bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel light, oxygen, or voltage fluorescent tag for structural and biophysical analysis of DNA repair enzymes (989.1)
Authors: Robertson Molly , Hatherill Robert , Fuss Jill , Tainer John 
Published: June 2021

DNA is constantly threatened by internal and external pressures. Xeroderma Pigmentosum (XP) complementation groups D (XPD) and B (XPB) are helicases that unwind helix‐distorting lesions in damaged DNA allowing for repair to occur. These proteins are critical components of the Transcription Factor II H complex, and are essential for nucleotide excision repair. Three rare genetic diseases characterized by cancer and premature aging are caused by mutations in the genes that code for these helicase enzymes. In order to understand how proteins function at the molecular level, it is necessary to investigate the 3‐dimensional structure. To accomplish this, we amplified, cloned, and expressed human XPD and XPB, as well as archeal XPD from Sulfolobus acidocaldarius (SaXPD), and an associated DNA repair protein. Since crystallographic structures will be formed in an anaerobic environment, we employed the flavoprotein improved light, oxygen, or voltage sensing (iLOV) fluorescent tag to investigate target molecules in E. coli. We purified single‐domain llama nanobodies using affinity chromatography. The anti‐iLOV variants were used to optimize large‐scale purification of the DNA repair proteins of interest. We conclude that the novel DNA iLOV fusions will facilitate anaerobic crystallography techniques.Grant Funding Source: NSF ATE DUE 1205059
Household food insecurity is associated with depression among young Cambodian women in Massachusetts (LB476)
Authors: Chen Shanshan , Peterman Jerusha , Purdue‐Smithe Alexandra , Otis Nicholas , Cordeiro Lorraine 
Published: June 2021

Background: The Cambodian refugee population in the United States is at increased risk for household food insecurity and adverse mental health. There is limited literature regarding the association between food insecurity and mental health in young Cambodian women.Objectives: The objective of this study was to investigate the association between depression status and household food insecurity among young Cambodian women in Massachusetts.Methods: A cross‐sectional survey was conducted with young women of Cambodian heritage ages 15‐30 years of age (n=127). The dependent variable, household food security score, was measured using the USDA 6‐item module. The independent variable, depression score, was ascertained using the Harvard Trauma Scale. Multivariable linear regression was used to determine the association between depression status and household food security score, controlling for age and pregnancy status.Results: Of 127 women, 25.6% lived in low/very low food secure households. The mean depression score was 23.8. Lower depression scores were observed among pregnant women compared to non‐pregnant women (24.3 vs. 30.9, p<0.05). Women with higher depression scores were more likely to be food insecure than women with lower depression scores (β ‐0.475; 95% CI: ‐0.949, ‐0.002), after controlling for age and pregnancy status.Conclusion: Household food security scores were positively associated with depression among Cambodian young women. This finding suggests the role of depression as a potential indicator of household food security status. Future research on these associations among refugee women living in the United States are warranted.Grant Funding Source: USDA
Identification and clinical prognosis of salinomycin binding targets in neuroblastoma (LB613)
Authors: Zhou Shuang , Wang Fengfei , Xiang Shihua , Wong Eric , Muhonen Wallace , Fonkem Ekokobe , Hsieh Tze‐chen , Li David , Zhang Ruiwen , Shabb John , Wu Joseph , Wu Min , Wu Erxi 
Published: June 2021

Salinomycin, a widely used anti‐coccidial agent, was recently identified from a library of 16,000 natural and commercial chemical compounds based on its highly selective inhibitory effect on breast cancer stem cells (CSCs), with more than 100‐fold greater potency than paclitaxel. Salinomycin also exhibits cytotoxic effects on other types of cancer cells and CSCs and overcomes drug resistance. However, the exact mechanism of salinomycin, especially its direct binding target(s), and its effects on Neuroblastoma (NB) are yet not known.NB is a common solid tumor and a leading cause of mortality in children. Currently, 35% of patients with NB remain incurable. In addition, the majority survivors of NB suffer from long‐term side effects of current therapies and are at risk for disease relapse or getting a second, different cancer. More effective therapies are pressingly needed. Since the existence of CSCs in human NB cell lines and NB tumors has been well documented, and has been closely associated with chemoresistance or tumor relapse, therapeutic targeting of NB CSCs may be a critical novel approach for NB therapy.Aiming to improve NB therapy, we examined the efficacy and mechanism of salinomycin in human NB cells. Our study showed that salinomycin markedly inhibits NB cell proliferation and tumorsphere formation. Treatment of salinomycin induced G2 cell cycle arrest with an up‐regulation of Cyclin A and a down‐regulation of p21 protein levels. We further identified TIF1β and NCL as novel direct binding targets of salinomycin by using comprehensive methods, including chemical proteomics and functional genomics. Furthermore, by analyzing tumor samples from a cohort of 88 NB patients, we found that the elevated level of either TIF1β or NCL is associated with poor prognosis for NB patients. Thus, our results indicate salinomycin is a potential new therapeutic agent and TIF1β and NCL as new therapeutic targets/biomarkers for NB therapy.Grant Funding Source: Supported by ND EPSCoR, NDSU, NIH P20 RR020151, NIGMS (P20 GM103505 and P30 GM103332‐01)
ACTH administration stimulates both cortisol and aldosterone secretion in fasting northern elephant seals (LB776)
Authors: Champagne Cory , Tift Mike , Houser Dorian , Crocker Daniel 
Published: June 2021

During molting, northern elephant seals exhibit substantial alterations in circulating cortisol concentrations, probably to facilitate pelage synthesis while fasting. How these changes influence the hypothalamic‐pituitary‐adrenal (HPA) axis and the stress response, however, is not known. To investigate the modulation of the HPA axis during molting, we administered ACTH (25 units, intramuscularly) to sixteen juvenile elephant seals at the beginning (n=6), middle (n=5), and end (n=5) of their spring molting period in a cross‐sectional study design. Blood samples were collected immediately before, and then periodically for 2.5 hrs after, ACTH administration. We quantified circulating concentrations of ACTH, and the responses of cortisol and aldosterone. Both cortisol and aldosterone concentrations markedly increased following stimulation by ACTH and remained elevated for the duration of sampling (linear mixed model, F = 153, 25.3, respectively; p < 0.01). There were no detectable differences in the magnitude of hormone responses to ACTH administration among the study groups (p > 0.1). Nonesterified fatty acids increased following ACTH administration, suggesting upregulation of lipolysis; however, blood urea nitrogen concentrations increased only marginally, indicating that protein catabolism was less affected by ACTH administration. ACTH is not a strong aldosterone secretagogue in terrestrial mammals but several studies have measured increased aldosterone after ACTH stimulation in marine mammals, indicating its potential importance in osmoregulation during stress in these species. Cortisol and aldosterone increased in parallel following ACTH stimulation but the slope of this relationship varied among the study groups. This suggests that the release of these hormones is closely coupled in this species and possibly other marine mammals.Grant Funding Source: Supported by the Office of Naval Research
Biochemistry and Molecular Biology Education in a Transforming Academy and a Molecular World
Authors: Brenner Charles 
Published: June 2021

Every year, we welcome 3 million new undergraduates to US colleges and universities, approximately one in six of whom begin their collegiate careers with premedical intentions. Thus, approximately 500,000 students annually enroll in inorganic chemistry. However, only about 165,000 go on to take organic chemistry, and in 2008, only 42,200 applied for 18,000 slots in US medical schools, thereby producing 17,300 new US MDs in 2012. This premedical funnel has a huge effect on the organization of undergraduate colleges and the education of nearly half a million undergraduates per year who are not going to become doctors. The redesign of the 2015 MCAT was therefore an opportunity not only to think about what courses premedical students ought to take but to consider how to prepare larger populations of undergraduates with foundational molecular knowledge. Premedical students now almost universally have access to a biochemistry course either at their home campus or online. Accordingly, while medical schools are admitting students for fall 2016 enrollment who have largely taken and been examined in biochemistry and molecular biology, the basic science coursework in the first two years of medical school has simultaneously been condensed in a manner that makes it difficult to teach metabolism and genomics at the advanced levels that might have been enabled by a better prepared entering class. In this talk, I will address approaches to offer molecular education to undergraduate and health professional students that have the potential to improve our students’ appreciation of data analysis, drug and biomolecule reactivity, enzymes, metabolism and genomics.Support or Funding InformationEducational activities supported by National Science Foundation award CHE1404147 and the Roy J. Carver Trust.
Mechanosensory function of Cav1.2 calcium channel in renal primary cilia
Authors: Kathem Sarmed H , Nauli Surya M 
Published: June 2021

Primary cilia are mechanosensory organelles and play an important role in calcium signaling. However, the precise mechanistic event of cilia‐calcium axis is still debated. Here, we used kidney cells transfected with G‐CaMP3 (a green fluorescent genetically encoded calcium indicator) to assess this important event in cell function. Using modified single‐cell imaging technique, we were able to capture a single‐living cell from the side for a better viewing of the cell body and its cilium simultaneously. Recently, we also used a high‐speed imaging recording (a minimum of 200 frames per sec) to capture both cilia bending and calcium signaling with Nomarski interference contrast (NIC) with fluorescence microscopy, respective. Within 1.5 μsec after fluid‐shear induce cilia bending, we reported calcium current was detected at the cell membrane very near to cilium base. This calcium current was directionally organized with the fluid flow and cilia bending direction. When ATP was added to the cells without flow, directionally non‐organized calcium current was seen throughout the cell membrane and cytosol. Immunofluorescence staining showed that Cav1.2 calcium channel expressed at the cell membrane close to cilia base. Verapamil completely abolished flow‐induced calcium signals suggesting that Cav1.2 channel might be responsible for calcium signaling. Calcium‐depleted extracellular fluid with EGTA produced a similar result as to verapamil indicating that extracellular calcium was required to initiate this event. We conclude that the very early event in cilium‐induced calcium signaling initiated in the cell membrane close to cilium base through Cav1.2 calcium channel.Support or Funding InformationDepartment of Defense (PR130153 )
Maternal Vitamin A Supplementation Expands Adipose Progenitor Population through Promoting Vascular Development
Authors: Wang Bo , Fu Xing , Zhang Shuming , Liang Xingwei , Zhu Meijun , Du Min 
Published: June 2021

Retinoic acid regulates various types of progenitor differentiation and tissue development including adipose development. However, the impact of maternal vitamin A supplementation on fetal adipose development and its long‐term consequence in adulthood remain unclear. To address, we supplemented maternal mice with 0, 15 or 30 IU/mL retinyl palmitate in water during gestation and lactation. Adipose tissue and serum of weanling offspring, and E7.5, 12.5 and 18.5 fetuses were collected. We found that retinoic acid is the main retinoid in fetuses that was elevated by maternal vitamin A supplementation. The populations of platelet‐derived growth factor receptor alpha positive (PDGFRα+) adipocyte progenitors in both fetuses and weaned offspring were increased due to vitamin A supplementation, which were correlated with heightened vascular density in adipose tissue at weaning. We further found that retinoic acid upregulated vascular endothelial growth factor α (VEGFα) which might explain the enhanced vasculogenesis in adipose tissue of fetuses supplemented with vitamin A. Using florescence tracing, we found that, under cold or retinoic acid stimulation, PDGFRα+ progenitor cells were located in neovascular system. In summary, our results reveal that maternal vitamin A supplementation increases offspring adipose progenitor populations by promoting vascular system development during the fetal stage, which is mediated by retinoic acid receptor signaling.Support or Funding InformationSupported by NIH R01HD067449 and USDA‐NIFA 2015‐67015‐23219
Unjamming and cell shape in the asthmatic airway epithelium
Authors: Fredberg Jeffrey , Mitchel Jennifer , Kim Jae Hun , Park Jin‐Ah 
Published: June 2021

From coffee beans flowing in a chute to cells remodeling in a living tissue, a wide variety of close‐packed collective systems –both inert and living– have the potential to jam. The collective can sometimes flow like a fluid or jam and rigidify like a solid. The unjammed‐to‐jammed transition remains poorly understood, however, and structural properties characterizing these phases remain unknown. Using primary human bronchial epithelial cells, we show that the jamming transition in asthma is linked to cell shape, thus establishing in that system a structural criterion for cell jamming. Surprisingly, critical scaling collapse predicts a counterintuitive relationship between jamming, cell shape, and cell‐cell adhesive stresses that is borne out by direct experimental observations. Cell shape thus provides a rigorous structural signature for classification and investigation of bronchial epithelial layer jamming in asthma, and potentially in any process in disease or development in which epithelial dynamics play a prominent role.Support or Funding InformationThis research was supported by Francis Family Foundation, the Alfred P. Sloan Foundation, American Heart Association (13SDG14320004), National Research Foundation of Korea (NRF‐2013S1A2A2035518), National Science Foundation (BMMB‐1334611, DMR‐1352184), and the National Institutes of Health (K25 HL091124, P30DK065988, P30ES000002, HL007118, R01HL102373, R01HL107561, P01HL120839)
Tannase‐treated grape pomace attenuates IL‐1β‐induced inflammation in Caco‐2 cells
Authors: Martin Isabela , Macedo Gabriela , Blumberg Jeffrey , Chen C‐Y. Oliver 
Published: June 2021

Grape pomace (GP) derived from wine production is rich in glycosidic phenolics which may possess a lower bioefficacy than their respective aglycones. Thus, we examined whether GP treated with Paecilomyces variotii tannin acyl hydrolase (tannase) to hydrolyze the ester and the depside bonds between conjugates and phenolics enhances anti‐inflammatory actions in vitro. GP was treated with tannase at 40 °C and pH 5.5 for 5 h. Phenolics in GP and tannase treated GP (GPT) were extracted with 70% methanol in acetate buffer, dried under N2 air, and reconstituted in PBS for tests. GPT contained 45% more quercetin and 71% less quercetin‐3‐O‐rutinoside than GP. Further, GPT had 39% more total phenols than GP and displayed 57, 215, and 12% greater total antioxidant capacity per DPPH, ORAC, and FRAP analyses, respectively. However, GP and GPT at 100 and 200 μg/mL (dry extract wt/v) displayed comparable efficacy in the reduction of ROS production in Caco‐2 cells treated with or without 50 μM AAPH. After 24‐h pretreatment, GPT at 200 μg/mL decreased 20 ng/mL IL‐1β‐induced PGE2 production by 110% and IL‐8 by 83% and down‐regulated NF‐κB activation by 68% relative to controls. GPT was more potent than GP in the amelioration of IL‐1β‐induced inflammation. Therefore, tannase treatment appears to enhance the antioxidant and anti‐inflammatory activities of grape pomace phenolics and suggests the potential application of this material in food and beverage products.Support or Funding InformationSupported by USDA
Hepcidin attenuates zinc efflux in Caco‐2 cells
Authors: Hennigar Stephen R. , McClung James P. 
Published: June 2021

Hepcidin, a peptide hormone produced by theliver in response to iron loading and inflammation, regulates body iron storesby inhibiting iron transporter function. Zinc also regulates hepcidinexpression; however, the effects of hepcidin on zinc transport and zinctransporter function have not been documented. The objective of this study wasto determine whether hepcidin affects intestinal zinc transport. Differentiatedhuman intestinal Caco‐2 cells were treated ± 1μM hepcidin for 3–24 h. Zinctransport was assessed in cells seeded on Transwell inserts. Media was collected from the apical and basolateral chambers and 67Znconcentrations were determined by ICP‐MS. Expression of metallothionein (MT1A), ZIP4 (SLC39A4), and ZnT1 (SLC30A1)was determined using RT‐qPCR and Western blot. Hepcidin attenuated zinctransport, as cells treated with hepcidin exported 20 and 26% less 67Zninto the basolateral chamber at 3 h and 24 h (P<0.05), respectively, and retained more cellular 67Zn(P<0.05) as compared to control cells. Intracellular zinc in control cells was localized to vesicles dispersed throughout the cytoplasm; however, the number of vesicles and intensity of FluoZin‐3 fluorescence decreased (P<0.05)in cells treated with hepcidin for 24 h. The mRNA abundance of metallothionein increased 1.5‐fold (P<0.05). ZIP4 protein remained unchanged (P>0.05), whereas hepcidin reduced ZnT1 protein by 75% (P<0.05) as compared to control cells. These data suggest that hepcidin is a regulatory hormone that manages zinc transport in intestinal cells by downregulating ZnT1 to reduce zinc export and increasing metallothionein to buffer the rise in intracellular zinc. Understanding the mechanism and role of hepcidin in zinc homeostasis may provide insight into the hypozincemiaassociated with inflammation and infection.Support or Funding InformationSupport provided by USAMRMC.
Skin Carotenoids as Biomarker for Vegetable and Fruit Intake: Validation of the Reflection‐Spectroscopy Based “Veggie Meter”
Authors: Ermakov Igor V , Whigham Leah D , Redelfs Alisha H , Jahns Lisa , Stookey Jodi , Bernstein Paul S , Gellermann Werner 
Published: June 2021

Skin is a relatively stable storage medium for carotenoids; non‐invasive optical measurements of carotenoids in this tissue via Resonance Raman spectroscopy (RRS) serve as a non‐invasive biomarker for fruit and vegetable (F/V) intake. The RRS method has been validated with HPLC‐based measurements of carotenoid concentrations in serum and excised skin tissue samples and has been shown in feeding studies to respond to changes in F/V intake.An attractive alternative to RRS is based on reflection spectroscopy (RS), which requires less complex instrumentation and has an advantage in also measuring skin melanin and hemoglobin absorptions. Using a suitable algorithm, the latter can be taken into account in the RS‐based derivation of skin carotenoid levels. Furthermore, these levels can be quantified in optical density units, which directly correlate with the carotenoid concentrations encountered in the measured tissue volume. The objective of this study was to validate the RS method via comparison with the RRS method.We compared RS and RRS measurements in 5 diverse human subject cohorts of different age groups and ethnicities in community and clinical settings. These measurements included inter‐subject comparisons of carotenoid levels in 146 adults and children, tracking of carotenoid changes in response to dietary interventions in 29 adults, and an ongoing exploration of a potential correlation between skin and retinal carotenoid levels in adult patients with vision disorders. Depending on the particular study, correlation coefficients varied between R ~ 0.80 and 0.96, showing that the RS method provides a valid substitute for RRS in skin carotenoid measurements of all age groups. A compact, portable instrument configuration will be presented, which permits rapid non‐invasive measurement and tracking of F/V derived carotenoid levels. In its capacity as “Veggie Meter” it should be attractive for screening of large populations and the assessment of the effectiveness of F/V intake interventions.Support or Funding InformationSupport for this research was provided in part by the Paso del Norte Health Foundation and the USDA‐ARS.
Menstrual cycle hormones, food intake, and cravings
Authors: Krishnan Sridevi , Tryon Rebecca , Welch Lucas C , Horn William F , Keim Nancy L 
Published: June 2021

ObjectiveFood craving and intake are affected by steroid hormones during the menstrual cycle, especially in the luteal phase, when craving for certain foods has been reported to increase. However, satiety hormones such as leptin have also been shown to affect taste sensitivity, and therefore food choices. While in vitro studies have reported that estradiol can stimulate leptin release, some human studies have shown that leptin is inversely associated with estradiol. On the contrary, progesterone has been positively associated with leptin, and therefore may play a significant role in food intake regulation. Our objective was to characterize the associations between sex steroid hormones, leptin and food intake behavior.MethodsSeventeen healthy, women with regular menstrual cycles, 23.2 ± 4.8 y, and BMI 22.4 ±1.9 kg/m2 were studied during their late follicular and luteal phases. Estradiol, progesterone, DHEAS, SHBG, leptin and insulin were measured in fasting samples. In addition, volunteers were asked to answer validated “Food Craving inventory” (FCI) and “Carbohydrate Rich Food intake” (CRFI) questionnaires at the start of the study. The FCI captured information regarding the types of foods volunteers were craving – rich in fat, carbohydrate or sweet taste, followed by beverages and fast foods over a three month period prior to starting the study. The CFRI captured information with regard to consumption of these same foods. We used Pearson's correlation coefficients to identify positive or inverse associations between hormones and subjective food intake and craving scores.ResultsEstradiol, progesterone and leptin were not associated with each other. However, women with higher estradiol during the luteal phase reported consuming more carbohydrate‐rich foods in the past three months (r = 0.5174; p = 0.034), while also reporting increased craving for sweet foods (r = 0.4922; p = 0.045). Higher progesterone in the luteal phase was weakly associated with higher craving for sweetened beverages (r = 0.4723, p = 0.056). As expected, leptin was inversely associated with reported consumption of sweet tasting foods (r = −0.5071; p = 0.038), while also being inversely associated with craving fat rich foods (r = −0.4851; p = 0.048).ConclusionsEstradiol is associated with increased carbohydrate rich food intake, while progesterone with craving sweetened beverage intake. Leptin, while being associated with reduced craving for fat‐rich food, and reduced sweet food intake, however, appears to not be associated with estradiol and progesterone, thereby suggesting that the female sex steroid hormones may be independent influences on food intake behaviors. Understanding the changes of estradiol, progesterone and leptin in women could clinically aid in advocating food choices during the pre‐menstrual phase.Support or Funding InformationUSDA projects #2032‐51530‐022 and #5306‐51530‐019