11 resultados para Biochemical and molecularcharacterization

em Universidade Federal do Rio Grande do Norte(UFRN)


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American visceral leishmaniasis is a zoonosis caused by Leishmania infantum and transmitted by the bite of the sand flies Lutzomia longipalpis.The main domestic reservoir is the dog, while foxes and opposums are the known wild reservoirs. However, identification of natural infections with L. infantum in rodents appears for need of investigating the participation of these rodents how source of infection of the parasite. In the present work the Leishmania infantum infection was investigated in rodents captured in Rio Grande do Norte, aiming at to offer subsidies to the understanding of the epidemic chains of LVA in the State. Thirteen Galea spixii were distributed in four groups, being G1 the group control with four animals and the others, G2, G3 and G4, with three animals each. Those animals were intraperitoneally inoculated with 107 promastigotas of L. infantum and accompanied for, respectively, 30, 90 and 180 days. Weekly the animals were monitored as for the corporal weight and rectal temperature. At the end of each stipulated period the animals were killed. Blood were used for determination of the parameters biochemical and haematological, PCR, ELISA, microscopic examination and cultivation in NNN medium. Liver, spleen and lymph node were used in Giemsa-stained impression and cultivation in NNN medium. Liver and spleen fragments were still used in PCR and histopathological, respectively. At the same time 79 rodents of the species Rattus rattus, Bolomys lasiurus, Oligoryzomys nigripis, Oryzomys subflavus and Trichomys apereoides were captured in the Municipal districts of Brejinho, Campo Grande, Coronel Ezequiel, Passa e Fica and Vázea for identification of natural infection with L. infantum. Evidence of infection was checked by direct examination of Giemsa-stained impression of liver, spleen and blood and culture of these tissues in NNN medium. Antibodies were researched by ELISA. They were not found differences among the weigh corporal final, rectal temperature and biochemical and haematological parameters of the Galea spixii controls and infected. The rectal temperature of the animals varied from 36OC to 40OC. For the first time values of the haematocrit (33,6% to 42,8%), hemoglobin (10,2 to 14,5g/dl), erythrocyts number (4,67x106 to 6,90x106/mm3), total leukocytes (0,9x103 to 9,2x103/mm3), platelets (49x103 to 509x103/mm3) total proteins (1,56 to 6,06 g/dl), albumin (1,34 to 3,05 g/dl) and globulins (0,20 to 3,01 g/dl) of the Galea spixii were determined. The lymphocytes were the most abundant leucocytes. Infection for L. infantum was diagnosed in two animals euthanasied 180 days after the infection. In one of the animals was also identified antibodies anti-Leishmania. The parasite was not found in none of the five other species of rodents captured. Galea spixii are resistant to the infection for L. infantum and they are not good models for the study for visceral leishmaniose, although they can act as infection sources. More studies are necessary to determine the paper of the rodents in the epidemic chain of transmission of the visceral leishmaniose in the State of Rio Grande do Norte

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Berberine is an alkaloid used as a fluorochrome in the identification of heparin and DNA. Enerback, 1974, described the technique used until today to study granules rich in heparin of vertebrate mast cells. Santos et al., 2003, studied mast cells of the mollusk Anomalocardia brasiliana using biochemical and histological analysis. This work used the fluorescent dye berberine technique to improve characterization of these cells. Mollusk organs (ctenidium and mantle) were processed with routine histological techniques. Tissue sections were treated with berberine 0,02% in redistilled water acidified to pH 4, by the addition of citric acid for 20 minutes. The visualization was made through fluorescence microscopy with ultraviolet region emission. The mast cell fluorescence had a strong yellow color, where cell nuclei appeared more greenish. This result was very similar to the ones reported before. Mast cells are location at the epithelium surface is the same in both organs, mantle and ctenidium. The fluorescence was easily observed in the granules. Therefore, this technique showed to be good and sensitive to study mast cell of invertebrates

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Compounds derived from fungi has been the subject of many studies in order to broaden the knowledge of their bioactive potential. Polysaccharides from Caripia montagnei have been described to possess anti-inflammatory and antioxidant properties. In this study, glucans extracted from Caripia montagnei mushroom were chemically characterized and their effects evaluated at different doses and intervals of treatment. It was also described their action on colonic injury in the model of colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS), and its action on cells of the human colon carcinoma (HT-29). Compounds extracted of C. montagnei contain high level of carbohydrates (96%), low content of phenolic compounds (1.5%) and low contamination with proteins (2.5%). The (FT-IR) and (NMR) analysis showed that polysaccharides from this species of mushroom are composed of α- and β-glucans. The colonic damage was evaluated by macroscopic, histological, biochemical and immunologic analyses. The results showed a reduction of colonic lesions in all groups treated with the glucans of Caripia montagnei (GCM). GCM significantly reduced the levels of IL-6 (50 and 75 mg/kg, p < 0.05), a major inflammatory cytokine. Biochemical analyses showed that such glucans acted on reducing levels of alkaline phosphatase (75 mg/kg, p < 0.01), nitric oxide (p < 0.001), and myeloperoxidase (p < 0.001). These results were confirmed microscopically by the reduction of cellular infiltration. The increase of catalase activity suggest a protective effect of GCM on colonic tissue, confirming their anti-inflammatory potential. GCM displayed cytostatic activity against HT-29 cells, causing accumulation of cells in G1 phase, blocking the cycle cell progression. Those glucans also showed ability to modulate the adhesion of HT-29 cells to Matrigel® and reduced the oxidative stress. The antiproliferative activity against HT-29 cells displayed by GCM (p <0.001) can be attributed to its cytostatic activity and induction of apoptosis by GCM

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Sugarcane (Saccharum spp.) is a plant from Poaceae family that has an impressive ability to accumulate sucrose in the stalk, making it a significant component of the economy of many countries. About 100 countries produce sugarcane in an area of 22 million hectares worldwide. For this reason, many studies have been done using sugarcane as a plant model in order to improve production. A change in gravity may be one kind of abiotic stress, since it generates rapid responses after stimulation. In this work we decided to investigate the possible morphophysiological, biochemical and molecular changes resulting from microgravity. Here, we present the contributions of an experiment where sugarcane plants were submitted to microgravity flight using a vehicle VSB-30, a sounding rocket developed by Aeronautics and Space Institute teams, in cooperation with the German Space Agency. Sugarcane plants with 10 days older were submitted to a period of six minutes of microgravity using the VSB-30 rocket. The morphophysiological analyses of roots and leaves showed that plants submitted to the flight showed changes in the conduction tissues, irregular pattern of arrangement of vascular bundles and thickening of the cell walls, among other anatomical changes that indicate that the morphology of the plants was substantially influenced by gravitational stimulation, besides the accumulation of hydrogen peroxide, an important signaling molecule in stress conditions. We carried out RNA extraction and sequencing using Illumina platform. Plants subjected to microgravity also showed changes in enzyme activity. It was observed an increased in superoxide dismutase activity in leaves and a decreased in its activity in roots as well as for ascorbate peroxidase activity. Thus, it was concluded that the changes in gravity were perceived by plants, and that microgravity environment triggered changes associated with a reactive oxygen specie signaling process. This work has helped the understanding of how the gravity affects the structural organization of the plants, by comparing the anatomy of plants subjected to microgravity and plants grown in 1g gravity

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The seeds are excellent sources of proteinase inhibitors and have been highlighted owing to various applications. Among these applications are those in effect on food intake and weight gain that stand out because of the increasing number of obese individuals. This study evaluated the effects of trypsin inhibitor present in the seed of tamarind (Tamarindus indica L.) reduction in weight gain, biochemical and morphological alterations in Wistar rats. For this, we partially purified a trypsin inhibitor tamarind seed. This inhibitor, ITT2 at a concentration of 25 mg / kg body weight, over a period of 14 days was able to reduce food intake in rats (n = 6) by approximately 47%, causing a reduction in weight gain approximately 70% when compared with the control group. With the evaluation of the in vivo digestibility was demonstrated that the animals lost weight due to satiety, presented by the reduction of food intake, since there were significant differences between true digestibility for the control group (90.7%) and the group treated with inhibitor (89.88%). Additionally, we checked the deeds of ITT2 on biochemical parameters (glucose, triglycerides, total cholesterol, high-density lipoprotein, low-density lipoprotein, glutamic-pyruvic transaminase, glutamic oxaloacetic transaminase, gamma glutamyl transferase albumin, globulin, total protein and C-reactive protein) and these, when assessed in the study groups showed no statistically significant variations. We also evaluate the histology of some organs, liver, stomach, intestine, and pancreas, and showed no changes. And to evaluate the effect of trypsin inhibitor on food intake due to the satiety is regulated by cholecystokinin (CCK) were measured plasma levels, and it was observed that the levels of CCK in animals receiving ITT2 were significantly higher ( 20 + 1.22) than in animals receiving only solution with casein (10.14 + 2.9) or water (5.92 + 1.15). Thus, the results indicate that the effect caused ITT2 satiety, reducing food intake, which in turn caused a reduction in weight gain in animals without causing morphological and biochemical changes, this effect caused by the elevation of plasma levels CCK

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The β-proteobacterium Chromobacterium violaceum is a Gram-negative, free-living, saprophytic and opportunistic pathogen that inhabits tropical and subtropical ecosystems among them, in soil and water of the Amazon. It has great biotechnological potential, and because of this potential, its genome was completely sequenced in 2003. Genome analysis showed that this bacterium has several genes with functions related to the ability to survive under different kinds of environmental stresses. In order to understand the physiological response of C. violaceum under oxidative stress, we applied the tool of shotgun proteomics. Thus, colonies of C. violaceum ATCC 12472 were grown in the presence and absence of 8 mM H2O2 for two hours, total proteins were extracted from bacteria, subjected to SDS-PAGE, stained and hydrolysed. The tryptic peptides generated were subjected to a linear-liquid chromatography (LC) followed by mass spectrometer (LTQ-XL-Orbitrap) to obtain quantitative and qualitative data. A shotgun proteomics allows to compare directly in complex samples, differential expression of proteins and found that in C. Violaceum, 131 proteins are expressed exclusively in the control condition, 177 proteins began to be expressed under oxidative stress and 1175 proteins have expression in both conditions. The results showed that, under the condition of oxidative stress, this bacterium changes its metabolism by increasing the expression of proteins capable of combating oxidative stress and decreasing the expression of proteins related processes bacterial growth and catabolism (transcription, translation, carbon metabolism and fatty acids). A tool with of proteomics as an approach of integrative biology provided an overview of the metabolic pathways involved in the response of C. violaceum to oxidative stress, as well as significantly amplified understanding physiological response to environmental stress. Biochemical and "in silico" assays with the hypothetical ORF CV_0868 found that this is part of an operon. Phylogenetic analysis of superoxide dismutase, protein belonging to the operon also showed that the gene is duplicated in genome of C. violaceum and the second copy was acquired through a horizontal transfer event. Possibly, not only the SOD gene but also all genes comprising this operon were obtained in the same manner. It was concluded that C. violaceum has complex, efficient and versatile mechanisms in oxidative stress response

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The genus Saccharum belongs to Poaceae family. Sugarcane has become important monocultures in Brazil due to their products: ethanol and sugar. The production may change between different regions from Brazil. This difference is related to soil, climatic conditions and temperature that promotes oxidative stress that may induce an early flowering. The aim of this work was to identify the effects of oxidative stress. In order to analyse this, sugarcane plants were submitted to oxidative stress using hydrogen peroxide. After this treatment, the oxidative stress were analyzed Then, the plant responses were analyzed under different approaches, using morphophysiological, biochemical and molecular tools. Thus, sugarcane plants were grown under controlled conditions and until two months they were subjected first to a hydroponics condition for 24 hours in order to acclimation. After this period, these plants were submitted to oxidative stresse using 0 mM, 10 mM, 20 mM and 30 mM hydrogen peroxide during 8 hours. The histomorphometric analysis allowed us to verify that both root and leaf tissues had a structural changes as it was observed by the increased in cell volume, lignin accumulation in cell walls. Besides, this observation suggested that there was a change in redox balance. Also, it was analyzed the activity of the SOD, CAT and APX enzymes. It was observed an increase in the SOD activity in roots and it was also observed a lipid peroxidation in leaves and roots. Then, in order to identify proteins that were differently expressed in this conditions it was used the proteomic tool either by bidimensional gel or by direct sequencing using the Q-TOF EZI. The results obtained with this approach identified more than 3.000 proteins with the score ranging from 100-5000 ions. Some of the proteins identified were: light Harvesting; oxygenevolving; Thioredoxin; Ftsh-like protein Pftf precusor; Luminal-binding protein; 2 cys peroxiredoxin e Lipoxygenase. All these proteins are involved in oxidative stress response, photsynthetic pathways, and some were classified hypothetical proteins and/or unknown (30% of total). Thus, our data allows us to propose that this treatment induced an oxidative stress and the plant in response changed its physiological process, it made changes in tissue, changed the redox response in order to survival to this new condition

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Heart transplantation (HT) represents one of the greatest advances in medicine over the last decades. It is indicated for patients with severe heart disease unresponsive to clinical treatment and conventional surgery, poor short-term prognosis and a 1- year mortality rate over 40%. HT has improved survival worldwide (80% in the first year, 70% in five years and 60% in ten years). However, the procedure has been associated with weight change and increased risk of secondary conditions such as diabetes, hypertension, dyslipidemia and obesity due to immunosuppressive therapy following transplantation. The objective of this study was to determine the impact of weight change on the metabolic stability of HT patients. The study was retrospective with data collected from the records of 82 adult patients (83% male; average age 45.06±12.04 years) submitted to HT between October 1997 and December 2005 at a transplantation service in Ceará (Brazil). The selected outcome variables (biopathological profile, weight and body mass index―BMI) were related to biochemical and metabolic change. The results were expressed in terms of frequency, measures of central tendency, Student s t test and Pearson s correlation coefficients. The analysis showed that following HT the average global BMI increased from 23.77±3.68kg/m2 to 25.48±3.92kg/m2 in the first year and to 28.38±4.97kg/m2 in the fifth. Overweight/obese patients (BMI ≥ 25 kg/m2) had higher average levels of glucose, total cholesterol, low-density lipoprotein and triglycerides than patients with eutrophy/malnutrition (BMI < 25 kg/m2). In conclusion, overweight/obese patients were likely to present higher average levels of glucose, triglycerides, total cholesterol and fractions than patients with eutrophy/malnutrition, indicating a direct and significant relation between nutritional status and weight change in the metabolic profile of HT patients

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Schistosomiasis is an ancient disease caused by helminth Schistosoma mansoni and is a public health problem in Brazil. The granulomatous lesion, typical of the disease, associates itself with increase in the oxidative damage through the generation of free radicals. The aim of this work was to evaluate the occurrence of changes in parameters oxidant / antioxidant that are part of the human defense system, and observe whether they would cause oxidative stress in subjects with schistosomiasis. Moreover, correlating with some biochemical and hematological parameters. Two groups were selected for study, consisting of individuals of both sexes, aged between 16 and 30 years. A control group, formed by individuals without schistosomiasis (n = 30) and a test group, formed by individuals with schistosomiasis (n = 30). The evaluation of lipid peroxidation in plasma was performed by determination of malondialdehyde and antioxidant defense by the quantification of reduced glutathione and catalase activity. For the parameters that assess oxidative stress, the results showed a decrease in the content of reduced glutathione and no change in the activity of catalase, with an increase in the value of malondialdehyde. Therefore, the data found suggest the occurrence of oxidative stress in subjects with schistosomiasis. Of the parameters that assess hepatic function, only levels of aspartate aminotransferase have been high, while there was a decrease of bilirubine. There was a significant change in the lipid profile (p <0.5), however with regard to the renal function of patients, there was a decrease in creatinine. The assessment hematological, made through hemogram and the quantification of hemoglobin, shows increase of eosinophils individuals in the group test, which can be related to the presence of the parasite. The amendments suggest the involvement of oxidative stress in the pathophysiology of this disease

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Introduction: The emergence of High Active Antiretroviral Therapy (HAART) increase the life expectancy of the persons living with HIV/AIDS (PLHIV), therefore the prolonged use cause metabolic implications and influences on body fat distribution and increase the cardiovascular diseases prevalence. Aims: Evaluate the effect of resistance training on heart rate variability, biochemical parameters and somatotype on PLHIV. Methods: Participated this study seven sedentary men, with age above 25 years old, living with HIV/AIDS, under HAART use. Were submitted a 16 week intervention with resistance training. Evaluated the heart rate variability, biochemical parameters and somatotype, before, after 8 weeks and 16 weeks, all in paired form. It was found the data normality by Shapiro-Wilk test and conducted the Anova one way combined with Tukey post hoc to samples in each evaluate moment, adopting significance level p<0,05. Also were calculated percentage change deltas. For somatotype was used the somatotype spatial distance (DES), obeying the significance value DES≥1. Results: Was found significance differences only in variable final heart rate delta 60s (p=0,01), however, is not showed changes on heart rate variability, biochemical parameters and somatotype components. Conclusion: 16 weeks of resistance training showed improvement on heart rate recovery after submaximal effort and, despite is not enough to produce significance differences on biochemical parameters and somatotype components, could be realize improvement on average value of fasting glucose and lipid profile, as well as reducing the endomorphic component

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Understanding the behavioral activities of freshwater shrimp in captivity is of paramount importance for the appropriate management of the species. In Brazil, the shrimp Macrobrachium rosenbergii is currently the most widely used species in the freshwater shrimp culture due to its high potential for cultivation and good market acceptance. Thus, the present study aimed to describe and characterize the behavioral activities of M. rosenbergii in monosex and in mixed (male and female) (manuscript 1, 2 and 3) populations and the growth performance of this species in restrictive feeding conditions and in different feeding management (manuscript 4 and 5, respectively) . Juvenile and adult shrimps were collected from ponds of the Aquaculture Station - Unidade Especializada em Ciências Agrárias - Universidade Federal do Rio Grande do Norte (UFRN), Macaíba/RN and then transferred to the Laboratório de Estudos do Comportamento do Camarão LECC (Laboratory for Shrimp Behavioral Studies) of the Universidade Federal do Rio Grande do Norte (UFRN). For each treatment , eight aquaria of 250 L (50 cm x 50 cm x 100 cm) were used in a closed recirculating water system with artificial lighting, constant aeration , continuous filtration through a biochemical and biological filter (canister filter), and fine sand as substrate . The water quality was monitored daily. The lab consisted of two rooms with artificial lighting system , controlled by a timer with dark / light cycle of 12:12 h . In manuscript 1, the behavioral categories of the species were presented through an ethogram, which described 31 behaviors, subdivided into general and agonistic behaviors. Manuscript 2 compared the behavioral profile of shrimps in male and in female monosex and mixed populations over 24 hours in laboratory. In three types (mixed, male monosex and female monosex) of populations during the light and dark phases of the 24 hour cycle, the shrimps showed higher occurrence of cleaning behavior. Manuscript 3 examined the influence of the color of the shelter on the frequency of its use and behavioral activities of shrimp in mixed, in male monosex and in female monosex populations over 24 hours. We observed that the shrimp M. rosenbergii burrow more frequently during the light phase in male monosex and mixed populations; they also tend to choose the black shelters. Female monosex populations tend to use red and orange shelters. In manuscript 4, we evaluated in laboratory the behavioral activities and growth performance of juvenile shrimps under food restriction. We observed that a mild food restriction may be used since there is no loss concerning the growth of the animals; feeding management on alternate days , compared to daily management can be financially productive both reducing labor costs and reducing the amount of feed used . Manuscript 5 evaluated the behavior of shrimps in monosex and in mixed populations, as well as the latency of reach the food according to feed offer (tray or food dispersal) . Our results indicate that animals adjust to both types of feed offer food dispersal as much as tray, but they spend more time to reach the feed when it is offered in trays (feeders). Comparing culture types (mixed, male monosex and female monosex), the latency to reach the food was lower for female monosex population. The data obtained in this study demonstrate the importance of identifying different pressures and environmental stimuli on the behavioral responses of this species. This knowledge would support management improvement to optimize the levels of animals‟ welfare, resulting in a better zootecnical performance