Navegando por CNPq "CNPQ::CIENCIAS BIOLOGICAS::GENETICA::GENETICA MOLECULAR E DE MICROORGANISMOS"

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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/8620752020290438
Mycobacterial 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.
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Análise da expressão de miRNAs em carcinoma hepatocelular
(Universidade Federal do Pará, 2014-02-11) SANTOS, Ian Barroso dos; DEMACHKI, Samia; SANTOS, Ney Pereira Carneiro dos; http://lattes.cnpq.br/7568391537270652; http://lattes.cnpq.br/1290427033107137
Hepatocellular carcinoma represents the most common primary malignancy of the liver and the fifth most common solid tumor worldwide. Highly lethal, remais a serious public health problem because of difficulties in early diagnosis and the development of effective therapeutic measures. Recent in the field of molecular biology studies suggest that define the profile of miRNAs in hepatocellular carcinoma may considerably influence the identification of risk factors associated with oncogenes and suppressor genes. The objective is to evaluate the expression of miRNA 135b, miRNA 181a-5p and miRNA 181a-3p in samples of Hepatocellular Carcinoma and Chronic Hepatitis C and correlate them so likely to seek biomarkers related to the mechanism of carcinogenesis. The research was done in six patients with hepatocellular carcinoma and twenty four cases of Chronic Hepatitis C, Para, northen Brazil. All samples Hepatocellular carcinoma underwent microdissection for subsequent RNA extraction. For the extraction of total RNA and microRNA All=Prep the DNA / RNA FFPE kit (quiagem), quantified by the Qubit® 2.0 Fluorometer (Invitrogen) for final concentration of 5ng/μL standard equipment was used. The cDNA was obtained using TaqMan® MicroRNA Reverse Transcription (Applied Biosystems). Statistical analyzes were performed in softwares SPSS 17.0, using the Mann-Whitney test, with significat differences in the expression levels of the miR181a-3p and miR 181a-5p in hepatocellular carcinoma (average 3.94 and 17.9, respectively) compared with chronic hepatitis C (average 1.18 to 1.8, respectively) with P-value of 0.005 and 0.003. In this study, it was observed that miRNAs 181a-3p and 181a-5p, especially the 181a-5p way were significantly more highly expressed in hepatocellular carcinoma samples when compared to non-tumor liver tissue with chronic hepatitis C. Therefore, microRNAs have interesting characteristics that favor them as possible in biological screening for early diagnosis of tumors and targeted therapies selected markers.
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Análise da expressão gênica de pacientes com adenocarcinoma gástrico associado ao vírus Epstein-Barr
(Universidade Federal do Pará, 2024-08) SILVA, Valéria Cristiane Santos da; MOREIRA, Fabiano Cordeiro; http://lattes.cnpq.br/5745396559731337
Infection with the Epstein-Barr virus (EBV) is one of the risk factors for gastric cancer (GC). Epstein-Barr is a virus with oncogenic activity and was the first to be associated with malignant diseases such as lymphomas and carcinomas. Thus, EBV detection can be performed both by in situ hybridization and, currently, by using RNA sequencing techniques to identify the presence of viral genes in samples. In this context, aiming to better understand the EBV-positive subtype, this study proposed a molecular classification based on RNA sequencing of 76 tumor samples from patients diagnosed with GC. RNA sequencing of the samples was performed, and molecular classification was done using the Kraken2 software. Of the 76 samples, 8 were considered positive according to the adopted method. Subsequently, to understand the different mechanisms by which EBV might be acting in gastric cancer, we analyzed the patterns of human gene expression in samples classified as positive and negative. In our study, there are approximately 834 differentially expressed genes, of which 92 have an AUC greater than 0.85. These genes are associated with tumor progression, cellular metabolism, and innate and adaptive immunity. Additionally, viral genes expressed in the positive samples were evaluated, and we found manifestations of both lytic phase and latent phase genes. Finally, our study presents an efficient strategy for the molecular classification of EBV-positive
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Análise de polimorfismo na região promotora do gene da Interleucina 18 (-137 G/C e -607 C/A) em pacientes portadores do vírus da hepatite C de Belém, Pará
(Universidade Federal do Pará, 2012-03-23) SANTOS, Kemper Nunes dos; MARTINS, Luisa Caricio; http://lattes.cnpq.br/1799493244439769
Since its discovery in 1989, the hepatitis C virus (HCV) has been recognized as a major cause of chronic liver disease worldwide. Considered a public health problem worldwide involving between 170 to 350 million people infected. Host genetic factors have been implicated in the persistence of HCV infection. Studies suggest that two single nucleotide polymorphisms at position -607 C/A (rs1946518) and -137 G/C (rs187238) in the region of the gene IL-18 and have been found associated with the transcriptional promoter activity of IL -18, and potentially of IFN-γ, being associated with delayed viral clearance and persistence of the disease. We conducted a cross-sectional analytical study of the city of Belém-PA in 152 blood samples from patients infected with HCV and 188 uninfected controls. The samples were subjected to RT-PCR (Reverse Transcriptase - PCR) for detection of viral RNA and, subsequently, the PCR-RFLP (Restriction Fragment Length Polymorphism) to evaluate the polymorphism in the promoter region of IL-18 gene at positions -137 G/C and -607 C/A. The results showed no significant difference for IL-18 polymorphisms between patients and control group. But showed a significant difference for homozygous genotypes G/G (39.1%) at position -137 (OR = 3.00, CI [95%] = 1.24 – 7.22, p = 0.02), and A/A (21.7 %), position -607 (OR = 3.62, CI [95%] = 1.25 – 10.45, p = 0.03) among women than men (22.6% and 7.6%). The results showed evidence that among women, the presence of the polymorphism homozygous A/A (-607) acts as a protective factor against HCV infection, genotype as the A/A (-607) have been linked in some studies with liver disease and mild viral clearance.
Acesso aberto (Open Access)
Análise epidemiológica e caracterização parcial do gene F dos Metapneumovirus Humano detectados a partir de casos de infecção respiratória aguda na Região Norte do Brasil
(Universidade Federal do Pará, 2012-10-30) FERREIRA, Luís Edilson de Azevedo; MELLO, Wyller Alencar de; http://lattes.cnpq.br/1784167608719139; SOUSA, Rita Catarina Medeiros; http://lattes.cnpq.br/3560941703812539
Diseases of the respiratory tract are the main complaints in health care services, and acute respiratory infections (ARI) the most common manifestations, especially in children under five years of age. In developing countries the ARI are a serious public health problem. Throughout the world is estimated to occur in approximately 2 million deaths each year by ARI. Among the causative agents of ARI, we highlight the human metapneumovirus (HMPV), especially by causing severe illness in children under 5 years. Aiming to generate data on the molecular epidemiology of this virus, we analyzed samples taken from patients with ARI in the period January 2009 to December 2011 arising from the Northern Region (Pará, Amazonas, Acre, Amapá and Roraima). It was used the technique of real time PCR (qRT-PCR) to detect the virus by the N gene and RT-PCR for the gene encoding the F protein, which was then partially sequenced. Within the study period, were tested in 2966 samples, 129 of which were positive for HMPV. The age group of 0-4 years was the one that focused more cases (n = 84, 65.89%) across the North. The identification of viruses was found to co-circulation of subgroups A2 and B2 during the three years of the study. Subgroups A1 and B1 not circulated in the region during the study period. This study represents the first report of data on molecular epidemiology of Human Metapneumovirus in northern Brazil.
Acesso aberto (Open Access)
Análise filogenética de genes de provável origem não humana de rotavírus do grupo A em espécimes fecais de crianças com gastrenterite aguda provenientes de Belém, Brasil
(Universidade Federal do Pará, 2012) MAESTRI, Régis Piloni; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Rotaviruses (RVs) are the main cause of acute viral gastroenteritis in both humans and young animals of species such as calves, horses, pigs, dogs, cats, and birds. The genetic diversity of RVs is related to a variety of evolutionary mechanisms, including point mutation, genome reassortment, and reassortment. The objective of this study was realized the molecular charaxterization of the genes that encode structural and nonstructural proteins in unusual RV strains. The clinical specimens selected for this study were obtained from children and newborns with RV gastroenteritis, who participated in research projects on viral gastroenteritis conducted at the Evandro Chagas Institute. Structural (VP1-VP4, VP6, and VP7) and nonstructural (NSP1-NSP6) genes were amplified from stool samples by the polymerase chain reaction and subsequently sequenced. Eight unusual RV strains isolated from children and newborns with gastroenteritis were studied. Reassortments between genes of animal origin were observed in 5/8 (62.5%) strains analyzed. These results demonstrate that, although rare, interspecies (animal-human) transmission of RVs occurs in nature, as observed in the present study in strains NB150, HSP034, HSP180, HST327, and RV10109. This study is the first of its kind conducted in the Amazon region and supports previous data showing a close relationship between genes of human and animal origin, representing a challenge to the large-scale introduction of RV vaccines in national immunization programs.
Acesso aberto (Open Access)
Análise molecular de rotavírus tipo G9 de crianças na Região Norte do Brasil
(Universidade Federal do Pará, 2016-10-31) GUERRA, Sylvia de Fátima dos Santos; SOARES, Luana da Silva; http://lattes.cnpq.br/0556695301015859; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Group A rotavirus (RVA) is the most viral agent associated with acute gastroenteritis, responsible for about 200,000 deaths among children aged under five years annually. RVA belongs to Reoviridae family, Rotavirus genus, its genome is composed by double-stranded RNA (dsRNA) with 11 segments encoding 12 proteins, six structural (VPs) and six non-structural (NSPs). Each protein designating a specific RVA genotype, being VP7 protein responsible for G genotype and currently there are 32 genetic variants. G9 genotype emerged on a global scale in the 90s, a period before RVA vaccine introduction in Brazil that occurred in 2006, and is continuously detected until present day. This study aimed to describe the frequency and genetic constellation associated with the current G9 genotype in Northern Brazil. It was selected 50 samples collected between 1999 and 2013, being 45 G9P[8], 2 G9P[4] and 3G9P[6], for fecal suspension preparation and dsRNA extraction for further genome amplification and sequencing of nucleotides. It was observed that during pre-RVA vaccine introduction period G9 frequency rate was 43%, while after RVA vaccine introduction the most frequece obtained was 12.5% (2008 to 2010). Phylogenetic analysis of VP7 gene showed that all strains belong to lineage III of G9, observing aminoacidic substitutions in antigenic sites when compared with vaccine strains. It was demonstrated in VP4 gene that P[8] strains gathered in lineage III, whereas P[4] grouped into lineage V and P[6] strains into lineage I. All G9P[6] and G9P[4] samples were associated with DS-1 constellation, genogroup 2, while G9P[8] samples showed Wa constellation, genogroup 1, except for one sample showing NSP3 gene with DS-1 profile. G9 samples from Northern region analyzed were associated with the expected constellations described in other parts of the world, except for one G9P[8] sample that showed a genetic restructuration in NSP3 protein. In the present study the same G9 lineages have circulated during pre and post RVA vaccine introduction periods, and it was described aminoacidic substitutions in relevant antigenic regions, such as it was reported genetic restructuration phenomenon in one sample of this genotype, emphasizing the continuous monitoring of current genetic variants of RVA.
Acesso aberto (Open Access)
Avaliação do perfil de metilação e expressão do gene CDH1 em Cebus apella como modelo experimental para câncer gástrico
(Universidade Federal do Pará, 2012-07-09) ANTUNES, Symara Rodrigues; BORGES, Barbara do Nascimento; http://lattes.cnpq.br/0676220027193876; ANSELMO, Nilson Praia; http://lattes.cnpq.br/6518287721873199
The gastric cancer remains a major cause of death among cancers in the world. In Brazil, are expected around 500 000 new cases in 2012/2013. The origin of stomach cancer, as in others, arises from the accumulation of genetic alterations. Therefore, it is necessary to know which genetic changes are important in triggering the pathogenesis of gastric cancer. We know that the intestinal type develops through well defined stages. The MNU - a known carcinogen - when ingested in oral form at dosages determined triggers development of this histological type of gastric cancer. Based on this knowledge, we conducted an experiment with six specime of Cebus paella, induced to develop intestinal type gastric cancer. The animals consumed 16mg/kg of body weight daily drugs. All pre-neoplasic lesions developed. Due to drug toxicity, only one survived the entire treatment and developed a tumor. Periodic evaluations were made of animals in pre-determined days (the beginning, days 120, 150, 300, 940) during which samples were made of the gastric mucosa. We collected 20 samples of tissue, distributed among normal mucosa, gastritis, dysplasia, metaplasia, and tumor. DNA extracted from these samples for further analyzes of the CDH1 gene. There is no sequence of this gene in the literature for the species under study. So the first step was to get this sequence. Using the initiators constructed from sequences of CDH1 Callithrix jacchus (species phylogenetically closer to C. apella) we get a sequence of 342pb. There is a similarity of 98% with humans, the presence of binding sites for transcription factors like (sp1, Ap2, NF-x, AREB6, Puf and CTF) and the presence of CAAT Box. The sequence has 30 CpG sites could suffer epigenetic regulation. We also performed MSP analysis with specific primers and found that the region analyzed, there was predominance of the unmethylated CDH1 alleles for all samples pre-neoplastic and adenocarcinoma sample is methylated. When we compared these data with immunohistochemistry revealed that only tumor sample did not express the protein cadherin. Our analyzes suggest that methylation of the CDH1 gene plays an important role in gastric tumorigenesis in C. apella. And, from the similarity between sequences, we suggest that the same occurs in humans. This cadherin promoter methylation leads to inactivation of the gene and establishment of gastric adenocarcinomas. The cadherin promoter hypermethylation has been reported in several articles associated with the development of gastric cancer.
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/1274395392752454
The 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.
Acesso aberto (Open Access)
Avaliação epidemiológica, clínica e molecular de enteropatógenos causadores de diarreia aguda em crianças e adultos residentes na comunidade Quilombola do Abacatal, Ananindeua, Pará
(Universidade Federal do Pará, 2015) KAIANO, Jane Haruko Lima; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435
Acute diarrheal disease is a major cause of morbidity and mortality in developing countries and one of the factors that contributes to the worsening of the nutritional status of children. This study aimed to evaluate the clinical, epidemiological and molecular profile of infections by viral and parasitic agents in children aged 0-10 years and those over 10 years of quilombo of Abacatal in the 2008-2010 period. Fecal samples from 294 children were collected in the age group 0-10 years and 81 individuals over 10 years, residents of the community Abacatal, Ananindeua, Pará, which had acute diarrhea board or without diarrhea (controls). The viral diagnosis was made by immunochromatographic and molecular tests and parasitological by Faust and Hoffman method. A total of 375 fecal samples were obtained from 177 individuals. The frequency of viral agents in this study were rotavirus group A rotavirus C and picobirnavirus group by 6.4% (24/375), 0.3% (1/375) and 1.3% (5/375 ), respectively.The polyacrylamide gel electrophoresis confirmed the presence of rotavirus in 23 of 10 samples (43.48%) having short profile of 13 (56.5%) long profile. The presence of intestinal parasites was observed in 272 (77.94%) samples, and the most common were Ascaris lumbricoides detected in 13.18% (46/349) of the samples, followed by Trichuris trichiura with 10.88% (38 / 349), hookworms with 4.01% (14/349) and Strongyloides stercoralis 1.72% (6/349). Of the 24 samples positive for rotavirus group A the following genotypes were detected: G2P [4] (12.50%, 3/24); G1P [8] (25.00%, 6/24), G3P [9] (29.20%, 7/24) and G12P [6] (33.33%, 8/24). Two new genotypes were detected for VP6 genes (I18) and NSP1 (A19) of rotavirus A. Nutritional assessment of 38 children was conducted, showing that 18 4% (7/38) presented malnourished. This study highlights the need to implement preventive actions in the community, including education measures for health, vaccination against rotavirus, and even the implementation of programs to control parasitic infestations.
Acesso aberto (Open Access)
Características epidemiológicas relativas à doença dental e infecção por Helicobacter pylori na cavidade oral de estudantes em Belém-Pará
(Universidade Federal do Pará, 2009) MATOS, Gyselly de Cássia Bastos de; CORVELO, Tereza Cristina de Oliveira; http://lattes.cnpq.br/7253864056606024
The infection by Helicobacter pylori is one of the most common in humans and despite having tropism by stomach, can be found in the oral cavity, maintaining a commensal relationship with the host, while dental caries is also an infectious disease and results from the metabolism of the bacterial plaque. Both infections are highly prevalent in developing countries, since these populations are more exposed to environmental risk factors, and are usually acquired during childhood. The prevalence of these infections was investigated in the oral cavity of school children with no symptoms of gastric diseases, from a population of Belém-Pa, in relation to some parameters of oral hygiene and health, socioeconomic conditions and genetic susceptibility factors like the ABO and Lewis blood groups. Were investigated 104 patients, with average age of 17 years. Of all the participants were collected saliva samples and dental plaque. Saliva was collected to identify the ABO and Lewis state secretor and estimation of salivary parameters, and both, saliva and plaque samples were collected for molecular analysis of 16S rRNA genes of H. pylori and FUT2. H. pylori was detected in 79.8% of the students, with a frequency of 66.35% in dental plaque and 58.65% in saliva. The caries prevalence was 82.8% in the population studied. The clinical evaluation of oral health showed that the average CPO-D found was 3.53. It was observed that the caries experience tends to increase as in addition to age and the H. pylori infection was higher in early childhood. The education level and number of dentist visits showed significant differences in relation to the presence of H. pylori. The phenotypic distribution of ABO and Lewis blood groups did not differ significantly between infected and uninfected individuals, explaining there is greater genetic susceptibility to infection by H. pylori and dental caries. Throughout this analysis, the high frequencies found prove the need for care and treatment of dental diseases, such as caries and it is suggested that H. pylori in the oral cavity can contribute to infection and re-infection of the stomach after treatment.
Acesso aberto (Open Access)
Caracterização antigênica e molecular de amostras de rotavírus do tipo G1, obtidas de crianças participantes de estudos em gastroenterites virais, no período de 1982 a 2003, em Belém, Pará, Brasil
(Universidade Federal do Pará, 2006-07-03) SOARES, Luana da Silva; MASCARENHAS, Joana D'Arc Pereira; http://lattes.cnpq.br/5156164089432435; LINHARES, Alexandre da Costa; http://lattes.cnpq.br/3316632173870389
Infant mortality remains an important problem of public health worldwide, mainly in developing countries. Of more than the 50 etiologic agents implied in this disease, rotavirus causes 111 million episodes of diarrhoea, resulting in more than 600,000 deaths among children less than five years, of which 82% are notified in the poorest countries of the world. This study aimed at the antigenic and molecular characterization of G1 rotavirus strains among children participanting of viral gastroenteritis studies, carried out from 1982 to 2003, in Belém, Pará, Brazil. One hundred and forty-eight specimens of G1 rotavirus were analyzed in the present investigation. Overall, the prevalence of the G1 type was of 41.3%, being that frequencies of this genotype through studies ranged from 11.0% to 67.6%. Eletropherotypes, G serotypes and P genotypes characterization of G1 rotavirus occurred in frequencies of 78.4%, 89.9% and 87.8%, respectively. Three long eletropherotypes varieties were identified, being that the L1 variety was found frequently (79.3%). The G1, G9 and G1+G4 serotypes were detected in 88.0%, 9.8% and 2.2% of the specimens, respectively. Mixed infection by G1+G4 genotype was detected in one sample. The prevalent binary combination was P[8],G1, being responsible for 72.3% of the cases. Mixed infections circulated in percentage of 20.0%, including genotypes P[4]+P[8],G1, P[6]+P[8],G1, P[4]+P[6],G1, P[4]+P[6]+P[8],G1 and P[6]+P[8],G1+G4. The G1 genotype circulated among 2nd to 35th months of age and a highest number of cases was registered between 6 to 16 months of age. Clinical severity differences among G1 and other genotypes of rotavirus were not verified. The present analysis gathers pioneer findings in Brazil, allowing to extend the knowledge concerning the antigenic and molecular diversity of the infections by G1 rotavirus and these results will allow to understand the genetic complexity of such viral agents.
Acesso aberto (Open Access)
Caracterização de espécies de Leishmania isoladas de Flebotomíneos sp. de três ecótopos da Serra dos Carajás, Pará, Brasil
(Universidade Federal do Pará, 2008) DIAS JUNIOR, Manoel Guacelis de Sena; ISHIKAWA, Edna Aoba Yassui; http://lattes.cnpq.br/3074963539505872
The American cutaneous leishmaniasis (LTA) is an infectious disease, with a high frequency of zoonosis, endemic in the Amazon region, transmitted by sand flies of generous Psychodopygus and Lutzomyia. Serra dos Carajás, located in southeastern Pará, is widely exploited by extractive companies and as a result, the LTA would be transformed into a major health problem to workers because of the practice of deforestation and construction of roads for drainage of the ore. The purpose of this study is to evaluate the natural infection in the sand fly by Leishmania in the Serra dos Carajás region through the PCR. Catches of sandflies were held in three different areas, Parque Zoobotânico de Quarentena, APA do Gelado and Tapirapé-Aquirí, with CDC-type of light traps and Shannon, during the night from the evening twilight. The sand flies captured were identified according to Young & Duncan, 1994, and frozen in N2. 5.947 sandflies were frozen, being 3.495 females and among these 550 specimens were tested. The extraction of DNA was performed using SDS and KOAc and it was precipitated with ethanol 96%. Subsequently, the PCR was performed by amplifying the mini-exon gene with the primers S1629 (5' GGGAATTCAATAWAGTACAGAAACTG 3') and S1630 (5' GGGAAGCTTCTGTACTWTATTGGTA 3'). The DNA of Leishmania was detected in 36 (6,5%) sand flies, 34 of subgenus Viannia detected in 30 Psychodopygus wellcomei/ complexus, three Lutzomyia whitmani and a Lutzomyia shawi. Two infections by Leishmania amazonensis was detected in Psychodopygus wellcomei/ complexus. Tapirapé - Aquirí, APA do Gelado and Parque Zoobotânico de Quarentena showed high rates of natural infection in sand flies 6.54%, 5.96% and 7.92% respectively. Ps. wellcomei/complexus still presents as Leishmania vector that cause LTA. Studies on the power of vector species Lu. whitmani and Lu. shawi naturally infected by Leishmania in the Serra dos Carajás should be intensified, if these species may be acting in the cycle of transmission of LTA in Serra dos Carajás. Studies that best explain the variation of the prevalence of different sandflies species and knowledge of the rate of infections should be also intensified in Serra dos Carajás.
Acesso aberto (Open Access)
Caracterização dos genes codificadores da hemaglutinina e PB2 do vírus Influenza A (H1N1) pandêmico isolado na mesorregião metropolitana de Belém
(Universidade Federal do Pará, 2012-10-26) FERREIRA, Jessylene de Almeida; SOUSA, Rita Catarina Medeiros; http://lattes.cnpq.br/3560941703812539; NUNES, Márcio Roberto Teixeira; http://lattes.cnpq.br/0299116892743368
The recent influenza pandemic of 2009/2010 caused by the Influenzavirus A (H1N1) pandemic showed a severity profile different from seasonal flu due to a significant percentage of severe and fatal cases occurred in young adults without comorbidity. The virulence of Influenzavirus A (H1N1) pandemic is the result of protein interaction complexes and is related essentially some viral genes. The aim of this study was to characterize the genes that encodes for the hemagglutinin (H1) and polymerase basic 2 (PB2) of Influenzavirus A (H1N1) pandemic recovered from patients with flu coming from the metropolitan mesoregion, Belém-PA. The sample size consisted of 87 random samples of both genders, the 0-96 years, with severe acute respiratory syndrome (SARS) without comorbidity reported from May 2009 to August 2010. The samples were isolated in MDCK cell, and analyzed by molecular biology techniques that comprised three main steps: a) viral RNA (vRNA) extraction from supernatant of infected cells; b) amplification of the vRNA by Polymerase Chain Reaction preceded by Reverse Transcription (RT-PCR) technique; c) complete sequencing of genes encoding H1 and PB2. Of 87 strains amplified by RT-PCR in 82 amplicons the acquisition and analysis of sequences for the HA gene was obtained, whereas in 81 amplicons viral sequences were obtained for the PB2 gene. The comparative analysis of the sequences obtained with the sequence of the vaccine strain (A/California/07/2009 (H1N1)) revealed amino acid substitutions in HA (P83S, D97N; S203T, D222G, and I321V Q293H) and PB2 (K340N, and K526R M631L) proteins any changes were, however not associated with hospitalization. At the level of substitution in HA, the D97N alone or associated with the S203T was detected more frequently in the first wave. Furthermore, the level of PB2, a substitution K526R was found in the majority of strains that circulated during the first wave, while the M631L was more evident in the second. The D222G substitution in HA was only found in cases of death. Finally, there was a tendency of changes in HA antigenic sites. Thus, the genetic and antigenic continuous surveillance of Influenzavirus A (H1N1) pdm in circulation, as well as the sharing of information is extremely important for the best possible recommendation for virus which are included in vaccine the composition thus avoiding higher risk of severe epidemics in the future.
Acesso aberto (Open Access)
Caracterização dos genótipos do vírus da hepatite b em pacientes atendidos no programa de hepatites virais do núcleo de medicina tropical – UFPA, Belém - Pará
(Universidade Federal do Pará, 2016-05-27) ALMEIDA, Marcella Kelly Costa de; MARTINS, Luisa Caricio; http://lattes.cnpq.br/1799493244439769
The hepatitis B virus belongs to the genus Orthohepdnavirus and Hepadnaviridae family, comprising a DNA virus capable of infecting mammalian hepatotropic. Classified into 10 genotypes (A-J) and many different subgenotypes, studies suggest that they may influence the severity of the disease, response to treatment and vaccine response. Genotypes and subgenotypes HBV has a wide distribution, some being restricted to certain geographic regions, while others show a worldwide distribution. It is found in different regions of Brazil with prevalence of genotypes A, D and F. This study aimed to identify genotypes and subgenotypes of hepatitis B virus among patients treated at the Tropical Medicine Center - UFPA, in the city of Belém, state Pará, from January 2011 to December 2015. A total of 1274 patients treated at the NMT-UFPA in the period, were selected for the study 222 patients of both sexes, with positive serology for HBsAg and / or anti-HBc. The samples were subjected to molecular biology, PCR "in house" and multiplex PCR to detect viral DNA and genotyping, and then the sequencing for confirmation and determination of viral subgenotypes. In 65 of the 222 samples was detected the presence of HBV DNA, but only 63 samples were satisfactory after the result of sequencing, which were identified the presence of genotypes A (46/63), especially the subgenotype A1 (36/46) followed by subgenotype A2 (10/46), and genotype F (17/63) and only detected subgenotype F2 (17/17) circulating in this population. The average age of the patients was 38 years, predominantly male, and most natural state of Pará patients. Some risk factors have been identified among the population studied, and not using a condom during sex the most predominant. Comparing the presence of viral DNA with serology for HBsAg marker is shown a picture that suggests the presence of occult hepatitis among the patients treated. The results found in this study are consistent with what is reported in other studies in Brazil.
Acesso aberto (Open Access)
Caracterização genética parcial e completa da nucleoproteína de hantavírus na Amazônia brasileira
(Universidade Federal do Pará, 2011) SIMITH, Darlene de Brito; NUNES, Márcio Roberto Teixeira; http://lattes.cnpq.br/0299116892743368
The Hantavirus Pulmonary Syndrome (HPS) has been diagnosed in the Brazilian Amazon since 1995. Until december 2010 have been diagnosed 289 cases in the Brazilian Amazon, registered in the states of Mato Grosso, Pará, Maranhão, Amazonas and Rondônia. The overall objective of this study was to characterize genetically hantavirus strains circulating in these states. Samples of viscera from wild rodents positive for IgG antibodies against hantavirus caught in ecoepidemiológicos studies, conducted in the municipalities of Itacoatiara/AM, Alto Paraíso/RO and Campo Novo do Parecis/MT, and serum/blood of human cases of HPS from the municipalities in the area of influence of BR-163 in the states of Pará and Mato Grosso, Tomé-Açu/PA, Tangará da Serra/MT, and viscera of a pool of death coming from Anajatuba/MA. The samples were extracted viral RNA, followed by the reactions of RT-Hemi-Nested-PCR for samples from rodents, RT-Nested-PCR for human samples and nucleotide sequencing using the Sanger method and pyrosequencing, and later, scanned for matters such as identity (BLAST search), similarity (Simplot) and nucleotide and aminoacidic homology with other hantaviruses (Clustal W). We obtained partial sequences of hantavirus in five species of rodents Oligoryzomys microtis (n=2 from Itacoatiara/AM; n=3 from Alto Paraíso/RO) and in eight samples from humans (n=1 from Tomé-Açu/PA; n=1 from Altamira/Cachoeira da Serra; n=1 from Novo Progresso/PA; n=1 from Guarantã do Norte/MT; n=1 de Anajatuba/MA and n=3 de Altamira/Castelo dos Sonhos). Using the strategy of pyrosequencing were obtained complete sequences of the gene N, S-RNA of three hantavirus in rodents (n=2 from Alto Paraíso/RO and n=1 from Campo Novo do Parecis/MT) and two human cases (n=1 from Tangará da Serra/MT and n=1 from Novo Progresso/PA). Analysis of complete sequences showed the presence of ORFs for possible NSs protein, as described for other hantaviruses. Phylogenetic analysis of the sequences obtained in this study and other hantaviruses available in GenBank suggests that the virus Castelo dos Sonhos is responsible for cases of HPS in municipalities in the area of influence of BR-163, obtaining for the first time, the complete sequence of this virus in rodent Oligoryzomys utiaritensis, coming from Mato Grosso; confirmed the continued circulation of Laguna Negra-like virus associated with HPS cases in the state of Mato Grosso; the Rio Mamoré-like virus was first time detected in O.microtis rodents, the state of Amazonas and Rondônia, but not associated with human cases; the virus Anajatuba was responsible for a case of death from Maranhão. This work will serve as support for future epidemiological and molecular studies, therefore, provides new data about the spread of hantaviruses in the Brazilian Amazon.
Acesso aberto (Open Access)
Caracterização molecular dos vírus do grupo Gamboa (Bunyaviridae, Orthobunyavirus) isolados nas américas e infecção experimental em pintos (Gallus gallus domesticus) com o vírus Gamboa cepa Be AN 439546
(Universidade Federal do Pará, 2010-03-31) CHIANG, Jannifer Oliveira; VASCONCELOS, Pedro Fernando da Costa; http://lattes.cnpq.br/0973550817356564
Presently, little information on Gamboa serogroup viruses (Bunyaviridae, Orthobunyavirus) is available. Thus, in this work, it was performed a comparative phylogenetic study on the members of the Gamboa serogroup and with other orthobunyaviruses to the level of the gene Gn (M-RNA); an experimental infections in the domestic bird (Gallus domesticus) using the strain Be AN 439546 of the Gamboa Virus (GAMV); and a serologic study using the Hemagglutination Inhibition (HI) test in serum samples of wild animals and humans collected in Tucuruí - Pará. The phylogenetic analysis of Gamboa group viruses demonstrated that they are genetically closely related to group Turlock viruses and less related to the Simbu group viruses. The group Gamboa viruses were distributed in two clades (I and II), that it is in agreement with the current serologic classification; the clade I correspond to the Gamboa complex and the clade II to the Alajuela complex. The strain Be AN 439546 presented tropism for chikens lung and liver, with viral replication in this organs confirmed by detection of viral antigens by immunohistochemistry. These results, demonstrate that this bird species is a susceptible host for GAMV replication. The detection of HI antibodies against GAMV, confirmed by neutralization tests were found in wild bird plasmas and reinforces the hypothesis that these animals constitute the main amplification hosts in the maintenance cycle of GAMV. Full length genome studies of the Gamboa serogroup viruses, as well as on the ecoepidemiology of their vectors and potential vertebrate hosts are needed to generate new data and to reinforce the understanding of the information already existent on those viruses.
Acesso aberto (Open Access)
Caracterização sorológica e detecção molecular do HTLV em amostras de pacientes com distúrbios neurológicos no Estado do Pará, Brasil (1996-2005)
(Universidade Federal do Pará, 2006-07-07) LIMA, Telma Vitorina Ribeiro; LINHARES, Alexandre da Costa; http://lattes.cnpq.br/3316632173870389
Human T-lymphotropic virus tipe 1 is recognized as the etiologic agent of tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM). A very similar clinical disease has been increasingly associated to HTLV-2, whose pathogenicity still requires further assessments. This transversal, retrospective epidemiological survey aimed to determine the prevalence of HTLV among individuals with neurological disturbances and further evaluate cases of inconclusive serology using molecular biology methods. The present study involved patients inhabitants of Pará State and/or admitted at health institutions of the and who were referred to the Virology Section of Instituto Evandro Chagas (IEC) by local doctors between January of 1996 and December 2005, to search for the presence of HTLV-1/2 serum antibodies. Of these patients 353 were selected, with age between 9 months and 79 years, who presented at least one signal or symptom of the Marsh’s Complex (1996), as well as had HTLV-1/2 positive serology at screening and confirmatory ELISA. The overall prevalence of HTLV antibodies by ELISA as 8,8% (31/353), with rates of 10,6% (19/179) and 6,9% (12/174) for female and male patients, respectively. Among HTLV-1/2 the 31 ELISA-positive patients it was noted that 15 (48.4%) of 31 had paresis (n = 8), parestesis (n = 5), and paraplegia (n = 3). Of these 31 HTLV ELISA positive patients, 25 could be submitted to WB for assessment of viral types, which were distributed as follow: 80% (20/25) were HTLV-1, 12% (3/25) were HTLV-2, one case was of HTLV-1+HTLV-2 infection (4%), and serum from one patient yielded an indeterminate profile (4%). Only 14 of these 25 patients could be re-localised for collection of an additional sample for molecular analysis. It was observed that 78.6% of samples typed by WB had the proviral TAX region successfully amplified by nested-PCR. In addition, types were confirmed as based on results obtained from the amplification of the POL region using real-time PCR; this denoted good specificity and sensitivity of the WB used in this study. The sample defined as HTLV-1+HTLV-2 infection by WB was amplified in its TAX region but real time PCR confirmed HTLV-1 infection only. The patient with WB indeterminate profile and one of samples typed as HTLV-2 by WB were amplified by nested-PCR but the real time PCR was negative for HTLV-1 and HTLV-2 in both samples. One patient presenting clinical manifestations of crural myalgia and parestesia with duration of about 7 years reacted HTLV-2-positive by both WB and real-time PCR, a denoting a clear HTLV-2- related chronic myelopathy. This study has identified a case of possible vertical transmission in two distinct situations: a patient whose mother presented antibodies for HTLV-1 by WB and two sisters who reacted HTLV-1-positive by WB and real-time PCR. Although of epidemiological relevance, results from this study warrant further and broader analyses concerning the molecular epidemiology of HTLV types and subtypes HTLV. In addition, a more complete clinical assessment of neurological symptoms should be further performed, in order to better characterise cases of HTLV-related chronic myelopathy in our region.
Acesso aberto (Open Access)
“Citomegalovírus: diversidade genética e pesquisa de resistência antiviral em pacientes imunodeficientes da cidade de Belém”
(Universidade Federal do Pará, 2015-07-17) SILVA, Dorotéa de Fátima Lobato da; SOUSA, Rita Catarina Medeiros; http://lattes.cnpq.br/3560941703812539
Cytomegalovirus (CMV) is one of the most common cause of morbidity and mortality in immunocompromised patients because its latency and reactivation mechanism that commonly occurs in immunodeficiencies. Genetic analysis showed that the virulence of the strains may be related to genotypic diversity. The main objective of this paper was to describe the seroepidemiology profile and genetic diversity of CMV by detecting mutations that confer viral resistance to ganciclovir in immunodeficient patients from Belém city.A total of 671 samples were analyzed: 243 HIV/AIDS, 257 neoplastic patients, 112 kidney transplant and 60 people with SLE. The seroprevalence of antibodies was 96.1% and active infection and levels of 2.4% (n = 16) lower than that observed by qPCR method which corresponded to 15.63%. Differences in infection rates due to low sensitivity (5.71%) of the serological method demonstrated in screening test. The mutation research was made in 82 samples for pyrosequencing method, a 741pb segment of the UL97 gene was amplified, between 1087-1828 nucleotides. It was observed that 100% (n = 82) of samples had two mutations in amino acid in codon 596 (E596K) and another one in codon 604 (S604F). The S604F mutation was not found in other viral sequences from GenBank. Ten other mutations occurred between codons 377 and 594 in eight samples, including the A594V mutation in a renal transplant patient who ended up dying.It was concluded that the prevalence of antibodies and the epidemiological profile of the group were similar to those observed in populations of developing countries; the viral infection rates are related to viral reactivation, being underestimated by serology; sequence analysis revealed significant genetic diversity in the samples examined; detection of A594V mutation suggests circulating strains with resistance mutation.
Acesso aberto (Open Access)
Correlação dos níveis séricos e dos polimorfismos nos genes de citocinas (TNF-α, INF-γ, TGF-β1 e IL-10) com a apresentação clínica da hepatite B crônica
(Universidade Federal do Pará, 2010) CONDE, Simone Regina Souza da Silva; VALLINOTO, Antonio Carlos Rosário; http://lattes.cnpq.br/3099765198910740
Chronic hepatitis B has a wide spectrum of clinical manifestations resulting from various factors, such as the pattern of secretion and polymorphism in cytokine genes. This work aims to correlate the TNF-α -308G/A, INF-γ +874A/T, TGF-β1 -509C/T e IL-10 -1081A/G polymorphisms and serum levels of these cytokines with the clinical presentation of hepatitis B. It was selected 53 consecutive cases of hepatitis B divided into group A (inactive carrier= 30) and B (chronic hepatitis / cirrhosis= 23). As a control, we selected 100 individuals anti-HBc and anti-HBs positives. Serum levels of cytokines were determined by enzyme immunoassays (eBiosceince, Inc. California, San Diego, USA). The gene amplification of cytokines was carried out by PCR and histopathological analysis followed by METAVIR classification. It was identified that genotype TNF-α -308GA was more prevalent among B group than controls and that presence of A allele increased the risk for chronic disease (OR= 2,6). The serum levels of INF-γ and IL-10 were higher (p <0,001) in controls than others groups A and B and the TGF-β1 levels were lower (p < 0,01) in controls. It was noted that inflammatory activity > 2 correlated with higher levels of TNF-α and IFN-γ (p<0.01) and fibrosis > 2 with higher levels of INF-γ (p <0.01). In the studied population, lower INF-γ and IL-10 levels and higher TGF-β1 level were associated with chronic hepatitis B, and that the presence TNF-α -308 A allele increased in 2,6 the risk for chronic disease .