bluuebunny/crossref_metadata_2025_split huggingface dataset:
Magnesium ion disrupts LAP surface re‐association of Listeria monocytogenes by dissociation of InlB
Authors: Liu Dongqi , Bai Xingjian , Tenguria Shivendra , Bailey Talyor , Drolia Rishi , Singh Atul K. , Amalaradjou Mary Anne Roshni , Seleem Mohamed , Bhunia Arun K. 
Published: April 2020

Listeria monocytogenes crosses intestinal epithelial barrier for the systemic spread, which is facilitated in part, by Listeria adhesion protein (LAP). LAP, a housekeeping enzyme called acetaldehyde alcohol dehydrogenase, is present in both pathogenic and nonpathogenic Listeria. LAP is secreted with the aid of SecA2 translocon, and its’ re‐association on the surface of pathogenic Listeria i.e., L. monocytogenes is critical for its function. However, the cell wall component(s) involved in LAP interaction is unknown, which was investigated.Using L. monocytogenes F4244 (WT), an isogenic lap− strain and purified recombinant LAP in the Caco‐2 cell adhesion assay and Western blot, we examined LAP re‐association on lap− cells. Extraneous added LAP restored adhesion capacity of lap−, but not of L. innocua to Caco‐2 cells, suggesting the presence of LAP‐interacting receptor(s) only on pathogenic bacteria.To locate LAP interaction partners, co‐immunoprecipitation of WT cell wall fraction using anti‐LAP mAb was analyzed by SDS‐PAGE. Selected protein bands were subjected to MALDI‐TOF protein identification showing involvement of internalin B (InlB).Next, we investigated the effect of MgCl2 pretreatment on LAP re‐association on the cell surface, cell‐free supernatant, and consequent bacterial adhesion. Mg2+ is a wall teichoic acid (WTA) charge neutralizing agent that reduced levels of InlB binding on the WT cell wall, resulting in surface disassociation of LAP. However, only partial surface reduction of LAP was observed in ΔinlB strain suggesting more than one ligand is involved.In conclusion, our results indicate that both InlB and WTA may participate in LAP re‐association on L. monocytogenes cell surface, which is essential for its pathogenic activities.Support or Funding InformationThe technical assistance of Uma Aryal, Valerie Ryan, and Aaron Gall and funding from USDA‐ARS project number 59‐8072‐6‐001 through the Center for Food Safety Engineering at Purdue University are acknowledged.LAP Surface Ligand Fishing ExpeditionAs the experiment scheme shown (A), several Co‐immunoprecipated candidates were visualized by Sliver staining. Red arrow point to a potential receptor around 70 kDa (B). Abundance of each identified protein was graphed according to relative protein abundance (C). Internalin B is the first protein present only in pathogenic strain.Figure 1MgCl2 Treatment Affects LAP Dissociation and Bacterial AdhesionWB protein analysis in Supernatant and Cell Wall Fraction (A), growth curve (B) and adhesion (C) of LmWT grow with additional MgCl2 in TSBYE.Figure 2
High expression of Nucleobindin‐2 is associated with poor prognosis in gastric cancer
Authors: Okada Junichi , Okada Shuichi , Yamada Masanobu 
Published: April 2020

Nucleobindin‐2 (NUCB2) is a 396‐amino acid protein, cleaved into the N‐terminal nesfatin‐11–82, nesfatin‐285–163 and the C‐terminal nesfatin‐3166–396. NUCB2 contains a signal peptide, a leucine zipper structure, two Ca2+ binding EF‐hand domains, and has a wide variety of basic cellular functions. NUCB2 is also a precursor protein of nesfatin‐1, which was originally identified in hypothalamic nuclei, and which is a regulatory factor involved in the central control of food intake and energy balance.There are several reports indicating that NUCB2 is also expressed in various human peripheral tissues. Moreover, recent studies have reported that high levels of NUCB2 mRNA and protein are a potent prognostic factor for prostate cancer, endometrial carcinoma, and breast cancer. However, the clinicopathological significance of NUCB2 expression in gastric cancer has still not been elucidated. Therefore, we examined NUCB2 expression in a large number of gastric cancer patients, using immunohistochemistry, to explore its clinicopathological significance. To explore this, we aimed to investigate the NUCB2 expression in gastric cancer tissues and adjacent non‐tumor tissues and its potential relevance to clinicopathological factors and prognosis using immunohistochemistry analysis. In our study, NUCB2 level in gastric cancer tissues was higher than in non‐tumor tissues. A high expression of NUCB2 is significantly associated with tumor depth, lymph node metastasis, lymphatic invasion, venous invasion and clinical stage. Furthermore, the expression level of NUCB2 protein was independent predictor of progression‐free survival.In summary, NUCB2 might play a crucial role in gastric cancer development and could serve as an independent predictor of prognosis of gastric cancer patients.Progression‐free survival and overall survival curves of 328 gastric cancer patients divided into two groups according to NUCB2 expression levels. The significant differences were given as p = 0.0214 for progression‐free survival and p=0.0211 for overall survival.Figure 1
Effects of Obesity and Sleep Apnea on Sympathetic Neural Activity during Early Pregnancy in Humans: A Pilot Study
Authors: Hsieh Yi-Jin , Yoo Jeung-Ki , Parker Rosemary S. , Clark Amanda M. , Takeda Ryosuke , Fu Qi 
Published: April 2020

Maternal obesity is a major risk factor for adverse pregnancy outcomes and this increased risk is attributed, at least in part, to obstructive sleep apnea (OSA; defined as an Apnea and Hypopnea Index ≥5). However, whether obesity and OSA increase the risk of adverse outcomes through sympathetic neural mechanisms is unknown. We hypothesized that obese women with OSA would have augmented sympathetic nerve activity during pregnancy.Twenty‐five women within 10 weeks of gestation were studied; 13 of them were normal‐weight (BMI 18.5–24.9 kg/m2) while 12 were obese (BMI ≥30 kg/m2). Home‐based sleep testing was performed using a portable device (WatchPat) and 24‐hour ambulatory blood pressure (BP) was monitored in all subjects. Muscle sympathetic nerve activity (MSNA), beta‐by‐beat BP, and heart rate were continuously recorded during supine rest in our laboratory.We found that obese pregnant women had higher 24‐hour systolic (mean±SD; 129±13 vs 108±5 mmHg, P<0.001) and diastolic (72±9 vs 62±4 mmHg, P=0.0013) BP compared with normal‐weight pregnant women. Resting MSNA trended to be greater in obese than normal‐weight women (burst frequency: 31±20 vs 19±7 bursts/min, P=0.063; burst incidence: 40±26 vs 26±10 bursts/100 heartbeats, P=0.108; total activity: 358±283 vs 189±95 a.u./min, P=0.075). Preliminary data showed that among obese pregnant women, those with OSA (n=3) had much higher systolic (140±8 vs 125±12 mmHg)and diastolic (82±3 vs 69±8 mmHg) BP compared to those without OSA. In addition, MSNA appeared to be greater in obese women with OSA versus those without OSA (57±12 vs 20±10 bursts/min; 72±21 vs 26±11 bursts/100 heartbeats).These preliminary results indicate that obesity and OSA may be associated with augmented sympathetic activity and elevated blood pressure early on during pregnancy, leading to the development of adverse maternal‐fetal outcomes.Support or Funding InformationSupported by the NIH R01 grant (HL142605).24 hour ambulatory blood pressure was greater in obese pregnant women with sleep apnea than pregnant women with obesity only.Figure 1Impact of sleep apnea and maternal obesity on resting MSNA during early pregnancy.Figure 2
Sildenafil reverses the right ventricular hypertrophy induced by hypoxia but does not bring down the expression of MHC‐β to normal value
Authors: Al-Janabi Mukhallad M. , Khatib Said Y. 
Published: April 2020

IntroductionMyosin is the major player in structure and function of cardiac muscle. There are two sub‐types of myosin heavy chain in cardiac muscle; myosin heavy chain‐α (MHC‐α) and myosin heavy chain‐β (MHC‐β). The contractile properties of the cardiac muscle are determined largely by types and the expression of the myosin heavy chain (MHC) which is altered by many physiological and pathological conditions.ObjectivesIn this study we investigated the effect of hypoxia and sildenafil treatment on myosin heavy chain isoform in hy poxia ‐ induced hypertrophied right ventricle.MethodsRight ventricular hypertrophy was induced in 45 mice by exposing them to hypoxic stimulus (11% ambient oxygen) in normobaric chamber for 20 days. The animals were distributed randomly into three groups: first group served as control (CO), second group exposed to hypoxia for 20 days without sildenafil treatment (HY) and third group was given sildenafil orally at dose of 30 mg.Kg−1.day−1 plus exposure to hypoxia for 20 days (HS). MHC isoforms were calculated using two ELISA kits containing antibodies against α and β MHC and by SDS‐PAGE (Mybiosource‐USA). The HY group in comparison with CO group, showed a significant increase in right ventricle weight/left ventricle plus septum ratio (Fulton’s ratio). HS group showed significant decrease in Fulton’s ratio compared with HY but not with CO. Expression of MHC‐β isoform was significantly increased in HY group compared with CO. There was no significant difference in MHC‐β between HY group and HS group. Expression of more beta myosin heavy in hypoxic group is beneficial to the heart since hearts with more MHC‐β have more economic contraction.Conclusionsildenafil reversed the right ventricular hypertrophy induced by hypoxia but did not bring down the expression of MHC‐β to normal value.Support or Funding InformationDeanship of research at Jordan University of Science & TechnologyEffects of hypoxia and hypoxia with sildenafil on myosin heavy chain isoforms.Expression of cardiac myosin heavy chains in different groups: Hypoxia caused expression of more MHC‐β and sildenafil failed in revering the changes caused by hypoxia. Control (CO), hypoxia for 20 days without treatment (HY) and hypoxia for 20 days plus treatment with sildenafil (HS). *** P < 0.01 HY and HS vs COFigure 1
Improved Difluoroboron β‐Diketonate Poly(lactic acid) Nanoparticles for Monitoring Wound Oxygenation
Authors: Bruce Anthony C. , Zhuang Meng , DeRosa Christopher A. , Corliss Bruce A. , Fraser Cassandra L. , Peirce Shayn M. 
Published: April 2020

We have previously demonstrated the ability of a difluoroboron β‐diketonate poly(lactic acid) material, BF2nbm(I)PLA, hereafter nbm, to detect tissue oxygen levels in murine cutaneous wound beds
1. This relies on oxygen‐insensitive fluorescence (F) as an internal standard and oxygen‐sensitive phosphorescence (P) as a sensor. In an effort to improve imaging agent clearance and ease of manufacture, as well as to optimize emission wavelengths to color channels of RGB cameras, a new dithienyl dye, BF2dtm(I)C12H25‐, hereafter dtm, with red phosphorescence was developed. The dtm dye was mixed with a blue fluorescent dye, BF2dbm(F)OCH12H25, hereafter dbm, and coprecipitated with PLA to form oxygen sensing nanoparticles.The objective of this study was to verify the utility of the newly developed dtm/dbm nanoparticles for sensing tissue oxygen levels in murine wound beds. To achieve this, we monitored the relative tissue oxygen levels of healing full‐thickness cutaneous wounds in C57BL/6J mice over the course of one week with both dtm/dbm and nbm using an RGB camera mounted to a microscope and custom MATLAB software.We found the new dtm/dbm nanoparticles yielded nearly identical trends over time in wound bed F/P versus nbm. The dtm/dbm nanoparticles did not impair wound closure and demonstrated less adhesion to healing tissues relative to nbm. Dtm/dbm retains the tissue oxygen‐sensing capability of nbm while having fewer synthetic steps, better phosphorescence alignment with red RGB channel, improved solubility, and less retention in wound tissue than nbm.Support or Funding InformationUVA‐Coulter Translational Research Project #155234Verification of dtm/dbm in murine cutaneous woundsA. Dtm/dbm generated the same trends in wound bed tissue oxygenation over time as nbm. B. Dtm/dbm did not impair wound healing versus nbm or saline control. C. Dtm/dbm displayed reduced particle retention in wounds after multiple applications. Scale bar = 1 mmFigure 1
Electronic Depth Adjustment in an Automatic Chest Compression Device for Rodents – Design and Implementation of a LabVIEW‐based Control Unit
Authors: Riess Jonathan D. , Balzer Claudius , Riess Matthias L. , Cleveland William J. 
Published: April 2020

IntroductionDespite best efforts of cardiopulmonary resuscitation (CPR), many patients still die or suffer severe cardio‐cerebral damage following cardiac arrest (CA). The quality of manual chest compressions (CC) rarely adheres to current guidelines with regards to CC rate, depth and fraction of time spent during CPR. Automated mechanical CC devices offer a reliable improvement in both the clinical arena and in research models. We describe the mechanical and programming aspects of the development of a depth controller for a pneumatic CC device for CPR in rodent CA model.Methods & ResultsIn order to electronically control the amplitude/depth of the vertical CC piston (Weil Institute of Critical Care Medicine, Palm Springs, CA; Fig
1) that is pneumatically displaced between 0 and maximally 2 cm 200 times min−1 (duty cycle 1:1), a horizontally moving wedge was connected to a rack gearbox driven by a low‐voltage motor (Fischertechnik, Waldachtal, Germany) along a fixed rack. The unit was connected to a horizontally sliding potentiometer to monitor the exact position of the wedge and, therefore, the piston displacement which itself was monitored through a second, vertically sliding potentiometer. Both potentiometers were connected to an USB 6343 data acquisition system (DAQ, National Instruments, Austin, TX). The motor was connected to two analogue outputs of the DAQ (for movement and direction) via a commercially available low‐voltage amplifier. Stepwise movement of the wedge in either direction could be controlled by a momentary rocker switch connected to two analogue input channels of the DAQ (for deeper and shallower compressions) and its LabVIEW software which was programmed to allow precise control of the wedge’s position.ConclusionsWe report the design and construction of an inexpensive, LabVIEW‐based control unit to electronically adjust CC depth in a rodent model of CA and CPR. Neither the hardware, nor the software required to perform this highly specialized application were commercially available. This custom‐designed and ‐built device achieved its purpose of allowing electronic control and recording of the CC depth in this rodent model with high reliability and precision. Moreover, it constitutes the necessary basis of pre‐programmed or feedback‐assisted adjustments of variable CC depth in the future to improve neurologically favorable outcome after CA.Support or Funding InformationThis work was supported, in part, by institutional funds, and a Merit Review Award (I01 BX003482) from the U.S. Department of Veterans Affairs Biomedical Laboratory R&D Service.Pneumatic chest compression device for rodents with added control unit for electronic depth adjustment.Figure 1
Development of picomolar liver receptor homolog‐1 agonist
Authors: Cato Michael Lee , Cornelison Jeffery , Flynn Autumn , D’Agostino Emma , Jui Nathan , Ortlund Eric 
Published: April 2020

Nuclear receptors are ligand‐regulated transcription factors that alter target gene expression in response to the binding of lipophilic signaling molecules. Liver receptor homolog‐1 (LRH‐1) is a nuclear receptor that has shown promise as an anti‐diabetic therapeutic in murine studies and regulates bile acid biosynthesis, steroidogenesis, and glucose homeostasis, making it an attractive target for treating a variety of metabolic diseases. Although phospholipids are the putative endogenous ligands for LRH‐1, they are insoluble, readily remodeled and hydrolyzed, and are only efficacious at very high concentrations, making development of synthetic agonists critical for targeting LRH‐1 in clinical and laboratory contexts. Using structural data that delineated the binding pose of the only class of synthetic agonists known for LRH‐1, we have conducted structure‐activity relationship studies to enhance potency, efficacy, and solubility of small molecules. Specifically, we targeted residues deep within the binding pocket that radically enhance potency, which has led to the first low‐nanomolar LRH‐1 agonist (“6N”). We have also targeted residues contacted by the headgroups of activating phospholipids as a complementary strategy for activation. These modifications resulted in our most efficacious small molecule (“10CA”). To capture the characteristics of both compounds (6N and 10CA), we have synthesized a chimeric small molecule that includes the key moieties of 6N and 10CA responsible for high affinity and efficacy, respectively. Excitingly, this new hybrid compound binds with picomolar affinity, is more efficacious than 10CA, and elicits LRH‐1 target gene expression to a greater degree than either 6N or 10CA alone. The hybrid molecule is our most effective agonist to date, providing a critical tool for both studying LRH‐1 biology and treating a range of metabolic disorders.Support or Funding InformationNIH‐5T32GM008367‐29 (BCDB Training Grant) | NIH‐5R01DK115213‐02 | Emory Catalyst ProgramBinding mode and residue contacts of leading agonists.Figure 1Binding and in‐cell reporter data demonstrating affinity, potency, and efficacy of the hybrid compound relative to our two leading agonists.Figure 2
Exercise‐mediated Wall Shear Stress Attenuates NLRP3 Inflammasome‐induced Endothelial Dysfunction in Obesity
Authors: Lee Jonghae , Hong Junyoung , Umetani Michihisa , Juttu Aishwarya , Park Yoonjung 
Published: April 2020

PURPOSENLRP3 inflammasome plays a pivotal role in obesity‐induced vascular pathology through caspase‐1‐mediated IL‐1β processing, while precise mechanism by which exercise ameliorates the inflammasome remains unknown. Thus, we investigated the effect of exercise on endothelial dysfunction in obesity via NLRP3 inflammasome‐dependent mechanisms.METHODSC57BL/6J male mice were provided a control low‐fat diet (LFD; 4.5% calories from fat) or a high‐fat diet (HFD; 45% calories from fat) with a running wheel access (LF‐RUN or HF‐RUN) or a sedentary no‐wheel condition (LF‐SED or HF‐SED) for 12 to 14 weeks. Endothelium‐dependent vasodilation in response to increasing acetylcholine (ACh, 1 nmol/L to 10 μmol/L) was assessed using intact mesentery arteries isolated from mice. Mouse microvascular endothelial cell (MVEC) line EOMA was treated with palmitate (PA, 20 μmol/L) and laminar shear stress (LSS, 20 dynes/cm2): Static control (STT), PA, LSS, and LSS+PA. The NLRP3 inflammasome axis, including NLRP3, ASC, caspase‐1, p20, and IL‐1β in mice aorta and MVECs was examined by RT‐qPCR, FLICA and DAB staining and western blot.RESULTSHFD significantly increased obesity‐associated parameters while these were attenuated by running exercise. Obesity greatly decreased ACh‐induced endothelium‐dependent vasodilation compared to LF‐SED. Exercise improved reduced vasodilator response to ACh in obese mice. Incubation of caspase‐1 inhibitor (10 μmol/L) to the isolated vessel restored obesity‐induced impairment of vasodilator response similar to one of LF‐SED, LF‐RUN, and HF‐RUN. Exercise‐induced decrease of caspase‐1 and IL‐1β in endothelial cells of obese mice aorta was determined by immunohistochemistry using FLICA and DAB, respectively. Likewise, in in vitro experiment, LSS diminished inflammasome components in PA‐treated MVECs.CONCLUSIONThese findings suggest that exercise attenuates against obesity‐induced endothelial dysfunction through a shear stress‐dependent mechanism. Our finding suggests that exercise may be a therapeutic strategy for inflammation‐related vascular dysfunction in obesity.Exercise attenuates IL‐1β in endothelium of obese mice aorta.Immunohistochemical analysis of IL‐1β expression (dark brown precipitate) in the endothelial layer of the aorta. Summarized data showing area percentage of the endothelium positive for IL‐1β in aorta (n=4–6). Values are means ± SEM. * P<.05 vs. LFSED; # P<.05 vs. HF‐SED.Figure 1Inhibition of caspase‐1 abolishes endothelial dysfunction in mesentery arteries of obese mice.Incubation of the caspase‐1 inhibitor, Ac‐YVAD‐cmk, in isolated mesentery arteries from obese mice showing improved ACh‐induced endothelium‐dependent vasodilation in HF‐SED + Ac‐YVAD‐cmk (n = 3) similar to the levels of LF‐SED (n=8), HF‐RUN (n = 9) and HF‐RUN + Ac‐YVAD‐cmk (n = 7). Values are means ± SEM. ## P<.01 vs. HF‐SED.Figure 2
Discovery of novel compound promotes neurogenesis by activation of mTOR signaling
Authors: Ashraf-Uz-Zaman Md , Vaidya Bhuvaneshwar , Abbruscato Thomas , German Nadezhda 
Published: April 2020

Neurodegenerative disorders are continually increasing in incidence due to higher life expectancy. Currently, limited therapeutic strategies are available for affected patients, most targeting symptomatic treatment. mTOR pathway is identified as important player in neurogenesis that modulates healthy neuron growth and differentiation. Moreover, many neurodegenerative disorders are associated with downregulated mTOR signaling. However, mTOR activation can also lead to uncontrolled cell proliferation, promoting cancer. This careful balance limits the development of mTOR as a target for modulation of neuronal growth. Recently, selected classes of antidepressants were reported to activate mTOR pathways, causing dendritic outgrowth and increased synaptic protein levels. Although results were promising, they require treatment with up to 100 μM of tested compounds.To identify potential utilization of penfluridol, an anti‐psychotic agent, as a neuron growth modulator, we have screened this compound and its analogs using reported methodology. Although no activity was observed for PFL, one of its analogs, PF331, promoted neurogenesis in primary neurons. In particular, treatment with PF331 (100 nM,1 μM) for 5 days during reoxygenation (post‐stroke conditions) resulted in improved neurite outgrowth. At the same time, no effect on the viability of the primary neurons was observed under normoxic conditions and during the reperfusion stage. Further, we have established that observed activity was not related to the overstimulation of mTOR pathway, but rather to the compound’s ability to restore mTOR activation to the pre‐stress levels. We hypothesized that PF331 will have minimal impact on cancer proliferation and confirmed it in vivo, where our compound had no effect on cancer growth. Stroke is one of the pathological conditions characterized by hypoxia‐induced damage to neurons. Hence, we have proposed to evaluate our compound for the ability to lessen hypoxia‐induced damage using in‐vivo stroke model with prior confirmation of the ability of PF331 to cross the BBB in vivo and to achieve desired levels in the brain.In conclusion, PF331 has shown the ability to promote neurite outgrowth in primary neurons and restore mTOR activation in the hypoxic in vitro model. Because of its biological activity and favorable in vivo PK profile, we envision that this compound is a potential candidate for the treatment of neurodegenerative diseases, such as stroke.Evaluation of the effect of PF331 on primary neuronsEvaluation of the effect of PF331 on of primary neurons. Neurons were subjected to 3 hrs of OGD followed by 4 (A and B)/ 5 (C) days of oxygenation to mimic ischemia reperfusion injury. (A–B) OGD reperfusion (OGDR) down‐regulates mTOR and ERK compared to normoxic (Nor) condition whereas treatment with Compound PF331 activates mTOR and ERK. (C) PF331 facilitates dendritic outgrowth and increased number of synapses.Figure 1