Differentiation of canine adipose mesenchymal stem cells into insulin-producing cells: comparison of different culture medium compositions.

The aim of this study was to differentiate canine adipose-derived mesenchymal stem cells (ADMSCs) into insulin-producing cells by using culture media with different compositions to determine the most efficient media. Stem cells isolated from the fat tissues close to the bitch uterus were distributed into 6 groups: (1) Dulbecco's modified Eagle medium (DMEM)-high glucose (HG), ß-mercaptoethanol, and nicotinamide; (2) DMEM-HG, ß-mercaptoethanol, nicotinamide, and exendin-4; (3) DMEM-HG, ß-mercaptoethanol, nicotinamide, exendin-4, B27, nonessential amino acids, and l-glutamine; (4) DMEM-HG, ß-mercaptoethanol, and nicotinamide (for the initial 8-d period), and DMEM-HG, ß-mercaptoethanol, nicotinamide, exendin-4, B27, nonessential amino acids, l-glutamine, and basic fibroblast growth factor (for the remaining 8-d period); (5) DMEM-HG and fetal bovine serum; and (6) DMEM-low glucose and fetal bovine serum (standard control group). Adipose-derived mesenchymal stem cells from groups 1 to 5 gradually became round in shape and gathered in clusters. These changes differed between the groups. In group 3, the cell clusters were apparently more in numbers and gathered as bigger aggregates. Dithizone staining showed that groups 3 and 4 were similar in terms of the mean area of each aggregate stained for insulin. However, only in group 4, the number of insulin aggregates and the total area of aggregates stained were significantly bigger than in the other groups. The mRNA expression of PDX1, BETA2, MafA, and Insulin were also confirmed in all the groups. We conclude that by manipulating the composition of the culture medium it is possible to induce canine ADMSCs into insulin-producing cells, and the 2-staged protocol that was used promoted the best differentiation.

ω-Conotoxina MVIIC e células-tronco mesenquimais promovem recuperação motora em ratos Wistar após trauma medular agudo / ω-Conotoxin MVIIC and mesenchymal stem cells promote motor recovery in Wistar rats after acute spinal cord injury]

O objetivo deste estudo foi avaliar o efeito da ω-conotoxina MVIIC e das células-tronco mesenquimais (CTM) de forma isolada e sua associação nos ratos submetidos ao trauma medular agudo (TMA). Trinta Rattus novergicus, linhagem Wistar, três meses de idade, foram distribuídos igualmente em cinco grupos experimentais: controle negativo (CN), controle positivo (CP), ω-conotoxina MVIIC (MVIIC), células-tronco mesenquimais da medula óssea (CTM-MO) e associação (MVIIC + CTM-MO). O grupo CN foi submetido à laminectomia sem trauma medular, e os grupos CP, MVIIC, CTM-MO e MVIIC + CTM-MO foram submetidos ao trauma medular contusivo. O grupo CP recebeu, uma hora após o TMA, 10µL de PBS estéril, e os grupos MVIIC e MVIIC + CTM-MO receberam 10µL de PBS contendo 20pmol da ω-conotoxina MVIIC, todos por via intratecal. Os grupos CTM-MO e MVIIC + CTM-MO receberam, 24 horas após, 1x106 de CTM via intravenosa. Avaliou-se a recuperação da função locomotora até o sétimo dia pós-trauma. Os animais tratados com MVIIC + CTM-MO obtiveram recuperação motora após o trauma medular agudo (P<0,05). Conclui-se que essa associação apresentou efeito neuroprotetor com melhora na função locomotora em ratos Wistar.(AU)

The objective of this study was to evaluate the effect of isolated ω-conotoxin MVIIC and mesenchymal stem cells (MSCs) and its association in rats submitted to acute spinal cord injury (SCI). Thirty Rattus norvegicus, Wistar strain, three-month-old rats were randomly distributed in five experimental groups with six animals: negative control (CN), positive control (CP), ω-conotoxin MVIIC (MVIIC), bone marrow mesenchymal stem cells (CTM-MO) and the association (MVIIC + CTM-MO). The CN group underwent laminectomy without spinal cord trauma, and groups CP, MVIIC, CTM-MO and MVIIC + CTM-MO were submitted to contusive spinal cord trauma. The CP group received 10µl of PBS one hour after SCI, and groups MVIIC and MVIIC + CTM-MO received 10µl of PBS containing 20pmol of ω-conotoxin MVIIC, both intrathecally. Groups CTM-MO and MVIIC + CTM-MO received 1x106 of MSCs intravenously 24 hours later. The recovery of locomotor function was evaluated up to seven days post-injury. The animals treated with MVIIC + CTM-MO obtained motor recovery after SCI (P<0.05). It is concluded that this association showed neuroprotective effect with improvements in locomotor function in Wistar rats.(AU)

A Rare Case of Radicular Dens Invaginatus (Dens in Dente) in a Dog.

A rare case of radicular dens invaginatus (dens in dente) was found during dental cleaning of a 5-year-old male Rottweiler dog. Radiographic examination revealed intense radiopacity, which extended from the crown to the apical root region of the affected tooth. Macroscopically, the crown of the left maxillary first molar tooth (209) had irregular and deformed buccal and lingual surfaces. Microscopic examination revealed dentine invagination in the pulp cavity in of the crown and root and pulp necrosis. Based on the gross, radiographic and histological findings, a diagnosis of radicular dens invaginatus was made.

Tumor carcinoide na vesícula biliar em cão / Carcinoid gallbladder tumor in a dog

Carcinoid is a neoplasia that arises from dispersed cells of the neuroendocrine system. This tumor is uncommon in animals, and its occurrence in the gallbladder is rare. A male Basset Hound dog's corpse was taken to the Univerdade Federal de Minas Gerais to be analyzed by the Veterinary Pathology sector, without a description of its previous history. Necropsy revealed the presence of pale oral, ocular and penile mucous membranes. The gallbladder had a thickened wall and a dilated lumen, which was filled with dark and lumpy bile. Its mucosa had a whitish-red nodule, with solid and friable areas. Microscopically, there was a focal neoplastic proliferation, which wasn't encapsulated and had imprecise limits, which cells were distributed in a solid pattern and separated by a delicate fibrovascular stroma. The neoplastic cells presented oval or round shaped nucleus, which had a chromatin predominantly loose, and one or two nucleoli. Their cytoplasm was moderately abundant, and in most of the cells it was eosinophilic, granular, and had well-defined limits. Using the Grimelius coloration, neoplastic cells' cytoplasmic granules stained brownish or black, confirming the neuroendocrine origin of the neoplasia. Based on the macroscopic and microscopic findings, the diagnosis of a gallbladder carcinoid was established.(AU)

Efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e população de células natural killers na decídua e na glândula metrial de ratas / Effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats

Estudou-se o efeito do hipotireoidismo materno na expressão espaço-temporal de mediadores imunológicos e na população de células natural killers (NK) na decídua e na glândula metrial de ratas durante a gestação. Avaliou-se a detecção imunoistoquímica de interferon γ (IFNγ), do fator inibidor de migração (MIF), da interleucina 15 (IL15), do óxido nítrico sintase induzível (iNOS), a marcação com lectina DBA para evidenciação das células NK uterinas DBA+ e a expressão gênica de Ifnγ e Nos2. O hipotireoidismo aumentou o iNOS aos sete dias, a IL15 e o MIF aos 10 e 12 dias, o IFNγ e o MIF aos 14 DG e a expressão dos transcritos gênicos para iNos aos 12 e 19 dias e para Ifnγ aos 14 DG. O hipotireoidismo reduziu a imunomarcação de MIF e lectina DBA aos sete dias, lectina DBA aos 10 e 14 DG, IFNγ aos 12 dias, e a expressão de Ifnγ aos 10 e 19 DG e de iNOS aos 12, 14 e 19 DG, bem como reduziu seus transcritos gênicos aos 10 e 14 DG. Conclui-se que o hipotireoidismo compromete o perfil imunológico na interface materno-fetal ao longo da gestação, particularmente por reduzir o fator anti-inflamatório iNOS e a população de células uNK DBA+.(AU)

The goal of this study was to evaluate the effect of maternal hypothyroidism on the spatiotemporal expression of immunological mediators and population of Natural Killers cells in decidua and metrial gland of rats. Interferon gamma (IFNγ), migration inhibitory factor (MIF), interleukin 15 (IL15), inducible nitric oxide synthase (iNOS), and DBA-Lectin labeling for evidence of uNK DBA+ cells in decidua and genetic expression of Ifnγ and iNos by real-time RT-PCR were evaluated. Hypothyroidism increased protein expression of iNOS at 7 days, IL15 and MIF at 10 and 12 days, IFNγ and MIF at 14 DG in the decidua and/or metrial gland and the gene transcripts for iNOS at 12 and 19 days and for Inf at 14 DG. In addition, hypothyroidism reduced the protein expression of MIF and DBA-Lectin at 7 days, DBA-Lectin at 10 and 14 DG, IFNγ at 12 days, and the gene transcript to Ifnγ at 10 and 19 DGs. Hypothyroidism also reduced the protein expression of iNOS at 12, 14 and 19 DG and reduced its gene transcripts at 10 and 14 DGs. It is concluded that hypothyroidism compromises the immunology profile at the maternal-fetal interface throughout pregnancy, particularly by reducing the anti-inflammatory factor iNOS and population of uNK DBA+ cells.(AU)

Bilateral Primary Obstructive Giant Megaureter in an Adult Dog.

A rare case of bilateral, primary, obstructive, giant megaureter was found during necropsy examination of an 11-year-old female German shepherd dog. On ultrasound examination and at necropsy examination, both ureters were tortuous and extensively dilated with diameter ranging from 1.86 to 4.8 cm. Both vesicoureteral junctions were obstructed by uroliths. A diagnosis of giant megaureter was established using human parameters since these values are not recognized in animals. The classification of obstructive and primary megaureter was determined because the obstruction was due to uroliths at the vesicoureteral junctions.

Direct inhibitory effect of caffeine on viability, synthesis activity and gene expression in cultures of chondrocytes extracted from the articular cartilage of rats / Efeito inibidor direto da cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos da cartilagem articular de ratos

The aim of this study was to evaluate the effect of concentrations of caffeine on the viability, synthesis activity and gene expression in cultures of chondrocytes. Extracted articular cartilage from the femurs and tibias of 15 Wistar rats at three days old to isolate chondrocytes. Chondrocytes were cultured in chondrogenic medium (control) or supplemented with caffeine (0.5, 1.0, 2.0mM). Cell viability, alkaline phosphatase activity and collagen synthesis were assessed using colorimetric assays at 7, 14, 21 days. The chondrocyte cultures of all groups grown under coverslips were stained with hematoxylin-eosin to determine the percentage of cells/field and with PAS, safranin O, alcian blue to determine the percentage of matrix chondrogenic/field at 21 days. The expressions of gene transcripts for aggrecan, collagen-II, Sox-9, Runx-2 and alkaline phosphatase were also evaluated by RT-PCR at 21 days. The means were compared using Student-Newman-Keuls. Caffeine significantly reduced the conversion of MTT to formazan, percentage of cells/field, collagen synthesis, alkaline phosphatase activity, synthesis of PAS+, safranin O+ and alcian blue+ chondrogenic matrix, and the expression of aggrecan, Sox-9 and II collagen. It is concluded that caffeine at concentrations of 0.5, 1.0, 2.0mM has a direct inhibitory effect on chondrogenesis in cultures of chondrocytes from rats.(AU)

O objetivo deste estudo foi avaliar o efeito direto de concentrações de cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos de ratos. As cartilagens dos fêmures e tíbias de 15 ratos Wistar com três dias foram extraídas para isolamento de condrócitos. Os condrócitos foram cultivados em meio condrogênico (controle) ou em meio acrescido de diferentes concentrações de cafeína (0,5, 1,0, 2,0mM). Foram avaliadas a viabilidade celular, a atividade da fosfatase alcalina e a síntese de colágeno por ensaios colorimétricos aos sete, 14 e 21 dias. Condrócitos cultivados sob lamínulas foram corados pela hematoxilina e eosina, para se determinar a porcentagem de células/campo, e pelo PAS, safranina O, alcian Blue, para se determinar a porcentagem de matriz condrogênica/campo aos 21 dias. Foi avaliada a expressão de transcriptos gênicos para Sox-9, Runx-2, agrecano, colágeno-II e fosfatase alcalina por qRT-PCR, aos 21 dias. As médias foram comparadas pelo Student-Newman-Keuls. A cafeína reduziu significativamente o MTT em cristais de formazan, a porcentagem de células/campo, a síntese de colágeno, a atividade da fosfatase alcalina e a síntese de matriz condrogênica PAS+, safranina O+, alcian blue+ e expressão de Sox-9 e colágeno-II. Conclui-se que a cafeína, nas concentrações de 0,5, 1,0, 2,0mM, apresenta efeito inibidor direto sobre a condrogênese em culturas de condrócitos de ratos.(AU)

Um caso raro de destruição total do pâncreas por pancreatite em cão / A rare case of total pancreatic destruction due to pancreatitis in a dog

A 16-year-old female Poodle entered UFMG's Veterinary Hospital with severe prostration, lack of appetite, and vomit. During physical examination, abdominal pain, dehydration, and hyperglycemia (448mg/dl) were found, therefore the animal was admitted under the suspicion of diabetic ketoacidosis. Screening revealed metabolic acidosis, hyperkalemia, glycosuria, ketonuria, and proteinuria. Leukocytosis, thrombocytosis, increase in the number of hepatic enzymes and hyperglycemia were also present in these tests. The ultrasound images showed a smaller and hypoechogenic pancreas, irregularity and folds in duodenum and reactivity of the surrounding tissue, indicating pancreatitis. Thirty days after the dog had been discharged for treatment at home, it was taken back to the veterinary hospital due to status epilepticus, which motivated the owner's decision of euthanasia. During post mortem examination no trace of pancreas was found. On histological examination of the adipose tissue next to the duodenum, only one pancreatic duct was seen, together with inflamatory cells , thus characterizing a rare case of total pancreatic destruction due to pancreatitis.(AU)

Estudo retrospectivo da casuística de felinos domésticos no Hospital Veterinário da Universidade Federal de Minas Gerais, no período de 2005 a 2014 / Retrospective study of the case load of domestic cats at the Veterinary Hospital of the Universidade Federal de Minas Gerais from 2005 to 2014

O objetivo deste estudo foi caracterizar as doenças dos gatos domésticos provenientes dos casos de necropsia e histopatologia do Hospital Veterinário da Universidade Federal de Minas Gerais (HV/UFMG), de 2005 a 2014. Foram analisados 408 exames de necropsia e 197 de biópsias, segundo o sexo, a faixa etária e a raça. Os diagnósticos mais frequentes incluíram doenças infecciosas/inflamatórias ou parasitárias (22,5%), agentes físicos (18,1%), doenças proliferativas (15,2%) e degenerativas (13,5%). Politraumatismo (10,8%) foi responsável pelo maior número de mortes em felinos, sendo as chances três vezes maiores em animais com até 24 meses de idade (P=0,005 OR 3,47 [IC 95%: 1,40-8,57]). Neoplasias epiteliais corresponderam a 26 diagnósticos, sendo 20 (4,9%) casos de malignidade. A ocorrência de carcinoma e seus subtipos foi 18 vezes maior em gatos idosos (P<0,01 OR 18,15 [IC 95%: 7,41-44,45]). A insuficiência renal crônica foi mais frequente em gatos com mais de 120 meses (P=0,01). Machos apresentaram nove vezes mais chances de desenvolver doenças do trato urinário inferior quando comparados às fêmeas (P=0,001 OR 9,50 [IC 95%: 2,78-32,48]). Em relação às biópsias, animais adultos a idosos foram 10 vezes mais representados (P<0,001 OR 10,8 [IC 95%: 3,22-36,79]).(AU)

The aim of this study was to characterize the diseases of domestic cats based on necropsy and histopathological examinations at the Veterinary Hospital of the Universidade Federal de Minas Gerais (HV/UFMG) from 2005 to 2014. A total of 408 necropsy and 197 biopsy samples were analyzed according to gender, age, and breed. The most frequent diagnoses included infectious/inflammatory or parasitic diseases (22.5%), physical agents (18.1%), proliferative (15.2%), and degenerative (13.5%) diseases. Polytrauma (10.8%) was responsible for the highest number of feline deaths, with the odds three times higher in animals up to 24 months of age (P= 0.005 OR 3.47 [95% CI: 1.40-8.57]). Epithelial neoplasms corresponded to 26 diagnoses, with 20 (4.9%) malignant cases. The occurrence of carcinoma and its subtypes was 18-fold higher in older cats (P< 0.01 OR 18.15 [95% CI: 7.41-44.45]). Chronic renal failure was more frequent in cats over 120 months (P= 0.01). Males were nine times more likely to develop lower urinary tract diseases when compared to females (P= 0.001 OR 9.50 [95% CI: 2.78-32.48]). Regarding the biopsies, adult to elderly animals were ten times more represented (P< 0.001 OR 10.8 [95% CI: 3.22-36.79]).(AU)

Osteogenic differentiation of adipose tissue-derived mesenchymal stem cells cultured with different concentrations of prolactin / Diferenciação osteogênica de células tronco mesenquimais do tecido adiposo cultivadas com diferentes concentrações de prolactina

The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.(AU)

O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.(AU)

Excesso de tiroxina materna associado ao hipertireoidismo pós-natal reduz o crescimento ósseo e o perfil proliferativo e angiogênico das cartilagens de crescimento de ratos / Excess maternal thyroxine associated with postnatal hyperthyroidism reduces bone growth and proliferative and angiogenic profile of rats growth cartilage

Foram estudados os efeitos do excesso da tiroxina materna associado ao hipertireoidismo pós-natal sobre o crescimento ósseo e o perfil proliferativo e angiogênico das cartilagens. Dezesseis ratas Wistar adultas foram distribuídas nos grupos tratados com L-tiroxina e controle. A prole do grupo tratado recebeu L-tiroxina do desmame até 40 dias de idade. Ao desmame, foi realizada dosagem plasmática de T4 livre nas mães. Na prole, foram realizados: dosagem plasmática de T3 total e T4 livre, morfometria das tireoides, mensuração do comprimento e largura do fêmur. Nas cartilagens, foi avaliada a expressão imuno-histoquímica e gênica de CDC-47, VEGF, Flk-1, Ang1, Ang2 e Tie2. As médias entre grupos foram comparadas pelo teste T de Student. As concentrações de T4 livre das mães tratadas e de T3 total e T4 livre da prole foram significativamente mais elevadas. A largura do fêmur foi menor nos animais tratados. Houve também redução da imunoexpressão de CDC-47 e de VEGF e dos transcritos gênicos para VEGF e Ang1 nas cartilagens. Conclui-se que o excesso de tiroxina materna associado ao hipertireoidismo pós-natal reduz a largura da diáfise femoral, a proliferação celular e a expressão de VEGF e de Ang1 nas cartilagens de crescimento de ratos.(AU)

The effects of excess of maternal thyroxine associated with postnatal hyperthyroidism at bone growth and proliferative and angiogenic profile of cartilage were studied. Sixteen adult Wistar rats were divided into treated and control groups. The offspring of the treated group received L-thyroxine from weaning to 40 days-old. At weaning, plasma assay of free T4 was measurement on female rats. In the offspring, the following assessments were performed: measurement of total T3 and free T4, histomorphometry analysis of the thyroid, measurement of body weight and length and width of the femur. In femoral growth cartilage, immunostaining of CDC-47, gene or protein expression of VEGF, Flk-1, Ang1, Ang2 and Tie2 were evaluated. Data were analyzed using Student's t-test. Free T4 was significantly higher in treated rats and total T3 and free T4 were significantly higher in offspring. The width of the femur was significantly lower in treated animals. There was lower immunoreactivity of CDC-47, VEGF and lower expression of gene transcripts for VEGF and Ang1. We concluded that the excess maternal thyroxine associated with postnatal hyperthyroidism reduces the width of the femoral shaft, the cell proliferation and gene and protein expression of VEGF and gene expression of Ang1 on the growth cartilage in rats.(AU)

Efeito do produto iônico do biovidro 60S na diferenciação osteogênica de células-tronco mesenquimais do tecido adiposo de cães / Effect of ionic product of bioglass 60S in osteogenic differentiation of mesenchymal stem cells from the adipose tissue of dogs

O objetivo do presente trabalho foi avaliar o produto iônico do biovidro 60S (BV60S) na diferenciação osteogênica de células-tronco mesenquimais de origem adiposa (CTM-AD) de cães. As CTM-AD foram diferenciadas sem OSTe com o produto iônico (PI OST) por sete, 14 e 21 dias. Avaliou-se o MTT, a fosfatase alcalina (FA), o colágeno, mineralização e as expressões de osterix (OSX), sialoproteína óssea (BSP), osteonectina (ON) e osteocalcina (OC). O grupo PI OSTmostrou menor conversão de MTT aos sete dias e maior conversão aos 21 dias. A atividade de FA foi maior no grupo OST, aos 14 e 21 dias. A síntese de colágeno foi maior no grupo OST aos sete e 21 dias. Verificou-se maior área mineralizada no grupo PI OSTem todos os tempos. Não houve diferenças nas expressões de OSX e OC entre os grupos. Observou-se maior expressão de BSP no grupo PI OST, aos 14 e 21 dias. A expressão de ON foi maior no grupo OST aos 14 dias. Concluiu-se que o produto iônico do BV60S favorece a osteogênese in vitro de CTM-AD de cães.

The aim was to evaluate the ionic product of 60S bioglass (BV60S) in osteogenic differentiation of mesenchymal stem cells from adipose tissue (ADMSCs) in dogs. ADMSCs were differentiated without the ionic product (OST) and with the ionic product (PI-OST) for 7, 14 and 21 days. We evaluated the MTT, alkaline phosphatase (ALP), collagen mineralization and expressions of osterix (OSX), bone sialoprotein (BSP), osteonectin (ON) and osteocalcin (OC). The PI-OST group had a lower MTT conversion to 7days and higher conversion at 21 days. The ALP activity was higher in the OST group at 14 and 21 days. Collagen synthesis was higher in the OST group at 7 and 21 days. A higher mineralized area in the PI-OST group was observed at all times. There were no differences in expressions of OSX and OC between groups. We observed increased expression of BSP in the PI-OST group at 14 and 21 days. The expression of ON was higher in the OST group at 14 days. It was concluded that the ionic product of BV60S promotes in vitro osteogenesis of MSC-AD from dogs.

Efeitos da ingestão de cafeína durante a gestação e a lactação sobre a pele de ratas e de filhotes e sua relação com as concentrações séricas do cortisol materno / Effects of caffeine intake during pregnancy and lactation on the skin of rats and offspring and their relationship to maternal serum concentrations of cortisol

O objetivo deste trabalho foi verificar os efeitos da ingestão materna de diferentes doses de cafeína durante a gestação e a lactação, na pele de ratas-mães e filhotes, bem como sua relação com as concentrações séricas do cortisol materno. Vinte e quatro ratas Wistar adultas foram distribuídas em quatro grupos, representados pelo controle e tratados, com cafeína nas doses de 25, 50 e 100mg/kg. Os grupos tratados receberam cafeína por sonda orogástrica durante toda a gestação e a lactação. O controle recebeu água destilada como placebo. Foram avaliados e quantificados os diferentes tipos de folículos pilosos e a espessura da epiderme. A técnica de imuno-histoquímica, com o uso do anticorpo anti-CDC47, foi utilizada para avaliar a proliferação celular da epiderme e dos folículos pilosos das mães. Na mãe, também foram mensurados os níveis séricos de cortisol pela técnica da quimioluminescência. Os dados foram submetidos à análise de variância com comparação das médias pelos testes Kruskal-Wallis e SNK. Nos grupos tratados com cafeína nas doses de 25 e 50mg/kg, tanto as mães quanto seus filhotes apresentaram hipotricose e/ou alopecia focal. Apesar de a frequência de alterações macroscópicas das mães ter sido superior a dos filhotes, nestes as lesões, quando presentes, foram difusas. A análise histológica demonstrou calcinose de folículos pilosos nas mães e nos filhotes. Mas a morfometria somente revelou diferença significativa no número de folículos pilosos das mães, bem como redução significativa da proliferação celular dos folículos pilosos do grupo tratado com 50mg/kg de cafeína. Os níveis de cortisol materno somente foram significativamente elevados no grupo tratado com 100mg/kg de cafeína. Conclui-se que a cafeína ingerida pelas ratas gestantes e lactantes pode causar lesões cutâneas tanto nas mães quanto nos filhotes, caracterizadas por hipotricose e/ou alopecia, independentemente dos níveis séricos do cortisol materno.

The aim of this study was to investigate the effects of maternal caffeine intake during pregnancy and lactation on the skin of rats and their offspring, as well as their relationship to maternal serum levels of cortisol. 24 adult Wistar rats were equally divided into four groups represented by the control and treated with caffeine at doses of 25, 50 and 100mg/kg. The groups received caffeine by orogastric tube during the entire pregnancy and lactation. The control received distilled water as placebo. Different types of hair follicles and the thickness of the epidermis were assessed and quantified. Immunohystochemistry technique using antibody anti-CDC47 was used to evaluate cellular proliferation of the epidermis and hair follicles of the mothers. Also in the mothers, serum levels of cortisol were measured by the chemiluminescence technique. Data were submitted to analysis of variance comparing mediums by Kruskall Wallis Test and SNK. In groups treated with caffeine 25 and 50mg/kg, both mothers and their puppies had focal alopecia and/or hypotrichosis. Despite the higher frequency of macroscopic changes on the mothers, these lesions were diffuse when present on the puppies. Histological analysis showed calcinosis of hair follicles in the mothers and their puppies. But morphometry revealed significant difference in the number of hair follicles from mothers, as well as a significant reduction of cell proliferation of hair follicles in the group treated with 50mg/kg of caffeine. Maternal cortisol levels were significantly elevated in the group treated with 100mg/kg of caffeine. It is concluded that caffeine intake by pregnant and lactating rats can cause skin lesions in both the mothers and their offspring, characterized by alopecia and/or hypotrichosis, regardless of serum levels of maternal cortisol.

In vitro effects of triiodothyronine on gene expression in mouse trophoblast cells.

The objective of the present study was to evaluate the effects of different doses of T3 (10(-4) M, 10(-7) M, 10(-9) M) on the in vitro gene expression of Tpbp, Prl3b1, VEGF, PGF, PL-1, and INFy in mouse trophoblast cells by real-time RT-PCR. Doses of 10(-7) and 10(-9) M T3 increased the mRNA levels of Tpbp, Pl3b1, VEGF, PGF, INFy and PL-1. In contrast, the dose of 10(-4) M reduced the gene expression of PL-1 and VEGF. T3 affected the gene expression of differentiation, hormonal, immune and angiogenic factors in mouse trophoblast cells.

Multiple congenital defects in a newborn foal / Defeitos congênitos múltiplos em um potro recém-nascido

A case of multiple congenital defects in a newborn foal is reported. The animal showed hypoplasia of the left pelvic limb bones, uterus unicornis, congenital diaphragmatic hernia, and unilateral renal and ureteral agenesis. This report includes the macroscopic and microscopic lesions observed in the case...

Relata-se um caso de defeitos congênitos múltiplos em um potro recém-nascido. O animal apresentou hipoplasia dos ossos do membro pélvico esquerdo, útero unicorno, hérnia diafragmática congênita e agenesia renal e ureteral unilateral. Este relato inclui as lesões macroscópicas e microscópicas observadas no caso...

Acute oesophageal necrosis concurrent with Leishmania chagasi infection in a dog.

A case of acute oesophageal necrosis concurrent with Leishmania chagasi infection is reported in a 6-year-old female mixed-breed dog. The report describes clinical signs, gross and microscopical lesions and immunohistochemical findings.

Diffuse heterotopic hair in a canine uterus: case report / Heterotopia de pelo difusa no útero de uma cadela: relato de caso

The present report describes the clinical and pathological findings related to diffuse heterotopic hairs associated with pseudo-placentational endometrial hyperplasia in a canine uterus. Macroscopically, the uterus was intensely enlarged and thicker, and the endometrial surface was irregular and covered with black hairs similar to the hairs of the bitch. Histologically, heterotopic hairs associated with pseudo-placentational endometrial hyperplasia were observed.

O presente relato descreve os achados clínicos e patológicos de uma heterotopia de pelos difusa associada com hiperplasia endometrial pseudo-placentacional em um útero de uma cadela. Macroscopicamente, o útero estava intensamente aumentado de volume e espesso, a superfície do endométrio estava irregular e recoberta com pelos pretos semelhantes aos pelos da cadela. Histologicamente, heterotopia de pelos associada com hiperplasia endometrial pseudoplacentacional foi observada.

Leiomioma ovariano, tumor de células da granulosa e ovário policístico em cadela: uma rara associação / Ovarian leiomyoma, granulosa cell tumor and polycystic ovary in a bitch: a rare association

This case describes a rare case of ovarian leiomyoma associated with granulosa cell tumor and polycystic ovary and in a fifteen-year-old mongrel female dog.

Effects of hypo- and hyperthyroidism on proliferation, angiogenesis, apoptosis and expression of COX-2 in the corpus luteum of female rats.

Although thyroid dysfunction occurs frequently in humans and some animal species, the mechanisms by which hypo- and hyperthyroidism affect the corpus luteum have not been thoroughly elucidated. This study evaluated the levels of proliferative activity, angiogenesis, apoptosis and expression of cyclooxygenase-2 in the corpus luteum of female rats with thyroid dysfunction. These processes may be important in understanding the reproductive changes caused by thyroid dysfunction. A total of 18 adult female rats were divided into three groups (control, hypothyroid and hyperthyroid) with six animals per group. Three months after treatment to induce thyroid dysfunction, the rats were euthanized in the dioestrus phase. The ovaries were collected and immunohistochemically analysed for expression of the cell proliferation marker CDC-47, vascular endothelial growth factor (VEGF), VEGF receptor Flk-1 and cyclooxygenase-2 (COX-2). Apoptosis was evaluated using the TUNEL assay. Hypothyroidism reduced the intensity and area of COX-2 expression in the corpus luteum (p < 0.05), while hyperthyroidism did not alter COX-2 expression in the dioestrus phase. Hypothyroidism significantly reduced the expression of CDC-47 in endothelial cells and pericytes in the corpus luteum, whereas hyperthyroidism did not induce a detectable change in CDC-47 expression (p > 0.05). Hypothyroidism reduced the level of apoptosis in luteal cells (p < 0.05) and increased VEGF expression in the corpus luteum. In contrast, hyperthyroidism increased the level of apoptosis in the corpus luteum (p < 0.05). In conclusion, thyroid dysfunction differentially affects the levels of proliferative activity, angiogenesis and apoptosis and COX-2 expression in the corpus luteum of female rats.

Primary ovarian rhabdomyosarcoma in a dog.

A 10-year-old female English pointer was diagnosed with an ovarian tumour with abdominal metastases. Ultrasonography revealed several nodules of 1-5 cm diameter within the abdominal cavity. Fine needle aspiration cytology of the nodules suggested a malignant mesenchymal tumour. On necropsy examination the right ovary and its capsule were enlarged and there were white-red, friable nodular masses distributed over the surface of the pancreas, liver, omentum, mesentery and serosae of the small and large intestines. Microscopical evaluation revealed neoplastic cells with a high degree of pleomorphism and vascular invasion. Immunohistochemically, the neoplastic cells expressed myosin, desmin, vimentin and CD10, but were negative for cytokeratin, placental alkaline phosphatase, inhibin-alpha and smooth muscle actin. Based on these findings a diagnosis of primary ovarian alveolar rhabdomyosarcoma was made.