GATA-3 expression and its correlation with prognostic factors and survival in canine mammary tumors.

Introduction: The transcription factor GATA-3 plays a significant role in mammary gland development and differentiation. Recent studies on human oncology have demonstrated its association with favorable pathologic factors in breast cancer. Canine mammary tumours, proposed as comparative and translational study models, have epidemiological, clinical, biological, and genetic characteristics similar to those of human breast cancers. Methods: Here, we evaluated the frequency of GATA-3 expression in mammary tumors of dogs and its relationship with prognostic factors and survival. Tumor samples were obtained from 40 female dogs and grouped according to histological type into benign tumors (n = 10), carcinoma in mixed tumors (CMTs) (n = 20), and aggressive tumors (n = 10). CMTs were further separated according to histological grade, and data on clinical staging and diagnosis, histopathological grading, and survival rate were collected. Results: GATA-3 and estrogen receptor (ER) expression were higher in benign and well-differentiated carcinomas than in aggressive tumors, which showed greater Ki-67 expression. The expression rate of ER in the studied groups was equivalent to that of GATA-3. We identified a strong positive correlation between GATA-3 and ER expression frequencies and a negative correlation between those of GATA-3 and Ki-67. There were associations between GATA-3 (p < 0.001), Ki-67 (p = 0.003), tumor size (p < 0.001), clinical stage (p = 0.002), lymph node metastasis (p < 0.001), and histological grade (p < 0.001) by univariate survival analysis. The parameters ER (p = 0.015) and GATA-3 (p = 0.005) also influenced survival in a multifactorial manner. Discussion: Kaplan-Meier analysis of survival curves validated our previous findings that dogs with GATA-3 expression in ≥79.4% of cells had significantly higher survival rates (p < 0.001). The performance analysis showed that the expression of GATA-3 in ≥79.4% of cells effectively predicted survival or death in dogs with mammary tumors. Collectively, these results suggest that GATA-3 can be a relevant marker in the study of mammary tumor progression and has potential as a prognosis marker for predicting outcomes in canine mammary tumors.

Excess Maternal Thyroxine Alters the Proliferative Activity and Angiogenic Profile of Growth Cartilage of Rats at Birth and Weaning.

Objective The aim of this study was to unravel the mechanisms by which thyroxine affects skeletal growth by evaluating proliferative activity and angiogenic profile of growth cartilage of neonatal and weanling rats. Methods Sixteen adult Wistar rats were equally divided into 2 groups: control and treated with thyroxine during pregnancy and lactation. The weight, measurement of plasma free T4 and thyroids, femurs' histomorphometric analysis, and proliferative activity and angiogenic profile by immunohistochemical or real-time reverse transcriptase-polymerase chain reaction in growth cartilage was performed. Data were analyzed using Student's t test. Results The free T4 was significantly higher in the treated rats. However, the height of the follicular epithelium of the thyroid in newborns was significantly lower in the treated group. The excess maternal thyroxine significantly reduced the body weight and length of the femur in the offspring but significantly increased the thickness of trabecular bone and changed the height of the zones of the growth plate. Furthermore, excess maternal thyroxine reduced cell proliferation and vascular endothelial growth factor (VEGF) expression in the growth cartilage of newborn and 20-day-old rats ( P < 0.05). There was also a reduction in the immunohistochemical expression of Tie2 in the cartilaginous epiphysis of the newborns and FLK-1 in the articular cartilage of 20-day-old rats. No significant difference was observed in Ang2 expression. Conclusions The excess maternal thyroxine during pregnancy and lactation reduced endochondral bone growth in the progeny and reduced the proliferation rate and VEGF, Flk-1, and Tie2 expression in the cartilage of growing rats without altering the mRNA expression of Ang1 and Ang2.

Excess maternal and postnatal thyroxine alters chondrocyte numbers and the composition of the extracellular matrix of growth cartilage in rats.

Purpose/Aim: The aim of this study was to evaluate the effects of excess maternal and postnatal thyroxine on chondrocytes and the extracellular matrix (ECM) of growth cartilage. MATERIALS AND METHODS: We used 16 adult female Wistar rats divided into two groups: thyroxine treatment and control. From weaning to 40 days of age, offspring of the treated group (n = 8) received L-thyroxine. Plasma free T4 was measured. Histomorphometric analysis was performed on thyroids and femurs of all offspring. Alcian blue histochemical staining and real-time reverse transcription polymerase chain reaction measurements of gene expression levels of Sox9, Runx2, Aggrecan, Col I, Col II, Alkaline phosphatase, Mmp2, Mmp9, and Bmp2 were performed. Data were analyzed for statistical significance by student's t-test. RESULTS: Excess maternal and postnatal thyroxine reduced the intensity of Alcian blue staining, altered the number of chondrocytes in proliferative and hypertrophic zones in growth cartilage, and reduced the gene expression of Sox9, Mmp2, Mmp9, Col II, and Bmp2 in the growth cartilage of all offspring. Additionally, excess thyroxine altered the gene expression of Runx2, Aggrecan and Col I, and this effect was dependent on age. CONCLUSIONS: Excess thyroxine in neonates suppresses chondrocyte proliferation, stimulates chondrocyte hypertrophy and changes the ECM composition by reducing the amount of proteoglycans and glycosaminoglycans (GAGs). Prolonged exposure to excess thyroxine suppresses chondrocyte activity in general, with a severe reduction in the proteoglycan content of cartilage and the expression of gene transcripts essential for endochondral growth and characteristics of the chondrocyte phenotype.

Effects of excess maternal thyroxin on the bones of rat offspring from birth to the post-weaning period.

Objective To evaluate, in rat offspring, bone changes induced by excess maternal thyroxin during pregnancy and lactation, and to assess the reversibility of these changes after weaning. Material and methods Twenty Wistar rats were distributed in two groups, hyperthyroid and control, that were treated daily with L-thyroxin (50 mcg/animal) and placebo, respectively. The treatment was initiated seven days before mating and continued throughout pregnancy and lactation. From every female of each of the two groups, two offspring were euthanized after birth, two at 21 days of age (weaning), and two at 42 days of age (21 days after weaning). In newborns, the length of pelvic and thoracic limbs were measured, and in the other animals, the length and width of the femur and humerus were measured. Bones were dissected, decalcified, embedded in paraffin, and analyzed histomorphometrically. Results Excess maternal thyroxin significantly reduced the length of the pelvic limb in neonates. In 21-day-old individuals, excess maternal thyroxine reduced the length and the width of the femur and the humerus. It also increased thickness of the epiphyseal plate and the percentage of trabecular bone tissue. In 42-day-old individuals, there were no significant differences between groups in relation to the parameters evaluated in the previous periods. Conclusion Excess maternal thyroxine reduced growth in suckling rats both at birth and at weaning, and it also increased the percentage of trabecular bone tissue in 21-day-old animals. These changes, however, were reversible at 42 days, i.e., 21 days after weaning. Arch Endocrinol Metab. 2016;60(2):130-7.

Effects of excess maternal thyroxin on the bones of rat offspring from birth to the post-weaning period

ABSTRACT Objective To evaluate, in rat offspring, bone changes induced by excess maternal thyroxin during pregnancy and lactation, and to assess the reversibility of these changes after weaning. Material and methods Twenty Wistar rats were distributed in two groups, hyperthyroid and control, that were treated daily with L-thyroxin (50 mcg/animal) and placebo, respectively. The treatment was initiated seven days before mating and continued throughout pregnancy and lactation. From every female of each of the two groups, two offspring were euthanized after birth, two at 21 days of age (weaning), and two at 42 days of age (21 days after weaning). In newborns, the length of pelvic and thoracic limbs were measured, and in the other animals, the length and width of the femur and humerus were measured. Bones were dissected, decalcified, embedded in paraffin, and analyzed histomorphometrically. Results Excess maternal thyroxin significantly reduced the length of the pelvic limb in neonates. In 21-day-old individuals, excess maternal thyroxine reduced the length and the width of the femur and the humerus. It also increased thickness of the epiphyseal plate and the percentage of trabecular bone tissue. In 42-day-old individuals, there were no significant differences between groups in relation to the parameters evaluated in the previous periods. Conclusion Excess maternal thyroxine reduced growth in suckling rats both at birth and at weaning, and it also increased the percentage of trabecular bone tissue in 21-day-old animals. These changes, however, were reversible at 42 days, i.e., 21 days after weaning. Arch Endocrinol Metab. 2016;60(2):130-7.

Osteogenic potential of osteoblasts from neonatal rats born to mothers treated with caffeine throughout pregnancy.

BACKGROUND: Caffeine is an active alkaloid that can cause damage to bones in formation during prenatal life into adulthood. This compound can pass across the placenta and into the mother's milk, causing a reduction in bone formation, growth and mass. The objective of this study was to examine the osteogenic potential of osteoblasts extracted from neonatal rats born to mothers treated with caffeine throughout pregnancy. METHODS: Twenty-four adult Wistar rats were randomly divided into four groups, consisting of one control group and three groups that were treated with 25, 50, or 100 mg/kg of caffeine by an oral-gastric probe throughout the duration of the experimental period (pregnancy). At birth, three puppies from each dam in each group were euthanized, and osteoblasts were extracted from the calvaria of these pups for in vitro testing. RESULTS: The osteoblasts extracted from the pups of rats that received 50 mg/kg caffeine during pregnancy exhibited increased expression of osteocalcin, osteopontin, sialoprotein, runx-2, alkaline phosphatase and type I collagen transcripts, resulting in increased synthesis of mineralization nodules. CONCLUSIONS: Neonates from rats treated with 50 mg/kg caffeine during pregnancy contained osteoblasts with a higher osteogenic potential characterized by increased expression of osteocalcin, osteopontin, sialoprotein, runx-2, alkaline phosphatase and type I collagen and increased synthesis of mineralization nodules.

Understanding of the immunological heterogeneity of canine mammary carcinomas to provide immunophenotypic features of circulating leukocytes as clinically relevant prognostic biomarkers.

We have evaluated the phenotypic features of peripheral blood leukocytes as putative novel biomarkers with prognostic values to monitor canine mammary carcinomas. Female dogs were categorized into distinct groups, referred as mammary carcinoma in benign mixed tumor-MC-BMT and mammary carcinoma-MC. Our findings demonstrate that decreased percentage of B-cells along with increased frequency of NK-cells, CD8(+)T-cells, and CD8(+)CD5(Low+)T-cells beside higher T/B-cells and lower CD4(+)/CD8(+) ratio were the hallmarks of MC-BMT. Despite the lower expression of MHCI and MHCII, the lymphocytes from MC-BMT and MC displayed higher migration potential as suggested by enhanced frequency of CD18(+) events. Although increased levels of macrophage-like cells/(CD14(+)CD16(+)) and decreased levels of MHCII expression were a common phenotypic feature in mammary carcinoma, down-regulation of MHCI was selectively observed in MC. Decreased frequency of CD4(+) T-cells with increased levels of CD8(+) T-cells and lower CD4(+)/CD8(+) T-cell ratio were relevant biomarkers of MC-BTM(-). Although decreased expression of MHCI by monocytes was observed in MC-BTM regardless of the presence of lymph node metastasis, this phenotypic feature was restricted to MC free of metastasis. The CD4(+)/CD8(+) T-cells ratio lower than 1.8 was elected as a valid parameter with outstanding performance to predict survival in MC-BMT. On the other hand, the MHCI expression by monocytes higher than 10(2) MFI showed good value to estimate worse outcome in MC. These results should help to improve our understanding of the immunological heterogeneity of canine mammary carcinomas and provide tools for the determination of cut-off scores of clinically relevant immonophenotypic prognostic biomarkers.

Detecção de aglomerados espaciais de casos de neoplasia mamária em cães no município de Salvador, Bahia / Detection of spatial clusters for breast cancer in canines in the city of Salvador, Bahia

Os tumores mamários espontâneos representam a neoplasia mais frequente em fêmeas caninas, correspondendo aproximadamente a 50 por cento de todas as neoplasias. A maioria dos estudos científicos restringe-se a dados pontuais sobre a doença, sem a preocupação com sua distribuição geográfica ou mesmo com a possibilidade da geração de agregados desses eventos em uma determinada área. Levando-se em consideração a lacuna de informações na literatura, o presente trabalho teve como objetivo a criação de mapas temáticos da distribuição espacial das neoplasias mamárias em cadelas e a identificação de aglomerados de risco para a doença no município de Salvador, Bahia. Pela análise espacial de varredura, verificou-se que os casos de neoplasia mamária não estão homogeneamente distribuídos no município. Foi detectado um aglomerado primário estatisticamente significante (P<0,001), abrangendo 67,3 por cento dos casos estudados. Espera-se que estes resultados contribuam principalmente na elaboração de novos estudos, nos quais devem ser analisadas variáveis de caráter intrínseco e extrínseco ao animal para a identificação dos fatores de risco e elaboração de planos educacionais direcionados à promoção da saúde animal.

Spontaneous mammary tumors represent the most frequent type of cancer in canines, accounting for approximately 50 percent of all neoplasms. The majority of scientific papers cited in the literature are limited to non refined epidemiological data, without mentioning the trend of this disease in generating clusters in a given geographical area. In this context, this research aimed to create thematic maps of spatial distribution of mammary neoplasms in bitches and to identify disease clusters for the city of Salvador, Bahia. Trough the spatial analysis scanning, it was found that cases of breast cancer is not evenly distributed in the municipality. A significant primary cluster was detected (P>0,001) covering 67.3 percent of the studied cases. Considering the gap in literature available in this field, it is believed that such results will become very important, especially in leading to new studies, where intrinsic and extrinsic variables regarding the animal must be taken into consideration and analyzed for factors risk identification to formulate educational plans targeting the promotion of animal welfare.

Ectopia ureteral em cães: relato de dois casos / Ectopia uretral en perros: relato de dos casos / Urethral ectopia in dogs: report of two cases

O presente trabalho descreve dois casos distintos de ectopia ureteral em cães atendidos no Hospital de Medicina Veterinária da Universidade Federal da Bahia. O primeiro relata a presença de ureter ectópico intramural unilateral direito em uma cadela da raça poodle, de aproximadamente um ano de idade, que apresentava como sequela a hidronefrose e hidroureter, sendo necessária a ureteronefrectomia direita para resolução do caso. O segundo caso refere-se à presença de ureter ectópico extramural bilateral em uma cadela da raça Husky Siberiano, de seis anos de idade, cujo prognóstico foi desfavorável em virtude de alterações irreversíveis em todo trato urinário. Em ambos os casos, evidenciou-se a importância do diagnóstico precoce para evitar o desencadeamento de sequelas e garantir a resolução do problema.

This study describes two distinct cases of urethral ectopia in dogs examined at the Veterinary Hospital of the Federal University of Bahia. The first one reports the presence of a right unilateral intramural ectopic ureter in a one-year-old female Poodle that had hydro nephrosis and hydro ureter and that needed right ureter nephrectomy to solve the problem The second case is the presence of a bilateral extramural ectopic ureter in a six-year-old female Siberian Husky whose prognostic was unfavorable due to irreversible alterations throughout the urinary tract. In both cases, it was evident how important an early diagnostic is to avoid sequelae and ensure the problem solution.

Este estudio describe dos casos distintos de ectopia uretral en perros atendidos en el Hospital de Medicina Veterinario de la Universidad Federal de Bahía. El primer caso relata la presencia de uréter ectópico intramural unilateral derecho, en una perra de la raza poodle, de aproximadamente un año de edad, que presentaba como secuela la hidronefrosis e hidrouréter, haciéndose necesario la ureteronefrectomia derecha para resolución del caso. El segundo caso se refiere a presencia de uréter ectópico extramural bilateral en una perra de la raza Husky Siberiano, de seis años de edad, cuyo pronóstico fue desfavorable en virtud de alteraciones irreversibles en todo tracto urinario. En los dos casos, se evidenció la importancia del diagnóstico precoz para evitar el desencadenamiento de secuelas y garantizar la resolución del problema.