Identificação e validação de genes diferencialmente expressos em carcinoma de pênis

O carcinoma de pênis (CEP) é um tumor epitelial invasivo raro com alta morbidade decorrente da própria doença e/ou de seu tratamento. O perfil socioeconômico e cultural dos pacientes dificulta o diagnóstico precoce, tratament o e seguimento dos enfermos. Pacientes sem tratamento geralmente morrem dentro de dois anos após diagnóstico devido à proliferação celular regional ou a metástases distantes. Não há padrão nos sistemas de estadiamento e nas condutas clínicas, resultando em dificuldades na abordagem terapêutica. Comparado ao câncer cervical, poucos estudos moleculares em CEP foram realizados. O presente projeto teve como objetivo geral identificar genes diferencialmente expressos em tecidos penianos tumorais e normais e o possível papel do vírus do papiloma humano (HPV) no desenvolvimento de CEP. Diferenças na expressão gênica entre tecidos tumorais e normais foram verificadas pela metodologia de RaSH (rapid subctration hybridization), que selecionou 5 genes possivelmente rel acionados à carcinogênese ( KIAA1033, NAMPT, RPL6, CDKN2A e ANXA1) com expressão alterada em tecidos tumorais. A validação das alterações de expressão destes genes foi realizada por polymerase chain reaction (reação em cadeia da polimerase, PCR) em tempo real. Tanto os genes como as proteínas de ANXA1 como CDKN2A apresentaram aumento de expressão em pacientes com presença de HPV de alto risco em comparação com tecidos negativos para HPV. Por imunohistoquímica foi possível estabelecer correlação entre alterações na expressão da proteína anexina A1 e a presença de HPV de alto risco. Pacientes portadores de CEP provenientes de duas regiões (São Paulo e Pará) foram avaliados para a presença de HPV por PCR convencional. Amostras positivas foram genotipadas por hib ridização...Penile carcinoma is a rare epithelial tumor with high morbidity due to own disease and/or treatment. The socio -economic and cultural profile of the patients hampers early diagnosis, treatment and follow-up. Without treat ment patients usually dies within two years after first diagnosis due to regional cellular proliferation or distant metastasis. There is no standard in staging systems or in clinical procedures, resulting in difficulties in the therapeutic approach. Compar ed to cervical cancer, there are few molecular studies about this tumor. The present work aimed to identify differentially expressed genes in tumor and normal penile tissues and the role of human papillomavirus (HPV) in the development of this tumor. Differences in gene expression between tumoral and normal tissues were accessed by RaSH (rapid subctration hybridization) methodology, which selected five possibly carcinogenesis related genes (KIAA1033, NAMPT, RPL6, CDKN2A and ANXA1) with altered expression in tumor tissues. The validation of gene expression changes was performed by real time polymerase chain reaction (PCR). The ANXA1 and CDKN2A, both genes and proteins, showed superexpression in patients with high-risk HPV compared to HPV negative tissues. Immunohistochemical assays established a correlation between alterations in the superexpression of annexin A1 and the presence of high risk HPV. Penile cancer harboring patients from two regions (São Paulo and Pará) was assessed for HPV presence by conventional PCR. Positive samples were genotyped by reverse hybridization-based line probe assay (INNO -LiPA). The overall HPV prevalence in single or multiple infections was significantly greater in Pará (81.67%) than in São Paulo (64.10%). A wilder range of genotypes were found in Pará, suggesting that the genotypes circulating in the population of Pará... (Complete abstract click electronic access below)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Mosquito-transmitted viruses - the great Brazilian challenge

ABSTRACT Arboviruses pose a serious threat to public health worldwide, overloading the healthcare system and causing economic losses. These viruses form a very diverse group, and in Brazil, arboviruses belonging to the families Flaviviridae and Togaviridae are predominant. Unfortunately, the number of arboviruses increases in proportion with factors such as deforestation, poor sanitation, climate changes, and introduction of new viruses like Chikungunya virus and Zika virus. In Brazil, dengue is endemic, along with the presence of other arboviruses. The situation is complicated by the scarcity of diagnostic infrastructure and the absence of approved vaccines for these diseases. Disease control, thus, relies solely on vector control. Therefore, enhanced clinical knowledge and improved general awareness about these arboviruses are indispensable to tackle diagnostic inadequacies

Mayaro fever in an HIV-infected patient suspected of having Chikungunya fever

Abstract Arboviruses impose a serious threat to public health services. We report a case of a patient returning from a work trip to the Amazon basin with myalgia, arthralgia, fever, and headache. During this travel, the patient visited riverside communities. Both dengue and Chikungunya fevers were first suspected, tested for, and excluded. Mayaro fever was then confirmed by reverse transcription polymerase chain reaction followed by next-generation sequencing and phylogenetic reconstruction. The increased awareness of physicians and consequent detection of Mayaro virus in this case was only possible due a previous surveillance program with specific health personnel training about these neglected arboviruses

Mayaro fever in an HIV-infected patient suspected of having Chikungunya fever

Abstract Arboviruses impose a serious threat to public health services. We report a case of a patient returning from a work trip to the Amazon basin with myalgia, arthralgia, fever, and headache. During this travel, the patient visited riverside communities. Both dengue and Chikungunya fevers were first suspected, tested for, and excluded. Mayaro fever was then confirmed by reverse transcription polymerase chain reaction followed by next-generation sequencing and phylogenetic reconstruction. The increased awareness of physicians and consequent detection of Mayaro virus in this case was only possible due a previous surveillance program with specific health personnel training about these neglected arboviruses

Lipid Droplets as Potential Antiviral Targets for Flaviviruses

Lipid droplets (LDs) are intracellular sites for neutral lipid storagecritical for lipid metabolism and energy homeostasis. Structurally, LDshave a rich lipid content but they also contain several functionally diversetypes of proteins, including already described pro and antiviral relevantfactors. New roles for LDs in immunity have been uncovered, withevidence that they act as assembly and replication platforms for specificviruses and as reservoirs for proteins that counteract intracellularpathogens. LDs might also have potential implications for mechanisms ofcell proliferation and differentiation, affecting viral replication. Cellpathological conditions have been noticed to share an altered lipogenesisphenotype. However, the mechanisms that regulate LDs formation orconsumption and how their function impacts the cellular biology remainunknown. These evidences led us to investigate the effect of the alterationof LDs abundance during the replication of mosquito-transmittedflaviviruses. Our in vitro studies show that there is a decreased LDscontent in dengue, Zika and yellow fever infected cells, demonstratingthat these structures might be essential for diverse RNA virus life cycleand could be considered a novel antiviral target.Fil: Vázquez, Cecilia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Giovannoni, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Irene Bosch. Massachusetts Institute of Technology; Estados UnidosFil: de Oliveira Mota, Mânlio Tasso. Faculty of Medicine of São José do Rio Preto; BrasilFil: Lacerda Nogueira, Mauricio. Faculty of Medicine of São José do Rio Preto; BrasilFil: Cordo, Sandra Myriam. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Garcia, Cybele. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentin

Incidence of HPV infection in the uterine cervix in HIV positive pregnant women seen at the Hospital de Base in São José do Rio Preto, Brazil

Objective: To evaluate the incidence of HPV-HIV co-infection among pregnant women and to relate it to CD4+ cell count in peripheral blood, to the  expression of protein p16 in the epithelium of the uterine cervix and  to the presence of HPV DNA in cervical biopsies. Methods: Pregnant women at the same age were selected and distributed into two groups with 70 patients  each: HIV positive (Study Group) and HIV negative (Control I). Another group (Control II) comprised 36 HIV negative patients at different ages and with the diagnosis of cervical intraepithelial neoplasia, in the period from 2000 to 2007. Colpocytology and/or cervical biopsy of these patients were analyzed for the occurrence of morphological characteristics, suggesting HPV infection; for the CD4 count in peripheral blood; and expression of protein p16 in the cervix epithelium and compared the findings with p16 expression in a group of 36 women with cervical epithelial neoplasm. PCR was performed in the samples of the Study Group to confirm the presence of HPV DNA. Rresults: The 70 HIV positive pregnant patients were aged from 15 to 45 years (mean of  28 years, median of 28.2 years). Twenty-two (31.4%) of them presented morphological changes consistent with cervical HPV infection; of these, 16 presented CD4 count lower than 500 cells (p=0.03). In the  HIV negative  Group, one  patient (1.4%) had cervical intraepithelial lesion. Ten cervical biopsies  in the Study  Group presented HPV DNA in the PCR. The Control Group II showed similar results to the Study Group. Cconclusions: The incidence of HPV-HIV co-infection in pregnant women was relevant compared to  the HIV negative Group. In the HIV-positive Group, the incidence of HPV was directly related to low immunity. The p16 expression was stronger among the cases diagnosed as having higher grade intraepithelial cervical lesions (CIN 2 and 3). The presence of HPV DNA was confirmed through PCR in cervical lesions of HIV positive pregnant women. This study points to the importance of having periodic gynecological exams and routine cytology screening tests for the early detection of HPV infection during antenatal care of HIV-positive pregnant women