos/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Antígeno gp100 de MelanomaRESUMO
Primary immunodeficiency disorders include a variety of diseases that render patients more susceptible to infections. To determine the percentage of different primary immunodeficiency disorders diagnosed in the Children's Medical Center Hospital affiliated to Tehran University of Medical Sciences in Iran, we retrospectively reviewed the charts of the patients being referred to our hospital for immunologic evaluation of recurrent infections during a 20 year period. Among these patients, antibody deficiencies were the most frequent ones and were found in 52.6% of patients (n = 130). T-cell disorders, phagocytic disorders and complement deficiencies were found to be present in 24.69% (n = 61). 22.2% (n = 55) and 0.4% (n = 1) respectively. On the whole, common variable immunodeficiency was the most frequent disorder (n = 65), followed by ataxia telangiectasia (n = 39), X-linked agammaglobulinemia (n = 33), chronic granulomatous disease (n = 29) and selective IgA deficiency (n = 20). This study reveals that antibody deficiencies are the most common type of disorders as shown in other studies. A comparative study shows some differences between our results and other registries. This article also indicates that immunodeficiency disorders should be considered in patients with recurrent infections.
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Síndromes de Imunodeficiência/diagnóstico , Síndromes de Imunodeficiência/epidemiologia , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Síndromes de Imunodeficiência/classificação , Síndromes de Imunodeficiência/genética , Lactente , Irã (Geográfico)/epidemiologia , Masculino , Fenótipo , Estudos RetrospectivosRESUMO
This study was conducted to determine whether interferon-alpha-2b (IFN-alpha-2b) can be encapsulated in liposomes without compromising its anti-fibrogenic effects on human dermal fibroblasts. The rationale for this approach is that systemic administration of IFN-alpha-2b by injection for treatment of dermal fibrosis is uncomfortable, requires a large quantity of the cytokine, and cannot be easily used in children. Liposomes are potentially useful as vehicles for the topical delivery of drugs if they can be encapsulated without loss of biologic activity. Empty sonicated vesicles composed of dioleoyl-phosphatidylcholine:dioleoyl-phosphatidylglycerol at a molar ratio of 7:3 were mixed with various concentrations of IFN-alpha-2b and then dried and rehydrated. An enzyme-linked immunosorbent assay (ELISA) was used to determine the efficiency of encapsulation and the stability of the preparation under experimental conditions. Greater than 80% of added IFN-alpha-2b became associated with the liposomes and remained encapsulated for up to 5 d at 4 degrees C. The rate of release increased markedly at 37 degrees C. Liposome-encapsulated IFN-alpha-2b (2000 units per ml) significantly reduced the proliferation of dermal fibroblasts (60 +/- 8.8 vs. 100 +/- 8, mean +/- SEM, p < 0.05, n = 8) and the levels of mRNA for type I (41.5 +/- 8.7% vs 100 +/- 18, p < 0.05, n = 4) and type III (68 +/- 8.4% vs 100 +/- 4.9%, p < 0.05, n = 3) procollagen, as analyzed on northern blots. This was consistent with the reduction found in collagen in conditioned medium from treated fibroblasts. In contrast, treatment increased levels of mRNA for collagenase (241 +/- 42% vs 100 +/- 3.4, p < 0.05, n = 3) and collagenase activity (289 +/- 5.8% vs 100 +/- 10.9%, p < 0.05, n = 9) in conditioned medium. This last effect was probably not due to a reduction in TIMP-1 (tissue inhibitor of metalloproteinase-1) because levels of mRNA for this inhibitor were not lower in treated cells. The efficacy of liposome-associated IFN-alpha-2b in vitro supports the concept of the topical use of this anti-fibrogenic agent for treatment of fibroproliferative disorders.
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Fibroblastos/efeitos dos fármacos , Interferon-alfa/administração & dosagem , Pele/citologia , Pele/patologia , Divisão Celular/efeitos dos fármacos , Colagenases/genética , Colagenases/metabolismo , Portadores de Fármacos , Estabilidade de Medicamentos , Fibroblastos/citologia , Fibrose/prevenção & controle , Glicoproteínas/genética , Humanos , Interferon alfa-2 , Lipossomos , Inibidores de Metaloproteinases de Matriz , Pró-Colágeno/genética , Inibidores de Proteases/metabolismo , RNA Mensageiro/metabolismo , Proteínas Recombinantes , Fatores de Tempo , Inibidores Teciduais de MetaloproteinasesRESUMO
The full length sequence for the human pathogen coxsackievirus B6 (CVB6, Schmitt strain) has been determined. We used long RT-PCR to generate full length DNA amplicon of CVB6, and then directly sequenced the amplicons. One-step cloning of the full length amplicon enabled us to obtain an infectious clone of CVB6. RNA generated from CVB6 amplicon DNA or CVB6 clones, by transcription with T7 RNA polymerase, was demonstrated to be infectious upon transfection into HeLa cells in vitro. The CVB6 genome is characteristic of enteroviruses, with a 5'-non-translated region (743 nucleotides) followed by an open reading frame (encoding a 2184 amino acid polyprotein) and a 3'-non-translated region (100 nucleotides) and polyadenylated tail. The predicted amino acid sequence of CVB6 clustered with the other CVB serotypes and swine vesicular disease virus (SVDV).
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Enterovirus Humano B/genética , Regiões 3' não Traduzidas/genética , Regiões 5' não Traduzidas/genética , Sequência de Bases , Clonagem Molecular/métodos , Sequência Consenso , Primers do DNA , Enterovirus/classificação , Enterovirus/genética , Enterovirus Humano B/classificação , Células HeLa , Humanos , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , TransfecçãoRESUMO
Rapid, sensitive peroxidase labelled antibody (PLA) assays using microtiter systems, were developed for detection of hog cholera virus (HCV) and cross-reacting bovine viral diarrhea virus (BVDV) antibodies in pig sera. HCV-infected pig kidney cell line (PK 15) prepared in microtiter plates were fixed and used in PLA assays. After inoculation with test serum, bound antibodies (HCV/BVDV) were reacted with either horseradish peroxidase (HRP) conjugated anti-porcine immunoglobulin (H & L) or biotinylated protein A (BPA) and subsequent HRP labelled avidin (A). Positive reactions were easily visualized under an inverted light microscope as foci of brown colored cells after enzyme degradation of hydrogen peroxidase in the presence of amino-ethylcarbazole (AEC). The PLA assays were superior to the indirect fluorescent antibody (IFA) test in detecting anti-HCV antibodies in porcine sera collected early after inoculation of pigs with a lapinized HCV vaccine. The performances of the PLA, IFA and FA neutralization (FAN) tests in measuring the immune response in the vaccinated pigs were comparable. Cross-reacting anti-BVDV antibody, as measured by a microtiter serum neutralization (MTSN) test, was not demonstrable in vaccinated pigs until they were challenged with a virulent HCV, 13 weeks later. The PLA assays relative to the IFA test detected more reactive samples among porcine field sera collected from HC-free pigs in Canada. Of 795 samples, 24 (3.01%) were reactive in the PLA employing HRP anti-porcine IgG, and 21 (2.6%) in the PLA, using BPA-HRP-A. When 324 of these sera were screened by the IFA test (using HC antigen), only one sample (0.30%) was found reactive.(ABSTRACT TRUNCATED AT 250 WORDS)
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Anticorpos Antivirais/análise , Vírus da Febre Suína Clássica/imunologia , Vírus da Diarreia Viral Bovina/imunologia , Técnicas Imunoenzimáticas , Imunoglobulina G/análise , Pestivirus/imunologia , Animais , Anticorpos Antivirais/biossíntese , Doença das Mucosas por Vírus da Diarreia Viral Bovina/diagnóstico , Bovinos , Linhagem Celular , Peste Suína Clássica/diagnóstico , Reações Cruzadas , Imunofluorescência , Imunoglobulina G/imunologia , Testes de Neutralização , Sensibilidade e Especificidade , Testes Sorológicos , Suínos/imunologia , Suínos/microbiologia , Vacinas Virais/administração & dosagemRESUMO
Anti-idiotypic antibodies (anti-Id or Ab2) were generated in Balb/c mice against either mouse monoclonal or swine polyclonal antibodies (Ab1) to pseudorabies virus (PRV) antigens by conventional and sequential immunization methods. In the conventional method, one antibody preparation was repeatedly injected into the animals, whereas three anti-PRV antibody preparations were used alternately for the sequential immunization procedure. Anti-Ids were serologically characterized for possession of the Ab2s that detect shared idiotype (IdX) on antibodies to PRV antigens. Only the Ab2s that were generated by the sequential immunization method recognized the IdX present on murine and swine antibodies to PRV. The sequential immunization method described herein was anticipated to be helpful for generating virus specific Ab2s as candidates for serodiagnostic reagents or vaccines.
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Anticorpos Anti-Idiotípicos , Antígenos Virais , Herpesvirus Suídeo 1/imunologia , Animais , Anticorpos Anti-Idiotípicos/biossíntese , Anticorpos Monoclonais , Anticorpos Antivirais , Especificidade de Anticorpos , Ensaio de Imunoadsorção Enzimática , Feminino , Imunização , Idiótipos de Imunoglobulinas , Camundongos , Camundongos Endogâmicos BALB C , Pseudorraiva/imunologia , Suínos , Doenças dos Suínos/imunologia , Virologia/métodosRESUMO
The suitability of autoclaved tap water for the preparation of ELISA reagents and washing buffer was compared with that of ultrapure water, in a standard indirect ELISA for the detection of antibodies to pseudorabies virus (PRV). The performance of the assay, using autoclaved tap water (AT-ELISA) compared favourably to that of the standard assay, using ultrapure water (UP-ELISA) in detecting anti-PRV antibodies in sequential serum samples from a pig experimentally infected with PRV. While both the UP-ELISA and AT-ELISA proved reliable in detecting anti-PRV antibodies in a coded proficiency serum panel (n = 60), the AT-ELISA detected fewer positive sera than the UP-ELISA in evaluating a limited number (n = 80) of field samples. The results suggest that autoclaved tap water may be substituted for ultrapure water for the preparation of ELISA reagents when or where ultrapure water may not be available.
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Ensaio de Imunoadsorção Enzimática , Esterilização/métodos , Água , Animais , Anticorpos Antivirais/isolamento & purificação , Herpesvirus Suídeo 1/imunologia , Temperatura Alta , Suínos , Doenças dos Suínos/imunologiaRESUMO
Bluetongue virus (BTV) is an arthropod-borne orbivirus that infects sheep, wild ruminants and occasionally cattle. Detection and specific identification of BTV is a multistep process. The first step involves the isolation of the virus from the animal's blood or other tissues, followed by inoculation of embryonating chicken eggs (ECE). After the virus has been amplified in ECE, it is passaged into BHK-21 cell culture for subsequent replication and identification. The virus is then amplified further and identified in microtiter plates by the immunoperoxidase assay using a group specific monoclonal antibody. Finally, the viral isolate is typed by a virus neutralization test.
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Vírus Bluetongue/isolamento & purificação , Bluetongue/virologia , Animais , Anticorpos Monoclonais , Anticorpos Antivirais , Bluetongue/sangue , Vírus Bluetongue/classificação , Bovinos , Linhagem Celular , Embrião de Galinha , Efeito Citopatogênico Viral , Técnicas Imunoenzimáticas , Testes de Neutralização , Vísceras/virologiaRESUMO
A modified solid phase blocking enzyme immunosorbent assay (ELISA), using a monoclonal antibody (McAb) against the group specific bluetongue virus (BTV) antigen is described for detection of anti-BTV antibodies in cattle and sheep sera. Dots of an optimal dilution of BTV antigens were adsorbed to nitrocellulose (NC) paper (hence dot-ELISA) and then the remaining adsorptive sites were saturated with gelatin. After exposure to bovine or ovine test serum the NC strips were reacted with the McAb. The presence of McAb was detected with a peroxidase-conjugated anti-mouse IgG (H and L). In the absence of anti-BTV antibody in test sera, BTV antigen sites were reactive with McAb as indicated by a brown colored dot after enzyme degradation of hydrogen peroxide in the presence of diamino benzidine (DAB) or amino ethylcarbazole (AEC). In the presence of sufficient anti-BTV antibody no color reaction was observed. The blocking (B) dot-ELISA was superior to the agar gel immunodiffusion (AGID) in detecting anti-BTV antibodies in bovine and ovine sera early after experimental infection with BTV type 10. In 5 of 7 animals inoculated by combined intravenous and subcutaneous routes, anti-BTV antibodies in sera were detectable as early as 7 days post infection (DPI), all of which were AGID negative. Comparable B-dot-ELISA and AGID results were found in 23 paired sera (pre and 20 DPI) from cattle experimentally infected with different types of BTV and in 100 AGID negative sera from Ontario dairy and Alberta beef cattle.(ABSTRACT TRUNCATED AT 250 WORDS)
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Anticorpos Monoclonais , Anticorpos Antivirais/análise , Vírus Bluetongue/imunologia , Ensaio de Imunoadsorção Enzimática , Reoviridae/imunologia , Animais , Anticorpos Monoclonais/imunologia , Antígenos Virais/imunologia , Bovinos/sangue , Bovinos/imunologia , Imunodifusão , Ovinos/sangue , Ovinos/imunologiaRESUMO
A rapid, simple dot immunoperoxidase assay (DIPA) is described for visual detection and identification of bluetongue virus (BTV) antigens in samples of infected cell culture fluid. The assay was performed using nitrocellulose (NC) paper and 'dipsticks'. Dots of samples were adsorbed to the NC surface and the remaining non-specific binding sites were blocked with skim milk solution. BTV was detected with either of two murine monoclonal antibodies (4H4, 5G12) to the major group specific antigens of BTV, and the complex was reacted with a peroxidase conjugated anti-mouse immunoglobulin G (heavy- and light-chain specific). Positive reactions were easily visualized as brown spots after enzyme degradation of substrate containing H2O2 and diaminobenzidine (DAB). The DIPA was specific in detecting BTV in samples of cell culture fluid from baby hamster kidney (BHK-21) cells infected with U.S.A. isolates of the five BTV serotypes (2, 10, 11, 13 and 17) known to exist in the U.S.A., and South African isolates of 17 BTV serotypes (1-12, 14-16, 18 and 20), but not with two North American isolates of epizootic hemorrhagic disease of deer virus (EHDV) representing serotypes 1 and 2. Attempts to detect BTV directly in infected sheep blood cells and chick embryo tissue suspensions by DIPA were unsuccessful. Of 55 cell culture fluid samples examined from BHK-21 or Vero cell monolayers inoculated with 55 clinical specimens, propagated initially in embryonating chicken egg (ECE) 11 proved positive and 44 were negative by DIPA. The results were in complete agreement with the conventional ECE and tissue culture isolation systems. The DIPA appears to have potential application, especially as a 'dipstick' kit, for rapid and inexpensive laboratory diagnosis of bluetongue virus infection.
Assuntos
Antígenos Virais/análise , Vírus Bluetongue/imunologia , Animais , Anticorpos Monoclonais , Bluetongue/imunologia , Células Cultivadas , Embrião de Galinha , Colódio , Técnicas Imunoenzimáticas/veterinária , OvinosRESUMO
A total of 3,370 plasma samples, collected from residents of three mountainous regions, located in Azerbaijan and Khuzestan provinces, north-west and south-west Iran, were tested for Toxoplasma antibodies by the indirect fluorescent antibody technique. The over-all sero-positive rate was 12.8%, with a general increase in positivity with increasing age. The distribution of antibodies to Toxoplasma was found to be different in different ethnic groups in the regions studied. The Toxoplasma sero-positive rate in these mountainous areas was considerably lower than that in the Caspian Sea littoral, in northern Iran, obtained in our previous study.
Assuntos
Anticorpos/análise , Toxoplasmose/epidemiologia , Fatores Etários , Etnicidade , Feminino , Imunofluorescência , Humanos , Irã (Geográfico) , Masculino , Toxoplasmose/imunologiaRESUMO
A blocking (B) dot enzyme-linked immunosorbent assay (ELISA), using a monoclonal antibody (mAb) against a group specific antigen of bluetongue virus (BTV) is described for the detection of BTV antibodies to BTV in cattle sera. Dots of BTV antigens were adsorbed to nitrocellulose (NC) strips and/or NC mounted in the windows of dipsticks. After blocking the remaining sites of the NC paper with milk powder solution and immersion in the test sample, the NC strips and dipsticks were exposed to mAb. Bound mAb was detected with peroxidase conjugated anti-mouse IgG (H and L). In the absence of anti-BTV antibody in the test sample, BTV antigen sites were reactive with mAb as indicated by a brown colored dot in the presence of the enzyme substrate, hydrogen peroxide and diaminobenzidine. In the presence of sufficient anti-BTV antibodies no color reaction was observed. The performance of these assays in detecting anti-BTV antibody in field blood eluate samples, prepared from whole blood dried on filter paper, from 395 bluetongue-free cattle in Canada and 635 sentinel cattle in Florida, USA, was evaluated and compared with the standard competitive (C) ELISA. The specificity of the dipstick B-dot ELISA was identical to that of the C-ELISA in testing of BT-free Canadian cattle but not in the testing of samples from the sentinel cattle in Florida, resulting in values of 100% diagnostic and 88.9% relative specificity, respectively. Based on the C-ELISA, the specificity of the NC strip B-dot ELISA was low and in the same order as that of the dipstick assay.(ABSTRACT TRUNCATED AT 250 WORDS)