Interferon-γ regulates cell malignant growth via the c-Abl/HDAC2 signaling pathway in mammary epithelial cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Interferon-γ (IFN-γ) has been used to control cancers in clinical treatment. However, an increasing number of reports have suggested that in some cases effectiveness declines after a long treatment period, the reason being unclear. We have reported previously that long-term IFN-γ treatment induces malignant transformation of healthy lactating bovine mammary epithelial cells (BMECs) in vitro. In this study, we investigated the mechanisms underlying the malignant proliferation of BMECs under IFN-γ treatment. The primary BMECs used in this study were stimulated by IFN-γ (10 ng/mL) for a long term to promote malignancy. We observed that IFN-γ could promote malignant cell proliferation, increase the expression of cyclin D1/cyclin-dependent kinase 4 (CDK4), decrease the expression of p21, and upregulate the expression of cellular-abelsongene (c-Abl) and histone deacetylase 2 (HDAC2). The HDAC2 inhibitor, valproate (VPA) and the c-Abl inhibitor, imatinib, lowered the expression level of cyclin D1/CDK4, and increased the expression level of p21, leading to an inhibitory effect on IFN-γ-induced malignant cell growth. When c-Abl was downregulated, the HDAC2 level was also decreased by promoted proteasome degradation. These data suggest that IFN-γ promotes the growth of malignant BMECs through the c-Abl/HDAC2 signaling pathway. Our findings suggest that long-term application of IFN-γ may be closely associated with the promotion of cell growth and even the carcinogenesis of breast cancer.

Somatic cell count and mastitis pathogen detection in composite and single or duplicate quarter milk samples / Contagem de células somáticas e detecção de patógenos causadores de mastite em amostras de leite compostas e individuais simples ou em duplicata

The most acceptable criteria for diagnosing bovine intramammary infections include results of bacteriological culture and measures of inflammation. Therefore, information on the diagnostic characteristics of the procedures used to identify infected quarters is required. Thus, this study was designed to evaluate a set of criteria to classify the infectious status of an udder at the quarter (single and duplicate milk samples) and cow (composite milk sample) levels, and to compare the infectious status with somatic cell counts (SCCs) of the samples. Here, the SCC thresholds determined by receiver operating characteristic curve analysis had a higher Youden index using mammary quarter duplicate milk samples as the gold standard for testing compared with single quarter and composite milk samples, especially for samples for which at least one of the duplicates was microbiologically positive, regardless of the mastitis pathogen isolated. The kappa coefficient for bacteriological results of the single quarter milk samples (single S1 and S2) was 0.85±0.019, indicating that single quarter milk sampling can be useful in mastitis control programs. Therefore, the use of composite milk samples to detect mastitis pathogens may be limited to the detection of major pathogens, given their predictive values. Thus, our findings suggest that the milk SCCs and microbiological examinations, although regarded as the most reliable indicators of ongoing mastitis, should be used in an integrated manner in mastitis control programs. Furthermore, the accuracy of single, duplicate and composite microbiological analyses to diagnosis mastitis should be considered for its implications in mastitis control strategies.(AU)

Os critérios mais aceitáveis para o diagnóstico das infecções intramamárias em bovinos incluem tanto os resultados da cultura bacteriológica e dos indicadores de inflamação. Portanto, a informação sobre os procedimentos mais adequados a serem utilizados para identificação dos quartos infectados é necessária. Assim, o objetivo do presente estudo foi avaliar um conjunto de critérios para identificação da infecção intramamária em bovinos pelo exame microbiológico (amostras individuais de leite simples ou em duplicata, e amostras de leite compostas), e comparar o isolamento do patógeno nas amostras de leite coletadas por distintos critérios com a contagem de células somáticas (CCS). Os valores de corte da CCS determinados pela curva de característica de operação do receptor demonstraram que a coleta de amostras de leite em duplicata apresentou o maior valor do índice de Youden, especialmente quando considerou-se o quarto mamário infectado se pelo menos uma das amostras de leite da duplicata apresentou resultado bacteriológico positivo independentemente do patógeno isolado. O coeficiente kappa dos resultados do exame microbiológico das amostras de leite individuais (amostra simples S1 e S2) foi de 0,85±0,019, indicando que a coleta de amostras de leite individual, ou seja, a coleta de uma amostra de leite por quarto mamário, pode ser utilizada nos programas de controle de mastite. Por outro lado, a coleta de amostras de leite compostas para detectar patógenos causadores de mastite deve ser limitada à detecção dos patógenos principais, considerando os valores preditivos encontrados no presente estudo. Portanto, os resultados do presente estudo indicam que a CCS e o exame microbiológico do leite, embora considerados como os critérios mais aceitos para o diagnóstico da mastite, devem ser utilizados de forma integrada em programas de controle de mastite. Além disto, os critérios de coleta de amostras de leite para o diagnóstico da mastite pelo exame microbiológico e seus valores preditivos devem ser considerados nos programas de controle de mastite.(AU)

Somatic cell counts as an indicator of mammary infection in periparturient cows / Contagem de células somáticas como indicador de infecção mamária em vacas periparturientes

The somatic cell count (SCC) is a screening test for the evaluation of intramammary infection; however, changes in mammary secretion during colostrogenesis can promote a physiological increase in the SCC, potentially reducing its reliability in the diagnosis of mastitis. Therefore, the aim of this study was to evaluate bovine colostrum SCC as an indicative parameter of breast infection in Holstein periparturient cows. A total of 80 samples were harvested from the first milking colostrum of 20 cows and were subjected to manual SCC and bacteriological examination. Bacterial growth was present in 36.62% of the crops; coagulase-negative Staphylococcus (CNS) was the predominant microorganism (76.92%). The median SCC in infected cows (1.8 × 106 cells/mL) was significantly higher than in uninfected cows (0.9 × 106 cells/mL) (p = 0.0451). The sensitivity (10015%), specificity (1002.2%), and false positive (1002.2%) of the SCC decreased gradually when thresholds of 0.210.0 × 106 cells/mL were adopted. In contrast, the proportion of false negatives (084.6%) revealed an opposite trend. The threshold of greatest concordance between SCC and bacteriological examination was 10.0 × 106 cells/mL; however, the sensitivity rates (15.4%), specificity (2.2%), and false positive (2.2%) were very low. Based on these results, we conclude that SCCs increased prior to the infectious processes of the mammary gland, particularly in the CNS group. However, physiological changes caused by colostrogenesis resulted in poor concordance between the SCC and bacteriological examination of the colostrum...

A contagem de células somáticas (CCS) é um teste de triagem para avaliação de infecção intramamária, porém, as alterações na composição do colostro observadas durante a colostrogênese podem promover o aumento fisiológico da CCS e inviabilizar o uso desta prova para diagnóstico da mastite. Assim, o objetivo desta pesquisa foi avaliar a CCS do colostro como parâmetro indicativo de infecção mamária em vacas Holandesas periparturientes. Para tanto, foram colhidas 80 amostras de colostro de primeira ordenha, provenientes de 20 vacas, para a CCS manual e exame bacteriológico (EB). Observou-se crescimento bacteriano em 36,62% dos cultivos, com predomínio de Staphylococcus coagulase-negativa (SCN) (76,92%). O valor mediano da CCS de vacas infectadas (1,8 x106 células/mL) foi maior do que o obtido para as vacas sadias (0,9 x106 células/mL) (P=0,0451). A sensibilidade (100 a 15%), especificidade (100 a 2,2%) e falso-positivo (100 a 2,2%) diminuíram gradativamente quando os limiares de 0,2 a 10,0 x106 células/mL foram adotados. Em contrapartida, a proporção de falso-negativo (0 a 84,6%) apresentou perfil inverso. O limiar de maior concordância entre a CCS e EB foi de 10,0 x106 células/mL, porém os índices de sensibilidade (15,4%), especificidade (2,2%) e falso-positivo (2,2%) foram muito baixos. Com base nos resultados encontrados, conclui-se que a CCS apresentou elevação diante dos processos infecciosos da glândula mamária (GM), causados especialmente por bactérias do grupo SCN. No entanto, as alterações fisiológicas decorrentes da colostrogênese resultaram em baixa concordância entre a CCS e o exame bacteriológico do colostro...

Dispositivo fotobiomodulador para prevenção e tratamento de hiperqueratose de teto em vacas leiteiras / Photobiomodulation device for prevention and treatment of teat hyperkeratosis in dairy cows

As mastites estão entre as principais causas de prejuízo para produtores de leite. Em casos graves de hiperqueratose, o canal do teto pode se tornar uma barreira mais fácil para que as bactérias penetrem na glândula mamária. Os objetivos deste estudo foram avaliar um dispositivo fotobiomodulador de LED para tratamento e prevenção de hiperqueratose de teto e prevenção da mastite subclínica em um rebanho de leite com alta prevalência de hiperqueratose (35,3 por cento de casos graves). Foram utilizadas 60 primíparas para o experimento de prevenção e 30 vacas com hiperqueratose para o experimento terapêutico. Em ambos os experimentos, metade dos animais foram tratados com o dispositivo fotobiomodulador três vezes por semana, durante 6 semanas. Os outros animais foram os controles. Imagens fotográficas digitalizadas foram realizadas na avaliação inicial e, semanalmente, por 6 semanas consecutivas. Nas primíparas, novas avaliações foram realizadas entre 6 e 7 meses de lactação. Para avaliação da mastite subclínica, contagem de células somáticas (CCS) foram feitas mensalmente. No experimento preventivo, o diâmetro externo das lesões permaneceu constante nos tetos do grupo tratado, enquanto houve aumento no grupo controle. No experimento terapêutico não foram observadas diferenças estatísticas entre as variáveis de hiperqueratose. Contudo, o grupo tratado apresentou menor incidência de mastites subclínicas (CCS < 250 células/mL) por lactação do que o grupo controle (P<0,05). Em conclusão, o tratamento não foi efetivo em prevenir o desenvolvimento ou reduzir lesões instaladas de hiperqueratose de teto. Contudo, o uso protótipo se mostrou útil e promissor como adjuvante na prevenção do aumento de tamanho das lesões de hiperqueratose de teto em primíparas e como forma de reduzir incidência de mastite subclínicas em vacas leiteiras já acometidas.

Mastitis consistis one of the main problems of milk production, mainly due to the production losses and the rising cost of milk. In case of severe hyperkeratosis, the teat canal can become an easier barrier for the bacteria to penetrate. The objeSctive of this study was to assess a phototherapy device construct with LED light for prevention and treatment of teat hyperkeratosis in a dairy cattle herd with high prevalence (35.3 percent of severe cases). 60 primiparous cows were used in the preventive experiment and 30 cows with hyperkeratosis were used in the therapeutic experiment. In both experiments, half of the cows started to be treated in early lactation using the phototherapy device three times per week for 6 weeks. The other cows were the controls. Hyperkeratosis was assessed by scoring and by morphometric analyses of teat images that were taken at baseline and then weekly for 6 consecutive weeks. In the preventive experiment, more images were taken later, between 6 and 7 months of the first lactation. Somatic cell counts (SCC) were performed monthly. In the preventive experiment, the outer diameter of the teat lesions remained constant in the treated group, whereas cows of the control group showed a significant increase at the end of lactation. In the therapeutic groups, no statistical differences for teat hyperkeratosis variables were seen between groups. However, the incidence of subclinical mastitis (SCC > 250 cells/mL) during lactation was lower in the treated group (P<0.05). In conclusion the phototherapy protocol did not prevent the development in primiparous or ameliorate previous hyperkeratosis lesions in cows. However, the prototype was considered useful as an adjunct in preventing the increase in the size of teat hyperkeratosis lesions and also as a way to reduce subclinical mastitis incidence in affected dairy cows.

Histomorfometria da mama de ratas tratadas com estrogênio e/ou progestagênio / Breast histomorphometry of rats treated with estrogen and/or progestogen

OBJETIVO: Avaliar as alterações histomorfométricas nas mamas de ratas tratadas com estrogênio e/ou progestagênio por curto período de tempo. MÉTODOS: Foram divididas em quatro grupos 40 ratas ooforectomizadas: GC-recebeu veículo; GE-recebeu benzoato de estradiol (37,6 µg/animal); GP-recebeu acetato de medroxiprogesterona (11,28 mg/animal) e, GEP-recebeu benzoato de estradiol (37,6 µg/animal) e acetato de medroxiprogesterona (11,28 mg/animal). No grupo GE, o estradiol foi administrado durante sete dias, por via subcutânea. Já no grupo EP o estradiol foi administrado nos primeiros sete dias e o progestagênio por mais 23 dias, por via subcutânea. Vinte e quatro horas após a última administração dos hormônios, os animais foram anestesiados e o primeiro par de mamas inguinais removido, imerso em formaldeído a 10 por cento e processado para inclusão em parafina, sendo os cortes corados pela Hematoxilina-Eosina. Foram avaliadas a morfologia e a área ocupada pelo parênquima mamário, sendo os dados submetidos à análise de variância complementado pelo teste de Kruskal-Wallis (p < 0,05). RESULTADOS: As mamas no grupo-controle apresentaram-se atrofiadas, sendo que, nos animais dos grupos GE e GEP, nota-se a presença de alvéolos típicos contendo secreção no seu interior, já nos animais tratados somente com progestagênio (GP) notam-se alvéolos formados por células volumosas que ocupam praticamente todo o lúmen alveolar. A morfometria mostrou haver maior área de parênquima mamário nos animais tratados com hormônios (GE = GP > GEP > GC; p < 0,05) CONCLUSÃO: O estradiol e o progestagênio apresentaram efeito proliferativo no parênquima mamário. No entanto, a administração prévia de estradiol modifica a ação do progestagênio no tecido mamário da rata.

OBJECTIVE: To evaluate the breast histomorphometric changes in rats treated with estrogen and/or progestogen for a short period of time. METHODS: Forty oophorectomized rats were divided into four groups: GC, vehicle; GE, treated with estradiol benzoate (37.6 mg/animal); GP, treated with medroxyprogesterone acetate (11.2 mg/animal) and GEP, treated with estradiol benzoate (37.6 mg/animal) plus medroxyprogesterone acetate (11.28 mg/animal). In GE group, estradiol was administered subcutaneously for seven days; in GEP group, estradiol was administered once in a day for the first seven days and the progestogen over the next 23 days both subcutaneously. Twenty-four hours after the last hormone administration, the animals were killed upon deep anesthesia and the first inguinal breasts were removed, fixed in 10 percent formaldehyde and processed to be included in paraffin, with the sections being stained by hematoxylin-eosin. Morphology and the area occupied by mammary parenchyma were assessed, with the data undergoing analysis of variance followed by the Kruskal-Wallis test (p < 0.05). RESULTS: The control group breasts were found atrophic and, in GE and GEP group animals, typical alveoli with secretion inside are present; in progestogen-treated animals (GP), alveoli formed by large cells occupying almost the entire alveolar lumen are noted. Morphometric analysis showed a larger mammary parenchyma area in hormone-treated animals (GE = GP > GEP > GC; p < 0.05). CONCLUSION: Estradiol and progestogen had a proliferative effect on mammary parenchyma. However, prior estradiol administration changes the progestogen action on rat mammary tissue.

Variaciones nucleolares en la transformación celular del epitelio mamario / Nucleolar variations in the cell transformation of mammary epithelium

Antecedentes: En la medida que las células pertenecientes a la línea HC11 son transfectadas con el onco-gén ras, asumen distintas propiedades resultando en tipos celulares transformados, modificando tanto sus componentes como sus funciones celulares, los cuales pueden ser cuantificadas mediante técnicas morfométricas. Objetivo: Evidenciar en términos cuantitativos y morfológicos las variaciones experimentadas por los nucleolos pertenecientes a células mamarias de la línea HC11 con el decorrer del mecanismo de transformación celular. Método: Se estudió a nivel de la microscopia electrónica de transmisión los tipos celulares en proceso de transformación (Q6 GM), en comparación con células francamente neoplásicas (Q6 IM), cuantificando variaciones de los nucleolos y su relación con estructuras involucradas en síntesis proteica. Resultados: Se evidencian diferencias estadísticamente significativas en el área, volumen y longitud entre los nucleolos pertenecientes a estos tipos celulares. Conclusión: Las células del epitelio mamario en proceso neoplásico presentan un notable aumento de sus nucleolos y sus ribonucleoproteínas, constituyentes que generarán básicamente ribosomas libres, que sintetizarán proteínas para ser utilizadas en el decorrer de las mitosis sucesivas y desreguladas.

Background: As cells belonging to the HC11 line become transfected with the ras oncogene, they assume different properties resulting in transformed cell types, with modified cell components and functions. These may be quantified by morphometric techniques. Objective: To provide quantitative and morphological evidence of the variations occurring in the nucleoles of HC11 line mammary cells as the cell transformation mechanism takes its course. Method: Transmission electron microscopy was used to study cell types in the transformation phase (Q6 GM) as compared with frankly neoplastic cells (Q6 IM), quantifying the variations between the nucleoles and their relation to structures involved in protein synthesis. Results: Statistically significant differences were found between the nucleoles belonging to these cell types with respect to area, volume and length. Conclusion: The nucleoles of mammary epithelial cells in the process of neoplasia, present a notable increase, and their constituent ribonucleoproteins will basically generate free ribosomes, synthesising proteins available for use in successive, unregulated mitosis.

Condrosarcoma mesenquimal sinonasal grado tres y carcinosarcoma de glándula Mamaria de un canino: estudios clínico e histopatológico / Sinunasal mesenchymal chondrosarcoma grade 3 and mammary gland carcinosarcoma of canine: clinical and histopathology study

A la Clínica Veterinaria de la Universidad de los Llanos, llegó un canino hembra de 9 años de edad raza Chow Chow, cuyo motivo de consulta fue epistaxis bilateral 4 meses atrás y una masa en la región nasal. Según lo reportado por el propietario, la masa había comenzado a salir 1 mes atrás del día de la consulta. Al examen clínico presentaba una masa subdérmica en la región nasal superior de 2 cm de diámetro. Se realizó una citología de la misma y se encontraron células inflamatorias sin presencia de células neoplásicas. El paciente volvió al mes y la masa con un diámetro de 5cm, estaba ubicada en la región nasal superior y región zigomática. El can presentaba disnea, anorexia, deshidratación del 6 por ciento, secreción mucopurulenta nasal y ocular, midriasis bilateral, reflejo corneal bilateral negativo, aumento de la presión infraocular del ojo derecho. En la glándula mamaria inguinal derecha presentaba una masa de 2cm de diámetro dura y móvil. Se realizó una citología por aspirado con aguja fina, de distintas zonas de las masas, tanto de la nariz, como de la glándula mamaria. Se valoró la citología permitiéndose definir el origen tumoral del proceso. Se realizó biopsia por incisión de la región sinonasal, diagnosticándose tumor mesenquimal maligno grado tres condrosarcoma sinonasal, conformado por células cartilaginosas pleomórficas con anisocariosis. Se realizaron radiografías de la región nasal y del pulmón, encontrándose una zona radiopaca a nivel del seno maxilar con osteolisis del hueso, se evidenciaba la masa de aproximadamente 4 cm de ancho por 2 cm de largo en el lado derecho. En el pulmón no se evidenció metástasis. Después del diagnóstico de histopatología se decide realizar la eutanasia, encontrándose histopatológicamente metástasis en pulmón, confirmación del condrosarcoma mesenquimal y un carcinosarcoma de glándula mamaria.

To the Veterinary Clinic of the Universidad de los Llanos, come a 9 years old Chow Chow female canine, whose consultation reason was a bilateral epistaxis occurred 4 months ago, sudden loss of the vision and a mass in the nasal region of 2cm of diameter approximately, according to the report of the owner, the mass showed up 1 month the day of the consultation. To the clinical exam they were the following abnormal discoveries. Hirsute hair, opaque, presented a subdermic mass approximately in the nasal region superior of 2 cm. diameter that grew to 5 cm. diameter after 1 month of the consultation, located in the nasal region superior and zigomatic region, of hard consistency, the animal had dysnea, bilateral mydriasis, the bilateral corneal reflection was negative, increase of the infraocular pressure of the right eye, the realized test of obstacles in the clinic was negative, that which showed us a blindness. Besides a complete clinical exploration and taking of the advantages of the cytology, she was carried out an aspired with fine needle of different areas of the masses so much neoplastic of the nose and of the region of the mammary gland, being obtained two types of samples. The cytology was evaluated to define origin of the tumors of the process. She was carried out biopsy for incision of the sinunosal region being diagnosed wicked tumor mesenchymal Chondrosarcoma sinunasal that commits bone. After having carried out the autopsy they took samples for histopathology. Being diagnosed mesenchymal chondrosarcoma sinunasal grade 3, with metastasis to lung, and a carcinosarcoma of mammary gland.

Proliferação, apoptose e histomorfometria da glândula mamária de ratas tratadas com tiroxina na lactação e ao desmame e desenvolvimento dos filhotes / Proliferation, apoptosis and mammary gland histomorphometry of thyroxine-treated rats on lactation and after weaning and development of offspring

O objetivo deste estudo foi avaliar a histomorfometria e a taxa de proliferação e apoptose da glândula mamária de ratas tratadas com tiroxina pela imuno-expressão de CDC-47 e caspase-3, respectivamente. Também foi avaliado o desenvolvimento dos filhotes de ratas tratadas com tiroxina. Foram utilizadas 36 ratas distribuídas em dois grupos, tratado com tiroxina e controle. Após 60 dias de tratamento com tiroxina, as ratas foram acasaladas. Seis animais/grupo foram sacrificados no 2° e 21° dias de lactação e no 5° dia após o desmame. Houve diferença significativa entre grupos apenas no quinto dia após o desmame. O tratamento com tiroxina aumentou a taxa de apoptose caracterizada pela maior expressão de caspase-3 nas células do epitélio mamário. As mães tratadas com tiroxina apresentaram comportamento alterado, mas não houve diferença significativa no que se refere aos cuidados com o filhote quanto a higienização e aquecimento. Levando-se em consideração o sexo e o tamanho da ninhada, os filhotes das ratas tratadas com tiroxina e controle não apresentaram diferença significativa de peso ao desmame. Conclui-se que a administração de baixas doses de tiroxina aumenta a taxa de apoptose, caracterizada pelo aumento da expressão de caspase-3 no epitélio mamário cinco dias após o desmame, mas não altera a taxa de proliferação celular e o comportamento materno.

The purpose of this study was to evaluate mammary gland histomorphometry and proliferation rate and apoptosis of thyroxine-treated rats by CDC-47 and caspase-3 immunoexpression. The development of thyroxine-treated rats offspring was also evaluated. Thirty-six female rats were used, distributed in two groups, treated and non-treated with thyroxine. After 60 days of treatment, with thyroxine, rats were mated. Six animals/group were sacrificed on the 2nd and 21st days of lactation and on the 5th day after weaning. A significant difference was observed between groups only on the 5th day after weaning. Thyroxine treatment increased apoptosis rate, which was characterized by a higher caspase-3 expression in mammary epithelial cells. Thyroxine-treated mothers presented changed behavior, but there was no significant difference regarding taking care of offspring, as for cleaning offspring and keeping them warm. Taking into account sex and size of offspring, those from control and thyroxine-treated mothers presented no significant difference of weight and weaning. In conclusion, administering low doses of thyroxine increases apoptosis rate, which is characterized by the increased caspase-3 immunoexpression in mammary epithelial cells 5 days after weaning. But does not affect proliferation rate and development of thyroxine-treated rats offspring.

Mammary fibroadenoma in a lamb

A fibroadenoma was diagnosed in the left udder of a 3-month-old female Chios lamb. No recurrence was observed after surgery. Grossly, the tumor had a whitishgray lobular appearance, and the lobules were interlaced with thin septa. Microscopically, the tumor was composed of proliferating fibroepithelial tissue, including differentiated ducts lined by whorls and interlacing bundles of abundant loose fibrovascular stroma. Immunohistochemistry revealed the ductal epithelium to be positive for pancytokeratin (AE1/AE3) and loose fibrovascular stroma was positive for vimentin and basal cells covering the ductal epithelium of alpha-smooth-muscle actin. Immunostaining for the estrogen and progesterone receptors was negative. A diagnosis of mammary fibroadenoma was made based on the histological and immunohistochemical findings.

Proteinograma do leite de vacas lactantes submetidas à retenção láctea / Milk proteinogram of the cows submitted to milk retention

Doenças, intensidade na criação e produção, bem como o manejo inadequado foram fatores considerados estressantes para vacas leiteiras, determinando a retenção de leite. Quatro vacas adultas, sadias e em plena lactação e sem antecedentes de mamites e/ ou tratamento intra-mamário foram submetidas experimentalmente a 10% de retenção de leite, das quais colheram-se amostras de leite nos seguintes momentos (Tempos): antes da retenção, 12, 24, 36, 48, 60 horas durante a retenção e, 168 e 180 horas após o início do procedimento, ou seja, 108 e 120 horas após cessada a retenção. As amostras de leite foram, previamente, submetidas a exames físico-químico e microbiológico. O soro lácteo era obtido por coagulação do leite com renina e o proteinograma determinado por biureto e fracionamento por eletroforese em gel de poliacrilamida. Observou-se um gradativo e significativo aumento de algumas frações do soro lácteo: albumina e imunoglobulina sérica bovina; lactoferrina; alfa1-antitripsina; ß-lactoglobulina e; alfa-lactoalbumina. Ao final da experimentação os valores das frações protéicas, retomaram aos iniciais, momento anterior ao início da retenção láctea.

Diseases, breeding and production intensity, as well as inadequate handling were factors considered as stressful to rnilk cows, determining milk retention. Four healthy adult cows and in full milking period and without any prior mamits and/or intra-mammary treatment were experimentally submitted to 10% milk retention of which milk samples were collected at the following times: before milk retention, 12,24,36,48,60 hours during retention and 168 and 180 hours after the initial procedure, or being, 108 and 120 hours after retention ceased. The milk samples were previously submitted to physic-chemical and microbiological exams. The whey was obtained by milk coagulation with renin and the proteinogram determined by biuret method and fractionizing by polyacrymalide gel electrophoresis. A gradual and significant increase of some fractions of the whey was observed: serum albumin and immunoglobulin bovine, lactoferrin; alpha 1-antitrypsin; ß-lactoglobulin and; alpha-lactoalbumin. At the end of the experiment, the proteic fractions returned to their initial state, the moment previous to the initial milk retention.

Corynebacterium bovis e os padrões de contagem de células somáticas no Brasil / The corynebacterium bovis and the somatic cell counting standards in Brazil / El Corynebacterium bovis y los paradrones celulares somáticos en brasil

Foram estudadas 60 amostras de leite de glândulas mamárias bovinas naturalmente infectadas pelo Corynebacterium bovis em estado puro, exuberante e que apresentavam pelo menos uma glândula contralateral com cultivo microbiológico negativo, provenientes de cinco propriedades leiteiras localizadas no interior do estado de São Paulo. Essas foram cultivadas em ágar sangue ovino e em agar MacConkey e submetidas à contagem eletrônica de células somáticas. Obtiveram-se medianas de contagens de células somáticas (CCS), 262 x 10³ céls/mL e 806 x 10³ céls/mL de leite, para as amostras negativas e positivas ao Corynebacterium bovis respectivamente, sendo que, em alguns casos positivos, a CCS superou 10.000.000 céls/mL de leite. Os dados obtidos no presente estudo sugerem que o Corynebacterium bovis é responsável por alterações celulares significativas no leite, podendo acarretar prejuízos aos produtores e à indústria de laticínios, na medida que se observam os padrões celulares stabelecidos pelo Ministério da Agricultura, Pecuária e Abastecimento em vigor desde setembro de 2002.

ABSTRACT: Sixty milk samples from bovine mammary glands naturally infected by Corynebacterium bovis in pure state, which presented at least one contralateral gland with negative microbiological culture from fi ve dairy farms in the State of Sao Paulo were studied. These samples were cultivated in MacConkey agar and submitted to an electronic Somatic Cell Counting (SCC). The medians of SCC results were 262 x 103 cells/mL and 806 x 103 cells/mL from negative and positive milk samples to C. bovis respectively. In some positive samples to the agent, it was observed SCC over 10.000.000 cells/mL of milk suggesting that C. bovis may be responsible for signifi cant cell alterations in the milk, which might cause serious economic damages to the producers and to the milk industry, according to the cell standards established by Agriculture Ministry, Brazilian law from September of 2002

RESUMEN: Fueron estudiadas sesenta muestras de leche de glándulas mamarias bovinas naturalmente infectadas por el Corynebacterium bovis en cultivo puro, provenientes de cinco haciendas lecheras localizadas en el interior del estado de São Paulo. Las muestras fueron cultivadas en agar sangre ovino y en agar MacConkey, y sometidas a cómputo electrónico de células somáticas. Se obtuvo medianas de los cómputos de células somáticas, 262 x 103 céls/mL y 806 x 103 céls/mL en la leche, para las muestras negativas y positivas al Corynebacterium bovis respectivamente, siendo que en algunos casos positivos, la CCS superó 10.000.000 céls/mL de leche. Los datos obtenidos en el presente estudio sugieren que el Corynebacterium bovis es responsable por alteraciones celulares signifi cativas en la leche, pudiendo acarrear perjuicios a los productores y a la manufactura de lácteos, en la medida que se observan los moldes celulares establecidos por el Ministerio de la Agricultura, Pecuaria y Abastecimiento en vigor desde setiembre de 2002

Morphological changes induced by testosterone in the mammary glands of female Wistar rats

Increased levels of androgens in postmenopausal women are considered to be a risk factor for breast cancer. Testosterone, alone or in combination with estrogen, induces epithelial dysplasia and mammary tumors in Noble rats. Since this model of hormone-induced neoplasia has not been reported in other rat strains, we studied the effect of testosterone on the mammary gland morphology of female Wistar rats. Sixty adult, non-castrated, female Wistar rats were implanted in the dorsum midline with a silicone tube containing 50 mg testosterone (testosterone propionate in 30 animals and non-esterified testosterone in the remaining 30 animals) and 20 additional animals were implanted with empty tubes and used as control. Five animals per group were killed 30, 60, 90, 120, 150, and 180 days after implantation, and the mammary glands were dissected, fixed and embedded in paraffin. Histological sections were then stained with hematoxylin and eosin and picrosyrius red for collagen visualization. Morphological and morphometric analysis demonstrated ductal proliferation and acinotubular differentiation with secretory activity in all treated animals, peaking at 90 days of androgen exposure. After 90 days the proliferation of acinar epithelial cells was evident, but there was a progressive reduction of secretory differentiation and an increase in intralobular collagen fibers. There was no morphological evidence of dysplastic changes or other pre-neoplastic lesions. Testosterone treatment applied to adult, non-castrated female Wistar rats induced a mammary gland hyperplasia resembling the lactating differentiation, with progressive reduction in secretory differentiation.

Lead poisoning during development: effect on some antioxidant enzymes, residual level and histopathological changes in rats

Pregnant, pregnant and lactating and/or lactating groups of female rats were exposed to lead in the form of lead acetate in the drinking water at a dose level of 200 mg/L for periods of gestation only [GI], gestation and lactation [GII] and lactation only [GIII]. Blood samples, liver, brain and mammary gland were taken from the treated dams as well as brain tissue samples from their babies at the end of each exposure period. Lipid peroxidation and the activity of both reduced glutathione [GSH] and catalase enzymes were evaluated in liver and brain tissues of dams as well as in the brain tissue of babies. Determination of the residual level of lead was carried out in serum, liver and brain tissue of the dams. Histopathological examination was performed in the liver, brain and mammary gland tissues of treated dams. Our results revealed that exposure to lead induced oxidative tissue damage, which was reflected by significant increase in the level of malondialdehyde [MDA] and decreased activities of GSH and CAT in tissues of dams and babies. Residual concentrations of lead were significantly increased in brain, serum and liver in comparison to control group. The histopathological examination revealed marked histological alterations in the liver, brain and mammary gland of treated dams. This investigation indicated that the effect of lead was more obvious in gestation and lactation period [GII] than the other two periods [gestation only [GI] and lactation only [GIII]]. Also, it denoted that the brain is the primary target organ of lead poisoning

El anticuerpo monoclonal Ki-67 como elemento de valor diagnóstico y pronóstico en neoplasias mamarias caninas / Monoclonal antibody KI-67 as a tool in diagnosis and prognosis in canine mammary tumours

Para muchos el problema del cáncer, es solo un fenómeno humano, sin embargo, éste también afecta a los animales domésticos. Al igual que el Hombre, los animales de compañía (perros y gatos) viven cada vez más, debido a una mejor calidad de vida, una adecuada medicina preventiva y programas de vacunación. Una vida más larga implica un aumento en las posibilidades de desarrollar cáncer. En los últimos años, las técnicas inmunohistoquímicas han contribuido a mejorar el diagnóstico y pronóstico en los casos de neoplasias. Entre tales técnicas destaca el anticuerpo monoclonal Ki-67, el cual detecta un antígeno nuclear que se expresa en las células que entran al ciclo celular, proporcionando una medida directa de la fracción de crecimiento del tejido. Para determinar la utilidad del anticuerpo Ki-67 en neoplasias mamarias de caninos, se aplicó la técnica inmunohistoquímica a 31 muestras de estas neoplasias: 14 neoplasias benignas y 17 malignas. Las neoplasias benignas presentaron un índice de proliferación (IP) promedio de 12.2 por ciento. Las neoplasias malignas presentaron un IP promedio de 19.3 por ciento. Los resultados obtenidos indicaron que los tumores benignos presentaron un índice de proliferación inferior al 15 por ciento y los tumores malignos presentaron IP sobre el 15 por ciento. El anticuerpo Ki-67 puede constituir una importante herramienta diagnóstica y pronostica para la Medicina Veterinaria.

Prototheca zopffi como agente de mastite bovina: clínica e terapêutica / Prototheca zopffi as agent of bovine mastitis: clinics and therapeutics

It was described for the first time in Brazil the involvement of Prototheca alga in bovine mastitis. Treatment with propolis in dimetilsulfoxide (DMSO) resulted in clinical and microbiological cure of 84,8 per cent of treated teats

Aspectos morfológicos e morfométricos da glândula mamária de ratas normais e ooforectomizadas tratadas com metoclopramida / Morphologic and morphometric aspects of the mammary gland of normal and oophorectomized rats treated with metoclopramide

Os autores realizaram, a nível de microscopia óptica comum, estudo morfológico e morfométrico da glândula mamária de ratas na fase de estro e após ooforectomia, tratadas ou näo com metoclopramida. Observaram que no grupo de ratas normais, submetidas à açäo da droga, as glândulas mamárias se apresentavam bem desenvolvidas contendo secreçäo na luz de seus ductos e alvéolos. Já no grupo de ratas ooforectomizadas e tratadas com metoclopramida, as glândulas mamárias se apresentavam atrofiadas, com o mesmo aspecto daquelas da fase de estro. Essas observaçöes estäo de acordo com o estudo morfométrico que revelou um aumento, estatisticamente significante, de parênquima no grupo de ratas normais e tratadas em relaçäo aos demais grupos e, também, maior quantidade de alvéolos com secreçäo