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Item Caracterização de proteases secretadas por Sclerotinia sclerotiorum(Universidade Federal de Goiás, 2019-05-25) Alves, Elda Bueno; Silva, Silvana Petrofeza da; http://lattes.cnpq.br/6823998544968373; Silva, Silvana Petrofeza da; Andrade, Rosangela Vieira de; Faria, Fabrícia Paula deSclerotinia sclerotiorum is a fungus that infects many economically important plants. The production of oxalic acid, decreased the pH of the medium and the secretion of hydrolytic enzymes such as proteases, are determining factors for the infection of this pathogen. Proteases secreted by the fungus S. sclerotiorum degrade the cell wall proteins of the host and antifungal proteins secreted by the plant during the invasion. This study aimed to characterize molecular and biochemical proteases in S. sclerotiorum, during the invasion in bean plants (Phasheolus vulgaris L.) and in culture medium supplemented with pectin, the wall of bean extract and glucose as carbon sources. The secretion of these proteases was detected by enzymatic assays with specific inhibitors and buffered media at different pH. The level of expression of genes encoding for the protease aspartyl (aspS) and acid protease (acp1), respectively, were detected by qRT-PCR technique. Enzyme activity was detected in all sources of carbon, but higher levels were found when pectin was used as the sole carbon source. Tests with different proteolytic inhibitors showed that Pepstatin A specific inhibitor for aspartyl protease-inhibitor that has been the greatest action in culture media, indicating the probable secretion of aspartyl proteases. Of qRT-PCR experiments showed that the expression of genes aspS and acp1 was increased in S. sclerotiorum both in culture medium supplemented with pectin or extract of the cell wall of bean, as during the infection in bean seedlings. These results suggest that the fungus secret proteases during the infection of the host plant. And the different carbon sources evaluated, similar to components of the cell wall of host plants, are effective for the induction of gene expression (acp1 and aspS) in S. sclerotiorum. Thus, the proteases have been considered an important virulence factor in increasing the efficiency of fungal attack in plants.Item Efeitos metabólicos e reprodutivos da sobrenutrição pós-natal precoce em ratas Wistar provenientes da redução de ninhada durante a lactação: consequências na primeira e segunda geração(Universidade Federal de Goiás, 2019-04-17) Amaral, Keytiane de Jesus Viana; Cost, Renata Mazaro e; http://lattes.cnpq.br/5625298314637434; Gomes, Rodrigo Mello; http://lattes.cnpq.br/3121095341590269; Gomes, Rodrigo Mello; Costa, Renata Mazaro e; Corgosinho, Flávia Campos; Lima, Veridiana Mota MoreiraNutritional and environmental insults during perinatal period induces endocrine, functional and behavioral disorders in adulthood. Thus, it is necessary to assess the impact of infant feeding on the increased risk of obesity and other associated diseases, including in subsequent generations. The objective of the present study was to evaluate the metabolic and reproductive effects of early postnatal overnutrition in Wistar during lactation and the consequences in the second generation. Female Wistar rats (F0 generation) were distributed in two experimental groups: normal litter (NL, 9 pups, n=11) and reduced litter (SL, 3 pups, n=11) throughout the lactation period. On the 3rd postnatal day (PN3), litters SL were reduced to 3 pups/dam. After weaning (PN21), F1 females (NLF1 and SLF1) were fed with water and feed at will, and metabolic and reproductive parameters were analyzed in the adult (nulliparous), pregnancy and lactation phases. In the offspring of the F1 generation (F2 generation – GCF2 and GOF2), physical and sensory-motor, metabolic and reproductive parameters. In comparison to the NLF1 group, the SLF1 generation F1 group showed: a significant increase body weight in PN7, which was maintained during the adult phase, at the end of gestation and lactation; increased food intake during PN21 to PN70, without significant difference in the pregnancy and lactation phases; increase in the fat masses in the adult, pregnancy and lactation phases; increase in the masses of the liver in the lactation phase and of the pancreas in all phases; early of the onset of puberty and lower numbers of estrus; lower performance in sexual behavior; reduction in the number of corpus luteum, with no change in pre-implantation percentage, post-implantation losses and number of live male and female fetuses; fewer live births, both male and female; greater maternal care; reduction of serum estradiol concentration; increase in serum triglyceride concentrations and reduction of HDL cholesterol in pregnancy and lactation, with no change in total cholesterol levels; increase in adult glucose. In the histopathological analyzes there was also an increase in the area of adipocytes and the thickness of the adipose layer of the mammary glands; reduction in the number of growing follicles and increase in the number of cystic and atretic follicles in ovaries; endometrial glands with inflammatory cells and degenerating in the uterus. The GOF2 offspring, compared to the GCF2 offspring, presented higher body weight from birth to PN21, as well as higher milk intake in PN8. In the evaluation of the physical and sensory-motor development, the GOF2 offspring presented advance of the sensory-motor parameters (righting reflex, negative geotaxis, cliff avoidance) in males and females. However, the auditory startle reflex was later in males and females, with no significant difference in smell and free-fall righting parameters. Regarding the physical parameters, the eruption of the incisors was anticipated and the opening of the eyes delayed, without significant difference in the unfolding of the ears. The onset of puberty was anticipated in males and females. In conclusion, early postnatal overnutrition, acting as a factor of metabolic programming, promoted obesity, metabolic and reproductive changes in F1 generation rats and, consequently, also promoted metabolic and reproductive changes and in the physical and sensory-motor development of F2 offspring.Item Envolvimento do núcleo pré-óptico mediano (MnPO) na recuperação cardiovascular induzida pela infusão de salina hipertônica em animais submetidos ao choque hemorrágico(Universidade Federal de Goiás, 2014-04-16) Amaral, Nathalia Oda; Custodio, Carlos Henrique Xavier; Pedrino, Gustavo Rodrigues; http://lattes.cnpq.br/1155446449250341; Oliveira, André Henrique Freiria de; Ferreira Neto, Marcos Luiz; Ferreira, Patrícia Maria; Castro, Carlos Henrique deIn recent decades, several studies have demonstrated that hyperosmolarity induced by hypertonic saline infusion (HS) it’s a benefit for hypovolemic hemorrhage treatment. The median preoptic nucleus (MnPO) is known to receive information from central osmoreceptors and peripheral afferents about plasma osmolarity changes, reflexively modulating autonomic and neuroendocrine adjustments, primarily through its projections to the paraventricular nucleus (PVN). The present study aim to determine MnPO involvement in cardiovascular recovery induced by HSI in rats subjected to hemorrhagic shock (HC). Wistar rats (250 - 300 g) were prepared to record mean arterial pressure (MAP), heart rate (HR), renal blood flow (RBF) and aortic (ABF). The renal vascular conductance (RVC) and aortic (AVC) were calculated through the ratio between RBF and ABF with MAP, respectively. Hemorrhagic shock was induced by blood withdrawal over 10 min until the MAP reached approximate values of 60 mmHg. The sodium overload by infusion HS (3 M NaCl, 1.8 ml ∙ kg-1 body mass index) was made 2 min after the nanoinjection (100 nL) of GABA agonist muscimol (4 mM experimental group 1 - EXP 1 ), α-adrenergic antagonist phentolamine (13 mM ; experimental group 2 - EXP 2) or isotonic saline (NaCl, 0.15 M, control group - CON) in MnPO. This resulted in HC CON (n=6) MAP reduction (98.4 ± 5.3 to 62.2 ± 1.1 mmHg after 20 min HC, p<0.05), a decrease in RVC (- 59.4 ± 9.2%, 20 min after HC, p<0.05) and did not alter the AVC (-11.5 ± 10.5%, 20 min after HC) and HR (387.2 ± 12 to 351.7 ± 13 bpm after 20 min HC). HC promoted in EXP 1 (n=6) MAP reduction (98 ± 5.4 to 61 ± 0.7 mmHg after 20 min HC, p<0.05), a decrease in RVC (-64.8 ± 10.9%, 20 min after CH, p<0.05) and CVA (-32.3 ± 4.4%, 20 min after HC, p<0.05) and did not alter HR (389 ± 23.9 ± 17.1 to 360 bpm after 20 min HC). In EXP 2 (n=6) HC resulted in a MAP reduction (102.0 ± 4.2 to 62.0 ± 1.1 mmHg, 20 min after HC, p<0.05), a decrease in CVR (-27.6 ± 5.8% after 20 min HC, p <0.05), CVA (-4.5 ± 4.1% after 20 min HC, p<0.05) and HR (387 ± 14 to 347 ± 7.4 bpm after 20 min HC). HS infusion enabled MAP restoration (105.2 ± 3 mmHg, 60 min after infusion of HS, p<0.05), did not alter HR (400 ± 18.4 bpm, 60 min after infusion of HS) raised the RVC to baseline xi levels (-14.6 ± 14.2%, 60 min after infusion of HS, p<0.05) and reduced AVC (- 27.4 ± 4.3%, 60 min after infusion HS, p<0.05) in CON. HS infusion in EXP 1 was not able to restore MAP (54 ± 3.8 mmHg, 60 min after infusion of HS, p<0.05) and RVC (- 48.1 ± 9.7%, 60 min after infusion of HS, p<0.05), did not alter HR (361 ± 15.3 bpm, 60 min after infusion of HS) and was able to promote an increase in AVC similar to baseline (-23.2 ± 10.6%, 60 min after infusion HS, p<0.05) levels. In EXP 2, HS infusion enabled MAP restoration (89 ± 3.3 mmHg, 60 min after infusion of HS, p<0.05) but this return to baseline was delayed and occurred only 50 min after HS infusion (88 ± 3.3 mmHg), HR return (379 ± 6.5 bpm, 60 min after infusion of HS) and RVC to basal levels (-16.1 ± 8.9%, 60 min after infusion HS, p<0.05) and an increase in AVC 10 min after HS infusion (20.3 ± 6.4%, p<0.05), which was restored to levels similar to baseline at registration end (-15.7 ± 6.2%, 60 min after infusion of HS, p<0.05). Together, the results obtained in this study showed that MnPO plays an important role in cardiovascular recovery induced by HS infusion in HC cases. Furthermore, the cardiovascular adjustments involved in this resuscitation seem to depend partly on adrenergic neurotransmission in this nucleus.Item Estudo da toxicidade e genotoxicidade induzidas por diferentes nanopartículas in vivo(Universidade Federal de Goiás, 2012-02-27) Andrade, Laise Rodrigues de; Cunha, Kênya Silva; http://lattes.cnpq.br/6124135410387685; Cunha, Kênya Silva; Lee, Chen Chen; Irazusta, Silvia PierreThe rapid advancement of nanotechnology has created a vast array of nanoparticles promising for industrial, energy and environmental sectors. Furthermore, many biological applications have been proposed for the nanoparticles, such as transport of genes and drugs for disease treatments, including cancer and infections. So, it is important to clarify if nanoparticles may represent a hazard to the environment and human health. In this study, we investigated the toxic potential and ability to induce DNA damage of different nanoparticles: four species of carbon nanotubes (NTC, NT, CS e MW), silver (AgNP) and nanoceria (CeO2-NP) nanoparticles. It was employed the somatic mutation and recombination test (SMART) in Drosophila melanogaster that detects the loss of heterozygosity of two genetic markers involved in the metabolic pathways of wing hairs formation - multiple wing hairs (mwh) and flare3 (flr3). In larvae, the proliferating of wings imaginal disc cells can produce hairs with mutant phenotypes expressed on the adult wings. Using the standard cross, third-stage larvae were treated with different concentrations of the nanoparticles until pupal stage. The wings of adult flies were examined microscopically for the identification of phenotypic abnormalities. In all concentrations the survival rates were higher than 90%, indicating the absence of chronic toxicity for nanoparticles evaluated. Using the conditional binomial test, the results of different treatments were compared with the respective negative control (distilled water), demonstrating no significantly increase in the NTC, NT and CS mutation and recombination frequencies (p>0.05) in mwh/flr3 genotype. In all carbon nanotubes tested, only the two higher concentrations of nanotubes MW (0.4 and 1 mg/mL) were able to induce genetic changes, mainly by mitotic recombination. The concentration of 10 mg/mL AgNP also promoted changes in the DNA and 61% of the phenotypic abnormalities were caused by recombination. The nanoceria was able to produce genotoxic effects at all concentrations tested (0.64-10 mg/mL). Overall, the mutational events were predominant, ranging from 46 to 72% of the total genotoxic effect induced by nanoceria, showing no dose-response relationship. In conclusion, our results demonstrated that carbon, silver and cerium dioxide nanoparticles have different genotoxic potential in D. melanogaster, so, another studies should be performed before any clinical and/or industrial application of nanoparticles.Item Avaliação da exposição aguda e sub-letal ao glifosato (N-fosfometil-glicina) e ao AMPA (ácido amino-metil-fosfônico) em brânquias e fígado de Poecilia reticulata com o emprego de biomarcadores moleculares e morfológicos(Universidade Federal de Goiás, 2013-02-15) Antunes, Adriana Maria; Bailão, Alexandre Melo; http://lattes.cnpq.br/5415221996976886; Morais, Simone Maria Teixeira de Sabóia; http://lattes.cnpq.br/6723881044959716; Morais, Simone Maria Teixeira de Sabóia; http://lattes.cnpq.br/6723881044959716The present research aimed to investigate the acute toxicity of glyphosate (Nphosphomethyl-glycine), the active ingredient of Roundup®, as well as of aminomethylphosphonic acid (AMPA), its main degradation product, in the teleost fish Poecilia reticulata, commonly known as guppies. Thus, were performed tests Lethal Concentration mean (LC50) and behavioral, histopathology and proteomics analysis. Male and female P. reticulata presented median lethal concentration (LC50) at 96h the values of 68.78 and 70.87 mg/L for glyphosate and 180.00 and 164.32 mg/L for AMPA, respectively, characterizing them as practically non toxic compounds to guppies. However, the results of behavioral analyses showed that both products generate stressing situations to the fish, causing behavioral changes such shock against the tank walls, hypoactivity, irregular swimming, swimming on the surface of water, darkening of the body, aggressiveness, and lethargy. Histopathological and morphometric analyses of liver and gill of guppies exposed to sublethal concentrations of glyphosate (35mg/L) and AMPA (82 mg/L) indicated that both organs were affected . The gills of fish exposed to the tested products presented higher reaction rates to progressive changes, such as proliferation of the interlamellar epithelium and partial and total fusion of secondary lamellae. Liver presented higher reaction rates to regressive changes, such as steatosis, pyknotic nuclei, and necrosis. The proteomic analysis showed that Glyphosate induces differential liver proteins in P. reticulata. The image analysis allowed measuring 225 spots in the control group and 180 spots in the group exposed to Glyphosate. Of this total, 88 spots were exclusive to the control group and 43 spots were exclusive of the treated group. Of the 137 spots matched between control and treated groups 13 were down regulated and 8 were up regulated due to sublethal exposure to Glyphosate. The behavioral, histopathological and proteomic changes observed indicated that, although considered to have low toxicity to animals in the guides that evaluate toxicity of chemicals, both glyphosate and AMPA promote that are deleterious to animal health, indicating the need to establish new parameters or modify concepts indicating the need to use several biomarkers, beyond assessing the mortality rate, to establish a more reliable criteria for toxicity parameters.Item Avaliação da atividade anti-inflamatória docarotenoide bixina em ratos submetidos ao modelo de colite ulcerativa induzida por ácido trinitrobenzenossulfônico(Universidade Federal de Goiás, 2017-09-28) Araújo, Joniel Mendes de; Ferreira, Anderson Luiz; http://lattes.cnpq.br/9495326031912899; Ferreira, Anderson Luiz; Leal, Geraldo Sadoyama; Moreira, Karla GraziellaCrohn's disease (CD) and ulcerative colitis (RCU) are the major and most prevalent inflammatory bowel diseases (IBD). The lack of effective treatment makes the search for new drugs to treat IBD extremely important. Bixin, a carotenoid present in the species Bixa orellana L., appears as a possible candidate for presenting several biological activities among them antibacterial, antioxidant and anti-inflammatory. The objective of this study was to evaluate the intestinal anti-inflammatory effect of Bixin in experimental TNBS-induced colitis in rats. For the experimental protocol Wistar rats (n = 10) were divided into 7 groups: Saline group (without colitis), TNBS group (with colitis) and groups treated with Bixina gavage at doses of 1, 3, 6, 9 and 12mg / kg. Colitis was induced according to the procedure described by Morris et al. (1989). Euthanasia was performed by CO2 overdose followed by removal of the colon for macroscopic and histological analysis. The results demonstrated that the presence of adhesions and diarrhea was decreased in the groups treated with Bixina. The macroscopic analysis of the colon evidenced the presence of severe damage to the colonic mucosa, with edema, deep ulcerations and hemorrhage in the TNBS group. The Bixina-treated group (12 mg/kg) significantly attenuated the areas of ulcerative lesion and macroscopic score when compared to the TNBS group. Animals receiving Bixin (12 mg/kg) also had a decreased mass / length ratio of the colon compared to the TNBS group. The histological characteristics of the colon of the TNBS group showed severe signs of inflammation, necrosis and edema. However, the Bixin group (12 mg/kg) showed better cell organization and less intestinal epithelial damage when compared to the TNBS group. The results suggest a protective effect of the carotenoid Bixin against experimental colitis induced by TNBS. However, new studies must be performed to verify the mechanisms involved in this activity.Item Variabilidade genética e avaliação de sensibilidade a fungicidas em sclerotinia sclerotiorum proveniente de cultivo irrigado de feijoeiro(Universidade Federal de Goiás, 2015-03-27) Arboleda, William Andrés López; Silva, Silva Petrofeza da; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4793542D5Sclerotinia sclerotiorum is a phytopathogenic fungus that infects more than 400 plant species, including common bean. Genetic variability studies in connection with phenotypic traits of agronomic interest are important to drive the control strategies against this pathogen. The aims of this study were: to evaluate the genetic variability, fungicide sensitivity, aggressiveness and to determine the proportion of MAT (Mating Type) alleles of 79 isolates of S. sclerotiorum distributed in four populations from common bean. Two populations represented a single sampling location in two different times (2000 and 2013 growing seasons). To evaluate the fungicide sensitivity a cell viability test based on the alamarBlue dye using mycelial growth was standardized. Dose-response curves for fluazinam, procymidone and benomyl were estimated using this test and were compared with dose-response curves estimated by the mycelial growth inhibition on PDA plate and the FRAC (Fungicide Resistance Action Committee) protocol. Despite the differences to assess the fungicide sensitivity between the three methods, the dose-response curves showed similar trends for the three fungicides. The fungicide sensitivity assessment at the four populations showed low sensitivity to benomyl in the Planaltina population. Furthermore, this population presented a principally clonal population structure, with a haplotype represented by 18 out of 20 isolates. Significant population differentiation in all pairwise comparisons of phi, except the comparison between EV_2013-NH, was detected. Five genetically homogeneous groups were inferred by the DAPC analysis. No group was conformed by isolates from the four populations. Only two haplotypes between the two populations from the same sampling location were shared. The hypothesis of random mating was rejected at the four populations; however this hypothesis was not rejected at the two major populations inferred by the DAPC analysis. The screening of mating type locus showed a dominance of Inv+ isolates and a high proportion of Inv+/Inv- isolates (presumable heterokaryons).Item Ocorrência de mutações em loci ligados ao cromossomo Y na prole nascida de indivíduos expostos à radiação ionizante(Universidade Federal de Goiás, 2009-05-13) ARRUDA, Jalsi Tacon; CRUZ, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985In September 1987, in Goiânia-GO, Brazil, one of the most serious radiological accidents occurred at a radiotherapy unit involving a source of cesium-137. An area of 2,000 m2 was contaminated and 249 people were exposed, both externally and internally, to substantial doses of ionizing radiation, resulting in four fatalities due to acute radiation syndrome. The current study examined the occurrence of possible mutations in the Y chromosome of the exposed men and their male offspring divided into two groups: A) eight accidentally exposed men and eight boys; B) twelve occupationally exposed men and sixteen boys; and the control group with 8 men and 8 boys not exposed. DNA was isolated from peripheral blood lymphocytes and 30 loci (SRY, AMELY, ZFY, AZFa-Prox1, SY83, AZFa-Prox2, SY86, SY85, SY84, USP9Y, SY87, DBY, AZFa-Dist1, 12f2, AZFa-Dist2, UTYpe, SY106, SY124, SY127, SY134, SY135, SY143, SY1197, SY1291, SY1125, SY1054, YDAZ3, SY254, SY255, RH65618) were amplified by the polymerase chain reaction. All DNA tests had a probability of paternity of at least 99.99%. All analyzed individuals amplified STS; however, 4 fathers (8.4%) and 8 sons (21.2%) in group A, and 3 fathers (7.1%) and 3 sons (63.3%) in group B showed mutations. The total mutation rates were 0.11. The first generation of the accidentally exposed group showed 7 mutations in SY86, 12 mutations in SY84, and 1 mutation both in 12f2 and SY135. The first generation of the occupationally exposed group showed 2 mutations in SRY, AMELY, AZFa-Prox1, AZFa-Prox2, SY86, SY85, SY84, USP9Y, SY87, AZFa-Dist1, UTYpe, SY106, SY124, SY127, SY134, SY135, SY143, SY1125, SY1054, YDAZ3, SY254, SY255, and RH65618; and 4 mutations in 12f2. In the control group, only one son showed an SY84 deletion. Recombination events between repetitive regions are possibly the cause of the high incidence of de novo mutations in the Y chromosome. The mutations were possibly generated by intrinsic mechanisms that could have been increased by the ionizing radiation from cesium-137. The exposure to ionizing radiation from cesium-137 can be detected in offspring of exposed individuals, and the mutation rate can be attributed to radioactive exposure.Item Análises in vivo e in vitro de interações intermoleculares da Beta-1,3- glicanosiltransferase 1 de Paracoccidioides brasiliensis(Universidade Federal de Goiás, 2010-01-28) BAILÃO, Elisa Flávia Luiz Cardoso; SOARES, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637The cell wall of pathogenic microbes acts as an initial barrier that is in contact with hostile environments. Besides functioning as a mechanical barrier, it harbours an immunogenic macromolecules arsenal. One of the ways that proteins can be associated to the cell wall, it is through GPI anchor. The hydrophobic C-terminal end of the β-1,3-glucanosyltransferase enzyme of the human pathogenic fungus Paracoccidioides brasiliensis is characteristic of GPI anchored proteins. The β-1,3-glucan assembling and rearrangement are essential since this molecule acts as a scaffold to support cell wall proteins and polysaccharides. In the thermodimorphic fungus P. brasiliensis, β-1,3-glucan is found predominantly in mycelium form and α-1,3-glucan is predominant in the yeast form. In this work, it was screened possible protein-protein interactions performed by β-1,3-glucanosyltransferase 1 of P. brasiliensis (PbGel1p). To obtain these results, a P. brasiliensis cDNA library was screened with PbGel1p using the Saccharomyces cerevisiae two hybrid system. In addition, pull-down assay was used as an in vitro complementary technique to isolate proteins that interact direct or indirectly with PbGel1p. It was screened 38 gene products using two hybrid system and it was identified 3 proteins using the pull-down assay associated with mass spectrometry. The PbGel1p role in the cell wall maintenance and remodeling was indicated through the analysis of screened interactions, like alpha-glucosides permease, acid phosphatase, GDSL lipase, septin, actin, tubulin, HSP90 and pyruvate kinase. Furthermore, nuclear localization of PbGel1p and its role in the locus-specific transcriptional silencing were suggested based on such interactions: Qde2 argonaute, transcription elongation factor spt6, others transcription factors and ATP-citrate synthase. Therefore, this study indicated, for the first time, that PbGel1p has multiple location and it participates either in roles classically described for glucanosyltransferases, as the cell wall remodeling, or in recently described functions for this family of proteins, as the locus-specific transcriptional silencing.Item Diversidade, relações filogenéticas e perfil de expressão gênica de transportadores MFS em Trichoderma spp(Universidade Federal de Goiás, 2021-04-21) Barbosa Filho, Jomal Rodrigues; Georg, Raphaela de Castro; http://lattes.cnpq.br/3971276154067590; Georg, Raphaela de Castro; Coelho, Alexandre Siqueira Guedes; Steindorff, Andrei SteccaThe Trichoderma genus comprises filamentous fungi with mycoparasitism ability and high tolerance to abiotic stresses, such as the presence of metals. In both conditions, cellular transporters are vital to nutrients absorption and metallic ion extrusion. Among those, the Major Facilitator Superfamily (MFS) correspond to the largest superfamily of secondary transporters found in every organism, however a deep understanding of those is needed in Trichoderma spp. The present work aimed to identify the MFS transporters from 32 species of this genus, as well as to study their phylogenetic relationship and gene expression profiles. Global MFS identification were performed in silico using the Trichoderma spp. reference proteomes available on the Mycocosm database from Joint Genome Institute (JGI), by searching for annotated aminoacid sequences using the BlastP tool. The sequences were categorized in their respective families, according to reference sequences from the Transporter Classification Database (TCDB), and maximum likelihood phylogenetic trees were estimated for each of the families, using the IQTREE software. Lastly, MFS genes expression profiles were investigated during mycoparasitism and presence of metals, using RNA-Seq public data. 8,467 putative MFS were identified in Trichoderma spp., with a mean of 264 ± 51 sequences, and this number was higher than those found in other organisms. Mycoparasitic species presented a higher number of MFS. 55.5% of the sequences showed 12 transmembrane helices and 85.5% showed molecular weights between 50 and 70 kDa, with a mean of 58 kDa. In total, 24 MFS families were identified in the analyzed species, and the families regarding to nutrition, maintenance of homeostasis and cell defense showed a higher number of members. We did not observed enrichment of families in any species analyzed. In the phylogenetic inference the sequences grouped according to their functions and not to their lineage, moreover, SHS and GPH families showed subdivisions. In T. harzianum, besides a higher number of differently expressed genes during mycoparasitism, a higher number of upregulated genes was also observed in this condition compared to those in the presence of metals. In the presence of cadmium, we observed a higher downregulation of MFS genes than in the presence of aluminum. We also observed that ACS and DHA2 families were mostly differentially expressed during mycoparasitism 24 h, whilst FHS and PHS were mostly differentially expressed during the presence of aluminum 1.5 and 3.0 mg/mL, and MCT during the presence of cadmium 2.0 mg/mL. This work shed light on the description of MFS transporters and their functions during some of the biotic and abiotic stresses faced by Trichoderma spp. in the course of its development on environment.Item Avaliação de critérios de compatibilidade entre pares de primers para otimização de sistemas multiplex de genotipagem(Universidade Federal de Goiás, 2010-01-12) BARBOSA, Ana Clara de Oliveira Ferraz; COELHO, Alexandre Siqueira Guedes; http://lattes.cnpq.br/0840926305216925The progress of Molecular Biology and Genetics provided the appearance of several molecular markers that detect the genetic polymorphism directly at DNA. Among these markers are the microsatellites (SSR), which are distinguished by their high degree of polymorphism. The use of these markers for individual genotyping has evolved into multiplex systems, which allow many SSR fragments to be detected and analyzed simultaneously. Currently there are several articles in literature discussing the criteria to be used in the primer design for use in PCR, as well as various softwares are available for this end. However, there are few studies and tools for the analysis of compatibility between pairs of primers for use in multiplex systems, where multiple fragments are simultaneously amplified using PCR. This paper evaluated different criteria for compatibility between pairs of primers. A set of 74 combinations of pairs of primers, involving the amplification of 94 SSR loci were evaluated in duplex systems. The same combinations were evaluated according to different criteria, including the degree of complementarity between primers, the magnitude of differences of denaturation temperatures (Tm) and the tendency to annealing between pairs of primers based on the Gibbs free energy resulting from the association between them. The comparison between the different criteria allowed the identification of a set of criteria with positive predictive value equal to 94%. These criteria were implemented for use in a software called Multiplexer, which from the analysis in sequence of pairs of primers, suggests compatible combinations for use in multiplex genotyping systems. Using this tool can significantly reduce the costs related to laboratory activities for genotyping using PCR.Item Avaliação da atividade da invertase de Saccharomyces cerevisiae imobilizada em polianilina sobre o caldo de cana(Universidade Federal de Goiás, 2010-02-19) BARBOSA, Eduardo Fernandes; FERNANDES, Kátia Flávia; http://lattes.cnpq.br/9737543228759171This work describes the immobilization of invertase on chemically synthesized polyaniline and activated with glutaraldehyde (PANIG) for production of invert syrup from sugarcane juice. Free invertase activity present in crude extract (E.B.) obtained from cells of Saccharomyces cerevisiae, was characterized for an evaluation of interferents present in the extract on enzyme activity (optimum conditions: temperature 50 ° C, pH of 4.5 in sodium acetate buffer 0.1 mol L-1 and reaction time of 10 minutes, with an activity of 11.31 ± 0.36 EU mL-1). We tested some parameters optimization of enzyme immobilization, such as amount of enzyme, immobilization time, pH and temperature of immobilization. The optimal immobilization was obtained in buffer sodium acetate 0.1 mol L-1 pH 4.5, immobilization time of 1 hour at 50°C and 169.55 EU mg-1 PANIG. The efficiency of immobilization was 0.86. The stability of the system PANIG-Invertase was tested against the storage time and thermostability, and after 75 days storage in buffer sodium acetate 0.1 mol L-1 pH 4.5 was obtained for 94% of initial activity with only 17% retained for the free enzyme. The immobilized invertase didn t change the optimal conditions compared to the free, but the immobilized was more stable in adverse conditions such as pH below and above optimum conditions showed an increase in thermostability. Some features of the hydrolysis product were evaluated (water activity, viscosity and color), compared to the sugarcane juice in nature, showing that the reactors allowed changes in sugarcane juice that expand the possibilities for using syrup obtained in the production of sweets, ice cream and syrups rich in fructose. The high stability of the system tested, along with its high retention of activity strongly suggests the use of the system in reactors.Item Morfo-anatomia e fitoquímica de Cymbopogon densiflorus (Steud.) Stapf e Cymbopogon nardus (L.) Rendle (Poaceae: Panicoideae)(Universidade Federal de Goiás, 2007-09-28) BARBOSA, Lília Cristina de Souza; PAULA, José Realino de; http://lattes.cnpq.br/3191837532986128; REZENDE, Maria Helena; http://lattes.cnpq.br/5093753722360659The genus Cymbopogon Sprengel belong to the Poaceae family and it has 40 species distributed in Tropical and Subtropical Africa, Asia and Australia, although some species went introduced in America. Many species of this genus are cultivated for the extraction of essential oil, from their leaves, with large medicinal, food and industrial importance. The species in focus, Cymbopogon densiflorus (Steud.) Stapf and C. nardus (L.) Rendle are originated from Africa and Asia, respectively. This research had as objective, to broaden the knowledge about the species C. densiflorus and C. nardus, by the morphological and anatomy studies from leaves and culms, phytochemical analysis and essential oil analysis from the leaves. Anatomical studies have been of relevant importance to the pharmacognosy researches, mainly for the identification of many vegetal raw materials. Several times, these raw materials are known by the same popular name or then, they are commercialized with contaminated agents or with other parts of the specie. Through of anatomical analysis, it was checked commons characters, such as leaf lamina and sheath amphistomata, stomatas with guard cells dumbbell and subsidiary cells dome-shape, rares in adaxial surface and abundant in abaxial surface, predominated in intercostal zones. The adaxial and abaxial surfaces had long cells and short cells: cork and dumb-bell and cross-shaped silica cells, these last it is placed in costal zones; macro-hairs and micro-hairs abundant in abaxial surface. In the leaf lamina, bulliforms cells are presents in adaxial surface, they were alternated with fibers in the costal zones and the mesophyll is homogeneous with chlorenchyma radiated to the bundle sheaths and arm cells with walls invaginated that they determined the intervenal distance by one or three cells, characterized Kranz anatomy. Bundle sheaths collateral, of 1st, 2nd and 3rd orders with single vascular bundle sheaths. The cap region is constituted by sclerenchyma and the epidermis has silica cells. However, both species had different anatomical features, as the form of midrib, in the leaf laminas; C. densiflorus showed colourless parenchyma cells in the mesophyll of leaf sheaths, that they do not exist in C. nardus. In the culms, numbers of metaxylem vessels in the each side of protoxylem vessels in vascular bundles: 1, in C. densiflorus, 2 and 3, in C. nardus; and the presence of sclerenchyma cylinder and fistula in C. nardus, absent characters in C. densiflorus. Moreover, in C. densiflorus, while C. nardus showed these characters. The preliminary phytochemistry analysis C. densiflorus and C. nardus leaves evidencied flavonoids, saponins, coumarins and traces of cardioactive glycosides. In the essential oil analysis, C. densiflorus leaves showed trans-p-mentha-1(7),8-dien-2-ol, trans-p-mentha-2-8-dien-1-ol, cis-carveol and cis-p-mentha-2-8-dien-1-ol as majority constituents; while C. nardus leaves had geraniol, citronellol and citronellal. The anatomical characters observed can be important to the taxonomic determinations of species studied, in the genus. Through the results found, it verifies the phytotherapics potential of both species. Future researches in isolation and purify of the secondary metabolites, pharmacologics and toxicologics analysis of extracts and of the essential oil, it will be important to assure the therapeutic efficiency of these.Item Análise dos níveis de poligalacturonases e glucanases expressas durante os processos de interação patogênica e saprofítica de Sclerotinia sclerotiorum(Universidade Federal de Goiás, 2008-05-30) BARBOSA, Silvio Romero Costa; SILVA, Silvana Petrofeza da; http://lattes.cnpq.br/6823998544968373The fungus Sclerotinia sclerotiorum can interact with a great range of vegetable species as well as to obey to the discharge especificity and patogenic specialization . It is capable to digest the cellular wall of host plants using for such a series of biochemical mechanisms and morfogenetics that optimize the invasion. Several enzymes are produced during the interaction plant-host and among them they stand out the family of the poligalacturonases (PGs) and them beta glucanases. PGs catalyze the hydrolysis of the connection glycosídic bond - 1,4 and them beta glucanases liberate glucans and oligossacarídeos during the hydrolysis. Our work tried to characterize, besides the pH, the action of these enzymes, as well as the gene expression during the interaction with bean (Phaseolus vulgaris) and under different cultivation means, pectin 1%, wall extract 1% and glucose 1%. The activities were measured by the method DNS where the amount was measured of you sugar reducers in the middle and the gene expression through electrophoretic profiles analyzed after the technique of RT-PCR. The results showed variation of the activity of PGs in the interaction being the middle with pectin with larger expression of them. Them beta 1,3 and beta 1,4 glucanases were expressed in both culture means proposed, however there was larger production of beta 1,3 during the invasion. The variation of the expression of such enzymes in different culture means suggests complexity of specific biochemical roles for your production raising new approaches for the recognition of the roads that they promote the development of the diseaseItem Expressão heteróloga do gene Hxyn2 do fungo Humicola grisea var. thermoidea em Pichia pastoris: Produção e purificação da enzima HXYN2r e aplicação em testes de panificação(Universidade Federal de Goiás, 2008-05-30) BASTOS, Fernando Medeiros; FARIA, Fabrícia Paula de; http://lattes.cnpq.br/3739169267521003Endoxylanases are the main group of enzymes involved in the hydrolysis of xylan. This enzymes have application in industrial proposes, like as drink, food, feed, clothes industries and for bleaching cellulose paper pulps. In bread making the xylanases have been used to improve processing and product quality of loaf, leading soft dough and loaf with larger volume as well as an improved crumb structure. The xylanolitic enzymes produced by filamentous fungi constitute an enzymatic pool with distinct activities which make their use in industrial process more difficult, the obtainment of recombinant microorganisms constitutes a strategy to achieve suitable enzymes to industrial process. The thermophilic fungus Humicola grisea var. thermoidea has proved to be a good source of xylanolitic enzymes. The gene Hxyn2 from H. grisea codes a xylanase with 23 kDa that belongs to G/11 family of glycosil-hydrolases. Its cDNA was cloned in the vector pHILD2 and expressed in yeast Pichia pastoris under the control of the promoter AOX1. The transformants were chosen trough genetic stability and capacity to produce and secrete the enzyme HXYN2r into culture medium. The purified HXYN2r showed a high xylanolitic activity with an optimum temperature of 60 ºC and an optimum pH value of 6.5. The aim of this project was the production, purification and application of HXYN2r enzyme in bread making tests. The production in the optimized medium obtained 100 mg of active xylanase per liter of medium BMMY-U. This quantify of enzyme presented 40% of the total proteins in culture supernatants. The proteins of culture supernatants was concentrated by liofylization and fractionated by into a chromatographic column of gel filtration Sephacryl S-100. The purified xylanase presented specific activity of 2250 U/mg and the purification profit was 6.4%. The 23 kDa protein was confirmed by the activity on Zymogram assay. The pure xylanase was added at the rate of 45 and 90 U/Kg of wheat flour in the bread making tests. In this rates it was not observed any effect of xylanase in specific volume either in crumb structure. The HXYN2r enzyme was partially purified by a heat treatment (45 min at 60 °C) and was concentrated by liofylization and a yield of 17.9% was obtained. The partially purified HXYN2r was added at the rates of 500, 1500, 3000, 4500 and 6000 U/kg of wheat flour. The added xylanase improved the specific volume and the crumb structure, but any effect was observed in the moisture content of the loaf. The best result was the rise of 16.0 % in loaf specific volume with the dose of 3000 U/kg of wheat flour. This effect was similar to the obtained with a commercial xylanase added to the dough in equal dose. The results presented suggest that the xylanase from H. grisea can be used in bread making to improve specific volume.Item Planejamento, síntese e avaliação farmacológica de novos candidatos a protótipos de fármacos anti-inflamatórios(Universidade Federal de Goiás, 2017-03-03) Batista, Daniel da Costa; Menegatt, Ricardo; http://lattes.cnpq.br/8354030864254626; Menegatt, Ricardo; Neves, Josilane Sabbadini; Oliveira, Guilherme Roberto deInflammation is defined to be a beneficial response, dynamic and adaptive of body against any aggressor agent, be it of chemical, physical or biological nature, aimed at neutralizing this agent, as well as tissue repair. During the inflammatory process are produced and/or released biomolecules that will promote the maintenance of this process as well as the emergence of events called "cardinal signs of inflammation" as pain, heat, swelling and redness which if not properly addressed can result in loss of injured tissue function. Histamine, a biogenic amine synthesized from L-histidine amino acid exerts its physiological action by binding to histamine receptors called H1R, H2R, H3R and H4R which are characterized by belonging to the class of GCPR. In recent years there has been reported a number of research on the role of the H4R receptor in inflammatory processes, since it is expressed in immune cells membrane regulating the chemotaxis induced by histamine and the synthesis of compounds such as the prototype JNJ7777120 (8), capable of modulating such receptor once there are no drugs that acts front this mechanism of action available in the market. In search a line of research aimed at planning, synthesis and pharmacological evaluation of new candidates prototype anti-inflammatory drugs, we describe in this paper the design, synthesis and pharmacological evaluation of new N-arilheterocycles derivatives (34a-34p and 34'a), designed from the rings bioisosterism strategy from prototype JNJ7777120 (8), with possible future therapeutic applications. In this sense the LQFM182 derivative (34a) was subjected to investigation of their anti-inflammatory properties in the mice paw edema models and pleurisy induced by carrageenan, as well as evaluation of MPO activity and quantification of IL-1β and TNF-α cytokines was able of promoting a reduction of 87% in the levels of the latter, coming to be most effective in dose used when compared to dexamethasone, which promoted an 82% reduction in TNF-α levels.Item Tratamento com aceturato de diminazeno ou angiotensina-(1-7) em ratas hipertensas gestantes atenua disfunções cardiovasculares na prole(Universidade Federal de Goiás, 2019-04-22) Bessa, Amanda de Sá Martins de; Castro, Carlos Henrique de; http://lattes.cnpq.br/6354834854727314; Castro, Carlos Henrique de; Costa, Renata Mazaro; Fonseca, Silvia Carolina GuatimosimIt has been proven that harmful stimuli during gestation can promote deleterious outcomes in offspring. Furthermore, previous studies have observed that the Renin-angiotensin system (RAS) can play a role in this pathological process. TheACE2/Ang-(1-7)/Mas axis presents several protective actions in the cardiovascular system. Thus, the aim of this study was to investigate whether the treatment with diminazene aceturate (DIZE), a putative ACE2 activator, or with Angiotensin-(1-7) during the pregnancy could attenuate the development of cardiovascular dysfunctions in the adult offspring of spontaneously hypertensive rats (SHR). For this, pregnant SHR received DIZE or Ang-(1-7) during the gestation. The SBP was measured in the male offspring by tail-cuff plethysmography. Thereafter, the left ventricular contractile function and coronary reactivity were evaluated by the Langendorff technique. Aortic vascular reactivity was also evaluated by isolated vessel in an organ bath. Samples of the LV were collected for histology and Western blot assay. Maternal treatment with DIZE (SHR: 181 ± 2.03 vs. SHR DIZE: 166.6 ± 0.35 mmHg, P <0.05) or Ang-(1-7) (SHR: 167.3 ± 1.79 vs. SHR Ang-(1-7): 153.8 ± 1.75 mmHg, P <0.05) during pregnancy attenuated the increase of the SBP in adult offspring. Additionally, the treatments reduced the cardiomyocyte diameter (SHR: 16.3 ± 0.11 vs. SHR DIZE: 12.9 ± 0.95 μm, P <0.05) (SHR: 16.6 ± 0.12 vs. SHR Ang-(1-7): 14 ± 0.10 μm, P <0.05) and degradation of the extracellular matrix (SHR: 15.6 ± 0.96 vs. SHR DIZE: 9.0 ± 0.61%, P <0.05) (SHR: 17.2 ± 1.27 vs. SHR Ang- (1-7): 10.3 ± 0,73%, P <0.05) in LV. The maternal treatment with DIZE and Ang-(1-7) improved the coronary vasodilation induced by bradykinin in isolated hearts. The expressions of AT1, Mas, ACE, ACE2, ERK Total, P-ERK, TNF-α, Collagen I, SOD, and Catalase in LV were not modified with Ang-(1-7), but this treatment decreased the AT2 (SHR: 0.90 ± 0.03 vs. SHR Ang- (1-7): 0.74 ± 0.05, P <0.05) expression. These data show that the treatment with DIZE or Ang-(1-7) during gestation promoted beneficial effects in attenuating hypertension and cardiac remodeling in adult offspring.Item Avaliação da atividade genotóxica e antigenotóxica de Celtis iguanaea (Jacq.) Sargent em bactérias e camundongos(Universidade Federal de Goiás, 2010-02-24) Borges, Flávio Fernandes Veloso; Lee, Chen Chen; http://lattes.cnpq.br/4621907105842007Celtis iguanaea (Jacq.) Sargent is a member of the Cannabaceae family and popularly known as esporão de galo. Ethnobotanical surveys of cerrado native plants show that the tea leaves of C. iguanaea is used in traditional medicine for body aches, asthma, cramps, poor digestion, urinary infections, kidney disfunctions, as a stimulant or as a diuretic.The purpose of the present work was to evaluate the possible cytotoxic, genotoxic and antigenotoxic effects of the leaves aqueous extract of C. iguanaea by the prophage λ induction test (SOS Inductest) and the micronucleous test in mice bone marrow. The SOS Inductest was performed according to Moreau (1976), using lysogenic WP2s(λ) and indicator RJF013 strains derived from Escherichia coli. WP2s(λ) cultures were treated with different doses of the extract (0.5, 1, 2.5, and 5 mg) (evaluation of genotoxicity) or co-treated with a single dose of MMC (0,5 mg) and different doses of extract (assessment of antigenotoxicity). Then, the treated culture was added to the indicator strain (RJF013) and both were poured on plates in LB(1/2) medium. For the evaluation of cytotoxicity, cultures of WP2s(λ) treated with different doses of extract were diluted in M9 buffer and plated on LB. The micronucleus test was carried out according to Schmid (1975). To assess the genotoxic activity, animals were treated with Celtis iguanaea extract (100, 300 and 500 mg/kg concentrations). To evaluate the antigenotoxic activity, the same doses of extract were treated simultaneously with 4mg/Kg of MMC. The frequencies of micronucleated polychromatic erythrocytes (MNPCE) were evaluated at 24h and 48h exposure period except the negative control (24h). The cytotoxicity was assessed by the polychromatic and normochromatic erythrocytes ratio (PCE/NCE). In both tests the obtained results demonstrated anticytotoxic activity and absence of genotoxicity and cytotoxicity of the Celtis iguanaea leaves extract. The extract also showed partial antigenotoxic activity in bacteria and antigenotoxic activity in mice.Item Avaliação dos aspectos fisiológicos, bioquímicos e moleculares no feijoeiro comum ( phaseolus vulgaris) com o uso do indutor biótico trichoderma harzianum contra sclerotinia sclerotiorum(Universidade Federal de Goiás, 2012-09-10) Brandão, Renata Silva; Lobo Junior, Murillo; Ulhoa, Cirano José; http://lattes.cnpq.br/8368469162867277The fungus Trichoderma spp., is a microorganism naturally found in soil, which presents an important ecological function, as part of the decomposition and mineralization of plant residues, contributing to the availability of nutrients to plants. It is considered also a natural biofungicide, which reduces up to 100% the chances of achieving any fungus culture. The fungus Trichoderma is a fast growing, hence the advantage of use as a biocontrol agent in large scale, produces a variety of antifungal metabolites, including antibiotics and enzymes that degrade the cell wall of certain pathogens, is related to growth promotion in plants and the induction of resistance. The fungus Sclerotinia sclerotiorum is the etiological agent of the disease known as white mold in different cultures, such as soft rot, white mold, tipping pre-emergent and post-emergent plants. It is estimated that about 300 plant species are susceptible to S. sclerotiorum, such as the common bean (Phaseolus vulgaris) and soybean (Glycine max), both of great economic importance. Thus, this work needs to evaluate physiological, biochemical and molecular in common bean using the inductor Trichoderma harzianum against Sclerotinia sclerotiorum. Three isolates of T. harzianum and one isolate S. sclerotiorum were selected for inoculation in bean plants. In plants inoculated with isolates of T. harzianum 475/02 was possible to observe higher values related to the physiological aspects, as the plants inoculated with T. harzianum 479/01 had the highest enzyme activities in the R5 stage. Treatments with more pathogen T. harzianum showed the highest expression of the genes of interest, demonstrating greater efficiency when the two fungi act together.Item Prospecção e caracterização de peptaibols produzidos porlinhagem de Trichodermaasperellum(Universidade Federal de Goiás, 2014-08-08) Brito, João Paulo Cabral de; Ulhoa, Cirano Jose; http://lattes.cnpq.br/8368469162867277; Ulhoa, Cirano Jose; Monteiro, Valdirene Neves; Fernandes, Kátia FláviaTrichoderma genus corresponds to free-living filamentous fungi, including species able to act as biological control agents against pathogenic fungi. It is believed that this ability is due to the synergy of several mechanisms, including a wide variety of secondary metabolites produced by these organisms. Among them, there is the production of peptaibols, an antibiotic peptide group characterized by the presence of non-proteinogenic amino acids such as α-aminoisobutyrate (Aib), presence of N-terminal modifications and amino alcohols in the C-terminal region. These peptaibols have amphipathic nature, allowing the formation of ion channels in lipid membranes, which are believed to be related to its antimicrobial activity. This study aimed to draw a profile of production and identify of secreted peptaibols from Trichoderma asperellum TR356 strain. The fungi was grown in TLE with 0.3% glucose medium for 5 days, 120 rpm in the dark. Liquid medium filtrate was used as metabolites source. These extracts were subjected to high performance liquid chromatography (HPLC) and subsequent analysis by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF). The results indicate the production of two classes of peptaibols for this T. asperellum strain. Primary structure of two Asperelines (A and E) and five Trichotoxins (T5D2, T5E, T5F, and T5G 1717A) has been elucidated. Most of these peptaibols had been previously described in T. viride and T. asperellum marine strains. This is the first report of some of these compounds being produced by a T. asperellumstrain from soil. Future analysis will be necessary to elucidate the three-dimensional structures and their activity against pathogens.