bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bioinformatic Identification of Candidate miRNA Biomarkers for Traumatic Brain Injury
Authors: Smothers Christine , O'Connell Grant C 
Published: June 2021

The identification of peripheral blood biomarkers which are associated with traumatic brain injury (TBI) could lead to the development of molecular diagnostics which could be used to aid acute detection in scenarios where neuroimaging in unavailable, or to remotely track injury progression and recovery. It is becoming increasingly evident that brain tissue exhibits patterns of micro‐RNA (miRNA) expression which are distinct from those of other tissues. During TBI, the blood brain barrier becomes disrupted and molecules from damaged neural tissues are released into peripheral circulation. Thus, the detection of brain specific miRNAs in peripheral blood could serve as a surrogate marker of TBI. In this investigation, we aimed to systematically identify brain‐enriched miRNAs, and then test their potential utility for use as TBI biomarkers. First, we obtained publically‐available expression data for over 1,300 miRNAs generated from 31 different human post‐mortem tissues. Tissue specificity index (Tau) was calculated for each miRNA, and they were subsequently ranked in terms of their degree of brain‐specific expression. The abundances of the top five ranked miRNAs were then measured in serum samples collected from 10 TBI patients and 10 healthy controls using qPCR, and evaluated for their ability to discriminate between groups using k‐nearest neighbors (k‐NN). The top five miRNAs identified in our tissue specificity analysis consisted of miR‐137, miR‐219a‐5p, miR‐128‐3p, miR‐124‐3p, and miR‐138‐5p, which exhibited from 31 to 74‐fold higher expression levels in brain relative to other tissues. Three out of the five candidate miRNAs exhibited significantly higher abundance in serum samples from TBI patients relative to control patients in qPCR analysis, and their coordinate expression levels were able to discriminate between groups with 90% sensitivity and 80% specificity. The five candidate miRNAs identified in our analysis have potential utility for use as TBI biomarkers, and such a possibility warrants further investigation.Support or Funding InformationWork was supported by Case Western Reserve University FPB School of Nursing start‐up funds issued to Grant C. O'Connell.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Toxicity and Immunogenicity of a Tardigrade Cytosolic Abundant Heat Soluble Protein
Authors: Esterly Harrison James , Crilly Candice , Piszkiewicz Samantha , Pielak Gary , Christian Brooke 
Published: June 2021

Tardigrades are microscopic animals recognized for their resilience to extreme stresses, including desiccation, freezing, boiling, low and high pressures, and high doses of radiation. Their ability to survive desiccation is attributed to tardigrade‐specific cytosolic abundant heat soluble (CAHS) proteins. Expression of these proteins in other cells increases the desiccation tolerance of those species, and in vitro, CAHS proteins inhibit desiccation‐induced inactivation of other proteins. We are seeking to test the ability of CAHS proteins to stabilize protein‐based therapeutics, which require refrigeration for transportation and storage. The ability of CAHS proteins to preserve the activity of these pharmaceuticals upon desiccation at room temperature would increase their availability and reduce their cost. To this end, we investigated the toxicity and immunogenicity of CAHS D when injected into mice. Recombinant CAHS D was expressed in E. coli and purified by cation‐exchange chromatography. Purity was verified by mass spectrometry prior to endotoxin removal. Purified CAHS D was injected at various concentrations, and mice were monitored for symptoms of toxicity. No changes in weight or behavior were observed during the 28‐day monitoring period. A large‐scale toxicity study, as well as additional studies to test immunogenicity, is underway.Support or Funding InformationUniversity of North Carolina System Institutional Planning GrantThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Squishing Cellular H2O
Authors: Bartolomei Barrett , Bogdon Phoebe , Frazer Mandy , Johnson Robert , Lopez Dayana , Mergens Steven , Nichols Andrew , Owens Jonathan , Rall Tanner , Rhodes Abigail , Shipley Ashley Willow , Simmons Emily , Wright Zachary , Galea‐Martinez Sandra Ann , Bresniker Catherine 
Published: June 2021

Water is an essential part of the function of all living organisms. If water does not function normally, neurodegenerative and autoimmune diseases can become everyday occurrences affecting the health of humans. A research study was conducted about the structure and relative significance of the water channel, aquaporin 4 (AQP4), and it's possible conformational changes. Though structural details concerning the molecular mechanism of human AQP4 regulation are still unknown, this study has assisted in gaining insight into its structure and its interactions. The protein AQP4 consists of six‐transmembrane domains and five connecting loops that form a channel, and similar to other aquaporin channels, the monomers of AQP4 assemble into tetramers. AQP4 is namely arranged in clusters, known as orthogonal array particles, which allow for better anchoring to the membrane. Similar to other aquaporins, the pathway through the channel is amphipathic; this disposition allows for the bidirectional conductance of water. The physiological aspect (which is a 4‐fold axis structure) insulates against all solutes and water; the cytoplasmic side of the channel stabilizes the backbone amides. The mid‐membrane section, containing Phe‐195, Leu‐191, and Leu‐75, create a hydrophobic block (when repeated 4‐times). The research also revealed that AQP4 is subject to posttranslational regulation by phosphorylation. Water conductance is affected in AQP4 when SER‐111 (associated with the B loop) and Ser‐180 (associated with the D loop) are in a state of phosphorylation. Phosphorylated Ser‐180 is believed to possibly block the water channel by creating a tether to the C‐terminal domain. When Ser‐111 goes through phosphorylation, it mimics the gating mechanism in plants. This study of AQP4 suggests that the brain, spinal cord, and circadian rhythm are affected by its expression, as AQP4 is concentrated in the ends of astrocytes, of the brain. As new research comes to fruition, it is clear that these conformational changes will have broad implications with a variety of cell communication through that of water transport. The science community is on the frontier of research which could lead to therapeutic strategies and treatment of not only neurodegenerative diseases but autoimmune diseases as well. The El Capitan High School MSOE Center for BioMolecular Modeling MAPS Team used 3D modeling and printing technology to examine structure‐function relationships of Aquaporin 4.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ras and RTK: PI3K Activation, PIP3 Formation, Signal Transduction, Cancer Creation
Authors: Chou Chris , Lubkeman Kelly , Martyr Richard , Adler Rose , All Caleb , Arroyo Jorge , Austin Colby , Daniel Graham , Fehlberg Ezra , Grassmick Emma , Greer Maya , Hak Iain , Hubbell Grayson , Olmstead Grant , Perez Annette , Roberts Lauren , Skaggs Erin , Tafoya Frankie Rose , Whitted Jackson , Falke Joseph J 
Published: June 2021

Ras mutations are implicated in over 30% of all cancers. Under normal conditions, Ras becomes active by binding to guanine triphosphate (GTP) to stimulate cell growth and division and other cell pathways. When Ras acquires specific mutations, it no longer binds the GTPase‐activating protein (GAP), which impairs the GAP GTPase activity, trapping Ras in its active state and leading to uncontrolled cell proliferation. The Ras family of G‐proteins play a role in recruitment of phosphatidylinositol‐3‐kinase (PI3K) to the membrane and stimulation of PIP3 lipid signaling. The Ras family consists of 3 major isoforms: H‐Ras, K‐Ras, and N‐Ras; all having similar functions in different pathways. The primary sequence of amino acids among the Ras isoforms is highly conserved with differences existing primarily between their C‐terminal tails. The Falke lab is studying the H‐Ras variant. H‐Ras, in its active GTP‐bound state, and receptor tyrosine kinase (RTK) synergistically recruit PI3K to the membrane and stimulate PI3K kinase activity. During recruitment to the lipid bilayer, PI3K binds its substrate lipid phosphatidylinositol‐4‐5‐bisphosphate (PIP2) and also binds adenosine triphosphate (ATP) and then phosphorylates PIP2, creating phosphatidylinositol (3,4,5)‐trisphosphate (PIP3). PIP3 then plays a central role in many cell growth and regulatory pathways by recruiting to the membrane many proteins that bind to its headgroup. Specific surface amino acid sidechains on Ras that bind to specific surface amino acid sidechains on the Ras‐binding domain (RBD) of PI3K during their interaction to assemble the fully active PI3K complex were modeled. Drugs that inhibit Ras or PI3K or their interaction may be effective in attacking cancer cell growth. Ultimately, understanding Ras and the mechanism of PI3K activation promotes understanding of a large subset of human cancers and facilitates the development of targeted treatments. The Longmont High School SMART (Students Modeling A Research Topic) Team modeled wild‐type H‐Ras and oncogenic mutants using 3D printing technology to investigate structure‐function relationships.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Integrated of Sigma Quanta Energy in Psycho‐Neuro‐Physio Dynamics Therapy for Symptoms & Cause of Osteoarthritis: A Case Study
Authors: Sainvil Frantz Lee , Einstein George P , Konyk Carla M , Tulp Orien L , Rahman Michael S 
Published: June 2021

Osteoarthritis is the most common joint disease, afflicting some 27 million people in the United States alone. Over the past several decades, progress in the measurement of rates and interactions of molecular and cellular processes has initiated our understanding of dynamics phenomena of cells, Living cells are inherently dynamic. The vast qualitative information in the treatment of osteoarthritis is by allopathic medicine that can be upgraded by the individual algorithm of Sigma Quanta Energy in Psycho‐Neuro‐Physio Dynamics TherapyMethods and ResearchGenerally, the phrase dynamics phenomena refers also to Psycho‐Neuro‐Physiotherapies. The research on human subjects prepared algorithm therapy and life style. Algorithm of psychotherapy‐ by daily 15 min meditation, 30 minutes of Acu‐yoga exercises and 15 minutes meditation after. The effect of psychological intervention was also on regression ofAlgorithm of neuro therapy that emphasized nutrients for brain to support the threshold of the magnitude of stress intensity of an individual that must be exceeded for a certain reaction in meditation and Acu‐yoga exercise, phenomenon, result, condition to be manifested. Algorithm Physio‐Dynamics was design for application to microbial infection that gave a systematic delivery of complement inhibitors.ResultsThe aim of this research was determining the effect of Integrated of Sigma Quanta Energy in Psycho‐Neuro‐Physio Dynamics Therapy for Symptoms & Cause of Osteoarthritis. The application of the Triple Algorithm Dynamic therapy has been demonstrated to diminish or eliminate the osteoarthritic pain. There is a need to take daily a holistic approach and consider the impact of combinations of stress reduction, meditation exercise and dietary patterns, with a likely greater overall benefit than each single component might have on its own.Support or Funding InformationSupported by Institutional Resources of USAT, UHH, and EMIThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Upregulated miR‐144‐3p Attenuates LPS‐induced Acute Lung Injury via Inhibition of ROCK1 Signaling Pathway
Authors: Siddiqui Mohammad Rizwan , Tauseef Mohammad , Shahid Mohd. , Macdonough Kelli , Shanley Thomas 
Published: June 2021

Dysfunctional endothelial cell (EC) barrier and increased lung vascular permeability is a cardinal feature of Acute Lung Injury (ALI) and sepsis that may results in a pathophysiological condition characterized by alveolar flooding and pulmonary edema and subsequent hypoxemia. In lung ECs, activation of Rho associated kinase‐1 (ROCK1) phosphorylates myosin light chain (MLC), stress fiber formation and hyper‐permeability during ALI. The role of MicroRNA‐144 (miR‐144) has been well investigated in many human diseases including cardiac ischaemia/reperfusion‐induced injury, lung cancer and lung viral infection, however, its role in pulmonary EC barrier regulation remains obscure. Here, we investigated miR‐144 mediated mechanism in the protection of endothelial barrier function in bacterial wall lipopolysaccharide‐(LPS) induced lung injury model. By using transwell endothelial permeability assay to examine in vitro permeability and immunofluorescence microscopy to determine barrier integrity, we showed that ectopic expression of miR‐144 effectively blocked human lung EC barrier disruption and hyper‐permeability in response to LPS. Furthermore, using a gain and loss of function strategy, overexpression of miR‐144 significantly decreased ROCK1 expression. Concomitantly miR‐144 inhibits ROCK1 mediated phosphorylation of MLC phosphatase and thus phosphorylation of MLC to counteract actin stress fibers formation in LPS activated human lung EC. Finally, in mice challenged with LPS, in vivo delivery of miR‐144 mimic via liposomes attenuated endotoxemia‐induced increased in lung wet‐to‐dry ratio, vascular permeability and inflammatory cytokines. In conclusion, our data suggest miR‐144 attenuates activation of the inflammatory ROCK1/MLC pathway in vascular EC is a promising therapeutic strategy to counter inflammatory lung injury.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Stability Studies of Rooperol and Analogues by In Vitro Metabolism with HPLC/MS Detection
Authors: Bohanon Amanda L , Kerwin Sean M 
Published: June 2021

African Potato, Hypoxis hemerocallidea, has a long history of use by indigenous people of South Africa to treat cancer and a variety of other diseases. Extracts from the corm contain the norligan bisglycoside hypoxoside, which is hydrolyzed in the body to yield the anticancer agent rooperol (1,5‐bis(3′,4′‐dihydroxyphenyl)pent‐1‐en‐4‐yne). Studies have shown that rooperol selectively inhibits the growth of several cancer cell lines. Phase I clinical trials of rooperol in lung cancer patients suggested promising activity with no dose‐limiting toxicity. However, rooperol is rapidly converted to biologically inactive sulfate and glucuronide metabolites. Thus, a relatively small amount of the drug reaches the tumor site. Bioisosteric analogues of rooperol have been synthesized with the aim of increasing metabolic stability while preserving the anticancer properties of the parent drug. The goal of this research is to investigate the metabolic stability of rooperol and analogues. As part of this study, we employ an in vitro metabolism assay with porcine liver microsomes. Microsomes are supplemented with the cofactor UDP‐glucuronic acid and the pore‐forming peptide alamethicin. The suitability of this assay to characterize the Phase II metabolism of phenolic compounds was established with the plant phytochemical 3‐hydroxytyrosol. The time‐dependent metabolism of this compound was determined by HPLC assay, and the formation of the glucuronide Phase II metabolite was confirmed by HPLC/MS. The time course for the disappearance of 3‐hydroxytyrosol followed first‐order kinetics with an apparent half‐life of 65.4 minutes. We have recently employed this assay to quantify the metabolic lability of rooperol. The disappearance of rooperol monitored by HPLC revealed an exceptionally short half‐life of 3.3 ± 0.59 minutes and an intrinsic clearance of 0.101 ± 0.02 mL/min/mg protein. The identities of glucuronide metabolites of rooperol were verified by HPLC/MS. The extreme metabolic lability of rooperol demonstrates the need to identify more metabolically stable analogues. We will discuss the integration of this in vitro metabolism assay in a work‐flow designed to identify rooperol analogues exhibiting increased metabolic stability while retaining the cytotoxic activity of rooperol.Support or Funding InformationRobert Owens Medical Research FoundationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chondroitin Sulfate Pharmacological Activity from Four Different Sources in Human Osteoarthritic Cartilage
Authors: Montell Eulàlia , Pelletier Jean‐Pierre , Martel‐Pelletier Johanne 
Published: June 2021

ObjectiveChondroitin sulfate (CS) is a glycosaminoglycan and a well‐known component of cartilage extracellular matrix, which has been shown to have clinical benefit in symptomatic osteoarthritis (OA) and data suggest to be a modulating agent of joint inflammation. CS is generally produced by means of extraction from different animal tissues (bovine, porcine and marine cartilage). Because of differences in CS in terms of source, purity, quality and the production/purification process, we compared the effects of CS from four different sources on the function of human OA chondrocytes.MethodsExperiments were carried out on human OA chondrocytes obtained from knee patients undergoing total joint arthroplasty (n=8). Four different CS products were tested: CS#1, which is a high purity CS (98–100%) of bovine origin (Sioux Pharm Inc., Sioux Center, IA, US) and CS#2, CS#3 and CS#4, which are of lower purity and from other animal sources, CS#2 and CS#4, porcine; CS#3, avian. Determination of protein production of metalloproteinase (MMP)‐1 and MMP‐3, interleukin (IL)‐6 and prostaglandin E2 (PGE2) was performed on the conditioned medium using specific ELISAs. All determinations were performed in duplicate.ResultsCS#2 and CS#3 upregulated MMP‐1 (p=0.002, p=0.004) and CS#2 and CS#4, MMP‐3 (p=0.008, p=0.037). IL‐6 was also increased by CS#2, CS#3 and CS#4 (p=0.008, p=0.007, p=0.014). For PGE2 production, as the values were too low and unreliable without an inducer, the experiment was done only in the presence of IL‐1b (100 pg/ml), which greatly increased the value (absence of IL‐1b, 0.98±0.08 pg/mg protein; presence of IL‐1b, 42.18±10.80 pg/mg protein; p=0.007). CS#1 was the only one to significantly reduce PGE2 levels (p=0.035). In the presence of IL‐1b, CS#1 was also the only compound to show a trend toward significance (p=0.062) in reducing IL‐6 production.ConclusionsData showed that the three CS from non‐bovine origins increased an inflammatory factor in human OA chondrocytes, as well as upregulated the production of two catabolic factors. In contrast, CS of higher purity and coming from bovine source showed anti‐inflammatory properties as it reduced PGE2 and IL‐6 levels, even in the presence of IL‐1b. Therefore, the source/nature of these products could contribute to the overall biological and pharmacological effects of these agents in the symptomatic treatment of OA, compromising the safety and efficacy of the final products.Support or Funding InformationSupported by BIOIBÉRICA, S.A.U., Barcelona, Spain.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In vitro determination of the CB1 efficacy of illicit synthetic cannabinoids
Authors: Sachdev Shivani , Vemuri Kiran , Banister Samuel D. , Longworth Mitchell , Kassiou Michael , Santiago Marina , Makriyannis Alexandros , Connor Mark 
Published: June 2021

BACKGROUND AND PURPOSEThe recreational use of synthetic cannabinoid receptor agonists (SCRA) new psychoactive substances is associated with significant morbidity and mortality compared with herbal cannabis. To explore the relationship between SCRA CB1 receptor activity and acute toxicity, we determined the CB1 efficacy of thirteen SCRAs using receptor depletion with the irreversible CB1 antagonist AM6544 followed by curve‐fitting with the Black and Leff operational model. We also investigated the effects of brodifacoum, an anticoagulant associated with a recent outbreak of SCRA poisoning, on the acute signalling of human CB1 receptors.EXPERIMENTAL APPROACHReceptor depletion in mouse AtT‐20 pituitary adenoma cells stably expressing human CB1 was achieved by pre‐treatment of cells with AM6544 (10 μM, 60 mins). The CB1‐mediated hyperpolarisation of AtT‐20 cells was measured using fluorescence‐based membrane potential dye. The efficacy (tau) and affinity (KA) parameters were obtained for each drug by fitting data to the operational model. We next determined the effect of brodifacoum (1 μM, 5 mins) on the hyperpolarization induced by subsequent application of CP55940 or Δ9‐THC.KEY RESULTSThe tau value of Δ9‐THC was 80‐fold less than the reference CB‐agonist CP55940, and 260‐fold less than the highest efficacy SCRA, 5F‐MDMB‐PICA. The operational efficacy of SCRAs ranged from 233 (5F‐MDMB‐PICA) to 28 (AB‐PINACA), with CP55940 in the middle of the efficacy rank order. Application of brodifacoum had no effect on the potency, or the maximal response produced by high efficacy cannabinoid CP55940 or low efficacy phytocannabinoid Δ9‐THC (P<0.05).CONCLUSION AND IMPLICATIONSWe determined the efficacy of a library of the most prevalent SCRAs identified in the new psychoactive substances market since 2008 and found that the SCRAs we tested had up to 300 times the efficacy of Δ9‐THC, which may contribute to the apparently greater toxicity of these drugs. Our data suggests that cannabinoid induced signaling was not affected by brodifacoum, indicating that combining SCRA with brodifacoum is not likely to enhance user experience through interactions with CB1.Support or Funding InformationThis work was supported by NHMRC Project Grant 1107088 awarded to M.K., and M.C. and National Institutes of Health grant P01DA009158 to A.M.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.