bluuebunny/crossref_metadata_2025_split huggingface dataset:
structural insights into ERα methylation by histone methyltransferase SMYD2, a cellular event implicated in estrogen signaling regulation (LB267)
Authors: Jiang Yuanyuan , Trescott Laura , Holcomb Joshua , Brunzelle Joseph , Sirinupong Nualpun , Shi Xiaobing , Yang Zhe 
Published: June 2021

Estrogen receptor (ER) signaling plays a pivotal role in many developmental processes and has been implicated in numerous diseases including cancers. We recently showed that direct ERα methylation by the multi‐specificity histone lysine methyltransferase, SET and MYND domain‐containing protein 2 (SMYD2), regulates estrogen signaling through repressing ERα‐dependent transactivation. However, the mechanism controlling the specificity of the SMYD2‐ERα interaction and the structural basis of SMYD2 substrate binding diversity are unknown. Here we present the crystal structure of SMYD2 in complex with a target lysine (Lys266)‐containing ERα peptide. The structure reveals that ERα binds SMYD2 in a U‐shaped conformation with the binding specificity determined mainly by residues C‐terminal to the target lysine. The structure also reveals numerous intrapeptide contacts that ensure shape complementarity between the substrate and the active site of the enzyme, thereby likely serving as an additional structural determinant of substrate specificity. In addition, comparison of the SMYD2‐ERα and SMYD2‐p53 structures provides the first structural insight into the diverse nature of SMYD2 substrate recognition and suggests that the broad specificity of SMYD2 is achieved by multiple molecular mechanisms such as distinct peptide binding modes and the intrinsic dynamics of peptide ligands. Strikingly, a novel potentially SMYD2‐specific polyethylene glycol (PEG) binding site is identified in the C‐terminal domain, implicating possible functions in additional substrate binding or protein‐protein interactions. Our study thus provides the structural basis for the SMYD2‐mediated ERα methylation, and the resulting knowledge of SMYD2 substrate specificity and target‐binding diversity could have important implications in selective drug design against a wide range of ERα‐related diseases.Grant Funding Source: Supported by Leukemia Research Foundation, Aplastic Anemia & MDS International Foundation and American Heart Association.
Activation of caspase‐dependent apoptosis by intracellular delivery of cytochrome c‐based nanoparticles (LB290)
Authors: Morales Moraima , Figueroa Cindy , González Tania , Delgado Yamixa , Molina Anna , Griebenow Kai 
Published: June 2021

Cytochrome c (Cyt c) is an essential mediator of apoptosis when it is released from the mitochondria to the cytoplasm. This process normally takes place in response to DNA damage, but in many cancer cells (i.e., cancer stem cells) it is disabled due to various mechanisms. However, it has been demonstrated that the targeted delivery of Cyt c directly to the cytoplasm of cancer cells selective initiates apoptosis in many cancer cells. Consequently, in this work we designed a nano‐sized targeted Cyt c drug delivery system. First, we precipitated Cyt c with a simple solvent‐displacement method to obtain protein nanoparticles (NPs). Cyt c NPs’ surface was chemically linked to poly(lactic‐co‐glycolic) acid‐ thiol endcap (PLGA‐SH) using succinimidyl 3‐(2‐pyridyldithio)propionate (SPDP) to prevent premature dissolution during delivery. SPDP is a heterobifunctional linker containing a disulfide bond which allows the Cyt c release under intracellular reducing conditions. This construct is further on referred to as PLGA‐S‐S‐Cyt c NPs. The size of the Cyt c NPs obtained was 100‐300 nm in diameter depending on the conditions used, indicating good potential to passively target tumors by the Enhanced Permeability and Retention (EPR) effect. A cell‐free caspase‐3 assay revealed more than 80% of relative caspase activation by Cyt c after the nanoprecipitation and SPDP modification, when compared to native Cyt c. Incubation of HeLa cells with PLGA‐S‐S‐Cyt c NPs showed significant reduction in cell viability after 6 hours. Confocal microscopy confirmed the induction of apoptosis in HeLa cells when they were stained with DAPI and propidium iodide (PI) after incubation with the Cyt c‐based NPs. Our results demonstrate that the PLGA coating, mediated by SPDP linker, stabilizes Cyt c NPs allowing their delivery to the cytoplasm of the target cells. After smart release of Cyt c into the cytoplasm it was able to induce programmed cell death.Grant Funding Source: Supported by the NIH‐RISE Program Grant Number 2R25GM061151‐12
Supplementation of Korean black raspberry improves endogenous antioxidant capacity in overweight adults (LB308)
Authors: Kim You Jin , Lee You Jin , Oh Sang A , Huh Iksoo , Park Saejong , Kim Joohee , Kim Suhkmann , Kim Kyu‐Bong , Park Taesung , Kim Ji Yeon , Kwon Oran 
Published: June 2021

The natural phytochemicals have been well reported as antioxidants. The fruit of Rubus coreanus Miquel (RCM), referred to as the Korean black raspberry, is a rich source of phytochemicals. We hypothesized that response to RCM supplementation would be more sensitive to detect when applied in combination of clinical assessment and nuclear magnetic resonance‐based metabolomics approach. In 4‐week, randomized, double‐blind, placebo‐controlled study, overweight participants (23蠄body mass index (kg/m2)<30) were randomly allocated to receive either RCM or placebo (n=24 per group). At baseline and at the end of the study, each subject underwent an acute bout of treadmill exercise at 60% VO2max for 30 min immediately after ingestion of a single dose of RCM and blood and urine samples were taken at fasting and post‐exercise state. In comparison with placebo, oxidized glutathione, reduced glutathione/oxidized glutathione ratio and superoxide dismutase activity in erythrocyte significantly improved in the RCM group (P‐value<0.05). Urinary metabolites involved in oxidative phosphorylation, amino acid metabolism, glycerophospholipid metabolism and purine metabolism responded significantly to RCM (P‐value<0.05). Taken together, ingestion of RCM may boost the antioxidative defence mechanism and this can be explained by alteration of endogenous metabolites.Grant Funding Source: Supported by the RDA “Cooperative Research Program for Agricultural Science & Technology Development (Project No. PJ0084502013)”, the MSIP through the NRF (Bio‐synergy Research Project NRF2012M3A9C4048761), and the MEST (BK21PLUS 22A20130012143)
A randomized clinical trial to determine the efficacy of manufacturers’ recommended doses of omega‐3 fatty acids from different sources in facilitating cardiovascular disease risk reduction (LB329)
Authors: Laidlaw Maggie , Cockerline Carla , Rowe William 
Published: June 2021

A randomized clinical trial to determine the efficacy of manufacturers’ recommended doses of omega‐3 fatty acids from different sources in facilitating cardiovascular disease risk reduction Maggie Laidlaw1, Carla A. Cockerline1, William J. Rowe:1 1 Nutrasource Diagnostics Inc.Omega‐3 fatty acids confer beneficial health effects, but North Americans are lacking in their dietary omega‐3‐rich intake. Omega‐3 supplementation is an alternative to dietary consumption of fish; however, not all omega‐3 products are created equal. The trial objective was to assess changes in cardiovascular risk following supplementation with four omega‐3 supplements.This was an open‐label, randomized, cross‐over study with thirty‐five healthy subjects. Supplement daily doses (as recommended on product labels) were: Concentrated Triglyceride (rTG) fish oil: EPA 650 mg, DHA 450 mg Ethyl Ester (EE) fish oil: EPA 756 mg, DHA 228 mg Phospholipid (PL) krill oil: EPA 150 mg, DHA 90 mg Triglyceride (TG) salmon oil: EPA 180 mg, DHA 220 mg For the duration of the study, participants were randomly assigned to consume one of the four products, in random order, for a 28‐day period, followed by a 4‐week washout period. This process continued until each subject had consumed each product. Blood samples before and after supplementation were quantified for fatty acid analysis by gas chromatography flame ionization, and statistically analysed using ANOVA for repeated measures.At the prescribed dosage, rTG was statistically superior to all three comparators in the bioavailability of EPA+DHA. Risk reduction in several elements of cardiovascular disease was achieved to a greater extent by rTG than by any other supplement. PL and TG were relatively unsuccessful in this aspect of the study.For the general population, the type and dose of omega‐3 supplements may be immaterial. However, the type and dose of omega‐3 supplementation may be important for those interested in reducing their risk of cardiovascular disease.
A counterfactual analysis of changing economic context and its impact on dietary intakes in Chinese children (LB384)
Authors: Xue Hong , Wu Yang , Wang Youfa 
Published: June 2021

Background: Little is known about how food prices affected children’s nutrient intakes after China’s entry to the World Trade Organization, which has substantial policy implications.Objective: To assess the differential effects of food price change on the distributions of macronutrient intakes in children (6‐18yrs) in Urban China and to simulate potential policy effect.Methods: The China Health and Nutrition Survey (CHNS) data (2000‐2006) with about 1000 children. Quantile regression (QR) and counterfactual analysis were used.Results: Every one Chinese Yuan increase of oil price was associated with about 3 kcal/day (p<0.01) reduction in energy intake from fat for those whose fat intake was below 70 percentile of the fat intake distribution in 2000. In 2006, oil price effect became insignificant. One Chinese Yuan increase in rice price was associated with about 85 kcal/day (p<0.1) energy intake decrease from carbohydrates among those above 90 percentile of the carbohydrate distribution in 2000 with insignificant effect at lower portion of the distribution, which was in contrast with 2006, when rice price effect had a peak impact of 30 kcal/day (p<0.01) at 60 percentile. Protein intake was insensitive to price changes. Counterfactual analysis suggested a 10% increase in rice price would lead to the most effective energy intake reduction ranging from 7‐14 kcal/day.Conclusions: Food prices have differential effects on dietary intakes. Reducing carbohydrate intake could potentially be most effective through pricing instruments.Grant Funding Source: NIH/NICHD (U54 HD070725‐01)
Proteominer facilitates high‐throughput proteomic analysis of low abundance proteins in the cartilage secretome (LB39)
Authors: Williams Adam , Smith Julia , Liddell Susan , Mobasheri Ali 
Published: June 2021

Proteomics is increasingly used for biomarker identification. We have developed high‐throughput mass spectrometric (MS) methods to identify proteins in the secretome of articular cartilage explants, which we are utilizing as an in vitro model of joint inflammation. However, the identification of lower abundance proteins of potentially higher biological significance in this model is hampered by the presence of highly abundant glycoproteins in the secretome. Development of protein depletion techniques that allow the identification of low abundance proteins is important for novel biomarker discovery, especially in the context of identifying pathophysiologically relevant biomarkers of joint inflammation and cartilage degradation. Proteominer protein enrichment kits decrease the dynamic range of complex biological samples, allowing improved analytical coverage of the proteome. The objective of this study was to apply high‐throughput MS techniques following proteominer enrichment to expand the range of proteins that can be identified in the secretome of cartilage exposed to proinflammatory cytokines such as IL‐1β. Explant cultures were established in DMEM + 2% Pen/Strep, using metacarpophalangeal joints of horses. Explants were either untreated controls or treated with 10 ng/ml IL‐1β. After incubating for 6 days at 37⁰C in a humidified environment (95% air, 5% CO2) proteins in culture supernatants were concentrated ~24 fold using Centriplus YM‐3 and Centricon‐3 (3000 MWCO) centrifugal filter units. Concentrated samples were processed with a Small Capacity Proteominer™ Protein Enrichment Kit (Bio‐Rad). After elution, samples were reduced, alkylated, acetone precipitated and trypsin digested overnight, before digested peptides were applied to C18 spin columns, followed by nanoLC‐MS/MS analysis on an amaZon speed ETD. Extracellular matrix proteins identified in untreated control secretome included cartilage oligomeric matrix protein (COMP), fibromodulin and fibronectin. Macrophage migration inhibitory factor (MIF), serum amyloid A protein (SAA) and Heat shock 70 kDa protein 1‐like (HSP70) were identified in explants exposed to IL‐1β. Identification of new low abundance proteins in this model will facilitate a greater understanding of inflammatory processes occurring within cartilage.Grant Funding Source: Supported by Biotechnology and Biological Sciences Research Council (BBSRC) (Contract grant number: BB/G018030/1) and the European Commission Framework 7 program (EU FP7; HEALTH.2012.2.4.5–2, project number 305815).
SDE‐GC‐MS analysis of the Volatile Components in the Wild Grape (Vitis amurensis Rupr) Skin Originated from Zhangjiakou (LB395)
Authors: Zhang Zhisheng , Shu Ying , Li Guijie , Chen Feng , Zhao Congzhi , Tang Xia 
Published: June 2021

In this study, the Wild grape (Vitis amurensis Rupr) which was supplied by a local wine factory of Zhangjiakou City, Hebei Province was analyzed for the volatile compounds in the skin. The volatile flavor compounds were extracted by using the method of simultaneous distillation extraction (SDE) and then analyzed by gas chromatography‐mass spectrometry (GC‐MS) with phenylacetate as the internal standard. Generally the volatile compounds of the wild grape are quite complicated. The analytical results showed that 65 kinds of volatile compounds mostly consisting of 12 esters compounds (2.94 mg/kg), 11 aldehydes compounds (39.96 mg/kg), 14 terpenes compound (4.97 mg/kg), 4 ketone compounds (0.44 mg/kg), 4 phenols compounds (1.33 mg/kg) 7 eneyne compounds (7.69 mg/kg) , 1 alcohol (2.37 mg/kg), 5 heterocycle compounds(1.11 mg/kg) ,5 acid compounds(10.94 mg/kg) and 2 amide compounds (3.47 mg/kg) were detected. The total content of these bioactives is 75.22 mg/kg, in which aldehyde compounds account for the most proportion. Fourteen terpenes were found and the kinds were the most among all of the bioactive compounds; also, they were the main aroma compounds in the wild grape skin.Grant Funding Source: Supported byResearch Center of Primary Product Processing and Engineering of Hebei Province, Baoding
Tolerance and Cognitive Function Effects of a Proprietary Spearmint Extract in Men and Women with Self‐Reported Memory Impairment ‐ A Pilot Study (LB402)
Authors: Nieman Kristin , Herrlinger Kelli , Sanoshy Kristen , Schild Arianne , Kelley Kathleen , Maki Kevin , Ceddia Michael 
Published: June 2021

This pilot trial assessed the tolerance of a proprietary extract from Mentha spicata L. and its effects on cognition in subjects with self‐reported memory impairment. Subjects (N = 11) consumed 900 mg of the extract daily for 30 d. Participants were 73% female and 27% male with a mean age and body mass index of 58.7 y and 27.4 kg/m2. Tolerance was assessed by evaluation of gastrointestinal (GI) tolerability, adverse events, and clinical laboratory tests at baseline and the end of treatment (EOT). Acute cognitive effects were evaluated by comparing pre‐dose vs. 0.5 to 4 h post‐dose values at baseline and EOT. Subjective cognition was also assessed at the EOT. Consumption of the extract for 30 d did not significantly alter GI symptoms and subjective cognition improved (3.5 vs. 4.0; P = 0.063). No serious adverse events or clinically relevant findings in the safety parameters were observed. Cognitive function task scores suggest improved reasoning (24.7 vs. 33.4 points; P = 0.023), attention/concentration (18.0 vs. 40.5 points; P = 0.002), and planning (29.8 vs. 44.1 points; P = 0.088) from baseline after 30 d of treatment. Day one, subjects experienced acutely improved attention/concentration from pre‐ supplementation vs. 2 h (63.4 vs. 82.4 points, P = 0.042 and 18.0 vs. 34.8 points, P = 0.025) and 4 h (63.4 vs. 92.5 points, P = 0.001 and 18.0 vs. 39.8 points, P = 0.002). These results suggest the extract was well‐tolerated and may improve certain aspects of cognitive function. Further investigation in a randomized, controlled trial is underway.Grant Funding Source: Kemin Foods, L.C.
Methylated Medium‐ and Long‐Chain Fatty Acids Provide Novel Sources of Anaplerotic Carbon for Fasting or Exercising Mice (LB438)
Authors: Drayton Josephine , Lin Xi , Jacobi Sheila , Shanely R. , McIntosh Michael , Odle Jack 
Published: June 2021

We hypothesized that methylated fatty acids (e.g. 2‐methylpentanoic acid (2MeP), phytanic or pristanic acids) would provide anaplerotic carbon to the tricarboxylic acid (TCA) cycle and thereby enhance fatty acid oxidation, especially under conditions when TCA cycle intermediates are depleted. The optimal dose of 2MeP and pristanic acid for increasing in vitro [1‐14C]‐oleic acid oxidation in liver or skeletal muscle homogenates from fasted mice (n=4) was assessed using doses of 0, 0.25, 0.5, 0.75, or 1.0 mM. The minimum dose (0.25 mM) maximally stimulated liver tissue oxidation of [1‐14C]‐oleate to 14CO2 by 2‐fold (P < 0.05), but no stimulation was detected in muscle. Similar incubations with 0.25 mM 2MeP, hexanoate, palmitate, phytanic acid, or pristanic acid or 0.1 mM malate or propionyl‐CoA were conducted with liver and skeletal muscle homogenates from acutely exercised or sedentary mice. Regardless of exercise, incubation of liver homogenates with 2MeP increased [1‐14C]‐oleate oxidation to 14CO2 by 60% over control incubations containing hexanoate (P < 0.05). Phytanic acid treatment increased [14C]‐acid‐soluble product accumulation in liver tissue as compared to palmitate (P < 0.05). Anaplerotic stimulation was unaffected by exercised state (P > 0.05) and not evidenced in muscle homogenates. Results were consistent with our hypothesis that methyl‐branched fatty acids may provide anaplerotic carbon and thereby stimulate in vitro fatty acid oxidation.Grant Funding Source: North Carolina Agricultural Research Service and by USDA‐AFRI training grant # 2010‐65200‐20354
Chronic Liver Diseases Which Mallory‐Denk Body (MDB) Form Progress to Hepatocellular Carcinoma is Involved in Ufmylation and FATylation Modification (LB493)
Authors: Liu Hui , Li Jun , French Barbara A , French Samuel W 
Published: June 2021

We previously reported the Mallory‐Denk body (MDB) formation mechanism in DDC induced mice. To further provide clinical evidence whether chronic liver diseases which MDB form progress to hepatocellular carcinoma (HCC) is involved in Ubiquitin‐like proteins (Ubls) modification, gene transcript expression of Ufmylation and FATylation were investigated in human archived formalin‐fixed, paraffin‐embedded (FFPE) liver biopsy and mice frozen liver section from DDC refeding. It showed that all Ufmylation molecules (Ufm1, Uba5, Ufc1, Ufl1 and UfSPs) were at least 50% down regulated both in DDC refed mice and patient liver except for HCC without MDB biopsy, indicating that gene transcript changes was limited in MDB‐forming cells for a signal of proteasomal degredation. Interestedly, FAT10 and subunits of the immunoproteasome (LMP2, LMP7 and MECL‐1) were all up regulated. Among which, a 45‐ and 7‐fold upregulation of FAT10 were observed in DDC refed mice liver and human alcoholic hepatitis with MDB biopsy respectively, implying an important role of this gene. The FAT10‐specific E1 enzyme UBA6, however, was found to down regulation both in patient and DDC refed mice liver. Our results constitute the first systematic demonstration of transcript regulation of Ufmylation and FATylation in liver patient which MDB form progress to HCC and mice MDB‐forming liver by DDC refeding, providing key clue for the investigation of human liver disease development and strategies for drug design.Grant Funding Source: Supported by NIH(RO1 AA021898‐02)
Bordetella pertussis Fimbriae proteins Fim2 and 3 are Immunodominant in T cell responses to Acellular Pertussis Vaccine (LB509)
Authors: Dillon Myles , Kolla Ravi , Sidney John , Sette Alessandro 
Published: June 2021

Objective: To map the response of T cells to the Bordetella pertussis acellular vaccine (aP).Methods: PBMCs from boosted (within three years) and control volunteers were stimulated in vitro with B.pertussis proteins present in the aP. IFNγ and IL‐5 responses were finely mapped using ELISpot assays with a panel of 16‐mer peptides spanning the B.pertussis proteins.Results: Boosted individuals had increased levels of IFNγ and IL‐5 secretion to fimbriae (Fim) 2 and 3, pertactin (PRN), and pertussis toxin (PTx) peptides, while responses to the filamentous hemagluttinin were lower and similar in both groups. The boosting effect induced by aP was most apparent at the level of IL‐5 responses. Within proteins present in the aP, responses to Fim2 and 3 in the boosted group were disproportionately large in relation to the number of peptides tested in comparison to PRN or PTx.Conclusions: The increase in the IL‐5 secretion to Fim2 and 3 reveals that this response is dominant upon aP. Future studies will map the actual epitopes recognized, and compare T cell responses in natural infection to vaccines with and without Fimbriae, in terms of targets of the responses and phenotypes of responding T cells.Grant Funding Source: Supported by: NIH HHSN27220300044C
In silico Assessment of Antihypertensive Potential of Sweet Proteins (LB542)
Authors: Rotimi Solomon , Rotimi Oluwakemi , Omotosho Elizabeth , Iyanda‐Joel Wisdom , Chinedu Shalom 
Published: June 2021

The consumption of low‐calorie artificial sweeteners by patients affected by diseases linked to sugar consumption may be associated with diverse side effects. Hence, sweet proteins derived majorly from under‐utilized plants have been proposed as good replacements. The ability of sweet proteins to release Angiostensin‐Converting Enzyme (ACE)‐inhibitory peptides was investigated. ACE mediates arterial vasoconstriction and elevation of its activity is an important pathogenic mechanism of hypertension. The protein sequences of six sweet proteins, Thaumatin from Thaumatococcus danielli (NCBI accession number, gi|209473), Brazzein from Pentadiplandra brazzeana (NCBI accession number, gi|218218145), Monellin from Dioscoreophyllum cumminsii (NCBI accession number, gi|381144434), Madinlin from Capparis masaikai (NCBI accession number, gi|1817546), Curculin from Curculigo latifolia (NCBI accession number, gi|11225520) and Miraculin from Richadella dulcifica (NCBI accession number, gi|253735645) were selected for sequence alignment using Basic Local Alignment Search Tool (BLAST) analysis and biological activity search using BIOPEP. Although BLAST analysis gave no homologous similarity among the proteins, BIOPEP analysis showed that they demonstrated either di‐ or tri‐peptide with a total of 51, 14, 40, 28, 30 and 59 potential ACE inhibitory peptides from Thaumatin, Brazzein, Monellin, Madinlin, Curculin and Miraculin respectively. The combined digestion with pepsin, trypsin and chymotrypsin A, a simulation of human gastrointestinal digestion released 8, 2, 9, 2, 5 and 11 ACE inhibitory peptides from Thaumatin, Brazzein, Monellin, Madinlin, Curculin and Miraculin respectively. These results add value to these proteins by demonstrating their innate nutraceutical potential in their ability to reduce hypertension.Table of Contents