bluuebunny/crossref_metadata_2025_split huggingface dataset:
ARH3 catalyzes degradation of mitochondrial matrixaccumulated Poly (ADP‐ribose)
Authors: Mashimo Masato , Niere Marc , Agledal Line , Dölle Christian , Kasamatsu Atsushi , Kato Jiro , Moss Joel , Ziegler Mathias 
Published: June 2021

Poly (ADP‐ribose) (PAR) metabolism, which is mainly mediated by members of the Poly (ADP‐ribose) polymerase (PARP) and Poly (ADP‐ribose) glycohydrolase (PARG) families, is involved in several cellular pathways including DNA stability, apoptosis, and mitosis. PARG, initially identified as a PAR degrading‐enzyme, exists as multiple isoforms with different subcellular localization, resulting from alternative splicing of the single gene. ADP‐ribosylhydrolase 3 (ARH3) also has PAR‐degrading activity. The enzymes participating in a putative PAR metabolism in mitochondria are still unknown.An alternatively spliced, small human PARG isoform (PARG55) and ARH3 are located in mitochondria owing to a mitochondrial targeting sequence at their N‐terminus. Using a modified PARP1 construct to produce PAR selectively in mitochondria, ARH3‐deficient cells were shown to have more mitochondrial PAR accumulation than wild‐type cells. Overexpression of ARH3 decreased mitochondrial PAR content, while overexpression of PARG55 had no effect. PARG55 was shown to lack exon5, which is essential for PAR‐degrading activity. These data suggest that ARH3 rather than PARG55 is the primary enzyme involved in catalyzing the degradation of PAR in mitochondria.
LPS/TLR4‐NF‐[kappa]B axis signaling amplifies STIM1 expression to augment PAR‐1‐induced Calcium entry and permeability response in lung microvessels
Authors: DEBROY AUDITI , Sundivakkam Premanand , Singh Vandana , Malik Asrar B , Tiruppathi Chinnaswamy 
Published: June 2021

RationaleHuman sepsis is characterized by lung microvascular leakage. Store‐operated Calcium Entry (SOCE) in endothelial cells (ECs) mediates increase in lung vascular permeability. Here we investigated the role of the transcription factor NF‐κB, activated downstream of TLR4 in mediating the expression of the crucial activator of SOCE, Stromal Interacting Molecule 1 (STIM1), and its consequences on PAR‐1‐mediated Ca2+ entry in ECs and lung vascular leak.ResultsLPS induced STIM1 expression in lung ECs. Increased STIM1 expression was associated with augmented PAR‐1‐mediated SOCE and permeability increase. These results were recapitulated in vivo mouse model LPS‐induced sepsis and the response was abrogated in TLR4 knockout mice. To characterize the transcriptional mechanism of LPS induction of STIM1, we analyzed the 5′‐regulatory region of STIM1 gene and found the consensus binding sites for LPS‐sensitive transcription factors NF‐κB, SP1, and AP1. By electrophoretic mobility shift assay, we observed the binding of NF‐κB and SP1 to the hSTIM1 promoter. We cloned the hSTIM1 promoter and showed LPS induction of promoter activity in ECs. Silencing of p65/RelA markedly reduced LPS‐induced STIM1 expression and subsequent PAR‐1 responses in ECs.ConclusionThese results collectively suggest the novel and critical role of STIM1 in mediating lung vascular hyper‐permeability during gram‐negative sepsis.
Rapid and Specific Measurements of Superoxide Using Fluorescence Spectroscopy
Authors: Nazarewicz Rafal , Bikineyeva Alfiya , Harrison David G , Dikalov Sergey 
Published: June 2021

Superoxide plays a critical role in number of pathological conditions including cardiovascular diseases. Development of specific and reliable methods for superoxide detection is critical for investigation of these conditions. Cellular reactions of dihydroethydium (DHE) produce both the superoxide specific product 2‐OH‐ethidium (2‐OH‐E) and non‐specific oxidation product ethidium which have overlapping fluorescence spectra. Specific superoxide detection therefore is difficult. HPLC can separate ethidium and 2‐OH‐E but this method is limited by inability to detect site‐specific 2‐OH‐E and duration of the HPLC procedure. In this study, we show optimized fluorescence spectroscopy protocol allowing reliable superoxidedetection in cell free systems, subcellular fractions, intact cells and tissue. Analysis of fluorescence emission spectra showed four times higher contribution of 2‐OH‐E fluorescence comparing to ethidium when excitation was changed from 480 nm to 405 nm. Selection of specific emission enables selective detection of 2‐OH‐E with negligible contribution of ethidium. The best selectivity of fluorescence signal for detection of 2‐OH‐E at excitation at 405 and 480 nm was achieved at emission 570 nm (1:21 ethidium:2‐OH‐E ratio) and 530 nm (1:52 Ethidium:2‐OH‐E ratio), respectively. We applied these optimized conditions for on‐line superoxidedetection generated by xanthine oxidase. Fluorescent signal was inhibited by SOD supplementation confirming specific detection of 2‐OH‐E with xanthine oxidase concentration as low as 10 microunits per ml. Specificity of plate reader‐based superoxide measurements was confirmed by HPLC analysis of 2‐OH‐E and ethidium. Our study shows that limitations of HPLC in superoxide detection in biological samples can be substantially overcome by using optimized fluorescence spectroscopy.This work has been funded by National Institute of health PO‐1 HL058000.
Identification of the Differentiation Status of Individual Hematopoietic Cells from Mouse Bone Marrow using Secondary Ion Mass Spectrometry
Authors: Kraft Mary L , Frisz Jessica F. , Choi Ji Sun , Wilson Robert L. , Harley Brendan A. 
Published: June 2021

The ability to self‐renew and differentiate into multiple types of blood and immune cells renders hematopoietic stem cells (HSCs) of significant value for tissue engineering applications. To enable engineering stem cell fate within laboratory settings, the combination of chemotactic and micromechanical properties that direct HSC maturation must be elucidated. This task requires determining the differentiation stage of individual cells within ex vivo culture systems. New experimental approaches are required to identify the differentiation stage of individual cells and to correlate this information on maturation stage with cell location within their environment. Here we demonstrate that multivariate analysis of time‐of‐flight secondary ion mass spectrometry (TOF‐SIMS) data can be used to successfully classify individual hematopoietic cells from mouse bone marrow according to their distinct lymphopoiesis phenotype, specifically HSCs, common lymphoid progenitors, or mature B cells.This research was supported by a Career Award at the Scientific Interface from the Burroughs Wellcome Fund.
Selenoprotein W is required for epidermal growth factor (EGF)‐stimulated proliferation via a pathway involving MKK4 and p53
Authors: Hawkes Wayne Chris , Alkan Zeynep 
Published: June 2021

Selenoprotein W (SEPW1) is a highly conserved thioredoxin‐like protein whose depletion causes a p53‐ and p21‐dependent G1‐phase cell cycle arrest in breast and prostate epithelial cells. SEPW1 promotes the ubiquitination, nuclear export, and proteasomal degradation of p53, effects that are associated with decreased phosphorylation of Ser‐33 in p53. We now report that SEPW1 is required for EGF‐stimulated proliferation in breast and prostate cells. SEPW1 depletion blocked EGF‐stimulated G1 to S‐phase progression and altered signaling via downstream MAP kinase pathways. In an siRNA screen of 4 MAP kinase kinases and 6 MAP kinase kinase kinases, only silencing of MKK4 (JNKK1/SEK1) blocked G1 arrest, Ser‐33 phosphorylation, and p53 activation from SEPW1 depletion in EGF‐stimulated cells, suggesting MKK4 is downstream from SEPW1 in a p53‐dependent pathway regulating EGF‐stimulated cell cycle entry. We found that silencing p53 in freely cycling cells accelerated G1 to S‐phase progression, suggesting SEPW1 stimulates proliferation by suppressing the already low basal level of p53 expression. Taken together, these results suggest SEPW1 mediates EGF‐stimulated proliferation, at least in part, by suppression of p53 via MKK4, and presumably, one or more downstream MAP kinases.This research was supported by USDA CRIS Project 5306‐51530‐018‐00D. USDA is an equal opportunity provider and employer.
Regulation of macrophage cholesterol homeostasis by trans fatty acid
Authors: Shao Fei , Ford David A. 
Published: June 2021

Trans fatty acid (TFA) consumption causes adverse changes in plasma lipids and lipoprotein profile, which leads to increased incidence of atherosclerosis. Macrophage cholesterol efflux, an atheroprotective process, is mediated by ATP binding cassette transporter A1 (ABCA1). ABCA1 is highly regulated by transcription factors Liver X receptor‐α (LXRα) and Retinoic X Receptor‐α (RXRα). Our studies are focusing on the mechanism by which individual trans fatty acid molecular species regulate ABCA1‐mediated macrophage cholesterol efflux and macrophage cholesterol efflux. Our results show that trans fatty acid species differentially regulate ABCA1 protein expression in mouse macrophage J774. The most abundant trans fatty acid elaidic acid (9t‐18:1) protects ABCA1 protein degradation, whereas oleic acid (9c‐18:1) induce ABCA1 protein degradation. Elaidic acid upregulates LXRα/RXRα activity and sterol regulatory element (SRE)‐driven luciferase activity, which suggest that elaidic acid may increase activities of sterol regulatory element binding proteins (SREBPs) family. These results demonstrated that trans fatty acids regulate ABCA1‐mediated cholesterol efflux and cholesterol homeostasis in a distinctive way compared to cis fatty acids.This research was supported by NIH grants HL074214 and HL098907 (DAF)
Functional and structural analysis of transmembrane segment IV of Na+/H+ exchanger of Schizosaccharomyces pombe
Authors: Ullah Asad , Alves Claudia , Lee Brian L , Sykes Brian D , Fliegel Larry 
Published: June 2021

Sod2 is the plasma membrane Na+/H+ exchanger of fission yeast S. pombe. It plays an important role in providing salt tolerance to S. pombe by removing excess intracellular sodium (or lithium) in exchange for protons. We examined the role of amino acids residues of TMIV (126FQINFLGSLLIAGCITSTDPVLSALI152) in activity of this protein using alanine scanning mutagenesis, and examining LiCl or NaCl tolerance in Sod2‐deficient S. pombe. We identified two amino acids, T144 and V147, critical for the function of Sod2. The mutations T144A and V147A resulted in a defective sod2 protein that did not confer salt tolerance when reintroduced into to S. pombe. Sod2 protein with other mutations in TMIV, exported either sodium or lithium as efficiently as the wild type Sod2 or had intermediate effects on salt tolerance of S. pombe. When we mutated T144 to aspartic acid, lysine or serine, the mutant proteins were impaired in their ability to allow growth in both lithium and sodium containing media, comparable to the knock out sod2 yeast (sod2::ura4). Western blot analysis showed that mutants T144A and V147A were expressed at comparable levels as the wild type sod2. To determine the structure of TMIV, we expressed and purified TMIV as a fusion protein with maltose binding protein. The TMIV peptide was released by cleaving the fusion protein with tobacco etch virus and re‐purified by HPLC. The structure is being determined by NMR.Supported by Natural Sciences and Engineering Research Council of Canada
The Vibrio parahaemolyticus effector VopQ induces autophagosome accumulation in host cells
Authors: Sreelatha Anju , Bennett Terry , Starai Vincent , Orth Kim 
Published: June 2021

Vibrio parahaemolyticus, a Gram‐negative bacterium, is the leading cause of seafood‐borne illness in the United States. This pathogen is able to infect and survive in humans by manipulating the host cellular machinery via bacterial Type III effector proteins that are translocated into the host cell, where they disrupt cellular signaling. One of the principal effector proteins, VopQ, has been demonstrated to be necessary and sufficient to induce autophagosome accumulation. I propose to elucidate the precise cellular and molecular mechanism by which VopQ induces autophagosome accumulation. Since autophagosomes are transient vesicles, the accumulation could be due to an increase in formation of autophagosomes or a decrease in lysosomal fusion. Using biochemical assays and yeast genetics we have uncovered a novel target that reveals the role of VopQ in vesicle fusion during autophagy.With the discovery of several bacterial effectors that function to manipulate host membranes, many eukaryotic vesicle trafficking pathways have been unveiled, and many new questions have surfaced about the mechanisms of novel bacterial effectors. Understanding the mechanism by which VopQ induces autophagosome accumulation could be essential in understanding V. para infection and could give new insights into the mechanisms of eukaryotic vesicle trafficking.
Antibacterial Activity of Some Coumarins and their Metal Complexes
Authors: Govori Sevdije R , Buza Aida , Ajvazi Njomza , Haziri Arben 
Published: June 2021

A series of some new coumarin derivatives and their Cu(II) complexes have been synthesized and characterized by a variety of physicochemical techniques. The ligands and its metal complexes are screened for their in vitro antibacterial activity against Escherichia coli and Enterochoaco.The studies demonstrated that metallation can increase the antibacterial activity rather than the free ligand.
Effect of dairy foods on cell adhesion molecules (CAMs)
Authors: Hilpert Kirsten , West Sheila G , Ferencik Emily , Bagshaw Debrah , Kris-Etherton Penny M 
Published: June 2021

CAMs mediate atherosclerosis and thus are considered biomarkers of endothelial dysfunction. Elevated levels of CAMs predict heart disease risk and research has focused on strategies to lower them. Yet, studies of how diet affects CAMs are lacking. This study examined the effect of dairy foods on CAMs serum levels. Adults with untreated hypertension (n=23) were fed 3 experimental diets (5 wk each) in a randomized crossover study design. Diets included a dairy‐rich, high fruits and vegetables diet (D‐F&V; 30% fat, 7% saturated fat [SFA], 3.4 servings [svg]/d dairy, 9.6 svg/d F&V), a high fruits and vegetables diet (F&V; 30% fat, 7% SFA, 0.4 svg/d dairy, 9.6 svg/d F&V), and an average Western diet (AWD; 36% fat, 15% SFA, 0.4 svg/d dairy, 3.6 svg/d F&V). The D‐F&V diet reduced VCAM‐1 (547 ± 33 ng/mL) by about 16% compared with the F&V diet (649 ± 33 ng/mL, p < 0.05) and the AWD diet (650 ± 34 ng/mL, p < 0.05). Compared with the AWD diet, ICAM‐1 (242 ± 14 ng/mL) was significantly reduced by both the D‐F&V (225 ± 14 ng/mL, p < 0.05) and F&V diets (226 ± 14 ng/mL, p < 0.05). E‐selectin and P‐selectin followed a similar pattern, with the D‐F&V diet eliciting the lowest levels (D‐F&V vs. AWD, p < 0.02; D‐F&V vs. F&V, p < 0.1; F&V vs. AWD, p=ns). Collectively, these results suggest that dairy foods may have beneficial effects on endothelial function in hypertension.Funding: National Dairy Council, GCRC Penn State University NIH M01 RR10732
Sacha inchi (Plukenetia volubillis L.) oil as novel source of dietary fat to prevent inflammation and cardiovascular disease risk
Authors: Noratto Giuliana Doris , Chirinos Rosana , Campos David , Larondelle Yvan , Arbizu Shirley , Dos Santos Manoela Maciel , Mertens-Talcott Susanne 
Published: June 2021

Cardiovascular‐related diseases (CVD) are the main cause of death around the world and the largest economic burden. Western diets have adopted a pattern in which ratios omega‐6 (n‐6): omega‐3 (n‐ 3) fatty acids (FA) are 15:1–16:1, which promotes inflammation and CVD. Sacha inchi seeds, native to Peru and source of dietary oil (SIO) contain a FA profile of n‐3: n‐6 ≈ 1, characteristic in Mediterranean diet. However, the composition of SIO varies by cultivar and the health benefits have not been investigated. Our goal was to characterize SIO in 16 cultivars and the antiinflammatory properties using human endothelial vein cells (HUVEC). SIO were analyzed by gas chromatography and HPLC, and antioxidant activity (AOX) by ABTS. A commercial SIO was assessed in HUVEC using the dichlorofluorescein diacetate (DCFH‐DA) assay for reactive oxygen species (ROS), and qRT‐PCR for mRNA. Results showed that SIO from different cultivars are rich in (n‐6) linoleic (36.1–40.9%) and (n‐3) α‐linolenic acids (37.2–43.9%), γ‐(702.6–1601.3 μg/g), and δ‐tocopherols (0.8 y 925.9 μg/g) and exerted AOX (7.7–10.3 μmol TE/g). SIO decreased H2O2‐induced ROS (0.73‐fold) and downregulated LPS‐induced expression of NF‐kB (0.84‐fold), TNF‐α (0.72‐fold), VCAM‐1(0.68‐fold), and ICAM‐1 (0.64‐fold). Overall, these results suggest the health cardioprotective benefits of SIO as a novel source of dietary fat.Grant Funding Source : ASN
Glucosamine regulates autophagy in vitro and in vivo
Authors: Carames Beatriz , Lotz Martin 
Published: June 2021

ObjectiveTo determine whether glucosamine (GlcN), a dietary supplement that is safe and beneficial in cellular and animal models of joint health, can activate autophagy, an essential cellular homeostasis mechanism that was found deficient in ageing and osteoarthritis (OA) cartilage.MethodsChondrocytes from normal human joints were treated with GlcN‐HCl (1 or 10 mM). Autophagy activation and mTOR signaling was determined by Western Blotting. Reporter mice with green fluorescent protein fused to the autophagy marker LC3 (GFP‐LC3 mice) were treated with GlcN‐HCl (250 mg/kg weight/day) for 7 days. Autophagy activation was analyzed by confocal microscopy and immunohistochemistry.ResultsGlcN treatment of chondrocytes induced autophagy by increasing LC3‐II expression, a main marker of autophagy. This was accompanied by inhibition of ribosomal protein S6 phosphorylation (prbS6), a target of mTOR. In GFP‐LC3 mice, gavage administration of GlcN led to a marked activation of autophagy in liver. In articular cartilage and menisci, there was a decrease in prbS6 expression.ConclusionsGlcN modulates several components of the autophagy pathway in vitro and in vivo. These findings suggest that GlcN is an effective autophagy activator and motivate future studies on its efficacy in modifying aging‐related cellular changes and supporting joint health.Support: Funding provided by Cargill, Inc.
Effects of Vitamin D and Calcium Supplementation on Blood Pressure and Serum Lipids: A Randomized, Double‐blind, Placebo‐controlled, Clinical Trial
Authors: Chai Weiwen , Bostick Roberd M , Franke Adrian A , Cooney Robert V 
Published: June 2021

ObjectiveTo estimate the effects of vitamin D +/− calcium supplementation on blood pressure and serum lipid levels.MethodsNinety‐two men and women (40 to 75 years) were randomized in a pilot, double‐blind, placebo‐controlled clinical trial of daily supplemental vitamin D3 800 IU and calcium 2 g (as calcium carbonate) alone or in combination in divided doses twice daily with meals. Blood was drawn at baseline and after 6 months of intervention and stored serum samples were analyzed for serum total cholesterol and triglycerides. Systolic and diastolic blood pressures were also measured at baseline and at one and six months follow up.ResultsRelative to placebo, mean serum triglycerides decreased 30.1% (P = 0.20) and 32.1% (P = 0.18) in the calcium and calcium plus vitamin D3 treated groups, respectively. Similar but non‐substantial decreases were observed for mean serum total cholesterol levels (calcium group, 5.3% lower, P = 0.31; calcium plus vitamin D3 group, 6.8% lower, P = 0.19). In the intention‐to‐treat analyses there were no statistically significant changes in mean systolic and diastolic blood pressures in the calcium and vitamin D groups relative to the placebo group.ConclusionsSupplemental vitamin D3 and calcium may reduce serum lipid levels but have no substantial effects on blood pressure.
Exploring the association between dietary patterns and the presence of Age‐related Macular Degeneration
Authors: Goulão Beatriz , Camacho Pedro Miguel , Mota Carla , Coelho Inês , Roe Mark , Castanheira Isabel 
Published: June 2021

Age‐related Macular Degeneration (AMD) is the leading cause of irreversible blindness in Europe. The role of nutritional factors in the prevention of disease are not clear, in part due to difficulties in obtaining comparable nutrient intake values, in international, multi‐centre studies. EuroFIR (European Food Information Resource) eSearch platform, including more than twenty national food composition databanks, was used to examine whether specific nutrient antioxidants, fat, protein, and carbohydrates from natural food sources of different geographic origins accounted for association between AMD and dietary patterns. A cohort study was used. Participants, caucasian men and women aged over 55 selected by a AMD specialist, completed the FFQ in an interview mode during their clinical visit to a specialized eye center.In patients who consumed a diet rich in total fat, a correlation between the choroid thickness and AMD was found (p‐value< 0,001). Clinical Nutrition studies, covering natives from Sao Tome and Principe are in progress as a contribution to understand if racial differences associated with dietary patterns affected the pathogenesis of AMD and other eye diseases.
Development of a Four‐Week Cyclic Mexican Mediterranean Menu for Health
Authors: Duran Carla Dee , Tam Chick F. 
Published: June 2021

The mixed genetics of many Mexican Americans and Mexicans have compromised folate metabolism due to possible C677T mutation. These mutants that originate from the Mediterranean regions need to consume Mediterranean region diets to survive. However, when these mutants immigrate to the New World and eat the Western diet, they begin to acquire Metabolic Syndrome (MetS). The objective is to develop a four‐week cyclic menu that contains high contents of plant foods, fish, and olive oil (the Mediterranean diet) combined with the foods established by the indigenous people of the Americas with high antioxidants and fibers to reduce the risk of MetS. First, the nutrients of each day were analyzed by diet computer program. Second, an average of 28‐days nutrient analysis was obtained to demonstrate nutrient adequacy. Third, the functional ingredients and their biochemical/biological active molecules were obtained: quinoa (saponins like solanine for heart health), amaranth (stanols and squalene to lower cholesterol), cilantro (4‐hydroxycoumarin, a vitamin K antagonist), anti‐inflammatory agents found in avocado, pumpkins, yams (carotenoids, lutein/zeaxanthin, flavonoids, anthocyanins, and proanthocyanidins), and nopales cactus: Opuntia (candicine, hordenine, betalains). The anti‐inflammatory properties of the diet can have the protective effects against the MetS found in Mexican Americans and Mexicans.No source of financial research support.