bluuebunny/crossref_metadata_2025_split huggingface dataset:
Direct G protein signalling mediates components of the vascular smooth muscle response to insulin‐like growth factor
Authors: Zahradka Peter , Perrault Raissa 
Published: June 2021

Like many organs, the vasculature possesses an intrinsic repair ability that is critical for homeostasis. This ability is intimately linked to smooth muscle cells (SMCs) through their unique phenotypic plasticity, which allows them to proliferate and migrate in response to specific stimuli. However, phenotypic modulation is also the major underlying phenomenon behind pathogenic vascular disease and post‐surgery repairPhysiological control of the SMC response involves the presence of circulating factors, such as Insulin‐like Growth Factor‐ 1 (IGF‐1). IGF‐1 acts via its receptor, a member of the tyrosine kinase receptor family, but it may also require a G protein. Thus, the objective of this study was to investigate the structural aspects of IGR‐1R coupling to a G protein in SMCs, as well as the contribution of this pathway to the cellular response.In primary SMCs, the IGF‐1R was found to interact with both Gαi and Gβ subunits and in the case of Gαi, this was a direct interaction. We found that IGF‐1‐induced activation of ERK1/2 was prevented with a Gβγ inhibitor, which also inhibited cell proliferation in response to IGF‐1. Specific cell‐penetrating Gαi inhibitor peptides were synthesized and these enabled identification of several unique downstream targets.These findings support the conclusion that the responses of vascular SMCs to IGF‐1 are elicited at least in part through interaction with a G protein.Source of research support: Canadian Institutes for Health Research
IGF1‐receptor signalling modulates diastolic cardiac function in aging hearts
Authors: Godecke Axel , Kessels Claudia , Peiseler Lena , Raupach Annika , Jacoby Christoph , Lindecke Antje , Koehrer Karl , Heger Jacqueline , Moellendorf Sarah 
Published: June 2021

Insulin‐like growth factor receptor (IGF1R) signalling appears to play an important role in cardiac growth, contractile function, and the development of age related congestive heart failure. Here we address the question to what extent cardiomyocyte specific IGF1R signalling is essential for maintenance of the structural and functional integrity of the adult murine heart. We inactivated the IGF1R by a 4‐OHTX inducible Cre recombinase in cardiac myocytes of adult mice. Western blots and RT‐PCR demonstrated efficient inactivation of the IGF1R (iCMIGF1RKO)associated with reduced AKT and GSK3β‐phosphorylation. In adult iCMIGF1RKO mice(age 3 month)assessment of cardiac function by P/V catheter and MRI revealed no functional alterations. When the KO was induced in aged mice (11 month) diastolic cardiac function was more depressed in iCMIGF1RKO mice. To test if insulin signalling compensated for the loss of IGF1R, we inactivated β‐cells by STZ. The diabetes associated functional depression was similar in WT and iCMIGF1RKO mice. Analysis of the cardiac transcriptome on 44K microarrays did not reveal activation of overt adaptive processes.ConclusionEndogenous IGF1 receptor signalling is required for conservation of cardiac function of the aging but not the young heart.
Characterization of diverse internal binding sequences to PDZ domains by screening special random peptide library in yeast two‐hybrid system
Authors: Mu Yi , Hu Siqi , Wang Xiaorong , Gao Youhe 
Published: June 2021

There are probably more ways of protein interaction than we currently realize. PDZ domains are flexible protein interaction modules that are considered mainly recognize C‐termini. However, the binding characteristics of PDZ domains to internal sequences have not been systematically studied. In this study, a nearly random octapeptide library with no stop codon designed in the insert was constructed in the yeast two‐hybrid vector. A total of twenty PDZ domains were used as baits to screen the library. They were either previously reported to bind internal sequences or had no known C‐terminal ligand. Among the twenty PDZ domains, eleven were found to bind internal sequences, including three PDZ domains that were not previously reported to bind internal sequences. Eight PDZ domains showed strong amino acids preferences; one of the consensuses was similar to its C‐terminal ligand, but the other consensuses were more diverse than those of common C‐terminal ligands. In addition, one PDZ domain with no strong consensus recognized extended hydrophobic peptides. The results suggest that PDZ domains binding abilities to internal sequences may have been underestimated. Moreover, this data can help us to find potential binding partners which are currently under validation.This work is supported by grants from National Science Fund for Distinguished Young Scholar 30725009.
Large scale analysis of synaptic phosphorylation and O‐GlcNAcylation reveals complex interplay between these post‐translational modifications
Authors: Trinidad Jonathan C , Barkan David T , Gulledge Brittany , Thalhammer Agnes , Sali Andrej , Schoepfer Ralf , Burlingame Alma L 
Published: June 2021

O‐linked N‐acetylglucosamine (O‐GlcNAc) is a dynamic, reversible monosaccharide modifier of serine‐ and threonine‐residues on intracellular protein domains. Crosstalk between O‐GlcNAc and phosphorylation signaling mechanisms has been proposed. However, to date there has not been a large scale evaluation of these two PTMs at endogenous levels in any complex biological system. Here, we identified over 1,750 GlcNAcylation and16,500 phosphorylation sites from the same mouse brain synaptosomes.As has previously been demonstrated for phosphorylation, we show that GlcNAcylation tends to occur in clusters on disordered regions of proteins. However, the distribution of GlcNAcylation sites is virtually random with respect to phosphorylation, suggesting that negative crosstalk at the structural level is not a general phenomenon. We frequently observed proteins extensively phosphorylated with minimal GlcNAcylation. However, proteins extensively GlcNAcylated were almost always phosphorylated to a similar or greater extent, indicating the process of GlcNAcylation appears functionally correlated to phosphorylation.
Unraveling the role of exit tunnel residues of histone deacetylase‐8 in catalysis
Authors: Khanna Ruchi , Singh Raushan K. , mandal Tanmay , Cook Gregory , Srivastava D.K. 
Published: June 2021

Histone Deacetylases (HDACs) are an important class of enzymes which are involved in epigenetic regulations and they have been identified as high priority targets for treatment of cancers. The structural data reveals that apart from the active site pocket, HDAC‐8 harbors an internal cavity through which the acetate product dissociates from the enzyme. To probe the role of the amino acid residues at the “exit port” of the above cavity, we performed site directed mutagenesis of Y18, Y20 and H42 to Ala. Although these mutations did not influence the binding affinity of the substrate, they drastically impaired the catalytic rate of HDAC‐ 8. In case of Y18A mutation, the magnitude of activation by our recently identified activator (TM‐2‐51) was far more pronounced than the wild‐type enzyme. Our circular dichroism studies indicated a change in the secondary structure of the enzyme in case of Y18A mutation (but not in Y20A and H42A mutations) which was manifested in an increased thermal stability of the mutant. Our experimental data reveals that there is a long range communication between the exit tunnel residues and the active site pocket of HDAC‐8, and such communication presumably regulates the overall catalytic feature of the enzymeResearch supported by NIH grants CA113746 and CA132034 to DKS and NIH COBRE grant NCRR‐P20‐RR15566 to GC.
Characterizing the Binding of Enzyme Inhibitors at the Molecular and Ensemble Levels
Authors: Cao Samantha Midung , Pham Benjamin , Radcliffe Amethyst , La Phuc , Wang Richard , An Yi , Sorin Eric J. 
Published: June 2021

Recent studies have shown that butyrylcholinesterase (BuChE) activity is increased in patients suffering from Alzheimer's disease (AD) and is one of the main enzyme targets necessary to treat Alzheimer's symptoms. As BuChE is a key enzyme that catalyzes the hydrolysis of the neurotransmitter acetylcholine, increased control over inhibition of this enzyme is highly sought. We employ molecular dynamics (MD) simulations to study the physics of the inhibitor‐enzyme complex and the chemical characteristics necessary to effectively target BuChE. One family of inhibitors is the dialkyl phenyl phosphates (DAPPs), which have been shown to specifically target BuChE. We studied the interactions and binding affinities of a number of DAPP's docked in the active site of BuChE. Using the Folding@Home Distributed Computing Network, 8,000 all‐atom molecular dynamics simulations were collected, including the enzyme sans inhibitor, which is used to evaluate the mechanism and energetics of ligand‐binding, as well as the equilibrium dynamics of the enzyme‐substrate complex.This work was made possible by the worldwide Folding@Home volunteers who contributed invaluable processor time; Women & Philanthropy, Kenneth L. Marsi, and James L. Jensen Student Scholarships; and a Research Corporation Cottrell College Science Award.
Expression of PPARα, β, and γ in the Hartley guinea pig model of primary osteoarthritis
Authors: El Mansouri Fatima Ezzahra , Nebbaki Salwa Sarah , Zayed Nadia , Benderdour Mohamed , Martel-Pelletier Johanne , Pelletier Jean-Pierre , Fahmi Hassan 
Published: June 2021

BackgroundPeroxisome proliferator‐activated receptors (PPARs) are members of the nuclear receptor superfamily (PPARα, PPARβ/δ and PPARγ). Little is known about the pattern of PPAR expression during the progression of osteoarthritis (OA).ObjectiveTo investigate the expression of PPARα, β, and γ in cartilage over the course of OA in the spontaneous Hartley guinea pig model.MethodsHartley guinea pigs were sacrificed at 2, 4, 8, and 12 month‐old of age. Cartilage was obtained from the central portion of the medial tibial plateau. Cartilage degradation was evaluated histologically using the Osteoarthritis Research Society International (OARSI) guidelines. The expression of PPARα, β and γ was analyzed by immunohistochemistry.ResultsPPARα, β and γ, were detected in medial tibial plateaus from control animals. There was no significant changes in the levels of PPARα and PPARβ over the course of OA. In contrast, PPARγ expression decreased during the progression of OA. Correlation analysis revealed a negative correlation between PPARγ levels and histological score of OA.ConclusionExpression of PPARγ in cartilage decreased during OA. These data suggest that loss of PPARγ expression in cartilage may contribute to the pathogenesis of OA.
Absence of selenoprotein p but not selenocysteine lyase results in neurological dysfunction in mice
Authors: Raman Arjun , Pitts Matthew , Seyedali Ali , Hashimoto Ann , Seale Lucia , Bellinger Frederick , Berry Marla 
Published: June 2021

Selenium (Se) is a trace element that is important for the mammalian central nervous system (CNS). Selenoprotein P (Sepp1) is a Se‐rich protein that is important for brain Se supply, and is thought to supply Se via the enzyme selenocysteine lyase (Scly). Sepp1‐deficient mice develop neurological symptoms of spasticity, and have deficient synaptic plasticity and spatial learning.In this study, male and female transgenic mice lacking Sepp1 or Scly on variable Se diets were evaluated using behavioral and biochemical paradigms for neurological dysfunction. Male mice lacking Sepp1 appear worse than age‐matched female mice, suggesting that males have a higher demand for Sepp1 and Se in the CNS than females. Surprisingly, no behavioral impairment was observed in mice lacking Scly unless they were restricted with a low Se diet. Se deficiency uncovered a mild learning disruption in Scly knockout mice without causing motor deficits. Scly‐deficient mice fed a low Se diet display reduced expression and activity of selenoproteins in brain. These results suggest that Sepp1 is more critical than Scly for CNS function and maintenance of brain Se, but that both gene products are physiologically relevant during dietary Se deficiency.This research was supported by NIH grants RR003061, DK047320, RR016467 and DA027318.
Pannexin 3, a gap junction protein, regulates proliferation through AMPK pathway in odontoblasts
Authors: Iwamoto Tsutomu , Ishikawa Masaki , Yoshizaki Keigo , Nakamura Takashi , Yamada Aya , Yamada Yoshihiko , Fukumoto Satoshi 
Published: June 2021

Pannexin 3 (Panx3), a member of the pannexin family of gap junction proteins, regulates chondrocyte and osteoblast differentiation. However, its expression and physiological functions in tooth development remain unclear. Our objective was to elucidate the expression and identification of molecular mechanisms of Panx3 functions in tooth development. In situ hybridization demonstrated that Panx3 mRNA was preferentially expressed in preodontoblasts. To analyze the role of Panx3 in odontoblast proliferation and differentiation, we used the odontogenic cell line mDP. We found that Panx3 expression was induced in differentiating mDP cells, and overexpression of Panx3 inhibited mDP cell proliferation and promoted cell differentiation into odontoblasts. We also found that Panx3 overexpression promoted the release of ATP into the extracellular space and induced the phosphorylation of AMP activated protein kinase (AMPK), a highly conserved energy sensor in all eukaryotic cells. Further, the activated AMPK pathway inhibited mDP cell proliferation and induced p21 expression. Taken together, our results suggest that Panx3 reduces intracellular ATP levels through a hemichannel, which results in inhibition of odontoblast proliferation though the AMPK/p21 pathway.(Supported by grants from the Ministry of Education, Science, and Culture of Japan and the Intramural Program of the NIDCR, NIH, USA)
CARDIAC MORPHOMETRIC CHANGES IN RENAL ISCHEMIA/REPERFUSION INDUCED CARDIAC HYPERTROPHY IN MICE
Authors: Kmit Fernanda Vieira , Trentin-Sonoda Mayra , Carneiro-Ramos Marcela Sorelli 
Published: June 2021

ObjectivesThe aim of this study was to investigate if particular morphological parameters were altered in heart after renal ischemia/reperfusion.MethodsC57bl/6J mice were subjected to unilateral 60 minutes kidney ischemia, followed by reperfusion of 5, 8, 12, 15 and 20 days. Levels of urea were measured to confirm renal failure. Image captures was taken to analyze morphometric parameters and Cavalieri estimator was performed to estimate heart tissue volume and renal tissue alterations.ResultsUrea levels were increased in groups 5, 15 and 20 days confirming the experimental model. The renal ischemia/reperfusion model promoted a decrease in left kidney weight/ body weight and left kidney length/ tibia length ratio in group 20 days reperfusion as expected. An increase in heart weight and heart weight/tibia length ratio after 15 days of reperfusion was observed compared to sham group (P<0.001). The cardiac hypertrophy was accompanied by a decrease on Left ventricle lumen area in 12, 15 and 20 days when compared to sham (P<0.001), suggesting a concentric hypertrophy. Moreover the ventricles weight showed increased in 15 days group (p<0.001).ConclusionFirst results indicate that renal ischemia/reperfusion model was able to induce several morphometric changes in heart such as concentric cardiac hypertrophy in groups studied.Financial Support: FAPESP and UFABC.
Role of Alpha‐Synuclein and it mutants on Dithiocarbamates Cytotoxicity
Authors: Stoll Anna , Cheng Shu-Yuan 
Published: June 2021

Mn‐containing dithiocarbamates, such as maneb (MB) and mancozeb (MZ), have been known to increase the toxicity of neurotoxin MPTP on dopaminergic neurons leading to cell death and Parkinson‐like symptoms. Dopamine transporter (DAT), a key protein in MPTP's toxicity, transports the active metabolite (MPP+) into dopaminergic neurons; increasing the cell surface expression of DAT increases uptake of MPTP. Alpha‐synuclein (both wt and A30P & A53T mutants) have been associated with Parkinson's, and are thought to regulate DAT trafficking by interacting with DAT.The aim of this study is elucidate the role of the alpha‐synuclein and its mutants on dithiocarbamate cytotoxicity. HEK cells transfected with DAT and alpha‐synuclein (either wt or mutant) were treated with dithiocarbamates. Lysates were used for co‐immunoprecipitation using anti‐DAT antibody. The precipitated proteins were subjected to Western blot analysis using alpha‐synuclein and DAT antibodies.We report roughly a 30% decrease in the DAT/alpha‐synuclein interaction in the presence of the A53T mutant versus wild‐type, but not in the present of A30P mutant. With dithiocarbamates the interaction of DAT and alpha‐synuclein (wt) was enhanced, with increases ranging from 50% to 200%. Pesticide treated A53T mutants respond more dramatically than A30P mutants in increasing the interaction between the DAT and the alpha‐synuclein.
Regional anatomy of the prostate: Are we providing the most clinically relevant material during anatomy courses
Authors: Matsler Nik , Benninger Brion 
Published: June 2021

INTRODUCTIONProstate pathology is inevitable for the aging male (50% by 60, 90% by 80). Despite this, students are not traditionally required to dissect the prostate and the intimately associated structures. The objective of this study was to investigate clinically relevant prostate anatomy in a dissection course.METHODSLiterature search of contemporary anatomy textbooks, atlases, websites, and specialty journals was conducted regarding the prostate and associated anatomy. Urology surgeons were consulted for relevant anatomy. Dissection of the prostate region on embalmed male cadavers was conducted.RESULTSLiterature search revealed a difference in emphasized clinically relevant structures between urologists (specialty journals) and anatomy texts/atlases. Cadaver dissection revealed successful identification of clinically relevant structures.DISCUSSIONProstate signs and symptoms are clinically significant and prevalent in the population. Surgery is a common treatment of benign and malignant prostatic hypertrophy. Dr. P.C. Walsh recognized the benefit of detailed anatomical knowledge while developing a neurovascular preserving technique, which reduced postoperative symptoms and side effects.CONCLUSIONThis study suggests that contemporary anatomy texts and atlases could add further emphasis to clinically relevant structures of the prostatic region.
Dietary Advanced Glycation End‐products (AGEs) of Patients with Diabetic Foot Ulcers (DFU)
Authors: Maier Haiyan , Spicer Maria 
Published: June 2021

Dietary AGEs, also known as glycotoxin, is associated with inflammation and oxidative stress in Diabetes Metillus (DM). Exogenous AGEs in food are the major contributors to total body AGEs. The objective of this study is to determine whether AGEs intake and other dietary factors influence wound healing duration and wound severity of patients with DFUThe research protocol was approved by Human Subjects Committee of The Florida State University. Thirty DFU patients between 45–75 years of age were recruited from wound care centers in Tallahassee, FL. Three 24 hour food recalls with emphasis on cooking methods were collected from each participant. ESHA Food Processor SQL was used for food intake analysis. AGEs content was determined based on a published database by Goldberg (2004). Statistical analyses, include correlation of AGEs and wound healing duration, were performed using IBM SPSS Statistics 19.Initial analysis of the data indicates a positive trend for wound healing duration and dietary glycotoxin, as well as saturated fat intake. Patients with DFU have higher fat intake, especially saturated fat, and, lower fruit and vegetable intake compared with the U.S. dietary guidelines.Decreasing dietary glycotoxin and fat intake may promote faster wound healing and may be used as a predictor for DFU healing time.
Polyunsaturated fatty acids levels in toddlers are related to maternal FADS2 DNA methylation and rs174575 genotype
Authors: LUPU DANIEL , NICULESCU MIHAI D. , CHEATHAM CAROL L. , STEGALL JUDITH M. 
Published: June 2021

DL and MDN contributed equally to the scientific content of this abstract.The aim of this study was to evaluate the influence of maternal genotype and DNA methylation status of the fatty acid desaturase 2 (FADS2) upon plasma PUFAs concentrations in toddlers.108 toddlers (16 months old) and their mothers were enrolled in a double‐blind intervention study (see abstract no. 6386). At the study start, maternal and toddlers’ FADS2 genotype (rs174575), maternal FADS2 promoter DNA methylation, and toddlers’ plasma fatty acid profiles were assessed using real time PCR, bisulfite pyrosequencing, and capillary gas chromatography, respectively. Using linear regression we identified significant correlations between maternal genotypes/FADS2 promoter methylation and omega‐3 and 6 species. In addition, we have identified one CpG site within maternal FADS2 promoter, with the highest correlation to α‐linolenic acid (18:3, ALA) plasma concentrations in toddlers. Stratification by gender indicated that such correlations are stronger for male toddlers as compared to females. Pending ongoing determinations in toddlers, our preliminary conclusion is that maternal FADS2 genotype (rs174575) and DNA methylation status had in imprinting effect upon the metabolism of omega‐3 and 6 fatty acids in toddlers. Investigations are ongoing to determine whether imprinting is of metabolic or epigenetic nature.
Conceptualization and Assessment of Nutrition Needs in Intermarried Couples in South Korea
Authors: Hwang Ji-Yun , Kim Eunjung 
Published: June 2021

No attempt was made to understand nutrition barriers for intermarried couples in South Korea. This study was undertaken to understand how Vietnamese marriage immigrants and Korean husbands perceive and assess their need for nutrition education and program. Qualitative in‐depth interviews were done with a sample of Vietnamese marriage immigrant women and Korean spouses participating in or having experienced Nutrition Plus from Community Health Centers in Seoul, Korea. Each interview was recorded using digital voice recorders, transcribed, and analyzed. Perceptions of nutrition needs were assessed by comprehensive understanding of the extent to which an intermarried wife or a Korean husband has barriers of eating properly to maintain good nutritional status. Qualitative analysis revealed that need was determined by various cultural, economic, and environmental factors including food insecurity of the intermarried households, rather than by a single characteristic of them. Suggestions are presented regarding future efforts to better understand and meet the nutrition needs of intermarried couples to respond to their heterogeneous needs and to deliver adequate nutrition service to ever increasing intermarried families.This study was supported by the grant from National Research Foundation of Korea (2011‐0014986).