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Keywords = mammary gland health

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22 pages, 2181 KiB  
Review
Incorporating Morphological Evaluations into Breeding Soundness Examinations for Female Dogs
by Dane Wells Schwartz, Jonah Kvernum, Naomie Macias, Muhammed Salman Waqas and Michela Ciccarelli
Animals 2025, 15(14), 2045; https://doi.org/10.3390/ani15142045 - 11 Jul 2025
Viewed by 687
Abstract
This article highlights the importance of evaluating the morphological characteristics of female dogs during breeding soundness examinations (BSEs) to assess their reproductive potential and ensure the health of future offspring. Key traits considered in this evaluation include body type, body condition score, skull [...] Read more.
This article highlights the importance of evaluating the morphological characteristics of female dogs during breeding soundness examinations (BSEs) to assess their reproductive potential and ensure the health of future offspring. Key traits considered in this evaluation include body type, body condition score, skull size and shape, mammary gland conformation, and coat quality. Each of these factors plays a significant role in the health and fertility of breeding females. For example, deviations from breed standards in body size can lead to complications during pregnancy and whelping, while an inappropriate body condition score may disrupt hormonal balance and reproductive cycles. This review also addresses concerns related to brachycephalic breeds, whose conformation can contribute to respiratory and reproductive issues. Additionally, assessing vertebral and pelvic conformation is crucial to prevent dystocia and other complications during delivery. By systematically evaluating these morphological traits, veterinarians can promote ethical breeding practices that prioritize the welfare and genetic health of both breeding females and their puppies. Therefore, integrating comprehensive morphological evaluations into BSEs is essential for responsible dog breeding management, ultimately supporting better reproductive outcomes and healthier future generations. Full article
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23 pages, 3759 KiB  
Review
Highly Pathogenic Avian Influenza (H5N1) Clade 2.3.4.4b in Cattle: A Rising One Health Concern
by Ivan Camilo Sanchez-Rojas, D. Katterine Bonilla-Aldana, Catherin Lorena Solarte-Jimenez, Jorge Luis Bonilla-Aldana, Jaime David Acosta-España and Alfonso J. Rodriguez-Morales
Animals 2025, 15(13), 1963; https://doi.org/10.3390/ani15131963 - 3 Jul 2025
Viewed by 1036
Abstract
Highly pathogenic avian influenza (HPAI) H5N1, particularly clade 2.3.4.4b, has demonstrated an unprecedented capacity for cross-species transmission, with recent reports confirming its presence in dairy cattle in the United States of America (USA) in 2024. This unexpected spillover challenges traditional understanding of the [...] Read more.
Highly pathogenic avian influenza (HPAI) H5N1, particularly clade 2.3.4.4b, has demonstrated an unprecedented capacity for cross-species transmission, with recent reports confirming its presence in dairy cattle in the United States of America (USA) in 2024. This unexpected spillover challenges traditional understanding of the virus’s host range and raises serious public health and veterinary concerns. Infected cattle presented with clinical signs such as decreased milk production, thickened or discolored milk, respiratory issues, and lethargy. Pathological findings revealed inflammation of the mammary glands and the detection of a virus in nasal secretions and raw milk, suggesting a potential for both intra- and interspecies transmission. While the current risk of human-to-human transmission remains low, the detection of H5N1 in a human exposed to infected cattle highlights the need for heightened surveillance and protective measures. Moreover, the presence of infectious viruses in the food chain, particularly in unpasteurized milk, introduces a new dimension of zoonotic risk. This review synthesizes emerging evidence on the epidemiology, pathology, diagnostic findings, and zoonotic implications of HPAI H5N1 infection in cattle. It also highlights the importance of genomic surveillance, intersectoral collaboration, and One Health approaches in managing this evolving threat. As the virus continues to circulate and adapt across diverse hosts, including wild birds, domestic poultry, and now mammals, the potential for reassortment and emergence of novel strains remains a significant concern. Immediate actions to strengthen biosecurity, monitor viral evolution, and protect both animal and human populations are critical to mitigate the global risk posed by this expanding panzootic. Full article
(This article belongs to the Special Issue Infection Immunity, Diagnosis and Prevention of Avian Influenza)
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12 pages, 4848 KiB  
Brief Report
Clinical Mastitis in Small Ruminants Referred to a Veterinary Teaching Hospital: 23 Cases
by Gabriel Inácio Brito, Liz de Albuquerque Cerqueira, Simone Perecmanis, José Renato Junqueira Borges, Márcio Botelho de Castro and Antonio Carlos Lopes Câmara
Microorganisms 2025, 13(7), 1512; https://doi.org/10.3390/microorganisms13071512 - 28 Jun 2025
Viewed by 1180
Abstract
Clinical mastitis in small ruminants is usually seen with an incidence of less than 5% and most cases, especially with hyperacute evolution, are not referred for hospital care. During the 5-year survey, 16 goats and 7 sheep, totaling 23 small ruminants, met the [...] Read more.
Clinical mastitis in small ruminants is usually seen with an incidence of less than 5% and most cases, especially with hyperacute evolution, are not referred for hospital care. During the 5-year survey, 16 goats and 7 sheep, totaling 23 small ruminants, met the inclusion criteria with a definitive diagnosis of clinical mastitis. Clinical signs ranged greatly among cases, varying from septic state in hyperacute cases, and enlarged, pendulous udder associated with chronic pain and abnormal gait in chronic cases. Microbiological culture revealed a wide array of bacterial pathogens, including Staphylococcus aureus, Escherichia coli, Streptococcus spp., and Pasteurella spp. In vitro antimicrobial susceptibility profiles varied greatly among bacteria isolates, ranging from sensitive to all tested antimicrobials to a multi-resistant profile. Pathological features included hyperemia and dark red areas of necrosis in the skin, marked hyperemia of the affected gland at the cut surface, lactiferous ducts and gland cisterns filled by cloudy or suppurative fluid, abscesses, and hardness of the mammary gland parenchyma. This retrospective study highlights the multifactorial nature and clinical variability of mastitis in small ruminants, demonstrating its significant impact on animal health, welfare, and production. Full article
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12 pages, 1029 KiB  
Article
Structural and Hormonal Changes in Reproductive-Age Women Post-COVID-19: A Cross-Sectional Ultrasound and Biochemical Study
by Sandugash Yerkenova, Vyacheslav Lokshin, Almagul Kurmanova, Sharapat Moiynbayeva, Galiya Alikeyeva, Gulnara Kalibekova, Tamara Abdirova, Zamira Zhantenova, Kuralay Shaikova, Alla Mireeva and Aknur Turgumbayeva
Diagnostics 2025, 15(12), 1536; https://doi.org/10.3390/diagnostics15121536 - 17 Jun 2025
Viewed by 633
Abstract
Background/Objectives: The long-term impact of COVID-19 on female reproductive health remains poorly understood. This study aimed to assess structural and endocrine alterations in women of reproductive age who had recovered from SARS-CoV-2 infection compared to uninfected controls. Materials and Methods: A total of [...] Read more.
Background/Objectives: The long-term impact of COVID-19 on female reproductive health remains poorly understood. This study aimed to assess structural and endocrine alterations in women of reproductive age who had recovered from SARS-CoV-2 infection compared to uninfected controls. Materials and Methods: A total of 150 women aged 18–45 years were enrolled in a comparative study: 75 with a confirmed history of COVID-19 and 75 without one. All participants underwent ultrasound examinations of their pelvic organs and mammary glands, along with laboratory assessment of reproductive hormones and inflammatory markers. Results: Structural abnormalities in the pelvic organs were observed in 53.5% of the post-COVID-19 group versus 12.0% of the control group (p < 0.001), with oophoritis showing a statistically significant association (OR = 11.38; 95% CI: 1.42–91.36; p = 0.009). Non-significant but elevated frequencies were also found for uterine fibroids and breast cysts. Biochemically, post-COVID-19 participants demonstrated higher serum ferritin, estradiol, and fibrinogen levels, along with lower TSH and AMH levels, suggesting potential endocrine disruption and persistent inflammation. Conclusions: Women with a history of COVID-19 may be at increased risk of developing structural and hormonal abnormalities, highlighting the importance of post-infection gynecological and endocrine monitoring. Further longitudinal studies are required to elucidate the long-term effects and underlying mechanisms of these alterations. Full article
(This article belongs to the Special Issue Biochemical Testing Applications in Clinical Diagnosis)
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28 pages, 1191 KiB  
Review
piRNAs as Potential Regulators of Mammary Gland Development and Pathology in Livestock
by Wenjing Yu, Zixuan Zhang, Zhonghua Wang, Xusheng Dong and Qiuling Hou
Vet. Sci. 2025, 12(6), 594; https://doi.org/10.3390/vetsci12060594 - 17 Jun 2025
Viewed by 699
Abstract
PiRNAs are a subclass of non-coding RNAs, 26–31 nucleotides (nt) in length, that form regulatory complexes through their interaction with PIWI proteins. Studies in model organisms have demonstrated that piRNAs play crucial roles in tissue development and in predicting disease outcomes, positioning them [...] Read more.
PiRNAs are a subclass of non-coding RNAs, 26–31 nucleotides (nt) in length, that form regulatory complexes through their interaction with PIWI proteins. Studies in model organisms have demonstrated that piRNAs play crucial roles in tissue development and in predicting disease outcomes, positioning them as promising targets for developmental regulation and therapeutic intervention. In contrast, research on piRNAs in animal husbandry is still in its early stages and has not received sufficient attention. Despite this, the few studies available in livestock research have revealed that piRNAs serve as key regulators of reproductive development, underscoring their significant regulatory potential in farm animals and justifying further investigation. Accordingly, this review uses the bovine mammary gland as an exemplary case to summarize the progress in piRNA research related to mammary development and disease. The role of piRNAs in regulating breast cancer stem cell proliferation and modulating inflammatory progression is a highly active area of research. We hypothesize that piRNAs may play a potential role in regulating both mammary gland development and mastitis, making them promising targets for enhancing mammary development and overall health in dairy cattle and providing a theoretical foundation for further piRNA applications in animal husbandry. Full article
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5 pages, 176 KiB  
Commentary
Highly Pathogenic Avian Influenza A(H5N1) Virus: How Far Are We from a New Pandemic?
by Giovanni Di Guardo
Vet. Sci. 2025, 12(6), 566; https://doi.org/10.3390/vetsci12060566 - 9 Jun 2025
Cited by 2 | Viewed by 1128
Abstract
The focus of this commentary is represented by the pandemic risk associated with the highly pathogenic avian influenza (HPAI) A(H5N1) virus, clade 2.3.4.4b. More in detail, the herein dealt pandemic alarm appears to be primarily justified by the huge and progressively growing number [...] Read more.
The focus of this commentary is represented by the pandemic risk associated with the highly pathogenic avian influenza (HPAI) A(H5N1) virus, clade 2.3.4.4b. More in detail, the herein dealt pandemic alarm appears to be primarily justified by the huge and progressively growing number of virus-susceptible domestic and wild birds and mammals, including threatened marine mammal species like South American sea lions and elephant seals as well as harbour porpoises, bottlenose dolphins and polar bears. Of major concern is the susceptibility of dairy cattle to HPAI A(H5N1) virus, particularly the documented and unprecedented colonization of host’s mammary gland tissue, resulting in viral shedding through the milk alongside a large series of cases of infection in dairy farm workers in several USA locations. Despite well-documented zoonotic capability, no evidences of a sustained and efficient HPAI A(H5N1) viral transmission between people have been hitherto reported. If this were to happen sooner or later, a new pandemic might consequently arise. Therefore, keeping all this in mind and based upon the lessons taught by the COVID-19 pandemic, a “One Health, One Earth, One Ocean”-centered approach would be absolutely needed in order to deal in the most appropriate way with the HPAI A(H5N1) virus-associated zoonotic and pandemic risk. Full article
(This article belongs to the Section Veterinary Microbiology, Parasitology and Immunology)
12 pages, 1832 KiB  
Article
Single-Cell Analysis of Host Responses in Bovine Milk Somatic Cells (bMSCs) Following HPAIV Bovine H5N1 Influenza Exposure
by Gagandeep Singh, Sujan Kafle, Patricia Assato, Mankanwal Goraya, Igor Morozov and Juergen A. Richt
Viruses 2025, 17(6), 811; https://doi.org/10.3390/v17060811 - 3 Jun 2025
Viewed by 780
Abstract
The 2024 outbreak of highly pathogenic avian influenza virus (HPAIV) H5N1 in U.S. dairy cattle presented an unprecedented scenario where the virus infected bovine mammary glands and was detected in milk, raising serious concerns for public health and the dairy industry. Unlike previously [...] Read more.
The 2024 outbreak of highly pathogenic avian influenza virus (HPAIV) H5N1 in U.S. dairy cattle presented an unprecedented scenario where the virus infected bovine mammary glands and was detected in milk, raising serious concerns for public health and the dairy industry. Unlike previously described subclinical influenza A virus (IAV) infections in cattle, H5N1 infection induced severe clinical symptoms, including respiratory distress, mastitis, and abnormal milk production. To understand the host immune responses and changes, particularly in the mammary gland, we performed single-cell RNA sequencing analysis on bovine milk somatic cells (bMSCs) in vitro exposed to an H5N1 isolate from an infected dairy farm. We identified ten distinct cell clusters and observed a shift toward type-2 immune responses, characterized by T cells expressing IL13 and GATA3, and three different subtypes of epithelial cells based on the expression of genes associated with milk production. Our study revealed temporal dynamics in cytokine expression, with a rapid decline in luminal epithelial cells and an increase in macrophages and dendritic cells, suggesting a role in increased antigen presentation. While viral RNA was detected in bulk-exposed bMSC samples via qRT-PCR, no viral reads were observed in the scRNA-seq data, indicating that the immune responses captured may be due to exposure to viral components rather than productive infection. This research fills a critical gap in understanding the immune responses of bovine mammary glands to H5N1 exposure and highlights the need for further investigation into therapeutic strategies for managing such outbreaks. Full article
(This article belongs to the Special Issue Advances in Endemic and Emerging Viral Diseases in Livestock)
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17 pages, 1412 KiB  
Article
The Relationship Between Protein Fraction Contents and Immune Cells in Milk
by Haitong Wang, Xiaoli Ren, Li Liu, Zhuo Yang, Chunfang Li, Xiangnan Bao, Ayihumaer Amantuer, Peipei Wen, Dongwei Wang and Shujun Zhang
Animals 2025, 15(11), 1578; https://doi.org/10.3390/ani15111578 - 28 May 2025
Viewed by 364
Abstract
Mastitis significantly impacts both the yield and quality of milk. The somatic cell count (SCC) and differential somatic cell count (DSCC), which are related to immune cells, are primary indicators for assessing mammary gland health. In this study, eight previously established mid-infrared spectroscopy [...] Read more.
Mastitis significantly impacts both the yield and quality of milk. The somatic cell count (SCC) and differential somatic cell count (DSCC), which are related to immune cells, are primary indicators for assessing mammary gland health. In this study, eight previously established mid-infrared spectroscopy models were utilized to predict the content of milk protein fractions (αs1-CN, β-CN, κ-CN, total CN, α-LA, β-LG, IgG, and LF) in milk samples from 21,388 lactating cows across 33 herds. Four linear mixed models were applied to analyze the secretion patterns of milk protein fractions by days in milk (DIM) and parity, their variations under different mastitis conditions, and their associations with the somatic cell score (SCS), DSCC, and immune cell counts (PMN + LYM score (PMN + LYMS) and MAC score (MACS)). The primary findings of the investigation comprised the following: (1) IgG was higher in early lactation, decreased with advancing lactation days, and slightly increased in late lactation, while seven other protein factions decreased from early to peak lactation and increased during mid-to-late lactation. Parity influenced all milk protein fractions except αs1-CN, with total CN, β-CN, and α-LA decreasing and κ-CN, β-LG, IgG, and LF increasing as parity increased (p < 0.05). (2) Mastitis significantly reduced the milk yield, fat percentage, protein percentage, and the contents of total CN, β-CN, κ-CN, and α-LA while increasing β-LG, IgG, and LF. (3) The SCS was negatively correlated with milk yield and α-LA but positively correlated with the fat percentage, protein percentage, κ-CN, β-LG, IgG, and LF. (4) When the DSCC increased to 50%, the milk yield decreased, while the milk protein percentage and κ-CN content significantly increased (p < 0.05). When the DSCC exceeded 50%, the fat percentage, protein percentage, total casein, αs1-CN, β-CN, κ-CN, β-LG, IgG, and LF decreased, while the α-LA content increased (p < 0.05). (5) When the PMN + LYMS increased, the milk yield and α-LA content rose, while the milk fat percentage, the milk protein percentage, and the contents of αs1-CN, β-CN, κ-CN, total CN, β-LG, IgG, and LF decreased (p < 0.05). Conversely, when the MACS increased, the milk yield and α-LA content declined, whereas the milk fat percentage, the milk protein percentage, and the contents of αs1-CN, β-CN, κ-CN, total CN, β-LG, IgG, and LF increased (p < 0.05). This study offers valuable insights into enhancing milk product quality, advancing the early diagnosis and mechanistic research of bovine mastitis, and the sustainable development of the dairy farming industry. Full article
(This article belongs to the Special Issue Sustainable Animal Production and Product Quality)
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16 pages, 3536 KiB  
Article
ATR Deficiency Impairs DNA Damage Repair and Accelerates Cellular Senescence in Bovine Mammary Epithelial Cells, Leading to Lactation Dysfunction
by Qijun Zhou, Zijian Geng, Shuai Lian, Jianfa Wang and Rui Wu
Animals 2025, 15(10), 1419; https://doi.org/10.3390/ani15101419 - 14 May 2025
Viewed by 441
Abstract
Mammary glands in cows are highly dynamic, making genomic stability particularly crucial. Continuous lactation and self-renewal of these glands are primary contributors to genomic instability. Results: We employed transcriptomic and proteomic methods to analyze the expressional characteristics in the mammary glands of cows [...] Read more.
Mammary glands in cows are highly dynamic, making genomic stability particularly crucial. Continuous lactation and self-renewal of these glands are primary contributors to genomic instability. Results: We employed transcriptomic and proteomic methods to analyze the expressional characteristics in the mammary glands of cows with varying levels of milk production. Our findings indicated differences in relevant pathways, including DNA damage repair and apoptosis, which are influenced by increasing parity. Notably, ATR protein levels in the mammary glands of low-yield dairy cows were reduced. Following in vitro silencing of ATR, β-galactosidase content increased in aging mammary epithelial cells, with cell cycle arrest in the G2 and S phases. Secretory phenotypes associated with aging, including IL-6, IL-10, IL-1β, INF-γ, and IL-2, were elevated, along with increased TNF-α content. The expressions of DNA repair-related proteins, including PIG3, PARP1, and Cleaved caspase3, were upregulated, and SP1 expression was decreased. Furthermore, the expressions of cytochrome C and BAK increased, and ATR silencing inhibited mTOR and STAT5 lactation signaling pathways, resulting in elevated STAT3 protein levels associated with mammary gland degeneration. Conclusions: This study emphasizes the significance of the ATR protein in the mammary glands of dairy cows, contributing valuable insights into maintaining their health and presenting a novel perspective on strategies to enhance their lifespan. Full article
(This article belongs to the Section Cattle)
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14 pages, 5014 KiB  
Article
UFBP1 Ameliorates Heat Stress-Induced Apoptosis via Mitochondria-Mediated Pathway in Bovine Mammary Epithelial Cells
by Yuan Li, Ran Yu, Shujing Tan, Yunlong Jiang, Longwei Sun, Manman Shen, Chuanjian Zhang, Kunlin Chen and Chengmin Li
Animals 2025, 15(9), 1233; https://doi.org/10.3390/ani15091233 - 27 Apr 2025
Viewed by 571
Abstract
Heat stress in dairy cows is aggravated by Global warming, which negatively affects their performance and health, especially high yielding cows are more susceptible to high temperature and humidity in summer. Besides increasing body temperature and reducing feed intake, heat stress also compromises [...] Read more.
Heat stress in dairy cows is aggravated by Global warming, which negatively affects their performance and health, especially high yielding cows are more susceptible to high temperature and humidity in summer. Besides increasing body temperature and reducing feed intake, heat stress also compromises mammary gland function by inducing apoptosis in bovine mammary epithelial cells (BMECs). UFBP1 (Ufm1-binding protein 1) serves as an essential component of ufmylation, is crucial for the preservation of cellular homeostasis. However, little is known about its contribution to heat stress-induced apoptosis in BMECs. Therefore, the present study aimed to elucidate the effect of UFBP1 on heat stress-induced apoptosis through knockdown and overexpression of UFBP1 in BMECs. The results showed that heat stress triggered cell apoptosis (increased apoptosis rate and Bax/Bcl-2 protein expression) and decreased the expression of genes associated with the production of milk fat and protein both in vivo and in vitro studies. Furthermore, UFBP1 silencing aggravated the high-temperature-induced cell damage, and overexpression of UFBP1 attenuated heat stress-induced mitochondrial dysfunction, as evidenced by increased mitochondrial membrane potential (MMP), ATP synthesis and NAD+/NADH ratio, as well as the reduced reactive oxygen species (ROS) generation. Importantly, the mitochondrial apoptosis pathway triggered by heat stress was blocked by UFBP1, as indicated by the reduced apoptosis rate and Bax/Bcl-2 protein expression. In addition, UFBP1 restored the expression of milk fat and protein-related genes in heat-stressed BMECs. In conclusion, these findings indicate that UFBP1 may serve as a promising therapeutic target for ameliorating heat stress in dairy cows, thereby providing novel theoretical insights into the mitigation of adverse thermal stress effects on livestock productivity. Full article
(This article belongs to the Special Issue Genetic Research for Improving Livestock Heat Stress Resistance)
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18 pages, 5034 KiB  
Article
Variations in Human Milk Metabolites After Gestational Diabetes: Associations with Infant Growth
by Alice Fradet, Line Berthiaume, Laurie-Anne Laroche, Camille Dugas, Julie Perron, Alain Doyen, Étienne Audet-Walsh and Julie Robitaille
Nutrients 2025, 17(9), 1466; https://doi.org/10.3390/nu17091466 - 26 Apr 2025
Viewed by 594
Abstract
Background/Objectives: Gestational diabetes mellitus (GDM) is a condition characterized by hyperglycemia and is associated with increased risk of obesity and diabetes in exposed children. Differences in human milk composition between women with (GDM+) and without GDM (GDM-) suggest that GDM could impact milk [...] Read more.
Background/Objectives: Gestational diabetes mellitus (GDM) is a condition characterized by hyperglycemia and is associated with increased risk of obesity and diabetes in exposed children. Differences in human milk composition between women with (GDM+) and without GDM (GDM-) suggest that GDM could impact milk production and composition, potentially influencing infant growth. However, this association remains poorly understood. The objective was to study the association between GDM and human milk composition and its influence on infant growth, focusing on metabolites and bioactive molecules involved in energy metabolism. Methods: Using a cross-sectional design, 24 metabolites were measured by GC-MS in human milk obtained at 2 months postpartum from 20 GDM+ women and 29 GDM- women. Anthropometric measures, as well as lipid and glycemic profiles, were collected. Infant weight and length data were obtained from health records. Results: Human milk metabolites significantly differ between GDM+ and GDM- mothers, with higher levels of myristic acid, glycerol, uracil, arachidonic acid, and cholesterol in GDM+ milk (p < 0.05). Specific human milk metabolites showed distinct correlations with maternal glycemic as well as infant growth, depending on GDM status. While maternal glycemia was associated with succinate and malate in all groups, maternal glycemia was specifically correlated with valine and glutamate in GDM+ mothers. Additionally, in GDM+ women, α-ketoglutarate and glycine were negatively correlated with infant growth. Conclusions: The results of this study suggest that GDM can influence the mother’s health beyond delivery, impacting the mammary gland biology with effects on the human milk composition. Further, correlations with infant growth suggest that GDM-dependent variations in milk composition potentially influence infant growth and metabolism. Full article
(This article belongs to the Special Issue Nutrition in Children's Growth and Development)
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16 pages, 3661 KiB  
Article
Mechanism of Astragalus Polysaccharide in Alleviating Bovine Mammary Fibrosis Through ROS/NLRP3 Inhibition and EMT Regulation
by Jiang Zhang, Kejiang Liu, Tingji Yang, Hongwei Duan, Longfei Xiao, Quanwei Zhang, Yong Zhang, Weitao Dong and Xingxu Zhao
Antioxidants 2025, 14(5), 503; https://doi.org/10.3390/antiox14050503 - 23 Apr 2025
Viewed by 761
Abstract
Mastitis in dairy cows, typically caused by bacterial infection, is a common inflammatory condition of the mammary tissue that leads to fibrosis, adversely affecting cow health, milk production, and dairy product quality. Astragalus polysaccharide (APS) has shown effectiveness in alleviating inflammation and fibrosis [...] Read more.
Mastitis in dairy cows, typically caused by bacterial infection, is a common inflammatory condition of the mammary tissue that leads to fibrosis, adversely affecting cow health, milk production, and dairy product quality. Astragalus polysaccharide (APS) has shown effectiveness in alleviating inflammation and fibrosis in various organs. The study employed lipopolysaccharide (LPS) to induce fibrotic conditions in two experimental systems: MAC-T bovine mammary epithelial cells and Kunming mouse models. Key parameters, including relative gene mRNA expression, protein levels, and reactive oxygen species (ROS) levels, were assessed using RT-qPCR, Western blotting (WB), and 2’,7’-Dichlorofluorescin diacetate (DCFH-DA) techniques, while histological analysis of mammary tissue was performed using H&E and Masson trichrome staining. Measuring malondialdehyde (MDA) levels, assessing the enzyme activities of catalase (CAT), and superoxide dismutase (SOD) were two methods of assessing oxidative stress. These methods were also tested in mouse mammary glands. APS significantly decreased ROS concentrations (p < 0.01), restored oxidative stress balance in mice (p < 0.05), and reduced fibrosis and inflammation, as demonstrated by histological observations and analysis. It also exerted regulatory effects on fibrosis markers (E-cadherin, N-cadherin, α-SMA) and inflammation markers (NLRP3, ASC, Caspase-1, IL-1β), as demonstrated by changes in their mRNA and protein expression. These findings endorse APS’s viability as an alternative therapeutic agent for mammary fibrosis therapy by demonstrating its ability to inhibit epithelial-mesenchymal transition (EMT) in vitro and mammary fibrosis in vivo, while also mitigating ROS production and reducing inflammation. Full article
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22 pages, 4556 KiB  
Article
Phloretin–Chitosan Nanoparticles and Tamoxifen: Synergistic Modulation of BRCA Genes and Enhanced Sensitization in Breast Cancer
by Abeer A. Ageeli and Sahera F. Mohamed
Chemistry 2025, 7(3), 68; https://doi.org/10.3390/chemistry7030068 - 22 Apr 2025
Viewed by 645
Abstract
This study aims to evaluate the therapeutic potential of phloretin–chitosan nanoparticles (Ph-ChNPs), alone and in combination with the anticancer drug tamoxifen, in modulating breast cancer markers and improving in vivo treatment outcomes. Ph-ChNPs were prepared by ionic gelation in the presence of Tripolyphosphate [...] Read more.
This study aims to evaluate the therapeutic potential of phloretin–chitosan nanoparticles (Ph-ChNPs), alone and in combination with the anticancer drug tamoxifen, in modulating breast cancer markers and improving in vivo treatment outcomes. Ph-ChNPs were prepared by ionic gelation in the presence of Tripolyphosphate (TPP) solution as a crosslinker agent. The nanoparticles were characterized using DLS, TEM, UV-VIS and FT-IR spectroscopy. In vitro cytotoxic assay of Ph-ChNPs on MCF-7 breast cancer cell lines revealed anticancer activity with an IC50 value of 32.12 ± 1.63 µg/mL. In vivo studies were carried out on mice, treated with DMBA to induce breast cancer and followed the effect of the prepared nanoparticle, either alone or with combination with tamoxifen, on mice health. The biochemical parameters measured after treatment with Ph-ChNPs alone showed an improvement in lipid profile with decreased total cholesterol (TC) and Triglyceride (TG) levels and increased HDL-c levels. Ph-ChNPs significantly reduced IL-6 and cyclin D1 levels, with a slight increase in cyclin E2 levels. Antioxidant enzyme levels were improved, and oxidative stress markers were reduced. The combination treatment showed a synergistic effect in reducing inflammation and cell proliferation. DMBA-injected mice had substantially increased BRCA1 and BRCA2 gene expression. Ph-ChNP-treated mice showed well-organized mammary gland structures, while DMBA-injected mice displayed dense tumor cell aggregations. Ph-ChNPs and tamoxifen treatments improved histopathological variations, with the combination treatment showing significant apoptosis of tumor cells. This study demonstrates the significant potential of Ph-ChNPs combined with tamoxifen in breast cancer treatment. The combination therapy effectively reduces tumor growth, induces apoptosis and modulates critical breast cancer markers, offering a promising therapeutic strategy. Full article
(This article belongs to the Section Medicinal Chemistry)
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28 pages, 4968 KiB  
Article
Peptide Profiling in Dairy Cow Dry Secretions: Temporal Changes and Comparative Analysis Between Healthy and Subclinical Mastitis Cows
by Barjam Hasanllari, Benjamin P. Willing, Liang Li, Xian Luo and Burim N. Ametaj
Dairy 2025, 6(2), 19; https://doi.org/10.3390/dairy6020019 - 15 Apr 2025
Viewed by 1165
Abstract
The dairy industry relies on the health and well-being of dairy cows for the optimal production of milk and dairy products. Mastitis, a prevalent and economically burdensome disease characterized by udder inflammation, poses significant challenges to dairy farmers worldwide. In this study, we [...] Read more.
The dairy industry relies on the health and well-being of dairy cows for the optimal production of milk and dairy products. Mastitis, a prevalent and economically burdensome disease characterized by udder inflammation, poses significant challenges to dairy farmers worldwide. In this study, we employed peptidomics to explore the peptide profiles of dry secretions collected from dairy cows at specific intervals during the dry-off period. We hypothesized that alterations in peptide composition during the dry period may influence pathogen proliferation and immune cell functioning, thereby impacting mastitis susceptibility. Our objectives were to investigate the following: (i) differences in peptide composition and alterations between healthy cows and those with subclinical mastitis, potentially serving as biomarkers for early mastitis detection and offering insights into udder bioprocesses; (ii) variations in peptide profiles between the early (day 2) and later (day 21) stages of the dry-off period during both health conditions. Dry secretions were collected from 16 udder quarters of 8 cows at two defined time points—Day 2 (D2) and Day 21 (D21)—during the dry period. Our results revealed distinct peptide patterns between healthy and subclinical mastitis cows, as well as temporal variations in peptide profiles throughout the dry-off period. A total of 1235 peptides, originating from 59 distinct proteins (primarily β-casein), were detected across the four groups: subclinical mastitis day 21 (SCM-D21), subclinical mastitis day 2 (SCM-D2), healthy day 21 (H-D21), and healthy day 2 (H-D2). Furthermore, 56 out of the 1235 peptides identified in total matched known functional peptides, with a total of 17 different functions including antimicrobial, antioxidant, and immunomodulatory, suggesting their potential roles in mastitis pathogenesis and mammary gland physiology. Comparative analyses revealed changes in the levels of these functional peptides across the four different groups, suggesting their potential roles in regulating immune responses, oxidative stress, inflammatory processes, and other biological activities during subclinical mastitis and the dry-off period. These findings provide valuable insights into mastitis detection, management strategies, and dairy cow health monitoring, offering promising avenues for enhancing milk quality and dairy industry sustainability. Full article
(This article belongs to the Section Dairy Systems Biology)
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25 pages, 1707 KiB  
Article
The Genistein Supply and Elemental Composition of Rat Kidneys in an Induced Breast Cancer Model
by Dorota Skrajnowska, Arkadiusz Szterk, Karol Ofiara, Paweł Kowalczyk and Barbara Bobrowska-Korczak
Nutrients 2025, 17(7), 1184; https://doi.org/10.3390/nu17071184 - 28 Mar 2025
Viewed by 735
Abstract
Background: Many natural phytochemicals support the work of the kidneys. The health effects of genistein have been confirmed in many kidney diseases (inflammation and acute kidney injury, cancer or menopausal or senile changes). Genistein through various mechanisms can affect kidney conditions. Objectives: The [...] Read more.
Background: Many natural phytochemicals support the work of the kidneys. The health effects of genistein have been confirmed in many kidney diseases (inflammation and acute kidney injury, cancer or menopausal or senile changes). Genistein through various mechanisms can affect kidney conditions. Objectives: The purpose of this work was to analyze the supply of various forms of genistein at a low dose (0.2 mg/kg b.w.) on the renal mineral composition of rats under conditions of mammary gland tumorigenesis (induced with DMBA). Methods: Sprague rats at the age of 40 days were divided into four research groups, i.e., a control group receiving only standard feed and four groups receiving feed supplemented with genistein in the form of nanoparticles (0.1 mg/mL, i.e., 0.2 mg/kg.i.d.) (size: 92 ± 41 nm), genistein in microparticle form (0.1 mg/mL, i.e., 0.2 mg/kg.i.d.) (size: 587 ± 83 nm) and genistein in macroparticle form (normal, classical) (0.1 mg/mL, i.e., 0.2 mg/kg.i.d.). Mammary gland cancer was induced using DMBA (7,12-dimethyl-1,2-benz(a)anthracene). The experiment lasted 100 days. The concentrations of Ca, Zn, Fe, Cu, As, Se, Rb, Sr, Mo, B, and Mn were measured using the ICP-MS method, while the levels of K, Mg, and Na were measured using the FAAS method. Results: It was shown that, depending on the degree of miniaturization of genistein, its administration affected changes in kidney mineral composition, primarily resulting in a strongly reduced calcium content in the group of rats receiving nanogenistein. We found a negative impact of nanogenistein administration on the amount of calcium and iron, indicating an increased distribution or excretion of these elements from the body, as well as an increase in the number of elements, especially magnesium, sodium, zinc, boron, and copper concentrations, compared to the non-supplemented group. Conclusions: This study confirms the need for thorough clinical analyses in the future, with regard to the effects of genistein, especially its nanoforms on the body. Full article
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