Abstract Background Merkel cell polyomavirus (MCPyV), a human polyomavirus that is unequivocally linked to merkel cell carcinoma (MCC), has been found in association with keratinocytes carcinomas (KC), especially basal cell carcinoma (BCC) and cutaneous squamous cell carcinoma (cSCC). Nevertheless, there is scarce information about the possible involvement of MCPyV in the development of KC. Objectives To assess the presence of MCPyV DNA and Large-T Antigen (LT-Ag) via Polymerase Chain Reaction (PCR) and Immunohistochemistry (IHC) in cases of KC, and to correlate its presence with immunohistochemical markers p16, p53, and ki67, tumor type and subtype, sun-exposed location, and epidemiological data. Methods The prevalence of MCPyV DNA, LT-Ag, and immunohistochemical markers p16, p53, and ki67 was assessed by PCR and Immunohistochemistry (IHC) in 127 cases of KC, these results were correlated with tumor type and subtype, sun-exposed location, and epidemiological data. Results The MCPyV DNA was detected in 42.57% (43 of 101) cases by PCR, the LT-Ag was detected in 16.4% (20 of 122) of cases, p16 in 81.5% (97 of 119), p53 in 66.4% (83 of 125), ki67 in 89% (73 of 82). No correlation between MCPyV LT-Ag and DNA confronted with tumor type, subtype, location site, and immunohistochemical markers was found. A single correlation between the MCPyV LT-Ag and cSCC tumors and peri-tumoral lymphocyte cells was noted. Study limitations Further steps need to be taken to better evaluate the MCPyV influence and its possible role in KC carcinogenesis, as the evaluation of the virus genome state, the gene sequence that encodes LT-Ag in the KC tumor cells, and in situ hybridization for viral DNA or RNA in these cells. Conclusions Despite the frequent detection of MCPyV in KC, the data available so far does not support the hypothesis of a causal relationship between them. MCPyV, , (MCPyV) MCC, MCC (MCC) (KC) BCC (BCC cSCC. . (cSCC) Nevertheless LargeT Large T LTAg LT Ag (LT-Ag (PCR IHC (IHC p ki subtype sunexposed sun exposed LTAg, Ag, ki6 12 4257 42 57 42.57 43 (4 101 164 16 4 16.4 20 (2 122 p1 815 81 5 81.5 97 (9 119, 119 119) p5 664 66 66.4 83 (8 125, 125 125) 89 73 (7 82. 82 82) site peritumoral peri tumoral noted carcinogenesis state them (MCPyV (MCC (KC (cSCC 1 425 42.5 ( 10 16. 2 8 81. 9 11 6 66. 7 42.

Abstract Viruses have been frequently identified in several human neoplasms, but the etiological role of these viruses in some tumors is still a matter of controversy. Polyomaviruses stand out among the main viruses with oncogenic capacity, specifically the Merkel cell polyomavirus (MCPyV). Recent revisions in the taxonomy of polyomaviruses have divided the Polyomaviridae family into six genera, including 117 species, with a total of 14 currently known human-infecting species. Although the oncogenicity of polyomaviruses has been widely reported in the literature since 1950, the first description of a polyomavirus as an etiological agent of a neoplasm in humans was made only in 2008 with the description of MCPyV, present in approximately 80% of cases of Merkel cell carcinoma (MCC), with the integration of its genome to that of the tumor cells and tumor-specific mutations, and it is considered the etiological agent of this neoplasm since then. MCPyV has also been detected in keratinocyte carcinomas, such as basal cell carcinoma and squamous cell carcinoma of the skin in individuals with and without immunosuppression. Data on the occurrence of oncogenic viruses potentially involved in oncogenesis, which cause persistence and tissue injury, related to the Merkel cell polyomavirus are still scarce, and the hypothesis that the Merkel cell polyomavirus may play a relevant role in the genesis of other cutaneous carcinomas in addition to MCC remains debatable. Therefore, the present study proposes to explore the current knowledge about the presence of MCPyV in keratinocyte carcinomas. neoplasms controversy capacity MCPyV. . (MCPyV) genera 11 species 1 humaninfecting infecting 1950 200 80 MCC, , (MCC) tumorspecific specific mutations then immunosuppression oncogenesis injury scarce debatable Therefore (MCPyV 195 20 8 (MCC 19 2

ABSTRACT BKV and JCV belong to the Polyomaviridae family and are opportunistic agents associated with complications in immunocompromised individuals. Although a single screening assay for both viruses would be convenient, the diversity of BKV and JCV serotypes and genotypes is a methodological challenge. In this paper, we developed a PCR method able to detect and segregate BKV and JCV, despite these genetic discrepancies. A duplex semi-nested PCR (duplex snPCR) was designed to target a conserved region (639nt-1516nt) within the VP2 gene. In the first PCR, a primer set common to all BKV and JCV serotypes/ genotypes was used, followed by a semi-nested PCR with internal primers for BKV and JCV segregation. The limit of detection of the duplex snPCR was as low as 10 copies of BKV or JCV plasmids/μL. Specific products were observed when JCV and BKV plasmids were mixed in the same reaction. In field sample testing, the duplex snPCR detected and distinguished both viruses in different biological samples. Results were confirmed by Sanger's sequencing. The geographical complexity of BKV and JCV serotypes and genotypes imposes limits to a simple and universal method that could detect each virus. However, we describe here a sensitive and reliable PCR technique for BKV and JCV diagnosis that overcomes these limitations and could be universally applied.

BK Poliomavírus (BKPyV) é um agente causal de nefropatia, estenose ureteral e cistite hemorrágica em receptores de transplante renal, sendo considerado uma importante doença emergente na transplantação. Rastreamento regular para reativação do BKPyV, principalmente nos dois primeiros anos pós-transplante, com subsequente redução preemptiva da imunossupressão é considerada a melhor conduta para evitar a progressão da doença, já que a eliminação ou redução da viremia é alcançada na grande maioria dos pacientes dentro de 6 meses. O uso de drogas com propriedades antivirais para os pacientes com viremia persistente tem sido tentado, embora sem benefícios claros. As manifestações clínicas da nefropatia por BKPyV, as estratégias para o diagnóstico e monitoramento da infecção por BKPyV, o manejo do regime de imunossupressão após a detecção da reativação do BKPyV e o uso de drogas antivirais são discutidas nesta revisão.

BK polyomavirus (BKPyV) is a causal agent of nephropathy, ureteral stenosis and hemorrhagic cystitis in kidney transplant recipients, and is considered an important emerging disease in transplantation. Regular screening for BKPyV reactivation mainly during the first 2 years posttransplant, with subsequent pre-emptive reduction of immunosuppression is considered the best option to avoid disease progression, since successful clearance or reduction of viremia is achieved in the vast majority of patients within 6 months. The use of drugs with antiviral properties for patients with persistent viremia has been attempted despite unclear benefits. Clinical manifestations of BKPyV nephropathy, current strategies for diagnosis and monitoring of BKPyV infection, management of immunosuppressive regimen after detection of BKPyV reactivation and the use of antiviral drugs are discussed in this review.

We investigated the occurrence of HIV-1 antiretroviral resistance in individuals failing to respond to highly active antiretroviral therapy (HAART) attended by RENAGENO from 2001-2004. One hundred and seventeen patients were selected for this study; their plasma viral RNA was extracted and the PR and RT genes sequenced to examine subtype, genetic polymorphisms and mutations associated with resistance to antiretroviral drugs. HIV-1 sequence analysis showed that 86/100 (86%) were infected with subtype B, 7/100 (7%) with subtype F and 7/100 (7%) with RT/PR hybrid forms (2 D/B, 2 F/B, 2 B/F and 1 D/F). In 14 (12%) of the samples, the subtype was not determined. The prevalence of resistance mutations was high (93.1%), mainly in the RT gene. The most prevalent resistance mutations were: M184V (60.7%), T215Y (49.6%) and M41L (46.7%) in the RT gene and L90M (19.6%), M46I (16.2%) and D30N (12.8%) in the PR gene. The frequency of resistance mutations tended to increase from the first to the second therapeutic scheme failure (p=0.079); but it stabilized after subsequent failures (p=0.875). Our finding of a high frequency of drug resistant HIV-1 samples supports the need for continuous genotypic monitoring of patients failing HAART.

Desde sua introdução na América do Norte em 1999, mais de 27.500 casos humanos da infecção por West Nile virus (WNV) foram reportados nos Estados Unidos da América (EUA), resultando em mais de 1000 casos fatais. Recentemente, a disseminação do vírus para o hemisfério sul foi confirmada com a detecção de animais infectados pelo WNV em território sul-americano. A soropositividade para WNV em eqüídeos na Colômbia e Venezuela e o isolamento do vírus nestes animais na Argentina, reiteram a necessidade da manutenção do sistema de vigilância enzoótica para WNV em território brasileiro. Aspectos pertinentes à infecção, patogenia e epidemiologia do WNV são discutidos neste artigo.

Since the West Nile virus (WNV) was introduced in North America in 1999, more than 27,500 cases were reported among humans in the US, resulting in more than 1,000 casualties. Recently, the dissemination of the WNV to the Southern Hemisphere was confirmed through the detection of seropositive animals. Positively-infected horses for WNV in Colombia, Venezuela and viral isolation in Argentina uphold the need to maintain the enzootic surveillance system in the Brazilian territory. Aspects related to infection, diagnosis and epidemiology of WNV are discussed in this article.

Desde sua introdução na América do Norte em 1999, mais de 27.500 casos humanos da infecção por West Nile virus (WNV) foram reportados nos Estados Unidos da América (EUA), resultando em mais de 1000 casos fatais. Recentemente, a disseminação do vírus para o hemisfério sul foi confirmada com a detecção de animais infectados pelo WNV em território sul-americano. A soropositividade para WNV em eqüídeos na Colômbia e Venezuela e o isolamento do vírus nestes animais na Argentina, reiteram a necessidade da manutenção do sistema de vigilância enzoótica para WNV em território brasileiro. Aspectos pertinentes à infecção, patogenia e epidemiologia do WNV são discutidos neste artigo.

Since the West Nile virus (WNV) was introduced in North America in 1999, more than 27,500 cases were reported among humans in the US, resulting in more than 1,000 casualties. Recently, the dissemination of the WNV to the Southern Hemisphere was confirmed through the detection of seropositive animals. Positively-infected horses for WNV in Colombia, Venezuela and viral isolation in Argentina uphold the need to maintain the enzootic surveillance system in the Brazilian territory. Aspects related to infection, diagnosis and epidemiology of WNV are discussed in this article.

Políticas de saúde adotadas pelo governo brasileiro têm resultado em uma estabilização e mesmo redução da Aids no país, muito embora esta tendência não venha acontecendo em municípios de menor porte. Neste contexto, analisamos o impacto da Aids no município de Teresópolis/RJ, com a finalidade de obter um perfil regional da epidemia como modelo para cidades com características demográficas similares. Para tal, comparamos diversos critérios epidemiológicos para a epidemia entre o município de Teresópolis, o Estado do Rio de Janeiro (RJ) e o Brasil, entre 2000-2004, bem como a incidência da doença em cidades do Estado do RJ entre 1996-2004, utilizando para isto informações provenientes dos bancos de dados do Sistema Nacional de Agravos Notificáveis (SINAN) e Instituto Brasileiro de Geografia e Estatística (IBGE). Os resultados indicam que a epidemia de Aids em Teresópolis apresenta um perfil semelhante ao estadual e nacional com pequenas variações, embora a incidência de Aids no município esteja elevada em relação aos demais municípios do Estado. Concluindo, apesar de o país tratar a questão da Aids de forma prioritária, é necessária uma investigação mais cuidadosa nos municípios de menor porte fora das regiões metropolitanas, de forma a controlar a tendência da epidemia de Aids nestas localidades.

The healthcare policies adopted by the Brazilian government resulted in the stabilization or even reduction of the Aids epidemic in the country, although the incidence of the disease seems to be increasing in smaller municipalities. In this context, the impact of Aids in Teresópolis/RJ was studied in order to obtain a regional profile of the epidemic as a model for demographically similar cities. To achieve this purpose, many epidemiological relevant aspects of Aids were compared between Teresópolis, the state of Rio de Janeiro (RJ) and Brazil from 2000-2004, as well as the incidences of the disease in many cities in RJ from 1996-2004 by using information from the Sistema Nacional de Agravos Notificáveis-SINAN (National Disease Reporting System) and Instituto Brasileiro de Geografia e Estatística-IBGE (Brazilian Institute of Geography and Statistics) databases. The results indicate that the Aids epidemic in Teresópolis is similar to that of the state of RJ and the country with small variations, although the incidence of Aids is still high in comparison to other municipalities of the state. In conclusion, although Aids is treated as a priority in the country, a careful investigation in small municipalities outside metropolitan areas is necessary in order to control Aids epidemic trends in such locations.

Políticas de saúde adotadas pelo governo brasileiro têm resultado em uma estabilização e mesmo redução da Aids no país, muito embora esta tendência não venha acontecendo em municípios de menor porte. Neste contexto, analisamos o impacto da Aids no município de Teresópolis/RJ, com a finalidade de obter um perfil regional da epidemia como modelo para cidades com características demográficas similares. Para tal, comparamos diversos critérios epidemiológicos para a epidemia entre o município de Teresópolis, o Estado do Rio de Janeiro (RJ) e o Brasil, entre 2000-2004, bem como a incidência da doença em cidades do Estado do RJ entre 1996-2004, utilizando para isto informações provenientes dos bancos de dados do Sistema Nacional de Agravos Notificáveis (SINAN) e Instituto Brasileiro de Geografia e Estatística (IBGE). Os resultados indicam que a epidemia de Aids em Teresópolis apresenta um perfil semelhante ao estadual e nacional com pequenas variações, embora a incidência de Aids no município esteja elevada em relação aos demais municípios do Estado. Concluindo, apesar de o país tratar a questão da Aids de forma prioritária, é necessária uma investigação mais cuidadosa nos municípios de menor porte fora das regiões metropolitanas, de forma a controlar a tendência da epidemia de Aids nestas localidades.

The healthcare policies adopted by the Brazilian government resulted in the stabilization or even reduction of the Aids epidemic in the country, although the incidence of the disease seems to be increasing in smaller municipalities. In this context, the impact of Aids in Teresópolis/RJ was studied in order to obtain a regional profile of the epidemic as a model for demographically similar cities. To achieve this purpose, many epidemiological relevant aspects of Aids were compared between Teresópolis, the state of Rio de Janeiro (RJ) and Brazil from 2000-2004, as well as the incidences of the disease in many cities in RJ from 1996-2004 by using information from the Sistema Nacional de Agravos Notificáveis-SINAN (National Disease Reporting System) and Instituto Brasileiro de Geografia e Estatística-IBGE (Brazilian Institute of Geography and Statistics) databases. The results indicate that the Aids epidemic in Teresópolis is similar to that of the state of RJ and the country with small variations, although the incidence of Aids is still high in comparison to other municipalities of the state. In conclusion, although Aids is treated as a priority in the country, a careful investigation in small municipalities outside metropolitan areas is necessary in order to control Aids epidemic trends in such locations.

Two different procedures for inoculation of HSV on corneas of BALB/c mice were evaluated. The first was by the use of HSV suspensions directly on the corneas and the other was after corneal scarification. Animals by this later method presented greater morbidity and mortality than those of first group, suggesting that inoculation of HSV without scarification of the cornea should be the method of choice for the study of HSV ophthalmic infection. This model showed also be an efficient experimental system to testing antiviral drugs.