bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cargo Receptor Traffics Many Matrix Proteins Out of the Endoplasmic Reticulum (ER) by Binding to N‐Terminal IleProVal (IPV)‐Like Motif
Authors: Yin Ying , Garcia Mekka , Novak Alex , Fisher Larry 
Published: June 2021

IntroductionMatrix proteins assemble large complexes in the extracellular environment and may be at risk of forming undesired aggregates during intracellular trafficking. Patients with nonsyndromic Dentinogenesis Imperfecta have mutations in the dentin sialophosphoprotein (DSPP) gene. In half of the cases, the N‐terminal IPV tripeptide is changed (e.g., IPD, ITV). We have shown that IPV‐mutant DSPP inefficiently exits the ER and forms aggregates. Many secreted proteins of dentin (e.g., DSPP, DMP1), enamel (AMELX, AMBN, ENAM), bone (SPARC, OCN, OPN) and cartilage (BGN, DCN, ASPN), as well as enzymes (trypsin) and hormones (GH, LEP, PRL, FGF23) also start with conserved IPV‐like tripeptides. We propose that the same cargo receptor binds to these proteins through their IPV‐like motifs thereby trafficking them out of the ER before self‐ or inter‐molecular aggregates form.MethodsThe ER‐trafficking efficiency of DSPP (and other secreted proteins) expressing a series of single amino acid changes in their IPV motifs was studied in HEK293 and HeLa cells. To identify the cargo receptor, we expressed wildtype and mutant constructs in Saccharomyces cell lines each lacking a single ER‐associated protein.ResultsWe have defined the position/property of amino acids in IPV‐like motif that promote ER exit in yeast and higher eukaryotes. Furthermore, gene deletion of a conserved ER‐associated yeast protein specifically delays ER exit of wildtype yeast and transfected matrix proteins.ConclusionWe have defined the amino acids required to bind the proposed cargo receptor and have identified the yeast's receptor as a highly conserved, ER‐associated protein.Supported by Division of Intramural Research of the NIDCR, NIH.
Extracellular Matrix Fibronectin Attenuates PDGF‐Induced Intracellular Calcium Release
Authors: Farrar Christopher , Hocking Denise 
Published: June 2021

The extracellular matrix (ECM) has structural and signaling roles within tissues. Fibronectin is an ECM protein that is secreted by fibroblasts in a protomeric form and assembled into a fibrillar matrix by a cell‐mediated process termed fibronectin matrix assembly. ECM proteins affect growth factor signaling. Platelet derived growth factor (PDGF) is a major mitogen for fibroblasts that promotes proliferation and migration. The objective of this study was to determine the effect of ECM fibronectin on the response of fibroblasts to PDGF. The response of cells to PDGF was quantified by measuring intracellular calcium release in Fluo‐4 loaded cells. Calcium release was measured in fibronectin‐null mouse embryonic fibroblasts, allowing for control over the amount of fibronectin to which cells were exposed and the initiation of fibronectin matrix assembly. Results showed that in the absence of fibronectin, addition of 50 ng/mL PDGF to cells transiently increased intracellular calcium concentration. In contrast, incubation of cells with 25 nM fibronectin for 20 hr significantly attenuated the PDGF‐induced increase in calcium release. Fibronectin‐mediated attenuation of PDGF‐induced calcium release was blocked by an inhibitor of fibronectin matrix assembly, indicating that the ECM form of fibronectin is required for fibronectin‐mediated attenuation of PDGF signaling. A fibronectin fragment engineered to mimic the ECM form of fibronectin also attenuated PDGF‐induced calcium release. Attenuation of PDGF‐induced calcium release was not due to downregulation of PDGF receptor β expression. This study demonstrates that fibronectin matrix assembly attenuates PDGF‐induced intracellular calcium release by an as‐yet unidentified mechanism.Supported by: NIH/NHLBI: T32 HL066988
MAP1S‐enhanced Autophagy Alleviates Tissue Fibrosis and Sustains Mouse Survival under Stress Imposed by LC3‐induced Overexpression of Fibronectin
Authors: Liu Leyuan 
Published: June 2021

Autophagy is a cellular process that controls and executes the turnover of misfolded and abnormally aggregated proteins. Microtubule‐associated protein 1S (MAP1S) interacts with autophagy marker light chain 3 (LC3) and positively regulates autophagy flux. LC3 also binds with fibronectin mRNA and facilitates its sorting along microtubules onto rough endoplasmic reticulum where it is translated into fibronectin protein. Fibronectin is exported to surface of plasma membrane to initiate the assembly of fibronectin extracellular matrix and other extracellular matrix proteins. Fibronectin matrix is degraded in lysosomes after it is engulfed into cytosol via endocytosis. Here we show that overexpression of LC3 in wildtype mice promoted the synthesis and degradation of fibronectin and consequently resulted in a balanced levels of fibronectin; therefore, the GFP‐LC3 transgenic mice had a normal length of lifespan as the wild‐type mice did. In GFP‐LC3 transgenic mice with MAP1S deleted, LC3 overexpression enhanced synthesis of fibronectin while MAP1S‐deficiency‐caused autophagy defect impaired the degradation of fibronectin. The accumulation of fibronectin protein led to enhanced levels of oxidative stress, genomic instability and tissue fibrosis, and to a dramatic 35% reduction in mouse lifespan. Therefore, autophagy may play an important role in maintaining mouse lifespan via regulating the homeostasis of fibronectin extracellular matrix.This work was supported by a NIH R01CA142862 to Leyuan Liu.
Evaluating p97 inhibitor analogues for their domain‐selectivity and potency against the p97‐p47 complex
Authors: Gui Lin , Zhang Xiaoyi , Fang Chenjie , Moen Derek R. , Wang Daniel , Chou Tsui‐Fen 
Published: June 2021

The p97 AAA is an important therapeutic target for cancer and neurodegenerative diseases. Each monomer is composed of an N domain followed by D1 and D2 ATPase domains. It is clear that the D2 domain hydrolyzes ATP in vitro. D1 shows ATPase activity in the full‐length protein only when the D2 domain is capable of binding nucleotides. A major role of the N domain is to recruit numerous p97 cofactors. Binding of p47‐one of the major p97 cofactors‐ to the N‐domain of p97 significantly inhibits wild type p97 ATPase activity.Based on observations of enzymatic activity with small‐molecule inhibitors, we have recently evaluated that four p97 inhibitors have differential responses to disease‐causing IBMPFD mutations and to the presence of the N domain binding cofactor protein p47. DBeQ and NMS‐873 inhibited both ATPase domains, whereas ML240 and ML241 specifically target the D2‐domain of wild type p97. In addition, p47 significantly decreases their potency by ~50‐fold. In contrast,DBeQshows only a 4‐ to 6‐fold decrease in potency against the p97‐p47 complex.To elucidate the structure‐activity relationships of 200 potential p97 inhibitor analogues, we characterize their ability to inhibit the p97 ATPase activity of the D1, D2, or both domains as well as the effect of p47 on potency. The selectivity of 29 of these compounds was further examined by 8‐dose titration.These studies identify several potential starting points for future SAR analyses and also highlight the possibility of developing domain‐selective and complex‐specific p97 inhibitors. Together, our results provide the framework for developing domain, complex, and pathway specific inhibitors, with the ultimate goal of validating p97 as a potential therapeutic target.
Synthesis and Characterization of Triazine‐based Chemical Probes
Authors: Cole Kyle , Banerjee Ranjan , Pace Nicholas , Weerapana Eranthie 
Published: June 2021

Rapid proliferation of cancer cells requires upregulated protein synthesis. Protein folding in the cell is assisted by chaperones and foldases– that accelerate protein folding and inhibit aggregation. One such family of chaperones is the protein disulfide isomerase (PDI) family. The first PDI discovered, PDIA1, is a 57‐kDa protein which is localized in the ER and has been shown to be critical for tumor growth in a mouse xenograft model of ovarian cancer. Several small molecule inhibitors of PDIA1 have been developed in the last few years, including one from our lab, RB‐11‐ca; which is equipotent, yet more site‐selective than the best current commercially available inhibitor, 16F16. Current efforts are to synthesize novel analogs of RB‐11‐ca to increase potency and selectivity against PDIA1. One compound, KSC‐7, has shown approximately a 20% increase in affinity for PDIA1 compared to RB‐11‐ca.The triazine‐based scaffold has also become useful for targeting reactive lysines. A dichlorotriazine electrophile, RB7, has been discovered to label lysines, however, it is far too reactive for covalent‐inhibitor design. Efforts to replace the second chlorine with an amine substitution, to tune down the reactivity of the electrophile, led to a complete loss of protein labeling, Therefore, if we substitute a less electron donating substituent in the second position, then we would be able to tune down the reactivity of the electrophile enough to maintain the reactivity, but be able to selectively target reactive lysine residues.
Curcumin Inhibits DLD‐1 Colorectal Cancer Cell Proliferation by Modulating MicroRNA and Tumor Suppressor Gene Expression
Authors: Adeyeni Temitope , Vattathara Kenn , Duggirala Rajat , Selvamani Vijayalingam , Ezekiel Uthayashanker 
Published: June 2021

Phytochemicals are plant‐based, non‐toxic compounds that produce health benefits. Curcumin, a major phytochemical in turmeric, inhibits the proliferation of many types of cancer cells, both in vitro and in vivo. Phytochemicals subvert carcinogenesis by modifying methylation patterns normally maintained by cancer cells. This methylation modification mechanism indirectly influences the regulatory microRNA that maintains normal cell division. Earlier, we identified that curcumin exerts an antiproliferative effect on DLD‐1 colon cancer cells in a dose dependent manner. After 48‐hour treatment of DLD‐1 cells with curcumin, expression level of tumor suppressor gene, RASSF1A, and microRNA‐15a (miR‐15a) was evaluated. Normally, expression levels of both RASSF1A and miR‐15a were suppressed in DLD‐1 cells; however, upon curcumin treatment, expression levels of RASSF1A and miR‐15a were higher than those in vehicle‐treated controls. Also, the miR‐15a target, BMI1, was suppressed in curcumin‐treated cells. In summary, curcumin exerted an antiproliferative effect against colon cancer cells, possibly by an epigenetic mechanism.KV and RD were supported by the DeNardo Foundation Fellowship.
­Role of Cytochrome c Phosphorylation in Regulation of Respiration and Apoptosis
Authors: Mahapatra Gargi , Varughese Ashwathy , Qinqin Ji , Lee Icksoo , Salomon Arthur , Huttemann Maik 
Published: June 2021

Cytochrome c is a functionally diverse protein, carrying electrons in the electron transport chain and playing a critical role in cellular apoptosis. Previous work by our lab suggest the importance of reversible phosphorylation of cytochrome c in regulating its functions. We hypothesize that under healthy conditions, phosphorylated cytochrome c partially inhibits mitochondrial respiration and maintains healthy mitochondrial membrane potential, preventing ROS generation, while cellular stress‐mediated dephosphorylation leads to increased respiration and ROS generation, initiating apoptosis.In this study, we purified bovine kidney cytochrome c in the presence of phosphatase inhibitors, identified threonine phosphorylation by immunoblot analysis, and determined threonine 28 phosphorylation by Nano/LC/ESI/MS/MS. To characterize its effect, Thr28 was mutated to glutamate, a phosphomimetic mutation, and alanine, a nonphosphorylatable control, and experiments were performed with bacterially overexpressed proteins. In the reaction with cytochrome c oxidase, Thr28Glu showed significant reduced respiration rates compared to wild‐type, with no alteration in caspase‐3 activation. Thr28Glu had a higher reduction rate from an initial oxidized state, with no change in the rate of oxidation compared to wild‐type, and was found to be resistant to heme destruction in the presence of excess ROS. Our data suggest that Thr28, conserved in mammalian cytochrome c, is an important regulatory site.
MUC1‐C Induces the LIN28B→LET‐7→HMGA2 Axis to Regulate Self‐renewal in NSCLC
Authors: Ahmad Rehan , Alam Maroof , Rajabi Hasan , Kufe Donald 
Published: June 2021

ObjectiveThe LIN28B→let‐7 pathway contributes to regulation of the epithelial‐mesenchymal transition (EMT) and stem cell self‐renewal. The oncogenic MUC1‐C transmembrane protein is aberrantly overexpressed in lung and other human carcinomas; however, there is no known association between MUC1‐C and the LIN28B→let‐7 pathway. Methods: Stable silencing of MUC1‐C using MUC1 shRNA. Targeting MUC1‐C function with a dominant‐negative mutant or a peptide inhibitor was used to study the link between MUC1 expression and LIN28B→let‐7 signaling. Results: In non‐small cell lung cancer (NSCLC), silencing MUC1‐C downregulates the RNA binding protein LIN28B and coordinately increases the miRNA let‐7. Targeting MUC1‐C function with a dominant‐negative mutant or a peptide inhibitor provides confirming evidence that MUC1‐C induces LIN28B→let‐7 signaling. MUC1‐C promotes NF‐κB p65 chromatin occupancy of the LIN28B first intron and activates LIN28Btranscription which is associated with suppression of let‐7. Consistent with let‐7‐mediated inhibition of HMGA2 transcripts, targeting of MUC1‐C also decreases HMGA2 expression. HMGA2 has been linked to stemness, and functions as a competing endogenous RNA (ceRNA) of let‐7‐mediated regulation of the TGFβ co‐receptor TGFBR3. Accordingly, targeting MUC1‐C suppresses HMGA2 mRNA and protein which is associated with decreases in TGFBR3, reversal of the EMT phenotype and inhibition of self‐renewal capacity. ConclusionThese findings support a model in which MUC1‐C activates the ⇑LIN28B→⇓let‐7→⇑HMGA2 axis in NSCLC and thereby promotes EMT traits and stemness.
Epigenetic Regulation of RGS2 in Cystic Fibrosis
Authors: Bouvet Guillaume , Cyr Yannick , Massé Chantal , Berthiaume Yves 
Published: June 2021

Cystic fibrosis (CF) is a recessive autosomal disease that affects the respiratory, gastrointestinal and reproductive systems. At pulmonary level, the absence of CFTR function (cystic fibrosis transmembrane conductance regulator encoding an ion channel chlorine) leads to a decrease in mucociliary clearance, resulting in mucus accumulation. A new approach based on epigenetic regulation of gene expression has been investigated and we identified a list of hypermethylated genes in dF508 cell lines, the most frequently observed mutation in the CF population. Among the CF hypermethylated genes, RGS2 (Regulator of G protein signaling 2) gene demonstrated either transcript or protein downregulation. RGS2 is a key modulator of bronchial hyperresponsiveness, plays a critical role in fibrosis‐related diseases and inflammation.We hypothesized that variation in RGS2 protein expression could be the result of modulation of promoter methylation and has an impact on cytokine regulation. The first objective was to verify that methylation observed on RGS2 promoter in dF508 homozygous cell lines induces a modulation of protein expression. Then, to assess the consequences of this downregulation of RGS2, we performed an shRNA‐mediated RGS2 knockdown followed by DMNQ treatment in non‐CF cells and monitored the expression of inflammatory mediators.We confirmed that methylation of RGS2 promoter has an impact on RGS2 gene expression using azacythidine and oxidative stress treatments and that RGS2 is downregulated at the protein level in CF cells and in dF508 homozygous patients. We observed a relation between RGS2 downregulation by shRNA and the upregulation of 3 modulators of early inflammation: CHI3L1, S100A12 and TNFa.
Production of transgenic mice expressing EGFP under monkey 20α‐HSD promoter
Authors: Min Kwan‐Sik , Park Chae‐Won , Seo Eun‐Bi , Kang Myung‐Hwa 
Published: June 2021

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy, if it occurs. Progesterone primes the endometrium for possible implantation and inhibits uterine contraction until birth. Aldo‐keto reductases (AKRs) belong to a superfamily of NADPH‐dependent reductases that act on a wide range of substrates, including simple carbohydrates, steroid hormones, and endogenous prostaglandins. 20‐alpha hydroxysteroid dehydrogenase (20α‐HSD; EC.1.1.1.149) enzyme belongs to the family of aldo‐keto reductases. 20α‐HSD predominantly converts progesterone into its biologically inactive form 20α‐hydroxyprogesterone (20α‐OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. We have been reporting on the molecular characterizations of placental and ovarian 20α‐HSD in the bovine, pig, deer and monkey.In this study, to understand the fundamental function of monkey 20α‐HSD, we created transgenic mice expressing EGFP gene under monkey 20α‐HSD promoter. The EGFP protein was expressed in the placenta and ovary of tg mice during pregnancy. Now, we are trying to elucidate the function by PGF2α, Prolactin, Oxytocin. (Eun‐Bi Seo and Chae‐Won Park were supported by a scholarship from the BK21 Plus Program (31Z20130012928). the Ministry of Education, Science and Technology, korea)
Short Pictorial Sweetened Beverage Scale Demonstrates Convergent Validity with Diet Logs
Authors: Townsend Marilyn , Shilts Mical , Styne Dennis , Aden Christopher , Drake Christiana 
Published: June 2021

Many nutrition education programs target soda in an effort to improve behaviors shown to be related to diet quality and weight status. Using visually enhanced behavioral items designed for low literate participants of USDA food assistance and education programs, our objective is to provide evidence of convergent validity with dietary recall logs. Low‐income parents (n=144) provided self‐administered Healthy Kids (HK) and three child 24‐hour diet recalls. A sweetened beverage variable was the sum of the ounces of soda, sports drinks, fruit drinks, and sweetened milks. Test‐retest reliability was measured (r=.65, p<.0001). Soda ‘times a week' (p=0.001) and sports drink 'times a day' (p=0.04) questions performed better than the soda ‘times a day' (p=.08). The 3‐item scale [recoded to the healthy direction] was negatively related to a sum of the volume of sweetened beverages on the food logs [r=0.31, p<0.01]. This pictorial assessment may be useful in assessing the child's sweetened beverage behaviors.Supported by National Research Initiative #2009‐55215‐05019 & Agriculture and food Research Initiative #2010‐85215‐20658 from the USDA National Institute of Food and Agriculture, Human Nutrition and Obesity 93330.
DETERMINANTS OF FRUIT AND VEGETABLE CONSUMPTION AMONG UNDERGRADUATE STUDENTS
Authors: Chairez‐Jimenez Carolina , Moreno‐Landa Luz Irene , Perez‐Lizaur Ana Bertha 
Published: June 2021

INTRODUCTION:Fruit and vegetables (FV) are core components of a correct diet, its consumption plays an important role in health and disease prevention. Including FV in diet is related to some factors, one of the most important is the stage of change where the individual is found.OBJECTIVE:Evaluate determinants in the consumption of FV among a group of undergraduate students.METHODS:Descriptive and transversal study. The group answered an on‐line questionnaire before and after their participation in a summer workshop. The sample was integrated for 78 students that were facilitators in the workshop “Jugando a Ganar Salud” in Mexico's State. A questionnaire to measure stage of change in FV consumption was used. The results were analyzed prior and post intervention. A Fisher test and Student “t” test were done.RESULTS:Age: 17‐24yrs,most of the participants studied health related careers (67.5%), in private universities (67.5%). A significant increase (p=0.018) was observed in search of information about how to increase consumption of FV, and in the perception of supermarkets to make it easy to buy FV (p=0.048). There was observed an increase in social liberation (p=0.0503), and a decrease in self‐liberation (p=0.0431). The most common processes of change were:consciousness raising, environmental reevaluation, self‐reevaluation, self‐liberation and reinforcement management.CONCLUSIONThere was a perception of availability; nevertheless, there is a lack of self‐efficacy. The results show the necessity of increase self‐efficacy in youth,so they can be capable to make healthy choices. Interventions in this age group are valuable because many habits generated during youth,persist to adulthood.
Milk vs Soybean Polar Lipids as Emulsifier: Faster Postprandial Lipid Metabolism through Greater Emulsion Lipolysis
Authors: Lecomte Manon , Bourlieu Claire , Cheillan David , Geloen Alain , Laugerette Fabienne , Michalski Marie‐Caroline 
Published: June 2021

ObjectiveBecause metabolic diseases are characterized by an altered lipid metabolism including prolonged postprandial hyperlipemia, we investigated the impact of using milk polar lipids as emulsifier on lipid digestion and postprandial lipemia.Methods:Mice were gavaged with emulsions stabilized with polar lipids from milk (MPL) vs soybean (SPL). Plasma and small intestine were collected during digestion. Both emulsions were also lipolyzed in vitro using a static human digestion model.Results:In vivo, MPL induced higher plasma concentrations of triglycerides (TG) and unesterified fatty acids (NEFA) than SPL at 1h after feeding. The opposite was observed after 4 h with lower TG in MPL group (PPLxtime<0.01). This was associated with a lower gene expression of ApoB and Sar1B, which are proteins involved in chylomicron structure and secretion, in the duodenum of MPL vs SPL mice (P<0.05). Chylomicrons were larger in MPL group at 4 h (P<0.01). The kinetics of synthesis and/or clearance of chylomicrons thus depended on the type of emulsifier. In vitro, TG hydrolysis from MPL emulsion was faster during intestinal phase than using SPL. This enhanced lipolysis can partly explain the sharper rise of plasma TG at 1 h in MPL group and its subsequent faster decrease.ConclusionsMilk polar lipids used as emulsifier can modulate both digestive lipolysis and postprandial lipid metabolism compared to soybean lecithin. Possible long‐term effects in humans will thus deserve further investigations.
Fruit and Vegetable Consumption is Inversely Related to Mild Cognitive Impairment among Older Adults
Authors: Kim Sunghee , Yu Areum , Choi Bo Youl , Kim Mi Kyung , Nam Jung Hyun , Oh Dong Hoon , Yang Yoon Jung 
Published: June 2021

Objective:This study was conducted to investigate the association between fruit and/or vegetable consumption with the risk of mild cognitive impairment (MCI) in Korean older adults.Methods:Subjects were 729 (307 men and 422 women) older adults aged 60 years and older participating in the Yangpyeong Cohort Study between 2011 and 2012. Dietary intake was estimated by using a food frequency questionnaire. Cognitive function was evaluated by Korean version of Mini‐Mental State Examination for Dementia Screening (MMSE‐DS). Multiple logistic regression analysis was applied to determine the associations between ‘Fruit', ‘Vegetable' and ‘Fruit and Vegetable' consumptions and the risk of MCI.Results:Age, depression score, proportions of current drinker and smoker decreased across the quintiles of ‘Fruit', ‘Vegetable' and ‘Fruit and Vegetable', but body mass index (BMI), total energy intake, fish consumption, proportion of higher education level and dietary supplement user increased across the quintiles of ‘Fruit', ‘Vegetable' and ‘Fruit and Vegetable'. ‘Fruit' (5th v. 1st quartile, OR = 0.46, 95% CI = 0.24‐0.88, p for trend = 0.081) and ‘Fruit and Vegetable' (5th v. 1st quartile, OR = 0.42, 95% CI = 0.21‐0.84, p for trend = 0.029) intakes were inversely related to the risk of MCI after adjusting for potential confounder.ConclusionsThe present findings suggest that high intakes of fruits and vegetables, but not vegetable, may have a beneficial effect on the prevention of MCI.