bluuebunny/crossref_metadata_2025_split huggingface dataset:
Atomic Structure of Chimeric Hepatitis E Virus Designed for Targeted Nano‐Carrier and Delivery System
Authors: Jariyapong Pitchanee , Xing Li , Weerachatyanukul Wattana , Cheng Holland 
Published: June 2021

The development of either oral vaccines against pathogens or biological container to carry materials for local targeting in digestive mucosa relies on the chief principle in tolerance of vehicle to harsh environment of digestive system to efficiently carry and deliver the cargo to its destination. Delivery system such as enteric‐transmitted virus‐like particles (VLPs) is advantageous because of the inherited acid‐ and proteolysis‐resistance of the parental viral capsids. In the present study, we engineered hepatitis E (HEV) VLP by inserting an antigenic V3 loop of human immunodeficiency virus (HIV) derived peptide, p18, into the recombinant HEV capsid protein (p18‐VLP) after residue tyrosine 485 on the protruding domain (P domain). Using cryo‐transmission electron microscopy (cryo‐TEM) and three dimensional (3D) image reconstruction, quaternary protein structure of the chimeric p18‐VLP reassembled to that of wild type HEV‐VLP with a slight difference at the protruding spike. This engineered particle also reacted to a specific HIV monoclonal antibody, but not HEV antibody at P‐domain suggesting that the inserted p18 was exposed on the surface of VLPs. Even though p18‐VLP was vulnerable to enzymatic cleavage, presumably occurred at the C‐terminal end of the inserted p18 peptide, the p18‐VLP remained structurally intact and maintained particle integrity. The results thus indicate the further application of this HEV‐VLP system as a candidate of oral delivery vector.This research is supported by Commission on Higher Education of ThailandGrant Funding Source: Commission on Higher Education of Thailand
Unraveling the Molecular Mechanism behind Thrombin Induced Myofibrobast Transition
Authors: Meyer Jennifer Ashley , Strande Jennifer 
Published: June 2021

TGF‐β1 is a mediator in fibrosis by activating its downstream Smad signaling pathway, where they are translocated to the nucleus. TGF‐ β has also been shown to inhibit thrombin signaling. Thrombin has been shown to act via the PAR1 receptor, a G‐protein coupled receptor. Activation of PAR1 by thrombin results in many profibrotic effects of thrombin via the induction and activation of secondary mediators including upregulation of TGF‐β and procollagen. Therefore, we hypothesize that thrombin promotes myofibroblast formation through a TGF‐ β independent/dependent pathway.Cells treated with thrombin showed an increase in α‐SMA compared to control. Inhibition of all G‐protein coupled receptors resulted in a complete abolition of fluorescent intensity, indicating that the mechanism by which thrombin induces fibrosis is via G‐proteins, yet inhibition of individual G‐proteins did not have this effect, indicating a potential additive effect. Thrombin treatment also results in increased translocation of Smad2 and 3 to the nucleus. Our results suggest a possible transactivation of the TGF‐ β receptor by the PAR1 receptor. Ongoing experiments will further elucidate the mechanism behind thrombin induced fibrosis.
Comparative analysis of M. pneumoniae models using the CytoSEED plugin for metabolic model visualization
Authors: Hazekamp Nicholas Lee , Kammeraad Joshua , Bockstege Benjamin , DeJongh Matthew 
Published: June 2021

We have developed a Cytoscape plugin for exploring genome‐scale metabolic models in the Model SEED database (www.theseed.org/models). We have used this plugin to compare our metabolic model for Mycoplasma pneumoniae M129 (Mp) with the one published by Yus et al. in Science in 2009. Mp is missing an annotated gene for the transaldolase (TAL) activity, which converts erythrose‐4‐phosphate into sedoheptulose‐7‐phosphate. However, it is known that Mp produces phospho‐ribose‐diphosphate, indicating that the pentose phosphate pathway is functional. The Yus model asserts that the TAL activity is present; however, our software shows that two reactions present in our model associated with fructose‐bisphosphate aldolase and phosphofructokinase bridge this gap using sedoheptulose‐1,7‐bisphosphate as an intermediary compound. Our software also shows differences in asserted transport reactions, glycerophospholipid metabolism, and maintenance of oxidation‐reduction balance. Our software enables comparing the results of flux variability analysis for the two models on different growth media, and the determination of which reactions are essential on the minimal media discovered by Yus et al.NSF 0745100, 0850546
IGFBP‐3 Forms Multimers in Physiologic Conditions
Authors: Mireuta Matei , Hancock Mark , Baxter Robert , Pollak Michael 
Published: June 2021

Insulin‐like growth factors 1 and 2 (IGF‐1 and IGF‐2) are peptide hormones similar in molecular structure to insulin, and they regulate a variety of cellular activities, including metabolism, proliferation, and growth. IGF actions are tightly modulated by a family of proteins called insulin‐like growth factor‐binding proteins (IGFBPs), of which IGFBP‐1 through 6 have been characterized. Most IGFBPs in the blood originate from the liver, but they are also expressed in many other tissues. IGFBP‐3 and ‐5 are the most abundant IGFBPs in the circulation. They form a ternary complex with IGFs and a third protein termed acid‐labile subunit.It has been suggested that IGFBP‐3 and ‐5 can form homomultimers, possibly through disulfide bond exchange, but this effect has been regarded mainly as an artifact. In this work, we show that IGFBP‐3 multimers can occur in physiologic conditions. We demonstrate by surface plasmon resonance that the affinity of the IGFBP‐3 self‐association reaction is in the low nanomolar range. Additionally, we demonstrate by native gel electrophoresis, that IGFBP‐3 multimers of various sizes occur at physiologic concentrations and in several physiologic settings. Moreover, we show that most of these species are SDS‐stable. Our work provides evidence of a previously unrecognized complexity in the IGF‐ 1/IGFBP‐3 system.
AMP‐activated protein kinase (AMPK) activity in Leigh Syndrome French Canadian Type (LSFC) patient fibroblasts: differences between primary and immortalized cell lines
Authors: Mukaneza Yvette , Rivard Marie-Ève , Rosiers Christine Des , Coderre Lise , Consortium LSFC 
Published: June 2021

Background & ObjectiveLSFC is an autosomal recessive disease caused by mutations in the leucine‐rich pentatricopeptide repeat containing protein (LRPPRC) gene resulting in decreased LRPPRC expression. This leads to a tissue‐specific cytochrome c oxidase deficiency and a reduction in energy production. AMPK is a key enzyme in the regulation of energy homeostasis. To test the hypothesis that AMPK activity may differ in LSFC and control cells, we evaluated the phosphorylated and active form of AMPK in primary (P) fibroblasts and, because of the limited availability of P cells, we also tested their immortalized (I) counterparts.ResultsUnder basal conditions, P fibroblasts from LSFC patients and controls showed marginal differences in the levels of phosphorylated AMPK (pAMPK) as well as acetyl‐CoA carboxylase (ACC), one of many AMPK substrates. However, compared to P cells, I cells from both controls and LSFC patients showed a 60% and 80% decrease in pAMPK, respectively. Consistent with pAMPK levels, phosphorylated ACC levels were also decreased in I cells by 58 % in controls and 69% in LSFC patients. The total levels of these proteins were similar in all cases.ConclusionThese results highlight an important decrease in AMPK activity level in I vs. P cells, which may compromise the capacity of these cells to palliate an energy deficit.Supported by: Association de l'acidose lactique and CIHR Emerging Team Grant
Role of cinobufotalin in the pathogenesis of preeclampsia: in vivo and in vitro studies
Authors: Horvat Darijana , Kelso Kelsey , Allen Steven , Fothergill Russell , Zawieja David , Kuehl Thomas , Uddin Mohammad Nasir 
Published: June 2021

ObjectivePreeclampsia (preE) is a hypertensive disorder of pregnancy. Cardiotonic steroids (CS) are endogenous inhibitors of Na+/K+ ATPase. We have shown that one CS, marinobufagenin (MBG), is elevated in preE prior to the development of symptoms. We have also demonstrated that MBG impairs cytotrophoblast (CTB) function, which is critical for placental development. We evaluated the effect of cinobufotalin (CINO), another CS, on pregnant rats and CTB cells to assess its role in preE.MethodsFour groups of rats were studied: normal pregnant (NP, n=10) rats, NP injected with MBG (NPM, n=10) rats, NP injected with CINO (NPC, n=10) rats, and a rat model of preE (PDS, n=10). CTB cell function and signaling were assayed after CINO treatment.ResultsNPC rats showed hypertension and proteinuria similar to PDS and NPM rats. CINO at ≥ 1 nM inhibited CTB cell proliferation, migration, and invasion but had no effect on cell viability. A higher percentage of cells were in the G0/G1 phase in groups treated with ≥ 1 nM CINO. CINO caused an increase in stress signaling p38 MAPK and a positive annexin‐V stain in CTB cells, indicating the activation of apoptotic signaling. However the CINO induced apoptotic signaling was prevented by p38 inhibition.ConclusionsWe have demonstrated that: 1) CINO induces preE in NP rats and 2) CINO impairs CTB cell function. Therefore, CINO may play a role in the pathogenesis of preE.
Analysis of LIPC ‐514C>T and APOA5 ‐1131T>C polymorphisms in Mexican Mestizos and Mexican Amerindians
Authors: Vargas-Martinez Carolina , Contreras Alejandra V , Fernandez-Lopez Juan Carlos , Silva-Zolezzi Irma , Tejero Maria Elizabeth 
Published: June 2021

Description of frequencies and haplotypes in Mexican Mestizo (MM) and Mexican Amerindian (MA) populations, of two SNPs implicated in lipid metabolism, which have been associated to lipid levels in white populations. For this purpose we analyzed DNA extracted from blood samples of 372 MA and 300 MM that participated in the Mexican Genome Diversity Project by allelic discrimination, we also constructed haplotypes and identified tag SNPs.ResultsThe frequency for LIPC rs1800588 was 0.430 in MM and 0.262 in MA, whereas for APOA5 rs662799 the frequency was 0.168 in MM and 0.262 in MA. Three haplotypes (>5%) were identified on the LIPC region on MM, one of them including the rs1800588 (8.3%). Six haplotypes (>5%) were identified on the APOA5 region. The rs662799 SNP was included in haplotypes showing similar frequencies in MM and MA (14% and 14.8% respectively). The APOA5 rs662799 is in high LD with tag SNPs that are present in BUD13 and ZNF259. The LIPC rs1077834, that has been associated to higher high density lipoprotein concentrations in white populations, showed high linkage desequilibrium with our SNP of interest.ConclusionsThe genetic information obtained from this study will be useful in future studies where associations between lipid metabolism phenotypes, environmental factors, and disease onset need to be determine in Mexican population. Source of research support CONACYT.
Mitochondria‐targeted antioxidant therapy reverses age‐related arterial stiffening
Authors: Hearon C.M. , Gioscia-Ryan Rachel A. , Seals Douglas R. , LaRocca Thomas J. 
Published: June 2021

Aging leads to large elastic artery stiffening mediated by increases in superoxide‐dependent oxidative stress and related changes/damage to structural proteins. Mitochondria are a major source of cellular superoxide and may therefore be a therapeutic target for the treatment of arterial stiffening. We studied young (Y: 5 mo, n=5) and old (O: 27–29 mo, n=5) male C57BL6 mice, and O mice treated with the mitochondria targeted antioxidant MitoQ10 (OMQ: 28–29 mo, n=5). Aortic pulse wave velocity (aPWV), an index of arterial stiffness, was higher in O vs. Y mice (485 ± 24 vs. 405 ± 15 cm/s, p<0.05) and was associated with increased aortic superoxide production (electron paramagnetic resonance, O: 7200 ± 1950 vs. Y: 2800 ± 515 AU, p<0.05). O mice also had greater aortic levels of nitrotyrosine (NT), a marker of oxidative stress, and lower levels of manganese (mitochondrial) superoxide dismutase (MnSOD) (western blotting, both p<0.05 vs. Y). MQ treatment (250 μM in water x 3w) normalized aortic stiffening (OMQ: 378 ± 33 cm/s), superoxide production (OMQ: 2835 ± 1131 AU), MnSOD and NT in O mice. MQ also increased expression of complex IV and the mitochondrial biogenesis marker PGC‐1α (both p<0.05 vs. O). Incubation of aortas from Y mice with the mitochondrial superoxide inducer rotenone (0.5 μM, 48h) increased NT by ~300% (p<0.05) and this was inhibited by coincubation with MQ, an effect associated with enhanced MnSOD expression. MQ treatment may be a novel therapy for age‐related stiffening of large elastic arteries by reducing mitochondrial oxidative stress and perhaps increasing mitochondrial biogenesis.NIH AG013038, AG039210
Effect of propranolol treatment on adipocyte lipases and perilipin in severely burned children
Authors: Yfanti Christina , Zhang Chaoying , Collins Sheila , Finnerty Celeste C , Herndon David N , Sidossis Labros S 
Published: June 2021

Severe burn injury is associated with increased peripheral lipolysis, which can be inhibited by administration of propranolol, a non‐selective beta‐adrenoreceptor antagonist. However, the mechanisms responsible for the effect of propranolol on the lipolytic response after severe burn injury are not known.Protein contents of adipose triglyceride lipase (ATGL), hormone‐sensitive lipase (HSL) and perilipin were assessed by western blotting in adipose tissue of 4 patients with severe burn injury and 4 patients with severe burn injury receiving propranolol. Subjects were matched for age (4±3 vs 4±6 years), % total body surface area burn (43±7 vs 49±10), and % 3rd degree burn (40±7 vs 45±6).Propranolol administration for up to 1 week decreased protein content of perilipin by 71% (P=0.06) and Ser‐563‐phosphorylated HSL (pHSL‐563) by 55% (P=0.05). Total HSL content decreased by 32% but did not reach statistical significance (P=0.08). ATGL and Ser‐660‐phosphorylated HSL (pHSL‐660) were not different in the two groups.These preliminary data suggest that propranolol administration acutely regulates specific molecules involved in adipose tissue lipolysis, providing a mechanism for the inhibitory effect of propranolol on adipose tissue lipolysis in severe burn injury.Sources of support: NIDRR (H133A070026 & H133A70019), NIH (P50‐GM60338, R01‐HD049471 & T32‐GM8256), and SHC (84080, 8741, 8660, 9145 & 8760).
Seasonal changes in the hemolymph biochemistry of the snail Helix aspersa
Authors: Bislimi Kemajl Rexhep , Bojniku Hazbije , Gashi Agim , Grapci Linda , Halili Jeton , Mazreku Ilir , Halili Fetah 
Published: June 2021

The seasonal rhythm of the annual concentration of glucose, total proteins, albumins, cholesterol, triglycerides and the activity of aspartate aminotranspherase (AST) and the alanin aminotranspherase (ALT) enzymes in the hemolymph of the snail Helix aspersa (N=180) was researched. The results of the research show seasonal oscillations of the aforementioned parameters in the hemolymph. The highest concentration of the researched biochemical parameters was recorded in the winter, with the significant changes in autumn‐winter (glucose, P<0.001), and summer‐winter (total proteins, albumins, cholesterol and triglycerides (P<0.01). The increased activity of AST and ALT transaminases has been recorded during the summer and early autumn, whereas their lower activity has been recorded in the winter and early spring.The higher concentration of biochemical parameters in the hemolymph during the winter is probably connected with their crioprotector role, whereas the increased activity of the AST and ALT enzymes in the summer could probably be connected to the nutrition and the metabolic level.
Use of Recombinent Casein Peptides as Inhibitors of spPEP
Authors: BASAK SWARNALI , CLACK BEATRICE 
Published: June 2021

Eurygaster integreceps (Sunn pest) causes major damage in wheat grains by injecting their saliva into the grain degrading the gluten proteins. This saliva protein has been identified and characterized as a prolyl endoprotease (spPEP) in our lab (Darkoh et al, 2010). A natural, biological friendly inhibitor to the spPEP is being sought. Previous studies have shown that casein peptides generated by Lactobacillis species have inhibitory effects on mammalian and bacterial prolyl endoproteases (PEP). The Objective of this study was to clone the four bovine casein genes, α1, α2, β and κ for use in expressing each isomer to identify which peptides inhibit the spPEP. The nucleotide sequences corresponding to bovine casein isomers were identified from the NCBI database to design gene specific primers that would amplify the full length casein proteins. Total RNA was extracted from bovine mammary gland biopsy tissue kindly donated by Dr. Erin Connor, Research Leader, USDA‐ARS, Beltsville, MD and subsequent cDNA has been synthesized. Each casein subunit was cloned into the pGEM T‐Easy vector for sequencing. Verified sequences were cloned into the pET15b E.coli expression vector for protein expression and purification using IMAC chromatography. Purified proteins will be used for subsequent incubation with L. helveticus at varying times. The resulting peptide mixture will be incubated with the spPEP in the presence of the GPpNA substrate to determine which isomer/s contains the spPEP inhibitory sequences.Funding Sources: Office of Research and Sponsored Projects, Stephen F. Austin State University, Nacogdoches, TX and Ed and Gwen Cole, Nacogdoches, TX.
Iron stimulates insulin secretion in clonal pancreatic β‐cells and dissociated rat islets
Authors: Erion Karel Arnt , Ferrante Tom , Corkey Barbara , Deeney Jude 
Published: June 2021

Dysregulation of iron homeostasis as observed in people with hemochromatosis and is associated with an increased risk of developing type II diabetes mellitus (T2DM). Also it has recently been shown that dietary iron restriction or iron chelation can prevent the development of diabetes in mouse models of obesity. Our goal was to determine the role of iron in the process of insulin secretion. Clonal pancreatic β‐cells (INS‐1 832/13) and dissociated rat islets were incubated in increasing concentrations of holotransferrin or iron citrate for 1–2 hours. Additionally, perifusions were performed on INS‐1 and whole rat islets. Insulin secretion (HTRF), lipolysis (glycerol release), O2 consumption (seahorse), reactive oxygen species (2′,7′‐Dichlorofluorescein), and PKC activity (MARKS protein) were all measured. Acute exposure of INS‐1 and dissociated rat islets to transferrin bound iron and cell permeable iron salts increases both basal and glucose stimulated insulin secretion (GSIS). This effect is transient and is readily reversible if cells are given a recovery period or are treated with an antioxidant. Acute exposure of both INS‐1 and dissociated rat islets to iron readily increases reactive oxygen species (ROS) which can be blocked using an antioxidant. Iron also increases PKC activity and increases lipolysis in β‐cells. Exposure to iron quickly and reversibly increases both basal and GSIS via a ROS dependent mechanism. Increased PKC activity and lipolysis may play a primary role in this effect.Funding provided by the National Institutes of Health (NIH)
Effects of short‐term protein supplementation on muscle work efficiency in elderly adults
Authors: Sands Kathryn Anne , Clark Kristen L , McKenzie Steven P , Haddad Jeffrey M , Rietdyk Shirley , Campbell Wayne W 
Published: June 2021

ObjectiveThis study assessed the effect of increased dietary protein intake on adaptation of energy expenditure and skeletal muscle function at rest and during low‐intensity physical activity in elderly adults.MethodsUsing a randomized, crossover design, 12 adults (7 F, 5 M), aged 82 ± 7 y (mean ± SD) completed two 6‐day periods of consuming fruit‐smoothie beverages twice daily (with breakfast and lunch) containing 230 kcal and 25 g whey protein (WPro) or 25 g corn‐derived glucose polymer (CHO) (total 460 kcal/day and 50 g/day WPro or CHO).ResultsOn day 7 of each period, fasting blood urea nitrogen was 28 ± 9% higher in WPro vs. CHO (P=0.003), consistent with higher total protein intake. Fasting state energy expenditure and respiratory ratio at rest and while exercising on a stationary cycle at 1, 10, 20 watts were not different between WPro and CHO. Gross mechanical efficiency of skeletal muscle progressively increased from 1 to 10 to 20 watts, but this response was not different between WPro and CHO.ConclusionShort‐term supplementation with either whey protein or glucose polymer does not differentially influence fasting state whole body substrate utilization or skeletal muscle work efficiency in elderly adults.Support: NIH CTSI Grant # TR000006
Association between serum levels of vitamin D with body fat mass, fasting glucose and lipid profile in patients with osteogenesis imperfecta
Authors: Peters Barbara Santarosa Emo , Lazaretti‐Castro Marise , Hayashi Lilian F. , Costa Aline Vasques , Martini Lígia Araújo 
Published: June 2021

AimEvaluate the associations between serum levels of 25(OH)D with body fat mass, glicidic and lipid profile in patients with Osteogenesis Imperfecta (OI).MethodsThirty (30) patients, 16 with OI type I and 14 with OI type III, mean age 26.2 (10.8) years old, were selected at the Bone Fragility outpatient clinic of the Federal University of São Paulo. Weight, height, body mass index (BMI) and fat mass (%FM) (DEXA) were evaluated. Serum levels of 25(OH)D3 were measured by chemiluminescence assay; fasting glucose, total cholesterol, HDL and triglycerides by enzymatic colorimetric assay, and plasma insulin by chemiluminescence detection.ResultsExcess %FM was present in 66.7% of subjects. Only 6.7% of OI patients presented high levels of total cholesterol and triglycerides. All patients presented normal levels of fasting glucose (86.4 ± 8.8 mg/dL) and plasma insulin (7.4 ± 4.9 μUI/mL). The mean serum levels of 25(OH)D3 was 66.5 ± 19.0 nmol/L. No statistical significant difference in biochemical parameters between subjects of OI type I and III were observed. An inverse correlation between serum levels of 25(OH)D3 and fasting glucose was observed (r: −0.652; p:0.03). In OI patients type III there was an inverse correlation between 25(OH)D3 and %FM (r: −0.566; P:0.04).ConclusionThere were associations between 25(OH)D with body fat mass and fasting glucose in patients with OI.Grant Funding Source: Fundação de Amparo à Pesquisa do Estado de São Paulo
Overweight and obesity in children of Arandas, Mexico during the nutrition transition
Authors: Vasquez‐Garibay Edgar Manuel , Rios Lizette Miranda , Velarde Enrique Romero , Cosio Maria Eugenia Nuño , Sanroman Rogelio Troyo , Barrera Liliana Campos , Sabido Erika Caro , Diaz Joanie Ramirez , Tremblay Mark 
Published: June 2021

With the purpose to demonstrate changes on the malnutrition syndrome during nutrition transition a comparison was made in 2000 vs 2010. In the earlier, a cross sectional study carried out in children (60–120 mo) of the Instituto Alteño para el Desarrollo de Jalisco (INADEJ); in the latter a case‐control study with children (60–144 mo) from INADEJ (cases) and children of similar age and sex randomly selected from the education system (Controls). Anthropometric and biochemical indicators, and socio‐demographic variables were included, table 1. Chi square test and odds ratio were estimated.During the last decade, the nutritional status of children living in Arandas, Jalisco are being influenced by socio‐demographic factors of the epidemiological and nutrition transition. This study was sponsored by National Council for Science and Technology, Mexico & Teasdale Corti Grant # 103460–044, Canada.