bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mediator subunit OsMED14_1 plays an important role in rice development
Authors: Malik Naveen , Ranjan Rajeev , Parida Swarup K. , Agarwal Pinky , Tyagi Akhilesh K. 
Published: November 2019

SummaryMediator, a multisubunit co‐activator complex, regulates transcription in eukaryotes and is involved in diverse processes in Arabidopsis through its different subunits. Here, we have explored developmental aspects of one of the rice Mediator subunit gene OsMED14_1. We analyzed its expression pattern through RNA in situ hybridization and pOsMED14_1:GUS transgenics that showed its expression in roots, leaves, anthers and seeds prominently at younger stages, indicating possible involvement of this subunit in multiple aspects of rice development. To understand the developmental roles of OsMED14_1 in rice, we generated and studied RNAi‐based knockdown rice plants that showed multiple effects including less height, narrower leaves and culms with reduced vasculature, lesser lateral root branching, defective microspore development, reduced panicle branching and seed set, and smaller seeds. Histological analyses showed that slender organs were caused by reduction in both cell number and cell size in OsMED14_1 knockdown plants. Flow cytometric analyses and expression analyses of cell cycle‐related genes revealed that defective cell‐cycle progression led to these defects. Expression analyses of auxin‐related genes and indole‐3‐acetic acid (IAA) immunolocalization study indicated altered auxin level in these knockdown plants. Reduction of lateral root branching in knockdown plants was corrected by exogenous IAA supplement. OsMED14_1 physically interacts with transcription factors YABBY5, TAPETUM DEGENERATION RETARDATION (TDR) and MADS29, possibly regulating auxin homeostasis and ultimately leading to lateral organ/leaf, microspore and seed development.
Analytical performance of canine acylated and unacylated ghrelin enzyme‐linked immunosorbent assays
Authors: Lass Julie , Nielsen Mette L. , Bjørnvad Charlotte R. , Vitger Anne D. , Cremer Signe E. 
Published: November 2019

AbstractObjectivesGhrelin is a major appetite‐stimulating hormone. It circulates as acylated ghrelin (AG) and unacylated ghrelin (UAG), which could have different metabolic actions in obesity. Our objective was to study the analytical performance of two new canine AG and UAG ELISAs using blood samples from healthy, normal‐weight dogs. Additionally, the effect of a protease inhibitor (PI) on short‐term sample storage was analyzed.MethodsThe intra‐ and inter‐assay precision for low, intermediate, and high AG and UAG concentrations, accuracy, limits of quantification (LQ), and detection limit (DL) of a blank sample were determined in ten healthy dogs. To study the effects of a PI on ghrelin concentrations, and AG and UAG concentrations were compared in five canine plasma samples stored for 1 month with (PI+), without (PI−), and with PI added at sample thawing (PI+th).ResultsThe intra‐ and inter‐assay coefficients of variation were 1.8%‐5.7% and 2.9%‐6.4% for the AG assay, and 0.8%‐7.5% and 2.8%‐13.4% for the UAG assay, respectively. Accuracy analyses showed nonsignificant deviation from linearity for the AG (R2 = .99; Runs test: P = .37) and UAG (R2 = .99; Runs test: P = .42) assays. For the AG assay, the upper LQ was >1261 pg/mL, the lower LQ was 6.2 pg/mL, and the DL was 0.3 pg/mL. For the UAG assay, the upper LQ was >1785 pg/mL, the lower LQ was 16.3 pg/mL, and the DL was 1.8 pg/mL. No differences in the AG (P = .54) and UAG (P = .95) concentrations were detected in the plasma samples subjected to PI+, PI−, and PI+th.ConclusionThe AG and UAG ELISA assays had acceptable precision, accuracy, lower LQ, and DLs, but upper LQ could not be established. An influence of the PI on short‐term storage was not detectable. Long‐term storage ± PI was not evaluated and should be investigated further.
Insulin‐induced membrane permeability to glucose in human muscles at rest and following exercise
Authors: McConell Glenn K. , Sjøberg Kim A. , Ceutz Frederik , Gliemann Lasse , Nyberg Michael , Hellsten Ylva , Frøsig Christian , Kiens Bente , Wojtaszewski Jørgen F. P. , Richter Erik A. 
Published: November 2019

Key points
Increased insulin action is an important component of the health benefits of exercise, but its regulation is complex and not fully elucidated.
Previous studies of insulin‐stimulated GLUT4 translocation to the skeletal muscle membrane found insufficient increases to explain the increases in glucose uptake.
By determination of leg glucose uptake and interstitial muscle glucose concentration, insulin‐induced muscle membrane permeability to glucose was calculated 4 h after one‐legged knee‐extensor exercise during a submaximal euglycaemic–hyperinsulinaemic clamp.
It was found that during submaximal insulin stimulation, muscle membrane permeability to glucose in humans increases twice as much in previously exercised vs. rested muscle and outstrips the supply of glucose, which then becomes limiting for glucose uptake.
This methodology can now be employed to determine muscle membrane permeability to glucose in people with diabetes, who have reduced insulin action, and in principle can also be used to determine membrane permeability to other substrates or metabolites.
AbstractIncreased insulin action is an important component of the health benefits of exercise, but the regulation of insulin action in vivo is complex and not fully elucidated. Previously determined increases in skeletal muscle insulin‐stimulated GLUT4 translocation are inconsistent and mostly cannot explain the increases in insulin action in humans. Here we used leg glucose uptake (LGU) and interstitial muscle glucose concentration to calculate insulin‐induced muscle membrane permeability to glucose, a variable not previously possible to quantify in humans. Muscle membrane permeability to glucose, measured 4 h after one‐legged knee‐extensor exercise, increased ∼17‐fold during a submaximal euglycaemic–hyperinsulinaemic clamp in rested muscle (R) and ∼36‐fold in exercised muscle (EX). Femoral arterial infusion of NG‐monomethyl l‐arginine acetate or ATP decreased and increased, respectively, leg blood flow (LBF) in both legs but did not affect membrane glucose permeability. Decreasing LBF reduced interstitial glucose concentrations to ∼2 mM in the exercised but only to ∼3.5 mM in non‐exercised muscle and abrogated the augmented effect of insulin on LGU in the EX leg. Increasing LBF by ATP infusion increased LGU in both legs with uptake higher in the EX leg. We conclude that it is possible to measure functional muscle membrane permeability to glucose in humans and it increases twice as much in exercised vs. rested muscle during submaximal insulin stimulation. We also show that muscle perfusion is an important regulator of muscle glucose uptake when membrane permeability to glucose is high and we show that the capillary wall can be a significant barrier for glucose transport.
Effectiveness of a multilevel intervention to reduce violence and increase condom use in intimate partnerships among female sex workers: cluster randomised controlled trial in Karnataka, India
Authors: Javalkar Prakash , Platt Lucy , Prakash Ravi , Beattie Tara S , Collumbien Martine , Gafos Mitzy , Ramanaik Satyanarayana , Davey Calum , Jewkes Rachel , Watts Charlotte , Bhattacharjee Parinita , Thalinja Raghavendra , DL Kavitha , Isac Shajy , Heise Lori 
Published: November 2019

IntroductionSamvedana Plus is a multilevel intervention working with sex workers, their intimate partners (IPs) and communities to reduce intimate partner violence (IPV) and to increase condom use within intimate relationships of sex workers in Northern Karnataka, India.MethodsA cluster randomised controlled trial in 47 villages. Female sex workers with IPs in the last 6 months were eligible for baseline (2014), midline (2016) and endline (2017) surveys. 24 villages were randomised to Samvedana Plus and 23 to a wait-list control. Primary outcomes among sex workers included experience of physical and/or sexual IPV or severe physical/sexual IPV in the last 6 months and consistent condom use with their IP in past 30 days. Analyses adjusted for clustering and baseline cluster-level means of outcomes.ResultBaseline (n=620) imbalance was observed with respect to age (33.9 vs 35.2) and IPV (31.4% vs 45.0%). No differences in physical/sexual IPV (8.1% vs 9.0%), severe physical/sexual IPV (6.9% vs 8.7%) or consistent condom use with IPs (62.5% vs 57.3%) were observed by trial arm at end line (n=547). Samvedana Plus was associated with decreased acceptance of IPV (adjusted OR (AOR)=0.62, 95% CI 0.40 to 0.94, p=0.025), increased awareness of self-protection strategies (AOR=1.73, 95% CI=1.04–2.89, p=0.035) and solidarity of sex workers around issues of IPV (AOR=1.69, 95% CI=1.02–2.82, p=0.042). We observed an increase in IPV between baseline (25.9%) and midline (63.5%) among women in Samvedana Plus villages but lower in comparison villages (41.8%–44.3%) and a sharp decrease at end line in both arms (~8%).ConclusionWe found no evidence that Samvedana Plus reduced IPV or increased condom use, but it may impact acceptance of IPV, increase knowledge of self-protection strategies and increase sex worker solidarity. Inconsistencies in reported IPV undermined the ability of the trial to assess effectiveness.Trial registration numberNCT02807259.
Muted responses to Ag accumulation by plankton to chronic and pulse exposure to silver nanoparticles in a boreal lake
Authors: Norman Beth C. , Frost Paul C. , Blakelock Graham C. , Higgins Scott N. , Hoque Md Ehsanul , Vincent Jennifer L. , Cetinic Katarina , Xenopoulos Marguerite A. 
Published: November 2019

Silver nanoparticles (AgNPs) are an emerging class of contaminants with the potential to impact ecosystem structure and function. AgNPs are antimicrobial, suggesting that microbe-driven ecosystem functions may be particularly vulnerable to AgNP exposure. Predicting the environmental impacts of AgNPs requires in situ investigation of environmentally relevant dosing regimens over time scales that allow for ecosystem-level responses. We used 3000 L enclosures installed in a boreal lake to expose plankton communities to chronic and pulse AgNP dosing regimens with concentrations mimicking those recorded in natural waters. We compared temporal patterns of plankton responses, Ag accumulation, and ecosystem metabolism (i.e., daily ecosystem respiration, gross primary production, and net ecosystem production) for 6 weeks of chronic dosing and 3 weeks following a pulsed dose. Ag accumulated in microplankton and zooplankton, but carbon-specific Ag was nonlinear over time and generally did not predict plankton response. Ecosystem metabolism did not respond to either AgNP exposure type. This lack of response corresponded with weak microplankton responses in the chronic treatments but did not reflect the stronger microplankton response in the pulse treatment. Our results suggest that lake ecosystem metabolism is somewhat resistant to environmentally relevant concentrations of AgNPs and that organismal responses do not necessarily predict ecosystem-level responses.
Bimodal right ventricular dysfunction after postnatal hyperoxia exposure: implications for the preterm heart
Authors: Kumari Santosh , Braun Rudolf K. , Tetri Laura H. , Barton Gregory P. , Hacker Timothy A. , Goss Kara N. 
Published: November 2019

Rats exposed to postnatal hyperoxia develop right ventricular (RV) dysfunction, mild pulmonary hypertension, and dysregulated cardiac mitochondrial biogenesis when aged to one year, with the degree of cardiac dysfunction and pulmonary hypertension similar to that previously described in young adults born preterm. Here, we sought to understand the impact of postnatal hyperoxia exposure on RV hemodynamic and mitochondrial function across the life span. In Methods, pups from timed-pregnant Sprague-Dawley rats were randomized to normoxia or hyperoxia [fraction of inspired oxygen ([Formula: see text]), 0.85] exposure for the first 14 days of life, a commonly used model of chronic lung disease of prematurity. RV hemodynamic and mitochondrial function were assessed by invasive measurement of RV pressure-volume loops and by high-resolution respirometry at postnatal day 21 (P21), P90, and P365. In Results, at P21, hyperoxia-exposed rats demonstrated severe pulmonary hypertension and RV dysfunction, accompanied by depressed mitochondrial oxidative capacity. However, significant upregulation of mitochondrial biogenesis at P21 as well as improved afterload led to complete RV hemodynamic and mitochondrial recovery at P90. Mitochondrial DNA mutations were significantly higher by P90 and associated with significant late RV mitochondrial and hemodynamic dysfunction at P365. In conclusion, there appears to be a “honeymoon period” where cardiac hemodynamic and mitochondrial function normalizes following postnatal hyperoxia exposure, only to decline again with ongoing aging. This finding may have significant implications if a long-term pulmonary vascular screening program were to be developed for children or adults with a history of severe prematurity. Further investigation into the mechanisms of recovery are warranted.NEW & NOTEWORTHY Premature birth is associated with increased risk for cardiac dysfunction and failure throughout life. Here, we identify bimodal right ventricular dysfunction after postnatal hyperoxia exposure. Mitochondrial biogenesis serves as an early adaptive feature promoting recovery of cardiac hemodynamic and mitochondrial function. However, the accumulation of mitochondrial DNA mutations results in late mitochondrial and right ventricular dysfunction. This bimodal right ventricular dysfunction may have important implications for the development of screening programs in the preterm population.
AZD4573 Is a Highly Selective CDK9 Inhibitor That Suppresses MCL-1 and Induces Apoptosis in Hematologic Cancer Cells
Authors: Cidado Justin , Boiko Scott , Proia Theresa , Ferguson Douglas , Criscione Steven W. , San Martin Maryann , Pop-Damkov Petar , Su Nancy , Roamio Franklin Valar Nila , Sekhar Reddy Chilamakuri Chandra , D'Santos Clive S. , Shao Wenlin , Saeh Jamal C. , Koch Raphael , Weinstock David M. , Zinda Michael , Fawell Stephen E. , Drew Lisa 
Published: November 2019

Abstract

Purpose:
Cyclin-dependent kinase 9 (CDK9) is a transcriptional regulator and potential therapeutic target for many cancers. Multiple nonselective CDK9 inhibitors have progressed clinically but were limited by a narrow therapeutic window. This work describes a novel, potent, and highly selective CDK9 inhibitor, AZD4573.

Experimental Design:
The antitumor activity of AZD4573 was determined across broad cancer cell line panels in vitro as well as cell line- and patient-derived xenograft models in vivo. Multiple approaches, including integrated transcriptomic and proteomic analyses, loss-of-function pathway interrogation, and pharmacologic comparisons, were employed to further understand the major mechanism driving AZD4573 activity and to establish an exposure/effect relationship.

Results:
AZD4573 is a highly selective and potent CDK9 inhibitor. It demonstrated rapid induction of apoptosis and subsequent cell death broadly across hematologic cancer models in vitro, and MCL-1 depletion in a dose- and time-dependent manner was identified as a major mechanism through which AZD4573 induces cell death in tumor cells. This pharmacodynamic (PD) response was also observed in vivo, which led to regressions in both subcutaneous tumor xenografts and disseminated models at tolerated doses both as monotherapy or in combination with venetoclax. This understanding of the mechanism, exposure, and antitumor activity of AZD4573 facilitated development of a robust pharmacokinetic/PD/efficacy model used to inform the clinical trial design.

Conclusions:
Selective targeting of CDK9 enables the indirect inhibition of MCL-1, providing a therapeutic option for MCL-1–dependent diseases. Accordingly, AZD4573 is currently being evaluated in a phase I clinical trial for patients with hematologic malignancies (clinicaltrials.gov identifier: NCT03263637).
See related commentary by Alcon et al., p. 761
Genomic Determinants of Clinical Outcomes in Rhabdomyosarcoma
Authors: Casey Dana L. , Wexler Leonard H. , Pitter Kenneth L. , Samstein Robert M. , Slotkin Emily K. , Wolden Suzanne L. 
Published: November 2019

Abstract

Purpose:
Increased availability of next-generation sequencing has allowed for the genomic characterization of a variety of pediatric tumors, although genomic determinants of response to treatment remain largely unknown. We sought to evaluate the genomic landscape and genomic determinants of clinical outcomes in rhabdomyosarcoma (RMS).

Experimental Design:
Of 29,067 patients who underwent genomic profiling at our institution using a 468-gene oncopanel with complete records, 87 had RMS, of whom 22 were fusion positive. The 10 most common genetic alterations were associated with locoregional control (LC), disease-free survival (DFS), and overall survival (OS). Tumor mutational burden (TMB), defined as the total number of somatic nonsynonymous mutations normalized to the number of sequenced megabases, was also associated with clinical outcomes.

Results:
Median age at diagnosis was 16.4 years and median follow-up, 2.1 years. Patients with fusion-negative RMS had more genomic alterations and a higher TMB than those with fusion-positive RMS (mean number of genomic alterations, 6.0 vs. 2.9; P = 0.007 and mean TMB, 2.6 vs. 1.0; P = 0.01). Genetic alterations in TP53 were associated with worse OS (P = 0.03). High TMB (defined as the top quartile ≥ 2.8) was associated with worse LC (P = 0.05), DFS (P = 0.04), and OS (P = 0.01), with significance retained on multivariable analysis after controlling for risk group, fusion status, and receipt of chemotherapy as per pediatric protocols.

Conclusions:
High TMB was associated with worse clinical outcomes in patients with RMS. With further validation, TMB and other genomic classifiers may be combined with traditional clinicopathologic risk factors to guide risk stratification and ultimately treatment decisions.
Glucose 6-Phosphate Accumulates via Phosphoglucose Isomerase Inhibition in Heart Muscle
Authors: Karlstaedt Anja , Khanna Radhika , Thangam Manoj , Taegtmeyer Heinrich 
Published: November 2019

Rationale:
Metabolic and structural remodeling is a hallmark of heart failure. This remodeling involves activation of the mTOR (mammalian target of rapamycin) signaling pathway, but little is known on how intermediary metabolites are integrated as metabolic signals.

Objective:
We investigated the metabolic control of cardiac glycolysis and explored the potential of glucose 6-phosphate (G6P) to regulate glycolytic flux and mTOR activation.

Methods and Results:

We developed a kinetic model of cardiomyocyte carbohydrate metabolism,
CardioGlyco
, to study the metabolic control of myocardial glycolysis and G6P levels. Metabolic control analysis revealed that G6P concentration is dependent on phosphoglucose isomerase (PGI) activity. Next, we integrated ex vivo tracer studies with mathematical simulations to test how changes in glucose supply and glycolytic flux affect mTOR activation. Nutrient deprivation promoted a tight coupling between glucose uptake and oxidation, G6P reduction, and increased protein-protein interaction between hexokinase II and mTOR. We validated the in silico modeling in cultured adult mouse ventricular cardiomyocytes by modulating PGI activity using erythrose 4-phosphate. Inhibition of glycolytic flux at the level of PGI caused G6P accumulation, which correlated with increased mTOR activation. Using click chemistry, we labeled newly synthesized proteins and confirmed that inhibition of PGI increases protein synthesis.

Conclusions:
The reduction of PGI activity directly affects myocyte growth by regulating mTOR activation.
Newly Identified Peptide, Peptide Lv, Promotes Pathological Angiogenesis
Authors: Shi Liheng , Zhao Min , Abbey Colette A. , Tsai Shu‐Huai , Xie Wankun , Pham Dylan , Chapman Samantha , Bayless Kayla J. , Hein Travis W. , Rosa Robert H. , Ko Michael L. , Kuo Lih , Ko Gladys Y.‐P. 
Published: November 2019

Background
We recently discovered a small endogenous peptide, peptide Lv, with the ability to activate vascular endothelial growth factor receptor 2 and its downstream signaling. As vascular endothelial growth factor through vascular endothelial growth factor receptor 2 contributes to normal development, vasodilation, angiogenesis, and pathogenesis of various diseases, we investigated the role of peptide Lv in vasodilation and developmental and pathological angiogenesis in this study.

Methods and Results
The endothelial cell proliferation, migration, and 3‐dimensional sprouting assays were used to test the abilities of peptide Lv in angiogenesis in vitro. The chick chorioallantoic membranes and early postnatal mice were used to examine its impact on developmental angiogenesis. The oxygen‐induced retinopathy and laser‐induced choroidal neovascularization mouse models were used for in vivo pathological angiogenesis. The isolated porcine retinal and coronary arterioles were used for vasodilation assays. Peptide Lv elicited angiogenesis in vitro and in vivo. Peptide Lv and vascular endothelial growth factor acted synergistically in promoting endothelial cell proliferation. Peptide Lv–elicited vasodilation was not completely dependent on nitric oxide, indicating that peptide Lv had vascular endothelial growth factor receptor 2/nitric oxide–independent targets. An antibody against peptide Lv, anti‐Lv, dampened vascular endothelial growth factor–elicited endothelial proliferation and laser‐induced vascular leakage and choroidal neovascularization. While the pathological angiogenesis in mouse eyes with oxygen‐induced retinopathy was enhanced by exogenous peptide Lv, anti‐Lv dampened this process. Furthermore, deletion of peptide Lv in mice significantly decreased pathological neovascularization compared with their wild‐type littermates.

Conclusions
These results demonstrate that peptide Lv plays a significant role in pathological angiogenesis but may be less critical during development. Peptide Lv is involved in pathological angiogenesis through vascular endothelial growth factor receptor 2–dependent and –independent pathways. As anti‐Lv dampened the pathological angiogenesis in the eye, anti‐Lv may have a therapeutic potential to treat pathological angiogenesis.