bluuebunny/crossref_metadata_2025 huggingface dataset:
Creation of a mobile‐phone application for assessing supermarket offerings for a Guatemalan‐based exploration of the Nutrient Environment Measurement Survey‐Stores (255.1)
Authors: Kanter Rebecca , Alvey Jeniece , Fuentes Déborah
Published: June 2021

Background: Consumer nutrition environment measures are important to understanding the food environment, which affects individual diet. A paper version of the nutrition environment measures survey for supermarkets (NEMS‐S) was modified for use in Guatemala. However, a paper survey is not an inconspicuous data collection method and is not well equipped for multiple languages in the same version.

Objective: To design, pilot test, and validate the paper NEMS‐S for Guatemala in the form of a mobile phone application (mobile app).

Methods: CommCare, a free and open‐source software application, was used to design the NEMS‐S for Guatemala in the form of a mobile app. Two raters tested the mobile app in one Guatemalan supermarket. Both the inter‐rater and test‐retest reliability of the mobile app was determined using percent agreement and Cohen’s Kappa score; and compared to the paper version.

Results: The inter‐rater reliability was very high between the paper survey and the mobile app (Cohen’s kappa > 0.90). Test‐retest reliability ranged from Kappa 0.78‐0.91. With experience, survey completion‐time using the mobile app was 5 min less than the paper form (35 vs 40 min).

Conclusions: The NEMS‐S mobile app provides for more rapid data collection, with equivalent reliability and validity of the NEMS‐S paper version. And has advantages over a paper‐based survey of multiple language capability and concomitant data entry.
Residential property values: a novel GIS‐based metric to characterize obesogenic environments (255.8)
Authors: Aggarwal Anju , Rehm Colin , Cohen‐Cline Hannah , Hurvitz Phil , Moudon Anne , Drewnowski Adam
Published: June 2021

Objective: To determine whether multiple attributes of obesogenic environments can be captured by the single objective measure of residential property values. Methods: The Seattle Obesity Study (SOS) was a cross‐sectional study on socioeconomic disparities in diets and health. A telephone survey of a geographically distributed sample of 2,001 King County adults was conducted in 2008‐09. Home addresses were geocoded. Residential property values at tax parcel level were obtained from King County tax assessor. Mean residential property values within a 10‐minute walk (833m buffer) were calculated. Perceptions toward neighborhood social and physical environment were collected by self‐report. Multivariable linear regression analyses examined the associations among multiple attributes of the neighborhood environment, property values and body mass index (BMI). Results: Perceived neighborhood attributes such as crime, heavy traffic and proximity to bars, liquor stores and fast foods were all associated with lower property values. By contrast, living in neighborhoods that were perceived as safe, quiet, clean and attractive was associated with higher property values. Higher property values were in turn associated with lower BMIs. The observed associations between perceived neighborhood characteristics and BMI were largely attenuated after accounting for property values. Conclusion: Environments, described as obesogenic in past studies, were consistently associated with lower property values. Further research, in additional locations, needs to explore whether residential property values act as a proxy for other neighborhood characteristics and socioeconomic status.

Grant Funding Source: NIH grants P20 RR020774‐03, R01 DK076608‐04 and R21 DK020774
Differences among pulse grains and cereal grains in dietary fiber (259.5)
Authors: Thompson Henry , Echeverria Dimas , Brick Mark
Published: June 2021

Pulses, i.e., dry beans, dry peas, chickpeas, and lentils, are grain legumes that serve as staple food crops for populations around the world. Since ancient times, pulses have been combined with cereal grains, primarily wheat, rice, or corn, or tubers such as potato or cassava to provide a balanced source of protein and carbohydrate and a nutritionally adequate diet when combined with seed oils, fruit and vegetables. In this setting, the established ratio of cereal grains to pulses to meet nutritional requirements is 2:1; however, over the last 60 years both production and consumption patterns, globally, have changed to 8:1. One consequence of these changes has been on a marked but largely unappreciated shift in dietary carbohydrate quality. This presentation focuses on differences among pulse crops in their amount and type of dietary fiber and how this relates to the amounts and types of dietary fiber provided by various cereal grains. Data will be reported using the AOAC 2011.25 Integrated Total Dietary Fiber Assay to quantify dietary fiber and oligosaccharide content. These data will be considered in the context of the pandemic of overweight and obesity that has swept the globe over the last 60 years since there has been virtually no scrutiny of the potential association between the decline in pulse consumption and the global rise in obesity and its associated metabolic disorders.

Grant Funding Source: PHS R01‐ CA172375 and American Pulse Association
Maternal food insecurity, anemia, and social support are independently associated with prenatal depression among HIV‐positive and ‐negative women attending antenatal services in northern Uganda (268.2)
Authors: Natamba Barnabas , Achan Jane , Oyok Thomas , Mou Sophie , Gosh Shibani , Stoltzfus Rebecca , Griffiths Jeffrey , Young Sera
Published: June 2021

Introduction: Limited data exist on prenatal depression in in high HIV burden resource‐poor settings.

Population: 403 pregnant clinic attendees at Gulu Hospital, Uganda. HIV (+) women were oversampled in a ratio of 1 HIV (+): 2 HIV (‐).

Methods. Depression was assessed using the Center for Epidemiological Studies‐Depression (CESD) scale. Maternal food insecurity (FI) was evaluated using an individually focused Food Insecurity Access Scale, anemia using a Hemocue® hemoglobinometer, and social support using the 10 item version of the Duke‐UNC functional social support instrument. We performed bivariate and multivariate analyses to identify variables associated with CESD scores keeping those with p <0.05 as well as theoretically or contextually important variables in the final model.

Results: CESD scores ranged from 0‐57 (mean 20 + 13.2SD). 62.3% of study women had a major depressive episode (CESD score 蠅15). While being HIV (+) was linked to higher CESD scores in bivariate analyses, in multivariate linear regression maternal food insecurity (β=+0.663) and age (β=+0.227) were the only factors associated with higher CESD scores. Maternal social support (β=‐1.209) and higher hemoglobin (β=‐0.927) were linked to lower CESD scores.

Conclusion: Interventions targeting depressed pregnant Ugandan women should better understand how age, food insecurity, anemia, and social support influence prenatal depression.

Grant Funding Source: Supported by USAID Feed the Future grant to Tufts University
The quality of diets reported by Expanded Food and Nutrition Education Program participants in the Mountain Region in 2011 when exiting the program was higher than when entering (273.8)
Authors: Luick Bret , Guenther Patricia
Published: June 2021

EFNEP is USDA’s largest nutrition education program for low income people and receives about $68,000,000 in funding annually. Evaluations have consistently shown positive results, but the 24‐hr recalls of dietary intake have not been widely used. Our objective was to compare the quality of diets reported by participants when entering the program with those reported when exiting. Because the 2005 Dietary Guidelines were in effect when the curriculum was developed, the Healthy Eating Index (HEI)‐2005 was selected as the quality indicator. Recalls were collected by trained paraprofessionals using a standard format and were coded using the USDA food coding system. This study was a secondary analysis of data collected from 3,367 female participants in the eight states of the Mountain region from October 2010 through September 2011. We calculated mean HEI‐2005 scores for entry and exit recalls. Significant differences were determined using the Wilcoxon signed‐rank test. The majority of participants were Hispanic. At entry the mean total HEI‐2005 score was 49.1 (out of a possible 100); and at exit, 55.2. Nine of the 12 component scores also improved significantly while Sodium worsened. Because of the self‐reported data, lack of biomarkers, and no control group, it is difficult to conclude that diet quality improved; however, it is likely that nutrition knowledge improved and led to short‐term improvement in diet quality.
Tracking chromatin signatures in individual cells to trace vascular smooth muscle cell lineage in human atherosclerotic lesion (278.4)
Authors: Gomez Delphine , Shankman Laura , Owens Gary
Published: June 2021

Atherosclerosis is a chronic disease of the arterial wall responsible for nearly 50% of deaths in developed countries. There is a strong correlation between ratio of smooth muscle cells (SMC) to macrophages and stability of atherosclerotic lesions; a high macrophage to SMC content associated with high risk of plaque rupture. Herein, we reevaluated the contribution of SMC within lesions that was underestimated due to lack of identification of SMC undergoing phenotypic switching (i.e loss of SMC marker gene expression). Using a SMC‐lineage tracing mouse, we show that traditional staining for SMC is unable to detect 86% of the SMC population within lesions of Apoe‐/‐ mice and that 25% of macrophages identified by traditional macrophage markers are of SMC origin. To validate these data in human, we developed a novel method of detection of histone modifications on gene loci in single cells using tissue sections. Our assay was based on studies showing that the histone modification H3K4dime on the SMC marker genes is a specific and stable epigenetic marker of the SMC lineage in vitro. First, H3K4dime on the SMC marker genes (PLA+ cells) was strictly restricted to SMC lineage in vivo. Then, we identified PLA+/macrophage markers+ cells in human lesions. In conclusion, we rigorously identified lesion cells of SMC origin and we provide exciting evidence that a fraction of macrophage‐like cells in human lesions are derived from SMC.

Grant Funding Source: American Heart Association
The fossil evidence for the evolution of human infant helplessness, complicated labor and big babies (335.3)
Authors: Rosenberg Karen
Published: June 2021

The human fossil record is much richer today than it was when the obstetrical dilemma was first proposed to explain the conflicting selective constraints that result when bipeds give birth to large‐brained babies. An analysis of the morphology of several important discoveries of fossilized pelvic material from Gona in Ethiopia and Malapa in South Africa, demonstrate that a broad pelvis such as characterizes humans today did not result from expansion of the birth canal that coincided with encephalization during the last two million years, but was the primitive condition for hominins, existing in australopithecines, Homo erectus and all later humans. However, encephalization did select for further changes in the pelvis as well as changes in infant development and parental behavior. The complicated way that humans give birth and the maturational status of neonates are a result of that evolutionary history and the resulting constraints that include: bipedalism, our large brained newborns, the large size of those newborns relative to the size of the mothers who give birth to them, the helpless state in which these large babies arrive and the prolonged period of infant dependency. This expanded fossil record has implications for an understanding of the way we give birth and care for our newborn infants.
Changes in tumor microenvironments in high‐fat diet‐fed mice: role of macrophages and adipocytes (37.5)
Authors: Park Jung , Cho Han Jin , Jung Jae In , Kwon Gyoo Taik , Lee Ki Won
Published: June 2021

Using several mouse tumor models, we have shown that the chronic consumption of a high‐fat diet (HFD) stimulates solid tumor growth and metastasis and thereby reduces the survival rate. The present study examined the mechanisms by which HFD stimulates tumor promotion. Angiogenesis and lymphangiogenesis in tumor tissues and lymph node (LN)s are markedly increased in HFD‐fed mice. HFD feeding increases adipocytes and M2‐macrophage (M2‐MΦ)s in tumor tissues and M2‐MΦs in adipose tissues of tumor‐bearing mice. Results from in vivo and in vitro co‐culture studies revealed that the crosstalk between adipocytes, tumor cells, M2‐MΦs, and/or lymphatic endothelial cell (LEC)s stimulates the production of several factors which stimulate tumor cell/monocyte migration, angiogenesis, and lymphangiogenesis via endocrine and paracrine mechanisms. In HFD‐fed mice, increased chemokine ligands in LECs recruit tumor cells expressing high levels of their receptors, thereby increasing LN metastasis. Our results indicate that increased adipocytes in HFD‐fed mice play important roles in the recruitment of M2‐MΦs in tumor tissues and lymphangiogenesis as well as tumor cell migration, thereby stimulating tumor metastasis.
An acute non‐lethal method to test leptin responsiveness in rats (371.3)
Authors: Desai Bhavna , Harris Ruth
Published: June 2021

Continuous subcutaneous administration of leptin normalizes blood glucose levels in rodent models of type I and type II diabetes independent of changes in food intake, body weight and plasma insulin. We tested whether an acute intravenous leptin infusion changed blood glucose in normal and diet‐induced leptin resistant rats and determined if this could be used as a marker of leptin sensitivity. Leptin responsive, chow fed male Sprague Dawley rats and leptin resistant rats fed chow, lard and 30% sucrose or chow and 30% sucrose were fitted with jugular vein catheters. They were infused with either saline for 32 min, low dose (LD) leptin (1.9µg/kg/min) followed by high dose (HD) leptin (3.75µg/kg/min) for 16 min each, or only HD leptin for 16 min. Infusion of either LD leptin or HD leptin alone did not change blood glucose. Infusion of LD leptin followed by HD leptin decreased blood glucose in leptin responsive, but not leptin resistant rats. Leptin infusion increased serum leptin in all groups, but had no effect on serum insulin, glucagon, 18 hour weight gain or 18 hour energy intake in any rat. The leptin‐induced decrease in blood glucose correlated with leptin responsiveness measured as a leptin‐induced inhibition of food intake. This effect is consistent in control rats and absent in two rat models of leptin resistance. Thus, this acute non‐lethal test is a novel method of testing leptin responsiveness in rats.

Grant Funding Source: Supported by NIH DK 53903
Exploring the use of text messages for increasing nutrition knowledge and improving dietary practices in teen athletes (379.6)
Authors: Bilgiç Pelin , Tamer Funda , Yabanci Nurcan , Hamamcilar Osman , Bilgiç S. Cem , Hongu Nobuko
Published: June 2021

The nutrition knowledge and dietary practices of teen athletes is important for both their sports performance and overall health. Our approach to improving health communication with teen athletes takes advantage of the increasing prevalence of mobile phone and SMS messaging. This study consists of two parts: 1) conducting knowledge questions regarding nutrition and 2) developing text messages to support teen athletes. For the first part, 129 male high‐school elite wrestlers with a mean age of 16 (15‐17) years old and mean body weight of 66.6kg±15.9 (SD) were asked about their dietary practices along with general nutrition and sports nutrition knowledge. Athletes answered 59.7% of the general nutrition and 63.9% of the sports nutrition questions correctly. 58 % of the athletes believed that vitamins and minerals maintain their energy. 33.4% of the athletes answered incorrectly to the question asking which mineral promotes healthy bones and teeth. 31.8% of the athletes were skipping meals (36.6%, skipping breakfast; 46.3%, skipping lunch). 24.8% of the athletes were using ergogenic aids, and 35.0% and 15.0% were using vitamin mineral mixtures and protein powders respectively. For the second part of the study, over 100 messages were created by college students by obtaining information from popular consumer magazines or websites, health organization websites, and literature review. These messages were refined by researchers for delivery through SMS. Through a participatory process, educational text messages will be tested based on how teen athletes respond to these text messages, and their suggestions for developing additional topics of interest will be considered.
Adaptation and evaluation of the WillTry tool to assess willingness to try fruits and vegetables among children 3‐11y in Guam (380.6)
Authors: Aflague Tanisha , Leon Guerrero Rachael , Boushey Carol
Published: June 2021

Children’s fruit & vegetable (FV) intake is mediated by a preference for FV or a willingness to try FV. Our aim was to adapt the validated WillTry (Thomson et al Appetite 2010) for interventions in Guam among children, 3‐11 y. Adaptations included novel and common FV unique to Guam. Children were shown images matching the 14 food questions during a one‐on‐one interview and a 2nd interview 3‐72 hrs later for reliability. Factor analyses determined components of willingness to try FV. We analyzed psychometric properties and reliability. Sixty‐five children from 2 camps completed the first interview and 64 did the second. Factor analyses revealed 3 dimensions, i.e., local novel (guava, breadfruit, eggplant, sweet sop, star apple, taro leaves), local common (carrot, papaya, long beans, salad greens), and imported (apple, canned peaches, canned corn). All but the imported scale had sufficient internal consistency (Cronbach’s α>0.69). Each food scale had substantial reliability (ICC>0.76). There were no significant differences by age, sex, or camp for any scale. Mean scores were 2.1 (local novel), 2.4 (local common), and 2.7 (imported) and all were significantly different. The adapted WillTry was culturally relevant and possessed similar psychometric properties as the original. An unexpected finding was the tool’s potential for documenting shifts from local to imported FV, a concern of the nutrition transition.

Grant Funding Source: Supported by NCI 1U01CA130784‐01; USDA NIFA no 2011‐68001‐30335
The SORT1 risk allele is associated with exaggerated postprandial lipaemia in young adults (383.5)
Authors: Connors Kimberly , Gnatiuk Elizabeth , Shearer Jane , Hoffman Eric , Barfield Whitney , Hittel Dustin
Published: June 2021

Elevations in low‐density lipoproteins (LDL), very‐low density lipoproteins (VLDL) and triglycerides (TG) in the postprandial state have a strong genetic component and recognized atherogenic potential. We sought to determine if there is a difference in postprandial lipaemia following an oral fat tolerance test when stratified by genotype at the SORT1 1p13 locus, the strongest LDL‐cholesterol associated locus identified to date. Thirty, healthy young adults (age=23.57) received a high fat, mixed meal (91 g, 55% kcal from fat). Blood samples drawn at fasting, 2, 4, 8 and 24 hrs postprandial, underwent comprehensive lipoprotein analyses via nuclear magnetic resonance profiling. At fasting, VLDL particles differed based upon SORT1 genotype (TT: 48.81±5.07 nmol/L versus TC/CC: 30.05±2.99 nmol/L, p=0.004). Major, risk allele homozygotes (TT) had significantly elevated area‐under‐the‐curve and mean concentration for VLDL particles, TG and VLDL‐TG (p<0.05). These results indicate increased fasting and postprandial exposure to atherogenic particles in TT homozygotes. This highlights SORT1 as a valuable target for identifying individuals for early, personalized interventions to prevent atherosclerosis.
Gender differences in impact of food stores in neighborhoods on children’s obesity status change: U.S. early childhood longitudinal study (390.4)
Authors: Chen Hsin‐Jen , Wang Youfa
Published: June 2021

Objective: Examine influence of food stores in neighborhoods on children’s weight change by gender. Methods: Height and weight of a national representative sample of US schoolchildren were directly measured at 5th and 8th grades (n=7,090). Information of food store quantities in neighborhoods (supermarkets, convenience stores, takeaway restaurants) in the years of follow‐ups (2004 and 2007) was obtained from the US Zip‐code Business Pattern Data. Children’s weight status and neighborhood environment characteristics were linked via children’s residence zip code. Results: Mixed‐effect models showed that girls living in neighborhoods with 2 supermarkets had lower odds of becoming obese 3 years later (OR=0.44 [95% CI: 0.23‐0.83]) than did those living in neighborhoods without supermarket. Girls living in neighborhoods with more takeaway restaurants had greater odds of becoming obese 3 years later than those living in neighborhoods with 蠄1 takeaway restaurant (OR=4.45 [1.54, 12.83]). Decreased number of supermarkets in neighborhoods was associated with increased odds of obesity for girls (OR=1.68 [1.07, 2.64]). However, there was no significant finding for boys. Conclusions: Girls’ weight status change at the pubertal age, but not boys’, was associated with local food store environments.

Grant Funding Source: NIH (R01DK081335‐02, R01HD064685‐02)
Inhibition of tumor growth by ERK1/2 inhibitors is associated with the induction of interferon‐γ production (397.3)
Authors: Ma Xingzhe , Wang Qixue , Duan Yajun , Han Jihong
Published: June 2021

Inhibition of tumor growth by ERK1/2 inhibitors is associated with the induction of interferon‐γ production

Xingzhe Ma, Qixue Wang, Yajun Duan, Jihong Han

State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin 300071, China

ERK1/2 plays an important role in cell survival, proliferation and differentiation. However, the high ERK1/2 activity has been linked to tumorigenesis. Several ERK1/2 inhibitors are in clinical trials to evaluate their effects on cancers. IFN‐γ is an important cytokine with anti‐tumorigenic function. So far, it has been remained unknown if the anti‐cancer action of ERK1/2 inhibitors is related to their effect on IFN‐γ expression. We previously reported that activation of LXR induces IFN‐γ expression and inhibits tumor growth, and ERK1/2 inhibitors can activate LXR‐targeted gene expression. Herein, we determined if ERK1/2 inhibitors can influence IFN‐γ expression, and the anti‐tumorigenic action of ERK1/2 inhibitors is completed, in part, by their effect on IFN‐γ production. We observed that ERK1/2 inhibitors increased IFN‐γ protein expression which was associated with increased IFN‐γ mRNA expression, promoter activity and formation of LXRE‐nuclear protein complexes. In vivo, U0126 increased mouse serum IFN‐γ levels and IFN‐γ expression in tissues. Animal study also demonstrated that U0126 inhibited inoculated LLC1 tumor growth in wild type but not in IFN‐γ‐/‐ mice indicating the inhibition is associated with IFN‐γ production. Our study suggests that ERK1/2 inhibitor induces IFN‐γ production and the induction can be partially attributed to their anti‐tumorigenic properties.
Human beta‐defensins‐1, ‐2, and ‐3 production by human amniotic epithelial cells in relation to human papillomavirus genotype (397.4)
Authors: Szukiewicz Dariusz , Alkhalayla Habib , Stangret Aleksandra , Szewczyk Grzegorz
Published: June 2021

HPV is common sexually transmitted infection. Based on the oncogenic potential, low‐risk and high‐risk HPV types were distinguished among approximately 40 HPV genotypes identified within the mucosa of the anogenital tract. Epithelial HBDs act as a potent protein adjuvants promoting cellular and humoral immune responses in viral infections. Unlike alpha‐defensins, HBDs are upregulated in HPV infections. We compared strenght of this response in respect to oncogenic (types 16 and 18) and non‐oncogenic (6 and 11) HPVs (groups I and II, respectively). After normal pregnancies (N = 96) HAEC were isolated from the amnions using trypsinization and divided into three groups (including negative controls; group III), according to the results of the vaginal smear real‐time PCR test for detection of HPV DNA. HAEC stimulated with lipopolysaccharide (1μg/ml) were cultured in vitro for 5 days. HBD‐1, HBD‐2, and HBD‐3 were quantified by ELISA in the culture media samples. Observed mean increases for HBDs were 5.7‐, 3.4‐, and 4.6‐fold, compared to the controls. The mean increases in HBDs concentrations were weaker in the oncogenic HPV infection, compared to the non‐oncogenic HPVs, and the differences between group I and II reached statistical significance (p < 0.05) for HBD‐1 and HBD‐3. Thus, HPV‐related HBD overproduction depends on the virus genotype. Subdued response to oncogenic HPVs may be included in their pathomechanism.

Grant Funding Source: Supported by WUM grant: 2M2‐W2‐13.
β‐Catenin and GSK‐3β are targets for regenerative therapies for acetaminophen‐induced acute liver failure (398.4)
Authors: Bhushan Bharat , Walesky Chad , Borude Prachi , Manley Michael , Apte Udayan
Published: June 2021

Acetaminophen (APAP) overdose is the major cause of acute liver failure (ALF) with limited treatment options. Liver regeneration is a critical determinant of survival after APAP overdose and regenerative therapies hold a great potential to treat APAP‐induced ALF. However, mechanisms of regeneration following APAP overdose are not known. We did a comprehensive analysis of pathways involved in liver regeneration using a novel incremental dose model of a “regenerative” dose (300 mg/kg, APAP300) and a “non‐regenerative” dose (600 mg/kg, APAP600) of APAP in mice. Stimulation of regeneration at APAP300 resulted in complete recovery, while inhibited regeneration at APAP600 resulted in delayed recovery and decreased survival. The inhibition of regeneration in APAP600 was due to inhibited cyclinD1 expression leading to delayed cell cycle. Further biochemical and genomic analysis revealed that many known regenerative pathways (e.g. EGFR/c‐Met, STAT3, MAPKs) remain activated even at non‐regenerative dose. However, Wnt/β‐catenin was specifically activated only at the regenerative dose. ChIP analysis revealed increased binding of β‐catenin to cyclinD1 promoter only at APAP300 correlating with higher cyclinD1 induction and liver regeneration. Further, mice overexpressing stable form of β‐catenin exhibited significantly higher regeneration and faster recovery after APAP overdose. Finally, pharmacological inhibition of GSK3β, the upstream regulator of β‐catenin, as late as 4 hr after APAP600 dose resulted in cyclinD1 induction and improved regeneration. These data demonstrate that canonical Wnt signaling pathway is a target for regenerative therapies for APAP‐induced ALF.

Grant Funding Source: NIH ‐ R01 DK098414, P20 RR021940‐07 and AASLD‐ALF Liver Scholar Award (Udayan Apte)
Death‐associated protein kinase mediates myofibril degeneration and myocyte apoptosis induced by beta‐adrenergic receptors (404.3)
Authors: Guo Xinji , Hooshdaran Bahman , Rafiq Khadija , Xi Hang , Kolpakov Mikhail , Houser Steven , Koch Walter , Liggett Stephen , Sabri Abdelkarim
Published: June 2021

Background: In cardiac muscle, persistent stimulation of β‐adrenergic receptors (β‐ARs) and calcium (Ca)2+ signaling plays a central role in regulating contractility, hypertrophy, and apoptosis. The Ca2+/calmodulin‐regulated Serine/Threonine kinase, death‐associated protein kinase (DAPK), regulates cytoskeletal dynamics and cell death. However, the role and activation mechanisms of DAPK in cardiac myocytes and heart failure (HF) are unknown. This study examines the role of DAPK in myocyte death induced by β‐AR stimulation.

Methods and Results: We found increased DAPK expression and activation in human failing hearts and in hearts of cardiac‐specific β1‐ARs transgenic mice. This activation of DAPK correlated with myofibril degeneration, myocyte death and cardiac contractile dysfunction, pointing to a possible role of DAPK in the development of HF. We therefore assessed whether DAPK activation was involved in myocyte apoptosis secondary to excess exposure of catecholamine. Chronic treatment with β1‐AR agonist or adenoviral expression of β1‐ARs induced DAPK activation, which was dependent on intracellular Ca(2+) elevation and calcineurin activation. Adenoviral expression of wild‐type DAPK or a constitutively active DAPK mutant significantly enhanced myofibril degeneration and myocyte apoptosis induced in response to β1‐AR stimulation. In contrast, expression of a dominant negative DAPK mutant blocked β1‐AR‐induced myocyte apoptosis and promoted myocyte hypertrophy and myofibril organization.

Conclusions: These findings reveal an unprecedented role of DAPK as a regulator of myofibril organization and myocyte apoptosis induced by persistent stimulation of β1‐AR. Thus, modulating DAPK activation to attenuate myofibril degeneration and myocyte death could be considered as a therapy target to treat patients with HF.

Grant Funding Source: NIH R01 360998
microRNA‐26b modulates NF‐κB pathway by regulating PTEN in alveolar macrophages (406.2)
Authors: Zhang Li , Huang Chaoqun , Guo Yujie , Liu Lin
Published: June 2021

NF‐κB is one of the best‐characterized transcription factors providing the link between early membrane‐proximal signaling events and changes in gene expression. It regulates the expression of many genes that play a role in inflammation. MicroRNAs are small non‐coding RNAs that can either cleave specific mRNAs or inhibit mRNA translation and subsequently silence target genes to modulate signaling pathways. Here, we report that miR‐26b, a lipopolysaccharide (LPS) ‐ responsive microRNA, repressed the phosphatase and tensin homolog (PTEN) involved in the activation of NF‐κB signaling pathway in macrophages. miR‐26b was down‐regulated in bovine alveolar macrophages after 6 and 36 h stimulation of LPS. Furthermore, we demonstrated that miR‐26b enhanced the LPS‐induced expression of tumor necrosis factor‐α (TNF‐ α), Interleukin‐1 β (IL‐1β) and Interleukin‐10 (IL‐10), but inhibited Interleukin‐6 (IL‐6). In addition, we found that miR‐26b directly bind to the 3’ untranslated region of PTEN and affected NF‐κB activity. Taken together, we conclude that miR‐26b modulates NF‐κB pathway by targeting PTEN in LPS‐stimulated bovine alveolar macrophages. These studies may provide a potential therapeutic target for bovine respiratory disease.

Grant Funding Source: supported by the Agriculture and Food Research Initiative Competitive Grant no. 09PRE2300211
Cellular mechanisms of addiction (468.2)
Authors: Kenny Paul
Published: June 2021

Smokers consume tobacco to obtain rewarding effects of nicotine, but to successfully maintain the habit they must titrate their intake to avoid noxious effects of nicotine. In contrast to nicotine reward, very little is known about brain circuitries that regulate nicotine avoidance. I will present evidence that nicotine activates neurons in the nucleus tractus solitarius (NTS) that synthesize the neuropeptide glucagon‐like peptide 1 (GLP‐1). The antidiabetic drugs sitagliptin (Januvia) and exenatide (Byetta), which inhibit GLP‐1 breakdown and stimulate GLP‐1 receptors (GLP‐1Rs), respectively, will be shown to decrease nicotine intake in mice. Chemicogenetic stimulation of GLP‐1 neurons in NTS using DREADDs also decreases nicotine intake. Conversely, GLP‐1R knockout mice are more sensitive to nicotine reward and consume greater quantities of nicotine than wildtype mice. The interpeduncular nucleus (IPN) densely expresses GLP‐1Rs and GLP‐1‐containing neurons project from NTS to IPN. Using optogenetic and electrical stimulation combined with single‐cell recordings, I will show that GLP‐1 activates excitatory inputs from medial habenula to IPN. Pharmacological stimulation of GLP‐1Rs in IPN will be shown to abolish nicotine reward and decrease intake, whereas their blockade is shown to increase intake. It is concluded that GLP‐1 neurons act as “satiety sensors” that regulate nicotine intake by controlling the activity of the habenulo‐interpeduncular aversion system.
Assembly and interrogation of regulatory networks elucidates synergistic dependencies in human malignancies (475.3)
Authors: Califano Andrea
Published: June 2021

The recent onslaught of molecular data, across multiple human malignancies, is producing an unprecedented repertoire of genetic and epigenetic alterations contributing to tumorigenesis and progression. Yet, the direct impact of this knowledge on tumor treatment and prevention is still largely unproven. Loss of tumor suppressor function is difficult to target pharmacologically and, with a handful of exceptions, alterations providing potential drug targets are relatively infrequent in cancer patients and are thus unlikely to support clinical development.

By reconstructing and interrogating the in vivo regulatory logic of distinct cancer cells, which integrates multiple aberrant signals resulting from genetic and epigenetic alterations, systems biology is starting to elucidate and mechanistically validate both oncogene and non‐oncogene addiction mechanisms. These mechanisms are exquisitely dependent on the molecular landscape of cancer subtypes, can be targeted pharmacologically, and are frequently synergistic, thus providing uniquely specific entry points for combination therapy.

In this presentation, we will discuss recent result in the discovery of synergistic, non‐oncogene addiction mechanisms and their application to the stratification and treatment of prostate cancer, high‐grade glioma, and non‐small cell lung cancer. This approach is highly extensible and has been applied to a variety of additional tumor subtypes, to the study of stem cell differentiation, reprogramming, and pluripotency control, as well as to the study of neurodegenerative diseases, such as ALS.
Dynamic control of 26S proteasome function at CNS synapses (478.3)
Authors: Patrick Gentry
Published: June 2021

The selective strengthening and weakening of synapses in response to past experience and activity is known as synaptic plasticity and is fundamental to all brain processes such as the ability to learn, form memories and modify behavior. In contrast, impairments in synaptic plasticity are considered to contribute to neuropsychiatric disorders and neurodegenerative diseases. Synaptic plasticity is largely mediated by both morphological and functional modifications to synapses. Local protein synthesis and protein degradation are known to contribute to these modifications. It is now clear that regulated proteolysis via the ubiquitin proteasome system (UPS) plays a major role in the growth and development, maintenance and remodeling of synaptic connections in the brain. We and others have previously found a novel form of regulation for the 26S proteasome in neurons involving the plasticity kinase Ca2+/calmodulin‐dependent protein kinases II alpha (CaMKIIα). CaMKIIα phosphorylates the ATPase 19S regulatory particle (RP) subunit, Rpt6, at serine 120 (S120) in an activity‐dependent fashion to modulate the activity and control the distribution of proteasomes in neurons (Djakovic et al., 2009; 2012; Bingol et al., 2010). We find that Rpt6 phosphorylation and proteasome function regulates synaptic strength and activity‐dependent generation of dendritic spines (Djakovic et al., 2012; Hamilton et al., 2012). I will discuss our more recent work and the functional relevance of proteasome phosphorylation in neurons using newly generated Rpt6 Ser120Ala and Ser120Asp (phospho‐dead and phospho‐mimetic, respectively) genetically modified mice.
Oncostatin M induces epithelial to mesenchymal transition and increased invasiveness in hepatic cancer cells through redox mechanisms (488.6)
Authors: Paternostro Claudia , Cannito Stefania , Colombatto Sebastiano , Novo Erica , Parola Maurizio
Published: June 2021

Oncostatin M (OSM), a member of IL‐6 family of cytokines, can affect hypoxia‐modulated liver processes contributing to chronic liver disease progression and hepatocellular carcinoma (HCC) development. In this study we investigated whether OSM may induce epithelial‐to‐mesenchymal transition (EMT) in human hepatic cancer cells and increased invasiveness by employing a) morphological, molecular and cell biology techniques in HepG2, HuH7 and Madin‐Darby canine kidney (MDCK) cells exposed to human recombinant OSM or b) immunohistochemistry on liver specimens from HCV cirrhotic patients carrying G1 and G2 HCC.

OSM induced EMT‐related changes (SNAI1/2 and β‐catenin nuclear translocation, E‐cadherin down‐regulation, up‐regulation of fibronectin, α‐smooth muscle actin and matrix metalloprotease‐2) and increased invasiveness in all cells. This involved early intracellular generation of reactive oxygen species (ROS), phosphorylation/inactivation of GSK‐3β and activation of PI‐3K, ERK1/2, JNK1/2, p38MAPK and STAT3. Cancer cell invasiveness was prevented by inhibiting ERK1/2, PI3K or JNK1/2 or by preventing ROS generation. Finally, OSM was expressed in HCC tumor cells in areas also positive for hypoxia‐related antigens. OSM can induce EMT in human hepatic cancer cells and stimulate invasiveness through redox‐dependent activation of EMT‐related critical kinases and transcription factors.
Molecular basis of histone acetyllysine recognition by the BRPF1 bromodomain (537.1)
Authors: Lubula Mulu
Published: June 2021

Covalent modifications on histone tails play a key role in determining the outcome of many nuclear processes including transcription, DNA repair, recombination, and replication. In humans, translocation of the monocytic leukemia zinc finger (MOZ) histone acetyltransferase (HAT) complex has been linked to a subtype of acute myeloid leukemia (AML). MOZ forms tetrameric complexes with ING5 (inhibitor of growth 5), EAF6 (Esa1‐associated factor 6 ortholog), and the bromodomain‐PHD finger protein. BRPF proteins have been shown to bridge the association of MOZ with ING5 and EAF6. Deletion mapping studies have also revealed that the acetyltransferase domain of MOZ is sufficient for the interaction with BRPF1. BRPF proteins therefore play a key role in assembling and activating MOZ acetyltransferase complexes. BRPF1 contains a unique combination of domains typically found in chromatin‐associated factors, including PHD (plant homeodomain) fingers, a bromodomain and a PWWP (Pro‐Trp‐Trp‐Pro) domain. The bromodomain and PWWP domain of BRPF1 help recruit MOZ to distinct sites of active chromatin. Bromodomains are highly conserved motifs generally known to acetylated lysines on the histone tail, but the ligands recognized by BRPF1 bromodomain are currently unknown. In this study we identified the N‐terminal histone tail ligands for the BRPF1 bromodomain, characterized the protein‐ligand binding affinities and determined the functional importance of these interactions. We also used extensive molecular dynamics simulations to generate structural models of bromodomain‐histone ligand complexes, to analyze H‐bonding and other interactions, and to calculate the binding free energies. Our results outline the molecular mechanism driving binding specificity of the BRPF1 bromodomain for discrete acetyllysine residues on the N‐terminal histone tails. A better understanding of this process will be important for the development of drugs to treat acute myeloid leukemia and other cancers.

Grant Funding Source: NIH
The role of non‐muscle myosin II during s‐looping of vertebrate heart development (540.4)
Authors: Hoang My , LaGraff Justin , Chu‐LaGraff Quynh
Published: June 2021

The early development of the heart in vertebrates consists of two phases: c‐looping and s‐looping. During cardiac looping, the relatively straight heart bends rightwards, undergoing the formation of a distinctive c‐shape and then s‐shape. Until relatively recently, it was assumed that non‐muscle II myosin is essential for heart looping. These conserved proteins interact with actin and troponin C via a Rho signaling pathway. A previous study concluded that myosin‐based contraction plays, at most, a minor role during first phase of cardiac looping, c‐looping. Here, we investigate whether non‐muscle II myosin‐based contraction was necessary for the second phase of cardiac looping, s‐looping. During heart looping, non‐muscle myosin II proteins co‐localize with actin at cell junctions. Using Verapamil, an L‐type Ca+ channel blocker, the effect of blocking actin‐myosin interactions on s‐shaped morphogenesis during chick heart development was determined. Results indicate that embryos treated with low concentrations of Verapamil exhibited heartbeat cessation, unsuccessful s‐looping, and abnormal actin distribution. Embryos treated with low concentrations of non‐muscle myosin II inhibitor Blebbistatin, and a Rho kinase inhibitor Y27632 also displayed delayed and abnormal heart looping, albeit not as severe as Verapamil‐treated embryos. We will present these studies detailing the requirement of non‐muscle myosin II during S‐looping.
Cardiac fibroblasts network organization and activation in the junctional region of the giant danio heart (543.8)
Authors: Manalo Trina , Roell Julia , Johnson Adedoyin , Lafontant Pascal
Published: June 2021

The Giant danio (GD) and zebrafish (ZF) are small fish belonging to the diverse family of cyprinids; they both display the remarkable ability for heart regeneration. Our lab has recently described the complex junctional region (JR) at the interface of the compact and spongy myocardial layers of the ZF. However, how these two distinct layers are integrated in the GD has not been studied. Using light, scanning, and transmission electron microscopy (TEM), our study reveals that the GD possesses a complex JR with transitional cardiac myocytes (CM) linking compact heart CM and CM of the spongy heart, and that luminal and abluminal contacts are mediated through discrete membrane appositions and adherens junctions. More importantly we also found a complex network of fibroblasts (FB) with extensive cytoplasmic processes occupying the JR and investing the GD compact heart. To ascertain (FB) function, we treated GD with low dose of the antineoplastic agent Doxorubicin (DOX). Using Masson’s trichrome staining, we observed the accumulation of collagen in the compact heart. In addition, TEM shows increased level of collagen in the JR of treated hearts, as well as increase in FB endoplasmic reticulum and cytoplasmic polyribosomes suggesting a state of activation. We conclude that cardiac FB are resident in the complex JR and compact heart of the GD. Moreover our data suggest they are key to the accumulation of collagen that accompany the DOX‐induced injury response.

Grant Funding Source: Supported by FDC‐DePauw University
Ischemic disease of the heart and professional adaptability of motor transport drivers (547.6)
Authors: Tursynbetov E.A. , Shaidarov M. Z. , Dubitsky A.A. , Karp L.L.
Published: June 2021

Purpose: to determine the frequency of ischemic disease of the heart at drivers of motor transport and to reveal the changes of professional functions.

Materials and methods. Cards of ambulance calls for the last 5 years are analyzed and psychophysiological testing (PPhT) of 248 drivers with a syndrome of coronary insufficiency is carried out. Starting PPhT is performed during 1‐2 days after a call of emergency doctor, final ‐ for the 6‐7 day after applying medical aid.

Results. The number of drivers’ referrals concerning to cardiovascular diseases is increased in 1,5 times. The percentage of hospitalization is increased in 12%. The particular interest is represented by results of PPhT with cardiovascular urgent conditions in dynamics. Initial testing revealed changes of latent and motor reaction, tracking speed for the moving object, testifying about the system deterioration of operative response. Final testing found positive dynamics (from 0,785±0,078 till 0,704±0,765), motor (from 1,609±1,065 till 1,509±1,120) and the tracking speed for the moving object (from 23,5±2,1 till 22,3±1,9). At all drivers system parameters of operative response authentically differed from psychophysiological indicators of healthy drivers (р<0,001).

Conclusions. PPhT authentically showed deterioration of professional qualities of drivers that increased the emergence risk of the road conflicts.
STAT3 regulation by S‐nitrosylation: implication for inflammatory disease (555.8)
Authors: Won Jeseong , Kim Jinsu , Singh Avtar , Sharma Anand , Singh Inderjit
Published: June 2021

S‐nitrosylation and S‐glutathionylation, redox based modifications of protein thiols, are recently emerging as an important signaling mechanisms. Here, we assessed S‐nitrosylation based regulation of JAK2/STAT3 pathway that plays critical roles in immune/inflammatory responses and tumorigenesis. Our studies show that STAT3 in stimulated microglia underwent two distinct redox‐dependent modifications, S‐nitrosylation and S‐glutathionylation. STAT3 S‐nitrosylation was associated with iNOS‐produced nitric oxide (NO) and S‐nitrosoglutathione (GSNO), while S‐glutathionylation of STAT3 was associated with cellular oxidative stress. NO produced by iNOS or treatment of microglia with exogenous GSNO inhibited STAT3 activation via inhibiting STAT3 phosphorylation (Tyr705). Consequently, the IL‐6 induced microglial proliferation and associated gene expressions were also reduced. In cell free kinase assay using purified JAK2 and STAT3, STAT3 phosphorylation was inhibited by its selective preincubation with GSNO, but not by preincubation of JAK2 with GSNO, indicating that GSNO‐mediated mechanisms inhibit STAT3 phosphorylation through S‐nitrosylation of STAT3 rather than JAK2. In this study, we identified that Cys259 was the target Cys residue of GSNO‐mediated S‐nitrosylation of STAT3. The replacement of Cys259 residue with Ala abolished the inhibitory role of GSNO in IL‐6‐induced STAT3 phosphorylation and transactivation, suggesting the role of Cys259 S‐nitrosylation in STAT3 phosphorylation. Our results indicate the regulation of STAT3 by NO‐based post‐translational modification (S‐nitrosylation). These findings have important implications for development of new therapeutics targeting STAT3 for treating diseases associated with inflammatory/immune responses and abnormal cell proliferation, including cancer.

Grant Funding Source: NS072511, BX001062, NS037766 and BX001072
Nucleosome binding and cell‐type specific gene expression (561.4)
Authors: Gjidoda Alison , Tagore Mohita , McAndrew Michael , Floer Monique
Published: June 2021

How nucleosome occupancy at transcriptional regulatory regions is altered to allow expression of inducible genes in mammals is not clear. We have used a quantitative assay to analyze changes in nucleosome occupancy upon induction of three pro‐inflammatory cytokines in primary mouse macrophages and find that distal and proximal enhancers are rapidly cleared of nucleosomes. We also show that the cleared enhancers become associated with cis‐regulatory transcription factors required for induction of the associated genes. Surprisingly, we find that the promoters of two of these genes are not cleared of nucleosomes under conditions that lead to high levels of gene expression, and we speculate that competing nucleosomes at the promoters may contribute to the high degree of stochasticity found at these and other inducible genes. Our studies further revealed that enhancers of these inducible genes are only lowly occupied in macrophages prior to induction, suggesting that a permissive chromatin architecture may keep enhancers accessible to subsequent binding of cis‐regulatory transcription factors. Using siRNA we have begun to dissect the involvement of lineage‐specific transcription factors (e.g. PU.1, C/EBPβ, Runx) and nucleosome remodelers (e.g. Brg1/Brm) in determining cell‐type specific chromatin architecture at enhancers of inducible genes in macrophages.

Grant Funding Source: AHA 13SDG17260004