bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of COUP‐TFII in Glucose Homeostasis in Dahl Salt Sensitive Rats
Authors: Ashraf Usman Mohammad , Kumarasamy Sivarajan 
Published: June 2021

The Interaction between genetic and environmental factors plays an important role in disease progression. One of the major environmental factor includes diets, which governs significant attention based on the broad spectrum of effect in cardiovascular and associated diseases. Consumptions of diets that are rich in calories and salts can significantly increases the risk for developing life threatening cardiovascular events including hypertension and metabolic syndrome. Through GWAS studies, a locus with significant association for hypertension was found in human chromosome 15 containing the gene Chicken Ovalbumin Upstream Promotor Transcription Factor II (COUP‐TFII). To validate this gene, a mutant rat was developed with a 15‐base pair deletion in Dahl Salt Sensitive (SS) rat background. Under high salt diet regimen, the Coup‐TfIImutant rat had an improved cardiac and renal function when compared to SS rats. Being a known transcriptional regulator, Coup‐TfII also plays a role in other physiological events including glucose homeostasis and energy metabolism. In H4IIE rat hepatoma cells, Coup‐TfII expression modulated in response to glucose levels; under high glucose condition Coup‐TfII expression was significantly down‐regulated (p< 0.05); whereas in low glucose condition Coup‐TfII expression was significantly up‐regulated (p< 0.05). In addition, Coup‐TfII protein expression is found to negatively correlate with insulin regardless on the amount of glucose present in H4IIE cells (p< 0.05). Similar to our in‐vitro findings, Coup‐TfII gene expression was significantly up‐regulated in SS rat liver during fasting as compared to re‐fed condition (p< 0.05). Coup‐TfIImutant rat have also showed a significant decrease in fasting blood glucose levels (p< 0.05) compared to SS rat along with a significant decrease in their body weight (p< 0.05). Furthermore, the Coup‐TfIImutant rat demonstrated a superior glucose response and insulin response as measured by glucose tolerance test (GTT) and Insulin tolerance test (ITT) (p< 0.05). Further, under fasting condition the mRNA expression analysis in livers of SS and CoupTfIImutant rats revealed that the key enzymes involved in both beta oxidation and gluconeogenesis was significantly down regulated in Coup‐TfIImutant rats compared to SS rats (p< 0.05). Overall, our preliminary findings suggest that Coup‐TfII is important for glucose homeostasis in SS rats. Further understanding of Coup‐TfII function will help to dissect the relationship between metabolic syndrome and blood pressure regulation in SS rats.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TFAM Overexpression Prevents Skeletal Muscle Atrophy
Authors: Theilen Nicholas T , Jeremic Nevena , Weber Greg , Tyagi Suresh 
Published: June 2021

The current study aims to investigate if the overexpression of the mitochondrial transcription factor A (TFAM) gene in a transgenic mouse model diminishes soleus and gastrocnemius atrophy occurring with hindlimb suspension (HLS). Additionally, we aim to observe if combining concurrent, treadmill exercise training with TFAM transgenic mice prior to HLS has a synergistic effect in preventing skeletal muscle atrophy. Male transgenic mice aged 12–14 weeks old overexpressing the TFAM gene were assigned to a control (T‐Control), 7‐day HLS (T‐HLS), and 14 exercise sessions prior to 7‐day HLS (T‐Ex+HLS) groups and compared to male C57BL/6 (WT) mice (all wild‐type data from prior work currently in submission) aged 12–14 weeks old assigned to Control, 7‐day HLS (HLS), 14 exercise sessions prior to 7‐day HLS (Ex+HLS), and 14 exercise sessions (Ex). Results indicate overexpressing TFAM results in a decrease of 8.3% in soleus weight/bodyweight ratio and 2.6% in gastrocnemius weight/bodyweight ratio after HLS compared to losses in wild‐type mice of 27.1% in soleus weight/bodyweight ratio and 21.5% in gastrocnemius weight/bodyweight ratio after HLS. Our data indicates TFAM may play a critical role in protecting skeletal muscle from disuse atrophy and is correlated with increased expression of antioxidants (SOD‐2) and potential redox balance. Our results also indicate combining exercise with TFAM overexpression had no significant increase compared to wild‐type Ex+HLS. TFAM may be a molecule of interest for future pharmaceutical and therapeutic targeting to treat skeletal muscle atrophy.Support or Funding InformationThe study was supported by NIH Grant HL74185 to SCT.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PfaR Purification Strategy and Structure Analysis
Authors: Ortiz‐Rosario Melissa Cristina , Rullán‐Lind Carlos , Pérez‐Oquendo Mabel , Morales‐Lozada Yermary , González‐Méndez Ricardo , Baerga‐Ortiz Abel 
Published: June 2021

The biosynthesis of polyunstaurated fatty acids (PUFA) in certain bacterial species, is regulated by processes that are not fully understood. One possible factor in the modulation of gene expression in PUFA biosynthesis is pfaR, a putative transcriptional regulator whose gene is encoded directly upstream of pfaA in some bacterial species but entirely absent in others. Here we report the expression of pfaR in Escherichia coli together with its purification by Ni2+ affinity chromatography. Our results show that the majority of pfaR is produced as an insoluble protein that can be reconstituted from inclusion bodies. After inclusion body reconstitution, pfaR remains amenable for affinity purification. Additional bioinformatics analyses indicate the presence of a conserved winged helix‐turn‐helix (wHTH) domain flanked by a number of conserved motifs which include a predicted protein‐protein interaction site. Sequence alignments and motif structure analyses reveal evolutionary divergence in this family of transcriptional regulators, consistent with the likely DNA‐binding function of pfaR. Currently, we are in the process of analyzing pfaR secondary structure using circular dichroism. Future work will include functional analysis using systematic evolution of ligands by exponential enrichment.Support or Funding InformationThis work was supported by the National Institutes of Health [R25 GM061838, R25GM061151‐ 15, 5T34GM007821‐37, T36‐GM‐095335]; and the NationalScience Foundation [CHE0953254].This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Induced pluripotent stem cells derived from different tissue sources and their prospect for osteochondral regeneration
Authors: Danisovic Lubos , Csobonyeiova Maria , Novakova Zuzana Varchulova , Nicodemou Andreas , Varga Ivan 
Published: June 2021

A novel approach for stem cell generation is the attempt to induce conversion of the adult somatic cells into pluripotent stem cells so called induced pluripotent stem cells (iPSCs) by introducing specific transcription factors. iPSCs have two essential cell characteristics, they are pluripotent and posses long term cell‐renewal capacity. Additionally, iPSCs can be derived from patient‐specific somatic cells, thus bypassing ethical and immunological issues. The aim of present study was to reprogram human neonatal fibroblasts and adipose stromal cells by episomal reprogramming system. Obtained iPSCs were characterized by methods of molecular biology and microscopy. Distinct colonies of iPSCs started to appear between 14 – 20 days reprogramming and were proved by alkaline phosphatase (AP) live staining. After manual picking the colonies and their subsequent passaging, they did not lose ability to form embryoid bodies, they were positive for AP, Tra‐1‐60, and SSEA‐5. Moreover, obtained iPSCs expressed pluripotency markers Oct4, Sox2 and Nanog, and the expression levels of chondrogenic and osteogenic markers were significantly higher in comparison to control (normal fibroblasts). In summary, we have demonstrated neonatal fibroblasts and adipose tissue stromal cells can be reprogrammed into iPSCs. They shared similar biological and morphological features, and display strong potency of chondrogenic and osteogenic differentiation. Based on obtained results it can emphasized that after further analysis concerns on their biological safety they may be used as patient‐specific cells in regenerative medicine of osteochondral tissues.Support or Funding InformationSupported by grant APVV under contract No. 14‐0032 and by grant VEGA under contract No. 1/0153/15.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐inflammatory Peptide Ac‐SDKP is Highly Concentrated in Macrophages of Lymph Nodes
Authors: Romero Cesar Andres , Kumar Nitin , Carretero Oscar Alfredo 
Published: June 2021

N‐acetyl‐seryl‐aspartyl‐lysyl‐proline (Ac‐SDKP) is a natural peptide released from its precursor Thymosin β4 (Tβ4) by meprin‐α and the prolyl oligopeptidase enzymes (POP) in secuential step. Ac‐SDKP is hydrolyzed by the angiotensin converting enzyme (ACE). Ac‐SDKP has anti‐inflammatory and anti‐fibrotic effects in heart, aorta and kidney and part of the beneficial effects of ACE inhibitors has been ascribed to the increased levels of Ac‐SDKP. Ac‐SDKP is present in spleen and thymus, however, the presence of Ac‐SDKP in the lymph node has not been studied. We hypothesized that Tβ4, Ac‐SDKP and its releasing enzymes are present in lymph node. Ac‐SDKP concentration was evaluated in Sprague‐Dawley (SD) rat tissues. Tβ4 as well as Meprin‐α and POP enzymes were measured in the lymph node, thymus, spleen and other tissues obtained from SD rats. Ac‐SDKP was measure in several lymph node cell populations obtained by cell sorting. Lymph node showed the highest Ac‐SDKP concentration (181±18.3 ng/mg of protein) followed by the testis, thymus and spleen in SD rats (P<0.001 lymph node vs. testis, thymus and spleen). Meprin‐α and POP activity were present in lymph node, spleen and thymus. Tβ4 and Meprin‐α immunostaining were found to be positive in multinucleated giant cells in the cortical region of lymph node and along the septums; it was also found in the follicular region and germinal center. POP staining was found positive in the cortical region. Co‐localization of POP and the CD68 protein was found in the cortical region but not in the medulla. In the lymph node Ac‐SDKP concentration was higher in Macrophages (CD45+CD68+) in comparison with T lymphocytes (CD45+CD3+) (1.5±1.1*10−3 pmol/cell vs. 0.3±0.1*10−5 pmol/cell, respectively). We conclude that Ac‐SDKP is highly concentrated in lymph node and that all the components of Tβ4/Ac‐SDKP system are present. The releasing enzyme POP is present in Macrophages and these cells have the higher Ac‐SDKP concentration. Our present work provide new information that would be useful in inflammatory regulation in lymph nodes.Support or Funding InformationNational Institutes of Health Grant P01HL028982This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Novel, Long‐Lasting μ Opioid Receptor Antagonist Prevents and Reverses the Respiratory‐Depressant Effects of Heroin in Rhesus Monkeys
Authors: Gerak Lisa R , Maguire David R , Woods James H , Husbands Stephen , France Charles P 
Published: June 2021

Opioid overdose continues to be a serious public health problem in the US with fatal overdose occurring in at least 115 Americans every day. Naloxone (Narcan®) is increasingly available to reverse respiratory depression, the fatal effect of opioids; however, its relatively short duration of action limits the protection it can provide. SH‐1 is a novel opioid antagonist that has a longer duration of action than other opioid antagonists, including naloxone and naltrexone. The current study examined the ability of SH‐1 to prevent and reverse the respiratory‐depressant effects of heroin using head plethysmography to monitor minute volume (VE) in 4 monkeys breathing air. First, SH‐1 was given before heroin to determine whether it would prevent respiratory depression. The duration of action of SH‐1 was assessed by generating heroin dose‐effect curves at various times (1, 2, 4, and 8 days) after administration of 0.32 mg/kg SH‐1; these effects of SH‐1 were compared to those of naltrexone (0.032 mg/kg). The ability of SH‐1 to reverse the respiratory‐depressant effects of heroin was determined by giving a single injection of heroin to decrease minute volume (VE) followed 30 min later by an injection of either saline or 0.32 mg/kg SH‐1. In the absence of drug (control), average VE (± SEM) was 1531±113 ml/min. Heroin dose dependently decreased VE, and the potency of heroin varied across monkeys with the dose needed to decrease VE to <70% of control ranging from 0.1–1 mg/kg. When given before heroin, SH‐1 and naltrexone prevented the respiratory‐depressant effects of heroin, shifting the heroin dose‐effect curve 6‐fold rightward in each monkey. SH‐1 continued to attenuate the respiratory‐depressant effects of heroin for at least 4 days after administration whereas naltrexone antagonized heroin only on the day it was administered. When heroin decreased VE to <70% of control, subsequent administration of SH‐1 reversed these respiratory‐depressant effects within 30 min. Thus, SH‐1 can both prevent and reverse the respiratory‐depressant effects of heroin and these effects last longer than those of naltrexone. Because of its sustained effects, the novel compound SH‐1 might provide greater protection against the fatal respiratory‐depressant effects of opioids, thereby improving treatment of opioid overdose.Support or Funding InformationSupported by NIH (R01DA005018) and the Welch Foundation (AQ‐0039).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
BMPR2 and TfR1 Interact to Each Other
Authors: Alanazi Madhawi S. , Mohildien Ashraf M. , Nauli Surya M. 
Published: June 2021

BMPR2 (bone morphogenetic protein receptor type‐2) and TfR1 (transferrin receptor protein‐1) play an important role in embryonic development and are involved in cancer formation. Our recent proteomic study indicates that these two proteins are highly expressed in renal epithelial cells, although their functions in the kidney is not well understood. We confirm that both proteins have interaction in our immunoprecipitation study. We also knockdown BMPR2 or TfR1 expression in renal cell epithelia. Compared to scrambled control, knockdown of either BMPR2 or TfR1 results in a cell migration defect. The knockdown cells also show abnormal proliferation and apoptotic indices. In summary, our studies indicate that BMPR2 and TfR1 may share a functional interaction with each other.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐tuberculosis Drug Discovery from Phenotypic High‐throughput Screening of Actinomycete Cultures
Authors: Grzelak Edyta Malgorzata , Gao Wei , Choules Mary P. , Wan Baojie , Wang Yuehong , Cheng Jinhua , Lee Hanki , Suh Joo‐Won , Pauli Guido F. , Franzblau Scott G. , Jaki Birgit U. , Cho Sang‐Hyun 
Published: June 2021

This report presents over a decade of efforts in anti‐tuberculosis (anti‐TB) drug discovery at the UIC Institute for Tuberculosis Research (ITR). During this period, a library of approximately 200,000 actinomycete‐derived extracts was screened directly against virulent Mycobacterium tuberculosis (M. tb) H37Rv using a luxABCDE reporter and the Microplate Alamar Blue Assay.1 The collection consisted of soil microorganisms from sites in Southeastern Asia with distinctive ecosystems and climate, e.g., polar, tropical, volcanic, and deserted areas. The extracts of interest were subjected to extensive biological profiling under replicating and non‐replicating M. tb conditions, against mono‐drug resistant M. tb strains, and panels of microorganisms other than mycobacteria. The aim was to identify and validate the extracts with the most interesting activity profile by fractionation and bioassay‐guided isolation of the active principles. New approaches were implemented not only for the biological activity testing but also for the isolation and structure identification of the potential leads.2 The established anti‐TB drug discovery platform led to the discovery of the two anti‐mycobacterial cyclic peptides, ecumicin and rufomycin, which both target ClpC1 in M. tb.Support or Funding InformationThe Next‐Generation BioGreen 21 Program (No. PJ01328403), Rural Development Administration, Republic of Korea; and T32AT007533 from NCCIH/NIH.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neuroinflammatory Responses in a Mouse Model of Tetramethylenedisulfotetramine‐Induced Status Epilepticus
Authors: Calsbeek Jonas , Gonzalez Eduardo , Boosalis Casey , Zolkowska Dorota , Rogawski Michael , Lein Pamela 
Published: June 2021

Tetramethylenedisulfotetramine (TETS) is a GABAA receptor antagonist that is considered a chemical threat agent due to its ease of synthesis and its potent bioactivity in humans. Acute exposure to TETS can trigger seizures that evolve into status epilepticus (SE), and death. The current medical countermeasure for treating acute intoxication with seizurogenic chemicals like TETS includes the administration of benzodiazepines such as diazepam or midazolam. When administered 40 min post‐exposure in a mouse model of TETS‐induced SE, these positive allosteric modulators of the GABAA receptor are effective in stopping seizures and reducing mortality from ~90% to ~25%. The goal of this project was to determine the efficacy of this intervention in mitigating the neuroinflammatory response following TETS‐induced SE, with the objective of determining a feasible window of time for therapeutic intervention. In the current study, adult male NIH Swiss mice were pretreated with riluzole (10 mg/kg, i.p.) 10 min prior to receiving a lethal dose of TETS (0.2 mg/kg, i.p.). Control animals received vehicle in place of TETS and benzodiazepine. Animals were continuously monitored by EEG and/or scored for seizure behavior to confirm that TETS exposure elicited electrographic and/or behavioral seizures. At 10 or 40 min after the first myoclonic body twitch, animals were administered either diazepam (1.8 mg/kg, i.p.) or midazolam (1.8 mg/kg, i.m.) to terminate the persistent seizures. At 72h post‐intoxication, animals were perfused with fixative and brains were removed for FluoroJade C (FJC) labeling to assess neurodegeneration, and for GFAP and IBA1 immunohistochemistry to quantify neuroinflammation. Relative to vehicle control, GFAP immunoreactivity was significantly increased in the piriform cortex of all TETS‐exposed groups regardless of time of benzodiazepine administration. Midazolam was not superior to diazepam in protecting against neurodegeneration or neuroinflammation. These findings do not support the hypothesis that earlier treatment with either benzodiazepine is more effective in mitigating TETS‐induced neuropathology.Support or Funding InformationSupported by the NINDS CounterACT Program (grant # U54 NS079202).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antiproliferative Effects of Metformin in Triple Negative MDA‐MB 231 Breast Cancer Cells Exposed to Glucose‐Starved And 2‐Deoxyglucose
Authors: Triggle Christopher , Majeed Yasser , Samuel Samson Mathews , Ding Hong Mathews 
Published: June 2021

IntroductionWe have previously reported that 50μM of the anti‐diabetic drug metformin protects endothelial cells against hyperglycemia‐induced endothelial dysfunction and senescence thus providing support for the vascular protective benefits of metformin when used in diabetes 1,2. However, when 2mM (but not 50μM) metformin was used in a cell culture protocol with tumor murine microvascular endothelial in a glucose‐starved (GS) or 2‐deoxyglucose (2DG; 5mM) media pro‐survival autophagy and the mTOR pathway were inhibited and cell viability was reduced 3. These data supported other published evidence from both clinical and ex vivo cell‐based studies suggesting the utility of metformin for the treatment of various cancers. We therefore postulated that triple‐negative breast cancer (TNBC) that does not benefit from hormonal or HER2 targeted therapies may also respond to a metformin and GS/2DG protocol and investigated the effects of metformin in MDA‐MB 231 TNBC cells that were cultured in the presence of 2DG.MethodsMDA‐MB 231 cells were treated with 2DG (10mM) in a glucose free media for 48h in the absence/presence of metformin (2mM). Western blot analysis (n=3–4); cell proliferation (MTT assay; n=5); migration (Radius Cell Migration Assay; n=4) and viability (propidium iodide assay; n=5) were performed. Significance was determined by ANOVA.ResultsCell proliferation rate significantly decreased (95%) in 1) GS + metformin when compared to non‐treated GS (88%) and 2) 2DG exposed cells treated with metformin (46%) versus cells that were treated with either metformin (20%) or 2DG (28%) alone. Cell viability significantly (P<0.05) decreased in GS treated with metformin (34%) when compared to non‐treated GS (8%). In addition cell migration in cells exposed to 2DG and treated with metformin was reduced. Western blot analysis revealed that treatment with metformin significantly decreased (fold change) the levels of LC3A‐II (2.4&1.8) and LC3B‐II (2.3&1.6), pmTOR (S2448; 1.6&1.4) and downstream p4E‐BP1(T36/47; 1.5&1.3), pS6(S235/236; 4.7&2.8) and pS6(S240/244; 3.2&1.8) in GS and 2DG‐exposed cells + 2mM metformin.ConclusionResults show that using mM metformin in TNBC cells in a GS media containing 2DG offers a potential adjunct anti‐cancer protocol for the treatment of TNBC.Support or Funding InformationSupported by a Qatar Foundation National Priorities Research Program grant (NPRP: 04‐910‐3‐244) and a Junior Scientist Research Experience Program grant (JSREP: 03‐016‐3‐009)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.