bluuebunny/crossref_metadata_2025_split huggingface dataset:
G protein‐coupled estrogen receptor (GPER) mediates the inhibitory action on P2Y receptor‐mediated Ca2+ signalling and cytokine secretion in human bronchial epithelia (LB831)
Authors: HAO Yuan , CHOW Wai Ming Alison , YIP Chung Yin Wallace , CHAN Wood Yee Woody , CHENG Hon Ki Christopher , KO Wing Hung 
Published: June 2021

The airway epithelium plays a central role in respiratory physiology through its transport and immunological functions. Our previous study suggested that P2Y purinergic receptors are expressed in airway epithelia and play a significant role in regulating transepithelial ion transport. P2Y receptors are G protein‐coupled receptors classically signal through Gq, resulting in an increase of intracellular Ca2+ concentration and activation of Ca2+‐dependent downstream signaling pathway. In addition, P2Y receptors are also involved in asthmatic inflammation.Estrogen (E2) is an important hormone in human physiology. In addition to the classical nuclear hormone receptors ERα and ERβ, a novel estrogen receptor, G‐protein coupled estrogen receptor (GPER/GPR30), was recently identified and found to be involved in both rapid signaling and transcriptional regulations. The action of GPER is unclear but it has been implicated in mediating anti‐inflammatory responses. Our study aimed to investigate the pro‐inflammatory action of P2Y receptors and the interaction between GPER and P2Y receptor‐mediated signaling pathways.Our results demonstrated the expression and localization of GPER in both primary normal human bronchial epithelial cells (obtained from ScienCell Research Laboratories, San Diego, CA, USA) and 16HBE14o‐ cell line using qPCR, western blotting, and immunofluorescence. Activation of GPER by E2 or its specific agonist, G1, rapidly attenuated a nucleotide‐evoked Ca2+ increase, while the specific GPER antagonist, G15, reversed this GPER‐mediated inhibition. Furthermore, E2 and G1 also inhibited nucleotide‐induced cytokine release. Our results provide the first evidence that human bronchial epithelia express GPER, which interact with the P2Y receptor‐mediated calcium signalling pathway and cytokine secretion. Moreover, the anti‐inflammatory role of GPER may be due to its opposing effect on the pro‐inflammatory pathway activated by the P2Y receptors in inflamed airway epithelia.Grant Funding Source: Supported by Research Grant Council General Research Fund (#2140730) awarded to W. H. Ko.
Unoprostone protects against cell death, reactive oxygen species, and apoptosis in a model retinitis pigmentosa system using expression of wild type and mutant rhodopsin (LB832)
Authors: Cuppoletti John , Chakrabarti Jayati , Malinowska Danuta , Ueno Ryuji 
Published: June 2021

Objective. This study was to determine if unoprostone protects against deleterious effects evident in retinitis pigmentosa (RP). With the P23H mutation in rhodopsin, it is a dominant negative disease, causing the unfolded protein response (UPR) and ER stress. This leads to reactive oxygen species (ROS) production, apoptosis and ultimately cell death.Methods. Flp‐In T‐Rex‐293 cells were transfected with wild type (WT) or P23H mutant (MUT) rhodopsin in pcDNA5/FRT/TO with pOG44 under control of a tetracycline (tet) promoter. Cell growth, ROS and caspase were measured in non, mock, WT and MUT transfected cells with 1 µg/ml tet ± 100 nM unoprostone. ROS was measured with (5&amp;6)‐chloromethyl‐2’,7’‐dichlorodihydrofluorescein diacetate,acetyl ester and caspase was detected using FITC‐VAD‐FMK.Results. There were no significant differences between non‐, mock‐ and WT (WT) cells ± unoprostone. At 120 h (n=3) the number of WT cells increased from 54.6 ± 1.2 to 111.7 ± 3.1, while the number of MUT cells decreased from 56.7 ± 3.6 to 19.0 ± 1.3. Unoprostone increased significantly (P<0.005) the number of MUT cells to 32.0 ± 0.5. ROS (FU, n=6) increased from 332.9 ± 0.7 in WT cells to 2816.8 ± 6.4 in MUT cells and this was significantly decreased (P<0.0005) to 1331.0 ± 3.0 with unoprostone. Caspase (FU, n=6) at 120 h increased from 365.7 ± 5.3 in WT cells to 895.5 ± 10.2 in MUT cells and this was significantly reduced (P<0.0005) to 523.2 ± 11.3 with unoprostone.Conclusions. The data shows that unoprostone can slow the pathological events (ER stress, ROS production, and apoptosis) and cell death seen in RP with UPR. Increased ROS generation and apoptosis are also seen in age related macular degeneration (AMD), suggesting unoprostone should also be investigated in dry AMD.Grant Funding Source: Supported by Sucampo AG
Metastatic tumor cells induce a crosstalk between endothelial p38 and ERK1/2 MAPK pathways activating endothelial MMP system (LB838)
Authors: Glinskii Olga , Glinsky Vladislav 
Published: June 2021

Our recent results demonstrate that metastatic cell adhesive interactions with endothelial cells (EC) are stabilized by the endothelial integrin α3β1 leading to the activation of p38 and ERK1/2 pathways in endothelial cells when they interact with tumor cells. Here we demonstrate that, while FAK/Src/RhoA/ROCK/MKK3/6/p38 pathway is fully activated downstream of α3β1 in EC, endothelial ERK1/2 is progressively dephosphorylated despite time‐dependent MEK1/2 activation. We further demonstrate that ERK1/2 dephosphorylation results from a crosstalk between p38 and ERK1/2 pathways whereby p38‐dependent increase in the expression of dual specificity phosphatase MKP‐3 causes progressive ERK1/2 deactivation. The resulting outcome of this complex crosstalk between the two major endothelial MAP kinases is p38‐dependent activation of MT1‐MMP, increase in MMP‐9 expression, and progressive release of MMP‐2 from ECs. We conclude that upon interaction with ECs, metastatic cells highjack EC MMP system via α3β1‐induced crosstalk between major MAPK pathways to facilitate tumor cell extravasation.Grant Funding Source: Supported by the VA BLR&D Service Award 1I01BX000609 and NIH Grant R01CA160461 to VV Glinsky
High glucose stress induces mesenchymal stromal cell senescence through up‐regulating autophagy (LB842)
Authors: Chang Tzu‐Ching , Hsu Min‐Fen , Wu Kenneth K. 
Published: June 2021

Diabetic hyperglycemia was reported to induce impairment of human bone marrow (BM) hematopoietic function and cause niche dysfunction. As multi‐potent mesenchymal stromal cells (MSC) are a key constituent of BM hematopoietic niche, we determined the influence of high glucose (HG) on cultured MSCs. MSCs were cultured in medium containing 5.5 mM glucose (low glucose, LG) or 25 mM glucose (HG) for up to 28 days and cell proliferation and morphology were analyzed. MSCs cultured under HG exhibited reduced cumulative population doubling and BrdU incorporation and increased p16 and p21 in a time‐dependent manner. Senescence‐associated β‐galactosidase (SA‐β‐Gal) positive cells and interleukin‐6 (IL‐6) production were increased. Cells underwent morphological changes consistent with senescence. Since cellular senescence is controlled by autophagy, we determined whether HG suppresses autophagy thereby inducing premature senescence. Interestingly, MSC cultured in HG medium exhibited enhancement of beclin‐1 expression and LC3‐II generation when compared to MSC cultured in LG. HG increased beclin‐1 (Atg 6) as well as Atg 5, Atg 7, and Atg 12 mRNA when compared to LG. HG also significantly increased autophagosome formation. To elucidate the relationship between autophagy and senescence, we treated MSCs with 3‐methyladenine (3‐MA), an inhibitor of autophagy and analyzed changes in senescence. 3‐MA prevented growth arrest and p21 elevation and abrogated SA‐β‐Gal and IL‐6 expressions. These results indicate that HG induces senescence through activation of autophagy. It was reported that HG induces cell injury via generation of reactive oxygen species (ROS). We evaluated the effect of N‐acetylcysteine (NAC), an antioxidant, on MSC autophagy and senescence. NAC reduced Atg expressions, LC3‐II generation and autophagosome formation accompanied by preventing senescence. DPI, an inhibitor of NADPH oxidase, exerted a comparable effect as NAC.These findings indicate that HG induces premature senescence by ROS‐mediated autophagy upregulation. MSC senescence contributes to niche dysfunction and bone marrow inflammation.
Virtual Humans and Avatars ‐ Healthcare Personal Agents in the Palm of Your Hand (LB855)
Authors: Federico Niksha , Federico Tessya , Federico Francesca , Sternik Gabriel , Esmaeli‐Azad Babak 
Published: June 2021

The advent of wireless sensors and miniaturized wearables have made the possibility of home diagnostics prior to a doctor visit and connected healthcare a reality. Engagement of everyday people for self diagnosis, however, remains a continued challenge as in most cases the much needed user friendly and engaging user interface for wireless devices are lacking.Recent advancements in the fields of physiology, psychology, artificial intelligence, speech recognition, natural language learning, computer graphics, gamification and cinematography have made the development of virtual human/ avatar personal health agents that are engaging, wireless smart phone and tablet native, and personalized possible.We here describe development of a personalized virtual human/ avatar health agent for incentivized, gamified, home self testing of 21 indications using FDA approved miniaturized wearable medical diagnostic devices. The avatars are designed to have multiple characters capable of understanding 15 different languages. Data obtained from patient interactions and device testing is stored in the cloud for analysis. Aesthetics of patient engagement and gamification will be presented. Development of disease specific scripts to determine different stages of a particular disorder including live physician telemedicine capabilities and connected healthcare will be described. Our findings are used as a solution for the development of team SMART McCoy's solution for the Qualcomm Medical Tricorder X Prize competition. In full development this system will assist in lessening the unnecessary primary care doctor visits (i.e., more than $10 billion annual cost in US alone) and provides the mean for making connected healtcare and precision medicine a reality.
Metabolics of PH – an update
Authors: Hughes Allyson L. , Hemnes Anna R. 
Published: June 2021

Purpose of review
While there has been a longstanding interest in metabolic disease in pulmonary hypertension, publications in the last several years have translated basic science findings to human disease and even led to recently published studies of metabolic therapy in pulmonary arterial hypertension that are discussed here.

Recent findings
Progress has been made in four key areas including mechanisms of insulin resistance in pulmonary arterial hypertension, the role of obesity in pulmonary vascular disease, novel clinical trials targeting metabolism in pulmonary hypertension, and the role of metabolism in chronic thromboembolic pulmonary hypertension.

Summary
: Insulin resistance in pulmonary arterial hypertension is primarily in the lipid axis. There are systemic manifestations of insulin resistance including right ventricular lipotoxicity. Obesity is associated with elevation of right ventricular systolic pressure even in a healthy population and therapies in pulmonary arterial hypertension that target metabolism hold promise for improving exercise, right ventricular function, and visceral adiposity. Finally, there are emerging data that chronic thromboembolic pulmonary hypertension is similarly characterized by metabolic alterations, though the specific metabolites may be different from pulmonary arterial hypertension.
Urine Test Predicts Kidney Injury and Death in COVID-19
Authors: Xu Katherine , Shang Ning , Levitman Abraham , Corker Alexa , Kudose Satoru , Yaeh Andrew , Neupane Uddhav , Stevens Jacob , Mohan Sumit , Sampogna Rosemary , D’Agati Vivette , Kiryluk Krzysztof , Barasch Jonathan 
Published: June 2021

AbstractBackgroundKidney injury is common in COVID-19 infection, but serum creatinine (SCr) is not a sensitive or specific marker of kidney injury. We hypothesized that molecular markers of tubular injury could diagnose COVID-19 associated kidney damage and predict its clinical course.MethodsThis is a prospective cohort study of 444 consecutive COVID-19 patients (43.9% females, 20.5% African American, 54.1% Latinx) in Columbia University’s Emergency Department at the peak of the New York pandemic (March-April 2020). Urine and blood were collected simultaneously at admission (median time of day 0, IQR 0-2 days) and within 1 day of a positive SARS-CoV-2 test in 70% of patients. Biomarker assays were blinded to clinical data.ResultsUrinary NGAL (uNGAL) was strongly associated with AKI diagnosis (267±301 vs. 96±139 ng/mL, P=1.6×10−10). uNGAL >150ng/mL had 80% specificity and 75% sensitivity to diagnose AKIN stage 2 or higher. uNGAL quantitatively predicted the duration of AKI and outcomes, including death, dialysis, shock, and longer hospital stay. The risk of death increased 73% per standard deviation of uNGAL [OR (95%CI): 1.73 (1.29-2.33), P=2.8×10−4] and was independent of baseline SCr, co-morbidities, and proteinuria [adjusted OR (95%CI): 1.51 (1.10-2.11), P=1.2×10−2]. Proteinuria and uKIM-1 also indicated tubular injury but were not diagnostic of AKI. Typically, distal nephron segments transcribe NGAL, but in COVID-19 biopsies with widespread acute tubular injury (ATI), NGAL expression overlapped KIM-1 in proximal tubules.ConclusionuNGAL predicted diagnosis, duration, and severity of AKI and ATI, as well as hospital stay, dialysis, shock, and death in patients with acute COVID-19.
SARS-CoV-2 shedding, infectivity and evolution in an immunocompromised adult patient
Authors: Mendes-Correa Maria Cassia , Ghilardi Fábio , Salomão Matias Chiarastelli , Villas-Boas Lucy S. , Vicente de Paula Anderson , Tozetto-Mendoza Tania Regina , Freire Wilton , Sales Flavia C. , Romano Camila M. , Claro Ingra M. , de Souza Leandro Menezes , Ramos Jessica F. , Paiao Heuder Gustavo Oliveira , Szor Roberta Shcoinik , Kallas Esper Georges , Sabino Ester C. , Witkin Steven S. , Faria Nuno R. 
Published: June 2021

AbstractThis report describes a persistent SARS-CoV-2 infection of at least 218 days in a male, in his 40’s who had undergone a prior autologous hematopoietic stem cell transplant due to a diffuse large B-cell lymphoma. He did not manifest a humoral immune response to the virus. Whole-genome sequencing and viral cultures confirmed a continual infection with a replication-positive virus that had undergone genetic variation for at least 196 days following symptom onset.
Natural language inference for clinical registry curation
Authors: Percha Bethany , Pisapati Kereeti , Gao Cynthia , Schmidt Hank 
Published: June 2021

AbstractClinical registries - structured databases of demographic, diagnosis, and treatment information for patients with specific diseases or phenotypes - play vital roles in high-quality retrospective studies, operational planning, and assessment of patient eligibility for research, including clinical trials. However, registries are extremely time and resource intensive to curate. Natural language processing (NLP) can help, but standard NLP methods require specially annotated training sets or the construction of separate models for each of dozens or hundreds of different registry fields, rendering them insufficient for registry curation at scale. Natural language inference (NLI), a specific branch of NLP focused on logical relationships between statements, presents a possible solution, but NLI methods are largely unexplored in the clinical domain outside the realm of conference shared tasks and computer science benchmarks. Here we convert registry curation into an NLI problem, applying five state-of-the-art, pretrained, deep learning based NLI models to clinical, laboratory, and pathology notes to infer information about 43 different breast oncology registry fields. We evaluate the models’ inferences against a manually curated, 7439 patient breast oncology research database. The NLI models show considerable variation in performance, both within and across registry fields. One model, ALBERT, outperforms the others (BART, RoBERTa, XLNet, and ELECTRA) on 22 out of 43 fields. A detailed error analysis reveals that incorrect inferences primarily arise through models’ misinterpretations of temporality--they interpret historical findings as current and vice versa--as well as confusion based on subtle terminology and abbreviation variants common in clinical notes. However, modern NLI methods show promise for increasing the efficiency of registry curation, even when used “out of the box” with no additional training.
A pilot feasibility study on SARS-CoV-2 detection method based on nasopharyngeal lavage fluid
Authors: Frezza Daniele , Fabbris Cristoforo , Franz Leonardo , Vian Elisa , Rigoli Roberto , De Siati Rosalba , Emanuelli Enzo , Luigi Bertinato , Boscolo-Rizzo Paolo , Spinato Giacomo 
Published: June 2021

AbstractObjectiveNose and nasopharyngeal swab is the preferred and worldwide accepted method to detect the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) within the nose and nasopharynx. This method may be linked with possible difficulties, such as patient’s discomfort or complications. This paper shows a pilot study of SARS-CoV-2 detection with nasal and nasopharyngeal lavage fluids.MethodsNasal lavage fluid was collected from patients who were submitted to SARS-CoV-2 screening test, due to a preceding positive rapid antigen test. A control group was enrolled among healthcare professionals whose nasopharyngeal swab tested negative. Nasal lavages were performed using isotonic saline solution injected through a nasal fossa. Both lavage fluid and traditional nasopharyngeal swab were analyzed by real-time PCR and antigenic test.ResultsA total of 49 positive subjects were enrolled in the study. Results of the analysis on lavages and nasopharyngeal swabs were concordant for 48 cases, regardless of the antigenic and molecular test performed. RT-PCR resulted weakly positive at swab in one case and negative at lavage fluid. Among the control group (44 subjects) nasopharyngeal swab and lavage fluid analyses returned a negative result. Sensitivity of the molecular test based on nasal lavage fluid, compared to traditional nasal swab, was 97.7%, specificity was 100%, and accuracy was 98.9%, with high agreement (Cohen’s k, 0.978).ConclusionNasal and nasopharyngeal lavages resulted to be highly reliable and well tolerated. A larger series is needed in order to confirm these results. This approach may potentially represent a valid alternative to the traditional swab method in selected cases.
A palmitoylation code controls PI4KIIIα complex formation and PI(4,5)P2 homeostasis at the plasma membrane
Authors: Batrouni Alex G. , Bag Nirmalya , Phan Henry , Baird Barbara A. , Baskin Jeremy M. 
Published: June 2021

ABSTRACTPI4KIIIα is the major enzyme responsible for generating the phosphoinositide PI(4)P at the plasma membrane. This lipid kinase forms two multicomponent complexes, both including a palmitoylated anchor, EFR3. Whereas both PI4KIIIα complexes support production of PI(4)P, the distinct functions of each complex and mechanisms underlying the interplay between them remain unknown. Here, we present roles for differential palmitoylation patterns within a tri-Cys motif in EFR3B (Cys5/Cys7/Cys8) in controlling the distribution of PI4KIIIα between these two complexes at the plasma membrane and corresponding functions in phosphoinositide homeostasis. Spacing of palmitoyl groups within three doubly palmitoylated EFR3B “lipoforms” affects both its interactions with TMEM150A, a transmembrane protein governing formation of a PI4KIIIα complex functioning in rapid PI(4,5)P2 resynthesis following PLC signaling, and its partitioning within liquid-ordered and -disordered regions of the plasma membrane. This work identifies a palmitoylation code in controlling protein–protein and protein–lipid interactions affecting a plasma membrane-resident lipid biosynthetic pathway.SUMMARY STATEMENTDifferent palmitoylation patterns on a lipid kinase adaptor protein control partitioning of the kinase between two spatiotemporally and functionally distinct complexes within the plasma membrane.
Expanding Clinical Phage Microbiology: Simulating Phage Inhalation for Respiratory Tract Infections
Authors: Porat Shira Ben , Gelman Daniel , Yerushalmy Ortal , Alkalay-Oren Sivan , Coppenhagen-Glazer Shunit , Cohen-Cymberknoh Malena , Kerem Eitan , Amirav Israel , Nir-Paz Ran , Hazan Ronen 
Published: June 2021

AbstractPhage therapy is a promising antibacterial strategy for resistant respiratory tract infections. Phage inhalation may serve this goal; however, it requires a careful assessment of their delivery by this approach. Here we present anin-vitromodel to evaluate phage inhalation.Eight phages, most of which target CF-common pathogens, were aerosolized and administered to a real-scale CT□derived 3D airways model with a breathing simulator. Viable phage loads reaching the output of the nebulizer and the tracheal level of the model were determined and compared to the loaded amount.Phage inhalation resulted in a diverse range of titer reduction, primarily associated with the nebulization process. No correlation was found between phage delivery to the phage physical or genomic dimensions. These findings highlight the need for tailored simulations of phage delivery, ideally by a patient-specific model in addition to proper phage matching, to increase the potential of phage therapy success.Take-Home MessagePhage therapy can be used against infectious diseases if personally tailored. Using a 3D airways model, we show that phage delivery by inhalation to the respiratory tract is unpredictable and also requires a precise evaluation.
OsciDrop: A Versatile On-demand Droplet Generator
Authors: Ye Shun , Zheng Xu , Li Caiming , Huang Weihang , Tao Yi , Yu Yanghuan , Yang Limin , Lan Ying , Ma Liang , Bian Shengtai , Du Wenbin 
Published: June 2021

ABSTRACTDroplet microfluidics is a powerful tool in many biological and clinical applications. Microfluidic chips, such as flow-focusing droplet generators, have been extensively used to high-throughput encapsulate reactions with single-cell and single-molecular resolutions. However, microfabrication is expensive and precision-demanding, preventing it from widespread use in biomedical laboratories and clinical facilities. Herein, we present a versatile chip-free droplet generator, OsciDrop, for generating size-tunable droplets on demand, with high uniformity. OsciDrop segments the fluid flowing out of the orifice of a micropipette tip into droplets by oscillating the tip under the surface of a continuous oil phase. We investigated the factors influencing droplet generation by examining several control parameters. Results show that flow rate, oscillating amplitude, and frequency are key parameters to generate monodisperse droplets on demand. And OsciDrop is able to generate droplets in a flexible and repeatable manner. Importantly, using an optimal asymmetrical oscillation waveform, OsciDrop can controllably generate monodisperse droplets spanning a wide volume range (200 pL - 2 μL). To demonstrate the ability of OsciDrop for chip-free droplet assays, a digital loop-mediated isothermal amplification (dLAMP) was performed to absolutely quantify African swine fever virus (ASFV). The OsciDrop method opens up a feasible and versatile avenue to perform droplet-based assays, exhibiting full accessibility for chip-free droplet microfluidics.