Programa de Pós-Graduação em Biotecnologia - PPGBIOTEC/ICB
URI Permanente desta comunidadehttps://repositorio.ufpa.br/handle/2011/6089
O Programa de Pós-Graduação em Biotecnologia (PPGBIOTEC) é um programa do Instituto de Ciências Biológicas (ICB) da Universidade Federal do Pará (UFPA). O PPGBIOTEC fornece a base teórica para o desenvolvimento de projetos em duas áreas de concentração e quatro linhas de pesquisa, definidas de acordo com a competência e experiência de orientação dos docentes do programa: Área de Bioprospecção: linhas de pesquisa: (i) química orgânica de biomoléculas e metabolômica; e (ii) genômica, transcriptômica, proteômica e bioinformática de microrganismos e plantas (Bactéria, Archaea e Eukarya). Área de Biotecnologia Aplicada: linhas de pesquisa: (i) saúde humana e veterinária; e (ii) produção de insumos e processos industriais.
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Item Acesso aberto (Open Access) AutoAssemblyD software para submissão e gerenciamento de montagem de genomas a partir de modelos XML(Universidade Federal do Pará, 2014-01-24) VERAS, Adonney Allan de Oliveira; SILVA, Artur Luiz da Costa da; http://lattes.cnpq.br/7642043789034070Technologies for second-generation sequencing provided a major breakthrough of the genome, making its use a landmark that has revolutionized biology. These platforms are characterized by a reduction in sequencing time, high data production and low cost per base sequenced, however, these devices produce data mostly consist of short readings which represents a major challenge for reconstruction of the genome due to this new feature readings of computational tools had to be developed to accomplish the task of assembling their example we Velvet, AllPaths, Abyss, SOAPdenovo2, Edena. However, most of these applications are executed through command lines extended and composed of several parameters must follow the standard syntax to use, because in case of errors in the syntax is the possibility of not obtaining the best result, with the aim of solve this problem we present the AutoAssemblyD that besides providing the use of these assemblers through a graphical interface also enables the management of these executions remotely.Item Acesso aberto (Open Access) Caracterização antigênica, físico-química e biológica do vírus BE AR 701405, obtido a partir de mosquitos da espécie Psorophora (Jan) ferox, capturados em Altamira – Pará(Universidade Federal do Pará, 2014-03-26) ARAÚJO, João Batista dos Santos; LEAL, Élcio de Souza; http://lattes.cnpq.br/1158983666415285The BE AR 701405 virus was isolated from a pool of Psorophora (Jan.) ferox mosquitoes captured in the municipality of Altamira, Para state, Northern Brazil, in 2006. The objective of this study was the physicochemical, biological and antigenic characterization of the BE AR 701405 virus in order to provide data for it taxonomic classification. Newborn mice inoculated by intracerebral route showed evidences of neurological manifestations, such as chill and motor disorders after the infection with the virus BE AR 701405. Cytophatic effect (CPE) was in infected Aedes albopictus C6/36 cells was not clearly observed, however in VERO cells CPE was observed after 48 hors post infection. The virus load was determined in 10-4,1 LD50/ 0.02 mL and the titer after DCE analysis was calculated in 10−2,6 LD50/0,02 mL. The BE AR 701405 virus reacted antigenically with Pixuna virus hiperimmune serum. In conclusion, the studied vírus was sensible to the DCA solvent suggesting that it is an enveloped virus. Furthermore, by serology this virus was identified as a member of the group A, genus Alphavirus, family Togaviridae, more closely related to Pixuna virus. In addition, newborn mice, as well as C6/36 and VERO cells demonstrated to be sensible to the infection by the BE AR 701405 vírus. Further studies are needed to better understand the antigenic relationship between the BE AR 701405 virus and Pixuna virus.Item Acesso aberto (Open Access) Prospeção de aminoácidos tipo micosporinas (MAAs) em cianobactérias da Amazônia oriental(Universidade Federal do Pará, 2014-06-30) ARAÚJO, Sanclayver Corrêa; GONÇALVES, Evonnildo Costa; http://lattes.cnpq.br/8652560763793265Mycosporine-like amino acids (MAAs) are low molecular weight water soluble compounds wuich biosynthesis was thought to occur via shikimate pathway in bacteria, phytoplankton and macroalgae. Recently evidences show its biosynthesis via the pentose pathway in a cyanobacterium. Due to its high extinction molar coefficient in UV region, they provide protection against the damaging effects of UV radiation and have potential for use in sunscreens. In previous works it was described a biosynthetic cluster for shinorine with the enzymes dehydroquinate synthase, present in the shikimate pathway, O-methyltransferase, and ATP-grasp and NRPS-like enzymes. It was observed that the enzyme 2-epi-5-epi-valiolone synthase, which link the pentose pathway with MAAs production would lead to formation of 4-deoxygadusol, precursor of MAAs. In this work a genomics approach was carried out so find these genes so that MAAs production potential in Amazonian cyanobacteria of the Coleção Amazônica de Cianobactérias e Microalgas, CACIAM 14, CACIAM 53, CACIAM 54 and CACIAM 57 strains was assessed, as well as an analysis through capillary chromatography coupled to electrospray mass spectrometryof the metabolites in CACIAM 14. The genes Dehydroquinate synthase and o-methyltransferase were found in all the strains studied. The genes of pentose pathway Ribulose-phosphate-3-epimerase and transketolase were detected in the strains CACIAM 14, CACIAM 53 and CACIAM 57, what may suggest that in these strains MAAs could be produced by both pathways while in CACIAM 57 MAAs could be produced by shikimate pathway only. In CACIAM 14 strain, fragments of mass to charge relationship 186 and 197 were found in the same retention time of a precursor ion of m/z 333, what could be related to shinorine. Therefore, the strains CACIAM 53, 54 and 57 are potentially producers of MAAs due to possessing biosynthetic genes and CACIAM 14 is shinorine producer.Item Acesso aberto (Open Access) Co-cultivo de embriões bovinos com células-tronco adultas derivadas de tecido adiposo(Universidade Federal do Pará, 2014-09-05) NASCIMENTO, Hamilton Silva do; MIRANDA, Moysés dos Santos; http://lattes.cnpq.br/3354029928888919The in vitro embryo production (IVP) is a biotechnology used to increase the reproductive potential of genetically superior animals, embryos produced in vitro are inferior in quality when compared in vivo. Techniques trying improve the rates of in vitro produced embryos. One technique is the co-culture system using somatic cells that remove toxic metabolites and protect against oxidative stress. Mesenchymal stem cells derived from adipose tissue (CTA) are multipotent cells that secrete growth factors and cytokines. The stem cells were used in in vitro co-culture of bovine embryos at different concentrations in order to improve the protocol PIVE. CTAs underwent differentiation into three mesenchymal lineages, and was performed immunophenotyping of specific membrane markers of MSCs. The cleavage rate was assessed on the second day after fertilization and blastocyst rate on the seventh day when they were stored for counting the total number of cells and gene expression. The results were analyzed by ANOVA, T-test and post-test of Fisher, adopting a significance level of 5%. Treatment of co-cultivation with CTAs significantly influenced the formation of the blastocyst, the total number of cells in embryos and correlated gene expression pluripotency and carbohydrate metabolism. These results showed increased production rate and quality of embryos produced in vitro co-culture with CTAs compared to co-culture with granulosa cells. These results also indicate that the constant presence of CTAs in co-culture is superior to conditioning with CTAs. Checked the effects of CTAs can occur through soluble factors or via exosomes secreted by CTAs. Future studies are needed to clarify the possible cause for positive effects observed in this work by the CTAs in co-cultivation.Item Acesso aberto (Open Access) Influência dos metabólitos secundários de Piper divaricatum da região amazônica no controle do Fusarium solani f. sp. piperis causador da fusariose em pimenta do reino(Universidade Federal do Pará, 2014-09-18) MEIRELES, Erisléia das Neves; RAMOS, Alessandra de Rezende; http://lattes.cnpq.br/1279694874191138; SILVA, Joyce Kelly do Rosário da; http://lattes.cnpq.br/2278686174214080The culture of black pepper (Piper nigrum L.) is one of the main agricultural activities in Pará State and it suffers serious damages caused by fusariosis, disease restrict in Brazil. This paper evaluated the antifungal activity in vitro of essential oils of species of Piper rich in phenylpropanoid before the Fusarium solani f. sp. piperis, which causes the fusariosis. The inhibition of mycelial growth by the method of agar dissemination in concentration of 5 mg.mL-1 was considered: decrease to P. aduncum (20.3%) and P. Krukoffii (31.4%); moderated to P. callosum (55.7%) and P. marginatum (70.3%) and high to P. divaricatum (93.3%). The major components identified by CG-EM were dilapiol (92.0%), safrole (78.0%), methyleugenol (75.2%) and eugenol (7.9%), apiol (80.0%), Z-isoosmorizol (44.0%) and E-anethole (22.0%), respectively. The oil of P. divaricatum and its major compounds present CIM values of 0.75 mg.mL-1. The evaluation of the effects combined of eugenol and methyl eugenol pointed eugenol as the main responsible by the activity.Item Acesso aberto (Open Access) Acúmulo de colesterol por Mycobacterium smegmatis como possível modulador da biossíntese de lipomanana e lipoarabinomanana(Universidade Federal do Pará, 2015-02-26) SANTOS, Ana Cristina Doria dos; SENA, Chubert Bernardo Castro de; http://lattes.cnpq.br/8620752020290438Mycobacterial cell wall is a hallmark of Mycobacterium genus, constituted of bioactive lipids and glycoconjugates: phosphatidylinositol mannoside (PIMs), lipomannan (LM) and lipoarabinomannan (LAM). The chronic infection inside lung macrophages is related with cholesterol accumulation into host cells as an alternative source of carbon and energy to maintain the bacilli with its physiological functions. To understand the activity of these immunomodulatory glycoconjugates during adaptation under the unusual environment inside the host infected cells, the present work propose to investigate the possible modulation of LM/LAM biosynthesis in Mycobacterium smegmatis (saprophytic) after culture in minimal medium (MM), supplemented with glycerol and/or cholesterol. Our results showed that saprophytic bacilli are able to accumulate the cholesterol and change the bacterial physiology due to slow growth and restrict cell density. Furthermore, the cholesterol consumption decreased the accumulation of PIMs and promoted morphological changes and bacterial aggregates, even maintaining the cell wall with its specific physic-chemistry characteristic (alcohol-acid resistance). The most impressive change after cholesterol consumption was the LAM biosynthesis, which showed distinct electrophoresis migration, compatible to high molecular weight of LAM from non-saprophytic bacilli (from 25 – 30 KDa to 30 – 50 KDa). These results showed that cholesterol consumption, when utilized as principal alternative of carbon and energy source, is able to induce physiological changes in mycobacteria, mainly in LAM biosynthesis, one of the most immunoregulatory molecules of cell wall. Ours data suggest that similar changes in mycobacteria may occur inside the granuloma, and that changes may help the evolution of tuberculosis to chronic and multibacillary form, hallmarked by immunodeficient aspect against the bacilli.Item Acesso aberto (Open Access) O papel do colesterol na biossíntese da parede celular de Mycobacterium smegmatis(Universidade Federal do Pará, 2015-02-26) MARINHO, Victor Hugo de Souza; SENA, Chubert Bernardo Castro de; http://lattes.cnpq.br/8620752020290438Different Mycobacterium species are causative agents of disease in humans, for example, the tuberculosis. All mycobacteria have a complex cell wall, distinct of others bacteria, conferring specific physic-chemical characteristic to Mycobacterium genus, due to protect against immune system and waterproofing against the intake of much antibiotics. During infection, the bacillus is able to adapter to harsh environment, due to consumption of cholesterol from itself host cell (macrophages) as alternative carbon and energy source. That nutritional aspect has been considered as essential for division of bacilli and consecutive progress of tuberculosis disease. The present study has as objective to evaluate in vitro the modulation of saprophytic Mycobacterium smegmatis cell wall biosynthesis after cholesterol consumption as primordial energy and carbon source. As results, we are found by Thin Layer Chromatography (TLC) that bacillary adaptation to microenvironment with poor nutrient (minimal media – MM) maintained the biosynthesis and accumulation of essential cell wall components, when the growth occurs in presence of someone defined carbon and energy source (glycerol and/or cholesterol). Among them without changes, we analyzed Trehalose Dimicolate (TDM) and the phospholipids (phosphatidylinositol (PI), phosphatidylinositol manosides (PIMs), Cardiolipin (CL) and phosphatidylethanolamine (PE)). Differently of these results, the micolic acid showed representative accumulation, comparing with 7H9 culture, only when the MM was supplemented with glycerol. This result was confirmed by alcohol-acid staining using fluorescent auroamine dye, suggesting some changes in physic-chemistry cell wall properties. On the other hands, the MM culture induced the accumulation of glycopeptidolipids (GPLs), independently of glycerol/cholesterol supplementation. Such disturbance in cell wall biosynthesis also changed the bacillary hydrophobicity in all MM groups, but does not change the resistance and sensibility to antibiotics. Those results clearly show that cell wall biosynthesis might be modulate during nutritional shortage, and such presence or absence of cholesterol, as occurs during infection, does not significantly change the bacillary physiology to become vulnerable for antibiotics. It suggests that such modulations might also occur during infection, maintaining the bacilli available to develop the tuberculosis diseases.Item Acesso aberto (Open Access) Avaliação do viés GC em plataformas de sequenciamento de nova geração(Universidade Federal do Pará, 2015-03-05) PINHEIRO, Kenny da Costa; RAMOS, Rommel Thiago Jucá; http://lattes.cnpq.br/1274395392752454The emergence of high throughput sequencing (HTS) platforms increased the amount of data making feasible to obtaining complete genomes. Despite the advantages and the throughput produced by these platforms, the high or low genomic coverage in the regions of the genome can be related to GC content. This GC bias may affect genomic analyzes and the genomic/transcriptomic analysis based on de novo and reference approach. In addition, the ways to evaluate the GC bias should be fit to data with different profiles of the GC vs coverage relationship, such as linear and quadratic. Thus, this work proposes the use of Pearson's Correlation Coefficient (r) to analyze the correlation between GC content and coverage, allowing to identify the strength of linear correlation and detect nonlinear associations, beyond identify a relationship between GC bias and sequencing platforms. The positive and negative signs of r also allow us to infer directly and inversely proportional relationships, respectively. To evaluate the bias, we used the data of Corynebacterium pseudotuberculosis obtained from different sequencing technologies to identify if the CG bias is related to used platforms.Item Acesso aberto (Open Access) Busca e classificação sistemática das proteínas oxigenases com ferro não hêmico em plantas(Universidade Federal do Pará, 2015-03-06) SOUSA, Kellen Rayanne Matos de; DARNET, Sylvain Henri; http://lattes.cnpq.br/4586614214029929Nonheme iron oxygenase proteins shares a conserved domain consisting of eight histidines, and they can be found in eukaryotes and prokaryotes organisms and participate in important pathways of lipid biosynthesis. To understand the evolutionary relationship among these proteins, we performed comparative and phylogenetic analyzes in prokaryotes and eukaryotes that allowed a classification of this family, nonexistent until now. The search of sequences resulted in a collection of 448 proteins, belonging to 58 organisms previously selected. The multiple alignment made with MAFFT (BLOSUM 62; L-INS-i) showed the presence of three histidine-rich motifs, with conserved spacing among them. The classification made with CLANS software has generated 28 clusters through of similarity among sequences. Two clusters contain sequences that had no similarity to proteins already characterized, and 48 sequences were not assigned to any of the 28 clusters. In the collection, 119 sequences are derived from plants, distributed in 7 clusters corresponding to C4 methysterol monooxygenase, C5 sterol desaturase, fatty acid hydroxylase, sphingolipid C4 monooxygenase, aldehyde decarbonylase, β-carotene hydroxylase and Acyl-ACP desaturase functions. Phylogenetic analysis using maximum likelihood method with PhyML tool showed the formation of well-defined groups that were similar to generated by CLANS. These results start to fill a gap existing so far about the evolutionary relationship and classification of nonheme iron oxygenases. Also, suggest that within this family there are still proteins with unknown functions, reinforcing the need for more studies of functional characterization.Item Acesso aberto (Open Access) Estudo de genômica comparativa de corynebacterium pseudotuberculosis linhagem 226 (biovar ovis)(Universidade Federal do Pará, 2015-03-10) DIAS, Larissa Maranhão; CARNEIRO, Adriana Ribeiro; http://lattes.cnpq.br/7533716053525477; http://lattes.cnpq.br/7533716053525477Corynebacterium pseudotuberculosis is a gram-positive, facultative intracellular, non-sporulating and non-encapsulated bacterium, it is non-motile although it has fimbriae, and can assume coccoid or filamentous forms (pleomorphic). Its optimum growth temperature is 37°C. This pathogen has two biovars: ovis, which usually affects small ruminants and causes caseous lymphadenitis, and biovar equi, more common in equines, bovines, camelids and bubalines, causing ulcerative lymphangitis. Its infection can lead to carcass condemnation and reduction in wool production (in ovines and caprines), milk production and meat production and, consequently, economic losses for the agricultural industry worldwide. Currently there is no effective vaccine against those illnesses. To obtain a better understanding of these species biologically, the main objective of this work is to analyze, using comparative genomics, the strain C. pseudotuberculosis 226 biovar ovis, isolated from a caprine in California, comparing it to other strains from biovars ovis and equi. The synteny analysis revealed highly conserved gene order between strain 226 and other biovar ovis strains. Phylogenomic analyses showed that the strains I19 and 267 are, respectively, the closest and the more distant phylogenetically from strain 226. Among biovar equi strains, the one with the greater phylogenomic proximity to strain 226 was strain 1/06-A. Eight pathogenicity islands were predicted, with C. pseudotuberculosis best characterized virulence genes in literature being present in island 1. No new regions related to virulence genes could be found compared to other strains. 248 orthologous genes could be found between strains I19, 267 and 226, while 282 orthologous genes could be found between strains 258, 1/06-A and 226. Based in this study it is possible to assume that strains from biovar ovis have a little varied gene repertory and strains from biovar equi have less genes shared with strain 226, reinforcing the genetic diversity between these biovars.Item Acesso aberto (Open Access) Detecção de potenciais corticais antecipatórios em sinais de eletroencefalografia (EEG) durante a condução de carros(Universidade Federal do Pará, 2015-03-16) SANTOS, Fredson Carmo dos; CARVALHO, Schubert Ribeiro de; http://lattes.cnpq.br/1496976331707751; GOMES, Bruno Duarte; http://lattes.cnpq.br/4932238030330851The recognition of the driver’s intention from electroencephalographic signals (EEG) may be useful in the development of brain computer interface (BCI) to be used in synergy with intelligent vehicles. This can be beneficial to improve the quality of interaction between the driver and the car, for example, providing a response from the smart car aligned with the intention of the driver. In this study, the anticipation is considered as the cognitive state that leads to specific actions while driving a car. Therefore, we propose to investigate the presence of anticipatory patterns in EEG signals while driving vehicles to determine two specific actions (1) left and (2) turn right, a few milliseconds before such actions take place. An experimental protocol was proposed to record EEG signals of 5 individuals as they operate a virtual reality simulator non-invasive - it was designed for this experiment - which simulates driving a virtual car. The experimental protocol is a variant of the paradigm of contingent negative variation (CNV) with Go and Nogo conditions in virtual reality training system. The results of this study indicate the presence of anticipatory patterns observed in slow cortical potentials in the time domain (medium EEG signal) and the frequency (Power Spectra and phase coherence). This opens a range of possibilities in the development of BCI systems - based on anticipatory signals - that connect the driver to the intelligent vehicle favoring a decision-making to assess the intentions of drivers may eventually prevent accidents while driving.Item Acesso aberto (Open Access) Biodiversidade bacteriana durante a fermentação espontânea de frutos de açaí (Euterpe oleracea)(Universidade Federal do Pará, 2015-06-22) MOURA, Fábio Gomes; ROGEZ, Hervé Louis Ghislain; http://lattes.cnpq.br/5202118426597590Açai (Euterpe oleracea) fruits (AF) and a wide variety of products derived from this fruit are an example of success of a local and typically Brazilian product on the global market. The AF suffer spontaneous fermentation during postharvest due mainly to its high microbial load and the favourable intrinsic and extrinsic parameters. The biodiversity of microbial community in AF and their evolution between three geographical origins and two fermentation conditions were examined. Culture-independent methods based on 16S rRNA from fifteen samples revealed that Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes and Acidobacteria were the most abundant phyla. At genus level, were identified Massilia (taxon with more than 50 % of sequences remaining constant during the 30 h of fermentation), Pantoea (taxon with the highest increase during fermentation), Naxibacter, Enterobacter, Raoultella and Klebsiella, forming the carposphere bacterial microbiota of AF. Attributes related to plant growth promotion (siderophores) and others compounds e.g. poly-3-hydroxybutyrate (PHB) and violacein in response to stress conditions could improve the Massilia preponderance. Beta diversity showed that quality parameters of AF (pH, soluble solids, titratable acididy and lipids) and elemental analysis (C, N, H and C/N ratio) were unable to establish microbial patterns in AF. This research offers new insights and perspectives of the indigenous bacterial community composition on AF as a function of spontaneous fermentation during postharvest.Item Acesso aberto (Open Access) Replicação viral, padrão de expressão de citocinas e cinética de morte celular em cultura primária de células neurais infectadas pelo vírus rocio(Universidade Federal do Pará, 2015-07-13) SOUTO, Adriano da Paixão; FRANCO, Edna Cristina Santos; http://lattes.cnpq.br/5939607544965550Arboviruses belonging to the genus Flavivirus (Flaviviridae family) are responsible for considerable morbidity and mortality, may cause severe cases of encephalitis, hemorrhagic fever, hepatitis and febrile disease in vertebrates, including humans. Rocio flavivirus (ROCV) is very important for public health in Brazil, as it represents a serious threat of occurrence of sudden encephalitis outbreaks. The immunopathology of encephalitis caused by flavivirus is not yet fully understood and many aspects involved in modulation of immune responses in the CNS need to be unraveled. Thus, the aim of this study was to analyze the pattern of gene expression of cytokines in primary culture of neural cells when exposed to infection with Rocio. Mixed primary cultures (neuron/glia), obtained from the brains of BALB/C lineage isogenic newborns mice were inoculated with ROCV (MOI equal to 2) on the seventh day of culture and confirmation of infection was made by immunocytochemistry (anti-VROC). Viral replication was quantified in infected primary cultures, depending on the infectious period, by viral titer method. Total cellular RNA was extracted and used for detection of cytokines by real time RT-qPCR techniques. The study showed that primary cultures of neural cells is a good experimental model for the study of CNS infections by Rocio flavivirus. ROCV efficiently infected the primary cultured neural cells leading to cytopathic changes from the 2nd day a.i., when it detected the higher viral titer. The ROCV-infected primary culture survived up to 7 days a.i. and quantification of viral load showed a viral replication kinetics compatible with cell death kinetics of culture infected by ROCV. During the first five days a.i. of the primary culture, there was reduction of IFN-α gene expression, IFN-β, IFN-γ and IL-1β, there was no change in TNF-α expression and there was an increase in TGF-β expression in the 1st, 3rd and 5th day a.i.. The findings of this study suggest that ROCV has the ability to downregulate modulators cytokines of antiviral and inflammatory immune responses and, in this experimental model, immunoregulation exerted by TGF-β predominates over the modulation of inflammatory and antiviral immune responses, larger studies are needed to elucidate immunopathology of CNS infection by ROCV.Item Acesso aberto (Open Access) Produção e avaliação da atividade antioxidante de metabólitos secundários de Piper divaricatum G. Meyer sob diferentes condições de cultivo(Universidade Federal do Pará, 2015-08-06) CORPES, Rosana Silva; SILVA, Joyce Kelly do Rosário da; http://lattes.cnpq.br/2278686174214080Many species of the genus Piper are widely distributed in the Amazon and various biological applications because of large structural diversity of its secondary metabolites. The species Piper divaricatum, is endemic in the Amazon and produces in its essential oil high concentrations of methyleugenol (50-90%), an phenylpropanoid with antioxidant and fungicidal properties. Because of its potential applications, the objective of this study was to establish the in vitro cultivation and comparing the biosynthesis of secondary metabolites and antioxidant properties with the in vivo cultivation. For establish the in vitro culture were used shoot apexes on Murashige e Skoog medium with addition of regulator BAP 0.5 mg.mL-1. For in vivo cultivation, micropiles were propagated in the greenhouse in vermiculite and adding nutritious Murashige e Skoog solution. The volatile compounds identified in the leaves of seedlings grown in vivo were methyleugenol, β-elemene and E-β-ocimene, which did not differ from in vivo cultivation, with the exception of 90 days. The in vitro culture of roots was not efficient to produce phenylpropanoids and presented a very different profile compared to the in vivo cultivation of terpenes. In general, for plants in vitro cultivated there was no statistically significant difference in the phenolics compounds content and antioxidant activity in the leaves. However, the antioxidant activity of roots was significant. The results support the hypothesis that in vitro regenerated plants can synthesize metabolites similar the matrix plant and maintain their biological properties.Item Acesso aberto (Open Access) Ocorrência de inibidores de glicosidases em cianobactérias e microalgas dos estados do Amapá e Tocantins(Universidade Federal do Pará, 2015-08-13) SILVA, Vivian Cassia Oliveira da; XAVIER, Luciana Pereira; http://lattes.cnpq.br/6207860145204571Cyanobacteria are presented as a new source of secondary metabolites for the production of bio-products in many industrial sectors and are used primarily because of its easy cultivation, using few nutritional sources, as well as a large biomass production. Until now we don't have many report in literature of cyanobacteria from amazon, as well as their metabolites and their use in bioproducts. Glycosidases are enzymes of extreme importance as targets for developing drugs that are used for the treatment of various diseases, as well as their inhibitors. This study aimed to identify 24 samples of cyanobacteria and microalgae from the states of Tocantins and Amapá, isolates that produce these inhibitors, as well as extract, fractionate and identify them. Initially, the detection test in petri dishes with esculin the substrate for glycosidase inhibitors has been standardized and the extraction methodology. As a result there was glucosidase inhibitory activity of crude extract and the methanol fraction of four samples (Synechococcus sp, Monoraphydium, P29 and Limnotrix sp), inhibitory activity was detected in the aqueous fractions of any culture. Two methanolic fractions that showed activity was selected for potential assay conducting inhibiting α-glycosidase face with the substrate p-nitrophenyl α-D-glucopyranoside and β-glucosidase to the substrate p-nitrophenyl β-D-glucopyranoside, where both samples had a higher percentage of inhibition for α-glucosidase compared to β-glucosidase.Item Acesso aberto (Open Access) Potencial de produção de antimicrobianos a partir de bactérias isoladas do açaí (Euterpe oleracea)(Universidade Federal do Pará, 2015-08-17) SALLES, Marina Ludmila Oliveira Conor; SANTOS, Agenor Valadares; http://lattes.cnpq.br/9530734927662735Açaí is the fruit of a palm tree widely distributed in the Amazon region and has a high microbial load that favors the fruit fermentation process. It is unknown specific studies to investigate the production of antimicrobial substances of biotechnological interest from microorganisms present in acai fruit, which can result in increased value to the fruit. Since food safety has become an increasingly international concern the application of bacteriocins that target food pathogens without adverse toxic effects, have received great attention. The objective of this work is to isolate bacteria from acai, identify the antimicrobial activity of the isolated bacteria, and characterize the extracts of these bacteria as pH, temperature, stability and action spectrum. The selected fruits were collected in Combu Island and Abaetetuba, State of Para, Brazil. These followed for asepsis and pulp removal and dilutions were inoculated in MRS agar and isolated for analysis The antagonism in vitro was performed with the technique of spot-on-the lawn revealing against three bacteria: Corynebacterium sp, Listeria monocytogenes and Bacillus cereus; and with the well diffusion technique. These extracts were characterized for thermal stability (121 ° C / 15 minutes), enzyme sensitivity to trypsin and pH neutralization. Of the thirty-nine bacterial isolates, it was observed antagonistic activity in nineteen of them. Of thesecould be observed twelve showed extracellular antagonist activity. Four extracts showed sensitivity to trypsin, indicating that these may be bacteriocins. Neutralized extracts showed no activity, which indicates better activity at pH acid extracts. Most of the extracts showed activity after being autoclaved, which may be an indication that the extracts have bacteriocins in Classes I and II. This work shows that bacteria isolated from acai have potential for the production of antimicrobial substances.Item Acesso aberto (Open Access) Estudo do crescimento bacteriano na presença de óleos essenciais de Dysphania ambrosioides l. e Ocimum campechianum mill. para avaliar seus potenciais como antissépticos bucais(Universidade Federal do Pará, 2015-08-31) MOURA, Luiziana Barbosa; SANTOS, Alberdan Silva; http://lattes.cnpq.br/5976702134131016The aromatic plants like Dysphania ambrosioides (mastruz) and Ocimum campechianum (Alfavaca), that are part of folk medicine in Brazil, have phenylpropanoids and terpenoids compounds in their essential oils as result of secondary metabolism that influence the adaptation and defense of these species in environment; particularly, defense against microrganisms. The objective of this study was to evaluate the antimicrobial activity of essential oils of D. ambrosioides and O. campechianum against pathogenic bacteria mouth. For this they used the disk diffusion method on agar and broth microdilution adapted. The plants were obtained in Santa Izabel do Pará, their leaves were washed and weighed; the essential oil was obtained by hydrodistillation. After, the components of essential oils were identified by gas chromatography-mass spectrometry. Bacteria strains used were: Lactobacillus casei (ATCC 7469), Lactobacillus fermentum (ATCC 9338), Streptococcus mutans (ATCC 25175), Streptococcus oralis (ATCC 10557) and Agregatibacter actinomycetencomitans (ATCC 29522). Bacterial suspensions were prepared for testing and 0.5 McFarland standard. As a positive control we adopted the digluconate of chlorhexidine 0.12%. Inhibition halos were observed for all samples in different concentrations of each essential oil (1%, 5%, 10%, 25%, 50% and 75%). The largest halos were found to A. actinomycetencomitans. In the microdilution test the basil oil inhibited the bacteria S. mutans, the main etiological factor for caries in concentration of 1%; mastruz the oil inhibited L. casei, microorganism that enhances the process of decay in concentrations from 10%; Both oils inhibited the growth of A. actinomycetemcomitans, and can be effective against periodontal disease caused by that pathogen. Plant species of this study produce secondary metabolites classes with potential applications in the development of dental products.Item Acesso aberto (Open Access) Relação entre níveis de células T CD4+ e pressão seletiva nos genes env e vif do HIV-1 subtipos B e C(Universidade Federal do Pará, 2015-08-31) PEREIRA, Raimundo Cristovão Ferreira; LEAL, Élcio de Souza; http://lattes.cnpq.br/1158983666415285Previous studies have shown a direct relationship between levels of CD4+ Tlymphocytes and evolutionary rates of HIV-1 (DIAZ et al, 2008; LEMEY et al, 2007; NOSTROM et al, 2014.). Other factors also affect the variability of the env gene: for example function of neutralizing antibodies - Nabs (FROST et al., 2005), the variation in glycosylation sites; (LEAL et al, 2012 LEAL et al., 2008), binding to target cell receptors (i.e, CD4, CXCR4, CCR5), and also the switch from CCR5 to CXCR4 receptor (MILD et al., 2013). Thus, the relationship between viral levels diversification and CD4 + T cells in the env gene can be circumstantial. The vif gene, on the other hand, it is retained and not subject to bias present in the env gene (i.e, glycosylation sites, etc.). Thus, to study the influence of CD4 + T cells levels in HIV variability, the estimate of the selective regimes was used (Nonsynonymous Substitutions - dN and Synonymous Substitutions - dS) by a codon-based model and phylogenetic analysis of unrelated individuals (inter-host). HIV-1 sequences were used of the not-correlated individuals, obtained from the Los Alamos Database. The sequences were separated into CD4+ levels of categories and then analyzed. The analysis revealed no direct correlation between CD4+ T cell levels and evolutionary rates in gp120 and vif gene from HIV-1, subtypes B and C in a population approach.Item Acesso aberto (Open Access) Método de identificação de genes taxonomicamente restritos em dados de RNA-seq em organismo não modelo(Universidade Federal do Pará, 2015-09-28) OLIVEIRA, Lorena Silva de; DARNET, Sylvain Henri; http://lattes.cnpq.br/4586614214029929Black pepper (Piper nigrum) is a biotechnologically interesting plant, research target for both metabolism exploration and improvement related to phytopathological problems, in addition to understanding the evolution of basal angiosperms, ancestral group to which it belongs. With the technological revolution, the next generation sequencing offered access to genetic heritage of non model plants enabling the opening of new biotechnological perspective. The identification of non homologous genes restricted to certain species, called taxonomically restricted genes (TRGs), is a primary biotechnological target, especially in species and groups that are divergent and ancestral. This study aims to establish a method for TRGs identification from RNA-seq data and to validate the approach a dataset for black pepper. The method consists in filtering the transcripts in several stages, so that the annotated transcripts and false positives are removed, and the remaining data without molecular information are classified as potential TRGs. The application of this approach to a black pepper transcriptome dataset (35,631 transcripts) resulted in 22,661 transcripts annotated by similarity. The transcripts that were not annotated in this first analysis were processed in the TRAPID tool, resulting in 12,895 transcripts not annotated. The evaluation of transcripts for false positive detection resulted in 245 true transcripts that were analyzed for the presence of non-coding RNA, resulting in 204 unidentified transcripts. At the end of the method application 71 non annotated transcripts remained with coding regions of protein, indicating potential TRGs. The characterization of these potential TRGs in black pepper can provide new information about the molecular mechanism of this specie and perhaps elucidate pathways for the establishment of cultivars tolerant to disease.Item Acesso aberto (Open Access) Preparação de suportes micro e mesoporosos para imobilização de lipase e aplicação na esterificação do ácido oléico(Universidade Federal do Pará, 2015-09-29) QUARESMA, Francisco Lucio Barbosa; NASCIMENTO, Luis Adriano Santos do; http://lattes.cnpq.br/3720461233595226This study aimed to prepare and characterize heterogeneous catalysts using immobilized enzymes in MCM-41 supports and metakaolin. The supports were functionalized with APTES (an amine) for application to the oleic acid esterification reaction with methanol to produce ester. The characterization of the catalyst was made by XRD, FT-IR, physisorption of N2 and spectrometry of UV. The results show that MCM-41 has been effectively functionalized and immobilized enzymes were, however, there was a partial loss of the hexagonal array of such support, while for metakaolin, both functionalization as immobilization did not occur satisfactorily. Using UV spectrometry, it was found that around 80% of the enzymes were effectively immobilized on MCM-41. This biocatalyst (heterogeneous) was tested in the esterification of oleic acid (molar ratio to oleic acid / methanol was 1:15), at a temperature of 30 °C (100 rpm) for 12h. The obtained conversion was 52% of acid mass to ester, indicating that there viability on the use of MCM-41 amine functionalized for the immobilization of lipase, since catalytic activity of the enzyme was preserved. As for metakaolin, the results showed that, despite having been initially some connection enzymes to metakaolin functionalized, UV spectrometry showed that the anchoring enzyme was not effective in the course of 4 hours, however, new methodological approaches to this material may be applied to ensure the efficient functionalization and immobilization.