bluuebunny/crossref_metadata_2025_split huggingface dataset:
Deletion of Kir5.1 Abolished the Inhibitory Effect of High Sodium (HS) Intake on the Basolateral K Channels in the DCT and Thiazide‐Sensitive Na‐Cl Cotransport (NCC)
Authors: Lin Daohong , Wu Peng , Zhang Dandan , Gao Zhong-Xiuzi , Duan Xinpeng , Xiao Yu 
Published: April 2020

BackgroundWe have previously demonstrated that dietary sodium intake regulated the basolateral Kir4.1/Kir5.1 channel activity in the distal convoluted tubule (DCT) such that low sodium (LS) intake stimulated whereas HS intake inhibited the Kir4.1/Kir5.1 in the DCT. The HS‐induced inhibition of basolateral Kir4.1/Kir5.1 in the DCT was essential for the HS‐induced inhibition of NCC because HS failed to inhibit NCC in the kidney‐specific Kir4.1 knockout mice. While Kir4.1 is responsible for permeating the basolateral K conductance, Kir5.1 is an important regulatory subunit of the basolateral K channel of the DCT. The aim of the present study is to explore the role of Kir5.1 in mediating the effect of HS on the basolateral K conductance of the DCT and NCC expression/activity.MethodsWe have used the whole‐cell recording technique to measure the basolateral K (Kir4.1) conductance and DCT membrane potential, employed immunoblotting to examine the expression of phosphor‐NCC (pNCC) and total NCC (tNCC), and conducted renal clearance method to measure thiazide‐sensitive renal sodium excretion in Kcnj16+/+ (WT) mice and Kcnj16−/− mice (Kir5.1 KO).ResultsWe confirmed the previous finding that HS decreased the basolateral K conductance of the DCT, depolarized the DCT membrane, and inhibited the expression of pNCC and tNCC in WT mice. In contrast, LS intake increased the basolateral K conductance, hyperpolarized DCT membrane, and augmented expression of pNCC and tNCC in WT mice. The deletion of Kir5.1 not only increased the basolateral K (Kir4.1) conductance in the DCT and also abolished the inhibitory effect of HS on the basolateral Kir4.1 activity. Also, LS intake did not further stimulate the basolateral Kir4.1 of the DCT. The expression of pNCC and tNCC was upregulated in Kir5.1 KO mice in comparison to WT. Furthermore, neither HS nor LS had an effect on NCC expression in Kir5.1‐deficient mice, suggesting the role of Kir5.1 in mediating the inhibitory effect of HS on NCC expression. Renal clearance experiments also showed that the deletion of Kir5.1 not only augmented the thiazide‐induced natriuresis but also abolished the inhibitory effect of HS on the thiazide‐induced natriuresis in Kir5.1 KO mice.ConclusionKir5.1 plays an important role in mediating the inhibitory effect of HS intake on the basolateral K channel activity in the DCT and NCC activity/expression.Support or Funding InformationNIH‐DK115366
The Oligomerization Status of IRE1α Kinase/RNase Determines Its Signaling Outputs Under Endoplasmic Reticulum Stress
Authors: Oakes Scott A. 
Published: April 2020

Accumulation of unfolded proteins in the endoplasmic reticulum (ER) triggers signaling pathways called the unfolded protein response (UPR). Under remediable levels of ER stress, the adaptive UPR (A‐UPR) activates transcriptional and translational changes that restore homeostasis. However, under irremediably high ER stress, these adaptive measures fail and the signaling pathways instead trigger programmed cell death—referred to as the terminal UPR (T‐UPR). We discovered that IRE1α—an ER transmembrane bifunctional kinase/endoribonuclease (RNase)—is a critical life‐death switch that employs kinase auto‐phosphorylation levels as a mechanism to control its filamentous status and toggle between an A‐UPR or a T‐UPR depending on the level of upstream ER stress. Remediable ER stress causes low‐level kinase auto‐phosphorylation and dimerization that restricts IRE1α’s RNase activity to excising an intron in XBP1 mRNA, which upon re‐ligation and translation of spliced XBP1 produces the homeostatic transcription factor XBP1s (s=spliced) that then upregulates genes encoding ER protein‐folding and quality control components. However, sustained ER stress induces high‐level kinase autophosphorylation to promote higher‐order IRE1α oligomerization; under these conditions, IRE1α RNase hyperactivation leads to massive degradation of ER‐localized mRNA and T‐UPR events including: (1) loss of differentiated cell identity, (2) local sterile inflammation, and (3) programmed cell death through pyroptosis and apoptosis. We have recent insights into how the oligomerization status and signaling outputs of IRE1α contribute to cancer.Support or Funding InformationThe work was supported by grants: American Cancer Society Research Scholar Award; Harrington Discovery Institute Scholar‐Innovator Award; UCSF and Onyx Oncology Innovation Alliance; American Association of Cancer Research; R01CA219815; R01EY027810.
Pembrolizumab treatment increases chemotaxis of cytotoxic T cells in head and neck cancer patients within the adenosine‐rich tumor microenvironment
Authors: Newton Hannah S. , Chimote Ameet A. , Gawali Vaibhavkumar S. , Palackdharry Sarah , Hildeman David , Janssen Edith , Wise-Draper Trisha , Conforti Laura 
Published: April 2020

Head and neck cancer (HNC) is the sixth most common cancer worldwide with a five‐year survival rate of 50%. Immunotherapy has been emerging as a promising treatment modality as conventional treatment strategies (surgery, chemotherapy and radiation) have limited efficacy and cause debilitating co‐morbidities. Immunotherapy harnesses the patient’s own immune system to fight disease. Targeting the immune system is crucial because immune cell (specifically cytotoxic T cell) infiltration into the tumor microenvironment (TME) and subsequent cytotoxicity is critical for tumor elimination. Pembrolizumab, an anti‐PD‐1 (programmed cell death‐1 receptor) antibody, is an immunotherapy currently approved for metastatic or recurrent unresectable HNC and is in clinical trials for curative intent. While there have been promising long‐lasting clinical responses to Pembrolizumab, many patients still do not respond. Thus, it is critical to understand the mechanism of action of Pembrolizumab in order to reduce treatment resistance. Moreover, it has been shown that Pembrolizumab increases cytotoxic T cell density in the TME; nevertheless, the underlying mechanism is not fully understood. However, it is known that there is an increased accumulation of adenosine (Ado), an anti‐inflammatory purine nucleoside, in the TME of HNC patients, which inhibits T cell chemotaxis by blocking the function of the potassium ion channel, KCa3.1. Therefore, to elucidate possible effects of Pembrolizumab on T cell chemotaxis, we studied KCa3.1 activity and subsequent chemotaxis in CD8+ peripheral blood T cells (PBTs) of naïve (never treated) HNC patients before/after Pembrolizumab treatment (n=25) via electrophysiological and 3D chemotaxis experiments. Furthermore, Pembrolizumab treated patients were categorized as responders (R) or non‐responders (NR) based on treatment response (i.e., reduction in tumor viability with a cutoff of ≥20% for R). We observed that Pembrolizumab increased the KCa3.1 activity in post‐treatment patient PBTs by 29.8% (p=0.026). Because we observed an increase in KCa3.1 activity in treated patients, we next assessed whether there was an effect on the chemotaxis of PBTs in presence of Ado. While Ado inhibited the chemotactic ability in treatment naïve PBTs by 53.9% (p<0.001, n=22), we determined that post‐treatment R PBTs were not inhibited by Ado (p = 0.142, n=11) whereas post‐treatment NR PBTs were inhibited by Ado (p = 0.005, n=10). These data support a role of ion channels and chemotaxis in patient response to Pembrolizumab and could lead to better understanding of the effect of Pembrolizumab in immune cell infiltration into the tumor and, ultimately, its mechanism of action/resistance.Support or Funding InformationDoD CA160714; T32CA117846; 2R01 CA95286
Pharmacologic Effects of a Synthetic Macamide Analogue (ZYPC) on A549 and HeLa Cells
Authors: Lam Melanie , Rondón-Ortiz Alejandro Nico , Martínez-Málaga Jimena Alexandra , Gonzales-Urday Ana Lucia , Pino-Figueroa Alejandro 
Published: April 2020

Lepidium meyenii L., known as the Maca plant, is acclaimed by the scientific community for its various health benefits in traditional medicine, including prevention of gastritis, depression, and cancer, as well as increasing fertility and improving sexual performance. Macamides (N‐benzylamides) are secondary metabolites derived from Maca that have demonstrated neuroprotective, anti‐oxidative, anti‐inflammatory, endocannabinoid system modulatory, and mild anti‐diabetic properties. The present study investigates the pharmacologic effect of N‐(8Z‐heptadecen‐1‐yl)‐O‐(3‐pyridylmethyl) carbamate (ZYPC), a synthetic macamide analog, on A549 non‐small cell lung cancer and HeLa cervical cancer cells. Both cell lines were treated with different concentrations of ZYPC (1–100 μM). After 24 h, cell viability was measured using a 3‐(4, 5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTS) colorimetric assay. ZYPC was found to significantly reduce cell viability in a dose‐dependent manner. The half maximal inhibitory concentration was calculated, using dose‐response non‐linear regression, as 38.94 μM for A549 cells and 38.35 μM for HeLa cells. Subsequently, a caspase 3 colorimetric assay was executed to study ZYPC‐induced reduction of cell viability through activation of apoptosis. HeLa cells were treated with 0, 30, 50, and 100 μM ZYPC and cell lysates were collected after 5 h incubation. A reagent containing DEVD‐pNA, a substrate for caspase 3, was added to the lysates, and incubated for 2 h at 37°C. Unexpectedly, ZYPC was found to have no effect on caspase 3 activity. Then, the involvement of a pro‐survival mechanism was assessed through AKT and phospho‐AKT (p‐AKT) chemiluminescent western blotting. Results obtained from treatment with 0, 10, 30, and 50 μM ZYPC for 24 h were inconclusive for total AKT and p‐AKT levels in both cell lines. Preliminary western blots of poly (ADP‐ribose) polymerase (PARP), a DNA repair enzyme, and its inactive cleaved form indicated constant expression levels of both proteins in A549 cells. Interestingly, there was an apparent increase in cleaved PARP in HeLa cells, which may suggest late‐stage apoptosis. However, further studies are needed to better elucidate the mechanism of anti‐cancer activity exhibited by ZYPC.Support or Funding InformationMCPHS University Summer Undergraduate Research Fellowship Program
Balancing Parts: Integrating Uncertainty/Ambiguity into Anatomy Education
Authors: Lazarus Michelle D. , Rees Charlotte , Stephens Georgina 
Published: April 2020

Tolerance of Uncertainty/Ambiguity (ToU/A) is a construct describing individuals’ responses to ambiguous stimuli. ToA is recognised as impacting healthcare outcomes, however, little is known about whether education may impact students’ ToA, and if so, how we can develop pedagogy to improve students’ ToU/A. Framework analysis of longitudinal qualitative data, collected via online discussions and semi‐structured interviews from 207 undergraduate medical students across three semesters of anatomy, identified anatomy pedagogy which either fostered or hindered students’ ToU/A. Anatomy curricula which integrates opportunities for students to distribute responsibility for knowledge (i.e. working in teams) and/or provide anonymity for those conveying knowledge (i.e. online anonymous discussion forums) appear to foster students’ ToU/A, while didactic educator approaches (suggesting that knowledge is finite and knowable) appear to hinder students’ ToU/A. Thematic analysis across time‐points suggest that active learning approaches can be effective stimuli for students’ ToU/A development. Active learning approaches within our study balanced communication of anatomy discipline knowledge with clinical applications, so that the context of knowledge became the stimulus of uncertainty. Exemplar anatomy curricula fostering ToU/A include “choose your own adventure” chest pain team‐based learning, where students elect which patient information they receive and in which order, engaging anatomical knowledge through the “adventure”. Another approach is using short clinical case scenarios wherein more than one underlying anatomical deficit may explain symptoms described in the scenario. Despite evidence that students develop ToA/U in their cognition and behaviour (e.g. through acknowledgement of the clinical importance of ToU/ToA, and perseverance when faced with ambiguous/unknown knowledge), student talk in this study suggests that emotionally, students remain negative about ToU/A (e.g. feeling overwhelmed, fearing errors). Together our data and examples illustrate that foundational science knowledge education can be balanced with effective student development of ToU/A, but student feedback may remain negative; an important consideration in evaluating effective curricula. Future work will focus on whether students’ negative emotions change with time and clinical exposure.Support or Funding InformationResearch supported by Monash Education Academy, Faculty of Medicine, Nursing and Health Sciences (Monash University) and a Medical Education Travel Fellowship
Whitening Effect of Hydrothermal Extracts of Phragmites Communis and its Active Compound, p‐coumaric acid
Authors: So Gyeongseop , Ha Chang Woo , Jang Sohee , Kim Sung Hyeok , Kim Youn Kyu , Lim Hyosun , Sohn Eun-Hwa 
Published: April 2020

Reed (Phragmites communis) is a perennial grass forming extensive colonies inwetland. The study aimed to compare the antioxidant and whitening effects of roots of P. communis extracts (RPC) by three different extraction processes, using 100% EtOH, 70% EtOH and hydrothermal fluid methods. The amounts of total phenolic compound(TPC), total flavonoid contents(TFC), and p‐coumaric acid (pCA), a major active constituent, were evaluated. Melanogenesis‐related factors such as tyrosinase, L‐DOPA oxidase, and DPPH radical scavenging activities by all three RPC were also measured. Among them, hydrothermal method of RPC(h‐RPC) showed the best effect in the contents of TPC and TFC, DPPH scavenging activity, tyrosinase‐ and L‐DOPA oxidase inhibitory activities. Highest pCA content was also detected from h‐RPC (3.05 ppm). Finally, we confirmed that the color of the cell pellet and the amount of melanin synthesis were significantly reduced in the presence of h‐RPC and pCA in α‐melanin cell stimulating hormoneinduced B16F10 cells. The anti‐melanogenic potential of h‐RPC and pCA will provide an insight into developing a new skin whitening product.Support or Funding InformationThis research was a part of the project entitled "Development of multi‐functional cosmetic materials derived from halophyte", funded by the Ministry of Oceans and Fisheries, Korea.
Comparing in vitro and in vivo Bicistronic Design Protein Production
Authors: Gomez Paul , McKlin Sam , Chen Queena , Mazariegos Christopher , Zhou John , Seligmann Haley , Bashaw Benjamin , Peterson Andy , Shildknecht Kaitlyn , Tauchen Ali , Tittor Raechel , Poet Jeffrey L. , Eckdahl Todd T. , Heyer Laurie J. , Campbell A. Malcolm 
Published: April 2020

Synthetic biology takes an engineering approach to biological systems for the construction of parts, devices and systems that address challenges associated with medicine, biofuels, the environment, and more. Although the production of natural and engineered proteins is fundamental to this effort, a persistent challenge to protein production has been unreliable translation directed by a given ribosomal binding site (RBS) depending on which gene of interest is downstream of it. Based on many observations, investigators have hypothesized that the 5’ UTR containing an RBS and early codons can bind to downstream coding mRNA to form base paired secondary structures that decrease the efficiency of translation initiation. In 2013, Drew Endy and colleagues developed bicistronic designs (BCDs) for predictable and reliable in vivo translation. BCDs contain two RBSs and two protein‐encoding cistrons. The first RBS leads to the production of a 16 amino acid leader polypeptide with no functionality, whereas the second RBS directs the production of a protein encoded by a gene of interest. A key feature of BCDs is that the stop codon for the first cistron (TAA) shares its terminal adenine with the first base of the start codon (ATG) for the second cistron. In other words, TAATG is both a stop codon in one reading frame of cistron 1 and a start codon in a shifted reading frame for cistron 2. BCDs were found to be better at producing predictable protein levels in vivo than simple RBSs, thereby improving the ability of BCDs to function reliably and predictably as classic synthetic biology parts in bacterial cells. We wanted to learn if BCDs worked equally well for in vitro protein production during cell‐free protein synthesis (CFPS). CFPS employs transcriptional and translational machinery extracted from cells producing the viral T7 RNA polymerase. We built GFP expression cassettes using 19 BCDs and tested them with CFPS. We compared the rank order of the 19 BCDs in terms of in vitro CFPS GFP production with the in vivo results published by the Endy group. We calculated a rank correlation of 0.88 between the in vitro and in vivo data, showing that BCD control of protein production is comparable in both contexts. The significance of our work is that synthetic biologists using CFPS are well advised to employ BCDs so they can produce predictable amounts of protein for widespread applications in vitro.Support or Funding InformationNSF RUI MCB‐1613281 to Missouri Western State University; NSF RUI MCB‐1613203 to Davidson College
Fundamentals of Teaching in the Basic Sciences
Authors: Goodman Barbara E. 
Published: April 2020

Graduate students in the School of Medicine have had an elective course on how to teach offered either annually or every other year since 2007. The initial course design was based on suggestions from other educator colleagues (especially Dr. Dee Silverthorn). The course is offered in 90 minute to 2 hour time blocks once a week throughout a 15‐week semester. The course is discussion‐based with online reading material provided through the course management system and various assignments during the course (mostly involving having each student design aspects of a hypothetical course that they may teach in their future careers). Topics for the discussions include: the three‐legged stool, importance of learning objectives, planning a course, designing a course, learning preferences, teaching styles, visual aids and active learning, assessing student learning, exams and grading, evaluating a course, teaching portfolios and other resources. The professors provide feedback for specific assignments during the course which include: a minute paper on a course that the student liked and why, how to choose a textbook, writing learning objectives, writing a syllabus, writing a report on a class observation, writing a short review paper on an evidence‐based teaching pedagogy, writing 5 active learning ideas for their course, a short videotaped teaching session (as a preliminary to the final microteach session), finding 3 different online resources for their course, and a peer and self‐evaluated 10‐minute videotaped microteach session that acts as the final examination for the course. The short review paper was added recently when this course became a required course for the non‐thesis option master’s program in basic biomedical sciences so that the students would have experience with scientific writing. The culminating videotaped microteach session was felt to be so valuable for the students that a short videotaped session was added earlier so that students could make improvements after their initial attempt. An interesting recent complication in the course is that although the course was initially designed for preparing future faculty, now the majority of the students in the course are pre‐professional students hoping for acceptance into health professional programs where classroom teaching may not be a high priority. Thus, the emphases of the discussions and assignments have changed to address relevance to these students. However, student evaluations following the two classes with predominantly master’s students have still shown appreciation of the discussions and assignments for future health practitioners.Support or Funding InformationDr. Goodman was partially supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103443.
The relationship between Alzheimer's disease and vascular risk factors
Authors: Chen Xianghui , Zhang Weidong , Qiao Xiaohong , Li Youliang , Duan Hubin , Sun Pinglian 
Published: April 2020

ObjectiveTo study the relationship between Alzheimer's disease(AD) and vascular risk factors.Methods53 patients with AD were selected as the case group and 40 elderly patients with normal cognitive function as the control group. The distribution of hypertension, diabetes, abnormal lipid metabolism, cerebral infarction, carotid artery stenosis and plaque, carotid artery intimal thickening, intracranial blood flow velocity slowing, smoking and drinking history were analyzed. Logistic regression model was used to analyze the relationship between above factors and AD.ResultsHypertension, diabetes, blood lipid metabolism, cerebral infarction, neck vascular stenosis and plaque, carotid artery intimal thickening, slow cerebral artery blood flow between AD and the control group with significant difference (P < 0.05), while there was no significant differences in moking and drinking alcohol history between AD case group and the control group (P > 0.05).ConclusionAD is closely related to vascular risk factors. Hypertension, diabetes, abnormal lipid metabolism, cerebral infarction, carotid artery stenosis and plaque, carotid artery intimal thickening and intracranial blood flow velocity are independent risk factors for AD.Support or Funding InformationThis study was supported by Lvliang City Science and Technology Fundation Grant, Lvliang people‘s hospital Fundation Grant, This study was supported by the National Natural Science Foundation Youth Fund (30600637), China Postdoctral Science Foundation Grant (2019T120195), China Postdoctral Science Foundation Grant (2014M561207), the Shanxi Province Basic Research Program ( 2010021034‐4, 201601D011119), the Shanxi Scholarship Council of China (2011‐096 and Key Projects No.4), Department of Human Resources and Social Security of Shanxi province, The science and technology research projects of health department of Shanxi Province(2016D011119), The Teaching Reform Program of Shanxi Higher Education, The Education Reform Project of Postgraduate Students in Shanxi Province, The Education Reform Project of Shanxi Medical University.