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1.
Brasília; s.n; maio 2020.
Non-conventional in Portuguese | LILACS (Americas), BRISA | ID: biblio-1099659

ABSTRACT

INTRODUÇÃO: O coronavírus da Síndrome Respiratória Aguda Grave 2 (abreviado para SARSCoV-2, do inglês Severe Acute Respiratory Syndrome Coronavirus 2), anteriormente conhecido como novo coronavírus (2019-nCoV), é um agente zoonótico recémemergente que surgiu em dezembro de 2019, em Wuhan, China, causando manifestações respiratórias, digestivas e sistêmicas, que se articulam no quadro clínico da doença denominada COVID-19 (do inglês Coronavirus Disease 2019). Ainda não há informações robustas sobre a história natural da doença, tampouco sobre as medidas de efetividade para o manejo clínico dos casos de infecção pelo COVID19, restando ainda muitos detalhes a serem esclarecidos. No entanto, sabe-se que o vírus tem alta transmissibilidade e provoca uma síndrome respiratória aguda que varia de casos leves ­ cerca de 80% ­ a casos muito graves com insuficiência respiratória - entre 5% e 10% dos casos ­, os quais requerem tratamento especializado em unidades de terapia intensiva (UTI). Sua letalidade varia, principalmente, conforme a faixa etária. TECNOLOGIA: Os testes de diagnóstico para a COVID-19 se destacaram na pandemia de coronavírus em andamento como uma ferramenta essencial para rastrear a propagação da doença. Uma ampla gama de testes diagnósticos para o SARS-CoV-2 está disponível comercialmente, alguns dos quais receberam autorizações para uso por várias agências reguladoras nacionais. Com as informações da sequência genética devidamente identificadas, os testes de diagnóstico baseados na detecção da sequência viral por reação em cadeia da polimerase com transcriptase reversa (RT-PCR) ou plataformas de sequenciamento logo se tornaram disponíveis. Isso permitiu a confirmação do diagnóstico e melhores estimativas da atividade da infecção, que vêm aumentando em velocidades alarmantes. Para a detecção mais sensível de SARS-CoV, MERS-CoV e SARS-CoV-2, recomendavam-se a coleta e o teste de amostras respiratórias superiores e inferiores. O diagnóstico de casos suspeitos era confirmado por testes de RNA com RT-PCR em tempo real ou sequenciamento de próxima geração. Foi demonstrado que o RNA viral poderia ser detectado a partir do swab nasal e faríngeo, lavagem broncoalveolar e plasma sanguíneo usando RT-PCR direcionado ao gene do vírus (5). O padrão-ouro para diagnóstico laboratorial da COVID-19 é a reação da transcriptase reversa, seguida de reação em cadeia da polimerase (RT-PCR) para amostras coletadas no trato respiratório superior ou inferior. OBJETIVO: O objetivo deste relatório é analisar a acurácia dos testes diagnósticos registrados na Agência Nacional de Vigilância Sanitária (ANVISA) até a presente data. METODOLOGIA: foi realizada uma busca por diagnósticos para COVID-19 com registros vigentes na ANVISA. Para tal, foram utilizados os termos "COVID 19", SARS e coronavírus no campo de consulta de registro de produtos para saúde no site da Agência (https://consultas.anvisa.gov.br/#/saude/). Os passos para acesso ao banco de dados de produtos diagnósticos na ANVISA são: 1) consulta produtos; 2) consulta a banco de dados; 3) produtos para a saúde e 4) pesquisa de produtos para a saúde registrados. CONCLUSÕES: A ANVISA já avaliou mais de 120 pedidos de registro de produtos para testagens relacionadas à COVID-19 desde o dia 18 de março. A maior parte das petições aguarda complementação de informações por parte das empresas e outras estão sendo analisadas com prioridade. O tempo médio para avaliação dos registros na ANVISA gira em torno de 15 dias. Atualmente, mais da metade dos registros concedidos diz respeito a testes rápidos para anticorpos. Até a presente data, foram registrados 64 testes para diagnóstico da COVID-19, sendo 15 deles moleculares. O teste de polymerase chain reaction em tempo real (RT-PCR) para identificação de SARS-CoV-2 é um teste de elevada sensibilidade e especificidade, ainda que os doentes com maior carga viral possam ter maior probabilidade de um teste positivo. Os testes moleculares baseados em RNA exigem instalações laboratoriais específicas com níveis restritos de biossegurança e técnica. A sensibilidade e especificidade dos testes sorológicos variaram entre os fabricantes. É importante destacar que uma baixa sensibilidade do teste diagnóstico pode resultar em uma maior probabilidade de detectar falsos-negativos, o que poderia interferir principalmente em casos de indivíduos assintomáticos. Em geral, a sensibilidade dos testes foi superior a 85% e a especificidade, superior a 94%. Os testes sorológicos medem a quantidade de dois anticorpos (IgG e IgM) que o organismo produz quando entra em contato com um invasor. Contudo, o desenvolvimento da resposta de um anticorpo à infecção pode ser dependente do hospedeiro e levar tempo. No caso de SARS-CoV-2, estudos iniciais sugerem que a maioria dos pacientes se converte entre 7 e 11 dias após a exposição ao vírus, embora alguns pacientes possam desenvolver anticorpos mais cedo. Devido a esse atraso natural, o teste de anticorpos pode não ser útil no cenário de uma doença aguda (11). Os testes de anticorpos para SARS-CoV-2 podem facilitar (i) o rastreamento de contatos (os testes baseados em RNA também podem ajudar); (ii) a vigilância sorológica nos níveis local, regional, estadual e nacional; e (iii) a identificação de quem já teve contato com o vírus e, portanto, pode (se houver imunidade protetora) ser imune (11,12). Alguns conjuntos de reagentes para testes sorológicos foram autorizados pela ANVISA em caráter emergencial devido à gravidade da situação e à necessidade de ampliar a testagem da população, mas a validação desses reagentes pelos laboratórios é fundamental, uma vez que poucos trabalhos conseguiram ser publicados até o momento. As aprovações estão de acordo com a Resolução da Diretoria Colegiada (RDC) 348/2020, que define os critérios e os procedimentos extraordinários e temporários para tratamento de petições de registro de medicamentos, produtos biológicos e produtos para diagnóstico in vitro, e mudança pós-registro de medicamentos e produtos biológicos em virtude da emergência de saúde pública internacional decorrente do novo coronavírus. Na RDC, para registro de testes diagnósticos, a ausência de qualquer estudo de desempenho ou restrição de dados deve ser justificada por motivações técnicas que permitam a avalição da confiabilidade dos resultados e da efetividade diagnóstica do produto. Os registros concedidos nas condições dessa Resolução terão a validade de um ano, exceto para situações em que a avaliação da estabilidade seja apresentada por comparação com produtos similares e os demais critérios descritos no Regulamento sejam atendidos. Nesse caso, poderão ter a concessão regular de validade de registro de produtos para saúde por um período de 10 anos. Em resumo, as duas categorias de testes para SARS-CoV-2 podem ser úteis nesse surto, pois, eventualmente, a coleta de múltiplas amostras, regiões e em tempos diferentes durante a evolução da doença pode ser necessária para o diagnóstico da COVID-19.


Subject(s)
Humans , Enzyme-Linked Immunosorbent Assay/instrumentation , Chromatography, Affinity/instrumentation , Fluorescent Antibody Technique/instrumentation , Coronavirus Infections/diagnosis , Reverse Transcriptase Polymerase Chain Reaction/instrumentation , Technology Assessment, Biomedical , Health Evaluation
3.
Electron. j. biotechnol ; 44: 6-13, Mar. 2020. tab, graf, ilus
Article in English | LILACS (Americas) | ID: biblio-1087627

ABSTRACT

BACKGROUND: Hot start can greatly improve specificity, sensitivity and yield of PCR. Non-specific amplification can occur in PCR when reaction mixture is prepared at room temperature, because Taq DNA polymerase is active and the primers can hybridize non-specifically. Hot start Taq DNA polymerases remain inactive at room temperature and are activated after heating at 95°C preventing non-specific amplification. Monoclonal antibodies against Taq DNA polymerase is the first line of reagents used for turn on regular Taq DNA polymerase into Hot start one. The goal of this research was to produce and evaluate Hot Start antibodies derived from chicken eggs. RESULTS: We performed affinity purification of yolk immunoglobulin (IgY) and obtained polyclonal Hot Start antibodies. The yield of specific antibodies was 0.36 mg per egg or 0.2% of total yolk antibodies. The protocol for real time measurement and Hot start IgY activity assessment was developed. We found that Hot start IgY can reversibly block Taq DNA polymerase activity at 50°C and have no negative impact neither on the Taq DNA polymerase activity after denaturation nor on the reverse transcriptase. We estimated that 1.0 µg of Hot start IgY effectively blocks 5 U activity of Taq DNA polymerase. CONCLUSIONS: Egg derived Hot Start polyclonal antibodies are the cheapest source of Hot start antibodies, from one immune egg we can isolate 0.36 mg IgY, this quantity is enough for producing 1800 U activity of Hot start Taq DNA Polymerase.


Subject(s)
Egg Yolk/metabolism , Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/immunology , Temperature , Immunoglobulins/isolation & purification , Immunoglobulins/biosynthesis , Immunoglobulins/immunology , Enzyme-Linked Immunosorbent Assay , Polymerase Chain Reaction , Taq Polymerase , Egg Yolk/immunology , Antibodies, Monoclonal/isolation & purification
4.
Arq. bras. med. vet. zootec. (Online) ; 72(1): 277-280, Jan.-Feb. 2020. tab
Article in English | LILACS (Americas), VETINDEX | ID: biblio-1088939

ABSTRACT

O objetivo do presente estudo foi investigar a soroprevalência de infecção com Toxoplasma gondii (T. gondii) em cabras em cinco províncias do noroeste da China. Soroprevalência foi determinado usando o kit de teste de ensaio de imunoabsorção enzimática (ELISA). A soroprevalência geral foi 21.23% (197/928). Análise estatística revlou que diferenças significativas foram observadas em fêmeas (P= 0.048, OR= 0.567, 95% CI= 0.309 a 1.041) e nos grupos ≥ 2 (P= 0.002, OR= 0.330, 95% CI= 0.224 a 0.488). Nenhuma diferença estatisticamente significativa foi observada entre diferentes províncias. Nossos resultados indicam que a infecção com T. gondii, que pode ter implicações importantes sobre a saúde pública, teve diferenças significativas em sexo e idade, mas nenhuma significância foi observada em diferentes regiões. Além disto, nossos resultados também indicam que infecção por T. gondii em cabras é generalizada nas cinco províncias do noroeste.(AU)


Subject(s)
Animals , Toxoplasma/isolation & purification , Goats/microbiology , Toxoplasmosis, Animal/epidemiology , Enzyme-Linked Immunosorbent Assay/veterinary , China
5.
Arq. bras. med. vet. zootec. (Online) ; 72(1): 1-8, Jan.-Feb. 2020. tab
Article in English | LILACS (Americas), VETINDEX | ID: biblio-1088931

ABSTRACT

Bovine tuberculosis is a worldwide spread zoonotic disease. Intradermal tuberculinizations are the most used diagnostic tests in the world. Serological tests can be an ancillary diagnosis for bovine tuberculosis. The objective of this study was to evaluate the diagnostic performance of the ELISA Mycobacterium Bovis Antibody Test Kit IDEXX ™ in infected herds, which were in different disease control stages. One hundred and twenty animals from two dairy herds of Minas Gerais state, Brazil, were subjected to the ELISA serological test and the comparative cervical tuberculin test (CCT). Diagnostic test parameters were estimated using Bayesian latent class models and concordance between tests estimated by the frequentist approach. The ELISA test presented lower sensitivity than CCT in both herds. Its sensitivity was higher in the herd in sanitation process. Specificity estimates were above 95% in both herds. Kappa index indicated low concordance or even disagreement between tests. According to the results, the ELISA IDEXX should not be used as substitution for CCT. The tests must not be associated in series. Parallel association increased diagnostic sensitivity in the herd which was in the process of sanitation.(AU)


A tuberculose bovina é uma zoonose de distribuição mundial cujos testes mais utilizados para o diagnóstico são as tuberculinizações intradérmicas, simples e compartivas. Contudo, testes sorológicos podem constituir diagnósticos auxiliares. O objetivo deste estudo foi avaliar o desempenho diagnóstico do teste ELISA Mycobacterium Bovis Antibody Test Kit IDEXX ® em rebanhos bovinos infectados, que se encontravam em diferentes estágios de controle da doença. Cento e vinte animais de dois rebanhos leiteiros provenientes do estado de Minas Geais-Brasil foram submetidos ao ELISA e à tuberculinização cervical compartiva (TCC). Avaliou-se o desempenho dos testes por meio de modelos Bayesianos de classe latente e a concordância entre os eles, por meio de estatística frequentista. Uma maior sensibilidade do teste foi observada no rebanho previamente tuberculinizado. Em ambos os rebanhos o TCC foi mais sensível que o ELISA. Especificidade acima de 95% foi encontrada em ambos os rebanhos. Foram observadas baixa concordância ou mesmo discordância entre os testes. De acordo com os resultados obtidos, o teste ELISA-IDEXX não deve ser utilizado em substituição à TCC, tampouco devem ser associados em série. Houve aumento da sensibilidade quando os testes foram associados em paralelo no rebanho que já se encontrava em processo de saneamento.(AU)


Subject(s)
Animals , Cattle , Tuberculosis, Bovine/diagnosis , Enzyme-Linked Immunosorbent Assay/veterinary , Communicable Disease Control/methods , Diagnostic Test Approval
6.
Article in Korean | WPRIM (Western Pacific) | ID: wprim-816641

ABSTRACT

Porcine respiratory disease complex (PRDC) continues to be a significant economic problem to the swine industry. Porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), and Mycoplasma hyopneumoniae (MH) are considered to be the most important pathogens that cause PRDC. In this study, we investigated the prevalence of antibodies against PRRSV and MH in the serum of sows and piglets from 89 domestic commercial pig farms by ELISA, and the presence of viral nucleic acids of PRRSV, including North American and European PRRS, and PCV2 was also investigated in the serum of sows and piglets from 89 domestic commercial pig farms by real-time PCR. In case of PRRSV, 78.7% (70/89) of sows were positive for PRRSV antibody, and 96.6% (86/89) of piglets were positive for PRRSV antibody. For MH, 76.4% (68/89) of sows showed positive for MH antibody. In the PRRSV viral nucleic acid detection experiment, 36.0% (32/89) of sows were positive for PRRSV nucleic acids, and virus nucleic acid was detected in 83.1% (74/89) of piglets. In case of virus type, both North American and European types were detected. In case of PCV2, 15.7% (14/89) of sows were positive for PCV2 nucleic acids. Conclusively, PCV2, PRRSV, and MH were widely distributed in pig farms in Korea. These prevalence data related with PRDC provides clinical information for vaccination strategy and development for the control of PRDC.


Subject(s)
Agriculture , Antibodies , Circovirus , Enzyme-Linked Immunosorbent Assay , Korea , Mycoplasma hyopneumoniae , Mycoplasma , Nucleic Acids , Porcine Reproductive and Respiratory Syndrome , Porcine respiratory and reproductive syndrome virus , Prevalence , Real-Time Polymerase Chain Reaction , Swine , Vaccination
7.
Article in English | WPRIM (Western Pacific) | ID: wprim-816640

ABSTRACT

Canine adenovirus type 1 (CAV-1) causes infectious hepatitis in members of the family Canidae, including dogs. An indirect enzyme-linked immunosorbent assay (I-ELISA) that detects CAV-1 antibodies is required for large-throughput tests of dog sera. We collected 165 serum samples from dogs of Chungbuk and Gyeongbuk provinces between February 2016 and October 2018. The Korean CAV-1 vaccine strain CAV1V was propagated in Madin-Darby canine kidney (MDCK) cells and purified via Nuvia cPrime anion-exchange chromatography; the virus served as an I-ELISA antigen. Virus-neutralizing anti-CAV-1 titers in dog sera were measured using the virus neutralization (VN) method. The I-ELISA was optimized using purified CAV-1 antigen and serum samples. This kit was used to evaluate dog sera. The VN and I-ELISA data were compared. The sensitivity, specificity, and accuracy of the I-ELISA were 97.0%, 74.2%, and 92.7% compared to the VN assay, respectively. The I-ELISA data significantly correlated with those of VN (r = 0.88). These results suggest that the I-ELISA is useful for serosurveillance of CAV-1 in dog sera.


Subject(s)
Adenoviruses, Canine , Animals , Antibodies , Canidae , Chromatography , Dogs , Enzyme-Linked Immunosorbent Assay , Hepatitis A , Humans , Kidney , Methods , Sensitivity and Specificity
8.
Article in English | WPRIM (Western Pacific) | ID: wprim-816639

ABSTRACT

Mayaro virus (MAYV) is a mosquito-transmitted alphavirus that produces an acute, usually non-fatal, febrile illness including Mayaro fever. Like other alphaviruses, the MAYV E1 and E2 envelope glycoproteins are major viral surface antigens that play a key role in host recognition and infection. Here, we report expression and purification methods for recombinant MAYV E1 (rE1) and rE2 using a baculovirus system. Enzyme-linked immunosorbent assays (ELISA) revealed that rE1 and rE2 were antigenic and reacted with human anti-MAYV IgG and IgM. Cross-reactivity was also confirmed with human anti-Chikungunya virus (CHIKV) IgG and IgM. Furthermore, we developed an immunochromatographic strip test (IST) with rE2 to diagnose MAYV infection. Thus, purified rE2 may be valuable tool for rapidly diagnosing MAYV infection.


Subject(s)
Alphavirus , Antigens, Surface , Baculoviridae , Enzyme-Linked Immunosorbent Assay , Fever , Glycoproteins , Humans , Immunoglobulin G , Immunoglobulin M
9.
Article in English | WPRIM (Western Pacific) | ID: wprim-816615

ABSTRACT

BACKGROUND: Acromegaly is a rare disease primarily caused by growth hormone (GH)-secreting pituitary adenomas, and its treatment is costly. Moreover, some patients are unresponsive to treatment. Hence, there are increasing efforts to develop new drugs with improved effectiveness for this disease. BIM23B065 is a novel chimeric molecule that acts on both somatostatin and dopamine receptors. This study aimed to investigate the effects of BIM23B065 compared with those of a somatostatin receptor analog and a dopamine agonist.METHODS: The effects of BIM23B065 on the proliferation, GH and insulin-like growth factor-1 (IGF-1) levels, and extracellular signal-regulated kinase (ERK) 1/2 and cyclic AMP response element binding (CREB) phosphorylation of GH3 cells were investigated with MTS assay, enzyme-linked immunosorbent assay, and Western blotting, respectively. The dosage and treatment duration of BIM23B065 were tested in animal models of GH-secreting pituitary adenoma. The effect of BIM23B065 (3 mg/kg/day) on changes in IGF-1 levels before and after treatment was further investigated.RESULTS: In vitro, BIM23B065 treatment decreased GH release in the culture media and downregulated ERK 1/2 and CREB phosphorylation to 22% and 26%, respectively. In vivo, IGF-1 expression decreased to 50 % after 4 weeks of treatment with BIM23B065 using an osmotic pump implant. Moreover, magnetic resonance imaging results showed that the tumor size decreased significantly following treatment with BIM23B065 for 4 weeks.CONCLUSION: The novel chimeric molecule was effective in decreasing IGF-1 and GH levels and may serve as an effective therapeutic agent for acromegaly.


Subject(s)
Acromegaly , Blotting, Western , Culture Media , Cyclic AMP , Dopamine Agonists , Dopamine , Enzyme-Linked Immunosorbent Assay , Growth Hormone , Growth Hormone-Secreting Pituitary Adenoma , Humans , In Vitro Techniques , Insulin-Like Growth Factor I , Magnetic Resonance Imaging , Models, Animal , Phosphorylation , Phosphotransferases , Pituitary Neoplasms , Rare Diseases , Receptors, Dopamine , Receptors, Somatostatin , Response Elements , Somatostatin
10.
Article in English | WPRIM (Western Pacific) | ID: wprim-762183

ABSTRACT

PURPOSE: The effect of air pollution-related particulate matter (PM) on epithelial barrier function and tight junction (TJ) expression in human nasal mucosa has not been studied to date. This study therefore aimed to assess the direct impact of PM with an aerodynamic diameter less than 2.5 μm (PM2.5) on the barrier function and TJ molecular expression of human nasal epithelial cells. METHODS: Air-liquid interface cultures were established with epithelial cells derived from noninflammatory nasal mucosal tissue collected from patients undergoing paranasal sinus surgery. Confluent cultures were exposed to 50 or 100 µg/mL PM2.5 for up to 72 hours, and assessed for 1) epithelial barrier integrity as measured by transepithelial resistance (TER) and permeability of fluorescein isothiocyanate (FITC) 4 kDa; 2) expression of TJs using real-time quantitative polymerase chain reaction and immunofluorescence staining, and 3) proinflammatory cytokines by luminometric bead array or enzyme-linked immunosorbent assay. RESULTS: Compared to control medium, 50 and/or 100 µg/mL PM2.5-treatment 1) significantly decreased TER and increased FITC permeability, which could not be restored by budesonide pretreatment; 2) significantly decreased the expression of claudin-1 messenger RNA, claudin-1, occludin and ZO-1 protein; and 3) significantly increased production of the cytokines interleukin-8, TIMP metallopeptidase inhibitor 1 and thymic stromal lymphopoietin. CONCLUSIONS: Exposure to PM2.5 may lead to loss of barrier function in human nasal epithelium through decreased expression of TJ proteins and increased release of proinflammatory cytokines. These results suggest an important mechanism of susceptibility to rhinitis and rhinosinusitis in highly PM2.5-polluted areas.


Subject(s)
Asthma , Budesonide , Claudin-1 , Cytokines , Enzyme-Linked Immunosorbent Assay , Epithelial Cells , Fluorescein , Fluorescein-5-isothiocyanate , Fluorescent Antibody Technique , Humans , Interleukin-8 , Mucous Membrane , Nasal Mucosa , Occludin , Particulate Matter , Permeability , Polymerase Chain Reaction , Rhinitis , RNA, Messenger , Tight Junctions
11.
Article in English | WPRIM (Western Pacific) | ID: wprim-782519

ABSTRACT

Currently, the optimal resuscitation fluid remains debatable. Therefore, in the present study, we designed a trometamol-balanced solution (TBS) for use as a resuscitation fluid for hemorrhagic shock. Hemorrhagic shock was induced in 18 male Wistar-Kyoto rats, which were assigned to normal saline (NS), Ringer's solution (RS), and TBS groups. During the hemorrhagic state, their hemodynamic parameters were recorded using an Abbott i-STAT analyzer with the CG4+ cartridge (for pH, pressure of carbon dioxide, pressure of oxygen, total carbon dioxide, bicarbonate, base excess, oxygen saturation, and lactate), the CG6+ cartridge (for sodium, potassium, chloride, blood glucose, blood urea nitrogen, hematocrit, and hemoglobin), and enzyme-linked immunosorbent assay kits (calcium, magnesium, creatinine, aspartate aminotransferase, alanine aminotransferase, bilirubin, and albumin). Similar trends were found for the parameters of biochemistries, electrolytes, and blood gas, and they revealed no significant changes after blood withdrawal-induced hemorrhagic shock. However, the TBS group showed more effective ability to correct metabolic acidosis than the NS and RS groups. TBS was a feasible and safe resuscitation solution in this study and may be an alternative to NS and RS for resuscitation in hemorrhagic shock patients without liver damage.


Subject(s)
Acidosis , Alanine Transaminase , Animals , Aspartate Aminotransferases , Bilirubin , Blood Glucose , Blood Urea Nitrogen , Carbon Dioxide , Creatinine , Electrolytes , Enzyme-Linked Immunosorbent Assay , Hematocrit , Hemodynamics , Humans , Hydrogen-Ion Concentration , Liver , Magnesium , Male , Oxygen , Potassium , Rats , Resuscitation , Shock, Hemorrhagic , Sodium
12.
Article in English | WPRIM (Western Pacific) | ID: wprim-782518

ABSTRACT

Aging triggers cellular and molecular alterations, including genomic instability and organ dysfunction, which increases the risk of disease in mammals. Recently, due to the markedly growing number of aging dogs in the world, as much as 49% in total number of pet dogs, it is necessary to improve and maintain their quality of life by understanding of the biological effects of aging. Therefore, the aim of this study was to determine specific biomarkers in aging dogs as a means of defining a set of hematological/biochemical biomarkers that influence the aging process. Blood samples were collected from younger (1–3 years) and older (7–10 years) dogs of middle/large size. The hematological/biochemistry analysis was performed to evaluate parameters significantly associated with age. Enzyme-linked immunosorbent assay was used to target growth hormone (GH)/insulin growth factor-1 (IGF-1), one of the main regulators of the aging process. Declining levels of total protein and increased levels of glucose in young dogs was observed regardless of their body size. Notably, a significantly high concentration of GH and IGF-1 in the younger dogs compared to the older dogs was found in middle/large-sized dogs. GH and IGF-1 were also found at significantly high levels in large-sized dogs compared to middle-sized dogs, suggesting a similar trend to that of elderly humans. Consequently, glucose, total protein, GH, and IGF-1 were identified as potential biomarkers for regulating the aging process in large/middle-sized dogs. These findings provide an invaluable insight into the mechanism of aging for the field of aging research.


Subject(s)
Aged , Aging , Animals , Biomarkers , Body Size , Dogs , Enzyme-Linked Immunosorbent Assay , Genomic Instability , Glucose , Growth Hormone , Hematologic Tests , Humans , Insulin-Like Growth Factor I , Mammals , Quality of Life
13.
Article in English | WPRIM (Western Pacific) | ID: wprim-782294

ABSTRACT

PURPOSE: To date, many kinds of classical swine fever (CSF) vaccines have been developed to protect against this disease. However, the efficacy of these vaccines to protect the pig against field CSF strains needs to be considered, based on circulating strains of classical swine fever virus (CSFV).MATERIALS AND METHODS: Recombinant E2-CSFV protein produced by baculovirus/insect cell system was analyzed by western blots and immunoperoxidase monolayer assay. The effect of CSFV-E2 subunit vaccines was evaluated in experimental pigs with three genotypes of CSFV challenge. Anti-E2 specific and neutralizing antibodies in experimental pigs were analyzed by blocking enzyme-linked immunosorbent assay and neutralization peroxidize-linked assay.RESULTS: The data showed that CSFV VN91-E2 subunit vaccine provided clinical protection in pigs against three different genotypes of CSFV without noticeable clinical signs, symptoms, and mortality. In addition, no CSFV was isolated from the spleen of the vaccinated pigs. However, the unvaccinated pigs exhibited high clinical scores and the successful virus isolation from spleen. These results showed that the E2-specific and neutralizing antibodies induced by VN91-E2 antigen appeared at day 24 after first boost and a significant increase was observed at day 28 (p<0.01). This response reached a peak at day 35 and continued until day 63 when compared to controls. Importantly, VN91-E2 induced E2-specific and neutralizing antibodies protected experimental pigs against high virulence of CSFVs circulating in Vietnam, including genotype 1.1, 2.1, and 2.2.CONCLUSION: These findings also suggested that CSFV VN91-E2 subunit vaccine could be a promising vaccine candidate for the control and prevention of CSFV in Vietnam.


Subject(s)
Animals , Antibodies, Neutralizing , Blotting, Western , Classical Swine Fever Virus , Classical Swine Fever , Enzyme-Linked Immunosorbent Assay , Genotype , Mortality , Spleen , Swine , Vaccines , Vaccines, Subunit , Vietnam , Virulence
14.
Article in English | WPRIM (Western Pacific) | ID: wprim-782293

ABSTRACT

PURPOSE: We constructed a new canine adenovirus type 2 (CAV-2) vaccine candidate using the recently isolated Korean CAV-2 strain; we termed the vaccine APQA1701-40P and evaluated its safety and immunogenicity in dogs.MATERIALS AND METHODS: To generate the anti-CAV-2 vaccine, APQA1701 was passaged 40 times in MDCK cells growing in medium containing 5 mM urea and the virus was inactivated using 0.05% (volume per volume) formaldehyde. Two vaccines were prepared by blending inactivated APQA1701-40P with two different adjuvants; both were intramuscularly injected (twice) into guinea pigs. The safety and immunogenicity of the Cabopol-adjuvanted vaccine were evaluated in seronegative dogs. The humoral responses elicited were measured using an indirect enzyme-linked immunosorbent assay (I-ELISA), and via a virus neutralization assay (VNA).RESULTS: The new, inactivated CAV-2 vaccine strain, APQA1701-40P, lacked six amino acids of the E1b-19K protein. In guinea pigs, the Cabopol-adjuvanted vaccine afforded a slightly higher VNA titer and I-ELISA absorbance than an IMS gel-adjuvanted vaccine 4 weeks post-vaccination (p>0.05). Dogs inoculated with the former vaccine developed a significantly higher immune titer than non-vaccinated dogs.CONCLUSION: The Cabopol-adjuvanted, inactivated CAV-2 vaccine was safe and induced a high VNA titer in dogs.


Subject(s)
Adenoviruses, Canine , Amino Acids , Animals , Dogs , Enzyme-Linked Immunosorbent Assay , Formaldehyde , Guinea Pigs , Madin Darby Canine Kidney Cells , Urea , Vaccines
15.
Article in English | WPRIM (Western Pacific) | ID: wprim-782205

ABSTRACT

OBJECTIVE: Recently, the association between periodontal diseases and polycystic ovary syndrome (PCOS) has been established, and it has been revealed that visfatin levels increase in chronic periodontitis (CP) and PCOS. However, there was no study comparing the mean visfatin levels between advanced CP and PCOS. Therefore, the probable role of visfatin linking these diseases remains unknown, and this study was conducted to address this gap.METHODS: In this age- and weight-matched case-control study (cases with PCOS disease and controls without it), 110 female participants were divided into 4 groups based on clinical findings. The 1st group participants had both PCOS and advanced CP (n=30), 2nd group participants had only PCOS (n=25), 3rd group participants had only advanced CP (n=23), and 4th group comprised of healthy participants (n=32). Enzyme-linked immunosorbent assay was used to investigate visfatin levels in the gingival crevicular fluid (GCF). Data were collected and analyzed using Stata software (version 11).RESULTS: The results revealed the significant effect of both PCOS and advanced CP on visfatin levels in the GCF (P < 0.05).CONCLUSION: According to the results of this study, the visfatin level in the GCF could be the probable link of association between PCOS and advanced CP.


Subject(s)
Case-Control Studies , Chronic Periodontitis , Enzyme-Linked Immunosorbent Assay , Female , Gingival Crevicular Fluid , Healthy Volunteers , Humans , Nicotinamide Phosphoribosyltransferase , Periodontal Diseases , Polycystic Ovary Syndrome
16.
Yonsei Medical Journal ; : 73-78, 2020.
Article in English | WPRIM (Western Pacific) | ID: wprim-782121

ABSTRACT

PURPOSE: Testing for autoantibodies to extractable nuclear antigens (ENAs) plays an important role in the diagnosis and management of systemic rheumatic disease. Currently, no gold standard tests are available for detecting anti-ENAs. To address this gap, we aimed to identify an assay that exhibits satisfactory diagnostic performance in the detection of five common anti-ENAs by comparing two commonly used assays, an automated fluorescent enzyme immunoassay (FEIA) and a microplate ELISA assay.MATERIALS AND METHODS: Sera from 100 patients with systemic rheumatic disease were collected and assayed with FEIA and microplate ELISA to detect anti-ENAs. Statistical analyses were performed to check the agreement rate between the two platforms using kappa coefficients. Analytical sensitivity and specificity for each assay were calculated.RESULTS: The concordance rates between ELISA and FEIA ranged from 89% for anti-RNP to 97% for anti-Scl-70, and the kappa coefficients of the two assays were in the range of 0.44 to 0.82. Between the two assays, a significant difference in sensitivity and specificity was seen only for anti-Sm and anti-RNP, respectively.CONCLUSION: In this study, FEIA and ELISA showed comparable efficiency for detecting anti-ENAs.


Subject(s)
Antigens, Nuclear , Arthritis, Rheumatoid , Autoantibodies , Diagnosis , Enzyme-Linked Immunosorbent Assay , Fluorescence , Humans , Immunoenzyme Techniques , Lupus Erythematosus, Systemic , Rheumatic Diseases , Sensitivity and Specificity
17.
Article in English | WPRIM (Western Pacific) | ID: wprim-782080

ABSTRACT

BACKGROUND AND PURPOSE: Fibroblast growth factor 23 (FGF23) is associated with atherosclerosis via nitric-oxide-associated endothelial dysfunction and calcium-phosphate-related bone mineralization. This study aimed to determine the association of the plasma FGF23 concentration with intracranial cerebral atherosclerosis (ICAS) and extracranial cerebral atherosclerosis (ECAS).METHODS: We prospectively enrolled 262 first-ever ischemic stroke patients in whom brain magnetic resonance was performed and a blood sample acquired within 24 h after admission. Plasma FGF23 concentrations were measured using an enzyme-linked immunosorbent assay. The presence of ICAS or ECAS was defined as a ≥50% decrease in arterial diameter in magnetic resonance angiography. The burden of cerebral atherosclerosis was calculated by adding the total number of vessels defined as ICAS or ECAS.RESULTS: Our study population included 152 (58.0%) males. The mean age was 64.7 years, and the plasma FGF23 concentration was 347.5±549.6 pg/mL (mean±SD). ICAS only, ECAS only, and both ICAS and ECAS were present in 31.2% (n=82), 4.9% (n=13), and 6.8% (n=18) of the subjects, respectively. In multivariate binary and ordinal logistic analyses, after adjusting for sex, age, and variables for which p < 0.1 in the univariate analysis, the plasma FGF23 concentration (per 100 pg/mL) was positively correlated with the presence of ICAS [odds ratio (OR)=1.07, 95% CI=1.00–1.15, p=0.039], burden of ICAS (OR=1.09, 95% CI=1.04–1.15, p=0.001), and burden of ECAS (OR=1.06, 95% CI=1.00–1.12, p=0.038), but it was not significantly related to the presence of ECAS (OR=1.05, 95% CI=0.99–1.12, p=0.073).CONCLUSIONS: The plasma FGF23 may be a potential biomarker for cerebral atherosclerosis, particularly the presence and burden of ICAS in stroke patients.


Subject(s)
Atherosclerosis , Brain , Calcification, Physiologic , Enzyme-Linked Immunosorbent Assay , Fibroblast Growth Factors , Fibroblasts , Humans , Intracranial Arteriosclerosis , Magnetic Resonance Angiography , Male , Plasma , Prospective Studies , Stroke
18.
Article in English | WPRIM (Western Pacific) | ID: wprim-787244

ABSTRACT

OBJECTIVES: We aimed to evaluate the effect of insulin resistance (IR) on serum Intelectin-1 and endocrinological hormones levels in obese and non-obese women with and without polycystic ovary syndrome (PCOS) in Basrah, Iraq.METHODS: From 124 women volunteers, 60 patients with primary and 64 patients with secondary, while 56 normal ovulatory women were taken as controls. Their fasting insulin hormone, intelectin-1, anti-Mullerian hormone, luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (PRL), estradiol (E2) and testosterones (T) were determined by ELISA methods. BMI, glucose and quantitative insulin sensitivity check index as well as IR was determined by the homeostasis model assessment.RESULTS: A significant changes (P<0.05) were seen in the level of homeostasis model assessment-IR, E2 and T. Levels of anti-Mullerian hormone, LH, LH/FSH ratio and prolactin were significantly (P<0.01) increased and level of intelectin-1 and E2/T ratio were significantly (P<0.01) decreased, while quantitative insulin sensitivity check index level was not significantly different (P>0.05) between the patients (1°PCOS and 2°PCOS) and control groups. On the other hand, our data reported that FSH level was significantly (P<0.05) lower in obese and higher in non-obese patients with PCOS as compared to control group.CONCLUSION: Levels of intelectin-1 and endocrinological hormones have significantly associated with body mass index, IR and physical activity in patients and normal groups and the strategies that can modulate levels of these parameters would improve metabolic disarrangements in women with PCOS.


Subject(s)
Anti-Mullerian Hormone , Body Mass Index , Enzyme-Linked Immunosorbent Assay , Estradiol , Fasting , Female , Follicle Stimulating Hormone , Glucose , Gonadal Steroid Hormones , Hand , Homeostasis , Humans , Insulin Resistance , Insulin , Iraq , Luteinizing Hormone , Motor Activity , Polycystic Ovary Syndrome , Prolactin , Volunteers
19.
Article in English | WPRIM (Western Pacific) | ID: wprim-787140

ABSTRACT

Alzheimer's disease (AD) is the most common neurodegenerative disorder causing dementia worldwide, and is mainly characterized by aggregated β-amyloid (Aβ). Increasing evidence has shown that plant extracts have the potential to delay AD development. The plant sterol β-Sitosterol has a potential role in inhibiting the production of platelet Aβ, suggesting that it may be useful for AD prevention. In the present study, we aimed to investigate the effect and mechanism of β-Sitosterol on deficits in learning and memory in amyloid protein precursor/presenilin 1 (APP/PS1) double transgenic mice. APP/PS1 mice were treated with β-Sitosterol for four weeks, from the age of seven months. Brain Aβ metabolism was evaluated using ELISA and Western blotting. We found that β-Sitosterol treatment can improve spatial learning and recognition memory ability, and reduce plaque load in APP/PS1 mice. β-Sitosterol treatment helped reverse dendritic spine loss in APP/PS1 mice and reversed the decreased hippocampal neuron miniature excitatory postsynaptic current frequency. Our research helps to explain and support the neuroprotective effect of β-Sitosterol, which may offer a novel pharmaceutical agent for the treatment of AD. Taken together, these findings suggest that β-Sitosterol ameliorates memory and learning impairment in APP/PS1 mice and possibly decreases Aβ deposition.


Subject(s)
Alzheimer Disease , Amyloid , Animals , Blood Platelets , Blotting, Western , Brain , Cognition Disorders , Dementia , Dendritic Spines , Enzyme-Linked Immunosorbent Assay , Excitatory Postsynaptic Potentials , Learning , Memory , Metabolism , Mice , Mice, Transgenic , Neurodegenerative Diseases , Neurons , Neuroprotective Agents , Plant Extracts , Plants , Plaque, Amyloid , Spatial Learning
20.
Article in English | WPRIM (Western Pacific) | ID: wprim-811069

ABSTRACT

PURPOSE: Different characteristics of airway microbiome in asthmatics may lead to differential immune responses, which in turn cause eosinophilic or neutrophilic airway inflammation. However, the relationships among these factors have yet to be fully elucidated.METHODS: Microbes in induced sputum samples were subjected to sequence analysis of 16S rRNA. Airway inflammatory phenotypes were defined as neutrophils (>60%) and eosinophils (>3%), and inflammation endotypes were defined by levels of T helper (Th) 1 (interferon-γ), Th2 (interleukin [IL]-5 and IL-13), Th-17 (IL-17), and innate Th2 (IL-25, IL-33, and thymic stromal lymphopoietin) cytokines, inflammasomes (IL-1β), epithelial activation markers (granulocyte-macrophage colony-stimulating factor and IL-8), and Inflammation (IL-6 and tumor necrosis factor-α) cytokines in sputum supernatants was assessed by enzyme-linked immunosorbent assay.RESULTS: The numbers of operational taxonomic units were significantly higher in the mixed (n = 21) and neutrophilic (n = 23) inflammation groups than in the paucigranulocytic inflammation group (n = 19; p < 0.05). At the species level, Granulicatella adiacens, Streptococcus parasanguinis, Streptococcus pneumoniae, Veillonella rogosae, Haemophilus parainfluenzae, and Neisseria perflava levels were significantly higher in the eosinophilic inflammation group (n = 20), whereas JYGU_s levels were significantly higher in the neutrophilic inflammation group compared to the other subtypes (P < 0.05). Additionally, IL-5 and IL-13 concentrations were correlated with the percentage of eosinophils (P < 0.05) and IL-13 levels were positively correlated with the read counts of Porphyromonas pasteri and V. rogosae (P < 0.05). IL-1β concentrations were correlated with the percentage of neutrophils (P < 0.05). had a tendency to be positively correlated with the read count of JYGU_s (P = 0.095), and was negatively correlated with that of S. pneumoniae (P < 0.05).CONCLUSIONS: Difference of microbial patterns in airways may induce distinctive endotypes of asthma, which is responsible for the neutrophilic or eosinophilic inflammation in asthma.


Subject(s)
Asthma , Colony-Stimulating Factors , Cytokines , Enzyme-Linked Immunosorbent Assay , Eosinophils , Haemophilus parainfluenzae , Inflammasomes , Inflammation , Interleukin-13 , Interleukin-33 , Interleukin-5 , Microbiota , Necrosis , Neisseria , Neutrophils , Phenotype , Pneumonia , Porphyromonas , Sequence Analysis , Sputum , Streptococcus , Streptococcus pneumoniae , Veillonella
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