bluuebunny/crossref_metadata_2025_split huggingface dataset:
CDKN1C (P57KIP2) NEGATIVELY REGULATES E2F1‐DRIVEN TRANSCRIPTION VIA A DIRECT INTERACTION
Authors: Cress Doug , Wright Gabriela M , Ma Yihong 
Published: June 2021

ObjectiveTo determine the mechanism by which CDKN1C binds and transcriptionally represses E2F1‐driven transcription. We previously found that E2F1 can induce transcription of the cyclin‐dependent kinase inhibitor CDKN1C (p57, KIP2). Furthermore, we showed that the CDKN1C protein could in turn repress E2F1‐driven transcription. In the current work, we explore the mechanism by which CDKN1C represses E2F1‐driven transcription. We demonstrate that E2F1 and CDKN1C can form stable complexes both in vivo and in vitro. Molecular studies demonstrate that the E2F1/CDKN1C interaction is mediated by two E2F domains. A central E2F1 region containing the "marked box" domain interacts directly with CDKN1C, whereas a C‐terminal E2F1 domain interacts with CDKN1C via interaction with Rb. Chromatin immunoprecipation assays demonstrate that CDKN1C associates with known E2F1‐regulated promoters in vivo. Overexpression of CDKN1C results in an inhibition of RNA polymerase II C‐terminal domain phosphorylation suggesting a mechanism by which promoter‐associated CDKN1C may inhibit transcription. These results highlight a novel mechanism by which E2F1 transcriptional activity is restrained.
Dinitrobenzene Isomers Structure‐ Dependent Stimulation of Neuronal Nitric Oxide Synthase Catalyzed Electron Flux in the Absence of Calmodulin
Authors: Joshi Chintamani N , Miller R Timothy 
Published: June 2021

Neuronal nitric oxide synthase (nNOS) requires calmodulin (CaM) ‐ binding for efficient electron transfer converting L‐arginine to L‐citrulline and nitric oxide. The present studies focus on electron transfer to and through CaM‐free nNOS. Turnover rates (min−1) of nNOS‐mediated NADPH oxidation (NADPHox) and acetylated cytochrome‐c reduction (AcCyt‐cred) were used to estimate total electron flux and O2. production. Dinitrobenzenes (DNBs) stimulated the rate of NADPHox by CaM‐free nNOS in an isomer‐ and concentration‐ dependent manner. Rates for NADPHox by CaM‐free nNOS at the limit of solubility of the DNBs (160 ?M) were 189 ± 5 min−1, 94 ± 3 min−1 and 38 ± 2 min−1 for 1,4‐, 1,2‐ and 1,3‐DNB, respectively, compared to the solvent control (7 ± 1 min−1). Highly stimulated rates of AcCyt‐cred were observed in presence of all the three DNBs. The nNOS reductase construct without the CaM binding domain (aa 745‐1729) behaved in a manner similar to holo‐nNOS in the presence of DNBs, in the NADPHox and AcCyt‐cred assays. These results indicate that CaM‐free nNOS generates ROS at environmentally‐relevant levels of DNBs (ASTDR). Considering the absolute amount of constitutive NOS expressed in organisms; chronic exposure to low levels of DNBs may lead to continuous oxidative stress.Supported by ES011982 and RR008124 to RTM and UTEP respectively.
Intein‐mediated protein splicing of human Eg5 kinesin
Authors: Korona Dagmara , Korona Boguslawa , Wojcik Edward 
Published: June 2021

HsEg5 is a homotetrameric BimC/kinesin‐5 family member that plays a vital role in the mitotic spindle. This crucial role has identified HsEg5 as a promising target for cancer chemotheraphy.The aim of a project is to clarify the role of the sequence regions in kinetic mechanism of HsEg5 and to determine the various conformational states in which this protein occurs while performing its biological function. Intein‐mediated protein ligation (IPL) was used as a tool for segmental labeling of proteins of interest. Based on various criteria including the presence of cysteine required for peptide bound formation (native or artificially introduced), the breaking points for IPL were designed, so that the separately expressed fragments of protein could be joined together. The rebuilt protein has been obtained in a very high yield compared to values reported in the literature. The resulting IPL product reconstructed from fragments exhibits catalytic activity of native enzyme, however, due to the amino acid exchange (substitution for cysteine), the activity differs from that of wild type enzyme. The segmentally labeled protein is then subjected to analysis by Fourier‐Transform Infrared Spectroscopy (FTIR).
Murine Genotype Alters Palatal Tissue Response to Nicotine
Authors: Serrano Maria Juliana , Yu Wenli , Svoboda Kathy 
Published: June 2021

Our previous work demonstrated that nicotine inhibited palatal fusion in vitro in a dose dependent manner. We asked if the mouse genotype would be a factor in nicotine teratology.METHODSPalatal shelves from mouse strains (CD1, ND4, C56 and 129S) E13.5 were cultured in serum free media (control), and nicotine (0.6 mM and 6 mM). Palates were harvested at 24 hours, 48 hrs and 72 hrs and fixed in 4%PFA for paraffin embedding and H&E sections.RESULTSCD1 and ND4 strains were consistent in the palatal fusion pattern within litters. The shelves in all the embryos are not fused at a specific age (E 13.5).C56 and 129Shad variable palate fusion. Some of the embryos had fused palates at embryonic day 13.5, while other embryos had undeveloped palatal shelves. The tissue cultured palates from CD1 and ND4 mice treated with 0.6mM nicotine fused, while palate treated with 6 mM nicotine retained epithelial seam. Palates from C56 mice had an epithelial seam using low and high doses of nicotine. 129S palate shelves 6mM nicotine have two layers of MEE in the midline (non fusion).CONCLUSIONSThe genetic background of mice influenced nicotine responses in palatal tissues.Grant Funding Sources
Effects of Nonstructural Proteins (NS) of Respiratory Syncytial Virus (RSV) on Host Interferon Response
Authors: Swedan Samer Fuad , Barik Sailen 
Published: June 2021

RSV is a Pneumovirus of the Paramyxoviridae family. To replicate efficiently, RSV interacts with the host cell to inhibit cellular innate immunity. RSV has two nonstructural proteins, NS1 and NS2, which have no homology with any other gene in biology. Both are dispensable for RSV replication in cell culture, but needed for robust virus infection in animals. Both NS proteins have interferon (IFN) antagonist activity. The project's goal is to define the functions of the NS1 and NS2 proteins of RSV in viral replication and pathogenesis. More specifically, the amino acid residues of the NS proteins essential for IFN antagonism will be identified and host proteins that interact with the NS proteins will be determined. The central hypothesis is that NS1 and NS2 antagonize IFN production by interacting with components of IFN induction and response cascades. To test this hypothesis, techniques such as site‐directed mutagenesis of NS genes, luciferase reporter assays, and IFN functional assays will be employed. This will provide an integrated view of the structural determinants of NS proteins on IFN antagonism and the effects of altering these determinants on RSV biology. Initial results indicate that NS1 and NS2 prevent the induction of IFN‐β and implicate the ten c‐terminal residues of NS2 in the degradation of STAT2.Research sponsored by Alabama EPSCoR and NIH Grant AI059267
Charactrization of anti‐aging effect in Halophyte extracts, Suaeda asparagoides
Authors: Hong Heeok , Ahn Jeung‐Youb 
Published: June 2021

The aim of this research was to develop anti‐skin aging product using anti‐oxidant effective materials in halophyte plant, Suaeda asparagoides (SA Ex).We tried out several experimental works in in vitro and in vivo (animal study and human clinical study). To find the anti‐oxidant effects in this study we used DPPH, photo‐hemolysis, luminol‐dependent chemiluminescence assay and radical scavenging assay. Anti‐oxidant effect shown 87.8% free radical scavenging activity in ethylacetate fraction of SA Ex. Biochemical enzyme inhibition assay was done by collagenase, hyaluronidase, and elastase. Among the Collagenase, hyaluronidase and elastase inhibition activity, collagenase inhibition activity was shown strong effect definitely. MTT assay was employed for cell cytotoxicity effect. We also investigated formula stability and stabilized formula was applied to in vivo test which was the skin sensitization test on hairless mouse (HR‐1).In conclusion, first, we have got outcome material registration on International Cosmetic Ingredient Dictionary (ICID). Second, this research report is first of all about isolation of components in Suaeda asparagides. Third, we made guarantee that is data on material safety and stability by in vitro and in vivo.These anti‐oxidative fraction and materials could be re‐evaluated the biological effects on research area. The data show us potential anti‐aging biological ability and the materials were expected as a functional ingredient of industrial products in the future.
Dissecting the function of the HigB addiction module toxin
Authors: Hurley Jennifer Marie , Woychik Nancy Ann 
Published: June 2021

Toxin‐antitoxin (TA) modules were initially discovered on bacterial plasmids that facilitate plasmid maintenance via post segregational killing. More recently, chromosomally encoded TA operons found in free‐living prokaryotes have been demonstrated to aid in the cellular response to stress. TA modules encode a stable toxin that inhibits cellular growth, and a labile cognate antitoxin, that inhibits the action of the toxin through the formation of a toxin/antitoxin complex. In addition, these modules have been implicated in pathogenesis and antibiotic tolerance. TA systems also represent a new class of targets for antibiotic treatment of pathogens resistant to existing therapeutics.The goal of this project is to identify the precise function and intracellular target of the HigB toxin, which is a member of the RelE family of toxins. When expressed, RelE or HigB inhibit protein translation through mRNA cleavage that is mediated by the association of the toxins with the Escherichia coli 50S ribosomal subunit. However, the specificity of mRNA cleavage and the nature of the ribosome‐dependent toxin activity appear to be distinct between the two toxins. We are currently investigating additional mechanistic details of HigB toxicity. Research supported by the Virus‐Host Interactions in Eukaryotic Cells T32 Training Grant from the NIH‐NIAID awarded to Dr. Sidney Pestka.
Studies on the Effect of Urea Application on Nitrate Toxicity in Forages
Authors: Chaudhary Dharam Paul , Tiwana Udham Singh , Bhardwaj Behari Lal , Puri Krishan Pal , Muker Harvinder Singh 
Published: June 2021

ObjectiveUnder certain adverse conditions nitrate can accumulate to potentially toxic concentrations in forages like oats (Avena sativa), barley (Hordeum vulgare), maize (Zea mays), sarson (Brassica juncea) and berseem (Trifolium alexandrinum). The conditions under which plants accumulate excessive amounts of nitrates are not fully understood. We hypothesized that excessive use of nitrogenous fertilizers may be responsible for accumulation of nitrate in plants and decided to determine the effect of excess urea application on nitrate accumulation in berseem, berseem+ rye grass (Lolium perenne) (B+RG), breseem + sarson (B+S) and berseem + oats (B+O).MethodsThe crop was sown on the experimental area in the plot sizes of 5X4 m2 in three replications. The crop was sprayed with 5 different concentrations of urea ranging from 1 to 5% and subsequently analyzed for nitrate content.ResultsThe results indicate that the crude protein and nitrate content of berseem, B+RG, B+S and B+O increases with increasing concentrations of urea. The maximum nitrate concentration in B+S was observed at 4% of urea concentration while in B+RG and B+O at 5% of urea.ConclusionThe excessive nitrogen application appears to be responsible for the nitrate accumulation in the forage crops which may lead to large scale livestock losses.
Energy‐induced changes in skeletal muscle protein synthesis and molecular signaling in physically active adults
Authors: Pasiakos Stefan M , Vislocky Lisa M , Carbone John W , Freake Hedley C , Anderson Jeffrey M , Ferrando Arny A , Wolfe Robert R , Rodriguez Nancy R 
Published: June 2021

To date, no studies have characterized how energy deprivation influences skeletal muscle protein utilization.PURPOSEThis investigation characterized the effect of an acute energy deficit on mixed muscle protein synthesis (MPS) and associated intracellular signaling proteins (ISPs) in physically active adults.METHODSEleven men and seven women participated in a 20‐d, two‐phase diet intervention study: weight maintenance, WM; and energy deficient, ~80% of estimated energy requirements, ED. Dietary protein (1.5 g·kg−1·d−1) and fat (~30% of total calories) were constant for WM and ED. MPS and ISPs were measured on day 10 of both interventions using a primed, constant infusion of [2H5]‐phenylalanine and Western blotting techniques, respectively. Resting energy expenditure (REE) was estimated using indirect calorimetry and nitrogen balance (NBAL) was determined.RESULTSSubjects lost ~1 kg body weight during ED (P < 0.0001). REE or NBAL were not affected. MPS was reduced ~33% (P < 0.05) for ED (0.056 ± 0.01 %/h) compared to WM (0.083 ± 0.007 %/h). Akt and 4E‐BP1 phosphorylation were lower (P < 0.05) during ED compared to WM.CONCLUSIONThese findings show that MPS and associated synthetic ISPs are downregulated in response to an acute, moderate energy deficit in physically active adults and provide a basis for studies assessing the impact of prolonged energy restriction on skeletal muscle protein metabolism.Grant Funding SourceNational Cattlemen's Beef Association
Dietary Docosahexaenoic Acid Fully Restores Fertility and Spermiogenesis in the Male Delta‐6‐Desaturase Knockout Mouse
Authors: Roqueta‐Rivera Manuel , Stroud Chad K. , Segre Mariangela , Haschek Wanda M. , Hess Rex A. , Agbaga Martin‐Paul , Nakamura Manabu T. 
Published: June 2021

Delta‐6 desaturase (D6D) is the rate limiting enzyme for the synthesis of highly unsaturated fatty acids (HUFAs): 20:4n6 arachidonic acid (AA), 22:6n3 docosahexaenoic acid (DHA), and 22:5n6 docosapentaenoic acid (DPA), all of which are abundant in murine spermatozoa. The D6D knockout (‐/‐) mouse, unable to synthesize HUFAs, is infertile by 6 weeks of age due to disrupted spermiogenesis and lack of spermatid elongation. In order to determine which HUFA is essential for spermiogenesis, AIN93G diet supplemented with either 0.2 % (w/w) AA or DHA was fed to wild type (+/+) and D6D ‐/‐ mice at weaning until 16 weeks of age (n=3‐5). Frequency of successful matings of DHA supplemented ‐/‐ (61%) was not statistically different vs +/+ on DHA. The success rate of AA supplemented ‐/‐ (38%) was lower though not significant vs +/+ on AA, but higher than non‐supplemented ‐/‐ (8%, p<0.05). DHA supplemented ‐/‐ had normal sperm count (116% of +/+) and motility, while AA supplemented ‐/‐ had normal sperm motility but significantly lower sperm count (39% of +/+, p < 0.05). Histology of DHA supplemented ‐/‐ showed restored spermiogenesis with mature sperm in epididymis, while AA supplemented ‐/‐ had partially restored spermatogenesis with mature and immature sperm in epididymis. In conclusion, dietary DHA fully restores male reproductive function in ‐/‐. Partial restoration by AA may be due to sparing of DPA from retroconversion to AA.Grant Funding SourceInternal
Galactose promotes fat mobilization in obese lactating and non‐lactating women
Authors: Mohammad Mahmoud A , Sunehag Agneta L , Rodriguez Luisa M , Haymond Morey W 
Published: June 2021

BackgroundGalactose consumption results in a lower rise in plasma glucose and insulin concentrations when compared to glucose. The lower insulin might promote lipolysis during meal absorption.HypothesisAn isocaloric galactose drink, when compared to glucose, will sustain fat mobilization during meal consumption while not adversely affecting milk production.Subjects5 healthy, obese (BMI 34.4±2.4), exclusively breastfeeding and 5 non‐lactating (BMI 36.3±1.3) women were studied in a crossover, randomized, single blinded design. During 2d on both occasions, subjects received drinks providing 70% of the daily resting energy requirement (RMR) of which glucose or galactose provided 55% of RMR. Primary outcome variables were: rates of appearance (Ra) of glycerol, glucose and palmitate, milk production, substrate and hormone concentrations, energy expenditure and substrate oxidation.ResultsIn all subjects, during galactose, as compared to glucose feeding, plasma glucose, insulin and C‐peptide were lower (p<0.05) while FFA and glycerol were higher (p<0.01). Glucose Ra was 72% lower and glycerol and palmitate Ra were 39 and 53 % higher (p<0.01) during galactose consumption. Glycerol Ra ( μmol·kg−1·min−1) was higher during galactose than glucose feeding in both control (2.0±0.3 vs. 1.4±0.2, p<0.05) and lactating women (2.2±0.4 vs. 1.7±0.3, p<0.01). Milk production, energy expenditure and substrate oxidation were similar between the two sugars.ConclusionsConsumption of galactose sustains fat mobilization during meal absorption. Thus; galactose might facilitate body fat loss in obese women. RO1DK 55478, HD 37857, MO1‐RR‐00188‐34.
Diabetes in HIV Positive Adults in Botswana: Nutritional and Demographic Characteristics
Authors: Hatsu Irene , Baum Marianna K , Huffman Fatma , Bussmann Hermann , Dusara Priti , Makhema Joe , Burns Patricia J , Sales Sabrina , Campa Adriana , Marlink Richard 
Published: June 2021

ObjectiveBotswana is an African country with one of the highest rates of HIV (24.1%) in the general population, in the world. The objective of this study was to determine the prevalence and characteristics of diabetics among HIV positive adults in the early stages of the disease who are not on antiretroviral medication.MethodsAfter obtaining informed consent, 610 HIV‐positive asymptomatic patients were recruited for the study. The diagnosis of diabetes was obtained from medical records. Structured clinical, demographic and physical activity questionnaires were administered. Two 24‐hour recalls were averaged for each participant.ResultsOut of the 610 recruited, 28 (4.6%) were diabetics compared to 7.2% for the general population in Botswana. In those aged 40 years and older, however, the rate of diabetes in this cohort was 10.8%. Compared with non‐diabetics (n=582), diabetics were older (39.3 y vs. 33.3 y, p<0.001), had higher BMI (27 vs. 24 kg/m2, p=0.01) and were less physically active (p=0.002). There were no statistical differences in intakes of carbohydrates, fat and total calories between the two groups.ConclusionAsymptomatic HIV infection doesn't seem to affect the prevalence of diabetes in this population. Risk factors for the development of diabetes are similar in this HIV+ cohort to those for the general population.Supported by NIDA
Diet, Health factors Cuban Americans w/wo T2DM
Authors: Huffman Fatma G. , Vaccaro Joan A , Zarini Gustavo , Nath Subrata 
Published: June 2021

ObjectiveTo assess dietary, anthropometric and clinical predictors of cardiovascular disease (CVD) by gender and diabetes status in Cuban Americans.MethodsCross‐sectional, case control, study of aged‐matched, Cuban American adults with and without diabetes, randomly selected by a mailing list from three counties in South Florida resulted in an informed consent study for whom N= 367 190 diabetics (72 males, 118 females) and 177 non‐diabetics (59 males and 118 females) furnished complete data.ResultsANOVA confirmed group differences by age, gender and diabetic status in diet and clinical predictors of CVD. Regression confirmed % carbohydrate intake was associated with fasting glycated hemoglobin A1c (HbA1c) for non‐diabetic females, exclusively.ConclusionsDifferences in diet by diabetes status and gender were associated with health outcomes. Diabetes management of glucose control was associated with age.
Food and Fun Curriculum for Latina Mothers and Children
Authors: Racine Elizabeth Fussell , Coffman Maren , Chrimes Dale 
Published: June 2021

Latina women and children suffer from increased rates of obesity, poor nutrition, and decreased physical activity. In order to address these concerns, the Food and Fun curriculum developed by the Harvard School of Public Health was adapted to educate Latina mothers and children about nutrition and physical activity.A pre and post test study design was utilized. The program consisted of an eight week educational and exercise intervention. Before and after the intervention, an assessment of BMI, waist circumference, fruit and vegetable intake, and physical activity level were taken from mothers and children.Pre‐intervention data were collected on 15 women; 7 women completed the study. According to paired samples t‐test, there was a significant increase in fruit and vegetable consumption in mothers (t(6) = ‐3.0, p < .02). Though not statistically significant, amount of time engaged in physical activity increased from Time 1 (M = 592.9) to Time 2 (M = 910.0). Analysis of children's data is ongoing.The study provided preliminary data on the effectiveness of the adapted Food and Fun curriculum for low income Latina mothers and children in Concord, NC. Given the growing Latino population and increasing obesity rate, it is important to test culturally appropriate nutrition and physical activity interventions that have the potential to improve the health of countless vulnerable individuals.
Zinc stimulates extracellular matrix mineralization in osteoblastic MC3T3‐E1 cells
Authors: Alcantara Ethel Hipolito , Lomeda Ria‐Ann R , Cho Young‐Eun , Alam Md. Jahangir , Kwun In‐Sook 
Published: June 2021

Zinc has been implicated as an activator in the regulation of bone formation. Bone formation is characterized by the synthesis of collagen, the major bone protein in the extracellullar matrix (ECM) which then becomes mineralized by calcium deposition.In this study, we investigated the effect of zinc in the mineralization of two MC3T3‐E1 subclones, SC4 and SC24,cultured in the presence and absence of ascorbic acid. Osteoblastic cells were cultured in normal osteogenic media (OSM), Zn deficient media (1 µM Zn), and Zn adequate media (15 µM Zn) with the addition of 4 µM TPEN as intracellular Zn chelator and supplemented with 75 µg/ml Vitamin C within 20 d of bone formation period.Our results showed that Zn deficiency decreased collagen synthesis and calcium deposition in both subclones. The expressions of ECM bone marker proteins (ALP, OC, OPN) which are necessary for ECM mineral deposition were also lower in zinc‐deficient osteoblasts. Although ascorbic acid is essential in the formation of collagen, Zn deficiency also decreased the synthesis of collagen even in Vit C‐rich media. These results implied that Zn stimulates mineralization by increasing collagen synthesis and calcium deposition which in turn increase expressions of ECM bone marker proteins critical for bone formation in osteoblasts.(KOSEF grant No. R01‐2006‐000‐10640‐0 and KRF grant No. KRF‐2007‐313‐C00810)