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https://www.readbyqxmd.com/read/28205171/synthesis-and-live-cell-imaging-of-fluorescent-sterols-for-analysis-of-intracellular-cholesterol-transport
#1
Maciej Modzel, Frederik W Lund, Daniel Wüstner
Cellular cholesterol homeostasis relies on precise control of the sterol content of organelle membranes. Obtaining insight into cholesterol trafficking pathways and kinetics by live-cell imaging relies on two conditions. First, one needs to develop suitable analogs that resemble cholesterol as closely as possible with respect to their biophysical and biochemical properties. Second, the cholesterol analogs should have good fluorescence properties. This interferes, however, often with the first requirement, such that the imaging instrumentation must be optimized to collect photons from suboptimal fluorophores, but good cholesterol mimics, such as the intrinsically fluorescent sterols, cholestatrienol (CTL) or dehydroergosterol (DHE)...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28203527/superoxide-increases-angiotensin-ii-at1-receptor-function-in-human-kidney-2-cells
#2
Mohammad Saleem, Indira Pokkunuri, Mohammad Asghar
The redox-sensitive Sp family transcription factor has been linked to the regulation of angiotensin II type 1 receptor (AT1R). However, the exact mechanism of AT1R regulation in renal cells is poorly understood. We tested the specificity of reactive oxygen species (ROS), superoxide vs. hydrogen peroxide (H2O2), and the specific role of Sp3 transcription factor, if any, in the regulation of AT1R in human kidney cells (HK2 cells). Superoxide dismutase (SOD) inhibitor diethyldithiocarbamate (DETC), but not H2O2 treatment, increased fluorescence levels of superoxide probe dihydroethidium (DHE)...
December 2016: FEBS Open Bio
https://www.readbyqxmd.com/read/28193034/leveraging-gas-phase-fragmentation-pathways-for-improved-identification-and-selective-detection-of-targets-modified-by-covalent-probes
#3
Scott B Ficarro, Christopher M Browne, Joseph D Card, William M Alexander, Tinghu Zhang, Eunyoung Park, Randall McNally, Sirano Dhe-Paganon, Hyuk-Soo Seo, Ilaria Lamberto, Michael J Eck, Sara J Buhrlage, Nathanael S Gray, Jarrod A Marto
The recent approval of covalent inhibitors for multiple clinical indications has reignited enthusiasm for this class of drugs. As interest in covalent drugs has increased, so too has the need for analytical platforms that can leverage their mechanism-of-action to characterize modified protein targets. Here we describe novel gas phase dissociation pathways which yield predictable fragment ions during MS/MS of inhibitor-modified peptides. We find that these dissociation pathways are common to numerous cysteine-directed probes as well as the covalent drugs, Ibrutinib and Neratinib...
December 20, 2016: Analytical Chemistry
https://www.readbyqxmd.com/read/28192190/%C3%AE-7-integrin-and-madcam-1-play-opposing-roles-during-the-development-of-non-alcoholic-steatohepatitis
#4
Hannah K Drescher, Angela Schippers, Thomas Clahsen, Hacer Sahin, Heidi Noels, Mathias Hornef, Norbert Wagner, Christian Trautwein, Konrad L Streetz, Daniela C Kroy
BACKGROUND & AIMS: Non-alcoholic steatohepatitis (NASH) is a leading cause of chronic liver disease in western countries. It is unclear how infiltrating leukocytes affect NASH-development. Hence our study aims to investigate the role of the homing/receptor-pair mucosal addressin cell adhesion molecule-1 (MAdCAM-1)/β7-Integrin for immune cell recruitment and disease progression in a steatohepatitis model. METHODS: Constitutive β7-Integrin deficient (β7(-/-)) and MAdCAM-1 deficient (MAdCAM-1(-/-)) mice were fed high-fat-diet (HFD) for 26 weeks or methionine-choline-deficient-diet (MCD) for 4 weeks...
February 9, 2017: Journal of Hepatology
https://www.readbyqxmd.com/read/28159361/enhanced-autophagy-by-exenatide-mitigates-doxorubicin-induced-cardiotoxicity
#5
Kyung Hye Lee, Haneul Cho, Sora Lee, Jong Shin Woo, Byung Hyun Cho, Jung Hee Kang, Yun-Mi Jeong, Xian Wu Cheng, Weon Kim
OBJECTIVES: Exenatide is a glucagon-like peptide-1 analogue that mitigates myocardial injury caused by ischemia-reperfusion injury via the survival signaling pathway. We hypothesized that exenatide would provide a protective effect in doxorubicin-induced cardiotoxicity. METHODS: H9c2 cardiomyocytes were pre-treated with exenatide followed by doxorubicin (DOX), and cell viability and intracellular reactive oxygen species (ROS) were subsequently measured. In order to determine the role of autophagy, we performed western blot as well as TUNEL and autophagosome staining...
January 27, 2017: International Journal of Cardiology
https://www.readbyqxmd.com/read/28116721/measurement-of-superoxide-production-and-nadph-oxidase-activity-by-hplc-analysis-of-dihydroethidium-oxidation
#6
Denise C Fernandes, Renata C Gonçalves, Francisco R M Laurindo
The fluorogenic probe dihydroethidium (DHE) is widely used for detecting intracellular superoxide. DHE oxidation by superoxide generates specifically the compound 2-hydroxyethidium (2-E(+)OH), so that 2-E(+)OH detection confers specificity to superoxide assessment among many other reactive oxygen species. However, DHE oxidation in biological systems leads to formation of other fluorescent products, particularly ethidium, usually formed at higher quantities than 2-E(+)OH. Since both 2-E(+)OH and ethidium are fluorescent, their identification and quantification is possible only after their physical separation by HPLC...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28115273/ablation-of-akt2-prevents-paraquat-induced-myocardial-mitochondrial-injury-and-contractile-dysfunction-role-of-nrf2
#7
Shuyi Wang, Xiaoling Zhu, Lize Xiong, Jun Ren
Paraquat is a quaternary nitrogen herbicide triggering oxidative stress, mitochondrial damage and multi-organ injuries including hearts. To date, effective measure to combat paraquat toxicity is still lacking. Recent evidence has revealed a role for Akt in cardiac homeostasis. To this end, this study was designed to examine the role of Akt2 in acute paraquat exposure-induced cardiac contractile and mitochondrial injury using a unique murine model of Akt2 knockout. Cardiac contractile and intracellular Ca(2+) properties were evaluated...
January 20, 2017: Toxicology Letters
https://www.readbyqxmd.com/read/27997907/improvement-of-huangqi-decoction-on-endothelial-dysfunction-in-5-6-nephrectomized-rats
#8
Shuang Chu, Li Wang, Xiao-Dong Mao, Wen Peng
BACKGROUND/AIMS: Endothelial dysfunction is a major factor in the progression of chronic kidney disease, which correlates with oxidative stress and NO deficiency. Huangqi decoction (HQD) is a potential anti-oxidant ingredient in renoprotection. However, the underlying mechanisms remained identified. Therefore, we investigated whether HQD exhibit improvement in endothelial dysfunction in the 5/6 nephrectomy (Nx) rat model. METHODS: Male Wistar rats (180 - 250 g) were divided into sham, Nx and Nx + HQD (0...
2016: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/27996306/mapping-qtls-associated-with-agronomic-and-physiological-traits-under-terminal-drought-and-heat-stress-conditions-in-wheat-triticum-aestivum-l
#9
Sirous Tahmasebi, Bahram Heidari, Hassan Pakniyat, C Lynne McIntyre
Wheat crops frequently experience a combination of abiotic stresses in the field, but most quantitative trait loci (QTL) studies have focused on the identification of QTLs for traits under single stress field conditions. A recombinant inbred line (RIL) population derived from SeriM82 × Babax was used to map QTLs under well-irrigated, heat, drought, and a combination of heat and drought stress conditions in two years. A total of 477 DNA markers were used to construct linkage groups that covered 1619.6 cM of the genome, with an average distance of 3...
January 2017: Genome Génome / Conseil National de Recherches Canada
https://www.readbyqxmd.com/read/27995330/piceatannol-attenuates-homocysteine-induced-endoplasmic-reticulum-stress-and-endothelial-cell-damage-via-heme-oxygenase-1-expression
#10
Jin-Sang Kil, Sun-Oh Jeong, Hun-Taeg Chung, Hyun-Ock Pae
A growing body of evidence implicates endoplasmic reticulum (ER)-induced cellular dysfunction and apoptosis as important factors to a variety of diseases. In endothelial cells (ECs), the sulfur-containing amino acid homocysteine (Hcy) causes EC apoptosis and reactive oxygen species (ROS) generation through induction of ER stress. Here, we have investigated whether piceatannol (Pic), a resveratrol analog, could protect ECs against Hcy-induced apoptosis, oxidative stress and ER stress, with specific emphasis on heme oxygenase-1 (HO-1)...
December 20, 2016: Amino Acids
https://www.readbyqxmd.com/read/27992310/a-kinetic-study-of-reactive-oxygen-species-in-rainbow-trout-hepatocytes-by-fluorometry
#11
Mazyar Yazdani, Ketil Hylland
The kinetics of reactive oxygen species (ROS) formation in a primary culture of rainbow trout hepatocytes was investigated using three fluorescent probes: 5-,6-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H2DCFDA), dihydrorhodamine 123 (DHR 123), and dihydroethidium (DHE). The cell cultures were loaded with the three probes, separately. Hepatocytes were then exposed to Cu (0.15-10 mM) in serum-free Leibovitz's medium for 30 min before being quantified by a fluorescence plate reader during 30 min...
2016: Journal of Environmental Pathology, Toxicology and Oncology
https://www.readbyqxmd.com/read/27893783/role-of-pten-in-oxidative-stress-and-dna-damage-in-the-liver-of-whole-body-pten-haplodeficient-mice
#12
Ezgi Eyluel Bankoglu, Oliver Tschopp, Johannes Schmitt, Philipp Burkard, Daniel Jahn, Andreas Geier, Helga Stopper
Type 2 diabetes (T2DM) and obesity are frequently associated with non-alcoholic fatty liver disease (NAFLD) and with an elevated cancer incidence. The molecular mechanisms of carcinogenesis in this context are only partially understood. High blood insulin levels are typical in early T2DM and excessive insulin can cause elevated reactive oxygen species (ROS) production and genomic instability. ROS are important for various cellular functions in signaling and host defense. However, elevated ROS formation is thought to be involved in cancer induction...
2016: PloS One
https://www.readbyqxmd.com/read/27882226/effect-of-dehydration-heat-exposure-on-thoracic-aorta-reactivity-in-rats
#13
Yao Geng, Lingqin Zhu, Fadong Liu, Xiaodan Zhu, Jianguo Niu, Guanghua Li
The aim of the present study was to investigate the effect of one week dehydration heat exposure on thoracic aorta reactivity in rats. Eighteen Male Sprague-Dawley rats were randomly divided into 3 groups (n=6 each group): Control group (CN), heat exposure group (HE), dehydration heat exposure group (DHE). The CN group was exposed to a room temperature of 24°C, while the HE and DHE groups were exposed to a heat temperature of 32°C. After 7 days of heat exposure, the heart rate and blood pressure of the rats were measured, and the noradrenaline (NA)-induced contraction on the aorta rings was measured by tension recording...
November 2016: Biomedical Reports
https://www.readbyqxmd.com/read/27863990/cytochrome-p450-enzymes-but-not-nadph-oxidases-are-the-source-of-the-nadph-dependent-lucigenin-chemiluminescence-in-membrane-assays
#14
Flávia Rezende, Kim-Kristin Prior, Oliver Löwe, Ilka Wittig, Valentina Strecker, Franziska Moll, Valeska Helfinger, Frank Schnütgen, Nina Kurrle, Frank Wempe, Maria Walter, Sven Zukunft, Bert Luck, Ingrid Fleming, Norbert Weissmann, Ralf P Brandes, Katrin Schröder
: Measuring NADPH oxidase (Nox)-derived reactive oxygen species (ROS) in living tissues and cells is a constant challenge. All probes available display limitations regarding sensitivity, specificity or demand highly specialized detection techniques. In search for a presumably easy, versatile, sensitive and specific technique, numerous studies have used NADPH-stimulated assays in membrane fractions which have been suggested to reflect Nox activity. However, we previously found an unaltered activity with these assays in triple Nox knockout mouse (Nox1-Nox2-Nox4(-/-)) tissue and cells compared to wild type...
January 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/27837002/safety-of-domperidone-in-treating-nausea-associated-with-dihydroergotamine-infusion-and-headache
#15
Nathaniel M Robbins, Hiroyuki Ito, Melvin M Scheinman, Peter J Goadsby
OBJECTIVE: To determine the safety of domperidone in the treatment of nausea associated with dihydroergotamine (DHE) infusion and headache. METHODS: We audited our use of domperidone for the inpatient management of nausea, focusing on known safety concerns, particularly potential cardiac arrhythmias. RESULTS: We reviewed 103 consecutive admissions of 90 patients admitted for IV DHE by infusion. Most admissions were to treat chronic migraine with (n = 53) or without (n = 46) aura...
December 13, 2016: Neurology
https://www.readbyqxmd.com/read/27816504/nadph-oxidase-nox-1-mediates-cigarette-smoke-induced-superoxide-generation-in-rat-vascular-smooth-muscle-cells
#16
Kyung-Hwa Chang, Jung-Min Park, Chang Hoon Lee, Bumseok Kim, Kyung-Chul Choi, Seong-Jin Choi, Kyuhong Lee, Moo-Yeol Lee
Smoking is a well-established risk factor for cardiovascular diseases. Oxidative stress is one of the common etiological factors, and NADPH oxidase (NOX) has been suggested as a potential mediator of oxidative stress. In this study, cigarette smoke (CS)-induced superoxide production was characterized in vascular smooth muscle cells (VSMC). CS was prepared in forms of cigarette smoke extract (CSE) and total particulate matter (TPM). Several molecular probes for reactive oxygen species were trialed, and dihydroethidium (DHE) and WST-1 were chosen for superoxide detection considering the autofluorescence, light absorbance, and peroxidase inhibitory activity of CS...
February 2017: Toxicology in Vitro: An International Journal Published in Association with BIBRA
https://www.readbyqxmd.com/read/27809355/aliskiren-protects-against-myocardial-ischemia-reperfusion-injury-via-an-endothelial-nitric-oxide-synthase-dependent-manner
#17
Yu Chen, Guoliang Meng, Wenli Bai, Yan Ma, Liping Xie, Naila Altaf, Yanning Qian, Yi Han, Yong Ji
Aliskiren, a direct renin inhibitor, was shown potent ability to attenuate hypertension. Our previous research has found that aliskiren protected against myocardial ischemia-reperfusion (I/R) injury and enhanced phosphorylation of endothelial nitric oxide synthase (eNOS) in spontaneously hypertensive rats. However, whether the cardioprotective effect of aliskiren against myocardial I/R injury was eNOS-dependent is unknown. In the present study, 12-week-old male eNOS knockout (eNOS(-/-) ) and wild-type C57BL/6J mice (WT) were orally administrated with the dose of 50 mg/kg/day of aliskiren...
November 3, 2016: Clinical and Experimental Pharmacology & Physiology
https://www.readbyqxmd.com/read/27804267/reactive-oxygen-species-and-oxidative-stress-in-obesity-recent-findings-and-empirical-approaches
#18
REVIEW
Fiona McMurray, David A Patten, Mary-Ellen Harper
OBJECTIVE: High levels of reactive oxygen species (ROS) are intricately linked to obesity and associated pathologies, notably insulin resistance and type 2 diabetes. However, ROS are also thought to be important in intracellular signaling, which may paradoxically be required for insulin sensitivity. Many theories have been developed to explain this apparent paradox, which have broadened our understanding of these important small molecules. While many sites for intracellular ROS production have been described, mitochondrial generated ROS remain a major contributor in most cell types...
November 2016: Obesity
https://www.readbyqxmd.com/read/27778035/-aliskiren-inhibits-proliferation-of-cardiac-fibroblasts-in-agt-ren-double-transgenic-hypertensive-mice-in-vitro
#19
Li-Ping Wang, Su-Jing Fan, Shu-Min Li, Xiao-Jun Wang, Na Sun
The purpose of the present study is to explore the effect of aliskiren on the proliferation of cardiac fibroblasts (CFs) in AGT-REN double transgenic hypertensive (dTH) mice. The cultured CFs from AGT-REN dTH mice were divided into AGT-REN group (dTH) and aliskiren group (ALIS). Cultured CFs from C57B6 mice were served as control (WT). The effect of different concentration of aliskiren (1 × 10(-6), 1 × 10(-7), 1 × 10(-8), 1 × 10(-9) mol/L) on CFs proliferation was determined by MTT assay. After treatment with 1 × 10(-7) mol/L aliskiren for 24 h, α-SMA, collagen I, III and NADPH oxidase (NOX) protein expression in CFs of AGT-REN dTH mice were detected by Western blot...
October 25, 2016: Sheng Li Xue Bao: [Acta Physiologica Sinica]
https://www.readbyqxmd.com/read/27775715/design-and-characterization-of-bivalent-bet-inhibitors
#20
Minoru Tanaka, Justin M Roberts, Hyuk-Soo Seo, Amanda Souza, Joshiawa Paulk, Thomas G Scott, Stephen L DeAngelo, Sirano Dhe-Paganon, James E Bradner
Cellular signaling is often propagated by multivalent interactions. Multivalency creates avidity, allowing stable biophysical recognition. Multivalency is an attractive strategy for achieving potent binding to protein targets, as the affinity of bivalent ligands is often greater than the sum of monovalent affinities. The bromodomain and extraterminal domain (BET) family of transcriptional coactivators features tandem bromodomains through which BET proteins bind acetylated histones and transcription factors...
December 2016: Nature Chemical Biology
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