Long-term Supplementation Research Articles

Thermogenic Adipocytes Promote M2 Macrophage Polarization through CNNM4-Mediated Mg Secretion.

M2 macrophages promote adipose tissue thermogenesis which dissipates energy in the form of heat to combat obesity. However, the regulation of M2 macrophages by thermogenic adipocytes is unclear. Here, it is identified magnesium (Mg) as a thermogenic adipocyte-secreted factor to promote M2 macrophage polarization. Mg transporter Cyclin and CBS domain divalent metal cation transport mediator 4 (CNNM4) induced by ADRB3-PKA-CREB signaling in thermogenic adipocytes during cold exposure mediates Mg efflux and Mg in turn binds to the DFG motif in mTOR to facilitate mTORC2 activation and M2 polarization in macrophages. In obesity, downregulation of CNNM4 expression inhibits Mg secretion from thermogenic adipocytes, which leads to decreased M2 macrophage polarization and thermogenesis. As a result, CNNM4 overexpression in adipocytes or Mg supplementation in adipose tissue ameliorates obesity by promoting thermogenesis. Importantly, an Mg wire implantation (AMI) approach is introduced to achieve adipose tissue-specific long-term Mg supplement. AMI promotes M2 macrophage polarization and thermogenesis and ameliorates obesity in mice. Taken together, a reciprocal regulation of thermogenic adipocytes and M2 macrophages important for thermogenesis is identified, and AMI is offered as a promising strategy against obesity.

Citrus supplementation in subjective cognitive decline: results of a 36-week, randomized, placebo-controlled trial

BackgroundDeveloping interventions for older adults with subjective cognitive decline (SCD) has the potential to prevent dementia in this at-risk group. Preclinical models indicate that Citrus-derived phytochemicals could benefit cognition and inflammatory processes, but results from clinical trials are still preliminary. The aim of this study is to determine the effects of long-term supplementation with Citrus peel extract on cognitive performance and inflammation in individuals with SCD.MethodsEighty participants were randomly assigned to active treatment (400 mg of Citrus peel extract containing 3.0 mg of naringenin and 0.1 mg of auraptene) or placebo at 1:1 ratio for 36 weeks. The primary endpoint was the change in the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) total score across the 36-week trial period. Other cognitive outcomes included tests and scales evaluating verbal memory, attention, executive and visuospatial functions, and memory concerns. The secondary endpoint was the change of interleukin-8 (IL-8) levels over the 36-week trial period in a subsample of 60 consecutive participants. An Intention-to-treat approach with generalized linear mixed models was used for data analysis.ResultsThe RBANS total score showed significant improvement in both Citrus peel extract and placebo groups at 36 weeks (p for time < .001, d = 0.36, p time x treatment = .910). Significant time effects were also found in cognitive domains of short- and long-term verbal memory (p < .001) and scales of subjective memory (p < .01), with no significant time x treatment interaction. The largest effect sizes were observed in verbal memory in the placebo group (d = 0.69 in short-term, and d = 0.78 in long-term verbal memory). Increased IL-8 levels were found at 36-week follow-up in both Citrus peel extract and placebo groups (p for time = .010, d = 0.21, p time x treatment = .772). Adverse events were balanced between groups.ConclusionsIn this randomized clinical trial, long-term Citrus peel extract supplementation did not show cognitive benefits over placebo in participants with SCD, possibly due to high placebo response. These findings might have specific implications for designing future nutraceutical trials in individuals experiencing SCD.Trial registrationThe trial has been registered at the United States National Library of Medicine at the National Institutes of Health Registry of Clinical Trials under the code NCT04744922 on February 9th, 2021 (https://www.clinicaltrials.gov/ct2/show/NCT04744922).

Blackcurrant Supplementation and Habitual Physical Activity Enhance Functional Capacity and Quality of Life in Previously Sedentary Older Women.

Loss of functional capacity negatively impacts the quality of life in older adults. This single-site randomized controlled study examined whether short- (one week) and long-term (three months) blackcurrant supplementations combined with habitual physical activity would improve functional capacity and quality of life in previously sedentary older women. Thirty sedentary, healthy older women were randomly assigned to receive blackcurrant supplementation (400 mg of blackcurrant extract) (Polyphenol group, n=15; 74.2±10.0 years) or no supplementation (capsules of 400 mg of corn flour) (Placebo group, n=15; 72.8±8.7 years). Each group consumed two capsules daily for one week and three months, with a washout period of one week between the two phases. The polyphenol group was required to increase habitual activity levels. Participants underwent a functional capacity assessment consisting of a sixminute walk, sit-to-stand test, Berg balance scale measurement and quality of life evaluation with the Scale of Life Satisfaction Index (LSI) at the start, after one week, and after three months of supplementation. Walking distance and time to sit-to-stand improved by 2.5% (p=0.005) and 7.5% (p=0.005), respectively, after one week in the polyphenol group. After 3 months, walking distance in the six-minute walk test increased by 12.3% (p=0.001) while the time to sit-to-stand decreased by 16% (p=0.002) in the polyphenol compared to placebo group, respectively. No differences in Berg balance scale were observed. Quality of life, indexed by LSI, improved by 39% (p=0.001) in the polyphenol compared to the placebo group. Blackcurrant supplementation, combined with habitual activity, may enhance functional capacity and quality of life in older women, offering a potential strategy to maintain independence. However, future studies should address longer durations to validate these findings.

Effect of vitamin D supplementation on cardiovascular outcomes: an updated meta-analysis of RCTs.

To evaluate the effect of vitamin D supplementation on cardiovascular outcomes. After searching different databases, we retrieved and included randomized controlled trials on long-term supplementation of vitamin D (≥1-year intervention) and reporting cardiovascular outcomes. We calculated risk ratio (RR) with 95% confidence intervals (CI) for dichotomous outcomes. Compared to the control group, the vitamin D group was not associated with a statistically significant decrease in the incidence of major adverse cardiovascular events (MACE) [risk ratio=0.99; 95% CI: 0.94-1.03]. We found no difference between the vitamin D group and the control group for the outcomes of incidences of myocardial infarction, heart failure, coronary revascularization, cardiovascular death, and all-cause mortality. The heterogeneity was low for all outcomes. According to our meta-analysis, vitamin D supplementation did not reduce major adverse cardiovascular events, other cardiovascular parameters, and all-cause mortality.

Short-Term Zinc Supplementation Stimulates Visceral Adipose Catabolism and Inflammation in Mice.

Zinc (Zn), a fundamental trace element in human biology, exhibits pivotal roles in sustaining vital physiological processes and regulating metabolic homeostasis. Insufficient zinc intake has been linked to deleterious consequences on growth, reproductive functions, metabolic activities, and immune responses in both humans and animals. Oral zinc supplementation is usually performed to meet zinc requirement. Previous studies have shown that long-term supplementation of zinc in mice impaired AKT signaling and induced adipocyte hypertrophy in visceral adipose tissue. The presented study was conducted to investigate the role and mechanism of short-term zinc supplementation on lipids metabolism. Zinc sulfate was supplemented in the drinking water of C57/BL6J male mice at 30 ppm or 90 ppm for one week. Water consumption, food intake, and body weight were analyzed, adipose tissue and serum profile of metabolites were investigated, and the key genes related to lipid metabolism were analyzed. Short-term zinc supplementation decreased visceral adipose tissue weight and adipocyte size compared to the control group, but no difference was observed in food intake, water consumption, and body weight between the two groups. Further studies revealed that short-term zinc supplementation significantly increased the serum insulin level while decreasing the serum NEFA content. In addition, zinc supplementation increased the expression of Atgl and Hsl in the visceral adipose tissue compared with the control mice. Furthermore, the phosphorylation level of HSL and protein level of PPARg in the epididymal adipose tissue increased by zinc supplementation compared with the control mice. In comparison, the protein level of FASN was down-regulated by short-term zinc supplementation in the epididymal adipose tissue, although the expression of lipogenic genes was not changed. The expression of F4/80 and Tnfa were increased in zinc-supplemented adipose tissue as compared with the control group. Our findings suggest that short-term zinc supplementation might reduce fat deposition by enhancing lipolysis in mice. Future studies could focus on the effect of intermittent zinc supplementation on fat reduction in both animal models and humans.

Updated Meta-analysis Reveals Limited Efficacy of Vitamin D Supplementation in Chronic Low Back Pain.

Chronic low back pain (CLBP) significantly reduces quality of life and increases reliance on healthcare resources. Despite many individuals opting for vitamin D supplementation to alleviate CLBP, its efficacy remains debatable. This meta-analysis aimed to evaluate the potential benefits of vitamin D supplementation in treating this condition. Adhering to PRISMA guidelines, we systematically reviewed the effectiveness of vitamin D supplementation in adults with CLBP, focusing exclusively on randomized controlled trials (RCTs). A comprehensive literature search was conducted up to May 2024 across multiple databases, including PubMed, Scopus, Cochrane Library, and Web of Science. Ten RCTs meeting our inclusion criteria were analyzed. The results indicated that vitamin D supplementation did not significantly reduce pain scores compared to control groups (SMD: -0.130, 95%CI=-0.260 to 0.000; I2=0%), regardless of participants' baseline vitamin D levels. Moreover, long-term supplementation showed no notable improvement in CLBP outcomes (SMD: -0.097, 95%CI=-0.290 to -0.097; I2=19.878%). Additionally, supplementation with active forms of vitamin D (SMD: -0.321, 95%CI=-0.670 to 0.028; I2=0.000%) did not result in significant pain relief for chronic lower back pain. Vitamin D supplementation does not substantially alleviate CLBP. Nevertheless, it may still be considered as part of a comprehensive treatment plan. Further research is necessary to explore its long-term effects and the underlying mechanisms that may explain the observed lack of benefit.

The effect of CD38 inhibition in age-associated vascular dysfunction

Abstract Introduction Aging is characterized by age-related vascular dysfunction, mainly caused by endothelial dysfunction and arterial stiffness. Levels of nicotinamide adenine dinucleotide (NAD) decrease with aging and accordingly, NAD is thought to be involved in the progression of age-related vascular dysfunction. Recent studies reported the involvement of CD38 in NAD regulation; indeed, its inhibition significantly prolongs the lifespan of progeroid and aged mice. However, whether CD38 is involved in the progression of age-related vascular dysfunction remains to be investigated. Methods CD38 expression was investigated in aortic homogenates of young (12-16 weeks) and aged (18-24 months) C57BL/6 wild-type mice, as well as in vitro in young (passage 5-7) and senescent (passage 14-15) primary human aortic endothelial cells (HAECs). Ex vivo tension myograph experiments were performed on thoracic aortas isolated from aged C57BL/6 mice (22-24 months old) pre-incubated for 1 hour with CD38-specific inhibitor, 78c, or DMSO as control. The effect of CD38 inhibition was also observed in vitro in senescent HAECs. Results A significant upregulation of CD38 expression was detected in the aortas of aged mice compared to the young ones as well as in senescent HAECs compared to young cells. Ex vivo tension myograph experiments showed a significant increase in endothelium-dependent relaxation responses to acetylcholine upon CD38 inhibition. The improvement of endothelial function was prevented by the addition of endothelial nitric oxide synthase (eNOS) inhibitor (L-NAME), but not by the COXs inhibitor, indomethacin, indicating that the observed effect achieved by CD38 inhibition is eNOS dependent. The involvement of the eNOS pathway was further supported by the in vitro finding in HAECs of increased eNOS activation upon CD38 inhibition. Conclusion The herein-reported data suggests an involvement of CD38 in the development of age-related endothelial dysfunction. Further studies are ongoing to investigate the underlying molecular mechanisms as well as the effect of long-term CD38 inhibitor supplementation on the progression of age-related vascular dysfunction.

Effect of Long-Term Supplementation of AZOMITE (Hydrated Sodium Calcium Aluminosilicate) in Finishing Diets on Growth Performance, Dietary Energy, and Carcass Yield of Hairy Lambs.

The aim of this study was to evaluate growth performance, dietary energy utilization, and carcass characteristics (carcass weight, dressing percentage, and visceral organ mass) of feedlot lambs fed different levels of AZOMITE (AZO), a source of volcanic clay composed of hydrated sodium calcium aluminosilicate, the same as that included in a finishing diet. For this reason, 36 Pelibuey × Katahdin crossbred intact male lambs (18.81 ± 3.04 kg initial weight) were used in a feeding trial lasting 81 d. Lambs were grouped by initial weight and assigned within six weight groupings to 18 pens in a randomized complete block design. Diets, offered ad libitum, were corn-based finishing diets with an 88:12 concentrate-to-forage ratio supplemented as follows: 1) no AZO inclusion (CTRL), 2) AZO inclusion at a 0.75% level (0.75AZO) in diet dry matter, 3) AZO inclusion at a 1.50% level (1.5AZO) in diet dry matter. Cracked corn was replaced by AZO. After the feeding trial was finished, lambs were slaughtered and carcass weight was registered and dressing percentage was calculated. The data were analyzed as a randomized complete block design, with the pen as the experimental unit. Water consumption and dry matter intake were not affected (p ≥ 0.11) by supplemental AZO. The incorporation of AZO into the diet increased gain efficiency and observed dietary net energy (NE), being maximal with 0.75% AZO inclusion (quadratic effect, p = 0.02). The observed-to-expected dietary NE in the control group was as anticipated (1.00) according to the estimated energy based on the ingredient composition in diet, while in the 0.75AZO group, the observed dietary energy was 6% above expected, indicating a greater efficiency in energy utilization destined to growth (quadratic effect, p = 0.006). Lambs that were fed the AZO treatment showed an improvement (linear effect, p = 0.04) in hot carcass weight (HCW), and tended (linear effect, p = 0.06) to improve dressing percentage (DP) as AZO was increased in the diet. Visceral organ mass was not affected by the treatments. It is concluded that AZOMITE clay can be included up to 1.5% in finishing diets for lambs without negative effects on the intake of water and feed. Lambs that received diets supplemented with 0.75% AZO showed greater improvements in feed efficiency and dietary energy utilization, but an inclusion of 1.50% resulted in greater improvements to HCW and DP. This is the first report regarding the effects of AZO supplementation in the dietary energy utilization of fattening hairy lambs. Further research about the effects of AZO supplementation on ruminal and total tract digestion, carcass and meat quality, and the health of lambs is needed in order to more deeply understand the effects of AZO on its productivity performance.

Long-term food supplementation with sweet basil (Ocimum basilicum L.) prevents age-associated cognitive decline in female mice.

Background: Mild cognitive decline, a common issue in aging, affects memory, learning, and attention. Nutrition can influence cognition, and research indicates that Ocimum sp. (holy basil and sweet basil) leaf extracts may enhance cognition in rodents and humans. However, these studies do not address whether these benefits extend to fresh or dry leaves consumed in typical human diets, along with physiological aging. Aim: To investigate the effects of sweet basil supplementation on cognition in mature and aged female mice. Methods: Female C57bl mice were divided into four groups: 8-month-old mature adults and 18-month-old aged adults, each receiving either a control or supplemented diet. The supplemented diet included a mix of standard chow and fresh basil leaves, administered for 2-8 months. Cognitive and behavioral assessments were conducted using the novel object recognition (NOR), Morris water maze (MWM), and elevated plus maze (EPM) tasks, focusing on memory, learning, and anxiety. Results: No cognitive improvement was observed in mature mice. However, aged mice receiving long-term basil supplementation showed enhanced discrimination in NOR and stayed closer to the absent platform in MWM compared to nonsupplemented controls. While aging mice exhibited reduced anxiety-like behavior in EPM, basil supplementation prevented this reduction. Conclusion: Basil supplementation appears beneficial in elderly mice, potentially preventing age-related cognitive decline and behavioral changes. These findings support the benefits of basil consumption in cognition and underscore its potential role in promoting healthy aging. Incorporating basil into the diet at a younger age may preserve memory and mitigate behavioral changes as individuals age.

Does Colostrum Bovinum Supplementation Affect Swimming Performance in Endurance-Trained Males? A Randomized Placebo-Controlled Crossover Study.

Background/Objectives: Colostrum Bovinum (COL) is recognized for its unique composition and potential ergogenic and immunological benefits. Unlike mature milk, COL is rich in immunoglobulins, lactoferrin, and various growth factors, making it one of the most potent natural immune stimulants. The purpose of this study was to evaluate the effects of 12-weeks of COL supplementation on swimming-specific performance (SSP) and exercise adaptations in endurance-trained male athletes. Methods: Twenty-eight male triathletes and swimmers (age: 31.1 ± 10.2 years; body mass: 81.9 ± 9.0 kg; height: 1.82 ± 0.06 m) participated in a randomized, double-blind, placebo (PLA)-controlled crossover study and received 25 g∙day-1 of COL or PLA for 12 weeks. The study assessed the effects of COL on SSP (8 × 100 m performed at various intensities) and exercise adaptations [heart rate (HR) and blood lactate concentrations ([La-])]. Four main study visits were conducted-before and after COL (COLPRE and COLPOST) and PLA (PLAPRE and PLAPOST) supplementation. Results: COL had no significant effect on SSP. Still, the total time of the SSP test was about ~3.04 s shorter after COL supplementation, and ~7.13 s longer after PLA supplementation. Neither COL nor PLA supplementation affected HR during the SSP test. Post-exercise blood [La-] was significantly reduced after both COL and PLA supplementation. The analysis of SSP results in the consecutive study visits revealed possible existence of the practice effect. Conclusions: Colostrum Bovinum and high-quality milk protein (PLA) seem to be comparably effective in evoking exercise adaptation in endurance-trained male athletes. Long-term crossover supplementation protocols in athletes must consider the impact of possible practice effect when interpreting the outcomes related to exercise performance, but not biochemical or physiological markers of exercise adaptation.

PSXIII-27 Effects of long-term dietary supplementation of nitrate on enteric methane production and performance of Hanwoo steers

Abstract Nitrate (NO3-) has been recognized as an effective dietary additive to reduce enteric methane (CH4) production in ruminants. Two experiments were performed to evaluate the effects of long-term supplementation and method of supplementing calcium ammonium nitrate (CAN) on enteric CH4 production in Hanwoo steers. In Exp. 1, Hanwoo steers [n = 20; body weight (BW) = 348 ± 18 kg] were divided into two groups of 10, and were assigned to one of the two diets 1) CON, 500 g/kg timothy hay and 500 g/kg commercial concentrate, or 2) CAN, basal diet top-dressed with 2.5% CAN [1.578% NO3- in the dietary dry matter (DM)] for 115 d in a completely randomized design. Measurement of CH4 was performed for 3 d starting from d 21, d 51, d 80 and d 112 using whole body respiratory chambers (RC). Rumen fluid and blood samples were collected on d 25 and d 26 of the feeding trial, respectively. In Exp. 2, the animals were re-grouped (476 ± 21 kg) and fed the same basal diet supplemented with 2.0% CAN (1.262% NO3- in the dietary DM) incorporated in the concentrate pellet (P-CAN). Methane was measured on d 19 and d 45 for 3 d. In Exp. 1, CAN supplementation decreased (P &amp;lt; 0.05) DM intake (DMI) of the steers both in the barn and RC. However, no negative effects on average daily gain or feed conversion ratio were observed. Also, no potential toxicity of NO3- was observed based on blood methemoglobin concentrations (all samples &amp;lt; 2.5% of total hemoglobin). A decrease (P &amp;lt; 0.05) in CH4 production (g/d) caused by decreased DMI was noted in CAN throughout the feeding trial. However, CH4 yield (g/kg DMI) decreased (P &amp;lt; 0.05) only in the beginning phase (d 21) and did not differ (P &amp;gt; 0.05) thereafter. Ruminal pH, ammonia and total volatile fatty acid concentration did not differ between treatments (P &amp;gt; 0.05), but CAN increased and decreased (P &amp;lt; 0.0001) acetate and butyrate proportions, respectively. Steers fed CAN had greater ruminal abundance of Ruminococcus sp. (P &amp;lt; 0.05), Succiniclasticum (P = 0.066), and Selenomonas ruminantium (P = 0.073), but a reduced (P &amp;lt; 0.05) abundance of Methanobrevibacter ruminantium (P = 0.073). In Exp. 2, P-CAN did not exert any effects on DMI, CH4 production, CH4 yield and other production parameters. Overall, CAN can be used as a feed additive for beef cattle in the long-term without NO3- intoxication or any negative effects on growth performance. However, lack of persistent decrease in CH4 yield suggested the need of further evaluation of CAN under different dietary conditions. Furthermore, incorporating CAN in the concentrate pellet can be a suitable approach, since top-dressing may alter the organoleptic properties of diets that may reduce DMI of the steers.

High-output cardiac failure in the setting of iron deficiency anaemia: a case report.

We present a case of a child with high-output heart failure and severe iron deficiency anaemia-induced dilated cardiomyopathy managed with serial blood transfusions, preload and afterload reducing agents, inotropic therapies, and long-term iron supplementation. The complete resolution of echocardiogram findings of moderate enlargement of all cardiac chambers and moderately depressed left ventricular systolic function was achieved.

Acute and prolonged effects of Bacillus amyloliquefaciens GF424-derived SOD on antioxidant defense in healthy individuals challenged with intense aerobic exercise

Reactive oxygen species (ROS) play a vital role in cellular functions but can lead to oxidative stress and contribute to degenerative diseases when produced in excess. Maintaining redox balance is essential and can be achieved through innate defense mechanisms or external antioxidants. Superoxide dismutase (SOD) is a key enzyme that mitigates intracellular oxidative stress by converting harmful free radicals into hydrogen peroxide, which is subsequently neutralized by catalase and glutathione peroxidase. Previous studies have demonstrated the antioxidant capabilities of SOD derived from Bacillus amyloquefaciens GF424 (BA-SOD) in murine models exposed to either irradiation or SOD1 gene deletion. In this study, a randomized clinical trial was conducted to evaluate the antioxidative benefits of BA-SOD in healthy individuals undergoing acute aerobic exercise (AAE). Eighty participants were randomly assigned to receive either BA-SOD or a placebo for 8 weeks. Antioxidant enzyme activities and glutathione levels were measured before, immediately after, and 30 min post-exercise. A single dose of BA-SOD significantly reduced ROS levels induced by AAE, primarily by enhancing SOD activity in erythrocytes and activating glutathione peroxidase. Continuous BA-SOD administration was associated with a sustained increase in catalase activity and elevated levels of reduced glutathione (GSH). Transcriptomic and metabolomic analyses revealed that a single BA-SOD dose facilitated GSH oxidation, as evidenced by decreased levels of serine, glutamine, and glycine, and increased pyroglutamate levels. Additionally, repeated dosing led to increased expression of genes encoding isocitrate dehydrogenase and malic enzyme, which are involved in NADPH synthesis, as well as nicotinamide phosphoribosyl transferase and NAD kinase, which are essential for NADP availability–critical for converting oxidized glutathione (GSSG) back to GSH. These molecular insights align with clinical observations, suggesting that both acute and long-term BA-SOD supplementation may effectively enhance antioxidant defenses and maintain redox balance under oxidative stress conditions.

Effects of long-term magnesium L-threonate supplementation on neuroinflammation, demyelination and blood–brain barrier integrity in mice with neuromyelitis optica spectrum disorder

In clinical practice, we found cerebrospinal fluid magnesium concentration significantly lower in neuromyelitis optica spectrum disorder (NMOSD) patients compared to controls with non-autoimmune encephalitis neurological diseases. To investigate the effects and potential mechanisms of long-term magnesium supplementation on neuroinflammation, demyelination, and blood–brain barrier (BBB) integrity in NMOSD, we used two models: (1) NMOSD mouse model, which was induced by intraperitoneal injection of purified NMO-IgG to experimental autoimmune encephalomyelitis (EAE) mice, and (2) cultured human cerebral microvascular endothelial cells/D3 (hCMEC/D3). In the NMOSD mouse model, Magnesium L-threonate (MgT) pretreatment alleviated NMO-IgG–induced effects, including AQP4 loss, leukocyte infiltration, astrocyte and microglia activation, demyelination, decreased tight junction (TJ) protein expression, and neurological deficits. In vitro, MgT pretreatment ameliorated NMO-IgG induced damage to TJ protein expression in a (transient receptor potential melastatin 7) TRPM7-dependent manner. Magnesium supplementation shows potential protective effects against NMOSD, suggesting it may be a novel therapeutic approach for this condition. The beneficial effects appear to be mediated through preservation of blood–brain barrier integrity and reduction of neuroinflammation and demyelination.

Long-term d-allose administration ameliorates age-related cognitive impairment and loss of bone strength in male mice

Age-related physical and cognitive decline may be ameliorated by consuming functional foods. d-Allose, reported to have multiple health benefits, may temper aging phenotypes, particularly brain function. We investigated whether d-allose supplementation improves cognitive function. A standard battery of behavioral tests was administered to 18-month-old male mice after consuming diet containing 3 % d-allose for 6 months. Following a wire-hanging test, an open-field test, Morris water maze, fear-conditioning, and an analgesia test were sequentially performed. Bone density and strength were assessed afterwards. Possible mechanism(s) under-lying memory changes in hippocampus were also examined with a DNA microarray. d-Allose failed to influence muscle strength, locomotor activity and anxiety, fear memory, or pain sensitivity. However, d-allose improved hippocampus-dependent spatial learning and memory, and it may contribute to increase bone strength. d-Allose also changed the expression of some genes in hippocampus involved in cognitive functions. Long-term d-allose supplementation appears to modestly change aging phenotypes and improve spatial memory.

Long-term GABA Supplementation Regulates Diabetic Gastroenteropathy through GABA Receptor/trypsin-1/PARs/Akt/COX-2 Axis.

Molecular alterations of diabetic gastroenteropathy are poorly identified. This study investigates the effects of prolonged GABA supplementation on key protein expression levels of trypsin-1, PAR-1, PAR-2, PAR-3, PI3K, Akt, COX-2, GABAA, and GABAB receptors in the gastric tissue of type 2 diabetic rats (T2DM). To induce T2DM, a 3-month high-fat diet and 35 mg/kg of streptozotocin was used. Twenty-four male Wistar rats were divided into 4 groups: (1) control, (2) T2DM, (3) insulin-treated (2.5 U/kg), and (4)GABA-treated (1.5 g/kg GABA). Blood glucose was measured weekly. The protein expressions were assessed using western blotting. Histopathological changes were examined by H&E and Masson's staining. Diabetic rats show reduced NOS1 and elevated COX-2 and trypsin-1 protein expression levels in gastric tissue. Insulin and GABA therapy restored the NOS1 and COX-2 levels to control values. Insulin treatment increased PI3K, Akt, and p-Akt and, decreased trypsin-1, PAR-1, PAR-2, and PAR-3 levels in the diabetic rats. Levels of GABAA and GABAB receptors normalized following insulin and GABA therapy. H&E staining indicated an increase in mucin secretion following GABA treatment. These results suggest that GABA by acting on GABA receptors may regulate the trypsin-1/PARs/Akt/COX-2 pathway and thereby improve complications of diabetic gastroenteropathy.

Effect of Selol on Tumor Morphology and Biochemical Parameters Associated with Oxidative Stress in a Prostate Tumor-Bearing Mice Model.

Prostate cancer is the leading cause of cancer death in men. Some studies suggest that selenium Se (+4) may help prevent prostate cancer. Certain forms of Se (+4), such as Selol, have shown anticancer activity with demonstrated pro-oxidative effects, which can lead to cellular damage and cell death, making them potential candidates for cancer therapy. Our recent study in healthy mice found that Selol changes the oxidative-antioxidative status in blood and tissue. However, there are no data on the effect of Selol in mice with tumors, considering that the tumor itself influences this balance. This research investigated the impact of Selol on tumor morphology and oxidative-antioxidative status in blood and tumors, which may be crucial for the formulation's effectiveness. Our study was conducted on healthy and tumor-bearing animal models, which were either administered Selol or not. We determined antioxidant enzyme activities (Se-GPx, GPx, GST, and TrxR) spectrophotometrically in blood and the tumor. Furthermore, we measured plasma prostate-specific antigen (PSA) levels, plasma and tumor malondialdehyde (MDA) concentration as a biomarker of oxidative stress, selenium (Se) concentrations and the tumor ORAC value. Additionally, we assessed the impact of Selol on tumor morphology and the expression of p53, BCL2, and Ki-67. The results indicate that treatment with Selol influences the morphology of tumor cells, indicating a potential role in inducing cell death through necrosis. Long-term supplementation with Selol increased antioxidant enzyme activity in healthy animals and triggered oxidative stress in cancer cells, activating their antioxidant defense mechanisms. This research pathway shows promise in understanding the anticancer effects of Selol. Selol appears to increase the breakdown of cancer cells more effectively in small tumors than in larger ones. In advanced tumors, it may accelerate tumor growth if used as monotherapy. Therefore, further studies are necessary to evaluate its efficacy either in combination therapy or for the prevention of recurrence.

Dietary nitrate maintains intestinal epithelia homeostasis in aged mice.

The intestinal tract, which is the primary site of digestion and absorption of nutrients, is one of the most vulnerable organs during aging. Dietary nitrate, which is mainly derived from the diet and absorbed in the intestinal tract, is a key messenger that connecting oral and general health. However, whether dietary nitrate regulates intestinal tract homeostasis remains unclear. Our data revealed that the serum and salivary nitrate levels decreased during mice aging. The functional proteins of the epithelial barrier (E-cadherin, Claudin-1 and Zonula Occludens-1) in the colon tissues decreased during the aging process. Long-term nitrate supplement in drinking water restored the serum and salivary nitrate levels and increased the functional proteins expression of the colon epithelial barrier. Dietary nitrates increase the relative abundance of some intestinal probiotics, particularly those associated with the production of short-chain fatty acids, such as Blautia, Alloprevotella, Butyricicoccus, and Ruminococcaceae, while promoting the butyric acid production in the colon. Moreover, the expression of Sialin (encoded by Slc17a5), which is a nitrate transporter, increased in the colon epithelial cells by nitrate supplementation. The epithelial cell-conditional Slc17a5-knockout mutant mice (K14-cre; Slc17a5fl/fl) revealed that the functional proteins expression of the colon epithelial barrier and the proliferation of PCNA-positive intestinal epithelial cells in the colon crypts was significantly decreased compared with those of the K14-cre; Slc17a5fl/+ mice. Taken together, our findings suggested that nitrate supplementations were associated with the increased expression of colonic epithelial barriers-related proteins and the increased Sialin expression. Nitrate may serve as a potential therapeutic approach in maintaining aged colonic homeostasis.

Long-chain n-3 polyunsaturated fatty acid supplementation and neuromuscular function in older adults.

This review aims to explore the latest research investigating the effects of marine-derived long-chain n -3 polyunsaturated fatty acid (LC n -3 PUFA) supplementation on neuromuscular function in older adults. Ageing results in a decline in skeletal muscle strength and mass. There is growing evidence that LC n -3 PUFA supplementation increases muscle strength and mass in healthy older adults, yet the mechanisms underlying these effects remain elusive. Recent studies investigating LC n -3 PUFA supplementation have demonstrated effects on neuromuscular function such as increases in the compound muscle action potential (M-wave) amplitude and surface electromyography alongside increases in muscular strength. Therefore, evidence suggests that LC n -3 PUFA may elicit a beneficial effect at the neuromuscular junction and possess neuroprotective properties in older adults. LC n -3 PUFA supplementation may increase or maintain neuromuscular function throughout the ageing process. Further research is warranted to investigate the long-term effects LC n -3 PUFA supplementation on neuromuscular outcomes such as single motor unit properties and cortical/supraspinal networks, utilizing state-of-the-art techniques in neuromuscular physiology.

Abstract We038: Metal-organic frameworks as next-generation cardiac therapeutics

Background: Zinc (Zn) is an essential trace element for the normal functioning of the heart. Long-term dietary supplementation of Zn has been shown to improve cardiac function and prevent myocardial damage caused by ischemia-reperfusion injury. However, the major limitation in delivering Zn to the cardiac tissue is the off-targeted absorption and the duration of treatment to achieve a therapeutic effect. Hypothesis: Cardiac-targeted delivery of trace elements like Zn may attenuate myocardial damage caused by ischemia-reperfusion injury, improve cardiac function, and exert a cardioprotective effect. Methods: Zn-based metal-organic frameworks (ZIF-NPs) were fabricated and tested on Microvascular Endothelial Cells (MVECs), and human-induced pluripotent derived cardiomyocytes (hiCMs) for cytoprotective effect against hypoxic stress using Live/dead assay and multi-electrode array (MEA) analysis respectively. The in vivo efficacy of ZIF-NPs was tested in the rodent myocardial infarction (MI) model and cardiac function was assessed via M-mode echocardiography. Results: The fabricated ZIF-NPs were biocompatible and showed significant intracellular uptake causing the induction of cytoprotective metallothioneins. ZIF-NP treatment showed enhanced cell survival in serum-starved MVECs and exhibited pro-angiogenic effects in vitro. Similarly, hiCMs treated with ZIF-NPs displayed enhanced endurance and electrophysiological function, when subjected to hypoxic stress and protected the cells against ROS-induced DNA damage. Intramyocardial delivery of ZIF-NPs in the cardiac tissue improved cardiac function and attenuated fibrosis post-MI. Conclusions: ZIF-NPs exerted a pro-survival, cytoprotective response in both in vitro and in vivo conditions. Our study highlights the significance of cardiac targeted Zn delivery by ZIF-NPs for cardioprotection as next-generation cardiac therapeutics.