Search

The Himalayan plant Inula racemosa has medicinal properties and can be used to prevent or treat cancer. This is because it contains certain chemicals that are known to fight cancer cells with few or no side effects. I. racemosa has been used for this purpose for many years in traditional medicine and has shown promising results. The present study was crafted to explore the suppressive impacts on cellular proliferation of the root extract derived from I. racemosa via in vivo experimentation. I. racemosa (IR) root extract was tested at three different doses (100, 250, and 500 mg/Kg BW) for 18 weeks to assess its anti‐neoplastic activity against mammary tumors in female rats. The assessment included various parameters such as hematological and biochemical indices, tumor parameters, oxidative stress analysis, gross and histopathological lesion determination, Masson's trichrome staining, immunohistochemical expression of Ki‐67, MMP‐9, and VEGF in mammary gland tissues, and molecular docking. The chemopreventive action of IR root extract was demonstrated by the inhibition of tumor parameters (tumor size and tumor volume), minimum changes in the liver (ALT, AST, and ALP) and kidney enzymes (BUN and creatinine), declined lipid peroxidation activity, decline gross, and histological changes in mammary gland tumors, reduced expression of KI‐67, MMP‐9, and VEGF and maximum binding affinity of isoalantolactone with VEGF through molecular docking. The study suggests that the active constituents (isoalantolactone and alantolactone) of I. racemosa roots have anti‐neoplastic activity against mammary tumors, making them a valuable therapeutic regimen for the future. [ABSTRACT FROM AUTHOR]

Canine mammary tumors (CMTs) represent a significant health concern in dogs, with a high incidence among intact female dogs. CMTs are a promising comparative model for human breast cancer, due to sharing several pathophysiological features. Additionally, CMTs have a strong genetic correlation with their human counterpart, including the expression of microRNAs (miRNAs). MiRNAs are a class of non-coding RNAs that play important roles in post-translational regulation of gene expression, being implicated in carcinogenesis, tumor progression, and metastasis. Moreover, miRNAs hold promise as diagnostic, prognostic, and metastatic biomarkers. Understanding the molecular mechanisms underlying CMTs is crucial for improving diagnosis, prognosis, and monitoring of treatments. Herein, we provide a comprehensive overview of the current knowledge on miRNAs in CMTs, highlighting their roles in carcinogenesis and their potential as biomarkers. Additionally, we highlight the current limitations and critically discuss the overarching challenges in this field, emphasizing the need for future research to translate miRNA findings into veterinary clinical practice. [ABSTRACT FROM AUTHOR]

Aging is a pivotal risk factor for cancer, yet the underlying mechanisms remain poorly defined. Here, we explore age-related changes in the rat mammary gland by single-cell multiomics. Our findings include increased epithelial proliferation, loss of luminal identity, and decreased naive B and T cells with age. We discover a luminal progenitor population unique to old rats with profiles reflecting precancerous changes and identify midkine (Mdk) as a gene upregulated with age and a regulator of age-related luminal progenitors. Midkine treatment of young rats mimics age-related changes via activating PI3K-AKT-SREBF1 pathway and promotes nitroso-N-methylurea-induced mammary tumorigenesis. Midkine levels increase with age in human blood and mammary epithelium, and higher MDK in normal breast tissue is associated with higher breast cancer risk in younger women. Our findings reveal a link between aging and susceptibility to tumor initiation and identify midkine as a mediator of age-dependent increase in breast tumorigenesis. [Display omitted] • Cellular and molecular changes in the mammary gland with age • Midkine is a key driver of aging-related changes and mammary tumorigenesis • High midkine levels are associated with higher risk of breast cancer in younger women Yan et al. reveal age-related changes in the rat mammary gland, identifying a unique aging-associated luminal progenitor population. Midkine treatment mimics the effects of aging and promotes mammary tumorigenesis. Elevated midkine levels are associated with increased breast cancer risk in younger women highlighting its role in age-related tumor initiation. [ABSTRACT FROM AUTHOR]

Nowadays, the use of food additives, such as Sunset Yellow (SY), is growing, which attracted attention to the potential relationship between some diseases and food additives. The study aimed to investigate the role of Sunset Yellow during chemically-induced mammary gland carcinogenesis in Sprague–Dawley rats. Three groups of female rats were intraperitoneally administered with N-methyl-N-nitrosourea (MNU). Group 1 was set on a basal diet. Group 2 was treated with 161.4 mg\kg\day Sunset Yellow (SY). Group 3 was given SY at 80.7 mg\kg\day. Groups 4–6 were not administered MNU; Group 4 received vehicles only. Groups 5 and 6 were administered SY similarly to groups 2 and 3 respectively. Sunset Yellow at both doses exerted a significant dose-dependent increase in tumor incidences, multiplicities, volumes, and decreased tumor latency as compared with control. Immunolabeling indexes of the proliferating cell nuclear antigen, estrogen receptor alpha, and progesterone receptor were significantly increased after SY treatment. Oxidative stress markers, serum estrogen, progesterone, and prolactin levels were significantly modified by SY treatment. The mRNA expression of estrogen receptor alpha and epidermal growth factor was up-regulated in SY groups versus control. Collectively, SY has significantly promoted MNU-induced mammary tumors in rats with underlying mechanisms correlating SY consumption with estrogen disruption and subsequent antioxidative stress discrepancy. [Display omitted] • Sunset Yellow (SY) is a food coloring dye. • SY has significantly promoted MNU-induced mammary tumors in rats. • SY exaggerated tumor growth via estrogen disruption and subsequent antioxidative stress discrepancy. [ABSTRACT FROM AUTHOR]

ABSTRACT: Lymph node status is considered an important clinical prognostic factor in canine mammary carcinomas and women’s breast neoplasms. However, occult isolated tumor cells (ITCs) can be missed during hematoxylin and eosin (HE) analyses. Immunohistochemistry (IHC) for cytokeratin can be used to detect carcinomatous occult ITCs in mammary drainage lymph nodes. However, brown pigments, such as hemosiderin and ceroid in lymph nodes, may hinder the search for occult metastases by IHC utilizing DAB (3,3′-diaminobenzidine) as the chromogen. The aim of this study was to identify ITCs in canine lymph nodes of cases in which it was not detectable by routine HE evaluation through IHC for cytokeratin (AE1/AE3) combined with histochemistry techniques, such as Perls’ Prussian blue and periodic acid-Schiff (PAS), to improve the detection of occult metastases when hemosiderin and ceroid were present in these lymph nodes. For this, 25 tubulopapillary mammary carcinomas with their respective submitted 29 regional lymph nodes, previously given as free of tumor cells by HE analyses, were selected. Mammary tumors were graduated, and vascular invasion was investigated in these tumors. The submitted lymph nodes were reevaluated in HE, looking for occult metastases. IHC for cytokeratin (AE1/AE3) was used to detect occult metastases in mammary lymph nodes. Subsequently, a combined technique of IHC with Perl’s Prussian blue (for hemosiderin) or PAS (for ceroid) was performed to optimize the detection of ITCs by IHC, distinguishing them from pigments. Occult metastases were classified by their microanatomical location in subcapsular, cortical and medullary. Hemosiderin and ceroid were searched in lymph nodes and quantified as low, moderate, or high. The amount of pigments with a percentage of ITCs was also compared. Isolated tumor cells were found in 24.1% (7/29) of mammary lymph nodes. These ITCs were located mainly in subcapsular sinuses (4/7; 57.1%), followed by cortical (2/7; 28.5%) and medullary sinuses (1/7; 14.3%). There were concomitant lymph nodes with ITCs in 33.4% (2/6) of cases with vascular invasion. Hemosiderin and ceroid were present in about 90% of the 29 lymph nodes analyzed. In 42.8% (3/7) of lymph nodes with ITCs, hemosiderin and/or ceroid were in the same location as ITCs. It was found that lymph nodes in which ITCs were detected also present high amounts of hemosiderin (3/7; 42.9%) and low amounts of ceroid (5/7; 71.4%). In this study, IHC for cytokeratin (AE1/AE3) was an efficient method to detect occult tumor cells. IHC combined with Perls’ Prussian blue or with PAS proved to be a helpful way to investigate the presence of occult metastases in the lymph nodes of mammary canine tumors. It allowed distinguishing hemosiderin and ceroid, respectively, from ITCs in the same slide of IHC (immunostained by DAB), favoring a more accurate analysis by pathologists, which can be useful for the oncological staging of these patients.

Purpose: Alcohol consumption increases the risk of breast cancer and promotes cancer progression. Alcohol exposure could affect both processes of the mammary carcinogenesis, namely, the cell transformation and onset of tumorigenesis as well as cancer aggressiveness including metastasis and drug resistance/recurrence. However, the cellular and molecular mechanisms underlying alcohol tumor promotion remain unclear. There are four members of the mammalian p38 mitogen-activated protein kinase (MAPK) family, namely, p38α, p38β, p38γ and p38δ. We have previously demonstrated alcohol exposure selectively activated p38γ MAPK in breast cancer cells in vitro and in vivo. Pirfenidone (PFD), an antifibrotic compound approved for the treatment of idiopathic pulmonary fibrosis, is also a pharmacological inhibitor of p38γ MAPK. This study aimed to determine whether PFD is useful to inhibit alcohol-induced promotion of breast cancer. Methods: Female adolescent (5 weeks) MMTV-Wnt1 mice were exposed to alcohol with a liquid diet containing 6.7% ethanol. Some mice received intraperitoneal (IP) injection of PFD (100 mg/kg) every other day. After that, the effects of alcohol and PFD on mammary tumorigenesis and metastasis were examined. Results: Alcohol promoted the progression of mammary tumors in adolescent MMTV-Wnt1 mice. Treatment of PFD blocked tumor growth and alcohol-promoted metastasis. It also significantly inhibited alcohol-induced tumorsphere formation and cancer stem cell (CSC) population. Conclusion: PFD inhibited mammary tumor growth and alcohol-promoted metastasis. Since PFD is an FDA-approved drug, the current findings may be helpful to re-purpose its application in treating aggressive breast cancer and alcohol-promoted mammary tumor progression. [ABSTRACT FROM AUTHOR]

A particular type of tumor that is frequently detected in female dogs who are sexually active is a mammary tumor. Neoplasia results from DNA-based alterations in cell cycle regulating genes. The mammary gland is prone to the formation of tumors due to its dynamic structure. The development of this tumor is supported by numerous variables. It has been recently discovered that there is substantial evidence linking the BRCA2 gene to the process of cancer. Standard examination techniques, such as fine needle aspiration, histopathology, and immunohistochemistry, are used along with ancillary tests to determine the tumor type and degree of malignancy. The primary treatment option for malignant tumors is surgical resection followed by adjuvant chemotherapy; benign tumors necessitate surgical resection as well. Adjuvant therapy options include hormone therapy and non-steroidal anti-inflammatory medications. Tumor tissue undergoes angiogenesis as it grows and develops to accommodate the abundant supply of nutrients. Therefore, angiogenesis-inhibiting therapies can be utilized to halt the growth of tumor cells. A number of antiangiogenic medications are now being studied in clinical settings on humans, and several more are undergoing trials on animals. In addition to pharmaceuticals, viruses may be used as a therapeutic to block tumor angiogenesis.

Background: MMTV causes mammary tumors in mice, and it is associated with invasive and aggressive forms of breast cancer in humans. However, the underlying mechanisms are yet unknown. We aimed to determine the MMTV-like virus (MMTV-LV) association with histological types of breast cancer, nodal involvement, and metastasis. Methods: First, 105 breast cancer biopsies and 15 disease-free biopsies were collected. Details of clinicopathological characteristics were retrieved from patients’ records. The status of MMTV-LV was already known for these biopsy samples. Associations of MMTV-LV prevalence with LNM status and metastatic history were determined. Next, quantitative PCR (qPCR) was used to quantify env gene mRNA in biopsies positive for MMTV-LV. Expression of the env gene was compared against different histopathological types of mammary tumor, LNM status, and metastasis by performing Ordinary One Way ANOVA followed by Tukey’s multiple comparisons test. Results: MMTV-LV prevalence was found to have no significant association with LNM or metastatic history. As compared to normal control, expression of the env gene was significantly higher (>2.8 folds) in invasive samples (P-value: < 0.01). Expression was also higher (3.28 and 2.89 folds) in patient samples with LNM (P-value: 0.0006) or metastatic history (P-value: < 0.0001), respectively. Conclusion: We conclude that MMTV-LV prevalence is not associated with LNM status or breast cancer metastasis; samples with invasive phenotypes, nodal involvement, and metastasis exhibit significantly higher expression of the MMTV-like env gene.

The heterogeneous nature of human breast cancer (HBC) can still lead to therapy inefficacy and high lethality, and new therapeutics as well as new spontaneous animal models are needed to benefit translational HBC research. Dogs are primarily investigated since they spontaneously develop tumors that share many features with human cancers. In recent years, different natural phytochemicals including berberine, a plant alkaloid, have been reported to have antiproliferative activity in vitro in human cancers and rodent animal models. In this study, we report the antiproliferative activity and mechanism of action of berberine, its active metabolite berberrubine, and eight analogs, on a canine mammary carcinoma cell line and in transgenic zebrafish models. We demonstrate both in vitro and in vivo the significant effects of specific analogs on cell viability via the induction of apoptosis, also identifying their role in inhibiting the Wnt/β-catenin pathway and activating the Hippo signals with a downstream reduction in CTGF expression. In particular, the berberine analogs NAX035 and NAX057 show the highest therapeutic efficacy, deserving further analyses to elucidate their mechanism of action more in detail, and in vivo studies on spontaneous neoplastic diseases are needed, aiming at improving veterinary treatments of cancer as well as translational cancer research. [ABSTRACT FROM AUTHOR]

Simple Summary: Due to the limited options in mammary tumor treatments, the following study demonstrates the efficiency of Doxorubicin on canine mammary tumor therapy and in the alteration of expression of the key genes involved in the epithelial-to-mesenchymal transition processes. Canine mammary tumors represent one of the leading malignant pathologies in female dogs, displaying the importance of efficient therapeutic findings, besides the golden-standard surgery, able to limit the development of the disease. Studies in human cancers demonstrated that Doxorubicin presents a good effect in different biological processes like apoptosis, autophagy, the cell cycle, cell invasion, and the epithelial-to-mesenchymal transition. This study followed the effects of Doxorubicin on two canine mammary cancer cell lines P114 and CMT-U27. Doxorubicin treatment in both cell lines shows an inhibitory effect in cell proliferation and an alteration in expression of the EMT-related genes. The obtained results provide valuable information for revealing the link between Doxorubicin, phenotypic changes, and proliferation dynamics in canine mammary tumor models. [ABSTRACT FROM AUTHOR]

A particular type of tumor that is frequently detected in female dogs who are sexually active is a mammary tumor. Neoplasia results from DNA-based alterations in cell cycle regulating genes. The mammary gland is prone to the formation of tumors due to its dynamic structure. The development of this tumor is supported by numerous variables. It has been recently discovered that there is substantial evidence linking the BRCA2 gene to the process of cancer. Standard examination techniques, such as fine needle aspiration, histopathology, and immunohistochemistry, are used along with ancillary tests to determine the tumor type and degree of malignancy. The primary treatment option for malignant tumors is surgical resection followed by adjuvant chemotherapy; benign tumors necessitate surgical resection as well. Adjuvant therapy options include hormone therapy and non-steroidal anti-inflammatory medications. Tumor tissue undergoes angiogenesis as it grows and develops to accommodate the abundant supply of nutrients. Therefore, angiogenesis-inhibiting therapies can be utilized to halt the growth of tumor cells. A number of antiangiogenic medications are now being studied in clinical settings on humans, and several more are undergoing trials on animals. In addition to pharmaceuticals, viruses may be used as a therapeutic to block tumor angiogenesis. [ABSTRACT FROM AUTHOR]

Simple Summary: The increasing incidence of cancer in animals is a growing concern, with mammary cancer being the most prevalent form in female dogs. Efforts have been made to prevent and understand the factors contributing to its development. Using data from a veterinary hospital of reference in Brazil, we determined that the size dog, breed type, housing, and body score influence the risk of developing malignant mammary tumors. Mammary cancer is one of the main causes of death in female dogs worldwide, considering that many risk factors are involved in its development. This study aimed to elucidate the relationship between epidemiological and clinical risk factors with the histopathological diagnosis of malignant mammary tumors in dogs treated at the Veterinary Hospital of the Federal University of Uberlândia, which has one of the first veterinary oncology services in Brazil. A retrospective matched case-control study was conducted to identify risk factors for the development of malignant mammary tumors in dogs. The variables analyzed were size dog, breed, housing, type of diet, and body score. Potential risk factors were selected by univariate analysis (p < 0.25) before multivariate forward binary logistic regression. The most frequent benign tumor was the benign mixed tumor (35.2%), and the most frequent malignant tumor was the mixed carcinoma (27.4%). Size dog, breed, housing, and overweight are predictors of malignant mammary tumors in dogs. The highest risk of developing malignant mammary tumors is associated with large female dogs, Yorkshire or Poodle breeds, living outside the home, or being overweight. [ABSTRACT FROM AUTHOR]

MMTV causes mammary tumors in mice, and it is associated with invasive and aggressive forms of breast cancer in humans. However, the underlying mechanisms are yet unknown. We aimed to determine the MMTV-like virus (MMTV-LV) association with histological types of breast cancer, nodal involvement, and metastasis. First, 105 breast cancer biopsies and 15 disease-free biopsies were collected. Details of clinicopathological characteristics were retrieved from patients' records. The status of MMTV-LV was already known for these biopsy samples. Associations of MMTV-LV prevalence with LNM status and metastatic history were determined. Next, quantitative PCR (qPCR) was used to quantify env gene mRNA in biopsies positive for MMTV-LV. Expression of the env gene was compared against different histopathological types of mammary tumor, LNM status, and metastasis by performing Ordinary One Way ANOVA followed by Tukey's multiple comparisons test. MMTV-LV prevalence was found to have no significant association with LNM or metastatic history. As compared to normal control, expression of the env gene was significantly higher (>2.8 folds) in invasive samples (P-value: < 0.01). Expression was also higher (3.28 and 2.89 folds) in patient samples with LNM (P-value: 0.0006) or metastatic history (P-value: < 0.0001), respectively. We conclude that MMTV-LV prevalence is not associated with LNM status or breast cancer metastasis; samples with invasive phenotypes, nodal involvement, and metastasis exhibit significantly higher expression of the MMTV-like env gene. [ABSTRACT FROM AUTHOR]

Aim: This study, the cut-off point of biochemical parameters in bitches with mammary tumors was determined and its usability as a biomarker for diagnosis was investigated. Materials and Methods: Bitches that presented to our clinic for routine ovariohysterectomy (control group, n = 14) and suffer from mammary tumors (CMT group, n = 14) were included in the study. Biochemical parameters including albümin (ALB), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gammaglutamyltransferase (GGT), total protein (TP), direct bilirubin (DB), total bilirubin (TP), creatine, creatine kinase-N-acetyl-cysteine (CK-NAC), urea, phosphorus (P), magnesium (Mg), and calcium (Ca) concentrations were measured with an automated chemistry analyzer. Results: The ALP (p: 0.048), ALT (p < 0.000), AST (p: 0.043), GGT (p: 0.013), TP (p: 0.010), DB (p: 0.043), TP (p: 0.018), creatine (p < 0.000), P (p < 0.000), and CK-NAC (p < 0.000) concentrations were found to be higher in the CMT group than in the control group. Although Mg (p < 0.000) concentration was higher in the control group, there was no statistical significance for ALB, urea, and Ca concentrations among the groups (p > 0.05). It was seen that ALP, ALT, AST, GGT, TP, and creatine concentrations could be used to diagnose mammary tumors, with area under the curve (AUC) values of 0.656, 0.831, 0.610, 0.792, 0.942, and 0.974, respectively. Conclusion: It was concluded that ALP, ALT, AST, GGT, TP, and creatine could be helpful diagnostic tools for canine mammary tumors.

Simple Summary: In this study we addressed the analysis of human breast cancer and canine and feline mammary tumors with regard to the expression, at either gene or protein level, of some molecules that are related to the capacity of an epithelial cell to become mesenchymal (epithelial-to-mesenchymal transition), acquiring higher ability to metastasize. In our samples, some typical markers of this transition were not higher at mRNA levels in tumors than in healthy tissues, indicating that some other markers should be investigated. Instead, at protein levels, some molecules such as vimentin and E-cadherin were indeed associated with higher aggressiveness, being potential useful markers. As already described in the literature, we also demonstrated that feline mammary tumors are close to an aggressive subtype of human breast cancer called triple negative, whereas canine mammary tumors are more similar to the less aggressive subtype of human breast cancer that expresses hormonal receptors. Epithelial-to-mesenchymal transition (EMT) is a process by which epithelial cells acquire mesenchymal properties. EMT has been closely associated with cancer cell aggressiveness. The aim of this study was to evaluate the mRNA and protein expression of EMT-associated markers in mammary tumors of humans (HBC), dogs (CMT), and cats (FMT). Real-time qPCR for SNAIL, TWIST, and ZEB, and immunohistochemistry for E-cadherin, vimentin, CD44, estrogen receptor (ER), progesterone receptor (PR), ERBB2, Ki-67, cytokeratin (CK) 8/18, CK5/6, and CK14 were performed. Overall, SNAIL, TWIST, and ZEB mRNA was lower in tumors than in healthy tissues. Vimentin was higher in triple-negative HBC (TNBC) and FMTs than in ER+ HBC and CMTs (p < 0.001). Membranous E-cadherin was higher in ER+ than in TNBCs (p < 0.001), whereas cytoplasmic E-cadherin was higher in TNBCs when compared with ER+ HBC (p < 0.001). A negative correlation between membranous and cytoplasmic E-cadherin was found in all three species. Ki-67 was higher in FMTs than in CMTs (p < 0.001), whereas CD44 was higher in CMTs than in FMTs (p < 0.001). These results confirmed a potential role of some markers as indicators of EMT, and suggested similarities between ER+ HBC and CMTs, and between TNBC and FMTs. [ABSTRACT FROM AUTHOR]

Copyright of Eurasian Journal of Veterinary Sciences is the property of Eurasian Journal of Veterinary Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The heterogeneous nature of human breast cancer (HBC) can still lead to therapy inefficacy and high lethality, and new therapeutics as well as new spontaneous animal models are needed to benefit translational HBC research. Dogs are primarily investigated since they spontaneously develop tumors that share many features with human cancers. In recent years, different natural phytochemicals including berberine, a plant alkaloid, have been reported to have antiproliferative activity in vitro in human cancers and rodent animal models. In this study, we report the antiproliferative activity and mechanism of action of berberine, its active metabolite berberrubine, and eight analogs, on a canine mammary carcinoma cell line and in transgenic zebrafish models. We demonstrate both in vitro and in vivo the significant effects of specific analogs on cell viability via the induction of apoptosis, also identifying their role in inhibiting the Wnt/β-catenin pathway and activating the Hippo signals with a downstream reduction in CTGF expression. In particular, the berberine analogs NAX035 and NAX057 show the highest therapeutic efficacy, deserving further analyses to elucidate their mechanism of action more in detail, and in vivo studies on spontaneous neoplastic diseases are needed, aiming at improving veterinary treatments of cancer as well as translational cancer research.

Mammary tumors are a common pathology among female cats and dogs, and less common in males of these species. Intact and older females are more susceptible to this condition. A mammary tumor is typically suspected when a lump is discovered during a physical examination of the abdominal area, along the mammary chains. Although surgical treatment is the therapy of choice in most cases, sometimes chemotherapy may be also required. Establishing a correct diagnosis is the foundation for individual optimisation of treatment, as well as the possibility to predict the course of the pathology and prognosis. Diagnosis is based on obtaining an accurate history, clinical, paraclinical and imaging examination of the patient. All this will allow a tumor staging, which will guide therapy and prognosis. This article presents a diagnostic and therapeutic algorithm developed after centralizing the latest data published in the literature, providing a guide in the management of mammary tumors in canids and felines. [ABSTRACT FROM AUTHOR]

Canine mammary tumors represent one of the leading malignant pathologies in female dogs, displaying the importance of efficient therapeutic findings, besides the golden-standard surgery, able to limit the development of the disease. Studies in human cancers demonstrated that Doxorubicin presents a good effect in different biological processes like apoptosis, autophagy, the cell cycle, cell invasion, and the epithelial-to-mesenchymal transition. This study followed the effects of Doxorubicin on two canine mammary cancer cell lines P114 and CMT-U27. Doxorubicin treatment in both cell lines shows an inhibitory effect in cell proliferation and an alteration in expression of the EMT-related genes. The obtained results provide valuable information for revealing the link between Doxorubicin, phenotypic changes, and proliferation dynamics in canine mammary tumor models.

Mammary cancer is one of the main causes of death in female dogs worldwide, considering that many risk factors are involved in its development. This study aimed to elucidate the relationship between epidemiological and clinical risk factors with the histopathological diagnosis of malignant mammary tumors in dogs treated at the Veterinary Hospital of the Federal University of Uberlândia, which has one of the first veterinary oncology services in Brazil. A retrospective matched case-control study was conducted to identify risk factors for the development of malignant mammary tumors in dogs. The variables analyzed were size dog, breed, housing, type of diet, and body score. Potential risk factors were selected by univariate analysis (p < 0.25) before multivariate forward binary logistic regression. The most frequent benign tumor was the benign mixed tumor (35.2%), and the most frequent malignant tumor was the mixed carcinoma (27.4%). Size dog, breed, housing, and overweight are predictors of malignant mammary tumors in dogs. The highest risk of developing malignant mammary tumors is associated with large female dogs, Yorkshire or Poodle breeds, living outside the home, or being overweight.

Carcinoma in situ of the breast is a well-known entity in humans. In veterinary medicine, particularly in canine and feline mammary literature, there is no agreement whether the term in situ should be used to indicate a specific carcinoma histotype or the noninvasive status of a carcinoma of any histotype. Moreover, in the most recent histologic classification of mammary tumors published by the Davis-Thompson Foundation, it is suggested to abandon the term carcinoma in situ given the lack of standardized criteria defining this entity, replacing it with epitheliosis or ductal/lobular hyperplasia with severe atypia. This publication presents a critical review of the term in situ in human and veterinary medicine considering the evolution of the term over the years and its heterogeneous use by different authors, including variations in immunohistochemical markers for classification. This review aims to point out the lack of uniformity in the nomenclature and classification issues in veterinary medicine regarding the use of the term in situ, laying the ground for a process of standardization in future publications. [ABSTRACT FROM AUTHOR]

ABSTRACT: Obesity may be associated with the onset of mammary tumors in women. In companion animals, these data are still scarce, mainly associating the clinic of the patient with laboratory and histological findings. This study aimed to evaluate the clinical and laboratory aspects of female dogs with mammary neoplasia, investigating whether they are related to obesity. Four groups of spayed or non-spayed female dogs were studied, divided into (G1) female dogs without mammary tumor and normal body condition score (BCS), (G2) female dogs without mammary tumor and obese, (G3) female dogs with mammary tumor and normal BCS and (G4) female dogs with mammary tumor and obese. BCS, canine body mass index (CBMI), blood count, glucose, total cholesterol, triglycerides and cytopathological and histopathological examinations were evaluated. Mixed-breed animals were the most prevalent in this study (67.5%). There was a limitation regarding the use of CBMI related to body proportionality. Considering the 28 tumors evaluated, carcinoma in mixed tumor was the most prevalent malignant histological type, while adenomyoepithelioma was the most prevalent benign histological type. It was possible to conclude that obesity was not related to clinical and laboratory changes in the female dogs affected with mammary neoplasias. In addition, no relationship was noted between BCS and CBMI with the histological type or malignancy of mammary neoplasia, since the malignant histological types were the most prevalent tumors in both the ideal score group and the overweight to obese group.

ABSTRACT: Mammary tumors in female dogs are the most frequent and corresponds to half of the canine tumors. The objectives of this study were to determine the risk factors associated with the occurrence of mammary tumors in female dogs and to evaluate the macroscopic characteristics of these neoformations, using 386 dogs from the “Outubro Rosa Pets” events done within the cities of Uberlândia and Patos de Minas, Minas Gerais State, Brazil, in 2015 (n=194), 2016 (n=105) and 2017 (n=87). For the determination of risk factors, the binary logistic regression test (P5cm). At least 15.4% (14/91) of the dogs had one of the regional lymph nodes increased. In conclusion, the occurrence of mammary tumors in the evaluated population was 23.6% and that age, overweight and non-realization of ovariohysterectomy are risk factors associated with the development of mammary tumors.

The aim of this article is to present different view on the gonadectomy of female dogs, for decades considered as gold standard in prevention of gynaecological disorders and also mammary gland tumors. Recently however, this routine practice is increasingly perceived as controversial procedure. Ovariectomy will certainly remain routine surgery in small animals, but the optimal bitch age for spaying may be dependent upon several factors, like breed, body weight, expected lifespan and other breed-related conditions. The risk of developing mammary tumors increases, starting with the female age of 6 years. Ovariectomy or ovariohysterectomy carried out prophylactically in bitches is usually recommended before the first cycle. Investigations however, are necessary to estimate the real benefits and disadvantages of ovariectomy, especially if performed in very young females. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Mammary tumors, for which mastectomy is the main treatment, are the most common neoplasms in bitches. Mastectomy is painful and, in order to reduce the pain stimulus in the transoperative period, tumescent local anesthesia is associated with general inhalation anesthesia. However, despite the numerous benefits of tumescence, intraoperative hypothermia is the most common complication. In Medicine, especially in plastic and dermatological surgery, it is common to use a heated tumescence solution to prevent intraoperative hypothermia; however, in Veterinary medicine, no previous study has examined the advantages and disadvantages of using heated tumescence solution. Thus, this study aimed to investigate the transanesthetic cardiorespiratory effects of heated tumescence solution in bitches submitted to radical unilateral mastectomy. Materials, Methods & Results: Eight animals were treated with 0.1% lidocaine solution, warmed to 37-42°C, using a Klein’s cannula for administration. Chlorpromazine (0.3 mg/kg) and meperidine (3 mg/kg) were used as pre-anesthetic medication intramuscularly, and induction was performed with intravenous propofol and maintenance with isoflurane. The data collection times were as follows: 15 min after starting isoflurane administration (M1), 5 min after tumescence (M2), after beginning of surgical incision (M3), during breast pullout (M4), after clamping of the superficial caudal epigastric vein, and artery (M5), after the beginning of the approximation of the subcutaneous tissue (M6), after the beginning of the intradermal suture (M7), and at the end of the surgical procedure (Mfinal). The heart (HR) and respiratory (ƒ) rates, mean arterial pressure (MAP), end-tidal CO2 concentration (EtCO2 ), expired isoflurane concentration (EtISO), and rectal temperature (RT) were measured. The HR, ƒ, and EtCO2 levels did not differ statistically. The mean EtISO presented in M2 (1.16 ± 0.41) was significantly lower than that in M3 (1.39 ± 0.40) and M4 (1.49 ± 0.49). Discussion: In the HR analysis, it was found that during all evaluation moments, the means remained within the reference range for the species. Moreover, the values during the breast pullout (M4) did not exceed 20% of those presented minutes before the beginning of the surgery (M2), which was indicative of analgesic rescue, suggesting that the animals did not experience pain. Hypoventilation resulted in an increase in EtCO2 values. Thus, it can be said that in this study, there was no respiratory depression during the transoperative period, as the values of the variables ƒ and EtCO2 were within the reference for the species. With regard to the EtISO variable, there was no reduction in the MAC of isoflurane with the use of heated tumescence solution, as reported by some authors (EtISO 0.8%). However, the EtISO values presented here are close to those found in the literature during breast pullout (EtISO between 1.3% and 1.52%), with the use of refrigerated tumescence solution. In addition, the values shown in M4 are within the equivalent of 1 MAC (1.41%) of isoflurane, proving that heated tumescent local anesthesia is a safe technique and an excellent adjunct to inhalation anesthesia, as it provides intraoperative analgesia. Therefore, heated tumescence solution is safe and an excellent adjuvant in general inhalational anesthesia for radical unilateral mastectomy as it did not increase inhaled anesthetic consumption during surgery. [ABSTRACT FROM AUTHOR]

Copyright of Al-Anbar Journal of Veterinary Sciences is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract Background PIK3CA mutations are frequent in human breast cancer. Pik3caH1047R mutant expression in mouse mammary gland promotes tumorigenesis. TP53 mutations co-occur with PIK3CA mutations in human breast cancers. We previously generated a conditionally activatable Pik3caH1047R;MMTV-Cre mouse model and found a few malignant sarcomatoid (spindle cell) carcinomas that had acquired spontaneous dominant-negative Trp53 mutations. Methods A Pik3caH1047R;Trp53R270H;MMTV-Cre double mutant mouse breast cancer model was generated. Tumors were characterized by histology, marker analysis, transcriptional profiling, single-cell RNA-seq, and bioinformatics. Cell lines were developed from mutant tumors and used to identify and confirm genes involved in metastasis. Results We found Pik3caH1047R and Trp53R270H cooperate in driving oncogenesis in mammary glands leading to a shorter latency than either alone. Double mutant mice develop multiple histologically distinct mammary tumors, including adenocarcinoma and sarcomatoid (spindle cell) carcinoma. We found some tumors to be invasive and a few metastasized to the lung and/or the lymph node. Single-cell RNA-seq analysis of the tumors identified epithelial, stromal, myeloid, and T cell groups. Expression analysis of the metastatic tumors identified S100a4 as a top candidate gene associated with metastasis. Metastatic tumors contained a much higher percentage of epithelial–mesenchymal transition (EMT)-signature positive and S100a4-expressing cells. CRISPR/CAS9-mediated knockout of S100a4 in a metastatic tumor-derived cell line disrupted its metastatic potential indicating a role for S100a4 in metastasis. Conclusions Pik3caH1047R;Trp53R270H;MMTV-Cre mouse provides a preclinical model to mimic a subtype of human breast cancers that carry both PIK3CA and TP53 mutations. It also allows for understanding the cooperation between the two mutant genes in tumorigenesis. Our model also provides a system to study metastasis and develop therapeutic strategies for PIK3CA/TP53 double-positive cancers. S100a4 found involved in metastasis in this model can be a potential diagnostic and therapeutic target.

Background: Breast cancer is the most common cancer in women worldwide and is the leading cause of death amongst women with cancer. One novel therapy used for breast cancer treatment constitutes non-contact electric fields and is called electro-capacitive cancer therapy (ECCT) with intermediate frequency and low intensity. The objective of this study was to examine the effect of ECCT on mammary tumors growth in rats and observing the immune responses that play a role in fighting the tumor. Methods: Female SD rats were used and divided into four groups, namely control (NINT), placebo (NIT), non- therapy (INT), and therapy (IT) groups with 6 biological replicates in each group. Rats in INT and IT groups were treated with 7,12-dimethylbenz[a]anthracene for mammary tumor induction. Only rats in NIT and IT groups were exposed to ECCT individually for 10 hours per day for 21 days. The size of all tumors was measured with a digital caliper. The distributions of PCNA, ErbB2, caspase-3, CD68, CD4, and CD8-positive cells were observed with immunohistochemistry and scoring with ImageJ. Results: The growth rate of mammary tumors in IT group was significantly lower (p Conclusions: Non-contact electric fields exposure showed inhibition on mammary tumor growth in rats while inducing CD8+ T cells, leading to tumor cell death and potentially helping wounds heal.

Background: Breast cancer is the most common cancer in women worldwide and is the leading cause of death in women with cancer. One novel therapy used for breast cancer treatment is non-contact electric fields called electro-capacitive cancer therapy (ECCT) with intermediate frequency (100 kHz) and low intensity (18 Vpp). The objective of this study was to examine the effect of ECCT on mammary tumors growth in rats and observing the immune responses that play a role in fighting the tumor. Methods: Female SD rats were used and divided into four groups, namely control (NINT), placebo (NIT), non- therapy (INT), and therapy (IT) groups with 6 biological replicates in each group. Rats in INT and IT groups were treated with 7,12-dimethylbenz[a]anthracene for mammary tumor induction. Only rats in NIT and IT groups were exposed to ECCT individually for 10 hours per day for 21 days. The size of all tumors was measured with a digital caliper. The distributions of PCNA, ErbB2, caspase-3, CD68, CD4 and CD8-positive cells were observed with immunohistochemistry and scoring with ImageJ. Results: The growth rate of mammary tumors in IT group was significantly lower (p Conclusions: Non-contact electric fields exposure showed inhibition on mammary tumor growth in rats while inducing CD8+ T cells that lead to tumor cells death and potentially helps wound healing.

ABSTRACT: The result of the reaction of free radicals with biomolecules is the formation of substances with the potential of inducing oxidative damage, a condition known as oxidative stress. There are voluminous literature data reporting the association, both as a cause and as a consequence, between different diseases and oxidative stress. In this study, 144 female dogs with mammary neoplasia were analyzed. The animals were submitted to clinical evaluation for disease staging, hematological evaluation, serum biochemistry (renal and hepatic function tests), and dosage of the oxidative damage biomarker, malondialdehyde (MDA), at the time of its approach and 30 days after treatment. A control group of 100 healthy animals was also submitted to determination of serum MDA levels. The mean age of the animals affected by mammary neoplasms was 9.88±2.95 (4 to 14) years, while in healthy animals it was 2.31±1.90 years (1 to 6). Of the 144 animals, 113 (78.9%) had malignant neoplasms, and 15, 21, 46, 17 and 14 animals were in clinical stage I, II, III, IV and V respectively and the carcinoma in a mixed tumor was the most frequent histological pattern in this group (26%). Thirty-one animals were diagnosed with benign neoplasms and mammary adenoma was the most frequent histological pattern in 15 animals (51.61%). Hematological changes in the preoperative period were observed in 44 (38.9%) and 12 (38.7%) animals with malignant and benign neoplasias, respectively, and there was a positive correlation between anemia and higher levels of MDA (P=0.0008) for animals with malignant tumors. Regarding serum biochemical parameters, the most frequent alterations in animals with malignant neoplasms were elevated ALT levels in 12 animals (10.6%), creatinine in 10 animals (8.84%) and urea in eight animals (7.07%). Females with benign neoplasms presented less occurrence of changes in these parameters. In the group of healthy animals (control), the mean serum MDA values were 12.08±4.18, whereas in the pre-treatment group, mean MDA was 24.80±5.74 for bitches with benign neoplasms and 32.27±10.24 for bitches with malignant tumors. A significant increase (P

1,2‐dichloroethane (DCE or EDC) is a chlorinated hydrocarbon used as a chemical intermediate, including in the synthesis of polyvinyl chloride. Although DCE has induced tumors in both rats and mice, the overall weight‐of‐evidence suggests a lack of in vivo mutagenicity. The present study was conducted to explore a potential mode of action further for tumor formation in rat mammary tissue. Fischer 344 rats were exposed to target concentrations of 0 or 200 ppm of DCE vapors (6 hours/day, 7 days/week) for at least 28 days; 200 ppm represents a concentration of ~20% higher than that reported to induce mammary tumors. Endpoints examined included DNA damage (via Comet assay), glutathione (reduced, oxidized and conjugated), tissue DNA adducts, cell proliferation and serum prolactin levels. Exposure to DCE did not alter serum prolactin levels with consistent estrous stage, did not cause cell proliferation in mammary epithelial cells, nor result in histopathological alterations in the mammary gland. DNA adducts were identified, including the N7‐guanylethyl glutathione adduct, with higher adduct levels measured in liver (nontumorigenic target) compared with mammary tissue isolated from the same rats; no known mutagenic adducts were identified. DCE did not increase the Comet assay response in mammary epithelial cells, whereas DNA damage in the positive control (N‐nitroso‐N‐methylurea) was significantly increased. Although the result of this study did not identify a specific mode of action for DCE‐induced mammary tumors in rats, the lack of any exposure‐related genotoxic responses further contributes to the weight‐of‐evidence suggesting that DCE is a nongenotoxic carcinogen. 1,2‐dichloroethane (DCE or EDC) is a chlorinated hydrocarbon used as a chemical intermediate, including in the synthesis of polyvinyl chloride. The present study was conducted to explore further the potential mode of action for tumor formation in rat mammary tissue. Although the results of this study do not identify a specific mode of action for DCE‐induced mammary tumors in rats, the lack of any exposure‐related genotoxic responses further contributes to the weight‐of‐evidence suggesting that DCE is a nongenotoxic carcinogen. [ABSTRACT FROM AUTHOR]

Breast cancer is the most frequent neoplasia in developed countries and the leading cause of death in women worldwide. Epithelial-to-mesenchymal transition (EMT) is a cellular process through which epithelial cells decrease or lose their epithelial characteristics and gain mesenchymal properties. EMT mediates tumor progression, because tumor cells acquire the capacity to execute the multiple steps of invasion and metastasis. Benzo[a]pyrene (B[a]P) is an environmental organic pollutant generated during the burning of fossil fuels, wood, and other organic materials. B[a]P exposition increases the incidence of breast cancer, and induces migration and/or invasion in MDA-MB-231 and MCF-7 breast cancer cells. However, the role of B[a]P in the induction of an EMT process and metastasis of mammary carcinoma cells has not been studied in detail. In this study, we demonstrate that B[a]P induces an EMT process in MCF10A mammary non-tumorigenic epithelial cells. In addition, B[a]P promotes the formation of larger tumors in Balb/cJ mice inoculated with 4T1 cells than in untreated mice and treated with dimethyl sulfoxide (DMSO). B[a]P also increases the number of mice with metastasis to brain and the total number of brain metastatic nodules in Balb/cJ mice inoculated with 4T1 cells compared with untreated mice and treated with DMSO. In conclusion, B[a]P induces an EMT process in MCF10A cells and the growth of mammary tumors and metastasis to brain in Balb/cJ mice inoculated with 4T1 cells., (© 2024 The Authors. Molecular Carcinogenesis published by Wiley Periodicals LLC.)

The present study estimated the level of some macro and micro minerals in female dogs with mammary gland tumors fed with different habitual diets. Female dogs bearing malignant mammary tumors were divided into two groups; group-I (n=12) fed only with balanced commercial diet and group-II (n=12) fed only with homemade diet. Ten physiologically healthy female dogs were taken as control. Trace elements viz. iron, copper, zinc and manganese were estimated in plasma and mammary tumor tissues. Calcium and phosphorus were exclusively estimated in plasma. Plasma concentrations of iron, copper and calcium were found to be significantly (p

Reduction of expression of E-cadherin in canine mammary tumors has been correlated with invasive behavior, lymph node metastases and shorter survival period after tumor removal. The aim of this study was to examine the expression of E-cadherin in canine benign and malignat mammary tumors with different grade of malignancy. 38 canine mammary tumors were studied. In general, the percentage of positive stained cells was bigger in benign neoplasia and carcinomas with grade of malignancy The expression of E-Cadherin decreased with growth of the grade of malignancy. Among the groups of tumors of different grade of malignancy, statistical significant loss of expression of E-Cadherin was detected between benign lesions and carcinomas grade of malignancy II (p< 0,01), and between benign lesions and carcinomas III grade of malignancy (p< 0,01). [ABSTRACT FROM AUTHOR]

Breast cancer is the most prevalent tumor type among women and female dogs. Tumor malignancy is characterized by the epithelial-to-mesenchymal transition (EMT) which leads to the metastasis formation. The inhibition of angiotensin II type I receptor (AGTR1) by an antagonist such as losartan can suppress angiogenesis, consequently contributing to the metastasis control. The aim of this study was to analyze the capacity of losartan and AGTR-1 gene edition to modulate the EMT process in triple negative/metastatic mammary tumor cells, compared to existing treatment protocols such as carboplatin. The cell lines CF41.Mg and MDA-MB-468, were cultured and treated with carboplatin, losartan, or submitted to AGTR-1 gene edition by CRISPR/Cas9. EMT markers and PARP-1 protein and gene expression were evaluated by immunofluorescence or immunocytochemistry and qRT-PCR, respectively. Cell migration capacity was also evaluated. For CF41.Mg and MDA-MB-468 cell lines, there was an increase in E-cadherin and a decrease in N-cadherin and PARP-1 protein and gene expression after treatment with carboplatin, losartan, both in combination and after AGTR-1 gene edition. There was a decrease in VEGF and PARP-1 protein and gene expression after AGTR-1 gene edition. Moreover, in both lines, reduction in invasion rate was observed after all treatments. Our data suggest that losartan and the gene edition of AGTR-1 by CRISPR/Cas9 were able to block the DNA repair and control the EMT process, such as carboplatin. The results in the canine species are unprecedented, as there are no data in the literature that demonstrate the action of losartan in this tumor type.

Elastography is a sonographic technique that provides a noninvasive evaluation of the stiffness of a lesion. The objective of this work was to evaluate the accuracy of strain elastography, the most accessible modality in clinical practice, to discriminate between different histological types of malignant mammary neoplasms in the canine species, which can provide complementary information in real time to the diagnosis and thus help in the choice of surgical technique. A total of 34 females with 56 mammary carcinomas were selected and classified into three histological groups according to their aggressiveness. The histological and elastographic characteristics of these malignant tumors were analyzed and compared to evaluate the diagnostic accuracy of strain elastography. Visual score presented a sensitivity of 88.0%, specificity of 58.1%, and accuracy of 71.43% in distinguishing the most aggressive group of carcinomas. The strain ratio had a sensitivity of 84.0%, specificity of 61.1%, and accuracy of 69.64%. On the other hand, intratumoral strain ratio obtained a sensitivity of 71.40% and specificity of 61.90% when intratumoral fibrosis was taken as reference, with an accuracy of 66.07%. Similarly, peritumoral strain ratio was also positively related to fibrosis in the periphery of lesions (p ≤ 0.001), with a sensitivity of 93.80%, specificity of 77.50% and an accuracy of 92.87%. In conclusion, accuracy of this elastographic modality can be a useful method to differentiate more aggressive histological types. Therefore, it represents an additional diagnostic technique useful in the daily clinic thanks to the short time required for the examination, which allows real-time visualization and immediate interpretation of the results.

Canine malignant mammary tumors (MMTs) are prevalent malignancy in intact female dogs with a high incidence of metastasis and recurrence. A current lack of easily accessible tumor biomarkers hinders a timely assessment of the disease outcome. We previously identified anterior gradient protein 2 (AGR2) with higher protein abundance in canine MMT tissues compared with normal counterparts. AGR2 is an endoplasmic reticulum-resident protein disulfide isomerase involved in the regulation of protein processing and also exists extracellularly via secretion to exert pro-oncogenic functions. In the present study, we validated overexpression of AGR2 in canine MMT tissues from 45 dogs using immunohistochemistry and immunoblotting, and assessed serum AGR2 levels in 81 dogs with MMTs and 21 benign cases using a competitive enzyme-linked immunosorbent assay (ELISA). Our data revealed that serum eAGR2 levels are significantly correlated with MMT progression (p = 0.0007) and remote tumor metastasis (p = 0.002). Moreover, elevated levels of serum eAGR2 are associated with an unfavorable overall survival of MMT dogs in later stage (p = 0.0158). Area under the time-dependent ROC curve (AUC) of serum eAGR2 level as a prognostic indicator was 0.839. Collectively, this study uncovered that serum eAGR2 level is significantly associated with an adverse outcome of MMT dogs and holds a predictive potential in MMT prognosis.

Objective- Breast cancer is one of the most serious problems in oncology. Alkaloids pyrrolizidine are widely found in nature and exhibited versatile biological activities. The aim of the present study was to assess anti-tumor activity of methyl 1,3-dioxo-1,1',2',3,5',6',7',7a'-octahydrospiro[indene-2,3'-pyrrolizidine]-2'-carboxylate (6) on 7,12 dimethylbenz(a) anthracene (DMBA)-induced mammary tumors in rat. Design- Experimental study. Animals- Twenty-one female Wistar ats Procedures- 7-Week-old female Wistar rats (180 g body weight) were randomized into three groups of seven animals each. DMBA-induced mammary tumors was achived using DMBA dissolved in 1ml of vehicle (0.5 ml of DMSO plus 0.5 ml of saline) and injected by subcutaneous injection beneath the mammary gland on either side. Tumor yield and size were stabilized after 90 days with the initiation of DMBA. Healthy intact animals (NC) were considered as a normal control feeding on pellets and tap water. In group DMBA, the tumor was induced by DMBA. In group DMBA/7d the animals with tumor received 100 µL 6 dissolved in DMSO (0.25 µM) solution intraperitoneally for one week. Synthesis of 6 was accomplished and the structure of new product was assigned by their 1H/13C NMR, IR, CH-COSY and mass spectral data as well as the elemental analysis. Results- The overall tumor analysis showed that 6 treatment significantly inhibited the breast tumor incidence, tumor multiplicity, and tumor size in the DMBA-initiated rat model. 6 treatment significantly inhibited the total volume of tumors per rat. Conclusion and Clinical Relevance-The findings of the present study showed that the 6 treatment was significantly effective in reduction of tumors versus the cancer controls.

Aim: The main objective of the study was to describe the use of skin fold advancement flaps (SFAFs) and other reconstructive techniques for closure of large skin defects following mammary tumor excision in dogs. Materials and Methods: Twelve dogs underwent reconstruction of large ventral skin defects following mammary tumor excision with wide margins. Skin fold flaps (flank fold flap and elbow fold flap) were elevated from the flank and elbow region, respectively, and transposed and sutured onto the large ventral skin defect following mastectomy in all the dogs. In addition to the skin fold flaps, other reconstructive techniques such as undermining, walking sutures, and tension-relieving suture techniques were followed during surgery in the closure of large skin defects without skin tension and compromising limb mobility. The skin flap viability was assessed subjectively by gross observation of the flap such as color, temperature, capillary perfusion, and cosmetic appearance, and scoring (1-4) was done. Tissue samples were collected from a surgical site on days 3, 6, and 12 post-operatively for histopathological evaluation and healing status of the skin flap. Results: All the surgical wounds healed primarily, without any major complications and the skin flap remained healthy throughout the healing process post-operatively. Distal flap necrosis was noticed in one case and necrosis of skin flap between two suture lines was noticed in another case in which the necrotized distal portion healed by secondary intention after 7 days. The mean survival of subdermal plexus flap in the above cases was 98% which was a subjective evaluation based on surface area of the skin defect measured by Image 'J software and the flap dimensions. The average healing of skin flap in days was 14.91±0.86. Conclusion: The SFAFs along with other reconstructive techniques help in the reconstruction of large ventral skin defects following mastectomy in dogs without much compromising limb mobility.

Background: PIK3CA mutations are frequent in human breast cancer. Pik3caH1047R mutant expression in mouse mammary gland promotes tumorigenesis. TP53 mutations co-occur with PIK3CA mutations in human breast cancers. We previously generated a conditionally activatable Pik3caH1047R;MMTV-Cre mouse model and found a few malignant sarcomatoid (spindle cell) carcinomas that had acquired spontaneous dominant-negative Trp53 mutations.Methods: A Pik3caH1047R;Trp53R270H;MMTV-Cre double mutant mouse breast cancer model was generated. Tumors were characterized by histology, marker analysis, transcriptional profiling, single-cell RNA-seq, and bioinformatics. Cell lines were developed from mutant tumors and used to identify and confirm genes involved in metastasis.Results: We found Pik3caH1047R and Trp53R270H cooperate in driving oncogenesis in mammary glands leading to a shorter latency than either alone. Double mutant mice develop multiple histologically distinct mammary tumors, including adenocarcinoma and sarcomatoid (spindle cell) carcinoma. We found some tumors to be invasive and a few metastasized to the lung and/or the lymph node. Single-cell RNA-seq analysis of the tumors identified epithelial, stromal, myeloid, and T cell groups. Expression analysis of the metastatic tumors identified S100a4 as a top candidate gene associated with metastasis. Metastatic tumors contained a much higher percentage of epithelial-mesenchymal transition (EMT)-signature positive and S100a4-expressing cells. CRISPR/CAS9-mediated knockout of S100a4 in a metastatic tumor-derived cell line disrupted its metastatic potential indicating a role for S100a4 in metastasis.Conclusions: Pik3caH1047R;Trp53R270H;MMTV-Cre mouse provides a preclinical model to mimic a subtype of human breast cancers that carry both PIK3CA and TP53 mutations. It also allows for understanding the cooperation between the two mutant genes in tumorigenesis. Our model also provides a system to study metastasis and develop therapeutic strategies for PIK3CA/TP53 double-positive cancers. S100a4 found involved in metastasis in this model can be a potential diagnostic and therapeutic target. [ABSTRACT FROM AUTHOR]

Mammary neoplasms are a heterogeneous form of disease, and in order to determine its course and biological features with more accuracy, investigations based on tumor phenotypes are required. The aim of the present study was to propose and validate a phenotypic classification for canine mammary tumors and to assess any association between clinicopathological characteristics, survival and prognosis. For the immunohistochemistry analysis, the primary antibodies against estrogen receptor-α, progesterone receptor, human epidermal growth factor receptor 2 (HER-2)/neu and E-cadherin were used. A total of 110 canine mammary tumors were investigated; 42 tumors were classified as luminal A, 41 as luminal B, 17 as triple-negative and 10 as HER-2-positive. The luminal A and B phenotypes were associated with improved prognosis, whereas HER-2positive and triple-negative tumors were more aggressive, and exhibited a significant association with the occurrence of metastasis, a worse Tumor-Node-Metastasis classification and shorter survival time (P<0.05). In addition, there were different levels of E-cadherin expression intensity observed among the four tumor profiles investigated. Luminal A and B phenotypes presented an upregulation of E-cadherin compared with the HER-2 positive and triple-negative phenotypes (P<0.05). From the results of the present study, the proposed immunohistochemical panel and phenotypic classification techniques could be useful diagnostic tools with a good technical applicability in veterinary oncology. The analysis of E-cadherin expression in the panel of tumor markers allowed a more accurate classification for determining the biological features of the mammary tumor. [ABSTRACT FROM AUTHOR]

Parameters of 37 canine mammary malignant tumors and 13 dysplasia were compared. The age of an animal at the time of treatment in veterinary clinic, the age of the first factors of neoplasm, the number of births during life and the period of the last one, feeding nature, pseudo pregnancy and lactation cases, the number and location of affected mammary gland, neoplasm macro characteristics were considered. The size of the letter was determined with a view to three factors, the occurrence of capsules in neoplasms was ranked in points. The intensity of lactation malfunction was determined under the same method. Canine mammary dysplasia was examined at an earlier age than malignant tumors. This is partly due to the earlier onset than in the case of malignant tumors. At the same time, period between cancer discovery and treatment in the latter case was much shorter. It may state faster growth of tumors in case of malignancy than in mammary dysplasia. A significant difference in neoplasms size - malignant tumor and dysplasia is the proof thereof. Study of the influence of the number of births in the occurrence of different mammary neoplasms showed that dogs with dysplasia gave birth twice as less than those with diagnosed malignant tumors. The period between the last birth and identification of factors of disease for both groups of animals was the same. There was no significant difference also in the frequency of cases with lactation malfunction in dogs with mammary malignant tumors and dysplasia. Groups of animals with mammary malignant tumors and dysplasia differed on such characteristics as frequency of tumors with full, partial or no capsule, and the average number of mammary gland affected and their localization. [ABSTRACT FROM AUTHOR]

Highlights • Conventional radiotherapy and synchrotron microbeam radiotherapy elicit different tumor immune responses. • Conventional but not microbeam radiotherapy induces infiltration of macrophages and neutrophils into mammary tumors. • In contrast, higher numbers of T cells and pro-inflammatory genes concentrate in tumors following microbeam radiation compared to conventional radiation. Abstract Background Synchrotron microbeam radiation therapy (MRT) is a new, evolving form of radiotherapy that has potential for clinical application. Several studies have shown in preclinical models that synchrotron MRT achieves equivalent tumor control to conventional radiotherapy (CRT) but with significantly reduced normal tissue damage. Methods To explore differences between these two modalities, we assessed the immune cell infiltrate into EMT6.5 mammary tumors after CRT and MRT. Results CRT induced marked increases in tumor-associated macrophages and neutrophils while there were no increases in these populations following MRT. In contrast, there were higher numbers of T cells in the MRT treated tumors. There were also increased levels of CCL2 by immunohistochemistry in tumors subjected to CRT, but not to MRT. Conversely, we found that MRT induced higher levels of pro-inflammatory genes in tumors than CRT. Conclusion Our data are the first to demonstrate substantial differences in macrophage, neutrophil and T cell numbers in tumors following MRT versus CRT, providing support for the concept that MRT evokes a different immunomodulatory response in tumors compared to CRT. [ABSTRACT FROM AUTHOR]