bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differential white blood cell numbers in response to lipopolysaccharide exposure in marine and terrestrial mammals
Authors: Zenteno-Savín Tania , Symon Taryn Elizabeth , Reyes-Ramos Carlos Armando , Lugo-Lugo Orlando , Gaxiola-Robles Ramón , Vázquez-Medina José Pablo , Ramírez-Jirano Luis Javier , Hernández-Camacho Claudia Janetl 
Published: April 2020

Marine mammals, such as bottlenose dolphins (Tursiops truncatus) are often seen with wounds, bruises or shark bites down to the muscle, apparently with no signs of infection. In terrestrial mammals, including humans, similar injuries would be associated with infections, inflammation and potentially death. The objective of this study was to compare the differential leukocyte numbers in marine and terrestrial mammals under control conditions and following exposure to lipopolysaccharide (LPS). Blood samples were collected following routine procedures from healthy human anonymous volunteers from the blood bank at Instituto Mexicano del Seguro Social, La Paz, Baja California Sur, Mexico, and from healthy dolphins at CaboDolphin. All samples were taken with ethylenediaminetetraacetic acid (EDTA) as anticoagulant, transported to the laboratory on ice and processed within 24 h. Leukocytes were separated by centrifugation with ficoll hypaque and subjected to cell culture at 35°C with 5% CO2 and constant humidity. Leukocytes (106 cells) from both species were exposed to 10 μg LPS μL−1 for 24 or 48 h and number of each leukocyte cell type was registered; cell type was assigned based on morphological characteristics. Under control (no LPS) conditions, number of lymphocytes from both species was higher at 24 and 48 h as compared to the initial numbers. In humans, but not in dolphins, lymphocyte numbers were lower in cells exposed to LPS for 24 or 48 h as compared to untreated cells. In both species, at all times and regardless of exposure to LPS, an inverse relationship was observed between lymphocyte and neutrophil numbers. In humans, basophil number was lower in cells exposed to LPS for 24 and 48 h as compared to untreated cells. In T. truncatus only one cell with morphology similar to basophils was observed in only one individual. These differences in leukocyte types between species in response to LPS exposure could contribute to explain the apparent lack of infection or inflammation in injured marine mammals. This research was conducted in accordance with institutional ethical standards and following the 1964 Declaration of Helsinki and later amendments and in accordance with the FASEB Statement of Principles for the use of Animals in Research and Education. The research protocol (2018‐785‐010) was approved by the Ethics Committee of Instituto Mexicano del Seguro Social (CONBIOÉTICA‐09‐CEI‐009‐2016061).Support or Funding InformationConsejo Nacional de Ciencia y Tecnología (CONACYT CB‐2016‐01‐283669)
Discovery of First‐in‐Class Small Molecule Adhesion GPCR Modulators
Authors: Vizurraga Alexander , Yeung Jennifer , Tall Gregory 
Published: April 2020

GPR56 (ADGRG1) is a member of a subset of G protein coupled receptors called adhesion GPCRs (AGPCRs). These GPCRs are unique in their possession of an autoproteolytic extracellular domain that cleaves the receptor prior to activation. GPR56 is a model AGPCR that couples to G12/13 family G‐proteins and we recently discovered that it functions as a platelet collagen receptor important for hemostasis. Based on experimental evidence thus far, we believe that GPR56 is a first‐acting collagen sensor for platelets – enabling them to first recognize blood vessel injury and to initiate the platelet program that leads to repair. Despite being biologically relevant in a number of cellular processes and diseases, there is a distinct lack of available small molecule tool compounds for AGPCRs, including GPR56. We conducted a large‐scale high throughput screen to find agonists and antagonists of GPR56. Over 160,000 compounds were screened in conjunction with UM’s Center for Chemical Genomics (CCG) using a cell‐based serum response element (SRE) luciferase gene reporter assay in HEK293T cells. Primary screening yielded ~30 promising candidate full agonists and 8 candidate antagonists that robustly activated or inhibited GPR56 in cell‐based and biochemical assays. These compounds are currently being used in orthogonal platelet activation assays. Knowledge of these receptors is still sparse; the discovery of high affinity agonists and antagonists will be useful probes to explore AGPCR activities in endogenous contexts. These compounds may also serve as leads for the development of GPR56‐targeted pro‐ or anticoagulants.Support or Funding InformationGM120110, NS103946, T32‐GM007315
Assessing the ABCG2 Protein Interactome: a Potential Strategy to Disrupt ABCG2 Function
Authors: McHarg Ailsa Gavan , Gose Tomoka , Schuetz John D. 
Published: April 2020

The ATP‐binding cassette sub‐family G member 2, ABCG2, is an important plasma membrane transporter that removes potentially harmful endogenous toxins and xenobiotics from cells. ABCG2 was identified as an important contributor to the multidrug resistance phenotype in cancer cells because it exported a wide variety of structurally diverse chemotherapeutic agents, including mitoxantrone, topotecan and the tyrosine kinase inhibitors. High ABCG2 expression in a variety of tumours is associated with poor clinical outcomes. In our present study, we hypothesized that proteins that interact with ABCG2 modulate its function. This knowledge would allow us to discover novel therapeutic targets that could be exploited to alter ABCG2 expression and/or function.To identify interacting proteins for ABCG2, we employed a site‐specific biotinylation system utilizing a 15 amino acid peptide tag known as AviTag, fused to ABCG2. In the presence of the E. coli biotin ligase (BirA), biotin can be covalently attached to the AviTag peptide and the complex can be purified using the non‐covalent streptavidin‐biotin interaction. A plasmid harboring both AviTag‐ABCG2 and an IRES directed BirA was constructed. We stably expressed it in HEK293 cells and confirmed that AviTag ABCG2 was expressed at the plasma membrane and retained typical ABCG2 transport activity. Importantly, the cells expressing the AviTag‐ABCG2 and BirA readily labelled AviTag‐ABCG2 with biotin. The AviTag‐ABCG2 protein complexes captured by streptavidin agarose beads were purified and eluted according to an empirically developed protocol.Using a proteomic approach with mass spectrometry, we identified 39 proteins (both known and novel ABCG2 interacting proteins) at a p‐value threshold of 0.05 in the AviTag‐ABCG2 protein complex compared to the negative control. As expected, ABCG2 was one of the top 10 proteins in abundance (high spectral counts). Among the 39 interacting proteins, we confirmed by immunoblotting that several important cancer‐related proteins interact with ABCG2. These proteins appear capable of regulating ABCG2 function in cancer cells.Future studies will elucidate how these interactions affect ABCG2 function. These findings might aid in the development of new strategies for blocking ABCG2 function in multidrug resistant cancers, which could have important implications in improving patient’s response to chemotherapy.Support or Funding InformationThis work was supported by NIH and ALSAC
Evaluation of the mitragynine content, levels of toxic metals and the presence of microbes in kratom samples purchased in the Western Suburbs of Chicago
Authors: Prozialeck Walter C. , Edwards Joshua R. , Plotkin Balbina J. , Sigar Ira , Grundmann Oliver , Veltri Charles A. 
Published: April 2020

Kratom (Mitragyna speciosa) is an herbal product, indigenous to Southeast Asia, that has both stimulant and opioid‐like effects. Kratom leaf products are being increasingly used in the West for their reputed benefits in the treatment of pain and opioid use disorder. Recently, the US FDA and CDC have raised concerns regarding the contamination of some kratom products with toxic metals (Pb and Ni) and microbes such as Salmonella. To further explore this issue, 8 different kratom products were legally purchased from various “head”/”smoke” shops in the Western Suburbs of Chicago and then tested for microbial burden, a panel of metals (Ni, Pb, Cr, As, Hg, Cd), and levels of the main psychoactive alkaloid mitragynine. All of the samples contained significant, but variable, levels of mitragynine (3.9–62.1 mg/g) indicating that the products were, in fact, kratom. All but 2 of the samples tested positive for the presence of various microbes including bacteria and fungi. However, none of the samples tested positive for Salmonella. The same 6 samples that contained microbes also showed significant levels of Ni (1.7–7.9 μg/g), Pb (0.32–1.7 μg/g) and Cr (0.23–4.1 μg/g) while the other 2 samples were negative for metals. These data indicate that many kratom products contain variable levels of mitragynine, and can contain significant levels of toxic metals and microbes. These findings highlight the need for more stringent standards for the production and sale of kratom products.Support or Funding InformationThis work was supported by personal funds from Dr. Walter Prozialeck and by restricted funds from Midwestern University.
East Coast Versus West coast: Medical Student’s Anatomy Lab Experience
Authors: Nair Rakesh Ravikumaran , Mohan Gisha , Nausheen Fauzia 
Published: April 2020

ObjectivesAnatomy labs are traditionally taken as first operation room of medical students where future doctors make their first incision. Although, some new medical school are not dissection based but they implement many proactive pedagogies to make anatomy learning an interesting, interactive, engaging and exciting experience. We looked into similar teaching methodologies of two US medical schools : Florida International university‐COM and California University of Science and Medicine‐SOM.The objective of this study is to compare the student’s perception of anatomy lab teaching strategies of two medical schools based on student’s end of the course evaluations and final gradeMethodswe developed a collaboration between FIU‐COM and CUSM‐SOM anatomy faculty to share the curriculum, evaluation comments and performance of medical students after completion of Anatomy course. We are in the processes of getting exempt from IRB. The study data will include students feedback comments on teaching strategies and their final grade average for the course.ResultsAs a result of our collaboration, we reviewed the Course curriculum, evaluation comments and performance of medical students after completion of anatomy course between 2 universities. FIU curriculum is discipline based in year 1 and system based in year 2. The lectures are mostly didactic, and labs are led by the instructors. The teaching methodologies in lab include: a checklist of objectives, the identifications on plastic models, plastinated specimens and prosected cadavers. Students learn the structures and confirm their understanding of the anatomy with facilitators. During the discussion, facilitator review the high yield structures to the whole group using the DOC‐cam projector.CUSM‐SOM has a system‐based curriculum that is built on the active learning, student centered teaching strategies. Classes are mostly teams‐based and flipped classrooms. The labs are mostly student‐centered with multiple stations that student rotate every 20–30 min. One lab of 4 hours typically have 6–8 stations. Various aspects of anatomy of the same parts are learned at each station. These include: the gross anatomy specimens, embedded quizzes on specimens, videos, radiology images, hands‐on ultrasound experience and application questions. Final data analysis of the variables will be done after obtaining IRB exempt.ConclusionThe student evaluations from two different school’s anatomy lab teaching methodologies will hopefully identify the most effective teaching strategies that is most received and accepted by the students of this era. Collaborative efforts can improve anatomy lab teaching and student learning.
Regulator of G protein Signaling 10 (RGS10) functionally interacts with store‐operated calcium entry machinery to regulate COX‐2 and TNF‐α in Microglia
Authors: Wendimu Menbere Yisfashewa , Alqinyah Mohammed , Vella Stephen , Moreno Silvia N.J. , Hooks Shelley 
Published: April 2020

Activation of microglia is a driving factor of neuroinflammation and a hallmark of neurodegenerative diseases. The central inflammatory enzyme cyclooxygenase 2 (COX‐2) and the pro‐inflammatory cytokine tumor necrosis factor‐alpha (TNF‐α) mediate critical features of microglial activation responses, and their upregulation in the brain is a characteristic feature of many neurodegenerative diseases and brain injury. The Regulator of G protein Signaling 10 (RGS10), a small RGS protein enriched in microglia, has G protein‐independent functions in suppressing lipopolysaccharide (LPS)‐induced expression of COX‐2 and TNF‐α. Herein, we identify a biochemical and functional interaction between RGS10 and the endoplasmic reticulum (ER)‐localized calcium sensor STIM2, which is an essential component of the store‐operated calcium entry (SOCE) machinery. We determined that STIM2 and its downstream calcium pathway, particularly Orai channels and the Ca2+‐dependent phosphatase calcineurin, are essential for LPS‐induced COX‐2 and TNF‐α production in microglia, and likewise this pathway mediates the inhibitory effect of RGS10 on LPS‐signaling. We further demonstrated that ER calcium depletion amplifies LPS‐induced COX‐2 and TNF‐α expression and that RGS10 functions to suppress SOCE triggered by ER depletion. Collectively, this physical and functional interaction between RGS10 and STIM2 in microglia suggest a complex regulatory network connecting RGS10, LPS‐signaling, and SOCE, with broad implications in the pathogenesis and treatment of chronic neuroinflammation.Support or Funding InformationNIH‐NINDS
Gut to Eye Axis: Probiotic Mitigates Dysbiosis and Improves Ocular Functions in Mice
Authors: Singh Mahavir , Homme Rubens P. , Sandhu Harpal , George Akash K. , Stanisic Dragana , Malonee Carissa J. , Molnar Jack , Tyagi Suresh C. 
Published: April 2020

It is known that a dysbiotic gut together with bacterial infection can disrupt ocular functions. As demonstrated by our laboratory, and others that obese phenotype in mice is metabolically prone to ocular hypertension phenotype, including retino‐vascular occlusion. These physiological perturbations arising presumably from alterations in the microbial population(s) of the host lead to loss of ocular homeostasis. We thus hypothesized that restoration of the gut‐microbiome via long‐term probiotic administration may help improve ocular functioning that has been derailed by the dysregulated “1‐carbon metabolism” as a result of high fat diet (HFD) consumption. To test this hypothesis, we used 4 different experimental groups of mice, designated as: Group‐I: WT mice (C57BL/6J); Group‐II: WT + HFD, Group‐III: WT + HFD + PB (Lactobacillus rhamnosus), and Group‐IV: WT + PB (Lactobacillus rhamnosus). The PB (Lactobacillus rhamnosus) intervention was done orally employing 2.5x105 CFU/day for a continuous period of 16 weeks. Visual function analysis, and detailed retinal angiography of the WT + HFD mice revealed somewhat abnormal vascular patterns, and impaired ocular functions. These conditions were subsequently improved by PB (Lactobacillus rhamnosus) supplementation. Further experimentations using electro‐retinography (ERG), ocular coherence tomography (OCT), and the fluorescein angiography (FA) analyses on WT+ HFD + PB (Lactobacillus rhamnosus) treated mice and other experimental groups supported the observed beneficial effects. Taken together, our findings indicate that PB (Lactobacillus rhamnosus) treatment could restore ocular metabolism, in part, via mitigating the adverse retino‐vascular remodeling phenotype as induced by the dysbiotic environment, most likely, via modulation of the “gut‐to‐eye axis” through reconstitution of the gut‐microbiota.Support or Funding InformationHL139047 and AR071789
SERO‐PREVALENCE OF HIVANTIBODIES AMONG STUDENTS OF IMO STATE POLYTECHNIC UMUAGWO
Authors: Uchegbulam Henry Ugochukwu , Emeka-Nwabunnia (Nee Kalu) Ijeoma , Etteh Casmir Chinedu , Azuonwu Obioma , Nwankpa Promise , Ezekwe Ahamefula Sunday 
Published: April 2020

Human Immunodeficiency Virus (HIV) is the name given to the virus that causes HIV infection and it’s continuous attack on the body defensive mechanisms leads to Acquired Immune Deficiency Syndrome (AIDS). Since its discovery in 1981, the virus has been widespread and has shown to be of global public health concern, thus, empirical evidence strongly suggest that HIV attacks the CD4+T‐lymphocyte‐ a special type of T‐lymphocyte that fights off disease in our immune system and render them ineffective to perform it’s task as requiredThe aim of the study was to investigate the HIV Sero‐prevalence among students of Imo State Polytechnic Umuagwo extraction.A total of 1301 apparent healthy students aged between 19–24 years, were recruited through convenient random sampling technique, between June to November, 2009 respectively. However, out of these number, 536(41.20%) were males and 765(58.8%) were females and a rapid screening kit (Unigold kit‐Trinity Biotech, Ireland) which is based on antibody and antigen reaction was explored according to manufacturer’s instruction for the assay. The Microsoft excel software was used to represent the results in bar and pie chart respectively.The result from the assay showed that 11 (0.85%) of the males tested positive whereas 10(0.77%) were discordant while 18(1.38%) females tested positive whereas 8(0.62%) were discordant. These probably suggest that among the youths of these ages, females were more infected than the males. Thus, there is need for continues health education among youths as abstinence from premature sex and avoidance of risky habits could be a strong veritable tool of prevention among active youths in our global communities.
Cross Interaction Between OGT and mTOR in T Cells and its Consequences in Autoimmunity
Authors: Castrillo Savannah Danielle , Gorelik Gabriela 
Published: April 2020

BackgroundSystemic Lupus Erythematosus (SLE) is a chronic autoimmune disease that has been studied extensively, yet an understanding of the mechanisms of this disease remains unclear. It is known that epigenetic mechanisms are involved in the disease. We found O‐GlcNAc Transferase (OGT), an X‐linked gene, overexpressed due to hypomethylation in T cells from lupus patients (J. Autoimmunity, 2013). Furthermore, it was reported that the mechanistic target of rapamycin (mTOR) modifies T cell cytokine patterns and contributes to the TReg defect in patients with lupus. (J. Immunology, 2014).PurposeTo investigate the role OGT has in T cell signaling in order to understand the contribution of its overexpression to lupus. In particular, we investigated the relationship between OGT and the mTOR pathway in T cells to understand their participation in the disease.MethodsHealthy female donors were recruited by advertising. The research was reviewed and approved by the University of South Alabama Review Board for Human Subject Research. Peripheral blood mononuclear cells (PBMC) were purified by density gradient centrifugation, then T cells were isolated by magnetic beads. One fraction was transiently transfected for 48 h with 2μg of OGT siRNA using a nucleofector device (Amaxa). Other fractions were subjected to different treatments. Cells were stimulated after treatments with PMA/Ionomycin for 4h and the whole cell extracts were separated by SDS‐PAGE electrophoresis. Protein expression was analyzed by immunoblotting using specific antibodies. ß actin was used as loading control.ResultsIncreasing the levels of glycosylated proteins with 100 mM Pugnac, a competitive inhibitor of O‐GlcNAc‐β‐N‐acetylglucosaminidase (OGA), or with 5mM Glucosamine, increased the phosphorylation of p70S6K, substrate of mTORC1, which indicates an increase in mTOR activity. On the other side, when OGT is knocked down by siRNA, as expected, the protein expression of OGT levels as well as glycosylated protein levels decreased. Interestingly, the activity of mTOR also decreased. Similar results were observed with 10 nM MHY1485, potent activator of mTOR, or with 50 nM rapamycin and 10 nM Torin‐1, both inhibitors of mTOR, which increased or decreased the phosphorylation of its substrate respectively. However, they were unable to modify the levels of glycosylation.ConclusionsThese results strongly suggest that mTOR activation is OGT‐dependent and is located downstream of OGT in T cells. Overexpression of OGT could be responsible for the increase in mTOR activity in T cells from lupus patients.Support or Funding InformationNIH 5R03 AR067518‐02Summer undergraduate research fellowship (SURF) University of South AlabamaDepartment of Biomedical Science, College of Allied Health, University of South Alabama
Loss‐ And Gain of Function Mutations in the Type 1 IP3 Receptor (IP3R1) Modify Agonist‐Evoked Vasoactive Responses in Cerebral and Skeletal Muscle Resistance Arteries From Adult Mice
Authors: Mishra Ramesh C. , Zhu Hai-Lei , Chen S.R. Wayne , Cole William C. , Braun Andrew P. 
Published: April 2020

Calcium mobilization within vascular smooth muscle and endothelial cells of small resistance arteries plays a critical role in agonist‐evoked vasoconstriction and dilation, respectively. Despite the recognized contribution of IP3‐mediated intracellular Ca2+ release for agonist‐evoked vasoactive responses, it has been difficult to study this process acutely in native arteries, due to the lack of selective pharmacological agents or experimental strategies to target the predominant IP3R1. In humans, heterozygous mutations in the IP3R1 are known to produce either loss‐of‐function (LOF) or gain‐of‐function phenotypes for Ca2+ release that impact cerebellar activity (e.g. ataxia), yet it remains unclear how these global mutations affect the cardiovascular system. To facilitate such investigation, we have engineered novel genetic knock‐in mice harbouring a single amino acid mutation at position Asp2594 in the transmembrane S6 helix of the endogenous IP3R1 Ca2+ release channel that confers either a LOF (Asp2594Ala) or GOF (Asp2594Lys) phenotype, respectively. In permeabilized HEK293 cells expressing these mutant receptors, the LOF mutation shifted IP3‐mediated ER Ca2+ release ~2‐fold to the right, whereas the GOF mutation produced a 2.3‐fold leftward shift compared with wild‐type (WT) IP3R1. In endothelium‐denuded cerebral arteries pressurized to 10 mmHg, the sensitivity of serotonin‐evoked vasoconstriction was reduced in vessels from heterozygous LOF mice compared with WT, whereas agonist responsiveness was enhanced in arteries from heterozygous GOF mice. Vasoconstriction in response to the thromboxane receptor agonist U46619 was modestly enhanced by the GOF mutation, but largely unaffected in arteries from LOF mice. In cannulated cremaster skeletal muscle arteries from LOF and GOF mice pressurized to 70 mmHg, the sensitivity of endothelium‐dependent, acetylcholine‐mediated inhibition of myogenic tone was right shifted (3.7‐fold) and left shifted (2‐fold), respectively, compared with vessels from WT. Comparable changes in functional sensitivity were observed in response to the endothelium‐dependent vasodilator bradykinin. In contrast, dilation evoked by the NO donor sodium nitroprusside was not different among the three genotypes. Collectively, these data demonstrate that genetic modification of IP3R1‐mediated Ca2+ mobilization in small resistance arteries functionally impacts agonist‐evoked vasoactive responses. These observations highlight the importance of IP3R1 activity in the vascular wall, and further implicate vascular impairment as a contributing factor to the neurological dysfunction observed in patients with LOF and GOF IP3R1 mutations.Support or Funding InformationThis work was funded by the following research grants from the Canadian Institutes of Health Research: MOP‐97988 (WCC), MOP‐142467 (APB) and PJT‐155963 (WC, WCC and APB).