bluuebunny/crossref_metadata_2025_split huggingface dataset:
Perforasomes of the facial artery
Authors: Hirtler Lena , Berger Vinzenz 
Published: April 2020

BackgroundThe facial artery perforator flap is a valuable alternative for reconstruction of small to medium defects in the central midface. Not only does it provide pleasing cosmetic results, it also allows reconstruction in a one‐stage procedure. Even though facial artery perforators have been examined by anatomical studies during the past years, there is still little knowledge of the vascular territories provided by these vessels. Aiming to make facial artery perforator flaps a plannable and safer procedure, this study examines the location and size of the vascular territories provided by facial artery perforators.Material and methods20 facial arteries of fresh specimen have been dissected. All perforators with an external diameter of at least 0,50 mm have been identified and injected with Methylene blue‐ or Eosin Y‐solutions to color their appendant vascular territories. All colored skin territories have been photographed, digitalized and measured. Their location, as well as the location of the perforating vessels, has been shown. All results were statistically analyzed.Results95 perforators with an external diameter of at least 0,50 mm have been identified. The average number of perforators per facial artery was 4,75. The mean external diameter at the origin of the facial artery was 0,64 ± 0,14 mm. The mean size of the colored skin territories was 5,62 ± 4,83 cm2. There was a statistical correlation between perforator diameter and the size of its vascular territory.ConclusionTwo areas have been shown in which facial artery perforators originate frequently. Their location has been described in relation to a bone structure. Size and shape of the vascular territories show a high variability with regions of increased density. The graphical illustration of the location of the facial artery perforators and their vascular territories can help making facial artery perforator flaps a better plannable procedure.
Ppargc1a Regulates Prostaglandin Signaling to Control Ciliogenesis and Renal Multiciliated Cell Fate Choice During Development
Authors: Chambers Joseph M. , Addiego Amanda , Wingert Rebecca A. 
Published: April 2020

Cilia are microtubule‐based organelles that function in a multitude of physiological contexts to perform chemosensing, mechanosensing, or fluid propulsion. The process of ciliogenesis is highly regulated and disruptions result in disease states termed ciliopathies. The genetic and molecular events that lead to dysfunctional ciliated cells are still not fully understood. Here, we show novel roles for peroxisome proliferator‐activated receptor gamma 1 alpha (ppargc1a) during ciliogenesis in mono‐ and multiciliated cells (MCCs) as well as discernment of renal tubule MCC fate choice during embryogenesis. We discovered that ppargc1a performs both roles by affecting prostaglandin levels, where cilia formation and renal MCC fate were restored with prostaglandin E2 (PGE2) treatment. Genetic disruption of ppargc1a specifically reduced expression of the prostanoid biosynthesis gene prostaglandin‐endoperoxide synthase 1 (ptgs1), and suboptimal knockdown of both genes revealed a synergistic effect. Further, ptgs1 overexpression rescued ciliogenesis and renal MCCs in ppargc1a deficient embryos. These findings position Ppargc1a as an essential genetic regulator of prostaglandin synthesis during ciliated cell ontogeny.Support or Funding InformationNational Institutes of Health (R01DK100237 to R.A.W.), a 2019 University of Notre Dame Advanced Diagnostics and Therapeutics Graduate Fellowship Award (to J.M.C.), and a 2019 University of Notre Dame College of Science Summer Undergraduate Research Fellowship Award (to A.A.)
Investigating the role of a V‐ATPase in the Development of the Air Sac Primordia
Authors: Salim Usman , Salim Hasan , Gondal Anas , Srivastava Ajay 
Published: April 2020

Drosophila melanogaster is a commonly used organism in biological research. Their Air Sac Primordium (ASP) is analogous to the human lungs in that it supplies oxygen to the flight muscles. Furthermore, the ASP has emerged as a good structure to study cellular invasive processes with the potential to aid in our understanding of tumor metastasis. The ASP originates on the outside of the wing imaginal disc and then burrows into the wing imaginal disc as it develops. What genes are involved in this process is not well understood. A genetic screen for genes that are expressed in the ASP identified a proton pump gene encoding a V‐ATPase. The V‐ATPase is a vacuolar proton pump that plays a large array of roles from regulating the pH of cells internal space, to regulating endocytosis, to processing proteins and degradation. In insects, the V‐ATPase are important in establishing a membrane potential to drive K+ secretion. It has also been found to influence tracheal branching. Using RNA interference, the gene for the V‐ATPase was knocked down resulting in a defect in ASP development. Data from these and other experiments will be presented and their significance will be discussed.Support or Funding InformationKentucky Biomedical Research Infrastructure Network (KBRIN) GrantGatton Academy Research Internship Grant (RIG)Faculty Undergraduate Student Engagement (FUSE) Grant
Transient ischemic stroke induces substantial leukocyte recruitment to both the ischemic and non‐ischemic cerebrovasculature as visualized through live animal imaging.
Authors: Batra Ayush , Nadkarni Neil A. , Muller William A. , Sullivan David P. 
Published: April 2020

Ischemic stroke is a leading cause of a death and disability worldwide. Current treatment strategies rely on early restoration of blood flow; however, injury persists despite successful reperfusion. The innate immune response plays a pivotal role in reperfusion injury, but the mechanisms and time‐course of neutrophil and monocyte recruitment to the cerebrovasculature (CBV) during ischemic stroke remain unclear. We sought to characterize the real‐time leukocyte response at the CBV in a mouse stroke model with reperfusion.To visualize leukocyte recruitment induced by ischemia, Ly6G‐tdTomato mice (which have fluorescent red neutrophils) were subjected to 90 minutes of transient middle cerebral artery occlusion (tMCAO) followed by reperfusion. Real‐time spinning‐disc confocal intravital microscopy of the CBV was performed at 24 and 72 hours following reperfusion. Intracranial windows were placed in both ischemic and non‐ischemic hemispheres; and tMCAO treated mice were compared to mice that underwent sham surgery. Neutrophil recruitment (adhesion, rolling) and extravasation were quantified in vivo on both the ischemic and contralateral non‐ischemic hemispheres. Postmortem staining with 2,3,5‐triphenyltetrazolium chloride was used to confirm induction of the stroke and to quantify infarct volume. In addition, wide‐field microscopy was used to examine neutrophil extravasation across the parenchyma.Animals subjected to sham surgery showed negligible leukocyte recruitment to the CBV. However, marked increases in leukocyte rolling, adhesion, and extravasation were observed at 24 and 72 hours following the tMCAO procedure. Interestingly, non‐ischemic hemispheres also demonstrated increased leukocyte rolling and adhesion, but not extravasation tMCAO group. Also, substantially more neutrophil extravasation was observed in the core of the infarct at 72 hours. These findings are consistent with an early recruitment of neutrophils to the cortical surface at early time points and a delayed infiltration into the core, as reported by others.These results show that a significant leukocyte response persists after reperfusion following tMCAO. This response is present both on the ischemic and, to a lesser extent, the non‐ischemic cortical CBV. Acute leukocyte recruitment observed on the non‐ischemic hemisphere suggests that ischemic stroke triggers a global inflammatory response within the CBV. Further reducing inflammation through immunomodulatory therapy in conjunction with reperfusion therapy may be beneficial in not only reducing stroke volume but potentially mitigating inflammation mediated secondary neuronal damage.Support or Funding InformationNIH R01 HL046849 and R37 HL064774 to W.A.M.
Characterization and Quantification of Dopaminergic Phenotypes via Computer‐aided Video Analysis
Authors: Garriga Gustavo Adolfo , Nelms Brian 
Published: April 2020

Dopamine is an important neuromodulator known to regulate a myriad of brain functions ranging from motor control to memory and learning. Dopamine dysregulation has been associated with multiple neurological disorders including schizophrenia, attention deficit hyperactivity disorder, addiction, Parkinson’s disease among many others. To better understand dopamine function at the molecular level, we are investigating the roles and regulation of dopaminergic genes using Caenorhabditis elegans (C. elegans) as a model system. We are conducting swimming induced paralysis (SWIP) assays using a computer‐aided video analysis system, allowing us to collect more nuanced data to uncover new phenotypes and discover roles for genes involved in dopaminergic signaling. We have selected several genes to focus on in this study. Each of these genes is highly enriched in dopamine neurons, based on RNASeq data from our laboratory. Mutant strains for these genes display accelerated swimming‐induced paralysis, a phenotype characteristic of hyperdopaminergic signaling, when compared to the wild‐type strain, N2. Via this analysis, we propose to gain more insight into specific rates and patterns of paralysis of these mutant strains in order to identify novel genes critical for dopaminergic functioning.Support or Funding InformationFunding support from NSF CREST Grant #HRD1547757, NIH BD2K R25 Grant #1R25MD010396‐01, and DOE Title VII Grant MD‐HBCU #P382G090004.
Influence of RGS6 on Inhibitory G Protein Signaling in Mouse Sinoatrial Nodal Cells
Authors: Anderson Allison , Masuho Ikuo , Marron Ezequiel , Martemyanov Kirill , Wickman Kevin 
Published: April 2020

IntroductionM2 muscarinic (M2R) activation of G protein‐gated inwardly rectifying K+ (GIRK) channels in sinoatrial nodal (SAN) cells is a key mediator of vagal/parasympathetic slowing of heart rate. The strength of M2R‐GIRK signaling is negatively regulated by Regulator of G protein Signaling 6 (RGS6). Adenosine, used clinically to diagnose and treat certain arrhythmias, exerts these actions through A1 adenosine receptor (A1R) activation of GIRK channels. The extent to which RGS6 regulates A1R‐GIRK signaling, however, is unclear. Here, we investigated the impact of RGS6 on A1R‐GIRK and M2R‐GIRK signaling in mouse SAN cells.Methods and ResultsWe measured the efficacy and potency of M2R‐ and A1R‐GIRK signaling in adult SAN cells from wild‐type and RGS6−/− mice using whole‐cell patch‐clamp electrophysiology. As previously reported, SAN cells from RGS6−/− mice displayed prolonged channel deactivation kinetics and increased channel sensitivity to the non‐selective cholinergic agonist carbachol (CCh), relative to wild‐type counterparts. In contrast, we observed a striking increase in the amplitude of the GIRK response evoked by adenosine (Ado) in RGS6−/− SAN cells compared to wild‐type controls, with no impact on channel deactivation kinetics or sensitivity. Further recordings in wild‐type SAN cells revealed that M2R activation completely occludes the Ado‐induced response, whereas Ado application activates only a smaller fraction of GIRK channels. Both CCh‐ and Ado‐ induced currents were abolished in SAN cells treated by pertussis toxin (PTX), indicating these responses are mediated by Gi/o G proteins. Utilizing a bioluminescence resonance energy transfer (BRET) assay in transfected HEK cells, we found that RGS6 prefers Gαo as a substrate for over Gαi. Additionally, we found that while A1R does not discriminate between Gαo and Gαi, M2R displays a clear preference for Gαo. We next employed a Cre/flox strategy to remove Gαo from atrial tissue, including SAN cells. Loss of Gαo yielded significantly blunted CCh‐induced responses, which also displayed prolonged activation and deactivation kinetics. While Ado‐induced responses had prolonged deactivation kinetics, activation kinetics and response amplitude were unaffected.ConclusionsThese results suggest that RGS6 negatively regulates A1R‐GIRK signaling distinct from its influence on M2R‐GIRK signaling in mouse SAN cells likely due to differing utilization of Gao by M2R and A1R.Support or Funding InformationThis work was supported by NIH grants R01 HL105550 and F31 HL139090.
Exploring The Relationship Between Optimized Bioavailable Curcumin And Performance After an Intensified Training Program
Authors: Kisiolek Jacob N. , Flores Victoria , Ramani Arjun , Johnston Nora , Taylor Sophia , Kheradia Nikeeta , Stewart Laura K. 
Published: April 2020

ObjectiveThe purpose of this double blind, randomized, study was to determine the effects of Longvida® Optimized Curcumin on time trial performance time as well as the inflammatory mediator C‐reactive protein (CRP) after 2 weeks of an intensified training program.MethodsHealthy men and women (aged 18–40 y) were randomly assigned to placebo (PLA; n=11), Curcumin 1 (LV1; n=12) or Curcumin 2 (LV2; n=11) groups. Participants consumed capsules of rice flour placebo (1000mg/day) or curcumin (1000mg/day; Longvida 1 (LV1) or 1000mg/day Longvida 2 (LV2)) for 14 days, during a 2‐week intensified, cycle training period, which consisted of 3 training sessions per week. Each training session consisted of 6, 90 second stationary cycling sprints at 80–90% heart rate max (HRmax) followed by 180 seconds of low intensity cycling at 50–60% HRmax. Age, body mass index (BMI), percent body fat (BF%; BodPod, Cosmed, USA), and peak oxygen uptake (VO2peak) were assessed at the pre intervention time point (PRE). Time trial performance (16.1 km; TT) was obtained on a stationary cycle and resting and fasted serum was evaluated for CRP using an enzyme‐linked immunosorbent assay at both PRE and post intervention (POST) time points. Differences among groups were assessed using a one‐way ANOVA, and PRE and POST measurements among groups were assessed using a two‐way mixed model ANOVA. Data are presented as mean ± standard deviation with significance set at α=0.05.ResultsAt PRE, there were no significant differences between groups with respect to mean participant characteristics (age: 24.8±4.2 y; BMI: 24.0±3.5; BF%: 20.55±8.90; VO2peak 42.07±8.90 ml/kg/min). TT performance significantly improved an average of 1.41±5.01% (p=0.05) when PRE times were compared to POST. Additionally, although not statistically significant, the average TT time change from PRE to POST was −205.67 sec for PLA, −2.17 sec for LV1, −105.32 sec for LV2. There were no significant group effects or group by time interactions for CRP (PLA: 3.2 ±3.4 mg/L; LV1: 0.7 ±0.9 mg/L; LV2: 1.2±01.3 mg/L).ConclusionsThe intensified training period elicited a significant improvement in TT performance. When taken daily, the dose of curcumin used in this study did not appear to provide additional performance related benefits. However, when compared to the existing literature, the dose and the timing of curcumin intake as well as the mode of the activity may be related to curcumin‐mediated improvements in performance.Support or Funding InformationVerdure Sciences, Noblesville, IN
The E3 Ubiquitin Ligase Adaptor FBXL13 Mediates the cGMP‐dependent NKCC2 Ubiquitination in Thick Ascending Limbs
Authors: Ares Gustavo 
Published: April 2020

NaCl reabsorption by the thick ascending limb (TAL) is mediated by the Na/K/2Cl cotransporter (NKCC2). Nitric Oxide and atrial natriuretic peptide decrease NaCl absorption in the TAL by increasing the second messenger cyclic guanosine monophosphate (cGMP). NKCC2 is constitutively ubiquitinated, and cGMP enhances the rate of ubiquitination. Ubiquitination is a post‐translational modification that regulates protein activity or fate. We recently showed that the cGMP‐dependent NKCC2 ubiquitination is mediated by the SCF (Skp1‐Cullin1‐FBox) complex, which is the largest family of E3 ubiquitin ligase in mammals. To study new proteins that interact and regulate NKCC2, we used a targeted proteomics approach. TALs were incubated with GST constructs of the NH2‐ or COOH‐terminal portion of NKCC2 followed by pull‐down and electrophoresis. We identified the ubiquitin‐ligase adaptor F‐Box leucine‐rich repeat 13 (FBXL13) that bind the NH2‐terminus portion of NKCC2. A band at the expected molecular weight for FBXL13 confirmed FBXL13 expression in rats TALs by Western blot. We studied the interaction of full length NKCC2 with FBXL13 and found that FBXL13 co‐immunoprecipitated with full length NKCC2 in TAL lysates Therefore, we hypothesized that the cGMP‐dependent NKCC2 ubiquitination is mediated by FBXL13. To test our hypothesis, we generated a global FBXL13 knockout mice by crossing FBXL13‐Flox with a global Cre mice. We confirmed the deletion of FBXL13 in tail by PCR and in TALs by Western blot. First, we studied whether deletion of FBXL13 affects total NKCC2 expression. For this, TALs from C57BL6J and FBXL13‐KO mice were lysed, and equal amount of proteins resolved by SDS‐PAGE and detected by Western blot. We found that total NKCC2 expression in FBXL13‐KO mice is increased by 73% (C57BL6J: 100% vs. FBXL13‐KO mice: 173 ± 37%, n = 4, p<0.05). These data suggest that in FBXL13‐KO mice, NKCC2 is been accumulated and not degraded. We tested whether the cGMP‐induced NKCC2 ubiquitination is blunted in FBXL13‐KO mice. TALs from C57BL6J control mice and FBXL13‐KO mice were incubated at 37°C during proteasome inhibition with MG132 and treated with vehicle or db‐cGMP 500μM. Ubiquitinated proteins were pulled down and NKCC2 measured by Western blot. We found that cGMP increased ubiquitinated‐NKCC2 by 24 ± 10% in C57BL6J mice, whereas this effect was completely absent in FBXL13‐KO mice (−14 ± 12%, n=3, p<0.05). These data indicate that FBXL13 mediates the cGMP‐dependent NKCC2 ubiquitination in native TALs. We conclude that the cGMP‐dependent NKCC2 ubiquitination is mediated by the novel E3 ubiquitin ligase adaptor FBXL13. To our knowledge, this is the first evidence showing an E3 ubiquitin ligase that interacts with the NH2‐terminus portion of NKCC2 in native TALs.Support or Funding InformationHenry Ford Health System, Institutional Support, Grant #A20042.
Salinomycin and its derivatives as potent RET transcriptional inhibitors for the treatment of medullary thyroid carcinoma (MTC).
Authors: Alqahtani Tariq M. , Sun Daekyu M. 
Published: April 2020

Rearranged during transfection kinase (RET) is a validated molecular target in medullary thyroid cancer (MTC), as activating mutations in RET are often associated with the development of MTC. The present study reports the first preclinical characterization of salinomycin and selected analogs as potent RET transcriptional inhibitors. Reverse transcription‐PCR and immunoblotting revealed that salinomycin profoundly decreased RET expression in the TT human MTC cell line by inhibiting RET transcription. Moreover, salinomycin resulted in remarkable anti‐proliferative activity against MTC that is driven by RET (gain of function mutation) by selectively inhibiting the intracellular PI3K/Akt/mTOR signaling pathway. Also, flow cytometry and fluorescence‐activated cell sorting showed that salinomycin induces G1 phase arrest and apoptosis by reducing the expression of retinoblastoma protein, E2F1, cyclin D and CDK4. The structure‐activity relationship of salinomycin was investigated in this study. Some of the salinomycin derivatives showed the ability to reduce RET expression where others fail to alter RET expression. These results suggest that the RET‐suppressing effect of salinomycin may be largely attributed to disruption of the Wnt pathway, presumably through interference with the ternary LRP6‐Frizzled‐Wnt complex. Furthermore, these findings support the further preclinical evaluation of salinomycin and its analogs as a promising new class of therapeutic agents for the improved treatment of MTCSupport or Funding InformationAmerican Cancer Society for their support of this study (grant no. RSGM‐12‐046‐01‐CDD).
3D Virtual Models to Visualize Pelvic Floor & Perineum in Medical Gross Anatomy
Authors: Ismat Karim Hussain , Afifi Linda , Zumwalt Ann 
Published: April 2020

IntroductionThe pelvic floor and perineum are difficult regions to visualize with most educational tools. Cadaveric dissection of this region is cumbersome due to limited accessibility and ambiguity of structures, and presents challenges with teaching spatial orientation. Supplemental learning material such as two‐dimensional anatomical illustrations often lack the ability to demonstrate spatial depth and complex relationships of the pelvis and perineum. We developed 3D virtual models to teach this spatially complex region in a didactic session on pelvic and perineal anatomy.AimThis study explored the efficacy of 3D virtual models as a supplementary teaching tool for pelvic floor and perineum.MethodsTwo review sessions were provided to a total of 37 first year medical students. The content of the lesson was initially presented in a traditional lecture format alongside 2D illustrations and schematics. Immediately following the lecture portion, 3D models of each region of interest were presented to students using 3D4Medical Complete Anatomy application.Prior to the session, students (n=31) completed a modified Likert scale (1–5) survey on their familiarity with pelvic musculature, perineal fasciae and spaces, and the course of the pudendal nerve. After the session, students (n=37) completed a similar survey reviewing their confidence in pelvic musculature, perineal fasciae and spaces, and the course of the pudendal nerve. Relative frequencies for each question have been reported as a percentage, with a score of 3 considered neutral.ResultsOf the students who responded to the pre‐session survey (n=31), 6.5% reported being familiar (a score of 4 or 5) with the muscles of the pelvic floor, 9.7% reported being familiar with the perineal fasciae and spaces, and 12.9% reported being familiar with the course of the pudendal nerve.Responses to the post‐session survey (n=37), revealed that 91.9% of students reported improved confidence (a score of 4 or 5) in their understanding of pelvic floor musculature and perineal fasciae and spaces. While, 83.8% reported improved confidence in their understanding of the pudendal nerve course.ConclusionThe use of 3D models, as a supplement to existing course content, is an effective method for improving student confidence in pelvic floor and perineum anatomy. Our results suggest that the combination of 3D virtual models and didactic knowledge transmission, facilitates the learning of pelvic floor and perineal regions.