Phylogenetic and structural studies of a novel equine papillomavirus identified from aural plaques

Papillomaviruses (PVs) infect a wide range of animal species and show great genetic diversity. To date, excluding equine sarcoids, only three species of PVs were identified associated with lesions in horses: Equus caballus papillomavirus 1 (EcPV1-cutaneous), EcPV2 (genital) and EcPV3 (aural plaques). In this study, we identified a novel equine PV from aural plaques, which we designated EcPV4. Cutaneous samples from horses with lesions that were microscopically diagnosed as aural plaques were subjected to DNA extraction, amplification and sequencing. Rolling circle amplification and inverse PCR with specific primers confirmed the presence of an approximately 8. kb circular genome. The full-length EcPV4 L1 major capsid protein sequence has 1488 nucleotides (495 amino acids). EcPV4 had a sequence identity of only 53.3%, 60.2% and 51.7% when compared with the published sequences for EcPV1, EcPV2 and EcPV3, respectively. A Bayesian phylogenetic analysis indicated that EcPV4 clusters with EcPV2, but not with EcPV1 and EcPV3. Using the current PV classification system that is based on the nucleotide sequence of L1, we could not define the genus of the newly identified virus. Therefore, a structural analysis of the L1 protein was carried out to aid in this classification because EcPV4 cause lesion similar to the lesion caused by EcPV3. A comparison of the superficial loops demonstrated a distinct amino acid conservation pattern between EcPV4/EcPV2 and EcPV4/EcPV3. These results demonstrate the presence of a new equine PV species and that structural studies could be useful in the classification of PVs. © 2012 Elsevier B.V.

CDKN2A promoter hypermethylation in astrocytomas is associated with age and sex

CDKN2A promoter hypermethylation has been widely related to many cancers. In astrocytomas, although CDKN2A (p16INK4A protein) is often inactivated, there are still some controversial issues regarding the mechanism by which this alteration occurs. Thus, we analyzed a series of astrocytomas to assess the association between CDKN2A expression and methylation of grade I-IV tumors (WHO) and clinicopathological parameters. DNA extracted from formalin-fixed paraffin-embedded material of 93 astrocytic tumors was available for CDKN2A promoter methylation analysis and p16INK4A expression by methylation-specific PCR and immunohistochemistry, respectively. A strong negative correlation between nuclear and cytoplasmic immunostaining and CDKN2A promoter methylation was found. Additionally, a significant negative correlation between CDKN2A promoter methylation and age was observed; also, female patients had statistically more CDKN2A methylated promoters (p=0.036) than men. In conclusion, CDKN2A inactivation by promoter methylation is a frequent event in astrocytomas and it is related to the age and sex of patients. © 2013 Surgical Associates Ltd.

Description of three new species of Hepatozoon (Apicomplexa, Hepatozoidae) from Rattlesnakes (Crotalus durissus terrificus) based on molecular, morphometric and morphologic characters

Hepatozoon spp. are commonly found infecting snakes. Since the latter are parasitized by diverse forms and data in the literature show divergence, we studied Hepatozoon spp. diversity on Crotalus durissus terrificus snakes using both molecular and morphological approaches. Naturally infected animals were employed. Blood was collected, blood smears were prepared and an aliquot was stored at -20. °C for DNA extraction. Five specimens of C. durissus terrificus were selected, each of them infected with one gamont type. Morphological and morphometric analyses of the found gamonts led to their grouping into three populations. For molecular characterization, seven oligonucleotide pairs that amplify distinct regions of rDNA gene were tested by adopting the PCR technique. Only the oligonucleotide pairs HepF300/Hep900 and HEMO1/HEMO2 were efficient in amplifying and distinguishing different isolates of Hepatozoon spp. from snakes. The better results were obtained when both oligonucleotide pairs were used in association. Based on the molecular and morphologic differences, three new species were proposed: Hepatozoon cuestensis sp. nov.; Hepatozoon cevapii sp. nov. and Hepatozoon massardii sp. nov. This is the first description of new Hepatozoon species from snakes, based on molecular characterization and morphological data, in South America. © 2013 Elsevier Inc.

An HPLC-UV method for the quantification of zidovudine in rat plasma

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Método simplificado de extração e quantificação do herbicida tebuthiuron em solo sob diferentes sistemas de cultivo de cana-de-açúcar

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Detecção do gene de peroxidase em sementes de soja pela reação da polimerase em cadeia (PCR)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da disponibilidade de boro em um latossolo de Mato Grosso/MT

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Metodologia para avaliação de rodovias extraídas computacionalmente em imagens digitais

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ocorrência de infecções por Babesia spp. e Hepatozoon spp. em gatos domésticos (Felis domesticus) do Estado de São Paulo e do Distrito Federal

Pós-graduação em Medicina Veterinária - FMVZ

Estudo quantitativo da infecção por Babesia bovis e Babesia bigemina em diferentes grupos genéticos de bovinos de corte

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Polimorfismo nos genes da leptina e do receptor de melatonina em búfalas (Bubalus bubalis)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Marcadores moleculares associados à qualidade de carne em bovinos Nelore

Pós-graduação em Genética e Melhoramento Animal - FCAV

Padronização de um protocolo para detecção molecular de Leptospira spp. e Brucella spp. em sêmen bovino comercial

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ocorrência de crryptosporidium spp, em animais exóticos de companhia no Brasil: Milena Sato de Souza. -

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação física, epidemiológica e molecular da infecção por Cryptosporidium spp. em passeriformes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)