bluuebunny/crossref_metadata_2025_split huggingface dataset:
Vesicular release of ATP by brainstem astrocytes evoked by changes in pH
Authors: Kasymov Vitaliy , Marina N , Kasparov S , Gourine A 
Published: June 2021

The mechanisms of central chemosensory function are not completely understood. Here we explored the mechanisms underlying release of ATP by brainstem astrocytes in response to changes in pH.RESULTSLoading of astrocytes with fluorescent ATP analogue MANT‐ATP resulted in abundant punctate fluorescent staining in the cytosol and this was prevented by bafilomycin A1, indicating vesicular accumulation of the marker. In brainstem astrocytes, TIRF imaging revealed progressive decreases in MANT‐ATP fluorescence intensity in response to a decrease in external pH indicating fusion of the labelled compartments. The number of fusion events triggered by acidification in brainstem astrocytes loaded with BAPTA‐AM was significantly reduced indicating that vesicular fusion requires intracellular Ca2+ signalling. The fusion rate in cortical astrocytes was not affected by acidification. Acidification‐evoked vesicular fusion responses in brainstem astrocytes were unaffected by blockade of ATP‐mediated signalling with MRS2179 or apyrase. Quantitative RT‐PCR analysis of genes encoding proteins associated with vesicular fusion revealed that expression levels of VAMP3, Slc1a2 and Slc17a9 are significantly higher in brainstem astrocytes as compared to cortical astrocytes.CONCLUSIONSChemosensory stimulation of brainstem astrocytes triggers vesicular fusion underlying release of ATP.
Adaptive Exploitation of Stromal Cell Metabolism by Tumor Cells
Authors: Patel Brij , Rattigan Yanique , Ackerstaff Ellen , Koutcher Jason , Sukenick George , Glod John , Banerjee Debabrata 
Published: June 2021

BACKGROUND/OBJECTIVEGlycolytic tumors secrete lactate into the tumor microenvironment (TME). Glycolytic muscle cells also secrete lactate, while neighboring oxidative muscle cells use this lactate as an energy source. We propose that lactate mediates tumor‐TME metabolic coupling similar to that of muscle cells. Using in vitro‐derived activated TME cells (CAFs), we investigated the role that lactate may play in CAF‐mediated support of tumor growth.METHODS/RESULTS[1] We find that MDA‐MB‐231 (MDA) breast cancer cells secrete 3‐fold more lactate under hypoxia, and that lactate recruits CAF precursors towards MDAs. [2] Lactate is transported by monocarboxylate transporters (MCTs); it is taken up via MCT1 and effluxed via MCT4. MDAs have low MCT1 but high MCT4 expression, while CAFs show high MCT1 but low MCT4 expression, implying that tumor‐extruded lactate is taken up by CAFs. [3] 13C NMR spectroscopic analyses indicate that 13C‐lactate is indeed metabolized via the Krebs cycle in CAFs. This implies that CAFs oxidatively metabolize lactate. [4] MDAs co‐cultured with CAFs display greater uptake of exogenous pyruvate, and subsequent 13C‐pyruvate tracking assays indicate the presence of 13C‐labeled lactate. Collectively this suggests that tumor cells may derive sources of energy from lactate‐oxidizing components of their TME. [5] Ongoing studies investigating MCT1 blockers CHC and Leutolin indicate that inhibiting lactate transport in CAFs may inhibit tumor growth.CONCLUSIONSTo our knowledge this is the first in vitro model system demonstrating tumor/TME metabolic coupling by which tumor cells exploit stromal cells. A better understanding of the molecular mechanisms governing metabolic cooperation within the tumor milieu will potentially identify new targets for therapeutic intervention. Grant Funding Source: NIH P50 CA86438 & NJCCR‐10‐1964‐CCR‐EOGrant Funding Source : NJCCR‐10‐1964‐CCR‐EO
Simple Western analysis of NFκB signaling cascade proteins
Authors: Boge Annegret , Nguyen Uyen , Ramirez Francisco , Xu Hui , Kazakova Irina , Yang Tom , Gavin Robert 
Published: June 2021

Aberrant expression in the NFκB pathway is a common occurrence in inflammatory and stress‐induced diseases such as cancer. Understanding how NFκB signaling impacts disease progression is important to the development of novel therapeutics. Western blots, while semi‐quantitative and tedious to perform are commonly used. The Simple Western™ platform, provides a solution that enables the complete automation of traditional Westerns allowing analysis of up to 96 samples in a single experiment. Simple Westerns use nanoliter volumes, enabling the characterization of precious or limited quantity samples providing reproducible and quantitative results.Targets from the NFκB pathway, including IκB, NFκB subunits c‐ Rel, p65, p50/p105 along with loading controls were run on the Simple Western platform. In response to TNF‐α treatment, statistically significant changes of key target proteins were characterized.The Simple Western permits quantitative high throughput analysis of signaling pathways at a level of efficiency and reproducibility not obtainable by traditional Westerns, allowing better characterization of regulatory events and key biomarker targets involved in the progression of disease states.
Study of the Compartmentalization of Type 1 Angiotensin Receptor Using Bioluminescence Resonance Energy Transfer‐based Sensors
Authors: Hunyady László , Soltész-Katona Eszter , Várnai Péter , Balla András 
Published: June 2021

Initiation and termination of signaling of the type I angiotensin receptor (AT1‐R) can lead to dynamic changes in its localization in membrane microdomains.We used YFP‐labeled fusion constructs constructs (i.e. raft‐, or non‐raft plasma membrane markers) to analyze the molecular dynamics of AT1‐R trafficking in response to stimulation in bioluminescence resonance energy transfer (BRET) measurements. Our data demonstrate that angiotensin II (AngII) stimulus changes the microdomain localization of wild type or mutant (DRY/AAY or TSTS/A) AT1‐Rs in HEK293 cells. The comparison of the trafficking of AT1‐R upon AngII stimulus to either [Sar1,Ile8]‐ AngII or [Sar1,Ile4,Ile8]‐AngII stimulus revealed different type of molecular dynamics depending on the nature of the ligand of the AT1‐R. The molecular dynamics of AT1‐R also changes upon cholesterol depletion, which suggests that the mechanism is raft‐dependent. The molecular dynamics of the AT1‐R are strikingly differ from those of 5HT‐2C and EGF receptors, which demonstrate the usefulness of the BRET based measurements in the investigation of receptor trafficking in living cell experiments. These studies should provide valuable information about the distribution and molecular dynamics of AT1‐R upon angiotensin stimulus among membrane microdomains.Supported by Hungarian National Grant OTKA NK‐100883, TÁMOP 4.2.1 B‐09/1/KMR‐2010‐0001.
Spatial Analyses of Healthy Eating Index Reveal a Relation between Diet Quality and Place
Authors: Aggarwal Anju , Moudon Anne Vernez , Cook Andrea , Drewnowski Adam 
Published: June 2021

The Healthy Eating Index (HEI 2005) is a measure of diet quality, based on compliance with federal recommendations and guidelines. Higher HEI values have been consistently linked with higher education and incomes. However, diet quality measures still remain to be linked to specific geographic locations or neighborhoods. The Seattle Obesity Study (SOS) was a study in spatial epidemiology that collected data on home and work locations, socioeconomic status (SES), diets and health for 1,323 adult residents’ of King County, WA using a combination of telephone surveys and mailed food frequency questionnaires. Home addresses of study respondents were geocoded. Residential property values were obtained from the King County assessor tax parcel database. Multivariate regressions examined associations between HEI scores and property‐based novel metrics of personal and neighborhood SES. Higher property values were associated with significantly higher HEI 2005 scores, even after adjusting for individual level SES and demographic characteristics. Geographic Information Systems (GIS) approaches permit new spatial research on diet quality in relation to neighborhood resources and food access. These findings are the first to link HEI values, a measure of diet quality, to features of the built environment at the individual level.Supported by NIH grants R01 DK076608 and R21 DK 085406
Contribution of flavoured milk on dietary intakes and its association with body mass index percentile in children at high risk of overweight
Authors: Wang JiaWei , Shang Lei , Johnson-Down Louise , St-Arnaud-McKenzie Danielle , Lambert Maire , Paradis Gilles , Gray-Donald Katherine 
Published: June 2021

To compare children's dietary intake and body mass index percentile (BMI%ile) between those who did and did not consume flavored milk (FM). In the Québec Adiposity and Lifestyle Investigation in Youth (QUALITY) study (2005–2008), 632 Caucasian children aged 8–10 years (with at least one obese parent) were recruited. Assessment of FM, sugar sweetened beverage (SSB) and nutrient intakes was based on three 24‐hour recalls. Height and weight were measured for calculating BMI%ile. Physical activity was scaled using 7‐day accelerometer. Time spent in the sedentary behavior was collected using standard questionnaire. The percentage of overweight children was 41.9%. FM drinkers (N = 90) consumed 132g/d FM per day and had a higher mean intake for calcium (930 vs. 837 mg; P = 0.010), Vitamin D (6.9 vs. 5.9 μg; P = 0.021) and total sugar (99 g vs. 90 g, P = 0.015) than non‐FM drinkers (N = 521). These differences remained significant after adjustment for total energy intake. In multivariate analysis controlling for physical activity, sedentary behavior and SSB intake, FM consumption was not associated with BMI%ile. FM consumption was not associated with BMI%ile despite higher sugar intake and FM provided higher levels of calcium and Vitamin D intake.Funding: Canadian Institutes of Health Research, Heart and Stroke Foundation of Canada, Fonds de la recherche en santé du Québec and Dairy Farmers of Canada
Acai Juice with Baker's Yeast Beta‐Glucan Reduces URTI Symptoms and Improves Mood State in Stressed Subjects
Authors: Talbott Shawn , Talbott Julie , Vosti Mark , Blackhurst Darin 
Published: June 2021

BACKGROUNDSeveral studies have shown Baker's yeast Beta‐1,3/1,6 D‐glucan, extracted from Saccharomyces cerevisiae is effective in reducing the incidence of cold and flu symptoms. Other studies have shown acai berry juice to provide antioxidant benefits and reduce oxidative damage.OBJECTIVEThis study evaluated the effect of a combination of acai berry and Baker's yeast beta‐glucan (MonaVie M‐Mun) on upper‐respiratory tract infection symptoms (URTIs) and physical/psychological well‐being in subjects with moderate levels of psychological stress.METHODSHealthy adults (41 ± 12y) self‐administered 4 ounces of Placebo juice (N=34) or M‐mun juice (N=45) daily for 12‐weeks. We used the Profile of Mood States (POMS) psychological survey and RAND SF‐36 Medical Outcomes Survey (MOS) to assess changes in mental/physical energy levels and overall well‐being. A quantitative health perception log was used to track URTI symptoms.RESULTSSubjects in the M‐mun group reported 43% fewer URTI symptoms compared to Placebo, as well as superior measures of physical & mental functioning, and overall well‐being (MOS +7%, POMS +6%).CONCLUSIONSThese data show that daily dietary supplementation with an acai/beta‐glucan juice (M‐mun) reduces URTI symptoms and improves mood state in stressed subjects, and thus may be a useful approach for maintaining immune protection against daily stressors.
Excessive autophagy contributes to neuron death in cerebral ischemia
Authors: Shi Ruoyang , Zhang Lin , Zhao Ling , Weng Jiequn , Li Xinmin , Kong Jiming 
Published: June 2021

AimsTo determine the extent to which autophagy contributes to neuronal death in cerebral hypoxia and ischemia.MethodsWe performed immunocytochemistry, western blot, cell viability assay, and electron microscopy to analyze autophagy activities in vitro and in vivo.ResultsIn both primary cortical neurons and SH‐SY5Y cells exposed to oxygen and glucose deprivation (OGD) for 6 h (hours) and reperfusion (RP) for 24, 48 and 72 h respectively, an increase of autophagy was observed as determined by the increased ratio of LC3‐II to LC3‐I and Beclin‐1 (BECN1) expression. Using Fluoro‐Jade C and monodansylcadaverine (MDC) double‐staining, and electron microscopy we found the increment in autophagy after OGD/RP was accompanied by increased autophagic cell death, and this increased cell death was inhibited by the specific autophagy inhibitor, 3‐Methyladenine (3‐MA). The presence of large autolysosomes and numerous autophagosomes in cortical neurons were confirmed by electron microscopy. Autophagy activities were increased dramatically in the ischemic brains 3 to 7 days post‐injury from a rat model of neonatal cerebral hypoxia/ischemia as shown by increased punctate LC3 staining and Beclin‐1 expression.ConclusionExcessive activation of autophagy contributes to neuronal death in cerebral ischemia. This study was supported by CIHR, MHRC, MICH and Canadian Stroke Network.
FOXO1 regulation of luteinizing hormone beta transcription
Authors: Skarra Danalea , Arriola David J. , Thackray Varykina 
Published: June 2021

Luteinizing hormone (LH) is required for mammalian fertility and is produced exclusively by gonadotrope cells within the anterior pituitary. Insulin stimulation of gonadotropes induces LH production, although the mechanisms are unknown. One candidate that may transduce insulin signaling into functional changes in Lhb synthesis is the Forkhead box‐O (FOXO) family of transcription factors. In response to insulin, FOXOs are negatively regulated by Akt phosphorylation, which results in their export from the nucleus to the cytoplasm, thereby limiting their nuclear activity. We have identified FOXO1 expression in murine pituitary gonadotropes and in LbT2 cells, an immortalized gonadotrope cell line. Here, we investigate insulin regulation FOXO1 and the ability of FOXO1 to alter Lhb synthesis. Western analyses identified phosphorylation changes in FOXO1 in response to insulin, which correlated with FOXO1 cytoplasmic localization using immunofluorescence imaging. By luciferase assay, we determined that FOXO1 overexpression in LbT2 cells inhibits basal and gonadotropin‐releasing hormone induced Lhb transcription. Our results indicate that FOXO1 is regulated by insulin in gonadotropes and FOXO1 inhibits Lhb synthesis. In summary, our findings identify a novel mechanism for Lhb gene regulation.Support: T31 HD720329, R01 HD067448, K01 DK080467, P30 DK063491, and U54 HD012303.
The effect of chromosome substitution on endurance exercise capacity in inbred and F2 generation mice
Authors: Courtney Sean Michael , Massett Michael P 
Published: June 2021

Previous research identified a region on chromosome 14 as an important regulator of endurance exercise capacity. The aim of this study was to identify the effect of chromosome substitution on endurance exercise capacity in inbred and hybrid mice. Mice from a chromosome substitution strain (CSS) derived from A/J (A) and C57Bl/6J (B6), denoted as B6.A14 were used to assess the contribution of chromosome 14 to endurance exercise capacity. To further investigate the heritability of the endurance exercise phenotype, 155 B6 x B6.A14 F2 mice were generated from a cross between B6 and B6.A14 CSS mice. All mice performed a graded exercise test to exhaustion to determine endurance exercise capacity. Endurance exercise capacity was expressed as time (min). B6 x B6.A14 F2 mice had an exercise capacity of 30.8 ± 0.1 minutes, which is significantly higher than parental B6.A14 CSS mice (28.5 ± 0.2 min) and similar to parental B6 mice (31.0 ± 0.1 min). Although the cohort mean is similar between F2 and B6 mice, the range of responses in the B6 x B6.A14 F2 mice (27.3 – 34.5 min) exceeded the range of the parental inbred strains. The range of responses in the B6 x B6.A14 F2 mice suggests that this population of mice is diverse in both phenotype and genotype and would be amenable to a chromosome‐wide scan for endurance exercise‐related genes.Supported by NIH grant HL085918 to MPM
Meal and snack patterns associated with biomarkers of glucose metabolism, diet quality and weight status in US adults
Authors: Shin Dayeon , Song Su Jin , Paik Hee‐Young , Song Won O. 
Published: June 2021

Epidemiological investigations established the associations between insulin resistance (IR), anthropometry and certain aspects of dietary intake. Our aim was to examine if meal and snack patterns are associated with biomarkers of glucose metabolism (fasting glucose and insulin, HbA1c, and HOMA‐IR), diet quality (HEI 2005), and weight (wt) status (BMI). Dietary recall data of 2175 men and 2191 women from the 2007–8 NHANES were subjected to factor analysis by gender by 19 food groups of MyPyramid Equivalents Database 2.0. Accounting for the complex survey design and sample wt, SAS v. 9.3 was used for logistic regressions. Of four meal patterns in men, two were associated (+) and one (−) with diet quality; the fourth(“grain and dairy”) was related with HbA1c and HOMA‐IR (p<0.05). Two meal patterns of women were associated (+) and one (−) with diet quality; the fourth (“fruit and alcohol”) indicated (+) for HbA1c, diet quality, and wt status. Snack patterns alone reflected total diet quality (p<0.05), but not biomarkers of glucose metabolism nor wt status in both gender. Meal and/or snack patterns of US adults are associated differentially with biomarkers of glucose metabolism, diet quality, and wt status.Grant Funding Source: None
Systematic review of Wild Rice: an ancient grain with modern benefits
Authors: Timm Derek Allen , Slavin Joanne 
Published: June 2021

Wild rice (Zizania sp.) grows in shallow waters of North America and is distinct from white rice (Oryza sp.). The recent trend toward gluten free foods and ancient grains presents an opportunity for increased interest in the health benefits of wild rice. Wild rice is classified as a whole grain and typically contains 74% starch, 6.8% dietary fiber, 14% protein, and 0.7–0.9% of lipids. Wild rice is a good source of dietary fiber and has a good protein efficiency ratio (1.7), which is high for a grain. The lipid content is low; however most of the lipids are Ω‐6 (35–37%) and Ω‐3 (20–31%) fatty acids with a very favorable ratio of (1.1). In addition to macronutrient content, wild rice contains phytosterols levels several times higher than other grains. Furthermore, the antioxidant activity of wild rice is between 10–15 times higher than white rice. A new study of wild rice added to a high fat/cholesterol diet showed significant attenuation of serum free fatty acids, total cholesterol, and triglyceride levels while maintaining higher high density lipoprotein levels compared to control. The exact explanation of the protective mechanism of wild rice is uncertain, but the dietary fiber, phytosterol, or antioxidant capacity of wild rice may be the reason for this. Current interest in ancient grains and gluten‐free diets, as well as antioxidants and phytochemicals, makes wild rice an attractive grain addition to the diet.Grant Funding Source: MN Wild Rice Council
NAD(P)H autofluorescence induction by Compound C in rat carotid chemoreceptor cells
Authors: Kim Insook , Fite Jaclyn Lasha , Kim Donghee , Donnelly David F. , Carroll John L. 
Published: June 2021

Compound C (CC) has been known as an inhibitor of AMP‐activated kinase (AMPK). Recently other roles of CC have been reported. CC inhibits hypoxia dependent HIF‐1α activation independent of AMPK but dependent on mitochondrial electron transport chain (ETC) (Emerling 07) and CC functions as an inhibitor of complex I of ETC (Chua 11). In addition, we have found that CC (4μM) produces ~10 fold increase in NAD(P)H autofluorescence (AutoFlu) as compared to hypoxia or cyanide in rat carotid body (CB) glomus cells. Autoflu was detected with 360/50 excitation(x) and 460/50 emission(m) filters. The effect of CC on the hypoxic [Ca2+]i response was observed by using 340&380x with 540/50m filter for fura‐2. Rat CB glomus cells were identified with rhodamine‐tagged peanut agglutinin.Results1) AutoFlu produced by CC is dose dependent, reaching a plateau at 0.04μM in both CB type cells. 2) The magnitude of AutoFlu induced by CC was greater in glomus cells compared to non‐glomus cells. 3) AutoFlu induced by 0.4μM CC was reduced ~50% by hypoxia but only < 10% by 20mM KCl. 4) CC did not increase calcium despite the increase in AutoFlu. 5) 0.4μM CC reduced the hypoxia‐induced [Ca2+]i increase by ~50%, but did not block 20mM KCl‐induced [Ca2+]i increase. These preliminary results suggest that CC induced significant NAD(P)H AutoFlu, which may significantly affect the measured [Ca2+]i response to hypoxia in glomus cells. (Funded by NIH HL54621)
Link between the prorenin/renin receptor and glucose homeostasis in mice fed a high‐fat/high‐carbohydrate diet
Authors: Shamansurova Zulaykho , Tan Paul , Bisotto Sandra , Michel Catherine , Nguyen Thi M‐D , Shiller Peter W , Gutkowska Jolanta , Lavoie Julie L 
Published: June 2021

ObjectiveElucidate the role of the (P)RR in glucose homeostasis in mice fed high‐fat/high‐carbohydrate diet (HF/HC).Material and methods12 week‐old C57BL/6 male mice were placed on a HF/HC or normal diet (ND) and were treated with either (P)RR blocker [(P)RR‐B] or saline for 10 weeks. Body weight was assessed weekly. At the end of treatment, mice were euthanized and fat tissues from subcutaneous (SCF), peri‐gonadal (PGF), peri‐renal (PRF) localisation were separated, weighed and flash frozen. Blood was collected and plasma was flash frozen for future measurement of glucose, insulin and triglycerides. Gene expression for glucose transporters GLUT1 and GLUT4 were assessed by real‐time PCR.Results(P)RR‐B administration lowered weight gain in the HF/HC fed mice, whereas food intake didn't differ between the groups. Increases produced by the HF/HC in circulating glucose, insulin and triglycerides were significantly decreased. Insulin sensitivity measured by glucose‐insulin ratio was also improved by the (P)RR‐B. Altered GLUT1 and GLUT4 mRNA expression in mice on a HF/HC was improved by the (P)RR‐B, in line with the circulating metabolites data.Conclusion(P)RR‐B administration seems to improve glucose homeostasis, insulin sensitivity and GLUTs gene expression profile in mice fed a HF/HC.Research supported by CIHR, HFSC, CDA and Merck Frosst Canada
Building better animal models of disease through integration of multiple disease phenotypes
Authors: Dwinell Melinda Ryan , Nigam Rajni , Liu Weisong , Smith Jennifer R , De Pons Jeff , Shimoyama Mary 
Published: June 2021

Animal models of disease are used to test hypotheses of disease mechanisms as well as for therapeutic development. Effective animal models must sufficiently reflect human disease and should be classified carefully as disease models. Integration of genomic and phenotypic data provides the foundation to study the genetic and environmental influences on the risk, onset and progression of disease. PhenoMiner, at the Rat Genome Database (http://rgd.mcw.edu), is an interactive data‐mining tool to compare clinical measurements used as indices of disease across multiple rat strains, measured under a variety of conditions. Data are integrated through the use of four ontologies (rat strain, clinical measurement, measurement method, experimental conditions). PhenoMiner provides a solid foundation to distinguish phenotypes by strains and strains by phenotypes. Currently more than 42,000 records can be viewed graphically or in tabular format, allowing easy comparison across strains. Graphical summaries (bar charts and heat maps) provide a rapid method to identify strains with abnormal phenotypes or groups of phenotypes that correspond to disease traits. In addition to providing phenotype driven model identification, PhenoMiner generates standard ranges for clinical measurements under control conditions to be used to identify strains with potential disease traits.Supported by R01HL094271 and R01HL064541.