bluuebunny/crossref_metadata_2025 huggingface dataset:
The Two‐pore channel 2 (TPC2) mediates autophagy in skeletal muscles
Authors: Lin pei‐hui , Komazaki Shinji , Zhu hua , Park ki ho , Pan zui , Duann pu , Ma jianjie
Published: June 2021

Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent Ca2+‐mobilizing messenger that in many cells releases Ca2+ from the endolysosomal system. Recent studies have shown that NAADP‐induced Ca2+ mobilization is mediated by the two‐pore channels (TPCs). The objective of this study is to test whether TPC2 is involved in autophagy in skeletal muscle.

Methods

Wild type or TPC2 null animals were subjected to autophagy induction to compare the autophagy fluxes in the animals. In some experiments, isolated neonatal myoblasts were used. Detection methods include LC3 western blot and electron micrographs.

Summary of results and conclusions: Our data provide evidence that TPC2 is involved in regulating autophagy flux in skeletal muscle. TPC2 mutant demonstrated defective autophagy to maintain tissue homeostasis.

Summary of funding: There is no NIH funding to support this research project yet.
BUMPT cells as a model system for studying olfactory receptor signaling in renal epithelial cells.
Authors: Natarajan Niranjana , Pluznick Jennifer L
Published: June 2021

Olfactory receptors (ORs), a class of chemosensory G‐protein coupled receptors, are expressed in non‐sensory tissues (including kidney) where they serve to detect environmental cues. Renal ORs are expressed in cell types that lack components of the canonical OR signaling, Golfactory protein (Golf) and adenylate cyclase (AC3), indicating that they must signal via an unknown alternate signaling pathway involving a native G protein. We find that BUMPT, a mouse proximal tubule‐derived cell line, expresses 5 of the 9 known renal ORs including OR 691 (Olfr691; RT‐PCR). We also find that BUMPT cells, like native proximal tubule cells, do not express Golf or AC3 (RT‐PCR). Therefore, we used BUMPT to investigate the non‐canonical signaling mechanism of renal ORs. Using an unbiased degenerate PCR primer screen for Gα subunits, we identified Gα11, Gq and GαS in BUMPT cells (consistent with Gα subunits in the PT proteome). Gα11, Gq and GαS were subsequently confirmed to be expressed in both BUMPT and mouse kidney using gene‐specific primers. We have confirmed Gα11 and Gq protein expression in BUMPT cells and mouse kidney using an antibody that recognizes both; efforts to confirm the expression of GαS are in progress. We plan to test for physical association of Gα11, Gq and/or GαS to Olfr691, and investigate second messengers and downstream targets in BUMPT cells to elucidate the mechanism of renal OR signaling. Funding: NIDDK
Orthogonal In Situ Quantification of Multiple Cellular Lipids
Authors: Afsari Hamid Samareh , Kurilova Svetlana , Cho Wonhwa
Published: June 2021

Lipids are dynamic molecules whose local concentrations can vary to a large degree under physiological conditions. In order to fully understand diverse lipid‐mediated cellular processes, it is therefore necessary to have a sensitive analytic tool for monitoring their local fluctuations. Recently we developed a new chemical method to quantify cellular lipids in situ using a molecular sensor made of an engineered lipid binding protein that is labeled with an environmentally sensitive fluorophore. Here, we report a new and advanced method employing two orthogonal fluorphores that allows simultaneous in situ quantification of two membrane lipids in live cells. This new methodology serves as a powerful tool for systems‐level investigation of lipid‐mediated cell signaling and regulation.
The Involvement of SWI/SNF chromatin remodeling enzymes in Transcriptional Regulation of Pathological Cardiac Hypertrophy
Authors: Mehrotra Aanchal , de la Serna Ivana L
Published: June 2021

Pathological cardiac hypertrophy is characterized by transcriptional reprogramming of gene expression which involves certain chromatin modulators. Little is known about the chromatin modifications involved in hypertrophy. SWI/SNF chromatin remodeling enzymes are epigenetic modulators playing pertinent roles in gene regulation. To understand the involvement of various subunits in the SWI/SNF complex and to study the chromatin modifications occurring in hypertension induced cardiac hypertrophy, we performed expression analysis as well as chromatin immunoprecipitaions (CHIPs) on Dahl salt sensitive and resistant rat hearts. The relative protein expression revealed a significant increase of Brg1, Baf 60c and Baf 180 in sensitive rat hearts. Moreover CHIPs demonstrated an increased enrichment of these subunits on promoters of ANP and BNP (hypertrophic markers) in sensitive models. Also there was an increased relative enrichment of acetylated H4, H3K4 trimethylation (active transcription marks) and decreased enrichment of H3k9 trimethylation, H3k27 trimethylation (negative transcription marks) on ANP and BNP promoters in sensitive rats. Our studies for the first time revealed the importance of SWI/SNF complex in promoting cardiac hypertrophy in Dahl salt sensitive and resistant models and suggest what we believe to be a novel mechanism for reprogramming of gene expression involved in hypertrophy.
Epigenetic regulation of Cytochrome P450 (CYP1A1) by polycyclic aromatic hydrocarbons
Authors: del Rosario Olguin Sitlali , Camacho Rafael , Espinosa Jesus Javier
Published: June 2021

Gene regulation involves transcription factors, responsive elements, enhancers, insulators, DNA methylation, histone methylation, acetylation, and deacetylation, among others. In an organism, such components might depend on external factors such as environmental pollution. CYP1A1 gene is inducible by polycyclic aromatic hydrocarbon (PAH) through the aromatic hydrocarbon receptor (AhR). However, exposure to PAHs may affect proteins involved in epigenetic regulation, as well as, CYP1A1 epigenetic regulation. In order to probe this, rat hepatocytes clone‐9 cell line (C9) was exposed to benzo[a]pyrene (BaP), benzofluorene (BF), 2’‐7’‐dimethylbenzantracene (DMBA), 3‐methylcholanthrene (3MC) and DNMTs inhibitor 5’‐Aza‐2’‐deoxycitidine (5AzadC).

CYP1A1 gene expression was determined by qPCR and DNMT3a protein level by western blot, showing in both cases an increased with all treatments. However, this effect is exacerbated with a PAH‐AzadC treatment, especially with DMBA. Additionally, treatments also increase DNMT3a protein level in C9 culture. In respect to the positive modulation of CYP1A1 gene expression, we suggest that PAHs do not affect CYP1A1 gene expression only through AhR, but it is also affected by its epigenetic regulation.
Parents education and children nutritional status aged 2 to 5 in Zambia (1014.8)
Authors: Alalaq Hassan , Katuli Sozian , Beeson Lawrence , Ormsby Gail , Cordero‐MacIntyre Zaida
Published: June 2021

B: The aim of this study is to evaluate the independent association of parental education on the nutritional status of the children using anthropometric measurements and body composition tools. M: The WHO standardized age and sex‐specific growth reference was used to calculate height for age Z‐scores (HAZ), weight for age Z‐scores (WAZ) and weight for height Z‐scores (WHZ). Percent body fat was calculated using the equation developed by Lohman for skinfold. Logistic regression was used to evaluate the relationships of parent’s education and the prevalence for stunting controlled for family size.R: 77 children were assessed (M=32, F=45) of whom 45(58.4%) were stunted expressed by HAZ <‐2 (Mean Z score ‐2.33±1.24) and 18 (23.3%) had acute malnutrition expressed by WAZ <‐2 (Mean Z score ‐1.25±1.12).

About 41(55.4%) children were born by mothers who had no education and 20 (29.8%) whose fathers had no education. The logistic regression showed that parent’s education was not significantly associated with children nutritional status (p‐value 0.3037).

C: Our results indicate that more than half of the children are stunted indicating chronic malnutrition. This may be a reflection of chronic food shortage or poverty which may affect food intake by the children. Parent’s education seems to be insignificant strategy to improving children’s nutrition in Zambia. Further studies are needed to assess other health problems in the area.

Grant Funding Source: The Center for Health Research, Loma Linda University, and in participation with ADRA Zambia Child A
Yogurt consumption is associated with lower body weight status and improved nutrient intakes in adult women (1018.11)
Authors: Gugger Carolyn , Joshi Nandan , Albertson Ann
Published: June 2021

This study aimed to determine estimates of yogurt consumption, nutrient intakes and body mass index (BMI) in U.S. women using the Bell Institute of Health and Nutrition Dietary Intake Study. Fourteen‐day (14‐d) diet records from a nationally representative sample of women 20+ years (n=2883) were collected over a 2‐year period. Yogurt consumption was determined over the 14‐d period, and participants were classified into intake categories for descriptive purposes: 0 servings, 1‐2 servings, 蠅3 servings. BMI was calculated from self‐reported height and weight. Data were analyzed using regression models adjusting for age, energy, calcium, vitamin D and fiber intake. Over 60% of women reported consuming no yogurt over the 14‐d period, with 19.3% consuming 蠅3 servings. Non‐yogurt eaters had significant lower intakes of protein, calcium, potassium and vitamin D compared to women who consumed yogurt (p<0.001). Furthermore, women who consumed yogurt were more likely to meet daily recommendations for calcium intake (p<0.001). Women who consumed at least one serving of yogurt had a significantly lower BMI compared to those consuming no yogurt over the 14‐d period (p=0.001). In addition, more frequent yogurt consumption (蠅3 servings/14‐d) was associated with a significantly lower incidence of overweight/obesity (p<0.05). These data suggest that regular consumption of yogurt may play a role in helping adult women maintain a healthier body weight and can contribute to more favorable nutrient intakes.
Evaluating the effectiveness of a nutrition‐sensitive agriculture intervention in Western Kenya: design of the Mama SASHA cohort study of vitamin A (1019.2)
Authors: Webb Girard Amy , Grant Frederick , Okuku Haile , Akelo Victor , Wanjala Rose , Levin Carol , Cole Donald , Low Jan
Published: June 2021

The 2013 Lancet Maternal and Child Nutrition series identified evaluations of nutrition‐sensitive agricultural interventions as a research priority. The Mama SASHA study in Western Kenya links delivery of vitamin A (VA) rich orange‐flesh sweet potato (OFSP) to antenatal care to improve VA and nutritional status of pregnant and lactating women and their children. The evaluation strategy includes a nested longitudinal study following women and their infants from pregnancy to 9 months postpartum. VA status is assessed using infection‐adjusted plasma retinol binding protein and breastmilk retinol (postpartum). Maternal and child infection, iron, and anemia status, anthropometry, diet, health services uptake, and food security status are also measured. 505 eligible pregnant women, attending ANC at 4 control and 4 intervention facilities, were consented and enrolled. At enrollment women in control and intervention communities did not differ with respect to VA, iron, anemia or anthropometric status; food security or dietary diversity scores; demographic characteristics; awareness of vitamin A; or consumption of vitamin A rich foods in the past 7 days. Only 10 women consumed OFSP in the previous 7 days; all in intervention communities. The longitudinal study will contribute to rigorous evaluation of the OFSP intervention on maternal and child VA status and allow assessment of program impact pathways.

Grant Funding Source: Bill and Melinda Gates Foundation
Seasonality in women’s dietary intake: preliminary results from the Life in all Seasons study (1022.5)
Authors: Jahns Lisa , Johnson LuAnn
Published: June 2021

Objective: Seasonal variation in foods consumed may bias estimates of long‐term food and nutrient intake in observational studies. We present dietary results from the first cohort of subjects in a year‐long study of predictors of change in body composition.

Methods: Longitudinal design; 27 women ages 40‐60 y completed online diet recalls approximately every 10 days (N=972) using the Automated Self‐administered 24‐hour Recall system. Reported intakes were compared to Dietary Guidelines for Americans (DGA) recommendations and seasonal differences tested by mixed model analysis of variance.

Results: Mean energy intake was not significantly different from estimated requirements (P = 0.23). Compared to Dietary Reference Intakes, recommendations, intakes of fiber, vitamin E, potassium, and vitamin D were inadequate and did not differ by season. Over the year, women reported consuming 63 ± 3% of the DGA recommended number of servings for vegetables, fruits (73 ± 10%); whole grains (36 ± 4%), seafood (54 ± 9%) and dairy (63 ± 5%). Grain intake was significantly higher in the fall compared to spring and summer (P<0.05) and intake of “other” vegetables was higher in summer than winter (P<0.05).

Conclusions: Nutrient intake did not vary seasonally in this sample of women. Intakes of key nutrients and nutrient‐rich food groups of public health significance were suboptimal.

Grant Funding Source: Supported by USDA‐ARS 5450‐51000‐049‐00D
Gender effects of supplemental amino acids on non‐alcoholic fatty liver disease and chronic inflammation (1025.14)
Authors: Belalcazar L. Maria , Spratt Heidi , Green Melanie , Volpi Elena , Hurren Nicholas , Wolfe Robert , Børsheim Elisabet
Published: June 2021

Background: Non‐alcoholic liver disease increases with age. There is scant information on gender differences in relation to its behavior, health consequences and treatment response.

Aims: 1). To examine gender effect on changes (Δ) in liver fat, body fat, and high‐sensitivity C‐reactive protein (hs‐CRP), a marker of subclinical inflammation and increased cardiovascular risk, in response to supplemental essential amino acids (sEAA); 2) To correlate Δ in liver and body fat with hs‐CRP Δ.

Methods: 12 subjects (7 women) with impaired glucose tolerance, age 67 years, ingested 22 g of sEAA daily for 8 weeks, after a placebo run‐in. Participants did not change diet or physical activity. Liver fat (nuclear magnetic spectroscopy; n=7), body fat (DEXA) and hs‐CRP levels (ELISA) were measured.

Results: Liver fat (0.403 to 0.209,intrahepatic triglyceride/Intralipid standard) and hs‐CRP (2.28 to 1.55 mg/L), but not body fat, decreased at 8 weeks from baseline. Analysis of variance showed a gender effect (p=0.047) for Δ in liver fat and hs‐CRP. Liver fat Δ, but not body fat Δ, was associated with hs‐CRP Δ in women (Spearman coefficients 0.63, p=0.067 overall; 0.9, p=0.037 in women; p=0.2 in men).

Conclusion: There may be significant gender differences on the effects of sEAA on hepatic steatosis and hs‐CRP levels.

Grant Funding Source: AG033761 , D34HP18956, UL1TR000071‐05, AG024832
Health status of a low socioeconomic elderly population that participates in a community senior meal program (1026.4)
Authors: Tsunoda Heather , Min Jong Won , Piwowarski Maureen , Hong Mee Young
Published: June 2021

The elderly population is one of the most vulnerable populations in terms of health. One of the most related factors to poor health in the elderly is low socioeconomic status. Those with the poorest health status are individuals with high poverty rates. Food insecurity, limited access to medical care, and decreased physical activity contribute to the low health status in the elderly. Those with low socioeconomic status put this vulnerable population at even higher risk of being either over or under nourished. The objective of this study was to conduct an initial assessment of health in a lower socioeconomic elderly population. 93 elderly individuals who participate in a San Diego senior meal program were assessed. Average body mass index (BMI) was 28 indicating overweight status and percent ideal body weight was 130%. 64.5% were considered clinically overweight or obese, and 3.5% underweight. Mean body fat was higher than normal ranges: 31% in male participants and 37% in female participants. 37.5% of participants reported <30 min daily exercise. Physical activity showed an inverse relationship with BMI value (P=0.07). Seniors reporting higher body fat was strongly correlated with high BMI (P<0.01). Data raises awareness of nutritional/health status of vulnerable seniors living in poverty. Further studies are needed to establish the role of food choice on nutritional status changes.
Effect of diet intervention on inflammation‐related gene expression in CD14+ circulating monocytes from metabolic syndrome patients (1037.10)
Authors: Martin Lorena , Deo Simmi , Sears Dorothy
Published: June 2021

Study objective: To examine the effects of a dietary intervention on circulating monocytes in patients with Metabolic Syndrome.

Methods: In this study, we analyzed pro‐inflammatory gene expression in CD14+ circulating monocytes from metabolic syndrome subjects before, during, and after two diet interventions. The study was a randomized, single‐blind, controlled trial of a 12‐week long, 1500 calorie diet. The active diet included low glycemic index bread products, EPA/DHA fish oil capsules (2.4g EPA+DHA/day), and delphinidin polyphenol capsules (300 mg, 10,000 ORAC units/day). The placebo diet included market variety bread products, corn oil capsules, and corn starch capsules. We hypothesized that the active dietary components would act synergistically to significantly improve the inflammatory gene expression profile of the subjects as compared to placebo diet. Baseline, midpoint, and endpoint mRNA expression profiles from monocyte RNA samples were assessed for 30 obese subjects.

Results: Post‐hoc analysis of real‐time quantitative polymerase chain reaction (RT‐qPCR) results to measure the expression levels of twelve inflammatory marker genes of interest revealed significant reduction in toll‐like receptor 2 (TLR2) F (1, 16) = 9.136, p < 0.01, CD11c, F (1, 16) = 12.484, p < 0.01 and tumor necrosis factor alpha (TNFα), F (1, 16) = p < 0.05 expression in circulating monocytes of active group as compared to the placebo group, p <0.05.

Conclusions: This study was part of a parent clinical trial showing that average fasting insulin and HOMA‐IR improved significantly in the active group but not the placebo group. This monocyte gene expression data will contribute to painting the picture of how the composition of a low calorie diet modulates inflammation and insulin sensitivity.

Grant Funding Source: Supported by P. Robert Majumder Charitable Foundation, UCSD CTRI (DDS), NIH (SKD), and Zone labs.
High rates of riboflavin deficiency in women of childbearing age in Cambodia and Canada (1041.12)
Authors: Whitfield Kyly , McCann Adrian , Karakochuk Crystal , Talukder Aminuzzaman , Ward Mary , McNulty Helene , McLean Judy , Green Tim
Published: June 2021

High rates of riboflavin deficiency in women of childbearing age in Cambodia and Canada Whitfield, KC1, McCann, A2, Karakochuk, C1, Talukder, A3, Ward, M2, McNulty, H2, McLean, J1, Green TJ1 (1) Department of Food, Nutrition, and Health, University of British Columbia, Canada (2) Northern Ireland Centre for Food and Health, University of Ulster, Northern Ireland (3) Helen Keller International, Phnom Penh, Cambodia As an integral component of the coenzymes flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN), riboflavin is essential for metabolic homeostasis. Limited evidence is available comparing riboflavin status among individuals from developed and developing countries.

We conducted two cross‐sectional studies with apparently healthy non‐pregnant, non‐lactating women of childbearing age (20‐45y), on a representative sample from urban Phnom Penh (n=146) and rural Prey Veng (n=156), Cambodia, and a convenience sample of urban women in Vancouver, Canada (n=51). As a functional biomarker of riboflavin status, erythrocyte glutathione reductase activity coefficient (EGRac) was measured.

Among Cambodian women, riboflavin deficiency (EGRac 蠅1.4) was prevalent (Prey Veng, 82%; Phnom Penh, 78%). Although a significantly higher proportion of Canadian women were riboflavin sufficient (EGRac 蠄1.3; p<0.001), 67% of Canadians had a suboptimal riboflavin status (EGRac >1.3).

The data suggests suboptimal riboflavin status and deficiency may be widespread in both developed and developing countries and merits further investigation.

Grant Funding Source: Supported by the University of British Columbia Vitamin Research Fund
Serum 25‐hydroxy vitamin D is associated with blood total cholesterol and diastolic blood pressure in a young multi‐ethnic group living in South Florida (1041.15)
Authors: Ajabshir Sahar , Exebio Joel , Zarini Gustavo , Nayer Ali , Vaccaro Joan , Shaban Lemia , Huffman Fatma
Published: June 2021

Objective: The aim of this study was to investigate the relationship between serum 25‐hydroxy vitamin D, total cholesterol and blood pressure (BP) among young multi‐ethnic participants living in South Florida.

Methods: This cross‐sectional study included 80 young adults, males (n=42) and females (n=38), ages 18‐36 years recruited from Florida International University campus. Serum 25‐hydroxy vitamin D and TC were measured by ELISA method. Systolic and diastolic BPs were measured by an automated monitor. Weekly total sun exposure and daily vitamin D intake were calculated from validated sun exposure and food frequency questionnaires respectively. A multi‐linear regression model was run, adjusting for age, gender, body mass index, systolic BP, daily vitamin D intake, total weekly sun exposure and tobacco use.

Results: Total cholesterol (P= 0.020) and diastolic blood pressure (P= 0.025) were negatively associated with serum 25‐hydroxy vitamin D. This association remained significant after controlling for covariates (B= ‐0.074, P=.045 for total cholesterol, B= ‐0.421, P= ‐0.018 for diastolic blood pressure). The model explained 18.4% of the variation in serum 25‐hydroxy vitamin D.

Conclusion: The results of this study suggest that high total blood cholesterol and diastolic blood pressure may be associated to low serum 25‐hydroxy vitamin D level. Interventions are needed to confirm these results.

Grant Funding Source: Supported by NIH/NIDDK sponsored grant.
Vitamin D deficiency impairs quadriceps strength without modulating inflammatory cytokines in patients with knee osteoarthritis (1041.2)
Authors: Barker Tyler , Henriksen Vanessa , Rogers Victoria , Aguirre Dale , Trawick Roy , Rasmussen G. , Momberger Nathan
Published: June 2021

The objective of this study was to test the hypothesis that vitamin D deficiency exacerbates quadriceps weakness and increases serum cytokines in patients with knee osteoarthritis (OA). Patients (<60 y) with unilateral knee pain, radiographic evidence of knee OA, and quadriceps weakness participated in this study. Patients were excluded if they presented other co‐morbidities or the use of supplements. Serum 25(OH)D concentrations categorized patients as vitamin D deficient (n=17; 蠄20 ng/mL), insufficient (n=21; 20‐29 ng/mL), or sufficient (n=18; 蠅30 ng/mL) after completing all data collection and measurements. Single‐leg strength (1 set x 6 repetitions of concentric knee extension and flexion contractions at 60°/s on each leg) and blood cytokine, carotene (α and β), lutein, zeaxanthin, ascorbic acid, uric acid, and tocopherol (α and γ) concentrations were measured. Knee extension (quadriceps) torque, power, work, and deceleration were impaired (all p<0.05) with vitamin D deficiency. Deficiency did not increase circulating cytokines but associated with an increase in blood uric acid and γ‐tocopherol concentrations (both p<0.05). In conclusion, vitamin D deficiency is detrimental to quadriceps function in patients with knee OA. Future studies are encouraged to examine other anti‐inflammatory agents, such as γ‐tocopherol, when investigating the role of vitamin D status on inflammatory cytokines.
A randomized controlled trial of vitamin D supplementation in pregnancy: effects on vitamin D status and clinical outcomes (1041.5)
Authors: Zerofsky Melissa , Jacoby Bryon , Stephensen Charles
Published: June 2021

Vitamin D status in pregnancy is associated with health outcomes for mother and infant. The current recommendation for vitamin D intake during pregnancy may be insufficient to raise serum 25‐hydroxyvitamin D (25OHD) to optimal levels. This study evaluated the effect of two treatment regimens on vitamin D status and clinical outcomes. 57 pregnant women at risk for low 25OHD were randomized to treatment (2,000 IU vitamin D3/d) or control (400 IU/d) from <20 wk gestation until delivery. 51 women completed the study. Plasma vitamin D metabolites were measured at <20 wk, 26 wk, and 36 wk gestation. 25OHD was significantly greater in the treatment group at both 26 (104.515.7 vs. 86.320.2 nmol/L) and 36 wk (115.820.1 vs. 85.618.0 nmol/L; p<0.05). The treatment group had a larger change over pregnancy than the control (p<0.0001), and a significantly greater proportion achieved a 25OHD >75 nmol/L by 36 wk (96% vs. 73.1%; p=0.02). Plasma 1,25‐dihydroxyvitamin D increased more in the treatment than the control group between baseline and 26 wk (p<0.01) and 36 wk (p<0.01). Maternal clinical outcomes, including blood pressure and glucose tolerance, did not differ between groups. Infants born to mothers in the treatment group had a marginally (p=0.06) greater birth weight (3,668431 g) than those in the control group (3,379557 g). A daily dose of 2,000 IU is effective at improving vitamin D status in pregnant women.

Grant Funding Source: Supported by UC Davis CTSC Vision Grant, UL1 TR000002 and USDA‐ARS
Carcinoembryonic cell adhesion molecule 6 as a therapeutic target for breast cancer (1048.2)
Authors: Cheng Tsai‐Mu , Liu Yun‐Ru , Chang Chia‐Ching , Chu Hsueh‐Liang , Shieh Dar‐Bin , Su Li‐Jen , Yen Yun
Published: June 2021

Breast cancer is the most frequently diagnosed cancer and the second leading cause of cancer deaths among women. Carcinoembryonic antigen‐related cell adhesion molecule 6 (CEACAM6) is an intercellular adhesion molecule that is overexpressed in several malignancy cancers, including pancreatic, lung, colon, breast, and gastric cancers, is associated with cancer invasion metastasis, tumourigenesis, and drug resistance. We targeted CEACAM6 by an anti‐CEACAM6 single domain antibody (sdAb) or small hairpin RNA (shRNA) to neutralize CEACAM6 functional activity for breast cancer. Inhibition of CEACAM6 functional activity will decrease cultured breast cancer cells proliferation, MMP‐2 and MMP‐9 secretion, migration and invasion in the bio‐coated Matrigel invasion chambers, monocyte infiltration trough 8 μm pores of transwell inserts, and breast cancer cell induced HUVECs angiongenesis in a growth factor reduced membrane matrix assay. According to FITC‐conjugated Annexin V and propidium iodide applied flow cytometric analysis, anti‐CEACAM6 sdAb induced breast cancer cell death through the late stage apoptosis. In addition, our mRNA chip analysis shown the therapeutic effect of sdAb treatment was through regulating 48 apoptosis related gene. According, neutralizing CEACAM6 function activity is a potential therapeutic target for CEACAM6 expressed breast cancer.
Arginase 1 regulates SIRT1 activity and endothelial senescence in diabetes (1051.1)
Authors: Toque Haroldo , Bhatta Anil , Yao Lin , Caldwell Ruth , Caldwell R.
Published: June 2021

Premature senescence in endothelial cells (EC) has emerged as a factor in vascular endothelial dysfunction (VED) during diabetes and other cardiovascular diseases. Diabetes‐induced VED involves elevated arginase (ARG) activity. Reduced levels/activity of sirtuin 1 (SIRT1, a histone deacetylase essential for EC self‐renewal and resistance to stressors) have been linked to premature senescence. We examined the roles of ARG and SIRT1 in diabetes‐induced premature EC senescence by studies using EC specific‐ARG1 knockout (KO) mice and bovine aortic and human coronary EC and WT mouse aorta transduced with ARG1 cDNA (24 hr) or exposed to high glucose (HG, 25 mM, 72 hr). We observed that diabetic or ARG1 transduced aorta exhibited premature EC senescence [measured as senescence‐associated β‐galactosidase (SA‐β‐gal) activity], decreases in SIRT1 activity/expression and VED accompanied by increases in ARG activity and reduced NO production. These effects were not seen in diabetic KO mice with EC specific deletion of ARG1. Furthermore, EC transduced with ARG1 or treated with HG showed marked decreases in SIRT1 activity and NO production which were accompanied by significant increases in activity of ARG and SA‐β‐gal. Hence, reducing ARG1 activity/expression preserves SIRT1 levels, maintains NO production and prevents premature EC senescence and thus has therapeutic potential in attenuating diabetes‐induced VED.

Grant Funding Source: AHA Scientific Development Grant
Expression and localization of alpha‐1 adrenergic receptors in aorta of diabetic Wistar Kyoto and spontaneously hypertensive rats (1051.18)
Authors: Rodriguez Jessica , Reséndiz‐Albor Aldo , Rosales‐Cruz Erika , Hong Enrique , Villafaña Santiago
Published: June 2021

Hypertension and diabetes are multifactorial diseases that frequently coexist and exacerbate each another. During the development of diabetes the impairment of noradrenergic system has been reported especially to the response mediated by alpha‐1 receptors. Although their participation in the development of cardiovascular complications is still controversial some studies have found increased or diminished response to the vasoconstrictive effect of noradrenaline in a time‐dependent manner of diabetes. Thus, the aim of this work was to investigate possible changes in the expression or localization of alpha‐1 adrenergic receptors (alpha‐1A and alpha‐1D) in aorta of rat after 4 weeks of the onset of diabetes. In order to be able to examine the expression of alpha‐1 adrenergic receptors immunofluorescence procedure was performed in túnica intima and media of hystological sections of aorta. Fluorescence was detected by a confocal microscopy. Our results suggest that both receptors showed a higher density in tunica intima and tunica media of SHR aorta compared with WKY rats. Whereas 4 weeks diabetes increases the expression of the receptors dependently of the aortic región and the receptor subtype wherein expression of the alpha‐1A receptor is increased in the intima while the alpha‐1D receptor in tunica media. In conclusion early diabetes modifies the expression of alpha‐1 adrenergic receptors in aorta of both WKY and SHR strains in a different way.

Grant Funding Source: CONACYT CB‐2012‐01‐183660/ COFAA
Positive KCa channel gating modulators with selectivity for KCa3.1 (1057.6)
Authors: Brown Brandon , Coleman Nichole , Oliván‐Viguera Aida , Köhler Ralf , Wulff Heike
Published: June 2021

Intermediate and small‐conductance Ca2+ activated K+ (KCa) channels regulate membrane excitability and Ca2+ signaling. Pharmacological activation of these voltage‐independent channels has therefore been suggested for the treatment of various diseases. While KCa2 activators can potentially reduce neuronal excitability in CNS disorders, selective KCa3.1 activators could be useful as endothelial targeted antihypertensives. KCa channels are gated by Ca2+ binding to a constitutively associated calmodulin at the C‐terminus. Recent work by Zhang et al. (Zheng M. 2012 Nat Commun 3:1021) demonstrated that the benzimidazolone 1‐EBIO binds at the interface of the CAM N‐lobe and the calmodulin binding domain (CAMBD) of KCa2.2. Mutations of A477V/L480M and A477I in KCa2.2 were shown to increase and decrease potency of 1‐EBIO, respectively. Here we report that the same mutations (A625V/L628M and A625I) in KCa2.3 also alter the potency of KCa2/3 activators of the benzothiazole class such as SKA‐31. However, like 1‐EBIO and NS309, SKA‐31 suffers from a lack of selectivity between KCa3.1 and KCa2 channels. Via a structure activity relationship study we have optimized the pharmacophore of SKA‐31 and have now identified several compounds that display 40 to 160‐fold selectivity for KCa3.1 over KCa2.3. SKA‐121, a compound generated through an isosteric replacement approach, lowers blood pressure in mice as determined by telemetry without exerting KCa2 channel mediated effects on heart rate.

Grant Funding Source: Supported by a NIGMS funded T32GM099608
Novel GABAA‐rho1 interactions with acid sensing ion channel ligands (1059.3)
Authors: Snell Heather , Gonzales Eric
Published: June 2021

γ‐ amino butyric acid (GABA) is the major inhibitory neurotransmitter in the vertebrate brain, and targets the ionotropic GABAA receptors. GABAC, or GABAA‐rho, is a subclass of GABAA receptors composed entirely of rho (ρ) subunits and are located on the axonal terminal of retinal bipolar cells, where it not only exhibits a tonic inhibitory current, but also regulates the GABA‐A and other GABAA‐rho synaptic currents (Jones et al 2011). GABAA‐rho exhibits unique properties, such as insensitivity to select antagonists of the heteromeric GABAA receptors (Korpi et al., 2002). A group of ligands, which possess a guanidine group, have been shown to influence GABAA receptors. These compounds, such as (S)‐2‐Guanidinopropionic acid and guanidine acetic acid were competitive antagonists for the GABAA‐rho1 receptor. Other guanidine compounds that are acid sensing ion channel (ASIC) ligands, might also exhibit unique effects on the GABAA‐rho1 receptor. We hypothesize that these ASIC ligands will exhibit unique intrinsic activities on the GABAA‐rho1 receptor, which is different from that of the heteromeric GABAA receptor. The human GABAA‐rho1 receptors were expressed in HEK‐293T cells, and activity was analyzed using whole cell patch‐clamp electrophysiology. When co‐applied with GABA and compared to the GABA concentration profile, one ligand was found to decrease the maximal response, with no change in the GABA EC50. This indicates characteristics of non‐competitive inhibition, while a different ligand shifted the GABA EC50 to lower GABA concentrations, suggesting this guanidine compound is a positive allosteric modulator. When applied alone, it failed to directly activate GABA rho1 receptors. These contrasting effects suggest that these ligands act at two binding sites within the GABAA‐rho architecture. Future experiments will focus on additional characterization of these novel effects on GABA rho receptors and offer a novel chemical structure to design novel GABAA‐rho therapeutics.

Grant Funding Source: Supported by: Welch Foundation Grant No: BK‐1736, NIH training grant: NBA T32 AG020494
BPC 157 antagonized the general anesthetic potency of thiopental and reduced prolongation of anesthesia time induced by L‐NAME/thiopental combination (1061.3)
Authors: Zemba Mladen , Zemba Cilic Andrea , Balenovic Igor , Cilic Matija , Radic Bozo , Suran Jelena , Drmic Ivan , Stambolija Vasilije , Murselovic Tamara , Katancic Holjevac Jadranka , Uzun Sandra , Djuzel Viktor , Seiwerth Sven , Sikiric Predrag
Published: June 2021

We hypothesized the general anesthetic thiopental effect depending on NO‐related mechanisms, consequently counteracted by stable gastric pentadecapeptide BPC 157.

(i) To determine the general anesthetic effect of thiopental and possible counteraction depending on BPC 157 administration, all rats received intraperitoneally thiopental (20, 30, 40 mg/kg) while medication (BPC 157 (10 μg/kg, 10 ng/kg, 10 pg/kg) was given intraperitoneally at 5 min before thiopental. (ii) To determine NO‐related mechanisms, all rats received intraperitoneally thiopental 40 mg/kg while BPC 157 (10 μg/kg), L‐NAME (10 mg/kg), L‐arginine (30 mg/kg) were applied alone and/or combined. BPC 157 was given at 25 min before thiopental while L‐NAME, L‐arginine, alone and/or combined, were applied at 20 min before thiopental.

(i) BPC 157 (10 ng/kg and 10 μg/kg), caused significant antagonism of general anesthesia produced by thiopental with a parallel shift of the dose‐response curve to the right. (ii) L‐NAME. Thiopental‐induced anesthesia duration was tripled. L‐arginine. Active only given with L‐NAME or BPC 157: habitual thiopental anesthesia time not influenced; potentiating effects of L‐NAME lessened, not abolished; shortening effect of BPC 157: abolished. BPC 157 and L‐NAME. Potentiating effects of L‐NAME was abolished. L‐NAME and L‐arginine and BPC 157. L‐NAME+L‐arginine+BPC 157 rats exhibited values close to those in BPC 157 rats.

Thiopental general anaesthesia is simultaneously manipulated in both ways with NO‐system activity modulation, L‐NAME (prolongation) and BPC 157 (shortening/counteraction). BPC 157 and L‐arginine might serve two NO‐system pathways, more or less active, alternatively activated.

Grant Funding Source: Supported by Grant 108‐ 1083570‐3635, Croatia
Tumor‐targeting and anti‐tumor activity of anti‐claudin‐3/‐4 antibody (1062.7)
Authors: Kimura Yuka , Li Xiangru , Iida Manami , Tada Minoru , Ishii Akiko , Watari Akihiro , Fukasawa Masayoshi , Yagi Kiyohito , Kondoh Masuo
Published: June 2021

Claudins (CLDNs) comprise a tetra‐transmembrane protein family with ~20 members, which are located in the lateral membrane between adjacent epithelial cells. Because of loss of cellular polarity during malignant transformation, CLDN often migrates from the lateral membrane and becomes exposed on epithelial cell surfaces. CLDN3 and CLDN4 are targets for tumor therapy because they are overexpressed in various malignant tumors. We previously optimized a protocol for immunization with CLDN and developed 8 rat anti‐CLDN4 monoclonal antibodies, one of which recognized both CLDN3 and CLDN4. In this study, we focused on the CLDN3/4 antibody. The CLDN3/4 antibody showed high affinity for CLDN4 (KD value: 4.3 nM). We fluorescently labeled the CLDN3/4 antibody and intravenously injected it into mice bearing tumors (human colon cancer Lovo cells, or human gastric cancer MKN74 cells). The CLDN3/4 antibody accumulated in tumor tissues and attenuated tumor growth without apparent adverse effects in the Lovo‐ and MKN74‐bearing mice. Furthermore, CLDN3/4 antibody activated the Fcγ receptor in vitro. Therefore, the CLDN3/4 antibody may exert its anti‐tumor activity through antibody‐dependent cellular toxicity. These findings suggest that the CLDN3/4 antibody is a candidate anti‐tumor agent.

Grant Funding Source: supported by the Ministry of Health, Labor, and Welfare of Japan
Cerebral blood flow regulation during blood loss compared to lower body negative pressure in humans (1068.9)
Authors: Barnes Jill , Johnson Blair , Convertino Victor , Joyner Michael , Rickards Caroline
Published: June 2021

Lower body negative pressure (LBNP) is often used to simulate blood loss in humans. However, it is unknown if the effects of actual blood loss (BL) on cerebral blood flow regulation is analogous to simulated blood loss during LBNP. Nine healthy men (32±6 y) were studied at baseline, during 3 levels of LBNP (5 min each at ‐15, ‐30, ‐45 mmHg), and during 3 levels of blood loss (5 min each at 333, 667, 1000 mL). LBNP and BL conditions were randomized. Intra‐arterial mean arterial pressure (MAP) was similar during LBNP compared with BL (p>0.20). Central venous pressure (CVP; 2.8±0.7 vs. 4.0±0.8, 1.2±0.6 vs. 3.5±0.8, 0.2±0.9 vs. 2.1±0.9 mmHg for level 1, 2, and 3; p<0.01) and stroke volume (71±4 vs. 80±3, 60±3 vs. 74±3, 51±2 vs. 68±4 mL for level 1, 2, and 3; p<0.01) were lower during LBNP compared with BL. Despite differences in CVP, middle cerebral artery velocity (MCAv) and cerebrovascular conductance index (CVCi) were similar between LBNP and BL at each level (MCAv at level 3: 62±6 vs. 66±5 cm/s; p=0.37; CVCi at level 3: 0.72±0.05 vs. 0.73±0.05 cm/s/mmHg; p=0.29). Furthermore, the slope of the relationship between MAP and MCAv was not different between LBNP and BL (1.06 vs. 1.07 cm/s/mmHg). Our results suggest that the cerebral hemodynamic responses to LBNP are similar to actual blood loss.

Grant Funding Source: Supported by U.S. Army MRMC Combat Casualty Care Research Program Grant # W81XWH‐11‐1‐0823
Dynamic and static factors predicting vasomotor reactivity in forced hypocapnia (1070.8)
Authors: Müller Martin , Österreich Mareike
Published: June 2021

Objectives: In mechanically ventilated patients intensive hyperventilation can be detrimental. Using transfer function (TF) estimates we were interested in which Parameter could be used for monitoring autoregulation in this condition.

Methods: 18 healthy persons (9 women; mean age 32 +‐9 years) underwent simultaneously recordings of blood flow velocity (BFV) by means of transcranial Doppler sonography, blood pressure and endtidal pCO2 (ETpCO2) during 4‐5 minute periods of normoventilation and voluntary hyperventilation. Breath holding (BH) as long as possible started immediately at the end of hyperventilation. TF and Coherence were calculated using Welsh's periodogram method. Mean values were calculated for phase and gain in the very low frequency (vlf) range (0.02‐0.07 Hz), low frequency (lf) range (0.07‐0.15 Hz) and high frequency (hf) range (0.15‐0.40 Hz). VMR was indicated by: 1. VMR% = [BFVnormov‐(BFVbh‐BFVhyperv)/BFVnormov]* 100; 2. BHI= VMR% divided by time of BH.

Results: Mean BP was 83±10 mm HG in normoventilation (ETpCO2 0.53 ±0.05 Vol%), 84±11 in hyperventilation (ETpCO2 0.36±0.05) and 82±14 at BH release (ETpCO 0.56 ±0.06); BFV was 59±19 cm/sec, 47±17 and 78±28 cm/sec respectively. Stepwise regression analysis indicated: 1. VMR% was significantly related to ETpCO2 (r=0.50, p<0.01), lf gain (r=0.40, p<0.01) and hf gain (r=0.63, p<0.001) each under hypocapnia; 2. BHI was significantly related to lf phase (r=‐0.58; p=0.01) and lf gain (r=0.63; p<0.001) under hypocapnia and to hf gain (r=0.49; p<0.01) under normocapnia.

Conclusion: VMR% and BHI remained related to blood pressure derived gain function. The negative correlation between phase and BHI may give a first hint why extreme hypocapnia can be detrimental to patients under mechanical ventilation
Inhibition of high salt‐induced activation of NLRP3 inflammasome by mesenchymal stem cell transplantation in the renal medulla in Dahl S rats (1074.7)
Authors: Li Xiaoxue , Zhu Qing , Conley Sabena , Hu Junping , Li Pin‐Lan , Li Ningjun
Published: June 2021

Activation of NALP3 inflammasome has been shown to be involved in various renal pathological conditions. It remains unknown whether renal NALP3 inflammasome activation participates in the abnormal renal response to high salt diet in Dahl salt sensitive rats. In addition, whether inhibition of NALP3 inflammasome activation contributes to the renal protective actions of stem cells is not clear. The present study detected the response of NALP3 inflammasome to high salt intake and determined the effect of mesenchymal stem cell (MSC) transplantation on inflammasome activation. Immunostaining showed that 3 essential components of the NALP3 inflammasome, NALP3, ASC and caspase‐1, were all mainly present in distal tubules and collecting ducts. Interestingly, the renal medullary mRNA and protein levels of these inflammasome components were remarkably increased by 2‐4 fold in response to high salt intake in Dahl S rats, while remained no change in normal rats. High salt‐induced increases of renal medullar inflammasome were significantly blocked by MSC transplantation into the renal medulla compared with control cell in Dahl S rats. These data suggested that high salt‐induced activation of renal medullary inflammasome may contribute to the renal medullary dysfunction in Dahl S rats and that inhibition of inflammasome activation may be one of the mechanisms for the protective effects of stem cells in the renal medulla.

Grant Funding Source: HL089563 and HL106042
Mechanism of inhibition of delayed rectifier K+ current by diphenyl phosphine oxide‐1 in porcine coronary smooth muscle (1079.10)
Authors: Fancher Ibra , Noblet Jillian , Goodwill Adam , Tune Johnathan , Dick Gregory
Published: June 2021

Mechanisms involved in DPO‐1‐induced block of KDR were studied in myocytes isolated from the porcine coronary artery. DPO‐1 inhibited the slowly inactivating KDR in a dose‐dependent manner (IC50 = 212 ± 28 nM). Neither the time constant of activation (+20 mV) nor the time constant of deactivation (‐40 mV) was affected by DPO‐1 (1 μM). However, the fast and slow time constants of inactivation at +20 mV were notably shortened by DPO‐1 (fast = 0.52 ± 0.07 vs. 0.14 ± 0.01 s; slow = 5.8 ± 1.1 vs. 0.6 ± 0.2 s). These results suggest that DPO‐1 preferentially binds to the channel in the open state, producing an open channel block of KDR. As such, if KDR channels were open under resting conditions in vascular smooth muscle, then DPO‐1 should depolarize the membrane potential and cause vasoconstriction. To test this hypothesis and show functional relevance, we determined the effect of DPO‐1 on the membrane potential and coronary blood flow. DPO‐1 depolarized membrane potential from –29 ± 1 to ‐14 ± 3 mV. Further, DPO‐1 reduced coronary artery blood flow at 100 mmHg perfusion pressure from 21 to 14 ml/min. Together, these data indicate that DPO‐1 blocks KDR channels in the open state and that KDR channels in vascular smooth muscle are open under resting conditions. Thus, therapeutic tools that target KDR activity could be used to regulate vascular tone and mechanisms controlling KDR merit further study.

Grant Funding Source: Supported by T32HL090610, T32HL079995, AHA 13POST1681001813, HL092245
Thymoquinone protects against myocardial ischemia‐reperfusion injury via modulation of oxidant generation and nuclear factor‐kappaB‐mediated responses (1080.1)
Authors: El‐Mahdy Mohamed , Ismail Raed , Hemann Craig , El‐Sherbiny Gamal , Abdelghany Tamer , Helal Gouda , Zweier Jay
Published: June 2021

Understanding the mechanisms of natural compounds in modulating disease has moved to the forefront of cardiovascular research. Thymoquinone (TQ), a constituent of the volatile oil derived from Nigella sativa seeds, has demonstrated promising effects against ischemia‐reperfusion (IR) injury of different organs; however, the mechanism of TQ action on myocardial IR injury (m‐IRI) warrants more investigation. Rat hearts were either subjected to 30 min of global ischemia followed by 120 min of reperfusion using a Langendorff system, or to 30 min of left anterior descending coronary artery (LAD) ligation followed by 24h of reperfusion. TQ was infused ex‐vivo for 10 min either pre‐or post‐ischemia or IP‐injected for seven days prior to the surgery. TQ significantly reduced reactive oxygen species (ROS) generation, infarct size and apoptosis, and markedly enhanced coronary flow and ventricular function of the ischemic hearts. TQ attenuated the IR‐induced up‐regulation of SAPK/JNK, myocardial nuclear factor (NF)‐kappaB, TNF‐α, and P38‐MAPK expression and increased the Bcl‐2/Bax ratio. The data indicate that TQ inhibited ROS generation‐induced NF‐kappaB which led to inhibition of pro‐inflammatory cytokines and apoptosis, protecting the heart against IR injury. This study presents a new molecular mechanism of action and efficacy of TQ with promise as a natural compound in the management of m‐IRI.

Grant Funding Source: NIH# 00000784 & Egyptian Government