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18 pages, 2564 KB  
Article
The Effect of β-Hydroxy-β-Methyl Butyrate (HMB) upon Acute Fed-State Muscle Protein Turnover in Older Men and Women: A Randomized Double-Blind Controlled Crossover Clinical Trial
by Kenneth Smith, Haitham Abdullah, Supreeth Rudrappa, Amanda Gates, Jonathan Lewis, Iskandar Idris, Joseph J. Bass, Hannah Crossland, Daniel J. Wilkinson, Min Tian, Deborah S. Hustead, Geraldine E. Baggs, Suzette L. Pereira, Bethan E. Phillips and Philip J. Atherton
Nutrients 2026, 18(9), 1449; https://doi.org/10.3390/nu18091449 - 30 Apr 2026
Abstract
Background/Objectives: Anabolic resistance is thought to underlie muscle loss in sarcopenia. Here, we investigated the adjuvant role of beta-hydroxy-beta-methylbutyrate (HMB), a leucine metabolite, on the acute muscle anabolic response to oral protein supplementation in older adults. Methods: A total of 24 [...] Read more.
Background/Objectives: Anabolic resistance is thought to underlie muscle loss in sarcopenia. Here, we investigated the adjuvant role of beta-hydroxy-beta-methylbutyrate (HMB), a leucine metabolite, on the acute muscle anabolic response to oral protein supplementation in older adults. Methods: A total of 24 community-dwelling older adults (68.5 ± 0.6 years; 13 men, 11 women) were randomized in a cross-over double-blind design to 40 g whey protein (Control) or 40 g whey protein with 3 g calcium–HMB (HMB). Subjects received a primed constant infusion of 13C6 phenylalanine to assess muscle protein synthesis (MPS, by tracer incorporation in myofibrils) and muscle protein breakdown (MPB, via arterio-venous dilution) at baseline and post supplementation. Fasted and 3 h fed-state plasma HMB, aminoacidemia, rates of MPS, MPB, limb and muscle blood flow were measured. Results: In all subjects, both interventions displayed significant increases in MPS in response to feeding [fasted to 3 h-fed change (mean ± SEM, standard error of the mean). Males: control, +0.032 ± 0.006%.h−1; HMB, +0.023 ± 0.004%.h−1; females: control, +0.023 ± 0.006%.h−1; HMB, +0.038 ± 0.006%.h−1, p < 0.05]. In older women, the addition of HMB further enhanced the MPS response (fasted to 3 h-fed change, p = 0.0495) and area under the curve (p = 0.0364) versus protein alone. During the late-fed period, MPB significantly decreased in HMB versus control (p = 0.0298), and this was also observed when subjects were separated by sex (p = 0.0012). Conclusions: High-dose protein bolus feeding increased MPS in older adults. Surprisingly, 40 g whey did not maximize the anabolic response in older women, and HMB further increased the MPS feeding response to protein. HMB further suppressed the MPB feeding response over a longer period of time. Further work is needed to understand the apparent sexual dimorphic MPS response to high protein. Full article
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21 pages, 8696 KB  
Article
Homocysteine Drives Hippocampal Blood–Brain Barrier Disruption and Cognitive Decline Under Chronic Stress via DNA Hypomethylation of Cav1.2
by Mao-Yang Zhou, Jin-Shan Li, Zhao-Xin Sun, Jie Yin, Yun Zhao, Fang Xie, Xue Wang, Sheng-Hui Zhang, Zhao-Wei Sun and Ling-Jia Qian
Brain Sci. 2026, 16(5), 491; https://doi.org/10.3390/brainsci16050491 - 30 Apr 2026
Abstract
Background: Chronic stress is a major risk factor for cognitive decline and blood–brain barrier (BBB) disruption, yet the underlying molecular mechanisms remain elusive. This study aimed to investigate the specific role of the metabolic intermediate homocysteine (Hcy) in chronic stress-induced BBB dysfunction and [...] Read more.
Background: Chronic stress is a major risk factor for cognitive decline and blood–brain barrier (BBB) disruption, yet the underlying molecular mechanisms remain elusive. This study aimed to investigate the specific role of the metabolic intermediate homocysteine (Hcy) in chronic stress-induced BBB dysfunction and cognitive impairment. Methods: We utilized a male Sprague-Dawley rat model of chronic unpredictable mild stress (CUMS) and administered vitamin B complex to lower Hcy levels in vivo. Regional Hcy accumulation, BBB permeability, and cognitive behaviors were assessed. In vitro, primary rat brain microvascular endothelial cells (BMECs) were exposed to Hcy to evaluate barrier-forming function, transcriptomic alterations, DNA methylation patterns, Cav1.2 expression, and reactive oxygen species (ROS) production. Results: CUMS selectively induced BBB hyperpermeability and significant Hcy accumulation predominantly within the rat hippocampus, which correlated intimately with cognitive deficits. Lowering Hcy levels via vitamin B supplementation successfully restored hippocampal BBB integrity and alleviated cognitive impairment. In addition, elevated Hcy severely impaired the barrier function of BMECs. Mechanistically, Hcy reduced global DNA methylation in BMECs and specifically induced targeted DNA hypomethylation at the intro region of Cacna1c. This epigenetic shift caused the transcriptional derepression and overexpression of the Cav1.2 calcium channel. Upregulated Cav1.2 subsequently triggered a robust ROS burst, leading to tight junction degradation. Conclusions: Our findings unveil a novel metabolic–epigenetic axis where Hcy-driven Cacna1c hypomethylation directly disrupts BMECs function to dismantle the hippocampal BBB. Lowering Hcy or targeting this Hcy-Cav1.2 pathway establishes a promising therapeutic strategy for mitigating stress-related neurovascular damage and cognitive disorders. Full article
(This article belongs to the Section Neuropharmacology and Neuropathology)
20 pages, 3091 KB  
Article
Effects of 25-Hydroxyvitamin D3 Combined with Phytase and Probiotic on Calcium–Phosphorus Metabolism, Bone Development, and Growth Performance in Weaned Piglets
by Baoshi Shi, Saiming Gong, Jingjing Wang, Yuyue Xi, Zhiru Tang, Jingchun Gao, Yetong Xu and Zhihong Sun
Nutrients 2026, 18(9), 1428; https://doi.org/10.3390/nu18091428 - 30 Apr 2026
Abstract
Background/Objectives: Calcium–phosphorus metabolism is critical for skeletal development in weaned piglets. This study evaluated the effects of dietary 25-hydroxyvitamin D3 (25-OH-VD3) in combination with phytase and probiotics on mineral metabolism, bone development, and related molecular mechanisms in weaned piglets. Methods: [...] Read more.
Background/Objectives: Calcium–phosphorus metabolism is critical for skeletal development in weaned piglets. This study evaluated the effects of dietary 25-hydroxyvitamin D3 (25-OH-VD3) in combination with phytase and probiotics on mineral metabolism, bone development, and related molecular mechanisms in weaned piglets. Methods: Sixty 28-day-old weaned piglets (7.1 ± 1.30 kg) were randomly assigned to four dietary treatments for 31 days (including 3 days of acclimation): CON (basal diet + 50 µg/kg 25-OH-VD3), HI (CON + 50 mg/kg phytase), CY (CON +10 mg/kg probiotics), HICY (CON + 50 mg/kg phytase + 10 mg/kg probiotics). Apparent calcium digestibility, serum biochemical indices, bone mineral density (BMD), and mRNA and protein expression of calcium–phosphorus transport- and metabolism-related genes in jejunal mucosa and kidney were assessed. Results: Compared with CON, piglets in the HI, CY, and HICY groups showed higher apparent calcium digestibility (p < 0.05). Serum transforming growth factor-β was elevated in CY and HICY (p < 0.05). HI enhanced metatarsal and toe BMD (p < 0.05) and upregulated jejunal solute carrier family 34, member 2 (SLC34A2) and SLC34A3 mRNA expression (p < 0.05). In contrast, HICY reduced mRNA expression of transient receptor potential cation channel subfamily V member 6 and calcium-binding protein D28k, as well as of calcium-binding protein D9k and cytochrome P450 27B1 in the kidney (p < 0.05). Renal calcium-sensing receptor protein abundance increased in CY (p < 0.05). Conclusions: Supplementation of 25-OH-VD3 with phytase and/or probiotics improved calcium utilization and modulated key transport pathways, contributing to enhanced bone development in weaned piglets. These findings highlight coordinated nutritional regulation of mineral metabolism during early post-weaning growth. Full article
(This article belongs to the Special Issue Bone-Health-Promoting Bioactive Nutrition)
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12 pages, 759 KB  
Article
Evaluation of an In-House Developed Foot-and-Mouth Disease Virus SAT 3 Vaccine Strain Based on Antigen Productivity and Inactivation Kinetics for Commercial Feasibility
by Jae Young Kim, Sun Young Park, Gyeongmin Lee, Giyoun Cho, Seung-A Hwangbo, Jong-Hyeon Park and Young-Joon Ko
Vaccines 2026, 14(5), 381; https://doi.org/10.3390/vaccines14050381 - 24 Apr 2026
Viewed by 210
Abstract
Background: In the Republic of Korea, a bivalent foot-and-mouth disease (FMD) vaccine covering serotypes O and A is administered to livestock, while antigens for the other serotypes are stockpiled in overseas antigen banks. To achieve self-reliance in FMD vaccine production, various vaccine strains [...] Read more.
Background: In the Republic of Korea, a bivalent foot-and-mouth disease (FMD) vaccine covering serotypes O and A is administered to livestock, while antigens for the other serotypes are stockpiled in overseas antigen banks. To achieve self-reliance in FMD vaccine production, various vaccine strains have been developed using in-house technology. Although SAT 3 has historically been confined largely to Africa, preparedness against this serotype remains necessary, as the possibility of its introduction into Korea cannot be completely excluded. Methods: In this regard, we evaluated the commercial potential of the SAT 3 ZIM-R vaccine strain by assessing antigen productivity, scalability, inactivation kinetics, and immunogenicity. Results: Supplementation with 3 mM Ca2+ markedly increased antigen yield compared with that obtained in the absence of calcium. Further optimization showed that antigen yield was highest at pH 8.0–8.5. During scale-up, antigen yield was maintained at 9.2–9.8 μg/mL in flask cultures and remained high at approximately 7.8 μg/mL in a bioreactor, demonstrating robust scalability. Treatment with 2 mM binary ethylenimine at 26 °C achieved complete inactivation within 24 h. Vaccines formulated with the SAT 3 ZIM-R antigen produced either in flasks or in a bioreactor induced comparable neutralizing antibody responses in pigs following both the primary and booster immunizations. Conclusions: Collectively, these findings indicate that SAT 3 ZIM-R is a promising vaccine candidate for large-scale vaccine antigen production and the future establishment of a domestic FMD antigen bank in Korea. Full article
(This article belongs to the Section Veterinary Vaccines)
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0 pages, 344 KB  
Article
Nutrient Intake and Physical Activity of School-Aged Children with Trisomy 21 Living in Manitoba, Canada
by Maria S. Baranowski, Carla G. Taylor, Nancy Hansen and Shahin Shooshtari
Nutrients 2026, 18(9), 1330; https://doi.org/10.3390/nu18091330 - 23 Apr 2026
Viewed by 252
Abstract
Background: Children and adults with Trisomy 21 are more likely to develop nutrition-related conditions and diseases. The nutrition-related health of Canadians with Trisomy 21 is unknown. We aimed to determine the nutrient intake and physical activity of school-aged children with Trisomy 21 in [...] Read more.
Background: Children and adults with Trisomy 21 are more likely to develop nutrition-related conditions and diseases. The nutrition-related health of Canadians with Trisomy 21 is unknown. We aimed to determine the nutrient intake and physical activity of school-aged children with Trisomy 21 in Manitoba, Canada. Methods: Mothers of 14 school-aged children (n = 7 female, average age 9 years old) with Trisomy 21 completed a 24 h dietary recall and a survey that included questions about their children’s nutrition and physical activity. Nutrient intake analysis was conducted to compare food and beverage consumption with dietary guidelines and nutrient recommendations. Data were analyzed descriptively. Results: Most children with T21 included in this study consumed an adequate average intake of daily protein, carbohydrate, and iron; an inadequate average intake of daily dietary fibre and calcium; and an excessive average daily intake of added sugars and saturated fat. Notably, all children consumed inadequate vitamin D and excessive sodium. Most children consumed a dietary supplement (10/14), engaged in moderate-intensity physical activity (10/14), and were active for more than 60 min per day (12/14). Conclusions: Most children with Trisomy 21 included in this study met daily physical activity recommendations. However, despite a variety of foods reportedly consumed across all food groups, nutrient intake among school-aged children with Trisomy 21 included in this study was mixed, as both deficiencies and excessive amounts of some nutrients were observed. There is a need to improve the nutrient intake of children with Trisomy 21 to reduce their risk of developing nutrition-related conditions and diseases. Full article
(This article belongs to the Special Issue Nutrition in Children's Growth and Development: 2nd Edition)
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15 pages, 3040 KB  
Article
Impact of Clinoptilolite and Anionic Salts on Calcium Homeostasis, Parathyroid Hormone, and Related Metabolic Parameters in Periparturient Dairy Cows
by Pengyu Huang, Xiu Su, Yuanyin Guo, Chong Ma and Jie Cao
Vet. Sci. 2026, 13(5), 408; https://doi.org/10.3390/vetsci13050408 - 22 Apr 2026
Viewed by 234
Abstract
This study compared the regulatory effects of dietary supplementation with natural clinoptilolite (CLN) versus a dietary cation-anion difference (DCAD) regimen on calcium homeostasis in dairy cows during the last 21 days prepartum. Results showed that cows in the DCAD group exhibited significantly higher [...] Read more.
This study compared the regulatory effects of dietary supplementation with natural clinoptilolite (CLN) versus a dietary cation-anion difference (DCAD) regimen on calcium homeostasis in dairy cows during the last 21 days prepartum. Results showed that cows in the DCAD group exhibited significantly higher blood ionized calcium (iCa) and parathyroid hormone (PTH) concentrations than those in the CLN group (p < 0.05). Serum PTH concentrations displayed a declining trend in both groups prepartum, which deviates from the classical theory of compensatory PTH secretion, suggesting that alternative compensatory pathways may be involved in the regulation of calcium homeostasis during the periparturient period in dairy cows. Monitoring of calcium homeostasis and related metabolic parameters following postpartum oral calcium bolus administration revealed that the incidence of subclinical hypocalcemia in the DCAD group was 26%, lower than the 62% observed in the CLN group. However, blood iCa concentrations returned to normal levels more rapidly in the CLN group. Additionally, CLN supplementation was associated with more stable blood pH and lower prepartum serum potassium concentrations (p < 0.05) that remained within the physiological range, which may contribute to improved tissue sensitivity to PTH. Full article
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25 pages, 3720 KB  
Article
Cryogenic Damage and Trehalose Protection in Culter alburnus Sperm: An Integrated Assessment of Quality, Physiology, and Protein Expression
by Shun Cheng, Shi-Li Liu, Mei-Li Chi, Wen-Ping Jiang, Jian-Bo Zheng, Chao Zhu, Jun-Zhi Luo and Fei Li
Animals 2026, 16(8), 1245; https://doi.org/10.3390/ani16081245 - 18 Apr 2026
Viewed by 254
Abstract
To address cryodamage in Culter alburnus sperm, this study evaluated the effects of trehalose supplementation in a conventional cryomedium (D-15 + 10% ethylene glycol). Six experimental groups were established: fresh sperm (G1), a conventional cryomedium (G2), groups supplemented with 10, 100, or 200 [...] Read more.
To address cryodamage in Culter alburnus sperm, this study evaluated the effects of trehalose supplementation in a conventional cryomedium (D-15 + 10% ethylene glycol). Six experimental groups were established: fresh sperm (G1), a conventional cryomedium (G2), groups supplemented with 10, 100, or 200 mmol/L trehalose (G3–G5), and a control group with extender only (G6). The group with 100 mmol/L trehalose (G4) was associated with improved post-thaw motility parameters (activation rate, movement time, and lifespan) and higher antioxidant (superoxide dismutase and catalase) and energy metabolism (ATPase, succinate dehydrogenase, lactate dehydrogenase) enzyme activities. Ultrastructural damage in G4 included partial plasma membrane rupture and mitochondrial swelling, while G6 exhibited additional damage features including membrane disintegration, mitochondrial disruption, and flagellar fracture. Proteomic analysis revealed that, compared to G1, G4 exhibited higher abundance of proteins (e.g., Histone H2A, cytochrome c oxidase, profilin) involved in structural integrity and energy homeostasis, whereas G6 showed signatures of oxidative stress and metabolic dysfunction (lower abundance of NADH dehydrogenase and higher abundance of calcium-transporting ATPase and glutathione S-transferase). In conclusion, 100 mmol/L trehalose was associated with improved cryopreservation outcomes, and the proteins identified provide a basis for further investigation. This approach offers a framework for refining germplasm conservation strategies in aquaculture. Full article
(This article belongs to the Section Aquatic Animals)
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21 pages, 1965 KB  
Article
Impact of Monofloral Pollen Diets on the Development of Hypopharyngeal Glands and Modulation of Enzymatic, Non-Enzymatic, and Ionic Biomarker Activities in Selected Fat Body Segments and Hemolymph of Apis mellifera Workers
by Maciej Sylwester Bryś, Krzysztof Olszewski, Bernard Staniec, Patrycja Staniszewska and Aneta Strachecka
Molecules 2026, 31(8), 1315; https://doi.org/10.3390/molecules31081315 - 17 Apr 2026
Viewed by 533
Abstract
The homogenization of landscapes and reduction in floral diversity have increasingly led to low diversity in pollen diets for honey bees. In this study, we examined the effects of monofloral pollen diets based on wind-pollinated (Corylus sp., Pinus sp.) and insect-pollinated plants [...] Read more.
The homogenization of landscapes and reduction in floral diversity have increasingly led to low diversity in pollen diets for honey bees. In this study, we examined the effects of monofloral pollen diets based on wind-pollinated (Corylus sp., Pinus sp.) and insect-pollinated plants (Brassica napus L., Phacelia sp., Solidago sp., Fagopyrum sp.) on the development of hypopharyngeal glands (HPGs), activities of enzymatic (AST, ALT, ALP, GGTP) and non-enzymatic (urea, uric acid) biomarkers, as well as magnesium, calcium, and phosphorus concentrations in the hemolymph and fat bodies from different locations (tergite 3, 5 and sternite) in worker bees. Even a small (10%) addition of pollen to sugar candy stimulated the development of acini compared to the control group, with phacelia, buckwheat, and goldenrod pollen having the strongest effects. The largest acini developed in the 14-day-old bees fed with Phacelia pollen, whereas the collecting duct diameters were significantly reduced in all the pollen-supplemented groups. Enzymatic biomarker activities were the highest in the hemolymph of newly emerged bees and increased with age across all the tissues, particularly in tergite 5, with the highest activities recorded in the bees fed with insect-pollinated plant pollen. Non-enzymatic parameters and ion concentrations also varied with tissue type and segmental location, generally increasing with age and reaching the highest values in tergite 5. Regardless of the tissue (the fat body vs. hemolymph), the bees fed a diet containing pollen from Brassica napus, Phacelia, Solidago, and Fagopyrum had higher concentrations of calcium, magnesium, and phosphorus, while the effects of hazel and pine pollen were less pronounced. These findings suggest that even limited pollen supplementation can positively shape the morphological and biochemical physiology of worker bees. Understanding these relationships is crucial for developing strategies to support bee health under increasing environmental pressures and changing floral availability. Full article
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15 pages, 247 KB  
Article
Oyster Shell Powder as a Calcium-Based Buffer for Stabilizing Acidic Fruit and Vegetable Waste During Black Soldier Fly Larvae Bioconversion
by Nhien Thi Nguyen and Nam Hoang Tran
Sustainability 2026, 18(8), 3949; https://doi.org/10.3390/su18083949 - 16 Apr 2026
Viewed by 346
Abstract
The rapid accumulation of fruit and vegetable waste and oyster shell residues presents increasing environmental challenges, particularly in regions with intensive agricultural and aquaculture production. This study evaluated the use of oyster shell powder as a calcium-based buffering additive to stabilize acidic fruit [...] Read more.
The rapid accumulation of fruit and vegetable waste and oyster shell residues presents increasing environmental challenges, particularly in regions with intensive agricultural and aquaculture production. This study evaluated the use of oyster shell powder as a calcium-based buffering additive to stabilize acidic fruit and vegetable waste during rearing of black soldier fly larvae (BSFL, Hermetia illucens). Five substrates containing 0, 2, 4, 6, and 8% oyster shell powder (fresh weight basis) were prepared and used for larval rearing under controlled conditions for 12 days. Substrate pH dynamics, larval growth performance, substrate utilization efficiency, and nutritional composition of larval biomass were assessed. Oyster shell supplementation significantly increased substrate pH in a dose-dependent manner, shifting the substrate from strongly acidic conditions toward the range favorable for larval development. Moderate supplementation levels (2–4%) resulted in the highest larval biomass, survival rate, dry matter reduction, and bioconversion efficiency, whereas excessive supplementation reduced performance. Protein content increased at moderate supplementation levels, while lipid content decreased with higher oyster shell inclusion. Calcium concentration in larval biomass increased proportionally with supplementation, whereas essential amino acid composition remained stable. These results demonstrate that oyster shell powder can be used as an effective buffering material to improve the stability of acidic organic waste substrates and enhance BSFL-based bioconversion. The combined utilization of fruit and vegetable waste and oyster shell residues represents a practical approach for integrated waste valorization and supports the development of circular bioeconomy strategies for sustainable protein production. Full article
16 pages, 411 KB  
Article
Dietary Tartary Buckwheat Flavonoids Enhance Antioxidant Capacity and Regulate Lipid Metabolism via the AMPK Pathway in Liangshan Yanying Chickens
by Dongdong Li, Yi Zhang, Anqiang Lai, Binlong Chen, Silu Wang, Caiyun Sun, Zhiqiu Huang and Zengwen Huang
Vet. Sci. 2026, 13(4), 375; https://doi.org/10.3390/vetsci13040375 - 13 Apr 2026
Viewed by 314
Abstract
The present work was designed to investigate the impacts of tartary buckwheat flavonoids (TBF) on the growth performance and physiological metabolism of Liangshan Yanying chickens. A total of 144 healthy 4-week-old Liangshan Yanying chickens of uniform body weight were randomly divided into four [...] Read more.
The present work was designed to investigate the impacts of tartary buckwheat flavonoids (TBF) on the growth performance and physiological metabolism of Liangshan Yanying chickens. A total of 144 healthy 4-week-old Liangshan Yanying chickens of uniform body weight were randomly divided into four groups. Each group consisted of six replicates with six chickens per replicate. The control group was fed a basal diet, whereas the treatment groups received the same basal formulation supplemented with TBF at concentrations of 20, 40, and 60 mg/kg, respectively. The entire feeding trial lasted for 10 consecutive weeks. Growth performance, serum parameters, bone quality, slaughter traits, and hepatic lipid metabolism were determined and statistically analyzed. Results showed that dietary TBF supplementation had no significant impact on the overall growth performance (p > 0.05); however, final body weight and average daily weight gain displayed a positive linear trend in response to increasing TBF levels (0.05 < p < 0.1). For serum parameters, TBF supplementation significantly increased total antioxidant capacity (p < 0.05) and decreased malondialdehyde content (p < 0.05) in a linear manner. Specifically, compared with the control group, the 60 mg/kg TBF group increased T-AOC by approximately 64.6% and reduced MDA by approximately 67.9%, demonstrating a robust antioxidant effect. A linear increase in high-density lipoprotein cholesterol levels approaching significance (p = 0.055) was also observed. A significant quadratic regulatory effect of TBF was observed on serum glucose concentrations (p < 0.05), whereby the 60 mg/kg TBF dose reduced serum glucose by 15.6% relative to the control (p < 0.05), reflecting a robust hypoglycemic effect. Regarding bone quality, supplementation with 20 and 60 mg/kg TBF significantly elevated tibial phosphorus content relative to the 0 mg/kg TBF group (p < 0.05). Regression analysis showed that increasing dietary TBF levels linearly elevated tibial calcium content (p < 0.05) and resulted in linear increasing trends in tibial ash content, tibial phosphorus content, femur ash content, and femur calcium content (0.05 < p < 0.1). Concerning slaughter performance, dietary TBF inclusion resulted in a significant linear rise in breast muscle percentage (p < 0.05). At the molecular level, TBF upregulated the mRNA expression of hepatic AMPKα1 and CPT1, while concurrently downregulating the expression of FAS and ACC (p < 0.05). Collectively, dietary TBF supplementation in Liangshan Yanying chickens effectively improved antioxidant capacity, promoted tibial calcium and phosphorus deposition, regulated hepatic fatty acid oxidation and synthesis via AMPK-related genes, and enhanced lean meat deposition, with no adverse effects on growth performance under the experimental conditions. Full article
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18 pages, 2929 KB  
Article
In Vitro Analysis of Gene and Protein Expression in Primary Limbal Epithelial Cells Exposed to Differentiation-Inducing Medium
by Shweta Suiwal, Virendra Kumar, Tanja Stachon, Priya Katiyar, Fabian N. Fries, Berthold Seitz, Shuailin Li, Shao-Lun Hsu, Shanhe Liu, Swarnali Kundu, Maryam Amini, Sabrina Häcker and Nóra Szentmáry
Biology 2026, 15(8), 610; https://doi.org/10.3390/biology15080610 - 12 Apr 2026
Viewed by 496
Abstract
Purpose: To study the time course of the differentiation process and its regulatory networks in primary limbal epithelial cells (pLECs) using serum-free, low calcium Keratocyte growth medium 3 (KGM3) and CnT-2D differentiation medium. Methods: pLECs were isolated from corneoscleral rims from healthy [...] Read more.
Purpose: To study the time course of the differentiation process and its regulatory networks in primary limbal epithelial cells (pLECs) using serum-free, low calcium Keratocyte growth medium 3 (KGM3) and CnT-2D differentiation medium. Methods: pLECs were isolated from corneoscleral rims from healthy donors and cultured in serum-free low calcium (0.06 mM Ca2+) KGM3. Differentiation was induced by supplementation with CnT-2D differentiation medium, while control cells were maintained in low-calcium KGM3 medium. Gene and protein expression analyses were performed using qPCR and Western blotting, respectively, at 72 h and at 5, 7, 10, and 14 days post-supplementation to determine the optimal time course of differentiation induction. Results: CnT-2D differentiation medium supplementation resulted in a significant upregulation of differentiation-associated markers, including desmoglein 1 (DSG1), paired box domain 6 (PAX6), keratin 3 (KRT3), fatty acid binding protein 5 (FABP5), cellular retinoic acid binding protein 2 (CRABP2), alcohol dehydrogenase 7 (ADH7), aldehyde dehydrogenase 1A1 (ALDH1A1), with the most pronounced changes observed at day 10 post-supplementation (p ≤ 0.05). Conclusions: CnT-2D differentiation medium effectively initiates differentiation of limbal epithelial cells in vitro. The gradual increase in the expression of key differentiation markers, including DSG1, KRT3, and PAX6, indicates that CnT-2D medium successfully induces differentiation in 2D cultured primary limbal epithelial cells. However, subcellular localization of these markers, epithelial barrier function, and differentiation in 3D models were not assessed and remain to be investigated. Full article
(This article belongs to the Section Cell Biology)
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17 pages, 1039 KB  
Article
Effects of Astaxanthin as a Feed Additive on Growth Performance, Intestinal Microbiota and Clinical Parameters in Preweaning Female Holstein Calves: A Preliminary Study
by Elena Scaglia, Valeria Sergi, Laura Giagnoni, Livio Galosi, Anna Simonetto, Giulia Ferronato, Gianni Gilioli and Valentina Caprarulo
Animals 2026, 16(8), 1173; https://doi.org/10.3390/ani16081173 - 11 Apr 2026
Viewed by 433
Abstract
The preweaning period is a critical phase for dairy calves, during which gastrointestinal disorders, particularly diarrhea, remain a major cause of morbidity and antimicrobial use. Astaxanthin, a xanthophyll carotenoid with antioxidant and anti-inflammatory properties, has shown potential to support intestinal health in several [...] Read more.
The preweaning period is a critical phase for dairy calves, during which gastrointestinal disorders, particularly diarrhea, remain a major cause of morbidity and antimicrobial use. Astaxanthin, a xanthophyll carotenoid with antioxidant and anti-inflammatory properties, has shown potential to support intestinal health in several animal species, but information in preweaning dairy calves is limited. The objective of this study was to evaluate the effects of astaxanthin supplementation administered through milk replacer on growth performance, clinical health, metabolic profile, and fecal microbiota in preweaning Holstein calves. Twenty-four female Holstein calves (body weight, mean ± SD: 49.51 ± 12.14 kg) were randomly assigned to a control group (CTR; n = 12) or an astaxanthin-supplemented group (TRT; n = 12). Treated calves received 40 mg/d of astaxanthin from week 0 to 4 and 80 mg/d from week 4 to 8. Body weight, feed intake, rectal temperature, and fecal consistency score, fecal color score and clinical scores were recorded throughout the trial. Blood samples were collected in weeks 0 and 8 for metabolic profiling, and fecal samples were collected in weeks 0, 4, and 8 to assess fecal consistency and bacterial populations. Astaxanthin supplementation did not affect body weight, average daily gain, or overall feed intake. However, treated calves exhibited a lower frequency of abnormal fecal consistency scores, indicating reduced diarrhea incidence compared with control calves. Fecal microbiological analysis revealed lower coliform counts in astaxanthin-supplemented calves in weeks 4 and 8, whereas total bacterial counts were greater in week 8. Most blood metabolites were primarily influenced by age-related physiological changes; however, circulating calcium concentrations were greater in treated calves. These results suggest that astaxanthin may represent a promising nutritional strategy to support gastrointestinal health during early life, although larger studies are needed to confirm these findings. Full article
(This article belongs to the Special Issue Diarrhea in Neonatal Ruminant Calves: Diagnosis and Treatment)
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19 pages, 846 KB  
Article
The Effects of a Phytochemical Supplement Blend on Markers of Exercise-Induced Muscle Damage: A Randomised Controlled Trial
by Josh Thorley, Kirsty M. Reynolds, Matt Nickels, Stephen J. Bailey, Ronald Kingma and Tom Clifford
Nutrients 2026, 18(8), 1199; https://doi.org/10.3390/nu18081199 - 10 Apr 2026
Viewed by 693
Abstract
Background and Objectives: This study examined the effects of a novel phytochemical supplement blend on markers of exercise-induced muscle damage. Methods: In a randomised, parallel group design, 24 healthy participants (14 males) consumed 300 mg of a phytochemical blend (calcium fructoborate, turmeric and [...] Read more.
Background and Objectives: This study examined the effects of a novel phytochemical supplement blend on markers of exercise-induced muscle damage. Methods: In a randomised, parallel group design, 24 healthy participants (14 males) consumed 300 mg of a phytochemical blend (calcium fructoborate, turmeric and pomegranate; PB) or inert placebo for 9 days (n = 12 per condition). On day 7, participants performed 150 drop jumps to induce muscle damage. Markers of neuromuscular function, muscle soreness/pain, perceived exhaustion and sleep quality, were measured pre-exercise and 24, 48, and 72 h post-exercise; systemic markers of inflammation, muscle damage, and oxidative stress were measured on these days as well post-exercise and 2.5 h post-exercise. Results: There was an interaction effect for pressure pain threshold in the vastus lateralis (p = 0.041), which was ~21% higher in PB 72 h post-exercise (p = 0.074; ds = 0.767). Perceived sleep quality was greater 72 h post-exercise in PB (p = 0.049; rrb = 0.423) and those in the PB condition reported feeling more recovered and less mentally drained post-exercise (p ≤ 0.043). There were no statistically significant between-condition differences for any markers of neuromuscular function, inflammation, oxidative stress or muscle damage (p > 0.05). Conclusion: In conclusion, a novel PB showed promise for attenuating muscle pain and perceived exhaustion, and improving sleep quality, in the days after muscle damaging exercise. The study protocol was pre-registered on the Open Science Framework Registry (registration number: qgw3a). Full article
(This article belongs to the Section Sports Nutrition)
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21 pages, 7854 KB  
Article
Effect of Dietary Calcium Nitrate Addition on Methane Emission, Nitrogen Excretion, and Ruminal Fermentation Parameters and Microbiota in Liuyang Black Goats
by Mingming Li, Ting Liu, Chen Zheng, Xuan Nan, Jun Wang, Baicong Chen and Hanfang Zeng
Animals 2026, 16(8), 1150; https://doi.org/10.3390/ani16081150 - 10 Apr 2026
Viewed by 417
Abstract
This research explored how dietary supplementation of calcium nitrate influences methane emissions, nitrogen excretion, ruminal fermentation parameters, and microbiota in Liuyang black goats. A total of twelve male goats from this breed were divided into two groups: one serving as a control group [...] Read more.
This research explored how dietary supplementation of calcium nitrate influences methane emissions, nitrogen excretion, ruminal fermentation parameters, and microbiota in Liuyang black goats. A total of twelve male goats from this breed were divided into two groups: one serving as a control group (CON), while the other received a treatment of 3% calcium nitrate (CAL). The research was conducted over a period of 40 days and comprised two separate trial phases. A 10-day adaptation period and a 5-day sampling period (days 11–15) for each stage. Results showed that incorporating calcium nitrate significantly reduced the emissions of methane (CH4) (p < 0.05) and carbon dioxide (CO2) (p < 0.05). Moreover, the use of calcium nitrate modified the trends in ruminal fermentation, resulting in an increase in pH (p < 0.05). Moreover, the ratio of acetate to propionate (A:P) was notably reduced in the CAL group (p < 0.05), indicating a shift toward enhanced production of propionate. At the microbial level, an increased presence of Bacteroidota and Prevotella was observed in the CAL group (p < 0.05). In contrast, the CON group exhibited elevated levels of Firmicutes and Methanobrevibacter (p < 0.05). This finding suggests that calcium nitrate plays a significant role in reducing methane emissions and also affects the fermentation processes in the rumen along with the microbiota of Liuyang black goats. Further research is needed to examine the long-term implications of calcium nitrate supplementation on the health and productivity of these goats. Full article
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30 pages, 2261 KB  
Review
Vitamin K as an Endocrine Modulator: Mechanistic Links to Glucose Metabolism and Beyond
by Wojciech Matuszewski, Mikołaj Madeksza, Michał Szklarz, Aleksandra Rutkiewicz, Joanna Rutkowska and Joanna Maria Harazny
Nutrients 2026, 18(8), 1183; https://doi.org/10.3390/nu18081183 - 9 Apr 2026
Viewed by 743
Abstract
Vitamin K (VK), traditionally recognized for its role in coagulation, is increasingly implicated in extrahepatic processes, including glucose metabolism and calcium regulation. A suboptimal VK status is common in the general population and may limit these functions, yet evidence linking VK to glucose [...] Read more.
Vitamin K (VK), traditionally recognized for its role in coagulation, is increasingly implicated in extrahepatic processes, including glucose metabolism and calcium regulation. A suboptimal VK status is common in the general population and may limit these functions, yet evidence linking VK to glucose metabolism and other endocrine axes remains heterogeneous and incompletely synthesized. This narrative review integrates mechanistic, observational, and interventional evidence to examine the role of VK across the endocrine system, with particular emphasis on glucose metabolism. Mechanistic studies indicate that VK supports pancreatic β-cell function, modulates peripheral insulin sensitivity, and facilitates proper calcium distribution. Observational studies consistently associate a higher VK status with a lower risk of type 2 diabetes, while interventional studies suggest that VK supplementation may improve glucose metabolism, primarily in metabolically impaired populations. In bone and mineral metabolism, VK acts synergistically with calcitriol, with combined supplementation showing more consistent benefits in skeletal outcomes than either vitamin alone. Evidence for VK involvement in other endocrine axes, including reproductive and inflammatory pathways, remains limited and largely mechanistic. Overall, the available evidence supports a context-dependent role for VK in glucose metabolism, influenced by baseline nutritional and metabolic status and outcome selection, as well as a synergistic interaction with calcitriol and parathormone in calcium regulation. Future clinical studies should incorporate baseline VK status stratification, dynamic measures of insulin sensitivity, and adequately powered designs to clarify the therapeutic relevance of VK across endocrine and metabolic outcomes. Full article
(This article belongs to the Section Micronutrients and Human Health)
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