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PROGRAMA DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA (PGBiotecM)

UNIVERSIDADE FEDERAL DA PARAÍBA

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2024
Descripción
  • ARIANNE RAQUEL DE MENEZES MORAIS
  • POTENTIAL OF Morus nigra L. LEAF EXTRACT AS A EXTENDER COMPONENT FOR SPERM CRYOPRESERVATION
  • Asesor : SILDIVANE VALCACIA SILVA
  • Fecha: 31-ago-2024
  • Hora: 09:00
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  • The objective of this work was to evaluate different concentrations of Morus nigra L. leaf extract, in fruiting stages, as an additive to a dilute for sperm cryopreservation. The leaves collected from mulberry trees presented on Campus I (7° 05' S 34° 50' W) of the Federal University of Paraíba. The amount of 297 g of leaves were dried in an oven at 55 0C for 48 h; then, they were macerated and immersed in one liter of 95% ethanol for 72 h, protected from light, so that there was no change in the composition of photosensitive metabolites. After this process, filtration was carried out and the filtrate was dried in a vacuum rotary evaporator, with a water bath temperature of 55 0C, to obtain the crude extract. In qualitative phytochemical analysis, the presence of steroid and flavonoid compounds is obtained, and the absence of saponin, terpenoid, tannin and glycoside compounds. No Fourier Transform Infrared Spectroscopy (FTIR) test confirmed the presence of steroids and flavonoids in the extract. In the Quantification of Reducing Sugars (DNS Method), we obtained the presence of 34% of reducing sugars in the extract. In the Determination of Antioxidant Potential by the DPPH Method, the result was IC50=0.5848 mg/mL, which is favorable for a diluter. Next, the crude extract was diluted in four possible solvents: TRIS, physiological solution (NaCl 0.09%) and distilled water. The solvent chosen was TRIS to provide the best dilution of the extract. Then, the crude extract was diluted 4 mg/mL in TRIS, forming the mother solution, to later form the experimental groups: 1) Control Group (CG), composed of the Tris Gema dilutor (3.605 g of trishydroxymethyl- aminomethane; 2,024 g of citric acid 1,488 g of fructose 100 mL of doubledistilled water; 3) Test Group 2 (GT2), Tris Yolk and 5% blackberry extract; 4) Test Group 3 (GT3), Tris Yolk and 10% blackberry extract. The extenders were subjected to the Hydrogenion Potential (pH) test with the following results: The mother solution had pH 6.0 and the others pH 7.0. Considering the above parameters, the extract has potential for sperm cryopreservation due to the presence of flavonoids, which promote the reduction of oxidative stress, a satisfactory amount of sugars that can protect and provide energy for the cell, and also has considerable value as an antioxidant. As a final step, tests were carried out on equine sperm. The spermatozoa were subjected to refrigeration and freezing/thawing in the aforementioned experimental groups and subsequent analyzes (sperm motility and vigor, integrity and functionality of plasma membranes and sperm mitochondrial activity. The experiment was carried out with ejaculate from eight individuals and the evaluations carried out in duplicate and subsequently the averages were generated. Results were presented within acceptable post-thawing standards for analyzes regarding the integrity and functionality of plasma membranes.
  • JESSICA VIEIRA DANTAS
  • PHENOTYPIC VARIATION VERIFICATION OF Pseudomonas aeruginosa ISOLATES EXPOSED TO VETERINARY ANTIMICROBIALS.
  • Fecha: 15-mar-2024
  • Hora: 15:00
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  • The One Health perspective acknowledges the interconnection between human, animal, and environmental health. Antimicrobial resistance, a global challenge, affects humans and animals with significant impacts. This study investigated the responses of six isolates of Pseudomonas aeruginosa to formulations of veterinary antimicrobials under different environmental conditions. The research focused on the formation of biofilm and motility of these isolates. P. aeruginosa is recognized for its ability to form biofilms as an adaptive strategy against various environmental challenges, including antimicrobials. The results showed that the isolates exhibited a variety of phenotypic and behavioral characteristics. Hydrophobicity ranged from 91% to 110%, and the production of pyocyanin was 3.26 µg/L for JVD 05, 2.69 µg/L for JVD 04, and 1.46 µg/L for JVD 03. JVD 05 demonstrated an 83.3% inhibition of biofilm with the combination of 0.15/0.12 mg/mL of florfenicol and sulfamethoxazole-trimethoprim. Biofilm adherence was 92% for JVD 05 and 92.5% for JVD 07 when florfenicol was tested alone (0.30 mg/mL). With a DQO of 400 mg/L, there was a significant reduction in adherence in JVD 08 (34%) and higher adherence in JVD 07 (4.5%). JVD 08 exhibited the greatest reduction (42.8%) at a DQO of 2,000 mg/L, while JVD 07 showed the highest adherence (15.8%). At a DQO of 20,000 mg/L, JVD 08 recorded the greatest reduction (45.8%), and JVD 07 had the highest cellular adherence (11.8%). Regarding antimicrobial resistance, it was observed that at higher concentrations of sulfamethoxazole-trimethoprim (0.24 mg/mL) for isolate JVD 05, there were significant levels of resistance, especially under DQO conditions of 2,000 and 20,000 mg/L. The presence of florfenicol (0.30 mg/mL) and the absence of sulfamethoxazole-trimethoprim showed resistance compared to the condition of 20,000 mg/L DQO. However, the low DQO level, specifically 400 mg/L, did not seem to contribute to resistance. Additionally, in the presence of florfenicol (0.30 mg/mL), isolate JVD 05 experienced a significant reduction in motility after 24 hours, with reductions of 100% in swimming, 82.8% in swarming, and 77% in twitching, compared to controls. Florfenicol promoted the formation of biofilms classified as strong adherence in waters with higher organic matter load. Elevated concentrations of florfenicol resulted in a significant increase in Pseudomonas aeruginosa adherence.
  • ANDRWEY AUGUSTO GALVÃO VIANA
  • EVALUATION OF SYNTHETIC PHENAZINES IN THE BIODEGRADATION OF HEAVY OIL BY Pseudomonas aeruginosa TGC04
  • Asesor : ULRICH VASCONCELOS DA ROCHA GOMES
  • Fecha: 29-feb-2024
  • Hora: 10:00
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  • Between 2019 and 2020, 40% of Brazil's coastal areas suffered the most extensive oil spill in a tropical zone in history, causing negative impacts at all levels of the food chain, as well as marine ecosystem services. Many bacteria can be used in oil removal, but Pseudomonas aeruginosa is notoriously potential because it is capable of degrading hydrocarbons (HC) shortly after exposure to oil. Previous studies have shown that pyocyanin (PYO) and other intermediate phenazines help the HC transformation process by bacteria. This study aimed to verify the effect of phenazine methosulfate (PMS) on the biodegradation of sandy soil oil by allochthonous bioaugmentation with P. aeruginosa TGC04. Microcosms were prepared containing 50 g of fine sand and 150 g of beach sand contaminated with heavy oil. The pre-inoculum was prepared in 100 g of pasteurized sand and supplemented with 0.5% (w/w) of barley malt bagasse, to which 5 mL of the bacterial suspension was added, enriched with 10 µmol/L of PMS and incubated at 29±1°C for 10 days. Afterwards, portions of the inoculum were added to the microcosms (1:10; 1:100 and 1:1000). Total petroleum hydrocarbons (TPH) were reduced by up to ≈49%, while the 16 priority polycyclic aromatic hydrocarbons (HPAs) were reduced by between ≈37 and 56%. P. aeruginosa TGC04 preferentially degraded 4-6 ring HPA (80-89%), but 2-3 ring HPA were removed by up to ≈37%. In the presence of PMS there was a significant reduction in HC, however the highest rates of daily HC removal were observed in the 1:100 condition without PMS. Zea mays seeds were planted in the soil and plant development was observed for 15 days, determining important reductions in the plant's biophysical parameters. As a contribution, this study expands the knowledge that the hydrocarbonoclastic activity of P. aeruginosa is not increased by the addition of exogenous phenazines, but favors the removal of 4-6 ring HPA and that allochthonous bioaugmentation is a strategy of interest in the early stages of an oil spill accident.
  • ALEX SOUZA RIQUE
  • POTENTIAL OF Bixa orellana EXTRACT IN CRYOPRESERVATION OF GOAT SPERM CELLS
  • Asesor : SILDIVANE VALCACIA SILVA
  • Fecha: 29-feb-2024
  • Hora: 09:00
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  • The objective of this study was to evaluate the antioxidant potential of annatto extract added to sperm extender for the cryopreservation of goat sperm. Annatto seeds were mixed in a 5% potassium hydroxide (KOH) solution in a ratio of 1:6 (w/v), heated (70 ºC) and stirred for 1 h. Next, the solution was filtered and acidified (pH 3.8), in addition to a second filtration, using a paper filter; the extract was recovered through scraping and subjected to the following analyses: solubilization of the stock solutions with 1 mg/mL (SM1) and 10 mg/mL (SM10); qualitative phytochemical characterization of the extract; amount of reducing sugars (DNS); infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR); Subsequently, seven groups were created: TG (Control group); SM1-1% (Tris-yolk + 1% SM1); SM1-5% (Tris-yolk + 5% SM1); SM1-10% (Trisyolk + 10% SM1); SM10-1% (Tris-yolk + 1% SM10); SM10-5% (Tris-yolk + 5% SM10); SM10-10% (Tris-yolk + 10% SM10) subjected to hydrogen potential (pH) tests; antimicrobian activity. Then, approved ejaculates from four goats were homogenized, subjected to the freezing protocol and evaluated for kinetics (Computer Assisted Sperm Analysis), integrity and functionality of the plasma membrane. As results, for 100 g of annatto seeds, 7.45 g of wet crude extract were obtained; for the solubilization of the extract, no difference was observed between the solvents, so distilled water was chosen for the formulation of the stock solution; the spectra obtained in FTIR and NMR corroborate those observed in the literature, observing peaks referring to bixin, norbixin and geranylgeraniol; for phytochemical tests, a positive result was observed for the presence of steroids and glycosides. The results for flavonoids were inconclusive due to interference from the color of the extract, but confirmation of glycosides can reinforce the presence of this group. The presence of these groups is important precisely because they are linked to antioxidant capacity; The DNS showed the presence of 0.259 mg/mL of sugars in the extract. The presence of sugars in the medium is important for protection against cryoinjury and as a source of cellular energy. The pH of the media were all close to neutrality (6.5-7.0). This parameter is important for cellular behavior, reducing motility changes and avoiding pre-capacitation. The microbial activity test showed a weak effect of SM10 against Pseudomona aeruginosa TGC04. The microbial growth test of the experimental media did not show a bacteriostatic effect, and this result may be linked to a dose-dependent effect. A significant drop (p≤0.05) was observed in the SM10 10% experimental group in the parameters for progressive motility, VCL, VSL, VAP, WOB. In the SM 10 1% group, a smaller reduction in VCL and VAP parameters was observed. The results obtained for the highest concentration of extract indicate a possible cytotoxic effect, reducing sperm kinetic parameters. Based on the above, it is concluded that annatto extract has promising characteristics for use as an additive in cryopreservation sperm extenders, but demonstrate a negative effect at the highest concentration tested.
  • VITÓRIA TEREZA NEGRÃO DE ALBUQUERQUE
  • AXEN CULTURE AND NUTRITIONAL EVALUATION OF Pleurotus ostreatus var. florida, PRODUCED ON LIGNOCELLULOSIC SUBSTRATES
  • Asesor : ADNA CRISTINA BARBOSA DE SOUSA
  • Fecha: 28-feb-2024
  • Hora: 14:00
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  • Pleurotus spp. Its cultivation is easy due to its lignocellulosic characteristics. This allows a high colonization capacity in several compounds abundant in the agroindustry. Based on this, the objective of the present study was to evaluate the biological and productive parameters and to characterize from a physical-chemical point of view the basidiomas of P. ostreatus var. florida cultivated in different formulations based on bean pods without the grain, malt bagasse; sawdust and mesquite leaves. Initially, a test of the influence of light and temperature on mycelial development was carried out and then a qualitative pre-selection of 14 formulations for spawn and basidioma production tests. For this, individual substrates were evaluated for pH, reducing sugars, non-reducing sugars and total sugars, ash and soluble solids. After fruiting, biological, productive and nutritional aspects of the basidiomas and carbon (C), nitrogen (N), C/N ratio of the 7 pre-selected treatments were evaluated: T2 (VF), T5 (VF+GA), T7 ( BM+VF), T10 (BM+VF+GA), T11 (BM+FA), T13 (VF+FA) and T14 (VF+FA+GA). The evaluation of the influence of light on mycelial growth showed that in the dark (24h) and photoperiod (12h light/12h dark), the mushroom showed better vegetative development. treatment T1 (3.5) achieved the greatest growth, followed by T2 (2.96 cm) and T3 (2.7 cm). The ideal temperature for vegetative growth in the three treatments was between 20°C and 25°C. Physicochemical tests of the isolated substrates indicated that malt bagasse (BM) had 4.5% total sugars and grainless bean pods (VF) 3.62%. It was not possible to determine the total sugar content in mesquite leaves (FA) with the methodology used. The BM had 3.41% ash, VF 2.24% and FA 5.55%. Humidity was 8.09%, 6.9% 22.10% for BM, VF and FA, respectively. Brix was 1° (BM and VF) and 0.1° (FA). Eight formulations [T2 (VF), T6 (FA+GA), T7 (BM+VF), T8 (BM+FA), T10 (BM+VF+GA), T11 (BM+FA+GA); T13 (VF+FA) and T14 (VF+FA+GA)] showed density, vigor and continuous growth, therefore, they have the potential to produce P.ostreatus var. florida. Seven treatments [(T2 (VF), T5 (VF+GA), T7 (BM+VF), T10 (BM+VF+GA), T11 (BM+FA+GA); T13 (VF+FA) and T14 ( VF+FA+GA)] were selected for production and formed basidiomas characteristic of the species. The colonization time was 15.14 days and a total cycle of 22.14 days in all treatments. The yield percentage was 154.76 % (VF), 87.39% (VF+GA), 71.05% (BM+VF), 84.44% (BM+VF+GA), 71.26% (BM+FA), 111.01 % (VF+FA) and 53.27% (VF+FA +GA). The biological efficiency (EB%) was 38% (VF), 27.73% (VF+GA), 37.71% ( BM+VF), 35.55% (BM+VF+GA), 27.58% (BM+FA), 29.66% (VF+FA) and 16.38% (VF+FA+GA). The basidiomas produced in the 7 treatments had a low lipid value of 2 to 3%, a high protein content of 29 to 34% per 100g of dry mass, ash content of 3 to 4%, carbohydrates of 50 to 59% and an average caloric value of 350 Kcal. The C/N ratio of the 7 formulations was 26:1 (VF), 17:1 (VF+GA and BM+VF), 13:1 (BM+VF+GA and BM+FA), 14: 1 (VF+FA) and 15:1 (VF+FA+GA). Thus, the formulations with malt pomace, grain-free bean pods and mesquite leaves provided spawn production and development of the edible mushroom (Shimeji- white) with high nutritional value.
2023
Descripción
  • ALINE DAYANA PEREIRA DA SILVA
  • x
  • Asesor : ADNA CRISTINA BARBOSA DE SOUSA
  • Fecha: 31-mar-2023
  • Hora: 09:00
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  • x
  • KAÍQUE YAGO GERVAZIO DE LIMA
  • Supplementation with Lactiplantibacillus plantarum CNPC003 reverses depressive and anxiety-like behavior in a murine model.
  • Asesor : ULRICH VASCONCELOS DA ROCHA GOMES
  • Fecha: 28-mar-2023
  • Hora: 14:00
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  • More than seven million Brazilians are affected by Major Depressive Disorder (MDD) and anxiety disorder, disabling pathologies that generate impacts in the social and financial spheres. The study of the intestinal microbiota showed an intrinsic relationship with the Central Nervous System (CNS) where changes in this system alter the microbiota and the opposite is also true, also impacting TDM. Thus, the present study aims to investigate the impact of the modulation of the population of Lactobacillus sp. of the gut microbiota in an animal model of MDD and anxiety, thus evaluating the gut-brain axis. The research project was approved by CEUA/UFPB (n. 5467061021). Female Swiss (Mus musculus) mice (2-3 months) were used. Initially, the animals were induced to develop depressive and anxious behavior through the administration of dexamethasone DEXA (20µg/kg) and ATB solution, composed of Vancomycin (0.5 g/L), Ampicillin (1 g/L), Norflaxacin (1 g/L) and Nystatin (100.00 IU) (ad libitum). The groups were divided: Salina, Plantarum, Dexa, Dexa+PL, ATB, ATB+PL. The following behavioral tests were performed: Open field test, tail suspension test and elevated plus maze test. In addition to checking the estrous cycle phase and microbiological analysis to quantify the fecal population of Lactobacillus sp. The results show that both protocols, DEXA and ATB, managed to induce depressive-like behavior, as well as anxious-like behavior, in addition to causing a significant decrease in the fecal population of Lactobacillus sp. The administration of 200 μL of probiotic solution containing Lactiplantibacillus plantarum CNPC003, at a concentration of 109, managed to reverse the depressive and anxious state, as well as the Lactobacillus sp. of the intestinal microbiota. Treatment with the probiotic solution of Lactiplantibacillus plantarum CNPC003 reversed this behavior.
2022
Descripción
  • CHRISTIANNE EMMANUELLE ANDRADE PIRES BRILHANTE
  • EXTRACTION, CHARACTERIZATION AND APPLICATION OF ESSENTIAL OIL AND GROSS EXTRACT OF PEAR ORANGE PEEL IN BOVINE EPIDIMARY SPERMATOZOA CRYOPRESERVATION
  • Fecha: 30-sep-2022
  • Hora: 08:30
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  • Brazil is the country with the largest commercial cattle herd, and with that, there is a need to increasingly improve the efficiency of the processes involved in this sector. For this, biotechniques applied to reproduction, such as cryopreservation, are used. Seeking to avoid the damage caused by this technique, seminal extenders are used. However, the standard extenders used routinely use ingredients of animal origin, such as egg yolk and skimmed milk, which are considered difficult to standardize and present a risk of contamination. have themselves then a great interest in the development of extenders of vegetable origin. Orange is a fruit rich in secondary metabolites, such as carbohydrates and phenolic compounds, these are related to its antioxidant activity. In Brazil, the pear orange (Citrus sinensis (L.) Osbeck) is the most important citrus variety and its use by the juice industry generates tons of agricultural waste from its peel. Thus, this work sought to use the pear orange peel as a raw material for the production of the crude extract and the essential oil and to test the formulation of a seminal extender due to its antioxidant and antimicrobial characteristics. The essential oil was extracted and its chemical composition and antioxidant potential characterized. From the oil obtained, different nanoemulsions were formulated and tested for their cryoprotective potential in bovine epididymal cells subjected to freezing. Then, the raw extract of orange peel was obtained, followed by its characterization regarding its phytochemical composition and antioxidant potential. After the initial evaluation, extenders were tested, with and without the presence of egg yolk, with the addition of 10 and 20% of this extract. The results showed that the essential oil did not show antioxidant potential, as well as it was not successful in protecting sperm. As for the extract, it presented antioxidant potential, however, spermatozoa evaluations indicated that there was no difference between the standard extender (tris-yolk egg) and the tris-yolk egg added to the extract. The extract, without the addition of the yolk, was not able to cryopreserve bovine epididymal sperm. Orange peel extract can be better evaluated for its cryoprotective potential.
  • RAQUEL LIMA BOLCONTE
  • FERULIC ACID AS A POTENTIAL THERAPEUTIC IN THE TREATMENT OF OBESITY: AN INTEGRATIVE REVIEW
  • Fecha: 29-sep-2022
  • Hora: 14:00
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  • Overweight and obesity are responsible by the World Health Organization as the excessive accumulation of fat that presents to health, being for millions of deaths per year, is not limited to gender or age, becoming one of the most important concerns for global public health in the 21st century. Obesity represents an important risk factor for the development of other chronic diseases such as: diabetes, metabolic syndrome, cardiovascular diseases, unhealthy liver steatosis, degenerative diseases and more recently Covid-19. Some drugs are already used in the drug treatment of obesity, but in the long term they have several side effects. Thus, it is important to search for new drugs that are efficient in the treatment of obesity. Ferulic acid has been shown to be a potential therapeutic agent, having been previously reported for having several beneficial activities to human health, including antioxidant and anti-inflammatory activities, in addition to lowering cholesterol, which makes it an interesting object of study in the treatment of obesity. Therefore, this research aims to develop an integrative review of studies with ferulic acid in obesity. Initially, a research was carried out through a search in three databases: science direct, scielo and pubmed, in the period of 2021. For the selection of articles, the following inclusion criteria were adopted: experimental research articles with full texts, in English, between the years 2015 and 2020. In addition, the articles should present rodents as an animal model, and display in their title the descriptors: “ferulic acid obesity” and “ferulic acid fat”. The search was performed using the Boolean operator “AND” to associate the descriptors in the three databases. The exclusion criteria were based on the sucupira platform to evaluate the indexed journals published by the databases; articles with Qualis between A1 to B2, in the area of biotechnology, were chosen. In addition, articles that did not present the dose of ferulic acid used in the text were excluded. Through the search in the databases, it was possible to find 2,285 articles, but after a refinement of the studies, only 11 articles were selected. The main information of the articles was extracted and included in a qualitative synthesis presenting information and the main results of the articles. It was observed that oral supplementation of ferulic acid in rats can reverse almost all deleterious changes in animals studied; through insulin sensitization, with the aid of TNFα suppression and reduction of serine phosphorylation of IRS-1, PKCε, PTP1B, improving the lipid profile, reducing cholesterol and triglyceride levels. In addition, diabetic animals treated with ferulic acid showed increased glucose and insulin. Treatment with ferulic acid inhibits cell differentiation of adipocytes of the 3T3-L1 lineage; significantly reduces body weight gain, lipid level and adipose tissue mass in mice fed a hypercaloric diet; through downregulation of PPARy2, C/EBP-α and SREBP-1c levels. In addition, ferulic acid increases glycerol release from differentiated adipocytes; by activating lipolysis. Thus, these results suggest that ferulic acid has anti-obesity activity, being considered a therapeutic potential in the treatment of obesity and its complications.
  • MONIKE DE SOUSA
  • ANTIDIABETIC POTENTIAL OF FERULIC ACID IN AN EXPERIMENTAL MODEL – AN INTEGRATIVE REVIEW
  • Fecha: 28-sep-2022
  • Hora: 09:00
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  • Diabetes mellitus is a metabolic disorder that presents a picture of hyperglycemia as a result of deficiencies in the secretion of insulin, in its own insulin or in both, where the increase in the amount of glucose in the blood face so that the cells are deprived of energy. The most common types of diabetes are type 1 diabetes mellitus and type 2 diabetes mellitus. But there are more two clinical classes for diabetes: gestational diabetes mellitus and other specific types of diabetes. This disease continues to be a global epidemic and its numbers are growing exponentially each year. Beyond hyperglycemia, diabetes causes several complications affecting almost all tissues, over time, it can lead to serious complications. Ferulic acid is a phenolic acid found in many vegetables. Well known for its antioxidant properties, this acid also has antiinflammatory and antidiabetic potential. In order to analyze its effects, the present work was carried out through a literature review on the most recent results of the use of ferulic acid in the treatment of diabetes. For this, a search and survey of publications involving diabetes and the use of ferulic acid was carried out in the SCIELO - Scientific Electronic Library on-line, PubMed, and VHL - Virtual Health Library databases. To compose the sample of selected articles, the following inclusion factors were applied: articles in Portuguese, English or Spanish; that they were in full and portrayed the use of ferulic acid as an antidiabetic potential; of an experimental model with rats. And as exclusion factors: articles that did not evaluate the hypoglycemic activity of the substance;researches published and indexed outside the period from 2016 to 2021. The sample consisted of a total of 6 articles. The studies showed possible pathways for the antidiabetic mechanism of ferulic acid, including its inhibitory activity against peroxiredoxin 1, its effect against cellular stress and how it also acted in diabetic neuropathy. Through this research we also noticed that ferulic acid is used not only for hyperglycemic control, but also to improve and prevent some complications of diabetes. This demonstrates a wide range of options for studies of this promising substance.
  • ARIANE SUSAN SANTOS FREIRES
  • PRODUCTION AND BIOCHEMICAL CHARACTERIZATION OF PROTEASES OF FUNGI ISOLATED FROM SEMI-ARID SOIL SAMPLES FROM PERNAMBUCO
  • Fecha: 27-sep-2022
  • Hora: 14:00
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  • The northeastern semi-arid region is characterized by the frequent occurrence of prolonged droughts, low annual rainfall, high evapotranspiration rate, shallow and sandy soils. These factors together result in an environment with extreme and difficult conditions for the development of life. Soils are the habitat of a diverse range of macro and microorganisms that form a network of interrelationships capable of allowing and ensuring the maintenance of life in this place. Even with abiotic and biotic factors that interfere with the development of species, fungi are one of the groups of microorganisms that are most adaptable to environments like this, since these factors end up conferring resistance to these organisms, which use strategies such as the production of secondary metabolites to to ensure its survival, an example of this is enzymes. Therefore, this study aimed to evaluate the production capacity of filamentous fungi (Aspergillus sp., Paecilomyces sp., Penicillium sp. and Cladosporium sp.), isolated from soil samples from the semi-arid region of Pernambuco and to characterize the enzymatic extract with the best relationship between enzyme index (IE) and fungus biomass. The six isolates were obtained from the microbiological collection of LAPEEMI, were submitted to macroscopic (growth in Petri dish) and microscopic (coverslip culture) analysis, and the potential for proteolytic production in milk agar solid medium. The production capacity was tested in three fermentative processes, being them solid state fermentation (FES), which served as an initial screening for the best peptidases producers, following the experiments only four isolates with better IE and tested under submerged fermentation (FSm), in medium skimmed milk fibers, with agitation of 150 rpm, 37°C and stationary FSm, in triplicate in both cultures, for 6 days, after this time the samples were centrifuged to obtain the enzymatic extract. As a result, it is observed that there is a significant difference between the FSm cultivation method with agitation and stationary, for biomass production and enzymatic production respectively for the isolates Paecilomyces sp (0.368 and 0.602g and 80.3 and 302 U/mL), Penicillium sp . (1.756 and 2.913g; 75.7 and 177 U/ml), Aspergillus sp (2.852 and 3.759; 91 and 292.7 U/ml). The latter produced more biomass and the same enzymatic level than Paecilomyces sp. which had lower biomass production in stationary fermentation, so this isolate was selected for the biochemical characterization tests of the enzymes. The enzymatic extract of Paecilomyces sp was tested for peptidases classes, optimal temperature, thermostability and pH-optimum. Inhibitors were used for four classes of proteases: serine protease, aspartate protease, metalloprotease, cysteine proteases, of which it was observed the inhibition of only 20.3% of the proteases present, being 10% serine protease, and 10.3% in aspartate protease, demonstrating that most of the enzymes present do not belong to these classes. Enzyme activity was analyzed at different temperatures (27 to 97 °C) with an increment of 5 °C at each new temperature, showing greater activity at 47 °C. As for thermostability, the proteases of the crude extract, after being incubated at temperatures (27°C; 37°C; 47°C to 97°C), also with increments of 5°C, showed greater stability in the temperature ranges of 27 °C to 47 °C. The enzymatic extract showed two peaks of proteolytic activity at different pH, one at alkaline pH (pH 8.0) and the second peak, this one higher, at acid pH (pH 6.0). In view of the results obtained, it is concluded that the fungi studied were able to produce proteases by submerged fermentation in both methodologies using skimmed milk as a substrate, with emphasis on stationary cultivation, which skimmed milk medium can be considered a good alternative medium for production of these enzymes and that the protease extract of Paecilomyces sp. exhibits desired characteristics for industrial proteases.
  • CLOVIS QUEIROZ GOUVEIA
  • Qualification of the inhibitory activity of caffeine on motility and biofilm formation in Pseudomonas aeruginosa
  • Fecha: 23-sep-2022
  • Hora: 14:30
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  • Pseudomonas aeruginosa is considered an opportunistic microbe, resistant to several drugs, and presenting some characteristics such as: Gram-negative, non-fermenting, aerobic rod, and WHO (World Health Organization) classified it as a critical pathogen listed among 12 priority bacteria. P. aeruginosa can form biofilms, i.e., agglomerates of microorganisms, found in monolayers or multilayers, enabling the formation of a polymeric extracellular matrix (MEP). Caffeine is a substance extracted from coffee and other plants and exhibits antimicrobial activity against Gram-negative and Gram-positive bacteria. The present study aimed to evaluate the effect of caffeine on some virulence factors in two specimens of P. aeruginosa, isolate TGC04 and strain ATCC 9027 (cells in planktonic form, biofilm formation on two surfaces, and on motility (swimming, swarming, and twitching). Caffeine at 1024 μL/mL moderately inhibited biofilm formation on plastic surfaces (between 46.9% and 65%), while on the vitreous surface, there was a weakly inhibition (26.8 %), with no activity to isolate TGC04. Motility assays used caffeine at 2064 μL/mL which exhibited inhibition greater than 1 cm in swarming and twitching, with no activity in swimming, compared to the control test.
  • FRANCISCO SANDRO AURELIANO
  • CD8+ T LYMPHOCYTES IN COVID-19: ASSESMENT OF BIOMARKERS IN MILD AND SEVERE CLINICAL FORMS OF DISEASE.
  • Fecha: 12-ago-2022
  • Hora: 14:00
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  • Coronavirus Disease 2019 (COVID-19) is a highly transmissible acute respiratory disease caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARSCoV-2). The HCoVs of greatest epidemiological importance are MERS-CoV causing Middle East respiratory syndrome, SARS-CoV causing severe acute respiratory syndrome, and more recently SARS-CoV-2 causing COVID-19. SARS-CoV-2 is an RNA beta coronavirus and initially emerged in China in December 2019, where the first cases were associated with the Wuhan Seafood Wholesale Market, China. Since its emergence, cases of COVID-19 have spread across several countries, thus classifying it as a pandemic. Those infected with SARS-CoV-2 may have no symptoms or have symptoms ranging from mild to lethal. The current pandemic situation declared by the World Health Organization (WHO) in March 2020, due to the spread of SARSCoV-2, has been challenging for health authorities to find both therapeutic and preventive solutions for the disease, as well as understand its effects on the human organism from an immunological point of view. Some described mainly report leukocyte alterations, with lymphopenia and a decrease in T cells, including CD8+ T cells. Changes in the subpopulation of CD8+ T lymphocytes have increased the production of expression levels of activation, proliferation, and modulation markers indicating that there may be specific CD8+ T cell responses in patients with COVID19. Thus, the present study aimed to investigate the immunomodulatory mechanisms mediated by CD8 + T cells, correlating the clinical characteristics of patients recovered from mild and severe forms of COVID-19, in order to characterize the presence of prognostic markers. To this end, peripheral blood samples were obtained from recruited volunteers and distributed into control (CTL - n = 9), mild IgG - (n = 5), mild IgG+ (n = 6) and severe (n = 7) groups. Samples of PBMCs were obtained and incubated under 4 different conditions: unstimulated (medium), stimulated with SARSCoV-2 peptides (Pool Spike CoV-2 and Pool CoV-2) and stimulated with Staphylococcal enterotoxin B (SEB). CD8+ T cells were analyzed and classified regarding the expression of markers of activation (CD38 and CD69), proliferation (Ki67), as well as the production of the cytokine INF-ɣ and cytotoxic profile through the co-expression of granzyme B and perforin, and the degranulation marker CD107a between the control and recovered groups. Our analyzes generally demonstrate that both individuals who have mild COVID and are IgG+, and individuals who have severe clinical COVID-19 had a lower frequency of CD8+ T cells. In addition, the data also showed that the IgG- and/or IgG+ light groups showed an expressive production of activation markers CD69 and CD38, IFN – γ, and more cytotoxic CD8+ T cells. We can conclude that markers of activation, degranulation, and the pro-inflammatory cytokine IFN - γ may be possible prognostic markers for COVID-19 in the mild clinical form. This approach could guide investment in research in the area of Biotechnology, with an approach in the development of possible target therapies for these markers, or even the development of faster and more effective diagnostic tests in the prediction of serious diseases.
  • NÍCOLAS ALBUQUERQUE BARRETO
  • ANALYSIS OF THE INTERACTION BETWEEN CATIONIC 2-N-ALKYLPIRIDYLPORPHYRINS WITH PROTEINS INVOLVED IN Ca2+ DYNAMICS IN CARDIOMYOCYTES
  • Fecha: 29-jun-2022
  • Hora: 13:00
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  • Cardiovascular diseases (CVDs) are the leading cause of the death across the globe with myocardial infarction being responsible for 17.9 million of death in 2017. Factors such as sedentarism, unhealthy diet, alcoholism, changes in Ca2+ homeostasis and redox homeostasis are already well described as CVDS triggers. Among molecules with potential redox modulation capability and therapeutic potential are porphyrins and metaloporphyrins. Our research group and collaborators in 2020 described MnTE-2- PyP5+ as capable of modulating Ca2+ in vivo and in vitro, but this activity was not assigned to its redox modulation capability. Thus, the main goal of this work was to analyze trough molecular docking how porphyrins interact with proteins involved in the excitation-contraction coupling. CaVAb a Calcium channel with bacterial origin and a complex of cardiac troponin C with a fragment from cardiac troponin I were used. It was observed that the two used porphyrins meso-tetrakis (N-methylpiridin-2- yl)porphyrin and meso-tetrakis(N-ethylpiridin-2-yl)porphyrin obtained good affinity values for CaVAb, both in blind docking protocol and in known active site binding. As for troponin C, it was observed a major interaction with the N domain of the protein, different from the known binding site for this structure. Therefore, it was possible to conclude that the used porphyrins, based on the values of affinity found have a high potential to modulate CaVAb and low potential do sensibilize the troponin C to Ca2+ Keywords: cardiomyocytes, porphyrins, molecular docking, calcium channel, troponin C
  • JOSE DE ANCHIETA DE OLIVEIRA FILHO
  • Virtual Thyroid Alpha Receptor Agonist Screening to Stimulate Cardiac Regeneration
  • Fecha: 17-mar-2022
  • Hora: 08:00
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  • Cardiovascular diseases are the ones which kill the most in the world, being ischemicdiseasesthe most representative, with approximately 15 million deaths worldwide in the year 2016alone. One of the main causes of these deaths is due to the fact that the cardiac tissuehasalow rate of proliferation and regeneration after injury, which follows scarringpathways,resulting in partial or total loss of its function. However, the ability to induce cell regenerationand proliferation by this organ via pharmacological treatment has already been demonstrated.In particular, thyroid hormones have a beneficial influence on the aforementionedevents,effects linked to the alpha thyroid receptor. Thus, in this work a virtual screeningwasperformed in search of agonists for the target in question, based on the ZINC15 database. Forthis, a physicochemical filter was used, eliminating unwanted compounds, obtaining2.291.411 desired compounds to be sorted. Then, a pharmacophoric filter was appliedthroughthe ROCS program based on triiodothyronine, selecting the candidates throughtheROCcurve and population dispersion graphs, obtaining 10.633 potentially active ligands. Then,with Molecular Docking, using the OEDocking FRED program, we selected 568 compoundsfor visual analysis. These were visually inspected based on inclusion criteria guidedbythechemical knowledge of the target. Thus, at the end of the screening we obtained 27potentiallyagonist molecules and 15 with less bioactive potential for the alpha thyroid receptor. Thus, 8candidates were selected to be selected using Molecular Dynamics and Binding FreeEnergytechniques, finally, 2 promising candidates were selected for thyroid receptor alphaagonistsfor in vitro validation.
  • FILIPE DE CASTRO PATRICIO
  • EXTRACTION AND CHARACTERIZATION OF XYLAN FROM CORN AND SUGAR CANE FOR THE PROSPECTION OF XYLANASES IN BACTERIAL ISOLATES
  • Fecha: 24-feb-2022
  • Hora: 14:00
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  • The reuse of agro-industrial residues has been the focus of several researches, mainly in the field of biotechnology, due to the biomass of these residues being rich in polymers that have a high added value, such as xylan. Xylan is the main hemicellulose, being the second most abundant polysaccharide, it has several applications such as in the food, pharmaceutical and biotechnology industry, and can be used in enzymatic prospecting using microorganisms. This study aimed to extract xylan from three agricultural residues, corn cob, bagasse and sugarcane straw for bioprospecting of xylanases using bacterial isolates from soil samples from Usina Japungu Agroindustrial S/A, located in the municipality of Santa Rita – Paraiba. As a result of the xylan extraction step from sugarcane residues, a yield of 4.62g for straw and 3.80g for bagasse was obtained, starting from 30g of residue. To characterize the xylans obtained, the Fourier Transform Infrared Spectroscopy (FT-IR) method was used, which showed the main bands present in the xylan. For xylanase prospection, 21 strains were selected from the microorganism bank of the Laboratory of Cellular and Molecular Biotechnology, among which, 16 strains grew and were used in the enzyme production assays, in agar/Luria Bertani medium, added 0.4% xylan of the corn cob, sugarcane bagasse and straw and commercial xylan (Beechwood), separately, which were incubated for 72 h at 37°C and stained with Congo red (0.1%) to observe the halo of degradation and determination of the enzymatic index (IE). Among the xylanase positive strains (11 in total), 5 strains were positive for the 4 types of xylans used and 6 strains were positive only for commercial xylan. For enzymatic production, strains JS19, which presented IE ≥ 2, in all tested media, and JS20, which presented enzyme production only in the medium containing commercial xylan, were selected. The strains were cultivated in nutrient broth added with 0.5% of industrial residue, corn cob, straw and sugarcane bagasse, and incubated in a shaker (150 rpm) at 37°C for 72 h to evaluate the enzymatic activity (U/mL), by the reducing sugar method (3,5-dinitrosalicylic acid - DNS). As a result of enzyme production, JS19 showed a better quantification, ranging from 0.74 to 0.92 (U/mL), compared to strain JS20, which ranged from 0.12 to 0.30 (U/mL). These preliminary data show that the reuse of agro-industrial residues, such as corn and sugarcane, are promising for obtaining xylans, as well as for enzymatic bioprospecting using microorganisms, such as bacteria isolated from the soil.
2021
Descripción
  • LOUISE HELENA GUIMARÃES DE OLIVEIRA
  • STUDY OF THE EFFECTS OF ULTRAVIOLET RADIATION ON THE Aedes aegypti MOSQUITO
  • Fecha: 20-jul-2021
  • Hora: 08:00
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  • Most Brazilian states are infested with Aedes aegypti, which allows the transmission of several diseases. The mosquito is the vector of several important arboviruses, such as dengue, urban yellow fever, zika and chikungunya, among other diseases, being the target of major concern for public health, the main way to combat these diseases is through the control of the vector mosquito. Several studies show the emergence of mosquitoes resistance. Due to the resistance already observed, there is a search for new tools to control this possible vector. However, there is also an environmental concern, as the use of insecticides can directly affect nature. Thus, it is necessary to search for forms of control that would not leave residues in nature. The UV radiation is already used for biological control of some organims, the study of the exposure of this ultraviolet radiation shows itself as an effective and clean way to fight the Aedes aegypti mosquito. The objective of this work was to study the ovicidal, larvicidal, pupicidal and adulticidal activities of ultraviolet radiation in Ae. aegypti. As a method, for the evaluation of ovicidal activity, eggs of Ae. aegypti were exposed and evaluated in the period of 25 days, as a result, a decrease in hatchability of 95.53% of Ae. aegypti was observed. In order to perform larvicidal activity, the larvae were exposed to different periods of light and their mortality was verified in 30, 45, 60 and 180 minutes, as a result, it was possible to observe a mortality of 27.77; 45.53; 76.67 and 96.67% of the larvae, respectively. For the evaluation of pupicidal activity, the pupae of Ae. aegypti were exposed and observed in a 24h period, as a result, it was possible to observe the mortality of 86.67% of the exposed pupae. To carry out the adulticidal activity, the mosquitoes of Ae. aegypti were exposed to ultraviolet light and were observed in 24 and 48 hours, the results showed the mortality of 86.67 and 93.33% of mosquitoes, respectively. All experiments were carried out in triplicate.
2020
Descripción
  • MARIA GABRIELLA SILVA SIDRÔNIO
  • Drug repositioning for the treatment of tuberculosis: evaluation of the antimycobacterial activity of antimalarial compounds
  • Fecha: 22-dic-2020
  • Hora: 14:00
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  • Mycobacterium tuberculosis is the pathogen that causes tuberculosis, a contagious infectious disease that spreads through the airways of infected individuals. Tuberculosis is a leading cause of death worldwide and one of the factors that contribute to this condition is the emergence of strains resistant to the recommended therapeutic regimen to treat this disease. In order to reduce the high incidence rates of tuberculosis, researchers are looking for new alternatives for the treatment of this disease. In this sense, some studies indicate that antimalarials have antimicrobial activity against strains of Mycobacterium tuberculosis. This study aimed to evaluate the activities of primaquine, chloroquine, mefloquine and tafenoquine, in strains of Mycobacterium tuberculosis from Mycobacterium smegmatis mc2155. Initially, antimalarials were incubated with Mycobacterium tuberculosis H37Ra and Mycobacterium smegmatis, in order to determine their minimum inhibitory concentrations. The activities of the test molecules were compared with each other and with drugs used in the treatment of tuberculosis. Additionally, combinatorial tests were carried out involving the antimalarial molecule with the greatest potential, tafenoquine, and drugs used in the treatment of tuberculosis. After determining the antimalarial molecules with the greatest potential, suspensions of Mycobacterium tuberculosis H37Ra were subjected to models of stress and numbness. Experiments were also carried out to determine a death curve for the antimalarial with the greatest potential in Mycobacterium tuberculosis H37Ra. Tests were also carried out in order to determine the minimum inhibitory concentration in resistant and virulent strains of Mycobacterium tuberculosis, as well as tests to determine the molecular target of the most potential antimalarial in these microorganisms. After carrying out these tests, it was possible to conclude that all test molecules showed antimicrobial activity against strains of Mycobacterium tuberculosis, as well as Mycobacterium smegmatis, and the drugs of mefloquine and tafenoquine were considered to have the greatest potential. Regarding combinatorial tests, it was possible to observe that mefloquine, when combined with tafenoquine, has a synergistic effect in Mycobacterium tuberculosis. After carrying out experiments on bacteria submitted to stress and dormancy models, it was possible to observe that the drugs of tafenoquine and mefloquine has antimicrobial activity in suspensions of Mycobacterium tuberculosis subjected to nitrosative stress and nutrient depletion. Additionally, it was possible to observe that tafenoquine, as well as mefloquine, have activity in resistant and virulent strains of Mycobacterium tuberculosis. The satisfactory activity of antimalarials in strains of Mycobacterium spp. suggests that these drugs may be considered promising candidates for the treatment of tuberculosis.
  • NÍVEA PAULA CORDEIRO DIAS
  • DIAGNOSTIC PROFILE EVALUATION Mycobacterium tuberculosis RESISTANCE TO RIFAMPICIN AND A MOBILE APPLICATION PROPOSAL FOR ASSISTANCE TO THE TREATMENT OF TUBERCULOSIS IN A HOSPITAL REFERENCE IN THE STATE OF PARAIBA
  • Fecha: 04-dic-2020
  • Hora: 09:00
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  • Tuberculosis is an infectious disease and a serious public health problem worldwide. According to the WHO in its latest report, about 10 million people contracted tuberculosis and 1.5 million people died from the disease. In 2018, about half a million people developed rifampin-resistant tuberculosis and 78% of them presented the multidrug-resistant form. In Brazil, about 76,000 new cases were reported and 4,500 deaths from the disease. The present study aimed to evaluate the diagnostic profile performed for the detection of tuberculosis by molecular and phenotypic means, resistance to rifampicin and from the observed data to propose innovation through the creation of a mobile application prototype to assist in the treatment of tuberculosis in patients seen at the referral hospital for infectious diseases in Paraíba. The research has an epidemiological, retrospective, observational, transversal, individualized character with an emphasis on quantitative data analysis and from these the development of a mobile application prototype, structured through the Adobe XD platform. The data were stored in a tabular structure and consist of numerical and categorical values, as well as time series. From May 2015 to May 2019, an average of 827 molecular diagnostic tests were performed annually as the main test for tuberculosis detection. The incidence of cases with results of sensitivity to rifampicin indicated in the Rapid Molecular Test (TRM) was higher than the cases that presented resistance; 95.52% cases with verified sensitivity and 4.47% resistance. The tests that showed a mutation of the rpoB gene varied and of these, only 18 continued with the Antimicrobial Sensitivity Test (TSA), of which 12 obtained a concordant phenotypic result in the TSA compared to the TRM, with confirmed rifampicin resistance and 6 patients presented discordant result; the phenotypic profile had a higher resistance index for isoniazid 24.07% and the highest sensitivity, 98.14% for ethambutol. The prototype entitled “TRATE TB” was developed based on the scenario created from data analysis, with visual access, handling and simulation of features such as: scheduling of reminders for medication intake, exam attachment, appointment scheduling, inventory registration medicines, generating progress reports and adhering to treatment, as well as sharing information with medical and family assistance. Even partially remotely, the “TRATE TB” application should play a considerable role in assisting patients undergoing treatment for tuberculosis at the Reference Hospital Clementino Fraga, in addition to the fact that easy access to resources in a single tool can intervene in the number of new cases, in the development of resistance to the main drugs, abandonment of treatment and cure of the patient.
  • GABRIELLY DINIZ DUARTE
  • Synthesis, characterization and in silico evaluation of heterocyclic chalcone derivatives as potential bronchodilators
  • Fecha: 16-sep-2020
  • Hora: 14:00
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  • Chalcones are molecules of high biological value due to it’s variety in biological activities. In this context, derivatives of synthetic chalcones are promising molecules in drug development of interest for both the academy and industry. Seeking to contribute to the diversity of synthetic molecules and the study of new bronchodilators, this work aimed at the synthesis and characterization of a thiophenic chalcone (E)-1-(benzo[d][1,3]dioxol-5-yl)-3-(thiophen-2-yl)prop-2-en-1-one (GA-01) and three unpublished derivatives, these being: 3-(benzo[d][1,3]dioxol-5-yl)-5- (thiophen-2-yl)-4,5-dihydro-1H-pyrazol-1-carbothioamide (GA-04), 3-(benzo[d][1,3] dioxol-5-yl)-1-phenyl-5-(thiophen-2-yl)-4,5-dihydro-1H-pyrazole (GA-05) and 1-(3- (benzo[d][1,3]dioxol-5-yl)-5-(thiophen-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)ethan-1-one (GA-06). Reaction yields varied between 93.8% and 31.83% and the compounds were obtained by precipitation, avoiding costs with exhaustive steps of purification. All molecules were subjected to in silico tests for their bronchodilator activity and for it’s pharmacokinetic properties. In the tests mentioned above, the substances GA-01, GA-05 and GA-06 showed potential bronchodilator activity.
  • ROXANE CARVALHO LIMA
  • “AVALIAÇÃO DOS EFEITOS DA ASSOCIAÇÃO ENTRE NITRATO DE SÓDIO E Lactobacillus plantarum WJL PARA O TRATAMENTO DA ATEROSCLEROSE EM CAMUNDONGOS FÊMEA KNOCKOUT PARA APOLIPOPROTEÍNA E”
  • Fecha: 05-jun-2020
  • Hora: 14:00
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  • A aterosclerose é uma patologia que está associada ao aumento dos níveis de colesterol e é considerada uma das principais causas que levam ao desenvolvimento de doenças cardiovasculares. Sua patogênese se caracteriza pela formação de placas (ateromas) ao longo das artérias, estimulada por um processo inflamatório. A inflamação ocorre devido aos danos às paredes arteriais provocados por diversos fatores de risco como diabetes, hipertensão e hipercolesterolemia. O nitrato inorgânico está amplamente disponível na dieta, podendo ser encontrado em vegetais de folhas verdes e beterraba. Na via nitrato-nitrito-NO, o nitrato é reduzido a nitrito por bactérias presentes na cavidade oral. Quando chega ao estômago, o nitrito é reduzido a NO. O óxido nítrico é importante para a homeostase vascular, uma vez que induz o relaxamento dos vasos, evitando assim a adesão de elementos celulares e diminuindo a inflamação. Por meio dessa ação, o óxido nítrico atua como antiaterogênico, sendo fundamental na gênese da aterosclerose. Probióticos são organismos vivos que trazem benefícios à saúde de seu hospedeiro e estão presentes no intestino de pessoas saudáveis. Estudos recentes demonstram sua possível capacidade de prevenir a aterosclerose por meio da redução do colesterol total, LDL e triglicerídeos. O presente estudo avaliou os efeitos da associação entre nitrato inorgânico e probiótico administrado por via oral para o tratamento da aterosclerose em camundongos fêmea knockout para apolipoproteína E. Foram utilizados camundongos das linhagens C57BL/6 e apoE-/-, onde os últimos receberam dieta aterogênica a partir de 8 semanas de idade. Os tratamentos duraram 12 semanas e consistiam em nitrato inorgânico (NaNO3) diluído em água (10mM) e probiótico (Lactobacillus plantarum WJL) administrado por meio de gavagem (109 UFC/mL). Dessa forma, os animais foram divididos em cinco grupos: C57BL/6, knockout para apolipoproteína E controle (apoE-/- CT), knockout para apolipoproteína E tratado com nitrato inorgânico (apoE-/- NT), knockout para apolipoproteína E tratado com probiótico (apoE-/- PB) e knockout para apolipoproteína E tratado com nitrato inorgânico e probiótico (apoE-/- NT+PB). O soro foi utilizado para a dosagem de colesterol total. As aortas foram coletadas para a avaliação de deposição de placas ateroscleróticas. Para a realização do teste de tolerância à glicose (GTT) e teste de tolerância à insulina (ITT) coletou-se uma gota de sangue da ponta da cauda dos animais através de uma incisão. Os camundongos apoE-/- CT (n = 7) mostraram incremento de 9 vezes nos níveis de colesterol total comparados com os C57BL/6 (n = 9) (816 ± 3.1 mg/dL vs. 90 ± 7.2 mg/dL) e o nitrato apresentou efeito hipocolesterolêmico (816 ± 3.1 mg/dL vs. 619 ± 92 mg/dL, (n = 4)). Além disso, a associação de nitrato + probiótico (n = 7) foi capaz de reduzir a deposição das placas ateroscleróticas. Os grupos C57BL/6 e apoE-/- CT mostraram perfis glicêmicos semelhantes, mas os grupos tratados com probiótico ou com a associação apresentaram pico aos 15 minutos da curva do teste de tolerância à glicose, retornando aos valores basais ao longo do tempo. Entretanto, nenhum dos tratamentos alterou o teste de tolerância à insulina. Assim, conclui-se que o tratamento com nitrato é suficiente para reduzir o colesterol na aterosclerose, entretanto a associação de nitrato e probiótico é necessária para que haja uma eficiente redução das placas ateroscleróticas.
  • IVYNNA SUELLEN JUSTINO VIDAL
  • Atherosclerosis treatment in apolipoprotein E knockout mice: association of a nitric oxide donor and probiotics
  • Fecha: 21-feb-2020
  • Hora: 09:00
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  • Atherosclerosis is a progressive vascular inflammation, initiated by the retention of low density lipoprotein (LDL), leading to oxidative stress, decrease nitric oxide (NO) bioavailability and vasodilation, increased vasoconstriction, and arterial obstruction due to the formation of atheroma plaques. Inorganic nitrates and probiotic microorganisms, from exogenous sources, have emerged as alternatives for the formation of NO, reduction in cholesterol levels, reduction of oxidative stress and reduction of atheroma plaques. Thus, pharmacological agents that are capable of improving vascular function are considered promising for combating atherosclerosis. The aim of the study was to evaluate the effects of the association of sodium nitrate (NaNO3) with Lactobacillus plantarum WJL on experimental atherosclerosis. Were used mice, male and female, from C57BL/6 and apoE-/-knockout strains for apolipoprotein E. The mice were divided into five experimental groups: c57 control (C), apoE-/- control (C), apoE-/- Nitrate (N), apoE-/- Probiotics (P) and apoE-/- association (P+ N). At the end of the treatment, the animals were euthanized, the blood was collected to perform the total cholesterol test, and the aortic artery was collected for the study of vascular function and histological analysis. Vascular function was assessed by constructing concentration-response curves to acetylcholine (Ach) and also concentration-response curves to PHE. To evaluate the direct response to smooth muscle, concentration-response curves to PHE and relaxation with sodium nitroprusside (SNP) were performed. The apoE-/- C (male: 769±11; female: 816 ± 3.1) animals showed an increase of approximately nine times in cholesterol levels in relation to C57 C (male: 85±2.1; female: 76±26). The apoE-/- N (male: 679±41; fêmeas:619±92) animals showed a reduction in cholesterol levels when compared to apoE-/-C. In atherosclerotic lesions, a reduction in the formation of plaques was identified in apoE-/- P males (1.9±1.3 vs. apoE-/- C: 12±0.97).The apoE-/- C animals (males: Rmax: 91.0 ± 3.69 and EC50: 6.71 ± 0.25; females: Rmax: 83.3 ± 5.10 and EC50: 7.0 ± 0, 15) didn’t present endothelial dysfunction in relation to ACh relaxation when compared to C57 C (males Rmax: 88.8 ± 5.54 and EC50: 7.11 ± 0.12; females: Rmax: 73.2 ± 4.74 and EC50: 7.1 ± 0.27) The treated animals also didn’t present significant differences, apoE-/- P (males: Rmax: 90.2 ± 18.9 and EC50: 7.2 ± 0.24; females: Rmax : 85.9 ± 2.50 and EC50: 7.2 ± 0.29), apoE-/- N (males: Rmax: 99.5 ± 6.65 and EC50: 7.0 ± 0.12; females: Rmax: 84.8 ± 10.5 and EC50: 6.4 ± 0.22), apoE-/- P + N (males: Rmax: 81.1 ± 5.35 and EC50: 7.0 ± 0.10 ; females: Rmax: 83.6 ± 5.22 and EC50: 6.9 ± 0.23. In the PHE response curves, apoE-/- C (males: Rmax: 102 ± 1.41 and EC50: 7.3 ± 0.079; females: Rmax: 101 ± 6.10 and EC50: 6.6 ± 0.31) showed marked endothelial dysfunction with high contractile response when compared to C57 C (males: Rmax: 59.1 ± 5.39 and EC50: 7.0 ± 0.066; females: Rmax: 72.0 ± 10.3 and EC50: 7.2 ± 0.18). In males the dysfunction of the apoE-/- N animals (Rmax: 46.67 ± 4.27 and EC50: 6.7 ± 0.18) and apoE-/- P + N (Rmax: 62.2 ± 7.91 e EC50: 6.9 ± 0.048), were reversed by the treatments. In the absence of endothelium, we observed a reduction in apoE-/- N (Rmax: 90.0 ± 5.66 EC50: 7.3 ± 0.097) and apoE-/- P + N (Rmax: 105 ± 6.98 and EC50: 7.4 ± 0.04) compared to apoE-/- C (Rmax: 145 ± 8.86 and EC50: 7.7 ± 0.06). In females facing the PHE curve with functional endothelium, endothelial dysfunction was reversed in the apoE-/- P groups (Rmax: 67.2 ± 5.41 and EC50: 7.0 ± 0.13) and apoE-/- P + N (Rmax: 78.6 ± 5.05 and EC50: 7.2 ± 0.073) in relation to apoE-/- C (Rmax: 101 ± 6.10 and EC50: 6.6 ± 0.31). In the absence of endothelium, we observed an increase in sensitivity in the apoE-/- N groups (Rmax: 112 ± 7.46 and EC50: 7.9 ± 0.06) and apoE- /- P + N (Rmax: 104 ± 5.57 and EC50: 7.6 ± 0.07) compared to apoE-/- C (Rmax: 117 ± 13.3 and EC50: 6.5 ± 0.28). In response to NPS, we observed a lower sensitivity of vascular smooth muscle in apoE-/- N males (Rmax: 128 ± 12.0 and EC50: 7.6 ± 0.12 vs. apoE-/- C Rmax: 115 ± 7.5 and EC50: 8.3 ± 0.02) and apoE-/- P + N (Rmax: 138 ± 7.3 and EC50: 7.4 ± 0.08) While there were no significant differences in females, between controls C57 C (Rmax: 126 ± 5.5 and EC50: 7.4 ± 0.27) and apoE-/- C (Rmax: 133 ± 11 and EC50: 8.1 ± 0.43) and among the treated groups apoE-/- P (Rmax: 110 ± 3.5 and EC50: 7.6 ± 0.32), apoE-/- N (Rmax: 122 ± 12 and EC50: 7.2 ± 0.04) and apoE-/- N + P (Rmax: 108 ± 1.1 and EC50: 7.2 ± 0.11) in relation to apoE-/- C. It is possible to conclude that the association of NaNO3 and L. plantarum WJL reversed endothelial dysfunction in experimental atherosclerosis in both sexes.
2019
Descripción
  • GEISI MARIA HENRIQUE DA SILVA
  • BIOPROSPECTION OF FILAMENTOUS FUNGI FOR PROTEASE PRODUCTION USING SKIMMED MILK AS A SUBSTRATE
  • Fecha: 30-nov-2019
  • Hora: 14:00
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  • Filamentous fungi are highly relevant organisms in the production of bioactives. An example of bioactive compounds are the proteases, enzymes involved in important metabolic changes, and increasingly used in industrial processes of several areas. However, it is necessary to produce enzymes with more attractive chemical characteristics for the industry, and with low productive cost. In view of this, the present work aimed the bioprospecting of filamentous fungi, for the production of protease by submerged fermentation, using skimmed milk like inductor substrate. Two fungal isolates from zebrafish eggs (Danio rerio), were subjected the macroscopic analysis (growth in Petri dishes) and microscopic analysis (coverslip culture). After, was analyzed the potential for proteolytic production in solid agar-milk medium, which showed that the genera Penicillium sp. and Clonostachys sp. has a positive protease activity. The isolates of Beauveria brongniartii and Paecilomyces sp., were previously analyzed.For the production of protease, 0.5 ml of conidia suspension (1x109 conidia / ml) was inoculated in 50 ml of skim milk (5%), for 5 days at 180 rpm. The proteolytic activity was determined using the azocazein and tricloacetic acid method (TCA). The crude extracts from the fermentation processes showed activities of 1.3 U / mL for Clonostachys sp., 4.6 U / mL for Penicillium sp., 6.9 U / mL for Beauveria brongniartii and 30.3 U / mL for Paecilomyces sp. The crude extract that demonstrated greater activity was submitted to partial purification using ethanol (0-80%). Where a significant increase has not been obtained in purity, the fraction purified by organic solvent with the highest total enzymatic activity (5.688 U) was submitted to purification by Ion Exchange Chromatography on a DEAE-Cellulose column, with a flow of 1mL / min and a mobile phase of 0.1M Tris-HCl pH 8, 0. The purified enzyme showed 35% of yield and a purification rate of 16.08 times. Thereafter, the protease from Paecilomyces sp. was characterized in terms of class, optimal temperature, thermostability and enzymatic kinetics. Serine protease inhibitors (SPIs) were used, but no inhibition was detected, demonstrating that the protease produced does not belong to this class. The enzymatic activity was analyzed at different temperatures (25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C and 60°C), showing greater activity at 50°C, for the crude extract and 35°C, for the purified enzyme. About the thermal stability, after the crude extract protease has been incubated at temperatures 25°C; 35°C; 40°C; and 45°C, was view greater stability in the first 2h, at 45°C. The kinetic parameters were founded experimentally, using different concentrations of azocasein - 0.1 to 3.0% (p / v) - resulting in the values of Km = 0.142% and Vmax = 6.8 U / mL. In view of the results obtained, we concluded that the fungi used were capables to produce protease by submerged fermentation, and that the skim milk medium is a potential substrate for the production of proteolytic enzymes.
  • YASMIM ESTRELA BATISTA GRISI
  • Proliferative potential of acetylcholine in cardiomyocytes
  • Fecha: 30-nov-2019
  • Hora: 08:00
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  • Cardiovascular diseases continue to be the biggest causes of death in the world for more than a decade, and Acute Myocardial Infarction (AMI) is the most frequently caused by diseases in Brazil. The damage caused by MI is caused by loss of functionality, local inflammatory process, ventricular remodeling, cystic fibrosis, which can culminate in heart failure until death. Strategies to revert this situation are studied and present several lines, and the most discrepant point indicates to be a cardiac regeneration. Acetylcholine (ACh) was subjected to a molecule with cardioprotective, anti-inflammatory effects and that cardiomyocytes have independent machines for the production of acetylcholine, was the main objective of studying whether an ACh has a proliferative role in cardiomyocytes after treatment with pyridostigmine. Observe the behavior in cardiomyocyte cultures of newborn rats from 3 to 5 days, when used with pyridostigmine (PIR), which is an inhibitor of acetylcholinesterase (enzyme that degraded ACh) and keep it modulated in these genes participate in the cell cycle. The chosen genes were Aurora BK, CDK1 and CCND2, data that were analyzed after an RT-PCR and compared with a B-ACTINA normalizer. It was concluded that the results show an ACh proliferative potential when PIR, in 0.1mM and 1mM, in the cardiomyocytes of neonatal rats, modulation of some genes in the cell cycle and potential in the application of treatment in AMI in neonatal rats.
  • BRENNDA MARTINS GABÍNIO
  • Anticanceriginal Evaluation of Nanoemulsions Containing R-(-)-Carvone
  • Fecha: 29-nov-2019
  • Hora: 08:30
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  • Characterized by a hyperproliferative disorder, cancer remains one of the leading causes of death worldwide. Where, breast cancer proves to be the most common and lethal among the female population in which even after treatment, many patients have deficiencies in physical and cognitive functioning, side effects that are often related to formulations administered during treatment. Showing that current therapies combine treatment with a high degree of uncertainty. Carvone is a monoterpene naturally found in several essential oils that in recent studies caused induction of apoptosis, inhibition of migration and arrest of the cell cycle in tumor cells presenting little or no toxicity in normal cells, however, it is characterized as volatile, presenting low solubility in water and inadequate biocompatibility in the form of native oil, therefore needing a transporter to efficiently reach cancer cells. Among the delivery systems, there are nanoemulsions that are emulsions of submicron size and systems used for systemic delivery of biologically active agents. Thus, the present study aimed to evaluate the in vitro anti-cancer potential of R - (-) - free and nanoencapsulated carvone on breast cancer cell culture. The nanoemulsions were produced in concentrations of 5mg / mL, 12.5 mg / mL, 25 mg / mL, 37.5 mg / mL and 50 mg / mL by the ultrasonic emulsification method and characterized by of their physical-chemical aspects, determining the particle size, polydispersity index and zeta potential, in addition to obtaining the calibration curve and the pH and conductivity values. In vitro cytotoxicity tests were also performed on the fibroblast (L929) and mammary gland (4T1) lines, measuring cell viability using the MTT reduction test and we performed the wound healing test observed at 0h, 24h and 72h. The results found show nanoparticles with average size varying between 133nm and 183nm, not proportional to the amount of drug used, average zeta potential of -36.4mV and polydispersion index less than 0.240. Vehicle toxicity was the next control group and showed values above 100%, inducing possible proliferation. The emulsions analyzed were cytotoxic for the breast cancer lineage, where the nanosystem shows good activity and potential for potentiation of cytotoxic effects when compared with a free R-CV. The results shown a fast migration in the control group, with the permanence of the migration potential with the free drug and alteration in the adhesion of the matrix cell in cancer cells. Therefore, we achieved satisfactory formulation with a significant anticancer effect on breast cancer cells in a short period of time, showing represents a promising formulation to assist the conventional treatments used for this disease.
  • BRUNO HANRRY MELO DE OLIVEIRA
  • SYNTHESIS AND BIOLOGICAL EVALUATION OF (±) 4-O-DEMETILKADSURENIN M, AND ITS ANALOGS
  • Fecha: 27-nov-2019
  • Hora: 08:00
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  • An important class of biologically active natural molecules are lignoids, molecules formed exclusively by the coupling of phenylpropanic groups (C6-C3) n, where 'n' is restricted to a few units, 1,2,3 etc. Examples of lignoids include lignans, neolignans, allolignans, oligolignoids and heterolignoids. This paper presents an approach to neolignans that together with lignans represent 90% of the lignoids described in the literature. Neolignans have a huge structural diversity, nature itself offers us a very wide variety of these compounds, besides, from a chemical point of view, and several neolignans have already been used as starting material for the synthesis of derivatives of natural products with various purposes, such as optimizing product activity and studying the structure-activity relationship. These molecules also have numerous pharmacological activities described in the literature, such as antitumor, antibacterial, antimicobacterial, trypanosomicide, schistosomicide, leishimanicide, etc. activities. There are several ways to synthesize these types of compounds in the laboratory, such as oxidative couplings obtained through FeCl3, Ag2O, peroxities, among others. Thus, the objective of this study was to synthesize neolignan substances for the biological evaluation of them in human tumor cells HL-60 (acute promyelocytic leukemia), HCT-116 (human colorectal carcinoma), MCF-7 (human mammary adenocarcinoma). , K562 (chronic myeloid leukemia) and non-tumor lineage L929 (murine fibroblast), as well as bacterial strains Escherichia Coli UFPEDA224, Staphylococcus aureus UFPEDA02 and Pseudomonas aeruginosa UFPEDA416.
  • CAMILA DE CASTRO BARBOSA
  • Evaluation of effects induced by acetate nitrate of 4 - Nitrooxybutil ( NHPR1) on the cardiovascular system of normotense rats
  • Fecha: 26-nov-2019
  • Hora: 09:00
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  • Systemic arterial hypertension (SAH) is related to endothelial dysfunction attributed mainly by the reduction of nitric oxide (NO) bioavailability in the vascular wall and increased oxidative stress, resulting in chronic and abnormal increase in peripheral vascular resistance. Thus, it is characterized by high blood pressure levels, resulting in a change in vascular tone, where NO is the main endothelium-derived relaxing factor (EDRF). Nitric oxide plays a key role in the control and regulation of blood pressure through its influence on peripheral vascular resistance and vascular tone. Problems related to NO bioavailability are prime factors for the development, progression and maintenance of hypertension. The use of compounds that increase the bioavailability of NO has been a therapeutic strategy for the treatment of cardiovascular disorders for decades. Thus, the objective of the present study is to evaluate the effect of 4 - nitrooxybutyl acetate nitrate (NHPR1) on the cardiovascular system of normotensive rats. From the biological prediction, NHPR1 showed a vasodilating effect where the NO donor mechanism (Pa 0.849) was identified using the cutoff line Pa> 0.7 Pi = 0.000. In ex vivo tests, in isolated cranial mesenteric artery rings pre-contracted with FEN (1μΜ), NHPR1 was able to induce vasorelaxant effect in both functional endothelial rings (Emax = 100.0 ± 6.116, n = 10). in rings where the endothelium was removed (Emax = 100.0 ± 11.47, n = 7). After an electromechanical contraction with KCl (60 mM), NHPR1 showed no altered vasorelaxant response (Emax = 132.7 ± 7.103, n = 7). To assess the participation of the NO / GCs pathway was used ODQ (10 μM), a selective inhibitor of the soluble guanylate cyclase enzyme, which attenuated the vasorelaxant response to NHPR1 (Emax = 99.24 ± 7.338, n = 8). When performing non-selective blockade of potassium channels with ASD (3 mM), the vasorelaxant response of NHPR1 was not altered (Emax = 118.1 ± 4.828, n = 13). In the tolerance test, nitrate showed alteration in its induced response, which observed the occurrence of shift of the curve to the right, suggesting that there is the development of tolerance in the vasorelaxant response of NHPR1. In the evaluation of toxicity, oral administration of NHPR1 (300 and 2000 mg / kg) did not promote changes in body weight and organ weight, as well as food and water consumption compared to the control group. Acute administration of NHPR1 (1, 5, 10, 20, 50 mg / kg, iv) induced hypotension in normotensive animals (-4.06 ± 1.01; -6.18 ± 1.25; -15.39 ± 0.65, -11.71, ± 3.04, -17.19 ± 1.99 mmHg, respectively) and bradycardia (6.48 ± 3.64; 62.61 ± 12.88; 48.21 ± 9 , 08; 76.31 ± 13.44; 103.75 ± 12.13 bmp) in a dose - dependent manner. Thus, the vasorelaxant response promoted by NHPR1 possibly occurred due to its vasodilating effect involving NO release and subsequent activation of the NO-GCs-PKG pathway. This mechanism of action may be contributing to the hypotension and bradycardia observed in normotensive animals.
  • ADRIELLY SILVA ALBUQUERQUE DE ANDRADE
  • Baculovirus: In Vitro Production of Anticarsia Bioinsecticide and Construction of Rabies lyssavirus Glycoprotein (GPV) Expression Vector
  • Fecha: 31-may-2019
  • Hora: 10:00
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  • Baculoviruses are excellents agents for controlling the insect population in the wild and therefore have been successfully used as viral bioinsecticides in agriculture. Moreover, thanks to the knowledge of the in vitro infection process in host cells, baculoviruses are also used as recombinant protein expression vectors. Thus, the present study aimed to produce in vitro wild and recombinant baculovirus in insect cells for future application as viral bioinsecticide and expression of recombinant proteins. For this, the IPLB-SF21 insect cells, adapted to the suspension culture, were used in the first (P1), second (P2) and third pass (P3) Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) extracellular virus infection processes. The variables analyzed were cell growth and viability, substrate consumption and viral productivity of the different passages of the viral inoculum in the infection processes. The data obtained were used to determine the kinetic parameters: Specific growth rate (μx); Substrate consumption (qS) and; Production of occlusion bodies (qOB). As a result, the control cells had a μx equal to 0,570 d-1 and the infected cells presented infection rates 0,204 d-1 (P1), 0,131 d-1 (P2) and 0,335 d-1 (P3), indicating that baculovirus reduced the growth of infected cells when compared to control cells. As for qOB, 0,93 OB(mL.d-1), 0,70 OB(mL.d-1) and 1,34 OB(mL.d-1) were obtained for P1, P2 and P3, respectively, and it can be seen that it is directly proportional to the concentration of infected viable cells. Based on these velocities, it was possible to calculate the yield of polyhedra per milligram of glucose, 1,18x106 OB.mg-1, 0,85x106 OB.mg-1 and 2,55x106 OB.mg-1, demonstrating the importance of the kinetic parameters when comparing productivity between the systems and ensuring a better yield of the viral bioinsecticide. For expression vector construction, the total RNA obtained from brain tissue from mice infected with inactivated R. lyssavirus was used for cDNA construction and cloning into competent E. coli cells. Plasmid DNA was extracted, but there was a low amplification of the glycoprotein gene.
  • ANAUARA LIMA E SILVA
  • Leaf morphoanatomy and Fourier transform infrared spectrophotometry (FTIR) as support for taxonomy and quality control of ethnodrugs of Solanum species sections Erythrotrichum Child, Micracantha Dunal and Polytrichum Child (Solanaceae), pro parte
  • Fecha: 16-may-2019
  • Hora: 14:00
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  • A morphoanatomic study and a Fourier-transform infrared spectroscopy (FTIR) and multivariate analysis were performed on leaves of eleven species of Solanum subg. Leptostemonum, belonging to the sections Erytrhrotrichum, Micracantha and Polytrichum, in order to find additional support to the taxonomy of these sections and also to the quality control of their ethnodrugs. Samples of fresh and dry specimens were used for the morphological studies. Dissociations of fragments of leaf blades were performed for the epidermis analysis, following the usual protocol in anatomy, in addition to scanning electron microscopy. For the mesophyll, midrib and petiole were performed cross sections, performed by free hand, stained with a solution of astra blue and safranin, analyzed and photographed under the light microscopy. A hierarchical grouping analysis was performed with a matrix of 61 characters. For IR spectrophotometry, the samples were prepared directly by pressing on a KBr pellets, placed and analyzed on the support on the Cary 630 FTIR Spectrometer. As results, it was observed that some characters were common to the studied species: leaves with discolored blades, epidermis with sinuous anticlinal walls, dorsiventral mesophyll, biconvex midrib with a central bicollateral bundle. However, some characters were distinctive for each section: in Erythrotrichum, predominated the elliptic to oval-elliptic leaf blade, the petioles are cylindrical and canaliculate, semicircular to circular with lateral projections; in Micracantha, acute-elliptic to lanceolate-elliptic leaf blades, canaliculate petiole with oval, circular or semicircular contour; in Polytrichum the oval-elliptic to oval-cordate leaf blade, cylindrical or winged petiole, circular or semicircular with or without lateral projections. The phenogram generated from the morpho-anatomical characters have nine artificial clusters and reunited the species into two large groups: Erythrotrichum-Micracantha and Polytrichum. The FTIR spectra of visual inspection sampling were similar. Eight species showed peaks suggestive for phenolic esters and / or methyl esters and only one species for amides. The study revealed the importance of FTIR spectroscopy to distinguish complex groups of Solanum species, as well as highlighting relevant characters that can be used as an additional tool to the sectional and interspecific delimitations of the species studied, especially indument, epidermal morphology and trichomes, leaf margins and petioles, which constitute a set of characters that support the taxonomy of the sections studied and Solanum subg. Leptostemonum, and also contribute to the quality control of their ethnodrugs.
  • UBIRATAN RIBEIRO DA SILVA FILHO
  • Production, extraction, purification and potential antimicrobial activity of the biosurfactant produced by Bacillus safensis MF326565 isolated from sugarcane soil
  • Fecha: 30-abr-2019
  • Hora: 14:00
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  • Surfactants are amphipathic molecules capable of interacting with surfaces of different polarities and because they are derived from petroleum, they have limitations and problems associated with their use, among which environmental ones stand out. Biosurfactants are an alternative because they demonstrate environmentally desirable characteristics that depend on the metabolism of organisms, associated with technologies that favor their production. In this context, we highlight the use of sugarcane molasses as an alternative source of substrate. Therefore, the objective of this work was to produce biosurfactant in bench top bioreactor using Bacillus safensis, MF326565, isolated from sugarcane soil and sugarcane molasses as an alternative substrate. The experiments were carried out preliminarily in shaker (1% of sugarcane molasses and 1% of yeast extract - w / v). The diagnosis of biosurfactant production was made by growth analysis, surface tension and emulsification index. In bioreactor, they followed the same analyzes, besides the characterization by FTIR and evaluation of antimicrobial activity. The results of shaker cultivation were quite satisfactory regarding growth and biosurfactant production, since the microorganism was able to use the substrate and produce biosurfactant, which presented emulsification index higher than 50% in motor oil, besides presenting reduction. surface tension by 32%. In bioreactor a growth of approximately 3.0 g / l of biomass and 1.5 g / l of biosurfactant was observed and with much higher emulsification in motor oil, vegetable oil and kerosene. The biosurfactant showed stability at temperature (15 ~ 60 ° C), pH (2.0 ~ 8.0), salinity (6 ~ 10% NaCl), and emulsification remained stable for more than 15 days. Infrared spectrograms (FTIR) of the purified product from the bioreactor experiments showed similarity with the surfactin standard, showing its lipopeptide nature. The biosurfactant showed antimicrobial activity against S. aureus, B. cereus, B. safensis, B. subtilis, C. albicans and Acinetobacter sp., bactericidal activity at concentrations of 12.5 mg / mL for Acinetobacter sp. and S. aureus and concentrations below 3.0 mg / mL for Bacillus used in this study. Thus, it is concluded that Bacillus safensis, MF326565, was able to produce biosurfactant from an alternative substrate (molasses from sugarcane) and that it was effective in emulsifying hydrophobic compounds, besides presenting biological activity.
2018
Descripción
  • RAFAEL LIMONGI DE SOUZA
  • INVESTIGATION OF THE CORRELATION BETWEEN THE POTENTIAL OF BIOREMEDIATION AND THE PRODUCTION OF OXIDOREDUTASES BY Pseudomonas aeruginosa
  • Fecha: 30-nov-2018
  • Hora: 14:00
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  • Environments contaminated by polycyclic aromatic hydrocarbons are challenges that can be minimized with the use of microorganisms with degradation potential. The objective of this work was to evaluate the degradation capacity of anthracene and pyrene by a Pseudomonas aeruginosa isolate and to correlate with laccase and manganese peroxidase activity. Three isolates of Pseudomonas aeruginosa (TGC-02, 03 and 07) and one positive control with Aspergillus niger were evaluated. The experiments were divided into two steps: screening with P. aeruginosa isolates was initially screened for emulsification and drop collapse tests. The higher hydrocarboclastic activity was observed for the TGC-02 isolate, which degraded more than 30% of the contaminants tested. Subsequently, the biodegradation assays of the anthracene and pyrene contaminants by the selected TGC-02 isolate and compared to an A. niger isolate were conducted. The assays lasted 25 days at room temperature in microcosms containing 30 mL of distilled water, contaminated with 50 mg / L of pyrene or anthracene, under static incubation. For each point of analysis, pH, biomass, cell viability, total proteins and enzymatic activity were measured. The content of the contaminants was determined by gas chromatography coupled to flame spectrometry after 25 days of the bioprocess. No specific activity was observed for laccase or manganese peroxidase in the tested isolates, and it was not possible to correlate the degradation capacity with the specific activity of these enzymes. However, when analyzing the total protein results, it was observed that during the 25 days there was an increase in the concentrations of total proteins in presence of the contaminants, especially for P. aeruginosa TGC-02. The TGC-02 and A. niger isolates cultured in the presence of the contaminants presented viability during the 20 days of the bioprocess. Discounting the abiotic losses, the positive control A. niger degraded 28.11% and 30.70% of the pyrene and anthracene, respectively. P. aeruginosa TGC-02 presented better results, reducing pyrene and anthracene in 37.83% and 33.01%, respectively. A. niger is a fungus known as a degradation agent of aromatic compounds, however, in this work P. aeruginosa proved to be more efficient in the degradation of pyrene and anthracene in the liquid medium in 25 days. The results demonstrate the potential of bioremediation of P. aeruginosa for the removal of pyrene and anthracene, apparently independent of the production of manganese peroxidase and laccase.
  • JOSEANNE DANIELE CEZAR RIBEIRO
  • "Efeitos da Angiotesina-(1-7) na toxicidade induzida por tratamento com Doxorrubicina"
  • Fecha: 23-nov-2018
  • Hora: 14:00
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  • Conforme dados do Instituto Nacional do Cancer Jose Alencar Gomes da Silva (INCA), no bienio 2018-2019, para cada ano, aproxima-se um valor de 600 mil novos casos de cancer no Brasil. Como saida terapeutica para atingir o importante numero populacional acometido por essa doenca, existem os antibioticos antitumorais, agentes antineoplasicos onde esta inserida a doxorrubicina. Apesar de sua eficacia no combate as celulas tumorais, sua utilizacao pode provocar reacoes adversas como a cardiotoxicidade. Nas abordagens farmacologicas uma substancia que tem potencial no quesito protecao cardiaca e a Angiotensina-(1-7), um heptapeptideo endogeno participante do sistema renina-angiotensina. Diante disso, o objetivo deste estudo foi investigar os efeitos da Angiotensina-(1-7) na prevencao e tratamento da toxicidade provocada pela Doxorrubicina. Neste estudo foram utilizados 35 ratos Wistar adultos (Rattus novergicus), pesando de 160-278g, provenientes do Bioterio Prof. Thomas George da Universidade Federal da Paraiba. Os animais passaram por um ciclo de seis semanas de experimentacao, sendo divididos em 6 grupos: Controle Solucao salina, Controle Ang-(1-7), Controle doxorrubicina, Preventivo 1 e 2, e grupo Tratamento. Os animais foram submetidos a 3 avaliacoes ecocardiograficas ao longo dessas semanas e ao final do tratamento foram eutanasiados, com posterior avaliacao do peso dos seguintes orgaos: coracao, pulmao, rins (D e E), baco e figado, alem da analise do aspecto morfologico macroscopico do organismo dos animais (com registro fotografico). Na avaliacao da sobrevivencia os grupos Controle Solucao salina e Controle Ang-(1-7) chegaram ao final do experimento com todos os animais vivos, diferente dos grupos de tratamento que receberam Ang-(1-7) e doxorrubicina. Nos grupos controle Doxo e Tratamento houve uma queda no valor da massa corporal dos animais, alem de caracteristicas como apatia, desidratacao e fezes diarreicas com muco. O grupo Controle Solucao Salina na avaliacao da necropsia apresentou uma morfologia com focos hemorragicos nos pulmoes, manchas brancas em ambos os rins e apenas em um dos ratos desse grupo foi capaz de observar a normalidade morfologica macroscopica. O grupo Controle Ang-(1-7) apresentou caracteristicas morfologicas normais. Os animais dos grupos que receberam a doxorrubicina apresentaram rigidez caracteristica nos orgaos, em especial no figado, alem do acumulo de liquido na cavidade peritoneal e focos hemorragicos em pulmoes. Em relacao a funcao ventricular esquerda os animais demonstraram um perfil semelhante entre os grupos de tratamento comparado ao grupo Controle. No que compete a avaliacao da massa dos orgaos dos animais analisados, o grupo Controle Ang-(1-7) se comportou igual ao grupo Controle Solucao salina. A massa cardiaca isolada apresentou reducao nos grupos Controle doxorrubicina, Preventivo 1 e Tratamento, mas em relacao a massa corporal, verificou-se que a proporcao nao foi alterada entre os grupos. A massa pulmonar isolada mostrou-se alterada nos grupos Preventivo 1 e Preventivo 2 e na relacao massa do orgao/massa corporal do animal esse aumento no volume pulmonar tambem pode ser observado nos grupos Controle doxorrubicina e Tratamento. O baco dos animais apresentou alteracao apenas no grupo Preventivo 2. Nao encontramos diferenca na massa dos rins (D e E) entre os grupos de tratamento, mas constatamos que se observado a relacao massa do orgao/massa corporal houve aumento nos grupos que receberam apenas a doxorrubicina no inicio do experimento. Por fim, a massa do figado dos animais tambem nao apresentou diferenca entre os grupos quando observado isoladamente, mas quando considerada a massa corporal do animal os grupos Controle doxorrubicina, Preventivo 1 e Tratamento manifestaram uma diferenca importante do grupo Controle Solucao salina. Diante dos resultados encontrados nao foi possivel analisar prevencao de cardiotoxicidade pela Ang-(1-7), e que quando administrada em associacao com a doxorrubicina os efeitos foram demasiadamente deleterios.
  • ÉSSIA DE ALMEIDA LIMA
  • Atividade anti-inflamatória da pterocarpa LQB 118 em macrófagos murinos
  • Fecha: 25-sep-2018
  • Hora: 10:00
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  • A LQB 118 e uma pterocarpanoquinona, molecula hibrida sintetica resultante da uniao de dois grupos de moleculas naturais bioativas, os pterocarpanos e as naftoquinonas. Esta molecula possui varias atividades biologicas relatadas, incluindo atividade anti-inflamatoria em modelo de inflamacao alergica pulmonar. No entanto, ainda nao ha relatos na literatura sobre a atividade da LQB 118 em outros modelos de inflamacao. Dessa forma, o objetivo deste trabalho foi avaliar, em macrofagos peritoneais murinos, o papel da pterocarpanoquinona LQB 118 no processo inflamatorio. Inicialmente, camundongos Swiss receberam uma injecao de tioglicolato (4%) via intraperitoneal (i.p.). Quatro dias depois, os macrofagos peritoneais foram obtidos e cultivados na concentracao de 2 x 105 cels/poco. As celulas foram tratadas com a LQB 118 (5 µM, 1 µM, 0.5 µM e 0.1 µM) na presenca ou ausencia do estimulo inflamatorio, zimosan (0,2 mg/mL), por 24 h. Em seguida, os sobrenadantes foram recolhidos para a quantificacao das citocinas (TNF-α, IL-1β e IL-6) por ELISA. A viabilidade celular foi determinada pelo metodo de MTT. Para analise do mecanismo de acao da molecula, os macrofagos foram cultivados na concentracao de 1,5 x 106 cels/poco e tratadas como ja descrito. Apos 24 h, as celulas foram utilizadas para analise da expressao de moleculas envolvidas na inflamacao por citometria de fluxo. Para isso, os macrofagos foram incubados com os anticorpos anti-TLR2, anti-CD69 e anti-P-p38. Como esperado, o zimosan induziu aumento da producao das citocinas pro-inflamatorias avaliadas. Por sua vez, o tratamento com a LQB 118 foi capaz de reduzir significativamente os niveis das citocinas TNF-α, IL-1β e IL-6 no sobrenadante da cultura celular. Alem disso, o tratamento com a LQB 118 foi capaz de reduzir a expressao da molecula CD69, assim como o aumento da fosforilacao da MAPK p38 induzida por zimosan. Adicionalmente, a LQB 118 foi capaz de modular negativamente a expressao do TLR2 na presenca do estimulo inflamatorio. Os resultados obtidos sao independentes de morte celular, visto que nenhuma das concentracoes de LQB 118 utilizadas interferiu na viabilidade dos macrofagos. Dessa forma, este trabalho demonstrou pela primeira vez o efeito anti-inflamatorio da LQB 118 em macrofagos peritoneais murinos.
  • ANNE KALIERY DE ABREU ALVES
  • AVALIAÇÃO DO EFEITO DA OUABAÍNA NA VIABILIDADE E MODULAÇÃO DE CITOCINAS EM TIMÓCITOS E LINFÓCITOS EXPOSTOS A RADIAÇÃO ULTRAVIOLETA
  • Fecha: 29-may-2018
  • Hora: 10:00
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  • A ouabaina e um glicosideo cardiaco que foi descoberto no plasma humano em 1991 (HAMLYN et al., 1991). Essa substancia endogena e capaz de inibir a Na+/K+-ATPase e vem sendo amplamente estudada por sua capacidade em interferir em varios mecanismos que regulam e mantem a homeostase. Portanto o objetivo desse trabalho foi avaliar in vitro e in vivo a acao da ouabaina na viabilidade celular e no perfil de citocinas pro-inflamatorias em celulas do timo e linfonodo mesenterico expostas a radiacao UV. Para isso foram usados camundongos Swiss albinus e realizada a cultura de celulas do timo e linfonodo mesenterico. Essas celulas foram tratadas com oubaina nas concentracoes de 100,10, 1 µM e 100 nM na placa de 96 pocos, como tambem atraves do pre-tratamento com ouabaina na dose de 0,56 mg/kg por tres dias atraves da injecao intra-peritoneal dessa dose. As celulas obtidas da maceracao dos orgaos foram centrifugadas, resuspensas e contadas. A concentracao foi ajustada para 4x105cels/ml com PBS e plaqueadas para a exposicao a UVC e UVB por 2 minutos. Apos o estimulo, adicionou-se o meio RPMI suplementado com 10% de SFB e manteve-se essas celulas em cultura por 6 e 24 horas. A viabilidade foi analisada com o MTT 5 mg/mL. O sobrenadante da cultura foi utilizado para determinar os niveis das citocinas proinflamatorias TNF-α e IL-6 pelo metodo de ELISA. Nossos resultados mostraram que o pretratamento com ouabaina na dose de 0,56 mg/Kg na cultura de 6 horas apresentou um efeito citoprotetor nos timocitos e linfocitos linfonodo mesenterico expostos a UVC e UVB. Esse mesmo tipo de celula em cultura por 6h e 24h na presenca de ouabaina em diferentes concentracoes 100,10, 1 µM e 100 nM, apresentou efeito em 24 horas mediante o estimulo da UVC, esse dado nao foi observado para UVB. A concentracao de 100 nM protegeu os timocitos e linfocitos do linfonodo mesenterico da morte celular. No sobrenadante dessa concentracao houve aumento do TNF-α e reducao de IL-6 quando, e no pre-tratamento com 0,56 mg/Kg nao houve alteracao dessas citocinas comparada ao grupo que recebeu radiacao. Esses resultados contribuem para o entendimento da sua acao ouabaina sobre parametros importantes do processo inflamatorio, viabilidade e citocinas, desencadeado por radiacao ultravioleta.
  • NATHALIA SOUZA BEZERRA
  • “Eficiência de fungos entomopatogênicos sobre formigas cortadeiras (Hymenoptera: Formicidade)”
  • Fecha: 27-feb-2018
  • Hora: 09:00
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  • do Neotropico. Atualmente, o principal metodo de controle empregado e o uso de inseticidas quimicos. Contudo, existe uma crescente preocupacao acerca dos efeitos negativos das aplicacoes frequentes de inseticidas em organismos nao-alvo, na saude humana e no ambiente. Os fungos entomopatogenicos podem, naturalmente, infectar insetos hospedeiros via penetracao direta da cuticula e tem sido estudados em todo o mundo como promissores agentes de controle biologico. Dito isso, o objetivo desse trabalho e avaliar a eficiencia de isolados de Beauveria bassiana, Beauveria brongniartii, Metarhizium anisopliae var. anisopliae, Paecilomyces sp. e Aspergillus sp. sobre formigas cortadeiras em laboratorio. Alem de analisar o efeito do Fipronil sobre o crescimento vegetativo e conidiogenese dos isolados fungicos. Para cada especie de fungo, foi realizado um experimento com 90 exemplares de formigas cortadeiras, correspondendo a tres repeticoes de 10 formigas para cada tratamento (Tratamento 1Grupo controle = agua destilada autoclavada + Tween 80 0,01 %; Tratamento 2 = suspensao de 1,0 x 104 conidios.mL-1 + Tween 80 0,01 %; Tratamento 3 = suspensao de 1,0 x 108 conidios.mL-1 + Tween 80 0,01 %). As formigas foram transferidas individualmente para placas de Petri contendo papel de filtro com 1 mL da suspensao de conidios e bolas de algodao umedecidas com uma solucao de mel a 10 %. As placas foram mantidas a temperatura de 25 ± 2 °C e umidade relativa de 70 ± 10 % e avaliadas a cada 24 horas durante 10 dias, para observacao da extrusao dos fungos e confirmacao da morte dos insetos pelos patogenos. Para determinar o efeito do inseticida sobre o crescimento vegetativo e conidiogenese dos isolados fungicos, 0,8 g/L do produto e 0,3 g/L de penicilina foram adicionados ao meio de cultura Agar-Sabouraud-Dextrose. Apos 10 dias, o diametro de cada colonia foi medido com o auxilio de uma regua e os conidios foram contados em camara de Neubauer. O Fipronil foi, entao, classificado quanto a toxicidade in vitro sobre os isolados. Como resultado do bioensaio, obteve-se que as duas concentracoes de conidios utilizadas (1,0 x 104 e 1,0 x 108 conidios.mL-1) foram capazes de assegurar a infeccao e o progresso da doenca nas formigas cortadeiras, com a mortalidade media das formigas variando entre 4,80 ± 2,10 e 9,99 ± 0,00 e o TL50 variando entre 6,01 e 10,22 dias. Em relacao as concentracoes de conidios, observou-se que, em alguns casos, o aumento da concentracao contribuiu para se elevar a mortalidade nos insetos. Entre todos os isolados, Aspergillus sp. foi o que apresentou as maiores mortalidades medias (9,99 ± 0,10 e 9,99 ± 0,00) e os menores TL50 (6,47 e 6,01 dias) para ambas as concentracoes de conidios. Os isolados fungicos tratados com Fipronil apresentaram uma reducao no crescimento vegetativo. Contudo, o inseticida nao comprometeu significativamente a conidiogenese dos mesmos. Conclui-se, portanto, que o bioensaio descrito no presente estudo permitiu a descoberta de isolados com grande potencial para serem avaliados em campo para o controle biologico de formigas cortadeiras, contribuindo para o desenvolvimento de alternativas mais eficazes e menos danosas ao ambiente para o controle desses insetos-praga e que o Fipronil, na concentracao testada, mostrou-se “compativel” para os isolados fungicos no teste de compatibilidade in vitro.
2017
Descripción
  • EMANUELE CARDOSO DIAS
  • PRODUÇÃO DE BIOSSURFACTANTES POR FERMENTAÇÃO SUBMERSA UTILIZANDO EXTRATO AQUOSO DA ALGAROBA {PROSOPIS JULIFLORA (Sw) DC} COMO SUBSTRATO
  • Fecha: 28-nov-2017
  • Hora: 14:00
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  • Biossurfactantes sao moleculas anfipaticas sintetizadas por micro-organismos que agem nas interfaces agua/oleo ou oleo/agua reduzindo as tensoes superficiais e interfaciais entre elas e assim tornando alguns compostos mais misciveis. Em comparacao aos surfactantes sinteticos que estao no mercado, apresentam diversas vantagens, como a biodegradabilidade e baixa toxicidade conferindo-lhes relevada importancia. A utilizacao de substratos nao convencionais, sobretudo renovaveis, e uma estrategia de producao de biossurfactantes devido ao baixo custo oferecido, constituindo um dos fatores mais importantes para a viabilizacao economica em escala industrial. Neste trabalho, a producao de biossurfactantes por Bacillus subtillis utilizando o extrato aquoso da algaroba como substrato alternativo foi estudada, utilizando-se um planejamento fatorial 2² com triplicata no ponto central, para avaliar a influencia da concentracao desse substrato e extrato de levedura nessa producao. Diante dos resultados, o aumento da concentracao do extrato aquoso de algaroba com suplementacao de extrato de levedura favoreceu o crescimento do micro-organismo e a producao de biossurfactantes, comprovados pelo planejamento fatorial realizado. O indicativo de producao de biossurfactante, indice de emulsificacao, foi maior no cultivo utilizando a suplementacao a 0,5 e 1,0 % de extrato de levedura. Com relacao ao indice de emulsificacao, foram alcancados niveis equivalentes aos encontrados na literatura e tambem se destacando, o produto oriundo do ensaio em biorreator que evidenciou uma eficacia proporcional ao surfactante sintetico SDS. E por fim, considerando a similaridade apresentada nos espectrogramas com os IR’s da surfactina industrial, conclui-se tambem que o biossurfactante encontrado neste processo fermentativo submerso correspondem a lipopeptideos. Os resultados indicam a viabilidade de producao de biossurfactantes a partir do extrato aquoso da algaroba, substrato de baixo custo e abundante na regiao Nordeste.
  • CICERO ANTHONYELSON TEIXEIRA DUNES
  • “Síntese de Derivados Dihidroquinazolinonas com Potenciais de Inibição para Ricina”.
  • Fecha: 13-nov-2017
  • Hora: 09:00
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  • A Ricinus communis L., comumente denominada de mamoneira, tem ganhado atencao da industria em funcao das propriedades dos seus derivados, a producao de oleo, principal produto da mamona, gera quantidades consideraveis de subprodutos que necessita de uma destinacao, mas que e inconveniente pela presenca da ricina, uma glicoproteina altamente toxica, sendo tambem a ricina apontada como potencial agente de bioterrorismo, faz-se necessario encontrar meios de neutraliza-la. O retro-2, com estrutura heterociclica, esta entre as moleculas descritas como capaz de bloquear eficientemente o transporte da ricina, com isso o objetivo desse trabalho foi sintetizar derivados dihidroquinazolinonas analogas ao retro-2, identifica-las e avaliar as suas atividades de citotoxicidade, inibicao para a RTA e binding com a holoricina. Foram sintetizados quatro moleculas a partir da condensacao da 2aminobenzanilida com aldeidos fenolicos, foram elas: 2-(4-hydroxy-3,5-dimethoxyphenyl)-3phenyl-2,3-dihydroquinazolin-4(1H)-one trivialmente chamada de C100, composto solido, com rendimento de reacao de 0,75 g (70,34%), com ponto de fusao de 212°C e Rf de 0,3 (em eluente de 6:4 de AcOEt/hexano); a 2-(3,4-dihydroxyphenyl)-3-phenyl-2,3dihydroquinazolin-4(1H)-one chamada de C200, um solido com rendimento de 0,4842 g (72,92%), com ponto de fusao de 195°C e Rf de 0,47 (em eluente de 6:4 de AcOEt /hexano); a 2-(4-hydroxy-3-methoxyphenyl)-3-phenyl-2,3-dihydroquinazolin-4(1H)-one chamada de C300, composto solido de cor branca, obtido a partir de uma reacao com rendimento de 59,57%, com ponto de fusao de 188°C e Rf de 0,67 (em eluente de 6:4 de AcOEt/hexano) e a 2-(2-hydroxyphenyl)-3-phenyl-2,3-dihydroquinazolin-4(1H)-one chamada de C400, composto de cor branca e estado solido, com rendimento de 48,69%, ponto de fusao de 183°C e Rf de 0,75 (eluente de 6:4 de AcOEt/hexano); todas se mostraram pouco toxicas em linhagens de celulas HUVEC e THP-1 a uma concentracao de ate 200 µM, calculado o IC50 apenas na THP-1 em 149 µM da C100, e 182,8 µM para a C200 na mesma linhagem. O teste de inibicao da RTA foi inconclusivo visto que o padrao positivo (retro-2) nao foi capaz de inibi-la, e o binding moleculas/ricina mostrou interacao entre elas.
2016
Descripción
  • ELTON JOSÉ FERREIRA CHAVES
  • Simulação Molecular de Inibidores da Subunidade da Ricina, RTA
  • Fecha: 29-nov-2016
  • Hora: 14:00
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  • A Ricinus communis, especificamente o seu fruto, a mamona, tem ganhado atencao da industria, midia e governos, devido as propriedades de seus derivados, especialmente a ricina e o oleo. Do total de acidos graxos extraidos da mamona, 90% compreende ao acido ricinoleico. Alem do oleo, os coprodutos gerados durante a producao tem ganhado bastante atencao, sendo os mais importantes, a casca do fruto e a torta. Tais coprodutos apresentam quantidades significativas de fibras e proteinas, alem de um potencial uso como nematicida. Uma vez que a producao mundial de oleo de mamona e de 1,5 milhao de toneladas-ano e a proporcao de producao de coprodutos com a producao de oleo e de aproximadamente 1:1, e fundamental encontrar um destino economicamente viavel para esses coprodutos. Alem dos destinos utilizados, uma rota com alto valor comercial seria o uso como racao animal, entretanto, isto nao ocorre devido a presenca da ricina. Esta, corresponde a uma proteina inativadora de ribossomos presente na semente da mamona, e e constituida por duas subunidades, a RTA e RTB, sendo a RTA a subunidade catalitica. Alem da problematica supracitada, a ricina e utilizada como arma biologica por terroristas e ativistas, logo, a inibicao do mecanismo de acao desta proteina e de grande interesse biotecnologico, sendo a RTA o alvo para sintese de inibidores. Neste aspecto, o metodo utilizado para busca de novos inibidores e o Atracamento Molecular (Molecular Docking). Este metodo avalia milhares de ligantes num curto intervalo de tempo, entretanto, apresenta baixa acuracia na predicao da afinidade de ligacao. Nesta perspectiva, a Dinamica Molecular Induzida (SMD), pode ser utilizada. Este metodo baseia-se na correlacao do perfil de forca medio necessario para desacoplar o ligante da proteina com a sua afinidade. A literatura cientifica tem relatado resultados promissores no uso dessa abordagem para discernir ligantes ativos de inativos. No presente estudo, alem de validacoes com inibidores de RTA conhecidos, foi avaliado o potencial de afinidade de 6 estruturas ineditas a formarem complexos com a RTA utilizando simulacoes SMD. Para isso, foi necessario recorrer a abordagens de Atracamento Molecular e Dinamica Molecular para obtencao e refinamento de novos complexos da RTA com os ligantes candidatos, para somente entao, serem submetidos a simulacao SMD (k = 2 kcal/mol/A², v = 0.005 nm/ps). Neste passo, alem do perfil de forca medio obtido a partir de multiplas simulacoes SMD independentes, foi mensurado a forca de ruptura e o trabalho medio realizado pela forca, este ultimo, apresenta uma relacao direta com a afinidade de ligacao pela igualdade de Jarzynski. Em suma, resultados de validacao da forca de ruptura e trabalho com os dados experimentais mostraram coeficiente de correlacao e determinacao com R = -0.992 e R² = 0.984 (forca de ruptura) e R -0.958 e R² = 0.918 (perfil medio do trabalho), respectivamente. Por meio destas validacoes, a avaliacao dos ligantes candidatos utilizando simulacoes SMD mostrou que 4 das 6 estruturas propostas, apresentam in silico, potencial de afinidade de ligacao para com a RTA.
  • JOSÉ MARREIRO DE SALES NETO
  • Efeito Anti-inflamatório da Piocianina em Macrófagos Murinos
  • Fecha: 29-nov-2016
  • Hora: 14:00
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  • A Pseudomonas aeruginosa e um bacilo ubiquo oportunista frequentemente associado a individuos portadores de patologias que causam imunodeficiencia, como a fibrose cistica. Essa bacteria produz um pigmento tipico, a piocianina, que representa o seu fator de virulencia majoritario, induzindo a geracao de especies reativas de oxigenio por celulas do hospedeiro, alem de induzir a inflamacao em varios tecidos e a apoptose em neutrofilos. No entanto, os protocolos utilizados para a observacao desses efeitos sao distintos entre si e nao se conhece o mecanismo de acao da piocianina em processos de inflamacao aguda. O objetivo desse trabalho foi compreender as interacoes entre a piocianina e os processos inflamatorios agudos, avaliando in vitro o efeito da piocianina na viabilidade celular e na producao de oxido nitrico, interleucina (IL)-1β e fator de necrose tumoral (TNF)-α por macrofagos peritoneais de camundongo ativados por lipopolissacarideo. Em adicao, foi avaliado in vivo o efeito da piocianina no modelo de peritonite induzida por zimosan em camundongo. A piocianina nas concentracoes de 50 ou 100 μM induziu a morte celular em 90 e 95% (p < 0,0001), respectivamente, enquanto a piocianina nas concentracoes de 1, 5 ou 10 μM nao foi capaz de afetar a viabilidade celular. O pigmento nas concentracoes de 5 ou 10 μM reduziu a producao do oxido nitrico em 26 % (p < 0.05) e 51 % (p < 0.0001), respectivamente, alem de diminuir os niveis da IL-1β (38 %, p < 0,001) e do TNF-α (48 %, p < 0,001) em macrofagos tratados com a piocianina na concentracao de 5 μM. No modelo in vivo, a piocianina na dose de 5 mg/kg nao interferiu na migracao dos leucocitos para o sitio da inflamacao. A reducao nos niveis do oxido nitrico, da IL-1β e do TNF-α induzida pela piocianina pode ser um mecanismo de fuga favoravel ao patogeno, capaz de reduzir a resposta inflamatoria do hospedeiro nas concentracoes avaliadas nesse trabalho, e esse efeito parece ser independente da interferencia na migracao celular.
  • JUCILENE PEREIRA DE SOUSA
  • PEIXE-ZEBRA (Danio rerio) TRANSGÊNICO PARA O GENE bmal1a: EFEITOS NO RELÓGIO MOLECULAR DO MÚSCULO ESQUELÉTICO
  • Fecha: 28-nov-2016
  • Hora: 14:00
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  • A maioria dos organismos apresentam ritmos circadianos em torno de um periodo de 24 horas que sao gerados por um mecanismo endogeno, o relogio molecular, que tem a capacidade de sincronizar-se com sinais ambientais. Este mecanismo tem fundamental importancia na homeostase dos tecidos que estao sob sua influencia. Dentre os genes que compoem a maquinaria do relogio molecular os genes clock e bmal sao os reguladores positivos do mecanismo desse relogio e apresentam expressao com perfil sigmoide em tecido como o musculo do peixe-zebra (Danio rerio), participando da ativacao de alguns fatores regulatorios miogenicos (MRFs – myoD, myog, myf5 e myf6), os quais possuem importancia para o desenvolvimento e diferenciacao do musculo. Apesar deste conhecimento, nao se sabe a importancia fisiologica do ritmo de expressao circadiana no musculo esqueletico de peixes. Neste sentido, o objetivo desse estudo foi investigar a taxa de transferencia do transgene para a prole; se a transgenia para o gene bmal1a no musculo esqueletico interferiu na sobrevivencia e crescimento dos peixes; e avaliar se a expressao dos genes bmal1a, clock1a e MRFs apresentaram diferencas na linhagem transgenica comparada a linhagem nao-transgenica (NT). Os fundadores (F0) foram obtidos apos a microinjecao do plasmideo e as larvas positivas foram observadas com coracao verde fluorescente. A F1 foi obtida a partir de cruzamentos entre peixes F0 e NT. Da mesma forma, F2 foi obtida a partir da F1, os quais foram utilizados no presente estudo. A analise da expressao genica das linhagens aos 11 meses de idade foi realizada utilizando a tecnica qPCR. A linhagem transgenica foi gerada com sucesso, transmitindo o transgene para a prole seguindo a heranca mendeliana. A sobrevivencia e crescimento da prole F2 nao apresentaram diferencas entre as linhagens, sendo 41,4±0% para a linhagem transgenica e 44,3±6% NT ate 30 dpf e 3.7±0.1 cm transgenicos e 3.8±0.2 cm para NT aos 11 meses de idade, respectivamente. Dentre os genes, o clock1a e o myog apresentaram diferencas estatisticamente significativas entre as linhagens com perfil circadiano em peixes NT, sugerindo que myog seja um gene controlado pelo relogio. Os demais genes apresentaram expressao constitutiva. De um modo geral, pode-se verificar que a expressao constitutiva do bmal1a nao apresentou alteracao na expressao do relogio molecular, desta forma, nao afetou a homeostasia do organismo, a sobrevivencia das larvas, bem como nao afetou o crescimento.
  • CAIO CÉZAR OLIVEIRA DE LUCENA
  • POTENCIAL ANTILEUCÊMICO DA LECTINA DE Cratylia floribunda E SUA CONJUGAÇÃO EM NANOTUBOS DE CARBONO
  • Fecha: 25-nov-2016
  • Hora: 14:00
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  • O cancer representa um dos maiores problemas de saude publica mundial, sendo a segunda causa de morte em todo o mundo. A busca por novos compostos que promovam a morte seletiva de celulas cancerigenas vem se tornando constante, como uma estrategia no tratamento da doenca. As lectinas constituem uma classe de glicoproteinas que reconhecem e se ligam especificamente a grupamentos de carboidratos expressos na membrana plasmatica. Os nanotubos de carbono vem ganhando bastante atencao por serem nanocarreadores promissores, possibilitando a conjugacao de diversas moleculas em sua superficie e aumentando a eficacia no transporte e a especificidade no reconhecimento celular. No presente trabalho, foi investigada a atividade citotoxica das lectinas Concanavalina A e CFL sobre linhagens de celulas de leucemia promielocitica cronica (K562) e monocitica aguda (THP-1). Para avaliar a viabilidade celular, foi utilizando o ensaio de reducao do sal de MTT, o qual avalia a capacidade metabolica da celula converter esse sal em cristais, foi observado um efeito citotoxico das lectinas nas duas linhagens testadas. Entretanto, so foi observada diminuicao da viabilidade apenas apos 72 horas de tratamento com as lectinas. As lectinas tambem foram citotoxicos as celulas endoteliais HUVEC, nao cancerigenas, porem essa resposta pode ter se dado pelo fato de essas celulas apresentarem diversos glicoconjugados na sua membrana. Foi tambem observado, por meio do teste de incorporacao do corante azul de tripam, o qual marca apenas em celulas com membrana rompida, que o conjugado lectina-nanotubo nao apresentou atividade diferente da demonstrada pela lectina sozinha frente as linhagens celulares, sugerindo que os nanotubos parecem nao melhorar o efeito das lectinas estudadas. Investigando qual a via de morte ativada pelas lectinas por meio de ensaios de citometia de fluxo, foi observado que as celulas marcaram positivamente para o iodeto de propideo, indicando que o tratamento de 72 horas causou danos a membrana celular. Entretanto, utilizando a sonda tetrametilrodamina metil-ester, foi visto que o tratamento com as lectinas causou a despolarizacao da membrana mitocondrial das celulas K562 e THP-1, fator relacionado a apoptose. Alem disso, a lectina CFL causou a parada do ciclo celular de celulas THP-1, promovendo um acumulo de celulas na fase G0/G1. Dessa forma, concluiu-se que as lectinas Concanavalina A e CFL demonstram efeito citotoxico as celulas leucemicas, possivelmente pela inducao da morte celular por apoptose nas celulas, devido a despolarizacao da membrana mitocondrial, possivelmente bloqueando a expressao de ciclinas e CDKs, promovendo parada no ciclo celular de celulas THP-1. Ja em celulas K562, o tratamento com a CFL por 72 horas pode estar ativando alguma via extrinseca de apoptose por receptor de membrana, levando a despolarizacao da mitocondria e consequentemente liberando fatores apoptogenicos, mas sem promover parada no ciclo celular. Desta forma, as lectinas estudadas demonstram um interessante potencial antileucemico. Palavras-chave: Cancer, lectinas, nanotubos, citotoxicidade, leucemia, apoptose
  • ANA GABRIELLA LUCENA DE PAIVA GUIMARÃES
  • PRODUÇÃO DE CONIDIOS E ENZIMAS HIDROLITICAS POR Beauveria bassiana (BALS) Vuillemin (DEUTEROMYCOTINA: HYPHOMYCETES) EM DIFERENTES SUBSTRATOS
  • Fecha: 25-nov-2016
  • Hora: 09:00
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  • Beauveria bassiana e um fungo entomopatogenico usado no controle biologico de insetos-praga na agropecuaria. Os custos de producao de seus conidios, em larga escala, no substrato padrao (arroz) utilizado atualmente onera o processo de producao. Encontrar alternativas como forma de minimizar os custos do processo, levou a necessidade de averiguar a eficiencia de substratos alternativos que, alem de propriedades nutricionais importantes, possuem grande disponibilidade e baixo custo. Os fungos entomopatogenicos produzem enzimas que estao envolvidas no processo de patogenicidade e virulencia. Porem, ao serem estimulados por substratos especificos podem produzir enzimas de interesse biotecnologico. Dessa forma, o objetivo deste trabalho foi analisar o potencial de diferentes substratos (fibra da algaroba, residuos de malte, sementes e fibras de acerola e bagaco da cana-de-acucar) para conidiogenese e producao enzimatica (fermentacao em estado solido) a partir de B. bassiana. Os experimentos para producao e viabilidade dos conidios foram realizados em triplicata, em erlernmeyer (250 mL) contendo 30 g de cada substrato, 0,3 µL da suspensao de conidios (1 x 106 conidios/mL), umidade de 70 ± 10% e temperatura (T = 29 ± 1° C). Apos 10 dias de incubacao, os substratos arroz (meio padrao) (2,00 x 106 conidios/g de substrato), malte A (1,22 x 106 conidios/g), malte B (1,75 x 106 conidios/g) e fibra de algaroba (2,36 x 106 conidios/g), pro¬porcionaram maior producao de conidios. A viabilidade dos conidios produzidos nesses mesmos substratos nao diferiu estatisticamente entre si, com porcentagem de germinacao de 99,96; 90,04; 93,17 e 98,21 %, respectivamente. A patogenicidade dos conidios de B. bassiana produzidos nos diferentes substratos foi avaliada no cupim do coqueiro. A taxa de mortalidade dos cupins infectados nao diferiu estatisticamente superando 80 % de mortalidade com excecao do grupo controle. A atividade enzimatica (celulolitica, amilolitica) foi determinada pelo metodo DNS (acido dinitrosalicilico). Os substratos utilizados na fermentacao em estado solido, malte A (1,178 ± 0,002 U/mL), malte B (2,392 ± 0,013 U/mL), fibra algaroba (0,596 ± 0,007 U/mL) e semente de acerola (0,964 ± 0,09 U/mL) apresentaram atividade amilolitica. A partir dessa analise, diferentes temperaturas foram avaliadas (30°, 40º, 50º, 60º, 65º e 70º C) para identificar a temperatura otima das enzimas amiloliticas produzidas nesse processo. Observou-se que as maiores atividades encontradas foram em temperaturas extremas, na faixa de 60º a 70ºC, sugerindo que essas enzimas sao termofilicas. O bagaco de cana de acucar apresentou maior atividade celulolitica (15,29 ± 0,07 U/mL) para CMcase e (2,58 ± 0,9 U/mL) para FPase devido a caracteristica de sua matriz celulosica, que difere dos demais substratos. A atividade proteolitica foi quantificada utilizando a azocaseina. A algaroba apresentou a maior atividade (0,514 ± 0,009 U/mL). Os substratos alternativos utilizados para crescimento e esporulacao de B. bassiana podem fornecer uma reducao de aproximadamente 50 % nos custos de producao de conidios de fungos entomopatogenicos utilizados no controle biologico de varios insetos-praga. Alem disso, apresentam-se como alternativa viavel para a producao de enzimas microbianas com aplicacao ampla em diversos processos biotecnologicos.
  • RICARDO BERNARDINO DE PAULA
  • EFEITOS DO 2-NITRATO-1,3-DI(OCTANOXI)PROPANO (NDOP) EM AORTA DE CAMUNDONGO C57BL/6
  • Fecha: 23-nov-2016
  • Hora: 09:00
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  • BERNARDINO-PAULA, R. EFEITOS DO 2-NITRATO-1,3-DI(OCTANOXI)PROPANO (NDOP) EM AORTA DE CAMUNDONGO C57BL/6. 2016. Dissertacao. (Centro de Biotecnologia, Universidade Federal da Paraiba, Joao Pessoa – PB, 2016) Nitratos organicos tem sido utilizados na terapia de desordens cardiovasculares pois sao doadores de oxido nitrico (NO) e podem mimetizar o papel do NO endogeno. O presente estudo foi desenvolvido com o objetivo de caracterizar o 2-nitrato-1,3-di(octanoxi)propano (NDOP), um nitrato organico sintetizado a partir da glicerina, como um doador de NO. A capacidade de liberacao de NO eliciada pelo NDOP foi avaliada em celulas de musculo liso vascular (CMLV) de arteria aorta e em ensaios de reatividade vascular em aneis de aorta toracica de camundongos (Mus musculus) C57BL/6 pre-contraidos com fenilefrina. Em adicao, foi tambem avaliada a tolerancia in vitro para o NDOP e a toxicidade aguda pre-clinica frente sua administracao via oral (v.o.) (300 e 2000 mg/kg) em camundongos femeas. O tratamento com NDOP aumentou os niveis de NO nas CMLV quando comparado as celulas estimuladas apenas com a sonda DAF (53,20 ± 1,61 vs.10,74 ± 0,86 a.u., n = 13 e 13, respectivamente, p < 0,05). O aumento de NO nas CMLV nao foi inibido pelo bloqueio da enzima oxido nitrico sintase endotelial (eNOS) utilizando L-NG-Nitroarginina (L-NNA, 100 μM) (52,80 ± 2,89 vs. 53,75 ± 2,18 a.u., n = 13 e 13, respectivamente). O NDOP (10-8 – 10-3 M) induziu efeito vasorrelaxante em aorta toracica de camundongo C57BL/6 com e sem endotelio funcional de forma semelhante (Emax = 102,6 ± 1,7% vs. 107,3 ± 7,5%, n = 8 e 6, respectivamente). A pre-incubacao com o bloqueador da NOS, L-NAME (100 µM) nao alterou a resposta vasorrelaxante em aneis com endotelio funcional (Emax= 112,5 ± 7,4%, n = 7). Por outro lado, o efeito vasorrelaxante do NDOP em aneis sem endotelio funcional foi diminuido pelo pre-tratamento com o sequestrador de NO, 2-(4-fenil)-4,4,5,5-tetrametilimidazolina-1-oxi-3-oxido (PTIO, 300 μM) (Emax= 75,7 ± 5,6%, n = 7, p < 0,05) e pelo PTIO juntamente com a hidroxicobalamina (Emax= 38,8 ± 4,6%, n = 4, p < 0,05). O uso do 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-ona (ODQ, 10 μM), um inibidor seletivo da enzima ciclase de guanilil soluvel (CGs) reduziu a resposta vasorrelaxante do NDOP (Emax = 22,2 ± 6.7%, n = 6, p < 0,05). Quando os aneis foram contraidos com a solucao de Krebs modificada com KCl 60 mM, a eficiencia maxima de relaxamento do NDOP foi atenuada (Emax = 80,4 ± 5,2%; n = 6, p < 0,05). O bloqueio inespecifico dos canais para potassio (K+) com solucao despolarizante de Krebs adicionada de KCl 20 mM reduziu a eficacia do relaxamento induzido pelo NDOP (Emax = 72,8 ± 3,4%; n = 6, p < 0,05). O uso de TEA (1 mM), 4-aminopiridina (1 mM) e Glibenclamida (10 µM) para bloqueio dos canais BKCa, KV e KATP, respectivamente, nao alterou o vasorrelaxamento induzido pelo NDOP. Entretanto, a pre-incubacao dos aneis com cloreto de bario (BaCl2, 30 µM), inibidor dos canais para K+ retificadores de entrada (KIR) reduziu a eficiencia de relaxamento induzido pelo NDOP (Emax= 73,7 ± 5,7%, n = 6, p < 0,05). A pre-exposicao dos aneis ao NDOP (10-4 M), durante 30 minutos, diminuiu o vasorrelaxamento induzido pelo composto em aneis com endotelio (Emax= 49,4 ± 3,5% vs. 102,6 ± 1,7%; n = 7 e 8, respectivamente, p < 0,05) e sem endotelio vascular (Emax= 44,1 ± 5,9% vs. Emax= 107,3 ± 7,5%, n = 6 e 6, respectivamente, p < 0,05). A administracao (v.o.) de 300 e 2000 mg/kg do NDOP em camundongos femeas demonstrou que o composto apresenta baixa toxicidade aguda pre-clinica e a DL50 foi aproximadamente 5000 mg/kg. Os resultados demonstram que NDOP atua como doador de NO em CMLV e que o vasorrelaxamento induzido pelo NDOP envolve a ativacao dos canais para K+ do tipo KIR e a participacao da CGs por meio da via NO-GMPc-PKG. Entretanto, apesar da baixa toxicidade aguda pre-clinica, o NDOP demostrou provocar tolerancia vascular in vitro. Palavras-Chave: nitratos organicos; oxido nitrico; canais para K+; vasorrelaxamento; toxicidade
  • ISABEL CRISTINA GUERRA GOMES
  • CARACTERIZAÇÃO MOLECULAR DOS SOROTIPOS DA DENGUE E CORRELAÇÃO DOS ÍNDICES PLUVIOMÉTRICOS E DOS CASOS DE DENGUE NA PARAÍBA, NO PERÍODO 2007-2015
  • Fecha: 01-nov-2016
  • Hora: 09:00
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  • A dengue pertence a familia Flaviviridae, genero Flavivirus e e uma infeccao viral transmitida por mosquitos. A doenca pode ser originada por qualquer um dos quatro sorotipos DENV que podem ser categorizados em: DENV-1, DENV-2, DENV-3, e DEN-4. O principal vetor e o Aedes aegypti, seguido pelo Aedes albopictus, sendo a propagacao da dengue atribuida a expansao de nichos dos vetores, bem como, pelo comercio global e aumento de viagens, a urbanizacao nao planejada, e ma gestao de residuos e agua. Dessa forma, o clima e considerado um fator importante na distribuicao temporal e espacial das doencas transmitidas por vetores. Apesar de ser a arbovirose mais prevalente em todo o mundo, a dengue tem sido negligenciada nos ultimos anos. Devido a recentes surtos epidemicos de diferentes arbovirus, como zika (ZIKV) e chikungunya (CHIKV) em todo o mundo, houve um alerta para as autoridades de saude em relacao a transmissao e propagacao das arboviroses. Assim, este estudo avaliou a classificacao e as caracteristicas clinicas dos casos de dengue, obtidas a partir das fichas de notificacao do Sistema de Informacao de Agravos de Notificacao (SINAN). A incidencia mensal de casos de dengue foi calculada pela populacao do ano/100.000 habitantes. A deteccao e genotipagem dos sorotipos DENV utilizou a tecnica nested RT-PCR, entre 2007-2015, e em 2016 para diagnostico diferencial foi realizada a tecnica PCR em tempo real (qPCR) para ZIKV e CHIKV. Alem disso, este trabalho analisou a correlacao entre a precipitacao pluviometrica e casos de dengue entre 2007 e 2015, na cidade de Joao Pessoa - PB, Nordeste do Brasil. Os dados de precipitacao foram coletados mensalmente atraves da sonda TRMM. A fim de identificar a influencia dos casos de chuva e dengue, foram considerados diferentes modelos de defasagem distribuida. Os resultados mostraram que, em relacao ao periodo de 2007 a 2015, foram notificados 55680 casos confirmados de dengue na Paraiba. Houve uma maior prevalencia de infeccao em individuos com idade entre 15-34 anos, em todos os anos estudados. Nossos resultados demonstraram a ocorrencia da correlacao (r ˃0.50) entre a precipitacao e casos de dengue, esses apresentaram um aumento nos primeiros meses apos o periodo chuvoso, demonstrando, pois resultados mais significativos ao considerar um lag de tres meses. O DENV-1 foi o virus mais prevalente (80,48%) em todos os anos estudados e a co-circulacao dos quatro sorotipos DENV foi observada em 2010, 2013 e 2014. Alem disso, observou-se a circulacao simultanea de casos de dengue, zika e chikungunya em 2016, demonstrando a predominancia dos casos de zika, seguido dos casos de chikungunya. Portanto, a caracterizacao molecular e a analise de correlacao entre numero de casos de dengue e precipitacao pluviometrica sao ferramentas importantes no futuro desenvolvimento de predicao nao so de casos de dengue, como tambem das arboviroses emergentes no estado da Paraiba, como a zika e a chikungunya. Palavras-chave: Epidemiologia, Dengue, DENV, Arbovirus, Fatores climaticos, Precipitacao pluviometrica.