bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rab11A and Rab11B distinctly regulate protease‐activated receptor‐1 recycling and constitutive degradation (802.10)
Authors: Coronel Luisa , Grimsey Neil , Canto Isabel , Stephens Bryan , Phan Cattien , Handel Tracy , Trejo JoAnn 
Published: June 2021

Protease‐activated receptor‐1 (PAR1) is a G protein‐coupled receptor (GPCR) expressed throughout the vasculature. Thrombin irreversibly activates PAR1, therefore receptor signaling and expression is tightly regulated. Upon activation, PAR1 is rapidly internalized and sorted to lysosomes for degradation, a process critical for signal termination. Whereas naïve PAR1 is constitutively internalized, generating an intracellular pool of PAR1 that recycles and repopulates the cell surface with uncleaved PAR1 after thrombin stimulation, a process critical for rapid recovery of thrombin signaling. The mechanisms that mediate recycling of naive PAR1 are not known. Using a siRNA library screen of 140 membrane trafficking proteins, we identified Rab11B as a key regulator of PAR1 trafficking. We confirmed that siRNA depletion of Rab11B, and not Rab11A reduced PAR1 surface expression, but did not affect expression of the thromboxane GPCRs, TPα and TPβ. Rab11B knockdown had no affect on PAR1 constitutive internalization, but blocked PAR1 recycling resulting in loss of surface expression and enhanced receptor degradation that was blocked by lysosomal protease inhibitors. In contrast to Rab11B, knockdown of Rab11A by siRNA disrupted PAR1 sorting from endosomes to lysosomes leading to accumulation, without affecting surface expression, suggesting that Rab11A and Rab11B distinctly regulate PAR1 trafficking. The mechanisms by which Rab11A and Rab11B distinctly regulate intracellular trafficking of PAR1 are not known and currently being investigated. Given that elevated PAR1 expression contributes to malignancy of certain cancers, understanding the mechanisms that control PAR1 surface expression could aid in the development of therapeutic approaches.Grant Funding Source: Supported by NIH R01 GM090689, Western States Affiliate AHA Postdoctoral Fellowship and the NIGMS In
Imaging kinase: connexin43 interactions using phosphorylation activity biosensors and phospho‐specific antibodies (802.2)
Authors: Cone Angela , Cavin Gabriel , Ambrosi Cinzia , Hakozaki Hiroyuki , Newton Alexandra , Sosinsky Gina 
Published: June 2021

Connexin43 (Cx43) is a ubiquitous gap junction protein with widespread tissue expression, serving important functions in cell homeostasis and signaling. The C‐terminus of Cx43 contains multiple phosphorylation sites and protein binding domains that are involved in regulation of connexin trafficking and channel gating. Several kinases phosphorylate Cx43 (Protein Kinase A, Protein Kinase C, Akt (also known as Protein Kinase B), Src, CK1, MAPK) at specific serines or tyrosines. We used kinase biosensors,for dynamic readouts of function, and combined it with image analysis to obtain spatial information. We fused a genetically encoded fluorescence resonance energy transfer‐based reporter for PKC delta activity, delta C kinase activity reporter (dCKAR) to the C‐terminus of Cx43 (Cx43‐dCKAR) and expressed it in Cos7 cells. Similarly, we genetically appended an activity reporter for Protein Kinase A (AKAR‐CR) to Cx43 and expressed it in Cos7 cells. Time lapse imaging of stimulated PKC resulted in phosphorylation of Ser368 Cx43‐dCKAR channels and discrete domains in the center of the gap junction, followed by internalization and degradation. Phosphorylated Cx43‐AKAR‐CR was predominantly found in the Golgi apparatus and to a lesser extent around the plaque edges. Thus, the spatio‐temporal localizations of PKC and PKA phosphorylated Cx43 represent degradative and synthetic pathways, respectively.Grant Funding Source: Supported by NIH grants GM072881, GM043154, GM103412
Scaffolding of atypical protein kinase C to p62 is disrupted by the pseudosubstrate peptide ZIP (802.4)
Authors: Tsai Li‐Chun , Xie Lei , Xie Li , Bourne Philip , Newton Alexandra 
Published: June 2021

Atypical protein kinase C (aPKC) enzymes, aPKCλ and aPKCζ, play key roles in metabolic signaling. A myristoylated peptide based on the autoinhibitory pseudosubstrate fragment of PKCζ, zeta inhibitory peptide (ZIP), has been extensively used to inhibit aPKC activity, however we have recently shown that ZIP does not inhibit the catalytic activity of aPKC isozymes in cells (Wu‐Zhang et al., (2012) JBC 287:12879‐85). Here, we identify a novel interaction between the pseudosubstrate segment of PKCλ and the protein scaffold p62, a known binding partner, that is disrupted by ZIP in cells. First, we used protein‐protein interaction network analysis, structural modeling, and protein‐protein docking to predict that ZIP, a basic peptide, binds an acidic surface on the PB1 domain of p62. This predicted interaction is additional to the known interaction between the PB1 domain of aPKC, which immediately precedes the pseudosubstrate segment, and p62. Second, we used fluorescence resonance energy transfer (FRET) experiments to show that ZIP disrupts the interaction of CFP‐tagged p62 and YFP‐tagged PKCλ in cells via a mechanism that depends on an intact pseudosubstrate segment. In contrast, auranofin, a known disrupter of the PB1:PB1 domain interaction of aPKC and p62, disrupts the interaction of wild‐type PKCλ, a construct lacking the pseudosubstrate, but not a construct in which the PB1 domain is deleted. Our data are consistent with ZIP altering cellular function by displacing binding partners from the acidic face of the PB1 domain of p62, a region which we identify as an interaction surface for the pseudosubstrate segment of aPKCs.Grant Funding Source: This work was supported by 5T32DK007494 and P01 DK054441.
Trend in caffeine intake and food and beverage sources of caffeine in U.S. children over the last decade: analysis of What We Eat In America/National Health and Nutrition Examination Survey (NHANES) 2001‐2010 (806.17)
Authors: Fulgoni Victor , Schorin Marilyn 
Published: June 2021

WWEIA data and the food groupings within were used to assess caffeine intake and food and beverages sources of caffeine in the diets of children and adolescents 6‐18 years of age (N=13,131) over the last decade. Regression analyses were conducted to determine a p‐for‐trend across the NHANES data releases (2001‐2002, 2003‐2004, 2005‐2006, 2007‐2008, and 2009‐2010). For the total population, total caffeine intake remained stable across the five NHANES releases (beta coefficient: ‐0.5 mg/release; p= 0.6191). Major contributors of caffeine (mean ± SE, mg/d) in the diet across the decade were regular soda (42±2); tea (10±1), coffee (6±1), diet soda (2±0.3), and energy drinks (1±0.3). There was a significant p‐for‐trend of ‐3 mg/release for regular sodas (p<0.0001) and 1.1 mg/release for coffee (p<0.01). Energy drinks increased 0.3 mg/release (p<0.05), but their contribution was only 1 mg/d. Among caffeine consumers (N=9,400), total caffeine intake remained stable across the five NHANES releases (beta coefficient: ‐0.2 mg/release; p= 0.8581). Also among caffeine consumers, caffeine contributed by each beverage category evaluated remained stable across the decade. For the total population, total caffeine intake has remained stable among children over the decade with a decrease in contribution from soft drinks and a slight increase from coffee.Grant Funding Source: American Beverage Association
Nutritional implications of marijuana use in an HIV infected population in the Miami Adult Studies on HIV cohort in Miami (810.7)
Authors: Kienzle Cinda , Campa Adriana , Kurtz Joy , Li Yinghui , Greer Pedro , Martinez Sabrina , Stewart Tiffanie , Boyer Jacqueline , Baum Marianna 
Published: June 2021

OBJECTIVE: To determine the relationship of nutritional status and MJ use in HIV+ population .METHODS: Dietary intake, food insecurity, plasma levels of nutrients, anthropometrics, CD4 count, viral load and ART use were determined with consent in 201 HIV+ participants. Multivariate and logistic regressions were used.RESULTS: Mean age was 45.2 years, mostly males (64%) and Blacks (72.5%); 27% used MJ. MJ users were younger (mean age 42 vs. 46, P<0.001) and used alcohol and tobacco more frequently than non‐users (94% vs. 87%, P<0.001). MJ use was associated with better ART adherence (β=0.99, P=0.043), higher CD4 cell count (β=112.87, P=0.03) and lower homocysteine (β=‐1.05, P=0.051). MJ users trended toward increased BMI (β=1.47, P=0.08) and %fat mass (β=3.02, P=0.08). In contrast, MJ users were more food insecure (52% vs. 33%, P=0.015) and had lower % body cell mass (β=‐3.3, P=0.018) and serum iron (β=‐13.16, P=0.018). Caloric intake did not differ between MJ users and non‐users.CONCLUSION: MJ use is associated with some positive effects on HIV disease progression, but the effect on body composition seems to be from fat accumulation. Studies are needed to determine the effects of MJ on health in HIV infection.Grant Funding Source: NIDA and NIAAA
Body image and weight control in youth: 9‐year international trends from 24 countries (811.5)
Authors: Quick Virginia , Nansel Tonja , Liu Danping , Lipsky Leah , Due Pernille , Iannotti Ronald 
Published: June 2021

This study’s purpose was to examine 9‐year trends in misperceptions of body size and dieting for weight loss among adolescents from 24 countries, and explore the influence of country‐level overweight prevalence. Body size perception and dieting for weight loss were assessed in the Health Behaviour in School‐aged Children survey conducted in 24 countries cross‐sectionally at 3 time points (2001/02, 2005/06, 2009/10). Logistic regression models examined change over time in over‐ and under‐estimation of body size and dieting for weight loss. Compared to Time 1, overweight adolescents had greater odds of body size underestimation at Time 3 (OR=1.68 for girls, OR=1.10 for boys), while non‐overweight adolescents had lower odds of body size overestimation at Time 3 (OR=0.87 for girls, OR=0.89 for boys). Controlling for country‐level overweight prevalence attenuated these relationships. Compared to Time 1, overweight and non‐overweight boys were 10% more likely to diet at Time 3, while overweight and non‐overweight girls were 19% and 16%, respectively, less likely to diet at Time 3. Controlling for country‐level overweight prevalence did not impact trends in dieting for weight loss. Findings suggest body size perceptions among adolescents may have changed over time concurrent with shifts in country‐level body weight.Grant Funding Source: Supported by Maternal and Child Health Bureau of the Health Resources and Services Administration
Fruit juice, fruit drinks and atole are associated with obesity and lipid profile in school‐aged children from rural México (811.8)
Authors: Garcia Olga , Zavala Gerardo , Caamaño María , Ronquillo Dolores , Martínez Guadalupe , Camacho Mariela , Rosado Jorge 
Published: June 2021

Beverages such as fruit drinks (natural and artificially flavored), soft drinks and 100% fruit juices are regularly consumed by children as part of their diet. 198 children (8 ± 1.5 years) participated in a cross‐sectional study to evaluate the association of beverage intake with obesity, lipid profile and glucose concentration. Body fat (DXA), height, weight and waist circumference were measured in all children. Diet was evaluated with a food frequency questionnaire and three 24hr recalls. A fasting blood sample was taken to analyze lipid profile and glucose. 17% of the daily energy intake of these children came from beverages. Linear regressions adjusted for socio‐demographic variables showed some significant associations (p<0.05). High intake of 100% fruit juices was associated with high body and abdominal fat, waist‐to‐height ratio, and high triglyceride concentration. Fruit drinks were positively associated with higher BMI z‐score and triglycerides. Intake of atole (maize based beverage with added sugar) was associated with high triglycerides levels. Milk intake was associated with lower LDL concentration. Soda intake was not associated with any of the variables studied. In conclusion, high intake of 100% fruit juices, fruit drinks and atole in school‐aged children from rural México, are associated with obesity and alterations in lipids profile.Grant Funding Source: Supported by CONACYT
Evaluation of protein‐phytochemical interactions in a hypoallergenic edible peanut matrix (813.10)
Authors: Plundrich Nathalie , White Brittney , Davis Jack , Lila Mary Ann 
Published: June 2021

Allergenic peanut proteins (PP) are stable to digestion, remain immunoreactive and trigger IgE‐mediated allergic reactions in PP sensitive individuals. In this study, a hypoallergenic peanut protein‐phytochemical matrix (PPPM), produced by complexing cranberry juice (CB) with light roast peanut flour, was evaluated to determine the nature of protein‐phytochemical interactions and the stability of the binding post‐ingestion. PPPM or unmodified peanut flour were subjected to a simulated gastric fluid digestion. Aliquots of the enzymatic mixture were taken at different time points and inhibited with either sodium bicarbonate (pH ~7‐8) or heat (pH ~2) to stop the digestive process. Digested PP fragments and undigested PP were examined on SDS‐PAGE and screened for their peanut‐specific IgE binding capacity using Western blotting, respectively. SDS‐PAGE revealed that PP were more soluble at a low pH than at a basic pH. PP treated with CB had reduced solubility at both acidic and basic pH, compared to untreated PP. However digestion of both PPPM and unmodified peanut flour resulted in formation of highly soluble small molecular weight PP fragments at either pH. Examination of the Western blots revealed that digested PP fragments or undigested PP from PPPM exhibited altered IgE binding, compared to proteins from unmodified peanut flour. Modification of PP with CB phytochemicals led to the formation of insoluble complexes with altered IgE binding to the protein epitopes, suggesting altered PP digestion and IgE binding capacity.Grant Funding Source: Supported by Everett W. Byrd Endowment and NC State appropriations for NCSU at Kannapolis
A 14‐day in‐house dietary modification of a ‘Western’ diet to an ‘African’ diet changes the microbiota, its metabolome, and biomarkers of colon cancer risk (825.5)
Authors: Vipperla Kishore , Ou Junhai , Wahl Elaine , Ruder Elizabeth , O'Keefe Stephen 
Published: June 2021

Methods Twenty age and sex matched middle aged healthy African Americans were studied on their usual diet and then again after dietary modification to a low meat and fat, high fiber ‘traditional African’ diet, prepared and delivered individually in‐house under close supervision for 2 weeks. Phutu‐ corn meal, Samp‐ dried corn kernels, sugar beans, fruits and vegetables provided 30‐35 Kcal/Kg of energy, 14% protein, 17% fat, 72% carbohydrate, 40 g dietary fiber, and 32 grams resistant starch. The diet was well tolerated overall and their body weights remained stable. Pre‐ and post‐dietary modification fecal and colonic evacuate samples were collected to measure the microbiota and their metabolites and colonoscopies were performed to assess mucosal health and to obtain biopsies for the measurement of inflammatory and proliferative biomarkers (Ki67‐ identified by immunohistochemical staining).Results Following dietary change, significant increases were measured in the expression of functional microbial genes encoding butyrate (BcoA gene, 1.1E+10±1.8E+10 vs. 2.5E+10±9.3E+09 copies/g feces, p= 0.04) and methane synthesis (mcrA gene, 6.5E+5±1.2E+5 vs. 1.6E+6±3.9E+5 copies/g feces, p= 0.01); fecal short chain fatty acids, e.g. butyrate concentrations (3.56±0.96 vs. 8.61± 2.10 µmol/gm feces, p=0.034); expired breath methane levels (12.8±3 vs. 20.3±8.3 ppm, p=0.02); Finally, mucosal Ki67 expression was significantly reduced (p<0.0001).Conclusion In this study, short term dietary change in high colon cancer risk African Americans to a low meat and fat, and high fiber “African” diet promoted saccharolytic fermentation and the production of anti‐neoplastic metabolites such as butyrate in association with a reduction in biomarkers of colon cancer risk. These results support the recommendations for reducing meat and fat and increasing fiber to suppress colon cancer in high‐risk populations.Grant Funding Source: Supported by National Institutes of Health, R01 CA135379 (O’Keefe) and UL1 RR024153 and UL1TR000005
Cellular pyridoxal phosphate restriction in cultured HepG2 cells decreases formation and flux of metabolites in the transsulfuration pathway including production of hydrogen sulfide biomarkers (827.6)
Authors: DeRatt Barbara , Ralat Maria , Kabil Omer , Banerjee Ruma , Gregory Jesse 
Published: June 2021

Cystathionine beta‐synthase (CBS) and cystathionine gamma‐lyase (CSE) are pyridoxal phosphate‐dependent transsulfuration enzymes responsible for the majority of endogenously produced hydrogen sulfide (H2S) and concurrently formed biomarkers, lanthionine (lan) and homolanthionine (hlan). H2S modulates physiological functions in the cardiovascular and central nervous systems. We investigated the effects of restricted vitamin B6 supply on cultured HepG2 cell metabolism by analyzing transsulfuration pathway kinetics. Cell culture media contained pyridoxal (PL) concentrations at severely deficient (10 nM), marginal (50 nM), adequate (200 nM) and supraphysiological (2000 nM) levels. Metabolic flux studies using [U‐13C5] L‐methionine and (3,3 D2) L‐cysteine indicated lower fractional synthesis rates (FSR) and steady state concentrations of lan and hlan at 10 and 50 nM PL (P<0.027). The FSR of cystathionine was higher in cells cultured in 50 nM PL than other PL concentrations (P<0.001), consistent with the greater sensitivity of CSE than CBS to B6 deficiency. In‐vitro H2S production capacity in assays with cell lysates and these results confirmed that H2S production decreased in B6 deficiency. These findings support a hypothesis that impaired H2S production may be involved in the association between vitamin B6 inadequacy and cardiovascular disease.Grant Funding Source: Supported by NIH Grant DK072398
Effect of vitamin D on ROS, ICAM‐1, and monocyte adhesion in human umbilical vein endothelial cells treated with high glucose and acetoacetate (828.10)
Authors: Kanikarla Marie Preeti , Jain Sushil 
Published: June 2021

Monocyte adhesion to the endothelium plays a central role in the progression of endothelial dysfunction and cardiovascular disease (CVD). This study examines whether VD supplementation is beneficial in preventing endothelial dysfunction in diabetes. HUVEC were treated with 1, 25‐(OH)2‐D3 (0‐25nM, 24h) and later exposed to the ketone body acetoacetate (AA, 4mM) or high glucose (HG, 25mM) for 24h. There was an increase in ROS, ICAM‐1 expression, and monocyte adhesion in HUVEC treated with AA or HG. VD (25nM) supplementation significantly lowered ROS (41%, p<0.05), ICAM‐1 (73%, p=0.05) and p‐NFκB expression (50%, p<0.05), and IL‐8 (16%, p=0.05) and MCP‐1 (41.2%, p<0.001) secretion along with monocyte adhesion (37.5%, p=0.05) in AA treated HUVEC. VD had a similar beneficial effect on HG treated HUVEC. Interestingly, we also saw an upregulation in GSH and GCLC with VD. We hypothesize that the beneficial effect of VD is mediated by the upregulation of GCLC and GSH, a potential antioxidant. To explore this idea, we knocked down GCLC using siRNA. Results show that beneficial effect of VD was abolished on ketone‐ or HG‐induced ICAM‐1 in GCLC knockdown cells. This suggests that VD mediates its beneficial effects via the upregulation of GCLC. This study provides the biochemical mechanism through which VD can lower ROS, ICAM‐1 expression, monocyte‐endothelial adhesion, and thereby lower the risk of CVD in diabetes.Grant Funding Source: Supported by NIH and Malcolm Feist Chair in Diabetes
Grape pomace bioactivity from Brazilian wine industry: toxicity and antioxidant properties in human hepatocarcinoma cells (830.31)
Authors: El Bacha Tatiana , Sales Nathalia , Minuzzo Daniela , Cruz Ana Paula , Cabral Lourdes , Torres Alexandre 
Published: June 2021

Wine industry produces large quantities of by‐products rich in bioactive compounds from grapes, which could be utilized for nutraceutical purposes. We aim to evaluate antioxidant activity (AC) and bioactivity of grape pomace extract from Brazilian wine industry in human hepatocarcinoma cells (HepG2). The content of bioactive compounds and AC of extracts were determined by ORAC, TEAC (ABTS•‐) and TRAP assays. Bioactivity was assessed on HepG2 and on normal human fibroblasts (BEAS). Cellular viability was accessed by MTT reduction and cellular antioxidant capacity (CAC) by dichlorofluorescein oxidation, under short‐, medium‐, and long‐term incubation periods. Total phenolics, flavonoids and anthocyanins of extracts where, respectively, (mean±SD) 473.8±52.4 mg GAE/100g, 147.5±2.77 mg CE/100g and 105.5±7.27 mg cyanidin3‐glycoside/100g. Extract AC was (mean±SD; µmol TE/g) 27.1±0.33, 9.33±6.74, 122.2±2.40. HepG2 viability reduced in a time‐ and dose‐dependent manner. Short‐term incubation had no effect whereas medium‐ and long‐term incubation induced, respectively, a 37% and a 75% reduction in viability. BEAS viability was not affected by the extract, regardless of time and concentration used. Interestingly, short‐term incubation promoted a dose‐response increase in HepG2 CAC, suggesting that decreased oxidative stress signals to decrease HepG2 proliferation. Taken together, wine industry by‐products present important antioxidant activity and potential selective anticancer and antiproliferative effects on HepG2 cells.Grant Funding Source: Supported by FAPERJ, CNPq, CAPES, EMBRAPA (Brazil)
Dematin and its interaction with platelet cytoskeleton (833.8)
Authors: Azim Anser , Murungi Terry , Giragandi Abdelrahim 
Published: June 2021

Background: Human platelets have spectrin based cytoskeleton that supports the plasma membrane of platelets and plays an important role in its activated and resting stages. The platelet cytoskeleton is also composed of peripheral microtubules. Human platelets have many actin binding proteins like spectrin, α□actinin, vinculin, talin, trpomyosin, adducin, dematin, myosin etc. In platelets monomeric actin is rapidly polymerized into F‐actin upon stimulation with the formation of surface filopodia and lamellipodia and acquires dendritic shape. Myosin forms stress like fibers with actin molecules. Dematin plays a role in activated platelets in the regulation of cytoskeleton. In this study we hypothesize the role of dematin, an ATP binding protein, in the regulation of cytoskeleton and its interaction with actin binding proteins.Method: We isolated human platelets from anti‐coagulated blood as described earlier (Azim AC et al Blood. 2000, 95(3):959‐64.). Platelets were used after incubating them at 37 0C for half an hour before the experiment.Results: The present study serially analyzed dematin’s coimmunoprecipitation with platelet myosin. And we believe that this interaction serves as a possible mechanism of ATP loading on the myosin molecule that subsequently plays a role in spreading and shape change.Discussion: In an attempt to understand the physiological role of dematin in hemostasis its worth knowing the role of this actin bundling protein, dematin, in human platelets. We show for the first time that platelet myosin interacts with dematin. Further studies are needed to know the precise role of this interaction in regulating actin cytoskeleton, shape change and platelet expansion.