bluuebunny/crossref_metadata_2025_split huggingface dataset:
Defining Functional Human Variants in the HMG Box of SOX Genes of >60,000 Human Individuals for Potential Cardiovascular and Cancer Genetic Risk
Authors: Rasicci Daniel , Underwood Adam C. , Freeland Thomas , Milsted Amy , Jacob Howard J. , Prokop Jeremy 
Published: June 2021

Our group and others have previously linked SOX (SRY‐related HMG‐box) genes to cardiovascular regulation and cancer. SOX proteins are transcription factors containing a highly conserved HMG (high mobility group) box DNA binding domain that enables sequence specific DNA interaction, bending, and gene regulation. Sequence variation in the HMG box of SOX genes has not been fully explored in humans; therefore, we have evaluated the variants within the 20 human SOX genes from ~60,000 exomes/genomes of gnomAD and also the cancer COSMIC database for somatic variations. Each variant was analyzed using codon selection and molecular modeling. A total of 3,049 variants were identified from gnomAD and 1,566 from COSMIC, with an enrichment of variants that falls within the HMG domain in cancer patients. Uniquely we highlighted an enrichment of HMG box variants in cancers of the large intestine within SOX genes with the highest enrichment in the SOX9 gene. Our most conserved and potentially impactful variant was identified as SOX18 E137K, found within 0.8% of Latino individuals. By comparing stability differences of the E137K variant against wild type SOX18 structures, using molecular dynamics simulations, SNP analysis at amino acid 137 (Glutamic acid to Lysine) was shown to disrupt the structure of the third α‐helix of the HMG box. Since other variants that fall within the HMG box of SOX18 are associated with disorders of the vasculature and lymphatics, we suspect SOX18 E137K to potentially affect vascular function through the alteration of its interaction with DNA. Work is underway to begin to assess the physiologic implications of this novel variant.Support or Funding InformationSupport for this study was from NIH‐K01ES025435 (to JWP) and Walsh UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
White light emitting diode suppresses proliferation and induces apoptosis in hippocampal neuron cells through mitochondrial cytochrome c oxydase‐mediated IGF‐1 and TNF‐α pathways
Authors: Yang Yang 
Published: June 2021

Light emitting diode (LED) light has been used in the treatment of traumatic brain injury, neural degenerative diseases and psychiatric disorders. A growing number of studies indicate that blue LED light affects cell proliferation and apoptosis in photosensitive cells and cancer cells, yet it remains unknown whether white LED light exposure impacts hippocampal neural cell proliferation and apoptosis. The present study was aimed to demonstrate the effect of white LED light exposure on viability of hippocampal neural cells and to reveal the underlying mechanisms. Cells exposed to white LED light for 24 h showed reduced viability, with higher apoptotic rate and a cell cycle arrest at G0/G1 phase. White LED light activated mitochondrial cytochrome c oxidase (Cco), in association with enhanced ATP synthase activity and elevated intracellular ATP concentration. Also, reactive oxygen species (ROS) and nitric oxide (NO) production were increased, accompanied by higher calcium concentration and lower mitochondrial membrane potential. Concurrently, the mRNA expression and the concentration of IGF‐1 were decreased, while that of TNF‐α were increased, in light‐exposed cells, which was supported by the luciferase activity of both gene promoters. The down‐stream mitogen‐activated protein kinase (MAPK), AKT/mTOR pathways were inhibited, in association with an activation of apoptotic caspase 3. N‐Acetylcysteine, a ROS scavenger, protected the cells from LED light‐induced cellular damage, with rescued cell viability and restored mRNA expression of IGF‐1 and TNF‐α. Our data demonstrate that white LED light suppresses proliferation and induces apoptosis in hippocampal neuron cells through mitochondrial Cco/ROS‐mediated IGF‐1 and TNF‐α pathways.Support or Funding InformationThe National Basic Research Program of China (2012CB124703), the Special Fund for Agro‐scientific Research in the Public Interest (201003011), the Fundamental Research Funds for the Central Universities (KYZ201212) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determining the Effect of MRP4 Knockout on Pigment Granule Position in Mouse RPE
Authors: Ibarra Dora Evelyn , Mireles Thomas , Pattillo Amanda , Przybylski Connor , Roberts Tasha , Wood Tiffany S. , Schuetz John , García Dana M. 
Published: June 2021

In the retinal pigment epithelium (RPE), pigment granules of wild type mice aggregate in darkness and disperse in light. In fish, pigment aggregation involves transport of cAMP into the RPE in vitro. We speculate that multidrug resistance protein 4 (MRP4) is responsible in vivo for the export of cAMP from the photoreceptors into the sub‐retinal space, making it available for uptake into the RPE in both fish and mice. We hypothesize if the MRP4 gene is knocked out in mice, we will not see a significant difference in pigment position between light and dark adapted mice. To test the hypothesis, we have quantified pigment position in RPE from wild type mice and knockout mice by estimating pigment granule density in the basal part of the cells examined by TEM. Bruch's membrane was used as a marker to determine the location of the base of the RPE. To measure pigment position, 2 μm × 2 μm boxes were aligned along Bruch's membrane, the pigment granules within each box were counted, and the average pigment density was calculated. The average pigment granule density for light adapted wildtype (LAWT; n = 3 mice) was 0.7 pigment granules/μm2 (± 0.2 pigment granules/μm2) and for dark adapted wildtype (DAWT; n = 3 mice) was 1.4 pigment granules/μm2 (± 0.3 pigment granules/μm2); pigment density was statistically significantly different (P < 0.02) between LAWT and DAWT. Preliminary data suggests the average pigment granule density for dark adapted MRP4 knockout is 0.57 pigment granules/μm2 (DAKO; n = 2). Our results with WT indicate basal pigment density increases in the dark, consistent with pigment aggregation and suggests dark adaptive pigment aggregation may be impaired in knockout mice.Support or Funding InformationThis project was supported by an Undergraduate Research Fellowship to AP and funds from the Office of the Provost awarded to DG in recognition of her service as Presidential Fellow. DEI received salary support from HSI STEM Program (84.031c) Award #P031C160035.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 Inositol hexakisphosphate Kinase 1 (IP6K1) Does Not Increase Akt or mTOR Activity in vitro.
Authors: Barclay Richie , Mackenzie Richard 
Published: June 2021

BackgroundInositol hexakisphosphate Kinase 1 (IP6K1) has been identified as a key signalling enzyme that inhibits Akt activity in hepatic cell lines and diabetic rodent models (Chakraborty et al., 2010; 2012; Goshal et al., 2016). Given both glucose transport and protein synthesis require Akt activation, we investigate if IP6K1 altered the IGF ‐ Akt – mTOR signalling cascade.MethodsC2C12 mouse skeletal myoblast cells were cultured in 12mL growth media (500mL DMEM, 10% FBS, 1% anti ‐ anti, 1% penicillin ‐ streptomycin) under standard conditions, changing media every 24 – 48 hours. Once 80% confluence was reached myoblasts were incubated in 12mL differentiation media (500mL DMEM, 2% donor equine serum, 1% anti ‐anti, 1% penicillin ‐ streptomycin) for 96 hours, changing media every 24 – 48 hours, to form mature myotubes. Myotubes were treated with IGF −1 (10ng/ml) +/− N2‐(m‐(trifluoromethy)lbenzyl) N6‐(p‐nitrobenzyl)purine (TNP; 10μm) for 24 hours before being lysed (1X cell lysis buffer supplemented with protease and phosphatase inhibitor cocktail) and total protein content determined using Lowry method. Western blot analysis was used to quantify t‐Akt, p‐Akt308, p‐Akt473, t‐mTOR, p‐mTOR2448 and IP6K1.ResultsA significant condition x IP6K1 effect was noted (P < 0.05) between IGF – 1 treatment and control in C2C12 myotubes, whilst IGF – 1 + TNP decreased IP6K1 content compared to IGF – 1 alone (P < 0.05). Significant differences were observed between IGF – 1 + TNP and control in p‐Akt308 and p‐Akt473 (P < 0.05), however no significant differences were observed between IGF – 1 + TNP and control in p/Akt308, p/Akt473 and p/mTOR2448 (P > 0.05).ConclusionsThis research is the first to characterize IP6K1 signalling in the IGF – 1, Akt – mTOR signalling cascade in C2C12 muscle cells. A reduction in IP6K1 did not increase p‐Akt or p‐mTOR activity.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modulating Cytoskeletal Structure And Cellular Signaling To Target Neuron Cell Membrane Repair
Authors: Paleo Brian J. , Bhattacharya Sayak , Weisleder Noah 
Published: June 2021

Most cells in the human body have the capacity to reseal their cellular membranes following disruption of the lipid bilayer. The cell signaling response regulating membrane repair is dependent on changes in the mechanical tension on the plasma membrane. Increased mechanical tension on the membrane can lead to membrane disruptions remaining open and thus preventing resealing of the membrane. Disruption of these membrane repair processes can result in a number of diseases including muscular dystrophy, Alzheimer's and neurodegeneration. Conversely, if membrane repair can be elevated in striated muscle or neuronal tissues, this response could potentially minimize cell death in neuromuscular diseases and alleviate the progression of these disease states. We have previously shown that tripartite motif (TRIM) 72, also known as mitsugumin 53 (MG53), can increase plasma membrane repair in skeletal and cardiac muscle and in non‐muscle cell types where it is not usually expressed. Our recent results indicate that there are distinct TRIM72/MG53 regulated cellular signaling events that control the membrane repair response in multiple cell types. This observation led us to screen for novel TRIM family proteins that may be able to mediate membrane repair in neuronal cells. We found that TRIM2 transfected cells show an increased capacity for membrane resealing following multi‐photon laser injury and that knockdown of TRIM2 decreases membrane repair. Because TRIM2 is highly expressed in the nervous system and was previously shown to regulate neurofilaments (NFL), the objective of this study is to prove that TRIM2 protects against membrane damage in neurons through regulation of neurofilament ubiquitination and that disruption of TRIM2 or NFL leads to compromised membrane repair and neuron cell death. Through confocal microscopy, we have shown that TRIM2 co‐localizes with NFL in neurons. Following injury to the cell this association is altered, leading to changes in the subcellular localization of TRIM2 and an apparent reorganization of the neurofilament cytoskeleton. Given the extensive expression of TRIM2 in the nervous system, it may represent a good candidate for targeting membrane repair in the treatment of a variety of neurodegenerative diseases.Support or Funding InformationAmerican Heart Association, National Institutes of Health, National Institute of Neurological Disorders and Stroke (NINDS)‐sponsored T32, and Jain FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Damage Response in Doxorubicin‐Exposed Cardiac Fibroblasts
Authors: Mancilla Trevi Ann , Aune Greg 
Published: June 2021

Pediatric cancers survivors exposed to doxorubicin during treatment are at an increased risk for cardiovascular disease. This study investigates two known responses to doxorubicin injury, the p53‐DNA‐damage pathway and the Parkin‐mitophagy pathway in the cardiac fibroblast. Doxorubicin is known to upregulate p53 expression. However, mitophagy of the mitochondria damaged by doxorubicin may be inhibited by this upregulation. Previous studies in cardiac myocytes have shown that cytosolic p53 can bind to Parkin and prevent it from localizing to damaged mitochondria, an early step of mitophagy. This study investigated if this also occurs in cardiac fibroblasts. Expression levels of p53 and Parkin were measured in primary cardiac fibroblasts exposed to doxorubicin. Immunoblotting and fluorescent microscopy both demonstrated increases in p53 and Parkin after doxorubicin exposure (p<0.05). Furthermore, the pattern of Parkin staining remains diffuse throughout the cytoplasm of the doxorubicin‐treated cells. Fibroblasts treated with a mitochondrial uncoupler had a linear staining pattern that may be due to the localization of Parkin with the mitochondria. As p53 is not upregulated by the uncoupler, Parkin localization would not inhibited. Future studies will determine if doxorubicin‐induced upregulation of p53 inhibits mitophagy and the effect this has on the cardiac fibroblast phenotype. Since cardiac fibroblasts mediate adverse cardiac remodeling, stress‐induced phenotypic changes to the cell will impact this process. Therefore, effects on the cardiac fibroblast are essential to understanding the development of the dilated cardiomyopathy seen in these patients.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Improved cardiac remodeling after in vivo knockdown of type VI collagen following myocardial infarction
Authors: Baker Nicholas J. , Minasyan Ashot , Adapala Ravi , McShannic Christian M. , Gan Connie , Hom Brian L. , Thodeti Charles K. , Meszaros J. Gary 
Published: June 2021

RationaleOur previous studies have shown that hearts of Col6−/− mice are protected from myocardial infarction (MI) injury, as these hearts exhibit reduced myocyte apoptosis, fibrosis and wall thinning leading to preserved long‐term cardiac function. The current project was designed to test whether knockdown of Col6 is feasible in the rat MI model and carries a similar cardioprotective effect during post‐MI remodeling.ObjectiveTo utilize a novel, non‐viral delivery method to deliver and determine whether siRNA‐mediated knockdown of Col6 is effective in the infarcted rat heart and affords cardioprotection in vivo in the weeks following MI injury.Methods and ResultsMI was induced via ligation of the left anterior descending artery (LAD) in adult Sprague‐Dawley rats and cardiac function was measured by transthoracic echocardiography. Calculated changes in endocardial volume, percent ejection fraction (%EF), and percent fractional shortening (%FAC) were performed at 1–6 weeks post‐MI. We witnessed a decrease in mean endocardial %EF of 44.7 ± 1.4 % in vehicle treated (control) rats, a parameter that was higher in siRNA treated hearts at 47.4 ± 1.8 %. A similar improvement was also seen in endocardial %FAC, which was 30.1 ± 1.4 % in vehicle treated rats versus 34.0 ± 1.3 % in siRNA treated rats. Knockdown of Col6 in vivo resulted in lower endocardial diastolic volume in siRNA treated rats (654.1 ± 81.3 μl in the vehicle group compared to 549.8 ± 8.4 μl for the siRNA treated group) and lower systolic volume in siRNA treated rats (362.7 ± 49.9 μl in the vehicle group compared to 291.6 ± 9.6 μl in the siRNA treated group).ConclusionssiRNA mediated Col6 knockdown using JET‐PEI as a novel delivery method was effective in vivo, reducing Col6 expression levels over 75% in the anterior wall of the LV. Echocardiography revealed that the siRNA injections preserved cardiac function and prevented left ventricular chamber dilation following MI, indicating that this approach may have a positive therapeutic benefit in the recovery from MI injury.Support or Funding InformationNIH R15HL132312This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
AMP‐Activated Protein Kinase (AMPK) Modulates Myoendothelial Junctions (MEJ) in Murine Microvessels
Authors: Qiu Jiehua , Tahir Sibgha , Schubert Kai‐Michael , Schneider Holger , Pohl Ulrich 
Published: June 2021

Myoendothelial junctions (MEJ) in microvessels are not only the site of direct cell communication via gap junctions but also represent a signaling microdomain critical for regulating vascular processes such as endothelium dependent hyperpolarization (EDH). The 5′‐adenosine monophosphate‐activated protein kinase (AMPK) has been shown to influence vascular tone. It has not yet been studied, however, whether AMPK could also affect MEJ. Thus, the aim of this project was to elucidate the impact of AMPK on MEJ dynamics and its potential mechanism of action.We studied isolated small mesenteric and skeletal muscle resistance arteries of mice using a pressure myograph system developed in our laboratory. Using confocal and two photon excitation fluorescence microscopy we identified the internal elastic lamina, which in these vessels exhibits small holes that allow direct contact between endothelial and smooth muscle cells. On average, we found that in about 35% of the holes, structures with a bright actin core as assessed by using F‐actin fluorescence in LifeAct mice or by staining with phalloidin which we could define as sites of MEJ. These structures showed expression of Cx37 and Cx43 suggesting that gap junctions (MEGJs) were also localized in these structures. Over observation times of up to 2 hours, we found that the number and localization of these MEJ varied, while they always positioned in the area of holes of IEL only. Thus, for the first time we could investigate MEJ dynamics. AMPK negatively regulates MEJ formation, since in AMPK α1, but not α2 knockout (KO) mice, the number of MEJ was significantly increased. In accordance, in wild type mice, incubation with the AMPK inhibitory compound C (CC) significantly augmented the number of MEJ. Furthermore, KO of AMPK α1, in intact mice as well as in cultured human and porcine smooth muscle cells, went along with an increase in PAI‐1 expression. Accordingly, incubation of arteries with exogenous PAI‐1 also increased the number of MEJ. The negative effect of AMPK on PAI‐1 could be explained by enhanced expression of the silencer of the PAI‐1 gene, small heterodimer partner (SHP). Since MEJ are also involved in EDH, we studied whether an enhanced formation of MEJ correlated with the magnitude of EDH induced vasodilation. Indeed, a higher amount of MEJs in α1 KO mice went along with a left shift of acetylcholine (ACh) ‐induced dilation dose effect curve.This work describes for the first time a role of the α1 subunit of AMPK as a potent modulator of MEJ dynamics which is probably mediated by controlling PAI‐1 expression. Our data also for the first time demonstrate a dynamic regulation of MEJ expression in intact blood vessels.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Envigo's Female Salt‐Sensitive Rapp Rats are now Spontaneously Hypertensive and have Higher Frequencies of CD4+, CD4+ CD25+ and CD4+ CD25+ FoxP3+ T cells compared to Normotensive Salt‐resistant Rapp Rats
Authors: Pai Amrita V. , West Crystal A. , Souza Aline , Kadam Parnika S. , Pollner Emma J. , West David A. , Ji Hong , Wu Xie S. , Baylis Chris , Sandberg Kathryn 
Published: June 2021

IntroductionWe and others have found female Rapp (Jr) salt‐sensitive (SS) rats recently purchased from Envigo, formerly Harlan (Hsd) and maintained on a low sodium diet are spontaneously hypertensive. To further investigate the role of T cells in the new phenotype of spontaneous hypertension (SH) observed in the Envigo's Rapp strain (SHSS/JrHsd), we compared the T cell profile in SHSS/JrHsd with that of the Rapp normotensive salt‐resistant rats (NSR/JrHsd).MethodsMean arterial pressure (MAP) was measured by radio‐telemetry in juvenile (j) 1 month old (mo) SHSS/JrHsd, and in young (y) adult 4 mo SHSS/JrHsd and NSR/JrHsd rats. Helper (CD4+) and cytotoxic (CD8+) T cells were assessed in the kidney by multicolor flow cytometry in 1 and 4 mo SHSS/JrHsd and NSR/JrHsd rats.ResultsRadio telemetry confirmed that 1 mo jSHSS/JrHsd rats had normal MAP [(mmHg): 100±1; n=13]. Blood pressure rapidly increased with age and was significantly higher in 4 mo ySHSS/JrHsd rats compared to age matched normotensive yNSR/JrHsd [(mmHg): yNSR/JrHsd, 103±2 vs. ySHSS/JrHsd, 160±11; p<0.007; n=3/group]. The frequencies of CD4+ (1.3 fold; p<0.0001), CD4+ CD25+ (1.5 fold; p=0.002) T cells and the CD4+/CD8+ ratio [(CD4+/CD8+): jNSR/JrHsd, 1.1±0.07 vs. jSHSS/JrHsd, 2.1±0.1; p<0.0001; n=8/group] were significantly higher in jSHSS/JrHsd compared to jNSR/JrHsd. The frequencies of CD4+ (1.4 fold; p<0.0001), CD4+CD25+ (2.1 fold; p<0.0001) T cells and the CD4+/CD8+ ratio [(CD4+/CD8+): yNSR/JrHsd, 1.6±0.07 vs. ySHSS/JrHsd, 5.6±0.6; p<0.0001; n=7–8/group] were even higher in ySHSS/JrHsd compared to yNSR/JrHsd. Moreover, the ySHSS/JrHsd rats had a higher frequency of CD4+CD25+ FoxP3+ T cells compared to the yNSR/JrHsd at 4 mo (2 fold; p=0.0009).ConclusionsFemale Rapp SS rats from Envigo (formerly Harlan; Hsd) are normotensive as juveniles but get spontaneous hypertension by the time they are 4 mo; they should more appropriately be referred to as SHSS/JrHsd Rapp rats. Our T cell findings support previous studies indicating CD4+ T cells and subtypes contribute to hypertension.Support or Funding InformationTL1‐TR001431 (AVP), UL1‐TR001409 (KS) and R01‐HL119380 (KS, HJ)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.