510 resultados para Immunoassay


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Background: Hyperglycemia, insulin resistance and hyperleptinemia are some of the consequences of obesity. Gastric bypass for morbid obesity provides gastric restriction with decreased energy absorption. To confirm and extend previous reports in the literature, We evaluated the plasma glucose, serum insulin and leptin and insulin resistance of patients preoperatively and 1 and 3 months after Roux-en-Y gastric bypass (RYGBP).Methods: We determined body mass index (BMI), plasma glucose (glucose-oxidase method), serum leptin (immunoassay) and insulin (chemiluminescent immunometric assay), and insulin resistance index (IRI) by Homeostasis Model Assessment (HOMA) of 20 patients with morbid obesity both preoperatively and 1 and 3 months after RYGBP.Results: Patients showed a mean decrease in weight of 8 kg/month. Glycemia was above reference levels in 65% of the preoperative patients but dropped significantly 1 month postoperatively, serum insulin and leptin levels and the HOMA index also decreasing significantly in the same period. The percentage of patients with preoperative elevated serum insulin and leptin relative to reference levels decreased significantly following RYGBP. We also observed a weak but significant correlation between BMI and glucose, BMI and insulin, and leptin and insulin.Conclusions: the beneficial effects of bariatric surgery are already noticeable 1 month postoperatively, the reduction in insulin levels being more important for leptin reduction than decreased BMI. Leptin appeared to be subject to multifactorial control and showed a larger reduction than body weight.

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Many behavioral and biological variables of animals are expressed in the form of biological rhytms, down by the Circadian Timing System, that synchronize them with the environment from external stimuli such as light. One of them is the secretion profile of most circulating hormones regulated by the hypothalamuspirtuitary axis, which controls functions essential for the survival and reproduction of organisms. The sagüi, Callithrix jacchus, one of the most studied species about their endocrine physiology, is an appropriate subject for evaluating the profile of plasma prolactin and cortisol of adult males and females born in captivity throughout the year. Three male and two adult femelas were housed individually and subjected to natural environmental conditions over two years. Blood samples were used to measure the circulating levels of both hormones by methods radioimmunoassay (RIA) and immunoassay (ELISA), respectively. The analysis during the year of the plasmatic values of both hormones test was performed by ANOVA for repeated measures, the correlation of Spearman, and the test of Friedman and Student's t-test. The levels of prolactin in plasma were higher during the months in which there is a greater incidence of births of baby in the colony, possibly serving for modulating the expression of the behavior of parental care in both sexes. The plasma cortisol showed a lift in anticipation of the station with the highest birth rate and may be associated with the preparation of individual participation in caring for the baby, and also with the establishment of emotional bond between reproductive partners. Thus, this study shows that, despite the variations observed in the environment in which the animals live, plasma levels of prolactin and cortisol vary little throughout the year.

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Congenital Toxoplasmosis results in severe systemic disease. If mother is infected for the first time during gestation, she can infect the fetus causing substantial damage. However, relatively little is known about the seroprevalence and epidemiological and economic factors of Toxoplasmosis infection in pregnancy in the most state in northeastern Brazil and knowledge about this can be essential in determining effective and acceptable prevention strategies. Our aim was to determine the prevalence of Toxoplasmosis in pregnant woman consulted by reference Maternity Escola Januário Cicco in Natal, a city in Northeastern Brazil, which belongs to the public health system, correlating to the risk factors involved in the infection and to accomplish active Search in the Hospital of Pediatrics Profº Heriberto Bezerra of the damages caused by the Toxoplasmic infection in children up to 12 years of age. The study was conducted from March to December 2007 and sera obtained from 190 pregnant women were tested for IgM and IgG antibodies avidity to Toxoplasma by Microparticle enzyme immunoassay (Abbott AxSYM system - Abbott Laboratories, Chicago, IL, USA). Data were examined with univariate analysis. Chi-squared (x2) and Odds ratio was calculated (IC 95% p 0,05). Of these women, 126 (66,3%) had only IgG antibodies high-avidity against T. gondii; 01 (0,52%) had a IgM and IgG high-avidity antibodies against T. gondii and 63 (33,1%) have neither IgM nor IgG against T. gondii. Our studies shown that the direct contact with cats or dogs was highly associated with the Toxoplasma gondii infection (OR, 2.72, p<0.001, 95% CI 1.46 5.02). The years school (p<0,001), socioeconomic status and knowledge about the disease (both p value 0.05) also were associated with Toxoplasmosis. The pattern of risk factors for infection presents regional variations, however our data corroborate others studies in Brazil. In children up to 12 years, one case of Congenital Toxoplasmosis was just registered in seven years (2000 - 2006). There were several suggestive cases, with signs and characteristic symptoms, but that the infection was not confirmed due to lack in the researches through laboratorial and images exams that addressed that it zoonosis

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Background Mucocutaneous lesions in paracoccidioidomycosis are granulomatous and result from tissue responses to Paracoccidioides brasiliensis, the aetiological agent.Objectives and methods In this study we investigate the expression of tumour necrosis factor (TNF)-alpha, interleukin (IL)-10 and transforming growth factor (TGF)-beta 1 by immunohistochemistry in skin and mucosa lesions from patients with the chronic form of paracoccidioidomycosis, evaluated before and at day 20 of trimethoprim-sulfamethoxazole treatment. Cytokine production by peripheral blood monocytes was also studied by enzyme immunoassay.Results Intense immunostaining for TNF-alpha was detected in mononuclear cells that infiltrated granulomas in all skin and mucosa lesions before treatment simultaneously with low IL-10 granular deposits in these cells. At day 20 of treatment, there was reduced TNF-alpha and IL-10 deposition. Immunoreactive TGF-beta 1 was observed diffusely in the dermis and generally in the cytoplasm of macrophages and giant cells, before treatment, and as increased TGF-beta 1 deposits in the fibrosis area at day 20 of treatment. Peripheral blood monocytes from patients with paracoccidioidomycosis, evaluated before treatment, produced high endogenous levels of TNF-alpha, TGF-beta 1 and IL-10 in relation to healthy controls. Lipopolysaccharide-stimulated monocytes from patients secreted lower levels of TNF-alpha in both periods of evaluation while no impairment in capacity of IL-10 and TGF-beta production was observed.Conclusions Trimethoprim-sulfamethoxazole therapy was effective in decreasing fungal load in the lesions, allowing patient immune response to control the infection leading to the healing of the lesions.

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Monocytes and macrophages can produce a large repertoire of cytokines and participate in the pathogenesis of granulomatous diseases. We investigated the production of pro- and anti-inflammatory cytokines by monocytes from patients with active paracoccidioidomycosis. Peripheral blood monocytes from 37 patients and 29 healthy controls were cultivated with or without 10 mug/ml of lipopolysaccharide (LPS) for 18 h at 37 degreesC, and the cytokine levels were determined in the culture supernatants by enzyme immunoassay. The results showed that the endogenous levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta), IL-6, IL-8, IL-10 and transforming growth factor beta detected in the supernatant of patient monocytes cultivated without stimulus were significantly higher than those produced by healthy controls. These data demonstrated that monocytes from patients with active paracoccidioidomycosis produce high levels of cytokines with both inflammatory and anti-inflammatory activities. However, patient monocytes produced significantly lower TNF-alpha and IL-6 levels in response to LPS when compared to normal subjects, suggesting an impairment in their capacity to produce these cytokines after LPS stimulation. Concentrations of IL-1beta, IL-8 and IL-10 in cultures stimulated with LPS were higher in patients than in controls. These results suggest that an imbalance in the production of pro- and anti-inflammatory cytokines might be associated with the pathogenesis of paracoccidioidomycosis. (C) 2003 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

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Background: Biopharmaceutical drugs are mainly recombinant proteins produced by biotechnological tools. The patents of many biopharmaceuticals have expired, and biosimilars are thus currently being developed. Human granulocyte colony stimulating factor (hG-CSF) is a hematopoietic cytokine that acts on cells of the neutrophil lineage causing proliferation and differentiation of committed precursor cells and activation of mature neutrophils. Recombinant hG-CSF has been produced in genetically engineered Escherichia coli ( Filgrastim) and successfully used to treat cancer patients suffering from chemotherapy-induced neutropenia. Filgrastim is a 175 amino acid protein, containing an extra N-terminal methionine, which is needed for expression in E. coli. Here we describe a simple and low-cost process that is amenable to scaling-up for the production and purification of homogeneous and active recombinant hG-CSF expressed in E. coli cells.Results: Here we describe cloning of the human granulocyte colony-stimulating factor coding DNA sequence, protein expression in E. coli BL21(DE3) host cells in the absence of isopropyl-beta-D-thiogalactopyranoside ( IPTG) induction, efficient isolation and solubilization of inclusion bodies by a multi-step washing procedure, and a purification protocol using a single cationic exchange column. Characterization of homogeneous rhG-CSF by size exclusion and reverse phase chromatography showed similar yields to the standard. The immunoassay and N-terminal sequencing confirmed the identity of rhG-CSF. The biological activity assay, in vivo, showed an equivalent biological effect (109.4%) to the standard reference rhG-CSF. The homogeneous rhG-CSF protein yield was 3.2 mg of bioactive protein per liter of cell culture.Conclusion: The recombinant protein expression in the absence of IPTG induction is advantageous since cost is reduced, and the protein purification protocol using a single chromatographic step should reduce cost even further for large scale production. The physicochemical, immunological and biological analyses showed that this protocol can be useful to develop therapeutic bioproducts. In summary, the combination of different experimental strategies presented here allowed an efficient and cost-effective protocol for rhG-CSF production. These data may be of interest to biopharmaceutical companies interested in developing biosimilars and healthcare community.

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