Pantothenic Research Articles

Citrus is a Multivitamin Treasure Trove: A Review

Citrus is popularly known as the source of beneficial and essential nutrients for human health, including vitamins. The current review revealed the content of multivitamins, not only vitamin C but also vitamins A, B, and E that are not widely acknowledged within Citrus. Numerous Citrus genotypes contain vitamin C, with the grapefruit (Citrus paradisi) being the richest, and citron (C. medica) the poorest. Vitamin A in the form of β-carotene, α-carotene, and β-cryptoxanthin is commonly found within Citrus, especially in several colored flesh species such as grapefruit, mandarin (C. reticulate), and orange (C. sinensis). In terms of vitamin B, orange and grapefruit are proven to contain B-complex, including thiamine (B1), riboflavin (B2), niacin (B3), pantothenic acid (B5), pyridoxine (B6), biotin (B7), inositol (B8) and folate (B9). Vitamin E in the form of α-tocopherol was detected in leaf kaffir lime (C. hystrix) and orange (C. sinensis), lemon (C. limon), mandarin (C. reticulate), and tangerine (C. nobilis) fruit. This review summarizes the nutritional content of Citrus; Citrus contains not only vitamin C but also other vitamins beneficial to human health, therefore Citrus consumption is highly recommended.

Metabolomic, oxidative, and inflammatory responses to acute exercise in chronic obstructive pulmonary disease

BackgroundThere is currently a need to identify metabolomic responses to acute exercise in chronic obstructive pulmonary disease (COPD). ObjectiveWe investigated the metabolomic, oxidative, and inflammatory responses to constant (CE) and intermittent (IE) work rate exercises in COPD. MethodsSixteen males with COPD performed a symptom-limited incremental cycle exercise test (ICE). Metabolomic, oxidative, and inflammatory responses to CE and IE (based on the performance of ICE) were analyzed in the plasma. ResultsFructose-6-phosphate, 3-phosphoglyceric acid, l-carnitine, and acylcarnitines levels were significantly decreased, whereas alpha-ketoglutaric, malic, 2-hydroxybutyric, and 3-hydroxybutyric acids were increased, after CE and IE (p<0.05). Increases in citric, isocitric, and lactic acids, as well as decreases in pyruvic and oxalic acids, were only present with IE (p<0.05). Isoleucine was decreased after both exercises (p<0.05). We observed an increase in inosine-5′-diphosphate, uric acid, ascorbic acid, and pantothenic acid, as well as a decrease in 5-hydroxymethyluridine, threonic acid, and dehydroascorbic acid, after IE (p<0.05). Catalase, reduced glutathione, and total antioxidant status difference values for both exercises were similar (p>0.05). The change in glutathione peroxidase (GPx) with CE was more significant than that with IE (p = 0.004). The superoxide dismutase change was greater with IE than with CE (p = 0.015). There were no significant changes in inflammatory markers after exercise (p>0.05). ConclusionCE and IE cause isoleucine, l-carnitine, and acylcarnitine levels to decrease, whereas ketone bodies were increased, thus indicating the energy metabolism shift from carbohydrates to amino acid utilization and lipid metabolism in COPD. Compared with CE, IE produces significant changes in more metabolomics in terms of carbohydrates, lipids, amino acids, nucleotides, and vitamins. Acute CE and IE alter circulating GPx levels in COPD.

Endogenous n-3 PUFAs Improve Non-Alcoholic Fatty Liver Disease through FFAR4-Mediated Gut-Liver Crosstalk.

The gut-liver axis plays a key role in the development and progression of non-alcoholic fatty liver disease (NAFLD). Due to the complexity and incomplete understanding of the cross-talk between the gut and liver, effective therapeutic targets are largely unknown. Free fatty acid receptors (FFARs) may bridge the cross-talk between the gut and liver. FFAR4 has received considerable attention due to its important role in lipid metabolism. However, the role of FFAR4 in this cross talk in NAFLD remains unclear. In this study, mice with high endogenous n-3 PUFAs but FFAR4 deficiency were generated by crossbreeding Fat-1 and FFAR4 knockout mice. FFAR4 deficiency blocked the protective effects of high endogenous n-3 PUFAs on intestinal barrier dysfunction and hepatic steatosis. In addition, FFAR4 deficiency decreased gut microbiota diversity and enriched Rikenella, Anaerotruncus, and Enterococcus, and reduced Dubosiella, Ruminococcaceae UCG-010, Ruminococcaceae UCG-014, Coriobacteriaceae UCG-002, Faecalibaculum, Ruminococcaceae UCG-009, and Akkermansia. Notably, FFAR4 deficiency co-regulated pantothenic acid and CoA biosynthesis, β-alanine metabolism, and sphingolipid metabolism pathways in the gut and liver, potentially associated with the aggravation of NAFLD. Together, the beneficial effects of n-3 PUFAs on the gut and liver were mediated by FFAR4, providing insights on the role of FFAR4 in the treatment of NAFLD through the gut-liver axis.

Development of a certified reference material for the analysis of vitamins in multivitamin tablets

Multivitamin tablet certified reference material (CRM, 108-10-019) was developed for the analysis of seven water-soluble vitamins, including thiamine, riboflavin, nicotinamide, pantothenic acid, pyridoxine, biotin, and folic acid. The CRM was prepared in powder form by grinding multivitamin tablets and then mixing, sieving, and bottling the powder. For the certification of each water-soluble vitamin, the isotope dilution mass spectrometry based on the liquid chromatography was applied. The methods for each analyte were validated by confirming the repeatability and reproducibility and by comparing with other CRMs. The property values and uncertainties for the vitamins were determined with 10 units from sample stored at − 20 °C. The homogeneity of each certified component was also examined in the range of 0.48–2.2%. All certified values for the seven water-soluble vitamins were stable for 3 or 6 years after the initial certification under storage conditions at − 20 °C. For fat-soluble vitamins, including retinol, α-tocopherol, cholecalciferol, and phylloquinone, two expert laboratories participated in analyses based on official methods, and the mean values of the reported results were assigned as reference values. The multivitamin tablet CRM (108-10-019) will be useful for validating analytical methods and for ensuring the quality of results for vitamin analysis in multivitamin tablets or similar products.

Agronomic Performance and Phytochemical Profile of Lettuce Grown in Anaerobic Dairy Digestate

Anaerobic liquid dairy digestate is a by-product of dairy waste anaerobic digestion from dairy operations and is associated with environmental risks if not handled properly, particularly nutrient leaching losses, water contaminations, and greenhouse gas emissions. We tested the applications of anerobic digestate (AD) as a biofertilizer and water source in greenhouse vegetable production to integrate food production and industry waste management for sustainable environments. We used a deep water culture system to assess the effects of AD effluent alone, inorganic nutrient solution (NS), and a combination of AD and NS on the growth, yield, and phytonutrient profile and heavy metal contamination assessment of hydroponically produced lettuce. Lettuce produced in AD had a lower leaf area, total chlorophyll content, and fresh biomass; however, it displayed significantly higher chicoric acid (200%), chlorogenic acid (67%), luteolin (800%), quercetin-3-O-β-d-glucuronide (378%), quercetin-3-glucoside (200%), quercetin-3-O-(6″-O-malonyl)-β-D-glucoside (1077%), folate (248%), pantothenic acid (200%), total phenolics (111%), total antioxidants (44%), and soluble sugars (253%) compared to control (inorganic feed). The AD-produced lettuce also showed significantly lower heavy metal bioaccumulation risks associated with the human consumption. Based on various results, we may conclude that AD utilization in hydroponics can offer a sustainable solution to harvest a better lettuce yield, higher phytonutrients, and environmental benefits.

Serum organic acid metabolites can be used as potential biomarkers to identify prostatitis, benign prostatic hyperplasia, and prostate cancer.

Noninvasive methods for the early identify diagnosis of prostatitis, benign prostatic hyperplasia (BPH), and prostate cancer (PCa) are current clinical challenges. The serum metabolites of 20 healthy individuals and patients with prostatitis, BPH, or PCa were identified using untargeted liquid chromatography-mass spectrometry (LC-MS). In addition, targeted LC-MS was used to verify the organic acid metabolites in the serum of a validation cohort. Organic acid metabolites had good sensitivity and specificity in differentiating prostatitis, BPH, and PCa. Three diagnostic models identified patients with PROSTATITIS: phenyllactic acid (area under the curve [AUC]=0.773), pyroglutamic acid (AUC=0.725), and pantothenic acid (AUC=0.721). Three diagnostic models identified BPH: citric acid (AUC=0.859), malic acid (AUC=0.820), and D-glucuronic acid (AUC=0.810). Four diagnostic models identified PCa: 3-hydroxy-3-methylglutaric acid (AUC=0.804), citric acid (AUC=0.918), malic acid (AUC=0.862), and phenyllactic acid (AUC=0.713). Two diagnostic models distinguished BPH from PCa: phenyllactic acid (AUC=0.769) and pyroglutamic acid (AUC=0.761). Three diagnostic models distinguished benign BPH from PROSTATITIS: citric acid (AUC=0.842), ethylmalonic acid (AUC=0.814), and hippuric acid (AUC=0.733). Six diagnostic models distinguished BPH from prostatitis: citric acid (AUC=0.926), pyroglutamic acid (AUC=0.864), phenyllactic acid (AUC=0.850), ethylmalonic acid (AUC=0.843), 3-hydroxy-3-methylglutaric acid (AUC=0.817), and hippuric acid (AUC=0.791). Three diagnostic models distinguished PCa patients with PROSTATITISA < 4.0 ng/mL from those with PSA > 4.0 ng/mL: 5-hydromethyl-2-furoic acid (AUC=0.749), ethylmalonic acid (AUC=0.750), and pyroglutamic acid (AUC=0.929). Conclusions: These results suggest that serum organic acid metabolites can be used as biomarkers to differentiate prostatitis, BPH, and PCa.

Targeted metabolomics characterizes metabolite occurrence and variability in stable freshwater mussel populations.

Freshwater mussels (order Unionida) play a key role in freshwater systems as ecosystem engineers and indicators of aquatic ecosystem health. The fauna is globally imperilled due to a diversity of suspected factors; however, causes for many population declines and mortality events remain unconfirmed due partly to limited health assessment tools. Mussel-monitoring activities often rely on population-level measurements, such as abundance and age structure, which reflect delayed responses to environmental conditions. Measures of organismal health would enable preemptive detection of declining condition before population-level effects manifest. Metabolomic analysis can identify shifts in biochemical pathways in response to stressors and changing environmental conditions; however, interpretation of the results requires information on inherent variability of metabolite concentrations in mussel populations. We targeted metabolites in the haemolymph of two common mussels, Lampsilis cardium and Lampsilis siliquoidea, from three Indiana streams (USA) using ultra-high-performance liquid chromatography combined with quadrupole time-of-flight mass spectroscopy. The influence of species, stream and sex on metabolite variability was examined with distance-based redundancy analysis. Metabolite variability was most influenced by species, followed by site and sex. Inter- and intraspecies metabolite variability among sexes was less distinct than differences among locations. We further categorized metabolites by occurrence and variability in mussel populations. Metabolites with high occurrence (Categories 1 and 2) included those indicative of energy status (catabolism versus anabolism; arginine, proline, carnitine, nicotinic acid, pantothenic acid), oxidative stress (proline, glutamine, glutamate) and protein metabolism (thymidine, cytidine, inosine). Metabolites with lower occurrence (Category 3) are constituents of assorted metabolic pathways and can be important biomarkers with additional temporal sampling to characterize their variability. These data provide a reference for future temporal (before/after) monitoring and for studies of stressor-metabolite linkages in freshwater mussels.

Evaluating the recovery of bioactive compounds and antioxidant activity of unripe red grape liquid extracts obtained by maceration

This paper evaluates the results of extracting bioactive compounds using modern maceration technology, which transforms food products into a liquid extract, based on the example of unripe grapes and its by-product called verjuice. During the extraction process, the indicators of niacin and pyridoxine remained stable, while pantothenic acid and choline increased significantly. During the extraction, the extraction system produced 712 kg of liquid extract, which was characterized by a high content of phenolic compounds and water-soluble vitamins, as well as a high antioxidant activity compared to the measurement obtained for green juice from unripe grapes. Since the estimated liquid extract yield from 950 kg of crushed immature grapes is 847 kg, the actual liquid extract yield was about 80%. The advantages of the maceration extraction method included avoiding the use of solvents or preservatives, low temperature treatment and little oxidative damage. Besides, it enabled enhancing the extraction yield due to introduction of certain post-extraction steps.

Effect of a 1-Year Nutritional Blend Supplementation on Plasma p-tau181 and GFAP Levels among Community-Dwelling Older Adults: A Secondary Analysis of the Nolan Trial.

Observational studies and some randomized controlled trials have suggested that nutritional supplementation could be a possible intervention pathway to prevent cognitive decline and Alzheimer's disease (AD). As measuring amyloid-β and tau pathophysiology by positron emission tomography (PET) or cerebrospinal fluid (CSF) analyses may be perceived as complex, plasma versions of such biomarkers have emerged as more accessible alternatives with comparable capacity of predicting cognitive impairment. This study aimed to evaluate the effect of a 1-year intervention with a nutritional blend on plasma p-tau181 and glial fibrillary acidic protein (GFAP) levels in community-dwelling older adults. Effects were further assessed in exploratory analyses within sub-cohorts stratified according to p-tau status (with the third tertile considered as high: ≥15.1 pg/ mL) and to apolipoprotein E (APOE) ε4 allele status. A total of 289 participants ≥70 years (56.4% female, mean age 78.1 years, SD=4.7) of the randomized, double-blind, multicenter, placebo-controlled Nolan trial had their plasma p-tau181 assessed, and daily took either a nutritional blend (composed of thiamin, riboflavin, niacin, pantothenic acid, pyridoxine, biotin, folic acid, cobalamin, vitamin E, vitamin C, vitamin D, choline, selenium, citrulline, eicosapentaenoic acid - EPA, and docosahexaenoic acid - DHA) or placebo for 1 year. After 1-year, both groups presented a significant increase in plasma p-tau181 and GFAP values, with no effect of the intervention (p-tau181 between-group difference: 0.27pg/mL, 95%CI: -0.95, 1.48; p=0.665; GFAP between-group difference: -3.28 pg/mL, 95%CI: -17.25, 10.69; p=0.644). P-tau-and APOE ε4-stratified analyses provided similar findings. In community-dwelling older adults, we observed an increase in plasma p-tau181 and GFAP levels that was not different between the supplementation groups after one year.

Industry survey of added vitamins and trace minerals in U.S. swine diets.

From November 2021 to February 2022, 37 swine nutritionists representing 29 production systems and 8 nutrition supplier companies in the United States were surveyed about added vitamin and trace mineral concentrations in swine diets. Respondents were asked to provide vitamin premix and trace mineral concentrations, inclusion rates, and weight ranges associated with feeding phases. Survey participants represented 4.38 million sows, or 72% of the U.S. industry. Data were compiled into three nursery phases (phase 1, weaning to 7 kg; phase 2, 7 to 11 kg; and phase 3, 11 to 23 kg), three finishing phases (23 to 55 kg; 55 to 100 kg; 100 kg to market), gilt development, gestation, lactation, and boar. Within each dietary phase, the vitamins and trace minerals of interest included: vitamins A, D, E, and K, thiamin, riboflavin, niacin, pantothenic acid, pyridoxine, biotin, folic acid, vitamin B12, choline, vitamin C, carnitine, copper, iodine, iron, manganese, selenium, zinc, cobalt, and chromium. Descriptive statistics used included: average, weighted average (determined by the total number of sows represented), median, minimum, maximum, 25th percentile (lowest quartile), and 75th percentile (highest quartile). In addition, all average supplementation rates for vitamins and trace minerals within each phase of production were compared to the requirement estimates reported in the NRC (2012). Nutritionists generally supplemented vitamins and trace minerals well above the NRC (2012) requirement estimates. However, great variation among respondents was observed in all vitamins and trace minerals, particularly in the fat-soluble vitamins. Also, the use of alternative sources of vitamin D [25(OH)D3], E (natural, d-alpha-tocopherol), and organic or chelated minerals like copper, manganese, selenium, and zinc were being used by approximately 40% of the respondents, primarily in breeding herd and nursery diets. Understanding current supplementation practices may help develop research trials to test different vitamin and trace mineral inclusions and provide an industry benchmark of vitamin and trace mineral usage.

Evaluation of a ruminally protected blend of pantothenic acid, pyridoxine, folic acid, biotin, and vitamin B12 on finishing steer growth performance, efficiency of dietary net energy utilization, carcass trait responses, and liver abscess prevalence and severity.

The objective of this study was to determine the influence that a ruminally-protected B-vitamin (RPBV) blend (containing vitamin B5, B6, B7, B9, and B12) had on growth performance, efficiency of dietary net energy utilization, carcass trait responses, and liver abscess severity and prevalence in beef steers fed a finishing diet. Steers (n = 246; initial shrunk body weight [BW] = 411 ± 25.8 kg) from two different sources, were used in a 126-d RCBD experiment. Within 48 h after arrival, steers were individually weighed and allotted to 1 of 24 pens (n = 8 to 12 steers; 8 pens per treatment) and randomly assigned to 1 of 3 treatments: (1) No RPBV; (2) RPBV1 at 1 g/steer d-1; 3) RPBV2 at 2 g/steer d-1. During the first 14 d, cattle received two transition diets with increasing concentrate. From days 15 to 126, cattle were fed the final diet containing 53% dry-rolled corn; 23% corn silage; 20% MDGS; and 4% suspended supplement. On the first 28 d, steers of RPBV1 had a greater average daily gain (ADG) and better feed conversion (G:F), both by 9% (quadratic effect, P ≤ 0.02). However, cumulatively, no differences (P ≥ 0.13) among treatments were found for dry-matter intake (DMI), live final BW, ADG, or G:F. Carcass-adjusted final BW, ADG, and G:F were not influenced by treatment (P ≥ 0.59). Additionally, carcass weight, dressing percentage, marbling score, kidney-pelvic-heart fat, or BW at 28% empty body fat did not differ among treatments (P ≥ 0.11). Ribeye area (REA) was altered (quadratic effect, P = 0.02) by treatment; steers from RPBV1 had decreased REA compared to others. Additionally, calculated yield grade (YG) and calculated retail yield (RY) were altered (quadratic effect, P ≤ 0.01) by treatment; steers from RPBV1 had increased YG and decreased RY compared to others. Estimated empty body fatness tended (P = 0.06) to be greater from steers-fed RPBV compared to control. Overall USDA YG distribution was altered by dietary treatment (P = 0.01). The proportions of YG1 and YG5 carcasses were unaffected by treatment, but there was a shift in the proportion of carcasses that graded YG2, YG3, and YG4 among treatments. Distribution of USDA Quality Grade was not altered by treatment (P = 0.53). No treatment differences in liver abscess incidence or severity were observed (P = 0.13). The use of RPBV altered carcass muscularity and rib fat accumulation affecting the overall YG distribution. However, RPBV did not appreciably influence any cumulative growth performance measures or liver abscess outcome.

A Promising Antibacterial Wound Dressing Made of Electrospun Poly (Glycerol Sebacate) (PGS)/Gelatin with Local Delivery of Ascorbic Acid and Pantothenic Acid

A Promising Antibacterial Wound Dressing Made of Electrospun Poly (Glycerol Sebacate) (PGS)/Gelatin with Local Delivery of Ascorbic Acid and Pantothenic Acid

Evidence for a Conserved Function of Eukaryotic Pantothenate Kinases in the Regulation of Mitochondrial Homeostasis and Oxidative Stress.

Human PANK1, PANK2, and PANK3 genes encode several pantothenate kinase isoforms that catalyze the phosphorylation of vitamin B5 (pantothenic acid) to phosphopantothenate, a critical step in the biosynthesis of the major cellular cofactor, Coenzyme A (CoA). Mutations in the PANK2 gene, which encodes the mitochondrial pantothenate kinase (PanK) isoform, have been linked to pantothenate-kinase associated neurodegeneration (PKAN), a debilitating and often fatal progressive neurodegeneration of children and young adults. While the biochemical properties of these enzymes have been well-characterized in vitro, their expression in a model organism such as yeast in order to probe their function under cellular conditions have never been achieved. Here we used three yeast mutants carrying missense mutations in the yeast PanK gene, CAB1, which are associated with defective growth at high temperature and iron, mitochondrial dysfunction, increased iron content, and oxidative stress, to assess the cellular function of human PANK genes and functional conservation of the CoA-controlled processes between humans and yeast. Overexpression of human PANK1 and PANK3 in these mutants restored normal cellular activity whereas complementation with PANK2 was partial and could only be achieved with an isoform, PanK2mtmΔ, lacking the mitochondrial transit peptide. These data, which demonstrate functional conservation of PanK activity between humans and yeast, set the stage for the use of yeast as a model system to investigate the impact of PKAN-associated mutations on the metabolic pathways altered in this disease.

Characterization and temporal dynamics of the intra- and extracellular environments of Lactiplantibacillus plantarum using multi-platform metabolomics

The characterization and temporal changes of intracellular and extracellular metabolome of Lactiplantibacillus plantarum under different growth times were studied using multi-platform metabolomics. A multi-platform approach consisting of both non-targeted profiling and targeted quantification has identified a total of 168 metabolites among which, 41 and 110 in the intra- and extracellular metabolomes were determined to be significantly different. We constructed an expanded metabolic pathway map that combines the amino acid, fatty acid, carbohydrate, purine, and pyrimidine pathways. Notably, we observed the dramatic increase of bioactive compound during growth such as indoles, phenylpropanoid, and hydroxy fatty acids produced by amino acid and fatty acid metabolism in the extracellular metabolome. Several B vitamins (pantothenic acid in intracellular metabolome and nicotinic acid, pyridoxine, and pyridoxal in extracellular metabolome) significantly increased in specific growth phase. In conclusion, this study has demonstrated the usefulness of a multi-platform approach in identifying a large pool of metabolites which, thus, has led, to a more robust understanding of the metabolic processes involved in the growth of L. plantarum HAC01. Furthermore, the change of discriminant metabolites with integrated metabolic pathway were revealed, which can be useful in identifying candidates for compound markers and postbiotic components for commercialization of probiotics formulation.

Detection of new phytochemical compounds from Vassobia breviflora (Sendtn.) Hunz: antioxidant, cytotoxic, and antibacterial activity of the hexane extract

ABSTRACT Vassobia breviflora (Sendtn.) Hunz is a plant of the Solanaceae family from South America and there are no apparent studies reported on the biological activity of the hexane extract. The aim of this investigation was to conduct phytochemical analyses using ESI-TOF-MS, while antioxidant activities were evaluated by the following methods 1,1-diphenyl-2-picrylhydrazyl (DPPH) 2,2“-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical capture (ABTS), ferric reducing antioxidant power (FRAP), total antioxidant capacity (TAC), and total oxidant status (TOS). Antimicrobial activities were performed by determining the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiofilm formed. Cytotoxicity was measured by MTT and dsDNA PicoGreen tests, beyond the production of reactive oxygen species (ROS) determined by Dichlorodihydrofluorescein diacetate (DCFH-DA). The hexane extract showed the presence of 5 (choline, pantothenic acid, calystegine B, lanciphodylactone I, and 15”-cis-zeaxanthin) compounds detected. V. breviflora extract demonstrated reliable results utilizing different antioxidant methods. In antibacterial activity, V. breviflora extract exhibited inhibitory, bactericidal, and antibiofilm action in biofilm-forming bacteria. The hexane extract exhibited cytotoxicity against melanoma, lung cancer, glioblastoma, leukemia, uterine colon, and hepatocarcinoma tumor cells. In addition, all tested strains resulted in increased production of ROS. This plant extract may be considered in future as an alternative for development of new therapeutic options aimed at the treatment of diverse pathologies.

Sodium Multivitamin Transporter-Targeted Fluorochrome Facilitates Enhanced Metabolic Evaluation of Tumors Through Coenzyme-R Dependent Intracellular Signaling Pathways.

Intraoperative molecular imaging (IMI)-guided resections have been shown to improve oncologic outcomes for patients undergoing surgery for solid malignancies. The technology utilizes fluorescent tracers targeting cancer cells without the use of any ionizing radiation. However, currently available targeted IMI tracers are effective only for tumors with a highly specific receptor expression profile, and there is an unmet need for IMI tracers to label a broader range of tumor types. Here, we describe the development and testing of a novel tracer (CR)-S0456) targeted to the sodium multivitamin transporter (SMVT). Preclinical models of fibrosarcoma (HT-1080), lung (A549), breast (4T1), and renal cancers (HEK-293T) in vitro and in vivo were used for assessment of (CR)-S0456 specific tumor labeling via sodium-mediated SMVT uptake in dipotassium phosphate or choline chloride-containing media buffer. Additionally, pharmacologic inhibition of multiple intracellular coenzyme-R obligate signaling pathways, including holocarboxylase synthetase (sulconazole nitrate), PI3K/AKT/mTOR (omipalisib), and calmodulin-dependent phosphatase (calmidazolium), were investigated to assess (CR)-S0456 uptake kinetics. Human fibrosarcoma-bearing xenografts in athymic nude mice were used for tumor and metabolic-specific labeling. Novel NIR needle confocal laser endomicroscopic (nCLE) intratumoral sampling was performed to demonstrate single-cell specific labeling by CR-S0456. CR-S0456 localization in vitro correlated with highly proliferative cell lines (MTT) and doubling time (p < 0.05) with the highest microscopic fluorescence detected in aggressive human fibrosarcomas (HT-1080). Coenzyme-R-specific localization was demonstrated to be SMVT-specific after competitive inhibition of internal localization with excess administration of pantothenic acid. Inhibiting the activity of SMVT by affecting sodium ion hemostasis prevented the complete uptake of CR-S0456. In vivo validation demonstrated (CR)-S0456 localization to xenograft models with accurate identification of primary tumors as well as margin assessment down to 1 mm3 tumor volume. Systemic treatment of xenograft-bearing mice with a dual PI3K/mTOR inhibitor suppressed intratumoral cell signaling and (CR)-S0456 uptake via a reduction in SMVT expression. Novel analysis of in vivo intratumoral cytologic fluorescence using near-infrared confocal laser endomicroscopy demonstrated the absence of coenzyme-R-mediated NIR fluorescence but not fibroblast activation protein (FAP)-conjugated fluorochrome, indicating specific intracellular inhibition of coenzyme-R obligate pathways. These findings suggest that a SMVT-targeted NIR contrast agent can be a suitable tracer for imaging a wide range of malignancies as well as evaluating metabolic response to systemic therapies, similar to PET imaging with immune checkpoint inhibitors.

Phenotypic characterization and genome analysis reveal the probiotic potential of a banyan endophyte Bacillus velezensis K1.

The current study aimed to scrutinize the probiotic traits and safety aspects of Bacillus velezensis K1 through experimental and supporting genome studies. The seven cultures previously isolated from the aerial roots of Ficus benghalensis were initially screened for their antibacterial activity as well as acid and bile tolerance. The isolate K1 was found to be the most potent and was further investigated for probiotic traits and safety. K1 showed tolerance to simulated digestive juices and 0.3% bile. It showed notable aggregation, cell surface hydrophobicity, and adherence to HT-29 cells. K1 significantly prevented the adhesion of E. coli O157: H7 and S. enterica ATCC 13076 to HT-29 in cell culture assays. K1 could hydrolyze phytate and complex polysaccharides. The genes related to stress tolerance, adhesion, antimicrobial activity, and production of vitamins, viz. thiamine, riboflavin, pyridoxine, pantothenic acid, folate, and biotin were annotated in the K1 genome. K1 was found to be non-hemolytic, noncytotoxic, as well as susceptible to antibiotics. No virulence or toxin-encoding genes were identified in its genome. B. velezensis K1 is a prospective probiotic with the ability to tolerate gastrointestinal stress, adhere to intestinal surfaces, and inhibit enteropathogens.

In Vitro, In Silico and Network Pharmacology Mechanistic Approach to Investigate the α-Glucosidase Inhibitors Identified by Q-ToF-LCMS from Phaleria macrocarpa Fruit Subcritical CO2 Extract.

The fruit of Phaleria macrocarpa have been traditionally used as an antidiabetic remedy in Malaysia and neighbouring countries. Despite its potential for diabetes treatment, no scientific study has ever been conducted to predict the inhibitor interaction of the protein α-glucosidase identified in an extract prepared with a non-conventional extraction technique. Hence, the major aim of this research was to evaluate the in vitro antioxidant, the α-glucosidase inhibitors, and the molecular dynamic simulations of the α-glucosidase inhibitors identified by Quadrupole Time-of-Flight Liquid Chromatography Mass Spectrometry (Q-ToF-LCMS) analysis. Initially, dry fruit were processed using non-conventional and conventional extraction methods to obtain subcritical carbon dioxide extracts (SCE-1 and SCE-2) and heating under reflux extract (HRE), respectively. Subsequently, all extracts were evaluated for their in vitro antioxidative and α-glucosidase inhibitory potentials. Subsequently, the most bioactive extract (SCE-2) was subjected to Q-ToF-LCMS analysis to confirm the presence of α-glucosidase inhibitors, which were then analysed through molecular dynamic simulations and network pharmacology approaches to confirm their possible mechanism of action. The highest inhibitory effects of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and α-glucosidase on SCE-2 was found as 75.36 ± 0.82% and 81.79 ± 0.82%, respectively, compared to the SCE-1 and HRE samples. The Q-ToF-LCMS analysis tentatively identified 14 potent α-glucosidase inhibitors. Finally, five identified compounds, viz., lupenone, swertianolin, m-coumaric acid, pantothenic acid, and 8-C-glucopyranosyleriodictylol displayed significant stability, compactness, stronger protein-ligand interaction up to 100 ns further confirming their potential as α-glucosidase inhibitors. Consequently, it was concluded that the SCE-2 possesses a strong α-glucosidase inhibitory effect due to the presence of these compounds. The findings of this study might prove useful to develop these compounds as alternative safe α-glucosidase inhibitors to manage diabetes more effectively.

Radotinib attenuates TGFβ -mediated pulmonary fibrosis in vitro and in vivo: exploring the potential of drug repurposing

BackgroundTyrosine kinase (TK) plays a crucial role in the pathogenesis of idiopathic pulmonary fibrosis. Here, we aimed to investigate whether radotinib (Rb) could inhibit pulmonary fibrosis by inhibiting TK in vitro and in vivo.MethodsThe antifibrotic effects of Rb in transforming growth factor-β (TGF-β)1-stimulated A549 cells were determined using real-time polymerase chain reaction, western blotting, and immunocytochemistry assays. Rb inhibition of bleomycin-induced lung fibrosis in Sprague Dawley (SD) rats was determined by histopathological and​ immunohistochemical analyses. Rb-interfering metabolites were analyzed using LC-MS/MS.ResultsRb concentrations of up to 1000 nM did not affect the viability of A549 cells, but Rb (30 nM) significantly reduced expression of TGF-β1 (10 ng/mL)-induced ECM factors, such as Snail, Twist, and F-actin. Rb also regulated TGF-β1-overexpressed signal cascades, such as fibronectin and α-smooth muscle actin. Furthermore, Rb attenuated the phosphorylation of Smad2 and phosphorylation of kinases, such as, extracellular signal-regulated kinase, and protein kinase B. In the inhibitory test against bleomycin (5 mg/kg)-induced lung fibrosis, the Rb (30 mg/kg/daily)-treated group showed a half-pulmonary fibrosis region compared to the positive control group. In addition, Rb significantly reduced collagen type I and fibronectin expression in the bleomycin-induced fibrotic region of SD rats. Further, the identified metabolite pantothenic acid was not altered by Rb.ConclusionTaken together, these results indicate that Rb inhibits TGF-β1-induced pulmonary fibrosis both in vitro and in vivo. These findings suggest that Rb may be an effective treatment for pulmonary fibrosis-related disorders and idiopathic pulmonary fibrosis.

Identification of nutritional components in unripe and ripe Docynia delavayi (Franch.) Schneid fruit by widely targeted metabolomics.

Docynia delavayi (Franch.) Schneid is an evergreen tree with multiple benefits and high development and utilization value. The fruit is consumed as fresh and dry fruit, juices, and other products. However, it is unknown the chemical changes that occur upon fruit maturation. The metabolite content of unripe and ripe fruit was examined using UPLC-MS/MS technology based on a broadly targeted metabolome. We identified 477 metabolites, of which 130 differed between ripe and unripe fruit. These compounds are primarily involved in the biosynthesis of secondary metabolites, such as pantothenic acid, flavonoids, and amino acids. Moreover, in ripe fruit, there are 94 metabolites that are upregulated, particularly flavonoids and terpenoids. In comparison, compounds associated with sour flavors (amino acids, phenolic acids, organic acids) are down-regulated. Remarkably, these metabolites have a strong relationship with the medicinal properties of D. delavayi. This study provides a global perspective of the D. delavayi fruit metabolome and a comprehensive analysis of metabolomic variations during fruit development, thereby increasing the knowledge of the metabolic basis of important fruit quality traits in D. delavayi fruit.