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Find video protocols related to scientific articles indexed in Pubmed.
Communication: Ultrafast time-resolved ion photofragmentation spectroscopy of photoionization-induced proton transfer in phenol-ammonia complex.
J Chem Phys
PUBLISHED: 11-10-2014
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Photoionization-induced proton transfer (PT) in phenol-ammonia (PhOH-NH3) complex has been studied using ultrafast time-resolved ion photofragmentation spectroscopy. Neutral PhOH-NH3 complexes prepared in a free jet are photoionized by femtosecond [1+1] resonance-enhanced multiphoton ionization via the S1 state, and the subsequent dynamics occurring in the cations is probed by delayed pulses that result in ion fragmentation. The observed temporal evolutions of the photofragmentation spectra are consistent with an intracomplex PT reaction. The experiments revealed that PT in [PhOH-NH3](+) cation proceeds in two distinct steps: an initial impulsive wave-packet motion in ?70 fs followed by a slower relaxation of about 1 ps that stabilizes the system into the final PT configuration. These results indicate that for a barrierless PT system, even though the initial PT motions are impulsive and ultrafast, the reaction may take a much longer time scale to complete.
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Vibration Signals of Snoring as a Simple Severity Predictor for Obstructive Sleep Apnea.
Clin Respir J
PUBLISHED: 10-27-2014
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Polysomnography (PSG), which involves simultaneous monitoring of various physiological monitors, is the current comprehensive tool for diagnosing obstructive sleep apnea (OSA).
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Functional Characterization of Intracellular pH Regulators Responsible for Acid Extrusion in Human Radial Artery Smooth Muscle Cells.
Chin J Physiol
PUBLISHED: 09-23-2014
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Intracellular pH (pHi) is a critical factor influencing many important cellular functions. Acid extrusion carriers such as an Na?/H? exchanger (NHE) Na?/HCO? ? cotransporter (NBC) and monocarboxylate transporters (MCT) can be activated when cells are in an acidic condition (pHi < 7.1). Human radial artery smooth muscle cells (HRASMC) is an important conduit in coronary artery bypass graft surgery. However, such far, the pHi regulators have not been characterized in HRASMCs. We therefore investigated the mechanism of pHi recovery from intracellular acidosis and alkalosis, induced by NH?Cl-prepulse and Na-acetate-prepulse, respectively, using intracellular 2',7'-bis(2-carboxethyl)-5(6)- carboxy-fluorescein (BCECF)-fluorescence in HRASMCs. Cultured HRASMCs were derived from the segments of human radial artery that were obtained from patients undergoing bypass grafting. The resting pHi is 7.22 ± 0.03 and 7.17 ± 0.02 for HEPES- (nominally HCO? ?-free) and CO?/HCO??- buffered solution, respectively. In HEPES-buffered solution, a pHi recovery from induced intracellular acidosis could be blocked completely by 30 ?M HOE 694 (3-methylsulfonyl-4-piperidinobenzoyl, guanidine hydrochloride) a specific NHE inhibitor, or by removing [Na?]?. In 3% CO?/HCO? ?-buffered solution, HOE 694 slowed the pHi recovery from the induced intracellular acidosis only, while adding together with DIDS (a specific NBC inhibitor) or removal of [Na?]? entirely inhibited the acid extrusion. Moreover, ?-cyano-4-hydroxycinnamate (CHC; a specific blocker of MCT) blocked the lactate-induced pHi changes. In conclusion, we demonstrate, for the first time, that 3 different pHi regulators responsible for acid extruding, i.e. NHE and NBC, and MCT, are functionally co-existed in cultured HRASMCs.
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The effects of anterior vacuum disc on surgical outcomes of degenerative versus spondylolytic spondylolisthesis: at a minimum two-year follow-up.
BMC Musculoskelet Disord
PUBLISHED: 09-22-2014
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The vacuum phenomenon within the intervertebral disc usually represents disc degeneration. There are no reports in the English literature that focus on the effect of an anterior vacuum disc on surgical outcome of same-segment spondylolisthesis.
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In Situ Re-endothelialization via Multifunctional Nanoscaffolds.
ACS Nano
PUBLISHED: 09-15-2014
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The endothelium monolayer lining in the luminal side of blood vessels provides critical antithrombotic functions. Damage to these cells will expose a highly thrombogenic subendothelium, which leads to pathological vascular changes. Using combined tissue engineering and ligand-receptor targeting strategy, we developed a biodegradable urethane-doped polyester (UPE) multifunctional targeting nanoparticle (MTN) scaffold system with dual ligands: (1) glycoprotein 1b (GP1b) to target the injured arterial endothelium and subendothelium and (2) anti-CD34 antibodies to capture endothelial progenitor cells for endothelium regeneration. The fabricated spherical MTNs of 400 nm were found to be cytocompatible and hemocompatible. Both the in vitro and ex vivo targeting of these nanoscaffolds not only showed binding specificity of MTNs onto the von Willebrand factor -coated surfaces that simulate the injured arterial walls but also competed with platelets for binding onto these injured sites. Further in vivo study has revealed that a single delivery of MTNs upon vascular injury reduced neointimal hyperplasia by 57% while increased endothelium regeneration by ?60% in 21 days. These results support the promise of using MTN nanoscaffolds for treating vascular injury in situ.
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Transforaminal lumbar interbody debridement and fusion for the treatment of infective spondylodiscitis in the lumbar spine.
Eur Spine J
PUBLISHED: 09-12-2014
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To determine the safety and efficacy of using a single posterior approach with transforaminal lumbar interbody debridement and fusion (TLIDF) plus pedicle screws fixation in treating infective spondylodiscitis in the lumbar spine.
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TD-19, an erlotinib derivative, induces epidermal growth factor receptor wild-type nonsmall-cell lung cancer apoptosis through CIP2A-mediated pathway.
J. Pharmacol. Exp. Ther.
PUBLISHED: 09-03-2014
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Some patients with nonsmall-cell lung cancer (NSCLC) without epidermal growth factor receptor (EGFR) mutations still respond to gefitinib and erlotinib, suggesting that there may be a mechanism(s) other than the EGFR pathway that mediates the tumoricidal effects. In the current study, we tested the efficacy of TD-19, a novel compound chemically modified from erlotinib, which has more potent apoptotic effects than erlotinib in EGFR wild-type NSCLC cell lines. TD-19 induced significant cell death and apoptosis in H358, H441, H460, and A549 cells, as evidenced by increased caspase-3 activity and cleavage of procaspase-9 and poly (ADP-ribose) polymerase. The apoptotic effect of TD-19 in H460 cells, which were resistant to erlotinib, was associated with downregulation of cancerous inhibitor of protein phosphatase 2A (CIP2A), increased protein phosphatase 2A (PP2A) activity, and decreased AKT phosphorylation, but minimal effects on EGFR phosphorylation. Overexpression of CIP2A partially protected the H460 cells from TD-19-induced apoptosis. Okadaic acid, a known PP2A inhibitor, significantly reduced TD-19-induced apoptosis, while forskolin, which increased PP2A activity, increased the apoptotic effect of TD-19. TD-19 inhibited the growth of H460 xenograft tumors by ?80%. We conclude that TD-19 exerted tumoricidal effects on NSCLC cells. TD-19 provides proof that the CIP2A pathway may be a novel target for the treatment of EGFR wild-type NSCLC.
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Increased plasma soluble CD40 ligand concentration in pelvic inflammatory disease.
Clin. Chim. Acta
PUBLISHED: 09-02-2014
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The role of soluble CD40 ligand (sCD40L) in pelvic inflammatory disease (PID) remains unclear. We sought to determine whether sCD40L was an efficient serum marker as with WBC and CRP in PID patients.
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Macrophage migration inhibitory factor triggers chemotaxis of CD74+CXCR2+ NKT cells in chemically induced IFN-?-mediated skin inflammation.
J. Immunol.
PUBLISHED: 08-29-2014
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IFN-? mediates chemically induced skin inflammation; however, the mechanism by which IFN-?-producing cells are recruited to the sites of inflammation remains undefined. Secretion of macrophage migration inhibitory factor (MIF), a proinflammatory cytokine, from damaged cells may promote immune cell recruitment. We hypothesized that MIF triggers an initial step in the chemotaxis of IFN-?-producing cells in chemically induced skin inflammation. Using acute and chronic models of 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin inflammation in mouse ears, MIF expression was examined, and its role in this process was investigated pharmacologically. The cell populations targeted by MIF, their receptor expression patterns, and the effects of MIF on cell migration were examined. TPA directly caused cytotoxicity accompanied by MIF release in mouse ear epidermal keratinocytes, as well as in human keratinocytic HaCaT cells. Treatment with the MIF antagonist (S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid methyl ester considerably attenuated TPA-induced ear swelling, leukocyte infiltration, epidermal cell proliferation, and dermal angiogenesis. Inhibition of MIF greatly diminished the dermal infiltration of IFN-?(+) NKT cells, whereas the addition of exogenous TPA and MIF to NKT cells promoted their IFN-? production and migration, respectively. MIF specifically triggered the chemotaxis of NKT cells via CD74 and CXCR2, and the resulting depletion of NKT cells abolished TPA-induced skin inflammation. In TPA-induced skin inflammation, MIF is released from damaged keratinocytes and then triggers the chemotaxis of CD74(+)CXCR2(+) NKT cells for IFN-? production.
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Anti-adhesive effect of hyaluronate in a rabbit laminectomy model.
Biomed J
PUBLISHED: 08-14-2014
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Postlaminectomy dural adhesion is a common cause of recurrent symptoms. Hyaluronic acid-based gel has been reported to reduce the incidence of postoperative adhesion in the peritoneal cavity; however, its effect on preventing postoperative scar formation at laminectomy sites is not yet known. The purpose of this study was to evaluate the anti-adhesive effect of hyaluronic acid-based gelatin after laminectomy, using a rabbit model.
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From voxels to knowledge: a practical guide to the segmentation of complex electron microscopy 3D-data.
J Vis Exp
PUBLISHED: 08-13-2014
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Modern 3D electron microscopy approaches have recently allowed unprecedented insight into the 3D ultrastructural organization of cells and tissues, enabling the visualization of large macromolecular machines, such as adhesion complexes, as well as higher-order structures, such as the cytoskeleton and cellular organelles in their respective cell and tissue context. Given the inherent complexity of cellular volumes, it is essential to first extract the features of interest in order to allow visualization, quantification, and therefore comprehension of their 3D organization. Each data set is defined by distinct characteristics, e.g., signal-to-noise ratio, crispness (sharpness) of the data, heterogeneity of its features, crowdedness of features, presence or absence of characteristic shapes that allow for easy identification, and the percentage of the entire volume that a specific region of interest occupies. All these characteristics need to be considered when deciding on which approach to take for segmentation. The six different 3D ultrastructural data sets presented were obtained by three different imaging approaches: resin embedded stained electron tomography, focused ion beam- and serial block face- scanning electron microscopy (FIB-SEM, SBF-SEM) of mildly stained and heavily stained samples, respectively. For these data sets, four different segmentation approaches have been applied: (1) fully manual model building followed solely by visualization of the model, (2) manual tracing segmentation of the data followed by surface rendering, (3) semi-automated approaches followed by surface rendering, or (4) automated custom-designed segmentation algorithms followed by surface rendering and quantitative analysis. Depending on the combination of data set characteristics, it was found that typically one of these four categorical approaches outperforms the others, but depending on the exact sequence of criteria, more than one approach may be successful. Based on these data, we propose a triage scheme that categorizes both objective data set characteristics and subjective personal criteria for the analysis of the different data sets.
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Autophagy-preferential degradation of MIR224 participates in hepatocellular carcinoma tumorigenesis.
Autophagy
PUBLISHED: 07-25-2014
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Autophagy and microRNA (miRNA) are important regulators during cancer cell tumorigenesis. Impaired autophagy and high expression of the oncogenic microRNA MIR224 are prevalent in hepatocellular carcinoma (HCC); however, the relationship between the 2 phenomena remains elusive. In this study, we are the first to reveal that autophagy selectively regulates MIR224 expression through an autophagosome-mediated degradation system. Based on this finding, we further demonstrated that in hepatitis B virus (HBV)-related HCC, aberrant autophagy (low autophagic activity) results in accumulation of MIR224 and decreased expression of the target gene Smad4, which leads to increased cell migration and tumor formation. Preferential recruitment of MIR224 into the autophagosome was clearly demonstrated by a) miRNA in situ hybridization under confocal microscopy, and b) immunogold labeling of MIR224 under electron microscopy compared with a ubiquitously expressed microRNA MIRlet7e/let-7. Furthermore, we found that off-label use of amiodarone, an antiarrhythmic agent, effectively suppressed HCC tumorigenesis through autophagy-mediated MIR224 degradation both in vitro and in vivo. In summary, we identified amiodarone as a new autophagy inducer, which may provide an alternative approach in HCC therapy through a novel tumor suppression mechanism.
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Correlation of plasma osteopontin and neutrophil gelatinase-associated lipocalin levels with the severity and clinical outcome of pelvic inflammatory disease.
Taiwan J Obstet Gynecol
PUBLISHED: 07-15-2014
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To investigate the correlation of two important inflammatory biomarkers, plasma osteopontin and neutrophil gelatinase-associated lipocalin (NGAL), with the severity and outcome of pelvic inflammatory disease (PID).
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Isolation of a Bacteriophage and Its Depolymerase Specific for K1 Capsule of Klebsiella pneumoniae: Implication in Typing and Treatment.
J. Infect. Dis.
PUBLISHED: 07-07-2014
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Klebsiella pneumoniae causing community-acquired pyogenic liver abscess complicated with metastatic meningitis and endophthalmitis has emerged recently, most frequently associated with the K1 capsular type.
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Suppressing the relaxation oscillation noise of injection-locked WRC-FPLD for directly modulated OFDM transmission.
Opt Express
PUBLISHED: 07-01-2014
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By up-shifting the relaxation oscillation peak and suppressing its relative intensity noise in a weak-resonant-cavity Fabry-Perot laser diode (WRC-FPLD) under intense injection-locking, the directly modulated transmission of optical 16 quadrature amplitude modulation (QAM) orthogonal frequency division multiplexing (OFDM) data-stream is demonstrated. The total bit rate of up to 20 Gbit/s within 5-GHz bandwidth is achieved by using the OFDM subcarrier pre-leveling technique. With increasing the injection-locking power from -12 to -3 dBm, the effective reduction on threshold current of the WRC-FPLD significantly shifts its relaxation oscillation frequency from 5 to 7.5 GHz. This concurrently induces an up-shift of the peak relative intensity noise (RIN) of the WRC-FPLD, and effectively suppresses the background RIN level to -104 dBc/Hz within the OFDM band between 3 and 6 GHz. The enhanced signal-to-noise ratio from 16 to 20 dB leads to a significant reduction of bit-error-rate (BER) of the back-to-back transmitted 16-QAM-OFDM data from 1.3 × 10(-3) to 5 × 10(-5), which slightly degrades to 1.1 × 10(-4) after 25-km single-mode fiber (SMF) transmission. However, the enlarged injection-locking power from -12 to -3 dBm inevitably declines the modulation throughput and increases its negative throughput slope from -0.8 to -1.9 dBm/GHz. After pre-leveling the peak amplitude of the OFDM subcarriers to compensate the throughput degradation of the directly modulated WRC-FPLD, the BER under 25-km SMF transmission can be further improved to 3 × 10(-5) under a receiving power of -3 dBm.
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Gene therapy in patient-specific stem cell lines and a preclinical model of retinitis pigmentosa with membrane frizzled-related protein defects.
Mol. Ther.
PUBLISHED: 05-23-2014
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Defects in Membrane Frizzled-related Protein (MFRP) cause autosomal recessive retinitis pigmentosa (RP). MFRP codes for a retinal pigment epithelium (RPE)-specific membrane receptor of unknown function. In patient-specific induced pluripotent stem (iPS)-derived RPE cells, precise levels of MFRP, and its dicistronic partner CTRP5, are critical to the regulation of actin organization. Overexpression of CTRP5 in naïve human RPE cells phenocopied behavior of MFRP-deficient patient RPE (iPS-RPE) cells. AAV8 (Y733F) vector expressing human MFRP rescued the actin disorganization phenotype and restored apical microvilli in patient-specific iPS-RPE cell lines. As a result, AAV-treated MFRP mutant iPS-RPE recovered pigmentation and transepithelial resistance. The efficacy of AAV-mediated gene therapy was also evaluated in Mfrp(rd6)/Mfrp(rd6) mice--an established preclinical model of RP--and long-term improvement in visual function was observed in AAV-Mfrp-treated mice. This report is the first to indicate the successful use of human iPS-RPE cells as a recipient for gene therapy. The observed favorable response to gene therapy in both patient-specific cell lines, and the Mfrp(rd6)/Mfrp(rd6) preclinical model suggests that this form of degeneration caused by MFRP mutations is a potential target for interventional trials.
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Cisd2 modulates the differentiation and functioning of adipocytes by regulating intracellular Ca2+ homeostasis.
Hum. Mol. Genet.
PUBLISHED: 05-08-2014
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CISD2 is a causative gene associated with Wolfram syndrome (WFS). However, it remains a mystery as to how the loss of CISD2 causes metabolic defects in patients with WFS. Investigation on the role played by Cisd2 in specific cell types may help us to resolve these underlying mechanisms. White adipose tissue (WAT) is central to the maintenance of energy metabolism and glucose homeostasis in humans. In this study, adipocyte-specific Cisd2 knockout (KO) mice showed impairment in the development of epididymal WAT (eWAT) in the cell autonomous manner. A lack of Cisd2 caused defects in the biogenesis and function of mitochondria during differentiation of adipocytes in vitro. Insulin-stimulated glucose uptake and secretion of adiponectin by the Cisd2 KO adipocytes were decreased. Moreover, Cisd2 deficiency increased the cytosolic level of Ca(2+) and induced Ca(2+)-calcineurin-dependent signaling that inhibited adipogenesis. Importantly, Cisd2 was found to interact with Gimap5 on the mitochondrial and ER membranes and thereby modulate mitochondrial Ca(2+) uptake associated with the maintenance of intracellular Ca(2+) homeostasis in adipocytes. Thus, it would seem that Cisd2 plays an important role in intracellular Ca(2+) homeostasis, which is required for the differentiation and functioning of adipocytes as well as the regulation of glucose homeostasis in mice.
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Risk evaluation for the development of cervical intraepithelial neoplasia: Development and validation of risk-scoring schemes.
Int. J. Cancer
PUBLISHED: 04-25-2014
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Cervical cancer screening guidelines do not comprehensively define what constitutes high risk. This study developed and validated simple risk-scoring schemes to improve Papanicolaou smear screening for women at high risk. Four cumulative risk score (CRS) schemes were derived respectively for the development of cervical intraepithelial neoplasia grade 1 (CIN1) and grade 2 or worse (CIN2+) using community-based case-control data (n?=?1523). By calculating the area under the receiver operating characteristic (AU-ROC) curve, these schemes were validated in a Papanicolaou smear follow-up cohort (n?=?967) and a hospital-based cytology screening population (n?=?217). A high DNA load of high-risk human papillomavirus (HR-HPV) was the main predictor for CIN1 and CIN2+, although age, married status combined with the number of sexual partners, active and passive smoking and age at sexual debut also affected associated lesions. In the training set, only the HPV-testing-contained CIN2+ CRS scheme presented an excellent discrimination for identifying CIN2+ (AU-ROC?=?0.866). Using a CRS cutoff value of 4 to identify CIN2+, the sensitivity and specificity of predicting CIN2+ for the 3- and 5-year follow-ups were 100% and 90.8%, and 83.3% and 90.4%, respectively, in the validation cohort. In the hospital-based validation population, the CRS scheme showed comparable discrimination for CIN2+ detection (sensitivity 88.2% and specificity 84.6%). Women with CRS ?4 had a 5.4% and 9.1% of 3- and 5-year cumulative incidence, respectively, and a 40.5-fold hazard ratio of developing CIN2+. In conclusion, combined with HR-HPV testing and verified risk factors, a simple CRS scheme could effectively improve the implementation of CIN2+ screening.
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TIMP-3 -1296 T>C and TIMP-4 -55 T>C gene polymorphisms play a role in the susceptibility of hepatocellular carcinoma among women.
Tumour Biol.
PUBLISHED: 04-24-2014
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The purpose of this study was to investigate genetic impact of TIMP-3 -1296 T>C (rs9619311) and TIMP-4 -55 T>C (rs3755724) gene polymorphisms on the susceptibility and clinicopathological characteristics of hepatocellular carcinoma (HCC). A total of 759 subjects, including 530 healthy controls and 229 patients with hepatocellular carcinoma, were recruited in this study. Allelic discrimination of TIMP-3 -1296 T>C (rs9619311) and TIMP-4 -55 T>C (rs3755724) polymorphisms was assessed with the ABI StepOne™ Real-Time PCR System. Among women group, individuals with TC or CC alleles of TIMP-3 -1296 T>C gene polymorphism protected against HCC (AOR?=?0.35, 95% confidence interval (CI)?=?0.12-0.97; p?=?0.04) compared to individuals with TT alleles, after adjusting for other confounders. Also, women with TC alleles and with TC or CC alleles of TIMP-4 -55 T>C polymorphisms had a 2.52-fold risk (95%CI?=?1.23-5.13; p?=?0.01) and 2.47-fold risk (95%CI?=?1.26-4.87; p?=?0.008) of developing HCC compared to individuals with TT alleles, after adjusting for other confounders. There was no synergistic effect between gene polymorphism and environmental risk factors, including tobacco and alcohol consumptions and clinical statuses of HCC as well as serum expression of liver-related clinicopathological markers. In conclusion, gene polymorphisms of TIMP-3 -1296 T>C (rs9619311) and TIMP-4 -55 T>C (rs3755724) play a role in the susceptibility of HCC among Taiwan women.
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Application of multiscale entropy in arterial waveform contour analysis in healthy and diabetic subjects.
Med Biol Eng Comput
PUBLISHED: 04-19-2014
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We applied multiscale entropy (MSE) to assess variation in crest time (CT), a parameter in arterial waveform analysis, in diagnosing patients with diabetes. Data on digital volume pulse were obtained from 93 individuals in three groups [Healthy young (Group 1, 20< age ?40, n = 30), healthy upper-middle-aged (Group 2, age >40, n = 30), and diabetic (Group 3, n = 33) subjects]. Crest time, normalized crest time, crest time ratio (CTR), small- and large-scale MSE on CT [MSESS(CT) and MSELS(CT), respectively] were computed and correlated with anthropometric (i.e., body weight/height, waist circumference), hemodynamic (i.e., blood pressure), and biochemical parameters (i.e., serum triglyceride, high-density lipoprotein, fasting blood sugar, and glycosylated hemoglobin). The results demonstrated higher variability in CT in healthy subjects (Groups 1 and 2) compared with that in diabetic patients (Group 3) as reflected in significantly elevated MSESS(CT) and MSELS(CT) in the former (p < 0.003 and p < 0.001, respectively). MSELS(CT) also showed significant association with waist circumference and fasting blood sugar (i.e., two diagnostic criteria of metabolic syndrome) as well as glycosylated hemoglobin concentration. In conclusion, using MSE analysis for assessing CT variation successfully distinguished diabetic patients from healthy subjects. MSESS(CT) and MSELS(CT) therefore may serve as noninvasive tools for identifying subjects with diabetes and those at risk.
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Mechanotransduction in intervertebral discs.
J. Cell. Mol. Med.
PUBLISHED: 04-17-2014
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Mechanotransduction plays a critical role in intracellular functioning-it allows cells to translate external physical forces into internal biochemical activities, thereby affecting processes ranging from proliferation and apoptosis to gene expression and protein synthesis in a complex web of interactions and reactions. Accordingly, aberrant mechanotransduction can either lead to, or be a result of, a variety of diseases or degenerative states. In this review, we provide an overview of mechanotransduction in the context of intervertebral discs, with a focus on the latest methods of investigating mechanotransduction and the most recent findings regarding the means and effects of mechanotransduction in healthy and degenerative discs. We also provide some discussion of potential directions for future research and treatments.
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Influence of lumbar curvature and rotation on forward flexibility in idiopathic scoliosis.
Biomed J
PUBLISHED: 04-16-2014
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Lumbar spine facet joints are arranged sagittally and mainly provide forward flexibility. Rotation of the lumbar vertebral body and coronal plane deformity may influence the function of lumbar forward flexibility. We hypothesize that the more advanced axial and coronal plane deformity could cause more limitation on forward flexibility in patients with idiopathic scoliosis.
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Featured Article: Interactions of surface-expressed TLR-4 and endosomal TLR-9 accelerate lupus progression in anti-dsDNA antibody transgenic mice.
Exp. Biol. Med. (Maywood)
PUBLISHED: 04-09-2014
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The hallmark of systemic lupus erythematosus (SLE) is the presence of high levels of anti-double-stranded DNA autoantibody (anti-dsDNA) in sera. In addition, pathogen infections coincide frequently with the occurrence of lupus. Our study was designed to investigate the contribution of anti-dsDNA, extracellular and intracellular Toll-like receptors (TLRs), a family of pattern-recognition receptors for sensing invading pathogens, in the pathogenesis of lupus. Although cell surface-expressed TLR4 may promote lupus progression, intracellular nucleic acid-sensing TLR9 plays either stimulatory or protective roles in different murine lupus models. To examine the role of TLR4, TLR9, and anti-dsDNA in SLE, we generated transgenic mice carrying anti-dsDNA antibody transgene and challenged the mice with TLR4- and TLR9-agonists, lipopolysaccharides (LPS), and CpG oligodeoxynucleotide (CpG ODN1826 and 2216), respectively. Splenocytes from these mice were found to secrete higher levels of interleukin-10 (IL-10) and anti-dsDNA when treated with a combination of TLR4 and TLR9 agonists (LPS?+?CpG). In addition, the transgenic mice were intraperitoneally administered with CpG or combined CpG and LPS to determine whether extracellular TLR4 and intracellular TLR9 activations could affect lupus progression in vivo. It was found that serum levels of anti-dsDNA antibodies and interferon-alpha were higher in CpG?+?LPS-treated transgenic mice than those in non-transgenic mice. Besides, elevated levels of proteinuria, blood urine nitrogen, and immune complex depositions in kidney were found in treated transgenic mice. Anti-dsDNA and simultaneous activation of surface-expressed TLR4 and endosomal TLR9 are crucial to promote the lupus progression.
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Postoperative meningitis after spinal surgery: a review of 21 cases from 20,178 patients.
BMC Infect. Dis.
PUBLISHED: 04-07-2014
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Postoperative bacterial meningitis is a rare complication of spinal surgery and is considered to be a complication related to intraoperative incidental durotomy. A high index of suspicion for meningitis is essential in patients who have the clinical triad of fever, neck stiffness and consciousness disturbance during the postoperative period. A delay in diagnosis or treatment can lead to morbidity and mortality. Due to the low incidence of postoperative meningitis, very few studies have reported this complication. The purpose of this study was to report the clinical features, laboratory evaluations, treatment course and prognosis of 21 patients with post spinal surgery meningitis.
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Symptomatic epidural hematoma after lumbar decompression surgery.
Eur Spine J
PUBLISHED: 03-28-2014
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Postoperative symptomatic epidural hematoma (SEH) is a serious complication of lumbar spine surgery. Despite its rarity, this uncommon complication may result in devastating neurological sequelae, including lower limb weakness.
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Development of optical probes for in vivo imaging of polarized macrophages during foreign body reactions.
Acta Biomater
PUBLISHED: 03-27-2014
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Plasticity of macrophage (M?) phenotypes exist in a spectrum from classically activated (M1) cells, to alternatively activated (M2) cells, contributing to both the normal healing of tissues and the pathogenesis of implant failure. Here, folate- and mannose-based optical probes were fabricated to simultaneously determine the degree of M? polarization. In vitro tests show the ability of these probes to specifically target M1 and M2 cells. In an in vivo murine model, they were able to distinguish between the M1-dominated inflammatory response to infection and the M2-dominated regenerative response to particle implants. Finally, the probes were used to assess the inflammatory/regenerative properties of biomaterial implants. Our results show that these probes can be used to monitor and quantify the dynamic processes of M? polarization and their role in cellular responses in real time.
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Surgical risks and perioperative complications of instrumented lumbar surgery in patients with liver cirrhosis.
Biomed J
PUBLISHED: 03-27-2014
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Patients with liver cirrhosis have high surgical risks due to malnutrition, impaired immunity, coagulopathy, and encephalopathy. However, there is no information in English literature about the results of liver cirrhotic patients who underwent instrumented lumbar surgery. The purpose of this study is to report the perioperative complications, clinical outcomes and determine the surgical risk factors in cirrhotic patients.
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Emergency endovascular aortic repair of a ruptured mycotic aorto-iliac aneurysm presenting with lumbar radiculopathy.
Cardiovasc J Afr
PUBLISHED: 03-09-2014
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Ruptured abdominal aortic aneurysm is life-threatening without immediate management. The initial clinical presentation is non-specific and impending rupture is easily missed, especially without a CT scan. We present a case of a 56-year-old man with low-back pain and left lower-extremity numbness, which was diagnosed as a herniated intervertebral disc (HIVD) with left acute sciatica syndrome. He also complained of persistent fever and abdominal discomfort. Routine blood work-up revealed leukocytosis and decreasing haemoglobin levels. CT angiography (CTA) showed impending rupture of the left aorto-iliac aneurysm. We therefore performed endovascular aneurysm repair (EVAR). Blood culture revealed Salmonella enterica, for which he received antibiotics. No acute sciatica syndrome was present immediately after the EVAR. No EVAR-related complications were noted in the one-year CTA follow up.
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Twist1-induced dissemination preserves epithelial identity and requires E-cadherin.
J. Cell Biol.
PUBLISHED: 03-05-2014
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Dissemination of epithelial cells is a critical step in metastatic spread. Molecular models of dissemination focus on loss of E-cadherin or repression of cell adhesion through an epithelial to mesenchymal transition (EMT). We sought to define the minimum molecular events necessary to induce dissemination of cells out of primary murine mammary epithelium. Deletion of E-cadherin disrupted epithelial architecture and morphogenesis but only rarely resulted in dissemination. In contrast, expression of the EMT transcription factor Twist1 induced rapid dissemination of cytokeratin-positive epithelial cells. Twist1 induced dramatic transcriptional changes in extracellular compartment and cell-matrix adhesion genes but not in cell-cell adhesion genes. Surprisingly, we observed disseminating cells with membrane-localized E-cadherin and ?-catenin, and E-cadherin knockdown strongly inhibited Twist1-induced single cell dissemination. Dissemination can therefore occur with retention of epithelial cell identity. The spread of cancer cells during metastasis could similarly involve activation of an epithelial motility program without requiring a transition from epithelial to mesenchymal character.
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Effects of urotensin II on intracellular pH regulation in cultured human internal mammary artery smooth muscle cells.
Peptides
PUBLISHED: 03-01-2014
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The Na(+)-H(+) exchanger (NHE) and the Na(+)-HCO3(-) co-transporter (NBC) have been confirmed as two major active acid extruders in many mammalian cells. Whether the NHE and NBC functional co-exist in human internal mammary artery smooth muscle cells (HIMASMCs) remains unclear. The aims of the present study were to investigate the acid-extruding mechanisms and to explore the effects of urotensin-II (U-II), a powerful vasoconstrictor, on pHi regulators in HIMASMCs. We investigated the changes of pHi by BCECF-fluorescence in HIMASMCs. We found that (a) two Na(+)-dependent acid extruders, i.e. NHE and NBC, functionally co-exist; (b) U-II (3-100 nM) induced a concentration-dependent intracellular acidosis; and (c) U-II (3-100 nM) caused a concentration-dependent increase on NHE activity, while decrease on NBC activity. In summary, we demonstrate for the first time that two acid-extruders, NHE and NBC, functionally co-exist in HIMASMCs. Moreover, U-II induces a concentration-dependent intracellular acidosis through the balanced effect of its effect on increasing NHE activity and decreasing NBC activity.
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Association of novel gene polymorphisms RRM1 -756T>C and -269 C>A with breast cancer.
J. Clin. Lab. Anal.
PUBLISHED: 02-27-2014
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Genetic variants are reported to play an important role in the susceptibility of breast cancer. Ribonucleotide reductase 1 (RRM1) is suggested to play an essential role in the regulation of cancer development. The purpose of this study was to identify novel gene polymorphisms of RRM1 -756T>C and RRM1 -269 C>A specific to patients with breast cancer and healthy controls.
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CIP2A mediates erlotinib-induced apoptosis in non-small cell lung cancer cells without EGFR mutation.
Lung Cancer
PUBLISHED: 02-25-2014
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Epidermal growth factor receptor (EGFR) inhibitors show favorable clinical response in some patients with non-small cell lung cancer (NSCLC) who have no EGFR mutation, indicating alternative mechanisms for their tumoricidal effects. We previously showed erlotinib, a selective EGFR antagonist, inhibited the growth of sensitive hepatocellular carcinoma cells by inhibiting the cancerous inhibitor of protein phosphatase 2A (CIP2A) pathway. The aim of this study was to determine if erlotinib can also inhibit the growth of NSCLC cells by inactivating the CIP2A-dependent signaling pathway.
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Expression of growth arrest-specific protein 6 and Axl molecules in the left internal mammary artery of patients undergoing coronary artery bypass grafting.
J. Clin. Pathol.
PUBLISHED: 02-24-2014
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Studies have demonstrated that using a left internal mammary artery (LIMA) graft yields excellent long-term results in coronary artery bypass grafting (CABG). The growth arrest-specific 6 (Gas6) gene and its receptor, Axl, are crucial in vascular haemostasis and atherosclerosis. The objective of this study was to determine the expression of Gas6 and Axl molecules in the aorta and LIMA in patients undergoing CABG.
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Alternative strategies to manipulate fibrocyte involvement in the fibrotic tissue response: pharmacokinetic inhibition and the feasibility of directed-adipogenic differentiation.
Acta Biomater
PUBLISHED: 02-17-2014
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Fibrocytes have previously been identified as important mediators in several inflammatory and fibrotic diseases. However, there is no effective treatment thus far to reduce fibrotic tissue responses without affecting wound healing reactions. Here we investigate two strategies to alleviate fibrocyte interactions at the biomaterial interface, reducing collagen production and scar tissue formation. First, in an indirect approach, TGF-? inhibitor-SB431542 and IL-1?/TNF-? inhibitor SB203580 were locally released from scaffold implants to block their respective signaling pathways. We show that the inhibition of IL-1?/TNF-? has no influence on overall fibrotic tissue reactions to the implants. However, the reduction of localized TGF-? significantly decreases the fibrocyte accumulation and myofibroblast activation while reducing the fibrotic tissue formation. Since fibrocytes can be differentiated into non-fibrotic cell types, such as adipocytes, we further sought a more direct approach to reduce fibrocyte responses by directing fibrocyte differentiation into adipocytes. Interestingly, by initiating fibrocyte-to-adipocyte differentiation through sustained differentiation cocktail release, we find that adipogenic differentiation forces incoming fibrocytes away from the traditional myofibroblast lineage, leading to a substantial reduction in the collagen formation and fibrotic response. Our results support a novel and effective strategy to improve implant safety by reducing implant-associated fibrotic tissue reactions via directing non-fibrotic differentiation of fibrocytes.
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Function of DNA methyltransferase 3a in lead (Pb(2+) )-Induced Cyclooxygenase-2 gene.
Environ. Toxicol.
PUBLISHED: 02-12-2014
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Lead ions (Pb(2+) ) are toxic industrial pollutants associated with chronic inflammatory diseases in humans and animals. Previously, we found that Pb(2+) ions induce COX-2 gene expression via the EGF receptor/nuclear factor-?B signal transduction pathway in epidermoid carcinoma cell line A431. In this study, to see whether Pb(2+) ions affect COX-2 expression by epigenetic mechanisms, we looked at the mRNAs of DNA methyltransferases (DNMTs) using real-time PCR of total RNA from these cells. Cells exposed to Pb(2+) had low levels of DNMT3a mRNA, whereas the levels of DNMT1 and DNMT3b mRNAs remained unchanged. Pretreatment of cells with DNMT inhibitor 5-aza-2'-deoxycytidine (5 ?M) followed by Pb(2+) (1 ?M) significantly increased levels of COX-2 mRNA compared with cells treated with Pb(2+) alone. Overexpression of tumor suppressor gene Rb correlated with an increase in COX-2 mRNA and a decrease in DNMT3a mRNA. Conversely, overexpression of transcription factor E2F1 correlated with a decrease in COX-2 mRNA and an increase in DMNT3a mRNA. Pretreatment with EGFR inhibitors AG1478 and PD153035 significantly limited Pb(2+) -induced reduction in DNMT3a mRNA. In addition, gene knockdown of DNMT3a with short hairpin RNA correlated with increased COX-2 mRNA induced by Pb(2+) . Our findings suggest Pb(2+) ions induce COX-2 expression indirectly by reducing DNMT3a methylation of the COX-2 promoter via transcription factors Rb and E2F1. © 2014 Wiley Periodicals, Inc. Environ Toxicol, 2014.
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Validation of genome-wide association study (GWAS)-identified disease risk alleles with patient-specific stem cell lines.
Hum. Mol. Genet.
PUBLISHED: 02-04-2014
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While the past decade has seen great progress in mapping loci for common diseases, studying how these risk alleles lead to pathology remains a challenge. Age-related macular degeneration (AMD) affects 9 million older Americans, and is characterized by the loss of the retinal pigment epithelium (RPE). Although the closely linked genome-wide association studies ARMS2/HTRA1 genes, located at the chromosome 10q26 locus, are strongly associated with the risk of AMD, their downstream targets are unknown. Low population frequencies of risk alleles in tissue banks make it impractical to study their function in cells derived from autopsied tissue. Moreover, autopsy eyes from end-stage AMD patients, where age-related RPE atrophy and fibrosis are already present, cannot be used to determine how abnormal ARMS2/HTRA1 expression can initiate RPE pathology. Instead, induced pluripotent stem (iPS) cell-derived RPE from patients provides us with earlier stage AMD patient-specific cells and allows us to analyze the underlying mechanisms at this critical time point. An unbiased proteome screen of A2E-aged patient-specific iPS-derived RPE cell lines identified superoxide dismutase 2 (SOD2)-mediated antioxidative defense in the genetic allele's susceptibility of AMD. The AMD-associated risk haplotype (T-in/del-A) impairs the ability of the RPE to defend against aging-related oxidative stress. SOD2 defense is impaired in RPE homozygous for the risk haplotype (T-in/del-A; T-in/del-A), while the effect was less pronounced in RPE homozygous for the protective haplotype (G-Wt-G; G-Wt-G). ARMS2/HTRA1 risk alleles decrease SOD2 defense, making RPE more susceptible to oxidative damage and thereby contributing to AMD pathogenesis.
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Phenothiazine-modified electrodes: a useful platform for protein adsorption study.
Langmuir
PUBLISHED: 02-04-2014
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Using glucose oxidase (GOx) as a target protein, we studied the adsorption of protein on the phenothiazine-modified electrodes and assessed the potential of using the electrodes in biochemical applications. Experiment results showed that thionine chloride (TC) and its structural analogues, such as toluidine blue and methylene blue, fluoresced under photochemical excitation after being immobilized on indium-doped tin oxide (ITO) electrodes fabricated using either diazotization-reduction or oxidative polymerization. The surface-bound phenothiazines exhibited substantial binding affinities to the protein. At a pH > 5, the adsorbate showed no sign of desorption even the electrodes were electrically biased with voltages between ±0.3 V vs SCE. Thus, emission decay occurred while GOx was injected over the electrodes, which was consistent with the observations made using conductive-mode atomic force microscopy (CM-AFM). Under a quiescent condition, the protein interacted with the immobilized TC via a pseudo-first-order kinetic mechanism. The reaction reached a maximum rate at a pH > 5, at which the rate constant was approximately 7 × 10(-8) L/(U s). Under this condition, the adsorption rate increased as the level of the protein increased, regardless of pH, revealing application potential for GOx quantitation. The adsorption rate, however, decreased with a decrease in pH if the pH < 5. We concluded that static interactions played a crucial role. By monitoring Fe(CN)6(3-/4-) taking place at the TC-modified electrodes in pH 7 solutions, we observed that the adsorption of GOx imposed impedance on Fe(CN)6(3-/4-). The resulting charge-transfer resistance (RCT) increased as the amount of the protein increased, leading to a conclusion that the protein could reach the maximum surface coverage when its concentrations were greater than 100 U/mL. The protein molecules were likely repel each other as approaching the TC sites. Despite this, they maintained the native bioactivity after being adsorbed, enabling the TC-modified electrodes to function as glucose sensors. Glucose concentrations between 1 and 60 mM could be detected. Long-term analysis, in addition, showed that the electrode responses to the analyte were consistent and reproducible. Phenothiazine-modified electrodes are evident as a useful tool for understanding the adsorption of protein on solid surfaces and quantifying proteins.
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Decreased expression of organic cation transporters, Oct1 and Oct2, in brain microvessels and its implication to MPTP-induced dopaminergic toxicity in aged mice.
J. Cereb. Blood Flow Metab.
PUBLISHED: 02-03-2014
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This study was to investigate the influence of age on the expression of organic cation transporters (OCTs) that belong to the SLC22 family in brain microvessels (BMVs) and its implications for 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced dopaminergic toxicity in mice. Here, we showed that Oct1 and Oct2, but not Oct3, mRNAs were detected and enriched (compared with cerebral cortex) in BMVs of C57BL/6 (B6) mice using reverse transcription-quantitative PCR (RT-qPCR), and immunofluorescence analysis further revealed that Oct1 and Oct2 proteins were colocalized with endothelial markers. Both the mRNA and protein levels of Oct1 and Oct2 were reduced in aged mice. After an intraperitoneal administration of MPTP, brain extracellular levels of MPTP and 1-methyl-4-phenyl-pyridinium (MPP(+)) were much lower in aged mice and in Oct1/2(-/-) mice compared with younger mice and wild-type control mice, respectively. Knockout of Oct1/Oct2 protected Oct1/2(-/-) mice from MPTP-induced neurotoxicity, whereas the loss of tyrosine hydroxylase (TH)-positive neurons was slightly greater in aged than in younger mice. However, intrastriatal infusion of low-dose MPTP caused more severe dopaminergic toxicity in the substantia nigra of both aged mice and Oct1/2(-/-) mice. These findings show that age-dependent downregulation or knockout of Oct1/Oct2 in BMVs may reduce the transport of MPTP, which, in part, affects its dopaminergic toxicity.Journal of Cerebral Blood Flow & Metabolism advance online publication, 24 September 2014; doi:10.1038/jcbfm.2014.162.
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New markers in pelvic inflammatory disease.
Clin. Chim. Acta
PUBLISHED: 01-29-2014
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Pelvic inflammatory disease (PID) is a common infection in women of reproductive age. However, diagnosis of PID can be difficult due to the wide variation in the symptoms and signs, ranging from subtle or mild symptoms to severe pain in the lower abdomen. Clinical diagnosis alone has only 87% sensitivity and 50% specificity. Therefore, identifying biological factors that are useful for early diagnosis and correlating their expression with the severity of PID could provide significant benefits to women suffering from PID. Pentraxin 3 (PTX3), E-cadherin, myeloperoxidase, stromal cell-derived factor 1 (SDF-1) and the matrix metalloproteinase-9 (MMP-9)/MMP-2 ratio are potential candidates for detecting PID reliably. As PID is often subtle, highly sensitive PID detection methods are needed to promote the prevention of severe sequelae. Growth arrest-specific 6 (Gas6), in combination with its soluble tyrosine kinase receptor, sAxl, could elevate the sensitivity to 92%, which was higher than all other markers tested. Moreover, PTX3, D-dimer and YKL-40 concentrations can predict the clinical course of PID. Although single nucleotide polymorphisms of biomarker genes are not associated with the development of PID, myeloperoxidase SNP -463 G/A and SDF-1 SNP 801 G/A may affect the aggravated expression of their biomarkers in PID.
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The modified rice ?Amy8 promoter confers high-level foreign gene expression in a novel hypoxia-inducible expression system in transgenic rice seedlings.
Plant Mol. Biol.
PUBLISHED: 01-15-2014
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Expression of ?-amylase genes in rice is induced not only by sugar starvation and gibberellin (GA) but also by O2 deficiency. Promoters of two rice ?-amylase genes, ?Amy3 and ?Amy8, have been shown to direct high-level production of recombinant proteins in rice suspension cells and germinated seeds. In the present study, we modified the cis-acting DNA elements within the sugar/GA response complex (SRC/GARC) of ?Amy8 promoter. We found that addition of a G box and duplicated TA box leads to high-level expression of ?Amy8 SRC/GARC and significantly enhances ?Amy8 promoter activity in transformed rice cells and germinated transgenic rice seeds. We also show that these modifications have drastically increased the activity of ?Amy8 promoter in rice seedlings under hypoxia. Our results reveal that the G box and duplicated TA box may play important roles in stimulating promoter activity in response to hypoxia in rice. The modified ?Amy8 promoter was used to produce the recombinant human epidermal growth factor (hEGF) in rice cells and hypoxic seedlings. We found that the bioactive recombinant hEGF are stably produced and yields are up to 1.8% of total soluble protein (TSP) in transformed rice cells. The expression level of synthetic hEGF containing preferred rice codon usage comprises up to 7.8% of TSP in hypoxic transgenic seedlings. Our studies reveal that the modified ?Amy8 promoter can be applicable in establishing a novel expression system for the high-level production of foreign proteins in transgenic rice cells and seedlings under hypoxia.
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Antiarrhythmic effects of dehydroevodiamine in isolated human myocardium and cardiomyocytes.
J Ethnopharmacol
PUBLISHED: 01-14-2014
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Dehydroevodiamine alkaloid (DeHE), a bioactive component of the Chinese herbal medicine Wu-Chu-Yu (Evodiae frutus), exerted antiarrhythmic effect in guinea-pig ventricular myocytes. We further characterize the electromechanical effects of DeHE in the human atrial and ventricular tissues obtained from hearts of patients undergoing corrective cardiac surgery or heart transplantation.
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Generation of induced pluripotent stem cells from conjunctiva.
Graefes Arch. Clin. Exp. Ophthalmol.
PUBLISHED: 01-14-2014
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The objective of this study was to determine whether cells from the conjunctiva could be reprogrammed into induced pluripotent stem (iPS) cells, providing an alternative source of stem cells.
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Acute pulmonary injury with refractory hypoxaemia after implantation of Levitronix CentriMag ventricular assist device: successful treatment with veno-venous extracorporeal membrane oxygenation.
J Artif Organs
PUBLISHED: 01-03-2014
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Although acute pulmonary injury after cardiopulmonary bypass has been detailed in the literature, it was seldom mentioned in the context of following implantation of a ventricular assist device. We report on a 65-year-old male with end-stage ischemic cardiomyopathy who underwent implantation of Levitronix CentriMag (Levitronix, Waltham, MA) for cardiac support and was listed for heart transplantation. Acute pulmonary injury with profound hypoxaemia was noted 6 h after the implantation. Despite optimal medical treatment and maximal ventilator support, refractory hypoxaemia persisted, and veno-venous extracorporeal membrane oxygenation (oxygenator: Affinity-NT; centrifugal pump: BPX-80 Bio-Pump, Medtronic, Minneapolis, MN, USA) was applied for ventilation support. The patient was weaned from the extracorporeal membrane oxygenation 4 days later and from the ventilator on the next 2 days. He underwent a successful orthotopic heart transplant after a total of 77 days on Levitronix left ventricular device cardiac support.
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The Impact of SYNTAX Score of Non-Infarct-Related Artery on Long- Term Outcome among Patients with Acute ST Segment Elevation Myocardial Infarction Undergoing Primary Percutaneous Coronary Intervention.
PLoS ONE
PUBLISHED: 01-01-2014
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We investigated the impact of the severity of stenosis in a non-infarct-related artery (IRA) on the long-term prognosis of patients with ST-segment elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI).
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Urotensin II inhibits doxorubicin-induced human umbilical vein endothelial cell death by modulating ATF expression and via the ERK and Akt pathway.
PLoS ONE
PUBLISHED: 01-01-2014
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Regulation of the homeostasis of vascular endothelium is critical for the processes of vascular remodeling and angiogenesis under physiological and pathological conditions. Urotensin II (U-II), a potent vasoactive peptide, participates in vascular and myocardial remodeling after injury. We investigated the protective effect of U-II on doxorubicin (DOX)-induced apoptosis in cultured human umbilical vein endothelial cells (HUVECs) and the potential mechanisms involved in this process.
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Electron tomography of cryo-immobilized plant tissue: a novel approach to studying 3D macromolecular architecture of mature plant cell walls in situ.
PLoS ONE
PUBLISHED: 01-01-2014
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Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D) organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. In this paper we demonstrate that by cryo-immobilizing fresh tissue, then either cryo-sectioning or freeze-substituting and resin embedding, followed by cryo- or room temperature (RT) electron tomography, respectively, we can visualize previously unseen details of plant cell wall architecture in 3D, at macromolecular resolution (? 2 nm), and in near-native state. Qualitative and quantitative analyses showed that wall organization of cryo-immobilized samples were preserved remarkably better than conventionally prepared samples that suffer substantial extraction. Lignin-less primary cell walls were well preserved in both self-pressurized rapidly frozen (SPRF), cryo-sectioned samples as well as high-pressure frozen, freeze-substituted and resin embedded (HPF-FS-resin) samples. Lignin-rich secondary cell walls appeared featureless in HPF-FS-resin sections presumably due to poor stain penetration, but their macromolecular features could be visualized in unprecedented details in our cryo-sections. While cryo-tomography of vitreous tissue sections is currently proving to be instrumental in developing 3D models of lignin-rich secondary cell walls, here we confirm that the technically easier method of RT-tomography of HPF-FS-resin sections could be used immediately for routine study of low-lignin cell walls. As a proof of principle, we characterized the primary cell walls of a mutant (cob-6) and wild type Arabidopsis hypocotyl parenchyma cells by RT-tomography of HPF-FS-resin sections, and detected a small but significant difference in spatial organization of cellulose microfibrils in the mutant walls.
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Post-meal ?-cell function predicts the efficacy of glycemic control in patients with type 2 diabetes inadequately controlled by metformin monotherapy after addition of glibenclamide or acarbose.
Diabetol Metab Syndr
PUBLISHED: 01-01-2014
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This study aimed to explore parameters which will predict good control of HbA1c after adding a second anti-diabetic drug in patients with type 2 diabetes mellitus (T2DM) inadequately controlled with metformin monotherapy.
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The expression of ribonucleotide reductase M2 in the carcinogenesis of uterine cervix and its relationship with clinicopathological characteristics and prognosis of cancer patients.
PLoS ONE
PUBLISHED: 01-01-2014
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To investigate the implication of ribonucleotide reductase M2 (RRM2) in the carcinogenesis of uterine cervix and its relationship with clinicopathological characteristics and prognosis of cancer patients.
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Intracellular Acid-extruding regulators and the effect of lipopolysaccharide in cultured human renal artery smooth muscle cells.
PLoS ONE
PUBLISHED: 01-01-2014
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Homeostasis of the intracellular pH (pHi) in mammalian cells plays a pivotal role in maintaining cell function. Thus far, the housekeeping Na(+)-H(+) exchanger (NHE) and the Na(+)-HCO3(-) co-transporter (NBC) have been confirmed in many mammalian cells as major acid extruders. However, the role of acid-extruding regulators in human renal artery smooth muscle cells (HRASMCs) remains unclear. It has been demonstrated that lipopolysaccharide (LPS)-induced vascular occlusion is associated with the apoptosis, activating calpain and increased [Ca(2+)]i that are related to NHE1 activity in endothelia cells. This study determines the acid-extruding mechanisms and the effect of LPS on the resting pHi and active acid extruders in cultured HRASMCs. The mechanism of pHi recovery from intracellular acidosis (induced by NH4Cl-prepulse) is determined using BCECF-fluorescence in cultured HRASMCs. It is seen that (a) the resting pHi is 7.19 ± 0.03 and 7.10 ± 0.02 for HEPES- and CO2/HCO3(-)- buffered solution, respectively; (b) apart from the housekeeping NHE1, another Na(+)-coupled HCO3(-) transporter i.e. NBC, functionally co-exists to achieve acid-equivalent extrusion; (c) three different isoforms of NBC: NBCn1 (SLC4A7; electroneutral), NBCe1 (SLC4A4; electrogenic) and NBCe2 (SLC4A5), are detected in protein/mRNA level; and (d) pHi and NHE protein expression/activity are significantly increased by LPS, in both a dose- and time- dependent manner, but NBCs protein expression is not. In conclusion, it is demonstrated, for the first time, that four pHi acid-extruding regulators: NHE1, NBCn1, NBCe1 and NBCe2, co-exist in cultured HRASMCs. LPS also increases cellular growth, pHi and NHE in a dose- and time-dependent manner.
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The role of calpain-myosin 9-Rab7b pathway in mediating the expression of Toll-like receptor 4 in platelets: a novel mechanism involved in ?-granules trafficking.
PLoS ONE
PUBLISHED: 01-01-2014
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Toll-like receptors (TLRs) plays a critical role in innate immunity. In 2004, Aslam R. and Shiraki R. first determined that murine and human platelets express functional TLRs. Additionally, Andonegui G. demonstrated that platelets express TLR4, which contributes to thrombocytopenia. However, the underlying mechanisms of TLR4 expression by platelets have been rarely explored until now. The aim of this study was to identify the mechanism of TLR4 expression underlying thrombin treatment. The human washed platelets were used in this study. According to flowcytometry and western blot analysis, the surface levels of TLR4 were significantly enhanced in thrombin-activated human platelets and decreased by TMB-8, calpeptin, and U73122, but not Y27632 (a Rho-associated protein kinase ROCK inhibitor) indicating that thrombin-mediated TLR4 expression was modulated by PAR/PLC pathway, calcium and calpain. Co-immunoprecipitation (co-IP) assay demonstrated that the interaction between TLR4 and myosin-9 (a substrate of calpain) was regulated by calpain; cleavage of myosin-9 enhanced TLR4 expression in thrombin treated platelets. Transmission electron microscope data indicated that human platelets used ?-granules to control TLR4 expression; the co-IP experiment suggested that myosin-9 did not coordinate with Rab7b to negatively regulate TLR4 trafficking in thrombin treated platelets. In summary, phospholipase C?-calpain-myosin 9-Rab7b axis was responsible for the mechanism underlying the regulation of TLR4 containing ?-granules trafficking in thrombin-stimulated platelets, which was involved in coagulation.
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Interorganelle interactions and inheritance patterns of nuclei and vacuoles in budding yeast meiosis.
Autophagy
PUBLISHED: 12-04-2013
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Many of the mechanisms by which organelles are inherited by spores during meiosis are not well understood. Dramatic chromosome motion and bouquet formation are evolutionarily conserved characteristics of meiotic chromosomes. The budding yeast bouquet genes (NDJ1, MPS3, CSM4) mediate these movements via telomere attachment to the nuclear envelope (NE). Here, we report that during meiosis the NE is in direct contact with vacuoles via nucleus-vacuole junctions (NVJs). We show that in meiosis NVJs are assembled through the interaction of the outer NE-protein Nvj1 and the vacuolar membrane protein Vac8. Notably, NVJs function as diffusion barriers that exclude the nuclear pore complexes, the bouquet protein Mps3 and NE-tethered telomeres from the outer nuclear membrane and nuclear ER, resulting in distorted NEs during early meiosis. An increase in NVJ area resulting from Nvj1-GFP overexpression produced a moderate bouquet mutant-like phenotype in wild-type cells. NVJs, as the vacuolar contact sites of the nucleus, were found to undergo scission alongside the NE during meiotic nuclear division. The zygotic NE and NVJs were partly segregated into 4 spores. Lastly, new NVJs were also revealed to be synthesized de novo to rejoin the zygotic NE with the newly synthesized vacuoles in the mature spores. In conclusion, our results revealed that budding yeast nuclei and vacuoles exhibit dynamic interorganelle interactions and different inheritance patterns in meiosis, and also suggested that nvj1? mutant cells may be useful to resolve the technical challenges pertaining to the isolation of intact nuclei for the biochemical study of meiotic nuclear proteins.
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Localised bullous eruptions after extravasation of normal saline in the forearm during left ventricular device-assisted surgery.
Cardiovasc J Afr
PUBLISHED: 11-20-2013
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Peripheral infusion of intravenous agents is a daily routine in hospitals. Extravasation is an unintended complication associated with intravenous infusion where accidental injection or leakage of fluid occurs into the perivascular or subcutaneous space. Extravasation is fairly common but is usually without serious consequences. This has led clinicians to underestimate the potentially serious consequences of extravasation. Extravasation injury results from a combination of factors, including cytotoxicity of the solution, osmolality, vasoconstrictor effects, infusion pressure and other factors. We describe a case of upper extremity localised bullous eruptions resulting from the pressurised infusion of crystalloid solutions through an intravenous catheter, placed in the operating room during left ventricular device-assisted surgery. Peri-operative management of acute localised bullous eruptions requires surveillance for unforeseen consequences. Early recognition, diagnosis and intervention averted potential complications and morbidity.
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Using single-molecule approaches to study archaeal DNA-binding protein Alba1.
Biochemistry
PUBLISHED: 10-23-2013
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Thermophilic and hyperthermophilic archaea have one or more copies of the Alba gene, which encodes Alba, a dimeric, highly basic protein that binds cooperatively to DNA. However, the functions of Alba and how it interacts with DNA remain unclear. In this study, we have used single-molecule tethered particle motion (TPM) and optical tweezers (OT) experiments to study the interactions between DNA molecules and Alba1. When Alba1 binds to double-stranded DNA, the Brownian motion (BM) amplitude for DNA tethers increases continuously, suggesting that Alba1 binds cooperatively. The OT study confirmed that a 5-fold increase in the persistence length of the Alba1 nucleoprotein filament is the major factor causing the increase in the BM amplitude for DNA tethers, while the contour length remained mostly unchanged. Moreover, the rate of the increase in the BM amplitude and the BM plateau value are both DNA length-dependent, indicating that the number of Alba1 initiation binding sites increases as the DNA becomes longer. Using the incoming-strand TPM experiment to monitor the interaction between Alba1 nucleoprotein filaments, we found that significant dimer-dimer contacts between two Alba1 nucleoprotein filaments are present, and the interaction is regulated by the concentration of Alba1.
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Enhancement of ADP release from the RAD51 presynaptic filament by the SWI5-SFR1 complex.
Nucleic Acids Res.
PUBLISHED: 09-27-2013
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Homologous recombination catalyzed by the RAD51 recombinase eliminates deleterious DNA lesions from the genome. In the presence of ATP, RAD51 forms a nucleoprotein filament on single-stranded DNA, termed the presynaptic filament, to initiate homologous recombination-mediated DNA double-strand break repair. The SWI5-SFR1 complex stabilizes the presynaptic filament and enhances its ability to mediate the homologous DNA pairing reaction. Here we characterize the RAD51 presynaptic filament stabilization function of the SWI5-SFR1 complex using optical tweezers. Biochemical experiments reveal that SWI5-SFR1 enhances ATP hydrolysis by single-stranded DNA-bound RAD51. Importantly, we show that SWI5-SFR1 acts by facilitating the release of ADP from the presynaptic filament. Our results thus provide mechanistic understanding of the function of SWI5-SFR1 in RAD51-mediated DNA recombination.
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Optical imaging of fibrin deposition to elucidate participation of mast cells in foreign body responses.
Biomaterials
PUBLISHED: 09-03-2013
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Mast cell activation has been shown to be an initiator and a key determinant of foreign body reactions. However, there is no non-invasive method that can quantify the degree of implant-associated mast cell activation. Taking advantage of the fact that fibrin deposition is a hallmark of mast cell activation around biomaterial implants, a near infrared probe was fabricated to have high affinity to fibrin. Subsequent in vitro testing confirmed that this probe has high affinity to fibrin. Using a subcutaneous particle implantation model, we found significant accumulation of fibrin-affinity probes at the implant sites as early as 15 min following particle implantation. The accumulation of fibrin-affinity probes at the implantation sites could also be substantially reduced if anti-coagulant - heparin was administered at the implant sites. Further studies have shown that subcutaneous administration of mast cell activator - compound 48/80 - prompted the accumulation of fibrin-affinity probes. However, implant-associated fibrin-affinity probe accumulation was substantially reduced in mice with mast cell deficiency. The results show that our fibrin-affinity probes may serve as a powerful tool to monitor and measure the extent of biomaterial-mediated fibrin deposition and mast cell activation in vivo.
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Therapeutic margins in a novel preclinical model of retinitis pigmentosa.
J. Neurosci.
PUBLISHED: 08-16-2013
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The third-most common cause of autosomal recessive retinitis pigmentosa (RP) is due to defective cGMP phosphodiesterase-6 (PDE6). Previous work using viral gene therapy on PDE6-mutant mouse models demonstrated photoreceptors can be rescued if administered before degeneration. However, whether visual function can be rescued after degeneration onset has not been addressed. This is a clinically important question, as newly diagnosed patients exhibit considerable loss of rods and cones in their peripheral retinas. We have generated and characterized a tamoxifen inducible Cre-loxP rescue allele, Pde6b(Stop), which allows us to temporally correct PDE6-deficiency. Whereas untreated mutants exhibit degeneration, activation of Cre-loxP recombination in early embryogenesis produced stable long-term rescue. Reversal at later time-points showed partial long-term or short-lived rescue. Our results suggest stable restoration of retinal function by gene therapy can be achieved if a sufficient number of rods are treated. Because patients are generally diagnosed after extensive loss of rods, the success of clinical trials may depend on identifying patients as early as possible to maximize the number of treatable rods.
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Usefulness of plasma YKL-40 in management of community-acquired pneumonia severity in patients.
Int J Mol Sci
PUBLISHED: 08-07-2013
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Plasma YKL-40 level has been reported as playing a significant role in community-acquired pneumonia (CAP). However, the correlation between plasma level of YKL-40 and the severity of CAP has not been reported. This study identifies the relationship between plasma level changes of the YKL-40 gene in adult patients hospitalized with CAP. The ELISA was used to measure the plasma YKL-40 level from 61 adult CAP patients before and after antibiotic treatment and from 60 healthy controls. The plasma YKL-40 levels were significantly increased in patients with CAP compared to normal controls. Moreover, the plasma concentration of YKL-40 correlated with the severity of CAP based on the pneumonia severity index (PSI) score (r = 0.630, p < 0.001), the CURB-65 (confusion, uremia, respiratory rate, BP, age 65 years) score (r = 0.640, p < 0.001), the Acute Physiology And Chronic Health Evaluation II (APACHE II) score (r = 0.539, p < 0.001) and length of hospital stay (r = 0.321, p = 0.011), respectively. In conclusion, plasma YKL-40 may play a role in the diagnosis and clinical assessment of CAP severity, which could potentially guide the development of treatment strategies.
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Increased concentrations of plasma growth arrest-specific 6 and its soluble tyrosine kinase receptor sAxl in Taiwanese women with pelvic inflammatory disease.
Clin. Chim. Acta
PUBLISHED: 07-09-2013
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To investigate the concentrations of plasma growth arrest-specific protein 6 (Gas6) and its soluble tyrosine kinase receptor sAxl in women with pelvic inflammatory disease (PID) and their association with clinical outcomes of PID.
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New vascular ring connectors in surgery for intramural haematoma of the abdominal aorta progressing to rupture.
Cardiovasc J Afr
PUBLISHED: 06-19-2013
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Aortic intramural haematoma is similar to classic aortic dissection, which causes a life-threatening medical condition, and immediate diagnosis and treatment are crucial. The optimal therapy for intramural haematoma of the abdominal aorta remains controversial. Conservative medical management is usually the first choice of treatment for uncomplicated cases. Surgical intervention is usually required for complicated intramural haematomas of the abdominal aorta, including conventional open repair and endovascular treatment with stent-grafts. A new vascular ring connector that achieves a quick, blood-sealed and sutureless anastomosis has been designed for aortic dissection. We herein report a case of intramural haematoma of the abdominal aorta, progressing to rupture on day 14 after onset, which had successful aortic repair with the new vascular ring connector. The new vascular ring connector could be an alternative method for the treatment of complicated intramural haematomas of the abdominal aorta.
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Advanced glycation end products in degenerative nucleus pulposus with diabetes.
J. Orthop. Res.
PUBLISHED: 06-04-2013
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Diabetes mellitus (DM) has been clinically proved as a risk factor of disc degeneration, and the accumulation of advanced glycation end products (AGEs) is known to be potentially involved in diabetes. The purpose of this study is to investigate the effect of AGEs in the degeneration process of diabetic nucleus pulposus (NP) in rats and humans. Diabetic NP cells from rat coccygeal discs were treated with different concentrations of AGEs (0, 50, and 100?µg/ml) for 3 days, and mRNA expressions of MMP-2 and RAGE were measured by real-time RT-PCR. In addition, conditioned medium from NP cells was used to analyze protein expression of MMP-2 activity and ERK by gelatin zymography and Western blot. These experiments were repeated using human intervertebral disc samples. The immunohistochemical expression of AGEs was significantly increased in diabetic discs. In response to AGEs, an increase of MMP-2, RAGE, and ERK at both mRNA and protein expression levels was observed in diabetic NP cells. The findings suggest that AGEs and DM are associated with disc degeneration in both species. Hyperglycemia in diabetes enhances the accumulation of AGEs in the NP and triggers disc degeneration. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.
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Association between visfatin levels and coronary artery disease in patients with chronic kidney disease.
Iran J Kidney Dis
PUBLISHED: 05-26-2013
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Visfatin (also known as pre-B cell colony-enhancing factor) is increased in patients with chronic kidney disease and has been linked with coronary atherosclerosis. Given that it has been reported that visfatin plays a role in endothelial dysfunction in chronic kidney disease patients, we examined associations between visfatin levels and several markers related to atherosclerosis.
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Pb(2+) induced IL-8 gene expression by extracellular signal-regulated kinases and the transcription factor, activator protein 1, in human gastric carcinoma cells.
Environ. Toxicol.
PUBLISHED: 05-21-2013
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Divalent lead (Pb(2+) ) is a common industrial pollutant epidemiologically associated with gastric cancers. Pb(2+) was found to promote tumorigenesis, which may include interleukin (IL)-8, a pro-inflammatory chemokine that promotes angiogenesis and tumor metastasis. Given that the gastrointestinal tract is a major route of Pb(2+) exposure, we investigated the ability of Pb(2+) to induce IL-8 expression in gastric carcinoma cells and its underlying mechanism. At a concentration of 0.1 ?M, Pb(2+) induced IL-8 gene activation in gastric carcinoma AGS cells. Using a IL-8 promoter-deletion analysis, transcription factor activator protein 1 (AP-1) was identified as a necessary component of Pb(2+) -induced IL-8 gene activation. Upregulation of the IL-8 gene was abrogated by the MEK inhibitor, PD98059, and partially suppressed by the epidermal growth factor receptor inhibitors, AG1478 and PD153035. Furthermore, c-Jun protein expression was induced in cells treated with Pb(2+) , and overexpression of c-Jun enhanced Pb(2+) -induced IL-8 activation. Collectively, our findings highlight the pivotal roles of AP-1 and extracellular signal-regulated kinase in signal transduction of Pb(2+) -induced IL-8 gene activation. These molecules may be potential therapeutic targets for Pb(2+) -related inflammation leading to stomach carcinogenesis. © 2013 Wiley Periodicals, Inc. Environ Toxicol, 2013.
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A retrospective study of treating thoracolumbar spine fractures in ankylosing spondylitis.
Eur J Orthop Surg Traumatol
PUBLISHED: 04-29-2013
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Spinal fractures are commonly encountered in ankylosing spondylitis (AS) patients. This study compares the outcome of early surgical treatment with initial conservative treatment for thoracolumbar fractures in patients with AS. From 1996 to 2008, 28 patients with AS were treated either operatively or conservatively for thoracolumbar fractures; however, only 25 patients met the inclusion criteria with a minimum follow-up of 2 years. For surgically treated patients, posterior spinal instrumentation was performed using a transpedicle screw system. Nonsurgically treated patients wore a fracture brace. The demographic data, diagnosis, mechanism of injury, and neurological status were recorded, and fracture healing was assessed radiographically. The mean age was 54.2 ± 13.8 years (range 30-80 years). Six patients (Group A) received surgical intervention within 1 month. All of these fractures healed, and two of five patients showed neurologic improvement after surgery. Eight patients (Group B) had fractures that were missed. The delay in diagnosis resulted in pseudoarthrosis in all cases, and progressive neurologic deficits were identified in four cases. Eleven patients (Group C) received conservative treatment with bracing. Fracture union was achieved in three cases, and pseudoarthrosis occurred in eight cases. Operative treatment can achieve solid fusion and improve the neurological status, while conservative treatment may result in pseudoarthrosis and progressive neurologic deficit. The results suggest that AS patients with unstable spinal fractures should receive early surgical management to prevent further sequelae.
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A soft coral natural product, 11-episinulariolide acetate, inhibits gene expression of cyclooxygenase-2 and interleukin-8 through attenuation of calcium signaling.
Molecules
PUBLISHED: 04-22-2013
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Epidermal growth factor receptor (EGFR) is overexpressed in many types of cancer cells. EGFR-mediated signaling involves inflammatory gene expression including cyclooxygenase (COX)-2 and interleukin (IL)-8, and is associated with cancer pathogenesis. In a search of phytochemicals with anti-inflammatory activity, the COX-2 and IL-8 inhibitory activities of some marine compounds were examined. After screening these compounds 11-episinulariolide acetate (1) from soft coral exhibited the most potent activity. Reverse-transcription PCR; western blotting; ELISA and luciferase assays were used to test the effect of compound 1 on EGF-stimulated expressions of COX-2 and IL-8 in A431 human epidermoid carcinoma cells. After exposure to 10 ?M of compound 1, expression levels of COX-2 and IL-8 were reduced. In addition; intracellular Ca²? increase and Ca²?-dependent transcription factor activation were blocked by compound 1. Thus, compound 1 can potentially serve as a lead compound for targeting Ca²? signaling-dependent inflammatory diseases.
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Amikacin-induced Fin Reduction is Mediated by Autophagy.
J Toxicol Pathol
PUBLISHED: 04-22-2013
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Despite its medical use, little is known about the mechanisms underlying amikacin-induced embryotoxicity, including fin reduction, in zebrafish. In this study, we examined the expression of well-known autophagy markers mTOR (target of rapamycin), atg10 (autophagy-related gene), atg12 and LC3 (mammalian homolog of Atg8) in amikacin-treated zebrafish embryos. Our results indicated that the mRNA expression level of atg12 in the amikacin-treated group was significantly increased by 1.5-fold (p<0.05) compared with the corresponding mock control group, while the expression levels of atg10 and mTOR were significantly decreased by 0.74-fold (p<0.05) and 0.58-fold (p<0.05), respectively. Western blot analysis revealed that LC3 protein expression was induced by amikacin. Taken together, these data suggest that amikacin-induced fin reduction is mediated by fin cell autophagy.
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The effect of erythropoietin on autologous stem cell-mediated bone regeneration.
Biomaterials
PUBLISHED: 04-16-2013
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Mesenchymal stem cells (MSCs) although used for bone tissue engineering are limited by the requirement of isolation and culture prior to transplantation. Our recent studies have shown that biomaterial implants can be engineered to facilitate the recruitment of MSCs. In this study, we explore the ability of these implants to direct the recruitment and the differentiation of MSCs in the setting of a bone defect. We initially determined that both stromal derived factor-1alpha (SDF-1?) and erythropoietin (Epo) prompted different degrees of MSC recruitment. Additionally, we found that Epo and bone morphogenetic protein-2 (BMP-2), but not SDF-1?, triggered the osteogenic differentiation of MSCs in vitro. We then investigated the possibility of directing autologous MSC-mediated bone regeneration using a murine calvaria model. Consistent with our in vitro observations, Epo-releasing scaffolds were found to be more potent in bridging the defect than BMP-2 loaded scaffolds, as determined by computed tomography (CT) scanning, fluorescent imaging and histological analyses. These results demonstrate the tremendous potential, directing the recruitment and differentiation of autologous MSCs has in the field of tissue regeneration.
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Intestinal CCL11 and eosinophilic inflammation is regulated by myeloid cell-specific RelA/p65 in mice.
J. Immunol.
PUBLISHED: 04-05-2013
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In inflammatory bowel diseases (IBDs), particularly ulcerative colitis, intestinal macrophages (M?s), eosinophils, and the eosinophil-selective chemokine CCL11, have been associated with disease pathogenesis. M?s, a source of CCL11, have been reported to be of a mixed classical (NF-?B-mediated) and alternatively activated (STAT-6-mediated) phenotype. The importance of NF-?B and STAT-6 pathways to the intestinal M?/CCL11 response and eosinophilic inflammation in the histopathology of experimental colitis is not yet understood. Our gene array analyses demonstrated elevated STAT-6- and NF-?B-dependent genes in pediatric ulcerative colitis colonic biopsies. Dextran sodium sulfate (DSS) exposure induced STAT-6 and NF-?B activation in mouse intestinal F4/80(+)CD11b(+)Ly6C(hi) (inflammatory) M?s. DSS-induced CCL11 expression, eosinophilic inflammation, and histopathology were attenuated in RelA/p65(?mye) mice, but not in the absence of STAT-6. Deletion of p65 in myeloid cells did not affect inflammatory M? recruitment or alter apoptosis, but did attenuate LPS-induced cytokine production (IL-6) and Ccl11 expression in purified F4/80(+)CD11b(+)Ly6C(hi) inflammatory M?s. Molecular and cellular analyses revealed a link between expression of calprotectin (S100a8/S100a9), Ccl11 expression, and eosinophil numbers in the DSS-treated colon. In vitro studies of bone marrow-derived M?s showed calprotectin-induced CCL11 production via a p65-dependent mechanism. Our results indicate that myeloid cell-specific NF-?B-dependent pathways play an unexpected role in CCL11 expression and maintenance of eosinophilic inflammation in experimental colitis. These data indicate that targeting myeloid cells and NF-?B-dependent pathways may be of therapeutic benefit for the treatment of eosinophilic inflammation and histopathology in IBD.
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An emerging role for the anti-inflammatory cytokine interleukin-10 in dengue virus infection.
J. Biomed. Sci.
PUBLISHED: 03-28-2013
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Infection with dengue virus (DENV) causes both mild dengue fever and severe dengue diseases, such as dengue hemorrhagic fever and dengue shock syndrome. The pathogenic mechanisms for DENV are complicated, involving viral cytotoxicity, immunopathogenesis, autoimmunity, and underlying host diseases. Viral load correlates with disease severity, while the antibody-dependent enhancement of infection largely determines the secondary effects of DENV infection. Epidemiological and experimental studies have revealed an association between the plasma levels of interleukin (IL)-10, which is the master anti-inflammatory cytokine, and disease severity in patients with DENV infection. Based on current knowledge of IL-10-mediated immune regulation during infection, researchers speculate an emerging role for IL-10 in clinical disease prognosis and dengue pathogenesis. However, the regulation of dengue pathogenesis has not been fully elucidated. This review article discusses the regulation and implications of IL-10 in DENV infection. For future strategies against DENV infection, manipulating IL-10 may be an effective antiviral treatment in addition to the development of a safe dengue vaccine.
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Pb(2+) induces gastrin gene expression by extracellular signal-regulated kinases 1/2 and transcription factor activator protein 1 in human gastric carcinoma cells.
Environ. Toxicol.
PUBLISHED: 03-28-2013
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Divalent lead ions (Pb(2+) ) are toxic environmental pollutants known to cause serious health problems in humans and animals. Absorption of Pb(2+) from air, water, and food takes place in the respiratory and digestive tracts. The ways in which absorbed Pb(2+) affects cell physiology are just beginning to be understood at the molecular level. Here, we used reverse transcription PCR and Western blotting to analyze cultures of human gastric carcinoma cells exposed to 10 ?M lead nitrate. We found that Pb(2+) induces gastrin hormone gene transcription and translation in a time-dependent manner. Promoter deletion analysis revealed that activator protein 1 (AP1) was necessary for gastrin gene transcription in cells exposed to Pb(2+) . MitogIen-activated protein kinase (MAPK)/ERK kinase inhibitor PD98059 suppressed the Pb(2+) -induced increase in messenger RNA. Epidermal growth factor receptor (EGFR) inhibitors AG1478 and PD153035 reduced both transcription and phosphorylation by extracellular signal-regulated kinase (ERK1/2). Cells exposed to Pb(2+) also increased production of c-Jun protein, a component of AP1, and over-expression of c-Jun enhanced activation of the gastrin promoter. In sum, the findings suggest the EGFR-ERK1/2-AP1 pathway mediates the effects of Pb(2+) on gastrin gene activity in cell culture. © 2013 Wiley Periodicals, Inc. Environ Toxicol, 2013.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.