RESUMO
Background: Epithelial cell adhesion molecule (EpCAM) is a glycoprotein responsible for multiple cellular functions that includes cell-to-cell adhesion and signal transduction. Studies in human breast cancer demonstrated that its overexpression has been linked to a more aggressive histological phenotype. However, in canine mammary tumors, its use is still scarce and its function, remains unknown. The main goal of the present study was to characterize the expression of EpCAM in canine mammary malignant tumors and associate its expression with clinicopathological features. Materials, Methods & Results: Forty-eight canine mammary samples were analyzed in the present study and went through the Hematoxylin-Eosin (HE) routine technique for histopathological and immunohistochemical analysis. EpCAM expression was evaluated considering the total immunolabelling (TI) which consists of the addition of 2 parameters: proportion score (PS) and intensity score (IS). Thirteen (27.1%) cases showed weak expression, 9 (18.7%) were characterized by a moderate expression and 27 (54.2%) were classified as intense. EpCAM overexpression was described in 36 (72.9%) primary tumors and in 5 lymph node metastases (71.4%). Immunoreactivity was, mainly, characterized by a staining in the lateral cell membrane of luminal epithelial cells, both in primary tumors and lymph node metastases. The statistical analysis was performed using the SPSS Statistics, version 24. Clinicopathological features, such as histological grade (P = 0.027), intravascular emboli (P = 0.004) and lymph node metastases (P = 0.016) demonstrated to be associated with a more aggressive histological type. Intravascular emboli and lymph node metastases were also linked to a higher histological grade. Discussion: EpCAM overexpression has been described in the literature both in canine mammary tumors and breast cancer. While the results in breast cancer could be ambiguous, depending on the tumor subtype, in the present study our findings demonstrated that overexpression was present in more than 50% of the total cases evaluated. In addition, immunoreactivity was also in accordance with the literature that describes membranous staining as the most prevalent pattern type. It was not possible to correlate overexpression with histological type and histological grade. However, those clinicopathological features are essential to evaluate tumor prognosis. For instance, our results demonstrated that histological type was statistically significant with histological grade (P = 0.027), intravascular emboli (P = 0.004) and lymph node metastases (P = 0.016). Additionally, the Nottingham system was statistically significant with intravascular emboli (P = 0.033) and lymph node metastases (P = 0.006). These findings have also been described in similar studies using canine mammary tissues in bitches. When it comes to pattern of expression in primary tumors and lymph nodes metastases, our findings described that neoplastic cell presented the same phenotype in these 2 different locations. In addition, loss of expression was also described in cluster cells within the subcapsular area which reflects the idea that the expression of EpCAM molecule changes according to time, which could be translated into the multistep process of metastases. Finally, it demonstrates that the development of neoplasia is not something static, but rather dynamic.
Assuntos
Animais , Feminino , Cães , Neoplasias Mamárias Animais/química , Molécula de Adesão da Célula Epitelial/análise , Molécula de Adesão da Célula Epitelial/efeitos adversos , Imuno-Histoquímica/veterinária , HematoxilinaRESUMO
Background: Studies pointed out that the tumor-infiltrating lymphocytes (TILs) have considerable importance in caninemammary tumor (CMT). On the other hand, cancer cells sometimes find ways to use immune checkpoint proteins as ashield to avoid being identified and attacked by the immune system as programmed death 1 ligand 1 (PD-L1). In this study,it was investigated the relationship between PD-L1 expression, stromal tumor-infiltrating lymphocytes (TILs) in caninemammary tumor (CMT), and the association with clinical and pathological characteristics of the tumors.Materials, Methods & Results: PD-L1 expression and TILs were assessed in 23 female dogs with CMT. The tumors weregrouped into simple carcinoma (CA, n = 8) and complex carcinoma (CC, n = 15). Stromal TILs were assessed using twothresholds as TILs-Low representing < 50% of infiltrate within stromal area and TILs-High representing ≥ 50% of stromalarea. Clinicopathological data of CMT was characterized according to key parameters, as well as survival rates. TILs evaluation within tumor stroma revealed that 65.2% (n = 15) of tumors had TILs-Low. PD-L1 expression and stromal TILs weresignificantly associated (P = 0.009). PD-L1 expression was observed in 39% (n = 9) of all tumors of which 17.4% (n = 4)were from CA group and 21.7% (n = 5) were from CC group. PD-L1 expression within TILs was observed in 39% (n =9) of the tumors. PD-L1 in malignant epithelium was present in all lymph node metastasis (n = 5). PD-L1 was associatedwith involvement of regional lymph nodes (P = 0.034). Survival curves demonstrated TILs-Low had higher (P = 0.010)overall survival (OS) compared with TILs-High, and PD-L1+ and PD-L1(P = 0.06) did not differed. The clinicopathological variables significantly correlated with OS by univariate analysis were the histological grade (P = 0.009), lymphnode involvement (P = 0.004), stromal...(AU)
Assuntos
Animais , Feminino , Cães , Cães , Neoplasias da Mama/veterinária , Linfócitos , Proteínas de Membrana , Mastectomia Segmentar/veterináriaRESUMO
We investigated the applicability of ssGBLUP methodology under the autoregressive model (H-AR) for genomic evaluation of longitudinal reproductive traits in Portuguese Holstein cattle. The genotype data of 1,230 bulls and 1,645 cows were considered in our study. The reproductive traits evaluated were interval from calving to first service (ICF), calving interval (CI) and daughter pregnancy rate (DPR) measured during the first four parities. Reliability and rank correlation were used to compare the H-AR with the traditional pedigree-based autoregressive models (A-AR). In addition, a validation study was performed considering different scenarios. Higher genomic estimated breeding values (GEBV) reliabilities were obtained for genotyped bulls when evaluated under the H-AR model, with emphasis on bulls with less than 9 daughters. For this group, the averages of GEBV reliabilities corresponded to 0.62, 0.69 and 0.62 for ICF, CI and DPR, respectively, while the averages obtained by the A-AR model were 0.27, 0.15 and 0.16. The validation study was favourable to H-AR. The best results were observed in the scenario where genotyped cows were combined with contributing bulls (genotyped bulls with daughter or relationship information in the population). Overall, the results suggest that ssGBLUP methodology under the autoregressive model is a feasible and applicable approach to be used in genomic analyses of longitudinal reproductive traits in Portuguese Holstein cattle.
Assuntos
Genoma , Animais , Bovinos , Feminino , Genômica , Genótipo , Masculino , Modelos Genéticos , Linhagem , Fenótipo , Portugal , Gravidez , Reprodutibilidade dos TestesRESUMO
Reproductive efficiency is major determinant of the dairy herd profitability. Thus, reproductive traits have been widely used as selection objectives in the current dairy cattle breeding programs. We aimed to evaluate strategies to model days open (DO), calving interval (CI) and daughter pregnancy rate (DPR) in Brazilian Holstein cattle. These reproductive traits were analysed by the autoregressive (AR) model and compared with classical repeatability (REP) model using 127,280, 173,092 and 127,280 phenotypic records, respectively. The first three calving orders of cows from 1,469 Holstein herds were used here. The AR model reported lower values for Akaike Information Criteria and Mean Square Errors, as well as larger model probabilities, for all evaluated traits. Similarly, larger additive genetic and lower residual variances were estimated from AR model. Heritability and repeatability estimates were similar for both models. Heritabilities for DO, CI and DPR were 0.04, 0.07 and 0.04; and 0.05, 0.06 and 0.04 for AR and REP models, respectively. Individual EBV reliabilities estimated from AR for DO, CI and DPR were, in average, 0.29, 0.30 and 0.29 units higher than those obtained from REP model. Rank correlation between EBVs obtained from AR and REP models considering the top 10 bulls ranged from 0.72 to 0.76; and increased from 0.98 to 0.99 for the top 100 bulls. The percentage of coincidence between selected bulls from both methods increased over the number of bulls included in the top groups. Overall, the results of model-fitting criteria, genetic parameters estimates and EBV predictions were favourable to the AR model, indicating that it may be applied for genetic evaluation of longitudinal reproductive traits in Brazilian Holstein cattle.
Assuntos
Bovinos/genética , Fertilidade/genética , Reprodução/genética , Animais , Brasil , Cruzamento , Bovinos/fisiologia , Indústria de Laticínios/métodos , Feminino , Masculino , Modelos Estatísticos , Gravidez , Taxa de GravidezRESUMO
Background: Studies pointed out that the tumor-infiltrating lymphocytes (TILs) have considerable importance in caninemammary tumor (CMT). On the other hand, cancer cells sometimes find ways to use immune checkpoint proteins as ashield to avoid being identified and attacked by the immune system as programmed death 1 ligand 1 (PD-L1). In this study,it was investigated the relationship between PD-L1 expression, stromal tumor-infiltrating lymphocytes (TILs) in caninemammary tumor (CMT), and the association with clinical and pathological characteristics of the tumors.Materials, Methods & Results: PD-L1 expression and TILs were assessed in 23 female dogs with CMT. The tumors weregrouped into simple carcinoma (CA, n = 8) and complex carcinoma (CC, n = 15). Stromal TILs were assessed using twothresholds as TILs-Low representing < 50% of infiltrate within stromal area and TILs-High representing ≥ 50% of stromalarea. Clinicopathological data of CMT was characterized according to key parameters, as well as survival rates. TILs evaluation within tumor stroma revealed that 65.2% (n = 15) of tumors had TILs-Low. PD-L1 expression and stromal TILs weresignificantly associated (P = 0.009). PD-L1 expression was observed in 39% (n = 9) of all tumors of which 17.4% (n = 4)were from CA group and 21.7% (n = 5) were from CC group. PD-L1 expression within TILs was observed in 39% (n =9) of the tumors. PD-L1 in malignant epithelium was present in all lymph node metastasis (n = 5). PD-L1 was associatedwith involvement of regional lymph nodes (P = 0.034). Survival curves demonstrated TILs-Low had higher (P = 0.010)overall survival (OS) compared with TILs-High, and PD-L1+ and PD-L1 (P = 0.06) did not differed. The clinicopathological variables significantly correlated with OS by univariate analysis were the histological grade (P = 0.009), lymphnode involvement (P = 0.004), stromal...
Assuntos
Feminino , Animais , Cães , Cães , Linfócitos , Neoplasias da Mama/veterinária , Proteínas de Membrana , Mastectomia Segmentar/veterináriaRESUMO
This study focused on the identification of QTL regions, candidate genes, and network related genes based on the first 3 lactations (LAC3) of milk, fat, and protein yields, and somatic cell score (SCS) in Portuguese Holstein cattle. Additionally, the results were compared with those from only first lactation (LAC1) data. The analyses were performed using the weighted single-step GWAS under an autoregressive test-day (TD) multiple lactations model. A total of 11,434,294 and 4,725,673 TD records from LAC3 and LAC1, respectively, including 38,323 autosomal SNPs and 1338 genotyped animals were used in GWAS analyses. A total of 51 (milk), 5 (fat), 24 (protein), and 4 (SCS) genes were associated to previously annotated relevant QTL regions for LAC3. The CACNA2D1 at BTA4 explained the highest proportion of genetic variance respectively for milk, fat, and protein yields. For SCS, the TRNAG-CCC at BTA14, MAPK10, and PTPN3 genes, both at BTA6 were considered important candidate genes. The accessed network refined the importance of the reported genes. CACNA2D1 regulates calcium density and activation/inactivation kinetics of calcium transport in the mammary gland; whereas TRNAG-CCC, MAPK10, and PTPN3 are directly involved with inflammatory processes widely derived from mastitis. In conclusion, potential candidate genes (TRNAG-CCC, MAPK10, and PTPN3) associated with somatic cell were highlighted, which further validation studies are needed to clarify its mechanism action in response to mastitis. Moreover, most of the candidate genes identified were present in both (LAC3 and LAC1) for milk, fat and protein yields, except for SCS, in which no candidate genes were shared between LAC3 and LAC1. The larger phenotypic information provided by LAC3 dataset was more effective to identify relevant genes, providing a better understanding of the genetic architecture of these traits over all lactations simultaneously.
Assuntos
Redes Reguladoras de Genes , Estudos de Associação Genética/veterinária , Lactação/genética , Leite , Locos de Características Quantitativas , Animais , Bovinos , Feminino , Genótipo , Modelos Genéticos , Fenótipo , Polimorfismo de Nucleotídeo Único , PortugalRESUMO
We investigated the efficiency of the autoregressive repeatability model (AR) for genetic evaluation of longitudinal reproductive traits in Portuguese Holstein cattle and compared the results with those from the conventional repeatability model (REP). The data set comprised records taken during the first four calving orders, corresponding to a total of 416, 766, 872 and 766 thousand records for interval between calving to first service, days open, calving interval and daughter pregnancy rate, respectively. Both models included fixed (month and age classes associated to each calving order) and random (herd-year-season, animal and permanent environmental) effects. For AR model, a first-order autoregressive (co)variance structure was fitted for the herd-year-season and permanent environmental effects. The AR outperformed the REP model, with lower Akaike Information Criteria, lower Mean Square Error and Akaike Weights close to unity. Rank correlations between estimated breeding values (EBV) with AR and REP models ranged from 0.95 to 0.97 for all studied reproductive traits, when the total bulls were considered. When considering only the top-100 selected bulls, the rank correlation ranged from 0.72 to 0.88. These results indicate that the re-ranking observed at the top level will provide more opportunities for selecting the best bulls. The EBV reliabilities provided by AR model was larger for all traits, but the magnitudes of the annual genetic progress were similar between two models. Overall, the proposed AR model was suitable for genetic evaluations of longitudinal reproductive traits in dairy cattle, outperforming the REP model.
Assuntos
Bovinos/genética , Reprodução/genética , Animais , Cruzamento/métodos , Bovinos/fisiologia , Indústria de Laticínios/métodos , Feminino , Modelos Genéticos , Gravidez , Característica Quantitativa HerdávelRESUMO
Autoregressive (AR) and random regression (RR) models were fitted to test-day records from the first three lactations of Brazilian Holstein cattle with the objective of comparing their efficiency for national genetic evaluations. The data comprised 4,142,740 records of milk yield (MY) and somatic cell score (SCS) from 274,335 cows belonging to 2,322 herds. Although heritabilities were similar between models and traits, additive genetic variance estimates using AR were 7.0 (MY) and 22.2% (SCS) higher than those obtained from RR model. On the other hand, residual variances were lower in both traits when estimated through AR model. The rank correlation between EBV obtained from AR and RR models was 0.96 and 0.94 (MY) and 0.97 and 0.95 (SCS), respectively, for bulls (with 10 or more daughters) and cows. Estimated annual genetic gains for bulls (cows) obtained using AR were 46.11 (49.50) kg for MY and -0.019 (-0.025) score for SCS; whereas using RR these values were 47.70 (55.56) kg and -0.022 (-0.028) score. Akaike information criterion was lower for AR in both traits. Although AR model is more parsimonious, RR model assumes genetic correlations different from the unity within and across lactations. Thus, when these correlations are relatively high, these models tend to yield to similar predictions; otherwise, they will differ more and RR model would be theoretically sounder.