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1.
Adv Exp Med Biol ; 1188: 149-163, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31820387

RESUMO

Antibodies are among the most frequently used tools in research and have had a profound impact on the discovery of diagnostic and therapeutic targets and the understanding of the molecular background of diseases. In particular in reverse phase protein arrays (RPPA), where there is no separation of the proteins according to molecular weight, it is crucial that antibodies are proven to be highly specific, selective, and reproducible. However, numerous studies have shown that many antibodies frequently do not recognize the protein that they are supposed to detect, that multiple antibodies do often function in one application but not in another, and that antibodies are not stable over time or between different batches. So far, no universally accepted guidelines or standardized methods for determining the validity of antibodies have been established. This chapter discusses the urgent need for antibody validation, current strategies that are used for (RPPA) antibody validation, as well as proposes a new strategy about how to report, score, and integrate antibody validation from multiple users.


Assuntos
Anticorpos , Análise Serial de Proteínas , Anticorpos/análise , Anticorpos/metabolismo , Proteínas/química , Proteínas/metabolismo
2.
An Bras Dermatol ; 94(6): 677-683, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31789253

RESUMO

BACKGROUND: Psoriasis is a skin-articular disease with unclear etiopathogenesis. It has been suggested that the disease is immune-mediated by T-lymphocytes, predominantly Th17 cells. Similar to psoriasis, geographic tongue is an inflammatory disease with participation of Th17 cells and direct correlation with psoriasis. OBJECTIVE: To investigate and compare the inflammatory responses and the Th17 pathway in psoriasis and geographic tongue. METHODS: This was a cross-sectional study with 46 participants that were categorized into three groups: (A) patients with psoriasis vulgaris; (B) patients with geographic tongue and psoriasis; (C) patients with geographic tongue without psoriasis. All patients underwent physical examination, and a skin and oral biopsy for histopathological examination and immunohistochemical analysis with anti-IL6, anti-IL17, and anti-IL23 antibodies. RESULTS: Histological analysis of all lesions showed mononuclear inflammatory infiltrate. However, moderate intensity was prevalent for the patients with geographic tongue and psoriasis and geographic tongue groups. Immunopositivity for the antibodies anti-IL6, anti-IL17, and anti-IL23 revealed cytoplasmic staining, mainly basal and parabasal, in both psoriasis and geographic tongue. Regarding IL-6, in patients with geographic tongue and psoriasis cases the staining was stronger than in patients with geographic tongue without psoriasis cases. IL-17 evidenced more pronounced and extensive staining when compared to the other analyzed interleukins. IL-23 presented similar immunopositivity for both geographic tongue and psoriasis, demonstrating that the neutrophils recruited into the epithelium were stained. STUDY LIMITATION: This study was limited by the number of cases. CONCLUSION: The inflammatory process and immunostaining of IL-6, IL-17, and IL-23 were similar in geographic tongue and psoriasis, suggesting the existence of a type of geographic tongue that represents an oral manifestation of psoriasis.


Assuntos
Glossite Migratória Benigna/patologia , Psoríase/patologia , Células Th17/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos/análise , Biópsia , Estudos Transversais , Feminino , Glossite Migratória Benigna/imunologia , Humanos , Imuno-Histoquímica , Interleucina-17/imunologia , Interleucina-23/imunologia , Interleucina-6/imunologia , Queratinócitos/patologia , Masculino , Pessoa de Meia-Idade , Psoríase/imunologia , Índice de Gravidade de Doença , Células Th17/imunologia , Adulto Jovem
4.
Forensic Sci Int ; 305: 110027, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31704515

RESUMO

Due the proteins from bone remains are highly resistant to pass of time and environmental conditions, they could tell us about the events that probably happened in the past. In the forensic and physical anthropology context, burnt bone remains are one of the most common pieces of recovered evidence and, generally, they are associated with funerary practices, criminal scenes or massive catastrophic events. In the present study, bone pieces of pigs were calcined at different calcination temperatures, and proteins were searched using biochemical, immunochemical and ultrastructure visualization under these experimentally conditions. For this purpose, it was successfully developed a non-demineralizing protein extraction method from burnt bone remains and the use of specific antibodies permitted the identification of different extracellular matrix and intracellular proteins. While collagen proteins type I and IV were identified and detected under middle and high calcination temperatures (300°C and 600°C); cytoskeletal proteins as actin, tubulin and, the microtubule associated protein Tau, were found under calcination process, even up high calcination temperatures. Under ultrastructural analysis, fibrous materials with a classical disposition of collagens were observed even at high calcination temperatures of the burnt bone remains. The protein identification and characterization in burnt bones as performed in present studies, is clearly demonstrating that using specific strategies for protein characterizations it is possible to found protein biomarkers in burnt bone remains and this strategy could be useful for forensic and anthropological purposes.


Assuntos
Osso e Ossos/química , Proteínas do Citoesqueleto/isolamento & purificação , Proteínas da Matriz Extracelular/isolamento & purificação , Fogo , Animais , Anticorpos/análise , Biomarcadores/química , Western Blotting , Técnica de Desmineralização Óssea , Osso e Ossos/patologia , Colágeno/ultraestrutura , Proteínas do Citoesqueleto/imunologia , Eletroforese , Proteínas da Matriz Extracelular/imunologia , Patologia Legal/métodos , Humanos , Microscopia Eletrônica de Varredura , Suínos , Temperatura Ambiente
5.
Chem Soc Rev ; 48(24): 5717-5751, 2019 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-31720618

RESUMO

The detection of clinically relevant disease-specific biomolecules, including nucleic acids, circulating tumor cells, proteins, antibodies, and extracellular vesicles, has been indispensable to understand their functions in disease diagnosis and prognosis. Therefore, a biosensor for the robust, ultrasensitive, and selective detection of these low-abundant biomolecules in body fluids (blood, urine, and saliva) is emerging in current clinical research. In recent years, nanomaterials, especially superparamagnetic nanomaterials, have played essential roles in biosensing due to their intrinsic magnetic, electrochemical, and optical properties. However, engineered multicomponent magnetic nanoparticle-based current biosensors that offer the advantages of excellent stability in a complex biomatrix; easy and alterable biorecognition of ligands, antibodies, and receptor molecules; and unified point-of-care integration have yet to be achieved. This review introduces the recent advances in superparamagnetic nanostructures for electrochemical and optical biosensing for disease-specific biomarkers. This review emphasizes the synthesis, biofunctionalization, and intrinsic properties of nanomaterials essential for robust, ultrasensitive biosensing. With a particular emphasis on nanostructure-based electrochemical and optical detection of disease-specific biomarkers such as nucleic acids (DNA and RNA), proteins, autoantibodies, and cells, this review also chronicles the needs and challenges of nanoarchitecture-based detection. These summaries provide further insights for researchers to inspire their future work on the development of nanostructures for integrating into biosensing and devices for a broad field of applications in analytical sensing and in clinic.


Assuntos
Técnicas Biossensoriais/métodos , Nanopartículas de Magnetita/química , Animais , Anticorpos/análise , Biomarcadores/análise , Técnicas Biossensoriais/instrumentação , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Desenho de Equipamento , Humanos , Nanopartículas de Magnetita/ultraestrutura , Nanotecnologia/métodos , Ácidos Nucleicos/análise , Proteínas/análise
6.
Transplant Proc ; 51(7): 2241-2244, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31400974

RESUMO

BACKGROUND: Accumulating evidence suggests that detection of human leukocyte antigen (HLA) antibodies by solid phase Luminex assays predicts renal allograft outcomes. However, several controversies exist regarding the interpretation, reproducibility, impact and financial feasibility of global utilization of this assay in pretransplant assessment. METHODS: We studied short-term patient-centered outcomes, medical standards of care, and financial plausibility of using Luminex-based screening for HLA antibodies in renal allograft recipients compared to outcomes in nontested patients. RESULTS: We included 1808 patients assessed for transplantation from 2011 to 2018. Luminex-tested patients had lower rates of rejection in the first post-transplant week (OR 0.36, P < .001) and lower odds of antibody-mediated rejection in the first 6 months (OR 0.4, P = .004). Forty-four patients with preformed, donor-specific antibodies were transplanted, and everolimus was introduced into our protocols for low-risk patients based on risk stratification by Luminex results. The number of tests needed to be performed to prevent 1 episode of antibody-mediated rejection in the first 6 months was 28 (P = .004), which was financially plausible. CONCLUSIONS: Routine pre-transplant assessment of HLA antibodies using Luminex assays may allow for better patient-centered, short-term graft outcomes and objective tailoring of immunosuppression at a financially plausible, cost-effective rate.


Assuntos
Anticorpos/análise , Rejeição de Enxerto/imunologia , Antígenos HLA/imunologia , Testes Imunológicos/métodos , Transplante de Rim/efeitos adversos , Anticorpos/imunologia , Análise Custo-Benefício , Estudos de Viabilidade , Feminino , Humanos , Testes Imunológicos/economia , Masculino , Pessoa de Meia-Idade , Período Pré-Operatório , Reprodutibilidade dos Testes , Transplante Homólogo
7.
J Sci Food Agric ; 99(15): 6903-6910, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31415094

RESUMO

BACKGROUND: Bacillus thuringiensis (Bt) synthesizes Cry1Ac protein, which is toxic to many lepidopteran pests, and the cry1ac gene has been expressed in several transgenic crop plants. The Cry1Ac protein has been isolated from Bt kurstaki HD73 and purified to homogeneity. Polyclonal antibodies were raised against purified Cry1Ac in rabbits and goat. Sandwich ELISA was developed for Cry1Ac using goat IgG as a coating antibody, and affinity-purified rabbit IgG as the primary antibody. RESULTS: The sensitivity of the assay was in the range of 0.47-1000 ng. It was subsequently employed in validating biological samples. Fifteen different cotton-seed samples were screened: 12 were found to be Bt positive and 3 Bt negative. The CS7 seeds showed the highest Bt content of 8.51 ± 0.45 µg g-1 , followed by CS8 (6.0 ± 0.02 µg g-1 ), CS15 (5.9 ± 0.03 µg g-1 ), CS9 (5.5 ± 0.05 µg g-1 ), and CS10 (4.83 ± 0.013 µg g-1 ). The CS5 seeds showed Bt content of 3.6 ± 0.21 µg g-1 . The F2 generation, CS6 (Kaveri seeds) showed lower Bt content (2.9 ± 0.06 µg g-1 ). The CL5 samples showed Cry1Ac content of 0.99 ± 0.009 µg g-1 . The amount of Cry1Ac protein in leaves, stem, and roots of germinated Bt cotton plants (CS10 and CS4) were 1.76 ± 0.15 µg g-1 , 1.9 ± 0.01 µg g-1 , 2.0 ± 0.1 µg g-1 , and 1.6 ± 0.15 µg g-1 , 1.9 ± 0.01 µg g-1 , and 2.0 ± 0.01 µg g-1 dry tissue, respectively. CONCLUSION: The method developed can be used for screening the expression levels of Cry1Ac in different transgenic Bt cultivars and also spurious Bt cotton seeds procured by farmers. © 2019 Society of Chemical Industry.


Assuntos
Bacillus/química , Endotoxinas/análise , Ensaio de Imunoadsorção Enzimática/métodos , Gossypium/química , Plantas Geneticamente Modificadas/química , Animais , Anticorpos/análise , Anticorpos/imunologia , Bacillus/metabolismo , Endotoxinas/imunologia , Endotoxinas/metabolismo , Gossypium/genética , Gossypium/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Coelhos , Sementes/química , Sementes/genética , Sementes/metabolismo
8.
Talanta ; 205: 120122, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31450437

RESUMO

The phenomenon of surface plasmon resonance (SPR) through optical sensors was developed from initial studies involving excitation of surface plasmons on metallic substrates. From the beginning, these optical systems have attracted increasing interest for application in different areas, ranging from physics, chemistry, and materials science to biology. Although numerous applications have been explored, the use of SPR in the development of biosensors is by far the most prominent. This review provides a brief account of fundamental aspects related to the recent applications of SPR as a tool for the development of new clinical diagnosis methods. The applications of SPR biosensors were illustrated through recent studies published in the field of neglected tropical diseases, with an emphasis on the contributions achieved in visceral leishmaniasis. It was possible to demonstrate the real benefits and the difficulties that the SPR biosensors have encountered in this important and complex system. Finally, future trends in the use of nanomaterials for the development of SPR-based portable devices for application to neglected tropical diseases have been demonstrated.


Assuntos
Técnicas Biossensoriais/métodos , Leishmaniose Visceral/diagnóstico , Ressonância de Plasmônio de Superfície/métodos , Anticorpos/análise , Anticorpos/imunologia , Humanos , Leishmania infantum/imunologia
9.
Transplant Proc ; 51(6): 1791-1795, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31301854

RESUMO

BACKGROUND: The 2013 Banff meeting updated the requirements for the diagnosis of acute/active antibody-mediated rejection (AAMR) in kidney allografts. There has been speculation that the changes lower the threshold for diagnosing AAMR, and may lead to possible unnecessary and expensive treatment. METHODS: We compared the 2013 Banff classification for AAMR to the previous 2007 Banff to determine if there was an increase in the number of patients receiving a diagnosis of AAMR and if the diagnosis affected allograft survival and post-biopsy 3-month and 6-month creatinine and eGFR values. RESULTS: A total of 212 renal allograft biopsies were compared to both 2007 and 2013 Banff classification requirements for AAMR. Ten patients (11 biopsies) met the 2007 criteria. An additional 15 patients (20 biopsies) met the 2013 criteria. These 2 groups showed no statistically significant demographic differences. By applying the 2013 Banff classification, we observed a 2.5-fold increase in the number of AAMR cases. One-year post-transplant allograft survival was higher in the 2013 group (.85 vs .55) and the 3-month and 6-month post-biopsy creatinine values were significantly lower for the 2013 group (1.6 ± .6 vs 3.3 ± 2.2, P value .01, and 1.7 ± .6 vs 3.4 ± 2.8, P value .03). The 3-month and 6-month eGFR values were higher in the 2013 group, although not statistically significant. CONCLUSIONS: These results suggest that use of Banff 2013 criteria in place of Banff 2007 may result in diagnosing milder and earlier cases of AAMR with the possibility of initiating earlier treatment and improving graft outcomes.


Assuntos
Anticorpos/análise , Rejeição de Enxerto/diagnóstico , Transplante de Rim/efeitos adversos , Escores de Disfunção Orgânica , Adulto , Aloenxertos/imunologia , Aloenxertos/patologia , Anticorpos/imunologia , Biópsia , Creatinina/análise , Feminino , Taxa de Filtração Glomerular , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/patologia , Sobrevivência de Enxerto/imunologia , Humanos , Rim/imunologia , Rim/patologia , Masculino , Pessoa de Meia-Idade , Transplante Homólogo , Resultado do Tratamento
10.
Nat Commun ; 10(1): 2907, 2019 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-31266958

RESUMO

Single-nucleus RNA-seq (snRNA-seq) enables the interrogation of cellular states in complex tissues that are challenging to dissociate or are frozen, and opens the way to human genetics studies, clinical trials, and precise cell atlases of large organs. However, such applications are currently limited by batch effects, processing, and costs. Here, we present an approach for multiplexing snRNA-seq, using sample-barcoded antibodies to uniquely label nuclei from distinct samples. Comparing human brain cortex samples profiled with or without hashing antibodies, we demonstrate that nucleus hashing does not significantly alter recovered profiles. We develop DemuxEM, a computational tool that detects inter-sample multiplets and assigns singlets to their sample of origin, and validate its accuracy using sex-specific gene expression, species-mixing and natural genetic variation. Our approach will facilitate tissue atlases of isogenic model organisms or from multiple biopsies or longitudinal samples of one donor, and large-scale perturbation screens.


Assuntos
Anticorpos/análise , Núcleo Celular/genética , Genômica/métodos , Análise de Célula Única/métodos , Idoso , Idoso de 80 Anos ou mais , Animais , Núcleo Celular/química , Núcleo Celular/metabolismo , DNA/genética , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/química , Neurônios/citologia , Neurônios/metabolismo , Córtex Pré-Frontal/química , Córtex Pré-Frontal/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
11.
Forensic Sci Int ; 301: 284-288, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31195249

RESUMO

Troponin I (TnI) is the inhibitory subunit of the troponin complex in the sarcomeric thin filament of striated muscle and plays a central role in the calcium regulation of contraction and relaxation. Vertebrate TnI has evolved into three isoforms encoded by three homologous genes: TNNI1 for slow skeletal muscle TnI, TNNI2 for fast skeletal muscle TnI and TNNI3 for cardiac TnI, which are expressed under muscle type-specific and developmental regulations in both the atrium and ventricle of the heart. Skeletal muscle TnI (both sTnI iso-forms) have been proposed as a sensitive and fast fiber-specific serum marker of skeletal muscle damage; fsTnI concentration in increased peripheral blood when fast twitch fibers were damaged. In our study we investigate if the 'Troponin I, fast skeletal muscle' can also be used as a reliable diagnostic tool in forensic practice, to perform differential diagnosis about vitality in suicide by hanging and simulated hanging (suspension of the victim after murder). We selected 8 women and 13 men, mean age 52.2 years, who died from suicidal hanging. The ligature material used for hanging was soft material in 11 cases and hard material in 10 cases. We chose cases as a control group of adults (n = 10; six women, four men, mean age 47.3 years) that died from opioid overdose (n = 2), car accident (n = 3) and sudden cardiac death (n = 5). Those deaths were characterized by their rapidity. To test the Anti-Troponin I fast skeletal muscle Antibody (Abcam clone-134,838), we used a case of a subject who died of myocardial infarction (timing infarct dated to 24-36 h prior to death). The reactions to Troponin I (namely the amount and extent of marker depletion) was scored for each section from 0 to -3: 0 = no loss of staining; -1 = minimal decrease in staining, compared to normally stained tissue; -2 = clear decrease in staining with some positivity (brown color) remaining; and -3 = no positive (brown) staining. The set of results obtained leads us to believe that the use of this antibody (Anti-Troponin I fast skeletal muscle antibody) is very promising to be able to make a certain differential diagnosis between antemortem and postmortem hangings. It should be emphasized that the present study seems to open new and promising horizons in the possibility to discriminate between suicidal hanging and simulated hanging (suspension of the victim after murder).


Assuntos
Asfixia/diagnóstico , Lesões do Pescoço/diagnóstico , Músculos do Pescoço/metabolismo , Suicídio , Troponina I/metabolismo , Anticorpos/análise , Biomarcadores/metabolismo , Estudos de Casos e Controles , Feminino , Patologia Legal/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Coloração e Rotulagem , Troponina I/imunologia
12.
Gastroenterology ; 157(3): 720-730.e2, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31175863

RESUMO

BACKGROUND & AIMS: Although pancreatic cystic lesions (PCLs) are frequently and incidentally detected, it is a challenge to determine their risk of malignancy. In immunohistochemical and enzyme-linked immunosorbent assay (ELISA) analyses of tissue and cyst fluid from pancreatic intraductal papillary mucinous neoplasms, the monoclonal antibody Das-1 identifies those at risk for malignancy with high levels of specificity and sensitivity. We aimed to validate the ability of Das-1 to identify high-risk PCLs in comparison to clinical guidelines and clinical features, using samples from a multicenter cohort. METHODS: We obtained cyst fluid samples of 169 PCLs (90 intraductal papillary mucinous neoplasms, 43 mucinous cystic neoplasms, and 36 non-mucinous cysts) from patients undergoing surgery at 4 tertiary referral centers (January 2010 through June 2017). Histology findings from surgical samples, analyzed independently and centrally re-reviewed in a blinded manner, were used as the reference standard. High-risk PCLs were those with invasive carcinomas, high-grade dysplasia, or intestinal-type intraductal papillary mucinous neoplasms with intermediate-grade dysplasia. An ELISA with Das-1 was performed in parallel using banked cyst fluid samples. We evaluated the biomarker's performance, generated area under the curve values, and conducted multivariate logistic regression using clinical and pathology features. RESULTS: The ELISA for Das-1 identified high-risk PCLs with 88% sensitivity, 99% specificity, and 95% accuracy, at a cutoff optical density value of 0.104. In 10-fold cross-validation analysis with 100 replications, Das-1 identified high-risk PCLs with 88% sensitivity and 98% specificity. The Sendai, Fukuoka, and American Gastroenterological Association guideline criteria identified high-risk PCLs with 46%, 52%, and 74% accuracy (P for comparison to Das-1 ELISA <.001). When we controlled for Das-1 in multivariate regression, main pancreatic duct dilation >5 mm (odds ratio, 14.98; 95% confidence interval, 2.63-108; P < .0012), main pancreatic duct dilation ≥1 cm (odds ratio, 47.9; 95% confidence interval, 6.39-490; P < .0001), and jaundice (odds ratio, 6.16; 95% confidence interval, 1.08-36.7; P = .0397) were significantly associated with high-risk PCLs. CONCLUSIONS: We validated the ability of an ELISA with the monoclonal antibody Das-1 to detect PCLs at risk for malignancy with high levels of sensitivity and specificity. This biomarker might be used in conjunction with clinical guidelines to identify patients at risk for malignancy.


Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos/análise , Biomarcadores Tumorais/análise , Ensaio de Imunoadsorção Enzimática , Neoplasias Císticas, Mucinosas e Serosas/química , Cisto Pancreático/química , Neoplasias Intraductais Pancreáticas/química , Neoplasias Pancreáticas/química , Adulto , Idoso , Anticorpos/imunologia , Especificidade de Anticorpos , Biomarcadores Tumorais/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Císticas, Mucinosas e Serosas/imunologia , Neoplasias Císticas, Mucinosas e Serosas/patologia , Neoplasias Císticas, Mucinosas e Serosas/cirurgia , Cisto Pancreático/imunologia , Cisto Pancreático/patologia , Cisto Pancreático/cirurgia , Neoplasias Intraductais Pancreáticas/imunologia , Neoplasias Intraductais Pancreáticas/patologia , Neoplasias Intraductais Pancreáticas/cirurgia , Neoplasias Pancreáticas/imunologia , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/cirurgia , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Medição de Risco , Estados Unidos
13.
Cardiology ; 142(3): 167-174, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31189164

RESUMO

PURPOSE: The aim of this study was to quantify the value of various clinical, laboratory, and instrumental signs in the diagnosis of myocarditis in comparison with morphological studies of the myocardium. METHODS: In 100 patients (65 men, 44.7 ± 12.5 years old) with "idiopathic" arrhythmias (n = 20) and dilated cardiomyopathy (DCM; n = 80), we performed the following: 71 endomyocardial biopsies (EMB), 13 intraoperative biopsies, 5 studies of explanted hearts, and 11 autopsies with virus investigation (real-time PCR) of the blood and myocardium. Antiheart antibodies (AHA) were also measured as well as cardiac CT (n = 45), MRI (n = 25), and coronary angiography (n = 47). The comparison group included 50 patients (25 men, 53.7 ± 11.7 years old) with noninflammatory heart diseases who underwent open heart surgery. RESULTS: Active/borderline myocarditis was diagnosed in 76.0% of the study group and in 21.6% of patients in the comparison group (p < 0.001). The myocardial viral genome was observed more frequently in patients in the comparison group than in the study group (65.0 and 40.2%; p < 0.01). We evaluated the diagnostic value of noninvasive markers of myocarditis. The panel of AHA had the greatest importance in the identification of myocarditis: sensitivity was 81.5%, and the positive and negative predictive values were 75.0 and 60.5%. This defined the diagnostic value of noninvasive markers of myocarditis and established a diagnostic algorithm providing an individual assessment of the likelihood of myocarditis development. CONCLUSION: AHA have the greatest significance in the diagnosis of latent myocarditis in patients with "idiopathic" arrhythmias and DCM. The use of a complex of noninvasive criteria allows the probability of myocarditis to be estimated and the indications for EMB to be determined.


Assuntos
Anticorpos/análise , Arritmias Cardíacas/diagnóstico , Cardiomiopatia Dilatada/diagnóstico , Miocardite/diagnóstico , Miocárdio/patologia , Adulto , Antiestreptolisina/sangue , Arritmias Cardíacas/sangue , Biópsia , Técnicas de Imagem Cardíaca , Cardiomiopatia Dilatada/sangue , Diagnóstico Diferencial , Feminino , Genoma Viral , Humanos , Masculino , Pessoa de Meia-Idade , Miocardite/sangue , Miocárdio/imunologia , Valor Preditivo dos Testes , Medição de Risco , Fatores de Risco , Federação Russa
14.
Anal Chim Acta ; 1067: 48-55, 2019 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-31047148

RESUMO

Immunoassay is a powerful technique to identify and quantify biological molecules, which base on the specificity and selectivity of antigen-antibody interaction. Impedance-based immunosensor has recently shown a great potential to provide rapid and label-free detections. However, the conventional impedance-based immunosensors rely on dedicated electrochemical measurement interface which involves expensive fabrication procedures such as gold deposition and photolithography. In this work, we propose an ultra-low-cost and high processing efficiency platform for impedance-based immunosensing. With effortless operations of direct-laser-writing, an impedance-based immunoassay can be fabricated within 5 min in standard laboratories. The as-fabricated devices have shown great stability and a high device-to-device uniformity. In order to further validate impedance sensing system's performance, finite element analysis and impedance equivalent model analysis were performed. The measured data was consistent with the simulation results. With the standard gold electrodes surface bio-functionalization procedures, the disposable immunoassay can detect anti-IgG down to 10 ng/ml.


Assuntos
Anticorpos/análise , Impedância Elétrica , Imunoensaio , Lasers , Impressão , Anticorpos/imunologia , Eletrodos , Ouro/química , Imunoensaio/economia , Imunoglobulina G/imunologia , Propriedades de Superfície
16.
Lab Chip ; 19(10): 1850-1859, 2019 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-31041434

RESUMO

Here we describe a combined method to monitor the secretion of molecules produced by single cells, followed by a method to isolate the individual cells that produced these molecules. The method is based on a self-sorting microwell chip that is connected to an activated membrane that collects the produced molecules. The produced molecules are printed by diffusion in small spots onto the membrane. The location of the printed spots can be correlated to the microwell number and the cell that produced these molecules. To demonstrate the method, we used the EpCAM antibody producing hybridoma cell line VU1D9 and a genetically engineered CHO cell-line producing Her2. VU1D9 cells produced 4.6 ± 5.6 pg (mean ± SD) of EpCAM antibody per 24 h and CHO cells 6.5 ± 8.2 pg per 24 h of Herceptin antibody.


Assuntos
Anticorpos/análise , Molécula de Adesão da Célula Epitelial/análise , Análise em Microsséries , Análise de Célula Única , Animais , Anticorpos/imunologia , Células CHO , Linhagem Celular Tumoral , Cricetulus , Molécula de Adesão da Célula Epitelial/biossíntese , Molécula de Adesão da Célula Epitelial/imunologia , Humanos , Impressão Tridimensional
17.
J Agric Food Chem ; 67(20): 5711-5719, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31042038

RESUMO

Although dicamba has long been one of the most widely used selective herbicides, some U.S. states have banned the sale and use of dicamba because of farmers complaints of drift and damage to nonresistant crops. To prevent illegal use of dicamba and allow monitoring of nonresistant crops, a rapid and sensitive method for detection of dicamba is critical. In this paper, three novel dicamba haptens with an aldehyde group were synthesized, conjugated to the carrier protein via a reductive-amination procedure and an indirect competitive chemiluminescent enzyme immunoassay (CLEIA) for dicamba was developed. The assay showed an IC50 of 0.874 ng/mL which was over 15 times lower than that of the conventional enzyme immunoassay. The immunoassay was used to quantify dicamba concentrations in field samples of soil and soybean obtained from fields sprayed with dicamba. The developed CLEIA showed an excellent correlation with LC-MS analysis in spike-and-recovery studies, as well as in real samples. The recovery of dicamba ranged from 86 to 108% in plant samples and from 105 to 107% in soil samples. Thus, this assay is a rapid and simple analytical tool for detecting and quantifying dicamba levels in environmental samples and potentially a great tool for on-site crop and field monitoring.


Assuntos
Anticorpos/análise , Dicamba/química , Haptenos/química , Herbicidas/química , Técnicas Imunoenzimáticas/métodos , Medições Luminescentes/métodos , Animais , Anticorpos/imunologia , Imunização , Técnicas Imunoenzimáticas/instrumentação , Medições Luminescentes/instrumentação , Espectrometria de Massas , Estrutura Molecular , Folhas de Planta/química , Coelhos , Poluentes do Solo/química , Soja/química , Espectrometria de Massas em Tandem
18.
PLoS One ; 14(4): e0214753, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30958840

RESUMO

The complement-like pathway of the African malaria mosquito Anopheles gambiae provides protection against infection by diverse pathogens. A functional requirement for a core set of proteins during infections by rodent and human malaria parasites, bacteria, and fungi suggests a similar mechanism operates against different pathogens. However, the extent to which the molecular mechanisms are conserved is unknown. In this study we probed the biochemical responses of complement-like pathway to challenge by the Gram-positive bacterium Staphyloccocus aureus. Western blot analysis of the hemolymph revealed that S. aureus challenge activates a TEP1 convertase-like activity and promotes the depletion of the protein SPCLIP1. S. aureus challenge did not lead to an apparent change in the abundance of the LRIM1/APL1C complex compared to challenge by the Gram-negative bacterium, Escherichia coli. Following up on this observation using a panel of LRIM1 and APL1C antibodies, we found that E. coli challenge, but not S. aureus, specifically activates a protease that cleaves the C-terminus of APL1C. Inhibitor studies in vivo and in vitro protease assays suggest that a serine protease is responsible for APL1C cleavage. This study reveals that despite different challenges converging on activation of a TEP1 convertase-like activity, the mosquito complement-like pathway also includes pathogen-specific reactions.


Assuntos
Anopheles/metabolismo , Proteínas de Insetos/metabolismo , Animais , Anticorpos/análise , Anticorpos/imunologia , Proteínas do Sistema Complemento/metabolismo , Dimerização , Escherichia coli/patogenicidade , Hemolinfa/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/imunologia , Inibidores de Proteases/farmacologia , Proteólise/efeitos dos fármacos , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Proteínas de Ligação a RNA/antagonistas & inibidores , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Serina Proteases/metabolismo , Staphylococcus aureus/patogenicidade , Especificidade por Substrato
19.
Anal Sci ; 35(8): 875-882, 2019 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-30982800

RESUMO

An antibody-based immunotherapy for methamphetamine (MA) addictive treatment is has been drawing more and more attention in recent years. However, studies about methamphetamine antibody (anti-MA) immunodetections are rare, owing to the lack of immunogenicity of small molecule MA. This study provides a simple and effective approach to develop a convenient electrochemiluminescent (ECL) immunosensor for the testing of anti-MA. In short, the synthetic holoantigen of MA is immobilized on a homemade gold nanoparticles modified electrode as the sensing host for the specific recognition and detection of anti-MA. The research suggested, under optimal experimental conditions, the ECL intensity on resultant immunosensor has a wide-linear regression toward the anti-MA quantity within the range from 0.03 to 3.07 ng with a detection limit of 2.32 pg. It responded to the dosage of anti-MA in spiked blood samples with satisfactory recovery. According to the research, the developed sensor shows promise as a portable Anti-MA fast seized device which performs quickly and offers convenience, and will be helpful for forensic identification and clinical treatment.


Assuntos
Anticorpos/análise , Anticorpos/imunologia , Técnicas Biossensoriais , Técnicas Eletroquímicas , Medições Luminescentes , Metanfetamina/imunologia , Eletrodos , Substâncias Luminescentes/química , Luminol/química , Metanfetamina/análise , Tamanho da Partícula , Propriedades de Superfície
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