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Find video protocols related to scientific articles indexed in Pubmed.
Fabrication of porous titanium scaffolds by stack sintering of microporous titanium spheres produced with centrifugal granulation technology.
Mater Sci Eng C Mater Biol Appl
PUBLISHED: 07-11-2014
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Microporosity plays a key role in bioactivity and osteoinductivity of a biomaterial scaffold. A simple new approach to fabricating load-bearing porous titanium (Ti) scaffolds with uniform porous structure, highly controllable pore size and excellent biocompatibility was developed in the present study. This method was based on stack sintering of microporous Ti spheres produced with centrifugal granulation of commercial Ti powders. Macropores (180.0-341.8 ?m) and micropores (6.1-11.8 ?m) of the scaffolds were dependent on the sizes of the Ti spheres and the Ti powders, respectively. The compressive strength of the scaffolds (83.4-108.9 MPa) was high enough for the repair of load-bearing bone defects. Besides, the abundant micropores occurred on the rough and convex surface of the Ti spheres in the scaffolds were more favorable for adsorption of serum proteins, and thus promoted the growth of mesenchymal stem cells (MSCs).
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Facile fabrication of magnetic carbon composites from hydrochar via simultaneous activation and magnetization for triclosan adsorption.
Environ. Sci. Technol.
PUBLISHED: 04-29-2014
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Advanced magnetic carbon composites with high specific surface area and high microporosity are required for both environmentally and agriculturally related applications. However, more research is needed for the development of a facile and highly efficient synthesis process. In the present work, a novel approach of simultaneous activation and magnetization is proposed for the fabrication of magnetic carbon composites via the thermal pyrolysis of hydrochar (i.e., a solid residue from a hydrothermal carbonization process) that has been pretreated with mixtures of ferric chloride (FeCl3) and zinc chloride (ZnCl2). The main objective of this study is the investigation of the variation of characteristics of magnetic carbon composites produced at various conditions, as well as triclosan (TCS) adsorption behavior on such composites. This presented simple one-step synthesis method has the following advantages: (a) the hydrochar is activated with high surface area and pore volume (up to 1351 m(2)/g and 0.549 cm(3)/g, respectively), (b) activation and magnetization are simultaneously achieved without further modification, (c) the magnetic particles (?-Fe2O3) are stable under an acidic medium (pH of 3.0 and 4.0), and (d) the products have the potential to remove TCS from aqueous solutions with a maximum adsorption capacity of 892.9 mg/g. The results indicate the effectiveness of this facile synthesis strategy in converting low-value biowaste into a functional material with high performance for pollutant removal from aqueous solutions.
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Effect of phase composition on protein adsorption and osteoinduction of porous calcium phosphate ceramics in mice.
J Biomed Mater Res A
PUBLISHED: 01-22-2014
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The purpose of this study was to investigate the effect of phase compositions of porous calcium phosphate (CaP) ceramics on their protein adsorption behaviors in vitro and osteoinductive potentials in vivo in mice. Under competitive conditions, a high adsorption of bone morphogenetic protein 2 (BMP-2) was observed at a high initial concentration of BMP-2 in the multi-protein solution on all the four types of ceramics, indicating their strong affinity for BMP-2. No significant difference in BMP-2 adsorption between the ceramics was noted, indicating that phase composition could have little influence on BMP-2 adsorption. After implantation into the thigh muscles of mice for 45 and 90 days, the histological and histomorphometric analyses showed that porous biphasic calcium phosphate (BCP) ceramic consisting of 30% hydroxyapatite HA and 70% tricalcium phosphate (?-TCP), i.e. BCP-2 had stronger osteoinductive ability than the other three groups of ceramics. The immunohistochemical staining showed the highest expression of BMP-2 and osteocalcin (OCN) in BCP-2 group. Osteoinduction of porous CaP ceramics might be influenced by the amount of BMP-2 present in the local microenvironment in the implant, which was regulated by the phase composition of the ceramics. BCP-2 promoted the highest expression of BMP-2 and then showed the strongest osteoinduction in mice. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 4234-4243, 2014.
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Preparation of magnetic porous carbon from waste hydrochar by simultaneous activation and magnetization for tetracycline removal.
Bioresour. Technol.
PUBLISHED: 01-08-2014
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In the present work, a novel magnetic porous carbon (MPC) with maghemite (?-Fe2O3) particles is facilely prepared from hydrochar (a solid residue of hydrothermal carbonization of biomass) in one step through simultaneous activation and magnetization. The resultant MPC is characterized and utilized as an adsorbent for tetracycline (TC) removal from aqueous solutions. The BET surface area and micropore volume of the MPC are found to be 349 m(2)g(-1) and 0.16 cm(3)g(-1), respectively. The adsorption kinetics data could be well described by the pseudo-second-order model, and the TC adsorption onto MPC is an endothermic and spontaneous process. The enhanced surface area of the MPC, as well as its graphite-like structure, may contribute to the adsorption capacity of TC. After adsorption, MPC could be effectively separated by applying a magnetic field.
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TAT-Protein blockade during ischemia/reperfusion reveals critical role for p85 PI3K-PTEN interaction in cardiomyocyte injury.
PLoS ONE
PUBLISHED: 01-01-2014
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Recent work shows that cooling protection after mouse cardiac arrest and cardiomyocyte ischemia is mediated by Akt activation. The PI3K p85 subunit can either augment or inhibit Akt activation depending on its binding to p110 or PTEN respectively. To further clarify the role of PI3K p85 in cardioprotection, we studied novel TAT-p85 fusion proteins that selectively inhibit PI3K p85 binding. We hypothesized that TAT fused p85 lacking the PTEN binding site (TAT-?PTEN p85) would enhance Akt phosphorylation to afford cardioprotection. Conversely, TAT fused p85 lacking the p110 binding site (TAT-?p110p85) would decrease Akt phosphorylation and abrogate cardioprotection. Microscopy and Western blot analysis demonstrated that TAT fusion protein was transduced into cardiomyocytes within 5 min and remained more than 2 h. Inhibition of PI3K/Akt by TAT-?p110 p85 significantly increased cell death from 44.6±2.7% to 92.5±3.4% after simulated ischemia and reperfusion. By contrast, PTEN inhibition using TAT-?PTEN p85 decreased cell death to 11.9±5.3%, a similar level of cardioprotection seen with past cooling studies. Additional studies with the small molecule PTEN inhibitor VO-OHpic confirmed that PTEN inhibition was highly protective against cell death induced by ischemia and reperfusion. We conclude that blockade of p85-PTEN interaction and PTEN inhibition may be promising strategies for rescuing the heart from ischemia and reperfusion injury.
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VDR Attenuates Acute Lung Injury by Blocking Ang-2-Tie-2 Pathway and Renin-Angiotensin System.
Mol. Endocrinol.
PUBLISHED: 11-06-2013
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Acute lung injury (ALI) is a hallmark of systemic inflammation associated with high mortality. Although the vitamin D receptor (VDR) is highly expressed in the lung, its role in lung physiology remains unclear. We investigated the effect of VDR deletion on ALI using a lipopolysaccharide (LPS)-induced sepsis model. After LPS challenge VDR-null mice exhibited more severe ALI and higher mortality compared with wild-type (WT) counterparts, manifested by increased pulmonary vascular leakiness, pulmonary edema, apoptosis, neutrophil infiltration, and pulmonary inflammation, which was accompanied by excessive induction of angiopoietin (Ang)-2 and myosin light chain (MLC) phosphorylation in the lung. 1,25-Dihydroxyvitamin D blocked LPS-induced Ang-2 expression by blocking nuclear factor-?B activation in human pulmonary artery endothelial cells. The severity of lung injury seen in VDR-null mice was ameliorated by pretreatment with L1-10, an antagonist of Ang-2, suggesting that VDR signaling protects the pulmonary vascular barrier by targeting the Ang-2-Tie-2-MLC kinase cascade. Severe ALI in VDR-null mice was also accompanied by an increase in pulmonary renin and angiotensin II levels, and pretreatment of VDR-null mice with angiotensin II type 1 receptor blocker losartan partially ameliorated the severity of LPS-induced lung injury. Taken together, these observations provide evidence that the vitamin D-VDR signaling prevents lung injury by blocking the Ang-2-Tie-2-MLC kinase cascade and the renin-angiotensin system.
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Comparative assessment of the methanogenic steps of single and two-stage processes without or with a previous hydrolysis of cassava distillage.
Bioresour. Technol.
PUBLISHED: 06-26-2013
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In this study, cassava distillage with a high solid content was digested in an anaerobic sequencing batch reactor (ASBR) without or with a previous hydrolytic step by a cellulolytic microbial consortium (i.e., single or two-stage process). The methanogenic steps of these processes were compared and evaluated through observation of the methanogenic stability and methane yield under different organic loading rates (OLRs). It was found the methanogenic reactor can be stably performed with the OLRs lower than 20 g COD L(-1) d(-1) in the two-stage process, where a specific methane yield (0.147 L CH4 g(-1) CODremoved) could be achieved, which was 17.6% higher than that of the single-stage process (0.125 L CH4 g(-1) CODremoved). The above results indicated that the degradation of cassava distillage in a two-stage process with a previous hydrolytic step can assure the methanogenic process proceeds with greater stability and generates higher methane yield.
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Discovering small molecule ligands of vascular endothelial growth factor that block VEGF-KDR binding using label-free microarray-based assays.
Assay Drug Dev Technol
PUBLISHED: 06-19-2013
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We present here a label-free microarray-based assay platform that we used to identify inhibitors of vascular endothelial growth factor (VEGF)-kinase-insertion domain receptor (KDR) binding. Supported by a combination of special ellipsometry-based optical detection and small molecule microarrays (SMM), this platform consists of three assays: (1) the first assay detects binding of a target protein with SMM and identifies ligands to the protein as inhibitor candidates; (2) the second assay detects binding of a receptor protein with identical SMM and subsequent binding of the target protein (a sandwich assay) to identify the ligands to the receptor protein that do not interfere with the target-receptor binding; (3) the third assay detects binding of the target protein to the receptor protein in the presence of the ligands of the target protein identified from the first assay, with the receptor protein immobilized to a solid surface through the ligands identified in the second assay, to yield dose-response curves. Using this platform, we screened 7,961 compounds from the National Cancer Institute and found 12 inhibitors to VEGF-KDR (VEGFR2) interactions with IC?? ranging from 0.3 to 60 ?M. The inhibitory potency of these inhibitors found in the microarray-based assay was confirmed by their inhibition of VEGF-induced VEGFR2 phosphorylation in a cell-based assay.
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Isolation of new flavan-3-ol and lignan glucoside from Loropetalum chinense and their antimicrobial activities.
Fitoterapia
PUBLISHED: 05-12-2013
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Phytochemical and antimicrobial activity study on the ethanol extract of the leaves and stems of Loropetalum chinense led to the isolation of a new flavan-3-ol compounds, 8-[1-(3,4-dihydroxyphenyl)-3-methoxy-3-oxopropyl]-catechin (loropetaliside A) (1) and a new lignan glucoside, 1-(5-hydroxy-3-methoxyphenyl)-2-(2-?-glucopyranosyl-4-hydroxy-5-(1-(E)propen-3-ol)-phenyl)-propane-3-ol (loropetaliside B) (3) and several known compounds manglieside D (2), quercetin (4), kaempferol-3-O-D-glucopyranoside (5), quercetin-3-O-?-L-rhamnoside (6) and tiliroside (7). Their structures were elucidated on the basis of extensive spectroscopic analysis.
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Baicalein preconditioning protects cardiomyocytes from ischemia-reperfusion injury via mitochondrial oxidant signaling.
Am. J. Chin. Med.
PUBLISHED: 04-04-2013
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Previous studies suggest baicalein, in addition to its antioxidant effects, protects against hypoxia/reoxygenation injury via its pro-oxidant properties. We hypothesize that a brief period of baicalein treatment prior to ischemia/reperfusion (I/R) may trigger preconditioning protection via a mitochondrial pro-oxidant mechanism. Using an established chick cardiomyocyte model of I/R, cells were preconditioned with baicalein (10 ?M) for 10 min followed by 10-min wash prior to I/R. Intracellular oxidants were measured using 2, 7-dichlorofluorescin diacetate (DCFH/DA). Cell viability was assessed by propidium iodide and apoptosis determined by DNA fragmentation. Baicalein induced a transient but significant increase of DCF fluorescence within the 10-min preconditioning period, and led to significant reduction of cell death (38.9 ± 1.8% vs. 58.7 ± 1.2% in I/R control, n = 6, p < 0.001) and DNA fragmentation after I/R. Cotreatment with N-acetylcysteine (500 ?M), mitochondrial complex III electron transport chain inhibitor myxothiazol (1 ?M), mitochondrial KATP channel blocker 5-hydroxydecanoate-Na (5-HD, 500 ?M) or anion channel inhibitor 4, 4-diisothiocyanato-stilbene-2, 2-disulfonic acid (DIDS, 200 ?M) resulted in significant abrogation of oxidant increase during induction as well as the protection conferred by baicalein preconditioning. These results suggest that baicalein preconditioning exhibits significant anti-apoptotic protection against cardiomyocyte I/R injury by mitochondrial oxidant signaling, which was in part mediated by mitochondrial KATP channel and anion channel opening.
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Dynamic competitive adsorption of bone-related proteins on calcium phosphate ceramic particles with different phase composition and microstructure.
J. Biomed. Mater. Res. Part B Appl. Biomater.
PUBLISHED: 01-12-2013
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The biocompatibility and bioactivity of biomaterials used for hard tissue repair are closely related to their adsorption capacities for bone-related proteins. In the present study, three types of calcium phosphate (CaP) ceramic particles with different phase composition or microstructure were fabricated, and their protein adsorption abilities were investigated by a self-made device under the simulated dynamic physiological circumstance. The results of X-ray diffraction, field emission scanning electron microscopy, mercury penetration test, and nitrogen sorption test showed that the irregular hydroxyapatite (HA) ceramic particles obtained by conventional drying and sintering (named as HA-C) had fewer micropores and lower specific surface area (SSA) than did the spherical HA or biphasic calcium phosphate (BCP) ceramic particles made by spray drying and sintering (named as HA-S and BCP-S, respectively). The dynamic protein adsorption study proved that both the phase composition and microstructure of CaP ceramic particles affected their adsorption capacities for those bone-related proteins. The spherical HA-S and BCP-S particles with abundant micropores and high SSA showed higher adsorption of serum proteins, including fibronectin and vitronectin, than the irregular HA-C did. On the other hand, in spite of the relatively high concentration of bovine serum albumin (BSA) in the binary bone morphogenetic protein 2 (BMP-2)/BSA solution, BMP-2 adsorption on the three CaP ceramic particles increased with the increase in its initial concentration. Similarly, HA-S and BCP-S particles had a larger amount of the adsorbed BMP-2 per gram solid than HA-C did. Therefore, it could be believed that the difference of various CaP ceramics in the phase composition and microporous structure would affect their binding capacity for those bone-related proteins and thus lead to their difference in osteoinduction.
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Repair of large osteochondral defects in a beagle model with a novel type I collagen/glycosaminoglycan-porous titanium biphasic scaffold.
Mater Sci Eng C Mater Biol Appl
PUBLISHED: 01-10-2013
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The limited repair potential of articular cartilage, which hardly heals after injury or debilitating osteoarthritis, is a clinical challenge. The aim of this work was to develop a novel type I collagen (Col)/glycosaminoglycan (GAGs)-porous titanium biphasic scaffold (CGT) and verify its ability to repair osteochondral defects in an animal model with bone marrow stem cells (bMSCs) in the chondral phase. The biphasic scaffold was composed of Col/GAGs as chondral phasic and porous titanium as subchondral phasic. Twenty-four full-thickness defects through the articular cartilage and into the subchondral bone were prepared by drilling into the surface of the femoral patellar groove. Animals were assigned to one of the three groups: 1) CGT with bMSCs (CGTM), 2) only CGT, and 3) no implantation (control). The defect areas were examined grossly, histologically and by micro-CT. The most satisfied cartilage repairing result was in the CGTM group, while CGT alone was better than the control group. Abundant subchondral bone formation was observed in the CGTM and CGT groups but not the control group. Our findings demonstrate that a composite based on a novel biphasic scaffold combined with bMSCs shows a high potential to repair large osteochondral defects in a canine model.
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Oleanolic acid and ursolic acid: novel hepatitis C virus antivirals that inhibit NS5B activity.
Antiviral Res.
PUBLISHED: 01-10-2013
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Hepatitis C virus (HCV) infects up to 170 million people worldwide and causes significant morbidity and mortality. Unfortunately, current therapy is only curative in approximately 50% of HCV patients and has adverse side effects, which warrants the need to develop novel and effective antivirals against HCV. We have previously reported that the Chinese herb Fructus Ligustri Lucidi (FLL) directly inhibited HCV NS5B RNA-dependent RNA polymerase (RdRp) activity (Kong et al., 2007). In this study, we found that the FLL aqueous extract strongly suppressed HCV replication. Further high-performance liquid chromatography (HPLC) analysis combined with inhibitory assays indicates that oleanolic acid and ursolic acid are two antiviral components within FLL aqueous extract that significantly suppressed the replication of HCV genotype 1b replicon and HCV genotype 2a JFH1 virus. Moreover, oleanolic acid and ursolic acid exhibited anti-HCV activity at least partly through suppressing HCV NS5B RdRp activity as noncompetitive inhibitors. Therefore, our results for the first time demonstrated that natural products oleanolic acid and ursolic acid could be used as potential HCV antivirals that can be applied to clinic trials either as monotherapy or in combination with other HCV antivirals.
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Simultaneous measurement of 10,000 protein-ligand affinity constants using microarray-based kinetic constant assays.
Assay Drug Dev Technol
PUBLISHED: 12-22-2011
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Fluorescence-based endpoint detection of microarrays with 10,000 or more molecular targets is a most useful tool for high-throughput profiling of biomolecular interactions, including screening large molecular libraries for novel protein ligands. However, endpoint fluorescence data such as images of reacted microarrays contain little information on kinetic rate constants, and the reliability of endpoint data as measures of binding affinity depends on reaction conditions and postreaction processing. We here report a simultaneous measurement of binding curves of a protein probe with 10,000 molecular targets in a microarray with an ellipsometry-based (label-free) optical scanner. The reaction rate constants extracted from these curves (k(on), k(off), and k(a)=k(on)/k(off)) are used to characterize the probe-target interactions instead of the endpoints. This work advances the microarray technology to a new milestone, namely, from an endpoint assay to a kinetic constant assay platform. The throughput of this binding curve assay platform is comparable to those at the National Institutes of Health Molecular Library Screening Centers, making it a practical method in screening compound libraries for novel ligands and for system-wide affinity profiling of proteins, viruses, or whole cells against diverse molecular targets.
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Humic acid and metal ions accelerating the dechlorination of 4-chlorobiphenyl by nanoscale zero-valent iron.
J Environ Sci (China)
PUBLISHED: 12-02-2011
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Transformation of polychlorinated biphenyls (PCBs) by zero-valent iron represents one of the latest innovative technologies for environmental remediation. The dechlorination of 4-chlorobiphenyl (4-ClBP) by nanoscale zero-valent iron (NZVI) in the presence of humic acid or metal ions was investigated. The results showed that the dechlorination of 4-ClBP by NZVI increased with decreased solution pH. When the initial pH value was 4.0, 5.5, 6.8, and 9.0, the dechlorination efficiencies of 4-ClBP after 48 hr were 53.8%, 47.8%, 35.7%, and 35.6%, respectively. The presence of humic acid inhibited the reduction of 4-ClBP in the first 4 hr, and then significantly accelerated the dechlorination by reaching 86.3% in 48 hr. Divalent metal ions, Co2+, Cu2+, and Ni2+, were reduced and formed bimetals with NZVI, thereby enhanced the dechlorination of 4-ClBP. The dechlorination percentages of 4-ClBP in the presence of 0.1 mmol/L Co2+, Cu2+ and Ni2+ were 66.1%, 66.0% and 64.6% in 48 hr, and then increased to 67.9%, 71.3% and 73.5%, after 96 hr respectively. The dechlorination kinetics of 4-ClBP by the NZVI in all cases followed pseudo-first order model. The results provide a basis for better understanding of the dechlorination mechanisms of PCBs in real environment.
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Label-free detection of surface markers on stem cells by oblique-incidence reflectivity difference microscopy.
BioTechniques
PUBLISHED: 11-06-2011
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Conventional fluorescence microscopy is routinely used to detect cell surface markers through fluorophore-conjugated antibodies. However, fluorophore-conjugation of antibodies alters binding properties such as strength and specificity of the antibody in often uncharacterized ways. Here we present a method using an oblique-incidence reflectivity difference (OI-RD) microscope for label-free, real-time detection of cell surface markers, and apply it to analysis of stage-specific embryonic antigen 1 (SSEA1) on stem cells. Mouse stem cells express SSEA1 on their surfaces, and the level of SSEA1 decreases when the cells start to differentiate. In this study, we immobilized mouse stem cells and non-stem cells (control) on a glass surface as a microarray and reacted the cell microarray with unlabeled SSEA1 antibodies. By monitoring the reaction with an OI-RD microscope in real time, we confirmed that the SSEA1 antibodies bind only to the surface of the stem cells and not to the surface of non-stem cells. From the binding curves, we determined the equilibrium dissociation constant (Kd) of the antibody with the SSEA1 markers on the stem cell surface. Thus, the OI-RD microscope can be used to detect binding affinities between cell surface markers and unlabeled antibodies bound to the cells; this information could be useful for determination of stem cell stages.
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Fluorescent labeling agents change binding profiles of glycan-binding proteins.
Mol Biosyst
PUBLISHED: 10-18-2011
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Interactions of glycan-binding proteins (GBPs) with glycans are essential in cell adhesion, bacterial/viral infection, and cellular signaling pathways. Experimental characterization of these interactions based on glycan microarrays typically involves (1) labeling GBPs directly with fluorescent reagents before incubation with the microarrays, or (2) labeling GBPs with biotin before the incubation and detecting the captured GBPs after the incubation using fluorescently labeled streptavidin, or (3) detecting the captured GBPs after the incubation using fluorescently labeled antibodies raised against the GBPs. The fluorescent signal is mostly measured ex situ after excess fluorescent materials are washed off. In this study, by using a label-free optical scanner for glycan microarray detection, we measured binding curves of 7 plant lectins to 24 glycans: four ?1-4-linked galactosides, three ?1-3-linked galactosides, one ?-linked galactoside, one ?-linked N-acetylgalactosaminide, eight ?2-3-linked sialosides, and seven ?2-6-linked sialosides. From association and dissociation constants deduced by global-fitting the binding curves, we found that (1) labeling lectins directly with fluorescent agents change binding profiles of lectins, in some cases by orders of magnitude; (2) those lectin-glycan binding reactions characterized with large dissociation rates, though biologically relevant, are easily missed or deemed insignificant in ex situ fluorescence-based assays as most captured lectins are washed off before detection. This study highlights the importance of label-free real-time detection of protein-ligand interactions and the potential pitfall in interpreting fluorescence-based assays for characterization of protein-glycan interactions.
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Delayed intracerebral hemorrhage secondary to ventriculoperitoneal shunt: two case reports and a literature review.
Int J Med Sci
PUBLISHED: 09-21-2011
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Ventriculoperitoneal (VP) shunt has become a popular operation to achieve cerebrospinal fluid (CSF) diversion, but is associated with many complications. Postoperative delayed intracerebral hemorrhage is a kind of rare but severe event, which has not thus far been reported in retrospective case analyses. Here we present two cases of delayed intracerebral hemorrhage, along the path of the ventricular catheter, which occurred on postoperative days 3 and 5. We also provide a literature review regarding this rare complication.
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A case of McCune-Albright syndrome associated with pituitary GH adenoma: therapeutic process and autopsy.
J. Pediatr. Endocrinol. Metab.
PUBLISHED: 08-10-2011
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McCune-Albright syndrome (MAS) is a clinical syndrome with low incidence, and its concurrence with pituitary GH adenoma is rare. Little of the history, treatment and outcome has been studied.
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New pregnane saponins from Ecdysanthera rosea and their cytotoxicity.
Fitoterapia
PUBLISHED: 01-25-2011
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Two new pregnane saponins elucidated as ecdysantheroside A (1) and ecdysantheroside B (2) and six known compounds (3-8) based on spectral data (MS, IR, 1D and 2D NMR) were isolated from the stem bark of Ecdysanthera rosea. The cytotoxicity against six cell lines of these compounds was tested by MTT assay. The results revealed that compounds 5 and 7 showed cytotoxicity against all the cell lines. Compound 2 showed cytotoxicity against cells A549, MDA435, HepG2, and HUVEC, while compound 4 showed cytotoxicity against cells A549, CEM, and HUVEC. Compound 6 had cytotoxicity against the others except cell HepG2.
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Subcellular Cd distribution and its correlation with antioxidant enzymatic activities in wheat (Triticum aestivum) roots.
Ecotoxicol. Environ. Saf.
PUBLISHED: 10-13-2010
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The subcellular partitioning of Cd in wheat root was obtained by differential centrifugation of root homogenates to quantify the amounts of Cd associated with five operationally defined subcellular fractions, namely Cd-rich granule (MRG), cellular debris, organelles, heat-denatured protein (HDP), and heat-stable protein (HSP). Their correlations were investigated with the changes in antioxidant enzymatic activities under Cd stress (0.01-9.68 ?M) for 72 h. The results showed that the relative Cd distribution in detoxified fractions (MRG+HSP) was decreased with increasing [Cd], but in metal-sensitive fractions (HDP+organelles) was increased. Consequently, the content of superoxide dismutase (SOD) and catalase (CAT) was decreased. Additionally, an increase of the total antioxidant capacity (A-TOC) and a slight decrease of malondialdehyde (MDA) were also observed. Interestingly, the relative root elongation (RRL) and CAT were significantly correlated with the Cd-sensitive fractions. These results suggest that the Cd toxicity to wheat roots strongly depends on the subcellular Cd distribution.
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Osteoinduction of porous titanium: a comparative study between acid-alkali and chemical-thermal treatments.
J. Biomed. Mater. Res. Part B Appl. Biomater.
PUBLISHED: 09-30-2010
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In this study, a slurry foaming method was developed to fabricate porous titanium, and two different surface treatments were applied to investigate their effects on the osteoinduction of the implants. Three types of implants, that was porous titanium with no treatment, with chemical-thermal treatment (CTPT), and with acid-alkali treatment (AAPT), were implanted in the dorsal muscles of adult dogs for 3 and 5 months. After implantation for 3 months, new bone was only found in the inner pores of AAPT by histological analysis and field emission scanning electron microscopy observation. After implantation for 5 months, new bone was also found in CTPT, but it was absent in AAPT. This study not only confirmed that porous titanium with appropriate surface treatments could possess osteoinduction but also showed that its osteoinductive potential was tightly related to the surface treatment. As a simpler method, acid-alkali treatment could endow porous titanium with faster osteoinduction, and AAPT might have potential in clinical application.
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Development of novel microsatellite DNA markers by cross-amplification and analysis of genetic variation in gerbils.
J. Hered.
PUBLISHED: 06-04-2010
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The objectives of this study are to establish microsatellite loci for the Mongolian gerbil based on mouse microsatellite DNA sequences and to investigate genetic variation in the laboratory gerbil (Capital Medical University, CMU) and 2 wild gerbil populations (from Yin Chuan city [YIN] and the Hohehot Municipality [HOH]). In total, 536 mouse microsatellite markers were chosen to identify polymorphic dinucleotide repeat loci in the gerbil by cross-amplification. Of these markers, 313 (58.39%) have been discretely amplified from the CMU laboratory gerbil and been sequenced. Of the 313 sequenced markers, 130 were confirmed as simple sequence repeat (SSR) loci in the gerbil. In total, 6 of those newly identified loci plus 6 identified in previous reports were used to estimate the genetic polymorphism for 30 laboratory gerbils and 54 wild gerbils (27 each of the HOH and YIN groups). A total of 29 alleles were observed in the 3 populations, and 11 of 12 loci (91.67%) are polymorphic markers. Neis standard genetic distances of 0.0592 (CMU vs. HOH) and 0.1033 (CMU vs. YIN) were observed. The averages of observed versus expected heterozygosity are 0.5231/0.4008, 0.5051/0.3882, and 0.4825/0.3665 for the YIN, HOH, and CMU populations, respectively. These results show that cross-amplification using mouse microsatellite primers is an efficient way to identify gerbil SSR loci. By using these 12 selected markers, we have demonstrated that genetic variation level within the CMU population is higher than that has been reported previously and are comparable with the levels found in 2 wild populations.
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New hydroquinone diglycoside acyl esters and sesquiterpene and apocarotenoid from Ecdysanthera rosea.
Fitoterapia
PUBLISHED: 05-28-2010
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Phytochemical studies on the ethanol extract of the aerial parts of Ecdysanthera rosea led to the isolation of three new compounds, hydroquinone diglycoside acyl esters, ecdysanrosin A (1) and sesquiterpene, 5?-hydroperoxycostic acid (2) and apocarotenoid, 2, 4, 7-trimethyl-2, 4, 6, 8-tetraene-dialdehyde (3). Their structures were elucidated on the basis of extensive spectroscopic analysis.
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Cytosolic group IVa phospholipase A2 mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro.
J Inflamm (Lond)
PUBLISHED: 03-18-2010
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Cytosolic gIVaPLA2 is a critical enzyme in the generation of arachidonate metabolites and in induction of beta2-integrin adhesion in granulocytes. We hypothesized that gIVaPLA2 activation also is an essential downstream step for post adhesive migration of PMN in vitro.
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Screening small-molecule compound microarrays for protein ligands without fluorescence labeling with a high-throughput scanning microscope.
J Biomed Opt
PUBLISHED: 03-10-2010
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We describe a high-throughput scanning optical microscope for detecting small-molecule compound microarrays on functionalized glass slides. It is based on measurements of oblique-incidence reflectivity difference and employs a combination of a y-scan galvometer mirror and an x-scan translation stage with an effective field of view of 2 cm x 4 cm. Such a field of view can accommodate a printed small-molecule compound microarray with as many as 10,000 to 20,000 targets. The scanning microscope is capable of measuring kinetics as well as endpoints of protein-ligand reactions simultaneously. We present the experimental results on solution-phase protein reactions with small-molecule compound microarrays synthesized from one-bead, one-compound combinatorial chemistry and immobilized on a streptavidin-functionalized glass slide.
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Hematopoietic Pyk2 regulates migration of differentiated HL-60 cells.
J Inflamm (Lond)
PUBLISHED: 02-09-2010
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Pyk2 is a non-receptor cytoplasmic tyrosine kinase that belongs to the focal adhesion kinase family and has been implicated in neutrophil spreading and respiratory burst activity caused by TNF-alpha. However, the role of Pyk2 in neutrophil migration is incompletely defined. In this study, we tested the hypothesis that Pyk2 regulates the migration of neutrophil-like differentiated HL-60 cells subsequent to beta2-integrin mediated cell adhesion.
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Development of a noise metric for assessment of exposure risk to complex noises.
J. Acoust. Soc. Am.
PUBLISHED: 07-31-2009
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Many noise guidelines currently use A-weighted equivalent sound pressure level L(Aeq) as the noise metric and the equal energy hypothesis to assess the risk of occupational noises. Because of the time-averaging effect involved with the procedure, the current guidelines may significantly underestimate the risk associated with complex noises. This study develops and evaluates several new noise metrics for more accurate assessment of exposure risks to complex and impulsive noises. The analytic wavelet transform was used to obtain time-frequency characteristics of the noise. 6 basic, unique metric forms that reflect the time-frequency characteristics were developed, from which 14 noise metrics were derived. The noise metrics were evaluated utilizing existing animal test data that were obtained by exposing 23 groups of chinchillas to, respectively, different types of noise. Correlations of the metrics with the hearing losses observed in chinchillas were compared and the most promising noise metric was identified.
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Inhibition of Pyk2 blocks airway inflammation and hyperresponsiveness in a mouse model of asthma.
Am. J. Respir. Cell Mol. Biol.
PUBLISHED: 06-11-2009
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The objective of this investigation was to determine the role of Pyk2, an intracellular nonreceptor protein tyrosine kinase for postadhesive inflammatory cell migration, on airway inflammation and hyperresponsiveness in immune-sensitized mice. Blockade of Pyk2 was effected by intraperitoneal administration of dominant-negative C-terminal Pyk2 fused to a TAT protein transduction domain (TAT-Pyk2-CT). Ovalbumin challenge elicited infiltration of both eosinophils and lymphocytes into airways, increased mucus-containing epithelial cells, and caused increased airway hyperresponsiveness to methacholine in immune-sensitized mice. Pretreatment with 10 mg/kg TAT-Pyk2-CT intraperitoneally blocked all of these effects and further decreased secretion of Th2 cytokine IL-4, IL-5, and IL-13 into the bronchoalveolar lavage fluid. Intranasal administration of IL-5 caused eosinophil migration into the airway lumen, which was attenuated by systemic pretreatment with TAT-Pyk2-CT. In each paradigm, treatment with control protein TAT-GFP had no blocking effect. We conclude that Pyk2, which is essential for inflammatory cell migration in vitro, regulates airway inflammation, Th2 cytokine secretion, and airway hyperresponsiveness in the ovalbumin-sensitized mice during antigen challenge in vivo.
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Hepatitis C virus NS4B induces unfolded protein response and endoplasmic reticulum overload response-dependent NF-kappaB activation.
Virology
PUBLISHED: 04-14-2009
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Hepatitis C virus nonstructural protein 4B (NS4B) is an endoplasmic reticulum (ER) membrane associated protein and a potent causative factor of ER stress. Here we reported that unfolded protein response (UPR) can be activated by HCV NS4B through inducing both XBP1 mRNA splicing and ATF6 cleavage in human hepatic cells. Flow cytometric analysis revealed that HCV NS4B stimulates the production of reactive oxygen species (ROS) by perturbing intracellular Ca(2+) homeostasis. Luciferase assay showed that HCV NS4B also activates the multifunctional transcription factor, NF-kappaB, in a dose-dependent manner through Ca(2+) signaling and ROS. Further immunoblot analysis showed that HCV NS4B promotes NF-kappaB translocation into the nucleus via protein-tyrosine kinase (PTK) mediated phosphorylation and subsequent degradation of IkappaBalpha. These studies provide an important insight into the implication of NS4B in HCV life cycle and HCV-associated liver disease by affecting host intracellular signal transduction pathways.
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Addition of sodium hyaluronate and the effect on performance of the injectable calcium phosphate cement.
J Mater Sci Mater Med
PUBLISHED: 03-03-2009
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An injectable calcium phosphate cement (CPC) with porous structure and excellent anti-washout ability was developed in the study. Citric acid and sodium bicarbonate were added into the CPC powder consisting of tetracalcium phosphate (TTCP) and dicalcium phosphate dihydrate (DCPD) to form macro-pores, then different concentrations of sodium hyaluronate (NaHA) solution, as liquid phase, was added into the cement to investigate its effect on CPCs performance. The prepared CPCs were tested on workability (injectable time and setting time), mechanical strength, as well as anti-washout ability. The experimental results showed that addition of NaHA not only enhanced the anti-washout ability of the CPC dramatically but also improve its other properties. When NaHA concentration was 0.6 wt%, the injectable time elongated to 15.7 +/- 0.6 min, the initial and final setting times were respectively shorten to 18.3 +/- 1.2 and 58.7 +/- 2.1 min, and the compressive strength were increased to 18.78 +/- 1.83 MPa. On the other hand, Addition of NaHA showed little effect on porous structure of the CPC and enhanced its bioactivity obviously, which was confirmed by the apatite formation on its surface after immersion in simulated body fluid (SBF). In conclusion, as an in situ shaped injectable biomaterials, the CPC with appropriate addition of NaHA would notably improve its performance and might be used in minimal invasive surgery for bone repair or reconstruction.
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Effect of nanostructure on osteoinduction of porous biphasic calcium phosphate ceramics.
Acta Biomater
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In order to evaluate the effect of the nanostructure of calcium phosphate ceramics on osteoinductive potential, porous biphasic calcium phosphate (BCP) ceramics with a nano- or submicron structure were prepared via microwave sintering and compared to conventional BCP ceramics. The selective protein adsorption of bovine serum albumin and lysozyme (LSZ) and the osteogenic differentiation of human mesenchymal stem cells in vitro was investigated. Porous BCP nanoceramics showed higher ability to adsorb proteins, especially low molecular weight protein of LSZ, than conventional BCP ceramics, and the BCP nanoceramics promoted bone sialoprotein expression more than conventional BCP did. Further in vivo study to investigate ectopic bone formation and bone repair efficiency proved the highly osteoinductive potential of nanostructured BCP ceramics. The results suggest that nanostructured BCP ceramics have the potential to become a new generation of bioceramics for bone tissue engineering grafts.
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Etiologic diagnosis of lower respiratory tract bacterial infections using sputum samples and quantitative loop-mediated isothermal amplification.
PLoS ONE
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Etiologic diagnoses of lower respiratory tract infections (LRTI) have been relying primarily on bacterial cultures that often fail to return useful results in time. Although DNA-based assays are more sensitive than bacterial cultures in detecting pathogens, the molecular results are often inconsistent and challenged by doubts on false positives, such as those due to system- and environment-derived contaminations. Here we report a nationwide cohort study on 2986 suspected LRTI patients across P. R. China. We compared the performance of a DNA-based assay qLAMP (quantitative Loop-mediated isothermal AMPlification) with that of standard bacterial cultures in detecting a panel of eight common respiratory bacterial pathogens from sputum samples. Our qLAMP assay detects the panel of pathogens in 1047(69.28%) patients from 1533 qualified patients at the end. We found that the bacterial titer quantified based on qLAMP is a predictor of probability that the bacterium in the sample can be detected in culture assay. The relatedness of the two assays fits a logistic regression curve. We used a piecewise linear function to define breakpoints where latent pathogen abruptly change its competitive relationship with others in the panel. These breakpoints, where pathogens start to propagate abnormally, are used as cutoffs to eliminate the influence of contaminations from normal flora. With help of the cutoffs derived from statistical analysis, we are able to identify causative pathogens in 750 (48.92%) patients from qualified patients. In conclusion, qLAMP is a reliable method in quantifying bacterial titer. Despite the fact that there are always latent bacteria contaminated in sputum samples, we can identify causative pathogens based on cutoffs derived from statistical analysis of competitive relationship.
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Inhibition of Pyk2 blocks lung inflammation and injury in a mouse model of acute lung injury.
Respir. Res.
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Proline-rich tyrosine kinase 2 (Pyk2) is essential in neutrophil degranulation and chemotaxis in vitro. However, its effect on the process of lung inflammation and edema formation during LPS induced acute lung injury (ALI) remains unknown. The goal of the present study was to determine the effect of inhibiting Pyk2 on LPS-induced acute lung inflammation and injury in vivo.
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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.