Genotoxic assessment of Rubus imperialis (Rosaceae) extract in vivo and its potential chemoprevention against cyclophosphamide-induced DNA damage

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

Avaliação de sistemas lipidicos nanoestruturados utilizados como estratégia para melhorar as propriedades biofarmaceuticas do praziquantel no tratamento da esquistossomose

Pós-graduação em Nanotecnologia Farmacêutica - FCFAR

Avaliação de biomarcadores moleculares em mulheres com câncer de ovário

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Clinical, biochemical and haemathological effects in Rhamdia quelen exposed to cypermethrin

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

Reproductive toxicology and clastogenic evaluation in mice of a phytotherapeutic formulation obtained from Cinchona calisaya Weddel (Rubiaceae) used in Brazilian folk medicine as female fertility stimulant

Ethnopharmacological relevance: In Brazil, a phytotherapeutic preparation produced from a standardized tincture of Cinchona calisaya Weddel such that each mL of product contains 400 mu g of quinine, known in Portuguese as Agua Inglesa (R) (English water), is indicated by the manufacturer as a tonic, appetite stimulant, and digestive. However, this preparation has long been used in folk medicine as a female fertility stimulant. Despite its widespread use in folk medicine to stimulate female fertility, no study has been undertaken to assess the potential teratogenic and genotoxic effects of this phytotherapeutic preparation. The aim of the present study was to investigate possible toxic reproductive effects in mice caused by exposure to Agua Inglesa (R), either before mating or during the pre- and post-embryo implantation periods. The genotoxic potential was evaluated using the micronucleus assay.Material, Methods, and Results: Virgin female mice, with at least one estrous cycle evidenced by vaginal cytology, were divided into five groups of 15 individuals each (Group I - control, Group II - treated with ethanol solution at 16%, Groups III, IV and V treated with phytotherapeutic preparation at 1.5 mL/kg/day, 3.0 mL/kg/day and 4.5 mL/kg/day, respectively). After the first 28 days of treatment, females were caged individually with adult fertile males. Pregnant females continued to receive treatment for seven days (preimplantation period). Body weight was recorded weekly during treatment. Signs of toxicity (weight loss, food intake, piloerection, apathy, prostration, diarrhea, seizures, behavioral changes, and locomotion) were also observed. The females were sacrificed on the 15th day of pregnancy, uterine horns were evaluated for implantation, and the placental index was recorded. In the micronucleus test, 2000 polychromatic erythrocytes (PCE) per animal, obtained from bone marrow, were scored. Results The results showed that exposure of the females during the pre- and post-implantation periods did not significantly alter the reproductive capacity (p < 0.05); however, in higher dose (three times human dose)reduction of fetal weight was observed. There was no difference between the control and phytotherapeutic preparation (p > 0.05) in terms of the average number of micronucleated polychromatic erythrocytes.Conclusions: Although folk medicine suggests that the Agua Inglesa (R) preparation is useful as a female fertility stimulant, no such effect was confirmed in mice. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

Blood Profile and Histology in Oral Infections Associated with Diabetes

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

Physiological Response to Dolops carvalhoi (Crustacea: Branchiura) Infection by Pacu, Piaractus mesopotamicus, Subjected to Short Cycles of Food Restriction and Refeeding

Feeding strategies that reduce feed and promote compensatory growth could be an interesting tool to reduce costs in the fish production. However, fish health must be monitored to evaluate if their physiological response to adverse conditions, such as parasite infection, does not become compromised. A 12-wk growth trial was conducted to determine the physiological responses of pacu, Piaractus mesopotamicus, that were subjected to different fasting/refeeding cycles and infected with the Dolops carvalhoi. The schemes were: (i) control group fish (FD), (ii) food-restricted and controlled refeeding group (FR/Rc), and (iii) food-restricted and refeeding to satiation group (FR/Rs). After 84 d, the fish were exposed to D. carvalhoi for 30 h. The fish subjected to food restriction did not exhibit compensatory growth. Cortisol levels decreased in all groups within 30 h after infection. Glucose levels increased 6 h after the D. carvalhoi in the FR/Rs and 30 h after infection in the FD. In all of the fish groups, the hematocrit values were reduced after infection, and it was associated with a reduction in the mean corpuscular volume and erythrocytes. At 30 h after infection, the number of erythroblasts increased. The use of the feeding schemes does not indicate a failure of the pacu physiological responses.

Avaliação do potencial genotóxico e mutagênico do efluente de uma refinaria de petróleo e dos rios sob sua influência, por meio de bioensaios com Oreochromis niloticus (Perciformes, Cichlidae)

The petrochemical industry represents a sector of intense activity, in constant expansion and great economic importance for Brazil. In order to be used in various human activities petroleum needs to suffer a refinement process that, besides requiring large amounts of water, produces large amounts of wastes, which are discharged in hydric resources. Petroleum is a complex mixture mainly comprised by hydrocarbons, many of them are recognized as toxic chemicals, which are able to induce a considerable environmental pollution. Since water is an important resource for the maintenance of ecosystems and is also the final receiver of effluent discharges of the petroleum industry, there is an imminent need to monitor, constantly, the hydric resources that are influenced by this industrial activity, so that it can be ensured the environmental health. This study aimed, mainly, to analyze the quality of water samples derived from refinement process performed of the largest petroleum refinery in Brazil (REPLAN), in distinct steps of the industry treatment, and waters of rivers associated to its activity. Micronuclei and Nuclear Abnormalities tests were performed in erythrocytes of the fish species Oreochromis niloticus exposed to water samples derived from refinery use and to rivers under the influence of their effluents, in order to evaluate its possible toxic, genotoxic and mutagenic effects. The results obtained show that the treatment carried out by the refinery, during the evaluated periods, was effective, since the substances present in the water samples did not induce significant genotoxic and/or mutagenic alterations in the genetic material of the test organisms. On the other hand, when the substances present in the refinery effluent mix with the ones derived from other industrial activities developed upstream of this industry, the resultant compound, in some periods, presented genotoxic potencial, characterized by a rise of erythrocytic...

Avaliação da toxicidade da vinhaça utilizando Oreochromis niloticus (Perciformes: Cichlidae) como organismo teste

Contamination of rivers and lakes, waste disposed by infiltration into the soil as fertilizer, for example, has been highlighted around the world, since such compounds can contain toxic elements, causing changes in aquatic and/or terrestrial ecosystem. Due to the possible presence of these elements in these wastes, it is possible assess damage to organisms exposed through breaks in genetic material and mutations. In this sense, this work aimed at exploring the cytotoxic, mutagenic and genotoxic vinasse pontential, residue used as fertilizer, from the processing of sugar cane into alcohol, using fish (Oreochromis niloticus - Cichlidae) as a test organism. The individuals were exposed to different dilutions of vinasse in the water and later carried out analysis of their erythrocytes using the micronucleus test and nuclear abnormalities associated and the comet assay. In the first bioassay, tilapia exhibited 5% and 10% vinasse concentrations died after 48h. The number of micronucleated erythrocytes was statistically significant for the 1% vinasse concentration and also for nuclear abnormalities such as broken-egg and nuclear bud, when compared with negative and positive controls; were also significant lobed type erythrocytes, when compared to the negative control. The test results of the comet assay were not significant for 1% vinasse when compared to the negative control, due to the high rate of comets with damage control, which is unusual. The same was observed for changes as notched and blebbed type. In the second chemical analyzes conducted, the detection of heavy metals in the vinasse sample, charged only the presence of copper and chromium, the latter with a concentration 89 times higher than in the first test. Differentially the first bioassay, fish exposed to dilution of 5% did not die... (Complete abstract click electronic access below)

Freqüência do parvovírus suíno em fetos mumificados, abortados e natimortos detectado através da reação em cadeia pela polimerase (PCR)

Porcine parvovirus (PPV) is associated with reproductive failure and it has been found worldwide, including Brazil. Many diagnostic procedures are used for its detection, for example, immunofluorescence, HA, HI and PCR and this is an important technique because it is very specific and sensitive. In this work, the presence of PPV in fetuses from swine farms with reproductive problems was detected by PCR. All of 170 samples from aborted fetuses, mummies or stillborns were sampled by PCR with primers designed to VP2 region of PPV and c-myc (endogenous control). Only 142 samples (83,53%) were positive for c-myc and among them six samples (4,22%) were positive for PPV which were tested in HA. In this test, erythrocytes suspension 1% was used and three samples (50%) agglutinated but they presented low titer (4). For this work, porcine parvovirus was detected in the samples analyzed by PCR and HA. It is important to emphasize the use of endogenous control when material with elevated degree of autolysis is examined

Terapia transfusional em pequenos animais

Blood transfusion is a very important therapeutic and profilatic procedure. However, the most part of the transfusions in animals in Brazil are done with total blood. The transfusion therapy made in an appropriate way, with hemotherapic products, can save lives and warranties a better health to patients. In all transfusion procedures, the cross match test must be done because there are lots of different blood group types, based on erythrocytes antigen. A slow infusion rate and intensive patient monitoration help to prevent transfusion reactions, mainly hemolytic reactions

Expressão do transportador de monocarboxilato MCT1 e sua proteína auxiliar CD 147 em hemácias de equinos de salto

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

Polimorfismos GSTM1, GSTT1 e GSTP1 da enzima Glutationa S-transferase como fatores moduladores do fenótipo na anemia falciforme

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Phenotypic expression of homozygous hemoglobin S in relation to β-globin haplotypes, glutathione S-transferase polymorphisms and detoxification enzymes

Sickle cell anemia (SCA) shows a pathophysiology that involves multiple changes in sickle cell erythrocytes, vaso-occlusive episodes, hemolysis, activation of inflammatory mediators, endothelial cell dysfunction, and oxidative stress. These events complicate treatment and culminate in the development of manifestations such as anemia, pain crises and multiorgan dysfunction. The aim of this study was to evaluate, in SCA patients, oxidative stress and antioxidant capacity markers, correlating them to treatment with hydroxyurea (HU), β-globin haplotypes and glutathione S-transferase polymorphisms (GSTT1, GSTM1 and GSTP1), in comparison to a control group (CG). The study groups were composed of 48 individuals without hemoglobinopathies (CG), SCA patients treated with HU [AF (+HU), N = 13] and untreated SCA patients [AF (-HU), N = 15], after informed consent. The groups were analyzed using cytological, electrophoretic, chromatographic and molecular methods and information from medical records. The GSTM1 and GSTT1 polymorphisms were determined by multiplex PCR, while the GSTP1 polymorphism by PCR-RFLP. Biochemical parameters were measured using spectrophotometric methods [TBARS, TEAC and catalase (CAT) and GST activities] and a chromatographic method [glutathione (GSH)]. The fetal Hb (Hb F) levels observed in the SCA (+HU) group (10.9%) confirmed the already well-described pharmacological effect of HU, but the SCA (-HU) group also had high Hb F levels (6.1%), which may have been influenced by genetic factors not targeted in this study. We found a higher frequency of the Bantu haplotype (48.2%), followed by the Benin (32.1%) and also Cameroon haplotypes, rare in our population, and 19.7% of atypical haplotypes. The presence of Bantu haplotype was related to higher lipid peroxidation levels in patients, but also, it conferred a differential response to HU treatment, raising Hb F levels in 52.6% (P = 0.03). The protective effect of Hb F was confirmed, because the increase in their levels resulted in a 41.3% decrease in lipid peroxidation levels (r = -0.74, P = 0.0156). The genotypic frequency of the GST polymorphisms observed was similar to that of other studies in the Brazilian population, and its association with biochemical markers revealed a significant difference only for the GSTP1 polymorphism, where patients with genotype V/V showed higher GSH and TEAC levels (P = 0.04 and P = 0.03, respectively) compared to patients with genotype I/I. The TBARS levels were about five to eight times higher in the SCA (+HU) and SCA (-HU) groups, respectively, compared to controls, and HU produced a 35.2% decrease in lipid peroxidation levels in the SCA (+HU) group (P < 0.0001). Moreover, the SCA (+HU) group showed higher TEAC levels when compared to CG (P = 0.002). We did not find any significant difference in GST activity between the groups studied (P = 0.76), but CAT activity was about 17 and 30% lower in SCA (+HU) and SCA (-HU) groups, respectively (P < 0.00001). Plasma GSH levels were ~2 times higher in SCA patients than in the control group (P = 0.0005) and showed a positive correlation with TBARS levels, confirming its antioxidant function. HU treatment contributed to higher CAT activity and TEAC levels and lower lipid peroxidation, and its pharmacological effect showed a “haplotype-dependent” response. These findings may contribute to elucidating the potential of HU in ameliorating oxidative stress in SCA subjects.

Potential utility of melatonin as an antioxidant therapy in the management of sickle cell anemia

This study aimed to assess antioxidant effects of melatonintreatment compared to N-acetylcysteine (NAC) and to their combination in asickle cell suspension. Sickle erythrocytes were suspended in phosphate-buffered saline, pH 7.4, composing external control group. They were alsosuspended and incubated at 37°C either in the absence (experimental controlgroup) or in the presence of NAC, melatonin and their combination atconcentrations of 100 pM, 100 nM and 100 lM for 1 hr (treatment groups).The melatonin influences were evaluated by spectrophotometric [hemolysisdegree, catalase (CAT), glutathione S-transferase (GST), glutathioneperoxidase (GPx), glutathione reductase (GR), glucose-6-phosphatedehydrogenase (G6PDH), and superoxide dismutase (SOD) activities] andchromatographic methods [glutathione (GSH) and malondialdehyde (MDA)levels]. Incubation period was able to cause a rise about 64% on hemolysisdegree as well as practically doubled the lipid peroxidation levels (P < 0.01).However, almost all antioxidants tested treatments neutralized this incubationeffect observed in MDA levels. Among the antioxidant biomarkers evaluated,we observed a modulating effect of combined treatment on GPx and SODactivities (P < 0.01), which showed ~25% decrease in their activities. Inaddition, we found an antioxidant dose-dependent effect for melatonin onlipid peroxidation (r = 0.29; P = 0.03) and for combined antioxidanttreatments also on MDA levels (r = 0.37; P = 0.01) and on SOD activity(r = 0.54; P < 0.01). Hence, these findings contribute with important insightthat melatonin individually or in combination with NAC may be useful forsickle cell anemia management.