bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of yeast deletions strains that alter the efficacy of antifungal drugs
Authors: Bataba Eudoxie , Serratore Nina D. , Briggs Scott D.
Published: June 2021

In recent years, a significant increase in mortality of patients with fungal infections has been observed. This is primarily due to a higher incidence of individuals who are immunocompromised including HIV‐infected patients, patients undergoing cancer therapy, organ transplant recipients, and patients with advanced stages of diabetes. A complication in treating patients with fungal infections is the development of drug resistant fungi. With the limited number of effective antifungal drugs and the increase of drug resistance, fungal infections and pathogenesis is a major threat to human health. The proposed studies help to solve this issue by identifying genes that alter antifungal drug sensitivity. Our central hypothesis is that a subset of transcriptional regulators are needed for pathways critical for altering antifungal drug resistance. This project used growth assays to determine the effects of the antifungal drugs, ketoconazole and amphotericin B, on yeast deletions strains. Overall, the proposed work will have an impact on human health related to fungal pathogenesis and drug resistance.

Support or Funding Information

National Science Foundation
Factors that differentiate Traumatic Brain Injury (TBI) patients from Post‐Traumatic Stress Disorder (PTSD) patients among Operation Enduring Freedom/Operation Iraqi Freedom (OEF/OIF) veterans
Authors: Miller Stacy‐Ann , Gautam Aarti , Chakraborty Nabarun , Sowe Bintu , Hoke Allison , Yang Ruoting , Hammamieh Rasha , Jett Marti
Published: June 2021

Traumatic Brain Injury (TBI) has become the “signature wound” of Operation Enduring Freedom/Operation Iraqi Freedom (OEF/OIF). Due to the increased use of improvised explosive devices by the insurgent forces and the unavoidable facial exposure of our combat troops, even when wearing protective gear, estimates suggest that as many as 20 % to 30 % of returning soldiers may eventually exhibit symptoms of TBI. Post‐Traumatic Stress Disorder (PTSD), as defined by the American Psychiatric Association, is a serious behavioral health disorder, and it has been estimated that 17.1 % of soldiers returning from Iraq, and 11.2 % of those returning from Afghanistan, have experienced major depression, generalized anxiety, and/or PTSD. As a part of the present effort, investigators within the Integrative Systems Biology Program (ISB), at US Army Center for Environmental Health Research (USACEHR), are working with collaborators from the Dwight D. Eisenhower Army Medical Center (DDEAMC) to screen for potential biomarkers associated with neuronal injury. In this pilot study, blood samples were collected at DDEAMC from patients diagnosed with PTSD, TBI, or both, and shared with USACEHR for multi‐omics analysis for identification of candidate genes/proteins distinguishing between TBI and PTSD. The candidate gene/protein approach is viewed as only a first step toward identifying molecular mechanisms that are likely to be involved in the physiologic consequences of TBI/PTSD. Gene expression, DNA methylation, and targeted proteomics analysis have been completed with these samples. Functional pathway predictions indicate possible overlaps of networks enriched by differentially expressed genes and methylated genes. The advancements realized through the current findings need to be validated using larger sample sizes, and it will be important to include a gender/age/ethnicity‐matched control. The results of this research will be leveraged in multiple ways to improve the health and treatment of returning soldiers diagnosed with TBI/PTSD.

Support or Funding Information

DISCLAIMERS: Research was conducted in compliance with all Federal Requirements. The views expressed are those of the authors and do not constitute endorsement by the U.S. Army.
Phosphatidic Acid‐Protein Phosphatase 2A Interactions Regulate Haloptropic Bending in Rice
Authors: Han Eun Hyang , Petrella Dominic , Lin Jinshan , DeLong Alison , Blakeslee Joshua J.
Published: June 2021

Rice production, which accounts for roughly 20% of the world's caloric intake, is increasingly threatened by drought or salinity stresses resulting from the depletion of freshwater reserves, the use of contaminated well water, or the aggregation of agricultural fertilizers. The root system is generally the first part of the plant to experience the osmotic and ionic trauma induced by salt stress. One method by which rice roots adapt to salt stress is through tropic growth away from regions with increased salt concentrations (i.e., halotropism). Tropic bending of roots is caused by asymmetrical re‐distribution of auxin, which results in part from stress‐induced re‐localization of PIN efflux proteins, and may also involve modulations of auxin metabolism. While halotropism has been demonstrated in several dicot species (Arabidopsis thaliana and soybean), it has not yet been examined in rice. Here we discuss the development of a multi‐tiered metabolomics platform to dissect salt stress responses (i.e. halotropic bending) in rice or other plant species. Using this platform, we have demonstrated that rice roots exhibit halotropic growth, and that this growth involves the coordinated production of phosphatidic acid, the regulated activity of the protein phosphatase 2A (PP2A) enzyme complex, and changes in the local accumulations of auxins and other phytohormones. Our data allow the generation of a model in which salt‐stress induced production of phosphatidic acid functions, at least in part, to regulate PP2A activity, modulating halotropic growth responses.

Support or Funding Information

This material is based upon work supported by the National Science Foundation under Grant No. 0446039, and USDA Grant #2007‐35304‐18418. Salaries and research support provided by state and federal funds appropriated to The Ohio State University, Ohio Agricultural Research and Development Center (SEEDS Grant 2012‐027).
A New Link Between Stress and Infection
Authors: Sule Nitesh , Pasupuleti Sasikiran , Kohli Nandita , Menon Rani , Dangott Lawrence , Manson Michael , Jayaraman Arul
Published: June 2021

We have known for a long time that stress increases the risk of bacterial infections by weakening the immune system. However, that is only one side of the story. Could bacteria be actively sensing stress to identify where and when to invade?

During conditions of stress, key markers such as the hormones norepinephrine (NE) and epinephrine are secreted into the gut by gut associated lymphoid tissue (GALT). Under normal circumstances, these tissues function as the watchtowers of the immune system. However, numerous studies have shown that GALTs are also the preferred sites for initiation of infections in the gut. We therefore hypothesized that bacteria might specifically locate GALTs by sensing the hormones as chemoattractants and this formed the basis of our studies.

We conducted chemotaxis assays with wild‐type and mutant strains of E. coli K12 (RP437) and found that NE is not a chemoattractant. Instead, we discovered that these bacteria convert NE to 3,4‐dihydroxymandelic acid (DHMA), and that DHMA is a highly potent chemoattractant. We have shown that E. coli use TynA and FeaB to convert NE to DHMA, and Tsr to sense DHMA as a chemoattractant (Pasupuleti et al. (2014) J Bacteriol. 196, 3992).

In this report, we show that DHMA is not only an attractant for enterohemorrhagic E. coli O157:H7, but it also induces virulence in this pathogen. Exposure to DHMA increased expression of LEE genes up to 35‐fold and increased attachment to HeLa cells over two‐fold. We also show that DHMA is present in the gut and that it is likely produced by gut microbiota. Using LC‐MS, DHMA could be detected and quantified in metabolite extracts from fecal samples as well as cecal contents of normal mice whereas no DHMA could be detected in fecal extracts of germfree mice.

Our studies lead us to hypothesize that normal gut microbiota convert stress hormones such as NE into DHMA. This DHMA then acts as a beacon to other bacteria, including pathogens, directing them to NE rich sites such as the GALTs, thus playing an important role in initiating infections during stress. We are now investigating the detailed molecular mechanisms involved in the activity of DHMA as a chemoattractant and a virulence factor. Our studies could help identify new strategies to fight against intestinal infections.

Support or Funding Information

National Science Foundation award MCB 1121916
Low Plasma Carotene Concentrations Are Associated with an Increased Risk of Acute Coronary Syndrome in a Korean Population
Authors: So Jisun , Walker Maura E , Matthan Nirupa R , Lichtenstein Alice H , Han Sung Nim , Chung Ick‐Mo , Johnson Elizabeth J
Published: June 2021

Acute coronary syndrome (ACS) includes acute myocardial infarction and unstable angina, which are manifestations of coronary atherosclerosis. Circulating concentrations of carotenoids have been suggested to exert a protective effect in the development of coronary heart disease. We investigated whether plasma carotenoids – xanthophylls (lutein, zeaxanthin, and beta‐cryptoxanthin) and carotenes (alpha‐carotene, beta‐carotene, and lycopene) – were related to the risk of developing ACS in middle‐aged Korean adults. Included were 30 subjects diagnosed with ACS (cases) and 30 age‐ and sex‐matched control subjects. Plasma concentrations of carotenoids were measured by high performance liquid chromatography. The odds ratio (OR) of each plasma carotenoid was estimated by a logistic regression model adjusted for waist circumference.

Compared to the controls, the cases had a significantly greater waist circumference (85.0 ± 8.6 cm vs. 80.6 ± 8.3 cm; p < 0.05) and lower HDL cholesterol concentration (45.9 ± 11.9 vs. 51.6 ± 11.7 mg/dl; p < 0.05). Plasma concentrations of carotenes, but not xanthophylls, were significantly lower in the cases as compared with controls (alpha‐carotene: 0.11 ± 0.05 vs. 0.17 ± 0.18 umol/l, p < 0.05; beta‐carotene: 1.00 ± 0.62 vs. 1.54 ± 0.88 umol/l, p < 0.05; lycopene: 0.21 ± 0.16 vs. 0.62 ± 0.39 umol/l, p < 0.001). After adjusting for waist circumference, plasma concentrations of beta‐carotene (OR = 0.92, 95% CI 0.85–0.99; p = 0.025) and lycopene (OR = 0.57, 95% CI 0.42–0.79; p < 0.001) were associated with lower risk of ACS. No significant association with the risk of ACS was observed for plasma xanthophyll concentrations.

This study provides evidence for inverse associations between plasma beta‐carotene and lycopene and the risk of developing ACS, suggesting that consumption of fruits and vegetables (especially, carrots, winter squash, tomatoes) may play a protective role in the development of ACS in the Korean population.

Support or Funding Information

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF‐2012R1A1A2008597) and USDA 1950‐51000‐087.
A multi‐level risk factor assessment for stunting: Evidence from a national sample in Nepal
Authors: Dorsey Jamie , Manohar Swetha , Neupane Sumanta , Shrestha Binod , Thorne‐Lyman Andrew , Webb Patrick , Klemm Rolf DW , West Keith P
Published: June 2021

BACKGROUND

Despite marked improvement in child undernutrition, especially stunting, in Nepal over the past decade, stunting prevalence remains high at 41% thus generating an urgent need to understand factors associated with childhood stunting.

OBJECTIVE

Identify individual, household and community level factors associated with stunting in Nepali children 6–59 months of age.

METHODS

The Policy and Science for Health, Agriculture, and Nutrition (PoSHAN) Community Studies has established a national agriculture‐to‐nutrition surveillance system in a representative sample of ~5500 households in 63 wards across 21 districts, 7 in each of the 3 agro‐ecological zones of the country. This study presents cross‐ sectional data collected in 2013 among a sample of 4853 children to determine factors associated with under‐five stunting, indicated by a height‐for‐age (HAZ) < −2. Using mixed effects logistic regressions to estimate adjusted odds ratios (AOR) with 95% confidence intervals (CI), we examined individual, household and community level risk factors associated with stunting, adjusting for the multi‐level clustering design.

RESULTS

The national prevalence of stunting among children < 5 years was 37.5%. Children living in less developed communities had a higher odds of stunting than those who lived in more developed communities [AOR (95% CI): 2.38 (1.36, 4.14)]. Female household headship was associated with lower odds of stunting [0.78 (0.64, 0.95)]. The presence of 3 or more under‐five year old children in a household was associated with an increased risk of stunting risk [1.19 (1.44, 2.52)] versus households with only one <5 year old. Monthly expenditure (below the 20th vs above the 80th%ile of household expenditure) was associated with an increased stunting risk [1.68 (1.27, 2.24)]. At the individual level, wasting of the child [1.52 (1.24, 1.86)], high fever in the last 30 days [1.28 (1.03, 1.59)], and maternal factors such as a lack of formal education [2.09 (1.48, 296)] and short stature [2.52 (1.96, 3.25)] were associated with an increased risk of stunting.

CONCLUSION

This study identifies risk factors at community, household and individual maternal and child levels associated with preschool child stunting that may assist in targeting programs to highest risk groups in the future.

Support or Funding Information

USAID, Feed the Future Lab for Nutrition
Igniting Youth to Create Healthier Communities: Year 4 Policy Changes
Authors: Kattelmann Kendra , Chen Biyi , Erickson Megan , Zies Susan , Jess Marcia , Li Yijing , Muturi Nancy , Lindshield Erika , Remley Daniel , Kidd Tandalayo
Published: June 2021

Objective

The growing rate of childhood overweight and obesity suggests a need for more effective obesity prevention programs. Community‐Based Participatory Research (CBPR) is a collaborative approach to research that includes equal contributions from community members, organizational representatives and researchers in all facets of the research process. The approach allows participants to share their expertise in order to enhance knowledge and develop interventions that benefit the community. Ignite is five‐year, multiple‐state (Ohio, South Dakota and Kansas) program using the CBPR process to address overweight and obesity among 6–8th grade youth. As part of the Ignite program, virtual dialogues were held with key stakeholders (community leaders, school administrators, and adolescents) who were actively involved in the CBPR project to identify policy and program changes attained during Ignite implementation.

Study Design, Setting, and Participants

At the end of year four of programming, a virtual forum with steering committee members (7 school administrators and staff, 2 community members, 10 youth) and 11 researchers from the three intervention communities was conducted. Participants identified priority issues and program implementation associated with Ignite.

Outcome, Measures and Analysis

Virtual forum discussion was recorded, transcribed, and thematically analyzed using NVivo software (QSR International Pty Ltd. Version 10, 2012).

Results

Policy and program changes that support healthful eating behaviors and physical activity (PA) were identified. The school food and nutrition policy changes included enhancing availability of breakfast and healthful snacks and adding nutrition education to physical education (PE) class. The PA policy changes included increasing the number of programs to support walking and more physical activities. In addition, policy changes related to both nutrition and PA included incorporating wellness into curriculum, devoting time and space to wellness, and exploring funds for sustainability of programing beyond the grant.

Conclusions and Implications

CBPR programming promoted positive policy and program changes related to healthier dietary and physical activity environment in the intervention schools and communities.

Support or Funding Information

This material is based upon work that is supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture, under award number 2012‐68001‐19619
Effectiveness of the Children's Healthy Living (CHL) Multilevel Multicomponent Community Intervention Program in 5 US affiliated Pacific Jurisdictions
Authors: Novotny Rachel , Wilkens Lynne R , Nigg Claudio R , Braun Kathryn , Butel Jean , Areta Aufai , Coleman Patricia , Belyeu‐Camacho Tayna , Greenberg Joshua , Bersamin Andrea , Guerrero Rachael Leon , Barber Leroy Robert , Fialkowski Marie Kainoa , De la Cruz‐Talbert Elise
Published: June 2021

The Children's Healthy Living (CHL) program, a collaboration among Pacific Land Grant Colleges, was developed and implemented with 27 communities across the US affiliated Pacific region in a 24 month community randomized intervention trial to prevent young child obesity. The multilevel, multisite, community‐based intervention consisted of 4 cross‐cutting functions (initiate or strengthen school wellness policies; partner and advocate for environmental change; promote CHL messages; and local capacity building) with 20 activities addressing 6 targeted behaviors (physical activity, sedentary behavior, sleep, fruit and vegetable intake, sugar sweetened beverage intake, and water intake). A common intervention template and outcome measures were developed and implemented; IRB approval or ceding of approval was obtained at each of the 5 collaborating institutions. Controlling for age and sex of 2–10 year old children and accounting for the randomization units and clustering, the CHL intervention communities had significant changes in the following variables compared to control communities: prevalence of acanthosis nigricans (change of −0.1% vs +0.2%), child overweight plus obesity (−3.0% vs +1.7%), child waist circumference (+0.2cm vs +9cm), and daily screen time (−2.4min vs +3.0mins). Accelerometry and food related variables (fruit and vegetable intake, sugar sweetened beverage intake, water intake) are still being analyzed. The results indicate that community‐based multilevel multi‐component interventions can impact young child overweight and obesity and related behaviors and conditions.

Support or Funding Information

USDA/NIFA grant 2011‐68001‐30335
Screen Time is Associated with Lower Healthy Eating Index (HEI)‐2010 Total Score in Low‐Income Hispanic Pregnant Women in the Starting Early Study
Authors: Berube Lauren Thomas , Woolf Kathleen , Gross Rachel , Messito Mary Jo
Published: June 2021

Background

Sedentary behaviors, including screen time (i.e., time spent using televisions, computers, and smart phones), are associated with unhealthy dietary behaviors and obesity. However, it remains unknown how screen time and dietary habits are related during pregnancy, a critical period known to impact both maternal and child health outcomes.

Objective

To examine dietary quality and screen time in low‐income Hispanic pregnant women, known to be at high risk of obesity.

Methods

Cross‐sectional analysis of baseline data from the Starting Early study, a randomized controlled trial beginning in third trimester of pregnancy to prevent childhood obesity. Independent variable: screen time (ST) categorized as low (≤2 hr/d), moderate (>2 to ≤4 hr/d), or high (>4 hr/d). Dependent variable: diet quality assessed using the Block 2005 Bilingual Food Frequency Questionnaire, and the Healthy Eating Index (HEI)‐2010. One‐way analysis of variance (ANOVA) and post‐hoc analyses were used to determine associations between screen time and HEI‐2010 total score.

Results

Women (N=482) did not differ in mean body mass index, but women with low and moderate ST were older than those with high ST (p<0.001). The mean HEI‐2010 total score was 72.3 ± 7.9 for low ST, 69.0 ± 9.3 for moderate ST, and 66.2 ± 10.1 for high ST (p<0.001). Low ST had higher mean empty calorie (p<0.001) and vegetable (p<0.001) scores than moderate and high ST. Low and moderate ST had lower mean daily energy intake (p=0.004) and higher mean whole fruit (p=0.003), greens and beans (p<0.001), and dairy (p=0.002) scores than high ST.

Conclusion

Less screen time was associated with better diet quality and lower intake of empty calories. High screen time was associated with greater energy intake and lower intake of foods recommended by the Dietary Guidelines for Americans, 2010. Interventions are needed to reduce screen time and increase intake of nutrient‐dense foods among low‐income Hispanic women during pregnancy.

Support or Funding Information

Funding Disclosure: USDA Grant # 2011‐68001‐30207
Associations Between Neck Circumferences and Measures of Health in Young Adults
Authors: Famodu Oluremi A , Barr Makenzie L , Colby Sarah E , Zhou Wenjun , Bryd‐Bredbenner Carol , Mathews Anne E. , Olfert Melissa Diann
Published: June 2021

Background

Neck circumference is a novel method for estimating upper‐body subcutaneous adipose tissue distribution. Studies have concluded that neck circumference is a potentially useful, simple, non‐invasive, cost‐effective initial screening tool for overweight/obesity and the metabolic syndrome.

Objective

The objective of this study was to determine the relationship between neck circumference and other anthropometric measures in young adults.

Methods

Over 1500 students across 8 Universities in the United States underwent a physical assessment. Neck circumference, height, weight, and blood pressure were taken in duplicate by trained researchers using standard procedures. Body mass index (BMI) was calculated using height and weight measures. Spearman correlations were calculated to determine associations between neck circumference and other continuous variables of health.

Results

Most study participants were Caucasian (67%), female (70%), and reported growing up in the non‐Appalachian region (82%) of the United States. The percentage of those with a BMI ≥ 25 kg/m2 was 34% and 41% for females and males, respectively. The percentage of those with a neck circumference indicating obesity (i.e. ≥ 35.5 cm for males and ≥ 32 cm for females) was 79% vs 50% for women. In both genders, significant positive correlations were seen between neck circumference and body weight (women, ρ=0.707, p<0.0001; men ρ=0.693, p<0.0001), BMI (women, ρ=0.692, p<0.0001; men ρ=0.704, p<0.0001), waist circumference (women, ρ=0.721, p<0.0001; men ρ=0.687, p<0.0001), hip circumference (women, ρ=0.692, p<0.0001; men ρ=0.638, p<0.0001), and systolic blood pressure (women, ρ=0.309, p<0.0001; men ρ=0.301, p<0.0001).

Conclusion

Overall, this study suggests that neck circumference closely correlates with other measurements related to disease, especially upper body fat. Therefore, incorporating neck circumference measurements in research is a viable, cost‐effective method for corroborating other health measures associated with obesity and its co‐morbidities.

Support or Funding Information

Approval to use the data set was granted by the University of Tennessee Institutional Review Board prior to study implementation. This material is based upon work that is supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture, under award number 2014‐67001‐21851.
Absorption and Metabolism of Isothiocyanates Formed from Broccoli Glucosinolates: Effects of Daily Consumption
Authors: Charron Craig , Vinyard Bryan , Ross Sharon , Seifried Harold , Jeffery Elizabeth , Novotny Janet
Published: June 2021

Sulforaphane is derived from glucoraphanin, the predominant glucosinolate in broccoli, and is associated with anti‐cancer effects. A preclinical study suggested that daily consumption of broccoli may increase the production of sulforaphane and sulforaphane metabolites available for absorption. To test this result in humans, we designed a randomized crossover study (n=18) during which subjects consumed a Brassica‐free control diet for 16 days or the same diet with 200 g of broccoli and 20 g of fresh daikon radish for 16 days. On day 17, all subjects consumed 200 g of broccoli with 20 g of daikon radish. Plasma and urine were collected over 24 h and analyzed for sulforaphane and metabolites of sulforaphane and erucin by tandem mass spectrometry. Females and subjects with the GSTM1+ genotype had higher concentrations of total metabolites in plasma when they had been consuming the control diet prior to day 17. Subjects with BMI>26 (median) had higher total plasma metabolites following the control diet compared to the daily broccoli diet, whereas subjects with BMI<26 had higher levels following the broccoli diet compared to the control diet. The urinary excretion rate of total metabolites was higher following the control diet compared to the broccoli diet for subjects with BMI>26. These results indicate that multiple factors affect the plasma concentrations of compounds believed to confer protection against cancer.

Support or Funding Information

Supported by the National Cancer Institute and the U.S. Department of Agriculture.
In silico screening and usability of molecular modeling in finding bio‐active compounds in diet components; investigation of javamide‐I/‐II/‐III and chlorogenic acids found in coffee and flavonoids as p38 MAP kinase inhibitors.
Authors: Park Jae
Published: June 2021

Javamide‐I/‐II/‐III are bio‐active compounds found in coffee. Our preliminary study suggested that javamide‐I/‐II may inhibit p38 MAP kinase, thereby potentially beneficial in some human diseases related to subclinical inflammation. However, potential effects of these javamides on the p38 inhibition have not been fully investigated using in silico methods. Therefore, in this paper, javamide‐I/‐II/‐III were screened related to p38 MAP kinase inhibition with other phytochemicals such as chlorogenic acids and flavonoids using in silico methods which is widely used to find and develop new effective and selective compounds. After the screening was completed with ΔE values, actual effects of the javamides on p38 MAP kinase were also determined using an in vitro assay method. Javamide‐I/‐II/‐III with best scores were able to inhibit p38 MAP kinase (IC50 of 8–36 uM) better than chlorogenic acids and flavonoids. Especially, javamide‐II found in coffee exhibited competitive inhibition of p38 MAP kinase with Ki ≈ 9.6 uM, confirming that virtual screening methods are also useful in finding bio‐active compounds with specific activities. This study suggests that combining in silico screening and in vitro assay could be a very convenient and effective method even in finding bio‐active compounds present in human diet components.

Support or Funding Information

USDA (1235‐51000‐054‐00D)
Identification of metabolite markers of long‐term green tea polyphenol intake in humans
Authors: Zhou Yuyin , Zhang Ningning , Arikawa Andrea , Chen Chi
Published: June 2021

Green tea polyphenols (GTP) possess diverse bioactivities on cell proliferation and survival, but the metabolic impact of chronic GTP intake on humans was not well defined. In this study, fecal and urine samples from postmenopausal female subjects taking the GTP supplement or placebo for 12 months were analyzed by liquid chromatography‐mass spectrometry‐based metabolomics analysis. The metabolites associated with GTP treatment were first identified by multivariate modeling of control and treatment, and then characterized by structural elucidation and quantitative analysis. A group of phenolic acids derived from microbial metabolism of GTP were identified as the robust exposure markers. In contract, major GTP, such as epigallocatechin gallate (EGCG), were not found in significant amounts in feces, suggesting extensive biotransformation mediated by microflora. One of these microbial metabolites was determined as 3‐hydroxyphenylvaleric acid, which has not been identified as an EGCG metabolite in human feces previously. In urine samples, chronic GTP treatment led to the presence of sulfate and glucuronide metabolites of GTP, but decreased the levels of microbial metabolites phenylacetylglutamine and indoxyl sulfate. Interestingly, ascorbic acid was identified as one of urinary metabolites that were significantly decreased by GTP, indicating the influence of chronic GTP treatment on redox balance. Overall, these results suggest that specific microbial, xenobiotic, and endogenous metabolic events contribute to the formation of diverse markers of long‐term GTP intake.

Support or Funding Information

NCI grant R01 CA127236

NIFA project MIN‐18‐092
Epigenetic Regulation of WNT and Hedgehog Oncogenic Signaling in Breast Cancer Cells in Response to Dietary Polyphenols
Authors: Stefanska Barbara , Kurzava Lucinda , Lubecka Katarzyna , Beetch Megan , Flower Kirsty , Flanagan James M
Published: June 2021

The Wnt and Hedgehog signaling pathways play roles in embryonic and stem cell developmental processes but can become overactive in human cancers, contributing to cancer cell invasion and metastasis. Crosstalk occurs between these two pathways in cancer due to the ability of Hedgehog signaling proteins to regulate activity of the Wnt pathway, and vice versa. This interdependent regulation allows for cancer therapies to mutually target the activity of both pathways. Interestingly, certain dietary polyphenolic compounds suppress the Hedgehog and Wnt signals in cancer. In our study, polyphenols were shown to modify epigenetic marks in genes positively regulating these pathways including SHH, a Hedgehog signaling protein, GLI2, a transcriptional activator of Hedgehog signaling, and four Wnt signaling ligands including WNT4, WNT6, WNT9A, and WNT10B.

Using the polyphenol resveratrol from grapes, we examine the role of DNA methylation in regulation of Hedgehog and Wnt signaling in breast cancer. Non‐invasive MCF‐7 and invasive MCF10CA1a human breast cancer cell lines were used as an experimental model. Following genome‐wide DNA methylation analysis with Illumina 450K array, pyrosequencing and QPCR were performed to assess methylation and expression of the selected positive regulators and downstream targets of Hedgehog and Wnt signaling.

We found 6,347 CpG sites differentially methylated upon 9‐day treatment of MCF10CA1a breast cancer cells with 15μM resveratrol compared to untreated cells. A majority of those CpG sites were hypermethylated and located within genes functionally associated with oncogenic signaling pathways, including Hedgehog and Wnt. Resveratrol led to increased methylation of enhancer regions of SHH, GLI2, and WNT4, and CpG island regions of WNT6, WNT9A, and WNT10B. Increase in methylation of these genes was confirmed by pyrosequencing in breast cancer cells with high and low invasive potential. Along with increased methylation of these regions, the expression of these genes decreased following resveratrol treatment. This was associated with downregulation of Hedgehog target gene Bcl‐2 and Wnt target gene EpCam, as well as a mutual target FOXM1, in invasive MCF10CA1a breast cancer cells.

Our results provide insight into regulation of oncogenic signals in cancer and support for epigenetic‐targeting strategies as an effective anti‐cancer approach.

Support or Funding Information

This study was supported by the Purdue University Center for Cancer Research, Indiana CTSI (UL1TR001108), Women's Global Health Institute, and USDA National Institute of Food and Agriculture (Hatch project 1005656) granted to BS.
Mixed Dishes are a Top Contributor to Vitamin K Intake in US Adults: Data from the 2011–2012 NHANES
Authors: Harshman Stephanie Geris , Finnan Emily G , Barger Kathryn , Bailey Regan L , Haytowitz David B , Booth Sarah L
Published: June 2021

The primary dietary form of vitamin K in US diets is phylloquinone (PK), which is found in green vegetables and vegetable oils. PK content of foods are provided in the USDA National Nutrient Database for Standard Reference, which helps to estimate current dietary intakes. As our food supply diversifies and expands, food groups contributing to dietary PK intake are also expanding. It is important to identify the contributors to dietary vitamin K estimates to guide healthcare professionals and recommendations on food sources of vitamin K.

The purpose of this study was to estimate: (1) the PK consumed in the diet of US adults; (2) the contribution of different food groups to PK intake in individuals with high or low vegetable intake (≥ 2 cups < 2 cups vegetables/day); and (3) to characterize the contribution of different mixed dishes to PK intake.

We used data from adults (≥ 20 y; 2092 men and 2214 women) participating in NHANES 2011–2012, a nationally‐representative complex survey, to determine usual PK intake correcting for measurement error with the National Cancer Institute method. Food group classifications were determined according to the USDA What We Eat in America food groupings. The proportion of PK supplied by each food group and mixed dishes was compared between low and high vegetable consumers, using multiple t‐tests with Bonferroni‐Dunn correction for multiple comparisons.

On average, 43.0% of men and 62.5% of women met the AI of 120 and 90 μg/day of PK, respectively. Vegetables were the highest contributor to PK intake, contributing 60.0% and 36.1% in high and low vegetable intake groups, respectively. Mixed dishes were the second highest contributor, contributing approximately 16% and 28% of total PK intake in high and low vegetable intake groups, respectively (all p<0.001 between vegetable intake groups). Within mixed dishes, grain based dishes including macaroni and cheese, pasta, and rice dishes contributed to 25% of dietary PK, followed by sandwiches (9.7%), and pizza (10%) in the low vegetable intake group. In individuals with high vegetable intake, primary sources of dietary PK in mixed dishes were soups (23%), “Asian” dishes (e.g., fried rice with soy sauce, 21.5%), and meat and poultry (17.9%).

Our findings suggest mixed dishes are a significant contributor to PK intake in the United States. Unique to mixed dishes is the large contribution of grain‐based foods that often lack PK‐rich vegetables. Additional research is required to further quantitate other vitamin K forms in the US food supply, and to identify all dietary sources of vitamin K to better guide recommendations for vitamin K intakes in the U.S.

Support or Funding Information

Supported by USDA Agricultural Research Service under Cooperative Agreement No. 58‐1950‐7‐707 and 58‐1235‐1‐193.
Dietary Fiber's Contribution to the Energy Needs of the Microbiota
Authors: Gordon Dennis Theodore , Baer David J , Mai Volker
Published: June 2021

The objective of this research is to address the amount of energy resulting from the total fermentation of an oligosaccharide, resistant maltodextrin (RM), used for the growth of the microbiota (MB). The metabolizable energy (ME) of RM has been previously reported (J. Nutr. 144:1, 2014). Dietary Fiber (DF), intrinsic and intact in plant foods, and Added Fiber, isolated or modified from plant foods or chemically synthesized are now defined (FDA, 2014) as having “beneficial physiological effects for human health (BPEFHH)”. The microbiota (MB) is considered to have strong associations with BPEFHH, but evidence to confirm this association has not been established. While DF can contribute to health benefits, its quintessential properties are to promote laxation and serve as the primary energy source for the MB. However, the energy contribution of DF for MB growth is not known. A randomized placebo controlled double blind crossover study was conducted and all participants were provided all foods for replicated intakes and fed to maintain body weight. As part of the controlled diet, men (n=14) were fed, in in random order, sequence 3‐treatments, of 0 g RM and 50 g digestible dextrin (DD), 25 g RM and 25 g DD, and 50 g RM and 0 g DD. In addition to a MB analysis of feces, total MB counts were accomplished by appropriate dilution of wet fecal samples and MB stained for FISH (counts) cell counting with a light microscope. Total wet and dry fecal weights (g), increased with 25g RM (148,32‐NS) and 50g RM (161,39‐P ≤0.02) compared to the control treatment, 0g RM (118,27), respectively. Total MB counts in wet fecal samples, in counts per g for 0, 25 and 50 g RM/day were 8.6a, 10.1a,b and 10.3b × 109, and for the total feces, total MB counts would be 1.0, 1.5 and 1.2 × 1012, respectively. The ME values for RM fed 25g and 50g/d were 8.2 kJ and 10.4 kJ levels, and net energy (NE) values were 8.2 kJ and 2.0 kJ/g, respectively. It is suggested that totally fermentable DF even in diets currently providing 18–20 g total DF/d, provide approximately 5 to 7 kJ/g for MB growth. Increased total MB in wet fecal weight, increased mass, and with increased MB concentration per g feces is considered as contributing to DF ability to improve BPEFHH, and initially via better laxation.

Support or Funding Information

Support, Matsutani, Co, LTD, USDA, ARS
Validation of Ru(II)‐caged Abiraterone as a Chemical Tool for Controlling CYP17A1 Activity with Visible Light
Authors: Li Ao , Yadav Rahul , White Jessica K. , Herroon Mackenzie K. , Podgorski Izabela , Turro Claudia , Scott Emily E. , Kodanko Jeremy J.
Published: June 2021

Cytochromes P450 (CYP) enzymes are ubiquitous heme‐containing proteins catalyzing numerous and varied biosynthetic and detoxification transformations in biological systems. Potent and selective inhibitors have been aggressively pursued as chemical tools, drugs, and insecticides, but lack spatial and temporal control over CYP inhibition in vitro and in vivo. Metal‐caged versions of such inhibitors with photoactivated release can allow researchers to interrogate biological systems spatiotemporally. Here, a novel Ru(II)‐caged complex was designed and synthesized with the CYP17A1 inhibitor abiraterone, a prostate cancer drug. While the intact caged complex is non‐toxic and exceptionally stable in the dark, upon irradiation with low energy visible light abiraterone is rapidly released. In the dark the Ru(II) caged abiraterone does not bind to CYP17A1, but light exposure results in the released abiraterone binding to CYP17A1. This binding is observed as a type II shift consistent with the inhibitor nitrogen coordinating to the CYP17A1 heme iron with a high affinity, similar to abiraterone itself. Additionally, dark Ru(II)‐abiraterone is nontoxic to DU145 human prostate cancer cells, but light‐exposed Ru(II)‐abiraterone and abiraterone resulted in similar toxicity. Overall, this demonstrates that metal‐caged inhibitors can be designed with ideal properties as potent and selective chemical tools controlled by visible light in a biological setting.

Support or Funding Information

NIH (EB 016072) and NSF (CHE1212281)
The Sphingosine‐1‐Phosphate Analog, FTY720, Reverses Diastolic Dysfunction and Hypertrophy in Familial Hypertrophic Cardiomyopathy
Authors: Ryba David M , Warren Chad M , Karam Chehade N , Davis Robert T. , Chowdhury Shamim A. K. , Alvarez Manuel G. , Wieczorek David F , Solaro R. John , Wolska Beata M.
Published: June 2021

Background

Hypertrophic cardiomyopathy (HCM) patients present with progressively worsening hypertrophy, diastolic dysfunction and fibrosis, ending in heart failure with preserved ejection fraction. At the molecular level, a common pathology found is an increase in myofilament‐calcium (Ca) sensitivity. Previously, we have shown that oxidative modification of myofilament proteins, as a result of increased oxidative stress in HCM, may be a direct mediator of this increase. FTY720 is an immunomodulating drug, clinically used in the treatment of multiple sclerosis, which we have previously identified to have anti‐fibrotic and anti‐hypertrophic effects in a model of pressure‐overload‐induced hypertrophy. We sought to understand whether agonism of the sphingosine‐1‐phosphate receptor by FTY720 would be of therapeutic benefit in HCM, and its effects on oxidative modification.

Methods

We treated an HCM mouse model linked to a mutation in tropomyosin (Tm‐E180G) and non‐transgenic (NTG) littermates with FTY720 or vehicle for six weeks. We assessed cardiac function and morphology using echocardiography and pressure‐volume (PV) relations and the myofilament‐Ca‐response using detergent‐treated (skinned) fibers. We also investigated if signaling pathways were altered.

Results

We found that FTY720‐treated Tm‐E180G mice had a significant reduction in left atrial size, E/A and E/Em ratios compared to significantly increased baseline measurements as assessed by echocardiography. PV relations revealed significant improvements in the end‐diastolic pressure volume relationship, preload recruitable stroke work, and relaxation constant (Tau). FTY720‐treated NTG mice displayed a significant decrease in ejection fraction and rate of maximum pressure development in early systole compared to vehicle‐treated NTG mice assessed by PV relations. Skinned fiber bundles revealed a significant decrease in myofilament‐Ca‐responsiveness. We attributed these functional improvements to a down‐regulation of S‐glutathionylation of cardiac myosin binding protein‐C in FTY720‐treated Tm‐E180G mice. The increases in oxidative modification in vehicle treated Tm‐E180G mice were due to an up‐regulation of NADPH oxidase 2 enzyme expression (NOX2), and fibrosis and hypertrophy to increased periostin, TGF‐beta expression, and GATA4 phosphorylation. These maladaptive changes were reversed by FTY720 treatment.

Conclusions

In Tm‐E180G mice, FTY720 improves diastolic function and morphology by reversing oxidative modification of myofilament proteins, resulting in a decrease in myofilament‐Ca‐responsiveness, via down‐regulation of NOX2 and other maladaptive remodeling signals.

Support or Funding Information

R01 HL128468 (BMW, RJS) and F31 HL127996 (DMR)
Single Amino Acid Substitutions Affect the Stability of the Dimer Interface of the E. coli β Clamp
Authors: Baez Joe
Published: June 2021

The β sliding clamp is an E. coli DNA replication cofactor and a subunit of the DNA polymerase holoenzyme responsible for tethering the DNA polymerase to the DNA primer template to prevent polymerase dissociation, increase replication processivity, and mediate polymerase switching to enhance DNA damage tolerance. The circular protein is a three‐domain homodimer arranged in a head‐to‐tail orientation with an ability to exist in an open or closed conformation at the dimer interface found between domains I and III of each subunit. The clamp's ability to open and close controls associations with DNA and dictates the efficiency of bacterial replication. In this study we introduced mutations proposed to stabilize or destabilize the dimer interface of the clamp in order to probe the mechanism of clamp opening, and find potential target residues for improving or inhibiting clamp function and DNA replication. The ability to support replication in vivo, thermostability, clamp opening function, and effect on polymerase processivity have been studied for each mutant clamp. The results support the finding that specific mutations to the L82 residue found at the β clamp's dimer interface severely destabilize the clamp and diminish E. coli's ability to replicate.

Support or Funding Information

Supported by NSF REU CHE‐1262734.
Llama Hemoglobin Binding to Heterologous Haptoglobins
Authors: Vanderpuye Oluseyi Adewale , Troutman Kicheryltus , Kellam Netanya , Dunn Christina
Published: June 2021

The plasma protein haptoglobin binds hemoglobin released from lysed erythrocytes. The complex is internalized through the macrophage membrane protein CD163 thus preventing pathological oxidation of biological molecules and tissues. Hemoglobin primary structures can be as little as 50% homologous among species. Haptoglobin also shows structural variation among species and a genetic polymorphism is found in humans and not in other mammals. The goal of this study was to examine the extent to which hemoglobins can bind to haptoglobins from different species. The llama is adapted to high altitude and its hemoglobin might thus differ from those of many other mammals. By using non‐denaturing agarose gel electrophoresis, the charge properties and binding to human haptoglobin was compared for llama, human, dog, pig, horse and goat hemoglobins.

Llama hemoglobin was found to be more positively charged than the other mammalian hemoglobins. All hemoglobins tested showed a band of increased anodal mobility when incubated with human plasma which suggested formation of hemoglobin‐haptoglobin complexes. For one plasma sample there was only one new band whereas for a second sample two new anodal hemoglobin bands were formed. This suggested the ability of the different species hemoglobins to bind to different genetic polymorphisms of human haptoglobin. When the different hemoglobins were incubated with purified haptoglobin variants Hp 1:1 and Hp 2:2, one new anodal hemoglobin‐containing band was seen. The new band in the Hp 1:1 samples had higher mobility than those in the Hp 2:2 samples. In all cases the llama hemoglobin‐haptoglobin samples had lesser mobility than those of the other mammals. Because incubation with one of the plasma samples produced two new bands rather than one as with the Hp 1:1 and Hp2:2 it is surmised that this plasma sample had the Hp 2:1 variant and that all animal hemoglobins tested can bind the three genetic variants of human haptoglobin although this variation is regarded to be absent in non‐human species. It is concluded that the binding sites on hemoglobin and haptoglobin for each other have been evolutionarily conserved despite differences in primary structure and marked difference in the charge of llama hemoglobin.

Support or Funding Information

The research was supported by Furman University Research Experience for Undergraduates 2016 session, NSF grant NSF CHE‐1460806 – awarded to K. Buchmueller and T. Hanks at Furman University and by Albany State University, Center for Undergraduate Research
Developing Monovalent Ion Parameters for the Optimal Point Charge (OPC) Water Model
Authors: Clark Daniel E , Dood John C , Krueger Brent P
Published: June 2021

Molecular dynamics (MD) simulations are used to model the structure and movement of macromolecules. The gold standard for MD is to explicitly include water molecules using one of several standard models. Recently, a new water model, Optimal Point Charge (OPC), has been developed with simulation performance that compares better to experiment than existing models in its class (Izadi, Anandakrishnan, Onufriev, J. Phys. Chem. Lett., 2014, 5, 3863‐3871). For this new water model to be useful, Lennard‐Jones (LJ) parameters must be developed for at least a few monovalent ions. In this study MD simulations were used to develop these parameters. Preliminary results are presented including: extensive convergence testing of Hydration Free Energy, Lattice Constants (LC), and first peak position of radial distribution functions (RDF's); as well as surfaces showing the dependence of the RDF and LC on various LJ parameters within the OPC water model.

Support or Funding Information

National Science Foundation awards 1058981 and 1039925.