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Find video protocols related to scientific articles indexed in Pubmed.
Delineation of the Role of Glycosylation in the Cytotoxic Properties of Quercetin using Novel Assays in Living Vertebrates.
J. Nat. Prod.
PUBLISHED: 11-15-2014
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Quercetin is a plant-derived flavonoid and its cytotoxic properties have been widely reported. However, in nature, quercetin predominantly occurs as various glycosides. Thus far the cytotoxic activity of these glycosides has not been investigated to the same extent as quercetin, especially in animal models. In this study, the cytotoxic properties of quercetin (1), hyperoside (quercetin 3-O-galactoside, 2), isoquercitrin (quercetin 3-O-glucoside, 3), quercitrin (quercetin 3-O-rhamnoside, 4), and spiraeoside (quercetin 4'-O-glucoside, 5) were directly compared in vitro using assays of cancer cell viability. To further characterize the influence of glycosylation in vivo, a novel zebrafish-based assay was developed that allows the rapid and experimentally convenient visualization of glycoside cleavage in the digestive tract. This assay was correlated with a novel human tumor xenograft assay in the same animal model. The results showed that 3 is as effective as 1 at inhibiting cancer cell proliferation in vivo. Moreover, it was observed that 3 can be effectively deglycosylated in the digestive tract. Collectively, these results indicate that 3 is a very promising drug candidate for cancer therapy, because glycosylation confers advantageous pharmacological changes compared with the aglycone, 1. Importantly, the development of a novel and convenient fluorescence-based assay for monitoring deglycosylation in living vertebrates provides a valuable platform for determining the metabolic fate of naturally occurring glycosides.
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Intake levels of dietary polyunsaturated fatty acids modify the association between the genetic variation in PCSK5 and HDL cholesterol.
J. Med. Genet.
PUBLISHED: 10-28-2014
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A low serum level of high-density lipoprotein cholesterol (HDL-C) is a risk factor for cardiovascular disease. Proprotein convertase subtilisin/kexin type 5 (PCSK5) modulates HDL-C metabolism through the inactivation of endothelial lipase activity.
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A Single Module Type I Polyketide Synthase Directs De Novo Macrolactone Biogenesis during Galbonolide Biosynthesis in Streptomyces galbus.
J. Biol. Chem.
PUBLISHED: 10-23-2014
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Galbonolide (GAL)-A and -B are antifungal macrolactone polyketides produced by Streptomyces galbus. During their polyketide chain assembly, GAL-A and -B incorporate methoxymalonate and methylmalonate, respectively, in the fourth chain extension step. The methoxymalonyl-acyl carrier protein (-ACP) biosynthesis locus (galG to K) is specifically involved in GAL-A biosynthesis, and this locus is neighbored by a gene cluster composed of galA-E. GalA-C constitute a single module, highly reducing type I polyketide synthase (PKS). GalD and GalE are cytochrome P450 and Rieske domain protein, respectively. Gene knock-out experiments verified that galB, C, and D are essential for GAL biosynthesis. A galD mutant accumulated a GAL-C that lacked two hydroxyl groups and a double bond when compared to GAL-B. A [U-13C] propionate feeding experiment indicated that no rare precursor, other than methoxymalonate, was incorporated during GAL biogenesis. A search of the S. galbus genome for a modular type I PKS system, the type that was expected to direct GAL biosynthesis, resulted in the identification of only one modular type I PKS gene cluster. Homology analysis indicated that this PKS gene cluster is the locus for vicenistatin biosynthesis. This cluster was previously reported in Streptomyces halstedii. A gene deletion of the vinP2 ortholog clearly demonstrated that this modular type I PKS system is not involved in GAL biosynthesis. Therefore, we propose that GalA-C direct macrolactone polyketide formation for GAL. Our studies provide a glimpse into a novel biochemical strategy used for polyketide synthesis: the iterative assembly of propionates with highly programmed ?-keto group modifications.
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New Platform for Cytochrome P450 Reaction Combining in Situ Immobilization on Biopolymer.
Bioconjug. Chem.
PUBLISHED: 10-17-2014
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We describe an efficienct chemical conversion platform with in situ immobilization of P450-BM3 on poly(3-hydroxybutyrate) granules. Through fusion with phasin, P450-BM3 is easily immobilized on poly(3-hydroxybutyrate) granules in Escherichia coli. In our work, the immobilized P450 exhibited higher stability and catalytic activity compared to free P450 against changes of pH, temperature, and concentrations of urea and ions. Through quick recovery of immobilized enzyme, the P450-P(3HB) complex successfully catalyzed an O-dealkylation reaction several times with maintained activity. Using the robust P450-P(3HB) complex, we performed a P450-catalyzed reaction on a preparative reactor scale (100 mL) and high-level production (12.3 ?M) of 7-hydroxycoumarine from 7-ethoxycoumarin could be achieved.
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Identification of recurrent FGFR3-TACC3 fusion oncogenes from lung adenocarcinoma.
Clin. Cancer Res.
PUBLISHED: 10-07-2014
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Purpose: Targetable oncogenic alterations are detected more commonly in patients with non-small cell lung cancer (NSCLC) who never smoked cigarettes. For such patients, specific kinase inhibitors have emerged as effective clinical treatments. However, the currently known oncogenic alterations do not account for all never smokers who develop NSCLC. We sought to identify additional oncogenic alterations from patients with NSCLC in order to define additional treatment options. Experimental Design: We analyzed 576 lung adenocarcinomas from patients of Asian and Caucasian ethnicity. We identified a subset of cancers that did not harbor any known oncogenic alteration. We performed targeted next generation sequencing (NGS) assay on 24 patients from this set with > 75% tumor cell content. Results: EGFR mutations were the most common oncogenic alteration from both Asian (53%) and Caucasian (41.6%) patients. No known oncogenic alterations were present in 25.7% of Asian and 31% of Caucasian tumor specimens. We identified a FGFR3-TACC3 fusion event in 1 of 24 patients from this subset using targeted NGS. Two additional patients harboring FGFR3-TACC3 were identified by screening our entire cohort (overall prevalence: 0.5%). Expression of FGFR3-TACC3 led to IL-3 independent growth in Ba/F3 cells. These cells were sensitive to pan-FGFR inhibitors but not the EGFR inhibitor gefitinib. Conclusions: FGFR3-TACC3 rearrangements occur in a subset of patients with lung adenocarcinoma. Such patients should be considered for clinical trials featuring FGFR inhibitors.
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Herniated intervertebral disk induces hypertrophy and ossification of ligamentum flavum.
J Spinal Disord Tech
PUBLISHED: 09-25-2014
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In vitro experiment using degenerated human ligamentum flavum (LF) and herniated intervertebral disk (IVD).
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[Mediating effect of self-efficacy in the relationship between anger and functional health of homeless men].
J Korean Acad Nurs
PUBLISHED: 09-19-2014
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The purpose of this study was to examine the mediation of self-efficacy in the relationship between anger and the functional health of homeless men in order to provide a basis for planning nursing interventions to improve the functional health of homeless persons.
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Prospective randomized trial comparing pushable coil and detachable coil during percutaneous implantation of port-catheter system for hepatic artery infusion chemotherapy.
Abdom Imaging
PUBLISHED: 09-13-2014
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The purpose of this study was to prospectively compare the efficacy and controllability of pushable coil and detachable coil during embolization of gastroduodenal artery (GDA) while performing percutaneous implantation of port-catheter system for hepatic artery infusion chemotherapy. Fifty patients (M:F = 42:8, age: 31-81 years) with advanced hepatocellular carcinoma undergoing port-catheter system implantation were randomized into pushable coil group and detachable coil group. During catheter fixation, GDA was embolized as close to the origin as possible. Success rate, number of coils used, number of coils removed due to malposition after deployment, time to occlusion, uncoiled GDA length, pushability, and complications were compared. Pushability was graded as no tension, slight tension, and difficult to advance. Embolization was successful in 49 patients. One failure resulted from repeated regurgitation of pushable coil into hepatic artery. Number of coils used and removed coils, time to occlusion, and uncoiled GDA length were 1-3 (mean 2.32), 5 coils in 3 patients, 4-20 min (mean 8.00), and 0-15.0 mm (mean 3.36) in pushable coil group, and 1-5 (mean 2.12), 2 coils in 2 patients, 2-15 min (mean 7.40), and 0-10.2 mm (mean 2.92) in detachable coil group, respectively, without significant difference. Pushability was no tension (n = 24) and slight tension (n = 1) in pushable coil group and no tension (n = 16), slight tension (n = 7), and difficult to advance (n = 2) in detachable coil group. One hepatic artery dissection occurred in the failed case during coil removal. Pushable coils and detachable coils had similar efficacy and controllability during GDA embolization, although there was a trend favoring detachable coil.
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High-level conversion of L-lysine into 5-aminovalerate that can be used for nylon 6,5 synthesis.
Biotechnol J
PUBLISHED: 09-08-2014
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L-Lysine is a potential feedstock for the production of bio-based precursors for engineering plastics. In this study, we developed a microbial process for high-level conversion of L-lysine into 5-aminovalerate (5AVA) that can be used as a monomer in nylon 6,5 synthesis. Recombinant Escherichia coli WL3110 strain expressing Pseudomonas putida delta-aminovaleramidase (DavA) and lysine 2-monooxygenase (DavB) was grown to high density in fed-batch culture and used as a whole cell catalyst. High-density E. coli WL3110 expressing DavAB, grown to an optical density at 600 nm (OD600 ) of 30, yielded 36.51 g/L 5AVA from 60 g/L L-lysine in 24 h. Doubling the cell density of E. coli WL3110 improved the conversion yield to 47.96 g/L 5AVA from 60 g/L of L-lysine in 24 h. 5AVA production was further improved by doubling the L-lysine concentration from 60 to 120 g/L. The highest 5AVA titer (90.59 g/L; molar yield 0.942) was obtained from 120 g/L L-lysine by E. coli WL3110 cells grown to OD600 of 60. Finally, nylon 6,5 was synthesized by bulk polymerization of ?-caprolactam and ?-valerolactam prepared from microbially synthesized 5AVA. The hybrid system demonstrated here has promising possibilities for application in the development of industrial bio-nylon production processes.
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Acupuncture for the treatment of spasticity after stroke: a meta-analysis of randomized controlled trials.
J Altern Complement Med
PUBLISHED: 09-06-2014
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Acupuncture has been suggested as a treatment for spasticity in patients with stroke. The available literature was reviewed in an effort to assess its efficacy in this situation.
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Over-expression of Roquin aggravates T cell mediated hepatitis in transgenic mice using T cell specific promoter.
Biochem. Biophys. Res. Commun.
PUBLISHED: 08-22-2014
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Chronic hepatitis is a major cause of liver cancer, so earlier treatment of hepatitis might be reducing liver cancer incidence. Hepatitis can be induced in mice by treatment with Concanavalin A (Con A); the resulting liver injury causes significant CD4(+) T cell activation and infiltration. In these T cells, Roquin, a ring-type E3 ubiquitin ligase, is activated. To investigate the role of Roquin, we examined Con A-induced liver injury and T cell infiltration in transgenic (Tg) mice overexpressing Roquin specifically in T cells. In Roquin Tg mice, Con A treatment caused greater increases in both the levels of liver injury enzymes and liver tissue apoptosis, as revealed by TUNEL and H&E staining, than wild type (WT) mice. Further, Roquin Tg mice respond to Con A treatment with greater increases in the T cell population, particularly Th17 cells, though Treg cell counts are lower. Roquin overexpression also enhances increases in pro-inflammatory cytokines, including IFN-?, TNF-? and IL-6, upon liver injury. Furthermore, Roquin regulates the immune response and apoptosis in Con A induced hepatitis via STATs, Bax and Bcl2. These findings suggest that over-expression of Roquin exacerbates T-cell mediated hepatitis.
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Gestational loss and growth restriction by angiogenic defects in placental growth factor transgenic mice.
Arterioscler. Thromb. Vasc. Biol.
PUBLISHED: 08-21-2014
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Angiogenesis is an important biological process during development, reproduction, and in immune responses. Placental growth factor (PlGF) is a member of vascular endothelial growth factor that is critical for angiogenesis and vasculogenesis. We generated transgenic mice overexpressing PlGF in specifically T cells using the human CD2-promoter to investigate the effects of PlGF overexpression.
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Plasminogen activator inhibitor-1 as an early potential diagnostic marker for Alzheimer's disease.
Exp. Gerontol.
PUBLISHED: 08-06-2014
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Alzheimer's disease (AD) is the most common cause of dementia in individuals over 65years old. However, to date, no useful early diagnostic markers for AD have been discovered. We examined the utility of plasminogen activator inhibitor-1 (PAI-1) as a potential biomarker for AD in subjects with mild cognitive impairment (MCI) or AD, as well as in nondemented healthy controls. Plasma PAI-1 levels were measured by enzyme-linked immunosorbent assays (ELISAs) in samples collected from 76 patients with MCI, 74 patients with AD, and 76 healthy controls. Our results show that plasma PAI-1 levels gradually increased as dementia progressed. The mean levels of plasma PAI-1 in patients with MCI and AD patients were significantly higher than those of in healthy controls. Consistently, neuropsychological examination (e.g., MMSE, CDR) also demonstrated significant correlations between the plasma PAI-1 levels and cognitive function. In conclusion, the level of plasma PAI-1 is a potential biomarker for the early detection and diagnosis of AD.
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Development of multiplex PCR assay for simultaneous detection of Salmonella genus, Salmonella subspecies I, S. Enteritidis, S. Heidelberg, and S. Typhimurium.
J. Appl. Microbiol.
PUBLISHED: 07-30-2014
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The aim of this research was to develop multiplex PCR assay that could simultaneously detect Salmonella genus, Salmonella subsp. I, S. Enteritidis, Heidelberg, and Typhimurium since these Salmonella serovars are the most common isolates associated with poultry products.
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Biocatalytic conversion of methane to methanol as a key step for development of methane-based biorefineries.
J. Microbiol. Biotechnol.
PUBLISHED: 07-25-2014
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Methane is considered as a next generation carbon feedstock due to vast reserves of natural and shale gas. Methane can be converted to methanol by various methods, which in turn can be used as a starting chemical for the production of value-added chemicals using existing chemical conversion processes. Methane monooxygenase is the key enzyme that catalyzes addition of oxygen to methane. Methanotrophic bacteria can transform methane to methanol by inhibiting methanol dehydrogenase. In this paper, we review the recent progress on biocatalytic conversion of methane to methanol as a key step for methane-based refinery systems and discuss future prospects for this technology.
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Associations between estimated desaturase activity and insulin resistance in korean boys.
Osong Public Health Res Perspect
PUBLISHED: 07-21-2014
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Obesity in childhood increases the risk of obesity in adulthood, and is predictive of the development of metabolic disorders. The fatty acid compositions of various tissues, including blood, are associated with obesity and obesity-associated disorders. Thus, tracking plasma phospholipid (PL) features and metabolic parameters in young individuals may strengthen the utility of fatty acid composition as an early biomarker of future metabolic disorders.
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Video-assisted thoracic surgery for bronchogenic cysts: is this the surgical approach of choice?
Interact Cardiovasc Thorac Surg
PUBLISHED: 07-19-2014
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Although there is no consensus on the management of bronchogenic cyst, most surgeons advocate early removal, even in asymptomatic patients. To evaluate the feasibility and safety of video-assisted thoracic surgery (VATS) in the management of bronchogenic cysts and long-term follow-up, a retrospective analysis was performed.
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Oncologic Outcomes According to Remnant Lymph Node Metastases in Pathologic T0 (ypT0) Esophageal Squamous Cell Carcinoma Following Prospective Neoadjuvant Therapy and Surgery.
Ann. Surg. Oncol.
PUBLISHED: 07-17-2014
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Some esophageal cancer patients may have residual nodal metastases following pathologic complete response (pCR) of the primary tumor after neoadjuvant chemoradiotherapy (CRT). This study examines the prognosis according to lymph node metastases in pathologic T0 patients who received neoadjuvant CRT followed by surgery for esophageal squamous cell carcinoma (SCC).
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Baicalein inhibits agonist- and tumor cell-induced platelet aggregation while suppressing pulmonary tumor metastasis via cAMP-mediated VASP phosphorylation along with impaired MAPKs and PI3K-Akt activation.
Biochem. Pharmacol.
PUBLISHED: 07-17-2014
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Recently, the importance of platelet activation in cancer metastasis has become generally accepted. As a result, the development of new platelet inhibitors with minimal adverse effects is now a promising area of targeted cancer therapy. Baicalein is a functional ingredient derived from the root of Scutellaria baicalensis Georgi, a plant used intraditional medicine. The pharmacological effects of this compound including anti-oxidative and anti-inflammatory activities have already been demonstrated. However, its effects on platelet activation are unknown. We therefore investigated the effects of baicalein on ligand-induced platelet aggregation and pulmonary cancer metastasis. In the present study, baicalein inhibited agonist-induced platelet aggregation, granule secretion markers (P-selectin expression and ATP release), [Ca(2+)]i mobilization, and integrin ?IIb?3 expression. Additionally, baicalein attenuated ERK2, p38, and Akt activation, and enhanced VASP phosphorylation. Indeed, baicalein was shown to directly inhibit PI3K kinase activity. Moreover, baicalein attenuated the platelet aggregation induced by C6 rat glioma tumor cells in vitro and suppressed CT26 colon cancer metastasis in mice. These features indicate that baicalein is a potential therapeutic drug for the prevention of cancer metastasis.
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Value of Flexible Bronchoscopy for the Preoperative Assessment of NSCLC Diagnosed Using Percutaneous Core Needle Biopsy.
Thorac Cardiovasc Surg
PUBLISHED: 07-15-2014
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Background?We aimed to investigate the value of routine flexible bronchoscopy (FB) for the preoperative assessment of early-stage non-small cell lung cancer (NSCLC) diagnosed using percutaneous core needle biopsy (PCNB). Methods?We enrolled 688 NSCLC patients who were treated at our hospital between January 2003 and December 2012 and who met the following criteria: (1) early-stage lung cancer (stage I or II); (2) lung cancer had been diagnosed using PCNB; and (3) no evidence of endobronchial disease in the airways other than the primary cancer site on both chest computed tomography (CT) and positron emission tomography-CT (PET-CT). All NSCLC patients were from the same tertiary referral center, where FB is routinely performed preoperatively for this disease, and their medical records were reviewed retrospectively. Results?Of the 688 patients included in the study, 451 (65.6%) were male and the median age was 65 years. Pathology analysis revealed that adenocarcinoma was the most frequently observed cell type (516/688, 75.0%). The distribution of preoperative clinical staging for the 688 patients was (1) IA (54.5%, 375/688); (2) IB (22.1%, 152/688); (3) IIA (18.2%, 125/688); and (4) IIB (5.2%, 36/688). The majority of these patients (95.2%, 655/688) underwent surgical resection. Unsuspected malignant endobronchial lesion on FB was found in only two cases (0.3%), and the surgical strategy had to be modified for both of these patients. Conclusion?Preoperative FB is not beneficial for screening the airways of almost any patient with early-stage NSCLC, provided that neither PET-CT nor CT reveal any evidence of endobronchial malignant involvement other than at the primary cancer site.
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Metabolic engineering of Ralstonia eutropha for the production of polyhydroxyalkanoates from sucrose.
Biotechnol. Bioeng.
PUBLISHED: 07-11-2014
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A sucrose utilization pathway was established in Ralstonia eutropha NCIMB11599 and R. eutropha 437-540 by introducing the Mannheimia succiniciproducens MBEL55E sacC gene that encodes ?-fructofuranosidase. These engineered strains were examined for the production of poly(3-hydroxybutyrate) [P(3HB)] and poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)], respectively, from sucrose as a carbon source. It was found that ?-fructofuranosidase excreted into the culture medium could hydrolyze sucrose to glucose and fructose, which were efficiently used as carbon sources by recombinant R. eutropha strains. When R. eutropha NCIMB11599 expressing the sacC gene was cultured in nitrogen-free chemically defined medium containing 20?g/L of sucrose, a high P(3HB) content of 73.2?wt% could be obtained. In addition, R. eutropha 437-540 expressing the Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene accumulated P(3HB-co-21.5?mol% LA) to a polymer content of 19.5?wt% from sucrose by the expression of the sacC gene and the Escherichia coli ldhA gene. The molecular weights of P(3HB) and P(3HB-co-21.5?mol%LA) synthesized in R. eutropha using sucrose as a carbon source were 3.52?×?10(5) (Mn ) and 2.19?×?10(4) (Mn ), respectively. The engineered R. eutropha strains reported here will be useful for the production of polyhydroxyalkanoates (PHAs) from sucrose, one of the most abundant and relatively inexpensive carbon sources. Biotechnol. Bioeng. 2014;9999: 1-6. © 2014 Wiley Periodicals, Inc.
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Crystallization and preliminary X-ray analysis of the C-terminal fragment of Ski7 from Saccharomyces cerevisiae.
Acta Crystallogr F Struct Biol Commun
PUBLISHED: 07-04-2014
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Ski7 (superkiller protein 7) plays a critical role in the mRNA surveillance pathway. The C-terminal fragment of Ski7 (residues 520-747) from Saccharomyces cerevisiae was heterologously expressed in Escherichia coli and purified to homogeneity. It was successfully crystallized and preliminary X-ray data were collected to 2.0?Å resolution using synchrotron radiation. The crystal belonged to a trigonal space group, either P3121 or P3221, with unit-cell parameters a = b = 73.5, c = 83.6?Å. The asymmetric unit contains one molecule of the C-terminal fragment of Ski7 with a corresponding crystal volume per protein mass (VM) of 2.61?Å(3)?Da(-1) and a solvent content of 52.8% by volume. The merging R factor is 6.6%. Structure determination by MAD phasing is under way.
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Relationship between Serum Levels of Body Iron Parameters and Insulin Resistance and Metabolic Syndrome in Korean Children.
Osong Public Health Res Perspect
PUBLISHED: 06-19-2014
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An increase in serum ferritin and levels of the cleaved soluble form of transferrin receptor (sTfR) are related to several metabolic conditions. We evaluated the relationship between body iron status indicators, including ferritin and sTfR, and insulin resistance and metabolic syndrome (MetS) in Korean children.
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Association between Dyslipidemia and the Prevalence of Colon Polyps Based on a Health Evaluation of Subjects at a Hospital.
Korean J Fam Med
PUBLISHED: 05-22-2014
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Colonic neoplasm is associated with western diet intake and physical inactivity. These life styles are also risk factors for dyslipidemia and metabolic syndrome. The aim of this study was to evaluate the association between dyslipidemia and the prevalence of colon polyps including colon adenoma as a precancerous lesion of colonic neoplasms.
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Effects of phosphoglucomutase gene (PGM) in Streptococcus parauberis on innate immune response and pathogenicity of olive flounder (Paralichthys olivaceus).
Fish Shellfish Immunol.
PUBLISHED: 05-20-2014
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In recent years, Streptococcus parauberis infection has been an emerging problem in aquaculture in South Korea because of its more frequent isolation than other streptococcal bacteria including Streptococcus iniae. To develop effective treatment and prophylaxis methods against this emerging disease by S. parauberis, it is necessary to understand the underlying pathogenic mechanisms. To uncover the pathogenicity, the mutant strain of S. parauberis with a deleted phosphoglucomutase (PGM) gene which has been known to be an important virulence factor in bacterial pathogens was generated to investigate the relationship between virulence and gene function using an allelic exchange mutagenesis method. Allelic exchange mutagenesis of the phosphoglucomutase gene resulted in phenotype changes including decreased extracellular capsules, reduced buoyancy, increased hydrophobicity and reduced growth. Moreover, the S. parauberis mutant was more sensitive to innate immune clearance mechanisms including serum, mucus and phagocyte killing and could not induce mortality in olive flounder. These phenotype changes and the attenuated virulence of the pathogen to fish could be due to the reduction in capsule production by mutation of the PGM gene. The results provide evidences that phosphoglucomutase expression contributes to S. parauberis virulence in fish by affecting bacterial survival against the host's humoral and cellular defense mechanisms.
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Purification, characterization and preliminary X-ray crystallographic studies of monodehydroascorbate reductase from Oryza sativa L. japonica.
Acta Crystallogr F Struct Biol Commun
PUBLISHED: 05-13-2014
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Monodehydroascorbate reductase (MDHAR; EC 1.6.5.4) is a key enzyme in the reactive oxygen species (ROS) detoxification system of plants. The participation of MDHAR in ascorbate (AsA) recycling in the ascorbate-glutathione cycle is important in the acquired tolerance of crop plants to abiotic environmental stresses. Thus, MDHAR represents a strategic target protein for the improvement of crop yields. Although physiological studies have intensively characterized MDHAR, a structure-based functional analysis is not available. Here, a cytosolic MDHAR (OsMDHAR) derived from Oryza sativa L. japonica was expressed using Escherichia coli strain NiCo21 (DE3) and purified. The purified OsMDHAR showed specific enzyme activity (approximately 380?U per milligram of protein) and was crystallized using the hanging-drop vapour-diffusion method at pH 8.0 and 298?K. The crystal diffracted to 1.9?Å resolution and contained one molecule in the asymmetric unit (the Matthews coefficient VM is 1.98?Å(3)?Da(-1), corresponding to a solvent content of 38.06%) in space group P41212 with unit-cell parameters a = b = 81.89, c = 120.4?Å. The phase of the OsMDHAR structure was resolved by the molecular-replacement method using a ferredoxin reductase from Acidovorax sp. strain KKS102 (PDB entry 4h4q) as a model.
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Dissecting the role of bone marrow stromal cells on bone metastases.
Biomed Res Int
PUBLISHED: 05-09-2014
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Tumor-induced bone disease is a dynamic process that involves interactions with many cell types. Once metastatic cancer cells reach the bone, they are in contact with many different cell types that are present in the cell-rich bone marrow. These cells include the immune cells, myeloid cells, fibroblasts, osteoblasts, osteoclasts, and mesenchymal stem cells. Each of these cell populations can influence the behavior or gene expression of both the tumor cells and the bone microenvironment. Additionally, the tumor itself can alter the behavior of these bone marrow cells which further alters both the microenvironment and the tumor cells. While many groups focus on studying these interactions, much remains unknown. A better understanding of the interactions between the tumor cells and the bone microenvironment will improve our knowledge on how tumors establish in bone and may lead to improvements in diagnosing and treating bone metastases. This review details our current knowledge on the interactions between tumor cells that reside in bone and their microenvironment.
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Sulforaphane induces autophagy through ERK activation in neuronal cells.
FEBS Lett.
PUBLISHED: 04-16-2014
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Sulforaphane (SFN), an activator of nuclear factor E2-related factor 2 (Nrf2), has been reported to induce autophagy in several cells. However, little is known about its signaling mechanism of autophagic induction. Here, we provide evidence that SFN induces autophagy with increased levels of LC3-II through extracellular signal-regulated kinase (ERK) activation in neuronal cells. Pretreatment with NAC (N-acetyl-l-cysteine), a well-known antioxidant, completely blocked the SFN-induced increase in LC3-II levels and activation of ERK. Knockdown or overexpression of Nrf2 did not affect autophagy. Together, the results suggest that SFN-mediated generation of reactive oxygen species (ROS) induces autophagy via ERK activation, independent of Nrf2 activity in neuronal cells.
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Opto-mechanical analysis of nonlinear elastomer membrane deformation under hydraulic pressure for variable-focus liquid-filled microlenses.
Opt Express
PUBLISHED: 03-26-2014
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Nonlinear large deformation of a transparent elastomer membrane under hydraulic pressure was analyzed to investigate its optical performance for a variable-focus liquid-filled membrane microlens. In most membrane microlenses, actuators control the hydraulic pressure of optical fluid so that the elastomer membrane together with the internal optical fluid changes its shape, which alters the light path of the microlens to adapt its optical power. A fluid-structure interaction simulation was performed to estimate the transient behavior of the microlens under the operation of electroactive polymer actuators, demonstrating that the viscosity of the optical fluid successfully stabilizes the fluctuations within a fairly short period of time during dynamic operations. Axisymmetric nonlinear plate theory was used to calculate the deformation profile of the membrane under hydrostatic pressure, with which optical characteristics of the membrane microlens were estimated. The effects of gravitation and viscoelastic behavior of the elastomer membrane on the optical performance of the membrane microlens were also evaluated with finite element analysis.
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Effect of novel cyclohexane diester and benzene diester derivatives on melanogenesis.
J Cosmet Sci
PUBLISHED: 03-20-2014
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In order to investigate potent whitening agents, we synthesized 15 cyclohexane diester derivatives and 15 benzene diester derivatives. To evaluate their structure-cytotoxicity relationships, we performed cell cytotoxicity tests on B16F10 mouse melanoma cells. To understand their whitening effects, melanin synthesis inhibitory activities in B16F10 cells and mushroom tyrosinase inhibitory activities were performed. In most cases, cell cytotoxicity was observed to be lower in 1,3-diester than in 1,2- and 1,4-diesters; when it came to the structural isomer of the side chain, all derivatives except the 1,2-cyclohexane diester derivatives showed lower cell cytotoxicity in the branch type of the side chain than in the linear type. Among the compounds evaluated, the compounds cyclohexane-1,3-diyl bis(decanoate), cyclohexane-1,4-diyl dioctanoate, and 1,3-phenylene bis (2-ethylhexanoate) emerged as potent melanin synthesis inhibitors. Our goal was to determine the expression levels of proteins involved in melanogenesis, Western blotting and RT-PCR showing that these compounds decreased tyrosinase, TRP-1, and TRP-2 while demonstrating significantly low cytotoxicity.
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Efficient extracellular production of type I secretion pathway-dependent Pseudomonas fluorescens lipase in recombinant Escherichia coli by heterologous ABC protein exporters.
Biotechnol. Lett.
PUBLISHED: 03-19-2014
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Heterologous ABC protein exporters, the apparatus of type I secretion pathway in Gram-negative bacteria, were used for extracellular production of Pseudomonas fluorescens lipase (TliA) in recombinant Escherichia coli. The effect of the expression of different ABC protein exporter gene clusters (P. fluorescens tliDEF, Pseudomonas aeruginosa aprDEF, Erwinia chrysanthemi prtDEF, and Serratia marcescens lipBCD genes) was examined on the secretion of TliA at growth temperatures of 20, 25, 30 and 35 °C. TliA secretion in recombinant E. coli XL10-Gold varied depending upon type of ABC protein exporter and culture temperature. E. coli expressing S. marcescens lipBCD genes showed the highest secretion level of TliA (122.8 U ml(-1)) when cultured at 25 °C. Thus, optimized culture conditions for efficient extracellular production of lipase in recombinant E. coli can be designed by changing the type of ABC protein exporter and the growth temperature.
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Correlations Between Electrically Quantified Pain Degree, Subjectively Assessed Visual Analogue Scale, and the McGill Pain Questionnaire: A Pilot Study.
Ann Rehabil Med
PUBLISHED: 03-12-2014
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To evaluate the clinical utility of the electrically calculated quantitative pain degree (QPD) and to correlate it with subjective assessments of pain degree including a visual analogue scale (VAS) and the McGill Pain Questionnaire (MPQ).
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Postoperative radiation therapy following the incomplete resection of a non-small cell lung cancer.
Radiat Oncol J
PUBLISHED: 03-05-2014
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To review the results of postoperative radiation therapy (PORT) for residual non-small cell lung cancer (NSCLC) following surgical resection and evaluate multiple clinicopathologic prognostic factors.
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Comparison of the Efficacy of Covered versus Uncovered Metallic Stents in Treating Inoperable Malignant Common Bile Duct Obstruction: A Randomized Trial.
J Vasc Interv Radiol
PUBLISHED: 03-04-2014
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To compare patency and overall survival achieved with covered versus uncovered metallic stents among patients with inoperable malignant extrahepatic biliary obstruction.
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Crystallization and preliminary X-ray crystallographic studies of dehydroascorbate reductase (DHAR) from Oryza sativa L. japonica.
Acta Crystallogr F Struct Biol Commun
PUBLISHED: 02-25-2014
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Dehydroascorbate reductase from Oryza sativa L. japonica (OsDHAR), a key enzyme in the regeneration of vitamin C, maintains reduced pools of ascorbic acid to detoxify reactive oxygen species. In previous studies, the overexpression of OsDHAR in transgenic rice increased grain yield and biomass as well as the amount of ascorbate, suggesting that ascorbate levels are directly associated with crop production in rice. Hence, it has been speculated that the increased level of antioxidants generated by OsDHAR protects rice from oxidative damage and increases the yield of rice grains. However, the crystal structure and detailed mechanisms of this important enzyme need to be further elucidated. In this study, recombinant OsDHAR protein was purified and crystallized using the sitting-drop vapour-diffusion method at pH 8.0 and 298?K. Plate-shaped crystals were obtained using 0.15?M potassium bromide, 30%(w/v) PEG MME 2000 as a precipitant, and the crystals diffracted to a resolution of 1.9?Å on beamline 5C at the Pohang Accelerator Laboratory. The X-ray diffraction data indicated that the crystal contained one OsDHAR molecule in the asymmetric unit and belonged to space group P2? with unit-cell parameters a=47.03, b=48.38, c=51.83?Å, ?=107.41°.
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Surgical management of pulmonary adenocarcinoma presenting as a pure ground-glass nodule.
Eur J Cardiothorac Surg
PUBLISHED: 02-23-2014
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With recent advances in radiology, the detection of ground-glass nodules (GGNs) has become increasingly common. However, there still is no consensus on management, especially on the need for systemic lymph node (LN) dissection. The purpose of this study was to evaluate the surgical outcomes on the basis of the extent of resection of the primary lesion and mediastinal LN dissection and to carefully suggest appropriate treatment strategies in the patients with pulmonary adenocarcinoma presenting as pure ground-glass opacities.
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Primary Ewing's Sarcoma of the Lung.
Korean J Thorac Cardiovasc Surg
PUBLISHED: 02-05-2014
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Most cases of Ewing's sarcoma are reported in the bone, and extraosseous Ewing's sarcoma is an extremely rare disease. Here, we report a rare case of primary pulmonary Ewing's sarcoma in a patient with hemoptysis. The patient underwent right upper lung lobe lobectomy with adjuvant chemotherapy and radiation therapy and has been free of recurrent disease for 4 years.
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Modern outcome and risk analysis of surgically resected occult N2 non-small cell lung cancer.
Ann. Thorac. Surg.
PUBLISHED: 01-14-2014
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This study was performed to assess the incidence, survival, and risk factors associated with unsuspected pathologic N2 disease in patients with resectable clinical N0-1 non-small cell lung cancer.
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Osteal macrophages support physiologic skeletal remodeling and anabolic actions of parathyroid hormone in bone.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 01-09-2014
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Cellular subpopulations in the bone marrow play distinct and unexplored functions in skeletal homeostasis. This study delineated a unique role of osteal macrophages in bone and parathyroid hormone (PTH)-dependent bone anabolism using murine models of targeted myeloid-lineage cell ablation. Depletion of c-fms(+) myeloid lineage cells [via administration of AP20187 in the macrophage Fas-induced apoptosis (MAFIA) mouse model] reduced cortical and trabecular bone mass and attenuated PTH-induced trabecular bone anabolism, supporting the positive function of macrophages in bone homeostasis. Interestingly, using a clodronate liposome model with targeted depletion of mature phagocytic macrophages an opposite effect was found with increased trabecular bone mass and increased PTH-induced anabolism. Apoptotic cells were more numerous in MAFIA versus clodronate-treated mice and flow cytometric analyses of myeloid lineage cells in the bone marrow showed that MAFIA mice had reduced CD68(+) cells, whereas clodronate liposome-treated mice had increased CD68(+) and CD163(+) cells. Clodronate liposomes increased efferocytosis (clearance of apoptotic cells) and gene expression associated with alternatively activated M2 macrophages as well as expression of genes associated with bone formation including Wnt3a, Wnt10b, and Tgfb1. Taken together, depletion of early lineage macrophages resulted in osteopenia with blunted effects of PTH anabolic actions, whereas depletion of differentiated macrophages promoted apoptotic cell clearance and transformed the bone marrow to an osteogenic environment with enhanced PTH anabolism. These data highlight a unique function for osteal macrophages in skeletal homeostasis.
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Korean Red Ginseng saponin fraction modulates radiation effects on lipopolysaccharide-stimulated nitric oxide production in RAW264.7 macrophage cells.
J Ginseng Res
PUBLISHED: 01-03-2014
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In previous work, we reported that Korean Red Ginseng saponin fraction (RGSF) showed anti-inflammatory activities in vitro and in vivo.
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Synthetic biology platform of CoryneBrick vectors for gene expression in Corynebacterium glutamicum and its application to xylose utilization.
Appl. Microbiol. Biotechnol.
PUBLISHED: 01-03-2014
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Currently, the majority of tools in synthetic biology have been designed and constructed for model organisms such as Escherichia coli and Saccharomyces cerevisiae. In order to broaden the spectrum of organisms accessible to such tools, we established a synthetic biological platform, called CoryneBrick, for gene expression in Corynebacterium glutamicum as a set of E. coli-C. glutamicum shuttle vectors whose elements are interchangeable with BglBrick standard parts. C. glutamicum is an established industrial microorganism for the production of amino acids, proteins, and commercially promising chemicals. Using the CoryneBrick vectors, we showed various time-dependent expression profiles of a red fluorescent protein. This CoryneBrick platform was also applicable for two-plasmid expression systems with a conventional C. glutamicum expression vector. In order to demonstrate the practical application of the CoryneBrick vectors, we successfully reconstructed the xylose utilization pathway in the xylose-negative C. glutamicum wild type by fast BglBrick cloning methods using multiple genes encoding for xylose isomerase and xylulose kinase, resulting in a growth rate of 0.11?±?0.004 h(-1) and a xylose uptake rate of 3.35 mmol/gDW/h when 1 % xylose was used as sole carbon source. Thus, CoryneBrick vectors were shown to be useful engineering tools in order to exploit Corynebacterium as a synthetic platform for the production of chemicals by controllable expression of the genes of interest.
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Overexpression of Jazf1 reduces body weight gain and regulates lipid metabolism in high fat diet.
Biochem. Biophys. Res. Commun.
PUBLISHED: 12-16-2013
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Jazf1 is a 27kDa nuclear protein containing three putative zinc finger motifs that is associated with diabetes mellitus and prostate cancer; however, little is known about the role that this gene plays in regulation of metabolism. Recent evidence indicates that Jazf1 transcription factors bind to the nuclear orphan receptor TR4. This receptor regulates PEPCK, the key enzyme involved in gluconeogenesis. To elucidate Jazf1s role in metabolism, we fed a 60% fat diet for up to 15weeks. In Jazf1 overexpression mice, weight gain was found to be significantly decreased. The expression of Jazf1 in the liver also suppressed lipid accumulation and decreased droplet size. These results suggest that Jazf1 plays a critical role in the regulation of lipid homeostasis. Finally, Jazf1 may provide a new therapeutic target in the management of obesity and diabetes.
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[6]-Shogaol inhibits growth and induces apoptosis of non-small cell lung cancer cells by directly regulating Akt1/2.
Carcinogenesis
PUBLISHED: 11-26-2013
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Non-small cell lung cancer (NSCLC) is the leading cause of cancer mortality worldwide. Despite progress in developing chemotherapeutics for the treatment of NSCLC, primary and secondary resistance limits therapeutic success. NSCLC cells exhibit multiple mutations in the epidermal growth factor receptor (EGFR), which cause aberrant activation of diverse cell signaling pathways. Therefore, suppression of the inappropriate amplification of EGFR downstream signaling cascades is considered to be a rational therapeutic and preventive strategy for the management of NSCLC. Our initial molecular target-oriented virtual screening revealed that the ginger components, including [6]-shogaol, [6]-paradol and [6]-gingerol, seem to be potential candidates for the prevention and treatment of NSCLC. Among the compounds, [6]-shogaol showed the greatest inhibitory effects on the NSCLC cell proliferation and anchorage-independent growth. [6]-Shogaol induced cell cycle arrest (G1 or G2/M) and apoptosis. Furthermore, [6]-shogaol inhibited Akt kinase activity, a downstream mediator of EGFR signaling, by binding with an allosteric site of Akt. In NCI-H1650 lung cancer cells, [6]-shogaol reduced the constitutive phosphorylation of signal transducer and activator of transcription-3 (STAT3) and decreased the expression of cyclin D1/3, which are target proteins in the Akt signaling pathway. The induction of apoptosis in NCI-H1650 cells by [6]-shogaol corresponded with the cleavage of caspase-3 and caspase-7. Moreover, intraperitoneal administration of [6]-shogaol inhibited the growth of NCI-H1650 cells as tumor xenografts in nude mice. [6]-Shogaol suppressed the expression of Ki-67, cyclin D1 and phosphorylated Akt and STAT3 and increased terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positivity in xenograft tumors. The current study clearly indicates that [6]-shogaol can be exploited for the prevention and/or treatment of NSCLC.
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Gastric conduit cancer after oesophagectomy for oesophageal cancer: incidence and clinical implications.
Eur J Cardiothorac Surg
PUBLISHED: 10-24-2013
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Gastric conduit cancer (GCC), which is a carcinoma that arises in the gastric conduit after oesophagectomy, often negatively affects long-term survivors of oesophageal cancer. The aim of this study was to evaluate the incidence and clinical implications of GCC.
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Parathyroid hormone-related protein drives a CD11b+Gr1+ cell-mediated positive feedback loop to support prostate cancer growth.
Cancer Res.
PUBLISHED: 09-26-2013
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In the tumor microenvironment, CD11b(+)Gr1(+) bone marrow-derived cells are a predominant source of protumorigenic factors such as matrix metalloproteinases (MMP), but how distal tumors regulate these cells in the bone marrow is unclear. Here we addressed the hypothesis that the parathyroid hormone-related protein (PTHrP) potentiates CD11b(+)Gr1(+) cells in the bone marrow of prostate tumor hosts. In two xenograft models of prostate cancer, levels of tumor-derived PTHrP correlated with CD11b(+)Gr1(+) cell recruitment and microvessel density in the tumor tissue, with evidence for mediation of CD11b(+)Gr1(+) cell-derived MMP-9 but not tumor-derived VEGF-A. CD11b(+)Gr1(+) cells isolated from mice with PTHrP-overexpressing tumors exhibited relatively increased proangiogenic potential, suggesting that prostate tumor-derived PTHrP potentiates this activity of CD11b(+)Gr1(+) cells. Administration of neutralizing PTHrP monoclonal antibody reduced CD11b(+)Gr1(+) cells and MMP-9 in the tumors. Mechanistic investigations in vivo revealed that PTHrP elevated Y418 phosphorylation levels in Src family kinases in CD11b(+)Gr1(+) cells via osteoblast-derived interleukin-6 and VEGF-A, thereby upregulating MMP-9. Taken together, our results showed that prostate cancer-derived PTHrP acts in the bone marrow to potentiate CD11b(+)Gr1(+) cells, which are recruited to tumor tissue where they contribute to tumor angiogenesis and growth.
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Single-center experience in the endovascular management of isolated iliac artery aneurysm.
Acta Radiol
PUBLISHED: 08-23-2013
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Isolated iliac artery aneurysms (IAA) are relatively uncommon and represent 2-7% of all intra-abdominal aneurysms. Surgery is the gold standard treatment for IAA. However, endovascular stent-graft placement is gaining acceptance as an alternative to surgery especially in patients with high surgical risk.
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Right Adrenal Venography Findings correlated with C-arm CT for Selection During C-arm CT-assisted Adrenal Vein Sampling in Primary Aldosteronism.
Cardiovasc Intervent Radiol
PUBLISHED: 08-11-2013
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This study was designed to evaluate retrospectively the efficacy of C-arm CT to confirm right adrenal vein catheterization during adrenal vein sampling (AVS) and to correlate adrenal venography findings with C-arm CT and/or biochemical results for right adrenal vein selection.
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Metabolic engineering of Escherichia coli for biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from glucose.
Appl. Microbiol. Biotechnol.
PUBLISHED: 08-02-2013
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The Escherichia coli XL1-blue strain was metabolically engineered to synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] through 2-ketobutyrate, which is generated via citramalate pathway, as a precursor for propionyl-CoA. Two different metabolic pathways were examined for the synthesis of propionyl-CoA from 2-ketobutyrate. The first pathway is composed of the Dickeya dadantii 3937 2-ketobutyrate oxidase or the E. coli pyruvate oxidase mutant (PoxB L253F V380A) for the conversion of 2-ketobutyrate into propionate and the Ralstonia eutropha propionyl-CoA synthetase (PrpE) or the E. coli acetyl-CoA:acetoacetyl-CoA transferase for further conversion of propionate into propionyl-CoA. The second pathway employs pyruvate formate lyase encoded by the E. coli tdcE gene or the Clostridium difficile pflB gene for the direct conversion of 2-ketobutyrate into propionyl-CoA. As the direct conversion of 2-ketobutyrate into propionyl-CoA could not support the efficient production of P(3HB-co-3HV) from glucose, the first metabolic pathway was further examined. When the recombinant E. coli XL1-blue strain equipped with citramalate pathway expressing the E. coli poxB L253F V380A gene and R. eutropha prpE gene together with the R. eutropha PHA biosynthesis genes was cultured in a chemically defined medium containing 20 g/L of glucose as a sole carbon source, P(3HB-co-2.3 mol% 3HV) was produced up to the polymer content of 61.7 wt.%. Moreover, the 3HV monomer fraction in P(3HB-co-3HV) could be increased up to 5.5 mol% by additional deletion of the prpC and scpC genes, which are responsible for the metabolism of propionyl-CoA in host strains.
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Engineered fumarate sensing Escherichia coli based on novel chimeric two-component system.
J. Biotechnol.
PUBLISHED: 08-02-2013
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DcuS/DcuR two component system (TCS) was firstly employed for the expression of the gfp gene under the dcuB gene promoter in aerobic condition to develop high throughput screening system able to screen microorganisms producing high amount of fumarate. However, the DcuS/DcuR TCS could not produce a signal strong enough to mediate the expression of the gfp gene responding fumarate concentration. Thus, DcuS/DucR TCS was engineered by recruiting the EnvZ/OmpR system, the most-studied TCS in E. coli. A chimeric DcuS/EnvZ (DcuSZ) TCS was constructed by fusing the sensor histidine kinase of DcuS with the cytoplasmic catalytic domain of EnvZ, in which the expression of the gfp gene or the ompC gene was mediated by the ompC gene promoter through the cognate response regulator, OmpR. The output signals produced by the chimeric DcuSZ TCS were enough to detect fumarate concentration quantatively, in which the expressions of the gfp gene and the ompC gene were proportional to the fumarate concentration in the medium. Moreover, principal component analysis of C4-dicarboxylates showed that DcuSZ chimera was highly specific to fumarate but could also respond to other C4-dicarboxylates, which strongly suggests that TCS-based high throughput screening system able to screen microorganisms producing target chemicals can be developed.
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Esophageal Cancer Associated with Multiple Primary Cancers: Surgical Approaches and Long-term Survival.
Ann. Surg. Oncol.
PUBLISHED: 08-01-2013
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The presence of multiple primary cancers (MPCs) in patients with esophageal cancer often presents physicians with a difficult therapeutic decision, because little is known about the appropriate treatment and long-term survival. The purpose of this study was to evaluate appropriate surgical approaches and long-term survival after surgery for esophageal cancer associated with MPCs.
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Outcomes of endovascular treatment for aortic pseudoaneurysm in Behcets disease.
J. Vasc. Surg.
PUBLISHED: 07-15-2013
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To evaluate the effectiveness of endovascular stent grafting for surgical management of aortic pseudoaneurysm in patients with Behcets disease (BD).
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S1 pocket of glutamate carboxypeptidase II: a new binding site for amyloid-? degradation.
Biochem. Biophys. Res. Commun.
PUBLISHED: 07-12-2013
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We recently reported that glutamate carboxypeptidase II (GCPII) has a new physiological function degrading amyloid-? (A?), distinct from its own hydrolysis activity in N-acetyl-L-aspartyl-L-glutamate (NAAG); however, its underlying mechanism remains undiscovered. Using site-directed mutagenesis and S1 pocket-specific chemical inhibitor (compound 2), which was developed for the present study based on in sillico computational modeling, we discovered that the A? degradation occurs through S1 pocket but not through S1 pocket responsible for NAAG hydrolysis. Treatment with compound 2 prevented GCPII from A? degradation without any impairment in NAAG hydrolysis. Likewise, 2-PMPA (specific GCPII inhibitor developed targeting S1 pocket) completely blocked the NAAG hydrolysis without any effect on A? degradation. Pre-incubation with NAAG and A? did not affect A? degradation and NAAG hydrolysis, respectively. These data suggest that GCPII has two distinctive binding sites for two different substrates and that A? degradation occurs through binding to S1 pocket of GCPII.
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Chylothorax Complicating Pulmonary Resection for Lung Cancer: Effective Management and Pleurodesis.
Ann. Thorac. Surg.
PUBLISHED: 07-07-2013
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Chylothorax associated with pulmonary resection for lung cancer, although rare, must be considered as a potential complication during thoracic surgery. In the present study, we investigated the effectiveness of a conservative approach (diet or pleurodesis) to the management of chylothorax.
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Sociodemographic characteristics of underweight korean adults: Korea national health and nutrition examination survey, 2007-2010.
Korean J Fam Med
PUBLISHED: 06-27-2013
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Underweight is associated with increased risk of mortality and morbidity. It is reported that the prevalence of underweight is increasing among Korean young women. However, there have been few studies on sociodemographic factors related to being underweight. This study was conducted to elucidate the sociodemographic characteristics of Korean underweight adults.
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Ultrasound-guided subclavian catheterization in pediatric patients with a linear probe: a case series.
Korean J Anesthesiol
PUBLISHED: 06-24-2013
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Central venous catheterization (CVC) can be difficult, especially with pediatric patients in critical care. Accessing the subclavian vein (SCV) can cause serious complications, including pneumothorax, arterial puncture, and hemothorax. Recently, the ultrasonographic (USG) technique has gained popularity, but its efficiency is not yet confirmed. Subclavian venous catheterization (SCVC) through the supraclavicular approach (SCA) with USG or accessing the brachiocephalic vein through the infraclavicular approach (ICA) has been reported in the past. A useful technique is reported that involves the use of a 40 mm probe rather than the usual 25 mm probe in order to confirm the location of the needle while successfully performing subclavian venous catheterization in pediatric patients weighing 1.1 kg to 15.0 kg.
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Outcomes of endovascular management for complicated chronic type B aortic dissection: effect of the extent of stent graft coverage and anatomic properties of aortic dissection.
J Vasc Interv Radiol
PUBLISHED: 06-04-2013
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To assess the effect of the extent of stent graft coverage and anatomic properties of aortic dissection on the outcomes of thoracic endovascular aortic repair (TEVAR) for complicated chronic type B aortic dissection (CCBAD) in terms of survival, reintervention, and false lumen thrombosis.
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Metabolic engineering of Ralstonia eutropha for the biosynthesis of 2-hydroxyacid-containing polyhydroxyalkanoates.
Metab. Eng.
PUBLISHED: 05-12-2013
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Polyhydroxyalkanoates (PHAs) are bio-based and biodegradable polyesters synthesized by numerous microorganisms. PHAs containing 2-hydroxyacids as monomer units have attracted much attention, but their production has not been efficient. Here, we metabolically engineered Ralstonia eutropha strains for the in vivo synthesis of PHAs containing 2-hydroxyacids as monomers. This was accomplished by replacing the R. eutropha phaC gene in the chromosome with either the R. eutropha phaC S506G A510K gene, which contains two point mutations, or the Pseudomonas sp. MBEL 6-19 phaC1437 gene. In addition, the R. eutropha phaAB genes in the chromosome were replaced with the Clostridium propionicum pct540 gene. All of the engineered R. eutropha strains produced PHAs containing 2-hydroxyacid monomers, including lactate and 2-hydroxybutyrate (2HB), along with 3-hydroxybutyrate (3HB) and/or 3-hydroxyvalerate (3HV), when they were cultured in nitrogen-free medium containing 5g/L lactate or 4g/L 2HB and 20g/L glucose as carbon sources. Expression of the Escherichia coli ldhA gene in engineered R. eutropha strains allowed production of poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)] from glucose as the sole carbon source. This is the first report on the production of 2-hydroxyacid-containing PHAs by metabolically engineered R. eutropha.
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Causes of hand tingling in visual display terminal workers.
Ann Rehabil Med
PUBLISHED: 04-30-2013
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To offer the basic data about the causes and distribution of hand tingling, symptoms and physical findings, and pressure pain threshold in desk workers.
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Surgical Outcomes of Pulmonary Mucoepidermoid Carcinoma: A Review of 23 Cases.
Thorac Cardiovasc Surg
PUBLISHED: 04-25-2013
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Background The aim of the present study was to evaluate the oncologic characteristics of pulmonary mucoepidermoid carcinoma (PMEC) and the efficacy of surgical resections.Materials and Methods The surgery for PMEC was performed in 23 patients at Asan Medical Center from January 2000 to December 2010. They accounted for 0.8% of all surgically resected pulmonary neoplasm in the center. The medical records of these patients were reviewed retrospectively.Results This study group was composed of 13 males (56.5%) and 10 females (43.5%). Median age was 41 years (range, 10 to 75 years). Complete resection with systematic mediastinal lymph node dissection was performed in all patients. There were three postoperative complications: atelectasis in one patient and chylothorax in two patients. There was no postoperative mortality. The median follow-up duration was 68 months (range, 13 to 115 months). In one patient, recurrent disease was found 73 months after surgery. All patients were followed without mortality until the end of this study. The 5- and 8-year disease-free survival rates were 100 and 90.9%, respectively.Conclusion Complete surgical resection with systematic lymph node dissection is an effective treatment for patients with PMEC and provides favorable prognosis.
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A Unique Small Molecule Inhibitor of Enolase Clarifies Its Role in Fundamental Biological Processes.
ACS Chem. Biol.
PUBLISHED: 04-22-2013
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Enolase is a component of the glycolysis pathway and a "moonlighting" protein, with important roles in diverse cellular processes that are not related to its function in glycolysis. However, small molecule tools to probe enolase function have been restricted to crystallography or enzymology. In this study, we report the discovery of the small molecule "ENOblock", which is the first, nonsubstrate analogue that directly binds to enolase and inhibits its activity. ENOblock was isolated by small molecule screening in a cancer cell assay to detect cytotoxic agents that function in hypoxic conditions, which has previously been shown to induce drug resistance. Further analysis revealed that ENOblock can inhibit cancer cell metastasis in vivo. Moreover, an unexpected role for enolase in glucose homeostasis was revealed by in vivo analysis. Thus, ENOblock is the first reported enolase inhibitor that is suitable for biological assays. This new chemical tool may also be suitable for further study as a cancer and diabetes drug candidate.
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Metabolic engineering of Escherichia coli for biosynthesis of D-galactonate.
Bioprocess Biosyst Eng
PUBLISHED: 04-03-2013
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D-galactose is an attractive substrate for bioconversion. Herein, Escherichia coli was metabolically engineered to convert D-galactose into D-galactonate, a valuable compound in the polymer and cosmetic industries. D-galactonate productions by engineered E. coli strains were observed in shake flask cultivations containing 2 g L(-1) D-galactose. Engineered E. coli expressing gld coding for galactose dehydrogenase from Pseudomonas syringae was able to produce 0.17 g L(-1) D-galactonate. Inherent metabolic pathways for assimilating both D-galactose and D-galactonate were blocked to enhance the production of D-galactonate. This approach finally led to a 7.3-fold increase with D-galactonate concentration of 1.24 g L(-1) and yield of 62.0 %. Batch fermentation in 20 g L(-1) D-galactose of E. coli ?galK?dgoK mutant expressing the gld resulted in 17.6 g L(-1) of D-galactonate accumulation and highest yield of 88.1 %. Metabolic engineering strategy developed in this study could be useful for industrial production of D-galactonate.
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Oncogenic and drug-sensitive NTRK1 rearrangements in lung cancer.
Nat. Med.
PUBLISHED: 03-28-2013
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We identified new gene fusions in patients with lung cancer harboring the kinase domain of the NTRK1 gene that encodes the high-affinity nerve growth factor receptor (TRKA protein). Both the MPRIP-NTRK1 and CD74-NTRK1 fusions lead to constitutive TRKA kinase activity and are oncogenic. Treatment of cells expressing NTRK1 fusions with inhibitors of TRKA kinase activity inhibited autophosphorylation of TRKA and cell growth. Tumor samples from 3 of 91 patients with lung cancer (3.3%) without known oncogenic alterations assayed by next-generation sequencing or fluorescence in situ hybridization demonstrated evidence of NTRK1 gene fusions.
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Quantified high-throughput screening of Escherichia coli producing poly(3-hydroxybutyrate) based on FACS.
Appl. Biochem. Biotechnol.
PUBLISHED: 03-25-2013
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Here, we report on a highly sensitive method for the detection of P(3HB) accumulation in Escherichia coli cells based on the automated flow cytometry system using fluorescent dyes. E. coli containing P(3HB) were stained with either BODIPY or Nile red fluorescent dye, and their staining properties were analyzed under a variety of conditions. Compared with Nile red, BODIPY was much more sensitive in staining P(3HB) and overall demonstrated a more rapid staining of cells, a greater resistance to photobleaching, and greater cell viability. In addition, we also successfully monitored heterogeneity in P(3HB) accumulation within a cell population using BODIPY staining and flow cytometry. We believe this optimized staining method using BODIPY in combination with screening by high-speed flow cytometer will be helpful in the engineering of host cells toward an enhanced production of bioplastics.
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TRAIL regulates collagen production through HSF1-dependent Hsp47 expression in activated hepatic stellate cells.
Cell. Signal.
PUBLISHED: 03-22-2013
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Hsp47 is a collagen-specific molecular chaperone, whose activity has been implicated in liver fibrosis. In this study, we showed that TRAIL treatment inhibited Hsp47 expression in dose- and time-dependent manners, subsequently leading to the decrease of collagen production in activated human hepatic stellate LX-2 cells. Overexpression of Hsp47 in LX-2 cells acquired resistance for TRAIL-induced collagen reduction and conversely, siRNA suppression of Hsp47 enhanced the decrease of collagen production due to TRAIL treatment. Moreover, we found that Hsp47 expression was under the transcriptional control of heat shock factor (HSF) 1 which is highly located on nucleus in activated human hepatic stellate LX-2 cells. Treatment of LX-2 cells with TRAIL decreased the active trimer formation of HSF1, increased the dephosphorylation of HSF1 (Ser(230)), and enhanced the translocation of HSF1 into cytosol. The accumulated HSF1 in cytosol led to downregulation of Hsp47 expression, resulting in the reduction of collagen production. Consistently, HSF1 silencing by siRNA prevented Hsp47 induction and subsequent collagen production, whereas overexpression of HSF1 restored the expression level of Hsp47 as well as collagen production in response to TRAIL treatment in LX-2 cells. Taken together, our data suggested that TRAIL induced HSF1 inactivation, consequently leading to the suppression of Hsp47-dependent collagen production in activated human hepatic stellate cells. Therefore, this study suggests that TRAIL may be an effective strategy for antifibrotic therapy in liver fibrosis.
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Microbial production of lactate-containing polyesters.
Microb Biotechnol
PUBLISHED: 03-11-2013
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Due to our increasing concerns on environmental problems and limited fossil resources, biobased production of chemicals and materials through biorefinery has been attracting much attention. Optimization of the metabolic performance of microorganisms, the key biocatalysts for the efficient production of the desired target bioproducts, has been achieved by metabolic engineering. Metabolic engineering allowed more efficient production of polyhydroxyalkanoates, a family of microbial polyesters. More recently, non-natural polyesters containing lactate as a monomer have also been produced by one-step fermentation of engineered bacteria. Systems metabolic engineering integrating traditional metabolic engineering with systems biology, synthetic biology, protein/enzyme engineering through directed evolution and structural design, and evolutionary engineering, enabled microorganisms to efficiently produce natural and non-natural products. Here, we review the strategies for the metabolic engineering of microorganisms for the in vivo biosynthesis of lactate-containing polyesters and for the optimization of whole cell metabolism to efficiently produce lactate-containing polyesters. Also, major problems to be solved to further enhance the production of lactate-containing polyesters are discussed.
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Efficient gamma-aminobutyric acid bioconversion by employing synthetic complex between glutamate decarboxylase and glutamate/GABA antiporter in engineered Escherichia coli.
J. Ind. Microbiol. Biotechnol.
PUBLISHED: 03-07-2013
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Gamma-aminobutyric acid (GABA) is a precursor of one of the most promising heat-resistant biopolymers, Nylon-4, and can be produced by the decarboxylation of monosodium glutamate (MSG). In this study, a synthetic protein complex was applied to improve the GABA conversion in engineered Escherichia coli. Complexes were constructed by assembling a single protein-protein interaction domain SH3 to the glutamate decarboxylase (GadA and GadB) and attaching a cognate peptide ligand to the glutamate/GABA antiporter (GadC) at the N-terminus, C-terminus, and the 233rd amino acid residue. When GadA and GadC were co-overexpressed via the C-terminus complex, a GABA concentration of 5.65 g/l was obtained from 10 g/l MSG, which corresponds to a GABA yield of 93 %. A significant increase of the GABA productivity was also observed where the GABA productivity increased 2.5-fold in the early culture period due to the introduction of the synthetic protein complex. The GABA pathway efficiency and GABA productivity were enhanced by the introduction of the complex between Gad and glutamate/GABA antiporter.
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Activation of NF-kappa B signaling promotes growth of prostate cancer cells in bone.
PLoS ONE
PUBLISHED: 03-05-2013
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Patients with advanced prostate cancer almost invariably develop osseous metastasis. Although many studies indicate that the activation of NF-?B signaling appears to be correlated with advanced cancer and promotes tumor metastasis by influencing tumor cell migration and angiogenesis, the influence of altered NF-?B signaling in prostate cancer cells within boney metastatic lesions is not clearly understood. While C4-2B and PC3 prostate cancer cells grow well in the bone, LNCaP cells are difficult to grow in murine bone following intraskeletal injection. Our studies show that when compared to LNCaP, NF-?B activity is significantly higher in C4-2B and PC3, and that the activation of NF-?B signaling in prostate cancer cells resulted in the increased expression of the osteoclast inducing genes PTHrP and RANKL. Further, conditioned medium derived from NF-?B activated LNCaP cells induce osteoclast differentiation. In addition, inactivation of NF-?B signaling in prostate cancer cells inhibited tumor formation in the bone, both in the osteolytic PC3 and osteoblastic/osteoclastic mixed C4-2B cells; while the activation of NF-?B signaling in LNCaP cells promoted tumor establishment and proliferation in the bone. The activation of NF-?B in LNCaP cells resulted in the formation of an osteoblastic/osteoclastic mixed tumor with increased osteoclasts surrounding the new formed bone, similar to metastases commonly seen in patients with prostate cancer. These results indicate that osteoclastic reaction is required even in the osteoblastic cancer cells and the activation of NF-?B signaling in prostate cancer cells increases osteoclastogenesis by up-regulating osteoclastogenic genes, thereby contributing to bone metastatic formation.
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Expression of a lipase on the cell-surface of Escherichia coli using the OmpW anchoring motif and its application to enantioselective reactions.
Biotechnol. Lett.
PUBLISHED: 02-27-2013
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Microbial-surface display is the expression of proteins or peptides on the surface of cells by fusing an appropriate protein as an anchoring motif. Here, the outer membrane protein W (OmpW) was selected as a fusion partner for functional expression of Pseudomonas fluorescence SIK W1 lipase (TliA) on the cell-surface of Escherichia coli. Localization of the truncated OmpW-TliA fusion protein on the cell-surface was confirmed by immunoblotting and functional assay of lipase activity. Enantioselective hydrolysis of rac-phenylethyl butanoate by the displayed lipase resulted in optically active (R)-phenyl ethanol with 96% enantiomeric excess and 44% of conversion in 5 days. Thus, a small outer membrane protein OmpW, is a useful anchoring motif for displaying an active enzyme of ~50 kDa on the cell-surface and the surface-displayed lipase can be employed as an enantioselective biocatalyst in organic synthesis.
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