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Ação da ciclosporina A na via de ativação do fator de crescimento de nervo (NGF) em células neurais do SNP
(Universidade Federal do Pará, 2016-08-26) JESUS, Jessica Batista de; SENA, Chubert Bernardo Castro de; http://lattes.cnpq.br/8620752020290438; NASCIMENTO, José Luiz Martins do; http://lattes.cnpq.br/7216249286784978
A is an immunosuppressive drug with known action on T cells of the immune system used in organ transplantation and autoimmune diseases. In the nervous system, cyclosporin A acts by inhibiting the action of Calcineurin, an important second messenger from pathway of signal transduction Nerve Growth Factor (NGF), resulting in hyperphosphorylation of the nuclear factor of activated T cells (NFAT), and downregulation of NGF, TrkA and other factors that participating in this pathway. The NFAT1-4 family are dependent isoforms of calcineurin, while NFAT5 isoform is independent. It has been demonstrated the neuroprotective role of Cyclosporin A via calcineurin dependent or independent. In this study, we evaluate the action of Cyclosporine A in the PNS system, that could be associated with levels of NGF, TrkA and an independent of calcineurin transcription factor (NFAT5) that interplay the plasticity of neuronal cells derived from Dorsal root ganglia (DRG) maintained in cultures. We use E10 DRG cultures supplemented with medium conditioned E9 Retinal treated with Cyclosporin A for 48 and 72 hours. Cultures enriched neurons were confirmed by calcium imaging method. The action of Cyclosporine A in the neuritogenesis was assessed by bright field microscopy, expression of NGF, TrkA and NFAT5 was performed by RT-PCR, intracellular accumulation of NGF was evaluated by immunofluorescence and the presence of TrkA in neurons. The viability test of the cultures treated or not with the concentrations of 1-40μM Cyclosporine A was performed by MTT method. The results show an increase of NGF levels in mixed cultures, and TrkA receptor and NFAT5 in cultures enriched in neurons following treatment with cyclosporine A. Given the importance of NGF pathway in the development and maintenance of the SNP, the use of Cyclosporin A have activity in the peripheral nervous system cells, which might be used in the clinic with new target for new therapies.
Acesso aberto (Open Access)
Análise citogenética de duas espécies do gênero Hylaeamys (Rodentia: Cricetidae) por citogenética clássica e molecular
(Universidade Federal do Pará, 2013-04-05) PINTO, Jamilly Amaral; NAGAMACHI, Cleusa Yoshiko; http://lattes.cnpq.br/8887641213110093
Rodents are one of the largest and oldest orders of the class Mammalia. In South America, the order Rodentia compromises about 42% of mammal species, and from this more than 50% belong to the family Cricetidae, which includes the subfamily Sigmodontinae. The genus Hylaeamys is inserted in the tribe Oryzomyini and corresponds to one of 10 new genera proposed for species and species groups within Oryzomys. Hylaeamys is the equivalent of "megacephalus group", and consists of the species H. acritus, H. laticeps, H. megacephalus, H. perenensis, H. oniscus, H. tatei and H. yunganus, distributed in Venezuela, Trinidad, Guyana, Paraguay and Brazil, in areas of the Amazon rain forest, Atlantic rainforest and savannah. This study aims to analyze chromosomal markers in two species of the genus Hylaeamys, providing data to assist in its taxonomic and cytogenetic characterization. Nineteen samples of Hylaeamys megacephalus (HME) and four samples of Hylaeamys oniscus (HON) were analyzed. HME has 2n = 54 and HON, 2n = 52. The results obtained by G- and C-banding and Fluorescent In Situ Hybridization with whole chromosome probes from Hylaeamys megacephalus made it possible to determine the chromosomal characteristics of the species studied, as well as allowing a comparative analysis between them, and in comparison with Cerradomys langguthi, observing homeologies and karyotypic differences. The two species of Hylaeamys differ by a centric fission/fusion rearrangement in which HON shows the association of the pairs 14/19 of HME. This association is shared with CLA with an inversion (19/14/19). This work is an achievement for phylogeny and chromosomal studies on the genus Hylaeamys.
Acesso aberto (Open Access)
Análise conformacional da enzima protease do HIV-1 relacionada à resistência ao inibidor Nelfinavir
(Universidade Federal do Pará, 2017) HOLANDA, Luiz Henrique Campos; SILVA, Jerônimo Lameira; http://lattes.cnpq.br/7711489635465954; SOUSA, Maisa Silva de; http://lattes.cnpq.br/1775363180781218
The Human Immunodeficiency Virus (HIV), which causes acquired immunodeficiency syndrome (AIDS), is a retrovirus that has highly virulent glycoproteins that invade the CD4 + T lymphocyte through its CCR4 and CXCR5 receptors. The biological cycle of HIV is mediated by the protease, transcriptase and integrase enzymes. HIV-1 protease is an enzyme that is present in the final phase of the biological cycle, where virus maturation occurs, and is an important pharmacological target. The main objective of this project is to verify the effects of the D30N, I84A and M46I mutations on the HIV-1 protease enzyme and the complex formation with the nelfinavir inhibitor through molecular dynamics and bioinformatics techniques. The results based on the structural analyzes showed structural differences between the studied systems. The 1OHR system presented a closed conformation, the systems D30N and D30N_I84A_M46I presented semi-open conformation and the D30N_I84A system presented open conformation, in which the latter presented lower free energy value and greater instability in the RMSD analyzes, however the greater flotation of residues Of amino acids. The theoretical analyzes showed the importance in the resistance of the double mutation D30N_I84A and the conformational restructuring capacity of the M46I mutation and catalytic capacity.
Acesso aberto (Open Access)
Análise da atividade enzimática de quitotriosidase como um marcador para a malária vivax: abordagens bioquímicas e moleculares
(Universidade Federal do Pará, 2010) CRUZ, Cleber Monteiro; SILVA, Luiz Carlos Santana da; http://lattes.cnpq.br/6161491684526382
Chitotriosidase was the first described chitinase and its physiologic role is not entirely clear, although many studies have been showed its participation as a component of human immune response. A 24pb duplication on exon 10 of chit1 gene results on RNAm frameshift, leading to a 87 nucleotides deletion. This alteration generates a protein with no catalytic activity at all. This condition is called chitotriosidase deficiency and presents a frequency close to 6% of homozygosis duplication in different ethnical groups. Malaria is an amazon endemic parasitosis caused by protozoaries of genus Plasmodium and causes symptoms as fever, headache and vomit, which leads to a characteristic immune response. The objective of this study was to evaluate the chitotriosidase enzyme behavior in patients suffering of malaria in Pará state and to determine the frequency of 24pb duplication on chitotriosidase gene in a representative sample. Chitotriosidase measurement was made in 100 healthy individual and in 47 malarial patients. The molecular analysis of the 24pb duplication was realized in 100 volunteers trough a protocol which included DNA extraction techniques, PCR and 2,5% agarose gel visualization to verify normal fragments (normal homozygote: 195pb) and the 24pb duplication (mutant homozygote: 219pb; heterozygote: 219pb e 195pb). This study described at first time on scientific literature the chitotriosidase plasmatic levels increasing in patients suffering of malaria vivax compared to healthy individual. No association was observed between parasitemia and plasmatic chitotriosidase levels in malarial patients. Molecular analysis showed a frequency of 72% normal homozygotes, 24% heterozygotes and 4% mutant homozygotes to 24pb duplication. Allelic frequencies were around 84% to wild allele and 16% to mutant allele. No correlation was found between genotype and biochemical phenotype (represented by chitotriosidase levels) on control group.
Acesso aberto (Open Access)
Análises moleculares da região controle do DNA mitocondrial de astrocitomas na população paraense
(Universidade Federal do Pará, 2012-06-06) COSTA JÚNIOR, Carlos Antonio da; ANSELMO, Nilson Praia; http://lattes.cnpq.br/6518287721873199
The central nervous system cancer represents 2% of all malignancies in the world population and 23% of cases of childhood cancer. In Brazil, an estimated 4,820 cases of cancer in men and women in 4450 to the year 2012. Gliomas are tumors of the central nervous system formed from glial cells, making up over 70% of brain tumors. The most important property of gliomas is the ability of immune evasion. Age, ethnicity, gender and occupation may be considered risk factors for the development of gliomas, and are twice as common in African-Americans. The astrocytoma is the most common glial tumor, constituting about 75% of cases of gliomas. These tumors are classified into four levels according to the World Health Organization. Mitochondrial DNA is related to the development and progression of various types of tumors. Mitochondrion is responsible for cellular energy balance and is involved in triggering apoptosis responding to oxidative stress. Mutations in DLOOP can change DNA replication rates and increase the developing cancer risk. We analyzed 29 samples astrocytoma classified according to the WHO. Our data suggest that low-grade astrocytomas may be related to genetic inheritance, making some patients with specific mutations or polymorphisms more susceptible to the risk of developing the disease, and high grade may be related to prolonged exposure to carcinogenics. Polymorphisms and mutations have been identified which correlate with some risk of developing astrocytomas and disease progression. The insertion of two or more nucleotides at microsatellite regions may cause instability and contribute to the cancer onset. Deletion at the site 16132 may be a high-grade astrocytoma marker, as well as insertion of two or more cytosines to the site 16190 can be an astrocytoma specific marker. Heteroplasmy may be decisive for the emergence and / or progression of high-grade astrocytomas.
Acesso aberto (Open Access)
Avaliação de biomarcadores sorológicos em um estudo de busca ativa de casos novos de hanseníase em área hiperendêmica
(Universidade Federal do Pará, 2016-10-07) GOBBO, Angélica Rita; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
Leprosy is a cronic infection diasease clinically characterized by changes in tactile, thermal and painful sensitivity in skin and peripheral nerves. Due to the absence of laboratory diagnosis of leprosy, new tools that contribute for identification of cases are necessary for enable patient treatment before progression to physical disabilities. In this sense, the present study aimed evaluate serological biomarkers contribution for early diagnosis of leprosy. Was perfomed an active case finding study in Mosqueiro district, Belém – Pará. All individuals were clinically examined by experient leprologists doctors and than 5mL of peripheral blood were colleted for future titration of anti-ND-O-BSA, anti-LID-1 e anti-NDO-LID by ELISA. The action of active finding in Mosqueiro district diagnosed 104 new cases of leprosy between 895 subjects examined (11.6%), indicating a high hidden endemy that agree with the high seroprevalence between schoolchildren. Were observed a significant difference among patients with late or early diagnoses, mainly in multibacillary forms. All biomarkers tested showed promising results in detection of late cases, such as related in literature, however, for early cases those molecules identified correctly only 50% of patients. None of biomarker tested presented sufficient sensitivity to detect all leprosy patients, early or lately diagnosed. Besides, LID-1 molecule had evidenced a lower sensitivity for early cases, their high especificity and accuracy suggest their use as a potential tool for serological screening to identify assintomatic subjects with high risk of illness. Thus, we concludes that besides no biomarker had reveled utility as a serological diagnostic tool, the detection of anti-LID-1 presented a possible aplicability as a screening marker of subjects with increased risk to develop leprosy, contributing indirectly for leprosy diagnosis.
Acesso aberto (Open Access)
Caracterização do padrão de expressão e metilação do gene P21CDKN1A/CIP1 em tumores mamários caninos
(Universidade Federal do Pará, 2018-04-20) SOUSA, Raissa Melo de; BORGES, Bárbara do Nascimento; http://lattes.cnpq.br/0676220027193876
The most of canine mammary tumors are malignant and associated with the animal death. One of the factors involved in this pathogenesis is the change in the expression level of the gene P21, which in turn encodes a protein that can inhibit tumor initiation and tumor progression. The methylation profile of gene can affect the cellular level of p21 and lead gene silencing or overexpression. Whereas the functional importance of the P21 gene, this study aimed to evalu-ate the methylation profile and expression in mammary tumors of dogs, in order to identify molecular markers of early diagnosis, survival and prognosis. Therefore, 83 tumor and non-tumor tissue samples were collected from dogs, undergoing surgery at the Veterinary Hospital "Mário Dias Teixeira”, in Belém -Pará. the DNA and RNA from each sample were submitted to extraction using a commercial kit. For the methylation analyzes, the obtained DNA was sub-mitted to the modification process, with a subsequent technique of Bisulfite Sequencing PCR, using region-specific primers and subsequent visualization in 2% agarose gel. The sequencing results were analyzed in BiQ Analyzer software in order to evaluate the methylation pattern. For gene expression analysis, the target gene mRNA was quantified using the real-time PCR technique using the GAPDH and HPRT1 genes as constitutive controls. Statistical data was performed using the Fisher Exact, Odds Ratio and Mann-Whitney tests in the GraphPad Prism program, considering the significant results when p ≤0.05, with a 95% confidence inter-val. In order to evaluate the methylation profile, the CpG islands of the P21 gene were characterized. The island 1 is located in an intron with 34 CGs, while an island 2 was identified in the exon, with 22 CGs. Both amplified generating a fragment of ~ 300bp. At island 1 no methylation was detected, whereas island 2 was methylated, but the island methylation profile was not different between the tumor, non-tumor and control samples. With these results was impossible compare the methylation values with clinical and expression data, suggesting that these regions are not altered. The expression levels of the tumor samples were low when compared to the non-tumor samples and control, with p (0.0001), show that the role of p21 in these tumors may be altered but not statistically significant when correlated with clinical histopathological data of patients. However, a reduced expression in the survival of animals> 1 year of age was observed, and a high expression in animals with survival <1 year of age, suggesting the influence of p21 as a marker of prognosis. More detailed studies of the P21 gene are still needed to see if these changes could be used as molecular markers, and thus aid on the prognosis and detection of cancer in the species.
Acesso aberto (Open Access)
Detecção de vírus Influenza A em aves migratórias capturadas em regiões litorâneas dos estados da Bahia, Pará e Pernambuco
(Universidade Federal do Pará, 2014) FERREIRA, Deimy Lima; MELLO, Wyller Alencar de; http://lattes.cnpq.br/1784167608719139; SOUSA, Rita Catarina Medeiros; http://lattes.cnpq.br/3560941703812539
The Influenza virus is known for its ability to infect a wide variety of animals, such as mammals (humans, pigs, horses, whales), domestic birds (chicken [Gallus gallus], goose, turkey [Meleagris ocellata]) beyond wild birds of the orders Anseriformes (duck, wild goose and swan) and Charadriiformes (seagulls, swallows, aquatic birds and sandpipers) these being its natural host. Comprehend the movement of long distance migratory wild birds is crucial to explain the movement of avian influenza viruses. This event causes movement of the birds are acting as an important means of spreading the virus along a migratory route, a fact widely accepted. During the period 2006 and 2007, samples were collected 2,252 samples from a variety of bird species captured in locations which are part of the Atlantic migration route and Mississippi, in the states of Bahia, Para and Pernambuco for epidemiological surveillance of West Nile virus and influenza. The objective of this study was to investigate the circulation of influenza virus among migratory birds that use the routes that pass the above states. For this, the samples were analyzed by means of molecular biological techniques, which comprised two main steps: a) extraction of DNA / RNA from the biological specimen; b) amplification of the gene encoding cytochrome oxidase control of by the technical Polymerase Chain Reaction (PCR) and amplification of the vRNA by Real time Reverse Transcripition polimerase chain reaction (RT-qPCR). The results obtained showed that the total samples tested, 7.2% (n = 158) were positive by RT-qPCR for Influenza A virus. We observed a difference in positivity for the virus among bird species analyzed, which is 3.58% for Charadriformes order, 26.3% among the birds of the order Anseriformes, 5.3% of birds belonging to the order Pelecaniformes and 10.9% for those order Suliformes. Among the samples of the orders Passeriformes and Columbiformes, no sample was positive for Influenza Virus. The data suggest variation among the sampling sites, and the state of Para with the lowest percentage of positivity, the second highest rate with Bahia and Pernambuco finally presenting higher prevalence of absolute value. This study shows that although rare investigations in Brazilian territory, there has been movement of Influenza A viruses among several species of migratory birds that utilize the states of Para, Bahia and Pernambuco as stopping places and reproduction of their species. These findings justify further investigations to understand the dynamics of avian influenza viruses circulating in the population of wild birds in Brazil, and its role as a potential source of infection for other animals, including humans.
Acesso aberto (Open Access)
Diagnóstico citológico e molecular da infecção pelo HPV em mulheres do município de Barcarena, Pará, Norte do Brasil
(Universidade Federal do Pará, 2010-06-02) NASCIMENTO, Ludmila Marcia Sousa do; BRITO JUNIOR, Lacy Cardoso de; http://lattes.cnpq.br/9705670940390281; SALGADO, Claudio Guedes; http://lattes.cnpq.br/2310734509396125
HPV (human papillomavirus) was appointed by the WHO (World Health Organization - WHO) as the main risk factor for developing cervical cancer, thus becoming an important and very serious public health problem, especially in underdeveloped countries or under development. The early sexual activity, multiple partners and casual sex, smoking, immunosuppression (eg, the population of AIDS patients), pregnancy, sexually transmitted diseases like herpes and chlamydia prior, in addition to non-compliance with the measures already taken to prevent Sexually Transmitted Diseases (STDs), such as the simple use of condoms is admittedly associated with the incidence of HPV infection. This research aimed to evaluate the diagnostic performance of the methodologies of conventional cytology (Pap test) compared to liquid based cytology, and to determine the prevalence of genotypes 16 and 18 of HPV in women without HPV cytopathic effect compatible with and relate to presence of inflammatory conditions, or not associated with HPV, with epidemiological data such as age, education, socio-cultural condition of women from the municipality of Barcarena - Pará - Brazil. To do so, in this study voluntarily, 50 women attended at the Health Unit Barcarena - Brazil, through the campaign to collect Pap test as a method of preventing cancer of the cervix. These women were informed about all procedures performed by the body of this health study and the results of this search and only after the volunteers have signed the Deed of Consent, they were included for sampling. The analysis and test results of liquid based cytology and conventional were performed according to the Bethesda Classification and reviewed blindly by two pathologists. For the analysis statistic was used Fisher's exact test and the "Screening Test" to determine the specificity / sensitivity of the methods, considering the significant value of p ≤ 0.05. Our results indicate that the use of liquid based cytology has demonstrated a number of advantages over conventional cytology. In molecular diagnostics (PCR) were observed occurrences of HPV types 16 and 18 in 10% of women attended. Among the cases that were PCR positive for types 16 and 16/18 most of the women were 27.4 years old on average, with more schooling, performing household chores and rural areas, and with instances of co-infection by infectious agents cause other sexually transmitted diseases. The results of this study reinforce the importance of maintaining free campaigns to prevent cervical cancer as a preventive measure in combating this disease, especially in Pará State, where, probably, the epidemiological profile of disease is associated with the large distances that women in river communities have to travel to perform this test free of charge, the type of economic activity in the region, the local bias still exists with the test, and the degree of difficulty of implementing effective return of patients to medical appointments after obtaining the test result and the same routing for molecular diagnosis of positive cases for type lesions ASC-H and CIN I, II and III.
Acesso aberto (Open Access)
Estudo da influência do uso de agrotóxicos e de polimorfismo do gene GSTT1 na etiologia de fissuras labiopalatais em pacientes do Estado do Pará
(Universidade Federal do Pará, 2014-08-28) MARTINS, Cláudia Maria da Rocha; BURBANO, Rommel Mario Rodriguéz; http://lattes.cnpq.br/4362051219348099
Birth defect or congenital malformation is any anatomic, metabolic or functional abnormality, inherited by a mechanism of Mendelian transmission or caused by a new mutation, a chromosomal alteration or physical aggression by an infectious, chemical or the fetus or embryo development . Its causes may be genetic or environmental, and, most often, are multifactorial origin, through the genetic predisposition factors interact with environmental factors triggers. In Pará state, a large number of individuals affected by Oral clefts are from rural areas, mainly in the northeastern state where it is notoriously indiscriminate use of pesticides harmful to human health, many of which have high potential teratogenic. The objective of our study was to investigate the association between the polymorphism (rs4630) in the GSTT1 gene and exposure to pesticides in etiology of oral clefts and analyze the pattern of changes in speech of patients according to the type of cleft. Eight three patients with cleft palate or lip, and of both sexes, and 83 mothers of these patients, all from the state of Pará, residing in rural and capital area were analyzed. Speech therapy and analyzes were performed with the blood of these individuals, the molecular analysis was performed. Statistical analysis was performed using the statistical software SPSS v.. 12.0 and BioEstat v. 5.0. Tests included testing multiple logistic regression test x2e the Fisher exact test. Our result consists of five different molecular analyzes. We found that the presence of the C allele in the genotype of the individual can influence the metabolism of xenobiotics and increase the risk to develop oral clift.
Acesso aberto (Open Access)
Estudo do perfil de pacientes submetidos a pesquisa de Helicobacter pylori: análise endoscópica e dos fatores determinantes da atividade linfocitária na resposta imunológica gástrica (ROR-Y, FOXP3 e GATA3)
(Universidade Federal do Pará, 2015) MIRANDA, Ariney Costa de; QUARESMA, Juarez Antônio Simões; http://lattes.cnpq.br/3350166863853054
INTRODUCTION: Helicobacter pylori is known for its adaptability to the host may progress to chronic infection using diverse and effective mechanisms of pathogenicity. It has high worldwide incidence and its direct relationship with peptic ulcer, gastritis, gastric carcinoma and lymphoma occurs in a minority of infected individuals. A better understanding of the genetic regulation of gastric immune response, motivated this investigation. OBJECTIVES: Describe the transcription factors of T lymphocytes positive for ROR-γ, FOXP3 and GATA3, correlating them with the intensity, type and degree of activity of gastritis, caused by H. pylori infection METHODS: The study included 50 patients of both sexes who underwent upper gastrointestinal endoscopy with biopsy. Urease test and histology were made for identification and confirmation of infection by the bacteria. Thirty-five samples were sent to the immunopathology laboratory NMT-UFPA to study gene expression of transcription factors of T lymphocytes (ROR- γ, FOXP3 and GATA3) by RT-PCR method. RESULTS: We obtained 48.5% positive H. pylori urease test in patients and 25.7% positive of H. pylori in the histological study. The confirmation of H. pylori held by these two exams was 11.7%. In this sample, 22% of individuals reported having a previous Helicobacter pylori infection. The age and gender of the individuals did not influence the gene expression of the studied factors. The H. pylori positive individuals showed a higher expression of the GATA3 gene with prior infection report, and mild erosive gastritis body classified by the Sydney system via endoscopy. The ROR-γ gene presented with increased expression only when comparing samples with or without positive for H. pylori (histology), by the topography of the inflammatory process evidenced by endoscopy. The terms of the factors in the study were more significant when we used the β-actin gene as standard when compared to the GAPDH gene. CONCLUSIONS: The adult age group analyzed in our sample did not influence the gene expression of the studied transcription factors. 2- There were not found differences in the genes expressions that were studied, related to gender of the sample. 3- There was a significant gene expression not only in the patients that were H. pylori positive (histology), but also in the ones who reported previous infection in our study. 4-To compare the endoscopic findings of the sample using the Sydney system with the gene expression of transcription factors under study, we obtained better agreement only in the degree of activity of gastritis. 5- The transcription factor GATA3 (TH2 response profile) was the highest gene expression in samples with endoscopic gastritis and tested positive for H. pylori. 6- The transcription factor ROR-γ (TH17 response profile) presented with increased expression when comparing samples with the topography of the inflammatory process evidenced by endoscopy, regardless of positive H. pylori (histology). 7- The gene β-actin gene as a constituent standard used in our study was that showed significant results in quantified terms, when compared to the GAPDH gene.
Acesso aberto (Open Access)
O fator de crescimento neuronal na infecção por Schistosoma mansoni: estudo molecular, imunoenzimático e morfométrico em modelo permissível e não permissível à infecção
(Universidade Federal do Pará, 2013-07-03) SANTOS, Daniel Valle Vasconcelos; SILVA FILHO, Manoel da; http://lattes.cnpq.br/2032152778116209
Schistosomiasis is a tropical disease caused by Schistosoma mansoni. His occurrence affects 110 million people worldwide. The deposition of eggs of the parasite may occur - in ectopic form – in the central nervous system (CNS) which leads to the formation of granulomas with consequent production of nerve growth factor (NGF). Since several studies have demonstrated the importance of NGF in the development of visual cortical pathways, our study aimed at evaluating the possible changes in the NGF concentratons in the visual system as well as the impact of this on the pyramidal cell morphology in two animal models. The change in concentration of the nerve growth factor as well as neuronal morphology were evaluated in suscetible and non-suscetible animals (mice and rats) to infection. We used 174 rats (Hooded Lister) and 135 albino mice bred and kept in cages and fed ad libitum. These animals were infected shortly after birth, with 50 cercariae. Seventy seven rats and 73 mice were inoculated with saline and constituted the control group of the study. The infection covered a period of 48 weeks . Samples of liver and visual cortex were removed, extracted and quantified with immunoassay kit (ChemiKineTM Nerve Growth Factor (NGF) Sandwich ELISA Kit - Chemicon International). For the morphometric analysis we used the pyramidal cells of the visual cortex layer IV marked by extracelular injection of biotinylated dextran (10,000 kDa). The results were expressed as mean ± standard deviation. We used Student t test to determine statistical differences between groups. The average value of NGF found in the visual cortex of rats infected was 39.2% higher than in the control group (infected: 400.9 ± 143.1 pg/ml, control: 288 ± 31.9 pg/mL, p < 0.0001). In liver samples, the increase was 28.9% higher in the infected group (infected: 340.9 ± 103.9 pg/mL, p < 0.01, control: 264.4 ± 38.6 pg/mL). No significant increase was detected within a week of infection. Among the mice group, the increase of NGF in the visual area was 94.1% (infected: 478.4 ± 284 pg/ml, p < 0.01; control: 246.5 ± 76.8 pg/ml). In the liver of these animals the increase was 138.7% (infected: 561.8 ± 260.7 pg/mL, p < 0.01, control: 301.3 ± 134.6 pg/mL). In mice group we found significant differences in dendritic parameters evaluated. The number of dendrites was 11.41% higher in the infected group than in the control (control: 25.28 ± 5.19; infected: 28.16 ± 7.45, p < 0.05). The total length of dendrites was also affected (control: 4916.52 ± 1492.65 μm; Infected: 5460.40 ± 1214.07 μm; p < 0.05), representing an increase of 11.06%. The total area of the dendritic receptive field was increased by 12.99% (control: 29.346,69 ± 11.298,62 μm2; Infected: 33.158,20 ± 7.758,31 μm2, p < 0.05) while the area had a somatic reduction of 13.61% (control: 119.38 ± 19.68 μm2; infected: 103.13 ± 24.69 μm2, p < 0.001). When we evaluated the effects of increased NGF in rats infected we did not observe significant differences in dendritic parameters analyzed, compared to the control group, except for an increase in the area of the neuronal body of approximately 21.18% (control: 132,20 ± 28.46 μm2; infected: 160.20 ± 31.63 μm2, p < 0.00001). This work showed that the reaction production of NGF in the CNS during infection with Schistosoma mansoni occurs in greater magnitude than permissible in the model in the model impermissible. We also demonstrated that in mice the effects on neuronal morphology is dramatically affected when the body is subjected to an increase in the concentration of NGF as a result of infection by Schistosoma mansoni. Given these data, studies evaluating the potential impact of visual effects and also in cell physiology caused by schistosomiasis infection becomes necessary to assess the actual damage caused by this pathological increase of nerve growth factor in the visual pathways of mammals.
Acesso aberto (Open Access)
Imunoreatividade para os receptores de neurotrofinas P75NTR e TrkA na zona subventricular de ratos adultos após isquemia estriatal
(Universidade Federal do Pará, 2015-08-21) TAVARES, Patrycy Assis Noronha; LIMA, Rafael Rodrigues; http://lattes.cnpq.br/3512648574555468; LEAL, Walace Gomes; http://lattes.cnpq.br/2085871005197072
Neurotrophins are growth factors expressed by cells of the nervous system both during development and in adulthood. The Nerve Growth Factor (NGF, the English- Nerve Growth Factor), brain-derived neurotrophic factor (English- BDNF- of Brain-Derived Neurotrophic Factor), Neurotrophin-3 (NT-3), Neurotrophin-4/5 ( NT-4/5), have many functions related to aging and response of nervous tissue to the pathology such as vascular accident (CVA). In this pathology, the increase of the neurotrophin expression can interfere with the degree of neurogenesis in the sub-ventricular zone (SVZ) and redirect the rostral migratory flow of Adult Neural Stem Cells (CTNAs) to the ischemic region. The presence of neurotrophin receptors TrkA and p75NTR in the CTNAs of SVZ indicates that they may participate in the regulation of neurogenesis in this region. Here we describe the influence of an experimental ischemia by microinjection of a vasconstritor Endothelin-1 peptide, which is restricted to the striatum adjacent SVZ; on the pattern of immunoreactivity for TrkA and p75NTR receptors in different survival times. The histopathological pattern of ischemic striatum and the cytoarchitecture of the SVZ, followed by immunohistochemical analysis to the receptors were analyzed. Numerous p75NTR + cells were found in the ipsilateral SVZ and against the injection site, with had a reduction in immunoreactivity at first and third day after ischemia. Few TrkA + cells were found in SVZ of both groups, however, many TrkA + axonal terminals were saw in the ischemic ipsilateral SVZ. Soon after the ischemic process, there was thickening of the SVZ, the concomitant reduction in immunoreactivity for p75NTR and TrkA + arisings of axonal terminals.
Acesso aberto (Open Access)
Inibição da atividade da Tirosinase por análogos do ácido Kójico
(Universidade Federal do Pará, 2014-11-14) CARDOSO, Erica de Tássia Carvalho; MAUÉS, Luis Antônio Loureiro; http://lattes.cnpq.br/4851018582496177; NASCIMENTO, José Luiz Martins do; http://lattes.cnpq.br/7216249286784978
Tyrosinase is an enzyme’s key for melanin biosynthesis. It is a "copper-dependent" enzyme which exhibits three intermediate states: deoxy (Cu1+ -Cu1+), oxi (Cu 2+ - O2 -Cu2+) e met (Cu2+) - Cu2+). This enzyme has bifunctional activity since it can oxidize phenol or catechols in their corresponding o-diphenols. In addition, oxidation of phenols can be described by Michaelis-Menten kinetics. Hyperpigmentation disorders and enzymatic browning of fruit and fungi is associated with tyrosinase.Therefore the research for substances of natural or synthetic origin that could have an effective regulation on the behavior of this enzyme is a key factor of the treatment of such disorders. In this perspective, the present study consisted in analyze the biochemically anti-tyrosinase activity of kojic acid and its analogues derivatives from 4H-pyrones (S-01, S-02, S-03 and S-04) and derivatives from diidropirano [3, 2-b] cromenodionas (S-05, S-06, S-07 and S-08) chemically designed by molecular modeling in LPDF from ICEN UFPa. The kinetics of substances S-02, S-04, S-06, S-07 and S-08 showed competitive inhibition, similar to the pattern of inhibition of kojic acid with Ki values = 145,0 ± 20,0 μM; 64,0 ± 10,0 μM; 4,0 ± 0,0 μM; 6,0 ± 0,0 μM; 9,0 ± 0,0 μM, respectively, and 5,0 ± 0.0 μM for kojic acid, while S-01 had mixed type of inhibition (Ki = 999,0 ± 150,0 μM). Since the S-03 and S-05 substances showed no inhibitory activity. The substances tested demonstrated a high degree of safety both in the integrity of the erythrocyte membrane in the hemolysis test as in the test with MTT viability in cultures of MRC5 fibroblasts, culture of nerve cells from chicken embryo retina and B16F10 melanoma. Thus, it was demonstrated that S-02, S-04, S-06, S-07 and S-08 substances have potent activity as inhibitors of tyrosinase and may be candidates for the treatment of pigmentation disorders.
Acesso aberto (Open Access)
Investigação da prevalência da amebíase em escolares do município de Imperatriz-MA
(Universidade Federal do Pará, 2012) BELFORT, Marcia Guelma Santos; ISHIKAWA, Edna Aoba Yassui; http://lattes.cnpq.br/3074963539505872
Amoebiasis is an infection caused by Entamoeba histolytica and an important cause of morbidity and mortality worldwide. Studies have reported a high prevalence of in tropic amebiasis al regions, especially in communities living in poor sanitary conditions. The epidemiological study of amoebiasis has been reevaluated since E. histolytica, pathogenic form, was distinct from E. dispar, non-pathogenic form. The aim of this study was to estimate the prevalence of E. histolytica in the population of the municipal city Imperatriz (MA). We conducted a cross-sectional study involving 405 students. By the screening of the complex E. histolytica / E.dispar parasitological examination was performed using the sedimentation method. The positive samples for E.histolytica/ E.dispar complex were subjected to polymerase chain reaction (PCR) for species differentiation. For initial amplification by the PCR we used a outer primer set E1 and E2 that amplified a 1076 bp fragment and followed by a multiplex PCR using inner primer set Ed-L/Ed-R and Eh-L/Eh-R E.histolytica and E.dispar respectively. No sample showing positivity for E. histolytica The prevalence of E.dispar in the population was 2.7% (11/405). The PCR proved important tool for the differential diagnosis of Entamoebas. However, studies on the prevalence of amoebiasis should be conducted in population with different characteristics, in order to contribute effectively to define the epidemiological situation of this infection in Imperatriz city.
Acesso aberto (Open Access)
Isolamento de microssatélites de espécies madeireiras no contexto da sustentabilidade genética no manejo florestal
(Universidade Federal do Pará, 2004-05-10) VINSON, Christina Cleo; Yamaguishi, Ana Yamaguishi; http://lattes.cnpq.br/7012636819010752; SAMPAIO, Maria Iracilda da Cunha; http://lattes.cnpq.br/2482763145819602
Aborda isolamento de microssatélites de espécies madeireiras no contexto da sustentabilidade genética no manejo florestal com o objetivo de isolar e caracterizar uma bateria de marcadores microssatélites para espécies arbóreas da Floresta Amazônica: Jacaranda copaia, Bagassa guianensis e Dipteryx odorata. Estes estudos vem sendo realizados como parte do projeto Dendrogene, e as análises genéticas populacionais posteriores fornecerão subsídios para conservação e manejo sustentável destas espécies.
Acesso aberto (Open Access)
Perfil das mulheres ribeirinhas com infecção pelo Papilomavírus humano, com ênfase nos tipos 16 e 18, em populações da Amazônia oriental
(Universidade Federal do Pará, 2015) DUARTE, Daniel Valim; SOUSA, Maisa Silva de; http://lattes.cnpq.br/1775363180781218
The human papilomavírus (HPV) is the cause of the most common sexually transmitted infection in the world and is recognized as the main causative agent of cervical cancer, but little is known about the prevalence of this virus in isolated communities in the Amazon. We aimed in this study to identify among women in riverside communities of several municipalities in the state of Pará - Brazil, the general and specific prevalence of HPV infection and HPV types 16 and 18, building an epidemiological profile of women positive for the virus and these types. Between February and December 2008, cervical samples riverside women were collected on active serach for the Pap smear testing. In these samples, molecular analysis of HPV were performed by conventional polymerase chain reaction (PCR) followed by real-time PCR to diagnose types 16 and 18. Of the 353 samples tested, 58 (16.4%) were positive for HPV , of these 8 (2,3%) were identified positive for type 16 and 5 (1,4 %) were positive for type 18 with one case of infection by both types investigated . Women under 16 years had 34.6 % prevalence had rate and women between 52 and 66 years rate of 29.8 % (p=0.003), women who reported having only one fixed sexual partner had lower prevalence of viral infection (11,8%) (p<0,001). There was a relative increase in viral presence in accordance with the severity cytological shown (p = 0.026). There was no statistical association between viral types 16 and 18 with the variables investigated. The presence of HPV and its most associated with cervical cancer types highlights the importance of specific actions aimed at the prevention, transmission and screening of lesions related to this virus in the river communities of the target municipalities.
Acesso aberto (Open Access)
Prevalência da Hepatite B oculta nos pacientes atendidos no Núcleo de Medicina Tropical da Universidade Federal do Pará, Amazônia Brasileira
(Universidade Federal do Pará, 2016-03-15) SANT'ANNA, Carla de Castro; MARTINS, Luisa Caricio; http://lattes.cnpq.br/1799493244439769
Infection with hepatitis B occult (OBI) is characterized by absence of hepatitis B surface antigen (HBsAg) in commercial immunoassays, despite the persintência HBV DNA in the serum and / or liver tissue. The few studies of this clinical form in Brazil, mainly in the Amazon region, seen as an endemic area for hepatitis B virus (HBV), hamper the analysis to identify the epidemiological profile of the site. The objective of study was to determine the prevalence of hepatitis B occult in patients attended at the Tropical Medicine Center outpatient - Federal University of Pará, from January 2011 to December 2015, characterization of circulating viral genotypes and identify the main risk factors for the acquisition of this clinical form. Four hundred and sixty-five serum samples were submitted to ELISA (Enzyme-linked immunosorbent assay) for detection of serological markers of HBV (HBsAg, anti-HBc and anti-HBs). Total 181 patients resulted in a non-reactive HBsAg, anti-HBc reactive and anti-HBs nonreactive. These were screened with a PCR assay for identification of HBV-DNA. Subsequently, the positive samples were sequenced to identify the genotypes and mutations. Of the 181 samples, 26 (14,36%) had serum HBV-DNA, demonstrating the infection by OBI. The average age of 39 years, 53.45% (14/26) were married and 50% (13/26) were male. The genotype A was found in 88.46% (23/26), with the most prevalent subgenotype A1 with 78.26% (18/23) and genotype A2 with 21.73% (5/23). The genotype F was found in only 11.53% (3/26), in the presence of F2 genotype in all samples. As for risk factors only the nail pliers share was statistically significant. Some amino acid substitutions were identified in samples of patients with HBO compared with the HBsAg samples reagentes, but no mutation was identified. The study found a high prevalence of hepatitis B occult in patients treated at the Tropical Medicine Center – UFPA. HBV molecular studies are of fundamental importance for the identification of patients who are considered healthy, but they do have the infection and can be a transmitter disease potential.
Acesso aberto (Open Access)
Prevalência e variabilidade genotípica de Chlamydia trachomatis em amostras cervicais de estudantes universitárias em Belém, Pará, Brasil
(Universidade Federal do Pará, 2015) SANTOS, Leonardo Miranda dos; SOUSA, Maisa Silva de; http://lattes.cnpq.br/1775363180781218
Infection with Chlamydia trachomatis is a Sexually Transmitted Infection (STI) more prevalent bacterial in the world and can be asymptomatic in up to 80% of cases and is associated with late complications. The university students are part of a differentiated demand of the population for their high level of education. Objective was to assess the prevalence and variability of the genotypes of C. trachomatis in cervical infection of public university students in the state of Pará, Brazil, and to evaluate the association of the respective socio-behavioral characteristics and gynecological complaints. They included 438 college students from September 2012 to October 2014 and cervical samples were obtained during gynecological examination. It was performed phenol-chloroform technique for extracting total DNA from the sample cervical secretion, and for the detection of C. trachomatis used the Polymerase Chain Reaction (PCR) PCR-seminested the type for amplification 224pb gene omp1. For the identification of genotypes, we carried out a nested PCR for the amplification of the gene omp1 990pb, which was purified and subjected to ABI3130 sequencer, then the nucleotide sequences were compared with GenBank. The prevalence of cervical infection by C. trachomatis was 12.5% (CI: 95% ± 5.89) and genotypes were identified J genotype (36.3%), followed by D genotypes (18.2%) E (18.2%), F (18.2%) and la (9.1%). There was no significant association for age, first sexual intercourse, number of partners, and condoms are used, the presence of gynecological complaints and genotypes found in the study population. Although the prevalence found present itself high among university students, the lack of statistical significance may be due to small sample size and/or result in socially acceptable answers. Efforts are made to trace the expansion of C. trachomatis infection in restricted populations.
Acesso aberto (Open Access)
Reconstrução e modelagem in silico da via de biossíntese de ácidos graxos da bactéria psicotrófica Exiguobacterium antarticum linhagem B7
(Universidade Federal do Pará, 2016-04-04) FRANCÊS, Regiane Silva Kawasaki; SCHNEIDER, Maria Paula Cruz; http://lattes.cnpq.br/3901112943859155
Mathematical modeling in silico based restrictions is an approach adopted by systems biology to analyze metabolic networks. The Gram-positive bacterium Exiguobacterium antarticum B7 is an extremophile organism able to survive in cold environments as glacial ice and permafrost. The ability of these microorganisms of adaptation to cold attracts great biotechnological interest. An important factor for the understanding of cold adaptation process is related to the chemical modification of fatty acids constituting the cell membrane of psicotrophic bacteria in order to maintain membrane fluidity to avoid freezing ofthe bacteria. In this work, the metabolic pathway of fatty acid biosynthesis of the bacterium E. antarticum B7 was rebuilt from its annotated genome. The software tools KEGG (Kyoto Encyclopedia of Genes and Genomes) and RAST (The Rapid Annotation Server) were used to generate a preliminary network model. The next step was to cure manually the genomic, biochemical and physiological informations available in different databases and specific literature. During this process, the FabZ and DesK enzymes responsible for adding carbon-carbon unsaturations in the fatty acid chain during synthesis have been identified in the genome, though in a truncated form. The fluxome metabolic pathway was defined, describing the routes of the main reactions since the first monomer, Acetyl-CoA, to the final product, the Hexadecenoic acid. A computational modeling was done using the software MATLAB® with toolboxes and specific tools for systems biology. The quantification of metabolites produced via was performed by the method constraint-based Flux Balance Analysis (FBA). To evaluate the influence of the gene expression in the fluxome analysis, the FBA method was also calculated using the log2FC values obtained in the transcriptome analysis at 0ºC and 37ºC. The fatty acid biosynthesis pathway showed a total of 13 elementary flux modes, four of which showed routes for the production of hexadecenoic acid. The reconstructed pathway demonstrated the capacity of E. antarcticum B7 to produce fatty acid molecules. Under the influence of the transcriptome, the fluxome was altered, promoting the production of short-chain fatty acids. The calculated models contributes to better understand the bacterial adaptation at cold environments.