Programa de Pós-graduação em Ciências Biológicas

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    Ação antimicrobiana de enzimas hidrolíticas produzidas por Trichoderma asperellum e imobilizadas em blendas de polímeros biodegradáveis.
    (Universidade Federal de Goiás, 2011-01-28) SILVA, Barbara Dumas Santos; ULHOA, Cirano Jose; http://lattes.cnpq.br/8368469162867277; FERNANDES, Kátia Flávia; http://lattes.cnpq.br/9737543228759171
    The hydrolytic activity of enzymes produced by Trichoderma asperellum, immobilized biodegradable films, as growth inhibitor of microorganisms was tested. The inhibitory activity was demonstrated on Aspergillus niger, Penicillium sp. and Sclerotinia sclerotiorum, microorganisms usually related to the attack and/or food contamination at the field or packaged. We used two polymer blends with different compositions, cassava starch and poly-butylene adipate-co-terephthalate (Ecoflex®, BASF Chemical Company) and other composed for polyvinyl alcohol (PVA) and polysaccharide cashew gum (PEJU). T. asperellum was induced to produce enzymes involved in the attack mycoparasite (N-acetylglucosaminidases, β-1,3-glucanases, chitinases and proteases) by the addition of crude chitin in the growth medium. The enzymes produced in major quantity were N-acetylglucosaminidase and chitinase. The pool of enzymes produced in the experiments was then used for immobilization tests. The immobilization process was performed in films by two methods: covalent and ionic bonding. In both methods, the presence of immobilized hydrolytic enzymes resulted in reduced growth of microorganisms, but the covalent immobilization of the results were more expressive. S.sclerotiorum was the microorganism most sensitive, followed by A. niger and Penicillium sp. To confirm the action of hydrolytic enzymes produced by T. asperellum and evaluate the effects they produce cell wall of microorganisms and other structures, the films with enzyme immobilized by covalent bonding were subjected to scanning electron microscopy. The structures most affected were hyphae and spores. Overall, the synergistic action of all enzymes produced by T. asperellum, reduced the growth of microorganisms when immobilized on the surface of the films Starch-Ecoflex® and PVA-PEJU. Moreover, the polymer blends tested exhibited desirable characteristics for future use in food packaging and most importantly, also provide efficient systems for the immobilization of enzymes.
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    Ação biológica in vitro de tiossemicarbazonas derivadas de canfeno e limoneno em células de melanoma humano (SK-MEL-37)
    (Universidade Federal de Goiás, 2013-05-16) Passos, Débora Cristina Silva dos; Oliveira, Cecília Maria Alves de; Guillo, Lídia Andreu; http://lattes.cnpq.br/3401436781775091
    Melanoma is a type of cancer that arises from melanocytes and is notoriously resistant to radiation and chemotherapy. The thiosemicarbazones are synthetic compounds with marked biological properties such as antibacterial, antiviral, antiprotozoal and antitumor and previous studies have demonstrated cytotoxic activity against the human melanoma cells, so in this study, we evaluated the antiproliferative activity, the enzymatic activity of Caspases 2, 3, 6, 8, 9, the effect on the cell cycle gene expression levels of caspases 2, 3, 6, 8, 9, Apaf-1 and microscopic morphological changes in human melanoma cells (SK -MEL-37) twenty one monoterpene derived from natural thiosemicarbazone (-) - camphene: camphene, benzaldehyde, benzophenone, menthone, ethyl pyruvate, p-nitroacetophenone, pchloroacetophenone, p-methoxyacetophenone, p-methylacetophenone, p fluoracetofenona-phidroxiacetofena, furan, 3-methoxy-4-hydroxybenzaldehyde, p-fluorbenzaldehyde, 2- hydroxybenzaldehyde, cinnamic aldehyde, thiophene-2-carboxaldehyde, 1-H-imidazole-4- carboxaldehyde, tiossemicaroazida and six montoterpeno natural R-(+)-limonene: benzaldehyde, thiosemicarbazide, o-nitro, m-nitro, p-nitro, p-hydroxy and p-dimethylamino. The values found for the inhibitory concentration for 50% of cells (IC50) were between 12 μM and 55 μM. The percentage of cells in phase and in phase G0/G1 decreased SG2 / M increased after forty-eight hours of incubation with benzaldehyde thio-camphene, limonene thio-benzaldehyde, m-nitro, p-hydroxy and thiosemicarbazide increased indicating that the growth inhibitory effect might be also due to arrest of cells at S-G2/M phase. We observed increased activity of caspase 3 (m-nitro thio-limonene), 6 (camphene thio-benzaldehyde and p-hydroxy thio-limonene) and 8 (thio-benzaldehyde limonene). Late apoptotic features were detected in 62% of cells treated with benzaldehyde thio-camphene and morphological changes typical of apoptosis were visualized by fluorescence microscopy and scanning electron microscopy (SEM) after treatment with benzaldehyde thio-camphene chosen due to their low IC50 value (12 mM). It was observed gene expression of caspases 2, 3, 6, 8 and Apaf-1 in cells treated with benzaldehyde thio-camphene indicating the participation of these enzymes in the anti-proliferative effect observed. Our results indicate that the thiosemicarbazones derivatives can inhibit proliferation, regulate cell cycle, induce apoptosis of human melanoma cells (SK-MEL-37) and could be an candidate for future preclinical in vivo studies.
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    Ação da fosfolipase B extracelular de Paracoccidioides brasiliensis na interação ex vivo com macrófagos alveolares
    (Universidade Federal de Goiás, 2010-03-26) SOARES, Deyze Alencar; SILVA, Silvana Petrofeza da; http://lattes.cnpq.br/6823998544968373
    Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of the most prevalent systemic mycosis in Latin America, paracoccidioidomycosis. The phospholipase B (PLB) enzyme is considered an important virulence factor in this dimorphic fungus, involved in the immune response of the host-pathogen interaction. Our objective was to determine whether a P. brasiliensis (Pb18) PLB is involved in adhesion / internalization of yeast and evasion of host immune responses. The effect of PLB was analysed using specific inhibition of PLB (alexidine dihydrochloride) and pulmonary surfactant in an ex vivo model (Pb18) of alveolar macrophage (MHS cells) infection. PLB enzyme assays and real time RT-PCR (qRTPCR) analysis of genes differentially expressed in the process of evasion: plb1 (phospholipase B1), icl1 (isocitrate lyase) and sod3 (Cu, Zn dismutase) and immune responses: clec2 (C-type lectin domain 2), cd14 (cluster of differentiation 14), tlr2 (toll-like receptor 2), nfkb (nuclear factor kappa B), nkrf (NF-kappaB repressing factor), il1β (inteleukin-1β) and tnfα (tumor necrosis factor alpha) were carried out using selective inhibition of PLB activity and pulmonary surfactant. The levels of cytokines inteleukin 10 (IL-10), IL-12 and TNF-α) were also determined by ELISA. PLB activity under adhesion conditions of P. brasiliensis (Pb18) to alveolar macrophage cells was found at high levels up to 6 hours post-infection. In the conditions of exposure to pulmonary surfactant and alexidine dihydrochloride, PLB activity and the level of transcripts of genes related to phagocytosis and inflammatory response were measured. We found that PLB activity had an influence on the phagocytic activity of alveolar macrophages. Alexidine dihydrochloride (0,25 μM) selectively inhibited PLB activity by 66% and decreased significantly the adhesion and internalization of yeast on MHS cells. Genes involved in phagocytosis (trl2 and cd14) and inflammatory response (nrkf, tnfα and il1β) were down-regulated in the presence of the PLB inhibitor. In contrast, the PLB activity and internalization of fungal yeast cells increased significantly in the presence of pulmonary surfactant (100 μg/mL) and genes such as clec2, important for effective phagocytosis by MHS cells, and the pro-inflammatory inhibitor (nkrf) were up-regulated. Also, the pulmonary surfactant did not alter cytokine production, while alexidine dihydrochloride decreased the levels of IL-10 and increased the levels of IL-12 and TNF-α. In addition, through simultaneous analyses of gene expression for the pathogen, P. brasiliensis, we found upregulation of the genes sod3, icl1 and plb1, required for the evasion of alveolar macrophages. P. brasiliensis PLB is important for the binding and internalization of yeast at macrophage surfaces. The specific effect of inhibiting PLB enzyme activity indicates that adhesion may be facilitated indirectly via fatty acid release from phospholipids of the membrane of host cells. This is the first study to show that PLB activity may modulate immune responses to P. brasiliensis infection.
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    Alteração no padrão de expressão de genes associados ao perfil leucemogênico da leucemia linfóide aguda infantil: antes e depois da quimioterapia de indução
    (Universidade Federal de Goiás, 2013-01-13) Minasi, Lysa Bernardes; Silva, Daniela de Melo e; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985
    The ALL is a malignant disorder that originates from a precursor cell lympho-hematopoietic system compromised for the development of B or T lymphocyte lineage.The precursor cell acquisition by a series of genetic abnormalities alters the normal process of maturation leading to cellular differentiation arrest and leukemic clone proliferative advantage on cells of hematopoietic tissue. More than 50 recurrent genetic alterations have been identified in the ALL. These often involve genes known or putative role in the development of lymphocytes and in the case of leukemogenesis.In this study, the variation in the expression of molecular markers was analyzed using PCR methodology array on 16 children with ALL before treatment and at the end of induction chemotherapy (D +28). These patients were diagnosed in HAJ and SCMG, from May 2012 to January 2013. Samples of bone marrow or peripheral blood were sent to NPReplicon-PUC-GO. In the present study we observed a negative correlation between gender and immunophenotype (p = 0.016), females have a greater association with immunophenotype B and less associated with immunophenotype T. We also observed a positive correlation between immunophenotype and age (p = 0.04), immunophenotype and marker CD10 + (p = 0.03), immunophenotype and risk group (p = 0.015) and marker CD10 + and risk group (p = 0.043). Before treatment the gene RUNX1 met with increased expression by 5.0 times compared to the control group and after induction chemotherapy was observed a reduction in its expression. The expression pattern of the gene TAL1 showed significant decrease, with the exception of post-treatment analysis showed that an increase in its expression. A positive correlation was observed between the expression of BAX and BCL2 (r = 0.94 and p <0.0001. We demonstrated a significant difference in gene expression pattern of ALL at diagnosis and after induction therapy. We conclude our observation regarding the gene expression profile in patients with ALL enrolled in this study, but to define a panel of molecular markers is necessary to evaluate several other genes involved in the process of leukemogenesis in ALL with the help of other methodologies.
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    Alterações cardiovasculares induzidas pela administração aguda e crônica do protótipo de fármaco LQFM-21
    (Universidade Federal de Goiás, 2015-02-26) Trindade, Neidiane Rosa; Menegatti, Ricardo; Pedrino, Gustavo Rodrigues; http://lattes.cnpq.br/1155446449250341
    This study aimed to investigate the effects hypotensive (LQFM-21), trying to identify a pharmaceutical potential for this new drug prototype. Male Wistar spontaneous hypertensive rats (SHR) and wistar rats were used to evaluate the effect of LQFM-21 on mean arterial pressure (MAP), heart rate (HR), renal vascular conductance (RVC), arterial vascular conductance (CVA) and baroreflex sensitivity index (BI). The administration of LQFM-021 (0.05, 0.1, 0.2, 0.4 mg / kg, iv.) Decreased MAP (-9.0 ± 1.4, -21.5 ± 2.9, ± 2.8 - 26.7 mmHg respectively); increased RVC (12.0 ± 1.9, 23.2 ± 3.2, 26.8 ± 6.3, 34.8 ± 5.9%, respectively) and CVA (17.3 ± 1.7; 27 8 ± 2.6, 32.7 ± 4.8 and 42.2 ± 3.4%, respectively) in anesthetized normotensive rats. In SHR, also found reduction of MAP (-16.9 ± 2.8; -19.4 ± 2.5; -22.0 ± 3.8 -17.2 ± 6.4 mmHg, respectively), HR (-11.2 ± 0.8 bpm at a dose of 0.04 mg / kg) and an increase in RVC (22.2% at a dose of 0.2 mg / kg) and CVA (12.2 (12.2 ± 2.1, 13.8 ± 5.6, 18.2 ± 2.3, 15.6 ± 3.1% at doses of 0.05, 0.1, 0.2, 0.4 mg / kg, respectively). Both the acute, oral as the oral administration chronicle of LQFM-21 at a dose of 15 mg / kg reduced the MAP in animals wistar 89.9 ± 1.9 mmHg and 136.4 ± 4.9 mmHg SHR without changing BI in wistar animals with the infusion of phenylephrine -2.3 ± 0.32 vs 1.9 ± 0.1 mmHg / bpm and SHR -1.9 ± 0.2 vs. -1.9 ± 0.3 mmHg / bpm, with infusion nitroprusside in wistar 3.2 ± 0.31 vs 3.0 ± 0.04 bpm / mmHg and 3.9 ± 0.6 vs SHR 3.0 ± 0.04 bpm / mmHg. Together our results demonstrated hypotensive effects of LQFM- 21. However further studies are needed to identify the mechanism of action and toxicity of this drug prototype.
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    Alterações cardiovasculares induzidas pelo aumento da ingestão de sal ou de sacarose
    (Universidade Federal de Goiás, 2017-01-17) Moreira, Marina Conceição dos Santos; Pedrino, Gustavo Rodrigues; http://lattes.cnpq.br/1155446449250341; Pedrino , Gustavo Rodrigues; Santos, Fernanda Cristina Alcântara dos; Rebelo, Ana Cristina Silva; Reis, Ângela Adamski da Silva; Mendes, Elizabeth Pereira
    Cardiovascular diseases (CVD) and their complications are the main causes of death around the world nowadays. Among these diseases, the hypertension stands out because of its great prevalence and multifactorial characteristic. Several studies demonstrate that, among other factors, the increased intake of industrialized food (rich in both salt and sugar) contribute to the pathophysiology of salt-dependent hypertension and obesity-related hypertension. However, the high sucrose intake induced cardiovascular modifications still need to be clarified. Based on these information, we sought to evaluate the effects of high salt intake of different durations during the post-natal period and of high sucrose intake in adulthood on cardiovascular parameters in adult animals. The present study sought to determine the cardiovascular and autonomic effects of salt and sucrose overload in adult animals. We evaluated the mean arterial pressure (MAP), the heart rate (HR) and the baroreflex sensitivity of the animals submitted to salt overload after weaning and to sucrose overload in adulthood and the cardiac morphology and Rostral Ventrolateral Medulla (RVLM) sensitivity after salt overload. 21 days old male Wistar rats received hypertonic saline solution (NaCl 0,3M) for 30 or 60 days (experimental groups). The control groups were maintained with tap water for equivalent period. After the treatment, the groups were maintained with tap water and food for 15 days (recovery period). A distinct group of adult animals was submitted to sucrose overload for 30 days. At the end of the experimental protocols, the animals were chronically cannulated. 24h after the surgery, the basal cardiovascular parameters and its modifications induced by baroreflex/chemoreflex stimulation were recorded. The baroreflex index was calculated as the ratio between HR and MAP changes after each infusion of phenylephyne and sodium nitroprusside. There were no differences between baseline MAP and HR of the animals treated during 30 days compared to the age-matched control animals (cont.: 118.2 ± 3.7 mmHg vs. exp.: 112.3 ± 4.0 mmHg; cont.: 404.0 ± 10.6 bpm vs. exp.: 374.7 ± 9.1 bpm). However, these animals presented diminished baroreflex sensitivity compared to the control group (BI: cont.; -2.441 ± 0.359 bpm/mmHg vs. exp.: -1.434 ± 0.086 bpm/mmHg, p<0.05). The animals submitted to high salt intake for 60 days after weaning presented increased MAP (cont. 98.6 ± 2.6 mmHg vs. exp. 117.7 ± 4.2 mmHg; p<0.05) and HR (cont. 365.4 ± 12.2 bpm vs. exp. 392.5 ± 10.3 bpm; p<0.05) compared to their control group. Moreover, the baroreflex sensitivity was diminished in those animals (cont. -1.83 ± 0.04 bpm/mmHg vs. exp. -1.24 ± 0.19 bpm/mmHg; p<0.05). The type I collagen relative frequency increased in the hearts of adult animals after 60 days after salt overload (cont.: 15.74 ± 0.61% vs. exp.: 19.79 ± 1.26%; p<0.05) but no differences were observed in the cardiomyocyte cross-sectional area. Furthermore, the pressure response to glutamate nanoinjections into the RVLM was increased in the animals treated for 60 days compared to the control group (cont.: Δ15.47 ± 2.56 mmHg vs. exp.: Δ34.31 ± 4.65 mmHg; p<0.05). The sucrose-treated animals presented increased MAP and HR compared to the control group (cont.: 102.5 ± 1.4 mmHg vs. exp.: 111.3 ± 0.9 mmHg; cont.: 334.7 ± 7.3 bpm vs. exp.: 371.6 ± 4.7 bpm; p<0.05). Furthermore, they presented diminished hypotension-induced baroreflex sensitivity (cont.: 5.0 ± 0.1 bpm/mmHg vs. exp.: 4.0 ± 0.1 bpm/mmHg; p<0.05) and increased pressure response after chemoreflex stimulation (cont.: 14.9 ± 1.9 mmHg vs. exp.: 29.2 ± 5.5 mmHg; p<0.05). Our results allow us to conclude that changes in the reflex regulation of blood pressure induced by salt overload after weaning precede the cardiovascular effects induced by such protocol, indicating that baroreflex sensitivity modifications can occur independently on permanent blood pressure changes. Moreover, salt overload after weaning promotes increases in RVLM sensitivity in adult animals. Furthermore, sucrose overload promotes MAP, HR and chemoreflex sensitivity elevation, whereas it promotes diminish of baroreflex sensitivity in adult rats. Taken together, our results allow us to conclude that changes in salt or sucrose intake produce permanent modifications in the arterial pressure control and such modifications are, in part, induced by changes in neuronal regulation of blood circulation.
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    Alterações cardiovasculares induzidas por supernutrição durante a lactação em ratos Wistar na idade adulta
    (Universidade Federal de Goiás, 2019-02-18) Ferreira Junior, Marcos Divino; Castro, Carlos Henrique de; http://lattes.cnpq.br/6354834854727314; Gomes, Rodrigo Mello; http://lattes.cnpq.br/3121095341590269; Gomes, Rodrigo Mello; Mendes, Elizabeth Pereira; Matafome, Paulo Nuno Centeio
    Metabolic syndrome, closely related to obesity and its comorbidities, has become a worldwide pandemic, with increasing projections each year. The role of adipose tissue in the metabolic syndrome is not limited to the accumulation of lipids but also to the secretion of adipokines, which can control food consumption and energy expenditure. In obese individuals, there is a change in adipokines profile, from anti-inflammatory to pro-inflammatory. Inflammatory status in adipose tissue leads to insulin resistance, increased adipocyte differentiation and macrophages recruitment. In addition, the angiotensinogen (AGT) produced by adipose tissue may correspond to more than 30% of plasma levels of this substance, which increases in case of adipocyte hypertrophy. AGT is a key part of the Renin Angiotensin System (RAS), and is directly linked to circulating Angiotensin II (ANG II) levels, and the latter is linked to obesity-induced cardiovascular changes. Several studies have proposed to evaluate the effects of obesity on cardiovascular parameters, but the majority in animals with diet-induced obesity. Here, we aimed to evaluate the effects of postnatal overnutrition-induced obesity (PNO), due to litter reduction (SL), in metabolic and cardiovascular parameters in adulthood. In our study, SL group developed hyperphagia, greater adiposity and body weight, and dyslipidemia. Also, important cardiovascular alterations were developed in SL animals, such as hypertension, left ventricular hypertrophy, endothelial dysfunction, and increase of oxidative stress related proteins. However, through in vivo and ex vivo cardiac function analyzes, our results suggest that the hearts of animals in the SL group present neurohumoral compensation of their function, which leads to overload and may progress to heart failure. Taken together, our data may help in the elucidation of mechanisms by which obesity-induced cardiovascular changes occur, increasing the possibility of discovering new targets for treatment.
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    Alterações dos parâmetros de comportamento de ratos tratados com peptídeo rico em prolina da serpente Bothrops jararaca, Bj-PRO-7a
    (Universidade Federal de Goiás, 2018-05-16) Turones, Larissa Córdova; Ianzer, Danielle Alves; http://lattes.cnpq.br/7609262674053858; Custódio, Carlos Henrique Xavier; http://lattes.cnpq.br/0207928273284808; Pansani, Aline; Gomes, Rodrigo Mello
    The Bj-PRO-7a, a heptapeptide isolated and identified from Bothrops jararaca (Bj) venom, evoke potent therapeutic effects, as antihypertensive effect, natriuretic and diuretic effects, promotion of angiogenisis and vasodilatation. The effects of heptapeptide are independent of Angiotensin Converting Inhibitor (ACE), possibly dependent of Muscarinic Receptors subtype 1 (M1R) activation and oxido nitric pathways. Also, the Bj-PRO-7a actions upon central nervous system still need to be evaluated. Thus, the aims of this study were: i) to assess the effects of acute administration of Bj-PRO-7a upon behavior; ii) to reveal mechanisms involved in the effects of Bj- PRO-7a upon locomotion/exploration, anxiety and depression-like behaviors. For this purpose, adult male Wistar (WT) and Spontaneous Hypertensive Rats (SHRs) (300-380 g) received i.p. injections of Vehicle (NaCl 0.9%), Diazepam (2 mg/kg), Imipramine (15 mg/kg), Bj-PRO-7a (71, 213 or 426 nmol/kg), Pirenzepine (852 nmol/kg), α-metil-DL-tirosina (200 mg/kg) or Chlorpromazine (2 mg/kg) and were placed in the Elevated Plus Maze (EPM), Open Field (OF) and Forced Swimming (FS) tests. The heptapeptide promoted anxiolytic and antidepressantlike effects and increased the locomotion/exploration. These effects of Bj-PRO-7a seem to be strongly dependent on activation of M1R, catecholaminergic paths and dopaminergic receptors.
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    Alterações fisiológicas de gramíneas tropicais submetidas a diferentes níveis de irradiância
    (Universidade Federal de Goiás, 2017-03-24) Silva, Laudson Ferreira da; Castro, Tomás de Aquino Portes e; http://lattes.cnpq.br/7313401367533383; Silva, Silvana Petrofeza da; http://lattes.cnpq.br/6823998544968373; Silva, Silvana Petrofeza da; http://lattes.cnpq.br/6823998544968373; Castro, Tomás de Aquino Portes e; http://lattes.cnpq.br/7313401367533383; Melo, Hyrandir Cabral de; Moraes, Moemy Gomes de; Lopes, Francis Julio Fagundes
    This work aimed to analyze the growth and the partition of assimilates in plants such as Urochloa brizantha cv. Marandu, U. decumbens cv. Basilisk, Panicum maximum cv. Massai and Cynodon dactylon cv. Vaquero, submitted to three levels of luminosity (100, 50 and 25%). The plants were collected at regular intervals of seven days, from the 20th day after the emergency (DAE), to seventy six DAE, with a last collection of ninety DAE. The height (H), tiller number (TN), root dry matter (RDM), stem dry matter (SDM), leaf dry matter (LDM), leaf mass ratio (LMR), ratio between root dry matter and aerial part of the dry matter (RRAP), total dry matter (TDM), specific leaf area (SLA), leaf area ratio (LAR), liquid assimilation rate (LIAR), relative growth rate (RGR), leaf area index (LAI), crop growth rate (CGR), chlorophyll and carotenoids content. The analyzed species grew and developed in an environment with 50% of light equivalent to plants grown in full sun, since there were no significant differences regarding H, RDM, SDM, LDMC, TDM. Similarly, there was no significant difference in LIAR, RGR and CGR in the species, except in P. maximum, which presented such physiological indicators of growth higher than plants grown in the full sun. LAI was also equivalent, except for U. brizantha and U. decumbens, which presented higher index than plants in full sun. The luminosity level of 25% negatively affected the growth and development of the species, except in P. maximum, whose results were equivalent to those presented in full sun and 50% of light. Species demonstrate tolerance to light restriction and, therefore, production potential to be used in silvopasture and agrosilvopasture production systems.
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    Análise do secretoma de isolados do fungo Trichoderma asperellum (TR356) em resposta ao fungo fitopatogênico Sclerotinia sclerotiorum
    (Universidade Federal de Goiás, 2014-10-23) Rodrigues, Amanda Rafaela; Ulhoa, Cirano José; http://lattes.cnpq.br/8368469162867277; Ulhoa, Cirano José; Monteiro, Valdirene Neves; Fernandes, Kátia Flavia
    The genus Trichoderma is represented by non-pathogenic soil fungi that have been studied by act as biological control agents against fungal pathogens, such as Sclerotinia sclerotiorum, the fungus that causes white mold, besides that, reduce environment and human health risks by being able to replace agrochemicals. Proteomic strategies with MS techniques have been important tools in studies of patterns identification for protein expression in different growth conditions. This work aims to development new strategies that enable the detection and identification of proteins secreted by Trichoderma asperellum (TR356) and Sclerotinia sclerotiorum when inoculated together and separated. The results obtained by MALDI / TOF analysis allowed the identification of a β-1.3--glucanosiltransferase and α-1.2-Dmannosidase, demonstrating the possibility of proteins identification for better comprehension about interaction between these organisms. It was therefore possible to identify the proteins through strategies used, but further analysis are required in order to elucidate the function and interaction of proteins secreted by the fungus Trichoderma asperellum TR356 against S. sclerotiorum.
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    Análise da dinâmica do glicoproteoma de trichoderma asperelloides durante o micoparasitismo
    (Universidade Federal de Goiás, 2022-01-21) Naoum, Stéphanie; Monteiro, Valdirene Neves; http://lattes.cnpq.br/8264822485508916; Ulhoa, Cirano José; http://lattes.cnpq.br/8368469162867277; Ulhoa, Cirano José; Bailão, Alexandre Melo; Georg, Raphaela de Castro; Paula, Renato Graciano de
    Glycosylation is a post-translational modification that occurs in most cells and is an important mechanism for several cellular processes such as protein secretion, cell signaling, protein translocation and stability, maintenance of cell structure and receptor-ligand interactions. In fungi there is a set of enzymes specialized in glycosylation of proteins, exerting functions related to the structure of the cell wall and the cell as a whole, assisting in integrity, growth, differentiation and signaling. Fungi of the genus Trichoderma are known for their ability to biocontrol through mycoparasitism mechanisms involving the production of cell wall degrading hydrolytic enzymes. Therefore, the objective of this work was to identify the N-glycosylated proteins produced by T. asperelloides (TR356) during mycoparasitism. The Concanavalin A (ConA) affinity chromatography technique was used to enrich the samples and select N-glycated glycoproteins with oligomannosidic structure. In the interaction between the fungi, the contact condition showed a difference in the protein content when compared to the control samples. The specific activities of the enzymes β-1,3-exoglycanase, β-1,3-endoglycanase, chitinase, N-acetyl-glucosaminidase, β-glucosidase, acid phosphatase, α-mannosidase, α-arabinofuranosidase and demonstrated a significant increase in activities in conditions before contact and contact. Several proteins were identified in the samples using the mass spectrometry technique. A total of 253 proteins were identified in the control sample. In samples referring to before contact and contact between T. asperelloides (TR 356) and S. sclerotiorum the number of identified proteins was higher, 582 and 524 proteins, respectively. We can infer that the presence of the pathogen S. sclerotiorum in the same environment as T. asperelloides stimulates the production of specific proteins for this situation, necessary for mycoparasitism. Glycoproteins with different amounts of N-glycosylation sites involved in mycoparasitism were identified, and the number of glycoproteins and N-glycosylation sites increased in pre-contact and in-contact situations. Most secreted proteins are involved in carbohydrate metabolism and transport, cell signaling, and post-translational modifications and folding. The identified proteins of the intracellular environment are involved in post-translational modification and protein folding, in carbohydrate metabolism and transport, and in cellular metabolism in general. Finally, we observed that a significant number of identified proteins still do not have a defined function, which can be considered an important source of new studies and new knowledge in relation to mycoparasitism.
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    Análise da diversidade genética em populações de Sclerotinia sclerotiorum
    (Universidade Federal de Goiás, 2010-08-31) Nascimento, Lucas Breseghelo do; Silva, Silvana Petrofeza da; http://lattes.cnpq.br/6823998544968373
    The fungus Sclerotinia sclerotiorum is among the most successful pathogens, omnivores, and without specific hosts, is considered one of the most important fungal pathogens in the world. Is distributed in all producing regions, temperate, subtropical or tropical. The fungus produces resistant structures called sclerotia on the surface and within tissues colonized, they returned to the soil with crop residues and are responsible for the survival of the fungus in the same area for up to eight years. The objectives of this study were to quantify the variability within and between populations of the fungus S. sclerotiorum in bean and soybean crops in different producing places of those varieties. 46 isolates were collected of S. sclerotiorum in six locals of grain production in different regions of Brazil. The sites chosen were Monte Carmelo-MG, Formosa-GO, Lucas do Rio Verde-MT, Montividiu-GO, Londrina, PR and Santo Antonio de Goiás-GO. All areas of the original host of the gathering was the bean with the exception of Londrina-PR in which the host was soybeans. A population study of the fungus through RAPD markers using 13 primers was carried out for the analysis of genetic variability of the fungus. In parallel, tests were also made for the the Mycelial compatibility groups among isolates and test production of hydrolytic enzymes. The statistical analysis was performed using the Arlequin software, which indicated variability among populations of 16.94% and 83.06% within populations. Were found 10 mycelial compatibility groups without specific populations .Enzyme activities performed indicated significant differences within the populations of all enzymes, a comparison between populations also showed significant differences among populations in polygalacturonases, 1,3-β-glucanase and xylanase. The results indicate a high level of variability within populations and low variability among populations.
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    Análise da expressão de genes envolvidos na manutenção da homeostase de cobre no patógeno humano Paracoccidioides brasiliensis
    (Universidade Federal de Goiás, 2009-05-29) SANTOS, Rodrigo da Silva; SOARES, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637
    The fungus Paracoccidioides brasiliensis is a human pathogen with a wide distribution in Latin America. The fungus causes paracoccidioidomycosis when mycelia reachs the lungs. The success of the infection depends on the acquisition of essential micronutrients such as copper, which is required as cofactor for a variety of enzymes important in biological in several processes, such as respiration, growth and acquisition of iron. Previous studies, of the Laboratory of Molecular Biology showed that a high affinity copper transporter (PbCTR3) is a molecule highly expressed and probably necessary for the infection establishment of by P. brasiliensis. In the present study were isolated and characterized the genomic and cDNA sequences encoding for PbCTR3 of P. brasiliensis. The cDNA presented 582 base pairs and encodes for a protein with 193 amino acids, predicted molecular mass of 21.5 kDa and pI of 8.6. The genomic sequence has four exons interrupted by three introns. In silico analysis was performed on the database of the structural genome of P. brasiliensis (http://www.broad.mit.edu/annotation/genome/paracoccidioides_brasiliensisMultiHome.html) , where genes involved in maintaining the homeostasis of copper have been identified and used to design of a model of copper homeostasis in P. brasiliensis. The transcriptional behavior of Pbctr3 and genes involved in copper homeostasis were examined during exposure of yeast cells of P. brasiliensis to copper and iron depletion conditions, by real time qRTPCR. It was demonstrated a significant change in transcription lovel those of genes in the absence of copper, as well as in the combined absence of both metals. qRT-PCR was used to analyze the expression of Pbctr3 and Pbcrp (which encodes a protein responsive to copper) in yeast cells of P. brasiliensis derived from infected lungs and spleen at different time points of infection. The expression of Pbctr3 and Pbcrp was super-regulated during experimental infection. Taken together, these data suggest the importance of PbCtr3 and of absorbing copper / iron during the infectious process
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    Análise da influência dos polimorfismos 1236C>T, 2677G>T/A E 3435C>T do gene ABCB1 na resposta ao tratamento com clozapina
    (Universidade Federal de Goiás, 2015-12-04) Salamanca, Ayda Luz Malaver; Santos, Rodrigo da Silva; Ghedini, Paulo César; http://lattes.cnpq.br/5789550234984454; Ghedini, Paulo César; Brito, Rodrigo Bernini; Bicudo, Lucilene Arilho Ribeiro; Santos, Rodrigo da Silva
    Clozapine (CLZ) is the antipsychotic drug of choice in treatment refractory schizophrenia (TRS), however 30% of the patients with TRS don’t have full response to treatment with CLZ, these patients are considered to have super refractory schizophrenia (SRS). The response variability to treatment with CLZ may be associated with alterations of the CLZ plasma levels promoted by 1236C>T, 2677G>T/A e 3435C>T ABCB1 gene polymorphisms. ABCB1 is a transmembrane glycoprotein of transport, which export drugs from the intracellular to the extracellular space. ABCB1 polymorphisms cause structurally and functional protein changes that influence the intracellular CLZ levels and, consequently, promoting therapy failure. This work had the aim to establish the relation between the 1236C>T, 2677G>T/A e 3435C>T ABCB1 polymorphisms and the CLZ treatment response. This study included a total of 68 patients, 34 of whom were classified as TRS (CLZ responders) and 34 as SRS (CLZ non-responders). All patients were in use of CLZ for at least one year. For the genotype test was extracted genomic DNA, following for PCR and sequencing techniques. It was not observed any differences in allelic and genotype distribution between 1236C>T, 2677G>T/A and 3435C>T polymorphisms between RS and SRS groups. The polymorphisms not influenced the CLZ dose level, and percentage of BPRS change. In the same way, the results not showed relation between age, coffee intake, smoking behaviors and ethnicity and the CLZ treatment response, however, was evidenced a higher proportion of female patients in the SRS group when compared with the TRS group. In opposite, a lower proportion of male patients was observed in the SRS than the TRS group. Taken together, the results here obtained showed no association between 12361236C>T, 2677G>T/A e 3435C>T polymorphisms of ABCB1 gene and failure to CLZ treatment.
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    Análise da tolerância á resistência em Trichoderma harzianum
    (Universidade Federal de Goiás, 2017-07-31) Santos, Karina Roterdanny Araújo dos; Georg, Raphaela de Castro; http://lattes.cnpq.br/3971276154067590; Georg, Raphaela de Castro; http://lattes.cnpq.br/3971276154067590; Campos, Ivan Torres Nicolau de; Izacc, Silvia Maria Salem; Monteiro, Valdirene Neves; Lorenzón, Esteban Nicolás
    The presence of heavy metals in the soil has increased, causing many problems of an agricultural nature. Trichoderma are a species which is among the most well know and used biocontrol agents wordwide, as well as possessing qualities such tha parasitism and antibiosis, they have great resistance to metals and can also absorb them. Therefore, it is necessary for more detailed studies of the resistance of this fungus to metals and to research further the processes involved. In this work, we aimed to better understand the tolerance of T. harzianum to aluminum chloride, though a series of experiments performed in BDA, MEX e MYG, as well Bradford tests for quantification of proteins. In addition, we evaluated the gene response to metal stress through the sequencing RNA samples obtained when the fungus was created in BDA medium and submitted to concentrations of 1.5 and 3.0 mg / mL of aluminum chloride, 1.5 mg / mL, the fungus showed significant inhibition. It was observed that T. harzianum showed higher mycelial growth when grown in BDA medium compared to MEX and MYG, presenting levels of protein secretion inversely proportional to increasing concentrations of aluminum. The amount of transcriptionspecific factors of the stress response was increased, as well as the induction of genes involved in G-mediated cell signaling and increase in genes involved in vacuolar transport and carrier proteins. A repression of genes encoding proteins associated with cellular processes important for the growth of T. harzianum was also observed. These results show that T. harzianum is resistant to aluminum since it is able to tolerate and absorb large concentrations of this metal, which is commonly harmful to agriculture.
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    Análise das proteínas de reserva do arroz silvestre oryza glumaepatula e de linhagens interespecíficas oryza sativa x o. Glumaepatula
    (Universidade Federal de Goiás, 2010-03-31) SANTOS, Karina Freire D'eça Nogueira; DIDONET, Claudia Cristina Garcia Martin; http://lattes.cnpq.br/2890489628230874; BRONDANI, Claudio; http://lattes.cnpq.br/4775600104554147
    The objective of this dissertation was to analyze quantitative and qualitatively the total storage protein content and its fractions albumin, globulin, prolamin and glutelin from the grain of 29 genotypes of the wild rice Oryza glumaepatula and 70 interspecific lines backcross 2 obtained from the cross O. sativa x O. glumaepatula. From these lines, 34 BC2F8 were obtained from the cross O. sativa BG 90-2 x O. glumaepatula RS-16 and 36 BC2F10 were obtained from the cross O. sativa CICA-8 x O. glumaepatula RS-16. From the analysis of variance for the total protein content and its fractions, it were found highly significant differences (P<0.01) between the wild genotypes and the interspecific lines. The average of total protein content of wild genotypes was 12.17%, whereas the lines of BG 90-2 x RS-16 showed an average of 7.05% and the lines of CICA-8 x RS-16 showed an average of 8.40%. The wild genotype BGA14280 showed the highest total protein content (14.94%). In the comparative analysis of interspecific lines and their parents, it were found five lines with higher total protein content (average of 10.95%), which was significantly superior to the cultivated parent BG 90-2 (10.0%) and CICA-8 (9.61%). However, these lines showed lower content in relation to the wild parent RS-16 (14.06%). In relation to the protein fractions, 40 interspecific lines showed higher values in relation to their parents, excepting in one occasion, where the wild parent RS-16 showed higher value to the glutelin fraction. Considering the 29 wild genotypes, it were found the highest contents of albumin, prolamin and glutelin, excepting to the globulin fraction, where the wild genotype was not significantly different from the cultivar BRS Bonança. The SDS-PAGE analyses of the total protein and the glutelin fraction showed a differential profile of α-glutelins for the wild genotypes, emphasizing the BGA14232 genotype, which did not showed the α-polypeptides commonly identified in the remaining genotypes. Considering the interspecific lines and their parents, it was found similar profile of total protein and α-globulins, with differences in relation to the 40 kDa α-glutelin, which was found just in the wild parent RS-16, indicating a differential expression in O. glumaepatula, since the O. sativa showed a 39 kDa α-glutelin. In relation to albumin, globulin and prolamin fractions, the wild genotypes showed a different protein profile in comparison to the interspecific lines and the cultivated rice, probably due to the two crosses in direction to BG 90-2 and CICA-8, which were the recurrent parents during the interspecific lines development. The highest protein content found to the O. glumaepatula, the different protein profile and the finding of interspecific lines with higher protein content in relation to their cultivated parents clearly show the positive contribution of this species to the genetic breeding aiming the increase of the nutritional value for the grain of the cultivated rice in Brazil.
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    Ánálise de alterações no gene receptor de andrógeno em homens com infertilidade idiopática
    (Universidade Federal de Goiás, 2009-03-31) MESQUITA, Wyara Elanne de Jesus Castro; JESUÍNO, Rosália Santos Amorim; http://lattes.cnpq.br/5113656623817587
    Male idiopathic infertility is related to defects in normal spermatogenesis, due to genetic causes. The spermatogenesis is a dependent process on high levels of male sex hormones, the androgens. The androgen, in turn, perform its function when associated with the androgen receptor (AR), protein encoded by AR gene. Mutation in AR gene lead to a synthesis of non functional AR, which results in the failure of the process of spermatogenesis and, consequently, causes male infertility. This work has as its main objective the verification of the occurrence of mutation in the AR gene in patients with male idiopathic infertility who come from the HC-UFG Human Reproduction Center. Samples were analyzed from 206 patients. The result was that 95 patients were found to be normal while 111 with an altered result for the spermogram. The samples were amplified for exons 1, 4, 6, 7 and 8 of the AR gene and the results subjected to statistical analysis, Mann Whitney, logistic regression, and chi tests. The existence of the relationship between defects of sperm and AR gene mutation was verified. The analysis of the relationship between the spermogram and the AR gene mutation in five evaluated exons was significant only for exons 1 and 7. Patients with numerical unsettled spermogram had a higher frequency of mutations in exon 7, teratozoospermics in exon 1 and exon 7 in astenozoospermics patients. Exons 4, 6 and 8 showed no meaningful statistical relationship in reference to the alteration of the spermogram. Among results related to social custom, alcohol proved to be significant for mutation in the AR gene. This study has reaffirmed the relationship between the presence of mutation in AR genes as probable causes of defects in spermatogenesis. Consequently, male idiopathic infertility depends not only on the genetic factor, but also on the association between this factor and the environment where man inhabits
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    Análise de diferentes misturas enzimáticas do fungo Humicola grisea var. thermoidea na hidrólise da fração de hemicelulose de bagaço de cana-de-açúcar
    (Universidade Federal de Goiás, 2011-08-31) Faria, Syd Pereira; Castro, Carlos Henrique de; http://lattes.cnpq.br/6354834854727314; Faria, Fabrícia Paula de; http://lattes.cnpq.br/3739169267521003; Faria, Fabrícia Paula de; Lopes, Francis Júlio Fagundes; Ulhoa, Cirano José
    The biomass consists mainly of cellulose, hemicellulose and lignin. Cellulose is the most abundant polymer xylan and the main component of hemicellulose. Enzymatic hydrolysis is an important step in the bioconversion of cellulose and hemicellulose fraction of lignocellulosic residues. However, for the conversion of the hemicellulose, the substrate must undergo a pretreatment step which allows the refining of biomass at the microscopic level, separating the same into its main components (cellulose, hemicellulose and lignin) or loosening the fibers of its components, facilitating the accessibility of enzymes to hemicellulose chain. The conversion of cellulose and xylan to glucose and xylose may be performed by a group of enzymes produced by bacteria and fungi. The thermophilic fungus Humicola grisea var thermoidea produces an efficient complex of cellulolytic enzymes (endoglucanases, cellobiohydrolases and β-glucosidase) and xylanolítics (endoxylanases and β-xylosidase) with high thermal stability when cultivated in different lignocellulosic substrates. The aim of this study was to analyze the efficiency of enzymes produced by H.grisea in the hydrolysis of the hemicellulose fraction of sugarcane bagasse (BCA). BCA assays used in the hydrolysis was subjected to alkaline pretreatment (Treatment I, II and III) and steam explosion. BCA pretreated was subsequently hydrolyzed with different enzyme mixtures using culture supernatant of the fungus H. grisea supplemented with recombinant enzymes endoxylanase (HXYN2r), cellobiohydrolase (CBH1.2r) and β-xylosidase (XYNB2r). The results showed that the pre-treatment was most suitable to steam explosion which revealed a yield of about 78% xylan. The best enzyme formulation was a mixture of the supernatant of culture of H. grisea (SHG) grown in wheat bran (WB) and BCA supplemented with HXYN2r (950 U), CBH1.2r (950 U) and XYNB2r (1.17 U).
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    Análise de populações de Sclerotinia sclerotiorum em cultura de feijoeiro através de marcadores SSR.
    (Universidade Federal de Goiás, 2009-09-30) GOMES, Eriston Vieira; SILVA, Silvana Petrofeza da; http://lattes.cnpq.br/6823998544968373
    79 Sclerotinia sclerotiorum isolates were collected under central pivot irrigations sistems, from Brazilian fields, divided in 4 populations (S2, S3, A and M) and analyzed to determine the genetic variability among and between populations using molecular markers based on microsatelittes. 10 primers were used and the amplification products were separate in poliacrilamida gels and the bands were silver stained. A total of 102 different haplotypes were identified, and the amount of haplotypes varied from 6 to 18 for locus. The genotypic diversity ranged from 65% to 91%. Analyses based on genetic diversity and fixation indices which indicate the variability between populations was 28.79% (FST = 28793) and the variability among populations was 71.21%. The Jaccard similarity index indicated that the populations S2 and A is genetically closer. The population S3 presented a similarity index of 0.44 compared with the populations S2 and A. The population M, originated from several collection sites, was considered genetically more distant showing a index of 0.46 compared with the others populations . The high variability between and among populations can indicate that, besides the possible introduction of new genotypes in the analyzed fields, could be having clonal and sexual reproduction in isolated of S. sclerotiorum from Brazilian cerrado.
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    Análise de transcritoma e expressão gênica de feijoeiro comum (Phaseolus vulgaris L.) submetido ao estresse abiótico
    (Universidade Federal de Goiás, 2016-03-08) Pereira, Wendell Jacinto; Vianello, Rosana Pereira; http://lattes.cnpq.br/4523999698824309; Lanna, Anna Cristina; Brondani, Claudio; Vianello, Rosana Pereira
    The common bean (Phaseolus vulgaris L.) is an important foodstuff strongly associated with an issue if global food security. Among the factors that affect their agricultural production, abiotic stress episodes listed as high risk and impact, can restrict the area under cultivation and crop yields. The characterization of high-throughput differential gene expression involved in plant response to abiotic stresses enables profile the genes, and the metabolic pathways, involved in the mechanisms of tolerance to abiotic stresses. In this study, the expression of genes related to response to drought stress in leaf and root tissue of two contrasting common bean genotypes for stress tolerance was evaluated by RNA-Seq. A new Phaseolus vulgaris transcriptome was built featuring 1,668 new loci and 23,169 new isoforms. In all, 54,807 transcripts were identified, found being distributed in 28,640 genes. Through the edgeR package (Bioconductor), they determined by differential expression analysis of leaf and root 1,242 differentially expressed genes. Of these, 458 genes had different expression in tolerant genotype (191 up-regulates and 267 down-regulated) when compared to the sensitive genotype. Functional annotation revealed in tolerant genotype predominance of genes in the categories enriched oxidoreductase activity, oxidation-reduction process, regulation of gene expression, regulation of macromolecule metabolic process and dioxygenase activity. By qPCR, of the 15 DEGs identified via RNA-Seq and selected for characterization, 74.42% were validated for differential expression. Additionally, a total of 151,283 variants were identified, of which 135,167 are SNPs and 16,115 are indels. Through functional annotation, performed in SnpEff v.4.2, were identified 330,378 effects caused by variants. The data used in this study were deposited in the Sequence Read Archive (NCBI) and will be available. The results of this study contribute to the expansion of knowledge about the gene mechanisms, as well as their functional variants, related to abiotic stresses tolerance in beans.