bluuebunny/crossref_metadata_2025_split huggingface dataset:
Serine Rich Splicing Factor 2 (SRSF2) Flips for RNA Binding
Authors: Arnholt Mark , Bohn M. , Cina M. , Downing A. , Flanagan M. , Hautala L. , Jaeke A. , Kennedy C. , Lane B. , O'Bryon A. , Pepin J. , Rohloff E. , Rrahmani F. , Schmitt S. , Spudich J. , Staus S. , Tadlock K. , Tomashek M. , Witt E. , McNally Mark 
Published: June 2021

Mistakes in alternative splicing of RNA cause diseases such as acute myeloid leukemia. Serine Rich Splicing Factor 2 (SRSF2) is a splicing factor that controls alternative splicing by promoting exon inclusion, so it is not surprising that mutations to SRFS2 are linked to cancers. SR proteins harbor an RNA recognition motif (RRM) at the N‐terminus that binds to mRNA. SRSF2 has the unusual ability to bind to both pyrimidine and purine rich RNA sequences by flipping two C or G nucleotides in the mRNA into anti or syn orientations. The RRM specifically recognizes only C2, C3, and G5. Arg61 forms a hydrogen bond to C3, Phe59 hydrogen bonds to C2, and Lys17 is involved in flipping C2 or G2 into syn or anti conformations. Tyr92 binds with C2 and forms a hydrogen bond with C3. The mutation Pro95His binds better to UCCAGU and has been linked to cancer. The Hartford Union High School SMART (Students Modeling A Research Topic) Team modeled SRSF2 using 3D printing technology to highlight structural characteristics involved in RNA binding. Additional research on SRSF2 mutations and sequence binding has the potential to find both causes of and treatments for diseases like cancer.Support or Funding InformationThe MSOE Center for BioMolecular Modeling would like to acknowledge and thank the National Institutes of Health Science Education Partnership Award (NIH‐SEPA 1R25OD010505‐01) and the National Institutes of Health Clinical and Translational Science Award (NIH‐CTSA UL1RR031973) for their support in funding the 2017‐2018 SMART Team Team program.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Co‐Transcriptional Folding of a Riboswitch Controls the Fate of the Transcriptional Machinery
Authors: Walter Nils G. 
Published: June 2021

Folding of nascent transcripts can be modulated by properties of the RNA polymerase (RNAP) that carries out the transcription process. For example, the site‐specific pausing of the RNAP and co‐transcriptional RNA‐protein interactions have been shown in several cases to be important for co‐transcriptional RNA folding. In bacteria, riboswitches are genetic elements frequently found in the 5′ untranslated regions of mRNAs, where they respond to cellular metabolites to regulate gene expression either at the level of transcription or translation. Since they change conformation “on‐the‐fly”, riboswitches are tractable models for studying the role of the transcription process in guiding the folding of nascent transcripts.The preQ1 riboswitch from Bacillus subtilis regulates gene expression by a transcriptional mechanism in which the binding of its ligand, the queuosine precursor preQ1, causes a conformational change that leads to the formation of a terminator hairpin. Previous studies have deciphered the structure and dynamics of the isolated riboswitch aptamer, but less is known about the impact of the transcriptional machinery on riboswitch folding and vice versa. We have identified a strong pause by RNAP during riboswitch transcription that is stabilized by a previously characterized consensus pause sequence and the ligand‐free conformation of the riboswitch. By using single molecule Förster resonance energy transfer (smFRET) analysis of the riboswitch in a paused elongation complex, we show that RNAP has a profound impact on RNA dynamics, stabilizing a tightly folded structure at this specific pause site. Molecular modeling and biochemical assays further support a specific functional interaction between the nascent transcript and the exit channel of RNAP through which ligand binding to the riboswitch unpauses the polymerase. We are currently using cryo‐electron microscopy of the paused elongation complex to further pinpoint the specific structural rearrangements in both riboswitch and RNAP that are induced first by pausing, then by ligand binding. Finally, we are evaluating the impact of transcription factors on pausing and riboswitch folding. Our study provides significant insights into the mechanisms by which small RNA structures can regulate the function of the macromolecular transcription machinery, and vice versa.Support or Funding InformationNIH R01 grants GM062357, GM118524 and GM122803This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential Response of mTOR and ERK Signaling in Human Skeletal Muscle Following Resistance Exercise and Acetaminophen Consumption
Authors: D'Lugos Andrew C , Patel Shivam H , Fry Christopher S , Carroll Chad C , Dickinson Jared M 
Published: June 2021

Previous data have shown that cyclooxygenase (COX) inhibiting drugs may interact with the cellular mechanisms governing the adaptive responses of skeletal muscle to exercise. However, findings from acute exercise trials of attenuated growth responses with COX inhibitors do not translate to chronic exercise training, where COX inhibitors may augment muscle adaptation. The purpose of this study was to determine the extent to which prior consumption of acetaminophen (APAP) alters skeletal muscle signaling of the mammalian target of rapamycin (mTORC1) and ERK pathways in response to RE. In a double‐blinded, counter‐balanced, crossover design, healthy young men (n=8; 25 ± 1 yr, BMI: 26 ± 2 kg·m−2) performed two trials of unilateral knee extension (8 sets, 10 reps, 70% 1RM). For 24h prior to each trial, subjects ingested either APAP (1000mg/6h) or placebo (PLA) (final dose consumed immediately post RE). Muscle biopsies (vastus lateralis) were collected at rest and at 1 and 3h postexercise during each trial. Western blot was utilized to assess mTORC1 and ERK signaling proteins and immunohistochemistry was utilized to assess localization of mTOR to the lysosome (LAMP2). Localization of mTOR to the lysosome (LAMP2) increased in myosin heavy chain (MHC) II fibers 3h postexercise only in PLA (P<0.05). At 1h p‐rpS6Ser240/244 was increased in both groups, but to a greater extent in PLA (P<0.05). At 3h p‐S6K1Thr389 was elevated only in PLA, whereas p‐S6K1Thr421/424 was elevated only in APAP (P<0.05). Similarly, p‐ERK1/2Thr202/Tyr204 was increased to a greater extent in APAP at 3h (P<0.05). Consumption of APAP prior to resistance exercise blunts the translocation of mTORC1 to the lysosome and the phosphorylation of S6K1 at Thr389. Conversely, prior consumption of APAP augments ERK signaling following resistance exercise, leading to the phosphorylation of S6K1 at Thr421/Ser424. These data support a model in which ERK signaling may provide a compensatory response to inhibition of mTORC1 signaling. These findings also further highlight the intricate interaction between COX‐inhibiting drugs and the cellular processes regulating skeletal muscle adaptation to exercise.Support or Funding InformationSupported by intramural funds from ASU.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
rHbI‐S2H as a Potential Novel Hydrogen Sulfide Donor
Authors: Oliveras Alfredo Reyes , Arias Hazel Borges , López‐Garriga Juan 
Published: June 2021

Hydrogen sulfide (H2S) is a well‐known poisonous gas whose toxic effects have been studied for many years. In recent studies, it was found that H2S is produced endogenously in humans through enzymatic pathways and has functioned as a vasodilator, anti‐inflammatory, an antioxidant and smooth muscle relaxant. As result, current studies of H2S are focused as a signaling molecule involved in various physiological processes like NO and CO. Also, H2S concentration in human physiology has been associated with diseases like hypertension, Alzheimer's, cancer, arthritis, diabetes, ulcerative colitis and cardiovascular diseases. Biochemical and physiological studies of H2S have been performed by using compounds that release or promote the production of H2S in biological samples for its therapeutic attribution known as H2S donors. Hemoglobin I (HbI) is a hemoprotein found in the clam Lucina pectinata that has the property of binding H2S in its heme group forming the HbI‐SH2 complex. For this reason, the viability of the HbI from L. pectinata was studied for the delivery of H2S in biological systems to obtain a reliable H‐2S donor. Kinetics studies were performed using a UVV is spectroscopy at physiological conditions with the following parameters; Krebs' buffer at pH 7.4 and 25°C and 37 °C for 24 hours. During the 24 hours, the Soret band showed a displacement from 426 nm (HbI‐SH2) to 407 nm (HbImet) indicating the release of H2S from the protein. The half‐life of the complex was approximately 6 hours. The dissociation constant (koff) 0.43×10−3 seg−1. This result shows the high affinity of the H2S for the protein regardless of physiological pH and buffer.Support or Funding InformationThis work was funded by the National Science Foundation STC award number 1231306. Research reported in this publication was supported by the National Institute Of General Medical Sciences of the National Institutes of Health under Award Number T34GM008419. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Potential Influence of Follistatin and Myostatin on Body Composition during the Yukon Arctic Ultra
Authors: Johannsen Michelle , Schalt Adriane , Kim Jimin , Chen Richard , Coker Melynda S. , Gunga Hanns‐Christian , Coker Robert H. , Steinach Mathias 
Published: June 2021

ObjectiveThe objective of this study was to determine the potential roles of follistatin and myostatin on body composition under conditions of prolonged physical activity, chronic cold exposure and remote isolation.MethodsParticipants traveling on foot in the 2017 Yukon Arctic Ultra 692‐km (430‐mile) were recruited for the study. Measurements and samples were obtained at pre‐event, mile 173 at Carmacks checkpoint 1 (C1), mile 239 at Pelly Crossing (C2) and post‐event. Body composition measurements were obtained using bioelectrical impedance analysis. Wrist worn accelerometer devices were utilized to provide an estimation of caloric expenditure and dietary recall provided an assessment of caloric intake. Blood serum samples were collected, centrifuged on‐site, frozen and later analyzed using enzyme linked immunosorbent assays to determine myostatin and follistatin concentrations. Results were analyzed using one‐way ANOVA, presented as means±SEM and considered significant at P<0.05.ResultsTen participants (37±10 yr, body mass index: 24.4±2.6 kg/m2; 8 males, 2 females) were recruited, and five male participants completed the entire event in 260±9 hours. Caloric intake/expenditure was 4,244±471 kcal/day and 6,669±427 kcal/day, respectively, indicating a caloric deficit of 2,425±653 kcal/day. Total mass, body mass index and fat mass were reduced at each time point of the event. Fat free mass was unchanged throughout the event (Pre‐event: 60±3 kg, C1: 60±3 kg, C2: 60±3 kg, Post‐event: 59±1 kg). Follistatin was increased at C1 (1,715±310 pg/mL) in comparison to all other time points. Myostatin trended towards an increased level at C1 (20,494±5,531 pg/mL).ConclusionsDespite a remarkable caloric deficit and extreme cold conditions, fat free mass was preserved in participants traveling on foot in the event. Transient alterations in follistatin and myostatin may occur during chronic exercise and may potentially influence the regulation of lean tissue retention under these conditions. Future studies will be directed at the role of nutrient strategies and sleep deprivation on cytokines and physiological resilience under these conditions.Support or Funding InformationResearch reported in this publication was primarily supported by the DLR grant 50WB1330. Additional support was also provided by the National Institute Of General Medical Sciences of the National Institutes of Health under Award Numbers UL1GM118991, TL4GM118992, or RL5GM118990. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Skeletal Muscle Carnosine Content on Apoptotic Signaling
Authors: Church David D , Hoffman Jay R , Townsend Jeremy R , Jajtner Adam R , Beyer Kyle S , Oliveira Leonardo , Fukuda David H , Stout Jeffrey R 
Published: June 2021

Carnosine (Carn) has a pluripotent role in skeletal muscle (SkM) physiology, enhancing intracellular buffering, acting as an antioxidant, antiglycating and ion‐chelating agent, and also acts as a diffusible Ca2+/H+ exchanger. Carn role as an antioxidant may reduce apoptosis, but has not been fully investigated in human SkM. Therefore, the purpose of this study was to investigate if SkM Carn influences apoptotic signaling in response resistance exercise. Fourteen recreational active men completed a lower‐body resistance exercise protocol consisting of the squat, leg press, and leg extension exercises. Blood samples were obtained before (PRE), immediately‐(IP), one‐(1H), five‐(5H), 24‐(24H) & 48‐(48H) hours post‐resistance exercise. SkM biopsies were obtained from the vastus lateralis at BL, 1H, 5H, & 48H post‐resistance exercise. SkM carnosine content was assessed via HPLC. Participants were placed into either a high Carn (HI: n = 7; Carn = 12.25 ± 2.40 mmol·kg−1WW) or a low Carn (LO: n =7; Carn = 5.60 ± 1.27 mmol·kg−1WW) group for analysis. Multiplex signaling assay kits were used to quantify phosphorylation status of apoptotic markers (JNK, FADD, p53, BAD, Bcl‐2). Circulating measures of muscle damage [myoglobin (Mb)] and oxidative stress [8‐hydroxy‐2′ – deoxyguanosine (8OHdG)] were assessed via ELISA. To control for BL differences a repeated measures analysis of covariance was used to analyze SkM data. Area under the curve (AUC) analysis was determined by the trapezoidal method. Although no significant interactions between the groups were observed for FADD, BAD, p53, Mb, or 8‐OHdG (p > 0.05)in response to resistance exercise, significant interactions were observed for JNK (p = 0.019) and Bcl‐2 (p = 0.032). Post‐hoc analysis indicated a significantly greater response at 1H for HI compared to LO in JNK (p = 0.035), while a trend towards an increase (p = 0.065) was observed for Bcl‐2. In addition, changes in Bcl‐2 and JNK following exercise were only significant in HI (p = 0.009 and p = 0.007, respectively), but not LO (p = 0.370 and p = 0.369, respectively). AUC analysis for 8OHdG indicated that circulating concentrations for HI (6,107.5 ± 2,144.6 ng·mL−1·min−1) was significantly (p = 0.037, D = 1.25) less than LO (8,838.7 ± 2,222.3 ng·mL−1·min−1). In conclusion, our results indicated that participants with higher Carn have a reduction in pro‐apoptotic proteins, with an attenuated circulating oxidative stress response to exercise.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of endothelin type B receptors (EDNRB) on smooth muscle cells is controlled by ternary complex factors and the actin cytoskeleton
Authors: Krawczyk Katarzyna Karina , Skovsted Gry Freja , Perisic Ljubica , Rippe Catarina , Dreier Rasmus , Berg Jais Oliver , Lykkesfeldt Jens , Hedin Ulf , Swärd Karl 
Published: June 2021

Endothelin signaling plays an important role in physiology and disease and one of the endothelin receptors, the type B receptor (ETB or EDNRB), is known to be highly plastic, being upregulated in smooth muscle cells by arterial injury and following organ culture in vitro. Herein, we hypothesized that this transcriptional plasticity may arise in part because EDNRB is controlled by ternary complex factors and by the myocardin family coactivator MKL2. In line with this hypothesis we found significant correlations between the ternary complex factors ELK3, FLI1 and EDNRB, and between MKL2 and EDNRB at the mRNA level in human arteries. Overexpression of MKL2 in human coronary artery smooth muscle cells (HCASMC) led to a 30‐ fold induction of EDNRB, and this effect was antagonized by myocardin (MYOCD) which also correlated negatively with EDNRB at the tissue level. Overexpression of ELK3 and FLI1 induced expression of EDNRB, whereas the expression of classical smooth muscle markers was reduced. Depolymerization of actin using latrunculin B (LatB) increased EDNRB on both mRNA and protein levels, and increased calcium responses on stimulation with the ETB‐selective agonist sarafotoxin 6c (S6c) and endothelin 1 (ET‐1). Pretreatment with LatB in organ culture increased S6c and ET‐1‐induced contraction. We also found that ROCK inhibition using Y27632 induced EDNRB expression, and that the effect of LatB is antagonized by MAP‐kinase inhibition. Transcript‐specific primers and promoter reporter assays showed that the EDNRB_2 isoform is regulated by LatB in HCASMC. We finally demonstrate that the mRNA level of EDNRB is increased in rat carotid lesions and human carotid plaques, and that ELK3/FLI1 induction closely parallels EDNRB induction. In all, these studies uncover transcriptional control mechanisms for the endothelin type B receptor (EDNRB) that may explain its plasticity and could be targeted for therapy.Support or Funding InformationThis study was supported by grants from the Medical Faculty and The Royal Physiographic Society. KKK was supported by Marie Curie ITN Small Artery Remodelling of European Union.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Magnesium Deficiency Attenuates Vascular Smooth Muscle Contraction in Rat Aorta
Authors: IESAKI Takafumi , KAKIGI Ryo , WATANABE Makino 
Published: June 2021

Magnesium (Mg) deficiency has been reported to be associated with the development of various cardiovascular diseases including hypertension, however, vascular responses in Mg deficient state have not been fully elucidated. We have reported that Mg deficiency attenuates both voltage‐gated and receptor‐operated vascular smooth muscle (VSM) contraction without affecting maximal force development of contractile apparatus. The purpose of the present study is to further examine the mechanism(s) involved. Male Sprague‐Dawley rats were fed an Mg‐deficient diet (Mg content; 4.5 mg/100 g diet) and deionized water for eight weeks, and then the thoracic aorta were excised to measure isometric tension in organ chambers. CaCl2‐induced contraction in the presence of calcium (Ca2+) ionophore ionomycin under Ca2+‐free condition was significantly reduced in Mg deficient group (Mg‐Def). Prostagrandin H2 analogue U46619‐ induced contraction under Ca2+‐free condition was also greatly decreased in Mg‐Def, whereas direct protein kinase C activator phorbol 12,13‐dibutyrate‐induced contraction was not attenuated. These results suggest that Mg deficiency might impair, at least in part, the pathways which increase Ca2+ sensitivity of VSM.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelial dysfunction can be reversed in obese humans and mice by overexpression of endothelial Kir2.1 channels
Authors: Fancher Ibra S. , Adamos Crystal S. , Ahn Sang Joon , Phillips Shane A. , Levitan Irena 
Published: June 2021

The vascular endothelium is an important regulator of vascular tone. Acute changes in mechanical stressors, such as blood flow (fluid shear) and pressure, are converted into chemical signals by the endothelium to control vasomotor function. We recently showed that endothelial Kir2.1 channels are critical to the vasodilatory response to increases in blood flow through eNOS and NO production. It is well established that obesity causes endothelial dysfunction likely through impairment of nitric oxide signaling/bioavailability. Furthermore, obesity impairs the vasculature of visceral adipose depots while arteries in the subcutaneous adipose appear unaffected. Therefore, we tested the role of endothelial Kir2.1 in obesity‐induced endothelial dysfunction in vascular beds of spatially distinct adipose depots in obese humans and in a diet‐induced obese mouse model. Mice were fed a high fat, high cholesterol diet for 4 or 8 weeks prior to analysis of vascular function via response to flow in an ex vivo arterial preparation. Endothelial function of arteries from both subcutaneous and visceral adipose depots were tested. After 4 weeks of high fat feeding, no differences were observed in vascular function between arteries from either adipose depot nor when compared to control mice fed a normal chow diet. However, after 8 weeks visceral arteries from high fat fed mice had a significantly blunted response to flow, while subcutaneous arteries retained a full dilatory response. Overexpressing endothelial Kir2.1 using adenoviral transduction with expression driven by the VE‐Cadherin promoter in visceral arteries of diet‐induced obese mice rescued the response to flow suggesting that Kir2.1 channel function or expression may be impaired exclusively in arteries of visceral adipose of obese mice. Importantly, we observed similar findings in obese human arteries from visceral and subcutaneous adipose biopsies collected during bariatric surgery.Support or Funding InformationNIH R01 HL073965 (IL)American Heart Association 16POST27000011 (ISF)NIH 5T32HL007829‐24 (ISF)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.