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The present review has analyzed the scientific literature, available in the PubMed and Scopus databases, in order to summarize the current state of diet anthocyanin research in breast cancer (BC) and colorectal cancer (CRC) animal models but also for up-to-date human studies. For CRC, 28 preclinical and 9 clinical studies were selected in line with our search query in science databases. In relation to BC, 14 preclinical and 5 clinical studies were selected. Remarkably, all the preclinical studies, to a greater or lesser degree, suggested a chemoprevention effect of anthocyanin in BC/CRC rodent models. These encouraging results from animal models are not extrapolated to the same degree to human studies where, from the similar theoretical daily doses of anthocyanins in these studies, the opposite results were reported. Nevertheless, it is worth mentioning that the anthocyanin doses in the human studies carried out recently are low if we consider the estimated exposure to anthocyanins issued by the European Food Safety Agency (EFSA) or extremely low if we consider with caution the human equivalent dose based on body surface area from the preclinical dosage regimes used. Therefore, although some clinical data has demonstrated an inverse relation between anthocyanin consumption and BC/CRC, this could, in fact, be more relevant if we increase the daily human anthocyanin dose (as observed in animal model dose-effect studies) while new toxicological data for this flavonoid subtype are brought to light.

This study aimed to evaluate role of nano-sized zinc (Zn) on lactation performance, health status, and mammary permeability of lactating dairy cows. Thirty multiparous dairy cows with similar days in milk (158 ± 43.2) and body weight (694 ± 60.5 kg) were chosen based on parity and milk production and were randomly assigned to 3 treatment groups: basal diet (control, 69.6 mg/kg of Zn adequate in Zn requirement), basal diet additional Zn-methionine (Zn-Met, providing 40 mg/kg of Zn), and basal diet additional nano-sized Zn oxide (nZnO, providing 40 mg/kg of Zn). The study lasted for 10 wk, with the first 2 wk as adaptation. Feed intake, milk yield and the related variables, and plasma variables were determined every other week. Blood hematological profiles were determined in the 8th week of the study. We found that feed intake, milk yield, and milk composition were similar across the 3 groups. The nZnO- and Zn-Met-fed cows had greater milk Zn concentrations in the milk (3.89 mg/L (Zn-Met) and 3.93 mg/L (nZnO)) and plasma (1.25 mg/L (Zn-Met) and 1.29 mg/L (nZnO)) than the control cows (3.79 mg/L in milk and 1.21 mg/L in plasma). The nZnO-fed cows had higher Zn concentrations in plasma but not in milk compared to Zn-Met-fed cows. The Zn appearance in milk was greater in nZnO-fed (area under curve during the first 4 h post-feeding for milk Zn: 16.1 mg/L) and Zn-Met-fed cows (15.7 mg/L) than in control cows (15.0 mg/L). During the first 4 h post-feeding, milk to blood Zn ratio was greater in nZnO-fed animals but lower in Zn-Met-fed cows compared with control cows. Oxidative stress-related variables in plasma, blood hematological profiles, and mammary permeability related variables were not different across treatments. In summary, lactation performance, Zn concentrations in milk and plasma, hematological profiles, mammary permeability were similar in cows fed nZnO and Zn-Met. We therefore suggested that nZnO feeding can improve Zn bioavailability without impairing lactation performance, health status, and mammary gland permeability in dairy cows.

The proteoglycan versican (VCAN) plays a complex role in cancer. The expression of this molecule has been related to invasion and progression in malignant mixed tumors, such as carcinoma in mixed tumors (CMT) of the canine mammary gland. In addition, its interaction with surface cell receptors EGFR, HER-2 and CD44 in malignant epithelial cells may be responsible for proliferation and cellular motility in early stages of cancer. We comparatively evaluated the expression of this proteoglycan and its receptors in in situ and invasive areas of simple carcinomas (SC) and CMT to investigate similarities and differences between these histological types. Immunohistochemistry was performed with anti-VCAN, anti-CD44, anti-EGFR and anti-HER-2 antibodies in 32 cases of SC or CMT. VCAN was highly expressed in stroma adjacent to invasive areas in SC and CMT. CMTs presented comparatively higher expression of VCAN in stroma adjacent to in situ and in invasive areas than in corresponding areas in SCs. In CMT, EGFR and HER-2 expressions were higher in situ compared to invasive areas. In contrast, increased CD44 and EGFR expression was found in invasive areas in SC compared to CMT. These results indicate that versican expression is similarly associated with invasiveness in SC and CMT, however higher levels were seen in CMT suggesting that the presence of myoepithelial proliferation in this tumor type participates in stromal composition and promoting an increase in the expression of versican.

Breast cancer is the leading cause of cancer-related death in women worldwide. Several studies have addressed the association between cancer in humans and agricultural pesticide exposure. Evidence indicates that exposure to organophosphorous pesticides such as parathion and malathion occurs as a result of occupational factors since they are extensively used to control insects. On the other hand, estrogens have been considered beneficial to the organism; however, epidemiological studies have pointed out an increased breast cancer risk in both humans and animals. Experimental female rat mammary gland cancer models were developed after exposure to parathion, malathion, eserine, an acetylcholinesterase inhibitor, and estrogen allowing the analysis of the signs of carcinogenicity as alteration of cell proliferation, receptor expression, genomic instability, and cell metabolism in vivo and in vitro. Thus, pesticides increased proliferative ducts followed by ductal carcinoma; and 17β-estradiol increased proliferative lobules followed by lobular carcinomas. The combination of both pesticides and either eserine or estrogen induced tumors with both types of structures followed by mammary gland tumors and metastasis to the lung and kidneys after 240 days of a 5-day treatment. Studies also showed that these pesticides and eserine decreased three to five times the acetylcholinesterase activity in the serum compared to controls whereas terminal end buds increased in number, being inhibited by atropine. Genomic instability was analyzed in such tissues (mp53, CYP1A2, c-myc, c-fos, ERα, M2R) and pesticides increased protein expression that was stimulated by estrogens but inhibited by atropine. Eserine also transformed the epithelium of the rat mammary gland in the presence of estrogen and increased the number of terminal end buds after treatment inducing mammary carcinomas. Then, enzymatic digestion of such structures gave rise to cells with increased DNA synthesis and induced anchorage independence. Thus, there were changes in the epithelium of the mammary gland influencing breast carcinogenesis. Furthermore, these substances and acetylcholine also showed the signs of carcinogenicity in vitro as cell proliferation, receptor expression (ERα, ErbB2, M2R), genomic instability (c-myc, mp53, ERα, M2R), and cell metabolism. A unique cellular model is also presented here based on the use of MCF-10 F, a non-tumorigenic cell line that represents a valuable clinically translatable experimental approach that identifies mechanistic links for pesticides and estrogen as suspect human carcinogenic agents.

Transgenic breast cancer mouse models are critical tools for preclinical studies of human breast cancer. Genetic editing of the murine mammary gland allows for modeling of abnormal genetic events frequently found in human breast cancers. Genetically engineered mouse models (GEMMs) of breast cancer employ tissue-specific genetic manipulation for tumorigenic induction within the mammary tissue. Under the transcriptional control of mammary-specific promoters, transgenic mouse models can simulate spontaneous mammary tumorigenesis by expressing one or more putative oncogenes, such as MYC, HRAS, and PIK3CA. Alternatively, the Cre-Lox system allows for tissue-specific deletion of tumor suppressors, such as p53, Rb1, and Brca1, or specific knock-in of putative oncogenes. Thus, GEMMs can be designed to implement one or more genetic events to induce mammary tumorigenesis. Features of GEMMs, such as age of transgene expression, breeding quality, tumor latency, histopathological characteristics, and propensity for local and distant metastasis, are variable and strain-dependent. This review aims to summarize currently available transgenic breast cancer mouse models that undergo spontaneous mammary tumorigenesis upon genetic manipulation, their varying characteristics, and their individual genetic manipulations that model aberrant signaling events observed in human breast cancers.

Differentiated mammary epithelial cells are responsible for milk synthesis during lactation, supporting early postnatal life in mammals. These cells are found in the terminal alveoli of a secretory epithelium, which is surrounded by myoepithelial cells and a stroma rich in fatty tissue. The aim of this study was to explore the cell-specific expression of the glucose transporter GLUT8 in mammary gland and evaluate its functionality for glucose transport, in order to confirm its role in lactose synthesis. Our histological results revealed that GLUT8 is expressed in adipocytes and the epithelial and myoepithelial cells in mammary gland, with a predominant intracellular granular pattern. Colocalization studies of endogenous and green fluorescent protein fused GLUT8 revealed their expressions in lysosome and Golgi, respectively, with Pearson's coefficient correlations of 0.82 ± 0.05 and 0.68 ± 0.16. Functional studies of dileucine to dialanine mutant of GLUT8 showed a fructose-sensitive 2-deoxy glucose uptake at a rate of 83.3 pmoles/(min∗106 cells), 7 folds over empty vector, with a 60 ± 4 and 72 ± 6% decline in 2-deoxy glucose in the presence of 20 and 50 mM fructose, respectively. We concluded that functional GLUT8 is expressed in mammary gland, localizing in mammary epithelial and myoepithelial cells, and adipocytes. In lactation, GLUT8 is expressed mainly in luminal epithelial cells, at the compartments of the endomembrane system. It is necessary to explore the physiological/pathological functions of GLUT8 in mammary gland, including its role in lactation.

Fat metabolism is a complex process regulated by a number of factors. Adiponectin receptor 1 (ADIPOR1) gene takes active part in lipid metabolism. Although, there have been some researches indicating that ADIPOR1 could influence the milk fat metabolism through targeting some factors, little is known about the effect of ADIPOR1 on goat milk fat metabolism. To investigate the regulatory role of ADIPOR1 on milk fat metabolism in GMECs, we analysed overexpression in the presence and absence of AdipoRon (50 μM) and examined knockdown using siRNA. Using RT-qPCR, we assessed ADIPOR1 mRNA expressions among different lactation stages in goat mammary gland and the expression of six genes that regulate milk fat metabolism in GMECs.ADIPOR1 mRNA expression level was higher during the various lactation stages, except dry-off period. Knockdown and overexpression results revealed a significant decrease and increase in mRNA expression of ADIPOR1 and genes considered: SREBF1, ACACA, FASN, SCD, ATGL, and HSL, respectively. Treatment of GMECs with AdipoRon 50 μM resulted in a significant (p 0.05) increase in the mRNA expression of all measured genes, except SREBF1.Overall, ADIPOR1 plays a central role in regulating the transcription of several genes involved in milk fat metabolism.

Background: Rabbits, ferrets, and four-toed hedgehogs are popular exotic pets, but comprehensive epidemiological studies in these animals have been rarely conducted. The present study aims to clarify the incidence of neoplastic and non-neoplastic diseases in pet rabbits, ferrets and four-toed hedgehogs in Japan. Methods: We histologically/cytologically investigated 1098 samples from 883 rabbits, 812 samples from 521 ferrets and 561 samples from 468 four-toed hedgehogs that were collected at Miwa Exotic Animal Hospital and submitted to the Laboratory of Veterinary Pathology, The University of Tokyo between 2006 and 2020. The examinations of necropsy samples were not included in the present study. Results: Of the 1098 samples from diseased rabbits, 721 (65.7%) were diagnosed as neoplastic and 377 (34.3%) were as non-neoplastic. Uterine adenocarcinoma (21.1%), cutaneous soft tissue sarcoma (15.1%) and mammary gland adenocarcinoma (8.2%) were the most commonly encountered neoplasms. Endometrial hyperplasia (35.8%), testicular atrophy (4.2%) and uterine adenomyosis (3.7%) were the most common non-neoplastic lesions in rabbits. Of the 812 samples from diseased ferrets, 487 (60.0%) were diagnosed as neoplastic and 325 (40.0%) were as non-neoplastic. Adrenocortical tumor (23.2%), lymphoma (19.3%) and pancreatic islet cell tumor (11.5%) were the most commonly encountered neoplasms. Extramedullary hematopoiesis in the spleen (10.5%), lymph node hyperplasia (7.7%) and cholangiohepatitis (4.6%) were the most common non-neoplastic lesions in ferrets. Of the 561 samples from diseased hedgehogs, 338 (60.2%) were diagnosed as neoplastic and 223 (39.8%) were as non-neoplastic. Endometrial stromal tumor and endometrial mixed tumor (23.7%), oral squamous cell carcinoma (13.0%) and cutaneous soft tissue sarcoma (11.5%) were the most commonly encountered neoplasms. Gingivitis/stomatitis (39.5%), endometrial hyperplasia (18.8%) and dermatitis (6.7%) were the most common non-neoplastic lesions in hedgehogs. Conclusions and clinical relevance: Information obtained from the present study will provide a useful reference for veterinarians working with these exotic animals. Based on the literature search, this is the largest-scale retrospective study on disease incidence in hedgehogs.

Prolactin has a rich mechanistic set of actions and signaling in order to elicit developmental effects in mammals. Historically, prolactin has been appreciated as an endocrine peptide hormone that is responsible for final, functional mammary gland development and lactation. Multiple signaling pathways impacted upon by the microenvironment contribute to cell function and differentiation. Endocrine, autocrine and paracrine signaling are now apparent in not only mammary development, but also in cancer, and involve multiple cell types including those of the immune system. Multiple ligands agonists are capable of binding to the prolactin receptor, potentially expanding receptor function. Prolactin has an important role not only in tumorigenesis of the breast, but also in a number of hormonally responsive cancers such as prostate, ovarian and endometrial cancer, as well as pancreatic and lung cancer. Although pituitary and extra-pituitary sources of prolactin such as the epithelium are important, stromal sourced prolactin is now also being recognized as an important factor in tumor progression, all of which potentially signal to multiple cell types in the tumor microenvironment. While prolactin has important roles in milk production including calcium and bone homeostasis, in the disease state it can also affect bone homeostasis. Prolactin also impacts metastatic cancer of the breast to modulate the bone microenvironment and promote bone damage. Prolactin has a fascinating contribution in both physiologic and pathologic settings of mammals.

Our understanding of the molecular events underpinning the development of mammalian organ systems has been increasing rapidly in recent years. With the advent of new and improved next-generation sequencing methods, we are now able to dig deeper than ever before into the genomic and epigenomic events that play critical roles in determining the fates of stem and progenitor cells during the development of an embryo into an adult. In this review, we detail and discuss the genes and pathways that are involved in mammary gland development, from embryogenesis, through maturation into an adult gland, to the role of pregnancy signals in directing the terminal maturation of the mammary gland into a milk producing organ that can nurture the offspring. We also provide an overview of the latest research in the single-cell genomics of mammary gland development, which may help us to understand the lineage commitment of mammary stem cells (MaSCs) into luminal or basal epithelial cells that constitute the mammary gland. Finally, we summarize the use of 3D organoid cultures as a model system to study the molecular events during mammary gland development. Our increased investigation of the molecular requirements for normal mammary gland development will advance the discovery of targets to predict breast cancer risk and the development of new breast cancer therapies.

Embryonic mammary gland development involves the formation of mammary placodes, invagination of flask-shaped mammary buds and development of miniature bi-layered ductal trees. Currently there is a good understanding of the factors that contribute to ectodermal cell movements to create these appendages and of pathways that lead to mammary specification and commitment. Gene expression profiles of early bipotent mammary stem cells populations as well as cell surface proteins and transcription factors that promote the emergence of unipotent progenitors have been identified. Analyses of these populations has illuminated not only embryonic mammary development, but highlighted parallel processes in breast cancer. Here we provide an overview of the highly conserved pathways that shape the embryonic mammary gland. Understanding the dynamic signaling events that occur during normal mammary development holds considerable promise to advance attempts to eliminate cancer by restoring differentiative signals.

Sheep, cows, cats, and rabbits are kept by humans for agricultural purposes and as companion animals. Much of the mammary research in these species has focussed on mastitis in the case of ruminants and rabbits, and mammary tumourigenesis in cats and rabbits. However, similarities with the human breast suggest that these species may be currently underutilised as valuable comparative models of breast development and disease. The mammary gland undergoes cyclical postnatal development that will be considered here in the context of these non-traditional model species, with a focus on the mammary microenvironment at different postnatal developmental stages. The second part of this review will consider mammary tumour development. Ruminants are thought to be relatively 'resistant' to mammary tumourigenesis, likely due to multiple factors including functional properties of ruminant mammary stem/progenitor cells, diet, and/or the fact that production animals undergo a first parity soon after puberty. By contrast, unneutered female cats and rabbits have a propensity to develop mammary neoplasms, and subsets of these may constitute valuable comparative models of breast cancer.

Emerging evidence has shown that several SOX family transcription factors are key regulators of stem/progenitor cell fates in the mammary gland. These cell-fate regulators are often upregulated in breast cancer and contribute to tumor initiation and progression. They induce lineage plasticity and the epithelial-mesenchymal transition, which promotes tumor invasion, metastasis, and therapeutic resistance. SOX factors act through modulating multiple oncogenic signaling pathways in breast cancer. In addition to the cell-autonomous functions, new evidence suggests they can shape the tumor immune microenvironment. Here, we will review the molecular and functional evidence linking SOX factors with mammary gland development and discuss how these cell-fate regulators are co-opted in breast cancer.

The accumulation of 131-iodine is known to occur in the extrathyroidal tissues, such as the salivary glands and the gastrointestinal tract during radioactive iodine therapy. It is considered to be due to the expression of sodium iodide symporter. A 36-year-old lactating female was treated for remnant ablation for papillary thyroid carcinoma (pT1bN1bM0 pStage I) on the condition that she refrained from breastfeeding and was separated from her child and was milked to promote the excretion of 131-iodine until the accumulation disappeared on scintigram imaging. Scintigram showed the decrease and disappearance of the 131-iodine accumulation and the external dose rate was measured and confirmed to have decreased on the day 7 and 14 after the treatment. In conclusion, remnant ablation may be available by milking and separation from the child for 7 to 14 days.

Purpose Computed tomographic (CT) scans in adolescents have increased dramatically in recent years. However, the effects of cumulative low-dose exposures on the development of radiation sensitive organs, such as the mammary gland, is unknown. The purpose of this work was to define the effects of dose rate on mammary organ formation during puberty, an especially sensitive window in mammary development. We used a fractionated low-dose x-ray exposure to mimic multiple higher dose CT scans, and we hypothesized that fractionated exposure would have less of an effect on the number of mammary gland defects compared with an acute exposure. Methods and Materials Female mice were subjected to fractionated low-dose x-ray exposure (10 cGy/d for 5 days), acute x-ray exposure (1 × 50 cGy), or sham exposure. As the wide genetic diversity in humans can play a role in a person’s response to irradiation, 2 genetically diverse mouse strains differing in radiation sensitivity (BALB/c-sensitive; C57BL/6-resistant) were used to investigate the role of genetic background on the magnitude of the effect. Results Unexpectedly, our data reveal that multiple low-dose exposures produce greater immune and mammary defects for weeks after exposure compared with controls. The most pronounced defects being increased ductal branching in both strains and a greater percentage of terminal end buds in the BALB/c strain of mice exposed to fractionated radiation compared with sham. Radiation-induced defects near the terminal end bud were also increased in both strains. Conclusions The findings suggest that fractionated low-dose exposures are potentially more damaging to organ development compared with an equivalent, single acute exposure and that genetic background is an important parameter modifying the severity of these effects.

Mammary epithelial cells synthesize and secrete norepinephrine (NE) into breast milk to regulate β-casein expression through the adrenergic β2 receptor. We investigated the expression, localization, and roles of NE transporter (NET) in the mammary epithelium during lactation. mRNA and protein levels of NET were determined in primary normal human mammary epithelial cells (pHMECs) and non-malignant human mammary epithelial MCF-12A cells. In nursing CD1 mice, NET localized to the apical membranes of the mammary epithelium. The intracellular NE content of pHMECs incubated with NE increased. Although the β-casein concentration in milk was slightly higher at day 10 than at day 2 of lactation, the NE concentration and lactation-related proteins were only slightly changed on days 2-10. Restraint stress increased the NE concentration in milk from nursing mice and NET protein levels were significantly higher than in non-stressed nursing mice. NET is expressed on the apical membrane of mammary epithelial cells and incorporates NE in milk into cells, potentially regulating the NE concentration in milk.

Pro-inflammatory conditions have long been associated with mammary carcinogenesis and breast cancer progression. The underlying mechanisms are incompletely understood but signaling of pro-inflammatory cytokine TNFα through its receptors TNFR1 and TNFR2 is a major mediator of inflammation in both obesity and in the response of tissues to radiation, 2 known risk factors for the development of breast cancer. Here, we demonstrated the loss of one TNFR2 allele led to ductal hyperplasia in the mammary gland with increased numbers of mammary epithelial stem cell and terminal end buds. Furthermore, loss of one TNFR2 allele increased the incidence of breast cancer in MMTV-Wnt1 mice and resulted in tumors with a more aggressive phenotype and metastatic potential. The underlying mechanisms include a preferential activation of canonical NF-κB signaling pathway and autocrine production of TNFα. Analysis of the TCGA dataset indicated inferior overall survival for patients with down-regulated TNFR2 expression. These findings unravel the imbalances in TNFR signaling promote the development and progression of breast cancer, indicating that selective agonists of TNFR2 could potentially modulate the risk for breast cancer in high-risk populations.

Summary The mammary epithelium undergoes several rounds of extensive proliferation during the female reproductive cycle. Its expansion is a tightly regulated process, fueled by the mammary stem cells and these cells' unique property of self-renewal. Sufficient new cells have to be produced to maintain the integrity of a tissue, but excessive proliferation resulting in tumorigenesis needs to be prevented. Three well-known tumor suppressors, p53, p16INK4a, and p19ARF, have been connected to the limiting of stem cell self-renewal and proliferation. Here we investigate the roles of these three proteins in the regulation of self-renewal and proliferation of mammary epithelial cells. Using mammary epithelial-specific mouse models targeting Trp53 and Cdkn2a, the gene coding for p16INK4a and p19ARF, we demonstrate that p53, p16INK4a, and p19ARF do not play a significant role in the limitation of normal mammary epithelium self-renewal and proliferation, whereas in the presence of the inflammatory cytokine TNF-α, Trp53−/−Cdkn2a−/− mammary basal cells exhibit amplified proliferation., Highlights • p53, p16INK4a, and p19ARF do not limit self-renewal of mammary epithelial cells • p53, p16INK4a, and p19ARF do not limit proliferation of mammary epithelial cells • TNF-α stimulates mammary basal cell organoid formation and proliferation • Trp53−/− Cdkn2a−/− organoids are sensitized to TNF-α-induced proliferation, Van Weele et al. demonstrate that combined deficiency of the tumor suppressors p53, p16INK4a, and p19ARF does not limit self-renewal and proliferation in normal murine mammary epithelial cells. They show that the inflammatory cytokine TNF-α promotes organoid formation and proliferation and sensitizes Trp53−/−Cdkn2a−/− organoids to excessive proliferation. These findings indicate that during inflammation, p53, p16INK4a, and p19ARF may limit proliferation.

MicroRNAs (miRNAs) are small non-coding RNAs that are involved in mammary gland development and lactation in livestock. Little is known about the roles of miRNAs in ovine mammary gland development, hence in this study the expression profiles of miRNAs of the mammary gland tissues of ewes at peak-lactation and during the non-lactating period were investigated using RNA sequencing. A total of 147 mature miRNAs were expressed in the two periods. Compared with peak-lactation, eight miRNAs in the non-lactating ewe mammary gland were significantly up-regulated, whereas fifteen miRNAs were down-regulated. A KEGG analysis revealed that the target genes of the up-regulated miRNAs were significantly enriched in lysosome, Wnt and MAPK signaling pathways, while the target genes of down-regulated miRNAs were significantly enriched in the PI3K-Akt signaling pathway, protein processing in endoplasmic reticulum and axon guidance. These results suggest that further study of the differentially expressed miRNAs could provide a better understanding of the molecular mechanisms of mammary development and lactation in sheep.

Purpose of the study The purpose of this study was to assess the potential for improvement of routine breast and organs at risk contouring in patients treated in the lateral decubitus position using Workflow Box™ (Mirada Medical™, UK) automatic contouring software. Materials and methods Automatic contouring of the breast by this software is currently based on an atlas created from isodoses representing 95% of the prescribed dose in a population of patients previously treated at institut Curie. Forty cases of breast cancer (20 right breasts and 20 left breasts) were contoured by three radiation oncologists specialized in breast cancer, allowing the creation of a new atlas in the automatic contouring software. This study assessed the quality of contouring in 20 patients (ten right breasts and ten left breasts) by comparing manual contouring performed by the expert radiation oncologists (reference) with those generated by the old and new atlases developed at Institut Curie. The accuracy of contouring was assessed by overlap volume and the associated standard deviations. Results Breast contouring based on the new atlas and by radiation oncologists presented a mean overlap of 0.80 ± 0.09 for the right breast and 0.81 ± 0.06 for the left breast. By comparing volumes of interest contoured by radiation oncologists and those obtained from the old atlas, mean overlap volumes were 0.75 ± 0.08 and 0.74 ± 0.13 for the right and left breasts, respectively. Twenty cases (ten right breasts and ten left breasts) used to create the new atlas were also reprocessed by this same atlas in order to assess the quality of automatic contouring of the breast when the case was already known to the software. The mean overlap volume in this case was 0.84 ± 0.08 for the right breast and 0.83 ± 0.06 for the left breast. Finally, after automatic contouring of organs at risk by means of the new atlas, the mean overlap volume was 0.87 ± 0.04 for the heart and 0.93 for each lung (±0.05 for the right lung and ± 0.04 for the left lung). Conclusion Workflow Box™ automatic contouring software, based on our new atlas provides reliable and clinically relevant organs at risk and breast contouring. The contours proposed by the software from the new atlas were better than those obtained with the previous atlas based on 95% isodoses obtained from old treatment plans. This software has therefore become more efficient, justifying its use in routine clinical practice for breast cancer contouring in patients treated in the lateral decubitus position. Investigations are currently underway to develop a fully automated process to ensure reliable, robust and operator-independent contouring and breast cancer treatment dosimetry in the lateral decubitus position. Promising preliminary results have already been obtained.

Progesterone (Pg) is a pregnancy-related hormone that prepares the endometrium for the implantation of the fertilized zygote and suppresses myometrial contractility for the maintenance of pregnancy. At high concentrations, it acts as a natural immunosuppressant avoiding the rejection of a half allogeneic foetus. It is the precursor of many other related steroid hormones, but what is its role in the human breast? In this chapter, we will discuss some aspects related to Pg and its role in breast development and in the neoplastic disease. Understanding the mechanisms related to Pginduced effects in the normal and neoplastic mammary gland will light the way to exploit this hormone signalling pathway therapeutically. We will introduce some aspects of the effects of progestins in normal breast development, breast cancer risk and in neoplastic growth, and we will describe ongoing clinical trials in breast cancer using progestins or antiprogestins. Fil: Lamb, Caroline Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Fabris, Victoria Teresa. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Lanari, Claudia Lee Malvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina

The mammary gland is a highly dynamic tissue that undergoes repeated cycles of growth and involution during pregnancy and menstruation. It is also the site from which breast cancers emerge. Organoids provide an invitro model that preserves several of the cellular, structural, and microenvironmental features that dictate mammary gland function invivo and have greatly advanced our understanding of glandular biology. Their tractability for genetic manipulation, live imaging, and high throughput screening have facilitated investigation into the mechanisms of glandular morphogenesis, structural maintenance, tumor progression, and invasion. Opportunities remain to enhance cellular and structural complexity of mammary organoid models, including incorporating additional cell types and hormone signaling.

ASTRACT Mastitis an inflammation of mammary gland in dairy herds is a key concern of economic losses worldwide caused by bacteria and its toxins. In this study, we investigated pro-inflammatory and anti-inflammatory cytokines (IL-2, IL-1β, IL-6, TGF-β, IL-10, TNF-α), lactoferrin and albumin, and milk composition in normal dairy cows and lactating cows with symptoms of sub-clinical and clinical mastitis. Lactating cows with clinical mastitis showed marked increase in IL-2, IL-6, IL-1β, TNF-α and a decrease in anti-inflammatory levels of TGF-β, IL-10, milk parameters fat, protein, SNF, lactose and increase in pH compared to normal lactating cows. Subclinical lactating cows showed significant alteration in TNF-α, IL-1β when compared to normal lactating cows. There was no significant difference between IL and 2 and IL-6 in normal and subclinical lactating cows. The subclinical cows also did not exhibit significant difference in TGF-β, albumin, milk fat, protein and pH when compared with normal lactating cows. Findings in the present study indicate that cytokines together with proteins lactoferrin and albumin can be considered as prospective markers in early detection of subclinical and clinical mastitis.

To investigate the role of adipose tissue in reproductive function and mammary gland development and function, we have examined lipodystrophic (LD) mice. LD mice of both sexes are sterile, but fertility can be restored with leptin injections. Mammary glands from lipodystrophic mice were rudimentary and lacked terminal end buds. Leptin-injected LD mice were able to become pregnant, showed normal pregnancy-associated glandular proliferation despite a smaller glandular area, were able to produce a small amount of milk that had grossly normal content of milk proteins and neutral lipids, but could not sustain pups to weaning. In order to separate the individual requirements for 1) adipokines such as leptin, 2) estradiol, and 3) physical epithelial-adipocyte interactions, we performed a series of experiments with both lipodystrophic mice and ob (obese mice with a mutation in the lep gene encoding the adipokine leptin) mice that received either estradiol treatment or preadipocyte transplant. The resulting fat pad did not rescue the defect in mammary gland development in lipodystrophic mice. The defect also could not be rescued with estradiol pellets. Ob/ob mice, like LD mice, lack leptin and estradiol, but retain adipose tissue. Ob mice have defective mammary gland development. However, in striking contrast to what was observed in lipodystrophic mice, reconstitution of a WT fat pad in ob mice rescued the defect in mammary gland development. Estradiol treatment did not rescue mammary gland development in ob mice. Therefore direct interaction between mammary gland epithelia and adipocytes is a requirement for full invasion and expansion of the gland, but is not required for glandular proliferation during pregnancy and milk production.

Feeding management of the postnatal and preweaning calf has an important impact on calf growth and development during this critical period and affects the health and well-being of the calves. After birth, an immediate and sufficient colostrum supply is a prerequisite for successful calf rearing. Colostrum provides high amounts of nutrient as well as non-nutrient factors that promote the immune system and intestinal maturation of the calf. The maturation and function of the neonatal intestine enable the calf to digest and absorb the nutrients provided by colostrum and milk. Therefore, colostrum intake supports the start of anabolic processes in several tissues, stimulating postnatal body growth and organ development. After the colostrum feeding period, an intensive milk feeding protocol, that is, at least 20% of BW milk intake/day, is required to realise the calf potential for growth and organ development during the preweaning period. Insufficient milk intake delays postnatal growth and may have detrimental effects on organ development, for example, the intestine and the mammary gland. The somatotropic axis as the main postnatal endocrine regulatory system for body growth is stimulated by the intake of high amounts of colostrum and milk and indicates the promotion of anabolic metabolism in calves. The development of the forestomach is an important issue during the preweaning period in calves, and forestomach maturation is best achieved by solid feed intake. Unfortunately, intensive milk-feeding programmes compromise solid feed intake during the first weeks of life. In the more natural situation for beef calves, when milk and solid feed intake occurs at the same time, calves benefit from the high milk intake as evidenced by enhanced body growth and organ maturation without impaired forestomach development during weaning. To realise an intensive milk-feeding programme, it is recommended that the weaning process should not start too early and that solid feed intake should be at a high extent despite intensive milk feeding. A feeding concept based on intensive milk feeding prevents hunger and abnormal behaviour of the calves and fits the principles of animal welfare during preweaning calf rearing. Studies on milk performance in dairy cows indicate that feeding management during early calf rearing influences lifetime performance. Therefore, an intensive milk-feeding programme affects immediate as well as long-term performance, probably by programming metabolic pathways during the preweaning period.

Understanding how progenitor cell function is regulated in the mammary gland is an important developmental problem that has significant implications for breast cancer. Although it had been assumed that the expression the α6β4 integrin (β4) is restricted to the basal lineage, we report that alveolar progenitor cells in the mouse mammary gland also express this integrin based on analysis of single cell RNA-Seq data. Subsequent experiments using a mouse mammary epithelial cell line (NMuMG) confirmed this finding and revealed that β4 is essential for maintaining progenitor function as assessed by serial passage mammosphere assays. These data were substantiated by analyzing the alveolar progenitor population isolated from nulliparous mouse mammary glands. Based on the finding that the alveolar progenitor cells express Whey Acidic Protein (WAP), WAP-Cre mice were crossed with itgβ4(flox/flox) mice to generate conditional knock-out of β4 in alveolar progenitor cells. These itgβ4(flox/flox) WAP-Cre(+) mice exhibited significant defects in alveologenesis and milk production during pregnancy compared to itgβ4(flox/flox) WAP-Cre(−) mice, establishing a novel role for the β4 integrin in alveolar progenitor function and alveologenesis.

Objective of the paper is to describe mechanisms leading to mammary involution in ewes and changes taking place in the mammary gland during that period. Mammary involution is the regression of mammary tissue to non-secreting state and takes place as initiated, gradual or senile. In mutton-type production systems cessation of lactation is abrupt, whilst in dairy-type production systems it is progressive or abrupt. The period from cessation of milk removal until the beginning of subsequent lactation period is termed ‘dry-period’ and is distinguished into stage of active involution, stage of the ‘steady-state’ involution and stage of redevelopment and lactogenesis. The ‘dry-period’ is important in health management of sheep for optimum milk production during the subsequent lactation period, as it is necessary for renewal of mammary epithelial cells. Mammary involution is influenced by decreased activity of galactopoietic hormones and local mechanisms in response to milk accumulation in the gland. Milk accumulation plays a major role in triggering apoptosis of epithelial cells. There are two pathways leading to apoptosis, an intrinsic and an extrinsic. During involution, significant histological changes take place in the mammary gland, mainly reduction (up to 3.6% up to the 4th day) of the epithelial area of the gland and increase of its stromal part. The mammary gland is particularly susceptible to infections after cessation of lactation; progressively, that changes; a keratin plug is formed at the teat orifice, leucocytes accumulate in the gland and concentrations of immunoglobulins and lactoferrin increase. Udder health management at the end of a lactation period aims to cure infections that have occurred during the previous lactation period and prevent new intramammary infections during the ‘dry-period’. Culling ewes with at least one mammary gland permanently damaged or ones chronically affected or others with incidents of relapsing mastitis or not fully respondent to mastitis treatment during the preceding lactation period contributes to decrease of veterinary expenses for mastitis control in the flock, elimination of sources of potential infection for other animals in the flock and decrease of flock bulk somatic cell counts in the subsequent lactation period. Culling should be complemented with intramammary antibiotic administration at the end of a lactation period. In many clinical studies from around the world, administration of antimicrobial agents at the end of a lactation period has been found beneficial. Antibiotic administration at drying-off may be performed to all animals in a flock (‘complete’) or only to those considered to be infected (‘selective’). In all cases, maintenance of the prescribed withdrawal periods is essential to safeguard public health. The procedure should always be applied as part of a strategic udder health management plan in a flock; implementation improves the welfare of animals and affords significant financial benefits to the farmer. Correct udder health management in ewes at the end of a lactation period will contribute to improved mammary health for the forthcoming lactation period.

This experiment was conducted to investigate the effects of heat stress (HS) on the feed intake, milk production and composition and metabolic alterations in the mammary gland of dairy cows. Twenty Holstein cows were randomly assigned to one of two treatments according to a completely randomized design. Half of the cows were allocated to the HS group in August (summer season), and the other half were assigned to the HS-free group in November (autumn season). HS reduced (P

New insights into the host-microbiota relationship have recently emerged with the advancement of molecular technologies such as next-generation sequencing. This article presents the current knowledge regarding the interaction between bacteria and the immune system of the gut, the uterus, and the mammary gland of cattle.

Milk lactose, pH, somatic cell count (SCC), bacterial count and NaCl are indirect markers of the mammary gland status; they can be used to screen milk quality and its technological properties. The variability of milk composition and coagulation properties from 1272 individual goat milk samples as a function of the udder health indicators was investigated. High lactose concentrations were associated with reduced coagulation time, but weakened curd firmness traits. High pH values impaired coagulation, while high SCC had delayed coagulation time but reduced curd-firming rate. Samples with high bacterial count were characterised by softer curds, but had faster curd-firming and larger syneresis rates. High concentrations of NaCl were associated with reduced fat, protein, and casein content, and impaired coagulation traits. These results show that the concurrent analysis of these markers can be highly informative and suitable to monitor the quality of milk destined for cheese-making.

Dairy cows experience a negative energy balance due to increasing energy demands and insufficient voluntary feed intake in the transition from late pregnancy to early lactation. For supplying sufficient energy toward the conceptus and the mammary gland, insulin sensitivity in peripheral tissues is reduced leading to adipose tissue mobilization. Adiponectin, an insulin-sensitizing adipokine, is presumably related to energy metabolism and could play an important role in these metabolic adaptations. We hypothesize (1) that primiparous cows would differ from pluriparous cows in their circulating adiponectin concentrations during the transition from late pregnancy to early lactation and (2) that feeding different energy levels would affect the adiponectin concentrations during early lactation in dairy cows. For the first hypothesis, we examined 201 primiparous and 456 pluriparous Holstein dairy cows on three experimental farms. Ante partum, primiparous cows had lower adiponectin and greater NEFA concentrations than pluriparous cows, but vice versa post partum. Hence, adiponectin might be involved in the energy partitioning in primiparous cows (conceptus and lactation vs other still growing body tissues) with changing priorities from pregnancy to lactation. For the second hypothesis, 110 primiparous and 558 pluriparous Holstein and Simmental dairy cows in six experimental farms received either roughage with 6.1 or 6.5 MJ NEl/kg dry matter (adjusted with different amounts of wheat straw) ad libitum, combined with either 150 or 250 g concentrates/kg energy corrected milk. Greater amounts of concentrate lead to greater milk yield, but did not affect the blood variables. The higher energy level in the roughage led to greater glucose and IGF-1 but lower adiponectin in pluriparous cows. Further studies are needed to elucidate the mechanisms behind the roughage effect and its metabolic consequences.

Made available in DSpace on 2019-10-06T16:42:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-10-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) This study aimed to evaluate the association of CD3+, CD4+, and CD8+ T cells and tumor-infiltrating macrophages (TIMs) with the clinical parameters of female dogs harboring mammary gland tumors. Thirty female dogs affected with mammary carcinomas were used, and all tumors were histologically classified as complex carcinoma and were triple-negative phenotype determined by immunohistochemistry. Freshly frozen sections were used to determine CD3+, CD4+ and CD8+ T cells by immunohistochemistry, and TIMs were determined by immunofluorescence assays. Ten out of the 30 dogs showed lymph node metastasis at diagnosis. Fifteen dogs had a tumor of grade I (15/30), nine (9/30) had a tumor of grade II and six (6/30) had a tumor of grade III. The mean overall survival was 680.5 days (± 200.4). Dogs with sentinel lymph node positivity (10/30) (P =.0035) and dogs that developed metastasis (P =.0001) showed a shorter survival time. In addition, dogs with a high level of inflammatory infiltrate in tumor tissues presented a shorter survival time (P =.0001) than that of other dogs. Dogs with tumors containing higher numbers of CD3+ T cells (P =.001), CD4+ T cells (P =.001), or TIM cells (P

Functional RNAs, such as microRNAs, are emerging as innovative tools in the treatment of aggressive and incurable cancers. In this study, we explore the potential of silica dioxide nanoparticles (SiO2NPs) in the delivery of biologically active miRNAs. Focusing on the tumor-suppressor miR-34a, we evaluated miRNAs delivery by SiO2NPs into the mammary gland, using in vitro as well as in vivo model systems. We showed that silica nanoparticles can efficiently deliver miR-34a into normal and cancer epithelial cells grown in culture without major signs of toxicity. Delivered miRNA retained the ability to silence artificial as well endogenous targets and can reduce the growth of mammospheres in 3D culture. Finally, miR-34a delivery through intra-tumor administration of SiO2NPs leads to a reduced mammary tumor growth. In conclusion, our studies suggest that silica nanoparticles can mediate the delivery of miR-34a directly into mammary tumors while preserving its molecular and biological activity.

Postmortem examination of an 8-year-old intact female sugar glider euthanized for end-stage neoplasia revealed a large mass in the mammary gland, masses in the liver, and a few small masses in the lung. Histologic evaluation of the lesions revealed hepatocellular carcinoma, mammary gland adenocarcinoma, and pulmonary granulomata associated with a trematode larva. Ribosomal RNA gene sequencing identified a diplomastid fluke, Neodiplostomum americanum. This parasite utilizes snails as first intermediate hosts and birds of prey as definitive hosts, but the complete life cycle is not known; the hosts of the intervening life stages had not been previously reported. To the authors’ knowledge, this is the first reported case of N. americanum in a sugar glider.

The review reports recent knowledge on the role and significance of somatic cells in sheep mammary gland physiology. Somatic cell count represents an index of animal health status and it is considered an elective standard for the milk quality detection. However, there is still the need to deepen the knowledge on this complex parameter and to define a cut-off value of SCC in sheep milk in order to guarantee safety and quality of sheep milk and to sustain the development of dairy sheep productive system. Quantitative limits and detection methods for somatic cell count in sheep milk production are discussed in relation to qualitative, hygienic and technological aspects. Furthermore somatic cell level and qualitative patterns in sheep milk are discussed in relation to milk cytokines, endogenous proteolytic enzymes and pathogenic bacteria; this new approach to milk quality evaluation is reported to highlight possible early and reliable indicators able to predict and ensure quality and safety of sheep milk and dairy products. Finally, the role of mammary epithelial cells in the defence of the mammary gland represents a new research field, which still needs to be explored in sheep milk.

A 5-year-old intact female rabbit was presented with bilateral ocular lesions. In both eyes, a fleshy mass was emerging from the iris. Focal corneal edema, neovascularization, generalized rubeosis iridis, +1 aqueous flare, and dyscoria could also be identified. In addition, hepatomegaly was detected by clinical examination. Hematology and serum biochemistry results included leukocytosis, slight anemia, hyperproteinemia, and mild increased liver enzyme activities. Abdominal ultrasound revealed a moderate enlarged liver with a heterogeneous structure as well as ascites. Thoracic radiography showed generalized pulmonary metastasis. By request of the owner, the rabbit was euthanized. Pathological examination revealed nodules composed of malignant epithelial cells in the lung, anterior and posterior uvea of both eyes, pericardium, diaphragm, liver, mammary gland, and the left ovary. A carcinoma of the mammary gland was suspected as the primary tumor. To our knowledge, this is the first report of a carcinoma with bilateral uveal metastasis in a rabbit.

Background Based on estrogen active substances, many women consume soy foods in the belief that it could prevent breast cancer (BC). Women with different molecular subtypes would be likely to have diverse reactions to soy foods, especially those with the estrogen-receptor-positive (ER+) subtype. The aim of the current study is to identify the differentially expressed genes (DEGs) on soy foods in premenopausal patients with Lumina A subtype of BC (LABC) after soy food treatment, and to further investigate the critical molecule change. Materials and Methods GSE58792 retrieved from Gene Expression Omnibus was analyzed to obtain DEGs using GEO2R. Gene Ontology and pathway enrichment analysis were performed using FunRich and GeneMINIA. Overall survival of critical genes was performed by the Kaplan-Meier plotter online tool. Results A total of 108 DEGs were obtained from the dataset, among which 35 were up-regulated and 73 down-regulated. Soy foods significantly reduced the expression of TFF3, TFF1, GATA3, and ESR1, which were related to the activity of the ER-related pathway and the sensitivity of tamoxifen. Furthermore, the lower expressions of TOX3, FSIP1, ESR1, and CLGN were related to prolonged survival time of patients with BC. The most significant signaling pathways were epithelial-to-mesenchymal transition in up-regulated DEGs, mesenchymal-to-epithelial transition, and mammary gland alveolus development in down-regulated DEGs, which were all related to the development and prognosis of BC. Conclusions Soy foods could dramatically alter the ER-related gene profile in LABC. Particularly, down-regulated DEGs of TFF3, TFF1, GATA3, and ESR1 might weaken the sensitivity of tamoxifen and increase the efficacy of neoadjuvant chemotherapy in premenopausal patients with LABC.

The intricate architecture of branched tissues and organs has fascinated scientists and engineers for centuries. Yet-despite their ubiquity-the biophysical and biochemical mechanisms by which tissues and organs undergo branching morphogenesis remain unclear. With the advent of three-dimensional (3D) culture models, an increasingly powerful and diverse set of tools are available for investigating the development and remodeling of branched tissues and organs. In this review, we discuss the application of 3D culture models for studying branching morphogenesis of the mammary gland and the mammalian lung in the context of normal development and disease. While current 3D culture models lack the cellular and molecular complexity observed in vivo, we emphasize how these models can be used to answer targeted questions about branching morphogenesis. We highlight the specific advantages and limitations of using 3D culture models to study the dynamics and mechanisms of branching in the mammary gland and mammalian lung. Finally, we discuss potential directions for future research and propose strategies for engineering the next generation of 3D culture models for studying tissue morphogenesis.

Long term high-concentrate (HC) diet feeding induces subacute ruminal acidosis (SARA), which is reported to trigger a pro-inflammatory response. This study aimed to investigate the role of nucleotide-binding oligomerization domain protein 1 (NOD1) in initiating the pro-inflammatory response triggered by grain-induced SARA in the mammary gland of mid-lactating dairy cows. Twelve multiparous mid-lactating Holstein cows (455 ± 28 kg) were randomly assigned into two groups to conduct the experiment for 18 weeks as follows: one group was fed a low-concentrate (LC) diet as a control (40% grain), and the other was fed an HC diet as a treatment (60% grain). Overall, the results showed that a decreased rumen pH and elevated γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP) concentrations in the HC group compared with LC group. The concentration of pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6 and tumour necrosis factor-alpha (TNF-α), significantly increased in the lacteal vein of the HC group than LC group. The mRNA expression levels of NOD1, receptor-interacting protein2 (RIP2), NF-κBp65 (p65), IL-1β, IL-6, IL-8 and TNF-α, which involved in inflammatory response, were up-regulated in the HC-induced mammary gland. The changes of the target proteins, including NOD1, p65 and pp65 presented the same tendency as those of the target genes. Collectively, long-term high concentrate feeding-induced SARA increased the rumen iE-DAP concentration which activated NOD1-NF-κB signalling pathway-dependent inflammation in the mammary gland of mid-lactating cows.

Progesterone (Pg)/progesterone receptor (PR) signaling drives mammary gland side-branching and alveologenesis, but the mechanisms through which Pg/PR signaling functions remain to be clarified. Using in vitro and in vivo models and histological and molecular analyses, we determined the role of Zfhx3 transcription factor in mammary gland development driven by Pg/PR signaling. Postnatal deletion of Zfhx3 in mouse mammary epithelial cells attenuated side-branching morphogenesis and alveologenesis. These effects were undetectable in the absence of Pg/PR signaling. During the estrus cycle, Zfhx3 expression corresponded to that of Pg, being at the highest level at the diestrus stage; Zfhx3 deletion inhibited mammary gland branching more potently at diestrus than estrus stage. Loss of Zfhx3 not only attenuated the expansion of stem/progenitor cells driven by Pg/PR signaling, but also impaired the function of Pg/PR signaling in the transcriptional activation of multiple genes. In addition, Pg/PR signaling significantly expanded PR- and Zfhx3-positive epithelial cells, and induced the physical association of ZFHX3 with PR. These findings establish Zfhx3 as an integral transcription factor of Pg/PR signaling in driving side-branching and alveologenesis during mammary gland development.

Introduction Breast cancer is the most frequent cancer in women worldwide. The primary treatment is breast-conserving surgery or mastectomy with an adequate clearance margin. Diathermy blade is used extensively in breast-conserving surgery. Surgical smoke produced as a side product has cancer-specific molecular features. Differential mobility spectrometry (DMS) is a rapid and affordable technology for analysis of complex gas mixtures. In our study we examined surgical smoke from malignant and benign breast tissue created with a diathermy blade using DMS. Material and methods Punch biopsies of 4 mm diameter from breast cancer surgical specimens were taken during gross dissection of fresh surgical specimen and placed in a well plate. The measurement system is a custom-built device called automatic tissue analysis system (ATAS) based on a DMS sensor. Each specimen was incised with a diathermy blade and the surgical smoke was analyzed. Results We examined 106 carcinoma samples from 21 malignant breast tumors. Benign samples (n = 198) included macroscopically normal mammary gland (n = 82), adipose tissue (n = 88) and vascular tissue (n = 28). The classification accuracy when comparing malignant samples to all benign samples was 87%. The sensitivity was 80% and the specificity was 90%. The classification accuracy of carcinomas to ductal and lobular was 94%, 47%, respectively. Conclusions Benign and malignant breast tissue can be identified with ATAS. These results lay foundation for intraoperative margin assessment with DMS from surgical smoke.

The main function of the mammary gland is to produce milk which contains the crucial active components for infant development. The process of milk production is tightly controlled at a local level by hormones, growth factors, noncoding RNAs and other factors. Circular RNAs (CircRNAs) are specifically expressed across tissues or developmental stages where they have potential regulatory functions. Competing endogenous RNAs (ceRNAs), sharing miRNA recognition elements (MREs), emerge to compete for miRNA binding and, in turn, regulate each other. Although circRNAs have been determined in rat and bovine mammary glands during lactation, these studies simply investigated the expression of circRNAs in the mammary gland with no exploration of ceRNAs of circRNA-mRNA. In the current study, mRNA, miRNA and circRNA were sequenced in goat mammary tissue at 2 stages during lactation (early and mature). All data (mRNA, miRNA, and circRNA) were co-expressed together from the same samples. ceRNAs (mRNA-miRNA-circRNA) were found to be involved in goat lactation. Therefore the findings of the current investigation suggest a novel approach through the application of a ceRNA network (mRNA-miRNA-circRNA) for the analysis of gene functionality in biological processes such as lactation.

Human milk is a dynamic protein-protease system that delivers bioactive peptides to infants. The pH of milk changes from the mother’s mammary gland to the infant’s digestive tract. Although the release of human milk peptides has been studied during in vivo or in vitro digestion, these models did not explicitly vary nor observe the effect of pH. The objective of this research was to determine the effect of pH on the proteolysis of human milk. Using high-resolution accurate-mass Orbitrap mass spectrometry, profiles of endogenous human milk peptides before and after incubation at various pH levels have been mapped. Over 5000 peptides were identified. Comparative analyses classified 74 peptides that were consistently found independent of pH alterations, and 8 peptides that were released only at pH 4 or 5 (typical infant gastric pH). Results documented that the proteolysis of milk proteins, particularly β-casein, polymeric immunoglobulin receptor, and α-lactalbumin, is pH-dependent.

Twelve quarters of six lactating cows were inoculated with Mycoplasma leachii strain GN407 through intramammary ductal infusion; another 12 quarters were inoculated with heat-inactivated M. leachii culture medium as negative controls. Multidisciplinary procedures, including clinical assessment, etiology assessment, pathology and immunohistochemistry (IHC), were used to elucidate the pathogenicity of M. leachii in bovine mastitis. From post-inoculation days (PIDs) 3 to 9, 12 inoculated quarters developed mild to severe clinical mastitis and mammary tissue histopathological changes, including inflammatory cell infiltration and architectural destruction of mammary gland ducts. The M. leachii antigen was also detected by IHC in the mammary gland epithelial cells of the inoculated quarters as a weak signal on PID 6 and as a strong signal on PID 9. The control quarters also developed mild mastitis and histopathological changes on PID 9, and M. leachii was also detected by IHC. Throughout the experimental period, the quarters of the negative control cow were clinically and pathologically normal, and the M. leachii antigen was not detected. In conclusion, direct histological and immunohistochemical evidence confirmed that M. leachii causes clinical bovine mastitis through histopathological lesions induced by invasion of the pathogen into mammary gland cells and through inflammatory cell infiltration.

Mammary development takes place during the growing and gestation periods in swine but it also continues after farrowing. In fact, a significant proportion of mammary accretion occurs during lactation and is stimulated by suckling. After piglets are weaned, there is involution of the mammary glands and the process of mammogenesis starts again during the next parity. Suckling of a teat for the first 12 to 14 h after farrowing is not sufficient to maintain lactation, and mammary involution accompanied by alterations in gene transcription will take place. The involution process is reversible within 1 day postpartum but is not reversible if a mammary gland is unsuckled for 3 days. Mammary glands that undergo involution early in lactation do not show further involution in the post-weaning period. The action of a teat being suckled does not only affect mammary development in the ongoing lactation but it also impacts mammogenesis in the following lactation. Indeed, when a mammary gland is not suckled in first parity it has a diminished development and lower milk yield in second parity. Furthermore, it was shown that suckling of a teat for only the first 2 days postpartum in primiparous sows is sufficient to ensure optimal mammary development and milk yield from that teat in the next lactation. The behavior of nursing piglets in early lactation is also affected by whether or not a teat was previously used. Such knowledge on lactation biology is essential in order to develop the best adapted management strategies for the currently used hyperprolific sow lines and to optimize growth rate of their piglets. This review gives an update on the role of suckling for mammary development in lactating sows and on how it can affect management strategies of primiparous sows.

Historically, pre-pubertal development of the bovine mammary gland (MG) has received little attention compared to later development. Recent evidence suggests not only that this period represents a very active time in the development of the MG but also that the first 90 days of life can partially dictate future productivity of the lactating cow. The MG, often considered quiescent during early life (first 3 months), is now known to increase in size by over 60-fold in the same period. The importance of sex steroids in MG development is well classified, but a complex signaling network exists among estrogen, progesterone and other growth factors and hormones. Complicating our understanding of this developmental period further is the discovery that pre-weaning nutrition of the calf not only influences the growth of the mammary parenchyma but may also alter the way in which it responds to mammogenic stimuli. Recent data suggest that feeding calves a higher plane of nutrition improves the ability of the mammary epithelium to respond to estradiol and also alters the way in which the mammary parenchyma and fat pad communicate. It is clear that early life nutrition, although able to influence the MG, is still poorly understood mechanistically. For example, additional evidence suggests that increased feeding rates in early life alter the morphology of myoepithelial cells in the mammary epithelium. Further data have also suggested a role for other cell types, such as immune cells, in the penetration of the mammary parenchyma into the fat pad during the early life development of the MG suggesting that mammary development is not only controlled by the local tissue population (parenchyma and fat pad) but perhaps systemically by other tissue types (i.e., immune system). Understanding the roles of these various stimuli and signaling pathways as they relate to the development of the MG in early life may hold the key to unlocking the potential for the optimal development of this crucial organ and, in turn, may lead to improvements in other phases of mammary development and milk yield potential.

Summary DACH1 abundance is reduced in human malignancies, including breast cancer. Herein DACH1 was detected among multipotent fetal mammary stem cells in the embryo, among mixed lineage precursors, and in adult basal cells and (ERα+) luminal progenitors. Dach1 gene deletion at 6 weeks in transgenic mice reduced ductal branching, reduced the proportion of mammary basal cells (Lin− CD24med CD29high) and reduced abundance of basal cytokeratin 5, whereas DACH1 overexpression induced ductal branching, increased Gata3 and Notch1, and expanded mammosphere formation in LA-7 breast cells. Mammary gland-transforming growth factor β (TGF-β) activity, known to reduce ductal branching and to reduce the basal cell population, increased upon Dach1 deletion, associated with increased SMAD phosphorylation. Association of the scaffold protein Smad anchor for receptor activation with Smad2/3, which facilitates TGF-β activation, was reduced by endogenous DACH1. DACH1 increases basal cells, enhances ductal formation and restrains TGF-β activity in vivo., Highlights • Dach1 is expressed in mammary gland fetal stem cells and adult luminal cells • Dach1 expands mammary gland basal/myoepithelial cells • Dach1 induces post-natal mammary gland ductal formation • Dach1 retrains TGF-β activity in the mammary gland in vivo, DACH1 abundance, is reduced in human malignancies, including breast cancer. In this article, Dr. Pestell and his colleagues showed that DACH1 is expressed in multipotent mammary gland fetal stem cells and both the adult basal and ERα+ luminal cells, promotes mammary gland stem cell and basal/myoepithelial cell expansion, promotes mammary gland ductal branching, and restrains TGF-β action in the pubertal mammary gland.

Canonical WNT/β- catenin signaling plays pivotal roles in mammary development and tumorigenesis; and aberrant activation of this pathway is frequently observed in human breast cancer, correlating with poor outcome. However, the mechanisms underlying WNT-mediated mammary tumorigenesis remain incompletely understood. Here, we used mouse mammary tumor virus (MMTV)-Wnt1 transgenic mice, which develop aggressive mammary adenomas, to examine whether Limb-Bud-and-Heart (LBH) - a WNT/β-catenin target transcription co-factor overexpressed in human triple-negative breast cancers with WNT pathway hyperactivation, contributes to WNT-induced tumorigenesis. We found LBH is specifically overexpressed in basal epithelial tumor cells of MMTV-WNT1 mammary tumors reminiscent of its’ basal cell-restricted expression pattern in the normal postnatal mammary gland. To determine the role of LBH in mammary tumorigenesis, we crossed MMTV-Wnt1 mice with epithelial-specific Keratin 14/K14-Cre;Lbh(loxP) knockout mice. Mammary glands from virgin LBH-deficient MMTV-Wnt1 mice exhibited reduced hyperplasia, cell proliferation and increased apoptosis. Importantly, LBH inactivation in mammary epithelium significantly delayed tumor onset in MMTV-Wnt1 transgenic mice, with a median tumor-free survival of 32.5 weeks compared to 22.5 weeks in control LBH wild type MMTV-Wnt1 mice (p

The mammary tissue is characterized by its capacity to adapt in response to a wide variety of changing conditions. This adaptation capacity is referred to as the plasticity of mammary tissue. In dairy ruminants, lactation is challenged by modifications that can either be induced on purpose, such as by modifying management practices, or occur involuntarily, when adverse environmental constraints arise. These modifications can elicit both immediate changes in milk yield and composition and carryover effects that persist after the end of the challenge. This review focuses on the current knowledge concerning the cellular mechanisms underlying mammary tissue plasticity. The main mechanisms contributing to this phenomenon are changes in the activity and number of mammary epithelial cells (MECs). Changes in the number of these cells result from variations in the rates of cell proliferation and death as well as changes in the rate MEC exfoliation. The number of MECs also depends on the number of resident adult mammary stem cells and their progenitors, which can regenerate the pools of the various mammary cells. Several challenges, including changes in milking frequency, changes in level of feed supply and hormonal manipulations, have been shown to modulate milk yield together with changes in mammary cell activity, turnover and exfoliation. Epigenetic changes may be an additional mechanism of adaptation. Indeed, changes in DNA methylation and reductions in milk yield have been observed during once-daily milking and during mastitis in dairy cows and may affect cell activity persistently. In contrast to what has been assumed for a long time, no carryover effect on milk yield were observed after feed supply challenges in dairy cows and modification of milking frequency in dairy goats, even though the number of mammary cells was affected. In addition, mammary tissue plasticity has been shown to be influenced by the stage of lactation, health status and genetic factors. In conclusion, the cellular mechanisms underlying mammary tissue plasticity are diverse, and the mammary tissue either does or does not show elastic properties (with no permanent deformation), in response to environmental changes.

The present study aimed to evaluate the mechanisms modulated by dietary arginine supplementation to sows during lactation regarding antioxidant capacity and vascularization of mammary glands. At 109 days of gestation, animals were transferred to individual farrowing crates equipped with manual feeders and automatic drinker bowls. Environmental temperature and humidity inside the farrowing rooms were registered every 15 min. At farrowing, sows were assigned in a completely randomized design to a control diet (CON) or the CON diet supplemented with 1.0% L-arginine (ARG). A total of three gilts and two sows were fed the CON diet, whereas three gilts and three sows were fed ARG diets. Sows were fed a fixed amount of 6.0 kg/day, subdivided equally in four delivery times (0700, 1000, 1300 and 1600 h) for 21 days. At weaning, sows were slaughtered and mammary tissue samples and blood from the pudendal vein were collected. Data were analyzed considering each sow as an experimental unit. Differences were considered at Plt;0.05. L-arginine fed sows presented lower messenger RNA (mRNA) expression for prolactin receptor (P=0.002), angiopoietin1 (P=0.03) and receptor tyrosine kinase (P=0.01); higher mRNA expression for prostaglandin synthase 1 (P=0.01); a trend of decrease for glucocorticoid receptor (P=0.06) and IGF receptor 1 (P=0.07); and a trend (P=0.05) for an increased glutathione peroxidase mRNA expression. The angiopoietin2:angiopoietin1 mRNA ratio tended to increase (P=0.07) in ARG fed sows. L-arginine fed sows had greater (P=0.04) volumetric proportion of blood vessels and a trend of enhance (P=0.07) in the number of blood vessels per mm2. These findings show that 1.0% ARG supplementation to sows activates proliferative mechanisms, may improve mammary tissues' angiogenesis and tended to increase mRNA expression of genes that encode antioxidant enzymes in mammary gland of sows.