Search Results (905)

Bovine mastitis is a persistent and challenging illness in dairy industry, bringing about devastating economic losses to the sector. The longstanding over-reliance on antibiotic therapy has raised severe public health concerns, highlighting the critical need to develop safe and effective alternative regimens. Pulsatilla saponin B4 (PSB4), a plant-derived triterpenoid saponin, has been shown to regulate inflammation. However, the effect of PSB4 on mastitis in lactating cows has not been elucidated. The aim of this study was to explore the anti-inflammatory property and the molecular mechanisms of PSB4 in bovine mammary epithelial cells (BMECs) and mouse mastitis model. In cultured cells, PSB4 alleviated LPS-induced inflammatory reaction by suppressing the expression of pro-inflammatory mediators in BMECs. Notably, RNA-seq analysis indicated that the anti-inflammatory effect of PSB4 was correlated with IL-17RA and NF-κB signaling, with subsequent Western blot validation. Moreover, BMECs were stimulated with recombinant interleukin-17A (rIL-17A) to induce inflammation and challenged with Taltz to specifically inhibit IL-17RA signaling. The results showed that PSB4 reversed the rIL-17A-induced upregulation of elements within the IL-17RA pathway and its downstream MAPK/NF-κB cascade, including their downstream effectors. Furthermore, Taltz blocked the efficacy of PSB4 in protecting against LPS-induced inflammation. In vivo, PSB4 alleviated the inflammatory damage of mammary gland, pro-inflammatory mediator levels in mammary gland tissue and se-rum, while blocking the activation of IL-17RA signal along with downstream MAPK/NF-κB signal in LPS-induced mouse mastitis model. Taken together, studies in both BMECs and mice demonstrated that PSB4 alleviates mastitis by inhibiting IL-17RA signaling and downstream MAPK/NF-κB pathway, which may be a new strategy and a target for the management of mastitis. Full article

Bovine mastitis remains a major impediment to optimal dairy production. Somatic cell count (SCC) is commonly used as an indicator of mammary gland inflammation, while milk microbiota may also reflect mastitis-related changes. Here, we employed Oxford Nanopore full-length transcript sequencing to delineate the peripheral blood transcriptomic landscape of Xinjiang Brown cattle stratified by high (SCC ≥ 1,000,000 cells mL⁻¹) and low (SCC ≤ 200,000 cells mL⁻¹) SCCs, with the objective of identifying candidate genes underpinning mastitis resistance. We identified 226 differentially expressed genes and 441 differentially expressed transcripts. Genes in the high-SCC group were prominently enriched in immune response pathways and chemokine signalling cascades. Protein–protein interaction network analysis further delineated a core module of ten immune-related genes, including CCL4, IL1B and CXCL2. Integrative analysis with complementary second-generation sequencing data pinpointed CXCL2 as a high-priority candidate. Subsequent RT–qPCR and enzyme-linked immunosorbent assay (ELISA) validation revealed that CXCL2 expression was significantly elevated both in high-SCC individuals and in an LPS-induced bovine mammary epithelial cell inflammation model. Collectively, these findings establish CXCL2 as a putative molecular marker for mastitis resistance breeding and provide a foundational resource for deciphering the molecular mechanisms governing mammary health. Full article

Although human mammary glands were traditionally considered sterile, accumulating evidence has established the presence of distinct microbial communities that may have colonized breast tissue primarily via retrograde nipple flow or via hematogenous or lymphatic translocation from other body sites. Comparative studies reveal differences in the microbiota of healthy and diseased breast tissues, with variations in microbial signatures across breast cancer subtypes and in comparison with adjacent normal tissues. This review synthesizes current evidence on the composition of the breast microbiome, the factors shaping its development, and alterations it undergoes in inflammatory and malignant breast diseases. Furthermore, the article discusses mechanistic insights, methodological challenges, and future therapeutic perspectives based on published studies employing culture-independent approaches, such as 16S rRNA gene sequencing and metagenomic analyses. Key host-related factors influencing breast-associated microbial communities, including hormonal regulation, environmental exposure, diet, and therapeutic interventions, are explored. The existing literature is assessed to identify key associations between the breast microbiome and host signaling pathways, as well as the significant challenges that remain unresolved, including low biomass contamination, inter-study variability, limited longitudinal data, and an incomplete understanding of causality. Addressing these limitations is critical for advancing microbiome-based diagnostic and therapeutic strategies for breast disease. Full article

Heat stress (HS) is among the most economically consequential environmental challenges to global dairy production, causing progressive declines in milk yield, compositional quality, and mammary cellular integrity. The temperature–humidity index (THI) is the primary thermal load metric, with performance-impairment thresholds typically beginning at THI 68 in Holstein cattle, with severe impacts manifesting beyond THI 72; breed-specific thresholds for Jersey, Brown Swiss, and Simmental cows differ owing to their lower metabolic heat load and greater inherent thermotolerance. At the molecular level, HS activates heat shock protein networks—notably HSPA1A, HSP90B1, and HSPH1—through HSF1/HSF4 transcriptional activation, while simultaneously suppressing casein genes (CSN1S1, CSN2, CSN3), lipogenic genes (FASN, SCD, CD36), amino acid transporters (SLC7A5, SLC38A2), and mTOR-AKT-STAT5 translational machinery, collectively impairing milk biosynthetic capacity. Pro-apoptotic signaling (BAX, CASP3 upregulation; BCL2 downregulation) and mitochondrial dysfunction further compromise mammary epithelial viability. Post-transcriptional regulation through miRNA, circRNA, and lncRNA competing endogenous RNA networks, alongside epitranscriptomic m6A modifications, adds further regulatory complexity. Genome-wide association studies have identified SNPs in HSP70A1A, HSPA4, TLR4, and PRLR as thermotolerance candidates compatible with sustained milk production. Nutritional supplementation with methionine, arginine, and taurine partially restores cellular synthetic capacity. Integrating multi-trait genomic selection with Bos indicus introgression, precision cooling, and targeted nutrition offers the most viable path toward climate-resilient, high-producing dairy cattle. Full article

Pleiotrophin (PTN), a heparin-binding growth factor with potent mitogenic and angiogenic activity, has emerged as a key regulator of mammary gland biology and a potential driver of breast cancer progression. This review integrates current evidence on PTN’s roles from normal mammary development, where it can delay ductal outgrowth, to triple negative breast cancer, where it promotes lung metastasis and correlates with poor survival. Though frequently reported as being overexpressed in breast cancer, the published data indicates that PTN transcription is reduced in cancer relative to normal breast cells. By contrast, serum PTN protein levels have been shown by multiple studies to be elevated in breast cancer patients relative to healthy controls. We examine the expression and function of PTN at a cellular level and explore the interplay between PTN and the tumour microenvironment. We evaluate preclinical models, clinical correlations, and emerging biomarker data that position PTN as a candidate prognostic indicator and therapeutic target. Despite growing interest, significant gaps remain regarding context-specific signalling. By integrating developmental and oncogenic perspectives, this review highlights PTN as a pivotal but underexplored factor in mammary gland physiology and breast cancer and outlines future research directions needed to translate PTN-targeted strategies into clinical benefit. Full article

Under grazing conditions, it is difficult for lactating Yili mares to meet their nutritional requirements and those of their suckling foals solely through the consumption of natural pasture. Furthermore, seasonal variations and rainfall significantly influence the quality and nutrient content of forage, which severely constrains the healthy breeding of Yili horses and the industrial development of mare milk resources. Therefore, this study aimed to investigate the effects of concentrate supplementation on lactation performance and milk concentrations of amino acids, fatty acids, and mineral elements in Yili horses under grazing conditions. Twenty-two healthy Yili mares in early lactation, with similar ages (3–4 years), foaling dates, and body weights (391.5 ± 13.74 kg), were randomly assigned to either a grazing group (G, n = 11) or a grazing + supplementation group (GS, n = 11). Mares in group G grazed naturally on pasture, while those in group GS received 1 kg of concentrate supplement twice daily (totaling 2 kg/day) in addition to grazing. The experimental period lasted for 100 days, including a 10-day adaptation period and a 90-day formal experimental period. The results showed that: (1) In terms of lactation performance, the GS group exhibited highly significant increases in milk yield and lactose yield (p < 0.01), as well as significant increases in milk protein and milk fat yields (p < 0.05), with an extended duration of the peak lactation period. (2) Regarding the amino acid profile, the concentrations of threonine (Thr), serine (Ser), glycine (Gly), and alanine (Ala) in the milk of the GS group were significantly higher than those in the G group (p < 0.05), whereas the proline (Pro) content was significantly lower (p < 0.01); supplementation improved the uptake of certain functional amino acids by the mammary gland. (3) Concerning the fatty acid profile, the concentrations of polyunsaturated fatty acids (PUFA) and alpha-linolenic acid in the milk of the G group were significantly or highly significantly higher than those in the GS group (p < 0.05 or p < 0.01). (4) For mineral elements, concentrate supplementation highly significantly decreased the potassium (K) content and the K/Na ratio in horse milk (p < 0.01), highly significantly increased the levels of iron (Fe) and cobalt (Co) (p < 0.01), and significantly enhanced the chromium (Cr) content (p < 0.05). In conclusion, concentrate supplementation during grazing improved lactation performance in Yili mares, primarily by increasing milk yield and extending the peak lactation period. However, grazing alone was more favorable for maintaining higher PUFA and α-linolenic acid proportions in milk. Therefore, concentrate supplementation should be regarded as a nutritional strategy that increases milk output and modifies amino acid and mineral element composition, but may involve a trade-off with some beneficial fatty acids. Full article

Cadmium (Cd) is a ubiquitous heavy metal environmental toxicant, and exposure poses a persistent public health concern due to its bioaccumulative properties. However, the impact of maternal Cd exposure on mammary gland function during lactation remains insufficiently understood. In this study, we investigated the effects of Cd exposure during pregnancy and lactation on mammary gland development, hormonal homeostasis, and lactational performance using a drinking water exposure model. Pregnant 7-week-old ICR mice were randomly assigned into Control and Cd groups. The Control group received standard drinking water, while the Cd group was administered water supplemented with Cd (12 mg/L). Results demonstrated that maternal Cd exposure markedly reduced maternal and offspring weight, respectively. Histological analysis revealed that Cd exposure resulted in decreased numbers of mammary alveoli and widened interstitial spaces, indicating impaired mammary development. In addition, Cd exposure markedly compromised lactational performance, as reflected by reduced milk yield and decreased levels of milk protein, fat, and lactose across multiple lactation stages. Integrated metabolomic and transcriptomic analyses further revealed substantial alterations in hormonal profiles and metabolic pathways associated with mammary gland function. Notably, Cd accumulation was detected in mammary tissue and milk, accompanied by transcriptional changes in genes involved in milk synthesis and lipid metabolism. In conclusion, these findings indicated that maternal Cd exposure during pregnancy and lactation is associated with structural and functional alterations of the mammary gland, leading to reduced lactational performance and impaired offspring growth. The findings contribute to a better understanding of the risks posed by environmental Cd exposure, emphasizing the need for effective public health strategies to mitigate its impact on maternal and infant health. Full article

Introduction: The mammary gland is highly sensitive to ionising radiation, making the average glandular dose (AGD) the most appropriate metric for breast dosimetry. With increasing participation in breast screening programmes, cumulative radiation exposure remains a concern. This study aimed to investigate the correlation between AGD and exposure parameters under controlled conditions of compressed breast thicknesses and mammographic projections. Methods: A retrospective, cross-sectional analysis evaluated data from 609 patients who underwent clinically justified digital mammography examinations with a single direct digital radiography system at King Abdulaziz Medical City, Jeddah, Saudi Arabia, between September 2023 and September 2025. The Shapiro–Wilk normality test indicated that AGD data were not normally distributed. Consequently, the Wilcoxon signed-rank test was used to assess differences between craniocaudal (CC) and mediolateral oblique (MLO) projections for the right and left breasts across CBT ranges. Spearman’s rank correlation coefficient was used to evaluate associations between AGD and exposure parameters (compression force (CF), mAs, and kVp) under controlled conditions of CBT ranges and projections. Results: AGD increased with CBT, rising from 1.36–1.39 mGy at 30–39 mm to 2.25–3.05 mGy at 70–79 mm. MLO projections consistently showed higher AGD than CC projections, with greater differences from 50–59 mm. Statistically significant differences were observed in projections across CBT. Spearman’s analysis demonstrated a significant positive correlation between AGD and CBT (p < 0.0001), which was strongest in the right MLO (ρ = 0.5082). Within the 50–59 mm range, AGD strongly correlated with mAs and moderately with kVp, but not with CF. Conclusions: AGD increases significantly with CBT, particularly in MLO projections. Within the 50–59 mm CBT range, mAs showed the strongest influence on AGD, while kVp had a moderate effect and CF was non-significant. These findings support dose optimisation through exposure parameter control and alignment with DRLs while maintaining image quality. Full article

The river buffalo is an economically important livestock species supplying milk and meat. However, a multi-tissue transcriptomic atlas for the key dairy river buffalo breeds, Murrah and Nili-Ravi, has not yet been established, and the lack of stable reference genes has hindered in-depth studies of their biological functions and the molecular mechanisms underlying key economic traits such as lactation. We established a multi-tissue gene expression atlas across 20 tissues and identified 717 housekeeping genes (HKGs), and RPL37A and EEF2 were further shown to be stable candidate reference genes under the conditions tested. We found 8368 tissue-specific genes (TSGs), predominantly enriched in the reproductive system. Exploratory analysis of mammary tissue (dry-period vs. public lactating samples, confounded by batch effects) revealed mammary-enriched hub genes including LALBA; these findings are preliminary and require validation. Dynamic analysis across lactation stages (early, peak, mid-, late) identified candidate genes including SEC14L2 and ACSM3. Phenotypic data showed strong negative correlations between milk yield and protein/fat content, and a positive correlation with lactose content. However, causal or regulatory roles were not inferred due to lack of paired individual-level data. Cross-dataset comparisons are descriptive only, and are not key conclusions. In summary, this study lays the foundation for advancing research in lactation trait genetics and functional genomics in river buffalo, with novel reference genes and lactation stage-specific transcriptional dynamics as its main contributions. Full article

Obesity is recognized as a risk factor for breast cancer development and progression. Adipocytes exert their oncogenic effects through complex and interconnected biological mechanisms that encompass metabolic dysfunction, chronic low-grade inflammation, and systemic endocrine alterations. Herein, we reviewed the current evidence explaining how obesity induces a state that reprograms adipose tissue and remodels the breast cancer tumor microenvironment (TME). We first discuss the systemic and local mechanisms linking obesity to inflammation and how these alterations reshape the functional organization of the mammary gland. Then, we discuss how the chronic exposure to tumor-derived signals, together with the altered metabolic state of obese adipose tissue, induces a functional reprogramming of adipocytes, giving rise to so-called cancer-associated adipocytes (CAAs), which actively contribute to tumor progression. Also, the strengths and limitations of biological models to study the crosstalk between adipocytes and tumor cells, including two-dimensional (2D) monolayers and three-dimensional (3D) cell cultures, as well as animal models, are discussed. Special emphasis is placed on 3D co-culture models, which more accurately reproduce spatial organization, direct cell–cell contact, and diffusion dynamics, providing a more physiologically relevant environment for studying how obesity and inflammation reshape the TME in breast cancer. Finally, we highlight the limitations of conventional experimental models and review recent advances in 3D-based platforms, emphasizing their mechanistic insights and translational potential. Full article

2,3′,4,4′,5-Pentachlorobiphenyl (PCB118) is a persistent environmental pollutant associated with adverse female reproductive outcomes; however, its effects on uterine function and epigenetic regulation remain incompletely understood. This study investigated whether PCB118 disrupts uterine decidualization and angiogenesis through miRNA-mediated regulatory pathways. Human endometrial stromal cells (HESCs) and human umbilical vein endothelial cells (HUVECs) were exposed to an environmentally relevant, non-cytotoxic concentration of PCB118. Decidualization and angiogenesis were evaluated in vitro, and underlying mechanisms were investigated using molecular and miRNA-based approaches. In vivo validation of miR-542-3p expression was performed in pregnant mice following PCB118 exposure. PCB118 exposure was associated with reduced expression of decidualization markers, including prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP-1), as well as impaired angiogenic capacity in HUVECs. PCB118 treatment was accompanied by increased miR-542-3p expression, which was associated with decreased integrin-linked kinase (ILK) levels and changes in transforming growth factor beta 1 (TGF-β1) and total Smad2 protein abundance. ILK overexpression partially restored decidualization and angiogenesis-related phenotypes, supporting a functional involvement of ILK in these processes. Consistently, elevated miR-542-3p expression was observed in murine endometrial tissues following PCB118 exposure, suggesting physiological relevance in vivo. PCB118 exposure is associated with impaired decidualization and angiogenesis, potentially involving dysregulation of the miR-542-3p/ILK signaling axis, suggesting a potential role for epigenetic modulation in PCB118-associated reproductive dysfunction. Full article

Mastitis is a common inflammatory disease that harms mammary gland health. Its development is closely linked to dysregulated inflammatory signaling. Eicosapentaenoic acid (EPA), an omega-3 polyunsaturated fatty acid, has potential anti-inflammatory effects. However, its molecular mechanism in mastitis prevention remains unclear. In this study, we used both in vivo and in vitro models to evaluate how EPA pretreatment regulates mastitis-related inflammatory signaling. Transcriptome analysis showed that differentially expressed genes after EPA treatment were mainly enriched in the peroxisome proliferator-activated receptor (PPAR) signaling pathway. In an LPS-induced mastitis model, EPA restored the LPS-reduced PPARγ protein level and suppressed NF-κB p65 activation, consistent with reduced nuclear translocation of p65. Similar effects were observed in mammary epithelial cells, where EPA inhibited NF-κB activation at 50 and 100 μM. Functional experiments further showed that a PPARγ agonist mimicked the inhibitory effect of EPA on p65, whereas PPARγ antagonist partially abrogated EPA-mediated inhibition of p65. Collectively, these data indicate that EPA attenuates mastitis-associated inflammation at least in part through the PPARγ–NF-κB axis. Full article

Milk fat synthesis in dairy cows is a complex process affected by ruminal fermentation, systemic metabolism, and mammary gland activity. To explore the metabolic interplay across these systems, a multi-tissue metabolomics approach (rumen fluid, plasma, and milk) using ultra-high-performance liquid chromatography–mass spectrometry was used to identify metabolic differences between Chinese Holstein cows with high (H-MF, 5.82 ± 0.41%) and low (L-MF, 3.60 ± 0.12%) milk fat content under the same diet. The bovine mammary epithelial cells (BMECs) were also cultured to evaluate the impact of a key metabolite, malic acid (MA), on lipid metabolism. Our findings reveal distinct metabolic profiles across rumen fluid, plasma, and milk, with 96, 109, and 79 differential metabolites, respectively, between the L-MF and H-MF groups. In rumen fluid, H-MF cows showed higher levels of lauric acid and succinic acid, linked to fatty acid biosynthesis, while the L-MF cows had elevated citraconic and orotic acids, associated with amino acid metabolism and liver stress. Plasma from the H-MF cows contained higher β-hydroxybutyric acid, methionine sulfoxide, and phosphatidylcholine, supporting lipogenesis, whereas L-MF plasma showed increased 3-hydroxy-L-proline, indicating tissue catabolism. In milk, the L-MF cows had higher MA, while the H-MF cows exhibited elevated L-carnitine, linked to fatty acid β-oxidation. Metabolite trend analysis during rumen fluid–plasma–milk showed that 211 metabolites were classified into 8 profiles. Profile 1 had the largest number of metabolites whose levels were down-regulated from rumen to plasma and enriched in lipid metabolism. Profile 3 (mainly related to amino acid metabolism) and profile 4 (mainly related to energy metabolism) exhibited opposite trends from plasma to milk. In vitro, 200 μM of MA reduced the triglyceride content in BMECs and down-regulated lipogenic genes and their protein expression levels (fatty acid synthase, stearoyl-CoA desaturase and sterol regulatory element binding protein 1). These results highlight how rumen fluid, plasma, and milk metabolites collectively influence milk fat synthesis, with MA acting as a key regulator of lipid metabolism in mammary epithelial cells. Full article

Bovine mastitis is a multifactorial inflammatory disease primarily characterized by inflammatory cell infiltration and the destruction of mammary alveoli. It is a major cause of reduced milk yield and quality. The imbalance between antioxidant defenses and the generation of reactive oxygen species (ROS), which occurs due to the high metabolic activity of the mammary gland during the periparturient period, increases the incidence of mastitis. During early lactation, especially in high-yielding dairy cows, the massive synthesis and secretion of milk increase the energy demand of mammary tissue, leading to excessive ROS accumulation. This results in cell membrane disruption and, ultimately, antioxidant dysfunction in the mammary tissue. This study established an in vitro oxidative stress model by treating bovine mammary epithelial cells (BMECs) with 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH). The optimal concentration of 1000 μmol/L AAPH was determined using the CCK-8 assay. Model validation showed that, compared to the control group, ROS levels were significantly elevated (p < 0.001) and mitochondrial membrane potential was significantly decreased (p < 0.001) in the AAPH-treated group. Transmission electron microscopy (TEM) analysis revealed that AAPH treatment caused ultrastructural damage, including reduced microvilli, mitochondrial swelling, disappearance of cristae, and vacuolization. Mechanistic studies demonstrated that AAPH treatment significantly upregulated the mRNA and protein expression of AMPK, HMOX-1, mTOR, NOS, and SOD (p < 0.001), while significantly downregulating CYP1A1 expression (p < 0.001). Pretreatment with N-acetylcysteine (NAC) effectively alleviated the oxidative stress damage caused by AAPH. This study successfully established an in vitro AAPH-induced oxidative stress model in BMECs and revealed its molecular mechanism of cellular damage. The damage occurs through modulation of the AMPK/mTOR signaling pathway and the regulation of antioxidant-related gene expression. Full article

Background: Breast cancer remains a major public health problem, with increasing incidence and persistent survival disparities. In Romania, barriers to early diagnosis and access to multidisciplinary treatment may contribute to poorer outcomes. Methods: We conducted a retrospective single-center cohort study including 118 women diagnosed with and/or treated for breast cancer in our institution between 1 January and 31 December 2020. Patients were followed for 5 years. The primary outcome was overall survival (OS). Clinicopathological characteristics, treatment exposure, pathological response after neoadjuvant therapy, and factors associated with OS were analyzed. Results: The median age at diagnosis was 62 years. Most tumors were located in the upper quadrants, and the most frequent subtype was hormone receptor-positive/HER2-negative breast carcinoma. During follow-up, 26.27% of patients died from disease progression or associated complications. Estimated OS was 88.7% at 1 year and 72.8% at 5 years. Older age at diagnosis and treatment exposure patterns, including the absence of neoadjuvant therapy, were associated with OS. Conclusions: In this single-center retrospective cohort, overall survival was associated with age at diagnosis, tumor characteristics, and treatment patterns. The high proportion of early deaths and the frequent absence of documented surgical treatment in patients who died suggest important challenges related to late presentation, continuity of care, and access to guideline-concordant multidisciplinary treatment in the Romanian setting. Full article