bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mast Cell Concentrations Correlate with Hepatic Fibrosis in Cancer Patients
Authors: Lombardo Jamie Lynne , Taggart Melissa , Rashid Asif 
Published: April 2020

BackgroundThe pathogenesis of liver fibrosis in non‐alcoholic steatohepatitis (NASH) has been studied in animal models and humans, with some researchers finding a direct correlation between the concentration of mast cells and the stage of fibrosis in NASH. This suggests that mast cells play a key role in the progression of fibrosis in NASH patients.This study investigates patients with liver fibrosis in a more varied clinical setting with exposure to hepatotoxic chemotherapy regimens and checkpoint inhibitor therapy with various underlying causes of hepatic fibrosis including alcohol use, chronic hepatitis C or Hepatitis B infection, and NASH. The goal is to determine if correlations between mast cell concentration (MCC) and fibrosis remain in patients with multifactorial liver injury.DesignHepatic tissue from seventy‐seven patients (13 resections and 64 biopsies), was retrospectively selected from the database of a large cancer center, and reviewed. Of the cases selected, 38 were male and 39 were female. The gender breakdown by stage of fibrosis is detailed in figure
1. Patients ranged in age from 28–87. The stage of fibrosis was determined using the Batts‐Ludwig criteria. Potential injury‐inducing factors including chemotherapy regimen, checkpoint inhibitor therapy use, and any background liver disease were noted. Six of the patients had multiple primary cancers, 47 had cancer plus underlying liver disease, 18 had liver disease alone, 11 were taking checkpoint inhibitors, 30 were on chemotherapy, 7 were taking hormone therapy, and 6 had one primary cancer with no liver involvement or underlying liver disease.A tryptase immunohistochemical stain (clone AA1, Dako) was performed on a representative section from each case to evaluate MCC. Mast cells in five 40× fields (1 mm2) in both portal tracts and lobular areas were counted and separately recorded. MCC was then compared to the stage of fibrosis using an independent student’s t‐test calculation (Graphpad Prism statistics software).ResultsMCC in portal areas significantly correlated with each histologic stage of fibrosis (p‐values < 0.05, Fig
1). Lobular MCC showed a statistically significant correlation between stage 0 and stage 1–2 (p‐value 0.0043) but did not correlate with fibrosis in later stages (Fig.
1). Though a positive trend is seen in MCC compared to the stage of fibrosis in both portal tracts and lobular areas, a stronger association is apparent in the portal tracts (Fig
2).ConclusionIn medically complex patients from a large cancer center with exposure to various chemotherapy types and checkpoint inhibitor therapy, mast cell concentrations within portal areas directly correlate with the stage of hepatic fibrosis. This finding further supports the role of mast cells in the development of hepatic fibrosis.Support or Funding InformationMD Anderson Department Chair Research Fund for FellowsFigure 1Figure 2
Quantitative Sudies of Spontaneous Electrical Activities in Detrusor Smooth Muscle Cells towards Understanding Urinary Bladder Overactivities
Authors: Mahapatra Chitaranjan , Manchanda Rohit 
Published: April 2020

ContextThe urinary incontinence (UI) is defined as the involuntary loss of urine and associated with the enhanced spontaneous contractions of the detrusor smooth muscle (DSM). The spontaneously evoked action potentials (sAPs) in DSM cells initiate and modulate these contractions. The DSM is strongly innervated, connecting approximately 16000 afferent and efferent axons from ganglion neurons. It generates sAPs due to the stochastic nature of purinergic neurotransmitter release from the parasympathetic nerve.ObjectivesThe aim of this current study is to understand the putative relationship between the fluctuating ion channel conductances and stochastically release of ATP in generating sAPs.MethodsThe neurotransmitter current was considered as an independent excitatory conductance in the model where gex(t) and Eex are the one‐variable stochastic process conductance and the reversal potential respectively. In addition, Dex and λ1(t) are known as the diffusion coefficients and Gaussian white noise. The point‐conductance is incorporated into a single DSM cell model based on single cylindrical compartment.ResultsThe elicited AP consists an after depolarization and after hyperpolarization phase. The AP peak amplitude and duration are about 15 mV and 40 ms respectively. The stochastic nature of purinergic neurotransmitter is designed by a point process model. Then, the random injection of point process model is conducted to elicit a series of sAPs and membrane depolarization for 5 seconds. The membrane resting potential is held at − 50 mV with a 3 mV of fluctuation. The stochastically depolarization up to 20 mV activates the T‐type Ca2+ channel first and then the L‐type Ca2+ channel to generate action potential. The Figure
1 A illustrates series of AP generation in the DSM cell due to stochastically depolarizations. The model does not evoke any AP and depolarization, when the conductance of T‐type Ca2+ channel is set to zero (Figure
1 B).ConclusionsFrom the results, it is found that the T‐type Ca2+ channel blocker can be used as a new pharmacological target for the UI. In addition, an extended multidimensional models will a provide windows of insight into the factors that govern excitability and contraction in both normal and unstable bladder.Support or Funding InformationThis work is supported in part by Department of Biotechnology (DBT), India (grant number BT/PR12973/MED/122/47/2016).Effects of T‐type Ca2+ channel blockers on AP generation.Figure 1
Understanding the Structure and Function of Type I Acyl Carrier Protein and its Interaction with Growing Acyl Chain
Authors: Garima  , Sundd Monica 
Published: April 2020

Fatty acids are central to the formation of biological membranes and play important roles in metabolism. There are two major classes of fatty acid synthases (FAS); Type I present in mammals and yeast that utilizes large multifunctional enzyme complexes, while type II, harbored by prokaryotes and plants, utilizing discrete monofunctional enzymes. Acyl Carrier Protein (ACP) is a universal and highly conserved player in both the pathways. ACP is modified from apo to holo form by enzyme 4′‐Phosphopantetheinyl Transferase (PPT). PPT enzyme is crucial for growth of many pathogens and is therefore regarded as a potential drug target. Structural and biochemical studies on type II ACP highlight various aspects of ACP‐PPT and ACP‐acyl chain interaction. However, acyl chain binding and dynamic interactions of type I ACP are not well understood. We therefore intend to understand the function of type I ACP, their interaction with bound acyl chain and cognate and non cognate transferases. Literature highlights the importance of type I FAS in the survival and pathogenicity of fungal pathogens like C. albicans and C. parasilopsis. For our studies, we employed type I ACP from Saccharomyces cereviceae and its putative PPT enzyme. Using NMR spectroscopy we show that the growing acyl chain was sequestered up to a certain carbon chain length in the hydrophobic core of the protein employing a unique motif. In conclusion, our studies provide some interesting insights into the structure and function of type I ACP and the molecular mechanism it employs to protect the growing acyl chain. Our findings can provide new avenues for structure based drug designing.Support or Funding InformationDepartment of Biotechnology, India
Simultaneous Non‐invasive Recording of Skin Sympathetic Nerve Activity
Authors: Kusayama Takashi , Wong Johnson , Liu Xiao , He Wenbo , Doytchinova Anisiia , Adams David E. , Chen Lan S. , Everett Thomas H. , Chen Peng-Sheng 
Published: April 2020

Our research laboratory recently invented a new method (neuECG) to simultaneously and non‐invasively record electrocardiogram (ECG) and skin sympathetic nerve activity (SKNA) from humans using standard ECG patch electrodes. These methods are non‐invasive and stable over long periods of time. However, because the neuECG analyses required custom written software, these methods have not been widely used or tested by the scientific community. The purpose of the present study was to develop a method for neuECG recording and analyses using commercially available hardware and software. We used ADInstruments PowerLab to record bipolar electrical signals from the skin of thorax and arms. Wide bandwidth and high sampling rate (10,000/s) were used for recording. The data were then analyzed offline using LabChart Pro 8 software. The digitized neuECG signals are amplified and band‐pass filtered between 0.05 Hz and 150 Hz to display ECG and between 500 Hz and 1,000 Hz to show the SKNA (μV). We full‐wave rectify then integrate all digitized SKNA signals using leaky integrator with 100 ms time constant and display the results over time to simulate the display methods of microneurography (iSKNA, μV.s). For quantitative analyses, we divide the absolute sum of the voltage samples by the number of digitized samples in the chosen window to obtain the aSKNA (μV). We can also examine the distribution of the aSKNA and determine a threshold to separate baseline activity from the burst activity. As compared with the microneurography studies, the neuECG does not require invasive procedures to impale electrodes in nerves and is voltage calibrated. The latter characteristics allow investigators to compare aSKNA, iSKNA and burst duration between subjects or groups. Figure shows the actual neuECG recording. SKNA band‐pass filtered from the digitized signal was converted to iSKNA and aSKNA. Down arrows show premature ventricular contractions. The dotted line indicates the commencement of cold water pressor test that elevated the aSKNA (red double headed arrows) and heart rate. We conclude that neuECG can be recorded and analyzed using commercially‐available hardware and software. These new methods may facilitate the testing and use of this novel technology in experimental biology research.Support or Funding InformationThis study was supported in part by NIH Grants 1OT2OD028190, R01 HL139829, R56 HL71140, TR002208‐01 (Dr. Chen), R42DA043391 (Dr Everett), a Charles Fisch Cardiovascular Research Award endowed by Dr Suzanne B. Knoebel of the Krannert Institute of Cardiology (Drs. Kusayama and Everett), a Medtronic‐Zipes Endowment, and the Indiana University Health‐Indiana University School of Medicine Strategic Research Initiative (Dr. Chen).Example of actual neuECG recordingFigure 1
Small Molecule TREX1 Inhibitors as Novel Immunotherapies
Authors: Hemphill Wayne , Harvey Scott , Gumpena Rajesh , Simpson Sean , Grayson Jason , Smalley Terrence , Salsbury Fred , Hollis Tom , Perrino Fred 
Published: April 2020

Mutations in the 3′ ‐> 5′ exonuclease TREX1 are associated with a spectrum of autoimmune phenotypes in humans and mice. In those affected, pathology is driven by persisting DNA which would normally be degraded by functional TREX1 enzyme. This undegraded DNA stimulates the nucleic acid sensing pathway, cGAS‐ STING, whose subsequent signaling generates a type‐I interferon response. This type‐I interferon response ultimately results in the chronic immune system activation typical of those affected. TREX1 and the cGAS‐ STING pathway have also been implicated in the tumor microenvironment, where TREX1 is believed to degrade tumor‐derived DNA destined to trigger cGAS‐STING signaling. If this tumor‐derived DNA is not degraded, the stimulated cGAS‐STING pathway promotes IFN‐dependent antitumor immunity. Consequently, small molecule stimulators of the cGAS‐STING pathway have shown promise as immunotherapies in animal models. Similarly, mice devoid of TREX1 exonuclease activity exhibit T‐cell dependent resistance to tumor growth. Collectively, these findings indicate that stimulation of the cGAS‐STING pathway activates the immune system, and that TREX1 inactivation represents a novel strategy for this purpose. Furthermore, they demonstrate the immunotherapy potential of such approaches. Thus, we hypothesize that small molecule inhibitors of TREX1 will activate the cGAS‐STING pathway. Studies described here identify small molecule inhibitors of TREX1, determine their mechanism of inhibition, and directly test the hypothesis that they can activate the cGAS‐STING pathway in murine cells. Findings from these studies will generate new structural insights into how TREX1 exonuclease activity may be affected, and have already identified several candidate chemotypes for development into novel immunotherapies.Support or Funding InformationThis work was supported by the NIH, NIAID, NIGMS, Lupus Research Alliance, Wake Forest Innovations, and a Sandy Lee Cowgill Memorial Scholarship.D18N mice display T‐cell dependent tumor resistance.5×106 H31ml tumor cells were injected subcutaneously into WT and D18N mice, and tumor volume and survival tracked daily. Data in ‘B’ are separate study from ‘A’. For each study, eight to 16 mice were examined in two to four independent experiments. (A) D18N mice have decreased tumor growth in a mouse tumor model. (B) D18N model tumor‐resistance is T‐cell dependent. WT and D18N were treated with αCD8, αCD4, or isotype‐control antibodies. Arrows indicate D18N mice without treatment, or with CD8+ / CD4+ T‐cell depletion.Figure 1
Preliminary study of in‐situ 3D bioprinted nano‐silicate biopolymer scaffolds for muscle repair in VML defects
Authors: Ahuja Neelam , Awad Kamal , Fiedler Matthew , Aswath Pranesh , Brotto Marco , Varanasi Venu 
Published: April 2020

IntroductionThe traumatic or surgical loss of skeletal muscle with resultant functional impairment (volumetric muscle loss (VML)) is a challenging clinical problem. The current standard treatment care for VML is autologous tissue grafts and physical therapy, which still faces the challenge of donor site morbidity in large defect cases. 3D printing techniques for tissue engineering allows the precise control of micropatterning by determining the dimensions of filaments, the size and shape of pores and the percentage of porosity of the scaffold. Our hypothesis is using 3D printing of biopolymer‐nanosilicate scaffolds in‐situ will enhance muscle regeneration in VML defects.Materials and MethodsMethacrylated gelatin (MAG) biopolymer Laponite (Lp) nano‐silicate scaffolds were prepared as a bio‐ink using a solution‐precipitation process. Our study includes designing and optimization of 3D printing parameters for printing MAG‐Lp scaffolds and in‐vitro evaluation of cell viability (live‐dead), proliferation, and differentiation of mouse myoblast precursor cells on these scaffolds. We further analyzed the in‐situ (directly in the defect) 3D printing (using a robocaster) in a rat VML model in tibialis anterior muscle to evaluate the feasibility, precision, flexibility and attachment of the scaffold to the defect.ResultsSilicon ion enhanced cell proliferation and differentiation (fusion index by 1.5 times and overexpression of myogenic factors) in mouse myoblast precursor cells as compared to control. Qualitative live/dead analysis showed more abundance of live myoblast precursor cells (green) and decreased number of dead cells (red) on MAG‐Lp surface as compared to MAG (Fig.
1). We also show successful printing abilities of MAG‐Lp scaffolds in the volumetric muscle loss defect (Fig.
2) along with increased attachment to the adjacent tissues. The animal also showed ability to stand on/flex the surgical limb, with sufficient grip strength to perform normal activity, immediately after recovering from the surgery.ConclusionOur preliminary findings conclude that 3D printing of MAG‐Lp can enhance myogenesis in myoblasts. We further conclude that successful proof of concept of an in‐situ bioprinting method of reconstructive scaffolds can be used for immediate repair and loading of muscles for VML injury treatment.Support or Funding InformationNIH/NIDCR (R03DE023872‐01, 1R56DE027964‐01A1‐01), Texas STARs award, Departmental Start‐up Funds, University of Texas at Arlington, College of Nursing and Health Innovation, Arlington, TXshowing live/dead assay on MAG and MAG‐Lp scaffolds. Green represents live cells, whereas red represents dead cells.Figure 1showing In‐situ 3D printing biopolymer scaffold in a rat volumetric muscle loss model in tibialis anterior muscle (20–25% muscle loss) along with UV/photo curingFigure 2
Histopathological Impact Of Nematodes On The Gastric Mucosa Of The American Alligator (Alligator Mississippiensis)
Authors: Yesudas Rekha , Young Bruce 
Published: April 2020

Nematodes are adapted to inhabit a vast array of niches. They have evolved tremendous strategies to survive and succeed in a variety of hosts and host tissues. Intestines represent an ideal habitat for a large number of parasites, compared to the stomach. In the present study we examined the stomach of 14 sub‐adult, wild‐caught American alligators (Alligator mississippiensis) from the coastal region of Louisiana. Nematodes were found in four of the alligators (~29%). Two different parasites were found; Dujardiascaris waltoni (Nematoda: Heterocheilidae) and Ortleppascaris sp.? (Nematoda: Ascaridoidea). D. waltoni was found loose among the stomach contents, whereas Ortleppascaris sp.? was found embedded in the stomach wall. Ortleppascaris was associated with multifocal lesions in the gastric mucosa. These lesions were roughly 4 mm in diameter and housed multiple parasites. The lesions caused a clearly visible penumbra of mucosal damage. As the parasites invaded the gastric mucosa, they induced a granulomatous inflammation of the mucosa and submucosa. There was a marked eosinophilic necrosis, a cellular response that produces the penumbra around the invasive nodules. The impacted worms burrowed into the mucosa, and were surrounded by an eosinophilic exudation, resulting in an elevated nodule of worms and mucosa. This presumably represent proliferating host mucosal tissue responding to the parasitic challenge. Alligators have “typical” vertebrate gastric pits and glands in the stomach mucosa. Mucosal burrows of the nematodes led to a loss of the gastric pits in the infected zone and digestive efficiency of the host. Previous studies have described keratitis and enteritis associated with nematode infections in crocodilians, but similar reports are lacking from Alligator mississippiensis. Alligators and crocodiles have remarkably effective immune systems, which enable them to fight against microbial infections and allow quick wound healing. We hope to expand our understanding of host‐parasite relations in this important species.Host response/eosinophil exudation in infected gastric mucosaFigure 1Nematodes burrowed in to the stomach mucosaFigure 2
Extract of Foeniculum vulgare seed effect on UV‐induced photoaging‐associated ion channel and collagenase
Authors: Woo Joohan , Kim Yung Kyu , Kim Woo Kyung 
Published: April 2020

Skin aging is mediated by genetics, hormonal changes, metabolic processes, and environmental stress. UV radiation is concerned as a major risk factor for skin aging, including wrinkles, and melanoma. Studies suggested that calcium ion channels, such as TRPV1 and ORAI1, are related with melanogenesis, skin wrinkling, and inflammation. Foeniculum vulgare has been used to treat inflammatory diseases. Recent studies showed that it protects against UV radiation‐induced skin photoaging. However, its effect on the ion channels related to UV‐induced photoaging is unclear. Therefore, this study aimed to evaluate the antagonistic effects of F. vulgare extract, fractions, and constituents on TRPV1.Electrophysiological analysis results showed that, at 100 μM, F. vulgare extract and fractions potently inhibited capsaicin‐induced TRPV1 [methanol extract (91%), n‐hexane (94%), and methylene chloride (100%) at −60 mV]. The methanol extract of F. vulgare inhibited MMP‐1 (collagenase) production (35% at 3.75 μg/mL), compared with control. These results suggested that the compounds in F. vulgare extract and fractions possessed anti‐aging effects that can decrease wrinkles and maintain elasticity by disrupting MMP‐1 production activated by the MAPK pathway. Unfortunately, compounds isolated from the hexane and methylene chloride fractions could not suppress ITRPV1. Surprisingly, even though capsaicin increased ITRPV1, the γ‐terpinene, limonene, scopoletin, and bergapten isolated from these two fractions slightly potentiated ITRPV1. Although 4 compounds of F. vulgare exerted an agonistic effect on TRPV1, the compounds with antagonistic effect have not been discovered. Our study suggested that F. vulgare had potential therapeutic effects on UV‐induced photoaging for therapeutic and cosmetic applications.Effect of F. vulgare extract and fractions on human TRPV1 (hTRPV1) current under capsaicin (CAP) treatmentFigure 1Effects of F. vulgare extract and fractions on UV‐induced melanogenesisFigure 2
Effects of Short‐term Warm Water Bathing on Cardiac Baroreflex Sensitivity in Healthy Men
Authors: Sugawara Jun , Tomoto Tsubasa 
Published: April 2020

Bath‐related fatal (sudden cardiac arrest) and non‐fatal accident (dizziness and disturbance of consciousness) have been reported frequently in Japan. Cardiac baroreflex sensitivity (BRS) may relate to these symptoms. However, underlying mechanism(s) is poorly understood. With this information as background, we determined the effects of short‐term warm water immersion on cardiac BRS in healthy men. Twelve apparent healthy men underwent the single sit‐stand test (e.g., 5‐min sitting on the floor of bathtub and then standing up and remained standing for 5 min quietly) twice (e.g., without and with 41 °C warm water at the heart level). Beat‐to‐beat blood pressure was continuously recorded through the protocol (via Finometer). BRS was evaluated by the sequence method from heart rate response to the standing‐induced abrupt drop of mean arterial pressure (MAP). Additionally, after the each test, brachial‐ankle pulse wave velocity (baPWV: an index of arterial stiffness) was measured in the supine position. During the bathing, heart rate and cardiac output (estimated by Modelflow method) were significantly increased but MAP, stroke volume, total peripheral resistance, and tympanic temperature did not change significantly (vs. pre‐bathing). Regarding the single sit‐stand test, there were no significant changes in the baseline (sitting) MAP (from 73±16 to 77±16 mmHg, P=0.322) and the standing‐induced decreases in MAP (from −33±12 to −33±13 mmHg, P=0.970) between pre‐ and post‐bathing, but MAP‐HR slope was blunted after the bathing (from −0.47±0.26 to −0.35±0.25 bpm/mmHg, P=0.024, Figure
1). The baPWV was significantly decreased after the bathing (vs. pre‐bathing (from 12.2±1.2 to 11.8±1.2 m/sec, P=0.002). Change in MAP‐HR slope correlated with corresponding change in baPWV (r=0.610, P=0.036, Figure
2). Our results indicate that: (1) even a short‐term warm water bathing deteriorates cardiac BRS although it did not induce greater orthostatic hypotension, and (2) the change in cardiac BRS with the short‐term bathing is associated with the change in arterial stiffness. Further study to determine the impact of prolonged warm water bathing is needed.Support or Funding InformationThis study was supported by Japan Health & Research Institute Research Grant.Cardiac barorefex sensitivity (BRS) gain and brachial‐ankle pulse wave velocity (baPWV) before and after 5‐min warm‐water bathing. Data are mean and SD. Thin lines indicate individual data.Figure 1Relationship between 5‐min bathing induced changes in baPWV and cardiac BRS gain.Figure 2