bluuebunny/crossref_metadata_2025_split huggingface dataset:
Leucyl‐tRNA synthetase is an amino acid sensor for the activation of Vps34 upstream of mTORC1 (1012.5)
Authors: Yoon Mee Sup , Chen Jie 
Published: June 2021

Amino acids availability regulates the mammalian target of rapamycin (mTOR) complex, mTORC1, a master regulator of cell growth. We previously reported that a pathway consisting of the class III PI‐3‐kinase Vps34 and the phospholipase PLD1 mediates amino acid activation of mTORC1, via PLD1 activation and translocation to the lysosome. The mechanism by which the Vps34‐PLD1 pathway senses amino acids is not known. In the current study, we identify leucyl‐tRNA synthetase (LRS) as the amino acid sensor for the activation of Vps34‐PLD1. LRS knockdown impaired amino acid‐induced Vps34 activation, cellular PI3P level increase, PLD1 activation, and PLD1 lysosomal translocation. Conversely, overexpression of LRS enhances activation of Vps34 and PLD by amino acids. Results with LRS mutants suggest that this function of LRS is dependent on its ability to bind leucine but independent of its tRNA charging activity. In addition, we find that LRS physically interacts with Vps34 through Vps15 and this interaction may be directly responsible for the activation of Vps34 in response to amino acids. Taken together, these observations strongly suggest that LRS mediates amino acid activation of Vps34‐PLD1 upstream of mTORC1. Our findings reveal a new regulator for Vps34 and fill a previous gap in the amino acid‐sensing signaling network.Grant Funding Source: NIH GM089771 and AR048914
NAADP evokes calcium signals and plays a critical role in downstream physiological responses in naïve T cells (1013.5)
Authors: Ali Ramadan , Camick Christina , Wiles Katherine , Slama James , Giovannucci David , Wall Katherine 
Published: June 2021

Nicotinic acid adenine dinucleotide phosphate (NAADP) is the most potent Ca2+ mobilizing second messenger discovered to date and has been implicated in Ca2+ signaling in human Jurkat lymphoma T cells via ryanodine receptor activation. In contrast the role of NAADP in Ca2+ signaling or the identity of the Ca2+ stores targeted by NAADP in conventional naïve T cells has not been investigated. In the current study, we demonstrated for the first time to our knowledge the importance of NAADP in the generation of Ca2+ signals in murine conventional naïve T cells. Combining live‐cell imaging methods and a pharmacological approach using the NAADP antagonist Ned‐19, we addressed the impact of NAADP on the generation of Ca2+ signals evoked by TCR stimulation and the role of this signal in downstream physiological endpoints such as proliferation, cytokine production, and other responses to stimulation of murine naïve T cells. Moreover, we demonstrated that acidic stores in addition to the endoplasmic reticulum are the relevant Ca2+ stores that were sensitive to NAADP in naïve T cells.Grant Funding Source: Supported by NIH GM100444
Microarray analysis of the effects of elevated cAMP in differentiating osteoblasts (1013.6)
Authors: Ghanayem Kathryn , Cuppini Candice , Rylaarsdam Robin 
Published: June 2021

McCune‐Albright Syndrome (MAS) is an acquired genetic disorder caused by a mutation in Gs alpha at Arg201 that inhibits GTP hydrolysis, constitutively activating the protein and resulting in elevated levels of basal cAMP in the affected cells. A major pathological feature of MAS is polyostotic fibrous dysplasia of bone (FD), a finding for which very little effective therapy exists. In an effort to identify novel potential drug targets to treat FD, we utilized a human fetal osteoblast cell line, hFOB1.19 (ATCC) that proliferates at 33C and differentiates into mature osteoblasts at 39C. We treated cells with the fungal metabolite forskolin at its EC50 concentration for elevating cAMP in these cells, and then either maintained the cells at 33C or shifted them to the differentiation conditions for 24 hours. Total RNA was collected and analyzed for levels of gene expression using an Illumina HT12 bead chip. Pairwise analysis of changes in gene expression identified 370 genes that were aberrantly expressed during differentiation in the presence of elevated cAMP. Twenty‐eight genes with known functions in osteoblast biology and/or differentiation were further analyzed by RT‐PCR to confirm the expression changes seen in the microarray assay.Grant Funding Source: Supported by NIH grant 1R15ED020190‐01
Cultural capital associated with children’s food consumption and physical activity among parents of first and second grade children in urban, suburban, and rural Japan (1014.1)
Authors: Goto Keiko , Fukushima Naho , Wolff Cindy , Giovanni Maria , Murayama Nobuko 
Published: June 2021

This study examined children’s food consumption and physical activity and their parents’ cultural capital levels in three Japanese communities. A total of 729 parents of first and second grade children enrolled in nine elementary schools in Chiba (urban), Niigata (suburban), and Miyazaki (rural) participated in a survey. Factor analysis resulted in three cultural capital variables: preservation of traditional Japanese food culture, parents’ aversion to Western foods, and parents’ openness to foods from other cultural groups. Correlations and ANOVA were used for statistical analysis. Children in Chiba consumed significantly higher amounts of store bought vegetables, fruits, and Western franchise fast foods compared to children in Miyazaki. Consumption of local vegetables was the highest in Miyazaki. Preservation of traditional Japanese food culture was associated with a higher level of physical activity among children. Western food aversion was associated with consumption of fresh vegetables and local/seasonal foods. Openness to trying foods from other cultures was associated with consumption of sweetened beverages. Our findings underline the significance of identifying cultural capitals for child health. These may ultimately contribute to developing policies and programs for healthier feeding practices in countries that are undergoing nutrition transition.Grant Funding Source: CSU Research grant
Global trends in breastfeeding duration (1015.1)
Authors: Kay Melissa , Bentley Margaret , Adair Linda 
Published: June 2021

Breastfeeding is an important determinant of child survival and strategy for improving child health in developing countries. The World Health Organization recommends exclusive breastfeeding up to 6 months, with continued breastfeeding along with appropriate complementary foods up to 2 years or beyond.We analyzed nationally representative Demographic and Health Survey (DHS) data from 21 countries between 1990 and 2011. Countries had at least 2 years of data, including one from 2008 or later for a total of 83 surveys. Our sample includes singleton, last‐born infants 0‐36 months living with their mother (n=185,469). We compared median breastfeeding duration calculated from current status data.Trends within countries did not exhibit meaningful patterns. Median duration changed by <1 month in 43% of intervals. Significant differences are seen between countries; 14% of country‐years had medians 2 years.Despite progress promoting awareness of breastfeeding, adherence to recommendations is not common. Analyzing DHS data is a unique opportunity to assess trends and explore noncompliance. These observations can be used to guide promotion efforts. Future studies should exam the influence of social, economic and demographic characteristics on breastfeeding duration.Grant Funding Source: Supported by an educational grant from Wyeth Nutrition
Consumption of low‐fat dairy foods for 6 months improves insulin resistance without adversely affecting lipids or bodyweight in healthy adults (1025.2)
Authors: Rideout Todd , Marinangeli Christopher , Rempel Curtis 
Published: June 2021

Given the highly debated role of dairy food consumption in modulating biomarkers of metabolic syndrome, this study was conducted to examine the influence of long‐term (6 month) dairy consumption on metabolic parameters in healthy volunteers under free‐living conditions without energy restriction. Twenty‐three healthy subjects completed a randomized, crossover trial of 12 months. Participants consumed their habitual diets and were randomly assigned to one of two treatment groups: a high dairy supplemented group instructed to consume 4 servings of dairy per day (HD); or a low dairy supplemented group limited to no more than 2 servings of dairy per day (LD). Baseline, midpoint, and endpoint metabolic responses were examined. Endpoint measurements of body weight and composition, energy expenditure, blood pressure, blood glucose, and blood lipid and lipoprotein responses did not differ (p>0.05) between the LD and HD groups. HD consumption improved (p<0.05) plasma insulin (‐9%) and insulin resistance (‐11%, p=0.03) as estimated by HOMA‐IR compared with the LD group. Study results suggest that high dairy consumption (4 servings/d) may improve insulin resistance without negatively impacting bodyweight or lipid status under free‐living conditions.Grant Funding Source: Supported by the Government of Manitoba Department of Industry, Energy and Mines
Soda consumption and the risk of hip fractures in postmenopausal women (1027.6)
Authors: Fung Teresa , Arasaratnam Meredith , Grodstein Francine , Katz Jeffrey , Rosner Bernard , Willett Walter , Feskanich Diane 
Published: June 2021

Background: The frequency of soda consumption remains high in the U.S. Soda consumption has been associated with poor bone health in children, but few studies have examined this relationship among adults, and none have examined the relation of soda consumption to risk of hip fractures.Objective: We examined the association of soda, including specific types of soda, and risk of hip fracture in postmenopausal women.Design: Analysis was conducted using data from the ongoing Nurses’ Health Study cohort. Diet was assessed at baseline using a semi‐quantitative food frequency questionnaire, and updated approximately every four years. In up to 30 years of follow‐up, we identified 1,968 incident hip fractures in postmenopausal women (N=76,123). We computed relative risks (RR) for hip fracture by amount of soda consumption using Cox proportional hazards models, adjusting for potential confounders.Results: In multivariable models, each additional serving of total soda per day was associated with a significant 14% increased risk of hip fracture (RR=1.14, 95% confidence interval (CI) 1.07‐1.23). Risk was significantly elevated among consumers of both regular (RR=1.19, 95% CI= 1.03‐1.38) and diet soda (RR=1.12, 95% CI= 1.04‐1.21) and also did not significantly differ between colas and non‐colas or sodas with or without caffeine. Association between soda and hip fractures did not differ by body mass index or by diagnosis of diabetes.Conclusion: Increasing soda consumption of all types may be associated with increasing risk of hip fracture in postmenopausal women.Grant Funding Source: Supported by: NIAMS T32 AR 055885; NIAMS P60 AR 47782, NIH CA87969
Interaction of the protease resistant proteins with the intestinal mucosa membrane proteins (1036.3)
Authors: Yum Youjin , Lee Juyoung , Lim Jinkyu 
Published: June 2021

Many studies have shown that some substances in soy, such as digested peptides or isoflavones from soy have effects on satiety control and nutrient absorption functions. However, it is still clearly not known how soy can control those functions. We have shown that some of the soluble soy proteins were resistant to proteases, such as pepsin and pancreatine, in the in vitro digestion analysis. We named these proteins as PRFPs (protease‐resistant food proteins), and postulated that PRFPs can interact with the mucosa membrane on the villi or microvilli in the small intestine to trigger the biological activities mentioned above. The interactor proteins on the intestinal membrane and soy proteins were identified by crosslinking and mass spectrometric analysis. The identified intestinal membrane proteins were mostly channel proteins and galectins. The soy partners to the membrane proteins were found to be glycinin and beta‐conglycinin, which are the majority proteins of soy. Phosphoproteomic changes in the intestinal cell line upon soy protein treatment were tested. After the soy protein treatment, phosphoproteins enriched from the cell extract by IMAC column were separated on 2D gels and compared for the up and down regulations. Interestingly, many of the proteins involved in protein folding or energy metabolism were significantly phosphorylated by soy protein treatment to the cell line.For further studies, detailed cell signaling pathway/mechanism by soy PRFPs treatment to the cells are under investigation in view of revealing the function of soy proteins in satiety, nutrient absorption, and others.
γ‐Conglutin, the lupin seed glucose‐lowering glycoprotein, interferes on insulin pathway in Hep G2 cells (1045.16)
Authors: Parolari Anna , Manzoni Cristina , Faoro Franco , Duranti Marcello , Lovati Maria 
Published: June 2021

Gamma‐conglutin (γ‐C), a Lupinus albus, seed glycoprotein, has been shown to lower blood glucose in hyperglycaemic rats and increase glucose consumption in HepG2 cells. Aim of the present work was to assess the modalities of interaction of this protein with the target cells, so as to contribute unveiling its mechanism of action. γ‐Conglutin internalization: HepG2 cells, pretreated for 30’ w/wo inhibitors of caveolae mediated endocytosis [filipin (5 μg/ml) and genistein (200 μM)], clathrin‐mediated pathway [chlorpromazine (25 μM), methyl‐β‐cyclodextrin (5 mM)], and macropinocytosis [amiloride (5 mM)], were incubated with FICT‐γ‐conglutin (50 μg/ml) for 4 h at 37°C, followed by FACS analysis. The macropinocitosis pathway was the mechanism of γ‐conglutin internalization (69% inhibition). 2D‐electrophoresis of cell lysates and antibody reaction of the blotted maps showed that γ‐conglutin is internalized in intact form. Moreover, the protein is modified inside the cell by multiple phosphorylation. Insulin pathway: reduced effect on AMPK activity (‐50% vs controls) and increased expression of phosphorylated AKT form (+ 25% vs controls) were recorded in HepG2 cells, following western‐blot analysis, exposed to 10 mM γ‐conglutin and/or 100 nM insulin, or metformin 10 mM.Grant Funding Source: MIUR of Italy
cDNA and lincRNA microarrays: identification of expression patterns related to metastatization in oral squamous cell carcinoma (1048.6)
Authors: Oliveira‐Costa Joao Paulo , Gigliola Mabel , Macedo Mariana , Carraro Dirce , Soares Fernando 
Published: June 2021

Oral squamous cell carcinoma (OSCC) metastatic cascade still a poorly understood mechanism, in terms of gene expression differences between primary site and metastatic cells at lymph nodes. Using 20 matched samples of OSCC cases, being 10 from primary sites and 10 metastastic lymph nodes we performed lincRNA and cDNA microarrays analysis (Agilent`s Sureprint G3 Human Genome V2.0). Our results showed 617 genes and lincRNAs significantly differentially expressed between metastatic lymph nodes and primary tumors. Integrated pathway analysis showed alteration in canonical pathways such as Ras homolog gene family, member A signaling pathway (RhoA pathway; p=0.002) and tumor necrosis factor receptor 2 signaling pathway (TNFR2 pathway; p=0.009). The most up‐regulated genes included SNORA65 (9,045‐fold), ZMAT1 (4,776‐fold) HNRNPU‐AS1 (4,626‐fold), RNU12 (4,557‐fold), TLE4 (4,555‐fold), and the most down‐regulated included MYBPH (‐67,818‐fold), ACTA1 (‐58,551‐fold), TNNC1 (‐23,119‐fold), F2RL2 (‐17,431‐fold) and MYLPF (‐13,785‐fold). Regarding lincRNAs, MALAT1 was significantly overexpressed in metastatic lymph nodes, with a 4.2‐fold increase. Our study shows the existence of several genes and lincRNAs with different expression patterns between OSCC primary tumor and metastatic lymph nodes, and also points to novel genes, which has not yet been implicated in OSCC metastasis, providing new potential targets in oral cancer research.Grant Funding Source: Supported by FAPESP grants 2011/18606‐7‐0 and 2010/08400‐0
Cooperative gating and sensitivity of TRPV4 channels are regulated by distinct factors (1057.8)
Authors: Sonkusare Swapnil , Villalba Nuria , Freeman Kalev , Bonev Adrian , Nelson Mark 
Published: June 2021

TRPV4 (transient receptor vanilloid 4) sparklets represent Ca2+ influx through single TRPV4 channels (Sonkusare et.al., Science, 2012). In this study, we determined the differences in endothelial cell (EC) TRPV4 channel activity in mouse mesenteric (MA) and basilar (BA) arteries by recording TRPV4 sparklet activity in slit open, en face preparation, under physiological conditions of temperature and ionic solutions. TRPV4 channels in MAs and BAs exhibited a number of similarities, including a 4‐channel meta‐structure, the same quantal levels (0.19 ΔF/F0), and cooperative channel gating at myoendothelial projections (MEPs). However, an analysis of TRPV4 sparklets indicated that the channel open probability (Po) in response to TRPV4 agonist GSK1016790A (GSK101, 100 nM) is ~ 15‐fold lower in BAs than MAs. In agreement with the Ca2+ imaging results, the sensitivity of TRPV4 channels to the agonist GSK101, measured as currents in freshly isolated cells is shifted by ~10‐fold in cerebral arteries. The TRPV4 channel gating at MEPs was cooperative in both MAs and BAs, as indicated by similar coupling coefficients (~0.2). In MAs, the cooperative channel gating at the MEPs was dependent on a kinase anchoring protein 150 (AKAP150). Notably, AKAP150 exhibited similar clustering at MEPs in MAs and BAs. These results suggest that cooperativity and agonist sensitivity of TRPV4 channels are functionally separable.Grant Funding Source: HL044455, 1P01HL095488, R37DK053832, R01HL098243 to MTN
Analysis of functional selectivity at the nociceptin opioid receptor (1066.2)
Authors: Chang Steven , Spangler Skylar , Zhang Nancy , Planer William , Carroll F , Bruchas Michael 
Published: June 2021

The Nociceptin/Orphanin FQ Opioid Receptor (NOPR) is the most recently discovered and least characterized G‐protein coupled receptor in the opioid receptor family. The activity of NOPR has been thought to play a role in neuromodulation in the contexts of pain, opioid tolerance, and the modulation of stress and anxiety behaviors. It is thought that the NOP receptor may bind functionally selective ligands which can bias signal transduction, resulting in multiple behavioral manifestations. Functional selectivity at the NOPR has yet to be explored, but this information is integral to understanding the complex modulation of behaviors. To study the functional selectivity of NOPR, we used live‐cell cAMP assays, BRET, as well as MAP kinase activity assays to study the Gα‐protein mediated and arrestin‐mediated signaling profiles. We screened multiple known NOPR ligands and novel, small molecules aimed at elucidating structural significance. We have found that NOPR does exhibit different signaling profiles, depending on the interacting ligand. Specifically, we show that minute changes in the structure can illicit opposing signaling profiles ranging from agonism to inverse agonism. Additionally, we propose ligand‐receptor docking configurations that may help elucidate the conformational influence on functional selectivity. Additionally, we screened the same ligands in an internalization and arrestin‐recruitment deficient NOPR point mutant (Zhang et al., 2012). Interestingly, the mutation of the Serine 363 phosphorylation site shows altered signaling profiles for the same ligands in comparison to the wild‐type receptor. In addition, the signaling profiles of the novel NOPR ligands may provide new insight into the structural relationship between ligands and biased signal transduction. Taken together, these data provide novel insight into the functional selectivity of the NOP receptor, and how it affects behavioral manifestation. Supported by: NIH R00 DA025182 and T32 DA007261.Grant Funding Source: T32 DA007261
Calcium/calmodulin‐dependent protein kinase II modulates inositol‐3‐phosphate receptors in vascular endothelium from mouse mesenteric arteries (1075.1)
Authors: Toussaint Fanny , Charbel Chimène , Blanchette Alexandre , Comtois Philippe , Ledoux Jonathan 
Published: June 2021

Endothelial Ca2+ pulsar consists in a spontaneous Ca2+ release from IP3R localized within myoendothelial projections (MEP). We recently showed CaMKII activation by Ca2+ pulsars, thus modulating endothelial function through NO production. Moreover, CaMKII is a known regulator of intracellular Ca2+ homeostasis in several cell types. Our hypothesis was that Ca2+ pulsar‐activated CaMKII modulates endothelial intracellular Ca2+ dynamics. Our investigation showed that preincubation with KN‐93 (10 µM), CaMKII inhibitor, significantly altered endoplasmic reticulum (ER) Ca2+ content (≍ ‐60%). A similar KN‐93‐induced alteration of ER Ca2+ levels (≍ ‐55%) was observed while SERCA pumps were inhibited (Thapsigargin; 1 µM). However, inhibition of IP3R (2APB; 100 µM) precluded KN‐93 effect on ER Ca2+ content. Colocalization (<40nm) of CaMKII and IP3R was observed by PLA in situ hybridization. In addition, an isoform‐specific distribution within MEP (IP3R2>IP3R1>IP3R3) has been found by in situ immunofluorescence. Finally, KN‐93 induced recruitment of new Ca2+ pulsars sites (1.8‐Fold). These results suggest that CaMKII modulates intracellular Ca2+ dynamics through modulation of IP3R activity. Thereby, CaMKII is involved in a negative feedback loop as an endothelial calcium‐sensor switch regulating Ca2+ homeostasis.Grant Funding Source: Supported by FRQS, FICM, CFI, SQHA and HSFC
Sex specific effect of antenatal glucocorticoids on nitric oxide production stimulated by angiotensin peptides in renal proximal tubule cells (1085.6)
Authors: Bi Jianli , Chen Kai , Su Yixin , Chappell Mark , Rose James 
Published: June 2021

We have shown that renal proximal tubule cells (RPTCs) from male offspring exposed to antenatal betamethasone (B) increase Na+ uptake during Angiotensin II (ANG II) stimulation more than RPTCs from vehicle exposed animals while Na+ uptake by cells from females is not altered by B. The ability of angiotensin 1‐7(ANG 1‐7) to decrease sodium uptake was blocked in RPTCs from males but not in RPTCs from females exposed to B . Sodium uptake may relate to nitric oxide (NO) production in RPTCs. Therefore, this study aimed to assess the effect of antenatal glucocorticoid on nitric oxide production stimulated by angiotensin peptides in RPTCs in adult sheep exposed to B.We obtained kidneys from 1‐1.5 year old male and female sheep exposed to either B (n=5) or Vehicle (V) (n=5) at 80‐81 days gestation. RPTCs were cultured in DMEM/F12 and stimulated with ANG II or ANG1‐7 with or without blockers. Nitric oxide production in cells was measured by DAF‐FM diacetate, nitric oxide indicator. ANOVA was used for data analysis.ANG II suppressed NO production and the suppression was blocked by candesartan (in males: F=4.2, P=0.05; in females: F=5.9, P=0.02). In presence of PD the suppression was greater in male antenatal B than male V exposure sheep (F=17.8, P=0.006). Compared with females, suppression of NO production by ANG II is greater in male B exposed sheep (F=10.7, P=0.005). Increase of NO production by ANG 1‐7 was greater in RPTCs in male V than B treated sheep (F=5.5, P=0.03). There was no difference in the increase of NO production by ANG 1‐7 in female B vs. V sheep. The increase was blocked by D‐Ala. NO production by ANG 1‐7 was greater in female than male B exposed sheep (P=0.02). The NO production was blocked by L‐NAME. The results suggest antenatal B alters NO changes in response to ANG II and ANG 1‐7 in male but not in female RPTCs. The suppression of NO production by ANG II is mediated by AT1 receptor and NO production induced by ANG 1‐7 may involve in Mas receptor. The differences in NO may mediate sex related differences in Na+ uptake by these cells.Grant Funding Source: Supported by HD‐47584 and HD‐17644.