Bovine papillomavirus isolation by ultracentrifugation.

The bovine papillomavirus (BPV) is the etiological agent of bovine papillomatosis, which causes significant economic losses to livestock, characterized by the presence of papillomas that regress spontaneously or persist and progress to malignancy. Currently, there are 13 types of BPVs described in the literature as well as 32 putative new types. This study aimed to isolate viral particles of BPV from skin papillomas, using a novel viral isolation method. The virus types were previously identified with new primers designed. 77 cutaneous papilloma samples of 27 animals, Simmental breed, were surgically removed. The DNA was extracted and subjected to PCR using Delta-Epsilon and Xi primers. The bands were purified and sequenced. The sequences were analyzed using software and compared to the GenBank database, by BLAST tool. The viral typing showed a prevalence of BPV-2 in 81.81% of samples. It was also detected the presence of the putative new virus type BR/UEL2 in one sample. Virus isolation was performed by ultracentrifugation in a single density of cesium chloride. The method of virus isolation is less laborious than those previously described, allowing the isolation of complete virus particles of BPV-2.

Bovine papillomavirus in beef cattle: first description of BPV-12 and putative type BAPV8 in Brazil.

Bovine papillomavirus (BPV) is an oncogenic virus associated with benign and malignant lesions, which result in notable economic losses. Peripheral blood samples and cutaneous papillomas were obtained from four adult beef cattle. Viral molecular identification was performed using specific primers for BPV-1, -2 and -4 in blood diagnosis and FAP59/FAP64 for skin papillomas. Histopathologic examination was done as a complementary and differential diagnosis. The fragments were purified, sequenced, and compared using BLASTn. The blood diagnosis showed the presence of BPV-2 and the analysis of cutaneous papillomas showed the presence of BPV-4, a new putative virus type BAPV8, and BPV-12, revealing for the first time the presence of BPV-12 and the putative type BAPV8 in beef cattle in Brazil. The sequences were deposited in the GenBank. Histopathology revealed acanthosis, hyperkeratosis, and koilocytosis in all samples analyzed. The presence of BAPV8 and BPV-12 in Brazil emphasizes the ubiquitous dissemination of BPVs in the herds of Brazil.

Phylogenetic classification and clinical aspects of a new putative Deltapapillomavirus associated with skin lesions in cattle.

Bovine papillomaviruses (BPVs) are recognized as causal agents of benign and malignant tumors in cattle. Thirteen types of BPVs have already been described and classified into 3 distinct genera. Divergences in the nucleotide sequence of the L1 gene are used to identify new viral types through the employment of PCR assays with degenerated primers. In the present study, a method for identifying BPVs based on PCR-RFLP and DNA sequencing allowed the identification of a new putative Deltapapillomavirus, designated JN/3SP (JQ280500.1). The analysis of the L1 gene showed that this strain was most closely related to the BPVs -1, -2, -13 , and OaPV1 (71-73% genetic similarity). In this study, we describe the detection of this new putative Deltapapillomavirus type and verify its phylogenetic position within the genus.

Bovine papillomavirus clastogenic effect analyzed in comet assay.

Bovine papillomavirus (BPV) is an oncogenic virus related to serious livestock diseases. Oncoproteins encoded by BPV are involved in several steps of cellular transformation and have been reported as presenting clastogenic effects in peripheral lymphocytes and primary culture cells. The aim of this study was to evaluate the clastogenic potential of BPV types 1, 2, and 4 by comet assay. Peripheral blood was collected from 37 bovines, 32 infected with different levels of papillomatosis (12 animals have no affection) and five calves, virus free (negative control). The viral identification showed presence of more than one virus type in 59.375% of the infected animals. Comet assay was performed according to alkaline technique. The Kruskal-Wallis test showed statistical difference between the negative control group and infected animals (P = 0.0015). The Dunn post hoc test showed difference comparing the infected animals with calves. Mann-Whitney U test verified no difference between animals infected with only one viral type and animals presenting more than one viral type. The comet assay is considered an efficient tool for assessment of damage in the host chromatin due to viral action, specifically highlighting viral activity in blood cells.

Cytogenetic studies in peripheral blood of bovines afflicted by papillomatosis.

Ten types of bovine papillomavirus (BPV) have been described and there are reports of viral transmission via blood. The presence of viral DNA in lymphocytes was described to be associated with chromosome instability in these cells. This study presents an evaluation of chromosome instability in short-term peripheral lymphocyte cultures from cows presenting skin papillomatosis, compared with asymptomatic infected animals and non-infected healthy bovines. In a total of 2203 cells, 918 (42%) showed at least one chromosome aberration: 42.7 (± 7.8) in animals with papillomatosis (BPV + W), 40.2 (± 11) in asymptomatic animals (BPV-W) and 4 (± 2) in control animals. Significant differences were found between the infected group (with or without symptoms) and the control group (P < 0.0001). The increased frequencies of chromosome aberrations suggest an interaction between the virus and host cell chromatin.

Simultaneous presence of bovine papillomavirus in blood and in short-term lymphocyte cultures from dairy cattle in Pernambuco, Brazil.

Bovine papillomaviruses (BPV) are the causal agents of benign and malignant lesions; they can cause dramatic economic losses in cattle. Although 10 virus types have been described, three types are most common in tumors, namely BPV-1, -2 and -4. Previous studies have reported BPV in blood cells and the possibility of blood acting as a latent virus site and/or transmission agent of virus dissemination. We studied a Holstein dairy herd in Pernambuco, Brazil, in which several animals showed severe cutaneous papillomatosis, without previous determination of BPV types. Blood samples and short-term lymphocyte cultures were collected from 54 cows. We compared the BPV types detected in peripheral blood to those identified in the respective lymphocyte cultures: BPV-1 was detected in 74% and BPV-2 in 87% of the whole blood samples. Simultaneous virus presence (BPV-1 and BPV-2) was found in 65% of the blood samples. BPV-1 or BPV-2 were detected in the lymphocyte cultures in 93% of the samples, and both in 89%. The detection of viral DNA in whole blood and in lymphocyte cultures is evidence that this virus is carried by lymphocytes.

[Care of women victims of spouse violence from the Heidegger's perspective of the technology world]. / Reflexões sobre o cuidar de mulheres que sofrem violência conjugal em uma perspectiva Heideggeriana do mundo da técnica.

This is concerned with a reflexive text that approaches the subject of the care to women that suffer conjugal violence in an analytic perspective of the world of the technique, developed from conceptions and concepts of Martin Heidegger, contemporary philosopher of phenomenology. The care of women that suffer conjugal violence appears as one that considers values, decision, trust to care, knowledge and care actions and its consequences.

[Psychodrama as a pedagogical strategy: experiences in undergraduate teaching of women's health services]. / Psicodrama como estratégia pedagógica: vivências no ensino de graduação na área de saúde da mulher.

As the result from a discussion by the Grupo de Estudos sobre Saúde da Mulher--GEM (Group of Studies on Women's Health), psychodrama has been used as a pedagogical strategy to develop the topic Female's Conditions and Women's Health. Written and oral individual accounts as well as stories constructed in "group meetings" were treated by the gender perspective. The dynamics has facilitated relationships and self-knowledge, making it possible for day-by-day situations to emerge. The reflection between what is lived and what is theoretical has allowed the visibility of women's insertion in the public and private sectors as well as created situations for the construction of concepts.

[Development of a milk beverage based on pumpkin flakes]. / Desenvolvimento de bebida láctea a base de flocos de abóbora.

Vitamin A deficiency is one of the majors public health problems in Brazilian Northeast, and among other causes may be concerned to precocious weaning. Aiming at reducing this problem, a dehydrated product based on powdered milk and pumpkin flakes was developed to act as a carotene source at weaning period. Pumpkin flakes were obtained by drum drying at a 6 atm steam pressure, 0.75 m2 contact surface and 1 rpm, and had their content of carotenoids, beta carotene, centesimal composition molds and yeast and faecal coliforms evaluated. The flakes were added to sugar and whole powdered milk according to FAO/WHO nutritional recommendation to 6-12 months old children and submitted to acceptability test by a 6 judges' panel. The results showed that the drying process had a 7% efficiency score and the flakes composition presented 4.84% moisture; 4.0% protein; 5.5% ash; 1.30% fat; 6.22% fiber; 78.14% carbohydrates; 115.08 micrograms/g total carotenois and 80.64 micrograms/g beta-carotene content. No coliforms were detected and molds and yeast content was 4.0 x 10(2) CFU/g. The powdered formulation properly diluted in water supports 45% of the RDA for children (calories, protein, carbohydrates and lipids) and 100% vitamin A and protein considering a 400 ml/day ingestion.