bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of new cathepsin B inhibitors (585.2)
Authors: Forgas David , Wen Lisa , Huang Jenq‐Kuen , McConnell Rose 
Published: June 2021

Cathepsin B, a cysteine protease that naturally occurs in human cells, is primarily responsible in protein degradation and bone resorption. However, Cathepsin B has been observed in excessive concentrations in patients suffering from certain cancers, as well as Alzheimer's Disease. Though the direct involvement of Cathepsin B within these cells is not currently known, the potential medical benefits of reducing Cathepsin B activities on these cells are promising. The purpose of this research was to evaluate the activity of 5 new synthetic inhibitors of Cathepsin B and to observe the effectiveness of each inhibitor by determining their half maximal inhibitory concentration (IC50). Effectiveness of each inhibitor was determined by the enzyme's ability to cleave the chromogenic substrate, N‐Carbobenzoxy‐L‐Phenylalanyl‐Arginine‐4‐nitroanilide hydrochloride. The activity of the enzyme was observed at 405 nm over a span of 3 minutes. A dose‐response curve was then constructed to interpret the results. The benefits of this research could possibly pave the way for future anti‐cancer treatments, as well as other diseases characterized by over‐expression of cathepsin B.Grant Funding Source: National Cancer Institute at NIH (Grant No. 3R15CA086933‐04 and 3R15CA086933‐04A2S1) and WIU
The calcineurin signaling network evolves via conserved kinase‐phosphatase modules that transcend substrate identity (586.3)
Authors: Cyert Martha , Goldman Aaron , Roy Jagoree , Bodenmiller Bernd , Wanka Stefanie , Landry Christian , Aebersold Ruedi 
Published: June 2021

To define the first functional network for calcineurin, the conserved Ca2+/calmodulin‐regulated phosphatase, we systematically identified its substrates in S. cerevisiae using phosphoproteomics and bioinformatics, followed by co‐purification and dephosphorylation assays. This ‘calcineurinome’ establishes new calcineurin functions and reveals mechanisms that shape calcineurin network evolution. Analyses of closely related yeasts show that many proteins were recently recruited to the calcineurinome by acquiring a calcineurin‐recognition motif. Calcineurin substrates in yeast and mammals are distinct, due to network rewiring, but, surprisingly, are phosphorylated by similar kinases. We postulate that co‐recognition of conserved substrate features, including phosphorylation and docking motifs, preserves calcineurin‐kinase opposition during evolution. One example we document is a composite docking site that confers substrate recognition by both calcineurin and MAPK. We propose that conserved kinase‐phosphatase pairs define the architecture of signaling networks and allow other connections between kinases and phosphatases to develop and establish common regulatory motifs in signaling networks.Grant Funding Source: Supported by NIH 5R01GM048729
Erythrocyte membrane proteins in sickle cell disease (598.1)
Authors: Craik James , Al‐Shammali Hadeel , Adekile Adekunle , Khan Islam 
Published: June 2021

Sickle cell disease (SCD) is a common inherited hematological disorder where mutation causes substitution of valine for glutamate in β‐globin, giving hemoglobin S (HbS). HbS polymerization and cell dehydration cause cell distortion (sickling) at low oxygen tension, often leading to microvascular occlusion. Membrane changes are found in SCD, and increased Na+/H+ exchange activity mediated by NHE‐1 transporter has been reported. This study assayed levels of key nutrient (GLUT‐1 and hENT‐1) and ion (AE‐1 and NHE‐1) transporters to better understand pathophysiological changes in SCD.Membrane protein levels were examined by SDS‐PAGE and quantitative immunoblot analysis (against a common standard arbitrarily set as 100%). Sodium pump (Na/K ATPase) activity was followed by surrogate assay of p‐nitrophenylphosphatase (PNPase) activity.HPLC analysis confirmed HbSS and control phenotypes. SCD showed decreased expression of hENT‐1 (75.33 ± 3.75 n=11 vs. 98.09 ± 7.67 n=11; p < 0.05), elevation of NHE‐1 (108.92 ± 5.01 vs. 93.19 ± 4.90; p < 0.05) and GLUT‐1 (116.92 ± 5.45 vs. 90.94 ± 2.55; p < 0.05) but no significant change in AE‐1 (96.89 ± 2.99 vs. 95.55 ± 2.48) between normal and SCD samples. PNPase activity was increased in SCD (1.303 ± 0. 211 vs. 0.729 ± 0.137 µmole/mg/ml, n= 7; p < 0.05).NHE‐1 increase is consistent with reported Na‐H exchange and decreased hENT‐1 may affect plasma and intracellular purine levels in SCD.Grant Funding Source: Supported by the College of Graduate Studies, Kuwait University, Research Sector Kuwait University R
Accelerated Plb1‐mediated turnover of phosphatidylcholine upon loss of Ypk1 in S. cerevisiae (600.2)
Authors: Surlow Beth , Cooley Ben , Patton Vogt Jana 
Published: June 2021

Ypk1, the yeast homolog of the human serum‐ and glucocorticoid‐induced kinase (Sgk1), is a serine/threonine protein kinase that regulates several aspects of lipid metabolism, including sphingolipid homeostasis and flippase activity. Our results indicate Ypk1 impinges upon the turnover of the major phospholipid, phosphatidylcholine (PC). In pulse‐chase experiments, a ypk1Δ strain displays increased deacylation of PC and the resultant production of glycerophosphocholine (GroPCho) compared to wildtype. Loss of Plb1, a B‐type phospholipase, in the ypk1Δ strain largely abrogates the increase in PC turnover. Furthermore, Plb1 protein and message levels are elevated in a ypk1Δ strain compared to wildtype. A ypk1Δ plb1Δ mutant displays slowed growth and reduced cell wall integrity compared to either of the single deletion strains, suggesting that upregulated Plb1 activity is beneficial in a ypk1Δ backgound. We are in the process of further examining the mechanism by which Ypk1 regulates Plb1. Preliminary data indicate that the regulation of PLB1 expression by Ypk1 is linked to altered sphingolipid metabolism. Our data suggests a novel role for Ypk1 in the regulation of Plb1 mediated PC turnover.Grant Funding Source: Supported by GM NIH R15 GM104876 to JPV
Phosphate starvation in fungi induces the replacement of phosphatidylcholine with the phosphorus‐free betaine‐lipid diacylglyceryl‐N,N,N‐trimethylhomoserine (605.22)
Authors: Riekhof Wayne , Naik Surabhi 
Published: June 2021

Diacylglyceryl‐N,N,N‐trimethylhomoserine (DGTS) is a phosphorus‐free betaine‐lipid analog of phosphatidylcholine (PtdCho) synthesized by many soil bacteria, algae, and non‐vascular plants. Synthesis of DGTS and other phosphorus‐free lipids in bacteria occurs in response to phosphorus (P) deprivation, and results in the replacement of phospholipids by non‐phosphorous lipids. The genes encoding DGTS biosynthetic enzymes have previously been identified and characterized in bacteria and the alga Chlamydomonas reinhardtii. We now report that many fungal genomes, including those of plant and animal pathogens, encode the enzymatic machinery for DGTS biosynthesis, and that fungi synthesize DGTS during P‐limitation. This finding demonstrates that replacement of phospholipids by non‐phosphorous lipids is a strategy used in divergent eukaryotic lineages for the conservation of P under P‐limiting conditions. Mutants of Neurospora crassa were used to show that DGTS synthase encoded by the BTA1 locus is solely responsible for DGTS biosynthesis, and is under the control of the fungal phosphorus‐deprivation regulon, mediated by the NUC‐1/Pho4p transcription factor. Furthermore, we describe the rational re‐engineering of lipid metabolism in the yeast S. cerevisiae such that PtdCho is completely replaced by DGTS, and show that membrane and organelle biogenesis are largely unaffected in the engineered strain.Grant Funding Source: National Institutes of Health
Dynamic interactions of TATA‐box binding protein with promoters is regulated by GlcNAcylation (607.17)
Authors: Hardivillé Stéphan , Han Guanghui , Ma Junfeng , Hart Gerald 
Published: June 2021

The GlcNAcylation is an intracellular posttranslational modification dynamically controlled by two enzymes. OGT catalyzes the transfer of a single O‐GLcNAc moiety from UDP‐GlcNAc to a Ser or Thr residue, whereas OGA removes it. Alterations of the GlcNAcylation profile appear in neurodegenerative disorders, diabetes and cancer. The GlcNAcylation rate of proteins is tightly correlated to the concentration of UDP‐GlcNAc, the end product of the hexosamine biosynthetic pathway, which varies according to environmental and nutritional factors. Interestingly, literature points out a regulation of the basal transcription machinery by O‐GlcNAc. TATA‐box binding protein (TBP) is a key player in regulating transcription since it is required for the activity of all three RNA polymerases. DNA‐bound TBP acts as a dock for recruitment of basal transcription factors and the RNA polymerase II to formation of a functional pre‐initiation complex (PIC). DNA‐bound TBP can also be recognized by NC2 and TAF172. NC2 stabilizes the chromatin interaction of TBP, whereas the association of TBP with TAF172 forms B‐TFIID and promotes its release.Here, we showed that DNA‐bound TBP is modified by O‐GlcNAc on its N‐terminal domain and this GlcNAcylation impairs the recruitment of TAF172 to the DNA‐bound TBP in vitro and the formation of B‐TFIID in vivo. Indeed, using STZ‐treated rat models, we showed that TBP/TAF172 association is decreased in diabetes. Owing the fact that B‐TFIID dynamically regulates the chromatin interaction of TBP, we investigated the dynamic behavior of TBP and TBP O‐GlcNAc by Strip‐FRAP and ChIP assays. Collectively, our results indicate that B‐TFIID formation and TBP dynamics on gene promoters is regulated by O‐GlcNAc.Grant Funding Source: Supported by NIH R01CA42486, R01DK61671, N01‐HV‐00240, P01HL107153
Escherichia coli’s aerobic growth is diminished by small C‐terminal deletions in the regulatory ε subunit of ATP synthase (615.1)
Authors: Shah Naman , Duncan Thomas 
Published: June 2021

FOF1 ATP synthase is a rotary nanomotor involved in energy metabolism. It is also a target of antimicrobial drugs. In bacteria, the ε subunit can inhibit the enzyme with its C‐terminal domain (CTD), which partially enters the rotor cavity to block functional rotation. However, the in vivo role of ε inhibition is unclear since, with the entire εCTD absent, E. coli can grow to nearly normal levels on a non‐fermentable carbon source such as succinate. In the inhibited enzyme, the terminal segment of εCTD is deeply buried in the rotor cavity and interacts with one catalytic subunit. To test the importance of this final segment of εCTD, we genetically truncated the last 5 (εdel5) or last 10 (εdel10) amino acids of εCTD. Growth on succinate was reduced 蠅50% with εdel5 and the effect was similar with εdel10. Thus, perturbing interactions of εCTD with the enzyme is more detrimental to in vivo function than removing the entire εCTD. Initial assays with isolated mutant membranes show i) ATPase activity similar to wild‐type membranes and ii) no dramatic effects on the coupling between the catalytic (F1) and H+‐transporting (FO) complexes. Other in vitro tests are in progress to determine how these truncations diminish the in vivo capacity for respiratory synthesis of ATP. This finding indicates compounds that perturb ε’s regulatory interactions could provide a means to prohibit aerobic growth.Grant Funding Source: Supported by R01GM083088
Fostering remembering: the use of outside of class activities to link learning to prior knowledge (618.14)
Authors: Taylor Ann , Novak Walter 
Published: June 2021

The constructivist model of learning is that concepts are built in the mind of the learner. Biochemistry is a cumulative field, and builds on concepts from general chemistry, organic chemistry, and general biology. The hypothesis of this study is that by correcting errors in these underlying concepts and connecting biochemistry learning to prior concepts will cause better understanding of both the prerequisite knowledge and biochemistry. Students completed activities either targeted at key prerequisite concepts or control activities on a weekly basis. Targeted concepts included equilibrium, bond energy, acid/base chemistry, hydrogen bonding, free energy, stereoisomers, protein function, and alpha helices. Performance was evaluated on a pre/post test of the targeted concepts and on specific questions of a biochemistry final exam.Grant Funding Source: Supported by NSF‐TUES grant #1140711
Evaluating sodium and potassium measuring technologies for human milk (622.1)
Authors: Hartmann Peter , Geddes Donna , Lai Ching Tat 
Published: June 2021

Sodium (Na) and potassium (K) in human milk are associated with the permeability of the lactating breast. We hypothesized that the measurement using an ion selective electrode is similar to inductively coupled plasma atomic emission spectroscopy (ICP‐AES) that requires very low sample volumes.Four groups (low to high fat) of milk samples (n= 30 per group, analyzed in duplicate) were warmed and homogenized. An aliquot (0.3 mL) was used to determine [Na] and [K] by the ion selective electrodes (HORIBA, Japan). Another aliquot (0.01 mL) was digested with nitric acid/hydrogen peroxide and used to determine [Na] and [K] by ICP‐AES (Thermo Scientific, USA). Linear mixed models were used to test whether differences between the two techniques were significantly different from zero for the full data set.Although the measurements using both technologies were within published values, measurement by ICP‐AES for Na and K were (mean mM±SD) 9.8±5.0 and 11.0±4.8,, respectively, which were higher (p<0.05) than by ion selective electrode (5.8±1.5 and 9.1±3.1, respectively). This might due to the complete release of Na and K ions of human milk by the nitric acid/hydrogen peroxide digestion. Therefore, a conversion factor can be applied to the data to allow comparison of values obtained using these two techniques.Grant Funding Source: Medela, AG
Examining participation in school food and nutrition programs among students in grades 6‐8 in Vancouver, Canada (624.23)
Authors: Stephens Teya , Black Jennifer , Chapman Gwen , Velazquez Cayley 
Published: June 2021

Objective: In Canada, both diet‐related health conditions and the environmental sustainability of the food system are growing concerns being addressed by school food and nutrition programs (SFNPs). The objective of this study was to quantify participation in SFNPs among grade 6‐8 students in Vancouver, and whether participation differed by gender, school‐level, or among schools.Methods: A cross‐sectional survey in 26 diverse Vancouver schools assessed student participation in SFNPs (n=937). Rao‐Scott corrected chi‐square tests were used to evaluate associations between participation, gender, and school variables (p < 0.05).Results: SFNP participation varied significantly between schools, yet < 50% of students reported engaging in most SFNPs examined, including: food preparation (36%), choosing or tasting healthy foods (27%), learning about Canada’s Food Guide (CFG) (45%), learning about foods grown in British Columbia (BC) (35%), growing food in a garden (21%), composting (32%), and recycling (51%). Girls were more likely to report learning about CFG or BC foods, and secondary students were more likely to participate in food‐specific activities compared to elementary students.Conclusions: Despite recent efforts to engage Vancouver students in SFNPs, participation in most SFNPs remains low in most schools. Consequently, few students in grades 6‐8 are being exposed to the whole food cycle.Grant Funding Source: Canadian Institutes of Health Research, the UBC Food Nutrition and Health Vitamin Research Fund
Barriers and facilitators to New Mexico Farmers’ Markets’ acceptance of SNAP and WIC benefits (632.21)
Authors: Keane Patricia 
Published: June 2021

Objective: Federal nutrition assistance program benefits allow for purchase of fresh produce at farmers' markets, but the administrative burden on markets is challenging, particularly in small, rural communities. This study evaluates the barriers and facilitators faced by NM farmers' markets in accepting these benefits and suggests policy recommendations and technical assistance approaches to highlight what farmers' markets require in order to fully realize the benefit of participating in these programs, and to ensure low‐income families’ access to fresh fruits and vegetables at farmers’ markets.Methods: Mixed methods Results: Data collection and analysis will be complete at time of presentation.Grant Funding Source: Supported by the University of New Mexico Prevention Research Center
The challenge of classifying foods and beverages marketed to children (632.3)
Authors: Murray Chelsea , Mansuri Sudaba , L'Abbé Mary 
Published: June 2021

Banning the marketing of “unhealthy” foods to children has been proposed as a strategy to address the rising rate of childhood obesity. Criteria have been developed to define what constitutes marketing to children; however, we hypothesize that application of these criteria may yield variable results due to the inherent subjectivity of such criteria. The objective of this study was to evaluate inter‐rater variability by nutrition experts in the classification of products as marketed to children. Eight experts independently evaluated the complete package label of 100 foods from a database of Canadian prepackaged products using the following criteria: 1) allusions to fun/play; 2) child‐oriented lettering/graphics; 3) cartoon/children's characters; 4) toys/prizes/contests/coupons; 5) games; 6) unusual flavour/shape/colour of product; and 7) children's product lines. Classification was inconsistent for 55% of products, with the highest levels of disagreement found for criteria 1) allusions to fun/play and 2) child‐oriented lettering/graphics. The highest agreement among raters was for criteria 4) toys/prizes/contests/coupons and 3) cartoon/children's characters; nevertheless, for these types of product marketing, there was unanimous agreement only 25% and 20% of the time, respectively. This subjectivity illustrates potential challenges in implementing recommendations to ban the marketing of “unhealthy” products to children.Grant Funding Source: Supported by McHenry Chair grant (ML), OGS (CM), CIHR Master's (CM), CIHR PICDP (CM)
Effect of obesity in children on the response to influenza virus vaccination (639.6)
Authors: Rebeles Jennifer , Paulson Justin , Paich Heather , Milner Justin , Xiao Ping‐Jie , Hudgens Michael , Noah Terry , Beck Melinda 
Published: June 2021

Obesity is a global health concern and more than one‐third of the adult population in the United States is obese. Childhood obesity is also on the rise, with obesity rates doubling in children over the past 30 years and tripling in adolescents. Obese adults are at greater risk for increased severity of infections with influenza virus and obesity has recently been found to be an independent risk factor for increased morbidity and mortality from influenza infection. Vaccination with influenza virus vaccine is the best method to protect from or lessen infection with influenza. However, studies in our laboratory have demonstrated that obese adults, when compared with healthy weight adults, have a suboptimal response to influenza vaccination. To expand our findings to children, we enrolled children, ages 2‐17 with a range of BMI percentiles to study the effect of weight status on the response to influenza vaccination. We collected a pre‐vaccination blood sample, and 30 days later, a post‐vaccination blood sample. Peripheral blood mononuclear cells were isolated and stimulated ex vivo with pH1N1 influenza virus, a strain present in the vaccine, and markers of activation and inflammation were measured by flow cytometry. We found that obese children displayed a significant decrease in the CD4+ T cell activation marker CD69, which may suggest that obese children may have a poorer response to influenza infection compared with healthy weight children. In addition, we found that unstimulated T cells from obese children had increased markers of inflammation, including IFNγ and granzyme B, when compared to healthy weight children, suggesting that PBMCs from obese children may not respond appropriately when challenged with antigen. Therefore, much like the obese adult population, obese children also have immune impairments that may put them at greater risk for infections, even when vaccinated.Grant Funding Source: NC TraCS Institute
The effects of a beef‐based meal compared to a calorie matched bean‐based meal on appetite and food intake (640.2)
Authors: Bonnema Angela , Thomas William , Slavin Joanne 
Published: June 2021

Protein and fiber have strong satiety‐inducing potential. Beef is a high quality, protein‐rich food. Beans contain moderate levels of protein as well as fiber. The objective of this study was to determine the effects of a high protein meal (beef) compared to a moderate protein, high fiber meal (beans) on subjective appetite and energy intake at a subsequent meal. Twenty‐eight adults, 14 men (ages 24±5y, BMI 23±2 kg/m2) and 14 women (ages 25±5y, BMI 22±2 kg/m2) consumed two test lunches including a “meatloaf” made from either beef or beans. The beef meal provided 22 grams of protein and 2 grams of fiber while the bean meal provided 14 grams of protein and 10 grams of fiber. An ad libitum snack was given 3 hours after the test meal. Visual analogue scales were used to assess hunger, satiety, fullness, and prospective food intake. Gastrointestinal (GI) tolerance was assessed over 24 hours. No difference between the beef and bean was observed for appetite ratings over 3 hours, food intake at the subsequent meal (632±75 kcal vs 611±75 kcal, respectively), or sum of GI score (2.2±0.5 vs 2.9±0.5, respectively). Gas and bloating were reported more often after the bean meal than the beef meal (2.0±0.4 vs 1.3±0.4, p‐value 0.057). A beef‐based meal with high protein and a bean‐based meal with moderate protein and high fiber produced similar satiety, while the bean‐based meal resulting in higher, yet moderate, gas and bloating.Grant Funding Source: MN Beef Council
Metabolic partitioning of oxidative metabolism in ketotic rat brain (643.2)
Authors: Zhang Yifan , Zhang Shenghui , Xu Kui , Puchowicz Michelle 
Published: June 2021

We have previously reported decreased cerebral metabolic rate of glucose in brain of diet‐induced ketotic rats which inversely correlated with blood ketone levels. These data suggest a “switch” in substrate utilization from glucose to ketone bodies. The metabolic effect of ketosis on brain intermediary metabolism remains to be explored. In this study we investigated the effects of chronic ketosis on partitioning of substrate oxidation. We characterized the 13C fractional contribution of ketones and glucose into citric acid cycle (CAC), GABA and neuro‐amino acids in ketotic rat brain using infusions 13C‐stable isotope tracers. Isoptopomer analysis, gas and liquid chromatography mass spectrometry were applied. Results: 13C percent label contribution from ketone bodies increased 100 fold towards GABA, and 7.9 and 4 fold for glutamate and glutamine with ketosis compared to non‐ketotic rats. The data show chronic ketosis increases ketone body carbon entry towards GABA which is supplied via CAC, suggesting a possible mechanism of neuroprotection.Grant Funding Source: NIH R01 HL092933‐01, R21 NS062048‐01