HELICOBACTER PYLORI OIPA VIRULENCE GENE AS A MOLECULAR MARKER OF SEVERE GASTROPATHIES

ABSTRACT Background: Helicobacter pylori is an etiologic agent of gastroduodenal diseases. The microorganism, considered a type I carcinogen, affects about 50% of the global population. H. pylori virulence factors are determinant for the clinical outcome of the infection. The outer inflammatory protein A (oipA) gene encodes an outer membrane adhesin and is related to severe gastropathies, such as gastric cancer. Objective: The aim of this study was to evaluate the association of the oipA gene with the severity of gastroduodenal diseases in dyspeptic patients in region Central Brazil. Methods: The polymerase chain reaction (PCR) was used to determine the presence of H. pylori. Samples positives were used for molecular screening of the oipA gene. Gastropathies were categorized as non-severe and severe diseases. Results: Approximately 68% of patients had H. pylori and 36% were infected with H. pylori oipA+ strains. Infection was significantly associated in patients aged over 44 years (P=0.004). However, there was no association between oipA and patients’ age (P=0.89). Approximately 46% of patients infected with oipA+ strains had some severe illness. Gastric adenocarcinoma was the most frequent severe gastropathy. The H. pylori oipA genotype was inversely associated with the severity of gastroduodenal diseases (OR=0.247, 95%CI: 0.0804-0.7149 and P=0.007). Conclusion: The characterization of possible molecular markers will contribute to personalized medicine, impacting the prognosis of patients

Histological and immunohistochemical evaluations of the bone marrow from femur and sternal manubrium of dogs reactive for leishmaniasis by DPP® and ELISA tests

As the bone marrow is one of the most organs affected by canine visceral leishmaniasis (CVL), samples from this are frequently taken for parasitological tests, with occurrence of myelodysplastic changes, with consequent anemia, leukopenia, and thrombocytopenia. Thus, this study aimed to investigate the histological and immunohistochemical changes in the bone marrow of the femur and sternal manubrium of dogs reactive for leishmaniasis by DPP® and ELISA tests. For this, thirteen canines from the epidemiological routine for CVL carried out by the Directorate of Zoonosis Surveillance of Goiânia (DVZ), GO, Brazil, were subjected to anatomopathological examination. 46.2% of bone marrow samples from the femur showed a higher proportion of the red series, and 53.9% of bone marrow of the sternal manubrium evidenced a higher proportion of the red series. Also, there were varied macrophage hyperplasia, hemosiderosis, and megakaryocytic emperipolesis. Amastigote forms of Leishmania spp. in the bone marrow of the femur and sternal manubrium to histopathological and immunohistochemical evaluations were observed, with good agreement them, but without difference in the parasite intensity between the bone marrow of these anatomical sites. It was concluded that bone marrow of the femur and sternal manubrium of dogs reactive for leishmaniasis by DPP® and ELISA tests has histological changes resulting from the disease, regardless of the parasite presence or intensity, with macrophage hyperplasia, hemosiderosis, and emperipolesis being the main medullary changes in these animals. Also, the bone marrow of the femur and sternal manubrium are useful anatomical sites for the diagnosis of CVL by direct methods. Keywords: amastigotes; canine visceral leishmaniasis; histopathology; immunostaining; medullary change

Metabolic Peculiarities of Paracoccidioides brasiliensis Dimorphism as Demonstrated by iTRAQ Labeling Proteomics

Paracoccidioidomycosis (PCM), a systemic mycosis with a high incidence in Latin America, is caused by thermodimorphic fungi of the Paracoccidioides genus. The contact with host occurs by the inhalation of conidia or mycelial propagules which once reaching the pulmonary alveoli differentiate into yeast cells. This transition process is vital in the pathogenesis of PCM allowing the fungus survival in the host. Thus, the present work performed a comparative proteome analysis of mycelia, mycelia-to-yeast transition, and yeast cells of Paracoccidioides brasiliensis. For that, tryptic peptides were labeled with iTRAQ and identified by LC–MS/MS and computational data analysis, which allowed the identification of 312 proteins differentially expressed in different morphological stages. Data showed that P. brasiliensis yeast cells preferentially employ aerobic beta-oxidation and the tricarboxylic acid cycle accompanied by oxidative phosphorylation for ATP production, in comparison to mycelia and the transition from mycelia-to-yeast cells. Furthermore, yeast cells show a metabolic reprogramming in amino acid metabolism and in the induction of virulence determinants and heat shock proteins allowing adaptation to environmental conditions during the increase of the temperature. In opposite of that, the alcoholic fermentation found to P. lutzii, at least under laboratory conditions, is strongly favored in mycelium compared to yeast cells. Thereby, the data strongly support substantial metabolic differences among members of the Paracoccidioides complex, when comparing the saprobiotic mycelia and the yeast parasitic phases

New Methylcitrate Synthase Inhibitor Induces Proteolysis, Lipid Degradation and Pyruvate Excretion in <i>Paracoccidioides brasiliensis</i>

Background: Paracoccidioidomycosis is a systemic mycosis caused by the inhalation of conidia of the genus Paracoccidioides. During the infectious process, fungal cells use several carbon sources, leading to the production of propionyl-CoA. The latter is metabolized by the methylcitrate synthase, a key enzyme of the methylcitrate cycle. We identified an inhibitor compound (ZINC08964784) that showed antifungal activity against P. brasiliensis. Methods: This work aimed to understand the fungal metabolic response of P. brasiliensis cells exposed to ZINC08964784 through a proteomics approach. We used a glucose-free medium supplemented with propionate in order to simulate the environment found by the pathogen during the infection. We performed pyruvate dosage, proteolytic assay, dosage of intracellular lipids and quantification of reactive oxygen species in order to validate the proteomic results. Results: The proteomic analysis indicated that the fungal cells undergo a metabolic shift due to the inhibition of the methylcitrate cycle and the generation of reactive species. Proteolytic enzymes were induced, driving amino acids into degradation for energy production. In addition, glycolysis and the citric acid cycle were down-regulated while ß-oxidation was up-regulated. The accumulation of pyruvate and propionyl-CoA led the cells to a state of oxidative stress in the presence of ZINC08964784. Conclusions: The inhibition of methylcitrate synthase caused by the compound promoted a metabolic shift in P. brasiliensis damaging energy production and generating oxidative stress. Hence, the compound is a promising alternative for developing new strategies of therapies against paracoccidioidomycosis

Predicting copper-, iron-, and zinc-binding proteins in pathogenic species of the Paracoccidioides genus

Submitted by Jaqueline Silva ([email protected]) on 2018-05-08T12:14:22Z No. of bitstreams: 2 Artigo - Gabriel Brum Tristão - 2015.pdf: 2472227 bytes, checksum: 5bdf07b703f03b64021e19f11b883f4f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira ([email protected]) on 2018-05-09T11:51:54Z (GMT) No. of bitstreams: 2 Artigo - Gabriel Brum Tristão - 2015.pdf: 2472227 bytes, checksum: 5bdf07b703f03b64021e19f11b883f4f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Rejected by Luciana Ferreira ([email protected]), reason: Checar os nomes dos autores no lattes , alterar o país e rever os espaços na citação. on 2018-05-09T12:11:26Z (GMT)Submitted by Jaqueline Silva ([email protected]) on 2018-05-14T18:37:34Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Artigo - Gabriel Brum Tristão - 2015.pdf: 2472227 bytes, checksum: 5bdf07b703f03b64021e19f11b883f4f (MD5)Rejected by Luciana Ferreira ([email protected]), reason: O país não é Brasil. A citação apresenta problemas de espaço: TRISTÂO, Gabriel B.et al. Predicting copper-, iron-, and zinc-binding proteins in pathogenic species of the Paracoccidioides genus. Frontiers in Microbiology, Lausanne, v.(ESPAÇO)5, n.(ESPAÇO)761, p.(ESPAÇO)1-11, jan. 2015. on 2018-05-15T13:11:50Z (GMT)Submitted by Jaqueline Silva ([email protected]) on 2018-05-15T19:30:42Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Artigo - Gabriel Brum Tristão - 2015.pdf: 2472227 bytes, checksum: 5bdf07b703f03b64021e19f11b883f4f (MD5)Rejected by Luciana Ferreira ([email protected]), reason: Olhe a NBR 6023 como se abrevia os meses em outros idiomas e a questão de espaço depois de v. e n. (semre deice um espaço para depois digitar o número: ERRADO Tv.5, n.761, p.1-11, jan. 2015. CERTO v. 5, n. 761, p.1-11, Jan. 2015. on 2018-05-16T11:42:00Z (GMT)Submitted by Jaqueline Silva ([email protected]) on 2018-05-23T19:24:50Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Artigo - Gabriel Brum Tristão - 2015.pdf: 2472227 bytes, checksum: 5bdf07b703f03b64021e19f11b883f4f (MD5)Approved for entry into archive by Luciana Ferreira ([email protected]) on 2018-05-28T12:07:21Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Artigo - Gabriel Brum Tristão - 2015.pdf: 2472227 bytes, checksum: 5bdf07b703f03b64021e19f11b883f4f (MD5)Made available in DSpace on 2018-05-28T12:07:21Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Artigo - Gabriel Brum Tristão - 2015.pdf: 2472227 bytes, checksum: 5bdf07b703f03b64021e19f11b883f4f (MD5) Previous issue date: 2015-01Approximately one-third of all proteins have been estimated to contain at least one metal cofactor, and these proteins are referred to as metalloproteins. These represent one of the most diverse classes of proteins, containing metal ions that bind to specific sites to perform catalytic, regulatory and structural functions. Bioinformatic tools have been developed to predict metalloproteins encoded by an organism based only on its genome sequence. Its function and the type of metal binder can also be predicted via a bioinformatics approach. Paracoccidioides complex includes termodimorphic pathogenic fungi that are found as saprobic mycelia in the environment and as yeast, the parasitic form, in host tissues. They are the etiologic agents of Paracoccidioidomycosis, a prevalent systemic mycosis in Latin America. Many metalloproteins are important for the virulence of several pathogenic microorganisms. Accordingly, the present work aimed to predict the copper, iron and zinc proteins encoded by the genomes of three phylogenetic species of Paracoccidioides (Pb01, Pb03, and Pb18). The metalloproteins were identified using bioinformatics approaches based on structure, annotation and domains. Cu-, Fe-, and Zn-binding proteins represent 7% of the total proteins encoded by Paracoccidioides spp. genomes. Zinc proteins were the most abundant metalloproteins, representing 5.7% of the fungus proteome, whereas copper and iron proteins represent 0.3 and 1.2%, respectively. Functional classification revealed that metalloproteins are related to many cellular processes. Furthermore, it was observed that many of these metalloproteins serve as virulence factors in the biology of the fungus. Thus, it is concluded that the Cu, Fe, and Zn metalloproteomes of the Paracoccidioides spp. are of the utmost importance for the biology and virulence of these particular human pathogens

Popularization of Science: Demystifying the Central Dogma of Molecular Biology

This extension project was presented during the XII edition of the Cultural and Scientific Movement of Aparecida de Goiânia, Goiás, Brazil (MOCCA). The objective was to promote the basic scientific knowledge on the flow of genetic information from experimentation by illustrative methods. Each participant answered a questionnaire to assess their prior knowledge of the topic. He was then directed to a monitor that accompanied him, through the interactive environment, presenting illustrations with models, panels, three-dimensional models, as well as the accomplishment of the simple extraction of DNA from a fruit. At the end, the same questionnaire was reapplied. The comparative analysis of the participant’s performance in the questionnaire before and after the activities showed that interactive and illustrative practical approaches contributed positively to the better understanding of the content suggesting that this methodology can be incorporated into the teaching-learning process