Resumo
An eight-year-old castrated female feline was referred for necropsy with a history of apathy, inappetence, abdominal distension, hypersensitivity to abdominal palpation, and evidence of abdominal neoplasms on ultrasound. Macroscopically, multifocal to coalescent, yellowish-white, firm and infiltrative nodules were observed on the surface of the parietal peritonium and in abdominal and thoracic organs. Microscopic characteristics of the neoplasm suggested a neuroendocrine origin, but did not allow a definitive diagnosis and determination of the origin. Immunohistochemistry revealed that neoplastic cells expressed vimentin, CD56, neuron specific enolase, and PGP 9.5 and were negative for biogenic amines and hormonal peptides. Based on anatomopathological and immunohistochemical findings, diagnosis of metastatic non-functional neuroendocrine tumor was confirmed.(AU)
Assuntos
Animais , Gatos , Gatos , Tumores Neuroendócrinos/veterinária , Imuno-Histoquímica , Atrofia de Múltiplos SistemasResumo
An eight-year-old castrated female feline was referred for necropsy with a history of apathy, inappetence, abdominal distension, hypersensitivity to abdominal palpation, and evidence of abdominal neoplasms on ultrasound. Macroscopically, multifocal to coalescent, yellowish-white, firm and infiltrative nodules were observed on the surface of the parietal peritonium and in abdominal and thoracic organs. Microscopic characteristics of the neoplasm suggested a neuroendocrine origin, but did not allow a definitive diagnosis and determination of the origin. Immunohistochemistry revealed that neoplastic cells expressed vimentin, CD56, neuron specific enolase, and PGP 9.5 and were negative for biogenic amines and hormonal peptides. Based on anatomopathological and immunohistochemical findings, diagnosis of metastatic non-functional neuroendocrine tumor was confirmed.
Assuntos
Animais , Gatos , Gatos , Imuno-Histoquímica , Tumores Neuroendócrinos/veterinária , Atrofia de Múltiplos SistemasResumo
In forensic toxicology, the detection of toxic chemicals from human bone marrow is often used in cases with an extended post mortem interval; however, in veterinary medicine, this practice is not used. Therefore, this study was performed to investigate the suitability of bone marrow for toxicological analysis in dogs and cats. Six animals with suspected poisoning were selected; the carcasses were sent for necropsy, and the organs were collected and preserved in buffered formalin and processed routinely for histological examination. In addition, bone marrow samples from the femur, humerus, and tibia were collected for toxicological analysis by liquid chromatography coupled to mass spectrometry detection (LC-MS). This analysis confirmed the presence of aldicarb, aldicarb sulfone, asulam, carbendazim, chlorpyrifos, dichlorvos, thifensulfuron methyl and trifloxysulfuron-sodium and associated with clinical symptoms and anatomo-histopathological alterations it was recognized the poisonings. It is expected that this study will promote the toxicological investigation of bone marrow and open avenues for the use of this tissue as an option for the detection of toxic chemicals in cases of forensic pathology.(AU)
Na toxicologia forense, a detecção de substâncias químicas tóxicas provenientes da medula óssea humana é frequentemente usada em casos com intervalo post mortem prolongado; no entanto, na medicina veterinária, essa prática não é utilizada. Portanto, este estudo foi realizado para investigar a utilização da medula óssea nas análises toxicológicas em cães e gatos. Seis animais com suspeita de intoxicação foram selecionados; as carcaças foram enviadas para necropsia e os órgãos foram coletados e preservados em formalina tamponada e processados rotineiramente para exame histológico. Amostras de medula óssea de fêmur, úmero e tíbia foram coletadas para análise toxicológica por cromatografia líquida acoplada à espectrometria de massa-massa (LC-MS). A análise por LC-MS confirmou a presença dos agrotóxicos aldicarbe, aldicarbe sulfona, asulam, carbendazim, clorpirifós, diclorvós, tifensulfuron metil e trifloxisulfuron-sódico, e em associação com sinais clínicos e achados anatomo-histopatológicos comprovou-se as intoxicações. Espera-se que este estudo promova a utilização da medula óssea como uma opção na investigação toxicológica para a detecção de produtos químicos tóxicos em casos de patologia forense.(AU)
Assuntos
Animais , Gatos , Cães , Patologia Veterinária , Intoxicação/diagnóstico , Medula Óssea , Agroquímicos/intoxicação , Substâncias Tóxicas , Intoxicação por Organofosfatos/diagnóstico , Herbicidas/intoxicação , Diclorvós , Clorpirifos , Intoxicação por Organofosfatos/veterináriaResumo
In forensic toxicology, the detection of toxic chemicals from human bone marrow is often used in cases with an extended post mortem interval; however, in veterinary medicine, this practice is not used. Therefore, this study was performed to investigate the suitability of bone marrow for toxicological analysis in dogs and cats. Six animals with suspected poisoning were selected; the carcasses were sent for necropsy, and the organs were collected and preserved in buffered formalin and processed routinely for histological examination. In addition, bone marrow samples from the femur, humerus, and tibia were collected for toxicological analysis by liquid chromatography coupled to mass spectrometry detection (LC-MS). This analysis confirmed the presence of aldicarb, aldicarb sulfone, asulam, carbendazim, chlorpyrifos, dichlorvos, thifensulfuron methyl and trifloxysulfuron-sodium and associated with clinical symptoms and anatomo-histopathological alterations it was recognized the poisonings. It is expected that this study will promote the toxicological investigation of bone marrow and open avenues for the use of this tissue as an option for the detection of toxic chemicals in cases of forensic pathology.(AU)
Na toxicologia forense, a detecção de substâncias químicas tóxicas provenientes da medula óssea humana é frequentemente usada em casos com intervalo post mortem prolongado; no entanto, na medicina veterinária, essa prática não é utilizada. Portanto, este estudo foi realizado para investigar a utilização da medula óssea nas análises toxicológicas em cães e gatos. Seis animais com suspeita de intoxicação foram selecionados; as carcaças foram enviadas para necropsia e os órgãos foram coletados e preservados em formalina tamponada e processados rotineiramente para exame histológico. Amostras de medula óssea de fêmur, úmero e tíbia foram coletadas para análise toxicológica por cromatografia líquida acoplada à espectrometria de massa-massa (LC-MS). A análise por LC-MS confirmou a presença dos agrotóxicos aldicarbe, aldicarbe sulfona, asulam, carbendazim, clorpirifós, diclorvós, tifensulfuron metil e trifloxisulfuron-sódico, e em associação com sinais clínicos e achados anatomo-histopatológicos comprovou-se as intoxicações. Espera-se que este estudo promova a utilização da medula óssea como uma opção na investigação toxicológica para a detecção de produtos químicos tóxicos em casos de patologia forense.(AU)
Assuntos
Animais , Gatos , Cães , Patologia Veterinária , Intoxicação/diagnóstico , Medula Óssea , Agroquímicos/intoxicação , Substâncias Tóxicas , Intoxicação por Organofosfatos/diagnóstico , Herbicidas/intoxicação , Diclorvós , Clorpirifos , Intoxicação por Organofosfatos/veterináriaResumo
A two-year-old, male, non-castrated cat was referred to the veterinary hospital with a three-month history of paresis of the pelvic limbs. Clinical examination revealed a discreet muffling of cardiac sounds and the presence of a degree III/VI holosystolic murmur. The animal was hospitalized and died of cardiorespiratory arrest. The heart had a thickened left ventricular endocardium characterized by the deposition of a fibrous white matter that was firmly adhered to the endocardium and also observed on the epicardium. Renal infarctions and thrombi in the atrium and bifurcation of the aorta were found. Microscopically, in the endocardium of the left ventricular chamber there was a diffuse and disorganized deposition of fibrous connective tissue filled with elastic fibers of varying thickness. In the left ventricular epicardium, the same lesion was observed, but with less extension. Clinical and anatomopathological findings were consistent with primary left ventricular endocardial and left ventricular epicardial fibroelastosis with secondary left ventricular congestive heart failure, thromboembolism and paresis.
Assuntos
Animais , Gatos , Fibroelastose Endocárdica/complicações , Fibroelastose Endocárdica/patologia , Fibroelastose Endocárdica/veterinária , Paresia/veterinária , Tromboembolia/veterinária , Insuficiência Cardíaca/veterinária , Parada Cardíaca/veterináriaResumo
A two-year-old, male, non-castrated cat was referred to the veterinary hospital with a three-month history of paresis of the pelvic limbs. Clinical examination revealed a discreet muffling of cardiac sounds and the presence of a degree III/VI holosystolic murmur. The animal was hospitalized and died of cardiorespiratory arrest. The heart had a thickened left ventricular endocardium characterized by the deposition of a fibrous white matter that was firmly adhered to the endocardium and also observed on the epicardium. Renal infarctions and thrombi in the atrium and bifurcation of the aorta were found. Microscopically, in the endocardium of the left ventricular chamber there was a diffuse and disorganized deposition of fibrous connective tissue filled with elastic fibers of varying thickness. In the left ventricular epicardium, the same lesion was observed, but with less extension. Clinical and anatomopathological findings were consistent with primary left ventricular endocardial and left ventricular epicardial fibroelastosis with secondary left ventricular congestive heart failure, thromboembolism and paresis.(AU)
Assuntos
Animais , Gatos , Paresia/veterinária , Fibroelastose Endocárdica/veterinária , Tromboembolia/veterinária , Fibroelastose Endocárdica/complicações , Fibroelastose Endocárdica/patologia , Parada Cardíaca/veterinária , Insuficiência Cardíaca/veterináriaResumo
A 9-year-old, 5.5-kg (12.12-lb) spayed female Poodle was evaluated due to recurrent seizures and possible diabetes. The dog also had a history of anorexia, polyuria and polydipsia. Serum biochemistry results revealed increases in pancreatic amylase and glucose associated with decreased protein levels. Abdominal ultrasound suggested acute pancreatitis and/or a pancreatic tumor with intense peripancreatic vascularization. The clinical condition of the dog worsened despite medical treatment, and death occurred shortly thereafter. At necropsy, blood clots were found in the stomach and in the intestinal lumen, which was filled with undigested blood. The pancreas was moderately increased in size with an irregular surface and multifocal yellow firm areas. In the cranial portion of the pancreas, a single nodule, approximately 2 cm in diameter, was found. Histologic evaluation of the pancreas revealed chronic and acute lesions characterized by lymphohistiocytic pancreatitis, periductal fibrosis, degeneration of pancreatic islets and necrotizing and fibrinopurulent pancreatitis. The nodule was diagnosed as pancreatic adenocarcinoma. These clinical, imaging, and histopathologic findings were compatible with Hemosuccus pancreaticus. To the authors knowledge, this is the first report of Hemosuccus pancreaticusin a dog and it should be considered as a differential diagnosis for dogs with pancreatic disease and gastrointestinal bleeding.
Assuntos
Animais , Cães , Glucose/análise , Hemorragia Gastrointestinal/veterinária , Pancreatite/veterinária , alfa-Amilases Pancreáticas/análise , Evolução FatalResumo
A 9-year-old, 5.5-kg (12.12-lb) spayed female Poodle was evaluated due to recurrent seizures and possible diabetes. The dog also had a history of anorexia, polyuria and polydipsia. Serum biochemistry results revealed increases in pancreatic amylase and glucose associated with decreased protein levels. Abdominal ultrasound suggested acute pancreatitis and/or a pancreatic tumor with intense peripancreatic vascularization. The clinical condition of the dog worsened despite medical treatment, and death occurred shortly thereafter. At necropsy, blood clots were found in the stomach and in the intestinal lumen, which was filled with undigested blood. The pancreas was moderately increased in size with an irregular surface and multifocal yellow firm areas. In the cranial portion of the pancreas, a single nodule, approximately 2 cm in diameter, was found. Histologic evaluation of the pancreas revealed chronic and acute lesions characterized by lymphohistiocytic pancreatitis, periductal fibrosis, degeneration of pancreatic islets and necrotizing and fibrinopurulent pancreatitis. The nodule was diagnosed as pancreatic adenocarcinoma. These clinical, imaging, and histopathologic findings were compatible with Hemosuccus pancreaticus. To the authors knowledge, this is the first report of Hemosuccus pancreaticusin a dog and it should be considered as a differential diagnosis for dogs with pancreatic disease and gastrointestinal bleeding.(AU)
Assuntos
Animais , Cães , Hemorragia Gastrointestinal/veterinária , Pancreatite/veterinária , alfa-Amilases Pancreáticas/análise , Glucose/análise , Evolução FatalResumo
This study aimed to evaluate the effects of thyroid hormone on the decidua and metrial gland of rats and to examine the expression of angiogenic factors. 72 adult, female rats were divided into hypothyroid, T4-treated2, and control groups. At 10, 14 and 19 days of gestation (DG), the decidua and metrial gland were collected for histomorphometric and immunohistochemical evaluation of the expression of VEGF, Flk-1 and Tie-2. Hypothyroidism reduced the area of the decidua at 10 and 19 DG. Furthermore, VEGF was increased at 10 and 14 DG, and Flk-1 only at 14 DG, but both was reduced at 19 DG in the metrial gland without significantly changing the area occupied by blood vessels. Rats treated with T4 showed an increase in the decidua blood vessels at 10 and 19 DG. However, at 10 DG, excess T4 resulted in increased of Flk-1 in the decidua and metrial gland. Hypothyroidism increased the Tie-2 at 10 and 19 DG in the decidua and metrial gland. In conclusion, hypothyroidism reduces the area of the decidua and increases the expression of VEGF, Tie-2 and Flk-1. The excess of T4 promotes tissue angiogenesis by increasing the number of vessels in the decidua because of the increased expression of Flk-1.(AU)
Este estudo teve como objetivo avaliar os efeitos dos hormônios tireoidianos sobre a decídua e a glândula metrial pela análise da expressão de fatores angiogênicos em ratas. 72 ratas adultas, fêmeas foram distribuídas nos grupos hipotiroideo, tratado com T4 e controle. Aos 10, 14 e 19 dias de gestação (DG), a decídua e a glândula metrial foram coletadas para avaliação histomorfométrica e imunoistoquímica da expressão de VEGF, Flk-1 e Tie-2. O hipotireoidismo reduziu a área da decídua aos 10 e 19 DG. Além disso, o VEGF aumentou aos 10 e 14 DG e o Flk-1 apenas aos 14 DG, mas ambos foram reduzidos aos 19 DG na glândula metrial sem alterar significativamente a área ocupada pelos vasos sanguíneos. As ratas tratadas com T4 apresentaram aumento do número de vasos sanguíneos na decídua aos 10 e 19 DG. Além disso, aos 10 DG, o excesso de T4 resultou no aumento de Flk-1 na decídua e na glândula metrial. O hipotireoidismo aumentou o Tie-2 em 10 e 19 DG na decídua e na glândula metrial. Desta forma, pode-se concluir que o hipotireoidismo reduz a área da decídua e aumenta a expressão de VEGF, Tie-2 e Flk-1. O excesso de T4 promove a angiogênese tecidual ao aumentar o número de vasos na decídua devido ao aumento da expressão de Flk-1.(AU)
Assuntos
Animais , Feminino , Ratos , Feniltioureia/análise , Hormônios Tireóideos/análise , Decídua , Indutores da Angiogênese/análise , Glândula MetrialResumo
This study aimed to evaluate the effects of thyroid hormone on the decidua and metrial gland of rats and to examine the expression of angiogenic factors. 72 adult, female rats were divided into hypothyroid, T4-treated2, and control groups. At 10, 14 and 19 days of gestation (DG), the decidua and metrial gland were collected for histomorphometric and immunohistochemical evaluation of the expression of VEGF, Flk-1 and Tie-2. Hypothyroidism reduced the area of the decidua at 10 and 19 DG. Furthermore, VEGF was increased at 10 and 14 DG, and Flk-1 only at 14 DG, but both was reduced at 19 DG in the metrial gland without significantly changing the area occupied by blood vessels. Rats treated with T4 showed an increase in the decidua blood vessels at 10 and 19 DG. However, at 10 DG, excess T4 resulted in increased of Flk-1 in the decidua and metrial gland. Hypothyroidism increased the Tie-2 at 10 and 19 DG in the decidua and metrial gland. In conclusion, hypothyroidism reduces the area of the decidua and increases the expression of VEGF, Tie-2 and Flk-1. The excess of T4 promotes tissue angiogenesis by increasing the number of vessels in the decidua because of the increased expression of Flk-1.(AU)
Este estudo teve como objetivo avaliar os efeitos dos hormônios tireoidianos sobre a decídua e a glândula metrial pela análise da expressão de fatores angiogênicos em ratas. 72 ratas adultas, fêmeas foram distribuídas nos grupos hipotiroideo, tratado com T4 e controle. Aos 10, 14 e 19 dias de gestação (DG), a decídua e a glândula metrial foram coletadas para avaliação histomorfométrica e imunoistoquímica da expressão de VEGF, Flk-1 e Tie-2. O hipotireoidismo reduziu a área da decídua aos 10 e 19 DG. Além disso, o VEGF aumentou aos 10 e 14 DG e o Flk-1 apenas aos 14 DG, mas ambos foram reduzidos aos 19 DG na glândula metrial sem alterar significativamente a área ocupada pelos vasos sanguíneos. As ratas tratadas com T4 apresentaram aumento do número de vasos sanguíneos na decídua aos 10 e 19 DG. Além disso, aos 10 DG, o excesso de T4 resultou no aumento de Flk-1 na decídua e na glândula metrial. O hipotireoidismo aumentou o Tie-2 em 10 e 19 DG na decídua e na glândula metrial. Desta forma, pode-se concluir que o hipotireoidismo reduz a área da decídua e aumenta a expressão de VEGF, Tie-2 e Flk-1. O excesso de T4 promove a angiogênese tecidual ao aumentar o número de vasos na decídua devido ao aumento da expressão de Flk-1.(AU)
Assuntos
Animais , Feminino , Ratos , Feniltioureia/análise , Hormônios Tireóideos/análise , Decídua , Indutores da Angiogênese/análise , Glândula MetrialResumo
ABSTRACT: This study aimed to evaluate the effects of thyroid hormone on the decidua and metrial gland of rats and to examine the expression of angiogenic factors. 72 adult, female rats were divided into hypothyroid, T4-treated2, and control groups. At 10, 14 and 19 days of gestation (DG), the decidua and metrial gland were collected for histomorphometric and immunohistochemical evaluation of the expression of VEGF, Flk-1 and Tie-2. Hypothyroidism reduced the area of the decidua at 10 and 19 DG. Furthermore, VEGF was increased at 10 and 14 DG, and Flk-1 only at 14 DG, but both was reduced at 19 DG in the metrial gland without significantly changing the area occupied by blood vessels. Rats treated with T4 showed an increase in the decidua blood vessels at 10 and 19 DG. However, at 10 DG, excess T4 resulted in increased of Flk-1 in the decidua and metrial gland. Hypothyroidism increased the Tie-2 at 10 and 19 DG in the decidua and metrial gland. In conclusion, hypothyroidism reduces the area of the decidua and increases the expression of VEGF, Tie-2 and Flk-1. The excess of T4 promotes tissue angiogenesis by increasing the number of vessels in the decidua because of the increased expression of Flk-1.
RESUMO: Este estudo teve como objetivo avaliar os efeitos dos hormônios tireoidianos sobre a decídua e a glândula metrial pela análise da expressão de fatores angiogênicos em ratas. 72 ratas adultas, fêmeas foram distribuídas nos grupos hipotiroideo, tratado com T4 e controle. Aos 10, 14 e 19 dias de gestação (DG), a decídua e a glândula metrial foram coletadas para avaliação histomorfométrica e imunoistoquímica da expressão de VEGF, Flk-1 e Tie-2. O hipotireoidismo reduziu a área da decídua aos 10 e 19 DG. Além disso, o VEGF aumentou aos 10 e 14 DG e o Flk-1 apenas aos 14 DG, mas ambos foram reduzidos aos 19 DG na glândula metrial sem alterar significativamente a área ocupada pelos vasos sanguíneos. As ratas tratadas com T4 apresentaram aumento do número de vasos sanguíneos na decídua aos 10 e 19 DG. Além disso, aos 10 DG, o excesso de T4 resultou no aumento de Flk-1 na decídua e na glândula metrial. O hipotireoidismo aumentou o Tie-2 em 10 e 19 DG na decídua e na glândula metrial. Desta forma, pode-se concluir que o hipotireoidismo reduz a área da decídua e aumenta a expressão de VEGF, Tie-2 e Flk-1. O excesso de T4 promove a angiogênese tecidual ao aumentar o número de vasos na decídua devido ao aumento da expressão de Flk-1.
Resumo
O objetivo deste estudo foi comparar o potencial osteogênico das células tronco mesenquimais extraídas da medula óssea (CTM-MO) com as do tecido adiposo (CTM-AD) de cães adultos. As células foram caracterizadas fenotipicamente quanto à expressão de CD29, CD90, CD34 e CD45 e submetidas à diferenciação adipogênica e condrogênica por 21 dias e osteogênica por 7, 14 e 21 dias. Foram constituídos quatro grupos: 1) CTM-MO em meio osteogênico, 2) CTM-MO em meio basal, 3) CTM-AD em meio osteogênico e 4) CTM-AD em meio basal. Aos 7, 14 e 21 dias de diferenciação osteogênica as culturas foram submetidas às avaliações da conversão de MTT em formazan, da atividade da fosfatase alcalina (FA), da síntese de colágeno e de matriz mineralizada, avaliação do número de células por campo e foram quantificados os transcritos gênicos para osterix, sialoproteina óssea (BSP), osteonectina (ON) e osteocalcina (OC). Tanto as células extraídas da medula óssea quanto do tecido adiposo mostraram elevada expressão de marcadores para células tronco e baixa expressão de marcadores de células hematopoiéticas (menor que 2%). Além disso, foram capazes de se diferenciar em osteoblastos, condrócitos e adipócitos. As CTM-AD submetidas à diferenciação osteogênica mostraram maior conversão do MTT em formazan que as CTM-MO, sob mesmas condições aos 7 e 21 dias. O número de células por campo, a atividade da FA, a síntese de colágeno e de matriz mineralizada foram superior nas CTM-AD em diferenciação, em relação às CTM-MO sob as mesmas condições, em todos os tempos estudados. As expressões de osterix, BSP e OC foram predominantemente superiores nas CTM-MO diferenciadas, mas a expressão de ON foi superior nas CTM-AD diferenciadas aos 7, 14 e 21 dias. Conclui-se que as CTM-AD apresentam maior potencial osteogênico que as CTM-MO quando extraídas de cães adultos.(AU)
The aim of this study was to compare the osteogenic potential of mesenchymal stem cells obtained from bone marrow (BM-MSC) with those extracted from adipose tissue (AT-MSC) of adult dogs. The cells were phenotypically categorized according to the expression of CD29, CD90, CD34 and CD45, and submitted to adipogenic and chondrogenic differentiation for 21 days and osteogenic differentiation for 7, 14 and 21 days. Four groups were formed: BM-MSC in osteogenic medium (1), BM-MSC in basal medium (2), AT-MSC in osteogenic medium (3) and ATMSC in basal medium (4). On days 7, 14 and 21 of osteogenic differentiation, the cultures were submitted to evaluations of MTT conversion in formazan, of alkaline phosphatase activity (AP), of collagen and mineralized matrix synthesis, evaluation of the number of cells per field and there was quantification of the gene transcripts for osterix, bone sialoprotein (BSP), osteonectin (ON) and osteocalcin (OC). Both the cells obtained from bone marrow and those from adipose tissue showed high expression of stem cells markers and low expression of hematopoietic cells markers (lower than 2%). Besides, they were able to differentiate into osteoblasts, chondrocytes and adipocytes. AT-MSC submitted to osteogenic differentiation showed higher MTT conversion in formazan than BM-MSC, under the same conditions on days 7 and 21. The number of cells per field, the AP activity, the collagen and mineralized matrix synthesis were higher in AT-MSC en differentiation, in relation to BM-MSC under the same conditions in all evaluated times. Expressions of osterix, BSP and OC were predominantly higher in differentiated BMMSC, however the expression of ON was higher AT-MSC differentiated on days 7, 14 and 21. In conclusion, AT-MSC present higher osteogenic potential than BM-MSC when extracted from adult dogs.(AU)
Assuntos
Animais , Cães , Tecido Adiposo/citologia , Células da Medula Óssea , Osteogênese , Células-Tronco , Regeneração ÓsseaResumo
O objetivo deste estudo foi comparar o potencial osteogênico das células tronco mesenquimais extraídas da medula óssea (CTM-MO) com as do tecido adiposo (CTM-AD) de cães adultos. As células foram caracterizadas fenotipicamente quanto à expressão de CD29, CD90, CD34 e CD45 e submetidas à diferenciação adipogênica e condrogênica por 21 dias e osteogênica por 7, 14 e 21 dias. Foram constituídos quatro grupos: 1) CTM-MO em meio osteogênico, 2) CTM-MO em meio basal, 3) CTM-AD em meio osteogênico e 4) CTM-AD em meio basal. Aos 7, 14 e 21 dias de diferenciação osteogênica as culturas foram submetidas às avaliações da conversão de MTT em formazan, da atividade da fosfatase alcalina (FA), da síntese de colágeno e de matriz mineralizada, avaliação do número de células por campo e foram quantificados os transcritos gênicos para osterix, sialoproteina óssea (BSP), osteonectina (ON) e osteocalcina (OC). Tanto as células extraídas da medula óssea quanto do tecido adiposo mostraram elevada expressão de marcadores para células tronco e baixa expressão de marcadores de células hematopoiéticas (menor que 2%). Além disso, foram capazes de se diferenciar em osteoblastos, condrócitos e adipócitos. As CTM-AD submetidas à diferenciação osteogênica mostraram maior conversão do MTT em formazan que as CTM-MO, sob mesmas condições aos 7 e 21 dias. O número de células por campo, a atividade da FA, a síntese de colágeno e de matriz mineralizada foram superior nas CTM-AD em diferenciação, em relação às CTM-MO sob as mesmas condições, em todos os tempos estudados. As expressões de osterix, BSP e OC foram predominantemente superiores nas CTM-MO diferenciadas, mas a expressão de ON foi superior nas CTM-AD diferenciadas aos 7, 14 e 21 dias. Conclui-se que as CTM-AD apresentam maior potencial osteogênico que as CTM-MO quando extraídas de cães adultos.(AU)
The aim of this study was to compare the osteogenic potential of mesenchymal stem cells obtained from bone marrow (BM-MSC) with those extracted from adipose tissue (AT-MSC) of adult dogs. The cells were phenotypically categorized according to the expression of CD29, CD90, CD34 and CD45, and submitted to adipogenic and chondrogenic differentiation for 21 days and osteogenic differentiation for 7, 14 and 21 days. Four groups were formed: BM-MSC in osteogenic medium (1), BM-MSC in basal medium (2), AT-MSC in osteogenic medium (3) and ATMSC in basal medium (4). On days 7, 14 and 21 of osteogenic differentiation, the cultures were submitted to evaluations of MTT conversion in formazan, of alkaline phosphatase activity (AP), of collagen and mineralized matrix synthesis, evaluation of the number of cells per field and there was quantification of the gene transcripts for osterix, bone sialoprotein (BSP), osteonectin (ON) and osteocalcin (OC). Both the cells obtained from bone marrow and those from adipose tissue showed high expression of stem cells markers and low expression of hematopoietic cells markers (lower than 2%). Besides, they were able to differentiate into osteoblasts, chondrocytes and adipocytes. AT-MSC submitted to osteogenic differentiation showed higher MTT conversion in formazan than BM-MSC, under the same conditions on days 7 and 21. The number of cells per field, the AP activity, the collagen and mineralized matrix synthesis were higher in AT-MSC en differentiation, in relation to BM-MSC under the same conditions in all evaluated times. Expressions of osterix, BSP and OC were predominantly higher in differentiated BMMSC, however the expression of ON was higher AT-MSC differentiated on days 7, 14 and 21. In conclusion, AT-MSC present higher osteogenic potential than BM-MSC when extracted from adult dogs.(AU)
Assuntos
Animais , Cães , Osteogênese , Células-Tronco Adultas , Tecido Adiposo/citologia , Células da Medula Óssea , Achados Morfológicos e Microscópicos , Regeneração ÓsseaResumo
Objective: Evaluate the effect of in vitro triiodothyronine (T3) on the reduced osteogenic potential of bone marrow mesenchymal stem cells (BMMSCs) of adult rats with osteoporosis compared with BMMSCs of young and adult rats without osteoporosis. Methods: groups were tested: BMMSCs of young rats; BMMSCs of adult rats without osteoporosis; BMMSCs of adult rats with osteoporosis without T3 and BMMSCs of adult rats with osteoporosis treated with T3 (0.01, 1, 100 and 1000 nM). Alkaline phosphatase activity, MTT reduction, mineralized nodules and gene expression for collagen, osteocalcin, sialoprotein, osteopontin and BMP-2 were evaluated. Results: Osteoporosis increased the alkaline phosphatase activity and reduced the formation of mineralized nodules and expression of collagen and osteopontin in at least one of the observed time points. However, the T3 treatment of BMMSCs of rats with osteoporosis altered these parameters. Conclusion: It was concluded that doses of T3, 0.01 and 1000 nM had a positive effect promoted by increased osteogenic matrix synthesis and collagen expression in at least one of the evaluated time points compared to BMMSCs of rats with osteoporosis without treatment. However, T3 was unable to reach the osteogenic potential of the MSCs of healthy young rats.(AU)
Assuntos
Cobaias , Ratos , Osteoporose/veterinária , Medula Óssea , Hormônios Tireóideos/uso terapêutico , Osteogênese , Células-Tronco , Técnicas de Cultura de Células/veterináriaResumo
Objective: Evaluate the effect of in vitro triiodothyronine (T3) on the reduced osteogenic potential of bone marrow mesenchymal stem cells (BMMSCs) of adult rats with osteoporosis compared with BMMSCs of young and adult rats without osteoporosis. Methods: groups were tested: BMMSCs of young rats; BMMSCs of adult rats without osteoporosis; BMMSCs of adult rats with osteoporosis without T3 and BMMSCs of adult rats with osteoporosis treated with T3 (0.01, 1, 100 and 1000 nM). Alkaline phosphatase activity, MTT reduction, mineralized nodules and gene expression for collagen, osteocalcin, sialoprotein, osteopontin and BMP-2 were evaluated. Results: Osteoporosis increased the alkaline phosphatase activity and reduced the formation of mineralized nodules and expression of collagen and osteopontin in at least one of the observed time points. However, the T3 treatment of BMMSCs of rats with osteoporosis altered these parameters. Conclusion: It was concluded that doses of T3, 0.01 and 1000 nM had a positive effect promoted by increased osteogenic matrix synthesis and collagen expression in at least one of the evaluated time points compared to BMMSCs of rats with osteoporosis without treatment. However, T3 was unable to reach the osteogenic potential of the MSCs of healthy young rats.
Assuntos
Cobaias , Ratos , Hormônios Tireóideos/uso terapêutico , Medula Óssea , Osteogênese , Osteoporose/veterinária , Células-Tronco , Técnicas de Cultura de Células/veterináriaResumo
Morphological and immunohistochemical characterization of angiogenic and apoptotic factors and the expression of thyroid receptors in the ovary of tilapia Oreochromis niloticus in captivity were studied. The morphological evaluation of the ovaries was performed by histological paraffin embedded and stained with HE. The immunohistochemical expressions of CDC47, VEGF, Flk-1, angiopoietin, Tie-2 and thyroid receptor (TRα) were performed by the technique of streptavidein-biotin-peroxidase. Apoptosis was assessed using the TUNEL kit. The relative expression of thyroid hormone receptors (TRα and TRß) was assessed by RT-PCR real time. The nuclear expression of CDC47 increased with the stage of maturation of the oocyte and was observed in the follicle cells. Apoptotic bodies were observed in the follicular cells of atretic follicles and postovulatory follicles from the ovaries of 150g and 350g fish. Expression of VEGF and its receptor Flk-1 was also observed in the follicular cells, and the expression of both increased with the maturity of the oocyte, with a higher intensity observed in the full-grown follicle. The expression of angiopoietin and of its receptor (Tie 2) was discrete and moderate respectively. TRα expression was independent of follicular development. However, the 350 g tilapia exhibited higher expression of TRß compared with the 50 g tilapia. We conclude that the proliferative activity and the expression of VEGF and its receptor increase with follicular maturation and that the TRs expression increases with ovarian maturity in tilapia (Oreochromis niloticus).(AU)
Foram estudadas as caracterizações morfológica e imuno-histoquímica de fatores angiogênicos e apoptóticos e a expressão de receptores tireoidianos no ovário de tilápia Oreochromis niloticus de cativeiro. A avaliação morfológica dos ovários foi realizada por cortes histológicos incluídos em parafina e corados por HE. As expressões imuno-histoquímicas de CDC47, VEGF e seu receptor Flk-1, angiopoetina e seu receptor Tie-2 e recertor tireoidiano (TRα) foram realizadas pela técnica de estreptavideina-biotina-peroxidade. A apoptose foi avaliada utilizando-se kit de TUNEL. A expressão relativa dos receptores de hormônios tireoidianos (TRα e TRß) foi avaliada pela técnica de RT-PCR tempo real. A expressão nuclear de CDC47 aumentou com a fase de maturação do oócito e foi observada nas células foliculares. Corpos apoptóticos foram observados nas células foliculares de folículos atrésicos e folículos pós-ovulatórios de ovários de peixes com 150g e 350g. A expressão de VEGF e do seu receptor Flk-1 foi também observada nas células foliculares , e a expressão de ambos aumentou com a maturidade do oócito , com uma maior intensidade no folículo maduro. A expressão de angiopoietina e do seu receptor (Tie 2) foi discreta e moderada, respectivamente. A expressão de TRα foi independente do desenvolvimento folicular. No entanto, a tilápia de 350g apresentou maior expressão de TRß em comparação com a tilápia de 50g. Conclui-se que a atividade proliferativa e a expressão de VEGF e de seu receptor aumenta com a maturação folicular e que a expressão dos TRs aumenta com a maturidade do ovário em tilápia (Oreochromis niloticus).(AU)
Assuntos
Animais , Feminino , Ovário , Receptores dos Hormônios Tireóideos , Apoptose , Neovascularização Fisiológica , Ciclídeos/anatomia & histologia , Imuno-Histoquímica/veterináriaResumo
A rare case of the co-occurrence of an osteogenesis imperfecta-like phenotype and congenital diaphragmatic hernia is reported in a male, mixed-breed kitten with a clinical history of dyspnea, dehydration, sternal recumbency and stupor. The animal presented moderate bone deformity of the fore and hind limbs, muscle atrophy, and cervical and thoracic lordosis. The radiological examination and necropsy revealed diffuse and intense radiolucency throughout the skeleton, curved or fractured bones, very thin cortical long bones, an intensely extended medullary canal and left diaphragmatic hernia with an aperture without bleeding or scarring. Microscopically, some long bones and vertebral bodies had less-differentiated cartilaginous epiphysis, predominantly attached to the epiphyseal plate and with absence of secondary ossification centers or incipient formation. The trabeculae were thin, few, surrounded by abundant cartilaginous tissue and coated with a layer of bulky cuboidal osteoblasts. The cortical long bones, vertebrae, skull and ribs were thin and discontinuous. Based on the clinical, radiological, macroscopic and microscopic findings, a diagnosis of osteogenesis imperfecta and congenital diaphragmatic hernia was confirmed. To the best of our knowledge, this report is the first case of OI associated with congenital diaphragmatic hernia in an animal.
Assuntos
Masculino , Animais , Gatos , Hérnias Diafragmáticas Congênitas/veterinária , Osteogênese Imperfeita/genética , Osteogênese Imperfeita/veterináriaResumo
In animals and humans, intestinal muscular hypertrophy has only been observed in the small intestine, and this appears to be the first report of the disease affecting the large intestine. A rare case of primary muscle hypertrophy of the colon was found during the necropsy of a male, 3-year-old, mixed breed cat that died due to poisoning by carbamate. The necropsy revealed that the entire colon had marked circumferential thickening with narrowing of the lumen. Microscopically, diffuse intense smooth muscle hypertrophy was observed, especially of the inner circular muscle layer and muscularis mucosa. Those finds were associated with moderate multifocal to coalescing lymphocytic inflammatory infiltrate. Smooth muscle fibers of the large and small intestine showed no CDC47 expression and the percentage of nuclei in the muscle layers was similar between the colon and the duodenum. Based on macroscopic, microscopic and immunohistochemical findings, the diagnosis of diffuse primary muscle hypertrophy of the colon was confirmed.
Assuntos
Animais , Gatos , Colo/patologia , Enteropatias/veterinária , Hipertrofia/veterinária , Autopsia/veterinária , Carbamatos/toxicidade , Colite/veterinária , Intoxicação/veterináriaResumo
In animals and humans, intestinal muscular hypertrophy has only been observed in the small intestine, and this appears to be the first report of the disease affecting the large intestine. A rare case of primary muscle hypertrophy of the colon was found during the necropsy of a male, 3-year-old, mixed breed cat that died due to poisoning by carbamate. The necropsy revealed that the entire colon had marked circumferential thickening with narrowing of the lumen. Microscopically, diffuse intense smooth muscle hypertrophy was observed, especially of the inner circular muscle layer and muscularis mucosa. Those finds were associated with moderate multifocal to coalescing lymphocytic inflammatory infiltrate. Smooth muscle fibers of the large and small intestine showed no CDC47 expression and the percentage of nuclei in the muscle layers was similar between the colon and the duodenum. Based on macroscopic, microscopic and immunohistochemical findings, the diagnosis of diffuse primary muscle hypertrophy of the colon was confirmed.(AU)
Assuntos
Animais , Gatos , Hipertrofia/veterinária , Enteropatias/veterinária , Colo/patologia , Carbamatos/toxicidade , Intoxicação/veterinária , Autopsia/veterinária , Colite/veterináriaResumo
Morphological and immunohistochemical characterization of angiogenic and apoptotic factors and the expression of thyroid receptors in the ovary of tilapia Oreochromis niloticus in captivity were studied. The morphological evaluation of the ovaries was performed by histological paraffin embedded and stained with HE. The immunohistochemical expressions of CDC47, VEGF, Flk-1, angiopoietin, Tie-2 and thyroid receptor (TRα) were performed by the technique of streptavidein-biotin-peroxidase. Apoptosis was assessed using the TUNEL kit. The relative expression of thyroid hormone receptors (TRα and TRβ) was assessed by RT-PCR real time. The nuclear expression of CDC47 increased with the stage of maturation of the oocyte and was observed in the follicle cells. Apoptotic bodies were observed in the follicular cells of atretic follicles and postovulatory follicles from the ovaries of 150g and 350g fish. Expression of VEGF and its receptor Flk-1 was also observed in the follicular cells, and the expression of both increased with the maturity of the oocyte, with a higher intensity observed in the full-grown follicle. The expression of angiopoietin and of its receptor (Tie 2) was discrete and moderate respectively. TRα expression was independent of follicular development. However, the 350 g tilapia exhibited higher expression of TRβ compared with the 50 g tilapia. We conclude that the proliferative activity and the expression of VEGF and its receptor increase with follicular maturation and that the TRs expression increases with ovarian maturity in tilapia (Oreochromis niloticus).(AU)
Foram estudadas as caracterizações morfológica e imuno-histoquímica de fatores angiogênicos e apoptóticos e a expressão de receptores tireoidianos no ovário de tilápia Oreochromis niloticus de cativeiro. A avaliação morfológica dos ovários foi realizada por cortes histológicos incluídos em parafina e corados por HE. As expressões imuno-histoquímicas de CDC47, VEGF e seu receptor Flk-1, angiopoetina e seu receptor Tie-2 e recertor tireoidiano (TRα) foram realizadas pela técnica de estreptavideina-biotina-peroxidade. A apoptose foi avaliada utilizando-se kit de TUNEL. A expressão relativa dos receptores de hormônios tireoidianos (TRα e TRβ) foi avaliada pela técnica de RT-PCR tempo real. A expressão nuclear de CDC47 aumentou com a fase de maturação do oócito e foi observada nas células foliculares. Corpos apoptóticos foram observados nas células foliculares de folículos atrésicos e folículos pós-ovulatórios de ovários de peixes com 150g e 350g. A expressão de VEGF e do seu receptor Flk-1 foi também observada nas células foliculares , e a expressão de ambos aumentou com a maturidade do oócito , com uma maior intensidade no folículo maduro. A expressão de angiopoietina e do seu receptor (Tie 2) foi discreta e moderada, respectivamente. A expressão de TRα foi independente do desenvolvimento folicular. No entanto, a tilápia de 350g apresentou maior expressão de TRβ em comparação com a tilápia de 50g. Conclui-se que a atividade proliferativa e a expressão de VEGF e de seu receptor aumenta com a maturação folicular e que a expressão dos TRs aumenta com a maturidade do ovário em tilápia (Oreochromis niloticus).(AU)