bluuebunny/crossref_metadata_2025_split huggingface dataset:
Vitamin D Reduces Hepcidin Concentrations Independent of Inflammatory Cytokines in Healthy Adults: A Pilot, Randomized, Double‐Blind, Placebo‐Controlled Trial of High‐Dose Vitamin D3
Authors: Smith Ellen , Alvarez Jessica , Kearns Malcolm , Hao Li , Sloan John , Konrad Robert , Ziegler Thomas , Zughaier Susu , Tangpricha Vin 
Published: November 2023

ObjectiveDisturbances in iron recycling may result from elevations in pro‐inflammatory cytokines and hepcidin, the major iron‐regulatory hormone. In vitro studies suggest that vitamin D may reduce hepcidin and pro‐inflammatory cytokines, and vitamin D deficiency is associated with increased odds of anemia of inflammation. However, data from clinical trials assessing the vitamin D‐mediated effects on iron recycling in healthy individuals are lacking. Our objectives were to: 1) examine the effect of high‐dose vitamin D on plasma hepcidin, inflammatory cytokine, and ferritin concentrations in healthy adults; and 2) determine whether any observed changes in hepcidin were concomitant with or independent of changes in pro‐inflammatory cytokines.MethodsThis was a pilot, double‐blind, placebo‐controlled trial in healthy adults (N=28) randomized to receive a one‐time oral dose of 250,000 IU of vitamin D3 or placebo. Between‐and within‐group differences in plasma hepcidin, pro‐inflammatory cytokine [interleukin (IL)‐1β, IL‐6, IL‐8, monocyte chemoattractant protein‐1 (MCP‐1)], and ferritin concentrations at baseline and 1 week were determined using two‐sample and paired t‐tests, respectively.ResultsAt baseline, plasma 25‐hydroxyvitamin D [25(OH)D], hepcidin, pro‐inflammatory cytokine, and ferritin concentrations did not differ between the two randomized groups; 75% of subjects had plasma 25(OH)D concentrations < 20 ng/mL. After 1 week, plasma hepcidin concentrations had decreased significantly from baseline by 73% in those who received vitamin D3 (geometric mean ratio [GMR]: 0.27 (95% CI: 0.11, 0.62); P=0.005) compared to no significant change with placebo (GMR: 0.73 (95% CI: 0.49, 1.09); P=0.11). Plasma cytokine and ferritin concentrations did not change significantly in either group.ConclusionHigh‐dose vitamin D3 significantly reduced plasma hepcidin concentrations in healthy adults 1 week post‐dosing, while no change was observed in pro‐inflammatory cytokine or ferritin concentrations. These findings suggest that vitamin D may have a role in regulating iron recycling by acting directly on hepcidin, independent of changes in inflammatory markers; however, down‐stream effects on markers of iron status in the absence of inflammatory conditions remain unclear. Longer follow‐up and evaluation of additional markers of iron status may be required.Support or Funding InformationT32 DK007734 (EMS), K01 DK102851 (JAA), K24 DK096574 (TRZ), UL1 TR000454 (TRZ, VT)
Effects of Calorie Restriction on Microglial Activation in a murine model
Authors: Quijano Uriel , Contreras Irazú , Estrada José A. 
Published: November 2023

Calorie restriction has been shown to have beneficial physiological effects in different organisms, delaying the onset of chronic non‐communicable diseases, increasing maximum lifespan, as well as reducing excess body weight and the risk of diseases associated with aging. Recent studies have shown that calorie restriction inhibits the morphological change that is associated with activated microglia, but so far, no data are available to describe if caloric restriction somehow alters the effector functions of microglia.To determine the effect of calorie restriction on microglia, we analyzed purified mononuclear cells from the brain and spinal cord of 8 weeks old BALB/c mice, divided into three experimental groups: ad libitum, calorie restriction (30% reduction in food intake) and hiper‐energetic (supplemented with sucrose in drinking water), exposed to this diet for 4 weeks. After treatment, brain and spinal cord samples were obtained and microglial cells were purified by Percoll density gradients centrifugation. Purified cells were stained with antibodies for MHC‐I, MHC‐II, CD80 and CD86, as well as cell surface markers CD45 and CD11b and anlyzed by flow cytometry.Our results show no apparent statistically significant difference in the frequency of MHC+ cells in the experimental groups compared with the control group. The frequency of CD80 and CD86 single positive cells both appear to be increased in the CR group, compared to the control, although the frequency of cells expressing these markers is very low in all groups. We are analyzing the mean fluorescence intensity of all markers to determine if there is a difference in their expression among groups.
In vitro Comparison of Anti‐Inflammatory Activity of Green Tea Catechins
Authors: Fechtner Sabrina , Ahmed Salahuddin 
Published: November 2023

Background/PurposeGreen tea contains considerable amounts of epigallocatechin‐3‐gallate (EGCG), epigallocatechin (EGC), and epicatechin (EC). EGCG has already been shown to exhibit anti‐inflammatory activity. Pro‐inflammatory cytokine interleukin‐1β (IL‐1β) produced by monocytes and macrophages plays an important role in rheumatoid arthritis (RA). In this study, we compared the efficacy of different catechins in reducing the inflammatory response to IL‐1β in human RA synovial fibroblasts (RA‐FLS).MethodsThe effect of EGCG, EGC, and EC on RA‐FLS viability was tested using MTT assay. RA‐FLS were pre‐treated with different concentration EGCG, EGC, and EC (5–20 μM) overnight then stimulated with IL‐1β (10 ng/ml) for 24 hours. Condition media was used to quantify IL‐6 and IL‐8 production by ELISA, and MMP‐2 activity by gelatin zymography. The cellular expression of cyclooxygenase‐2 (Cox‐2) and Cox‐1 was evaluated using Western blotting method.ResultsEGCG, EGC, and EC exhibit no toxicity in human RA‐FLS (n=3). Among the tested catechins, EGCG was the most effective catechin at inhibiting IL‐6 and IL‐8 production by 59% and 57%, respectively (n=3; p<0.05). Western blot analysis showed that EGCG selectively inhibits Cox‐2 expression by 86% after IL‐1β stimulation (n=3; p<0.05) and had no significant effect on IL‐1β‐induced Cox‐1 expression. However, EGC not only inhibited IL‐1β‐induced Cox‐2 expression by 58%, it also reduced IL‐1β‐induced Cox‐1 expression by 20% (n=3; p<0.05). Interestingly, our gelatin zymography results showed that EGC was as effective as EGCG in reducing IL‐1β‐induced MMP‐2 activity by 85% in human RA‐FLS (n=3; p<0.05). In all the proposed assays, EC showed no anti‐inflammatory properties.ConclusionThe results of the study suggest that green tea catechins differentially interfere with IL‐1β signaling pathways to regulate IL‐6 and IL‐8 production, and Cox‐2, Cox‐1, and MMP‐2 expression in human RA‐FLS. Among the catechins tested, EGCG was a more potent anti‐inflammatory compound than EGC and EC in human RA‐FLS. EGCG and EGC display moderate anti‐inflammatory properties while EC displays minimal anti‐inflammatory properties. Together, these results suggest that the gallate moiety is essential to EGCG and EGC inhibiting IL‐1β‐induced inflammatory mediators in RA.Support or Funding InformationThis study was supported by the Robbers Research Award (S.F.), NIH grant AR063104 (S.A.), the Arthritis Foundation Innovative Research Grant (S.A.), the start‐up funds from Washington State University (S.A.).
Cranberry and Recurrent Urinary Tract Infections (UTIs) in Generally Healthy Women: A Systematic Review and Meta‐analysis of Randomized Clinical Trials (RCTs)
Authors: Kern Hua , Liska DeAnn , Chung Mei 
Published: November 2023

The aim of the present meta‐analysis was to evaluate the efficacy of cranberry on the recurrence of uncomplicated urinary tract infections (UTIs) among otherwise healthy women. [Registered on International Prospective Register of Systematic Reviews: PROSPERO 2015:CRD42015024439] Two scientists independently conducted the literature search and extracted data, with discrepancies resolved via discussion with a third scientist. Meta‐analysis was performed using RevMan (version 5.3, Cochrane). Six studies (1320 subjects) were included in qualitative and quantitative synthesis. Cranberry reduced the risk of uncomplicated UTI recurrence among healthy women by 18% [Risk Ratio (RR) =0.82; 95% Confidence Interval (CI): 0.68–0.99; p=0.04] (I2=38%). When an outlier study was excluded from the analysis, cranberry further reduced the risk of UTI recurrence by 27%, compared to placebo/control (RR=0.73; 95% CI: 0.60–0.90; p=0.003) (I2=0%). Subgroup analysis indicated cranberry seemed to be more effective in women who were free of UTI at the start of the trial (RR=0.72; 95% CI: 0.55–0.94; p=0.02; n=3 studies) (I2=0%), or over a longer (12 months) follow‐up period (RR=0.61; 95% CI: 0.40–0.91; p=0.02; n=2 studies) (I2=0%), than those who had UTI at the study entry or were followed up for 6 months (p>0.10), respectively. Risk of bias assessment indicated half of the included trials did not provide the randomization information, two had high rates of loss to follow‐up, and two thirds were subject to other biases. Results of this meta‐analysis suggest cranberry can be a potential nutritive option for generally healthy women to prevent the recurrence of uncomplicated recurrent UTIs.Support or Funding InformationSupported by Ocean Spray Cranberries, Inc.
Quantitative Measurement of Plasma and Urinary Excretion of 3‐Hydroxyisovaleryl Carnitine by UPLC‐MS/MS
Authors: Wang Zhu 
Published: November 2023

ObjectiveAn increased plasma concentration of 3‐hydroxyisovaleryl carnitine (3HIA‐carnitine) results from impairment in the leucine catabolic pathway at the conversion of 3‐methyl‐crotonyl‐CoA to 3‐methylglutaconyl‐CoA which is caused by reduced activity of the biotin‐dependent enzyme 3‐methylcrotonyl‐CoA carboxylase. In order to validate plasma 3HIA‐carnitine as a novel biomarker of biotin deficiency rats models, an LC‐MS/MS method for the quantitation of 3HIA‐carnitine in plasma and urinary excretion has been established.Materials & MethodsPlasma and urinary excretion samples were centrifugated with high speed at 3000×g for 10 minutes, the MCX solid phase extraction column was used to filter and purify the liquid samples, the following procedures were elution, wash‐out, redissolve, detection. The chromatographic separation was achieved on a Waters Acquity UPLC BEH C18 column (1.7μm, 2.1×50mm), with a mobile phase composed of 0.1% formic acid in water‐methanol with gradient elution. Analyte quantification was performed in the positive electrospray ionization mode and multiple monitoring. The quantification and qualification were performed using the internal standard of 3‐Hydroxyisovaleryl carnitine, then the evaluation for accuracy and precision were followed.Main FindingsThe RSD of six parallel determination for the 3HIA‐carnitine in plasma and urine were 4.8%, 5.5%, respectively. The linear range of 3‐Hydroxyisovaleryl carnitine was 0.5~20ng/ml in the plasma 5~200ng/ml in urine, with both the linear correlation coefficient was above 0.99. Moreover in three standard levels, rate of recovery in plasma and urine were more than 102.5%, 79%. The detection and quantification limits for urine and plasma were 20ng/L, 50ng/L and 10ng/L, 33ng/L, respectively.Conclusion & RecommondationsThe sample preparation method is simple and fast, and the method can be used to analyze 3‐Hydroxyisovaleryl carnitine in plasma and urinary excretion efficiently and sensitively. If confirmed in deeper studies, 3HIA‐carnitine in plasma and urinary is likely to be an important indicator of biotin status in a variety of clinical circumstances especially for the high‐risk population like pregnant women.Support or Funding InformationYouth fund of national natural science fundation in China (81302428)
Metabolomic Profile of Serum 25‐Hydroxy‐Vitamin D
Authors: Nelson Shakira M , Anic Gabriella M , Panagiotou Orestis A , Mondul Alison , Weinstein Stephanie J , Albanes Demetrius 
Published: November 2023

The role of vitamin D in human health has been studied using a variety of approaches. In laboratory studies, the active form of vitamin D, 1,25(OH)2D3 has been shown to play a significant role in reducing autoimmune diseases, such as diabetes. In epidemiological studies, vitamin D has been associated with risk for some cancers but not others. For example, fairly consistent inverse (i.e., protective) associations with circulating 25‐hydroxy‐vitamin D (25(OH)D) have been observed for colorectal cancers, while direct (i.e., adverse) associations have been seen in prostate cancer studies. In order to gain a more comprehensive biological understanding of vitamin D associations with cancers and other health outcomes, we utilized a metabolomics approach to identify metabolites associated with serum 25(OH)D. Over 900 metabolites were measured from fasting blood samples collected at study entry between 1985 and 1988 in the Alpha‐Tocopherol, Beta‐Carotene Cancer Prevention (ATBC) Study, a cohort of Finnish male smokers ages 50–69 years. Baseline serum 25(OH)D was measured in 392 men using radioimmunoassay, chemiluminescence assays, and OCTEIA direct enzyme‐linked immunosorbent assay kits. Metabolites were profiled using ultrahigh performance liquid chromatography/mass spectroscopy and gas chromatography/mass spectroscopy (Metabolon, Inc., N.C.). Metabolite‐vitamin D associations were modeled using linear regression. A statistically significant (p<0.05) change in beta‐estimates per 1 standard deviation (SD) increment was seen for 117 metabolites, the majority of which were lipids, along with 26 amino acids, 10 xenobiotics, seven peptides, six cofactors or vitamins, and four carbohydrate metabolites. Six lipid metabolites were significant at the Bonferroni corrected threshold of p < 0.000053, including 3‐carboxy‐4‐methyl‐5‐propyl‐2‐furanpropanoic acid (CMPF) (beta=0.42/1 SD increment), stearoyl‐arachidonoyl‐glycerophosphoethanolamine (GPPE) (beta =−0.36/1 SD), stearoyl‐linoleoyl‐GPPE (beta = −0.35/1 SD), erucoyl sphingomyelin (beta = 0.32/1 SD), eicosapentaenoate (beta=0.24/1 SD) and docosahexaenoate (beta=0.21/1 SD). The identification of these metabolites may provide insights into the role of vitamin D in human health.Support or Funding InformationResearch support: Intramural Research Program of the National Cancer Institute, NIH.
LC3 Overexpression Perturbs Pancreatic Acinar Cell Homeostasis and Alters Pancreatitis Responses
Authors: Mareninova Olga , Jia Wenzhuo , Elperin Jason , Lotshaw Ethan , Pimienta Michael , Reicher Benjamin , Gukovsky Ilya , Gukovskaya Anna 
Published: November 2023

Background & AimsDespite accumulating evidence for important role of autophagy in pancreatitis, the underlying mechanisms are poorly understood. Conversion of the key autophagy‐related (Atg) protein Atg8/LC3 fromits cytosolic LC3‐I form to the lipidated LC3‐II form is a critical step in autophagosome formation. We used mice (over)expressing LC3 conjugated to green fluorescent protein (GFP‐LC3) to study the role of autophagy in pancreatic acinar cell physiology and pancreatitis.MethodsWild type and GFP‐LC3 mice were subjected to fasting (starvation is a strong physiological inducer of autophagy) or used in several dissimilar experimental models of acute pancreatitis, namely those induced by cerulein [an analog of cholecystokin in (CCK)‐8], L‐arginine, choline deficient/ethionine‐supplemented diet, or ethanol diet pluslow‐dose cerulein. We measured parameters of autophagy, acinar cell functions, e.g., amylase secretion, and pancreatitis responses.ResultsLC3 overexpression markedly increased the basal number of autophagic vacuoles in the pancreas; and stimulated starvation‐induced autophagy. In GFP‐LC3 mice subjected to pancreatitis, autophagy was impaired/inefficient, as manifest by accumulation of abnormally large vacuoles and p62‐positive protein aggregates. These effects were quantified by using immunoblot analysis and confocal microscopy of LC3 “puncta”. In addition, the retarded autophagic flux was measured in isolated pancreatic acinar cells by using lysosomalprotease inhibitors. One mechanism underlying these effects could be decreased LC3‐Ito LC3‐II conversion due to feedback down‐regulation of Atg5 and Atg7, which we found in pancreas of GFP‐LC3 mice (both Atg5 and Atg7 mediate the LC3‐I to LC3‐II conversion). In all pancreatitis models studied, LC3 over expression dramatically increased serum amylase levels, a hallmark response of acutepancreatitis. At the same time, pancreatic levels of amylase and trypsinogen were up‐regulated, possibly due to decreased autophagic protein degradation. Another potential underlying mechanism could be an increase in protein synthesis mediated by the mechanistic target of rapamycin (mTOR). Importantly, LC3 over expression worsened pancreatitis severity (e.g., necrosis); but it had no effect on CCK‐induced amylase secretion from acinar cells.ConclusionsResults indicate a critical role for LC3‐mediated autophagy in regulating exocrine pancreas homeostasis and pancreatitis responses. Although GFP‐LC3 mice are widely used to monitor autophagy, caution should be exercised as LC3 overexpression may perturb both physiologic and pathophysiologic pathways.Support or Funding InformationNIH/NIAAA R01AA019730, NIH/NIDDK P01DK098108
Neural Modulation of Liver Function to Regulate Lipid and Glucose Metabolism
Authors: Patel Yogi A , Butera Robert J. 
Published: November 2023

Mammalian blood glucose concentrations are maintained within well‐defined biological limits despite considerable fluctuations in the rate at which glucose is obtained from food and utilized by tissues. Of all homeostatic mechanisms, maintenance of blood glucose levels is finely regulated, and one in which the liver and the central nervous system play a prominent role. The liver can add or remove glucose from circulating blood in accordance with the demands of the body, which are transmitted by both hormonal and neural messaging. The hormonal messaging system is well understood, unlike the neural messaging system. Hormonal messaging is mediated by the pancreatic hormones insulin and glucagon, which have opposite effects on blood glucose levels. In contrast, the neural messaging system is controlled via two pathways innervating the liver ‐ one sympathetic and one parasympathetic, both originating from hypothalamic areas. Previous studies have shown that electrical stimulation of these hypothalamic nuclei can lead to rapid changes in blood glucose levels. Additional studies have also shown that electrical stimulation of peripheral nerves innervating the liver can lead to changes in hepatic output, however the limitations associated with neural control of glucose and lipid levels is unclear. Here we take an interdisciplinary approach to characterizing the use of electrical stimulation to control hepatic glucose and lipid metabolism. Using electrical stimulation, we demonstrate and characterize the ability to modulate both sympathetic and parasympathetic activity to the liver, enabling modulation of the glucose clearance rate assessed via oral glucose tolerances tests. Furthermore, we demonstrate how electrical stimulation can modulate lipid metabolism as a function of frequency, amplitude, and location of stimulation.Support or Funding InformationThis work is funded by the GlaxoSmithKline Bioelectronics Initiative.
Activation of dynamin‐related protein 1 contributes to conversion of cardiomyocytes into a proinflammatory phenotype in rat model of sepsis
Authors: Jiang Weipeng , Tao Aibin , Mele Tina , Kao Raymond , Martin Claudio , Rui Tao 
Published: November 2023

Dynamin‐related protein 1 (Drp1) is a regulator of mitochondrial fission and cell death pathways. The aim of this study is to determine whether Drp1 plays a role in conversion of cardiomyocytes into a proinflammatory phenotype in sepsis.MethodsIn vivo: rat model of sepsis was induced by intraperitoneally injection of fecal materials and myocardial MPO activity and PMN accumulation were assessed. In vitro: cultured rat cardiomyocytes were challenged with plasma (20%) of septic rats. Myocyte Drp1 activation (Western) and chemokine production (LIX and KC) were determined (RT‐PCR and ELISA).ResultsSix hours after induction of sepsis, PMN infiltration of the myocardial interstitium was evident in the septic rats. This inflammatory response was ameliorated by a Drp1 inhibitor, Mdivi‐1. Challenge of cultured rat cardiomyocytes with septic plasma resulted in Drp1 activation and converted the cardiomyocytes to a proinflammatory phenotype (the myocytes produced and secreted the chemokines, LIX and KC); effects blunted by Mdivi‐1.ConclusionThe results indicate that Drp1 plays an important role in conversion of cardiomyocytes into a proinflammatory phenotype. The Drp1 could be an effective target for treatment of sepsis. (*Corresponding author)Support or Funding InformationIRF of Lawson Health Research Institute (IRF 2014‐25); National Natural Science Foundation of China (81370333); Natural Science Foundation of Jiangsu Province, China (BK2015‐1332)
Breaking the TRPV1‐Calcineurin Interaction as a Novel Means for Pain Control
Authors: McAllister Stacy , Zambelli Vanessa , Gross Eric 
Published: November 2023

Targeting the transient receptor potential vanilloid 1 (TRPV1) channel for pain control is complex. Previous efforts in drug discovery resulted in failure since TRPV1 agonists are painful and TRPV1 antagonists cause hyperthermia. Calcineurin is an important TRPV1 protein partner since an interaction of TRPV1 with calcineurin favors TRPV1 channel opening. Therefore as an alternative, we hypothesized by breaking the interaction between TRPV1 and calcineurin, we can indirectly modulate TRPV1 to reduce pain sensation.To test this hypothesis, we used methods including computational biology, peptide synthesis, in vitro, and behavioral pain assays. Based on computational analysis, we discovered a sequence within the TRPV1 C‐terminus that interacts with the calcineurin A subunit. The TRPV1 region forms an alpha coil‐coil, part of the TRPV1 channel inner pore unit, and is evolutionary conserved in mammals. Based on these findings, we synthesized a peptide against this region, linked it to TAT47–57 and called it V1‐cal. In a competitive in vitro calcineurin activity assay, V1‐cal reduced the ability for calcineurin to dephosphorylate a calcineurin substrate as measured by free phosphate release in vitro. Further, we used a carrageenan‐induced inflammatory pain model in male Wistar rats of 10–14 weeks of age. Nociceptive threshold was assessed using the Randall‐Selitto method at baseline and 3 hours after administration of carrageenan. Ten minutes prior to carrageenan, rats were injected intraplantarly with the V1‐cal peptide or saline control. Relative to the control group, rats treated with the V1‐cal peptide had a higher mechanical pain threshold (55±8g vs 36±1g, mean±SEM, n=5/group). Together, these data suggest V1‐cal is an effective therapeutic for pain control by disrupting the TRPV1‐calcineurin interaction.Support or Funding InformationThis work was supported by NIH grant T32 GM089626 to SM.