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Colleen E Dugan, James P Grinias, Sebastian D Parlee, Mahmoud El-Azzouny, Charles R Evans, Robert T Kennedy
Microfluidics is an enabling technology for both cell biology and chemical analysis. We combine these attributes with a microfluidic device for on-line solid-phase extraction (SPE) and mass spectrometry (MS) analysis of secreted metabolites from living cells in culture on the chip. The device was constructed with polydimethylsiloxane (PDMS) and contains a reversibly sealed chamber for perfusing cells. A multilayer design allowed a series of valves to control an on-chip 7.5 μL injection loop downstream of the cell chamber with operation similar to a six-port valve...
October 19, 2016: Analytical and Bioanalytical Chemistry
Ru Zheng, Yue Li, Huiying Sun, Xiaoyin Lu, Bao-Fa Sun, Rui Wang, Lina Cui, Cheng Zhu, Hai-Yan Lin, Hongmei Wang
The syncytiotrophoblast (STB) plays a key role in maintaining the function of the placenta during human pregnancy. However, the molecular network that orchestrates STB development remains elusive. The aim of this study was to obtain broad and deep insight into human STB formation via transcriptomics. We adopted RNA sequencing (RNA-Seq) to investigate genes and isoforms involved in forskolin (FSK)-induced fusion of BeWo cells. BeWo cells were treated with 50 μM FSK or dimethylsulfoxide (DMSO) as a vehicle control for 24 and 48 h, and the mRNAs at 0, 24 and 48 h was sequenced...
October 14, 2016: Reproduction: the Official Journal of the Society for the Study of Fertility
Luigi Sapio, Monica Gallo, Michela Illiano, Emilio Chiosi, Daniele Naviglio, Annamaria Spina, Silvio Naviglio
Cancer is a major public health problem and the second leading cause of mortality around the world. Although continuous advances in the science of oncology and cancer research are now leading to improved outcomes for many cancer patients, novel cancer treatment options are strongly demanded. Naturally occurring compounds from a variety of vegetables, fruits and medicinal plants have been shown to exhibit various anticancer properties in a number of in vitro and in vivo studies and represent an attractive research area for the development of new therapeutic strategies to fight cancer...
October 14, 2016: Journal of Cellular Physiology
Rauf Latif, Ronald B Realubit, Charles Karan, Mihaly Mezei, Terry F Davies
Pathological activation of the thyroid-stimulating hormone receptor (TSHR) is caused by thyroid-stimulating antibodies in patients with Graves' disease (GD) or by somatic and rare genomic mutations that enhance constitutive activation of the receptor influencing both G protein and non-G protein signaling. Potential selective small molecule antagonists represent novel therapeutic compounds for abrogation of such abnormal TSHR signaling. In this study, we describe the identification and in vitro characterization of a novel small molecule antagonist by high-throughput screening (HTS)...
2016: Frontiers in Endocrinology
Zhijun Sun, Xing Wu, Weiping Li, Hui Peng, Xuhua Shen, Lu Ma, Huirong Liu, Hongwei Li
BACKGROUND: Diabetes-induced vascular dysfunction may arise from reduced nitric oxide (NO) availability, following interaction with superoxide to form peroxynitrite. Peroxynitrite can induce formation of 3-nitrotyrosine-modified proteins. RhoA/ROCK signaling is also involved in diabetes-induced vascular dysfunction. The study aimed to investigate possible links between Rho/ROCK signaling, hyperglycemia, and peroxynitrite in small coronary arteries. METHODS: Rat small coronary arteries were exposed to normal (NG; 5...
October 11, 2016: BMC Cardiovascular Disorders
Edgar Flores-Soto, Jorge Reyes-García, Abril Carbajal-García, Elías Campuzano-González, Mercedes Perusquía, Bettina Sommer, Luis M Montaño
: Testosterone (TES), other androgens and female sex steroids induce non-genomic rapid relaxing effects in airway smooth muscle (ASM). In guinea pig ASM, basal tension was relaxed by dehydroepiandrosterone (DHEA) and TES; 17β-estradiol (E2) had a small effect. Blockers of L-type voltage dependent Ca(2+) channel (L-VDCC, D-600) and store operated Ca(2+) channel (SOC, 2-APB) also relaxed the basal tone. In tracheal myocytes, DHEA and TES diminished intracellular basal Ca(2+) concentrations (b[Ca(2+)]i) as D-600+2-APB but to a higher extend...
October 4, 2016: Molecular and Cellular Endocrinology
Riyaad Aungraheeta, Alexandra Conibear, Mark Butler, Eamonn Kelly, Sven Nylander, Andrew Mumford, Stuart Mundell
Ticagrelor is a potent antagonist of the P2Y12 receptor (P2Y12R) and consequently an inhibitor of platelet activity effective in the treatment of atherothrombosis. Here, we sought to further characterise its molecular mechanism of action. Initial studies showed that ticagrelor promoted a greater inhibition of ADP-induced Ca(2+) release in washed platelets versus other P2Y12R antagonists. This additional effect of ticagrelor beyond P2Y12R antagonism was in part as a consequence of ticagrelor inhibiting the equilibrative nucleoside transporter 1 (ENT1) on platelets, leading to accumulation of extracellular adenosine and activation of Gs-coupled adenosine A2A receptors...
September 30, 2016: Blood
Tomoyuki Kaneko, Katsuyuki Kaneda, Atsushi Ohno, Daiki Takahashi, Taiki Hara, Taiju Amano, Soichiro Ide, Mitsuhiro Yoshioka, Masabumi Minami
Pain is a complex experience involving sensory and affective components. Although the neuronal mechanisms underlying the sensory component of pain have been extensively studied, those underlying its affective component have yet to be elucidated. Recently, we reported that corticotrophin-releasing factor (CRF)-induced depolarization in type II neurons within the dorsolateral bed nucleus of the stria terminalis (dlBNST) is critical for pain-induced aversive responses in rats. However, the intracellular signaling underlying the excitatory effects of CRF and the contribution of such signaling to the induction of pain-induced aversion remain unclear...
September 30, 2016: European Journal of Neuroscience
Angela Yulia, Natasha Singh, Kaiyu Lei, Suren R Sooranna, Mark R Johnson
The factors that initiate human labor are poorly understood. We have tested the hypothesis that a decline in cAMP/PKA function leads to the onset of labor. Initially, we identified myometrial cAMP/PKA responsive genes (6 up-regulated and 5 down-regulated genes) and assessed their expression in myometrial samples taken from different stages of pregnancy and labor. We found that the oxytocin receptor (OTR) was one of the cAMP repressed genes and, given the importance of OTR in the labor process, we studied the mechanisms involved in greater detail using siRNA, chemical agonists and antagonists of the cAMP effectors...
September 27, 2016: Endocrinology
Hong-Ping Yu, Nan Zhang, Ting Zhang, Zi-Long Wang, Ning Li, Hong-Hai Tang, Run Zhang, Meng-Na Zhang, Biao Xu, Quan Fang, Rui Wang
Neurite outgrowth is an important process in neural regeneration and plasticity, especially after neural injury, and recent evidence indicates that several Gαi/o protein-coupled receptors play an important role in neurite outgrowth. The neuropeptide (NP)FF system contains two Gαi/o protein-coupled receptors, NPFF1 and NPFF2 receptors, which are mainly distributed in the central nervous system. The aim of the present study was to determine whether the NPFF system is involved in neurite outgrowth in Neuro 2A cells...
September 23, 2016: Peptides
Minji Lee, Wonyoung Koh, Bomee Kim, Hyeju Chung, Gahyang Cho, Haekwon Kim
Previously we observed that human adipose-derived stem cells (hADSCs) could form aggregation during culture in the presence of human serum (HS). In the present study, we have examined if the aggregation might result from the cell migration and analyzed the difference of cell adhesivity after culture in various conditions. When cells were cultured in fetal bovine serum (FBS) alone, there was no morphological change. Similarly, cells pretreated with FBS for 1 day or cultured in a mixture of FBS and HS showed little change...
June 2016: Balsaeng'gwa Saengsig
Jingquan Sun, Xin Ye, Minhao Xie, Jianping Ye
Muscle exercise induces intramuscular triglyceride (TG) accumulation and promotes mitochondrial maintenance in myotubes. However, the mechanism underlying exercise effects remains unknown. In this study, lactic acid was tested as a signaling molecule in C2C12 myotubes to understand the mechanism. Intracellular TG storage was induced in the cells by sodium lactate. The lactate activity was observed with an inhibition of the cAMP-PKA pathway as indicated by a reduction in the phosphorylation status of CREB (pCREB)...
2016: Scientific Reports
Daniela Martins Paschoal, Mateus José Sudano, Kátia Regina Lancellotti Schwarz, Rosiára Rosário Dias Maziero, Midyan Daroz Guastali, Letícia Ferrari Crocomo, Luis Carlos Oña Magalhães, Alício Martins, Claudia Lima Verde Leal, Fernanda da Cruz Landim-Alvarenga
The presence of fetal calf serum in culture medium influences embryo quality, causing a reduction in postcryopreservation survival. Forskolin has been used to induce lipolysis and increase cryotolerance, functioning as an activator of adenylate cyclase and elevating cAMP levels. In the present experiment, bovine zygotes were cultured in synthetic oviduct fluid with amino acid plus 2.5% fetal calf serum for 6 days, when forskolin was added in three concentrations: 2.5, 5, and 10 μM. Treatment with forskolin lasted for 24 hours...
August 18, 2016: Theriogenology
Antonio Dore, Battistina Asproni, Alessia Scampuddu, Stefania Gessi, Gabriele Murineddu, Elena Cichero, Paola Fossa, Stefania Merighi, Serena Bencivenni, Gérard A Pinna
Pyrazolo[5,1-f][1,6]naphthyridine-carboxamide derivatives were synthesized and evaluated for the affinity at CB1 and CB2 receptors. Based on the AgOTf and proline-cocatalyzed multicomponent methodology, the ethyl 5-(p-tolyl)pyrazolo[5,1-f][1,6]naphthyridine-2-carboxylate (12) and ethyl 5-(2,4-dichlorophenyl)pyrazolo[5,1-f][1,6]naphthyridine-2-carboxylate (13) intermediates were synthesized from the appropriate o-alkynylaldehydes, p-toluenesulfonyl hydrazide and ethyl pyruvate. Most of the novel compounds feature a p-tolyl (8a-i) or a 2,4-dichlorophenyl (8j) motif at the C5-position of the tricyclic pyrazolo[5,1-f][1,6]naphthyridine scaffold...
November 1, 2016: Bioorganic & Medicinal Chemistry
Hai Jin, Guorong Wen, Shili Deng, Shuo Wan, Jingyu Xu, Xuemei Liu, Rui Xie, Hui Dong, Biguang Tuo
The incidence of duodenal ulcer is markedly lower in women than men, but the cause of the gender difference is not clear. The cystic fibrosis transmembrane conductance regulator (CFTR) and the solute linked carrier 26 gene family A6 (SLC26A6) are two key bicarbonate transport proteins to mediate duodenal mucosal bicarbonate secretion which is an important protective factor against acid-induced duodenal injury. This study aimed to investigate the effect of estrogen on the expressions and functional activities of CFTR and SLC26A6 in duodenal mucosa...
September 12, 2016: Experimental Physiology
Nermeen N El-Agroudy, Reem N El-Naga, Rania Abd El-Razeq, Ebtehal El-Demerdash
BACKGROUND AND PURPOSE: Liver fibrosis is one of the leading causes of morbidity and mortality worldwide with very limited therapeutic options. Given the pivotal role of activated hepatic stellate cells in liver fibrosis, attention has been directed towards the signalling pathways underlying their activation and fibrogenic functions. Recently, the hedgehog (Hh) signalling pathway has been identified as a potentially important therapeutic target in liver fibrosis. The present study was designed to explore the antifibrotic effects of the potent Hh signalling inhibitor, forskolin, and the possible molecular mechanisms underlying these effects...
September 2, 2016: British Journal of Pharmacology
Takayuki Matsumoto, Makoto Ando, Shun Watanabe, Maika Iguchi, Mako Nagata, Shota Kobayashi, Kumiko Taguchi, Tsuneo Kobayashi
In cellular events, endoplasmic reticulum (ER) stress has an important role in the development of various diseases including cardiovascular diseases. Tunicamycin, an inhibitor of N-linked glycosylation, is known to be an inducer of ER stress. However, the extent to which tunicamycin affects the vasorelaxant function is not completely understood. Thus, we investigated the effect of tunicamycin on relaxations induced by various vasorelaxant agents, including acetylcholine (ACh; endothelium-dependent vasodilator), sodium nitroprusside (SNP; endothelium-independent vasodilator), isoprenaline (ISO; beta-adrenoceptor agonist), forskolin (FSK; adenylyl cyclase activator), and cromakalim [ATP-sensitive K(+) (KATP) channel activator] in organ-cultured superior mesenteric arteries of rats, which are treated with either a vehicle [dimethyl sulfoxide (DMSO)] or tunicamycin (20 µg/mL for 22-24 h)...
2016: Biological & Pharmaceutical Bulletin
Brenna C Beckelman, Stephen Day, Xueyan Zhou, Maggie Donohue, Gunnar K Gouras, Eric Klann, C Dirk Keene, Tao Ma
Synaptic dysfunction may represent an early and crucial pathophysiology in Alzheimer's disease (AD). Recent studies implicate a connection between synaptic plasticity deficits and compromised capacity of de novo protein synthesis in AD. The mRNA translational factor eukaryotic elongation factor 1A (eEF1A) is critically involved in several forms of long-lasting synaptic plasticity. By examining postmortem human brain samples, a transgenic mouse model, and application of synthetic human Aβ42 on mouse hippocampal slices, we demonstrated that eEF1A protein levels were significantly decreased in AD, particularly in the hippocampus...
September 6, 2016: Journal of Alzheimer's Disease: JAD
Robbert Havekes, Alan J Park, Rosa E Tolentino, Vibeke M Bruinenberg, Jennifer C Tudor, Yool Lee, Rolf T Hansen, Leonardo A Guercio, Edward Linton, Susana R Neves-Zaph, Peter Meerlo, George S Baillie, Miles D Houslay, Ted Abel
UNLABELLED: Alterations in cAMP signaling are thought to contribute to neurocognitive and neuropsychiatric disorders. Members of the cAMP-specific phosphodiesterase 4 (PDE4) family, which contains >25 different isoforms, play a key role in determining spatial cAMP degradation so as to orchestrate compartmentalized cAMP signaling in cells. Each isoform binds to a different set of protein complexes through its unique N-terminal domain, thereby leading to targeted degradation of cAMP in specific intracellular compartments...
August 24, 2016: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Marina Ayres Pereira, Thomas Mandel Clausen, Caroline Pehrson, Yang Mao, Mafalda Resende, Mads Daugaard, Anders Riis Kristensen, Charlotte Spliid, Line Mathiesen, Lisbeth E Knudsen, Peter Damm, Thor G Theander, Stefan R Hansson, Morten A Nielsen, Ali Salanti
During placental malaria, Plasmodium falciparum infected erythrocytes sequester in the placenta, causing health problems for both the mother and fetus. The specific adherence is mediated by the VAR2CSA protein, which binds to placental chondroitin sulfate (CS) on chondroitin sulfate proteoglycans (CSPGs) in the placental syncytium. However, the identity of the CSPG core protein and the cellular impact of the interaction have remain elusive. In this study we identified the specific CSPG core protein to which the CS is attached, and characterized its exact placental location...
August 2016: PLoS Pathogens
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