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Nutrients Jun 2024Beef is an important source of high-quality protein and several micronutrients, including iron, zinc, and B vitamins. We determined beef intake and its relationship with...
Beef Consumption Is Associated with Higher Intakes and Adequacy of Key Nutrients in Older Adults Age 60+ Years: National Health and Nutrition Examination Survey 2011-2018 Analysis.
Beef is an important source of high-quality protein and several micronutrients, including iron, zinc, and B vitamins. We determined beef intake and its relationship with intakes of nutrients and their adequacy using 24 h dietary recall data from 5868 older adults. Usual intakes from foods were determined using the National Cancer Institute method, and the percent of the population below the estimated average requirement or above adequate intake was estimated. A high percentage of older adults did not meet nutrient recommendations for vitamin D (96%), choline (96%), vitamin E (84%), potassium (70%), calcium (63%), magnesium (60%), vitamin C (46%), vitamin A (39%), zinc (21%), vitamin B (19%), and folate (15%). About 68% of older adults were beef consumers with a mean intake of 56 g/day. Beef consumers had higher ( < 0.05) intakes of energy, protein, calcium, iron, phosphorus, selenium, sodium, zinc, thiamin, riboflavin, niacin, vitamin B, and choline, and a higher ( < 0.05) proportion met nutrient recommendations for protein, calcium, copper, zinc, thiamin, folate, and vitamin B than non-consumers. Consumers of fresh, ground, and processed beef also had generally higher intakes and lower inadequacies of many nutrients depending on the beef type. In conclusion, older adults generally had poor nutrient adequacy from their diets, while beef consumers had higher nutrient intakes and adequacy for certain key nutrients, which are inherently generally available from beef or from foods consumed with beef.
Topics: Humans; Aged; Male; Female; Middle Aged; Nutrition Surveys; Red Meat; Diet; Nutrients; Nutritional Status; Micronutrients; United States; Aged, 80 and over; Recommended Dietary Allowances; Nutritive Value; Cattle; Animals; Nutritional Requirements
PubMed: 38892711
DOI: 10.3390/nu16111779 -
Nature Communications Jun 2024Methylenetetrahydrofolate reductase (MTHFR) is a pivotal flavoprotein connecting the folate and methionine methyl cycles, catalyzing the conversion of...
Methylenetetrahydrofolate reductase (MTHFR) is a pivotal flavoprotein connecting the folate and methionine methyl cycles, catalyzing the conversion of methylenetetrahydrofolate to methyltetrahydrofolate. Human MTHFR (hMTHFR) undergoes elaborate allosteric regulation involving protein phosphorylation and S-adenosylmethionine (AdoMet)-dependent inhibition, though other factors such as subunit orientation and FAD status remain understudied due to the lack of a functional structural model. Here, we report crystal structures of Chaetomium thermophilum MTHFR (cMTHFR) in both active (R) and inhibited (T) states. We reveal FAD occlusion by Tyr361 in the T-state, which prevents substrate interaction. Remarkably, the inhibited form of cMTHFR accommodates two AdoMet molecules per subunit. In addition, we conducted a detailed investigation of the phosphorylation sites in hMTHFR, three of which were previously unidentified. Based on the structural framework provided by our cMTHFR model, we propose a possible mechanism to explain the allosteric structural transition of MTHFR, including the impact of phosphorylation on AdoMet-dependent inhibition.
Topics: Methylenetetrahydrofolate Reductase (NADPH2); S-Adenosylmethionine; Allosteric Regulation; Chaetomium; Phosphorylation; Humans; Crystallography, X-Ray; Models, Molecular; Flavin-Adenine Dinucleotide
PubMed: 38886362
DOI: 10.1038/s41467-024-49327-5 -
ACS Omega Jun 2024Schistosomiasis is a neglected disease of poverty that affects over 200 million people worldwide and relies on a single drug for therapy. The cathepsin B1 cysteine...
Schistosomiasis is a neglected disease of poverty that affects over 200 million people worldwide and relies on a single drug for therapy. The cathepsin B1 cysteine protease (SmCB1) of has been investigated as a potential target. Here, a structure-based pharmacophore virtual screening (VS) approach was used on a data set of approved drugs to identify potential antischistosomal agents targeting SmCB1. Pharmacophore (PHP) models underwent validation through receiver operating characteristics curves achieving values >0.8. The data highlighted riboflavin (RBF) as a compound of particular interest. A 1 μs molecular dynamics simulation demonstrated that RBF altered the conformation of SmCB1, causing the protease's binding site to close around RBF while maintaining the protease's overall integrity. RBF inhibited the activity of SmCB1 at low micromolar values and killed the parasite in vitro. Finally, in a murine model of infection, oral administration of 100 mg/kg RBF for 7 days significantly decreased worm burdens by ∼20% and had a major impact on intestinal and fecal egg burdens, which were decreased by ∼80%.
PubMed: 38882094
DOI: 10.1021/acsomega.4c03376 -
Journal of Biomedical Optics Jun 2024To enable non-destructive longitudinal assessment of drug agents in intact tumor tissue without the use of disruptive probes, we have designed a label-free method to...
SIGNIFICANCE
To enable non-destructive longitudinal assessment of drug agents in intact tumor tissue without the use of disruptive probes, we have designed a label-free method to quantify the health of individual tumor cells in excised tumor tissue using multiphoton fluorescence lifetime imaging microscopy (MP-FLIM).
AIM
Using murine tumor fragments which preserve the native tumor microenvironment, we seek to demonstrate signals generated by the intrinsically fluorescent metabolic co-factors nicotinamide adenine dinucleotide phosphate [NAD(P)H] and flavin adenine dinucleotide (FAD) correlate with irreversible cascades leading to cell death.
APPROACH
We use MP-FLIM of NAD(P)H and FAD on tissues and confirm viability using standard apoptosis and live/dead (Caspase 3/7 and propidium iodide, respectively) assays.
RESULTS
Through a statistical approach, reproducible shifts in FLIM data, determined through phasor analysis, are shown to correlate with loss of cell viability. With this, we demonstrate that cell death achieved through either apoptosis/necrosis or necroptosis can be discriminated. In addition, specific responses to common chemotherapeutic treatment inducing cell death were detected.
CONCLUSIONS
These data demonstrate that MP-FLIM can detect and quantify cell viability without the use of potentially toxic dyes, thus enabling longitudinal multi-day studies assessing the effects of therapeutic agents on tumor fragments.
Topics: Animals; Mice; Cell Survival; Microscopy, Fluorescence, Multiphoton; Apoptosis; Flavin-Adenine Dinucleotide; NADP; Cell Line, Tumor; Optical Imaging
PubMed: 38881557
DOI: 10.1117/1.JBO.29.S2.S22709 -
Frontiers in Nutrition 2024This study investigated the efficacy of a mixed beet-based supplement (BEET) versus placebo (PL) in countering inflammation during recovery from 2.25 h of intensive...
OBJECTIVES
This study investigated the efficacy of a mixed beet-based supplement (BEET) versus placebo (PL) in countering inflammation during recovery from 2.25 h of intensive cycling in 20 male and female cyclists. A multi-omics approach was used that included untargeted proteomics and a targeted oxylipin panel.
METHODS
A randomized, placebo-controlled, double-blind, crossover design was used with two 2-week supplementation periods and a 2-week washout period. Supplementation periods were followed by a 2.25 h cycling bout at close to 70%VO. The BEET supplement provided 212 mg of nitrates per day, 200 mg caffeine from green tea extract, 44 mg vitamin C from Camu Camu berry, B-vitamins from quinoa sprouts (40% Daily Value for thiamin, riboflavin, niacin, and vitamin B6), and 2.5 g of a mushroom blend containing Cordyceps sinensis and Inonotus obliquus. Six blood samples were collected before and after supplementation (overnight fasted state), immediately post-exercise, and at 1.5 h-, 3 h-, and 24 h-post-exercise.
RESULTS
The 2.25 h cycling bout increased plasma levels of 41 of 67 oxylipins detected. BEET supplementation significantly increased plasma nitrate (NO ) and nitrite (NO ) (sum, NO + NO ) concentrations (interaction effect, < 0.001) and two anti-inflammatory oxylipins [18-hydroxyeicosapentaenoic acid (18-HEPE) and 4-hydroxy-docosahexanoic acid (4-HDoHE)]. The untargeted proteomics analysis identified 616 proteins (458 across all times points), and 2-way ANOVA revealed a cluster of 45 proteins that were decreased and a cluster of 21 that were increased in the BEET versus PL trials. Functional enrichment supported significant BEET-related reductions in inflammation-related proteins including several proteins related to complement activation, the acute phase response, and immune cell adhesion, migration, and differentiation.
DISCUSSION
Intake of a BEET-based supplement during a 2-week period was linked to higher plasma levels of NO + NO , elevated post-exercise levels of two anti-inflammatory oxylipins, and a significant decrease in a cluster of proteins involved in complement activation and inflammation. These data support that 2-weeks intake of nitrate from a mixed beet-based supplement moderated protein biomarkers of exercise-induced inflammation in athletes.
PubMed: 38873567
DOI: 10.3389/fnut.2024.1408804 -
Frontiers in Veterinary Science 2024Garlic skin (GAS) has been proven to improve the growth performance of fattening sheep. However, the mechanism by which GAS affects fattening sheep is not yet clear. The...
OBJECTIVE
Garlic skin (GAS) has been proven to improve the growth performance of fattening sheep. However, the mechanism by which GAS affects fattening sheep is not yet clear. The aim of this study is to investigate the effects of adding GAS to feed on the growth performance, rumen and fecal microbiota, serum and urine metabolism, and transcriptomics of rumen epithelial cells in fattening sheep.
METHODS
GAS with 80 g/kg dry matter (DM) was added to the diet of fattening sheep to study the effects of GAS on gut microbiota, serum and urine metabolism, and transcriptome of rumen epithelial tissue in fattening sheep. Twelve Hu sheep (body weights; BW, 23.0 ± 2.3 kg and ages 120 ± 3.5 d) were randomly divided into two groups. The CON group was the basal diet, while the GAS group was supplemented with GAS in the basal diet. The trial period was 10 weeks, with the first 2 weeks being the pre-trial period.
RESULTS
The daily average weight gain of fattening sheep in the GAS group was significantly higher than that in the CON group ( < 0.05), and the serum GSH-Px of the GAS group fattening sheep was significantly increased, while MDA was significantly reduced ( < 0.05). Based on the genus classification level, the addition of garlic peel in the diet changed the intestinal microbial composition, and the relative abundance was significantly upregulated by ( < 0.05), while significantly downregulated by , , and ( < 0.05). Metabolomics analysis found that there were 166 significantly different metabolites in serum and 68 significantly different metabolites in urine between the GAS and CON groups ( < 0.05). GAS had an impact on amino acid metabolism, pyrimidine metabolism, methane metabolism, riboflavin metabolism, and unsaturated fatty acid synthesis pathways ( < 0.05). Transcriptome sequencing showed that differentially expressed genes were mainly enriched in immune regulatory function, improving the health of fattening sheep.
CONCLUSION
Adding GAS can improve the energy metabolism and immune function of fattening sheep by altering gut microbiota, metabolome, and transcriptome, thereby improving the growth performance of fattening sheep.
PubMed: 38872796
DOI: 10.3389/fvets.2024.1409518 -
Frontiers in Bioengineering and... 2024Previous laboratory-scale studies have consistently shown that carbon-based conductive materials can notably improve the anaerobic digestion of food waste, typically...
Previous laboratory-scale studies have consistently shown that carbon-based conductive materials can notably improve the anaerobic digestion of food waste, typically employing reactors with regular capacity of 1-20 L. Furthermore, incorporating riboflavin-loaded conductive materials can further address the imbalance between fermentation and methanogenesis in anaerobic systems. However, there have been few reports on pilot-scale investigation. In this study, a 10 m of riboflavin modified carbon cloth was incorporated into a pilot-scale (2 m) food waste anaerobic reactor to improve its treatment efficiency. The study found that the addition of riboflavin-loaded carbon cloth can increase the maximum organic loading rate (OLR) by 40% of the pilot-scale reactor, compared to the system using carbon cloth without riboflavin loading, while ensuring efficient operation of the reaction system, effectively alleviating system acidification, sustaining methanogen activity, and increasing daily methane production by 25%. Analysis of the microbial community structure revealed that riboflavin-loaded carbon cloth enriched the methanogenic archaea in the genera of and , which are capable of extracellular direct interspecies electron transfer (DIET). And metabolic pathway analysis identified the methane production pathway, highly enriched on the reduction of acetic acid and CO at riboflavin-loaded carbon cloth sample. The expression levels of genes related to methane production via DIET pathway were also significantly upregulated. These results can provide important guidance for the practical application of food waste anaerobic digestion engineering.
PubMed: 38863495
DOI: 10.3389/fbioe.2024.1395810 -
Turkish Journal of Ophthalmology Jun 2024To investigate the clinical efficacy and safety of the modified Cretan protocol in patients with post-laser in situ keratomileusis ectasia (PLE).
OBJECTIVES
To investigate the clinical efficacy and safety of the modified Cretan protocol in patients with post-laser in situ keratomileusis ectasia (PLE).
MATERIALS AND METHODS
In this retrospective study, 26 eyes of 16 patients with PLE were treated with the modified Cretan protocol (combined transepithelial phototherapeutic keratectomy and accelerated corneal collagen cross-linking). Visual, refractive, tomographic, and aberrometric outcomes and point spread function (PSF) were recorded preoperatively and at 6, 12, and 24 months after treatment.
RESULTS
Both uncorrected and best corrected visual acuity were stable at 24 months postoperatively compared to baseline (from 0.89±0.36 to 0.79±0.33 logarithm of the minimum angle of resolution [LogMAR] and 0.31±0.25 to 0.24±0.19 LogMAR, respectively, p>0.05 for all values). The mean K1, K2, Kmean, thinnest corneal thickness, and spherical aberration at baseline were 45.76±5.75 diopters (D), 48.62±6.17 D, 47.13±5.89 D, 433.16±56.86 μm, and -0.21±0.63 μm respectively. These values were reduced to 42.86±6.34 D, 45.92±6.74 D, 44.21±6.4 D, 391.07±54.76 μm, and -0.51±0.58 μm at 24 months postoperatively (p<0.001, p=0.002, p<0.001, p=0.001, and p=0.02, respectively). The mean spherical equivalent, manifest cylinder, Kmax, central corneal thickness, other corneal aberrations (root mean square, trefoil, coma, quatrefoil, astigmatism), and PSF remained stable (p>0.05 for all variables), while anterior and posterior elevation were significantly improved at 24 months postoperatively (p<0.001 and p=0.02, respectively). No surgical complications occurred during the 24-month follow-up.
CONCLUSION
The modified Cretan protocol is a safe and effective treatment option for PLE patients that provides visual stabilization and significant improvement in topographic parameters during the 24-month follow-up. Further studies are needed to support our results.
Topics: Humans; Retrospective Studies; Keratomileusis, Laser In Situ; Male; Female; Visual Acuity; Adult; Dilatation, Pathologic; Corneal Topography; Refraction, Ocular; Cross-Linking Reagents; Treatment Outcome; Photosensitizing Agents; Young Adult; Collagen; Lasers, Excimer; Follow-Up Studies; Riboflavin; Photochemotherapy; Corneal Diseases; Cornea; Postoperative Complications; Myopia; Ultraviolet Rays
PubMed: 38853628
DOI: 10.4274/tjo.galenos.2024.82342 -
Se Pu = Chinese Journal of... Jun 2024Mitochondria perform various metabolic processes that significantly affect cell differentiation, proliferation, signal transduction, and programmed cell death. The...
Mitochondria perform various metabolic processes that significantly affect cell differentiation, proliferation, signal transduction, and programmed cell death. The disruption of mitochondrial bioenergetic and metabolic functions is closely related to many disorders. The specific isolation and purification of intact, high-purity, and functional mitochondria are central to the understanding of their mechanism of action but remain challenging tasks. In this study, a mitochondrial penetrating peptide (MPP) with the sequence FrFKFrFK(Ac) was used as a mitochondrial recognition motif to construct a peptide-guided affinity separation material. The multiple aromatic phenylalanine (F) residues in this amphiphilic peptide can confer lipophilicity to the mitochondrial membrane, whereas the basic residues (D-arginine and lysine) render the MPP surface positively charged, thereby promoting the binding of negatively charged mitochondria. After the derivatization of the N terminal of MPP with an oligoglycine spacer, the peptide ligands were conjugated to matrix beads (MB) with surface aldehyde functional groups. Peptide functionalization was performed via a condensation reaction between the amino group in the peptide ligand and the aldehyde group on the beads. The generated Schiff bases were reduced, affording stable covalent bonds. The dense and stable functionalization of the beads with the mitochondria-targeting peptides was demonstrated using high performance liquid chromatography (HPLC), zeta potential assay, and scanning electron microscopy (SEM). The immobilization efficiency of the peptide ligands was 1.47 μmol/g, and the surface potential of MB@MPP was 11 mV. MB@MPP was used for the direct isolation of mitochondria after cell homogenization. As observed by SEM, mitochondria with a cross-sectional diameter of 500 nm were efficiently captured on the MB@MPP surface. Because the mitochondrial membrane potential is an important marker of mitochondrial function and the driving force behind the staining of mitochondria with Mito Tracker dyes, the specific binding and separation of fluorescent mitochondria from the cell samples revealed that the proposed MB@MPP-based isolation approach can keep mitochondria intact and retain their functions. Western blot assays were employed to characterize the protein markers of the mitochondria (citrate synthase (CS) and voltage-dependent anion channel protein (VDAC)) and cytoplasmic protein (vinculin), and examine the integrity and purity of the captured mitochondria. The results showed that the lysates released from MB@MPP had high CS and VDAC contents. By contrast, vinculin, which is highly abundant in whole-cell lysates, was barely detected in the lysates from MB@MPP. These results suggest that MB@MPP isolates mitochondria with high affinity, specificity, and antifouling ability by using the targeting peptide as the capture handle. A comparison with a commercial mitochondrial isolation kit demonstrated that MB@MPP can separate mitochondria with higher CS and VDAC abundance and purity. Given the superior separation performance of MB@MPP, the molecular profiles of the isolated mitochondria under stress were subjected to further analysis of their molecular profiles under stress. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established to detect tryptophan (Trp) and riboflavin in the mitochondria. Quantification was performed in multiple-reaction monitoring (MRM) mode. Owing to the high purity of the mitochondria, the Trp and riboflavin contents were determined to be 265 and 0.67 nmol/mg, respectively. The metabolic response of mitochondria to external stimuli was further examined using acadesine, an adenosine 5'-monophosphate (AMP)-activated protein kinase activator with a wide range of metabolic effects, to treat cells. After cell homogenization, MB@MPP was used to separate the mitochondria from the cell samples with and without acadesine treatment, followed by LC-MS/MS analysis. The quantification results demonstrated that acadesine induced a 14% upregulation of Trp content in the mitochondria. By contrast, the riboflavin content decreased to 0.48 nmol/mg, which is 72% of that in untreated mitochondria. The changes in Trp and riboflavin contents could influence their metabolic pathways and, thus, the levels of their metabolites, such as nicotinamide adenine dinucleotide, flavin mononucleotide, and flavin adenine dinucleotide, which are essential coenzymes in mitochondria. Peptide-functionalized affinity microbeads with high affinity and specificity for mitochondria are promising for the efficient isolation of high-quality mitochondria, and offer a useful tool for understanding the complicated functions and dynamics of this unique organelle.
Topics: Mitochondria; Peptides; Animals; Chromatography, Affinity
PubMed: 38845516
DOI: 10.3724/SP.J.1123.2024.01013