bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dynamic L‐type CaV1.2 channel trafficking facilitates CaV1.2 clustering and cooperative gating
Authors: Ghosh Debapriya , Nieves‐Cintrón Madeline , Tajada Sendoa , Brust‐Mascher Ingrid , Horne Mary C. , Santana Luis F. , Navedo Manuel F. 
Published: June 2021

The L‐type CaV1.2 channel plays an essential role in gene expression, cell excitability and muscle contraction. These channels clustered throughout the plasma membrane of excitable and non‐excitable cells. This has been suggested to contribute to regional variations in channel activity and the concerted opening and closing of adjacent CaV1.2 channels (e.g. cooperative gating), but whether dynamic intracellular and near‐membrane trafficking of the channel modulates these features is unknown. Here we show dynamic transport of CaV1.2 in vesicles of circular and tubular shape with diverse intracellular and submembrane trafficking patterns. Microtubules and actin filaments are required for dynamic movement of CaV1.2 vesicles, which undergo constitutive homotypic fusion and fission events to sustain CaV1.2 clustering, channel activity and cooperative gating. In conclusion, our study reveals very diverse and unique intracellular and near‐membrane dynamics of the CaV1.2 channel in which homotypic vesicle fusion may be a key event promoting CaV1.2 membrane organization and clustering to facilitate its physical proximity for cooperative gating and fine‐tune control of Ca2+ signals.Support or Funding InformationNIHRO1HL098200, NIHRO1HL121059This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The regulation of muscle growth by miR‐23a
Authors: Beach Cassandra Lynn , Petrocelli Jonathan , Stott Nicole , Marino Joseph 
Published: June 2021

BackgroundMicro‐RNAs (miRNAs) are a class of small, non‐protein coding RNAs that posttranscriptionally regulate gene expression through the degradation or translational inhibition of their target complimentary messenger RNAs (mRNAs). Developing evidence indicates the importance of miRNAs in the myogenic program and their influence on disorders affecting muscle growth and metabolism.PurposeThe purpose of this study was to determine whether miR23a regulates myotube formation in skeletal muscle under lipid‐induced atrophic conditions.MethodsC2C12 myoblasts were transfected with a mimic complex to increase levels of miR23a or vehicle at seeding, then induced to differentiate into myotubes. Cells were treated with palmitic acid (0.1mM) at day 0 and day 2 of differentiation. Cells were collected at day 4 of differentiation. Real‐time PCR was used to investigate changes in miRNA expression and myosin heavy chain staining was performed to determine differences in myotube formation.ResultsCells that received mimic had increases in miR‐23a levels. Palmitic acid‐induced reductions in myotube formation were abrogated with overexpression of miR‐23a.ConclusionsThese data suggest that miR23a may exist in a regulatory pathway that alters muscle growth in the presences of saturated fatty acid. Future work is ongoing to understand the mechanisms by which miR23a may regulate skeletal muscle growth under lipid‐induced atrophic conditions.Support or Funding InformationFaculty Research GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Palmitic Acid on MicroRNA‐26a Expression and Skeletal Muscle Insulin Signaling
Authors: Petrocelli Jonathan Joseph , Beach Cassandra , Stott Nicole , Marino Joseph 
Published: June 2021

BackgroundMicroRNAs (miRs) are non‐protein coding RNAs that interact with messenger RNAs (mRNAs) to post transcriptionally regulate protein translation. The miRs important in regulating skeletal muscle insulin resistance are not fully identified.PurposeThe purpose of this study was to determine the role of miR‐26a on lipid induced insulin resistance in skeletal muscle cells.MethodsC2C12 mouse skeletal muscle cells were treated to increase levels of miR‐26a (mimic) using commercially available kits (Qiagen). C2C12 mouse myoblasts were grown to full confluence, serum starved for 3 hours then treated with 0.5 mM palmitic acid (PA) to induce insulin resistance. Cells received PA for 24 hours followed by a 100 nM insulin treatment for 15 minutes. Real‐time PCR was used to examine levels of miR‐26a, and Western blotting was utilized to investigate insulin sensitive protein expression.ResultsCells receiving mimic had increases in miR‐26a levels and cells receiving palmitic acid had decreased responsiveness to insulin. Overexpressing miR‐26a levels in C2C12 muscle cells improved responsiveness to insulin in the presence of palmitic acid.ConclusionsHere we show that increasing miR‐26a levels in vitro can improve insulin responsiveness in mouse skeletal muscle cells with lipid induced insulin resistance. The mechanisms of how miR‐26a is operating in skeletal muscle need to be further explored.Support or Funding InformationFaculty Research GrantThis 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 17β‐Estradiol on Endothelial Cell Expression of Inflammation‐Related miRNAs
Authors: Levy Ma’ ayan V. , Hijmans Jamie G. , Stockelman Kelly A. , Brewster Lillian M. , Reiakvam Whitney R. , Goldthwaite Zoe A. , Greiner Jared J. , DeSouza Christopher A. 
Published: June 2021

MicroRNAs (miRs) are short single stranded noncoding RNAs that are involved in the regulation of a number of physiological and pathological processes. miRs down regulate target gene expression post‐transcriptionally by degrading messenger RNA and/or by blocking translation. It is now recognized that miRs play a key role in regulating inflammatory processes underlying cardiovascular disease (CVD). For example, altered expression of specific miRs, such as miR‐126, miR‐146a, miR‐150 and miR‐181b have been linked with heightened vascular inflammation, atherosclerosis and CVD risk. It is well established that 17β‐estradiol is anti‐inflammatory and vasculo‐protective. Indeed, 17β‐estradiol‐mediated reductions in vascular inflammation have been linked with reduced pathogenesis of CVD. However, the mechanisms by which 17β‐estradiol influences vascular inflammation are not fully understood. Accordingly, the aim of this study was to determine the effects of 17β‐estradiol on endothelial cell expression of miR‐126, miR‐146a, miR‐150 and miR‐181b. Cultured human umbilical vein endothelial cells (HUVECs) were plated at a density of 5.0×105 cells/condition. Cells were treated with media alone or media containing 17β‐estradiol (100 nM) for 48 h. To determine miR expression, RNA was extracted from 1.0×105 cells and quantified using RT‐PCR. Cellular expression was normalized to RNU6 and calculated as fold change in ΔΔCt from control (N=4, experimental units). Cellular expression of miR‐126 (2.40±0.56 fold), miR‐146a (2.40±0.58 fold), miR‐150 (2.01±0.36 fold) and miR‐181b (2.47±0.56 fold) was significantly increased (240%, 240%, 200% and 250% respectively) in response to 17β‐estradiol treatment compared with control. These data indicate that 17β‐estradiol beneficially affects cellular expression of inflammation‐associated miRs. Higher cellular expression of miR‐126, miR‐146a, miR‐150 and miR‐181b is associated with reduced vascular inflammation by targeting nuclear factor κB (NFκB) activity. The alteration of the cellular expression of these miRs may play a role in the anti‐inflammatory and vasculo‐protective effects of 17β‐estradiol.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of miR‐101‐3p Prevents Human Aortic Valve Interstitial Cells Calcification through Regulation of CDH11/Sox9 Expression
Authors: Chen Jianglei , Lin Yi , Sun Zhongjie 
Published: June 2021

BackgroundCalcific aortic valve disease (CAVD) is one of the most prevalent forms of cardiovascular disease in the elderly population. There is no effective treatment for CAVD due to unknown mechanism,. The purpose of this study is to investigate whether miR‐101‐3p plays a role in human aortic valve interstitial cell (HAVIC) calcification and elucidate the underlying mechanism.Methods and ResultsSmall RNA deep sequencing and qPCR showed a significant increase of miR‐101‐3p in HAVICs isolated from calcified human aortic valves compared with normal valves. In vitro study using cultured primary HAVICs showed that mimic of miR‐101‐3p promoted calcification and upregulated osteogenesis pathway, while inhibition of miR‐101‐3p in the osteogenic conditioned medium prevented calcification and inhibited the osteogenic differentiation in the HAVICs. We found that miR‐101‐3p directly targeted CDH11 and Sox9 expression which were also downregulated in the calcified human aortic valves and HAVICs. Inhibition of miR‐101‐3p restored the CDH11 and Sox9 expression in calcific condition, downregulated asporin expression and prevented osteogenesis in the HAVICs.ConclusionsUpregulation of miR‐101 plays a crucial role in the calcification process in the HAVICs by directly regulating the expression of CDH11 and Sox9. Inhibition of miR‐101 prevents the calcification in the HAVICs and downregulates asporin through restoring CDH11 and Sox9 expression. Thus, miR‐101 may be an effective therapeutic target for CAVD.Support or Funding InformationNIH R01 HL118558, AG049780, DK093403This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The human immune response to RSV includes a loop between the tumor suppressor p53 and expression of APOBEC3 genes
Authors: Gladwell Wesley , Menendez Daniel , Li Heather , Yost Oriana , Resnick Michael A. , Kleeberger Steve 
Published: June 2021

BackgroundRespiratory syncytial virus (RSV) infects nearly all children by theirsecond year of life with re‐infections occurring throughout adolescence andadulthood. RSV infection typicallyresults in mild host symptoms. However, susceptible individuals can develop severe lower respiratory tract infectionsand bronchiolitis, sometimes leading to hospitalization or death. Recently we established that in addition toits protective role against cancer development, the tumor suppressor p53 alsocontributes to host immune responses regulating the expression of several genesincluding the APOBEC3 family of DNA cytidine deaminases (A3), consideredsentinels in innate immune response against viral infections.ObjectiveUsinghuman lung cancer cells, we tested the role of p53 in response to RSV infectionand its replication through the regulation of the A3 gene family as well asother components of the immune response.MethodsA549 and H1299 cells were infected with RSV (Strain A2). Cells were thenharvested 72–120 hours post‐infection and qPCR and Nanostring arrays? were usedto evaluate mRNA expression of RSV‐g, P53, A3s and other immune targets. Expression of RSV was assessed? by ConfocalMicroscopy and cytofluorometry while Annexin V/PI assay was used to evaluateapoptosis.ResultsIn response to acute and chronic RSV infection in human lung cancer cells, p53 was activated and had an impact on RSV replication, which involves theexpression in a p53 dependent manner of several members of the A3 gene familyas well as other innate immune genes. APOBEC3G (A3G) was highly induced in a dose‐ and time‐dependent mannerafter RSV only in lung cancer cells harboring WT p53. Accordingly, overexpression of A3G in lungcancer cells reduced RSV replication. Furthermore, a Nanotstring transcriptome analysis (immune code set) after RSV infectionrevealed a strong p53 dependent regulation of immune response genes, includingseveral components of the innate immune antiviral response. In addition, RSV infection induced p53phosphorylation at serine 15 and triggered p53‐dependent apoptosis in p53 functional cells while p53 deficiency correlated with increased viralreplication and increased syncytia phenotype.ConclusionsOverall, our findings establish that the “guardian of the genome” role ascribed to p53 also extends to a unique component of the immune system – the A3 genes – revealing novel components in the antiviralimmune response against RSV infection.Support or Funding InformationSupported by NIEHS Division of Intramural Research 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.
Do Omega‐3 Fatty Acids Prevent the Development of Nonalcoholic Fatty Liver Disease?
Authors: Gagen Ashley N. , Schaller Megan L. , Maciejewski J. Claire , Sandoval Karin E. , Witt Ken A. , Wooten Joshua S. , Guilford Brianne L. 
Published: June 2021

The prevalence of obesity in the United States is at an all‐time high. It is estimated that 80–90% of the obese population has nonalcoholic fatty liver disease (NAFLD), which is characterized by excess lipid accumulation in the liver (when lipid exceeds >7% of total liver mass). NAFLD can progress to nonalcoholic steatohepatitis (NASH) that includes development of hepatic inflammation, fibrosis, and cell death. Omega‐3 polyunsaturated fatty acids found in fish oils exhibit beneficial effects on regulating inflammation and may hold therapeutic potential as a preventative treatment of NAFLD.PURPOSEAssess the preventative effects of omega‐3 fatty acids on the development of NAFLD in high‐fat fed mice.METHODSMale C57BL/6 mice were randomized into four diet groups (n = 12/group) for 32 weeks: 10% low fat fish oil (LFFO), 41% high fat‐fish oil (HFFO), 10% low fat‐lard (LFL), or 41% high fat‐lard (HFL). At study completion, mice were euthanized and livers were frozen in O.C.T. compound in 2‐methylbutane while submerged in liquid nitrogen and stored at −80°C. Livers were cryosectioned at a thickness of 10 μm at −21°C. Fluorescence immunohistochemistry using Nile Red then allowed for the microscopic localization and quantification of neutral lipid droplets. The area of positive staining was calculated as a percentage of total section area, and an average neutral lipid droplet size was determined using Nikon Elements. A glucose tolerance test was performed at end study and the trapezoidal method was used to calculate total area under the curve (AUC).RESULTSBody weight in the HFL group was significantly greater than all other groups but glucose total AUC was only greater in HFL compared LFFO (p < 0.05). In addition, body weight (p < 0.05) and fasting glucose (p < 0.05) was greater in HFFO compared to LFFO. Based on this data, we expect a diet rich in omega‐3 fatty acids will mitigate hepatic lipid accumulation during prolonged feeding of a high fat diet. It is expected that HFL will have significantly greater liver lipid accumulation than HFFO, LFFO, and LFL.CONCLUSIONSThese findings would suggest a protective effect of omega‐3 fatty acids on development of NAFLD.Support or Funding InformationSupported by NIH R21NS090282‐01This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects Of Dates (Phoenix Dactylifera L.) On Basal And Stimulated Gastric Acid Secretion Due To Histamine, Pentagastrin And Carbachol In Rats
Authors: Onwuchekwa Chinedu , Okon Bassey 
Published: June 2021

Local folk claims that dates (Phoenix dactylifera L.) has a gastric beneficial role in humans. This study was carried out to examine the effects of dates on basal and stimulated gastric acid secretion (GAS) in the Wistar rats. Each group of rats consisting of 6 rats (180 – 210 g) were treated with aqueous undialyzed extract of dates (1.5 mg/kg, 2.5 mg/kg and 4 mg/kg) orally for 21 days. At the last day of the treatment, the rats were fasted for 24 hours, anaesthetised with ketamine. GAS ( mEq/l) induced by histamine, pentagastrin and carbachol administration were assessed by the continuous perfusion technique. Data were analysed using student's t‐test and ANOVA at p= 0.05.The aqueous extract of undialyzed dates significantly decreased (p<0.05) GAS induced by the histamine, pentagastrin and carbachol in a dose‐ dependent manner over 80 minute's period in all the treated groups. The gastroprotective action of dates may be multifactorial and this may include an anti‐secretory action.Support or Funding InformationUsmanu Danfodiyo University, Sokoto NigeriaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Canonical and non‐canonical inflammasomes are differentially regulated in NAFLD, which may play important roles in sensing hyperlipidemic DAMPs and initiating the pathogenesis of the disease and liver inflammation
Authors: Drummer Charles Edward , Johnson Candice , Wang Hong , Yang Xiao‐Feng 
Published: June 2021

Nonalcoholic fatty liver disease (NAFLD) is becoming increasingly common in countries with a Western‐style high fat diet. The caspase‐1‐dependent inflammasome pathway contributes to the pathogenesis of NAFLD and other chronic liver diseases. Current understandings are that caspase‐1 and caspase‐4 work synergistically in the canonical inflammasome pathway. However, recent research indicates that caspase‐4 can be activated independently and activate the non‐canonical inflammasome pathway. While the pathogenesis of the canonical inflammasome pathway in NAFLD has been studied, the non‐canonical inflammasome pathway is not well understood in this severe liver disease. We hypothesized that the non‐canonical pathway has its own up‐stream and down‐stream mediators, which may be independent of that associated with the canonical pathway. To test our hypothesis, we performed experimental data mining in various NAFLD NIH‐NCBI‐GEO microarray datasets. Our analysis revealed the following findings: 1) there is no overlap of the top ten upregulated pathways between the canonical and non‐canonical inflammasome pathways; 2) there is no overlap of the top ten downregulated pathways between the canonical and non‐canonical Inflammasome pathways; 3) the top pathways specifically mediating upregulation of non‐canonical pathways included two cell biology pathways apoptosis signaling and death receptor signaling; 4) the non‐canonical inflammasome pathways were metabolically related to adipose macrophage signaling (ATM) and docosahexaenoid acid (DHA) signaling; and 5) the non‐canonical inflammasome pathway was associated with down‐regulation of three retinoid × receptor alpha signaling pathways (FXR/RXR activation, LXR/RXR activation VDR/RXR activation). Of note, these nuclear receptors are metabolism master regulators, which were recently classified into our newly proposed anti‐inflammatory homeostasis‐associated molecular pattern (HAMP) receptors model (PMID: 29065888). Our results have demonstrated that canonical and non‐canonical inflammasome pathways are differentially regulated in NAFLD and that three retinoid × receptor alpha signaling pathways may specifically inhibit the upregulation of non‐canonical inflammasome pathways but not the canonical pathways. These findings have provided novel insights on differential regulatory mechanisms of canonical and non‐canonical inflammasomes in the liver and their potential roles in sensing hyperlipidemic DAMPs and initiating the pathogenesis of NAFLD.Support or Funding InformationNIH‐NHLBIThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sub‐organ Fractionation of Hepatic Cells after Antisense Oligonucleotide Treatment in Mice
Authors: Murray Heather , Peralta Raechel , Damle Sagar , Lee Richard , Prakash Thazha , Seth Punit , Fu Wuxia , Murray Sue , Guo Shuling 
Published: June 2021

Nonalcoholic Steatohepatitis (NASH) causes chronic liver disease and leads to advanced fibrosis and cirrhosis. Current research is unclear which liver cell population is key for treatment of the disease. Antisense oligonucleotide (ASO) drugs are a likely therapy for NASH due to their preferential distribution to the liver. Moreover, conjugates can be attached to the ASO and enable targeted cell specific drug delivery to the liver. Conjugations to the ASO also facilitate enhanced potency in targeted cell types by utilizing different uptake mechanisms. Unconjugated ASOs preferentially accumulate in the non‐parenchymal (NP) cells of the liver including liver sinusoidal endothelial cells (LSEC), Kupffer cells (KC) and hepatic stellate cells (HSC) before reaching the hepatocytes. Conversely triantennary N‐acetyl galactosamine (GalNAc)‐conjugation of ASOs results in a shift of ASO to preferentially accumulate in the hepatocytes over the NP cells (Prakash et al. NAR, 42: 8796, 2014). A recent study was designed to investigate the ASO activity in hepatocytes, LSEC, KC, and HSC from mice fed a standard chow diet compared to mice fed a Western diet. The liver pathology of the mice fed a Western diet at the time of ASO treatment was indicative of a NASH diseased state (steatosis, steatohepatitis and fibrosis). Mice were injected subcutaneously, at the indicated doses, with unconjugated or GalNAc‐conjugated ASO for 6 weeks. Following treatment, mouse livers were perfused, the hepatocytes were isolated by Percoll gradient and the NP cells by Optiprep (Lodixanol) gradient. NP cells were further isolated into enriched populations by fluorescence‐activated cell sorting (FACS). Whole liver and isolated cell populations were evaluated for RNA reduction after treatment. Drug concentration in each cell type was also measured using an ELISA based assay. As expected, there was target mRNA reduction with unconjugated and conjugated ASOs in all hepatocyte cell fractions. There was also targeted mRNA reduction in all NP cell populations in both healthy and diseased mice. However, the HSC cell population contained the least amount of ASO resulting in the lowest level of targeted reduction. These results suggest that GalNAc‐conjugated ASO can decrease targeted RNA not only in hepatocyte cells, but also in LSEC, KC and HSC cells despite a lower dose and a sever diseased state. This finding illustrates the ability of GalNAc‐conjugated ASO to enhance drug potency yet still allow drug delivery to all liver cell types. Therefore, ASO treatment is an attractive therapy for liver diseases, such as NASH, that involve multiple cell types contributing to disease progression.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.