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PURPOSE Immunohistochemistry (IHC) is a newer technique for assessing the estrogen receptor (ER) status of breast cancers, with the potential to overcome many of the shortcomings associated with the traditional ligand-binding assay (LBA). The purpose of this study was to evaluate the ability of ER status determination by IHC, compared with LBA, to predict clinical outcome—especially response to adjuvant endocrine therapy—in a large number of patients with long-term clinical follow-up. PATIENTS AND METHODS ER status was evaluated in 1,982 primary breast cancers by IHC on formalin-fixed paraffin-embedded tissue sections, using antibody 6F11 and standard methodology. Slides were scored on a scale representing the estimated proportion and intensity of positive-staining tumor cells (range, 0 to 8). Results were compared with ER values obtained by the LBA in the same tumors and to clinical outcome. RESULTS An IHC score of greater than 2 (corresponding to as few as 1% to 10% weakly positive cells) was used to define ER positivity on the basis of a univariate cut-point analysis of all possible scores and disease-free survival (DFS) in patients receiving any adjuvant endocrine therapy. Using this definition, 71% of all tumors were determined to be ER-positive by IHC, and the level of agreement with the LBA was 86%. In multivariate analyses of patients receiving adjuvant endocrine therapy alone, ER status determined by IHC was better than that determined by the LBA at predicting improved DFS (hazard ratios/ P = 0.474/.0008 and 0.707/.3214, respectively) and equivalent at predicting overall survival (0.379/.0001 and 0.381/.0003, respectively). CONCLUSION IHC is superior to the LBA for assessing ER status in primary breast cancer because it is easier, safer, and less expensive, and has an equivalent or better ability to predict response to adjuvant endocrine therapy.

Staphylococcus aureus is a major pathogen causing intramammary infection and mastitis in dairy cows. S. aureus genotypes (GT) can differ significantly in their ability to diffuse and persist in the herd; while the association of virulence gene carriage with epidemiological behavior remains unclear, a role for secreted proteins has been postulated. We characterized the secretome of six S. aureus strains belonging to two genotypes with opposite within-herd prevalence, GTB (high) and GTS (low), corresponding to sequence types (ST) 8 and 398, by high-resolution tandem mass spectrometry and differential analysis with Proteome Discoverer. Data are available via ProteomeXchange with identifier PXD029571. Out of 720 identified proteins, 98 were unique or more abundant in GTB/ST8 and 68 in GTS/ST398. GTB/ST8 released more immunoglobulinbinding proteins, complement and antimicrobial peptide inhibitors, enterotoxins, and metabolic enzymes, while GTS/ST398 released more leukocidins, hemolysins, lipases, and peptidases. Furthermore, GTB/ST8 released the von Willebrand factor protein, staphylokinase, and clumping factor B, while GTS released the staphylococcal coagulase and clumping factor A. Hence, GTB/ST8 secretomes indicated a higher propensity for immune evasion and chronicity and GTS/ST398 secretomes for cellular damage and inflammation, consistent with their epidemiological characteristics. Accordingly, GTS/ST398 secretions were significantly more cytotoxic against bovine PBMCs in vitro. Our findings confirm the crucial role of extracellular virulence factors in S. aureus pathogenesis and highlight the need to investigate their differential release adding to gene carriage for a better understanding of the relationship of S. aureus genotypes with epidemiological behavior and, possibly, disease severity.

In 2010 the worldwide population of domesticated ruminants was estimated to be 3.6 billion [1]. Many of these animals are used for milk production or as suckler animals where the offspring are consumed for meat. These functions are dependent on the lactational capacity of the female ruminant, thus postnatal mammary development impacts future lactational productivity [2, 3] whilst mastitis is a major welfare issue that also causes financial losses [4].

Studies of microbiota in normal canine milk from healthy dams are sparse. As is the case with blood and urine, it was considered that milk contains no microbiota. Any discovery of bacteria in canine milk is, therefore, often noted to be a result of contamination during sampling or interpreted as mastitis and treated with antibiotics. Milk was collected twice within 19 days after natural parturition from 11 lactating dams, with no general or local clinical signs of mastitis or other disease. The skin and teats were prepared with an antimicrobial protocol prior to each milk sampling. In total, 210 milk samples were collected and assessed for a number of bacterial colonies grown on each plate. Bacterial growth was detected in 180 samples (86%). Staphylococcus pseudintermedius, Enterococcus spp., Clostridium spp., Coagulase-Negative Staphylococci (CoNS), Streptococcus spp., Streptococcus canis, Bacillus spp., Pasteurella spp., and Escherichia coli were identified from pure and/or mixed bacterial growth, listed in descending order of occurrence. Despite the small sample size, the consistent occurrence of bacteria in early postpartum dams indicates a genuine occurrence of bacteria in canine milk, rather than random contamination. The finding of bacteria in the milk of dams should not, therefore, be the sole argument for the diagnosis of mastitis.

Membrane transporters and ion channels that play an indispensable role in metabolite trafficking have evolved to operate in Earth’s gravity. Dysregulation of the transportome expression profile at normogravity not only affects homeostasis along with drug uptake and distribution but also plays a key role in the pathogenesis of diverse localized to systemic diseases including cancer. The profound physiological and biochemical perturbations experienced by astronauts during space expeditions are well-documented. However, there is a paucity of information on the effect of the space environment on the transportome profile at an organ level. Thus, the goal of this study was to analyze the effect of spaceflight on ion channels and membrane substrate transporter genes in the periparturient rat mammary gland. Comparative gene expression analysis revealed an upregulation (p < 0.01) of amino acid, Ca2+, K+, Na+, Zn2+, Cl−, PO43−, glucose, citrate, pyruvate, succinate, cholesterol, and water transporter genes in rats exposed to spaceflight. Genes associated with the trafficking of proton-coupled amino acids, Mg2+, Fe2+, voltage-gated K+-Na+, cation-coupled chloride, as well as Na+/Ca2+ and ATP-Mg/Pi exchangers were suppressed (p < 0.01) in these spaceflight-exposed rats. These findings suggest that an altered transportome profile contributes to the metabolic modulations observed in the rats exposed to the space environment.

The objective of this study was to assess the transcriptome of the mammary tissue of four yaks during the whole lactation cycle. For this purpose, biopsies of the mammary gland were performed at −30, −15, 1, 15, 30, 60, 120, 180, and 240 days relative to parturition (d). The transcriptome analysis was performed using a commercial bovine microarray platform and the results were analyzed using several bioinformatic tools. The statistical analysis using an overall false discovery rate ≤ 0.05 for the effect of whole lactation and p < 0.05 for each comparison identified >6000 differentially expressed genes (DEGs) throughout lactation, with a large number of DEGs observed at the onset (1 d vs. −15 d) and at the end of lactation (240 d vs. 180 d). Bioinformatics analysis revealed a major role of genes associated with BTA3, BTA4, BTA6, BTA9, BTA14, and BTA28 in lactation. Functional analysis of DEG underlined an overall induction of lipid metabolism, suggesting an increase in triglycerides synthesis, likely regulated by PPAR signaling. The same analysis revealed an induction of amino acid metabolism and secretion of protein, with a concomitant decrease in proteasome, indicating a major role of amino acid handling and reduced protein degradation in the synthesis and secretion of milk proteins. Glycan biosynthesis was induced for both N-glycan and O-glycan, suggesting increased glycan content in the milk. The cell cycle and immune response, especially antigen processing and presentation, were strongly inhibited during lactation, suggesting that morphological changes are minimized during lactation, while the mammary gland prevents immune hyper-response. Transcripts associated with response to radiation and low oxygen were enriched in the down-regulated DEG affected by the stage of lactation. Except for this last finding, the functions affected by the transcriptomic adaptation to lactation in mammary tissue of yak are very similar to those observed in dairy cows.

The mammary gland is a vital exocrine organ that has evolved in mammals to secrete milk and provide nutrition to ensure the growth and survival of the neonate The mouse mammary gland displays extraordinary plasticity each time the female undergoes pregnancy and lactation, including a sophisticated process of tertiary branching and alveologenesis to form a branched epithelial tree and subsequently milk-producing alveoli. Upon the cessation of lactation, the gland remodels back to a simple ductal architecture via highly regulated involution processes. At the cellular level, the plasticity is characterised by proliferation of mammary cell populations, differentiation and apoptosis, accompanied by major changes in cell function and morphology. The mammary epithelium requires a specific stromal environment to grow, known as the mammary fat pad. Mammary adipocytes are one of the most prominent cell types in the fat pad, but despite their vast proportion in the tissue and their crucial interaction with epithelial cells, their physiology remains largely unknown. Over the past decade, the need to understand the properties and contribution of mammary adipocytes has become more recognised. However, the development of adequate methods and protocols to study this cellular niche is still lagging, partially due to their fragile nature, the difficulty of isolating them, the lack of reliable cell surface markers and the heterogenous environment in this tissue, which differs from other adipocyte depots. Here, we describe a new rapid and simple flow cytometry protocol specifically designed for the analysis and isolation of mouse mammary adipocytes across mammary gland developmental stages.

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.

Phenylalanine (PHE) and to a lesser extent TYR are two commonly used amino acid tracers for measuring protein metabolism in a variety of species and tissues. The model examined in this paper was developed to resolve trans-organ and stable isotope dilution data collected from experiments with lactating dairy cows using these tracers. Two methods of solving the model, i.e. as two four-pool submodels, one representing PHE and the other TYR, or as an integrated eight-pool model, are investigated and the alternative solutions are contrasted. Solving the model as the two four-pool submodels rather than the integrated 8-pool model is preferred as the equations are slightly simpler and their application less susceptible to any compounding of measurement errors. The data used to illustrate the model were taken from experiments conducted to investigate the effects of high and low protein diets on the partitioning of PHE and TYR between milk protein synthesis and other metabolic fates by the mammary gland.

Background Inflammation of the mammary tissue (mastitis) is one of the most detrimental health conditions in dairy ruminants and is considered the most economically important infectious disease of the dairy sector. Improving mastitis resistance is becoming an important goal in dairy ruminant breeding programmes. However, mastitis resistance is a complex trait and identification of mastitis-associated alleles in livestock is difficult. Currently, the only applicable approach to identify candidate loci for complex traits in large farm animals is to combine different information that supports the functionality of the identified genomic regions with respect to a complex trait. Methods To identify the most promising candidate loci for mastitis resistance we integrated heterogeneous data from multiple sources and compiled the information into a comprehensive database of mastitis-associated candidate loci. Mastitis-associated candidate genes reported in association, expression, and mouse model studies were collected by searching the relevant literature and databases. The collected data were integrated into a single database, screened for overlaps, and used for gene set enrichment analysis. Results The database contains candidate genes from association and expression studies and relevant transgenic mouse models. The 2448 collected candidate loci are evenly distributed across bovine chromosomes. Data integration and analysis revealed overlaps between different studies and/or with mastitis-associated QTL, revealing promising candidate genes for mastitis resistance. Conclusion Mastitis resistance is a complex trait influenced by numerous alleles. Based on the number of independent studies, we were able to prioritise candidate genes and propose a list of the 22 most promising. To our knowledge this is the most comprehensive database of mastitis associated candidate genes and could be helpful in selecting genes for functional validation studies.

Malic Enzyme 1 (ME1) supports lipogenesis, cholesterol synthesis, and cellular redox potential by catalyzing the decarboxylation of L-malate to pyruvate, and the concomitant reduction of NADP to NADPH. We examined the contribution of ME1 to the development of obesity by provision of an obesogenic diet to C57BL/6 wild type (WT) and MOD-1 (lack ME1 protein) female mice. Adiposity, serum hormone levels, and adipose, mammary gland, liver, and small intestine gene expression patterns were compared between experimental groups after 10 weeks on a diet. Relative to WT female mice, MOD-1 female mice exhibited lower body weights and less adiposity; decreased concentrations of insulin, leptin, and estrogen; higher concentrations of adiponectin and progesterone; smaller-sized mammary gland adipocytes; and reduced hepatosteatosis. MOD-1 mice had diminished expression of Lep gene in abdominal fat; Lep, Pparg, Klf9, and Acaca genes in mammary glands; Pparg and Cdkn1a genes in liver; and Tlr9 and Ffar3 genes in the small intestine. By contrast, liver expression of Cdkn2a and Lepr genes was augmented in MOD-1, relative to WT mice. Results document an integrative role for ME1 in development of female obesity, suggest novel linkages with specific pathways/genes, and further support the therapeutic targeting of ME1 for obesity, diabetes, and fatty liver disease.

Breast milk is a complex biological fluid. Additionally to its nutritional impact, it contains diverse bioactive elements related to early metabolic programming and molecular structures, such as microRNA related to the epigenetic signaling process. Now, we know that human milk is not sterile and contains a significant diversity of microorganisms such as bacteria (bacterioma), viruses (viroma) and fungi (mycobiome), all of which integrate the concept of the human milk microbiota. Apparently the origin of this microbiota is found in the entero-mammary circulation, as well as in the retrograde circulation of the skin of the mammary gland, although it is speculated in what percentage the microbiota of the oral cavity of the infant contributes in a significant way. From a functional point of view, it has been shown the transfer of this microbiota to the infant's digestive tract, which is related to better digestive tolerance, lower frequency and intensity of dysfunctionalities of the brain-intestine-microbiota axis, and improved immunity, among others.La leche materna es un fluido biológico complejo, ya que además de nutrir, contiene diversos elementos bioactivos relacionados con el fenómeno de programación metabólica temprana e incluso contiene estructuras moleculares relacionadas con el proceso de señalización epigenética tales como los microARN. Ahora sabemos que la leche humana no es estéril y contiene una diversidad significativa de microorganismos tales como bacterias (bacterioma), virus (viroma) y hongos (micobioma), todo lo cual integra el concepto de microbiota de leche humana. Al parecer el origen de esta microbiota se encuentra en la circulación enteromamaria, así como en la circulación retrógrada de la piel de glándula mamaria, aunque se especula en qué porcentaje la microbiota de la cavidad bucal del lactante contribuye de forma significativa. Desde el punto de vista funcional, se ha demostrado transferencia de esta microbiota hacia el tubo digestivo del lactante, lo que se relaciona con una mejor tolerancia digestiva, menor frecuencia e intensidad de disfuncionalidades del eje cerebro-intestino-microbiota, y mejora de la inmunidad, entre otras.

Adult stem cells like mammary and mesenchymal stem cells have received significant attention because these stem cells possess therapeutic potential in treating many animal diseases. These cells can be administered in an autologous or allogenic fashion, either freshly isolated from the donor tissue or previously cultured and expanded in vitro. The expansion of adult stem cells is a prerequisite before therapeutic application because sufficient numbers are required in dosage calculation. Stem cells directly and indirectly (by secreting various growth factors and angiogenic factors called secretome) act to repair and regenerate injured tissues. Recent studies on mammary stem cells showed in vivo and in vitro expansion ability by removing the blockage of asymmetrical cell division. Compounds like purine analogs (xanthosine, xanthine, and inosine) or hormones (progesterone and bST) help increase stem cell population by promoting cell division. Such methodology of enhancing stem cell number, either in vivo or in vitro, may help in preclinical studies for translational research like treating diseases such as mastitis. The application of mesenchymal stem cells has also been shown to benefit mammary gland health due to the ‘homing’ property of stem cells. In addition to that, the multiple positive effects of stem cell secretome are on mammary tissue; healing and killing bacteria is novel in the production of quality milk. This systematic review discusses some of the studies on stem cells that have been useful in increasing the stem cell population and increasing mammary stem/progenitor cells. Finally, we provide insights into how enhancing mammary stem cell population could potentially increase terminally differentiated cells, ultimately leading to more milk production.

Nuclear magnetic resonance spectroscopy was applied to investigate the association between milk metabolome and udder quarter health status in dairy cows. Mammary gland health status was defined by combining information provided by traditional somatic cell count (SCC) and differential SCC (DSCC), which expresses the percentage of neutrophils and lymphocytes over total SCC. Quarter milk samples were collected in triplicate (d 1 to 3) from 10 Simmental cows, 5 defined as cases and 5 defined as controls according to SCC levels at d 0. A total of 120 samples were collected and analyzed for bacteriology, milk composition, SCC, DSCC, and milk metabolome. Bacteriological analysis revealed the presence of mostly coagulase-negative staphylococci in quarter milk samples of cows defined as cases. Nuclear magnetic resonance spectra of all quarter samples were first analyzed using the unsupervised multivariate approach principal component analysis, which revealed a specific metabolomic fingerprint of each cow. Then, the supervised cross-validated orthogonal projections to latent structures discriminant analysis unquestionably showed that each cow could be very well identified according to its milk metabolomic fingerprint (accuracy = 95.8%). The comparison of 12 different models, built on bucketed 1-dimensional NOESY spectra (noesygppr1d, Bruker BioSpin) using different SCC and DSCC thresholds, corroborated the assumption of improved udder health status classification ability by joining information provided by both SCC and DSCC. Univariate analysis performed on the 34 quantitated metabolites revealed lower levels of riboflavin, galactose, galactose-1-phosphate, dimethylsulfone, carnitine, hippurate, orotate, lecithin, succinate, glucose, and lactose, and greater levels of lactate, phenylalanine, choline, acetate, O-acetylcarnitine, 2-oxoglutarate, and valine, in milk samples with high somatic cells. In the 5 cases, results of the udder quarter with the highest SCC compared with its symmetrical relative were in line with quarter-level findings. Our study suggests that increased SCC is associated with changes in milk metabolite fingerprint and highlights the potential use of different metabolites as novel indicators of udder health status and milk quality.

Breast cancer is the most common type of cancer in the world. The aggressive and invasive nature of these tumors is explicable by the presence of cancer stem cells, which have self-renewal and initiation potentials that are analogous to the capabilities seen in normal mammary stem cells. The characterization of a protein’s role in a normal mammary gland, along with how this function is altered in cancerous tissue, enables enhanced understanding of the potential for specific novel drug targets. Transcription factor EB (TFEB) is currently known for its role as a master lysosomal regulator, and it has been shown to assist with cancerous progression through metabolic reprogramming. Yet, the key functions served by TFEB in mammary acini formation and its effects on mammary stem cells have never been determined. I have shown that the silencing of TFEB interrupts and disorganizes the development of mammary acini, unlike mere transcriptional inactivation. Furthermore, TFEB was not found to directly regulate the differentiation of mammary stem cells into a luminal lineage. Instead, TFEB appears to markedly contribute to the early proliferation of MCF10A cells, but additional investigation is necessary to characterize the mechanism. This work will guide future exploration into TFEB's potential as a drug target.

10 páginas, 1 tabla, 2 figuras., As higher feed efficiency in dairy ruminants means a higher capability to transform feed nutrients into milk and milk components, dierences in feed efficiency are expected to be partly linked to changes in the physiology of the mammary glands. Therefore, this study aimed to determine the biological functions and key regulatory genes associated with feed efficiency in dairy sheep using the milk somatic cell transcriptom, This research was funded by the Junta de Castilla y León (JCyL, Spain; project CSI276P18) and the Spanish Research State Agency (Agencia Estatal de Investigación, Spain; project PID2020-113441RB-I00 and MCIN/AEI/10.13039/501100011033). Cofunding by the European Regional Development Fund (ERDF/FEDER) is also acknowledged. Research activities for the University of Leon were funded by the SMARTER project through the H2020 research and innovation program of the European Commission (Grant Agreement No. 772787).

9 páginas, 3 figuras, 2 tablas., This study evaluated the influence of a temporary nutritional protein restriction (NPR) performed, under commercial conditions, in prepubertal female lambs on first lactation milk production traits and the inflammatory response triggered by an inflammatory challenge of the. From 40 Assaf female lambs, we defined a control group (C n = 20), which received a standard diet for replacement lambs and the NPR group (n = 20), which received the same diet but without soybean meal between 3 and 5 months of age. About 150 days after lambing, 24 of these ewes (13 NPR, 11C) were subjected to an intramammary infusion of E. coli lipopolysaccharide (LPS). Our dynamic study identified indicator traits of local (SCC) and systemic (rectal Ta, IL-6, CXCL8, IL-10, IL-36RA, VEGF-A) response to the LPS challenge. The NPR did not show significant effects on milk production traits and did not affect the SCC and rectal Ta after the LPS challenge. However, the NPR had a significant influence on 8 of the 14 plasma biomarkers analysed, in all the cases with higher relative values in the C group. The effects observed on VEGF-A (involved in vasculogenesis during mammary gland development and vascular permeability) and IL-10 (a regulatory cytokine classically known by its anti-inflammatory action) are the most remarkable to explain the differences found between groups. Whereas further studies should be undertaken to confirm these results, our findings are of interest considering the current concern about the future world's demand for protein and the need for animal production systems to evolve toward sustainability., The nutritional and inflammatory challenge experiments and the sample collection described in this study were funded by the SMARTER project through the Horizon 2020 research and innovation program of the European Commission (Grant Agreement No. 772787). The cytokine/interleukine panel analyses were funded by the Spanish national project EPIMILKSHEEP (Ref. RTI2018–093535-B-I00), funded by the Spanish Ministry of Science and Innovation.

Breast cancer brain metastases remain largely incurable. Although several mouse models have been developed to investigate the genes and mechanisms regulating breast cancer brain metastasis, these models often lack clinical relevance since they require the use of immunocompromised mice and/or are poorly metastatic to brain from the mammary gland. We describe the development and characterisation of an aggressive brain metastatic variant of the 4T1 syngeneic model (4T1Br4) that spontaneously metastasises to multiple organs, but is selectively more metastatic to the brain from the mammary gland than parental 4T1 tumours. As seen by immunohistochemistry, 4T1Br4 tumours and brain metastases display a triple-negative phenotype, consistent with the high propensity of this breast cancer subtype to spread to brain. In vitro assays indicate that 4T1Br4 cells have an enhanced ability to adhere to or migrate across a brain-derived endothelial monolayer and greater invasive response to brain-derived soluble factors compared to 4T1 cells. These properties are likely to contribute to the brain selectivity of 4T1Br4 tumours. Expression profiling and gene set enrichment analyses demonstrate the clinical relevance of the 4T1Br4 model at the transcriptomic level. Pathway analyses implicate tumour-intrinsic immune regulation and vascular interactions in successful brain colonisation, revealing potential therapeutic targets. Evaluation of two histone deacetylase inhibitors, SB939 and 1179.4b, shows partial efficacy against 4T1Br4 metastasis to brain and other sites in vivo, and potent radio-sensitising properties in vitro. The 4T1Br4 model provides a clinically relevant tool for mechanistic studies and to evaluate novel therapies against brain metastasis. This article has an associated First Person interview with Soo-Hyun Kim, joint first author of the paper., Summary: The authors introduce a new syngeneic mouse model of spontaneous breast cancer brain metastasis, demonstrate its phenotypic, functional and transcriptomic relevance to human TNBC brain metastasis, and test novel therapies.

Mammary gland (MG) is a dynamic organ that is essential for the production and secretion of breast milk. During pregnancy the MG undergoes a critical phase of remodeling, which is accompanied by the differentiation of mammary epithelial cells (MECs). During lactation, the MG requires high level of energy for proper folding of proteins in the secretory pathway, which takes place in the endoplasmic reticulum (ER). Limited evidence has been reported on the impact of cannabis or its components, delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD), on the differentiation of MECs. However, both THC and CBD have been reported to induce ER stress in various cell types, resulting in impacting cellular function. Furthermore, consequences of cannabis use in the perinatal period on breast milk composition have not been reported. Using the HC11 cell line, we investigated whether THC and CBD evoke ER stress in MECs resulting in the impaired cellular function. We also reported on the effects of cannabis use during pregnancy and lactation on the levels of macronutrients and bioactive factors in the breast milk of cannabis users. Relative to control, 10μM THC and 10μM CBD reduced mRNA levels of milk proteins (CSN2 and WAP) and lipid synthesizing enzymes (FASN, FABP4, PLIN2 and LPL), as well as whey acidic protein and lipid levels. In addition, 10μM THC, 10μM CBD, and the combination of 10μM THC + 10μM CBD significantly induced the expression of ER stress genes in HC11 cells. Furthermore, in the milk of women who used cannabis during pregnancy and lactation, the levels of secretory immunoglobulin A (SIgA) were decreased, relative to non-users. Altogether, our findings indicate that cannabis use in the perinatal period may have implications on breast milk composition and infant health. Thesis Doctor of Philosophy (Medical Science) Breast milk is crucial to the nutrition and immunity of the newborn. It is produced by the mammary gland (MG), which is composed of mammary epithelial cells (MEC) that play a key role in producing and secreting proteins and factors into the milk. During pregnancy, the MG undergoes remodeling which is accompanied by differentiation of the MECs (change from unspecialized to a specialized stage that allows the MECs to produce milk proteins and other factors). The disruption in MEC differentiation can result in altered milk production and composition. Cannabis is used by women during pregnancy and breastfeeding. In this study, we investigated the impact of cannabinoids on the MEC differentiation and examined the effect of cannabis use in the perinatal period on the breast milk composition in humans. We reported that cannabinoids reduced the differentiation of MECs and maternal cannabis use during breastfeeding reduced levels of an essential immune factor.

Mastitis is one of the most common diseases of dairy cattle worldwide. The aim of this study was to evaluate the incidence of environmental mastitis and the economic impact of mastitis in a selected dairy cattle farm for the period 2019-2022. A total of 596 cases of mastitis were diagnosed, of which 78% were environmental mastitis. While the number of cases of contagious mastitis decreased over the period under review (from 36 % to 6 %), the number of environmental mastitis cases followed the opposite trend. The economic cost per dairy cow ranged from CZK 2 134 to CZK 4 527. The largest item was the loss of milk from treated dairy cows.

Guangtai Shen, 1 Xiaoqiang Yu, 1 Tianyang Sun, 1 Tian Hou, 1 Heping He, 1 Tubuxin Bai, 1 Zhipeng Zhang, 1 Xiaoli Ding, 1 Qingping Li, 2 Ying Liang, 3 Yuhua Ma 4 1Department of Breast and Thyroid Surgery, Inner Mongolia Xing’an League People’s Hospital, Ulanhot, Inner Mongolia, 137400, People’s Republic of China; 2Department of Operating Room, Inner Mongolia Xing’an League People’s Hospital, Ulanhot, Inner Mongolia, 137400, People’s Republic of China; 3Department of Pathology, Inner Mongolia Xing’an League People’s Hospital, Ulanhot, Inner Mongolia, 137400, People’s Republic of China; 4Department of Anesthesiology, Inner Mongolia Xing’an League People’s Hospital, Ulanhot, Inner Mongolia, 137400, People’s Republic of ChinaCorrespondence: Xiaoqiang YuDepartment of Breast and Thyroid Surgery, Inner Mongolia Xing’an League People’s Hospital, Ulanhot, Inner Mongolia, 137400, People’s Republic of ChinaTel/Fax +86 0482-8413761Email yuxq66_dr@163.comObjective: To verify the volume similarity between unilateral mammary gland and autologous omentum in adult females.Methods: A total of 63 patients diagnosed with stage 0–II breast cancer and partial non-lactating multi-fistula mastitis in the breast surgery department of Inner Mongolia Xing’an League People’s Hospital from 2007 to 2020 were enrolled in the study, including 52 cases of stage 0–II breast cancer and 11 cases of non-lactating multi-fistula mastitis. The volume of the resected mammary gland and the omentum were measured by a “soft tissue measuring cylinder” and recorded. The appearance of the reconstructed breast was compared with that of the healthy side. The correlation between unilateral mammary gland volume and autologous omentum volume was analyzed by linear regression.Results: Valid data were obtained for 60 cases. Affected breast size, curve, texture, nipple, and inframammary fold after omentum breast reconstruction were similar and symmetrical to those of the unaffected side. Postoperative complications occurred in most patients; the majority of these (76.67%) involved numbness of the nipple, and other complications were few. Patient satisfaction with postoperative appearance, feel, and movement of the breast, as well as total treatment costs, was over 75.0%. Linear regression analysis indicates a linear relationship between subcutaneous gland volume (x) and autologous omentum volume (y): y = 0.9847x - 1.2132, R 2 = 0.9742.Conclusion: Only when the dissociated pedicled omentum is completely obtained under laparoscopy can the whole subcutaneous residual cavity of the mammary gland be filled to the same volume. This study verifies that the volume of the unilateral mammary gland is similar to that of the autologous omentum in adult females.Keywords: unilateral mammary gland, omentum, volume, similarity, laparoscope, breast reconstruction

Stem-cell containing mammary basal epithelial cells exist in a quasi-mesenchymal transcriptional state characterized by simultaneous expression of typical epithelial genes and typical mesenchymal genes. Whether robust maintenance of such a transcriptional state is required for adult basal stem cells to fuel self-renewal and regeneration remains unclear. In this work, we utilized SMA-CreER to direct efficient basal cell-specific deletion of Ovol2, which encodes a transcription factor that inhibits epithelial-to-mesenchymal transition (EMT), in adult mammary gland. We identified a basal cell-intrinsic role of Ovol2 in promoting epithelial, and suppressing mesenchymal, molecular traits. Interestingly, Ovol2-deficient basal cells display minimal perturbations in their ability to support tissue homeostasis, colony formation, and transplant outgrowth. These findings underscore the ability of adult mammary basal cells to tolerate molecular perturbations associated with altered epithelia-mesenchymal plasticity without drastically compromising their self-renewal potential.

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 Hippo pathway is a highly conserved kinase cascade in mammals with the proteins YAP and TAZ as its most important downstream effectors that shuttle between cytoplasma and nucleus. It has a crucial role in processes such as embryogenesis, organ size control, homeostasis and tissue regeneration, where mechanosensing and/or cell-cell interactions are involved. As the pathway is associated with many essential functions in the body, its dysregulation is related to many diseases. In contrast to human pathology, a PubMed-search on Hippo, YAP/TAZ and companion animals (horse, equine, dog, canine, cat, feline) retrieved few publications. Because of its high level of functional conservation, it is anticipated that also in veterinary sciences aberrant Hippo YAP/TAZ signaling would be implicated in animal pathologies. Publications on Hippo YAP/TAZ in companion animals are mainly in cats and dogs and related to oncology. Here, we emphasize the important role of YAP/TAZ in liver diseases. First the liver has a remarkable regeneration capacity and a strict size control and the liver has a moderate liver cell renewal (homeostasis). The last years numerous papers show the importance of YAP/TAZ in hepatocellular carcinoma (HCC), hepatocyte differentiation and bile duct epithelial (BEC) cell survival. YAP/TAZ signaling is involved in activation of hepatic stellate cells crucial in fibrogenesis. The availability of drugs (e.g. verteporfin) targeting the YAP/TAZ pathway are described as is their potential usage in veterinary medicine. The aim of this overview is to stimulate researchers’ and clinicians’ interest in the potential role of Hippo YAP/TAZ signaling in veterinary medicine.

The immune system plays fundamental roles in the mammary gland, shaping developmental processes and controlling inflammation during infection and cancer. Here we reveal unanticipated heterogeneity in the myeloid cell compartment during development of virgin, pregnant and involuting mouse mammary glands, and in milk. We investigate the functional consequences of individual and compound chemokine receptor deficiency on cell recruitment. Diverse myeloid cell recruitment was also shown in models of sterile inflammation and bacterial infection. Strikingly, we have shown that inflammation and infection can alter the abundance of terminal end buds, a key developmental structure, within the pubertal mammary gland. This previously unknown effect of inflammatory burden during puberty could have important implications for understanding the control of pubertal timing.

Purpose: To develop a method for evaluation of the radiation dose of an embryo/fetus during lymphoscintigraphy (sentinel lymph node mapping) in pregnant patients with breast cancer.Material and methods: Two pregnant women (aged 43 and 30) with breast cancer stage IIA (T2N0M0), during the second trimester of pregnancy. We used a lymphotropic colloidal radiopharmaceutical labeled with 99mTc. To evaluate the radiation dose of an embryo, each patient had 6 individual dosimeters, which were placed around the abdomen using an elastic bandage at equal distances around the abdomen. Additionally, we placed the 7th dosimeter, it was placed near the injection site (under the mammary gland). After installing individual dosimeters, radiocolloid was injected into the affected mammary gland at four points (periareolar). The administered activity of radiopharmaceutical was 32.5 MBq, and 51.5 MBq. Lymphoscintigraphy was performed 1 hour after injection. First patient underwent sector resection of the left breast with SLN biopsy. The second patient underwent right mastectomy with SLN biopsy and breast reconstruction surgery using a tissue expander.Results: Based on the results of the study, the dose rate was calculated, on the basis of which the fetal radiation doses were calculated in both patients. Comparison of the mathematical data of both patients shows that, the calculated and experimental values of radiation exposure to the fetus during the radionuclide study of sentinel lymph nodes practically coincide. The obtained data shows that during pregnancy (280 days) the embryo/fetus will accumulate a natural radiation background dose of 1960 μSv, which is 2 times higher than the dose from the radionuclide study of sentinel lymph nodes. Thus these results verify the safety of SLN biopsy technology in pregnancy.Conclusion: 1. Radionuclide diagnostic studies of pregnant women determine radiation doses to the embryo/fetus that do not cause any radiation-induced effects in the prenatal period, and the probability of the occurrence of stochastic radiation-induced effects is several times lower than the incidence of endogenous cancers. 2. Radionuclide examination of sentinel lymph nodes appears to be safe for the fetus when conducted in pregnant women diagnosed with breast cancer. 3. In Russian Federation this method is used for the first time in pregnant women with diagnosed breast cancer. This technology has not been previously described in Russian literature.

Purpose Accurate identification and assessment of sentinel lymph node (SLN) using noninvasive imaging methods can play a vital role in tumor staging, surgical planning, and prognostic evaluation. In this study, we assessed the efficacy of B7-H3-targeted molecular-ultrasound imaging for the early SLN detection, and characterization in a mouse model of orthotopic breast cancer. Procedures We established a mouse breast cancer model with lymph node metastasis by injecting MAD-MB 231 cells which were engineered to express firefly luciferase reporter gene into the fat pad of the right 4th mammary gland in female BALB/c nude mice. The sole lymph node (LN) close to the tumor was regarded as the SLN for imaging investigation, which included metastatic and non-metastatic SLNs. The LN in the right 4th mammary gland from normal mice was used as normal control (normal mice LN). The commercially available preclinical streptavidin-coated, perfluorocarbon-containing lipid-shelled microbubbles (VisualSonics, Toronto, Canada) were used to generate B7-H3-targeted microbubbles (MBB7-H3) and control microbubbles (MBControl). Then, ultrasound molecular imaging (USMI) was performed using a high-resolution transducer (MS250; center frequency, 21 MHz; Vevo 2100; VisualSonics, Toronto, Canada) after intravenous injection of microbubbles. Results The SLN was clearly detected and located under conventional (B-mode) and contrast-enhanced ultrasonography with microbubble injection. The metastatic SLNs showed a markedly higher signal from B7-H3-targeted microbubbles (MBB7-H3) compared to the non-metastatic SLNs and normal LNs. The metastatic SLN was further confirmed by ex vivo bioluminescence imaging and eventually verified by histological analysis. Conclusions Our findings suggest the potential value of USMI using B7-H3 targeted microbubbles in breast cancer and establish an effective imaging method for the non-invasive detection and characterization of SLN.

The development of a woman's mammary glands occurs during the fertile age. It is during this period that the glands are subject to morpho-functional changes in the form of proliferation and differentiation of the epithelium in the parenchyma and elements of the stroma as response to the action of many stimulating physiological factors of the body. There changes have a tendency to be repeated several times during the woman's reproductive age. There is enough in-depth knowledge about the object of the cyclic effect of sex hormones in the woman's mammary glands. This study will expand knowledge about the important morphological state of the terminal ducts of the mammary glands before pregnancy and childbirth that can be of great importance in determining the histogenesis of possible pathomorphological changes in the gland of a dyshormonal and neoplastic nature. The aim of the study was to investigate the molecular characteristics of the parenchyma of the terminal ducts of the mammary glands of a non-pregnant woman of reproductive age. The study was carried out on autopsy material of 192 micro-incisions of breast tissue samples from 8 women who died of infectious diseases. Karyometry with the definition of the logarithm of the volume of the nuclei of the parenchymal elements in the structures of the terminal ducts was performed; histological, histochemical and immunohistochemical research methods were applied. The study identified histogenetic series of the epithelial lining in the parenchyma of the terminal ducts of the mammary gland; applying immunohistochemical markers enabled cytotyping of the epithelial elements of these ducts, as well revealing certain functions of the terminal ducts of the mammary glands before pregnancy and childbirth.

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.

The effects of dietary supplementation with citrus peel extract (CPE) on milk biochemical parameters, milk bacterial community, and milk metabolites were evaluated. Eight lactating cows were allocated to a replicated 4 × 4 Latin square. Experimental treatments included the control diet (CON), and CON supplemented with CPE at 50 g/d (CPE50), 100 g/d (CPE100), and 150 g/d (CPE150). Supplementing with CPE linearly decreased milk interleukin-6 and malondialdehyde concentrations and linearly increased lysozyme activity and 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Compared with CON, the milk of CPE150 cows had fewer abundances of several opportunistic pathogens and psychrotrophic bacteria, such as Escherichia-Shigella, Sphingobacterium, Alcaligenes, Stenotrophomonas, and Ochrobactrum. Supplementing with CPE significantly altered the metabolic profiling in the milk. The metabolites of flavonoids were enriched in the milk of cows fed CPE150, while some proinflammation compounds were decreased compared with CON. Correlation analysis showed that the change in the bacterial community might partly contribute to the alteration in the expression of milk cytokines. In conclusion, CPE exerts health-promoting effects (e.g., antioxidant, anti-microbial, and anti-inflammatory) in the mammary metabolism of cows due to its flavonoid compounds, which also provide additional value in terms of milk quality improvement.

Milk fat is the foremost nutrient of milk and a vital indicator in evaluating milk quality. Accumulating evidence suggests that microRNAs (miRNAs) are involved in the synthesis of milk fat. The miR-200c is closely related to lipid metabolism, but little is known about its effect on the synthesis of milk fat in MECs of ewes. Herein, the effect of miR-200c on the proliferation of ovine mammary epithelial cells (MECs) and its target relationship with a predicted target gene were investigated. The regulatory effects of miR-200c on the expression of the target genes and the content of triglycerides in ovine MECs were further analyzed. The results revealed that the expression level of miR-200c was differentially expressed in both eight tissues selected during lactation and in mammary gland tissues at different physiological periods. Overexpression of miR-200c inhibited the viability and proliferation of ovine MECs, while inhibition of miR-200c increased cell viability and promoted the proliferation of ovine MECs. Target gene prediction results indicated that miR-200c would bind the 3′UTR region of pantothenate kinase 3 (PANK3). Overexpression of miR-200c reduced the luciferase activity of PANK3, while inhibition of miR-200c increased its luciferase activity. These findings illustrated that miR-200c could directly interact with the target site of the PANK3. It was further found that overexpression of miR-200c reduced the expression levels of PANK3 and, thus, accelerated the synthesis of triglycerides. In contrary, the inhibitor of miR-200c promoted the expression of PANK3 that, thus, inhibited the synthesis of triglycerides in ovine MECs. Together, these findings revealed that miR-200c promotes the triglycerides synthesis in ovine MECs via increasing the lipid synthesis related genes expression by targeting PANK3.

To understand how genes precisely regulate lactation physiological activity and the molecular genetic mechanisms underlying mammary gland involution, this study investigated the transcriptome characteristics of goat mammary gland tissues at the late gestation (LG), early lactation (EL), peak lactation (PL), late lactation (LL), dry period (DP), and involution (IN) stages. A total of 13,083 differentially expressed transcripts were identified by mutual comparison of mammary gland tissues at six developmental stages. Genes related to cell growth, apoptosis, immunity, nutrient transport, synthesis, and metabolism make adaptive transcriptional changes to meet the needs of mammary lactation. Notably, platelet derived growth factor receptor beta (PDGFRB) was screened as a hub gene of the mammary gland developmental network, which is highly expressed during the DP and IN. Overexpression of PDGFRB in vitro could slow down the G1/S phase arrest of goat mammary epithelial cell cycle and promote cell proliferation by regulating the PI3K/Akt signaling pathway. In addition, PDGFRB overexpression can also affect the expression of genes related to apoptosis, matrix metalloproteinase family, and vascular development, which is beneficial to the remodeling of mammary gland tissue during involution. These findings provide new insights into the molecular mechanisms involved in lactation and mammary gland involution.

Breast Milk is an emulsion of fat in a solution of protein, lactose, and inorganic salts secreted by the mother’s mammary gland. Formula milk is cow’s milk whose nutritional composition is changed in such a way that it can be given to babies without giving side effects, cow’s milk itself is a milk of choice for babies who have no family history of allergies. Defecation is the act of a living for or the process of a creature removing solid or half-solid feces or stool originating from the digestive system of living things. This study aims to determine the comparison of the frequency of defecation in infants aged 0-4 months between breast milk and formula milk in the work area Makassar toddopuli health center in 2019. This study uses the Cross-Sectional Study approach which is the type of research in the form of Observation of independent and dependent variables conducted at the same time. Purposive sampling technique. The respondents of this study were 30 people. The observation was carried out for 4 months by recording the expenditure of the baby’s chapters every day. Because the data is not normally distributed, the Tann Pitney statistical test was used and a p-value of 0,000 (p

Background: Postpartum fatigue is a common disorder worldwide and affects both physical and mental functioning. In breastfeeding women, Prolactin (PRL) is not only involved in immunoregulation, but also responsible for lactation. Prolactin levels in women with chronic fatigue are higher than normal, but a chronic fatigue state inhibits postpartum lactation in humans. Objectives: This paper explored the inhibition mechanism of lactation by postpartum fatigue in rats. Methods: Postpartum fatigue models were built by forcing mother rats to stand in water and divided into 3-hour, 9-hour and 15-hour per day fatigue groups according to the underwater time. Mother rats and their offspring were reunited in a dry cage for 90 minutes every 3 hours for feeding. The expression of PRL, PRL receptor (PRLR), Janus Kinase 2 (JAK 2), and Signal transducers and activators of transcription 5 (STAT5) mRNA were analyzed and the microstructure of mammary gland were observed under light and electron microscopy. Results: The expression of pituitary PRL mRNA and its downstream signaling pathway JAK2 and STAT5 mRNA were down-regulated in the severe postpartum fatigue rats. PRL mRNA responses were dose-related to duration of fatigue. The expression of PRLR mRNA increased. Postpartum fatigue led to functional degeneration of mammary gland. The breast lobules were shrunk and the number of alveoli were decreased. Few milk protein granules and fat droplets were observed in the cytoplasm under transmission electron microscope. Conclusion: Postpartum fatigue inhibits the lactation by down-regulating the expression of PRL and PRL-dependent signaling pathway in rats.

Neuroendocrine neoplasms of the breast (Br-NENs) are rare tumors of the mammary gland. Their definition and classification have been a matter of discussion for more than half a century, as they present some degree of overlap with other malignancies in the mammary gland. The recent evolutions in the understanding of the pathogenesis of neuroendocrine neoplasms (NENs) as well as the novel tools in the diagnostic arsenal of the pathologists, and a better correlation with clinical data, have led to improved classification schemes for these entities, beginning from those arising in digestive and thoracic organs. These new classification concepts have been recently expanded to NENs arising in every anatomical site, with the proposal of a common classification framework that has been applied in the 5th edition of the WHO classification of tumors. These concepts include the recognition of two distinct families of NENs (neuroendocrine tumors, NETs, and neuroendocrine carcinomas, NECs), the identification of mixed neuroendocrine/non-neuroendocrine neoplasms, and the application of definite morphological and immunohistochemical criteria for the diagnosis of NENs. The last WHO classification of Br-NENs, however, still seems to leave several unanswered questions and unmet needs in the understanding of diagnostic and clinical features of these entities. This review will critically revise the current classification of Br-NENs, underlining its lights and shadows and focusing on the identification of diagnostic histopathological criteria that can help the univocal recognition of Br-NET, Br-NEC, and Br-MiNEN.

Subclinical mastitis is an insidious disease of the mammary gland that negatively affects the production of the dairy cow. It is necessary to make a diagnosis in order to control them. The main objective is to determine the prevalence of subclinical mastitis and to identify the main risk factors related to the establishment of mammary infections, particularly subclinical mastitis with multi-resistant Staphylococcus in cattle farms in the region of Sidi-Bel-Abbes. The secondary objectives are to describe the characteristics related to the conduct and hygiene of milking that may increase the risk of intramammary infections. A total of 134 cows aged between 1 and 9 years, of Local Breed (n=3), Crossbreed (n= 84) and Holstein Breed (n=47) in different stages of lactation. Milk samples collected from study cows were subjected to California Mastitis Test (CMT) for each udder quarter milk sample. All animals were clinically examined using a standardized questionnaire in an exhaustive manner and completed in real-time, interviewing the staff in charge of the farms and attending all stages of milking. All dairy cows were apparently healthy with clinically sound udder secreting apparently normal milk. The California Mastitis Test (CMT) was used to analyze the raw milk samples collected. The risk factors studied were either animal-related such as age, breed, lactation rank, stage of lactation (Early, Middle, Late) and parity, or environmental related such as season and collection area. Our results indicate that the prevalence of mastitis in crossbred cows (62.68%) and Holstein cows (35.07%) was significantly higher than that of homebred cows (2.23%) with a clear predominance at the age of 1 to 4 years. The result of CMT performed in the present study showed that the prevalence of subclinical mastitis in udder quarters was significantly higher in crossbred cows 204 (30.72%) than in Holstein cows 124 (36.9%) and local cows 8 (2.38%) respectively. The frequency of udder infections by Staphylococcus increases for both primiparous and multiparous women. The analysis of variance shows that the occurrence of subclinical mastitis is influenced by the stage of lactation especially in early lactation compared to other stages (p < 0.001) and parity (p=0.041) in crossbred cows. The Bacteriology was significantly influenced by stage of lactation (p=0.031), breed (p= 0.017) and season (p= 0.005), respectively. The frequency of mammary infections by Staphylococcus is increasing in both primiparous and multiparous cows. By comparing the sampling areas we can deduce that the western area of the region can be considered as a risk indicator compared to the northern area while the other two areas are not. On the other hand, the association of the two factors, i.e. stage of lactation and sampling zone, the risk of the western zone becomes insignificant (p=0.07) while the middle of the stage of lactation persists as a non-risk phase. The present study allowed us to evaluate the frequency of subclinical mastitis and to identify the different bacterial strains of staphylococci responsible. In Conclusion, determining the risk factors for the development of Subclinical Mastitis allows us to establish a control program to improve the health of the udder of dairy cows and thus increase milk production.

Breast cancer, a hormone‐dependent tumour, generally includes four molecular subtypes (luminal A, luminal B, HER2 enriched and triple‐negative) based on oestrogen receptor, progesterone receptor and human epidermal growth factor receptor‐2. Multiple hormones in the body regulate the development of breast cancer. Endocrine therapy is one of the primary treatments for hormone‐receptor‐positive breast cancer, but endocrine resistance is the primary clinical cause of treatment failure. Prolactin (PRL) is a protein hormone secreted by the pituitary gland, mainly promoting mammary gland growth, stimulating and maintaining lactation. Previous studies suggest that high PRL levels can increase the risk of invasive breast cancer in women. The expression levels of PRL and PRLR in breast cancer cells and breast cancer tissues are elevated in most ER+ and ER− tumours. PRL activates downstream signalling pathways and affects endocrine therapy resistance by combining with prolactin receptor (PRLR). In this review, we illustrated and summarized the correlations between endocrine therapy resistance in breast cancer and PRL, as well as the pathophysiological mechanisms and clinical practices. The study on PRL and its receptor would help explore reversing endocrine therapy‐resistance for breast cancer.

The epithelial cells of the mammary gland secrete extracellular nanovesicles known as exosomes that carry and protect microRNAs and other various signaling biomolecules. Milk exosomes are stable du...

The study aim was to determine the expression of genes potentially related to chronic mastitis at the mRNA and protein levels, viz. chemokine C–C motif receptor 1 (CCR1), C–C motif chemokine ligand 2 (CCL2), C–C motif chemokine ligand 5 (CXCL5), tumor necrosis factor α (TNFα), interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 18 (IL-18), in bovine mammary gland parenchyma. The study examines the differences in expression of selected genes between cows with chronic mastitis caused by coagulase-positive (CoPS) or coagulase-negative staphylococci (CoNS) and those with healthy udders (H). Samples were collected from the udder quarters from 40 Polish Holstein–Friesian cows; 54 of these samples were chosen for analysis based on microbiological analysis of milk taken two days before slaughter. They were categorized into three groups: CoPS (N = 27), CoNS (N = 14) and H (N = 13). The RNA expression was analyzed by RT-qPCR and protein concentration by ELISA. No differences in the mRNA levels of seven genes (TNFα, IL-18, CCR1, IL-1β, CCL2, IL-8, IL-6) and four proteins (TNFα, IL-18, CCR1, IL-1β) were identified between the CoPS and H groups. Higher transcript levels of CXCL5 (p ≤ 0.05) gene were noted in CoPS than in H. Compared to H, higher concentrations of IL-8 and CXCL5 (p ≤ 0.05) were observed in CoPS (0.05 p p

Previous studies have demonstrated nonsteroidal antiinflammatory drug treatment in early lactation had a positive impact on whole-lactation milk production in older cows. The objective of this study was to evaluate proliferative, transcriptional, and epigenetic changes in the mammary gland that could explain increased production responses due to nonsteroidal antiinflammatory drug treatment. Sodium salicylate (SAL; 125 g/d) or water (CON) were administered via oral drench to multiparous Holstein cows (n = 8/treatment) once daily for 3 d beginning approximately 24 h after parturition, and mammary tissue was collected on d 1, 4, and 45 postpartum. Day 1 tissue was collected immediately preceding the initial drench, and d 4 tissue was collected 24 h following the final drench. Blood was collected twice weekly and analyzed for plasma glucose, insulin, β-hydroxybutyrate, free fatty acids, and prolactin. Cows were milked twice daily until d 7 of lactation, and thrice daily for the remainder of the study. Total RNA extracted from tissue was deep-sequenced and analyzed for differential gene expression using DESeq2. We detected no treatment effect on milk yield or plasma metabolites through 45 d of lactation; additionally, no change in mammary epithelial cell proliferation was detected when assessed by Ki67 labeling. Comparison of SAL versus CON revealed that only 16 of 18,286 genes were differentially expressed (false discovery rate

Background/aim Primary osteosarcoma of the breast is an exceedingly-rare malignant tumor that shares histological characteristics with osteosarcoma of the bone. Since effective therapies have not yet been established, standard therapy for osteosarcoma of the bone was examined in the present study for efficacy against primary osteosarcoma of the breast in a patient-derived orthotopic xenograft (PDOX) nude-mouse model. Materials and methods The PDOX mouse models were established by surgical implantation of the primary osteosarcoma of the breast specimen into the mammary gland of nude mice. Mice with tumors were randomized into four groups, each n=4: control group; cisplatinum (CDDP)-treatment group; doxorubicin (DOX)-treatment group; and CDDP/DOX-combination-treatment group. Mice were treated for twenty-one days, three weeks after implantation. Tumor size and body weight were measured during three weeks of treatment. Results Significant tumor growth inhibition was observed, compared to the control, in the CDDP-treatment group, the DOX-treatment group, and the combination-treatment-group. Only the combination treatment regressed the tumor. Conclusion CDDP and DOX which are standard first-line therapies for osteosarcoma, may be clinically effective against primary osteosarcoma of the breast, and in particular, their combination.

The goals of this study were to report our clinical experience in a single center at a high-volume tertiary university hospital in Istanbul and to introduce a diagnostic algorithm based on a 5-year follow-up of 152 women with biopsy-proven idiopathic granulomatous mastitis (IGM). IGM is an uncommon, non-malignant, chronic inflammatory disease of the mammary gland with an unknown etiology. The symptoms, clinical presentation, and radiologic findings of IGM may resemble carcinomas. To our knowledge, this study comprises the largest series of IGM, especially with a 5-year follow-up, yet reported in the English literature. The present study reported our single-center clinical experience at a tertiary university hospital in Istanbul, Turkey, and introduced a diagnostic algorithm using our treatment protocol based on a 5-year follow-up of 152 women with biopsy-proven IGM. Our database of 10 years’ experience containing over 700 patients with IGM was evaluated. However, to assess recurrence and resistance to treatment, only the 152 patients with a long follow-up period of 5 years were included in the study group. The analysis included 152 women with biopsy-proven IGM who were treated between January 2009 and March 2014. The clinical data of the presentation, histopathology, and treatment modalities were analyzed by reviewing the medical records. Of the 152 patients diagnosed with granulomatous mastitis, 32 (21%) recovered by expectant management, while 14 (9%) responded to antibiotics, 65 (43%) received corticosteroids, 20 (13%) had antituberculosis medication, 16 (11%) underwent excision, and 5 (3%) responded to non-steroidal anti-inflammatory drugs. Fifty-one patients (33%) had recurrence; of these, 30 achieved a cure with second-line treatment, 16 underwent excision, and 5 achieved a cure with observation. IGM is a rare benign breast disorder, and clinicians need a high index of suspicion to diagnose it, as IGM can be mistaken for breast cancer. Unlike periductal mastitis, IGM does not evolve secondary to nicotine addiction and is typically seen in women of childbearing age with a recent history of pregnancy and lactation. The diagnosis can be challenging, and an evaluation with a multidisciplinary team is necessary. There is no consensus concerning the definitive treatment approach. We suggested a diagnostic algorithm in the present study, using our treatment protocol based on the 5 years’ follow-up of 152 women.

The article deals with a peculiar group of breast diseases, which belongs to the B3 category according to the BI-RADS classification, the so-called lesions with undefined malignant potential. The basic information concerning their biological behavior and modern tactical approaches to the management of patients with similar changes in the mammary gland are presented. The possibility of conservative management, as well as the minimization of surgical aid, in cases of indications for it, up to vacuum aspiration biopsy is being considered. In addition, the significance of these diseases for the subsequent development of pre-invasive and infiltrative cancer is discussed.

Background: Cancer is a life-threatening disease that affects approximately 18 million individuals worldwide. Breast cancer is the most common female neoplasm globally with more than 276,480 new cases of invasive breast cancer expected to be diagnosed in women in the U.S. alone in 2020. Genetic and epigenetic factors play role in the carcinogenesis and progression of this disease. In this study, MCF-7 adenocarcinoma cells were transfected with CRISPR/Cas9 plasmid to either knock out CDK11 or to activate CDH1. Treated cells were allografted into the mammary glands of female rats (150–190 g, 6–8 weeks) to evaluate the capability of these cells to control cancer progression and metastasis. Results: qPCR data revealed a significant downregulation of CDK11 and upregulation of CDH1. Cell cycle analysis and apoptosis assays indicated the knockout of CDK11 and simultaneous activation of CDH1 resulted in cell cycle arrest at G2/M phase and accumulation of cells at G2. Meanwhile, the percentage of cells that underwent late apoptosis increased in both genome editing hits. Histopathological sectioning data indicated that untransfected MCF-7 cells were capable of developing tumors in the mammary gland and initiation g angiogenesis. Transfected cells significantly restricted cancer cell infiltration/invasion by minimally localizing tumors and inhibiting angiogenesis. Conclusions: Although further investigation is needed, the present data indicate the potentiality of using CRISPR/Cas9-based therapy as a promising approach to treat breast cancer. Impact: these data indicate targeting cancer-related genes via any genome editing tool might represent a novel approach to combat cancer. Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabla normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:8.0pt; mso-para-margin-left:0cm; line-height:107%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri",sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;}

Cancer is a major public health problem in all countries and remains as one of the top leading causes of death worldwide. Herein we report the synthesis of novel series of triazoloquinazolines as potential anticancer agents. Structures of this set of compounds were confirmed by different spectral data including IR, 1H NMR, 13C NMR, Mass spectra, and elemental analyses. All the newly synthesized compounds were evaluated for their anti-proliferative activity against four human cancer cell lines namely; Hepatocellular carcinoma (HePG-2), Mammary gland breast cancer (MCF-7), Human prostate cancer (PC3) and Colorectal carcinoma (HCT-116)). Results of cytotoxicity evaluation showed that the synthesized compounds displayed moderate cytotoxic activity against the selected cancer cell lines. Compound 9 showed the highest anti-proliferative effect followed by compound 7 against colorectal carcinoma cell line (HCT-116) with IC50 values of 17.35 and 27.05 µM respectively. As well, both compounds showed moderate activity against hepatocellular carcinoma (HePG2) with IC50 values of 29.47 and 39.41 µM respectively.

We previously showed that dietary trans-10, cis-12 conjugated linoleic acid (10,12 CLA) stimulates estrogen-independent mammary growth in young ovariectomized mice. Here we investigated the effects of in utero or postnatal exposure to cis-9, trans-11 (9,11 CLA) and 10,12 CLA on postnatal development of the mammary gland and its responsiveness to ovarian steroids. In the first experiment we fed dams different CLA prior to and during gestation, then cross fostered female pups onto control fed dams prior to assessing the histomorphology of their mammary glands. Pregnant dams in the second experiment were similarly exposed to CLA, after which their female pups were ovariectomized then treated with 17β-estradiol (E), progesterone (P) or E + P for 5 days. In a third experiment, mature female mice were fed different CLA for 28 days prior to ovariectomy, then treated with E, P or E + P. Our data indicate that 10,12 CLA modifies the responsiveness of the mammary glands to E or E + P when exposure occurs either in utero, or postnatally. These findings underline the sensitivity of the mammary glands to dietary fatty acids and reinforce the potential for maternal nutrition to impact postnatal development of the mammary glands and their risk for developing cancer.

To report the case of a patient diagnosed with ectopic mammary tissue in the vulva, and to conduct a literature review of the diagnosis, treatment and prognosis of this condition in that location.A 49-year-old patient who presented with a painful vulvar mass to a private intermediate complexity center in Bogotá, Colombia. The lesion was assessed on ultrasound and then surgically excised; histopathology showed ectopic mammary tissue with absence of malignancy. A search was conducted in the PubMed, Embase, Cochrane, LILACS and Scielo databases using the keywords “Vulva,” “Breast” and “Ectopic.” Case reports and case series of women with histopathology-confirmed mammary tissue in the vulva were included.Overall, 184 titles were identified and, of these, 94 were ultimately included, for a total of 126 cases, with 57.9% being benign tumors, 95% in women under 50 years of age, and 42.06% being malignant tumors, 92% in women over 50 years of age. Diagnosis was made on the basis of the clinical findings, with ancillary diagnostic imaging, tumor markers and immunohistochemistry in some cases. Local excision was performed in 91% of cases with benign pathology and in 43% of cases with malignant pathology, with the diagnostic method being therapeutic.Ectopic mammary tissue in the vulva must be considered as part of the differential diagnosis of vulvar masses, prognosis being different in pre and postmenopausal women. Further studies are needed to enhance the characterization of this condition and define the ideal course of treatment in terms of relapse and survival.reportar el caso de una paciente con diagnóstico de tejido mamario ectópico en vulva, y realizar una revisión de la literatura acerca del diagnóstico, tratamiento y pronóstico de esta condición en esta localización.paciente de 49 años consulta por masa vulvar dolorosa a un centro privado de nivel medio de complejidad ubicado en Bogotá, Colombia. Se realizó ecografía de la lesión y posteriormente escisión quirúrgica; la histopatología mostró tejido mamario ectópico sin malignidad. Se realizó una búsqueda en las bases de datos PubMed, Embase, Cochrane, LILACS y Scielo, con las palabras clave “Vulva”, “Glándulas mamarias”, “ectópico”, “Vulva”, “Breast” y “Ectopic”, y se incluyeron reportes y series de caso de mujeres con tejido mamario en vulva confirmado por histopatología.se identificaron 184 títulos, de los cuales 94 fueron finalmente incluidos para un total de 126 casos. El 57,9% eran tumores benignos, con un 95% en mujeres menores de 50 años; y el 42,06% eran tumores malignos, el 92 % en mujeres mayores de 50 años. El diagnóstico se realizó por clínica, con imágenes diagnósticas complementarias, marcadores tumorales e inmunohistoquímica en algunos casos. Se realizó escisión local en el 91% de los casos de patología benigna y 43% de patología maligna, siendo el método diagnóstico y terapéutico.el tejido mamario ectópico en vulva debe ser considerado como parte del diagnóstico diferencial de masas a nivel vulvar con diferente pronóstico en mujeres pre y posmenopáusicas. Se requieren más estudios para una mejor caracterización de la patología y definir el tratamiento ideal en términos de recaída y supervivencia.

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.