Free and nanoencapsulated vitamin D3 : effects on E-NTPDase and E-ADA activities in an animal model with induced arthritis.

UNLABELLED: The effect of vitamin D3 in oral solution (VD3 ) and vitamin D3 -loaded nanocapsules (NC-VD3 ) was analysed in animals with complete Freund's adjuvant (CFA) induced arthritis (AR). For this purpose, we evaluated scores for arthritis, thermal hyperalgesia and paw oedema, as well as histological analyses and measurements of the activity of the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) and ecto-adenosine deaminase (E-ADA) enzymes in rat lymphocytes. Haematological and biochemical parameters were also determined. The doses administered were 120 UI/day of VD3 and 15.84 UI/day of NC-VD3 . Fifteen days after the induction of AR, the groups were treated for 15 days with vitamin D3 . The results demonstrated that VD3 was able to reduce arthritis scores, thermal hyperalgesia and paw oedema in rats with CFA-induced arthritis. However, treatment with NC-VD3 did not reduce arthritis scores. The histological analyses showed that both formulations were able to reduce the inflammatory changes induced by CFA. The activity of E-NTPDase in rat lymphocytes was higher in the AR compared with the control group, while the activity of E-ADA was lower. This effect was reversed after the 15-day treatment. Data from this study indicates that both forms of vitamin D3 seem to contribute to decreasing the inflammatory process induced by CFA, possibly altering the activities of ectoenzymes. Copyright © 2016 John Wiley & Sons, Ltd. SIGNIFICANCE OF THE STUDY: The effects promoted by both formulations of vitamin D3 , either in oral solution or nanoencapsulated form, strongly suggests the softening of the inflammatory process induced by complete Freund's adjuvant (CFA), possibly altering the E-NTPDase and E-ADA activities. However, it is known that vitamin D has a beneficial effect on the modulation of the immune system components responsible for the inflammatory process. Moreover, the establishment of responses to treatment with vitamin D3 may provide an alternative for inhibiting the proinflammatory response, assisting in our understanding of the immunopathology of this disease and possibly improving the signs and symptoms that hinder the quality of life of patients with rheumatoid arthritis. HIGHLIGHTS: Evaluation of the effects on the E-NTPDase and E-ADA activities in an animal model of induced arthritis. Two formulations of vitamin D3 were used: form oral solution and nanoencapsulated. Vitamin D3 seems to contribute to the inflammatory process induced by CFA. Vitamin D3 possibly alters the E-NTPDase and E-ADA activities. Vitamin D3 may be an alternative supplementary treatment for chronic arthritis.

A prospective study on Aeromonas in outpatients with diarrhea in the central region of Rio Grande do Sul State

Aeromonas spp. were identified in five (2,7%) of 182 diarrheal stool cultures, A. caviae was predominant, resistant mainly to ampicillin and cephalotin. This is the first study showing the presence of Aeromonas spp. in diarrheal stools of outpatients in the central region of Rio Grande do Sul State, Brazil.

Influência dos anticoagulantes e da temperatura de armazenamento sobre os níveis sanguíneos de nitrito / Effects of anticoagulants and storage temperature on blood nitrite levels

A mensuração dos metabólitos do óxido nítrico pode ser útil para a melhor compreensão de diferentes processos fisiopatológicos. Este estudo avaliou a influência de diferentes anticoagulantes (ácido etilenodiaminotetracético [EDTA], citrato e heparina) e do armazenamento a -20ºC por quatro meses sobre os níveis de nitrito, sendo estes mensurados em amostras de soro e plasma pelo método de Griess. Os tipos de amostra utilizados (soro ou plasma) e de anticoagulante usado na coleta não influenciaram significativamente os níveis de nitrito, independentemente de as dosagens serem realizadas em amostras frescas ou naquelas mantidas a -20ºC por quatro meses.

The measurement of nitric oxide metabolites may be useful for better understanding of different physiopathological processes. Thus, the aim of this study was to evaluate the influence of different anticoagulants (EDTA, citrate and heparin) and four-month storage at -20ºC on nitrite levels. The serum and plasma samples were analyzed by using the Griess method. The type of sample (serum or plasma) or anticoagulant used in the collection did not influence on nitrite levels significantly, regardless the fact they were fresh or four-month samples stored at -20ºC.

Assessment of the nickel-albumin binding assay for diagnosis of acute coronary syndrome.

BACKGROUND: Myocardial ischemia may alter the metal binding capacity of circulating serum albumin. Thus, the aim of this study was to describe an automated method to measure ischemia-induced alterations in the binding capacity of serum albumin for exogenous nickel, and to evaluate the diagnostic characteristics of this assay for the assessment of acute coronary syndrome (ACS) in patients presenting to the emergency room (ER) with acute chest pain. METHODS: We assessed the concentrations of cardiac troponin I (cTnI), serum albumin, ischemia-modified albumin (IMA) measured by the cobalt-albumin binding assay (CABA), and by an automated nickel-albumin binding assay (NABA) in the following groups: ACS (n=63) and non-ischemic chest pain (NICP, n=26). Biochemical markers were determined in blood samples obtained from patients within 3 h of ER admission. RESULTS: cTnI, CABA and NABA concentrations were higher in ACS group in comparison to the NICP group. A significant correlation between NABA and CABA was observed (r=0.5387, p<0.001). Areas under the curve for CABA and NABA were 0.7289 and 0.7582, respectively. Both CABA and NABA have the ability to discriminate patients with ACS. However, NABA has a slightly higher ability to discriminate ACS compared with CABA. CONCLUSIONS: Patients with ACS have reduced nickel binding to human serum albumin, and NABA may have an important role as an early marker of myocardial ischemia, particularly in patients presenting to the ER with acute chest pain.

Enzymes that hydrolyze adenine nucleotides in platelets and polymorphisms in the alpha2 gene of integrin alpha2beta1 in patients with von Willebrand disease.

Von Willebrand disease (VWD) is one of the most common inherited bleeding diseases caused by a qualitative or quantitative deficiency of the von Willebrand factor (FvW). FvW is a multimeric glycoprotein synthesized by megakaryocytes and endothelial cells and it is present in the subendothelial matrix, blood plasma, platelets, and endothelium. This glycoprotein plays an important role in thrombus formation by initiating platelet adhesion to sites of injury as well as platelet aggregation. The aim of this study was to evaluate the activities of enzymes that hydrolyze adenine nucleotides in platelets, ristocetin-induced platelet aggregation (RIPA), and polymorphisms of the alpha2 gene of alpha2beta1 integrin from VWD patients. Platelet nucleoside triphosphate diphosphohydrolase (NTPDase), 5'-nucleotidase, and ecto-nucleotide pyrophosphatase/phosphodiesterase (E-NPP) activities were verified in 14 VWD patients. For RIPA determination, a final concentration of 1.25 mg/ml of ristocetin was used. Polymorphisms of the alpha2 gene were analyzed through PCR. Platelet NTPDase and E-NPP were decreased in VWD patients. 5'-Nucleotidase activity was not statistically significant between controls and VWD patients. RIPA was significantly reduced, with an allelic frequency of 78.57% for 807C in VWD patients. Our results indicated reduced platelet NTPDase and E-NPP activities which might be related to the low platelet adhesiveness. The prevalence of the 807C allele might account for the variability in bleeding in VWD.

Prevalência de Pseudomonas aeruginosa produtoras de β lactamases do tipo AMP-C em isolados clínicos de Santa Maria - RS / Prevalence of Pseudomonas aeruginosa AMP-C β lactamases in clinical isolates of Santa Maria - RS

Microbiologia proporcionar informações claras ao clínico, reportando corretamente os resultados, para que o tratamento seja eficaz e o mais seguro possível, evitando-se assim, a falha terapêutica e a resistência microbiana. As β-lactamases são enzimas que catalizam a hidrólise de ligações carbono-nitrogênio, separando a base do substrato inativando os antimicrobianos β-lactâmicos acarretando em falha terapêutica. Neste trabalho analisou-se 90 amostras de Pseudomonas aeruginosa provenientes de isolados clínicos de Santa Maria-RS. Empregou-se a técnica de disco difusão (Kirby-Bauer), com aproximação dos discos de Cefoxitina e Cefotaxima, onde31 cepas (34,4%) apresentaram positividade in vitro para a produção deβ-lactamases do tipo Amp-C. As cepas do grupo CESP (Citrobacter freundii, Enterobacter sp., Providencia sp., Serratia marcescens) apresentam resistência à Cefoxitina no antibiograma e não necessitariam de testes especiais para a detecção, uma vez que a enzima é produzida de forma constitutiva. Porém, alguns isolados de Pseudomonas aeruginosa, devido à produção de Amp-C em pequenas quantidades, podem apresentar sensibilidade in vitro à Cefoxitina, mas in vivo este antimicrobiano não terá ação ou sua ação será reduzida, ocasionando falha terapêutica.