bluuebunny/crossref_metadata_2025_split huggingface dataset:
Angiotensin II Represses Guanylyl Cyclase/Natriuretic Peptide Receptor‐A Gene Expression and Receptor Signaling and Function
Authors: Arise Kiran K. , Kumar Prerna , Samivel Ramachandran , Zhao Hanqing , Lindsey Sarah , Pandey Kailash N. 
Published: April 2020

Atrial and brain natriuretic peptides (ANP and BNP) bind to guanylyl‐cyclase/natriuretic peptide receptor‐A (GC‐A/NPRA) and regulate sodium excretion, water balance, vasorelaxation, and steroidogenesis; all directed to combat hypertension. ANP acting through NPRA lowers blood pressure and blood volume and protects the heart and vascular disorders and end‐organ damage. The two vasoactive hormones, angiotensin II (ANG II; vasoconstrictive) and atrial natriuretic peptide (ANP; vasodilatory) antagonize the biological actions of each other. The objective of the present study was to examine whether ANG II plays critical roles in the transcriptional repression of Npr1 (encoding for NPRA) and receptor function. To delineate the mechanisms of ANG II‐mediated repression of Npr1 transcription and function, we performed the promoter deletional analyses, which showed that region −1182/−914 base pairs upstream from the transcription start site of the Npr1 promoter contains cis‐acting elements for ANG II‐regulated repression of Npr1. ANG II significantly decreased the levels of NPRA mRNA (60%), NPRA protein (52%), and intracellular accumulation cGMP (65%) in cultured mesangial cells and attenuated ANP‐mediated relaxation of aortic rings ex vivo. The results showed that transcription factors, CREB and HSF‐4a facilitated the ANG II‐mediated repressive effects on Npr1 transcription and receptor function. The current findings suggest that ANG II‐mediated repressive mechanisms of Npr1 transcription and receptor function may provide new molecular targets for the treatment and prevention of hypertension and renal and vascular disorders.Support or Funding InformationThis work was supported by NIH grant HL062147
ABO Blood Type and Von Willebrand Expression in Pulmonary Endothelial Cells
Authors: Post Ginell R. , Post Steven R. , Murray Glenn 
Published: April 2020

ABO blood group antigens are expressed on von Willebrand factor (VWF) and glycosylation patterns influence circulating VWF levels. Retrospective studies performed on patients with deep vein thromboses indicate that individuals with type O blood group have a decreased thrombotic risk secondary to lower von Willebrand factor (vWF) plasma levels compared to type A, B and AB (non‐O) individuals. We examined the effect of ABO blood type on tissue‐associated VWF protein and mRNA levels in formalin fixed paraffin‐embedded pulmonary tissue obtained at autopsy from decedents who died from pulmonary embolism with known ABO blood group phenotype (O, A, B or AB). Tissue microarrays were created and stained with antibodies to VWF and platelet/endothelial cell adhesion marker‐1 (PECAM‐1) as a marker of endothelial cells. A pixel count scoring algorithm was used to quantify VWF and PECAM‐1 staining intensity in pulmonary arterioles in digitized images. Compared with type O, non‐O individuals have a significantly higher amount of endothelial cell‐associated VWF protein expression. However, quantitate RT‐PCR showed that, when normalized to PECAM‐1, VWF transcripts were lower in type A individuals, compared to type O. In conclusion, VWF protein levels associated with pulmonary vascular endothelial cells are influenced by ABO antigenic determinants; however, protein levels are not regulated at the transcriptional level.Support or Funding InformationResearch was supported by funds provided by UAMS Department of Pathology.
Developmental loss of oligodendrocytes hinders adult CNS remyelination and increases astroglial and microglial activation
Authors: Pajoohesh-Ganji Ahdeah , Karl Molly , Garrison Eric , Clarkson-Paredes Cheryl , Miller Robert H. 
Published: April 2020

Multiple sclerosis, a neurodegenerative autoimmune disease characterized by loss of oligodendrocytes and myelin in the brain and spinal cord, results in localized functional deficits. Several risk factors have been associated with MS, however none can fully explain enhanced susceptibility to MS. Epidemiological data, based on geographical prevalence studies suggest susceptibility is established early in life and frequently long before disease diagnosis indicating developmental events influence adult disease progression. Here we test the hypothesis that loss of mature oligodendrocytes during postnatal development results in enhanced susceptibility to a second demyelinating insult. A transgenic mouse model was utilized to specifically induce apoptosis in a subset of mature oligodendrocytes (MBP‐iCP9) during the first 2 postnatal weeks followed by either a local LPC spinal cord injection or the induction of EAE in the adult. Immunostaining, immunoblotting, behavioral testing, and scanning electron microscopy were utilized to examine the differences between groups. We show that during development, oligodendrocyte apoptosis results in transient reductions in myelination and functional deficits that recover after 10–14 days. Compared to animals in which oligodendrocyte development was unperturbed, animals subjected to postnatal oligodendrocyte ablation showed delayed recovery from an LPC lesion. Unexpectedly, the induction and severity of EAE was not significantly altered in animals following oligodendrocyte ablation even though there was a substantial increase in spinal cord tissue damage and CNS inflammation. It is currently unclear why these changes are not reflected in enhanced functional deficits. These observations suggest that developmental loss of oligodendrocytes results in long lasting tissue changes that alter its capacity for repair in the adult.Support or Funding InformationNIH834857
Loss of AC10 impairs neutrophil recruitment into the lung following Pseudomonas aeruginosa infection
Authors: Sayner Sarah L. , Choi Chung-Sik 
Published: April 2020

Pseudomonas aeruginosa is a gram‐negative, opportunistic bacteria and a common cause of nosocomial pneumonia in critically ill patients that can progress to acute respiratory distress syndrome (ARDS). These patients have impaired gas exchange leading to hypoxia, altered blood pH and elevated bicarbonate. Bicarbonate, delivered into the cell via sodium bicarbonate cotransporters (NBCs), activates adenylyl cyclase 10 (AC10) to increase intracellular cAMP. Indeed, bicarbonate activation of AC10 is required for LPS‐induced lung endothelial permeability. Neutrophil recruitment into the airspaces plays a central role in the progression of ARDS. Recently, AC10 has been implicated in transendothelial emigration of neutrophils from the blood into the underlying tissue via CD99, a transmembrane protein critical for neutrophil recruitment. We sought to determine whether AC10 is necessary for neutrophil recruitment into the airspaces following P. aeruginosa intratracheal inoculation. Increasing doses of bacteria were introduced directly into the lung of wild type and AC10 and NBCn2 knockout mice. Bronchoalveolar lavage fluid (BALF) was recovered 24 hours later and examined for recruitment of polymorphonuclear cells (PMNs) into the airspaces. The number of PMNs recruited into the airspace following Pseudomonas inoculation increased with bacterial number and reached a maximum at a dose of 1E5 bacteria after which 100% of cells in the BALF were PMNs. Thus, doses upto 1E5 bacteria were used to examine the role of AC10 and NBCn2 in PMN recruitment into the airspace between wild type and knockout animals. Our data reveal a decreased number of PMNs in the BALF of both AC10 and NBCn2 knockout female mice; however, no difference in the number of PMNs recruited into the airspace was detected between male wild type and knockout animals. In addition, we confirmed the expression of CD99 in pulmonary microvascular endothelial cells and whole lung from both rat and mouse. Thus, CD99 is expressed in pulmonary endothelial cells and female mice utilize AC10 and NBCn2 to facilitate neutrophil recruitment into the airspace following a model of P. aeruginosa pneumonia.Support or Funding InformationNIH R01HL121513
Day‐to‐Day Reproducibility of the Near‐infrared Spectroscopy Venous Occlusion Technique in Young Healthy Adults
Authors: Dennis Joshua J. , Wiggins Chad C. , Smith Joshua R. , Isautier Jennifer M. J. , Johnson Bruce D. , Joyner Micheal J. , Cross Troy J. 
Published: April 2020

BackgroundPrevious studies have demonstrated that the near‐infrared spectroscopy venous occlusion technique (NIRS‐VOT) provides a valid assessment of regional forearm muscle blood flow; however, day‐to‐day reproducibility of hemodynamic variables derived from the NIRS‐VOT remains to be determined, particularly across the range of physiological blood flows observed during handgrip exercise.PurposeThe purpose of this study was to evaluate the day‐to‐day reproducibility of forearm hemodynamics assessed via the NIRS‐VOT.MethodsNear‐infrared spectroscopy (NIRS) data were obtained from the flexor digitorum superficialis muscle on the right arm of 14 young, healthy adults (age: 27 ± 6 yrs; BMI: 24.6 ± 2.8 kg·m−2). The NIRS‐VOT was performed via rapid inflation of a pneumatic cuff placed on the upper arm for approximately 10 s. The NIRS‐VOT was performed at rest, and immediately after a 15 s isometric handgrip contraction at 20%, 40% and 60% of the subject’s maximum voluntary contraction. Measurements were repeated 3–5 times. Participants returned to the laboratory for a second visit on a separate day, and repeated the same number of measurements. Forearm blood flow (FBF), and blood pressure were measured on a beat‐by‐beat basis, and forearm vascular conductance (FVC) was calculated. FBF and FVC were subsequently compared for day‐to‐day reproducibility.ResultsFBF and FVC significantly increased with the intensity of handgrip exercise (P< 0.05); however, these measures of forearm hemodynamics were not significantly different (P >0.05) between Day 1 and Day 2 at each exercise‐intensity (Table
).ConclusionThe findings of the present study indicate that the NIRS‐VOT provides a reliable assessment of regional forearm muscle hemodynamics across a physiological range of blood flows incurred by isometric handgrip exercise.Support or Funding InformationMJJ is funded by the NIH (R‐35‐HL139854) and the Mayo Foundation. CCW is supported by the NIH training grant (5T32DK007352‐39), JRS is supported by AHA (18POST3990251), and TJC is funded by AFAR Postdoctoral Transition Award (DIAMOND18161).

FBF and FVC on experimental days

Day 1

Day 2

P‐value

FBF, mL·min−1·100mL−1 (tissue)

Rest

2.0 ± 0.5

0.8 ± 0.2

0.001

20% MVC

14.4 ± 2.65

15.4 ± 2.4

0.679

40% MVC

26.0 ± 5.19

28.0 ± 4.3

0.606

60% MVC

39.4 ± 6.39

47.4 ± 7.7

0.858

FVC, mL·min−1·100 mmHg−1

Rest

170 ± 47.3

72 ± 16.4

0.001

20% MVC

1297 ± 227.8

1454 ± 209.6

0.380

40% MVC

2496 ± 492.8

2656 ± 434.0

0.912

60% MVC

3820 ± 659.1

4542 ± 744.7

0.535

MVC, kg

49.4 ± 3.0

47.9 ± 2.6

0.721

Abbreviations: FBF, forearm blood flow; FVC, forearm vascular conductance; MVC, maximal voluntary contraction. Values are presented as means ± SE. P values indicate comparison between days 1 and 2.
Tritiated JNJ‐63514230 is a novel GluN2B‐selective radioligand with no affinity to sigma receptors
Authors: Schoellerman Jeffrey , Lord Brian , Bhattacharya Anindya , Bonaventure Pascal , Letavic Michael , Lovenberg Tim , Balana Bartosz 
Published: April 2020

IntroductionCommonly used GluN2B radioligands exhibit limited selectivity towards GluN2B receptors as they interact with sigma and other off‐target receptors. Here we describe development and characterization of [3H] JNJ‐63514230, a novel radioligand selective for GluN2B‐containing NMDA receptors.MethodsThe selectivity JNJ 63514230 was determined using functional assays and CEREP panel. Saturation binding and competitive radioligand binding experiments were used to characterize [3H]‐JNJ 63514230 properties. We have also tested utility of [3H]‐JNJ 63514230 as a tracer for ex‐vivo autoradiography in rat brain.ResultsWe confirmed that widely used tracers [3H] Ifenprodil and [3H] Ro 25‐6981 have substantial affinity to sigma receptors as measured in radioligand competition experiments (CEREP). In contrast, new tracer [3H] JNJ‐63514230 exhibits no affinity to sigma 1 and sigma 2 receptors in CEREP assays. Saturation binding in rat brain membranes yielded a Kd value of 2.53 nM and Bmax of 2531 fmol/mg. Association constant Kon, dissociation constant Koff, and calculated equilibrium binding constant (Kon/Koff) were 0.0067±0.001 min−1 nM‐1, 0.021±0.0002 min−1, and 3.28±0.61 nM, respectively (average of 3 individual experiments each ± SD). Enhanced selectivity for GluN2B‐containing receptors translates to reduced non‐specific binding and better signal to noise ratio in both in vitro radioligand binding assays and ex vivo autoradiography.ConclusionsWe showed that new radioligand [3H] JNJ 63514230 is superior to previous previously described GluN2B tracers as it permits accurate measurement of affinity to GluN2B‐containing receptors (Ki) without using additional blockers to mask native sigma receptors.Support or Funding InformationAuthors are full time employees of Janssen Research & Development, LLC. The research was funded by the company.
SOARing Achievements: Do Medical Students Think Our New Teaching Strategy Achieved Its Goals?
Authors: Gritton Cory Buenting , Carmichael Heather , Kondo Kimi , Lee Lisa , Royer Danielle 
Published: April 2020

In 2018, the University of Colorado implemented Structured Obligatory Application & Review (SOAR) sessions designed around 11 specific goals associated with evidence‐based trends in medical education in order to improve the anatomy block. The sessions, spaced across the block, each included 4 rotations with small group work and hands‐on activities integrating clinical and anatomy content. Rotations included embryonic relationships with gross, anatomy in imaging & physical exam, surgical cases with cadaveric correlation, and team‐based learning of board‐style questions. The objective of this study was to evaluate whether SOAR successfully achieved its goals: peer‐to‐peer learning, self‐directed learning, identifying knowledge gaps, engaging review, long‐term retention, confidence in anatomy, connecting anatomy concepts, exam preparation, clinical relevance, and clinical application.In an IRB exempt study, 1st year medical students (n=182) were invited to complete a paper survey during the last SOAR in 2019. The survey included 11 Likert items pertaining to stated SOAR goals. Previously, 1st‐year students in the 2018 pilot SOAR (n=184) were invited to complete an online survey after block completion. The 2018 survey included the same 11 items, plus two free response questions that were qualitatively analyzed for themes.The 2019 survey response rate was 81% (n=149), much greater than 2018 online response rate (31%, n=59). Analysis of 2019 responses revealed student agreement greater than 50% for all 11 SOAR goals. Strong achievement of a goal was determined from upper quartile (75–100%) agreement ratings, which was seen for 4 goals: providing clinical relevance (91%), applying anatomical knowledge to clinical situations (89%), identifying knowledge gaps (82%), and connecting anatomical concepts (78%). Goals were considered moderately achieved with agreement ratings in the next quartile (50–74%), with the remaining 7 SOAR goals within this designation: an engaging way to review anatomy (70%), self‐directed learning (69%), peer‐to‐peer teaching (68%), preparation for written examinations (67%), long‐term retention (57%), improving confidence in anatomy (52%), and preparation for practical exams (51%). Cohort comparison revealed no significant difference in responses between the pilot and subsequent year of SOAR implementation for each of the goals (Mann Whitney U tests, p>0.05). Overall, SOAR goals were strongly to moderately achieved based on student perceptions, with benefits perceived strongest in providing clinical relevance, clinical application of anatomy, knowledge gap identification, and providing an engaging review. This suggests that developing targeted, interactive, small group activities such as those used in SOAR may be a valuable evidence‐based approach to better integrate clinical and anatomical content in the modern pre‐clinical medical curriculum.
Decreasing the survival ability of brain metastatic breast cancer cell by targeting glucose transporter 3
Authors: Kuo Min Hsun , Lee Hsueh-Te 
Published: April 2020

Breast cancer brain metastases (BBMs) are found in 15–20 % of breast cancer patients. And it results in a high mortality once the brain metastases are diagnosed. Therefore, making it more comprehensive about the mechanisms of BBMs is emergently necessary. Several studies have demonstrated that tumor microenvironment has potent roles in the tumor progression and metastasis. Besides, according to the former studies, cells in different organs using glucose might be various; as well as the dominant glucose transporters used are distinct. Here, we analyzed the brain metastatic breast cancer (BR) cells collected from intracardiac injection of nude mice and found the utilization of glucose transporters in the BR cells were different from the parental breast cancer cells; the elevated expressions of glucose transporter 3 were observed. In addition, the higher anaerobic glycolysis ability was found in BR cells. And after GLUT3 knockdown, the glucose reprogramming in BR cells was significantly decreased as well as the cancer malignant behaviors both in vitro and in vivo. Besides manipulating GLUT3 expressions, we also provide a potential mechanism that CREB could be a potent regulator of GLUT3 in BR cells by chromatin immunoprecipitation (ChIP) and knockdown assay. After decreasing the expressions of CREB in BR cells, the expressions of GLUT3 were also significantly reduced. In sum, our results reveal that GLUT3 plays an important role in provoking breast cancer cells surviving in brain by mediating glucose utilizations. And these results could provide a potential therapeutic way to target BBMs.Support or Funding Information1. Ministry of Science and Technology, R.O.C 2. Molecular Medicine Taiwan International Graduate Program, Academia Sinica
Anti‐cancer Effects of Melatonin, Estrogen, and Progesterone Hormone Therapy in MCF‐7 and MDA‐MB‐231 Breast Cancer Cells
Authors: Hasan Mahmud , Browne Erin , Guarinoni Laura , Darveau Travis , Hilton Katherine , Witt-Enderby Paula A. 
Published: April 2020

A novel melatonin, estrogen (E2), and progesterone (P4) hormone therapy (MEPT) was developed as a safe bio‐identical alternative hormone therapy for menopausal women based on the Women’s Health Initiative findings that PremPro™ increased breast cancer (BC) risk and mortality of all types of BC in postmenopausal women. MEPT delayed tumor onset and reduced tumor incidence in Neu female mice (Dodda et al., 2019; Front Oncol). For other BCs, its actions are currently unknown. In this study, MEPT’s anti‐cancer actions (cell viability, migration) were tested in MCF‐7 (human ER+/PR+) and MDA‐MB‐231 (human TNBC with constitutively active ras mutation) using equivalent concentrations of melatonin (1nM), E2 (50pM), and P4 (10nM) observed in serum after an oral dose that produced anti‐tumor actions in neu mice (Dodda et al 2019; Front Oncol). Potential mechanisms underlying MEPT’s anti‐cancer actions were assessed by western blot (pERK1/2, pERK5, NF‐kB, RUNX2, β1‐INTEGRIN, RANKL, and ELF‐5) using MEK1/2 (PD98059) and MEK5 (Bix02189) inhibitors and radioligand binding using 2‐[125I]‐iodomelatonin or [3H]estradiol binding for melatonin and estrogen receptor expression, respectively. For MCF‐7 cells, BIX02189 combined with MEPT blocked MEPT‐mediated inhibition of migration and MEPT‐mediated ELF5 induction in MCF‐7 cells (vs. vehicle) and decreased pERK5 (vs. Bix02189) suggesting that MEK5, through pERK1/2 and ELF5, were involved in the anti‐cancer actions of MEPT in MCF‐7 cells. For MDA‐MB‐231 cells, although PD98059 and Bix02189 alone inhibited viability and migration, when added in combination with MEPT, no further blockade or enhancement occurred suggesting that MEK1/2 and MEK5 were not involved in MEPT’s anti‐cancer actions in MDA‐MB‐231 cells. MEPT’s differential actions on MCF‐7 and MDA‐MB‐231 cells were most likely attributed to unique melatonin/melatonin receptor actions within each cell. Both MCF‐7 and MDA‐MB‐231 cells express high‐affinity melatonin receptors (MCF‐7: Kd = 13 pM; Bmax = 20.55 fmol/mg protein vs. MDA‐231: Kd = 1.7 nM; Bmax = 60.09 fmol/mg protein) but only in MCF‐7 cells were [3H]‐estradiol binding sites detected (MCF‐7: Kd = 2.2 nM; Bmax = 8.6 pmol/mg protein) and stimulatory actions of E2 or P4 on MCF‐7 cell proliferation observed. These findings demonstrate novel anti‐cancer actions of MEPT in ER+ BC and TNBC cells through distinct mechanisms. Future studies assessing MEPT’s actions against ER+ BC and TNBC in vivo are warranted.Support or Funding InformationThis study was funded by the Marie‐Clement Rodier, C.S.Sp. Endowed Chair.
Chronic Hypertension Increases Pial Perivascular Microglia in Female Dahl‐S Rats
Authors: Warrington Junie Paula , Shao Qingmei , Clayton Ahsia M. , Maeda Kenji J. , Garrett Michael R. , Sasser Jennifer M. 
Published: April 2020

Recent studies have shown that hypertension increases the proliferation and migration of microglia towards the perivascular space before destruction of the vasculature. Other studies have shown that hypertension increases perivascular macrophages, shown to contribute to impaired neurovascular coupling and cognitive impairments in mice. Despite these studies, there has been no description of what happens to perivascular microglia (PVMG) at the pia in the setting of chronic hypertension or following pregnancy.The goal of this study was to test the hypothesis that chronic hypertension increases the density of PVMGs and that complicated pregnancy history will exacerbate this increase.Female (8 months old) Sprague Dawley and Dahl‐S virgin or prior pregnant (2 pregnancies) rats were used. Brains were collected 3.5 months after the second pregnancy and processed for immunofluorescence staining for ion calcium binding adapter molecule (Iba‐1, a microglia marker) and DAPI. Z‐stacks were captured (0.5μm steps) throughout the thickness of the slice (20μm) using confocal microscopy. The number of microglia surrounding the pial artery (posterior communicating artery) was counted and normalized to the circumference of the vessel (density). We also measured the distance between each microglia surrounding the vessel.There was no effect of pregnancy on PVMG density (p=0.675) or mean distance between PVMG (p=0.161) but a significant effect of hypertension (p<0.001) on both measures. PVMG density increased from 0.017±0.005 to 0.031±0.004/μm in the virgins and from 0.020±0.002 to 0.032±0.002/μm in the prior pregnant rats. Mean distance between PVMGs decreased from 64±13 to 28±5μm in the virgins and from 52±7 to 21±2 μm in the prior pregnant rats. Some of the PVMGs were antigen presenting cells (co‐expressed major histocompatibility complex II, MHCII).Our results indicate that at the pia, PVMGs accumulate around the blood vessels, with some presenting antigens to lymphocytes. The exact function of these cells are not known, but we hypothesize that PVMGs could be contributing to the pathophysiology of hypertension or could present a compensatory mechanism to protect the blood vessels. Future studies will directly test the function of pial PVMGs in the context of acute and chronic hypertension and will assess whether this finding is specific to female brains.Support or Funding InformationUMMC Department of Neurology