Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 150.925
Filtrar
1.
Medicine (Baltimore) ; 99(1): e18462, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31895778

RESUMO

Proinflammatory interleukin-26 (IL-26) is involved in chronic inflammation; however, the role of IL-26 in chronic hepatitis B (CHB) remains unknown.In this study, serum IL-26 was quantified in a cohort of CHB patients at baseline and during telbivudine (LdT) treatment.Our results showed that the serum IL-26 level was significantly elevated in CHB patients compared with that in healthy controls and was time-dependently decreased during LdT treatment, accompanying hepatitis B e antigen (HBeAg) seroconversion and reduced serum levels of hepatitis B virus (HBV) DNA, aspartate transaminase, and alanine transaminase across baseline and treatment. In addition, the serum level of IL-26 exhibited a similar declining trend to that of T helper 17 (Th17) cell-secreted IL-17 during LdT treatment in CHB patients. The percentage of IL-26-expressing CD4 cells was significantly higher than that of IL-26-expressing CD4 cells isolated from the peripheral blood mononuclear cells of CHB patients, suggesting that serum IL-26 might be mainly released from CD4 T cells. Furthermore, the baseline mRNA levels of IL-26 and orphan nuclear receptor RORγt-an important transcription factor expressed by Th17 cells-were positively correlated and displayed the same declining trend across the baseline and LdT treatment in CHB patients, suggesting that Th17 cells could be a possible cellular source of the increased serum IL-26 in CHB patients.Taken together, our results suggest that serum IL-26, possibly produced by Th17 CD4 cells, is a novel and potential biomarker for CHB prognosis and treatment.


Assuntos
Vírus da Hepatite B/imunologia , Hepatite B Crônica/sangue , Hepatite B Crônica/imunologia , Interleucinas/sangue , Adulto , Antivirais/uso terapêutico , Linfócitos T CD4-Positivos/imunologia , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Feminino , Hepatite B Crônica/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Telbivudina/uso terapêutico , Células Th17/imunologia , Adulto Jovem
2.
Vet Parasitol ; 277: 109012, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31884316

RESUMO

Early in vivo diagnosis of bovine besnoitiosis is crucial for the success of control programmes. However, diagnosis in acutely infected animals is hindered by the low sensitivity of the available serological tools. In this study, a novel ELISA to detect specific anti-Besnoitia besnoiti IgM antibodies was developed. The usefulness of this tool together with an avidity ELISA were studied with a well-coded sera panel from experimentally and naturally infected cattle. First, the kinetics of specific IgM levels were determined in experimentally infected calves during the acute and chronic infection. Next, IgM levels were determined in naturally infected cattle with either acute or chronic infection. Finally, the IgG avidity index was monitored in both experimentally and naturally infected cattle. Specific IgM antibodies were detected prior to specific IgG antibodies (7-19 days vs. 17-26 days post-infection). A prompt IgM response was associated with the end of the febrile stage in experimentally infected calves. Naturally and experimentally infected animals with acute clinical signs tested IgM-positive but IgG-negative, followed by IgG seroconversion 2-3 weeks later. Chronically infected cattle developed both IgM and IgG specific antibodies. Moreover, a progressive increase in the avidity index (AI) was observed in all experimentally infected calves during the course of the experimental trials. However, a low AI coincided with visible tissue cysts. Low avidity values were also detected when naturally infected cattle with acute clinical signs seroconverted, in contrast to a high AI detected in chronically infected cattle. In summary, IgM and avidity ELISAs improved the early in vivo diagnosis of bovine besnoitiosis. IgM-positive but IgG-negative results were indicative of an acute infection, whereas IgG positive results accompanied by low avidity values confirmed a recent infection.


Assuntos
Doenças dos Bovinos/diagnóstico , Coccidiose/veterinária , Doença Aguda , Animais , Anticorpos Antiprotozoários/sangue , Bovinos , Coccidiose/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Imunoglobulina M/sangue
3.
Food Chem ; 307: 125565, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31630022

RESUMO

The effectiveness of some non-specific proteases in reducing raw peanut allergenicity was investigated. Peanut kernels were treated by Alcalase, papain, Neutrase and bromelain, respectively. The residues of major peanut allergens Ara h 1, Ara h 2 and Ara h 6 were determined by sandwich ELISA and SDS-PAGE, and the allergenicities of treated peanuts were compared to that of untreated peanuts by western blot. All tested proteases were effective in reducing Ara h 1, but their effectiveness in hydrolyzing Ara h 2 and Ara h 6 varied greatly. The maximal reductions of extractable Ara h 1, Ara h 2 and Ara h 6 were 100%, 100% and 99.8%, respectively, achieved by Alcalase hydrolysis. Alcalase was more effective in overall allergenicity reduction; bromelain and Neutrase were the least effective in reducing Ara h 2 and Ara h 6, respectively. The hydrolysis of original allergens also produced some smaller peptides with strong IgE-binding.


Assuntos
Alérgenos/metabolismo , Arachis/química , Imunoglobulina E/imunologia , Hipersensibilidade a Amendoim/prevenção & controle , Peptídeo Hidrolases/metabolismo , Albuminas 2S de Plantas/análise , Albuminas 2S de Plantas/imunologia , Albuminas 2S de Plantas/metabolismo , Alérgenos/análise , Alérgenos/imunologia , Antígenos de Plantas/análise , Antígenos de Plantas/imunologia , Antígenos de Plantas/metabolismo , Western Blotting , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Humanos , Proteínas de Membrana/análise , Proteínas de Membrana/imunologia , Proteínas de Membrana/metabolismo , Proteínas de Plantas/análise , Proteínas de Plantas/imunologia , Proteínas de Plantas/metabolismo
4.
Biosci Biotechnol Biochem ; 84(1): 1-16, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31538538

RESUMO

Recent investigations suggest that soluble oligomeric amyloid ß (Aß) species may be involved in early onset of Alzheimer's disease (AD). Using systematic proline replacement, solid-state NMR, and ESR, we identified a toxic turn at position 22 and 23 of Aß42, the most potent neurotoxic Aß species. Through radicalization, the toxic turn can induce formation of the C-terminal hydrophobic core to obtain putative Aß42 dimers and trimers. Synthesized dimer and trimer models showed that the C-terminal hydrophobic core plays a critical role in the formation of high molecular weight oligomers with neurotoxicity. Accordingly, an anti-toxic turn antibody (24B3) that selectively recognizes a toxic dimer model of E22P-Aß42 was developed. Sandwich enzyme-linked immunosorbent assay with 24B3 and 82E1 detected a significantly higher ratio of Aß42 with a toxic turn to total Aß42 in cerebrospinal fluid of AD patients compared with controls, suggesting that 24B3 could be useful for early onset of AD diagnosis.


Assuntos
Doença de Alzheimer/diagnóstico , Doença de Alzheimer/tratamento farmacológico , Peptídeos beta-Amiloides/líquido cefalorraquidiano , Peptídeos beta-Amiloides/imunologia , Anticorpos Monoclonais/uso terapêutico , Fragmentos de Peptídeos/líquido cefalorraquidiano , Fragmentos de Peptídeos/imunologia , Doença de Alzheimer/líquido cefalorraquidiano , Peptídeos beta-Amiloides/química , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Humanos , Camundongos , Modelos Moleculares , Emaranhados Neurofibrilares/química , Fragmentos de Peptídeos/química , Placa Amiloide/química , Prolina/química , Agregados Proteicos , Agregação Patológica de Proteínas , Estrutura Terciária de Proteína
5.
Bioelectrochemistry ; 131: 107397, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31706117

RESUMO

A new polyclonal antibody that recognizes the CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4-EPSPS), which provides resistance to glyphosate in soybean (Roundup Ready®, RR soybean), was produced. New Zealand rabbits were injected with a synthetic peptide (Pc_312-324, (PEP)) present in the soybean CP4-EPSPS protein. The anti-PEP antibodies production was evaluated by electrophoresis (SDS-PAGE) and an enzyme-linked immunosorbent assay (ELISA) was developed in order to study their specificity. The ELISA showed that the polyclonal antibody was specific to PEP. In addition, the anti- PEP was immobilized onto a gold disk electrode and the antigen-antibody interaction was evaluated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Moreover, the EIS showed that the electron transfer resistance of the modified electrode increased after incubation with solutions containing CP4-EPSPS protein from RR transgenic soybean, while no changes were detected after incubation with no-RR soybean proteins. These results suggest that the CP4-EPSPS was immobilized onto the electrode, due to the specific interaction with the anti-PEP. These results show that this antigen-antibody interaction can be detected by electrochemical techniques, suggesting that the anti-PEP produced can be used in electrochemical immunosensors development to quantify transgenic soybean.


Assuntos
Anticorpos/metabolismo , Formação de Anticorpos , Técnicas Eletroquímicas/métodos , Peptídeos/metabolismo , Plantas Geneticamente Modificadas/imunologia , Soja/imunologia , Técnicas Biossensoriais , Ensaio de Imunoadsorção Enzimática/métodos
6.
Einstein (Sao Paulo) ; 18: eAO4784, 2020.
Artigo em Inglês, Português | MEDLINE | ID: mdl-31553356

RESUMO

OBJECTIVE: To evaluate the effect of three types of muscular resistance training on adiposity, inflammation levels and insulin activity in Swiss mice with fat-rich diet-induced obesity. METHODS: Lean and obese male Swiss mice were selected and allocated to one of eight groups comprising eight mice each, as follows: standard diet + no training; standard diet + muscular resistance training; standard diet + hypertrophy training; standard diet + strength training; high-fat diet + no training; high-fat diet + muscular resistance training; high-fat diet + hypertrophy training; high-fat diet + strength training. The training protocol consisted of stair climbing for a 10-week period. Blood samples were collected for lactate analysis, glucose level measurement and insulin tolerance test. After euthanasia, adipose tissues were removed and weighed for adiposity index determination. Fragments of epididymal adipose tissue were then embedded for histological analysis or homogenized for tumor necrosis factor alpha level determination using the ELISA method. RESULTS: Ausency of differences in total training volume and blood lactate levels overall emphasize the similarity between the different resistance training protocols. Body weight loss, reduced adipocyte area and lower adiposity index were observed in trained obese mice, regardless of training modality. Different training protocols also improved insulin sensitivity and reduced inflammation levels. CONCLUSION: Resistance training protocols were equally effective in reducing body fat, inflammation levels and insulin resistance in obese mice.


Assuntos
Adiposidade/fisiologia , Hipertrofia/fisiopatologia , Inflamação/fisiopatologia , Resistência à Insulina/fisiologia , Exercícios de Alongamento Muscular/métodos , Obesidade/fisiopatologia , Condicionamento Físico Animal/fisiologia , Tecido Adiposo Branco/fisiopatologia , Animais , Glicemia/análise , Peso Corporal/fisiologia , Dieta Hiperlipídica , Ensaio de Imunoadsorção Enzimática , Masculino , Camundongos , Camundongos Obesos , Reprodutibilidade dos Testes , Treinamento de Resistência/métodos , Fatores de Tempo , Fator de Necrose Tumoral alfa/análise
7.
Medicine (Baltimore) ; 98(50): e17814, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31852062

RESUMO

The purpose of our research was to evaluate diagnostic performance of serum microRNA-135a (miR-135a) in non-small cell lung cancer (NSCLC).Quantitative real time-polymerase chain reaction was employed to detect the expression serum of miR-135a in NSCLC patients and controls. The influence of serum miR-135a level on clinical characteristics of NSCLC patients was explored through the Chi-square test. Serum carcinoembryonic antigen (CEA) level was estimated via enzyme-linked immunosorbent assay. Receiver operating characteristic (ROC) curve was plotted to elucidate diagnostic roles of serum miR-135a and CEA in NSCLC.The expression level of serum miR-135a was significantly lower in NSCLC patients than in healthy controls (0.40 ±â€Š0.29 vs 1.00 ±â€Š0.40, P < .001). Moreover, miR-135a expression was related to lymph node metastasis (P = .021), tumor differentiation (P = .020), and tumor node metastasis stage (P = .031). ROC curve showed serum miR-135a level could discriminate NSCLC patients from healthy controls (P < .0001) with a corresponding cutoff value of 0.665, and a sensitivity and specificity of 81.3% and 83.1%, respectively. The area under the curve was 0.888. In diagnosis analysis on the combination of miR-135a and CEA, when its specificity was maintained at 90%, diagnosis cut-off point reached 0.678.Serum miR-135a level is significantly downregulated in NSCLC and serves as a potential diagnostic biomarker for the disease.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Neoplasias Pulmonares/diagnóstico , MicroRNAs/sangue , Estadiamento de Neoplasias/métodos , Idoso , Biomarcadores Tumorais/sangue , Carcinoma Pulmonar de Células não Pequenas/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Neoplasias Pulmonares/sangue , Masculino , Pessoa de Meia-Idade , RNA Neoplásico/genética , Curva ROC , Reação em Cadeia da Polimerase em Tempo Real
8.
Beijing Da Xue Xue Bao Yi Xue Ban ; 51(6): 1019-1024, 2019 Dec 18.
Artigo em Chinês | MEDLINE | ID: mdl-31848497

RESUMO

OBJECTIVE: Antibodies against carbamylated protein (anti-CarP) were found to be a promising marker to evaluate joint damage and disease activity in patients with rheumatoid arthritis (RA). However, whether anti-CarP antibodies were present in systemic lupus erythematosus (SLE) remained ambiguity. We have therefore undertaken this study to assess the levels of serum anti-CarP antibodies and to evaluate their clinical value in SLE. METHODS: Serum levels of antibodies against carbamylatedfibrinogen (anti-CarP) were measured by enzyme-linked immunosorbent assay (ELISA) in 105 SLE patients and 73 healthy controls. Other clinical and laboratory measurements of the SLE patients were collected from medical records. Data analyses between anti-CarP antibodies and other laboratory measurements were performed using SPSS software for Windows 24.0. RESULTS: The levels of serum anti-CarP antibodies in the patients with SLE were significantly higher than those in the healthy controls (P<0.05). There were significant differences between the anti-CarP-positive group and anti-CarP-negative group in many clinical features. The disease duration, values of ESR, CRP, RF, anti-cardiolipin, anti-dsDNA, D-dipolymer, IgA and IgG were significantly higher in the anti-CarP-positive group compared with the negative group (P<0.05). Conversely, the values of complement 3, complement 4, peripheral blood RBC, and hemoglobin were significantly lower in anti-CarP-positive group than in the negative group(P<0.05). Moreover, the incidence of increase of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF), D-dipolymer, decrease of peripheral blood RBC, hemoglobin, complement 3, complement 4, and positive rate of anti-dsDNA were significant different between the two groups(P<0.05). The positive rate of anti-CarP (21.9%) was higher than that of anti-Sm (15.24%), and close to anti-ribosomal P protein (22.86%) in our SLE patients. In addition, anti-CarP antibody was present in the SLE patients lacking the disease specific antibodies, including anti-Sm (anti-CarP positive rate 20.2%, 18/89), anti-dsDNA (anti-CarP positive rate 9.3%, 4/43), anti-nucleosome (anti-CarP positive rate 12.5%, 6/48), and anti-ribosomal P protein antibody (anti-CarP positive rate 20.9%, 17/81). Moreover, the high levels of anti-CarP antibodies were correlated with short disease duration, low C3, C4, RBC, and hemoglobin (P<0.05), high ESR, CRP, IgA, IgG, RF, anti-cardiolipin, anti-dsDNA, and D-dipolymer (P<0.05). CONCLUSION: The level of anti-CarP antibody was increased in the serum of patients with SLE. There were correlations between anti-CarP antibodies and clinical and laboratory indicators of SLE patients.


Assuntos
Lúpus Eritematoso Sistêmico , Autoanticorpos , Sedimentação Sanguínea , Ensaio de Imunoadsorção Enzimática , Fibrinogênio , Humanos , Fator Reumatoide
9.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 31(5): 510-512, 2019 Aug 16.
Artigo em Chinês | MEDLINE | ID: mdl-31713380

RESUMO

OBJECTIVE: To examine the effect of low temperature on trehalose and trehalase levels in Culex pipiens pallens. METHODS: The fourth instar larvae and female adult mosquitoes of Cx. pipiens pallens were exposed at 4 ℃ for 0, 1, 3, 6, 12, 18, 24 h and 0, 1, 3, 6, 12, 18, 24, 36, 48, 72 h, respectively. Then, the trehalose and trehalase contents were detected by enzyme-linked immunosorbent assay (ELISA) in mosquitoes. RESULTS: The contents of trehalose and trehalase significantly increased in the larval and female adult mosquitoes post-exposure to low temperature. The changing trend of trehalose levels was consistent in the larval and female adult mosquitoes, and the highest levels were (2.458 8 ± 0.379 2) mg/g and (2.825 7 ± 0.211 1) mg/g 3 h post-exposure to low temperature, respectively. The trehalose and trehalase levels fluctuated greatly within the first 6 h post-exposure to low temperature. Following adaptation for a period of time, the trehalose and trehalase levels remained at a relatively high level. CONCLUSIONS: Low temperature may induce the production of trehalose and trehalase in Cx. pipiens pallens, and the trehalose and trehalase may play an important role in the improvement of the cold resistance.


Assuntos
Culex , Temperatura Ambiente , Trealase , Trealose , Animais , Culex/enzimologia , Culex/genética , Ensaio de Imunoadsorção Enzimática , Feminino , Regulação Enzimológica da Expressão Gênica/fisiologia , Larva , Trealase/metabolismo , Trealose/metabolismo
10.
Zhonghua Jie He He Hu Xi Za Zhi ; 42(11): 845-851, 2019 Nov 12.
Artigo em Chinês | MEDLINE | ID: mdl-31694095

RESUMO

Objective: To explore the role of S100A8, the receptor for advanced glycation endproducts (RAGE) and Caveolin-1 in neutrophilic asthmatic rats, and to further study the intervention of roxithromycin and the possible mechanisms. Methods: Male Brown Norway rats were randomly assigned to a control group, an asthma group and a Roxithromycin group. The asthmatic rat model was established by intraperitoneal injection of ovalbumin (OVA) and Freund's complete adjuvant (FCA) mixture, and aerosol inhalation of OVA. Rats in the Roxithromycin group were given roxithromycin injection 30 mg/kg 30 minutes before each challenge. Rats in the control and the asthma groups were replaced with equal volumes of saline, respectively. Bronchoalveolar lavage fluid (BALF) neutrophil percentage (Neu%) and pathological changes of pulmonary tissue (hematoxylin-eosin, HE staining) were measured to confirm the establishment of asthmatic models. The concentration of inflammatory cytokines and S100A8 were quantified by enzyme-linked immunosorbent assay (ELISA), and the expression of Caveolin-1 and RAGE at protein levels were detected by immunohistochemistry and Western blot. Results: Neu% in BALF of the asthma group was significantly higher than those of the control group, and Neu% in the Roxithromycin group was lower than the asthma group (all P<0.01). Pulmonary histology revealed that there were a large number of inflammatory cells infiltrated in the bronchial and perivascular, pulmonary interstitial and alveolar spaces, and the bronchial wall and smooth muscles were thickened obviously in the asthma group. Rats in the Roxithromycin group showed milder inflammation and airway remodeling change than the asthma group. There was no obvious pathological damage in the control group. The concentration of IL-6 and IL-17 in BALF and serum of rats in the asthma group were significantly higher than those in the control group (P<0.01), and Roxithromycin inhibited the high expression of these cytokines (P<0.05). The expression of S100A8 and RAGE in the asthma group were significantly higher than those in the control group [(20.6±4.4) vs (7.1±2.0) ng/L; (885±118) vs (462±102) ng/L; (14.2±1.7) vs (7.6±1.8) ng/L; (774±166) vs (406±69) ng/L, all P<0.05], and Roxithromycin inhibited the high expression of these proteins [(14.3±3.7) vs (20.6±4.4) ng/L; (650±53) vs (885±118) ng/L; (10.4±1.2) vs (14.2±1.7) ng/L; (560±64) vs (728±72) ng/L] (all P<0.05). Meanwhile, the expression of Caveolin-1 in the asthma group was significantly lower than that in the control group (P<0.01), and Roxithromycin up-regulated its expression (P<0.01). Correlation analysis showed that there was a significantly positive correlation between the expression of S100A8 and RAGE (r=0.706, P<0.01), while there was a significantly negative correlation between the expression of S100A8 and Caveolin-1 (r=-0.775, P<0.01), and between the expression of Caveolin-1 and RAGE (r=-0.919, P<0.01). Conclusion: S100A8 and Caveolin-1 may play an important role in neutrophilic asthma via RAGE, and Roxithromycin may exerts anti-inflammatory effects and inhibition of airway remodeling partly through this signaling pathway.


Assuntos
Antibacterianos/farmacologia , Asma/tratamento farmacológico , Calgranulina A/efeitos dos fármacos , Caveolina 1/efeitos dos fármacos , Roxitromicina/farmacologia , Remodelação das Vias Aéreas , Animais , Antibacterianos/administração & dosagem , Western Blotting , Líquido da Lavagem Broncoalveolar , Calgranulina A/metabolismo , Caveolina 1/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Imuno-Histoquímica , Pulmão/fisiopatologia , Masculino , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Ovalbumina , Ratos , Receptor para Produtos Finais de Glicação Avançada , Roxitromicina/administração & dosagem
11.
Anticancer Res ; 39(11): 6317-6324, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31704862

RESUMO

BACKGROUND/AIM: The aim of this study was to evaluate N-acetylgalactosamine-6-sulfatase (GALNS) as a new biomarker candidate for detecting lung cancer. Glycodelin or PAEP, the serum levels of which are known to be elevated in lung and other cancers, served as a benchmark for comparison. PATIENTS AND METHODS: A total of 170 serum samples from healthy controls and patients with pneumonia, lung cancer, breast cancer, colon cancer, liver cancer, and head and neck cancer were analyzed for the levels of GALNS and PAEP by ELISA. RESULTS: The median serum levels of GALNS and PAEP in all cancer types as well as pneumonia patients were significantly higher than those of the healthy controls. CONCLUSION: In addition to previously known cancers, the median serum levels of PAEP were also found to be higher in liver and head and neck cancer patients. GALNS and PAEP are promising general biomarkers for multiple cancers and deserve further evaluation.


Assuntos
Biomarcadores Tumorais/sangue , Condroitina Sulfatases/sangue , Glicodelina/sangue , Neoplasias Pulmonares/sangue , Área Sob a Curva , Benchmarking , Neoplasias da Mama/sangue , Estudos de Casos e Controles , Linhagem Celular Tumoral , Neoplasias do Colo/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Neoplasias de Cabeça e Pescoço/sangue , Humanos , Neoplasias Hepáticas/sangue , Pulmão/metabolismo , Neoplasias Pulmonares/diagnóstico , Masculino , Pneumonia/sangue
12.
Klin Lab Diagn ; 64(11): 654-658, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31747492

RESUMO

The developing and the testing results of an immunochromatographic test for the D-dimer qualitative determination were presents in the article. The test was approved blood plasma samples in comparison with a quantitative enzyme-linked immunosorbent assay (ELISA). The results of assay with developed test were the same with ELISA results for 87,1% and 100% for samples with increased (more than 400 ng/ml FEU) and normal concentration of D-dimer, respectively. The immunochromatographic test for determination of D-dimer can be included in the diagnostic strategy as a cut test after the assessment of venous thromboembolism risk.


Assuntos
Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Imunoensaio , Tromboembolia Venosa/diagnóstico , Ensaio de Imunoadsorção Enzimática , Humanos , Sensibilidade e Especificidade
13.
Klin Lab Diagn ; 64(11): 686-689, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31747499

RESUMO

Based on experimental studies with 10-fold dilutions of two the strains TBEV of the Far Eastern subtype, different in moleculargenetic characteristic complex data of simultaneously taking into account three indicators of their verification (virus titer, ELISA and PCR-RT) were obtained. The efficiency of detecting a genetic marker in PCR compared with ELISA for a weakly pathogenic strain with defects in the genetic structure was higher by a factor of 10, and for a highly pathogenic strain by a factor of 5,000. At the same time, positive results in both reactions with respect to two strains were detected with a virus titer of not less than 1-1.5 log TCID50, i.e. this level of virus in the sample is defined as epidemically significant. An algorithm for conducting research on the verification of TBEV is proposed: 1) Ticks collected from vegetation can be examined by ELISA or by PCR. All positive results can be summarized and considered viral ticks; 2) All samples with positive results only in PCR or ELISA must be investigated in two reactions in order to obtain confirmation of the possible infectivity of the pathogen; 3) To obtain a fast complex result of infection of the removed ticks from patients or blood after a tick bite, studies should be carried out simultaneously in two reactions simultaneously in ELISA and PCR; 4) Isolation virus should be carried out in biological samples (ticks collected from vegetation, ticks removed from patients, the blood of patients with suspected TBE, mammals) only with the same results in PCR and ELISA. Thus, such an approach to verifying TBEV in a tick or in the blood of patients will improve the reliability of laboratory diagnostics, identifying not only markers of TBEV, but also determining the infectivity of the pathogen, which may be the basis for the appointment of early intensive antiviral therapy.


Assuntos
Vírus da Encefalite Transmitidos por Carrapatos/genética , Encefalite Transmitida por Carrapatos/diagnóstico , Ixodes/virologia , Animais , Vírus da Encefalite Transmitidos por Carrapatos/patogenicidade , Ensaio de Imunoadsorção Enzimática , Humanos , RNA Viral , Reprodutibilidade dos Testes
14.
Arch Virol ; 164(12): 3007-3017, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31598846

RESUMO

Classical swine fever (CSF) is an important viral disease of domestic pigs and wild boar. The structural proteins E2 and Erns of classical swine fever virus (CSFV), which participate in the attachment of the virion to the host cell surface and its subsequent entry, are immunogenic. The E2 and Erns proteins are used for diagnosis and the development of vaccines against CSFV infection in swine. Newcastle disease virus (NDV) has been successfully used as a viral vector to express heterologous proteins. In the present study, the E2 and Erns proteins of CSFV were expressed in cell culture as well as embryonated chicken eggs, using recombinant NDV (rNDV). Rescued rNDV expressing the E2 and Erns proteins induced the production of CSFV-neutralizing antibodies upon intranasal vaccination of pigs. Serum samples from vaccinated animals were found to neutralize both homologous and heterologous CSFV strains. Furthermore, rNDV expressing the E2 and Erns proteins of CSFV was used to develop an indirect ELISA, which was used to measure the the antibody titers of randomly collected serum samples. The results suggested that the ELISA based on rNDV-expressed E2 and Erns proteins could be used to screen for CSFV infections. This study shows that rNDV-based expression of CSFV antigens is potentially applicable for development of vaccines and diagnostic tests for CSFV infection. This approach could be an economically favorable alternative to the existing vaccine and diagnostics for CSFV in pigs.


Assuntos
Vírus da Febre Suína Clássica/imunologia , Peste Suína Clássica/diagnóstico , Testes Diagnósticos de Rotina/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Glicoproteínas de Membrana/sangue , Vírus da Doença de Newcastle/genética , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Peste Suína Clássica/sangue , Peste Suína Clássica/imunologia , Peste Suína Clássica/virologia , Vírus da Febre Suína Clássica/genética , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Vírus da Doença de Newcastle/imunologia , Recombinação Genética , Suínos
15.
Pan Afr Med J ; 33: 163, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31565125

RESUMO

Viral hepatitis is a serious public health problem. Its epidemiology is not precisely known in Morocco. Our objective was to assess the prevalence of HBV and HCV in a particular population of "blood donors" at the Regional Blood Transfusion Centre in Oujda. A retrospective study was conducted from May 1, 2013 to May 31, 2015. Thirty-one thousand nine hundred and fifty-two blood donors were tested. Antigen detection was made according to ELISA technique (MonolisaTMHBs Ag ULTRA). HCV research was performed by ELISA using the kit « Monolisa HCV Ag-Ab ULTRA ¼. 177 blood donors included, they are divided into 155 male (87.6%) and 22 female (12.4%) subjects with a ratio of 7. The average age was 37.64 ± 12 years. Six cases were positive for HCV with an overall prevalence of 0.02%. The population study by sex shows a prevalence of 0.004% for 23177 male sera and 0.057% for 8775 female sera. Six donors were HCV positive, of which 05 were female (83.33%) and one was male (16.66%). The average age was 43 ± 14 years. Co-infection with HCV HBV-HCV and HCV-Syphilis and HCV-HIV are absent. Co-infection with HBV and HIV was found in one case. HBV-syphilis co-infection was found in 04 cases. Chronic viral hepatitis is a real global health problem. Its prevalence is currently estimated at 0.55% for HBV and 0.02% for HCV, reclassifying Morocco as a low endemic area. The prevention remains the most effective method to successfully control HBV and HCV infection.


Assuntos
Doadores de Sangue/estatística & dados numéricos , Infecções por HIV/epidemiologia , Hepatite B/epidemiologia , Hepatite C/epidemiologia , Adolescente , Adulto , Coinfecção , Ensaio de Imunoadsorção Enzimática , Feminino , Antígenos de Superfície da Hepatite B/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Marrocos/epidemiologia , Prevalência , Estudos Retrospectivos , Sífilis/epidemiologia , Adulto Jovem
16.
Medicine (Baltimore) ; 98(38): e16991, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31567936

RESUMO

BACKGROUND: Interleukin 12 (IL-12) and interleukin 12 receptor (IL12R), key inflammatory cytokines in the immune system, participate in bridging the innate immunity and adaptive immunity. No previous work has reported the role of IL-12 and IL12R in high-risk human papillomavirus (hrHPV) susceptibility. The purpose of this study was to investigate the association of IL-12, IL12R polymorphisms, and serum IL-12 levels with hrHPV susceptibility in rural women from Luohe, Henan, China. METHODS: Two hundred sixty cases with hrHPV infection and 260 healthy controls were selected. Enzyme-linked immunosorbent assays were used to detect the serum IL-12 levels, and the polymorphisms of IL12B rs3212227, IL12RB1 rs393548, and IL12RB1 rs436857 were determined using DNA sequencing. RESULTS: The serum IL-12 levels were significantly lower in cases with hrHPV infection compared with those in healthy controls (P < .01).There was no significant difference in IL12 rs3212227, IL12RB1rs436857, and IL12RB1rs393548 genotype and allele frequencies between cases and controls (P > .05). Furthermore, with respect to the IL12 rs3212227 polymorphism with serum IL-12 levels, although serum IL-12 levels were lower in cases than in controls, we did not find any differences between serum IL-12 levels and genotypes in cases(P > .05). CONCLUSIONS: Our data demonstrates that low serum IL-12 levels may be associated with hrHPV susceptibility but are not associated with IL-12 gene polymorphisms; furthermore, IL-12 and IL12R gene polymorphisms may not contribute susceptibility to hrHPV in rural women from Luohe, Henan, China.


Assuntos
Predisposição Genética para Doença , Subunidade p40 da Interleucina-12/genética , Infecções por Papillomavirus/genética , Receptores de Interleucina-12/genética , Adulto , Idoso , Grupo com Ancestrais do Continente Asiático , Estudos de Casos e Controles , China , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Subunidade p40 da Interleucina-12/sangue , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Receptores de Interleucina-12/sangue , População Rural
17.
Medicine (Baltimore) ; 98(38): e17146, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31567954

RESUMO

Chronic kidney disease (CKD) will progress to end stage without treatment, the decline off renal function may not linear. A sensitive marker such as soluble urokinase-type plasminogen activator receptors (suPARs) may allow potential intervention and treatment in earlier stages of CKD. OBJECTIVES: This study was designed to measure plasma (suPAR) in patients with CKD with different stages and to find its correlation with the disease severity. METHODS: This study was conducted on 114 subjects, 84 were patients with different stages and different causes of CKD, and 30 healthy subjects as controls. Blood urea, serum creatinine, serum high-sensitive C-reactive protein, estimated glomerular filtration rate, and 24 hours proteinuria were measured, renal biopsy was done for all patients, and plasma (suPAR) was measured using enzyme-linked immunosorbent assay. RESULTS: suPAR plasma levels were significantly higher in patients with CKD (7.9 ±â€Š3.82 ng/mL) than controls (1.76 ±â€Š0.77 ng/mL, P < .001). suPAR correlated with the disease severity. In stage 1 to 2 group, it was 3.7 ±â€Š1.5 ng/mL, in stage 3 to 4, it was 10.10 ±â€Š1.22 ng/mL, and in stage 5 group, it was 12.34 ±â€Š0.88 ng/mL; the difference between the 3 groups was highly significant (P < .001). A cutoff point 2.5 ng/mL of suPAR was found between controls and stage 1 group. According to the cause of CKD, although patients with obstructive cause and those with focal glomerulosclerosis had the higher levels 9.11 ±â€Š3.32 ng/mL and 8.73 ±â€Š3.19 ng/mL, respectively, but there was no significant difference between patients with CKD according to the cause of the CKD. CONCLUSION: Plasma (suPAR) increased in patients with CKD and correlated with disease severity.


Assuntos
Receptores de Ativador de Plasminogênio Tipo Uroquinase/sangue , Insuficiência Renal Crônica/sangue , Adulto , Idoso , Proteína C-Reativa/análise , Estudos de Casos e Controles , Creatinina/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Taxa de Filtração Glomerular , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência Renal Crônica/patologia , Índice de Gravidade de Doença , Ureia/sangue , Adulto Jovem
18.
Klin Lab Diagn ; 64(9): 530-535, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31610104

RESUMO

HHV-7 (Human Herpes virus type 7) is a relatively recently discovered, ubiquitous beta herpes virus. In Russia, the diagnosis of HHV-7 infection is carried out by PCR, which determines the DNA in serum or plasma, if there are clinical indications. Laboratory serological tests for HHV-7 are not performed. Currently, in the scientific literature, information on clinical studies of HHV-7 infection, its epidemiology, and its role in the etiopathogenesis of various diseases is insufficient. This article presents the results of the development of a test kit "ELISA-HHV-7-IgG", designed to detect specific IgG to HHV-7 by enzyme-linked immunosorbent assay (ELISA). Studies of the sensitivity and specificity of the developed test kit, including potential cross-reactive samples positive for other herpesvirus infections, were carried out. In the context of serological diagnostics, we studied samples of the sera of children (n = 167) and old people (n = 238) for the presence of IgG to HHV-7 with confirmation of the results in the indirect immunofluorescence reaction and the immune blotting reaction. The research results are evidence of the possibility of using the developed test kit for serological diagnosis of HHV-7 infection.


Assuntos
Ensaio de Imunoadsorção Enzimática , Herpesvirus Humano 7 , Infecções por Roseolovirus/diagnóstico , Anticorpos Antivirais/sangue , Humanos , Imunoglobulina G/sangue , Federação Russa
19.
Klin Lab Diagn ; 64(9): 553-559, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31610108

RESUMO

The real epidemiological impact of Spotted Fever Group rickettsioses including Siberian tick-borne typhus (STT) in Russia is not sufficiently studied. One of the reasons is the actual absence of either certified domestic diagnostic kits or the evidence for using foreign test kits for laboratory verification of this group of tick-borne infections in medical practice. Objective of our study was to study the diagnostic accuracy of the ELISA test system based on Rickettsia conorii antigens for serological verification of STT. The ROC analysis was performed and operational characteristics (sensitivity, specificity, accuracy, likelihood ratio of positive and negative results) of the STT serological verification test to identify IgM to rickettsia at different times from the onset of the disease using a test system to detect antibodies to Rickettsia conorii were calculated based on the results of a survey of two groups of patients comparable by gender and age (34 patients with pathognomonic signs of STT and 76 clinically healthy people). It was found that the detection of IgM antibodies to rickettsia using the Rickettsia conorii IgM/IgG ELISA test system (Vircell) allows the disease to be verified 10-14 days after the onset of clinical symptoms in 72% (56-88%) of STT patients. We recommend the interpretation of results of the test system "Rickettsia conorii ELISA IgM/IgG" for serological verification of STT which differ from the manufacturer's recommendations regarding verification of Mediterranean fever caused by R. conorii in the following way: the diagnosis of STT should be considered laboratory confirmed when the index of IgM antibodies (IAT) exceeds 8.0; if the IAT is less than 5.0 then a repeated examination of the patient after 10-14 days will be necessary; if the IAT is in the range of 5.0-8.0 then the sample should be re-examined and / or the patient should be examined after 10-14 days. The use of the test system "Rickettsia conorii ELISA IgM / IgG" is promising for laboratory diagnosis and seroepidemiological studies of Spotted Fever Group rickettsioses in Russia.


Assuntos
Anticorpos Antibacterianos/sangue , Ensaio de Imunoadsorção Enzimática/normas , Infecções por Rickettsia/diagnóstico , Rickettsia conorii , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Federação Russa
20.
Medicine (Baltimore) ; 98(39): e17231, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31574833

RESUMO

Plasma levels of soluble PD-L1 (sPD-L1) have been reported to be an independent prognostic factor in many malignant tumors. The expression of sPD-L1 in nasopharyngeal carcinoma (NPC) has not been reported. The purpose of this study was to evaluate the expression of sPD-L1 and analyze its correlation with clinical characteristics in patients with NPC.Thirty-five patients with stage I-IVa NPC were included. Plasma samples were obtained pretreatment. The sPD-L1 concentrations were measured by enzyme-linked immunosorbent assay (ELISA). The correlations of sPD-L1 expression with clinical parameters and laboratory data were analyzed.sPD-L1 was detected in 35 plasma samples, the mean sPD-L1 concentration was 45.47 pg/ml. sPD-L1 was significantly higher in stage III-IVa (50.76 ±â€Š28.15 pg/ml) compared to stage I-II (19.87 ±â€Š11.38 pg/ml) (t = 2.618, P = .013). sPD-L1 was also higher in stage N2-3 (52.03 ±â€Š28.98 pg/ml) than that in N0-1 (32.88 ±â€Š23.75 pg/ml) (t = 2.096, P = .046). Univariate analysis identified that sPD-L1 level positively correlated with clinical stage (r = 0.495, P = .002) and N stage (r = 0.34, P = .046). Multivariate analysis showed the clinical stage was an independent factor affecting sPD-L1 expression.This is the first report to detect sPD-L1 in NPC. The study indicated sPD-L1 is quantifiable, convenient and easy to obtain. sPD-L1 may serve as a useful biomarker for evaluating tumor progression and therapeutic efficacy of NPC.


Assuntos
Antígeno B7-H1/sangue , Carcinoma Nasofaríngeo/sangue , Neoplasias Nasofaríngeas/sangue , Adolescente , Adulto , Idoso , Biomarcadores Tumorais/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Carcinoma Nasofaríngeo/patologia , Neoplasias Nasofaríngeas/patologia , Estadiamento de Neoplasias , Prognóstico , Adulto Jovem
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA