Combined Supplementation Research Articles

Effects of a Novel Dispersible Supplement Containing 2500 IU of Vitamin D and 1000 µg of B12 in Restoring Vitamin D and B12 Insufficiency: A Multicenter, Randomized Controlled Trial

Background/Objectives: Vitamins D and B12 play a crucial role in maintaining bone health, immune function, and neurological integrity. Combined deficiencies in these vitamins can lead to severe health consequences. Current treatment approaches, such as dietary changes and single-vitamin supplementation, often fail to address these deficiencies comprehensively. This study evaluates the effectiveness of concurrent vitamin D and B12 supplementation to correct these insufficiencies. Methods: A prospective, multicenter, randomized controlled trial was conducted in Greece from October 2024 to December 2024. Participants aged 20 to 80 years, with insufficient levels of 25-hydroxyvitamin D (serum < 20 ng/mL) and B12 (serum < 250 ng/L), were eligible for inclusion. Results: A total of 124 patients were randomized into three groups: one receiving vitamins B12 and D in a single supplement (2500 IU + 1000 mcg), one receiving separate doses of each vitamin (2000 IU + 1000 mcg), and a control group receiving no supplementation. The results demonstrated a significant increase in B12 and 25-hydroxyvitamin D levels among the supplemented groups. Particularly, participants in the combined supplementation group showed higher average serum levels of both vitamins. By the end of this study, 37.1% of those in the combined supplement group achieved adequate vitamin levels, compared to 29.4% in the separate supplementation group. Conclusions: In conclusion, combined supplementation may improve patient adherence and compliance, leading to better health outcomes for individuals with combined vitamins D and B12 deficiencies.

Dietary supplementation of blend of organic acids and monoglycerides alleviated diarrhea and systemic inflammation of weaned pigs experimentally infected with enterotoxigenic Escherichia coli F18

BackgroundThe emergence of antibiotic resistant microorganisms associated with conventional swine production practices has increased interest in acid-based compounds having antimicrobial properties and other biological functions as nutritional interventions. Despite the interest in organic acids and monoglycerides, few studies have examined the effects of the combination of these acid-based additives in weaned pigs under disease challenge conditions. Therefore, this study aimed to investigate the effects of dietary supplementation with blend of organic acids and/or medium-chain fatty acid monoglycerides on intestinal health and systemic immunity of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli (ETEC) F18 at 4-week of age.ResultsDietary supplementation of organic acids, monoglycerides, or both organic acids and monoglycerides (combination) reduced (P < 0.05) the diarrhea frequency of ETEC F18-infected pigs throughout the experimental period (d −7 to 21 post-inoculation). This is consistent with the reduced (P < 0.05) proportion of β-hemolytic coliforms in feces observed for the organic acid and combination treatments on d 10 post-inoculation. Supplementation of organic acids, monoglycerides, or combination also reduced (P < 0.05) bacterial translocation in mesenteric lymph nodes on d 21 post-inoculation. Pigs fed with monoglycerides or combination had lower (P < 0.05) white blood cells on d 5 post-inoculation, and pigs fed the combination also had lower (P < 0.05) lymphocytes than pigs in control group. Monoglyceride supplementation increased (P < 0.05) white blood cells and neutrophils compared with control group on d 14 post-inoculation. However, supplementation with organic acid blend, monoglyceride blend, or combination did not affect growth performance in this experiment.ConclusionsSupplementation with monoglycerides or organic acids alone or in combination improves the detrimental effects of ETEC F18 infection in weaned pigs, as indicated by reduced diarrhea, fecal shedding of β-hemolytic coliforms, and bacterial translocation, and thus enhancing disease resistance. Monoglycerides reduced the inflammatory response during peak infection, but their immunomodulatory and possible synergistic effects with organic acids need to be further investigated.

Effects of Quercetin and Citrulline on Nitric Oxide Metabolites and Antioxidant Biomarkers in Trained Cyclists.

Quercetin (QCT) and citrulline (CIT) have been independently associated with improved antioxidant capacity and nitric oxide (NO) production, potentially enhancing cardiovascular function and exercise performance. This study aimed to evaluate the combined and independent effects of QCT and CIT supplementation on NO metabolites and antioxidant biomarkers in 50 trained cyclists undergoing a 20 km cycling time trial (TT). In a randomized, double-blind, placebo-controlled design, forty-two male and eight female trained cyclists were assigned to QCT + CIT, QCT, CIT, or placebo (PL) groups. Supplements were consumed twice daily for 28 days. Biochemical assessments included NO metabolites (nitrate/nitrite), ferric reducing antioxidant power (FRAP), superoxide dismutase (SOD) activity, and antioxidant capacity, measured pre- and post-TT. NO metabolites were significantly elevated post-supplementation (p = 0.03); however, no significant interaction effects were observed for NO metabolites, FRAP, SOD, or antioxidant capacity across the groups (p > 0.05). Post-hoc analyses revealed that QCT significantly reduced FRAP concentrations compared to PL (p = 0.01), while no significant changes in SOD or antioxidant capacity were found across any groups. These findings suggest that combined and independent QCT and CIT supplementation did not significantly improve these biomarkers, suggesting that baseline training adaptations, supplementation timing, and individual variability may influence the efficacy of these compounds in enhancing exercise performance and oxidative stress markers. The ergogenic efficacy of QCT + CIT on antioxidant-related markers remains inconclusive.

Vitamin D3-VDR and vitamin A-RAR affect IL-13 and IFNγ secretion from human CD4+ T cells directly and indirectly via competition for their shared co-receptor RXR.

The effects of vitamin D and vitamin A in immune cells are mediated through the vitamin D receptor (VDR) and retinoic acid receptor (RAR), respectively. These receptors share the retinoid X receptor (RXR) co-factor for transcriptional regulation. We investigated the effects of active vitamin D3 (1,25(OH)2D3) and 9-cis retinoic acid (9cRA) on T helper (TH)1 and TH2 cytokines and transcription factors in primary human blood-derived CD4+ T cells. We aimed to address the discrepancies in this field, particularly regarding the effects of 9cRA and the vitamins in combination. 1,25(OH)2D3 upregulated IL-13 and suppressed IFNγ, while 9cRA had the opposite effects. This was largely independent of a TH1/TH2 phenotype shift. Combined vitamin supplementation produced intermediate cytokine levels, not only through transcriptional regulation by VDR-RXR and RAR-RXR but also through 1,25(OH)2D3 counteracting the effects of 9cRA on solely 9cRA-responsive genes. Similar results were observed in hereditary vitamin D-resistant rickets (HVDRR) patient T cells, where VDR cannot bind to DNA, indicating that RXR binding to either receptor can limit the other's activity. Additionally, we observed downregulated RAR upon 9cRA supplementation and its re-localization out of the nucleus upon 1,25(OH)2D3 supplementation, suggesting a mechanism of indirect regulation by VDR. VDR protein levels were also upregulated upon 9cRA supplementation, suggesting a novel negative feedback mechanism of 9cRA transcriptional activity, whereby 9cRA promotes its own competitor. This study sets the stage for future research into the combined immunomodulatory mechanisms of 1,25(OH)2D3 and 9cRA, involving both direct transcriptional regulation and indirect regulation via RXR competitive binding.

Synergistically effects of n-3 PUFA and B vitamins prevent diabetic cognitive dysfunction through promoting TET2-mediated active DNA demethylation.

Diabetic cognitive dysfunction (DCD) refers to the cognitive impairment observed in individuals with diabetes. Epidemiological studies have suggested that supplementation with n-3 polyunsaturated fatty acid (PUFA) or B vitamins may prevent the development of diabetic complications. Post hoc studies indicate a potential synergistic effect of n-3 PUFA and B vitamins in preventing cognitive impairment. However, the precise effect and underlying mechanism of this combination on DCD remain unclear. In case-control study, we compared fatty acid composition of erythrocyte membrane and serum homocysteine levels between diabetic individuals with and without DCD. We found that insufficient levels of n-3 PUFA, along with elevated serum homocysteine, significantly increase the risk of developing DCD. Treatment with a combination of fish oil, folate, and vitamin B12 improved cognitive impairment and aberrant neuronal morphology in streptozotocin-induced DCD mice. Folic acid and vitamin B12 enhanced the efficiency of exogenous docosahexaenoic acid (DHA) transportation to the brain by preventing the accumulation of homocysteine and S-adenosylhomocysteine, thereby inhibiting neuronal apoptosis in diabetic brains. Furthermore, folic acid and vitamin B12 supplementation can provide sufficient 5-methylcytosine for diabetic brains by promoting DNA methylation, while increased DHA levels maintain TET-mediated active DNA demethylation in diabetic brains through enhancing TET2 function. Overall, our study provides novel insights into molecular mechanisms underlying the synergistic preventive effects of the combined supplementation with fish oil, folic acid and vitamin B12 on DCD, suggests that combining n-3 PUFA and B vitamins could be a promising strategy for preventing DCD among individuals with diabetes.

Effect of Dietary Supplementation with Lutein, Zeaxanthin, and Elderberries on Dry Eye Disease (DED) and Immunity: A Randomized Controlled Trial.

Dry eye disease (DED) significantly impairs quality of life, affecting physical, social, and psychological well-being, as well as reducing workplace productivity. While lutein and zeaxanthin supplements have been shown to improve ocular health, existing research often overlooks the efficacy of lower dosages and shorter durations of supplementation. This study investigated the effects of combined supplementation with lutein, zeaxanthin, and elderberries in 110 voluntary participants through a randomized controlled trial. Participants took 6 mg of lutein and 1 mg of zeaxanthin, along with 100 mg elderberry extract once daily for a duration of 20 days. Ocular health was assessed using the Ocular Surface Disease Index (OSDI), while immune status was evaluated with the Immune Status Questionnaire (ISQ). Results showed that combined supplementation significantly (p < 0.05) reduced the OSDI scores in the intervention group from 38.15 ± 11.14 to 18.26 ± 5.57, reflecting a 52.2% reduction. A similar trend was observed with the Visual Analog Scale (VAS), indicating significant (p < 0.05) improvement from 5.31 ± 1.62 to 6.73 ± 1.74, equivalent to a 26.7% improvement. Although the intervention group showed a 15.9% improvement in ISQ scores by the study's end, this was not significantly different from the placebo group, suggesting that higher dosages or longer durations may be needed to observe a meaningful effect. Additionally, findings from the Food Frequency Questionnaire revealed that the average dietary intake of lutein and zeaxanthin among participants was only 663.49 µg, equating to just 5.5% of the suggested optimal daily intake. This low consumption is concerning, as it is inversely correlated with the risk of ocular diseases. Collectively, these findings support the use of combined supplementation as an adjuvant approach to improving ocular health.

International Symposium on Ruminant Physiology: One-carbon metabolism in beef cattle throughout the production cycle.

One-carbon metabolism (OCM) is a series of connected pathways involving the methionine-folate cycles, transsulfuration, polyamine synthesis, nucleotide synthesis, free-radical scavenging, and energy metabolism. These pathways functionally depend upon amino acids (methionine, glycine, and serine), vitamins (folate, B2, B6, and B12), and minerals (sulfur, cobalt, and zinc). Growing bodies of research indicate that in beef cattle, physiological stage, nutritional plane, diet, species (Bos taurus vs. indicus), rumen protected vs. not, individual vs. combination supplementation and method of delivery all affect the efficacy of one-carbon metabolite supplementation. Infusion studies showed that supplementing methionine to growing steers improved N retention and altered hepatic activity of methionine synthase; however, only supplementing methionine without folate decreased folate concentrations in circulation. When heifers were supplemented with methionine, choline, folate, and B12 for the first 63 d of gestation, metabolomic analysis revealed increasing OCM analytes to the heifer, but a buffering effect to the fetus with minimal changes seen in hepatic metabolite abundance. Methionine supplementation to heifers during the periconceptual period increased circulating methionine but shifted fetal hepatic metabolism toward the transsulfuration pathway. Periconceptual methionine supplementation to cows increased gain and total-tract digestibility in calves post-weaning. In vitro supplementation of choline to beef cattle embryos results in calves of increased birth and weaning weight. Overall, these data demonstrate that OCM is altered in those cattle receiving one-carbon metabolites, and that a metabolic programming response is elicited in offspring receiving supplements in vitro or during early gestation. Research should be considered to maximize efficiency of beef cattle production at all stages by identifying limiting metabolites or enzymes to maximize efficiency of OCM in beef cattle, as well as to understand the concerted effects of multiple one-carbon metabolites to balance the stoichiometry of the pathway.

Safety and Efficacy of Loading Doses of Vitamin D: Recommendations for Effective Repletion.

Background/Objectives: Epidemiological data on vitamin D status revealed that, despite various dosage and durations of supplementation, the effectiveness often fails to achieve optimal outcomes. The need for higher doses than previously recommended was suggested, but several modifying factors should be considered, including the level of deficiency, and BMI. The objectives of this post hoc evaluation are to characterize treatment effectiveness based on the applied dose, duration and BMI; and to assess the safety aspects associated with rapid repletion of vitamin D. Methods: Vitamin D deficient subjects selected in the post-hoc analysis: seventy patients included from a combined loading-maintenance supplementation (300,000 IU followed by 60,000 IU) protocol and 62 deficient subjects who received a low-dose maintenance (1000 IU/day) therapy. The risk of overload and the incidence of hypercalciuria and hypercalcemia resulting from loading or post-loading maintenance were investigated. Results: The moderate-fast-loading schedule of 60,000 IU per week for 5 weeks, effectively achieves the target in 25(OH)D levels over 30 ng/mL for all deficient subjects, regardless of their BMI. Slower loading with lower weekly doses confirms the safety of supplementation, but the effectiveness is dependent on the subjects' BMI; overweight and obese patients require higher doses to reach the same vitamin D levels. No difference in safety parameters observed compared to low-dose therapies. Conclusions: The loading treatment involving a total dose of 300,000 IU administered over 5 or 10 weeks is effective for repletion, does not lead to 25(OH)D overload, and poses no additional risks of hypercalcemia or hypercalciuria. Furthermore, there are no safety concerns regarding changes in bone resorption markers. A combination of the loading treatment with a subsequent maintenance dose of 2000 IU daily is adequate to achieve the target vitamin D levels.

Effects of Zinc and Selenium Coadministration on Semen Parameters and Hormonal Profiles of Male Wistar Rats.

Zinc (Zn) and Selenium (Se) are essential dietary micronutrients or trace elements not produced by the body but required in small amounts for the maintenance of mammalian and human male fertility. The over supplementation of individual and combination of micronutrients in the sick and healthy populations could be the leading cause of prevalence of male factor infertility in the world. This study was designed to evaluate the possible effects of the graded independent doses of oral supplementation of Zn, Se and their co-administration on male reproductive hormones, sperm parameters and histological architecture of male wistar rats. Fifty (n=50) adult male wistar rats weighing between 150g-250g and randomly grouped into ten groups of five rats each were used for this research that lasted for six weeks. Group I, the control group, received normal rat chow and water ad libitum. The nine test groups (II-X), received graded doses of zinc sulphate ZnSO4 (II-IV: 50mg/kg,150mg/kg,300mg/kg), selenium dioxide SnO2 (V-VII; 100mg/kg,200mg/kg,400mg/kg), co-administration of ZnSO4 and SnO2 (VIII-X:50mg/kg ZnSO4+100mg/kg SeO2, 150mg/kg ZnS04+200mg/kg SeO2, 300mg/kg ZnSO4+400mg/kg SnO2). Serum testosterone (T), follicular stimulating hormone (FSH) and luteinising hormone (LH) was estimated using ELISA Hormone Test kits radio-immuno-assay technique. Semen parameters was analyzed for sperm count, motility, and morphology using the grading system as described by WHO (1999). Testicular histological architecture was established using animal tissue processing technique. Data obtained were analyzed using one-way Analysis of Variance (ANOVA) from SPSS version 22. Statistical significance was placed at P&lt;0.05 and data expressed as mean ± standard error of the mean (SEM).Results obtained from the study showed significant increase in testosterone levels in all the Zn test groups compared to the control and significant decrease in the levels of FSH and LH in higher doses of Zn and Se supplements when compared to the control at p&lt;0.05. There was significant reduction in sperm motility, count but increase sluggishness in Zn and co-administration of Zn and Se of all the test groups compared to the control. Photomicrograph of the testes showed tissue inflammation, germinal cells destruction and azospermia. This study showed that chronic single or combined over supplementation of the micronutrients zinc and selenium could cause male factor infertility in wistar rats.

RESPONSE OF SUMMER TOMATO WITH EXOGENOUS APPLICATION OF SALICYLIC ACID AND CALCIUM

A pot experiment was carried out at Horticulture Farm, Sher-e-Bangla Agricultural University, Dhaka May to September, 2016 to evaluate the performance of summer tomato production with the supplementation of salicylic acid (SA) and calcium (Ca). The experiment consisted of two factor. Four different levels of SA (S1 = 0 mM SA, S2 = 0.3 mM SA, S3 = 0.6 mM SA, S4 = 0.8 mM SA) and three levels of Ca (C1 = 0 mM Ca, C2 = 4 mM Ca, C3 = 8 mM Ca). Both morphological traits and yield of summer tomatoes were significantly affected by the application of SA and Ca. Exogenous application of SA and Ca influenced the growth including plant height, leaf number, branch number, yield contributing characters and yield. The combined supplementation of 0.8 mM SA and 8 mM Ca resulted the highest yields (0.8 kg/plant) where lowest one (0.3 kg/plant) was recorded from the control treatment (without SA and Ca). These results suggest that SA and Ca can improve the growth and yield of tomatoes during the summer season by alleviating the detrimental effects of extreme heat in the summer.

Efficacy of a combined exercise and nutrition intervention study for outpatients with possible sarcopenia in community-based primary care clinics (ENdSarC): study protocol for a multicenter single-blinded randomized controlled trial

BackgroundSarcopenia is a geriatric disease characterized by loss of muscle mass and strength. Although combined exercise and nutrition intervention are known to be effective for sarcopenia, clinical trials involving outpatients with sarcopenia in primary care are scarce. We describe a protocol for a trial to examine the effects of a 12-week combined exercise and nutrition intervention in Korean older adults with possible sarcopenia in community-based primary care.MethodsThis multicenter, randomized, controlled trial will include 94 community-dwelling older outpatients aged 65–85 years with possible sarcopenia (47 participants in the intervention and control groups each). Resistance exercises, which incorporate concentric and eccentric exercises, will consist of an introductory phase (3 weeks: twice-weekly supervised exercise sessions and once-weekly home exercises; contraction exercises), an expanded phase (3 weeks: twice-weekly supervised exercise sessions and once-weekly home exercises; eccentric exercises), and a maintenance phase (6 weeks: once-weekly supervised exercise sessions and twice-weekly home exercises; power/eccentric exercises). Nutritional supplementation will be provided according to the nutritional status of the participants using a Mini-Nutritional Assessment. Participants will be assessed at baseline, 12 and 24 weeks, and the primary outcome will be the 5-times chair stand test results.DiscussionTo the best of our knowledge, this will be the first clinical trial to evaluate the efficacy of a combined exercise and nutritional supplementation intervention in older outpatients with possible sarcopenia in community-based primary care clinics. These findings will provide new insights to clinicians regarding the long-term usability for doctors and outpatients with possible sarcopenia in community-based primary care.Trial registrationThis trial was prospectively registered at ClinicalTrials.gov on September 16, 2023 (registration number: NCT06049914).

Absorption, anti-inflammatory, antioxidant, and cardioprotective impacts of a novel fasting mimetic containing spermidine, nicotinamide, palmitoylethanolamide, and oleoylethanolamide: A pilot dose-escalation study in healthy young adult men

This pilot dose-escalation study evaluated the absorption and metabolism of a novel fasting mimetic formulation containing spermidine, nicotinamide, palmitoylethanolamide (PEA), and oleoylethanolamide (OEA) taken as oral supplements in young adults. Five healthy men consumed a standardized breakfast, followed by control (wheat flour) or low, medium, or high doses of supplements containing spermidine, nicotinamide, PEA, and OEA 2 hours later. Blood was drawn at 0, 1, 2, and 4 hours after the supplement (2, 3, 4, and 6 hours postprandial). Plasma concentrations of spermidine, 1-methylnicotinamide, PEA and OEA were quantified by liquid chromatography-mass spectrometry. The secretion of tumor necrosis factor alpha and production of reactive oxygen species by stimulated macrophages incubated with plasma, and cholesterol efflux capacity of plasma were analyzed. Plasma 1-methylnicotinamide, PEA, and OEA concentrations increased after supplement intake (P < .05). Spermidine concentrations decreased in the control arm (P < .05) but not the supplement arms. Net incremental area under the curve for tumor necrosis factor alpha and reactive oxygen species in stimulated macrophages decreased when incubated with plasma following supplement intake (P < .05). Intake of the combined supplements showed they were bioavailable and increased in plasma in a dose-dependent manner and provide preliminary data showing enhanced plasma anti-inflammatory and antioxidant functions.This trial was registered at clinicaltrials.gov (NCT05017428).

Glutathione and Selenium Supplementation Attenuates Liver Injury in Diethylnitrosamine-Induced Hepatocarcinogenic Mice by Enhancing Glutathione-Related Antioxidant Capacities.

Excess oxidative stress and inadequate antioxidant capacities are critical features in the development of hepatocellular carcinoma. This study aimed to determine whether supplementation with glutathione (GSH) and/or selenium (Se), as antioxidants, attenuates diethylnitrosamine (DEN)-induced hepatocarcinogenesis in mice. C57BL/6J male mice were randomly assigned to control, DEN, DEN + GSH, DEN + Se, and DEN + GSH + Se groups for 20 weeks. Daily supplementation with GSH and/or Se commenced in the first experimental week and continued throughout the study. DEN was administered in weeks 2-9 and 16-19 of the experimental period. DEN administration induced significant pathological alterations of hepatic foci, evidenced by elevated levels of liver function, accompanied by high malondialdehyde (MDA) levels; low GSH levels; and glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S-transferase (GST) activities. Supplementation with GSH and Se significantly ameliorated liver pathological changes, reducing liver function and MDA levels while increasing GSH levels and GPx, GR, and GST activities. Notably, combined supplementation with GSH and Se more effectively increased the GSH/glutathione disulfide ratio and GPx activity than individual supplementation. Supplementation with GSH and Se attenuated liver injury in DEN-induced hepatocarcinogenic mice by enhancing GSH and its related antioxidant capacities, thereby mitigating oxidative damage.

The impact of tangeretin combined with whey protein on exercise-induced bronchoconstriction in professional athletes: a placebo-controlled trial.

Exercise-induced bronchoconstriction (EIB) is highly prevalent in athletes. The objective of this study was to assess the therapeutic efficacy of daily tangeretin combined with whey protein supplementation over a period of 4weeks in professional athletes with EIB. Using a placebo-controlled, double-blind, paired, randomized trial design, a cohort of 30 professional athletes with EIB, consisting of 14 females and 16 males, was divided into two groups: the tangeretin combined with whey protein intervention group (TIG), and the placebo control group (PCG). Both the TIG and PCG underwent exercise challenge tests (ECT) and VO2max tests before (ECT1, V1) and after (ECT2, V2) the intervention. Blood (eosinophils, neutrophils, and basophils) and serum (interleukin-5, IL-5; interleukin-8, IL-8; Clara cell secretory protein-16, CC16; immunoglobulin E, IgE) levels were measured early in the morning of ECT1 and ECT2, respectively. Lung function was assessed immediately before and post-ECT immediately. Tangeretin combined with whey protein use for 4 weeks attenuated the decrease in forced expiratory volume in 1 s (FEV1) post trials (∆FEV1(ECT1-ECT2): mean (SD) TIG -7.51(6.9)% vs. PCG -2.33(11.49)%, p = 0.013). Tangeretin also substantially attenuated IL-5 concentration (∆IL-5(T1-T5): Tangeretin -19.4% vs Placebo + 8.37%, p = 0.022); IL-8 concentration (∆IL-8(T1-T5): Tangeretin -17.28% vs Placebo + 6.1%, p = 0.012); CC16 concentration (∆CC16(T1-T5): Tangeretin -11.77% vs Placebo + 24.19%); and IgE concentration in the serum (∆IgE(T1-T5): Tangeretin -24.1% vs Placebo -3.9%), and significantly decreased neutrophil count (∆N(T1-T5): Tangeretin -11.34% vs Placebo + 0.3%) and eosinophil count in blood (∆N(T1-T5): Tangeretin -38.5% vs Placebo + 4.35%). Compared with V1, VO2max (p = 0.042) and TLim (p = 0.05) of V2 were significantly increased in the TIG, and there was no significant change in the PCG. Meanwhile, six athletes in the TIG and 0 athletes in the PCG became EIB-negative at ECT2; the overall negative conversion rate of EIB was 40.00% in TCG. Additionally, the number of cough symptoms decreased from 9 to 3 and dyspnea from 4 to 2 in the TIG. After high-intensity exercise, athletes with EIB achieved significant improvements in lung function and blood inflammatory factors by combining tangeretin and whey protein supplementation. EIB athletes also showed longer exercise endurance and VO2max at 4 weeks after TI. In addition, some patient symptoms disappeared after combination supplementation. The effect of this treatment on professional athletes with EIB was beneficial.

The Effect of Selenium, Zinc, and their Combined Supplementation on Cardiometabolic Biomarkers-comparing their Effects in the Energy Restriction and High-fat Diet Methods in Obese Rats.

The fat distribution in the body determines the risk of cardiometabolic problems such as heart disease and diabetes. Some dietary supplements, such as selenium and zinc, possess lipolytic and anti-angiogenic functions, which may be a useful strategy in reducing the risk of cardiometabolic complications. This study evaluated the effect of zinc (Zn), selenium (Se), and their combined supplementation on cardiometabolic risk factors in male Wistar rats in two nutritional models, including caloric restriction (CR) and high-fat diet (HFD). The 48 male Wistar rats were divided into three diet groups (HFD and CR and normal diet (ND)). The HFD group was subdivided into four groups (N=8 rats in each group) that received (HFD+Se), (HFD+Zn), (HFD+Zn+Se), and HFD alone as the control group, respectively. After 8 weeks of intervention, biochemical tests were performed on serum levels, including measurement of lipid profile (triglyceride, Cholesterol, LDL and HDL) and glycemic indices (fasting blood sugar, insulin and insulin sensitivity markers). The results showed that supplementation significantly improved the lipid profile (P <0.001). A comparison of glucose homeostasis indices in the study groups also showed a significant difference. The serum level of glucose was higher in the HFD group than in the intervention groups (P <0.001). Also, the rate of improvement of lipid profile and glycemic indexes in the group receiving the combination of two supplements showed a better trend than those receiving zinc and selenium alone. However, the values were statistically significant only for glucose homeostasis indices (P <0.001). Although obesity is a multifactorial condition, controlling other risk factors, zinc and selenium and their combined supplementation can lead to promising solutions for the treatment of obesity-induced glucose and lipid homeostasis disorders.

Synergistic Benefits of Dietary Silymarin and Selenium on Growth, Immune Functions, Antioxidants, and Gut/Liver Health of Thinlip Mullet (Liza ramada) Juveniles

Abstract This study investigates the synergistic impact of silymarin (SI) levels combined with inorganic selenium (sodium selenite: Se) on growth, feed utilization, biochemical parameters, antioxidants, innate immunity, intestinal and liver histology, and gene expression of thinlip mullet (Liza ramada) juveniles. The experimental design involved thinlip mullets initially weighing 3.5±0.13 g, distributed in a completely randomized design with 30 fish per hapa (0.5 × 0.5 × 1 m), and conducted in triplicate over 60 days. Seven experimental diets were employed, including a control (without SI and Se supplementation), a negative control (with only Se supplementation), and four treatments with varying levels of silymarin (250, 450, 650, 850 mg/kg) alongside selenium (0.5 mg/kg diet). The growth performance results highlighted significant enhancements in final body weight, weight gain, and specific growth rate, particularly in the SI 850 mg/kg + Se treatment. Survival rates, feed intake, and feed conversion ratios showed positive trends across the SI-Se supplemented groups. Biochemical profiles of serum exhibited that the control diet induced elevated concentrations of glucose, cholesterol, alanine aminotransferase, aspartate aminotransferase, and urea, while Se or SI supplementation significantly mitigated these levels, with the lowest concentrations observed in the SI-Se supplemented groups. Moreover, SI supplementation increased serum protein content. Antioxidant enzyme activities, represented by superoxide dismutase (SOD), catalase (CAT), and catalase (GPx), demonstrated notable improvements in the SI-Se fortified groups, with significantly elevated GPx activity compared to the Se-supplemented and control groups. Immune system responses, including lysozyme, bactericidal, nitro-blue tetrazolium (NBT%), and serum alternative complement pathway (ACH50) activities, were highest in the SI-Se augmented groups. SI and Se in L. ramada reduce liver pro-inflammatory gene expression (IL-1 β, hepcidin) vs. control group. Histological examinations of the intestine and liver depicted structural enhancements, especially at moderate and high levels of SI with Se supplementation. The results indicate improved intestinal villi morphology and hepatic architecture, supporting the positive influence of dietary treatments on the health of thinlip mullet juveniles. In conclusion, the combined supplementation of SI at 850 mg/kg diet and Se at 0.5 mg/kg diet positively influenced the growth, biochemical profiles, antioxidant status, immune responses, gene expression, and histological integrity of thinlip mullet juveniles, providing valuable insights for optimizing aquafeed formulations.

Investigating The Effect of Combined Nutritional Supplementation in the Management of Autism Spectrum Disorder and the Progression of Social and Communication Skills: A Nonrandomized Interventional Study

In recent years, there has been an increase in the use of nutritional supplementation as a method of treating autism spectrum disorder (ASD). The aim of our study is to investigate the effect of using combined supplementation on treating the symptoms of Autism. In this study, we conducted a nonrandomized, nonblinded trial where we followed up with patients with ASD who were diagnosed through the CARS2-ST scale and evaluated through the Vineland-3 scale within two visits. The patients were divided into control and intervention groups where training only and supplementation with training were used, respectively. Results: The study included a total of 115 participants, out of which 95 (82.6%) were assigned to the intervention group and 20 (17.4%) were assigned to the control group. Significant improvement was observed in the intervention group compared to the control group. It was observed that the CARS2 score for the intervention group was reduced by a mean of 16.1 points compared to only 4.45 mean points reduced in the control group. On the other hand, the Vineland-III scale was increased by a mean of 350.23 points in the intervention group, while the control group had an increase of only 41.3 points. Conclusion: Our study concludes that the use of combined supplementation along with behavioral training greatly reduces autism symptoms compared to training alone. Thus, our study can recommend the use of combined supplementation as a form of treatment for Autism. Further studies with randomization and blinding will be helpful in confirming the findings of our study.

Effects of passion fruit peel (Passiflora edulis) pectin and red yeast (Sporodiobolus pararoseus) cells on growth, immunity, intestinal morphology, gene expression, and gut microbiota in Nile tilapia (Oreochromis niloticus)

This study explores the effects of dietary supplementation with passion fruit peel pectin (Passiflora edulis) and red yeast cell walls (Sporidiobolus pararoseus) on growth performance, immunity, intestinal morphology, gene expression, and gut microbiota of Nile tilapia (Oreochromis niloticus). Nile tilapia with an initial body weight of approximately 15 ± 0.06 g were fed four isonitrogenous (29.09–29.94%), isolipidic (3.01–4.28%), and isoenergetic (4119–4214 Cal/g) diets containing 0 g kg−1 pectin or red yeast cell walls (T1 - Control), 10 g kg−1 pectin (T2), 10 g kg−1 red yeast (T3), and a combination of 10 g kg−1 pectin and 10 g kg−1 red yeast (T4) for 8 weeks. Growth rates and immune responses were assessed at 4 and 8 weeks, while histology, relative immune and antioxidant gene expression, and gut microbiota analysis were conducted after 8 weeks of feeding. The results showed that the combined supplementation (T4) significantly enhanced growth performance metrics, including final weight, weight gain, specific growth rate, and feed conversion ratio, particularly by week 8, compared to T1, T2, and T3 (P < 0.05). Immunological assessments revealed increased lysozyme and peroxidase activities in both skin mucus and serum, with the T4 group showing the most pronounced improvements. Additionally, antioxidant and immune-related gene expression, including glutathione peroxidase (GPX), glutathione reductase (GSR), and interleukin-1 (IL1), were upregulated in the gut, while intestinal morphology exhibited improved villus height and width. Gut microbiota analysis indicated increased alpha and beta diversity, with a notable rise in beneficial phyla such as Actinobacteriota and Firmicutes in the supplemented groups. These findings suggest that the combined use of pectin and red yeast cell walls as prebiotics in aquaculture can enhance the health and growth of Nile tilapia, offering a promising alternative to traditional practices. Further research is needed to determine optimal dosages for maximizing these benefits.

Effects of Carica Papaya Leaves and Fermented Fish Waste on the Production Performance of Broiler Chickens

Introduction: Poultry feed management requires novel dietary additives to enhance chicken performance and manage feed availability. The present study investigated the effects of dietary supplementation of Carica papaya leaf meal (CPL) and fermented fish waste (FFW) in water on different growth indicators, such as body weight, carcass yield, feed intake, and conversion rate of Bovans brown male broiler chickens. Materials and methods: A randomized complete block design with a 2x4 factorial arrangement was used to study the effect of the combination supplementation of the 10% CPL with FW at 0%, 5%, 10%, and 15%. Both additives combination treatment was replicated four times with five broiler chicks per replication, a total of 160-day-old male Bovans brown broilers with 39-40g body weight were used in the current study. The duration of the study was 20 days (from the 16th to 35th days of age). Results: Studies have revealed that CPL negatively impacted the final body weight, whereas FFW positively influenced feed intake, final weight, and weight gain. Moreover, 15% of FFW supplementation resulted in noteworthy increases in drumstick and thigh weights compared to the control group, with significant interactions identified between CPL and FFW for breast, wing, and back weights. However, neither CPL nor FFW had an impact on the weights of edible visceral organs. The palatability of meat quality assessment showed no significant difference among the treatments. Adding CPL and FFW increased weights and profits, with the best rate of intake observed in broilers given the standard diet plus 15% of FFW supplementation. Conclusion: These results indicated that using CPL and FFW at 10% and 15%, respectively was a financially feasible approach to improve broiler development, carcass quality, and profitability while capitalizing on nutritional advantages and reducing environmental waste.

Impact of spinach thylakoid extract-induced 12-week high-intensity functional training on specific adipokines in obese males.

Obesity presents multifarious etiopathologies with its management being a global challenge. This article presents the first ever report on the impact of spinach thylakoid extract-induced high-intensity functional training (HIFT) on obesity management via regulating the levels of novel adipokine, C1q/TNF-related Protein-12 (CTRP-12), furin, and Krüppel-like factor 15 (KLF-15). Sixty-eight obese male subjects were randomly divided into four groups: control group (CG), supplement group (SG), training group (TG), and the combined training and supplement group (TSG). After initial assessments of all groups, the training group commenced a twelve-week HIFT using the CrossFit program (comprising of three training sessions per week, each lasting 30 min). Eligible candidates were randomly assigned to either receive thylakoid-rich spinach extract (5 g per day) or a matching placebo (5 g per day of corn starch, 30 min before lunch) for a total duration of 12 weeks. All required data and investigations were collected at 48 h pre- and post-training. The results indicated a substantial correlation between exercise and the time of KLF-15, furin, and CTRP-12 demonstrating effect sizes of 0.3, 0.7, and 0.6, respectively. Additionally, the training and supplementation group (TSG) exhibited a substantial decrease in low-density lipoprotein (LDL), total cholesterol (TC), and triglyceride (TG) levels (p < 0.0001). Concurrently, there was a significant increase in high-density lipoprotein-cholesterol (HDL-C) levels (p = 0.0001). Furthermore, a notable difference between the groups emerged in HDL, LDL, TC, and TG levels, supported by effect sizes of 0.73, 0.86, 0.96, and 0.89, respectively (p < 0.05). The study offered novel insights into the management of obesity using supplements induced by spinach-derived thylakoid extract during a 12-week HIFT program. The proposed combination intervention may reverse obesity-induced insulin resistance and metabolic dysfunctions by positive regulation of CTRP-12/adipolin and KLF15 and simultaneous suppression of furin levels.