bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mouse UDP‐glucuronosyltransferase Enzymes Capable of Metabolizing Toxic Estrogen Derivatives: Serine Residues Play Crucial Role to Substrate Turnover.
Authors: RAYCHOUDHURI AMIT , JANA SIRSENDU , BASU MOUSUMI , BASU NIKHIL K , OWENS IDA S 
Published: June 2021

A comprehensive study of mouse UGT isoforms Ugt2b34 and Ugt2b36 was carried in COS‐1 cells in order to understand differences and the significance of apparent adaptations between mouse and human isozymes. Understanding the mouse UGT versus human drug metabolizing enzymes will contribute significant knowledge about our physiology/biochemistry and habitat driven by evolutionary mechanism of dug metabolism. Our study by q‐RT PCR unexpectedly revealed the prostate distribution of these isozymes apart from mouse liver. Glucuronidation assay showed that both Ugt2b34 and Ugt2b36 possess genotoxic 4‐OH estrogen metabolism ability, with Ugt2b36 being far higher turnover number. Ugt2b34 has more substrate range; it is far more effective in metabolizing phyto‐estrogens and phytochemicals. These two enzymes are also effective in metabolizing xenoestrogens like bisphenol A (BPA) and diethylstilbestrol (DES). We also found by site directed mutagenesis that serine‐174 in Ugt2b34 and ser‐172 in Ugt2b36 is essential for the enzymatic activity. Further studies showed that Ugt2b34 is readily inhibited by kinase inhibitors, while, Ugt2b36 is a robust enzyme which is not susceptible to inhibition.This work was supported, in whole or in part, by the NICHD Intramural Research Program.
Protein Scaffolds For Light‐Activated Delivery Of Toxic Iron To Cancer Cells
Authors: Boice Emily N , Kurtz Donald M. 
Published: June 2021

An iron storage protein called bacterioferritin (Bfr) has been adapted for a novel approach to cancer therapy. Tumor‐targeting peptides (TTPs) have been fused to the protein and photosensitizers, like zinc protoporphyrin (ZnPPIX), have been substituted into the native heme binding sites. After binding to cancer cell receptors, photo‐excitation would trigger release of iron to generate a flux of toxic hydroxyl radicals that would overwhelm a cell's defenses.The gene encoding the TTP‐Bfr was synthesized and inserted into an E. coli expression plasmid and was readily expressed with ZnPPIX and purified by standard methods. The protein was then loaded by anaerobic incubation of iron. Bfr solutions were irradiated for various time points. Ferrozine was added to quantitate the iron release. These procedures reproducibly resulted in soluble fully loaded [Fe~1,000ZnPPIX12TTP24‐Bfr]. After 3 hours irradiation, ~30% of the iron was released. At longer irradiation times, 65–70% of the iron was released. No iron was released from dark identical control solutions. We have established that [Fe~1,000ZnPPIX12TTP24‐Bfr] releases significant amounts of iron upon irradiation. Current work is aimed at assessing binding of this protein to melanoma cells, and assessing inhibition of cell growth or apoptosis upon irradiation. Results of these investigations could provide proof of principle for a new approach to cancer therapy.
Stearoyl CoA desaturase 1 (SCD1) and Seipin: Important players in hepatic steatosis?
Authors: Amine Lounis , Lalonde Simon , Rial Sabri , Prieur Xavier , Costet Phillipe , Magré Jocelyne , Mounier Catherine 
Published: June 2021

BackgroundSCD1 is a lipogenic enzyme responsible for the formation of monounsaturated fatty acids, the main precursors of triglycerides. High SCD1 expression is associated with expansion of adipose tissue and ectopic fat accumulation (in various tissues). Seipin is the human Berardinelli‐Seip congenital lipodystrophy 2 gene product which regulates adipogenesis and lipid droplets (LD) formation but whose function remains to be elucidated.AimIn the present study we evaluated the role of SCD1 and seipin in LD formation in hepatic cells and the effect of SCD1 or Seipine deficiency in the lipogenesis and insulin pathways. The results were also validated in seipin‐deficient mice.MethodsIn human hepato‐carcinoma cell line (HepG2), the expression of the two proteins was either silenced. LD formation was monitored by confocal microscopy upon addition of oleate. Under these conditions was also evaluated the activation of insulin and lipogenesis signaling pathways.ResultsWe observed that silencing SCD1 in HepG2 cells decreases the number and the size of LD, does not affect the lipogenic gene expression, increases insulin sensitivity (pAKT and pERK1). While silencing seipin has an opposite effect in the number of LDs. The mice develop a major hepatic steatosis. But the seipin deficiency does not affect lipogenesis but increases the expression of SCD1 and CD36 (in both HepG2 cells and liver of KO mice). Surprisingly, it increases significantly the insulin sensitivity in HepG2 cellsConclusionOur study demonstrated for the first time that low seipin expression in hepatic cells is directly associated with the development of hepatic steatosis. Seipin function appears to be inversely associated with SCD1 function. Finally, our work also suggests that variation of SCD1 and seipin expression influence insulin responses in hepatic cells.
Myristate‐derived d16:0‐sphingolipids constitute an abundant cardiac sphingolipid pool with distinct synthetic routes and functional properties
Authors: Russo Sarah Brice , Tidhar Rotem , Futerman Anthony , Cowart L. Ashley 
Published: June 2021

Previously, our laboratory demonstrated that sphingolipids (SL) mediated diet‐induced diabetic cardiomyopathy in mice (Russo et al. 2012. J Clin Invest). Intriguingly, affected hearts displayed accumulation of a novel pool of myristate‐derived d16:0 SL. However, the metabolism and biological functions of these lipids have yet to be defined. The present study sought to identify the metabolic routes through which these lipids were produced in the heart and to determine whether they were functionally distinct from canonical d18:0 SL. Data revealed that d16:0 SL were produced de novo within cardiomyocytes, comprised a major proportion of the cardiac simple SL pool, and increased in response to excess saturated fatty acids. Additionally, d16:0 and d18:0 SL were shown to be functionally distinct. In isolated adult cardiomyocytes, markers of apoptosis accumulated in response to increased d16:0 SL, but not d18:0 SL. In contrast, accumulation of d18:0 SL, but not d16:0 SL, promoted autophagy in these cells. Thus, these results reveal a previously unappreciated component of the cardiac sphingolipidome with functional differences from canonical sphingolipids.This work was supported by NIDDK F30DK092125 (to S.B.R.), a Merit Award from the Department of Veterans Affairs and the NIH COBRE in Lipidomics and Pathobiology (both to L.A.C.).
The Dynamic Roles of Phospholipid and Triacylglycerol Metabolism in Regulation and Signaling in Yeast
Authors: Henry Susan Armstrong 
Published: June 2021

For several decades, my laboratory has investigated the regulation of lipid metabolism, focusing on how cells sense metabolic activity in the pathways for the synthesis of phospholipids and triacylglycerol (TAG). We isolated mutants with defects in the expression of structural genes of phospholipid biosynthesis, including INO1, encoding inositol 3‐phosphate synthase, the ratelimiting enzyme in the synthesis of the phospholipid precursor, inositol. This led to the discovery of transcription factors and a promoter element responsible for transcriptional regulation in response to inositol. A major breakthrough in the field was the discovery that phosphatidic acid (PA), precursor to both phospholipids and TAG, serves as the signal for derepression of phospholipid biosynthetic genes in the absence of inositol. PA metabolism is influenced by the rates of synthesis and turnover of essentially all lipids that are derived directly or indirectly from it. Because of the interconnectivity of the pathways involving PA, removal or addition of inositol, triggers rapid changes in the levels of a wide spectrum of lipids, including many signaling lipids. We are currently exploiting these attributes of the cellular response to inositol to elucidate the roles of specific lipids, both in regulating lipid metabolism itself, and in activating stress response signaling across multiple membrane compartments.Supported by NIH grant GM‐19629
Characterizing the stabilizing effect of Coq8p and the function of Coq9p in yeast Q biosynthesis
Authors: HE CUIWEN , Xie Letian , Tsui Hui Su , Allan Christopher M , Clarke Catherine F 
Published: June 2021

Coenzyme Q is a mitochondrial lipid that is essential in cellular energy metabolism. Biosynthesis of Q in yeast requires nine proteins, Coq1p‐Coq9p, and deletion of any one of the yeast COQ genes leads to the decreased steady state of several other Coq polypeptides. However, when the Coq8 polypeptide is over‐expressed in some of the yeast coq null mutants, steady state levels of Coq polypeptides were restored to near wild‐type levels. We hypothesize that over‐expression of Coq8p may stabilize the Coq polypeptides by stabilizing the catalytic complex composed of Coq3p‐Coq7p and Coq9p. Using non‐denaturing 2‐dimensional blue native gels, we found that the Coq polypeptide complex dissociated upon the deletion of one of the COQ genes, but that over‐expression of COQ8 rescued the high molecular mass Coq polypeptide complex.The function of Coq9p is still unclear. In the coq9 null yeast mutant overexpressing Coq8 (Coq8 OE), IDMQ6 accumulated when fed para‐aminobenzoic acid (pABA). We hypothesize that Coq9 is required for the deimination step from IDMQ6 to DMQ6. We generated a yeast strain with the COQ9 endogenous promoter replaced by a doxycycline‐ regulated promoter. We found that DMQ6 accumulated earlier and faster than IDMQ6 when there is a very small amount of Coq9p present. The results suggest that the deimination of IDMQ6 is faster than the turnover of DMQ6.
Interleukin‐6 induces ErbBs receptor phosphorylation and utrophin expression in dystrophic myotubes.
Authors: Juretic Nevenka , Díaz Josefina , Jaimovich Enrique , Riveros Nora 
Published: June 2021

Duchenne muscular dystrophy (DMD) is a neuromuscular disease originated by mutations in the dystrophin gene, leading to skeletal muscle fiber loss. A promising pharmacological treatment aims to increase the level of utrophin, a functional dystrophin homolog that could compensate dystrophin absence in DMD muscle fibers. It has been described that interleukin‐6 (IL‐6) and neuregulin‐1 (NRG‐1) induce utrophin expression in skeletal muscle.We investigated a possible functional link among IL‐6, NRG‐1 and utrophin, in normal (C57) and dystrophic (mdx) skeletal muscle cells.Using Western Blot and semiquantitative PCR, we demonstrated that IL‐6 (100 ng/ml) induces NRG‐1 receptor phosphorylation (ErbB2/ErbB3) in both cellular types, and a transient increase on utrophin mRNA levels in dystrophic myotubes. We observed that BAPTA‐AM (an intracellular Ca2+ chelator), GM6001 (a general metalloproteinase inhibitor), genistein (a general protein tyrosine kinase inhibitor) and PD‐158780 (an ErbB receptor tyrosine kinase inhibitor) treatment abolished utrophin mRNA induction.Our results suggest that IL‐6 induces utrophin expression in dystrophic myotubes, through NRG‐1/ErbBs signaling activation, elicited by proteolytic processing of NRG‐1 and ErbBs phosphorylation. Understanding the mechanism involved on utrophin up regulation could help to the development of DMD new therapies.FONDECYT 11100267
Incorporating Biotechnology in the High School Classroom: Professional Development for Biology Teachers
Authors: Bisogno Janet L , Koroly Mary Jo 
Published: June 2021

Advancements in the biological sciences are increasingly dependent upon biotechnology. Although teachers and administrators support the inclusion of biotechnology in the classroom to prepare students for future careers, lack of school funding and minimal teacher exposure to current biotechnology skills have hindered incorporation. The purpose of this project was to develop professional development (PD) sessions for six life science teachers at one high school, utilizing biotechnology equipment and skills developed during programs offered by UF CPET.During the 2011/12 school year, four PD sessions were delivered during regularly scheduled Professional Learning Community (PLC) time. The activities included pipetting, gel electrophoresis, blood typing, and ELISA. School‐based equipment lockers were developed and contained all required equipment and curriculum materials needed for each activity, modeled after the UF CPET locker program. Lockers were available for check out by the teachers, and classroom assistance was available as needed.Evaluations from teachers indicated that they felt more comfortable incorporating biotechnology into their curriculum. Four teachers utilized the pipetting lab and gel electrophoresis lockers, three used the blood typing locker and one used the ELISA locker. For the current year, a fifth PD session and locker are planned to cover microarrays.Sources of research support included ASBMB and UF CPET.
Cutting genes with undergraduate scissors
Authors: Marintcheva Boriana , Young Kelly , Smith Crystal , MacDougall Lawrence , DaConceicao Silviane , Tandon Ankita 
Published: June 2021

Bacteriophage T7 is a genetically and biochemically amenable model system offering great opportunities to train undergraduates in molecular biology and to empower them to pursue independent projects. Students enrolled in BIO497/Undergraduate Biological Research at Bridgewater State University were able to experience science as a process while engineering deletion mutants of T7 DNA metabolism genes.PCR was used to amplify genes of interest from the T7 genome and to clone them using TOPO technology. The resulting plasmids were transformed in E. coli generating strains permissive for growth of phage deletion mutants. In parallel, multi‐step PCR was used to generate recombination cassettes carrying the sequence for cytidine monokinase (cmk), flanked by the natural neighboring sequences of each gene of interest. Cmk is essential for T7 growth, but dispensable for the host, therefore an ideal selection marker for recombinant viruses. In vivo recombination reaction was carried out resulting in the replacement of each individual gene of interest in the viral genome with the cmk sequence.Throughout the experience students learnt multiple techniques, sharpened their problem solving and time management skills, wrote grant proposals, and presented their work locally and nationally. This project was generously supported by the Adrian Tinsley Undergraduate Research Program at Bridgewater State University.
Dietary intake associated with cognitive function of elderly Koreans
Authors: Chung Hae Kyung , Kim Eun‐Ji , Chung Ji‐Hye , Shin Ji‐Eun , Chang Moon‐Jeong , Kim Hye‐Young , Kim Mi‐Hye 
Published: June 2021

Cognitive impairment of the elderly is modulated by nutrients (Vitamin B6, B12, folate and omega‐3), diet (Mediterranean) and nutritional supplement use. We aim to investigate the cognitive function of elderly Koreans in relation to their dietary practices. Elderly Koreans (n=316, 115 men and 201 women), aged ≥65y, were interviewed individually for sociodemographic characteristics, height, weight, FFQ, 24‐hour recalls, and a mini‐mental state examination evaluated by use of a 30‐point questionnaire in Korean (KMMSE). Frequency, Chi‐Square, and Anova tests were carried out using SPSS 12.0. Obesity was more prevalent in men (33%) and those living with spouse and children than women and those living alone. The mean KMMSE scores (± SD) were higher in men (27.55±3.37 vs. 25.53±4.88 in women), the educated, and the relatively younger group (27.77± 2.42 in <70 y vs. 22.67±5.24) in ≥90y) than in their counterparts. Although responses differ by gender, differences in KMMSE scores were associated with frequent consumption of bread, potato, fish, fish cake, pork, beef, radish, tomato, tangerine, apples, grapes, ice cream and rice wine. Intake of vegetables particularly hot peppers, cumbers, tomatoes and seaweed was closely associated with KMMSE scores in Korean elderly.Grant Funding Source: Globalization of Korean Foods Project by Korea Institute of Planning & Evaluation for Technology in Food, Agriculture, Forestry & Fisheries (IPET)
Blood lead, iron deficiency and IQ in Uruguayan first‐graders
Authors: Kordas Katarzyna , Barg Gabriel , Roy Aditi , Manay Nelly , Queirolo Elena I 
Published: June 2021

Cognition in children with both iron deficiency (ID) and elevated blood lead levels (BLL) is not well studied. In 1st‐grade children (n=150, age 6.4±0.6 y, 56.7% boys) from Montevideo, Uruguay we investigated associations among BLL ≥5μg/dL, serum ferritin (SF) <12μg/L), and IQ. Mean IQ, SF and BLL were 92.9±17.0 points, 15.0±14.1 μg/L, 4.7±2.2 μg/dL, respectively. Of the study children, 48.8 and 30.2% had BLL ≥5μg/dL and ID, respectively, 13.4% had both. BLL and SF were not correlated. In unadjusted linear regressions, BLL and SF were not independently associated with IQ; neither were BLL ≥5μg/dL nor ID. However, children with both elevated BLL and ID had lower IQ 11.7 points lower than their ID‐only peers (p=0.07). Preliminary analyses suggest that elevated BLL and ID may result in particularly poor cognitive function in school children.Sponsor: NIEHS 1R21 ES16523–01
Dietary Patterns and prostate cancer risk: a population based cohort study in elderly Swedish men
Authors: Ax Erika Helena , Cederholm Tommy , Grundmark Birgitta , Bill‐Axelson Anna , Becker Wulf , Holmberg Lars , Zethelius Björn , Garmo Hans , Sjögren Per 
Published: June 2021

The aim of this study was to assess the relationship between adherence to a modified Mediterranean Diet Score (mMDS) and a relatively low carbohydrate‐high protein score (LCHP‐score) and prostate cancer (PCa) incidence, in elderly Swedish men. In Uppsala Longitudinal Study of Adult Men, a Swedish population‐based cohort (n=1138, mean age 71±1), diet was determined by 7‐ d food records, and non‐adequate energy reporters were excluded. PCa cases were identified in the Swedish Cancer Register. Risk of PCa was analyzed with Cox proportional hazard regression. Exploratory analyses of nutrients identified by the World Cancer Research Fund as probable modifiers of PCa risk were conducted. During follow‐up (median 13.3y) 72 cases of PCa were identified. The mMDS was not associated with PCa. Adherence to the LCHP‐score was inversely related to PCa, with adjusted hazard ratios (HR) of 0.55 (0.32–0.96) for medium‐, and 0.47 (0.21–1.04) for high‐, compared with low adherence. Inclusion of selenium intake in risk models attenuated the association between the LCHP‐score and PCa, high vs. low adherence HR 0.70 (0.28–1.73).In this study, a moderately low carbohydrate‐high protein dietary pattern was associated with lower PCa incidence. A higher intake of selenium associated with the LCHP‐score was indicated to have contributed to observed relations.
Plain water intake of Korean adults: Korean National Health and Nutrition Examination Surveys 2008–2010
Authors: Kim Jihye , Yang Yoon Jung 
Published: June 2021

ObjectiveThe objective of this study was to examine plain water intake of Korean adults according to anthropometric, dietary, and food consumption characteristics.MethodsThis study was conducted using data from the 2008– 2010 Korean National Health and Nutrition Examination Survey. The subjects were 5,917 men and 8,511 women aged 20–64 years. Water intake was estimated by asking the question “How much water do you usually consume per day?” in the questionnaire. Dietary intake was estimated by 24‐hour dietary recall. A food frequency questionnaire including 63 food items was also administered. Lean body mass and percentage of body fat were measured by dual‐energy X‐ray absorptiometry.ResultsThe mean plain water intake for men and women were 6.3 cup/day and 4.6 cup/day, respectively (1 cup = 200ml). Water intake increased as lean body mass, waist circumference, and body mass index levels increased, except for percentage of body fat. As energy and alcohol intakes increased, water intake increased. However, as energy‐adjusted fat and carbohydrate consumption increased, water intake decreased in men. Water intake increased as energy‐adjusted protein and energy‐adjusted fiber consumption increased in women. As total weight of food intake and total volume of food intake increased, water intake increased. Water intake according to consumption of vegetables was increased in men and women. In the male subjects, water intake increased as frequencies of green tea, alcoholic drink, and all beverages were increased. In the female subject, water intake increased with increased frequencies of green tea, milk, soy milk, and alcoholic drink and decreased frequencies of coffee and soda.ConclusionsOur results suggest that people who had a higher BMI and ingested more food consumed more water. Plain water intake was influenced by the particular types of food ingested such as grains, meat, vegetables, fruits, and beverages.
Validation of a method to determine lutein and lycopene levels in frozen human plasma by reversed‐phase HPLC
Authors: Abadiano Judith , Craft Neal , Yachetti Stephen , Burgher Anita 
Published: June 2021

ObjectiveThis reversed‐phase HPLC method to measure lutein and lycopene in frozen human plasma was validated to support a study of infant plasma from three countries.MethodHuman plasma was maintained at −70 ± 10°C. Unspiked pooled infant plasma provided the Low Quality Control (QC) sample, Mid QC and High QC samples were prepared from adult plasma. Lutein and lycopene in plasma were separated by hexane/tetrahydrofuran extraction then quantified with isocratic reverse‐phase HPLC with visible detection. To validate the method, accuracy, repeatability (n=6) and intermediate precision (n=3) of QC samples were determined.Results

Lutein
Lycopene

NIST n=6
Low QC
Mid QC
High QC
NIST n=6
Low QC
Mid QC
High QC

Mean (mcg/L)
46.4
50
142
292
352
62.3
305
458

RSD* (%)
3.4
5.3
7.2
7.5
2.8
6.4
6.0
8.3

CoA ** (mcg/L)
40 – 54

300 – 380

RSD – Relative Standard Deviation
CoA – Certificate of Analysis

Plasma stability at −70° C for 38 weeks, mean lutein= 95.4% and lycopene=89.1% for the three QC sample pools. The Limit of Quantification (LOQ) for lutein = 5.46 mcg/L and for lycopene = 5.27 mcg/L and defined for this method as the sample concentration that corresponds to the lowest calibration standard.ConclusionThe method has the sensitivity to detect lutein levels from 5.46–292 mcg/L and lycopene levels from 5.27–458 mcg/L in plasma for clinical trials.This is funded by Pfizer, Inc.