bluuebunny/crossref_metadata_2025_split huggingface dataset:
Exploring the Structural Link Between FOXP2 and ADHD in Human Adults
Authors: Savoy Kaitlyn , Nataraj Kruti , Olvera Sophia , Petursson Emma 
Published: April 2020

The forkhead box protein P2 (FOXP2) gene, located on chromosome 7, codes for a forkhead/winged‐helix transcription factor which regulates transcript production, typically repressing expression of neutral targets. The Olathe North MSOE Center for BioMolecular Modeling SMART Team used 3‐D modeling and printing technology to examine the structure‐function relationship between this protein, FOXP2, and attention deficit hyperactivity disorder (ADHD) in human adults. An assessment of the relationship between the presence of each of 12 FOXP2 tagSNPs and the presence of ADHD in two European samples found 1 tagSNP to be significantly associated with combined ADHD in one sample in both a single‐ and multiple‐marker test. These encouraging results provided the foundation for further exploration of the structural link between FOXP2 and ADHD in adults. Current studies involve observation of a variety of samples and deep‐sequencing of the FOXP2 genomic region with the intent to identify the sequence variants directly involved in the genetic background of ADHD. Future studies may focus on genes regulating and regulated by FOXP2 to gain a better understanding of the mechanisms and functions of the protein.Support or Funding InformationThe Olathe North MSOE Center for BioMolecular Modeling SMART Team used 3‐D modeling and printing technology to examine the structure‐function relationship between this protein, FOXP2, and ADHD in human adults.Sponsors: Chris Elniff and L.B. FogtMentor: Brendan Mattingly, University of Kansas Edwards
Adiponectin Reduces Angiotensin II‐Induced Vascular Hypertrophy by Targeting ROS Formation and Actin Cytoskeleton Remodeling
Authors: Zeidan Asad , Radwan Zeina , Farht Rima , Ghantous Crystal 
Published: April 2020

Background and aimsAdiponectin (APN), an adipocytokine, exerts cardioprotective effects, while angiotensin II (Ang II) plays a critical role in the pathogenesis of hypertrophy of vascular smooth muscle cells (VSMCs) induces hypertension and vascular remodeling. Ang II induces VSMC hypertrophy in part via activation of RhoA/ROCK pathway. In this study, we investigated the molecular mechanisms associated with Ang II‐induced RhoA activation and translocation and the ability of ANP to counteract Ang II‐hypertrophic effect through inhibition of RhoA activation, actin cytoskeleton remolding and reactive oxygen species (ROS) formation.MethodsStudies were carried out using rat aortic tissues and VSMCs. The effect of APN on Ang II‐induced RhoA translocation was assessed by Western blotting (WB) and Immunohistochemistry. Leucine incorporation was used to examine NO effect on Ang II induced hypertrophy. The phosphorylation of cofilin‐2 and ERK1/2 was detected by WB. Moreover, immunohistochemistry was performed on aortic frozen sections to detect ROS, F‐actin and G‐actin levels, RhoA‐Cav 1 co‐localization and eNOS translocation.ResultsAng II‐induced VSMCs hypertrophy was prevented by supplementing APN. The hypertrophic response to Ang II was associated with RhoA translocation to caveolae‐rich membrane, cofilin‐2 and ERK1/2 phosphorylation, in addition to a significant decrease in G/F actin ratio. Moreover, ROS production significantly increased with Ang II treatment as revealed by confocal microscopy. However, these effects were inhibited by APN and caveolae disruption. Taken together, these results suggest that Ang II hypertrophic effect is due, in part, to RhoA translocation to caveolae rich micro‐domains and its downstream effectors which is counteracted by APN.ConclusionThese findings suggest the ability of APN to attenuate Angiotensin II‐induced vascular hypertrophy.
Formalism of Euler’s Graph Theory is Suitable for Natural Etiopathogenetic Pathways and Clusters Mapping ‐ Integrative Study of Pathophysiology
Authors: Kovac Zdenko 
Published: April 2020

Leonard Euler (1707–1783) introduced the graph theory, a special part of geometry, as a tool to “… deal with relation dependent on position alone, and investigates properties of the position; it does not take magnitude into consideration, nor it does involve calculation …”. Graph theory is useful concept for nonlinear complex systems. As non‐numerical and non‐metric mathematical reasoning, it describes a complex system as a net of multiple events, seen as a whole. Similarly, graphic representation of physiological networks has an advantage in comparison with traditional narrative descriptions.We invented matrix guided analysis and re‐synthesis method to outline and visualize etiopathogenetic pathways and their natural interconnectivity. It is four steps method (consisting of a patient problem exposition, repetition of relevant knowledge, a design of algorithm and systematization) that enforces the horizontal, vertical and longitudinal integration of knowledge. The algorithmic pathways and networking of processes are crystallized out of plethora of clinical data (signs, symptoms, dysfunctions, outcomes, etc) and knowledge of sub‐systems (nano‐molecular, thermal, macromolecular, cellular phenomena, etc.), which are largely acquired via reductionistic methodologies.Algorithmic pathophysiology elaboration has revealed a natural tendency to form common crossing points of reactivity of heterogeneous pathways. The pathways spontaneously converge and form etiopathogenetic clusters (EPCs). EPCs integrate multiple inputs and multiple exits. They look like natural integrators, the common “hubs” of human body response in various diseases. 91 EPCs have been identified at multiple levels of hierarchy of human body disease reactivity. EPCs are shared in natural development of heterogeneous conditions and they show tendency to form a network. Among the EPCs connecting pathways serve as “the EPC‐driving force”. Thus, the EPCs may be seen as “a new emerging feature” of pathophysiology foundation of disease.Following Euler’s strategy directed links in‐between elements of algorithm indicate a “physiological flux”, mutual oriented influence, and/or cause‐consequence. Physiological relations among acting elements have directionality of action, feedback and feed‐forward regulatory loops. Graphic maps outline the parallel pathways and easily put together relevant events of nano‐scale analyses with the clinical macro‐scale level. The (EPCs) are crossing and integrating points of complexity networks. The network visualization of physiology of disease and health underlying processes places individual mechanisms within the complex interconnectivity and may contribute to explanation of nonlinearity phenomena.
Amino Acid Substitution at Phe80 of Mammalian Phenylalanine Hydroxylase Destabilizes Both Resting‐State and Activated Conformations Increasing the Population of Intermediates.
Authors: Arturo Emilia C. , Merkel George W. , Borne Elias , Hansen Michael Riis , Lisowski Sophia , Gupta Kushol , Jaffe Eileen K. 
Published: April 2020

Phenylalanine hydroxylase (PAH) is an allosteric enzyme responsible for maintaining phenylalanine (Phe) below neurotoxic levels; its failure results in phenylketonuria (PKU). PAH equilibrates among long‐lived conformations, including resting‐state (RS‐PAH) and activated (A‐PAH), whose equilibrium position depends upon allosteric Phe binding to the A‐PAH conformation. The RS‐PAH conformation contains a stabilizing cation‐pi sandwich between Phe80, Arg123, and Arg240 (PDB entry 5DEN), which cannot exist in the A‐PAH conformation. Intrinsic protein fluorescence, enzyme kinetic analysis, native PAGE, size exclusion chromatography, limited proteolysis, and behavior on ion exchange resin are reported for F80A, F80D, F80L, and F80R, many as a function of [Phe]. These data indicate that amino acid substitutions at Phe80 destabilizes both the RS‐PAH and A‐PAH conformations so that intermediate, on‐pathway conformations are longer lived. The addition of Phe allows stabilization of the A‐PAH conformation. Kinetic characterization of F80A and F80D reflects allosteric activation while F80L and F80R are constitutively active. The reaction rates of all Phe80 variants suggest relief of a rate determining conformational change present in the wild type protein. Limited proteolysis of WT rPAH in the absence of Phe reveals facile cleavage within a central C‐terminal 4‐helix bundle, reflecting dynamic dissociation of the PAH tetramer. Under these conditions, the Phe80 variants show proteolytic hypersensitity in a linker region that repositions in the RS‐PAH to A‐PAH conformational interchange; this is protected by addition of Phe. We conclude that manipulation of Phe80 dramatically affects the conformational space sampled by PAH, increasing the population of intermediates between RS‐PAH and A‐PAH.Support or Funding InformationNIH 5R01‐NS100081; NIH P30 CA006927
Advanced Glycation End Products Downregulate Placental Growth Factor Expression in Endothelial Cells Via a Macrophage‐Mediated Mechanism
Authors: Semola Yemi , Lovern Pamela 
Published: April 2020

In coronary artery disease, atherosclerosis chronically reduces blood flow. Placental growth factor (PLGF), a protein produced by endothelial cells (EC), can promote collateral artery remodeling to increase blood flow. However, collateral growth is impaired in people with type II diabetes. Our group has previously demonstrated that PLGF levels are reduced in a mouse model of diet‐induced type II diabetes. However, the mechanism for this reduction remains to be defined. Inflammatory cytokines and advanced glycation end products (AGE) are implicated in diabetic vascular dysfunction. We therefore hypothesized that AGE and TNFα signaling would inhibit PLGF expression in EC. Human coronary artery EC were treated with AGE, bovine serum albumin (BSA, as a control for AGE), or TNFα. AGE had no direct effect on PLGF in EC; however, TNFα reduced PLGF in EC (P<0.001). Next, murine macrophages (RAW cell line) were treated with AGE or BSA. RAW media was then transferred to murine endothelial cells (EOMA cell line). Treatment with both RAW‐BSA and RAW‐AGE media significantly reduced PLGF levels in EOMA cells compared with control RAW media (BSA, 481.75 ±21.00 pg/mL; AGE, 631.29 ±41.88 pg/mL; control, 1022.48 ±115 pg/mL; P<0.001, n=4). However, TNFα levels were only increased in RAW‐AGE media compared to RAW‐BSA media and RAW‐control media (AGE, 811.11 ±48.37 pg/mL; BSA, 305.68 ±13.95 pg/mL; control, 276.07 ± 10.40 pg/mL; P< 0.001, n=3). This finding suggests that cytokines other than TNFα may also be inhibitory for PLGF. Analysis of RAW media using a Multiplex Pro mouse 6‐panel cytokine assay revealed that TNFα was increased in RAW‐AGE media compared to RAW‐BSA and RAW‐control media (P<0.007). There was also a trend towards increased IL‐10 and IFN‐γ with AGE treatment, which approached significance (P<0.06), suggesting that these cytokines may also be important targets for future investigation. Identifying signaling pathways by which AGE and inflammatory mediators affect PLGF will give new insights into the pathology of diabetic cardiovascular disease.Support or Funding InformationNIH R56 HL084494 (PL), OSU CVM
Substrate Binding to the Catalytic Domains of Lysine Deacetylase 6
Authors: Eyassu Daniel G. , Williams Ayanna J. , Toro Tasha B. , Watt Terry J. 
Published: April 2020

Thousands of acetylated protein sequences have been identified in humans. Lysine deacetylases (KDACs or HDACs) are enzymes known to reverse acetylation by catalyzing hydrolysis of acetyllysine residues. Proper regulation of acetylation levels by KDACs and the opposing lysine acetyltransferases plays a role in many biological processes. Irregular activities of KDACs have been associated with a variety of diseases including various cancers. KDAC6 is the only known human KDAC to have two catalytic domains (CDs). The individual contributions of the catalytic domains to the overall activity of the enzyme and interaction with substrates are not well understood. Additionally, there are not many known substrates for the enzyme. We hypothesized that each CD has independent deacetylase activity and different substrate selectivity. To test this hypothesis, the activity of each domain was determined with a variety of peptides, many derived from acetylated proteins that are putative substrates of KDAC6. The second catalytic domain appears to be responsible for the majority of catalysis. To determine why peptides were active with one domain and not the other, we used molecular dynamics simulations to model the interaction between the CDs and the peptides. We identified residues relevant in interactions between each catalytic domain and peptide pair. Activity data and apparent substrate affinity were used to validate molecular dynamics results as explanatory for differences in substrate affinity for each CD.Support or Funding InformationNIH TL4GM118968, UL1GM118967, 5G12MD007595, and R15GM129682; NSF CHE 1625993 and MCB 1817358; U. S. Army Research Laboratory and the U. S. Army Research Office W911NF1810450; and the Louisiana Cancer Research Consortium.
Effects of prebiotic inulin pre‐treatment on susceptibility to DSS‐induced colitis in mice
Authors: Nadolski Nolan L.W. , Prawitz Rachael , Winker Olivia , Walton Kristen L.W. 
Published: April 2020

Inflammatory bowel disease (IBD) is a chronic intestinal disease that includes Crohn’s disease and ulcerative colitis. Previous studies have found alterations in the microbiome in IBD and in animal models of IBD, underscoring the role of the intestinal microbiota in IBD pathogenesis. Prebiotics are molecules in foods that support the growth or activity of beneficial microbes. Inulin, a soluble fiber molecule, is a commonly available prebiotic, and previous studies suggest that it can reduce symptoms in dextran sodium sulfate (DSS)‐induced colitis in mice. This study aimed to determine whether pretreatment with 1% inulin for 1 week, followed by 7 days of 3% DSS administration with continued inulin treatment, altered the severity of disease in C57Bl/6 mice. Clinical signs of colitis, including weight loss, fecal occult blood, and fecal consistency, were monitored daily during the DSS treatment period. At the end of the experiment, samples were collected for histological scoring, and for RNA isolation and analysis of colonic pro‐inflammatory cytokine expression by quantitative PCR. Unexpectedly, inulin pre‐treated, DSS‐exposed mice were positive for blood in the stool a day sooner than mice given DSS only. However, by the end of DSS treatment, the disease activity index was not different between the two groups. In addition, colon length and colon wet weight were similar in inulin‐treated mice and non‐inulin treated mice. Histological scoring of colitis showed no difference between inulin‐treated and non‐inulin‐treated colitic mice. In conclusion, 7 days of pre‐treatment with inulin followed by induction of acute DSS‐induced colitis did not significantly alter susceptibility to colitis.Support or Funding InformationSupported by the Missouri Western State University PORTAL program
Expression and Protective Functions of the sst4 Somatostatin Receptor in the Indomethacin‐Induced Gastrointestinal Mucosal Injury Model of the Mouse
Authors: Helyes Zsuzsanna , Csekoe Kata , Kormos Viktoria , Zelena Dora , Iarushkina Natalia , Sudalina Maria , Horvath Adam , Kecskes Angela , Gaszner Balazs , Filaretova Ludmila 
Published: April 2020

IntroductionGastrointestinal ulceration is associated with high mortality despite advanced management, therefore, understanding of the mucosal protective mechanisms is important. Somatostatin is present in the endocrine cells and sensory nerves, and its analogues were shown to reduce mucosal blood flow, pepsin and gastric acid secretion. Our group discovered that its anti‐inflammatory and analgesic actions are exerted via the somatostatin receptor subtype 4 (sst4). Therefore, we investigated its expression and role in gastrointestinal injury.MethodsSst4 gene‐deleted (sst4−/−) and wildtype mice were treated s.c. with indomethacine (IDM; 35 mg/kg) or vehicle. RNA scope in situ hybridization was performed to localize, qPCR to quantify sst4 and somatostatin mRNA. Macroscopic and microscopic lesions in the stomach and small intestine were evaluated after 4 and 48 h. Plasma protein extravasation was measured by fluorescent, neutrophil myeloperoxidase (MPO) activity bioluminescent ex vivo imaging, TNFα, IL‐1 and COX2 mRNA by qPCR.ResultsSomatostatin mRNA is present in the neuroendocrine D‐cells and significantly upregulated 48 h after IDM. Sst4 mRNA in located in the myenteric plexus neurons, it’s expression did not change in response to mucosal injury.IDM‐induced gastric and small intestinal macroscopic lesions (4, 24 h), gastric MPO activity (24, 48 h) and plasma extravasation (30 min) were significantly greater in sst4−/− mice. TNFα, IL‐1, COX2 expressions significantly increased after 4 h in the stomach of sst4−/− mice, but in wild‐types the same pattern was observed only later, at the 48 h time‐point. IDM‐evoked histopathological alterations and small intestinal length reduction were not altered by the sst4 deletion.ConclusionThis is the first evidence for sst4 expression in myenteric plexus neurons and its protective roles against chemically‐induced gastrointestinal mucosal damage and inflammation. This suggests the safety of sst4 agonist drug candidates in the gastrointestinal tract, and even their potential use for gastroprotective indication.Support or Funding InformationSupported by: GINOP‐2.3.2‐15‐2016‐00050 – PEPSYS, GINOP‐2.3.2 STAY ALIVE, EFOP‐3.6.2‐16‐2017‐00008, ÚNKP‐19‐3‐III‐PTE‐211
Pilot Study: The role of chronic stress on brain microvessel density in male and female mice
Authors: Jent Wendy , Burrage Emily N. , Price Christian , Colbentz Tyler , Childers Ryan , Bryner Randy , Chantler Paul D. 
Published: April 2020

The cerebrovasculature, an intricate configuration of blood vessels, is responsible for the exchange of blood to and from the brain. Reduction of cerebrovascular vessels can reduce or prevent the flow of nutrients & oxygen that is necessary for proper brain function and increase risk of cerebrovascular diseases. Chronic stress is also a risk factor to various cerebrovascular diseases.In this pilot study, brain microvessel density in both female and male chronically stressed mice were analyzed. Beginning at the age of 18 weeks, mice were subjected to the unpredictable chronic mild stress (UCMS) paradigm for 5 days a week/seven hours a day for 8 weeks. Upon euthanization brain samples were flash frozen and sectioned at 8um using a cryostat. Immunofluorescence was utilized to observe endothelial cells via a CD31 antibody. Images were taken using a Nikon Eclipse 800 at a 40X objective and analyzed and quantified for density using ImageJ.Preliminary data (n=3/group) suggest that the male mice have a 26% higher microvessel density compared to females (p<0.05). When stressed, males exhibit a 17% lower microvessel density whereas stressed females did not display a loss in microvessel density (p<0.05).Here we showed that the brain of male and female mice can be physiologically diverse in microvessel density and that males and females display a different response when exposed to chronic stress.Support or Funding InformationNIH Cobra Grant 5P20GM109098; NIH INBRE P20GM103434
Probing the Role of Chloride in Facilitating System xc− Function
Authors: Catalfano Kevin , Chase Leah 
Published: April 2020

System xc− is a heterodimeric transporter that functions as a Na+‐independent antiporter exchanging extracellular cystine for intracellular glutamate. The transporter functions as an obligate heterodimer and is comprised of two proteins, xCT, which functions specifically in amino acid exchange, and 4F2HC, which appears to play a role in transporter stability. System xc− belongs to the SLC7 family of transporters, which is further subdivided into the cationic amino acid transporters (CAT) and the L‐type amino acid transporters (LAT), the latter of which is the subfamily which includes System xc−. In recent years, there have been advances in our understanding of the structure of the SLC7 transporters as crystal structures of bacterial homologs of CAT and LAT transporters have been published. As a result, we can now begin to ask important questions about the mechanism by which System xc− exchanges cystine and glutamate across the membrane. Previous studies in the Chase lab have demonstrated that System xc− is Cl−‐dependent. More recently we have shown that transport of one molecule of cystine requires more than one Cl− ion, and that Cl− binding affects the affinity of the transporter for cystine. We have also shown that chloride dependence of cystine transport is pH dependent. The K0.5 of chloride increases as the pH increases, suggesting that the Cl− binding affinity decreases at higher pH. Using this collective information, we examined the structure of xCT to identify the most likely binding site for cystine and glutamate. Specifically, we used Chimera 1.13.1 and Pyrx to dock cystine and glutamate into its most energetically favorable positions on xCT and identified several ionizable side chains within the putative permeation pathway associated with transmembrane domains 3 and 6 that appear to interact with substrate. These residues include R135, T138, Y246 and Y251. We are currently assessing this model in the presence of Cl− to identify putative residues which may be important for the docking of Cl− near the substrate binding site and using site‐directed mutagenesis to determine whether these residues are responsible for Cl−‐dependent cystine transport.Support or Funding InformationThis research is supported by the Campbell Foundation and Schaap Endowed Funds for Undergraduate Research.