bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterizing the Expression of TRPV4 in the Choroid Plexus Epithelia as a Prospective Component in the Development of Hydrocephalus in the Gas8GT Juvenile Mouse Model
Authors: Hochstetler Alexandra Elizabeth , Whitehouse Logan , Antonellis Patrick , Berbari Nicolas F. , Blazer‐Yost Bonnie L. 
Published: June 2021

Maintenance of homeostatic cerebrospinal fluid (CSF) secretion and absorption is essential for basic neurologic function. The choroid plexus epithelia, which is thought to produce the majority of the CSF, are among the most secretory of all epithelia. However, the control of this secretory process is poorly described. In diseased states such as hydrocephalus, where this homeostasis is disrupted, patients often experience symptoms such as cognitive impairment, motor/stability issues, and incontinence, among others. Current treatment for hydrocephalus is limited to surgically invasive shunting procedures which often fail and need revising – particularly in pediatric cases. In order to study the mechanism of this disease, a knock out mouse line of the Growth Arrest Specific 8 (Gas8) allele was generated. Loss of function of the Gas8 allele induces ciliopathy similar to Primary Ciliary Dyskinesia (PCD) in humans, with one of the symptoms being severe perinatal hydrocephalus. The disease progression in the mutant (−/−) mice is from postnatal day 0 (P0) to postnatal day 12–16 (P12–16), which makes this an excellent model to study pediatric/juvenile hydrocephalus. It is believed that one of the mechanisms of pediatric hydrocephalus is the overproduction of CSF by the choroid plexus (CP) epithelial cells that line the ventricles of the brain. The protein of interest to this research is the non‐specific cation channel, Transient Receptor Potential Vanilloid 4 (TRPV4), which has been shown to be activated by multiple stimuli including osmotic and pressure changes as well as by prostanoid metabolites. Activation of TRPV4 results in Ca2+ influx through the channel resulting in changes in intracellular signaling including the secondary activation of Ca2+‐stimulated ion transporters. Consistent with other studies, TRPV4 is localized to the apical membrane of the CP epithelial tissue in the Wild Type Gas8 mice. Preliminary data on the juvenile Gas8 mice indicate that TRPV4 is overexpressed in CP epithelia, ependymal cells and in the sub‐ventricular zone in the mutants as their hydrocephalus progresses as compared to the wild type (+/+) and heterozygous (+/−) animals. This suggests that antagonistic compounds of the TRPV4 channel have the potential to reduce hydrocephalus in the Gas8 model. Successfully targeting the molecular mechanisms for hydrocephalus in rodent models can provide a promising base for preclinical studies aimed at developing pharmaceutical agents to treat this disease.Support or Funding InformationFunding: Grants from the Hydrocephalus Association and the Department of Defense Office of the Congressionally Directed Medical Research Programs (CDMRP).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Whole‐genome DNA hydroxymethylation among monozygotic twins discordant for cardiovascular death: the prospective National Heart, Lung, and Blood Institute (NHLBI) Twin Study
Authors: Dai Jun , Leung Ming , Krasnow Ruth , Reed Terry , El‐Rifai Wael 
Published: June 2021

ObjectivesEpigenetic modifications are related to cardiovascular disease. It is unknown whether DNA hydroxymethylation is associated with the risk for cardiovascular death that is independent of genes and common environment. This study aims to address this issue.Methods19 male monozygotic twin pairs middle‐aged in 1969–1973 and discordant for cardiovascular death through December 31, 2014 were included from the prospective National Heart, Lung, and Blood Institute Twin Study. Buffy coat leukocyte DNA samples were collected in 1986–87. Genome‐wide DNA hydroxymethylation levels were quantified using the hMe‐Seal‐seq. The log2 transformed reads were used for analysis of differentially hydroxymethylated regions (DhMRs) across the genome. Benjamini‐Hochberg false discovery rate (q value) of 0.05 was considered as the significance level. The bioinformatic tools Enrichr with Gene Ontology (GO) and the Cytoscape software package were used to analyze biological functions.ResultsThere were 73,311 statistically significant DhMRs after controlling for age and blood cell subtypes (q<0.05 for all DhMRs). These DhMRs were located on all 22 autosomal chromosomes, and both the X and Y chromosomes. Among them, 27% of DhMRs were hypo‐hydroxymethlyated, had a log2 fold change ranging from −15.1 to −0.18, and were within/overlapped with 3′‐UTR (1.2%), 5′‐UTR (0.1%), exon (2.1%), intergenic regions (35.8%), intron (55.6%), non‐coding regions (0.6%), promoter transcription start sites (TSS) (2.4%), transcription terminal site (TTS) (2.1%); while 73% DhMRs were hyper‐hydroxymethylated, had a log2 fold change ranging from 0.14 to 15.7, and were within/overlapped with 3′‐UTR (0.9%), 5′‐UTR (0.1%), exon (1.7%), intergenic regions (49.2%), intron (44.9%), non‐coding regions (0.6%), promoter transcription start sites (TSS) (1.4%), and transcription terminal site (TTS) (1.2%). Using either all DhMRs with q value <0.05, a multi‐dimensional scaling plot demonstrated the complete distinction of case twins from control twins along dimension 1 but not dimension 2. A total of 12 significant GO biological processes (p‐value < 0.01) were identified. Cytoscape clustered the GO terms in these processes into 4 groups: target of rapamycin (TOR) signaling, metabolic process, cholesterol transport, and intracellular transport.ConclusionsDifferentially hydroxymethylated regions are prospectively related to cardiovascular death independent of genes and common environment.Support or Funding InformationNHLBI 1R21HL127368‐01 and Des Moines University Faculty Research Fund to Dr. DaiThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression Profile of G Protein‐Coupled Receptor 37L1 in mouse
Authors: Ma Xiaobo , Armando Ines , Jose Pedro A. , Konkalmatt Prasad 
Published: June 2021

The orphan G protein‐coupled receptor GPR37L1, also known as ETBR‐LP2, has strong homology with the endothelin‐B receptor (ETBR). However, GPR37L1 does not interact with endothelins. GPR37L1 is implicated to participate in brain development and the regulation of arterial blood pressure. GPR37L1 has been shown to be expressed in several regions in the brain, however its expression profile and the function in peripheral organs is poorly understood. We studied the expression profile of GPR37L1 in the brain, heart, kidney, liver, lung, spleen, stomach, pancreas, skeletal muscle, testis, and ovaries by RT‐PCR and immunoblot analyses and further used immunohistochemical analyses on tissue sections to identify the location of GPR37L1 expression within the tissue. Immunoblot analyses revealed the predicted 50 kDa and 40 kDa isoforms in most organs, and a ~35 kDa isoform mainly in the stomach and to a lesser extent in the brain, and a diffuse band of 45 kDa isoform mainly in the testis. The expression of GPR37L1 was highest in the brain, heart, stomach, pancreas, and skeletal muscle while GPR37L1 protein expression was lower in the kidney, liver, lung, spleen, testis, and ovary. The brain and pancreas expressed predominantly a 50 kDa isoform, while heart and skeletal muscle expressed mainly a 40 kDa isoform of GPR37L1. The kidney, liver, lung and ovary expressed almost equivalent levels of 50 kDa and 40 kDa isoforms of GPR37L1. The spleen expressed mainly a 40 kDa isoform of GPR37L1. RT‐PCR analyses confirmed the relative levels of GPR37L1 expression in the organs analyzed. Immunohistochemical analyses revealed the expression of GPR37L1, in the granule cell layer of the cerebellum, apical membrane of proximal tubule cells in the kidney, and airway epithelial cells in the lung. Further, RT‐PCR of proximal tubule and collecting duct cells obtained by laser capture micro‐dissection of mouse kidney sections, confirmed that the GPR37L1 is expressed in renal the proximal tubule but not in the collecting duct. These data show that GPR37L1 is expressed in the specific cell types within the organs and may play a key role in the physiology of the respective organs.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Enterocyte Lipid Droplet and Mitochondrial Interaction in Zebrafish (Danio rerio) Larvae During High Fat Diet Absorption
Authors: Walters James William , Compton Gregory , Andrews Lacey 
Published: June 2021

Diets associated with the development of dyslipidemias are typically high in triglycerides and cholesterol. These lipids are readily absorbed by intestinal enterocytes and there processed and packaged for delivery to the whole organism. Much remains unknown about the energetic mechanisms regulating dietary lipid processing within intestinal enterocytes. The purpose of this study is to better elucidate the dynamic interaction of lipid droplets and mitochondria during lipid absorption in the zebrafish model. Previous work in our lab has shown that larvae fed a high fat diet undergo lipid droplet biogenesis. We hypothesized that an increase in the size of the lipid droplets will cause mitochondria biogenesis or fusion while increasing apposition with lipid droplets. To test this prediction, zebrafish 6 day post fertilization larvae were fed a high fat diet of chicken egg yolk for one hour, rinsed, and fixed for TEM to create a 10 hour time course. Larvae were then sectioned, and imaged via TEM for each hour post feeding. Numbers of mitochondria decreased (43±7, 0 hours vs. 30±7, 3 hours) early upon feeding while area the total number of mitochondria increased (21±14 μm vs. 28±9 μm). Apposition between lipid droplets and mitochondria increased (from 0 vs. 5±1) during the same time frame and decrease by hour 10. Our results indicate that mitochondrial/lipid droplet apposition events correlate with lipid droplet processing or maturation. Future studies will attempt to further validate the zebrafish enterocyte model by comparison of other vertebrates. This work was supported by WV‐INBRE Grant# 2P20GM1.343_14 P1500699 to the West Virginia IDeA Network for Biomedical Research Excellence, NIH Grant P20GM103434Support or Funding InformationThis work was supported by WV‐INBRE Grant# 2P20GM1.343_14 P1500699 to the West Virginia IDeA Network for Biomedical Research Excellence, NIH Grant P20GM103434This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The DASH Diet Produces a Natriuretic Effect and Interacts with the Renin‐Angiotensin‐Aldosterone System
Authors: Maris Stephen A. , Brown Stacey , Conlin Paul R. , Williams Jonathan S. 
Published: June 2021

The DASH diet is widely recommended to lower blood pressure, but its mechanisms of action are unclear. Lines of evidence suggest an interaction with the Renin‐Angiotensin‐Aldosterone system (RAAS), including ACE‐inhibitor or natriuretic effects. The DASH‐Diet in Isolated Systolic Hypertension (DASH‐DISH) trial is a randomized, controlled, cross‐over, study to test RAAS‐related mechanisms underlying the DASH diet effect. Following a one‐week run‐in diet on a Control diet (CON), subjects consumed a DASH and a CON diet for 4 weeks each in a randomized sequence. Calorie content was controlled to maintain weight and sodium was set at about 3 g daily. After each diet, subjects were admitted to a clinical research center where hormonal and hemodynamic assessments were obtained supine at baseline and in response to a RAAS inhibitor—captopril 25mg (CAP). This created four study groups; 1) DASH, 2) DASH+CAP, 3) CON, and 4) CON+CAP. Each group was also studied for responsiveness to a RAAS stimulator—a graded angiotensin II infusion (1ng and 3ng/kg × 45mins each). Primary outcomes were renal blood flow (RBF), mean arterial pressure (MAP), plasma renin activity (PRA), and aldosterone (ALDO). A repeated measure ANOVA was computed for primary outcomes with a Bonferroni adjustment and a statistical significance set at .0125. A total of 44 (19 female) hypertensive subjects (age 55.7±13.6 yrs, BMI 29.7±5.2 kg/cm2, BP 149/82±8/6 mmHg) completed the study protocol. DASH diet showed a trend towards lowered MAP vs CON (96.1±11.7 mmHg vs 98.8±14.1 mmHg, p = 0.07) and DASH+CAP significantly enhanced effects on MAP vs CON+CAP (83.2±10.7 mmHg vs 87.8±14.3 mmHg, respectively, p < 0.01). There were no differences in baseline RBF between the DASH and CON (p = 0.17). However, DASH+CAP increased RBF more than CON+CAP (486.46±149.38 cc/min vs 451.21±171.11 cc/min, p < 0.001). There were no baseline differences in PRA between the DASH and CON (p = 0.92) but DASH+CAP significantly increased PRA vs CON+CAP (1.52±1.78 cc/ml/min vs. 0.89±1.17 cc/ml/min; p < 0.001). DASH showed greater ALDO sensitivity to AngII infusion when compared to CON (17.40±7.74 ng/ml vs 13.84±6.21 ng/ml, p < 0.05) as did DASH+CAP vs CON+CAP (15.06±5.32 ng/ml vs 13.08±5.89 ng/ml, p < 0.05). These results suggest that the DASH diet produces a natriuretic effect in lowering blood pressure and stimulates the RAAS. The DASH diet might be particularly helpful in lowering BP in populations who benefit from a diuretic/natriuretic response and/or are concurrently prescribed RAAS antagonists.Support or Funding InformationNIH NHLBI: R01 HL77234, NIH NHLBI: K23 HL084236This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mutations in the zebrafish ortholog of HCFC1 reveal a critical function in neural precursor function
Authors: Reyes Joel Fernando , Castro Victoria Lynn , Reyes‐Nava Nayeli G. , Quintana Anita M. 
Published: June 2021

Mutations in the HCFC1 transcriptional co‐factor cause cblX syndrome, an X‐linked disorder characterized by defects in cobalamin metabolism, intractable epilepsy, microcephaly, intellectual disability, movement disorders, and craniofacial abnormalities. Intractable epilepsy and neurological impairment are two of most severe phenotypes associated with cblX syndrome, however we do not understand the cellular and molecular mechanisms underlying these phenotypes in vivo. The human and mouse orthologs of HCFC1 are located on the X‐chromosome and deletion of the murine HCFC1 gene is subject to compensatory mechanisms, which have limited our ability to understand the function of HCFC1 during early brain development. To circumvent the limitations associated with rodent model systems, we created a stable germline mutation in the zebrafish HCFC1 ortholog, hcfc1a, using CRISPR/Cas9 genome editing. We identified three independent germline alleles in the hcfc1a gene, two of which result in a premature stop codon and a third that results in substitution of two amino acids within the kelch protein interaction domain. In vitro evidence suggests that perturbations in HCFC1 expression affect the number and differentiation of neural precursors. Thus, we analyzed the number of neural precursors and post‐mitotic neurons in hcfc1a mutant embryos. Mutation of hcfc1a resulted in an age dependent statistically significant increase in the number of neural precursors in specific brain regions. In addition, cell cycle analyses demonstrated a critical function of hcfc1a in the transition from S‐phase to G2/M phase. Together, our results provide evidence that HCFC1 plays an important role in the production, maintenance, proliferation, and/or differentiation of neural precursors during early vertebrate development. These data suggest a potential mechanism by which mutations in HCFC1 cause neurological deficits.Support or Funding InformationResearch reported in this poster was supported by the National Institute Of General Medical Science of the National Institutes of Health under the linked Award Numbers RL5GM118969, TL4GM118971, and UL1GM118970. 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.
Novel Regulators of Melanogenesis Identified by Genome‐wide Transcriptome Analysis
Authors: Yadav shalini , Kirty Kritika , Shukla Mudita , Natarajan Vivek , Gadgil Chetan , Gokhale Rajesh , Natarajan Krishnamurthy 
Published: June 2021

Skin pigmentation occurs through an intricate network of pathways involving several signaling and regulatory molecules. The homeostatic mechanisms controlling melanogenesis is of great importance since prolonged activation of melanin synthesis could have disastrous consequences to the skin. We used the small molecules forskolin and IBMX, two activators of cAMP levels, to induce melanogenesis in B16 melanoma cells. Treatment with forskolin and IBMX led to increased tyrosinase protein levels and enzyme activity, hyper‐dendricity, transcriptional upregulation of melanin biosynthetic pathway genes and melanin accumulation. Electron microscopic imaging showed that the two small molecules enhanced the maturation of melanosomes. To identify genome‐wide transcriptomic changes, we carried out a time‐course microarray analysis of B16 melanoma cells treated with forskolin and IBMX. Microarray data analysis showed that a large number of genes are commonly regulated by forskolin and IBMX. Hierarchical clustering analysis showed a temporal pattern of expression of ~1620 genes. DAVID pathway analysis yielded several genes involved in melanosome biogenesis, cell cycle, inflammation and small solute transporters enriched during melanogenesis. Moreover, the hyper‐pigmenting cells contained elevated ROS level consistent with the melanogenesis itself being a pro‐oxidative process indicating a connection between melanogenesis and oxidative stress in melanocytes. A large number of differentially expressed genes belonged to the SLC small molecule transporter family genes. siRNA‐mediated silencing of two of the novel transporter genes led to impaired pigmentation indicating a key role of the SLC genes. Moreover, we identified several differentially expressed genes encoding chromatin modulators and transcription factor genes including MITF, a master regulator of melanocyte lineage. Microarray profiling of B16 cells upon silencing with siMITF identified a large number of MITF‐independent genes. Together our study provided an integrated view of the transcriptome involved in melanogenesis that could facilitate the understanding of the pathogenesis of pigmentation disorders.Support or Funding InformationGenotypic Technology for microarray hybridization CSIR‐NMITLI, DST‐PURSE and DBT for fundingThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of Bio‐analytical Methodology and Physico‐chemical Characterization of Novel Compound RED
Authors: Sarkar Arindam Basu , Blanco Marsha , Reddy E Shyam P. 
Published: June 2021

ObjectiveThis study was carried out to develop the analytical methodology and limited characterization of the novel compound codenamed RED. The structure of the novel molecule is being withheld to protect the intellectual rights and patent issues involved.IntroductionThe molecule under investigation is a novel molecule that is custom manufactured for research purpose only. It is believed to have anti‐cancer activity and thus is a subject of interest and investigation.MethodThe UV‐Visble absorption spectra of the compound is established. A reverse phase liquid chromatographic method is developed to separate RED from its close structural analogs code named RmBr, RpM and RPh using a Xorbax Eclipse XDB‐C18 as stationary phase. The mobile phase is a combination of acetonitrile and water used in a continuously changing concentration gradient. Standard curve was prepared between concentrations of 0.5–10 mcg/ml with RmBr as internal standard; limit of detection (LOD) and quantitation (LOQ) was established. Statistical analysis of the data was carried out. The solubility of the compound in water, water containing 5% DMSO and water containing 5% DMSO and 5% Cremaphor was established by vortexing and ultra‐centrifugation. The stability profile of the compound in water and water containing 5% DMSO and 5% Cremaphor is established in glass and polypropylene container.ResultsWithin the specified range the standard curve yielded a slope of 0.1631 (±0.0045) and an intercept of 0.0035 (±0.0054) and good linearity (R2≥0.999). RED solubilities are found to be 0.34, 0.7 and above 5 mcg/ml in water, 5% DMSO and 5% DMSO+5% Cremaphor ( a commonly used bio‐compatible solubilizing agent) respectively. The compound is found to be stable (≥90% of original content) for more than 7, 9 and 7 days in water, 5% DMSO and 5% DMSO+5% Cremaphor respectively when preserved in room temperature in glass containers. However, the compound Red rapidly degenerates when kept in room temperature in polypropylene bottles. The shelf life (>90% of original) of Red in polypropylene containers in room temperature is less than a day. The stability studies were carried out in both transparent and amber colored glass bottles, the compound was found to stable up to 7 days. The stability is not dependent on light within the time period of investigation.ConclusionThe project is successful in establishing an analytical methodology for RED that is repeatable, robust, accurate and precise in the range. The solubility and stability of the compound in common aqueous solvents have been established. More work is required achieve a complete physico‐chemical characterization of compound.Support or Funding Information*This work has been financially supported by University of Findlay, College of Pharmacy fundsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.