Programa de Pós-graduação em Medicina Tropical e Saúde Pública

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Toxoplasmose gondii: soroprevalência, isolamento e virulência de cepas obtidas de galinhas caipiras (Gallus domesticus) comercializadas em feiras livres do município de Goiânia
(Universidade Federal de Goiás, 2007-02-27) Aleixo, Eduardo da Costa Alves; Castro, Ana Maria de; http://lattes.cnpq.br/9232309971000621; Soares, Joanna Darc Aparecida Herzog; http://lattes.cnpq.br/6933335902851476; Soares, Joanna Darc Aparecida Herzog; Gomes, Abraão Garcia; Bezerra, José Clecildo Barreto
In this study, 50 free-range chickens were obtained from free fairs of the city of Goiânia, its serum were tested for antibodies anti T. gondii with the modified agglutination test (MAT), its heart and brain had been processed and inoculated in groups of 6 mice. Samples of the organs of the birds and the surviving mice had been sectioned and stained with Hematoxylin-Eosin for the search of Toxoplasma-like cysts structures. The serum of the surviving mice was analyzed by the indirect immunofluorescence reaction (RIFI). The isolated ones had been submitted to the study of virulence for mice by inoculations with different concentrations of tachyzoites. 25 birds (50%) had revealed positive (MAT ≥ 1:5) getting a total of three isolated. Toxoplasma-like cysts structures had been found in histologic cuts of eight birds (16%) and organs of mice of two groups (2%). Of the 25 experiments with inoculeted of positives birds, in 11 (44%) it had the detention of IgG anti-Toxoplasma. In the samples of negatives birds, evidences of the presence of Toxoplasma had not been found. Of the three isolated, two had been lethal for mice from concentrations of a thousand tachyzoites., while another one was 100% fatal one from the concentration of ten a thousand tachyzoites.
Avaliação da ocorrência de rotavírus do grupo a após a implantação da vacina oral de rotavírus humano - VORH e análise comparativa das amostras circulantes antes e após a implantação da VORH em Goiânia – Goiás
(Universidade Federal de Goiás, 2011-04-16) Almeida , Tâmera Nunes Vieira; Fiaccadori , Fabíola Souza; http://lattes.cnpq.br/0993842409303174; Cardoso , Divina das Dôres de Paula; http://lattes.cnpq.br/9770835116155857; Cardoso , Divina das Dôres de Paula; Ulhoa , Cirano José; Pereira , Maristela
The group A rotaviruses (RVA) are recognized as the main viral agents of acute childhood gastroenteritis, and due to the high morbidity-mortality rates vaccination has being considered the best alternative for prevention and control of RVA. In Brazil, in March, 2006 the Ministry of Health included the human rotavirus oral vaccine – VORH, which was developed from a monovalent attenuated strain G1P[8], in the National Immunization Program. In this context, the present study aimed at the investigation of the occurrence of RVA infections in the city of Goiânia after the implementation of the VORH, as well as the comparative analysis of the RVA circulating strains during the pre- and post-vaccination periods. For the RVA identification, 65 fecal samples obtained from children with acute gastroenteritis, in the period from 2008 to 2009, were tested by an immunoenzymatic assay and by polyacrilamide gel electrophoresis, with a total detection rate of 16.9% (11/65). After molecular characterization, the G2 genotype was identified in 10 samples, and four of those were considered as G2P[4] genotype. For the comparative analysis, the G2P[4] samples, as well as other 15 samples, obtained in the pre- and post-vaccination periods, were submitted to genomic sequencing of the coding regions for the proteins VP6, VP7 and NSP4. The molecular characterization of the VP7 gene showed that the G1 samples belonged to lineages I and II, sublineages d and b, respectively, and that all the G2 samples belonged to lineage II, with the differentiation of three sublineages, a, c and d, which were correlated with the collection periods. Regarding the VP6 genogroups and the NSP4 genotypes, a predominance of genogroup I and genotype A in postvaccination period was observed, whereas a predominance of genogroup II and genotype B was identified in the period before de vaccine implementation. The association between the G and P genotypes with VP6 genogroups and NSP4 genotypes revealed the predominance of the G1-P[8]-II-B combination in the pre-vaccination period, and the association G2-P[4]-I-A in the post-vaccination period, which suggests the substitution of these combinations after the implementation of the VORH.
Reconhecimento dos Antígenos Recombinantes MPT-51 e GlcB do Mycobacterium tuberculosis por Anticorpos Séricos de Indivíduos com Tuberculose Ativa
(Universidade Federal de Goiás, 2007-02-23) ALMEIDA, Cristina de Melo Cardoso; KIPNIS, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987
Tuberculosis (TB) caused by the Mycobacterium tuberculosis is responsible for more than 2 million deaths annually in the world. Although one third of the world population carries the bacilli, only 5% of the infected people develop active disease. 80% of TB cases are concentrated among 21 countries and Brazil is one of them. Due to the flaws existent inherent to the TB diagnostic, many works try to discover TB antigens to be used on an ELISA assay to detect active TB in underdeveloped countries, endemic for other mycobacterias such as leprosy (L). Several researches are identifying bacilli proteins obtained under nutritional culture stress, in order to mimic the intracellular milieu. It has been selected the immunodominant ones as a disease marker. The objective of this work was to characterize the humoral immune response among TB patients to rMPT-51 and rGlcB using an ELISA indirect test. It was included voluntaries that were HIV negative and the ones without pregnancy, chronic disease or under immunosuppressant treatment. Serum IgM and IgG against MPT-51 and GlcB recombinant antigen from 49 patients with active tuberculosis were measured by indirect ELISA, paired by sex and age with: healthy PPD negative individuals (controls) and lepromatous leprosy patients (LL). Patients with TB (0.810±0.319) showed higher levels of IgM against rMPT-51 than both LL individuals (0.454±0.195) and control (0.448±0.162) with statistical significant, p= 0.001 and p<0.001) respectively. These test showed 96.9% specificity and 67.3% sensitivity. Conversely, tuberculosis, controls, and LL individuals showed lower of MPT-51 IgG levels, which could not be distinguished among the groups. rGlcB antigen was able to distinguished TB patients from controls for IgM levels (specificity, 95.9 % and sensitivity, 8.2% ) and IgG levels (specificity, 99% and sensitivity, 18.2%). In order to evaluate the profiles of IgM and IgG against rMPT-51 and rGlcB before and after chemotherapy, the sera from 11 patients was collected and paired according to the treatment status (before or after). IgM and IgG against MPT-51 remained with the same profile levels before and after the treatment. The levels of serum IgG against rGlcB clearly diminished after the chemotherapy (p<0.01). Our results suggest that serum IgM levels against recombinant MPT-51 could to be useful to distinguish between active TB, controls and LL individuals. In addition, after TB treatment the IgG response to rGlcB diminished suggesting that it could be used to follow up of the TB treatment
Papel de citocinas e da leptina na imunopatologia da fase aguda na infecção experimental pelo trypanosoma cruzi em camundongos nocautes para IL-4 e IFN-γ
(Universidade Federal de Goiás, 2017-04-07) Almeida, Vera Lucia Lima de; Lino Júnior, Ruy de Souza; Celes, Mara Rúbia Nunes; http://lattes.cnpq.br/1234213665964187; Machado, Juliana Reis; http://lattes.cnpq.br/5289363102869037; Machado, Juliana Reis; Oliveira, Flávia Aparecida de; Castro, Ana Maria de
Chagas disease is a parasitic disease caused by the Protozoan Trypanosoma cruzi. Approximately 30% of infected individuals develop cardiac manifestations. Among the factors involved in this evolution highlights the role of inflammation. Evidence points to an important relationship of adipokines with the cellular immune response and inflammation. The present study is objective is to evaluate the role of cytokines and leptin on immunopathology of acute phase of experimental infection by T. cruzi. For this we used C57Bl/6 WT mice, C57Bl/6 WT KO IFN-γ, Balb-c WT and Balb-c KO IL-4 100 infected with of Colombian strain forms of T. cruzi and non-infected controls was analyzed in the hearts the presence of T. cruzi, fibrosis, inflammatory infiltrate, production of cytokines in situ and serum levels of leptin, TNF-α and IL-10. Noted greater tissue parasitism in C57Bl/6 animals KO IFN-γ, but no difference in the intensity of the inflammatory infiltrate and percentage of fibrosis. Regarding cytokines C57Bl/6 mice IFN-γ KO infected showed increased TNF-α in situ, as well as the Balbc KO IL-4. Despite greater expression in situ, systemically animals Balb-c KO IL-4 have shown to be good producers of TNF-α and IL-10 as well as the C57Bl/6 KO IFNγ. However, these last have shown to be good producers of serum leptin, as well as the Balb-c. Thus, our results indicate that in the absence of IFN-γ mice infected can' not control the tissue parasitism. However, feature increased in situ of TNF-α and serum leptin as a compensatory mechanism of the Th1 response deficiency.
Valor do teste de avidez da IgG como marcador de doença aguda ou crônica e de transmissão vertical na toxoplasmose
(Universidade Federal de Goiás, 2009-06-26) Alvarenga, Fernanda Rassi; Avelino, Mariza Martins; http://lattes.cnpq.br/7890944202988947
Toxoplasmosis is a parasitic disease widespread around the world caused by Toxoplasma gondii. Infection acquired during pregnancy may cause intrauterine damage and sequelae in the newborn. Serological testing for IgG/IgM anti-Toxoplasma antibodies may fail to differentiate between recent and past infection. Despite that, rapid diagnosis of acute infection during pregnancy allows rapid treatment and prevents or attenuates congenital toxoplasmosis. PURPOSE: to establish the frequency of acute toxoplasmosis in pregnant women, the vertical transmission rate and the value of specific IgG-avidity test to date infection in pregnancy; to evaluate the relationship between IgG-avidity and congenital toxoplasmosis. MATERIAL AND METHODS: this report summarizes a retrospective study performed on 235,993 pregnant women attended by “The Pregnancy Protection Program” – public health system (SUS) of the State of Goiás – Brazil, from January 2004 to December 2007. ELISA (IgG / IgM) and IgG-avidity test were performed for maternal screening of toxoplasmosis. Fetal and newborn investigation of the infection was performed by “The Toxoplasmosis Vertical Transmission Control Program” protocols. The association between data was statistically analyzed by the x2 test (p < 0,05 was considered statistically significant). RESULTS: the frequency of IgM-positive among pregnant women studied population was 0,7%. Among IgM-positive women, only 207 (12,5%) performed the screening test in first 3-month period of pregnancy and 91% of pregnant women presented high avidity ( > 40%). The vertical transmission rate was 62%. There was no statistically significant relationship between higher (> 40%) or lower (≤ 25%) IgG-avidity test and presence of congenital infection (p= 0,08 e p= 0,57, respectively). There was no statistically significant association between maternal diagnosis in first trimester, low avidity and vertical transmission rate. CONCLUSIONS: the frequency of IgM-positive in pregnant women was lower than Brazilian rates founded in other studies. The study showed high persistent vertical transmission rate besides prenatal management and treatment. The IgG-avidity was not useful to predict vertical transmission. These results indicate that the IgG-avidity test must be not carried out in all IgM-positive pregnant women in the State of Goiás-Brazil, as a confirmatory test for the diagnosis of maternal toxoplasmosis.
Perfil epidemiológico e clínico das meningites infecciosas em idosos, no Hospital de Doenças Tropicais – HDT/AA Goiânia, Goiás, Brasil, 2000 - 2006
(Universidade Federal de Goiás, 2008-08-25) Alvarenga, José Alberto; Almeida Neto, Joaquim Caetano de; http://lattes.cnpq.br/3444498706763045
It is estimated that until the year 2020, in Brazil, 13 % of the population will be constituted by the elderly (population who are above 60 years old), counting 30 million people. The frequency and the gravity of the infectious diseases spread with age. Among these days, infectious diseases are the third cause of death after the sixty’s. The physical changes and the lack of immunological response in the elderly worsen day after day, favoring the infections’ gravity. Moreover, chronic diseases treated with immunosuppressor drugs and other, repeated hospitalizations and nutritional deficiency are addictive factors which explain this kind of diseases to grow worse in this specific population. This dissertation is about a retrospective essay with a descriptive analysis raising medical documents from patients with infectious meningitis in the Hospital de Doenças Tropicais (HDT), State of Goiás, Brazil, and aimed to evaluate the epidemiologic and clinical profile of the infectious meningitis in the elderly, the level of clinical suspicion and confirmation in health services in our State. About 110 cases of meningitis in elderly were notified between the years 2000 and 2006. From all cases, 64 (58,71%) were confirmed, 64,0% of non-specified infection meningitis, 14,06%, fungus meningitis (4,68%), pneumococcol meningitis (12,5%), tuberculosis meningitis (3,12%) and meningococcal meningitis(1,5%). The mortality was higher in non-specified meningitis (65,21%), and fungus, tuberculosis and pneumococcol meningitis presented the highest lethality. The long period between the clinical suspicion of meningitis, with 6,41 days until the diagnose was confirmed, delayed treatment and surely contributed to raise the number of the infections and deaths. The unsuitable medical graduation in elderly’s support has delayed diagnose, extending the interval between the beginning of the symptoms and the effective treatment, and so on, increasing the mortality index of this population. The proposal of this essay is to build a preferable attendance service for the elderly, with basic knowledge in infectious diseases in this particular population, favoring the fast diagnose and the effective treatment in order to reduce the mortality in this age group.
Avaliação clínica, laboratorial e histopatológica do efeito de drogas imunossupressoras na reativação da toxoplasmose crônica em modelo murino com a cepa ME 49 no camundongo BALB/c
(Universidade Federal de Goiás, 2008-03-28) Alves, Fabiana Santiago Aleixo; Lino Junior, Ruy de Souza; http://lattes.cnpq.br/0372118837748010; Soares, Joanna Darc Aparecida Herzog; http://lattes.cnpq.br/6933335902851476; Garciazapata, Marco Tulio Antonio; http://lattes.cnpq.br/3672512339058369; Garciazapata, Marco Tulio Antonio; Lacerda, Elisângela de Paula Silveira; Carvalhaes, Mara Silva
This work was evaluated for the potential reactivation of Toxoplasmosis in murine model similar to human immunosuppression being developed in BALB/c mice, which were 40 days infected with 20 cysts of ME 49 strain of Toxoplasma gondii and after 60 days was initiated treatment with immunosuppressant drugs, Azathioprine in dosage of 10mg/kg five times a week in days, Dexametasone in dosage 2.5mg/kg per day per mouse, three times per week on alternate days, Cortisone acetate in 50mg twice a week in subcutaneous injection of cortisone acetate alone, or associated with its proper controls. Treatment was continued for 28 days. The use of the Dexametasona or Azathioprine isolated or associated with, a factor not caused by reactivation of serological tests, clinical or histopathological but associated with cortisone acetate led to a clinical diagnostic framework of a voluntary recall because of injuries in epidermal 62.5% of the mice that lot compared to its proper controls, however mortality was not observed in any of the groups tested
Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
(Universidade Federal de Goiás, 2014-04-30) Alves, José Vitor Ferreira; Dorta, Miriam Leandro; http://lattes.cnpq.br/3933395097851681; Dorta, Miriam Leandro; Moraes, Sandra do Lago; Dias, Fátima Ribeiro
Leishmaniasis is a group of diseases with distinct clinical, histopathological and immunological characteristics, caused by parasites belonging to the Leishmania genus. The serological tests used until the moment have several limitations. There are a great interest to identify immunogenic proteins of Leishmania to be tested as potential antigens for the development of techniques for the diagnosis of American cutaneous leishmaniasis (ACL). The aim of this study was to produce and purify the recombinant proteins "Leishmania activated C kinase" (rLACK); "Thiol Specific Antioxidant" (TSA), "Leishmania elongation initiation factor" (LeIF) and "Leishmania braziliensis stress inducible protein 1" (LbSTI) to evaluate the antigenicity. The recombinant proteins were produced by recombinant DNA techniques as described by Salay et al. (2007). To perform the ELISA, L.(V.) braziliensis (MHOM/BR/1975/M2903) and L.(L.) amazonensis (IFLA/BR/67/PH8) species were cultivated and the antigenic extracts and recombinant proteins were used as antigens. Sixty serum samples from patients with ATL assisted at Anuar Auad hospital, Goiânia, Goiás, were assayed, and from them, 45 were from patients with localized cutaneous leishmaniasis (LCL) and 15 were from mucosal leishmaniasis (ML). To analyze the specificity of the response of total extract and the recombinant proteins, sera from patients with other pathologies were tested. The total extract of L. (L.) amazonensis, rLACK, rTSA, rLbSTI and rLeIF showed a sensitivity of 85%, 75%, 70%, 76.7% and 56.1% respectively. The specificity of total extract of L. (L.) amazonensis, rLACK, rTSA, rLbSTI, rLeIF were 72.5%, 80%, 60%, 30% and 62.5% respectively. Thus, these results showed that recombinants proteins and total extract of L. (L.) amazonensis were antigenic and the total extract of L. (L.) amazonensis and rLACK were the antigen that had the best sensibility and specificity.
Análise das Bases Moleculares da Resistência à Isoniazida e Rifampicina em Cepas Obtidas de Pacientes com Tuberculose no Estado de Goiás
(Universidade Federal de Goiás, 2010-03-11) ALVES, Sueli Lemes de ávila; KIPNIS, André; http://lattes.cnpq.br/4434965360286741
Multidrug-resistant tuberculosis is a challenge worldwide. Rapid diagnosis by molecular techniques can provide a more aggressive and appropriate initial therapy. This study aimed to analyze the molecular basis of resistance to isoniazid (INH) and rifampin (R) of Mycobacterium tuberculosis strains isolated from cases of human tuberculosis in Goiás and to genetically determine the causes of the observed resistances. Of the 4.607 cultures for mycobacteria processed in the period of September of 2005 and December of 2007, 24 isolates from 16 patients resistant to at least H and/or R were analyzed. We compared the results obtained by phenotypic tests with mutations in key genes responsible for the development of resistance to these drugs, the rpoB gene for isolates resistant to R and katG gene for strains resistant to H. Seventy one percent of the isolates were resistant to H, and the mutations involved with resistance observed in the katG gene were in codon 315 (41%). The most frequent mutations observed in the rpoB gene of the R resistant isolates (71%) were in codons 456 (76.5%) and 451 (17.6%). Our findings are similar to those reported in the literature. We conclude that the percentage of agreement between genotypic and phenotypic tests was 41% for H and 94% for R considering the number of isolates and 40% and 91%, respectively considering the number of patients.
Avaliação do processo de descontaminação de brocas odontológicas e seu impacto no controle de infecção
(Universidade Federal de Goiás, 2006-03-31) ANDERS, Patrícia Staciarini; TIPPLE, Anaclara Ferreira Veiga; http://lattes.cnpq.br/4288704233343920; PIMENTA, Fabiana Cristina; http://lattes.cnpq.br/2230554075502158
Objective: to describe the process of decontamination of burs to assess the microbial contamination of burs after-processing that were available to the use to isolate and to identify the possible microorganisms Methods: This study was conducted in 110 private dental offices of the central area of Goiânia-Goiás during the period of March/2004 to August/2005 using a check-list measure of dry heat sterilizer temperature and microbial burs tests The burs were seeded in brain heart infusion broth incubated at 37ºC for 20 days and subcultured on specific agar to isolate microorganisms The isolates were identified by micro/macroscopic characteristics subcultured on specific agar biochemical/enzymatic test and automation technique (MicroScan ®) Results:A total of 110 burs were evaluated and 35 (31.8%) were contaminated Fungi were detected in 13 (30.2%) burs [Aspergillus sp (27.9%) and Micelia (2.3%)]; Gram-positive cocci (staphylococci) represented 13 (30.2%) isolates [2.3% Staphylococcus aureus and 27.9% (12) coagulase negative staphylococci] nine (20.9%) isolates were Gram-positive bacilli and eight (18,6%) fastidioso microorganisms Considering the obtained results some factors detected could be interfered in the burs sterilization ineficiency: enzymatic detergent inadequate use abrasive products use inadequate dry heat sterilization time and temperature multiple use burs kits and interruption the asseptical chain after sterilization Conclusion: The frequency of contaminated burs was high (31.8%) and it was detected failures in operational steps of burs processing and/or after sterilization
Prevalência da infecção pelo vírus da hepatite C e práticas de risco em homens que fazem sexo com homens em Goiânia-Goiás, empregando o método respondent-driven sampling (RDS)
(Universidade Federal de Goiás, 2016-02-29) Andrade, Andreia Alves de; Martins, Regina Maria Bringel; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787399H4; Martins, Regina Maria Bringel; Carneiro, Megmar Aparecida dos Santos; Minamisava, Ruth
Hepatitis C is the most common cause of chronic liver disease. Men who have sex with men (MSM) may be at increased risk of acquiring infections transmitted by parenteral and sexual routes, such as hepatitis C. Despite the relevance of this subject, only one investigation regarding hepatitis C virus (HCV) infection in MSM was conducted in Brazil. Therefore, this study aimed to investigate the prevalence of HCV infection and risk practices among men who have sex with men in the City of Goiânia, Goiás. Cross-sectional study conducted in 522 MSM in Goiania, recruited by respondent-method driven sampling (RDS), from March to November 2014. After signing the informed consent, participants were interviewed about sociodemographic and risk behaviors/practices for HCV infection and then blood samples collected. All samples were subjected to determine the levels of alanine aminotransferase and aspartate aminotransferase (ALT and AST) and anti-HCV marker detection. Samples that had high levels of ALT/AST and/or were anti-HCV positive were tested for HCV RNA by reverse transcription-polymerase chain reaction (RT-PCR), and the positive samples were genotyped by a line probe assay (LiPA). Of 522 samples, four were anti-HCV positive and 14 had high levels of ALT/AST. Of these, only two (Y-421/anti-HCV positive and Y-180/anti-HCV negative/elevated ALT and AST) were HCV RNA positive, and then were genotyped by LiPA as genotype 1, subtypes 1a and 1b. Thus, five samples were anti-HCV and/or HCV RNA positive, resulting in an adjusted prevalence of 1.71% (95% CI: 0.32-8.55) for HCV infection among MSM in Goiânia-GO. Despite it is a low prevalence, many risk behaviors/practices were reported by the studied individuals (tattoo/piercing, sharing of personal hygiene tools, illicit drug use, multiple sexual partners in lifetime, sex with more than one partner in the same relationship, sex with a drug user partner, non-use or occasional condom use during anal sex, alcohol/drug use during sex, among others), which may characterize the target population as potentially vulnerable to infectious diseases transmitted by parenteral and sexual routes.
Análise da variação de IgG3 e IgG total específicas para antígenos de Leishmania (Viannia) braziliensis em soro de pacientes com leishmaniose cutânea antes e após o tratamento
(Universidade Federal de Goiás, 2015-09-18) Andrade, Douglas Oliveira; Oliveira, Milton Adriano Pelli de; http://lattes.cnpq.br/2152513705182408; Oliveira, Milton Adriano Pelli de; Pereira, Ledice Inácia de Araújo; Lino Júnior, Ruy de Souza
Leishmaniasis are a group of endemic diseases in 98 countries, with an estimated incidence up to 1.6 million cases per year (0.7-1.2 million of cutaneous leishmaniasis and 0.2 to 0.4 million visceral leishmaniasis). The criteria for the cure of american tegumentary leishmaniasis adopted nowadays is the complete healing of the lesion. IgG3 subclass anti-leishmania antibodies decrease faster than the total IgG antibodies in cured patients. This study was aimed to identify differences in the recognition pattern of L. (V.) braziliensis protein by total IgG or IgG3 antibodies obtained from cutaneous leishmaniasis infected patients’ serum before and after treatment. Sera of 15 patients infected with L. braziliensis attended at the Tropical Diseases Hospital Anuar Auadi were tested by ELISA against crude extracts of L. braziliensis. When all patients were analyzed, it was observed that anti-leishmania IgG3 absorbance levels measured by ELISA were similar to the controls levels at 6 month. The absorbance levels of total antibodies in control and infected patients reached similarity only at 12/18 months. After split patients into group A: healing of the lesion before one month and B: healing of the lesion after one month, it was observed that absorbance levels in ELISA for IgG3 in group A was similar to the controls levels at the time of diagnosis. It was also observed, by Western Blotting, that the IgG3 reactivity of IgG3 to 49 kDa protein waspresent mainly for group B patients. The 28 kDa protein showed IgG3 reactivity mainly in group B the healing os lesions and the 14 kDa protein showed IgG3 reactivity in some controls our data suggest that the use of 49 kDa protein in ELISA may be useful for diagnosis and following up of patients and the 14kDa protein may be important in false-positive results.
Expressão heteróloga da protease Rv 2467 de Mycobacterium tuberculosis
(Universidade Federal de Goiás, 2018-06-12) Andrade, Rayanny Gomes de; Kipnis, Ana Paula Junqueira; http://lattes.cnpq.br/1252262903952987; Kipnis, André; http://lattes.cnpq.br/4434965360286741; Kipnis, André; André , Maria Cláudia Dantas Porfírio Borges; Steindorff, Andrei Stecca
Mycobacterium tuberculosis is the etiological agent of tuberculosis (TB), which is responsible in 2015 for 10.4 million new cases and 1.8 million casualties registered worldwide. To be able to avoid the defense barriers of the host, the bacteria evolved mechanisms to modulate or change the environment it is in, thereby, the secretion of bioactive molecules is an efficient strategy to do so. Among the bioactive molecules produced by M. tuberculosis, proteases have a crucial role to a successful infection, as they are involved in several important biological processes of the bacteria, such as DNA replication, cell proliferation and antigen processing. Thus, proteases are good targets for the development of new anti-TB drugs or as possible vaccine antigens. This work aimed to clone and express the gene Rv2467, a putative protease with aminopeptidase activity from M. tuberculosis. To achieve this goal, oligonucleotides design were made to amplify the gene Rv2467 by Polimerase Chain Reaction (PCR), which was cloned in the pGEM-T easy vector. The clone Rv2467 gene was then transfere to the expression vector pET-28a. The recombinant protein was expressed from the recombinant plasmid pET-28a/Rv2467 in Escherichia coli BL21 (DE3) pLysS through induction with IPTG. The recombinant protein, with the expected size of 94KDa, was expressed and subjected to the purification process by nickel affinity chromatography. The recombinant protein was partially purified only in its denatured form. The low yield of purified recombinant protein raised the possibility of histidine tail loss. To verify that, Western Blotting with anti-histidine antibody was made and it was verified that the antibody did not recognize the target Rv2467 protein, which made it impossible to acquire the pure protein, not allowing further characterization tests. Additionally, a literature review was performed about Mycobacterium tuberculosis proteases that are involved in virulence mechanisms to make a manuscript to be submitted to a scientific journal
Reposicionamento in silico de novos fármacos contra o schistosoma mansoni
(Universidade Federal de Goiás, 2016-03-14) Arantes, Morgana Elias; Andrade, Carolina Horta; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4745602P1; Bezerra, José Clecildo Barreto; http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781675U6; Andrade, Carolina Horta de; Cravo, Pedro Vitor Lemos; Braga, Rodolpho Campos
Schistosomiasis is a neglected tropical disease caused by parasites of the genus Schistosoma. In Brazil only Schistosoma mansoni transmits this disease. The World Health Organization estimated in 2012 approximately 249 million people at risk of acquiring this disease around the world. The main strategy to control this disease is the treatment of individuals living in endemic areas with Praziquantel. The drug praziquantel is used on a large scale in the treatment of schistosomiasis and currently there are reported cases of resistance, indicating the need to discover new drugs. In silico drug repositioning is a strategy that reduces the time and cost in the search of anti-schistosomal agents. This work used bioinformatics tools and methodology based on previous studies, as a means to identify potential new S. mansoni targets and drug homologs, consequently identifying potential new schistosomal drugs. A list was compiled with S. mansoni potential targets that are part of essential processes in the database TDR and the targets that are part of the tegument were obtained in the scientific literature. The file with S. mansoni targets contained 1376 targets, and of these only 61 targets associated with 399 drugs had homology with drug targets. After removal of duplicate drugs, drugs found in previous studies and after the analysis of the conservation of the active site only 28 S. mansoni targets associated with 102 drugs had 60% or more of the active site conserved. Some of the drugs had activity and are interesting to validate this study such as: artemether, lumefantrine, meloxicam. Among the drugs found 18 drugs were selected to be tested using the following criteria: low toxicity in vivo, expired patent and a value of log P interesting for oral administration.
Testes de microdiluição em caldo e diluição em ágar para avaliação da suscetibilidade in vitro de dermatófitos
(Universidade Federal de Goiás, 2007-05-31) ARAUJO, Crystiane Rodrigues de; SILVA, Maria do Rosário Rodrigues; http://lattes.cnpq.br/7119226630434725
Dermatophytes are keratinophilic fungi that colonize and invade the stratum corneum of the skin, hair and nails causing the dermatophytosis. An increasing number of antifungal agents has become available for the treatment of dermatophytosis, however not all species have the same susceptibility pattern and may occur relative or absolute resistance of some dermatophytes. The document M38-A developed by Clinical and Laboratory Standards Institute (CLSI) for determining the minimal inhibitory concentration (MIC) of different antifungal agents against filamentous fungi, has not included the dermatophytes. The broth microdilution method has been evaluated by various researchers, and some parameters as inoculum size, temperature and duration of incubation and endpoint determination has been investigated. In this study, the in vitro activity of fluconazole, itraconazole, ketoconazole, griseofulvin and terbinafine against 60 dermatophyte isolates, belong to three species, using the broth microdilution technique, with modifications at temperature and incubation time was used. Additionally, the MIC values obtained by broth microdilution method were compared with those obtained by the agar dilution technique. The results obtained by broth microdilution method showed that all isolates produced clearly detectable growth at 28oC and the MIC values could be determined after 4 days of incubation for the isolates of Trichophyton mentagrophytes and 5 days for T. rubrum and Microsporum canis isolates. Itraconazole, ketoconazole and terbinafine had the lowest MIC values (0.03 μg/ml) for 33.3%, 31.6% and 15% of the isolates, respectively. A good concordance was observed between the agar dilution and broth microdilution methods. The levels of agreement were 91.6% with ketoconazole and griseofulvin, 83.3% with itraconazole, 81.6% with terbinafine and 73.3% with fluconazole for all the tested isolates. In summary, the results of this study suggest that an incubation time of 5 days and temperature at 28oC used in broth microdilution and agar dilution methods can contribute to define and to better interpret the MIC values. Beside, until a reference method for testing the susceptibilities of dermatophytes is standardized, the similar results with broth microdilution method become the agar dilution useful for testing the susceptibility of these fungi.
Investigação epidemiológica e molecular da infecção pelo vírus da hepatite C em assentados nos estados de Goiás e Mato Grosso do Sul, Brasil
(Universidade Federal de Goiás, 2014-02-25) Araújo, Lyriane Apolinário de; Carneiro, Megmar Aparecida dos Santos; http://lattes.cnpq.br/8398563469665169; Carneiro, Megmar Aparecida dos Santos; Martins, Regina Maria Bringel; Aires, Rodrigo Sebba
The hepatitis C virus (HCV) infection is considered one of the main causes of chronic liver disease, being responsible for great disease burden to the Health Systems Worldwide. According to the World Health Organization, there are approximately 150 million people chronically infected by the HCV in the world. There are only a few studies that have evaluated the HCV occurrence in people that live in settlements, and most of these families are composed by a large number of individuals that have little formal education, that live under poor environmental and sanitation conditions, beyond also usually have no access to adequate health care, due primarily to the difficulties in reaching the health service systems, but also because of the poor quality of the health services. Therefore, the aim of this study was to estimate the prevalence of HCV infection, to analyze the characteristics of the population associated with the infection and to identify the HCV genotypes/subtypes in individuals that live in settlements in the States of Goiás and Mato Grosso do Sul, Central Brazil. The population was consittued by 921 individuals. They were all interviewed and one blood (serum) sample from each person was collected. All sera samples were screened for anti-HCV using an Enzyme Linked Immunosorbent Assay (ELISA). All ELISA positive samples were submitted to RNA extraction and tested by Polymerase Chain Reaction (PCR). All PCR positive samples were further genotyped using the Line Probe Assay (LiPA). The mean age of the population was 39 years old, composed mainly by Caucasian married women, with a low level of formal education, and low family income. Of the 921 samples, four were anti-HCV positive, resulting in an overall prevalence of 0.43% (CI 95%: 0.14-1.19). The viral RNA was detected in all anti-HCV positive samples and the genotyping revealed the presence of the genotype 1, subtype 1a. The HCV infected individuals reported as risk caracteristics for HCV infection: sharing sharp personal hygiene objects, history of surgical intervention, blood transfusions before to 1993, sex without protection, history of sexually transmitted disease and imprisonment. This study showed a low prevalence of HCV infection in this population. This study showed a low prevalence of HCV infection in this population. However, epidemiological investigations are important to verify the circulation of the virus , especially in settled populations, which are often on the margins of public health services.
Prevalência de infecção pelos vírus dengue em parturientes e neonatos, Goiânia-Goiás, 2009-2010
(Universidade Federal de Goiás, 2010-12-21) Argolo, Angela Ferreira Lopes de Teive e; Féres, Valéria Christina Rezende; Martelli, Celina Maria Turchi; http://lattes.cnpq.br/5867052489026059; Braga, Maria Cynthia; Siqueira Júnior, João Bosco; Martelli, Celina Maria Turchi
The research about the epidemiology and the pathogeneses of dengue virus during the gestational period is essential to the understanding of clinical dengue severity among women and their offspring. It has been postulated that children born from women with antidengue antibodies may develop severe dengue during the first viral dengue exposure. The objectives were (a) to determine the prevalence and incidence of dengue infection in pregnant women and their neonates in two public hospitals in the city of Goiania, Goias State, Central Brazil, (b) to assess the percent of newborns with maternal IgG and DENV vertical transmission, (c) to evaluate the association of maternal recent dengue infection (IgM) with the characteristics of the delivery and neonates. A cross sectional study was conducted to assess recent (IgM) and previous (IgG) markers of dengue infection among pregnant women and newborns. The survey was conducted in two public hospitals in the city of Goiania (~1.2 million inhabitants), Central Brazil, from December 2009 to May 2010. We recruited women after giving birth. Women were interviewed to collect data about previous dengue infection, symptoms of dengue during gestational period, pregnancy period, delivery mode and outcomes pregnancy. Blood samples were collected from women and neonates (umbilical cord). Previous infection was defined by the positivity to IgG serology (PAN BIO) and recent infection by IgM (PAN BIO). The detection of NS1Ag and viral nucleic acid (RT-PCR) were performed in the subsample of participants who reported specific symptoms of dengue fever ten days previous to the delivery time. Prevalence and incidence estimates were calculated with 95% Confidence Intervals (95% CI). The agreement between the positive and negative serological results of the pair of women and their concepts was calculated (Kappa Index). Positive and Negative Predictive values between the history of previous dengue and the results of serological markers were calculated. A case-control analysis was performed to assess risk factors among IgM positive women (cases) and the outcomes of pregnancy compared to the seronegative women (controls). Analysis was performed using SPSS 17.0 e o Epi Info 6.04. The investigation was approved by the regional Ethical Committee and all participants signed informed consent according to the Brazilian regulation. A total of 505 women and 505 neonates were investigated. The mean age of women was 25.8 (SD = 6.4), 83.6% of ABSTRACT xv deliveries were between 37 and 41 weeks of pregnancy. The prevalence of IgG dengue antibody was 53.9% (95% IC 49.3% - 58.0%) among women. Maternal antibodies were also found in 99.2% of the offspring (Kappa index = 0.96). NS1 antigen was negative among women and one case of DENV-2 was detected. The incidence of recent dengue infection (IgM positivity and RT-PCR) was 8.9% (95% CI 6.6% - 11.8%) among women and 1.6% (95% IC 0.7%-3.2%) among children. Eight neonates had IgM antibodies with evidence of vertical DENV transmission. The positive predictive value of the previous history of dengue related to seropositivity (IgG) was 88%. The outcomes of pregnancy were similar among seropositive and seronegative women in the case-control analysis. In our setting, half of the pregnant women had dengue antibodies. There was high agreement rates between the serological markers detected among the pair of women and their concepts. Acute dengue infection among pregnant women was not associated with the outcomes unfavorable of pregnancies.
Isolamento, identificação e suscetibilidade in vitro de fungos causadores de onicomicose
(Universidade Federal de Goiás, 2010-02-24) ATAÍDES, Fábio Silvestre; SILVA, Maria do Rosário Rodrigues; http://lattes.cnpq.br/7119226630434725
Onychomycosis is the nail infection caused by a wide spectrum of fungi species, including yeasts, dermatophyte and filamentous fungi nondermatophytes (FFND). In this work, patients with lesions nails from Dermatology Department in Clinics Hospital of the Federal University of Goiás were studied. These patients were submitted to mycologic tests and the etiologic agents identified were evaluated on in vitro activity for systemic antifungal agents. During the period of March 2008 to March 2009, 114 patients clinically suspected of having onychomycosis were examinated. The nail samples collected were submitted to direct examination with potassium hydroxide 20% and grown in dextrose Sabouraud agar and specific fungal pathogens agar. The antifungal susceptibility test was performed according to the method of broth microdilution, recommended by the Clinical Laboratory Standards Institute (CLSI), through M38-A and M27-A2 documents. The onychomycosis diagnosis was established in 83.3% (95/114) patients, of which most were women between 40 to 59 years old. The isolated fungi were identified as yeasts in 54.6%; dermatophytes in 28.7%; and filamentous fungi nondermatophytes in 16.7%. Among yeasts, Candida parapsilosis was the most common etiologic agent (52.5%); among dermatophytes, Trichophyton rubrum was the most found (74.2), and among the filamentous fungi nondermatophytes, Fusarium spp was the most isolated genus (44.4%). Both distal and lateral subungual lesions were predominant in cases of onychomycosis by all the agents identified, showing that there was no correlation with the clinical presentation and etiology. Although most of the isolates have showed susceptible to several antifungal agents studied, five Candida spp strains were resistant; one to voriconazole, one species to itraconazole and three to amphotericin B. The in vitro susceptibility testing profile for dermatophytes was similar for each antifungal agent analyzed. The minimal inhibitory concentration (MIC) of itraconazole, ketoconazole and griseofulvin for 50% of isolates were lower than 1 μg/mL, and terbinafine was extremely low, with concentration 0.015 μg/mL for 90% of isolates. The MIC for itraconazole in 90% isolates of FFND was 16 g/mL, while for amphotericin B and voriconazole was 8 g/mL. In summary, this study showed a higher frequency of onychomycosis in women. Candida spp and dermatophytes with FFND emergence, especially of the genus Fusarium, were the main fungi involved. Besides, the presence of resistant fungi to some antifungal agents showed the need of using susceptibility tests for these fungi.
Caracterização molecular de isolados de Trypanosoma cruzi obtidos de mulheres durante a fase crônica da doença de Chagas
(Universidade Federal de Goiás, 2008-02-29) Avelar, Juliana Boaventura; CASTRO, Ana Maria de; http://lattes.cnpq.br/9232309971000621
Chagas disease is a zoonotic disease caused by a protozoan plagued called Trypanosoma cruzi, it is a particular problem in Latin America, where it is estimated that 12 to 14 million people are infected. The populations of Trypanosoma cruzi can be divided into two major groups, the Trypanossoma cruzi I (TCI) and Trypanossoma cruzi II (TCII), the last being more heterogeneous and divided into five subgroups TCIIa, TCIIb, TCIIc, TCIId and TCIIe However, little is known of epidemiological and clinical implications of these ratings. The frequency of vertical transmission of Chagas disease can vary from 1.6 to 18.5% in accordance with the region and the methodology used in the studies. The aim of this study was to genetic characterization of 30 isolates of T. cruzi through the blood of patients with Chagas disease in the chronic phase, pregnant and not pregnant. The genetic characterization was performed with three different markers that amplified regions of the 18S and 24S α DNA ribosomal and mini-exon of the gene that allows classify the groups and subgroups of the parasite and the analysis of genetic variability among isolates of T. cruzi was performed the characterization of the variable region of minicicles of kDNA through the technique of Low-Stringency Single Specific Primer-PCR (LSSP-PCR). The 30 patients studied 75% (25/30) were women and 25% (5 / 30) not pregnant. The patients are the four different Brazilian regions being the States of Bahia 53.3% (16/30), Goiás 40.0% (12/30), Paraíba 3.3% (1 / 30) and the Federal District 3, 3% (1 / 30). The analysis of rDNA allowed show the TCII in 100% of the isolated populations in the regions studied, where the subgroup TCIIb/e has been detected in (96.66%) and TCIId in (3.33%) the latter is a sample of the group Control of the State of Bahia. The genetic variability between the 30 samples was 24.08 ± 14.4% share of bands among pairs. There was no difference between the share of bands patients of Bahia and Goiás, where the value of significance was less than 5%. There was no difference between the percentages of bands showed in analyses by age of the patients, gestational age and in the analysis of the groups of pregnant and not pregnant. In reviewing more than one tube of blood per patient coming evidenced profiles of signatures genomic complex and heterogeneous with 62.5% of bands shared less than 77% and only 37.5% had rates higher than 83% of bands sharing among samples. Suggesting so our isolates showed a great genetic variability.
Análise da expressão das proteínas META2, LRR17, STI1 e TSA em isolados de Leishmania (Viannia) braziliensis obtidos de pacientes com lesão cutânea ou mucosa
(Universidade Federal de Goiás, 2012-02-17) Ávila, Lucilla Ribeiro; Dorta, Miriam Leandro; Oliveira, Milton Adriano Pelli de; http://lattes.cnpq.br/2152513705182408; Castro, Ana Maria de; Afonso, Luis Carlo Crocco
Introduction: Leishmania (Viannia) braziliensis is the most common etiologic agent of cutaneous leishmaniasis in Brazil. About 1 to 10% of patients infected with this species of parasite develop the mucosal leishmaniasis, whose lesions are progressive, destructive and are characterized by a strong specific immune response. The mechanisms for the development of mucosal leishmaniasis are poorly known, but it is possible that proteins associated with virulence of the parasite, such as META2 LRR17 and/or related with the induction of a strong immune response, such as STI and TSA, participate in the pathogenesis disease, causing the appearance of metastases in the mucosa. Objective: To evaluate the expression of proteins META2, LRR17, STI1 and TSA in promastigotes and amastigotes of Leishmania (Viannia) braziliensis derived from cutaneous or mucosal lesions. Methods: It was used three isolates from cutaneous lesions of patients (JCJ8c, and RPL5c SMB7c) and three isolates from mucosal lesions (ASL9m, and JBC8m PPS6m). Amastigotes of the isolates were obtained after inoculation of biopsies in mice knockout in interferon gamma and promastigotes in logarithmic or stationary phase obtained in culture in Grace's medium. The kinetics of growth in culture of isolates was performed by counting daily over ten days in the flow cytometer. The expression of proteins of each isolate was assessed by immunoblotting technique and by flow cytometry, the latter being used only to evaluate the expression of proteins whose expression was significant difference between isolates from cutaneous or mucosal lesions. Results: Promastigotes in stationary phase of the isolates SMB7c and JBC8m were the most expressed the protein META2. The greatest expression of this protein in parasites coincided with more severe lesions, since despite of isolated SMB7c being of cutaneous origin, the patient also presented mucosal lesion, and in the isolated JBC8m the patient presented return of lesions. Proteins LRR17 and STI1 were not expressed in significant amount in both promastigotes and amastigotes of different isolates. TSA protein was expressed at higher levels in promastigotes stationary phase and amastigotes of the xi isolates from patients with mucosal lesions, and this difference was also observed in flow cytometric analysis. Conclusion: The results suggest an association of META2 protein, which is related to the virulence of the parasite, with the generation of more severe lesions in the mucosal, since the highest expression of the protein was isolate from patients with mucocutaneous lesions and patients whose treatment was not effective. TSA, which is associated with antigenicity and virulence of the parasite was also expressed in greater amount in isolates from patients with mucosal lesions. The hypothesis is that, despite TSA protein to induce a protective immune response could be conferring protection in the Leishmania after entering the phagolysosomes by being an antioxidant protein. This protection favored the persistence of parasites and later generation of mucosal lesions.

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