RESUMO
This study was undertaken with the aim to evaluate the p53 expression, applying the immunohistochemical technique to malignant primary mammary neoplasms histopathologically diagnosed in female dogs, and to investigate exon 8 of the Tp53 suppressor gene for mutation types by means of PCR-RFLP pattern of bands. Nineteen healthy mammary glands were used as a control group (group 1). Samples from 18 cases diagnosed with malignant mammary tumors (group 2), and the contralateral mammary glands (group 3) were collected during the UFRPE Veterinary Hospital routine. The tumors were diagnosed by histopathology and subdivided into grades of malignity. The streptavidin-biotin peroxidase method was used to analyze the immunohistochemical expression of p53, evaluated according to the location and intensity of stain. Expression of p53 protein was not observed in the samples of group 1. On the contrary, it was observed in all malignant tumors; the protein p53 was localized either only in the nucleus or in the nucleus and in the cytoplasm, in samples of group 2. In group 3, expression of p53 protein was observed in the tumors (2 samples) and in normal mammary tissues (4 samples). For the molecular analyses, genomic DNA was extracted and submitted to PCR-RFLP with the following endonuclease enzymes: AluI, BsoBI, DdeI and SmaI. The band pattern showed polymorphism between groups, but not between histological variants of tumors. This polymorphism detected mutations in the fragment studied - exon 8 of Tp53 - which could account for changes in nucleotides, located in the restriction sites of the endonuclease enzymes. In conclusion, the immunoexpression of p53 had no relationship with histological subtype or malignity grade, but it can be related to the presence of mammary tumors in female dogs. The PCR-RFLP technique can be an important tool for the study of mammary carcinogenesis in bitches because the polymorphism obtained may allow early diagnosis in tissues of mammary glands.
Este estudo foi realizado com o objetivo de avaliar a expressão da proteína p53, pela técnica de imuno-histoquímica, em neoplasmas mamários malignos em cadelas, além de investigar mutações no éxon 8 do gene supressor Tp53 por meio do padrão de bandas obtidas por PCR-RFLP. Dezenove mamas de cadelas saudáveis foram usadas como controle (Grupo 1). Amostras de 18 casos de tumores malignos (Grupo 2) e suas glândulas mamárias contralaterais (Grupo 3) foram obtidas na rotina do Hospital Veterinário da UFRPE. Os tumores foram identificados histologicamente e classificados em graus de malignidade. O método da estreptoavidina-biotina peroxidase foi utilizado para a análise da expressão de p53 por imuno-histoquímica, de acordo com a localização e intensidade da coloração. A expressão da proteína p53 não foi observada nas amostras do Grupo 1, mas foi encontrada em todas as amostras de tumores malignos (Grupo 2) seja só no núcleo, ou também no citoplasma. No Grupo 3, a expressão foi observada em quatro amostras normais e em duas que apresentavam tumor. Para a análise molecular, o DNA genômico foi extraído e submetido à PCR-RFLP com as seguintes endonucleases: AluI, BsoBI, DdeI e SmaI. O padrão de bandas foi polimórfico entre os grupos, mas não entre as variantes tumorais. Esse polimorfismo detectou mutações no fragmento estudado - éxon 8 do gene Tp53 - que podem resultar em alterações nos nucleotídeos, localizados nos sítios de restrição das enzimas. Esses achados levam a conclusão de que a imunoexpressão da p53 não tem relação com o subtipo histológico ou grau de malignidade do tumor, mas sim com a presença dos tumores no tecido mamário de cadelas. A PCR-RFLP pode ser usada como importante ferramenta para o estudo da carcinogênese mamária na cadela, possibilitando gerar diagnósticos precoces através do polimorfismo obtido com endonucleases de restrição pré-selecionadas.
Assuntos
Animais , Feminino , Cães , Neoplasias da Mama/veterinária , Polimorfismo de Fragmento de RestriçãoRESUMO
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.
RESUMO
This study was undertaken with the aim to evaluate the p53 expression, applying the immunohistochemical technique to malignant primary mammary neoplasms histopathologically diagnosed in female dogs, and to investigate exon 8 of the Tp53 suppressor gene for mutation types by means of PCR-RFLP pattern of bands. Nineteen healthy mammary glands were used as a control group (group 1). Samples from 18 cases diagnosed with malignant mammary tumors (group 2), and the contralateral mammary glands (group 3) were collected during the UFRPE Veterinary Hospital routine. The tumors were diagnosed by histopathology and subdivided into grades of malignity. The streptavidin-biotin peroxidase method was used to analyze the immunohistochemical expression of p53, evaluated according to the location and intensity of stain. Expression of p53 protein was not observed in the samples of group 1. On the contrary, it was observed in all malignant tumors; the protein p53 was localized either only in the nucleus or in the nucleus and in the cytoplasm, in samples of group 2. In group 3, expression of p53 protein was observed in the tumors (2 samples) and in normal mammary tissues (4 samples). For the molecular analyses, genomic DNA was extracted and submitted to PCR-RFLP with the following endonuclease enzymes: AluI, BsoBI, DdeI and SmaI. The band pattern showed polymorphism between groups, but not between histological variants of tumors. This polymorphism detected mutations in the fragment studied - exon 8 of Tp53 - which could account for changes in nucleotides, located in the restriction sites of the endonuclease enzymes. In conclusion, the immunoexpression of p53 had no relationship with histological subtype or malignity grade, but it can be related to the presence of mammary tumors in female dogs. The PCR-RFLP technique can be an important tool for the study of mammary carcinogenesis in bitches because the polymorphism obtained may allow early diagnosis in tissues of mammary glands.(AU)
Este estudo foi realizado com o objetivo de avaliar a expressão da proteína p53, pela técnica de imuno-histoquímica, em neoplasmas mamários malignos em cadelas, além de investigar mutações no éxon 8 do gene supressor Tp53 por meio do padrão de bandas obtidas por PCR-RFLP. Dezenove mamas de cadelas saudáveis foram usadas como controle (Grupo 1). Amostras de 18 casos de tumores malignos (Grupo 2) e suas glândulas mamárias contralaterais (Grupo 3) foram obtidas na rotina do Hospital Veterinário da UFRPE. Os tumores foram identificados histologicamente e classificados em graus de malignidade. O método da estreptoavidina-biotina peroxidase foi utilizado para a análise da expressão de p53 por imuno-histoquímica, de acordo com a localização e intensidade da coloração. A expressão da proteína p53 não foi observada nas amostras do Grupo 1, mas foi encontrada em todas as amostras de tumores malignos (Grupo 2) seja só no núcleo, ou também no citoplasma. No Grupo 3, a expressão foi observada em quatro amostras normais e em duas que apresentavam tumor. Para a análise molecular, o DNA genômico foi extraído e submetido à PCR-RFLP com as seguintes endonucleases: AluI, BsoBI, DdeI e SmaI. O padrão de bandas foi polimórfico entre os grupos, mas não entre as variantes tumorais. Esse polimorfismo detectou mutações no fragmento estudado - éxon 8 do gene Tp53 - que podem resultar em alterações nos nucleotídeos, localizados nos sítios de restrição das enzimas. Esses achados levam a conclusão de que a imunoexpressão da p53 não tem relação com o subtipo histológico ou grau de malignidade do tumor, mas sim com a presença dos tumores no tecido mamário de cadelas. A PCR-RFLP pode ser usada como importante ferramenta para o estudo da carcinogênese mamária na cadela, possibilitando gerar diagnósticos precoces através do polimorfismo obtido com endonucleases de restrição pré-selecionadas.(AU)
Assuntos
Animais , Feminino , Cães , Polimorfismo de Fragmento de RestriçãoRESUMO
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 +-g d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 +- 1.1h vs. 13.8 +- 2.6h vs. 13.3 +-1.4h), as well as estrus duration (33.6 +- 7.3h vs. 29.6 +-3.2h vs. 32.8 +- 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.
Objetivou-se avaliar o efeito da utilização do mesmo dispositivo de liberação controlada de drogas (CIDR) por até três vezes sobre o desempenho reprodutivo de cabras leiterias exploradas no semiárido do Nordeste Brasileiro. Foram utilizadas 45 cabras divididas em três tratamentos de sincronização do estro, sendo: CIDR1x, tratadas com CIDR novo durante nove dias. Dois dias antes da retirada do dispositivo, foram aplicados 75 +-g de d-cloprostenol e 300 UI de gonadotrofina coriônica eqüina (eCG). Para os demais tratamentos, foi utilizado o mesmo protocolo hormonal, diferindo apenas pelo uso do mesmo CIDR pela segunda vez no grupo CIDR2x e uso pela terceira vez no grupo CIDR3x. O intervalo entre a retirada do dispositivo e o início do estro (13,3+- 1,1h vs. 13,8 +- 2,6h vs. 13,3 +- 1,4h), bem como, a duração do estro (33,6 +- 7,3h vs. 29,6 +- 3,2h vs. 32,8 +- 4,5h) não diferiram (p > 0,05) entre os grupos CIDR1x, CIDR2x e CIDR3x, respectivamente. Todas as fêmeas sincronizadas foram identificadas em estro. As médias de fertilidade e prolificidade média após inseminação artificial foram, respectivamente, de 82,2% e 1,9 crias, não havendo diferença significativa (p > 0,05) entre os tratamentos. A utilização do mesmo CIDR por até três vezes foi viável na sincronização do estro de caprinos leiteiros.
Assuntos
Animais , Cabras , Fertilidade , Liberação Controlada de Fármacos , HormôniosRESUMO
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.
RESUMO
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.