Immunohistochemical evaluation of e-cadherin, Ki-67 and PCNA in canine mammary neoplasias: Correlation of prognostic factors and clinical outcome

E-cadherin is a cell-cell adhesion molecule and low e-cadherin expression is related to invasiveness and may indicate a bad prognosis in mammary neoplasms. The expression of cell proliferation markers PCNA and especially Ki-67, has also proved to have a strong prognostic value in this tumor class. The expression of these markers was related to the clinical-pathological characteristics of 73 surgically removed mammary tumors in female dogs by immunohistochemistry. There was no statistical correlation between these markers and death by neoplasm, survival time and disease-free interval. However, the loss of e-cadherin expression and marked Ki-67 expression (p=0.016) were considered statistically significant for the diagnosis (p=0.032). When evaluated as independent factors, there was evidence of the relationship between the loss of e-cadherin expression and high PCNA expression with changes in the body status (divided into obese, normal and cachectic) of female dogs (p=0.030); there was also evidence of the relationship between pseudopregnancy and e-cadherin alone (p=0.021) and for ulceration and PCNA alone (p=0.035). The significant correlation between the markers expression and these well known prognostic factors used individually or in combination suggests their prognostic value in canine mammary tumors.

Prevalência de papiloma vírus (HPV) perianal assintomático em pacientes portadores de HPV genital tratados no Hospital das Clínicas da Faculdade de Medicina de Botucatu

Introduction: Human Papillomavirus (HPV) infection is one of the Sexually Transmitted Diseases (STD) of greatest incidence and prevalence worldwide, and it is presently considered to be a pre-neoplastic lesion. Human Papillomavirus infection has totaled 23.4% of the sexually transmitted diseases reported to the Department of Health, and it is currently the most common in our country. Many patients are asymptomatic carriers. Methods: Twenty patients who had been previously treated for HPV due to genital lesions were referred for the conduction of anuscopy and colposcopy of the perianal region. Results: Males (80%) prevailed over females (20%). Of the total number of patients, only 2 showed lesions as examined by anuscopy (10%). However, 3 other patients showed lesions by means of colposcopy, thus increasing to 5 (25%) the total number of asymptomatic patients who presented perianal lesions. Of the total number of patients with lesions, 4 were males and 1 was a female. Conclusion: The presence of perianal lesions was observed in 10% of the patients with genital lesions by means of simple anal inspection. This figure increased to 25% when anal colposcopy was associated, thus showing the importance of conducting such examination on all patients with increased risk factors for HPV infection in the anal region.

Compensatory canine angulation in angle class II and III patients

Abstract: The aim of this study was to evaluate the occurence of compensation in mesiodistal axial inclinations of canines in skeletal malocclusions patients. The sample consisted of 25 Angle Class II, division 1 malocclusion (group 1) and 19 Angle Class III malocclusion patients (group 2). After measurement of dental angulations through a method that associates plaster model photography and AutoCad software, comparisons between the groups were performed by T-test for independent samples. Results showed that there was no statistically significant difference (p ≤ 0.05) between groups, when maxillary canine angulations were compared. Regarding the mandibular canines, there was a statistically significant difference in dental angulation, expressed by 3.2° for group 1 and 0.15° for group 2. An upright position tendency for mandibular canines was observed in the Angle Class III sample. This configures a pattern of compensatory coronary positioning, since the angulation of these teeth makes them occupy less space in the dental arch and consequently mandibular incisors can be in a more retracted position in the sagittal plane.

COX-2 and TGF-β expression in proliferative disorders of canine prostate

COX-2 and TGF-β expression was determined in order to correlate non-neoplastic lesions, preneoplastic lesions and carcinoma in the prostate of dogs. The results show that neoplastic and preneoplastic lesions express more COX-2 and TGF-β when compared to carcinomas, which suggests these proteins may cooperate in the process of prostate tumorigenesis.

Expression of pro-and-anti-apoptotic antigens in the cerebellum of dogs naturally infected with canine distemper virus

Canine distemper virus (CDV) may induce multifocal demyelination in the central nervous system of infected dogs. The present work investigated apoptosis in white and gray matter (granular layer) in the cerebellum of naturally infected dogs by the analysis of the expression of the pro-apoptotic antigens caspase - 2 and - 3, b(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL-staining) positivity, annexin-V immunodetection, and the presence of the anti-apoptotic antigens, BCl-2 and p53. Cerebellum specimens were obtained from the Laboratory of Animal Pathology, from 1995 to 2009, and the 5-μm thick fragments were stained both with hematoxylin-eosin and Shorr. All samples were diagnosed as positive for CDV genome by reverse transcriptase polymerase chain reaction targeting the nucleocapsid gene. The anti-apoptotic pathways evidenced in this study were BCl-2 and p53 proteins that were intensively detected in cerebellum of CDV positive slides (40-80% of labeled cells/mm2). In addition, the apoptosis markers annexin-V and TUNEL are directly correlated among the same samples (80 and 40% of labeled cells, respectively). This is the first description of p53 and annexin-V expression, characterized as anti-apoptotic and apoptotic proteins, involvement in canine natural cases of CDV infections.

Acute phase proteins in canine lymphoma during antineoplastic chemotherapy

The lymphoma is the main hematopoietic tumor in dogs and it is characterized by the proliferation of cells from lymphoid tissue, histiocytes and its precursors. Animals with lymphoma often show changes in biochemical and hematological parameters such as non-regenerative normochromic normocytic anemia, hemolytic anemia, hypocalcaemia and monoclonal gammopathy. The development of tumor can cause alterations in serum concentrations of acute phase proteins (APPs), consequent of hepatocytes stimulus by cytokines of inflammatory action. This study aimed to quantify and qualify APPs in dogs with lymphoma, at diagnosis time and during the time of chemotherapy sessions. After syneresis, centrifugation and fractioning the serum samples of 10 healthy and 10 dogs with lymphomas, the proteins fractions were separated by polyacrilamide gel electrophoresis (SDS-PAGE) and its concentrations were determined by computer densitometry. Between 18 and 30 proteins were separated by eletrophoresis, with molecular weights ranging from 18 to 245 kDa (kilodaltons). The alpha-1-glicoprotein acid (AGP) and transferrin serum concentration showed significantly higher in dogs with lymphoma, when compared with healthy dogs at diagnosis. The alpha-1-antitripsin (AAT) serum concentrations showed significantly higher in healthy dogs, when compared with dogs with lymphoma at diagnosis. The dogs with lymphoma the albumin did not appear as negative APP. On the other hand, transferrin appeared as positive AAP at diagnosis time and during the chemotherapy sessions. Healthy dogs had AAT serum concentrations significantly higher when compared to dogs with lymphoma at diagnosis. So, in this trial, it is suggested that this protein has been shown as a negative APP in the dogs with lymphoma. These dogs presented significantly higher AGP serum concentrations, in relation to healthy dogs at diagnosis, evidencing this protein APP positive behavior in neoplasm.

Cytoplasmic and nuclear morphometric parameters in cytologic preparations of canine cutaneous mast cell tumors

Thirty fine-needle biopsy (FNB) samples from 28 dogs subjected to surgical resection of cutaneous mast cell tumors (MCTs) were stained with Giemsa. At least 100 neoplastic cells from each cytology slide were evaluated by morphometric analysis. The parameters were: area, perimeter of the cell, cytoplasm, nucleus and circumference factor. MCTs of grade III had a mean cellular area of 231.70 μm2 ± 57.1, and grade II had a mean of 252.30 μm2 ± 55.0. Cellular perimeter was 61.20 ± 7.1 in grade II and 59.1 ± 8.6 in grade III. Cellular parameters were not statistically different between grades (p>.05). Mean nuclear area was 88.90 μm2 ± 19 in grade III and 72.30 μm2 ± 13.9 in grade II, with statistical difference between grades (P =.011). Mean nuclear perimeter was 32.40 ìm ± 3.0 in grade II and 35.70 ìm ± 4.0 in grade III, with statistical difference between grades (P =.018). Mean nuclear circumference factor was 1.0 ± 0.33 in grade II and 1.1 ± 0.28 in grade III, with no statistical difference between grades (P = 0.78). Nuclear-tocytoplasmic ratio in grade II was 0.29 ±.07 and 0.39 ±.08 in grade III, with statistical difference (P =.02). The number of binucleated and multinucleated cells and mitotic figures was significantly increased in grade III MCTs (P <.001). In conclusion, the number of mitotic figures, presence of binucleation and multinucleation, and nuclear-to-cytoplasmic ratio can help to guide a profile of MCT aggressiveness in cytologic preparations.

Canine transmissible venereal tumors: Aspects related to programmed cell death

Canine transmissible venereal tumor (CTVT) is a neoplasm transmitted by the physical transfer of viable tumor cells by direct contact with injured skin and/or mucous tissue. These cells can transpose across histocompatibility barriers into unrelated hosts. This review focuses on the biology of apoptosis and the interaction of proteins involved in this process, as well as p53, p63 and the antiapoptotic protein Bcl-2. As such, this disease offer unique opportunity to study the biology of transplantable tumours and the interaction of proteins involved in apoptosis process and the prognosis of CTVT.

Consensus for the diagnosis, prognosis and treatment of canine mammary tumors

The purpose of this paper is to establish criteria that could guide the diagnosis, prognosis and treatment of canine mammary neoplasias. It was elaborated during the Mammary Pathology Meeting: Diagnosis, Prognosis and Treatment of the Canine Mammary Neoplasm, held on November 6 th and 7 th, 2010 in Belo Horizonte - MG, Brazil, sponsored by the Laboratory of Comparative Pathology - UFMG, with the support of the Brazilian Association of Veterinary Pathology (ABPV) and Brazilian Association of Veterinary Oncology (ABROVET). Academics from several regions of Brazil were present and contributed to this work.

Sorofrequência de infecção pelo vírus da imunodeficiência felina e vírus da leucemia felina em gatos do município de araçatuba, são paulo

Feline immunodeficiency virus and feline leukemia represent important infectious diseases caused by retroviruses. This study aimed to investigate the prevalence of infection by feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) in cats from the municipality of Araçatuba, São Paulo. Blood samples from 302 cats were collected and tested for the presence of antibodies against feline immunodeficiency virus and antigen of feline leukemia virus by ELISA ® Snap- Combo FIV-FeLV (IDEXX Laboratories). The frequency of FIV positivity was 5.63% (17/302) and of FeLV was 0.33% (1/302). Of the 17 cats infected with FIV, nine (52.94%) were symptomatic. There was a prevalence of FIV infection in males (p 0.0316) and cats aged between one and three years (p 0.0324).

Atividade viral do vírus respiratório sincicial bovino (brsv) em bezerros leiteiros

The purpose of the present study was to evaluate the antimicrobial activity of propolis extracts diluted in different solvents against bacteria from Staphylococcus genus. The study was performed in the Immunology and Microbiology Laboratory from Universidade Federal do Vale do São Francisco. The propolis extracts were prepared using brown propolis diluted in different solvents such as chloroform, methanol, ethyl acetate and grain alcohol. In order to determine the antimicrobial potential of extracts, agar well diffusion method was used, with controls for each diluent. After that, Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) methods were used. All tests were performed in triplicate. In the agar well diffusion test, the measurements of the inhibition zone for propolis extract were as follows: grain alcohol and propolis (2.88mm), methanol and propolis (2.41mm), chloroform and propolis (2.40mm) and ethyl acetate and propolis (0.83mm). The MBC of propolis extracts in different solvents were 93.75 μg/mL for grain alcohol, 375 μg/mL for chloroform and methanol and 3,000 μg/ml for ethyl acetate. Statistically significant differences were achieved comparing the inhibition zones of propolis diluted in grain alcohol and ethyl acetate (2.88 and 0.83 mm, respectively). Considering the low cost of therapy and the activity of the propolis against caprine mastitis pathogens, other studies regarding in vivo activity and chemical characterization are necessary, in addition to evaluation of the toxicological aspects of propolis extracts.

Infecção pelo vírus da diarreia viral bovina em vacas gestantes abatidas no Estado de São Paulo-Brasil

The most important pathogens in the bovine livestock nowadays in the virus of the viral diarrhea mainly for triggered clinical manifestations related to the reproductive sphere. The infection in pregnant females, may result in abortions, embryonic resorptions, fetal mummification, birth of weak and malformation of the cattle. Moreover, their birth with persistently infected and immunotolerant virus, which the source of infection and dissemination of their disease. Nowadays, the complexity of the diagnosis and consequently its pathogenesis are tilted in the genotypic differences agent. So, this study aimed to verify the occurrence of the BVDV-1 (SINGER) and BVDV-2 (VS-253) genotypes in cows and their respective fetuses, slaughtered in an abattoir in the State of Sao Paulo. Through blood serum, using virus neutralization technique. All in all, 52,51% (115/219) of the cows which were tested reacted, but no fetus (0/219) reacted, to its virus neutralization. Through this cross-examination we observed that 42% (92/219) of cows reacted for both BVDV-1 and BVDV-2. Furthermore 4,10% (9/219) of them reacted only to the genotype BVDV-1 and 6,39% (14/219) responded only to the genotype 2 of BVDV. Therefore it was noticed that both strains are widespread in the regions studied, which justifies the use of different antigens to avoid false-negative diagnosis. Finally antibodies showed no fetus or fetal abnormalities, it is already developed and can be considered immunocompetent, independent child born to a reagent.

Falhas reprodutivas associadas com a presença de do vírus da artrite-encefalite caprina, toxoplasma gondii e neospora caninum em caprinos no estado de São Paulo, Brasil

This study aimed at assessing the occurrence of antibodies against the caprine arthritis-encephalitis virus (CAEV), Toxoplasma gondii and Neospora caninum, as well as the associations between the presence of antibodies and the occurrence of reproductive failures in goats. Serum samples were collected from 923 goats of both sexes, over 3 months of age, from 17 dairy farms located in different municipalities of São Paulo State, Brazil. Infections by T. gondii, N. caninum and CAEV were evaluated by indirect methods of diagnosis based on indirect fluorescence antibody test (IFAT), Neospora agglutination test (NAT), and agar gel immunodiffusion (AGID), respectively. A survey was conducted on the farms to obtain information about reproduction dates (abortions, stillbirths and births of weak and premature kids) and zoosanitary management. Antibodies against CAEV, T. gondii and N. caninum was found in 37.81%, 23.62% and 17.23% respectively. There was no significant association between the presence of anti-CAEV antibodies and CAEV/T. gondii or CAEV/N. caninum co-infection, suggesting that CAEV does not predispose goats to infection by these agents. However, when CAEV/T. gondii (p<0.01) or CAEV/N. caninum (p<0.001) co-infection was present, the occurrence of reproductive failures was significantly higher what could indicate that CAEV-induced immunosuppression may predispose goats to develop the clinical symptoms of toxoplasmosis and neosporosis increasing the risks of the reproductive failures.

Caracterização genética do vírus da diarreia bovina detectada em bovinos do Sul de Minas Gerais, Brasil, persistentemente infectados

Isolates of bovine viral diarrhoea virus (BVDV) detected in serum samples of two persistently infected animals (PI) identified in a herd located in the southern state of Minas Gerais, Brazil, underwent genetic characterization trough partial nucleotide sequencing and analysis of the 5' Untranslated Region (5'UTR) of the viral genome. The isolates were characterized as belonging to genotype BVDV-1, subgenotype BVDV-1b. The results of this study suggest BVDV-1b as an agent of importance in the occurrence of bovine viral diarrhoea (BVD) in the herds of the region. Moreover, the genotypic characterization of isolates of BVDV helps to better understand the epidemiology of the disease, as the genetic variability of BVDV interferes in the serological tests and has implications for the use of vaccines, whose majority is produced only with reference strains of BVDV. Therefore, the investigation on the genetic diversity of BVDV existing in Brazil is required for the improvement of the disease prevention and control measures.

Foetal echoencephalography and Doppler ultrasonography of the middle cerebral artery in canine foetuses

Objectives: This study aimed to evaluate the characteristics of the brain and vascular indicesof the middle cerebral artery of canine foetuses. Methods: Twenty-five bitches were selected. Tissue development, echogenicity, echotexture and brain echobiometric data were studied, and the major structures were identified between the 5th and 8th gestational weeks. The area and volume of the brain mass (BMA and BMV), cranial area and volume (AC and VC), brain mass index (BMI) and brain volume index (BVI) were determined. A single ultrasound examination was performed during each studied week (6th, 7th and 8th). Doppler ultrasonography was performed to assess the maximum and minimum velocity, resistance and pulsatility index of middle cerebral artery of the foetuses. Results: Echoencephalography was performed to evaluate the morphological characteristics of the central nervous system. Cerebral echobiometry indicated an increase in area and volume of the hemispheres and cranium (P<0·001) but no changes in BMI or BVI over the gestational period studied. Doppler ultrasonography identified increases in peak systolic velocity (P=0·0188) and end diastolic velocity (P=0·0274) and decreases in resistance index (P=0·0002) and pulsatility index (P<0·001). Clinical Significance: Echoencephalography and spectral Doppler ultrasonography of the middle cerebral artery in canine foetuses might be a useful technique for prenatal care. © 2013 British Small Animal Veterinary Association.