Quantificação imunofenotípica de células T reguladoras em cães acometidos por linfoma multicêntrico submetidos ao protocolo quimioterápico chop associado ou não a um firocoxib

Pós-graduação em Medicina Veterinária - FCAV

Enzimas de biotransformação em tilápia (Oreochromis niloticus) exposta ao hormônio natural 17b-estradiol

Among the sexual hormones the estrogens are receiving major attention due to their biological activity. Such biological response is atributed to the best conformation recognized by their receptors, resulting in maximum responses. The estrogens are also considered responsible for most of disruptor´s effects caused by their presence in aquatic ecosystems. The 17β-estradiol hormone is produced by vertebrates during the reproductive phase of their lives and its presence has been detected in superficial waters. The objective of this study was to evaluate the cause-effect of tilapia exposition to the hormone 17β-estradiol through the analysis of biotransformation enzymes in liver and gills, complemented with the quantification of 17β-estradiol and estrone in water samples collected from fish ponds integrated to swine production. The present study was conducted under laboratory conditions, in a randomized experimental design with three levels of 17β-estradiol (E2) (0, 5, 15 µg L-1), with three replicates. After 7 days of exposure time, liver and gills were extracted to analyze three isoforms of cytochrome P450: EROD, BROD, PROD and the activity of Glutathione S-Transferase (GST). The results showed that the EROD activity (CYP1A), normally induced by the metabolism of aromatic compounds, did not present statistical differences among the treatments exposed to E2, what means that the hormone did not induce isoform 1A in fish under these particular experimental conditions. PROD activity was significantly altered in both concentrations, by means of 5 and 15 µg L-1, when compared to control. This result can indicate an important role of PROD on the metabolism of E2 present in water. Regarding to the BROD activity, it could be observed differences statistically significant between control and both groups of treatments. Two or more CYP isoforms can contribute to the metabolism of the same compound, what makes BROD a candidate as a next bioindicator of the exposure to E2 in aquatic ecosystems. Analysis of variance could confirm the effect of E2 statistically significant on the GST activity in liver tissues with >90% of significance (Prob>F = 0.0753). Furthermore, it was possible to observe that the values of GSTs activities in liver and gills in both, control and treatments, follow a tendency, that means, enzymatic activity in gills increase as the increasing of the activity in the liver tissues. In this study, the 17β–estradiol was found in measurable concentrations in three sampled points, and these values were similar to the findings of other authors at different locations in Brazil. In addition, those values are much higher than the minimum concentration that presented observable effects (10 ng L-1).

Efeitos da dose única de parecoxibe sobre a morte celular em rins de ratos submetidos à hemorragia aguda

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A protein kinase screen of Neurospora crassa mutant strains reveals that the SNF1 protein kinase promotes glycogen synthase phosphorylation

Glycogen functions as a carbohydrate reserve in a variety of organisms and its metabolism is highly regulated. The activities of glycogen synthase and glycogen phosphorylase, the rate-limiting enzymes of the synthesis and degradation processes, respectively, are regulated by allosteric modulation and reversible phosphorylation. To identify the protein kinases affecting glycogen metabolism in Neurospora crassa, we performed a screen of 84 serine/threonine kinase knockout strains. We identified multiple kinases that have already been described as controlling glycogen metabolism in different organisms, such as NcSNF1, NcPHO85, NcGSK3, NcPKA, PSK2 homologue and NcATG1. In addition, many hypothetical kinases have been implicated in the control of glycogen metabolism. Two kinases, NcIME-2 and NcNIMA, already functionally characterized but with no functions related to glycogen metabolism regulation, were also identified. Among the kinases identified, it is important to mention the role of NcSNF1. We showed in the present study that this kinase was implicated in glycogen synthase phosphorylation, as demonstrated by the higher levels of glycogen accumulated during growth, along with a higher glycogen synthase (GSN) ±glucose 6-phosphate activity ratio and a lesser set of phosphorylated GSN isoforms in strain Ncsnf1KO, when compared with the wild-type strain. The results led us to conclude that, in N. crassa, this kinase promotes phosphorylation of glycogen synthase either directly or indirectly, which is the opposite of what is described for Saccharomyces cerevisiae. The kinases also play a role in gene expression regulation, in that gdn, the gene encoding the debranching enzyme, was down-regulated by the proteins identified in the screen. Some kinases affected growth and development, suggesting a connection linking glycogen metabolism with cell growth and development.

Análise funcional da proteína 14-3-3 de Paracoccidioides brasiliensis e prospecção de alvos terapêuticos inibidores da interação de P. brasiliensis à pneumócitos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Vias de sinalização endotelial α2, AT1, e AT2 sobre a reatividade da aorta de ratos após consumo de etanol e exposição ao estresse, isoladamente e em associação

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Fracionamento de mercúrio em amostras de tecido muscular de peixes coletados na área de influência do AHE Jirau - Bacia do rio Madeira utilizando estratégias metaloproteômicas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Antidepressivo modula o comportamento e a expressão gênica das cadeias de miosina, serca2 e fosfolambam em ratos com insuficiência aórtica?

Introduction: Aortic insufficiency (AoI), a volume overload, is characterized by the diastolic reflux of blood from the regurgitating aorta to the left ventricle. This effect results from malfunctioning aortic cusps. The main cause of AoI in developing countries is rheumatic fever, including Brazil, and valvar degeneration in developed countries. There is a strong association between cardiovascular diseases and depression. Selective serotonin reuptake inhibitors (SSRI) are one of the most prescribed antidepressants in the world. Previous studies of our laboratory showed that the utilization of a SSRI, paroxetine, improved cardiac function in rats with sub-chronic AoI and reduced the daily ingestion of hypertonic sodium (NaCl 0,3M). Cardiovascular diseases can determine behavior changes like increase of anxiety, and it is yet unknown if AoI would determine anxiety or anhedonia, incapacity of obtaining pleasure through physical or sensorial experiences. A possible target for SSRI action could be a change in the expression of enzyme isoforms that collaborate in the contractile function of the heart muscle, like the heavy chains of myosine, the sarcoplasmatic reticulum Ca2+/ATPase (SERCA) and its regulator protein, phospholamban (PLB). Objectives: Evaluation of behavior parameters for anxiety and anhedonia state and genic expression of a-myosine, b-myosine, SERCA2a and PLB in the heart tissue of rats with subchronic AoI that received treatment with an SSRI (paroxetine) for 4 weeks. Methods: Surgery to induce AoI was performed on male Wistar rats, anxiety was evaluated by the elevated plus-maze (EPM) and state of anhedonia was tested by ingestion of 2% sucrose solution. After euthanasia the heart tissue was collected and total RNA was extracted to be analyzed by the RT-qPCR method. Results: Heart fractional shortening was preserved in rats with AoI that were treated compared to rats with AoI that were not treated. There was no statistically ...

Ovarian superstimulation using FSH combined with equine chorionic gonadotropin (eCG) upregulates mRNA encoding proteins involved with LH receptor intracellular signaling in granulosa cells from Nelore cows

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of exercise training on stress-induced vascular reactivity alterations: role of nitric oxide and prostanoids

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeitos preventivos do treinamento físico na expressão das proteínas Cox-2 e Vegf de ratos tratados com dexametasona

Dexamethasone is a synthetic glucocorticoid widely used to treat allergic and inflammatory processes. This drug is used in three main situations, are used to contain acute or chronic inflammatory processes, or like immunosuppressive drug's. In these cases the patient will receive high doses for a chronic period and, therefore, has a much greater chance of adverse side effects, such as hypertension, diabetes and dyslipidemia. Dexamethasone promotes deleterious effects on the arachidonic acid pathway, when administered in high doses, because it is a potent anti-inflammatory drug. We recently demonstrated that dexamethasone significantly reduces the protein expression of vascular endothelial growth factor (VEGF) in both skeletal muscle and heart, but the mechanisms involved remain unclear. Meanwhile, exercise has been shown to be effective against high blood pressure, diabetes and dyslipidemia, promoting, among other factors, the increase in VEGF and angiogenesis. One possible explanation for these effects would be the creation of new vessels mediated by inflammation, or by the stimulation of the formation of products of the metabolism of arachidonic acid (AA), such as prostaglandin E2 (PGE2) and VEGF, by increasing the stimulation of the enzymes cyclooxygenase 1 and 2 (COX-1 and COX-2). Little is known about the preventive effects of training on the action of dexamethasone in the arachidonic acid pathway. Therefore, the aim of this study was to determine whether aerobic exercise training, performed before and concomitant treatment with dexamethasone, was able to prevent the effects of the dexamethasone in the protein expression of COX-2 and VEGF. For this, we used young Wistar rats (n = 40) which were randomly divided into 4 groups: sedentary control (SC), sedentary and treated with dexamethasone (SD), trained control (TC) and trained and treated with dexamethasone (TD). These rats performed aerobic exercise training, 60% of maximum capacity, 5

Efeitos preventivos do exercício físico sobre as proteínas ciclooxigenase-2 (COX-2) e fator de crescimento de vasos derivado do endotélio (VEGF) no músculo cardíaco em ratos tratados com dexametasona

Dexamethasone (DEXA) is a synthetic glucocorticoid widely used in the handling of several drugs, for its proven benefits in fighting inflammation and allergies. Despite their benefits, their chronic use leads to several side effects that include changes in the body in the metabolism of carbohydrates, lipids and proteins. Moreover, being an anti-inflammatory, acts on the arachidonic acid pathway, reducing the expression of the enzyme cyclooxygenase (COX-2) and growth factor derived from the endothelium of blood vessels (VEGF) in various tissues. However, its effects on the myocardium are still uncertain. The physical training (PT), in turn, promotes effects contrary to those caused by chronic use of DEXA, however, little is known about the preventive effects of TF in the side effects of Dexa in the myocardium. Therefore, the aim of this study was to determine if the TF has the ability to prevent and/or mitigate the effects of Dexa in protein expression of COX-2 and VEGF in the myocardium. Forty animals were divided into 4 groups: sedentary control (SC), sedentary treated with Dexa (SD), trained control (TC) and Trained treated with Dexa (TD) and submitted to a protocol of physical training on the treadmill for 70 days (1 h/day-5 days per week, 60% of physical capacity) or kept sedentary. Over the past 10 days, rats were treated with Dexa (Decadron, 0.5 mg/kg per day, ip) or saline. During training the animals were weighed weekly and during treatment daily. At the end of treatment was made to measure fasting glucose levels of animals. The rats were killed with excess anesthesia and cardiac muscle was removed, weighed, homogenized, centrifuged and stored at -20° C for analysis of protein expression of VEGF and COX-2 by Western blotting technique. Treatment with dexamethasone caused a weight loss of 18% in sedentary animals and 13% in trained as well as elevated levels of fasting glucose in sedentary (88%). The TF was unable to mitigate the loss in...

Effects of electroacupuncture on experimental periodontitis in rats

Background: Acupuncture has shown the capability of modulating the immuno-inflammatory response of the host. This study aims to evaluate the effects of electroacupuncture (EA) on ligature-induced periodontitis in rats. Methods: Thirty-two animals were divided into four groups: 1) control; 2) experimental periodontitis (EP); 3) sham-treated (EP/EA-sham); and 4) treated with EA (EP/EA). For the EP groups, a ligature was placed around the right mandibular first molars at day 1. Sessions of EA or EA-sham were assigned every other day. For EA treatment, large intestine meridian points LI4 and LI11 and stomach meridian points ST36 and ST44 were used. EA-sham was performed in off-meridian points. Animals were euthanized at day 11. Histomorphometric and microtomographic analyses were performed. Immunolabeling patterns for the receptor activator of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG), and tartrate-resistant acid phosphatase (TRAP) were assessed. Expressions of interleukin (IL)-1 beta, matrix metalloproteinase (MMP)-8, IL-6, and cyclooxygenase (COX)-2 messenger RNAs (mRNAs) were evaluated by quantitative reverse transcription-polymerase chain reaction. Data were analyzed statistically (P < 0.05, analysis of variance). Results: Histomorphometric and microtomographic analyses demonstrated that group EP/EA presented reduced alveolar bone loss when compared to group EP (P < 0.05). Reduced RANKL immunolabeling and fewer TRAP-positive multinucleated cells were observed in the EA-treated group in relation to group EP. No differences were observed in OPG expression among groups. EA treatment decreased the genic expression of IL-1 beta and MMP-8 (P < 0.05), increased the mRNA expression of IL-6 (P < 0.05), and did not modify the genic expression of COX-2 in animals with EP (P > 0.05). Conclusion: It can be concluded that EA reduced periodontal tissue breakdown and the expression of some proinflammatory mediators and a proresorptive factor in EP in rats.

Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Insights into HLA-G genetics provided by worldwide haplotype diversity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)