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Find video protocols related to scientific articles indexed in Pubmed.
miR-410 Inhibition Induces RPE Differentiation of Amniotic Epithelial Stem Cells via Overexpression of OTX2 and RPE65.
Stem Cell Rev
PUBLISHED: 10-30-2014
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The retinal pigment epithelium (RPE) is a highly specialized cell type located between the choroid and neural retina of the eye. RPE degeneration causes irreversible visual impairment, extending to blindness. Cell therapy has recently emerged as a potential therapeutic approach for retinal degeneration. MicroRNA-based differentiation of stem cells is a new strategy for producing tissue-specific cell types. In this study, we developed a novel microRNA-based strategy for RPE induction from human amniotic epithelial stem cells (AESCs). We identified microRNAs involved in RPE development in AESCs. Of 29 putative human RPE-relevant microRNAs, microRNA-410 (miR-410) was predicted to target multiple RPE development-relevant genes. Inhibition of miR-410 induces overexpression of immature and mature RPE-specific factors, including OTX2, RPE65, Bestrophin and EMMPRIN. These RPE-like cells were morphologically altered toward a cobblestone-like shape and were able to phagocytize microbeads. We showed that miR-410 directly regulates predicted target genes OTX2 and RPE65. Our microRNA-based strategy demonstrated RPE differentiation in AESCs by treatment of an antisense microRNA-410 (anti-miR-410), without the use of additional factors or exogenous transduction. These findings suggest that miR-410 inhibition can be a useful tool for directed cell differentiation and an attractive method for cell therapy in human retinal degenerative diseases.
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Immunolocalisation and oestrogen regulation of small proline-rich protein 2a protein in the mouse uterus.
Reprod. Fertil. Dev.
PUBLISHED: 08-29-2014
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Small proline-rich protein 2a (Sprr2a) is one of the structural components of the cornified keratinocyte cell envelope that contributes to form a protective barrier in the skin against dehydration and environmental stress. Interestingly, Sprr2a mRNA is detected in the mouse uterus and is regulated by 17?-oestradiol (E2). In the present study, we investigated the effects of E2 and oestrogenic compounds on the regulation and localisation of Sprr2a protein in the mouse uterus. Immunohistochemical staining revealed that Sprr2a protein is detected only in the adult uterus, and not in the ovary, oviduct or testis. We also demonstrated that Sprr2a protein is tightly regulated by E2 in the mouse uterus and exclusively detected in luminal and glandular epithelial cells. Furthermore, Sprr2a is dose-dependently induced by oestrogenic compounds such as bisphenol A and 4-tert-octylphenol. Collectively, our studies suggest that Sprr2a protein may have a unique function in physiological events in the mouse uterus and can be used as an indicator to detect compounds with oestrogenic activity in the mouse uterus.
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Generation of minicells from an endotoxin-free gram positive strain Corynebacterium glutamicum.
J. Microbiol. Biotechnol.
PUBLISHED: 08-18-2014
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Drug delivery systems (DDSs) incorporating bacterial minicells have been evaluated as a very powerful tool in view of biocompatibility. However, limited studies have been carried out on these systems, mainly using minicells from Salmonella sp. and Escherichia coli. Thus, we generated a new minicell-producing strain from an endotoxin-free Corynebacterium glutamicum by the inactivation of genes related to cell division. The two knock-out strains, ?parA and ?ncgl1366, showed distinct abilities to produce minicells. The resulting minicells were purified via sequential antibiotic treatments and centrifugations, which resulted in reproducible yield.
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Improved pretreatment process using an electron beam for optimization of glucose yield with high selectivity.
Appl. Biochem. Biotechnol.
PUBLISHED: 08-16-2014
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In this study, electron beam irradiation (EBI) assisted by a dilute acid pretreatment process was investigated to improve the glucose yield and show high selectivity in the enzymatic hydrolysis of rice straw. In the first step, EBI of rice straw was performed at various doses ranging from 50 to 500 kGy. The electron beam-irradiated rice straw was then autoclaved with 3 % dilute acid at 120 °C for 1 h. The pretreated rice straw was finally subjected to enzymatic hydrolysis at 50 °C for 24, 48, and 72 h by 70 filter paper units (FPU)/mL cellulase and 40 cellobiose units (CbU)/mL glucosidase. Glucose was obtained with a very high selectivity of 92.7 % and a total sugar yield of 80 % from pretreated rice straw after 72 h of enzymatic hydrolysis.
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Aberrant proteomic expression of NSRP70 and its clinical implications and connection to the transcriptional level in adult acute leukemia.
Leuk. Res.
PUBLISHED: 08-10-2014
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We investigated three splicing factor proteins (SFPs; NSRP70, SRSF1, and HNRNPA1) in 187 adults with and without acute leukemia (AL). We showed that NSRP70 is a novel lymphoblastic AL (ALL) surrogate marker, which presented excellent diagnostic accuracy (92%) and disappeared during remission. Its highest molecular weight form, but not total amount, was associated with adverse genetic abnormalities in myeloid AL (AML). Furthermore, we identified that these SFPs were more prevalent in ALL than in AML; were not correlated with their mRNA levels; and their formations in AL may occur without coding mutations and relate to post-translational modifications.
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Promiscuous methionyl-tRNA synthetase mediates adaptive mistranslation against oxidative stresses.
J. Cell. Sci.
PUBLISHED: 08-07-2014
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Aminoacyl-tRNA Synthetases (ARSs) acylate tRNAs with amino acids. Charging tRNAs with the right amino acids is the first step in translation; therefore, the accurate and error-free functioning of ARSs is an essential prerequisite for translational fidelity. A recent study found that methionine (Met) can be incorporated into non-Met residues of proteins through methionylation to non-cognate tRNAs under oxidative stress. However, it was not understood how this mis-methionylation is achieved. Here, we report that methionyl-tRNA synthetase (MRS) is phosphorylated at Ser209 and Ser825 by extracellular signal-related kinase (ERK) upon reactive oxygen species (ROS) stress, and that this phosphorylated MRS showed increased affinity to non-cognate tRNAs with lower affinity to tRNA(Met), leading to an increase in Met residues in cellular proteins. The expression of a mutant MRS containing the substitutions S209D and S825D, mimicking dual phosphorylation, reduced ROS levels and cell death. This controlled inaccuracy of MRS seems to serve as a defense mechanism against ROS-mediated damage at the cost of translational fidelity.
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Heterozygous mutations in cyclic AMP phosphodiesterase-4D (PDE4D) and protein kinase A (PKA) provide new insights into the molecular pathology of acrodysostosis.
Cell. Signal.
PUBLISHED: 06-14-2014
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Acrodysostosis without hormone resistance is a rare skeletal disorder characterized by brachydactyly, nasal hypoplasia, mental retardation and occasionally developmental delay. Recently, loss-of-function mutations in the gene encoding cAMP-hydrolyzing phosphodiesterase-4D (PDE4D) have been reported to cause this rare condition but the pathomechanism has not been fully elucidated. To understand the pathogenetic mechanism of PDE4D mutations, we conducted 3D modeling studies to predict changes in the binding efficacy of cAMP to the catalytic pocket in PDE4D mutants. Our results indicated diminished enzyme activity in the two mutants we analyzed (Gly673Asp and Ile678Thr; based on PDE4D4 residue numbering). Ectopic expression of PDE4D mutants in HEK293 cells demonstrated this reduction in activity, which was identified by increased cAMP levels. However, the cells from an acrodysostosis patient showed low cAMP accumulation, which resulted in a decrease in the phosphorylated cAMP Response Element-Binding Protein (pCREB)/CREB ratio. The reason for this discrepancy was due to a compensatory increase in expression levels of PDE4A and PDE4B isoforms, which accounted for the paradoxical decrease in cAMP levels in the patient cells expressing mutant isoforms with a lowered PDE4D activity. Skeletal radiographs of 10-week-old knockout (KO) rats showed that the distal part of the forelimb was shorter than in wild-type (WT) rats and that all the metacarpals and phalanges were also shorter in KO, as the name acrodysostosis implies. Like the G-protein ?-stimulatory subunit and PRKAR1A, PDE4D critically regulates the cAMP signal transduction pathway and influences bone formation in a way that activity-compromising PDE4D mutations can result in skeletal dysplasia. We propose that specific inhibitory PDE4D mutations can lead to the molecular pathology of acrodysostosis without hormone resistance but that the pathological phenotype may well be dependent on an over-compensatory induction of other PDE4 isoforms that can be expected to be targeted to different signaling complexes and exert distinct effects on compartmentalized cAMP signaling.
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Delayed ischemic stroke after stent-assisted coil placement in cerebral aneurysm: characteristics and optimal duration of preventative dual antiplatelet therapy.
Radiology
PUBLISHED: 06-11-2014
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To evaluate characteristics of delayed ischemic stroke after stent-assisted coil placement in cerebral aneurysms and to determine the optimal duration of dual antiplatelet therapy for its prevention.
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Short-term change of handgrip strength after trigger point injection in women with muscular pain in the upper extremities.
Ann Rehabil Med
PUBLISHED: 04-29-2014
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To determine overall handgrip strength (HGS), we assessed the short-term change of HGS after trigger point injection (TPI) in women with muscular pain in the upper extremities by comparison with established pain scales.
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Optical and structural properties of plasma-treated ZnO nanostructures.
J Nanosci Nanotechnol
PUBLISHED: 04-25-2014
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We studied the effect of plasma treatment on the structural and optical properties of ZnO nanostructures prepared by chemical bath deposition in an aqueous solution of Zn(NO3)2 and hexamethylenetetramine. The room-temperature photoluminescence (PL) spectrum of the as-grown ZnO nanostructures exhibited two emission bands due to exciton emission and defect emission. After treating with hydrogen plasma, the treated ZnO nanostructures exhibited stronger exciton emission than the as-grown, untreated ZnO nanostructures in their respective cathodoluminescence and PL spectra. The low-temperature PL spectrum of the hydrogen plasma-treated ZnO nanostructures showed a strong exciton emission at 3.34 eV, attributing to the bound exciton and its longitudinal optical-phonon sidebands. The strong exciton emission is thought to be due to the combined effect of exciton emission enhancement by defect passivation and optical confinement resulting from nanostructure geometry.
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A biochemical and physicochemical comparison of two recombinant enzymes used for enzyme replacement therapies of hunter syndrome.
Glycoconj. J.
PUBLISHED: 04-07-2014
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Mucopolysaccharidosis II (MPS II, Hunter syndrome; OMIM 309900) is an X-linked lysosomal storage disease caused by a deficiency in the enzyme iduronate-2-sulfatase (IDS), leading to accumulation of glycosaminoglycans (GAGs). For enzyme replacement therapy (ERT) of Hunter syndrome, two recombinant enzymes, idursulfase (Elaprase(®), Shire Human Genetic Therapies, Lexington, MA) and idursulfase beta (Hunterase(®), Green Cross Corporation, Yongin, Korea), are currently available in Korea. To compare the biochemical and physicochemical differences between idursulfase and idursulfase beta, we examined the formylglycine (FGly) content, specific enzyme activity, mannose-6-phosphate (M6P) content, sialic acid content, and in vitro cell uptake activity of normal human fibroblasts of these two enzymes.The FGly content, which determines the enzyme activity, of idursulfase beta was significantly higher than that of idursulfase (79.4 ± 0.9 vs. 68.1 ± 2.2 %, P < 0.001). In accordance with the FGly content, the specific enzyme activity of idursulfase beta was significantly higher than that of idursulfase (42.6 ± 1.1 vs. 27.8 ± 0.9 nmol/min/?g protein, P < 0.001). The levels of M6P and sialic acid were not significantly different (2.4 ± 0.1 vs 2.4 ± 0.3 mol/mol protein for M6P and 12.3 ± 0.7 vs. 12.4 ± 0.4 mol/mol protein for sialic acid). However, the cellular uptake activity of the normal human fibroblasts in vitro showed a significant difference (Kuptake, 5.09 ± 0.96 vs. 6.50 ± 1.28 nM protein, P = 0.017).In conclusion, idursulfase beta exhibited significantly higher specific enzyme activity than idursulfase, resulting from higher FGly content. These biochemical differences may be partly attributed to clinical efficacy. However, long-term clinical evaluations of Hunter syndrome patients treated with these two enzymes will be needed to demonstrate the clinical implications of significant difference of the enzyme activity and the FGly content.
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Characterization of the interaction between lysyl-tRNA synthetase and laminin receptor by NMR.
FEBS Lett.
PUBLISHED: 04-04-2014
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Lysyl-tRNA synthetase (KRS) interacts with the laminin receptor (LR/RPSA) and enhances laminin-induced cell migration in cancer metastasis. In this nuclear magnetic resonance (NMR)-based study, we show that the anticodon-binding domain of KRS binds directly to the C-terminal region of 37LRP, and the previously found inhibitors BC-K-01 and BC-K-YH16899 interfere with KRS-37LRP binding. In addition, the anticodon-binding domain of KRS binds to laminin, observed by NMR and SPR. These results provide crucial insights into the structural characteristics of the KRS-LR interaction on the cell surface.
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The prevalence of idiopathic scoliosis in eleven year-old Korean adolescents: a 3 year epidemiological study.
Yonsei Med. J.
PUBLISHED: 04-01-2014
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School screening allows for early detection and early treatment of scoliosis, with the purpose of reducing the number of patients requiring surgical treatment. Children between 10 and 14 years old are considered as good candidates for school screening tests of scoliosis. The purpose of the present study was to assess the epidemiological findings of idiopathic scoliosis in 11-year-old Korean adolescents.
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MALDI-MS analysis and imaging of small molecule metabolites with 1,5-diaminonaphthalene (DAN).
J Mass Spectrom
PUBLISHED: 03-06-2014
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1,5-Diaminonaphthalene (DAN) has previously been reported as an effective matrix for matrix-assisted laser desorption ionization-mass spectrometry of phospholipids. In the current work, we investigate the use of DAN as a matrix for small metabolite analysis in negative ion mode. DAN was found to provide superior ionization to the compared matrices for MW?
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Potential masking of airway eosinophilic inflammation by combination therapy in asthma.
Allergy Asthma Immunol Res
PUBLISHED: 03-04-2014
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Long-acting ?2 agonists (LABA) may mask ongoing bronchial inflammation, leaving asthmatic patients at greater risk of severe complications. The aim of this study was to compare the effect of combination therapy using low-dose inhaled corticosteroids (ICS) plus LABA on airway inflammation in asthma to the effect of medium-dose ICS alone.
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Notch pathway activation is associated with pancreatic cancer treatment failure.
Pancreatology
PUBLISHED: 02-22-2014
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Pancreatic cancer is resistant to conventional treatment. The aim of the study was to confirm the hypothesis that changes in cancer stem cells (CSCs) and developmental pathway after treatment was responsible for treatment failure in pancreatic cancer.
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The effect of ketamine as an additive in epidural block on the intractable herpetic neuralgia: a case report.
Korean J Anesthesiol
PUBLISHED: 01-28-2014
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Ketamine has been shown to have analgesic effect by blocking N-methyl-D-aspartate receptor, thus preventing and reducing central sensitization caused by peripheral nociceptive stimulation. However, due to lack of knowledge about its safety and toxicity in the central nervous system, either epidural or intrathecal injection of ketamine still remains controversial. Here, we describe a case report of satisfactory pain relief after the addition of ketamine in epidural injection in a patient with severe herpes zoster pain that was refractory to conventional medication, intravenous opioids and continuous epidural block. This case indicates the viability of epidural ketamine injection in patients with intractable herpetic neuralgia.
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Development of a new method for the identification of degradation products of v-type nerve agents by liquid chromatography-tandem mass spectrometry.
J Chromatogr Sci
PUBLISHED: 01-26-2014
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Degradation products of V-type nerve agents are important markers of these toxic chemical warfare agents; hence their detection and identification are of high importance from verification point of view of Chemical Weapons Convention. The new analytical technique using quantitation-enhanced data-dependent (QED) method has been developed for the analysis of the degradation products, 2-(N,N-dialkylamino)ethanesulfonic acids of V-type nerve agents, by using liquid chromatography-tandem mass spectrometry (Thermo-Scientific Vantage triple stage quadrupole mass spectrometer, Thermo Finnigan Surveyor, San Jose, CA, USA) via an atmospheric pressure ionization source/interface operated in eletrospray ionization mode. With a single analytical run, we could perform the quantitative analysis of the 2-(N,N-dialkylamino)ethanesulfonic acids by the selected reaction monitoring scan mode with limit of detection at 0.1 ng/mL and identify their isomeric compounds by product ion scan mode, simultaneously. The QED method will be applicable to the trace analysis of degradation products of V-type nerve agents in the environmental matrices in the Organisation for the Prohibition of Chemical Weapons proficiency test.
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Analytical methods for chemical and sensory characterization of scent-markings in large wild mammals: a review.
Sensors (Basel)
PUBLISHED: 01-15-2014
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In conjoining the disciplines of "ethology" and "chemistry" the field of "Ethochemistry" has been instituted. Ethochemistry is an effective tool in conservation efforts of endangered species and the understanding of behavioral patterns across all species. Chemical constituents of scent-markings have an important, yet poorly understood function in territoriality, reproduction, dominance, and impact on evolutionary biology, especially in large mammals. Particular attention has recently been focused on scent-marking analysis of great cats (Kalahari leopards (Panthera pardus), puma (Puma concolor) snow leopard (Panthera uncia), African lions (Panthera leo), cheetahs (Acinonyx jubatus), and tigers (Panthera tigris)) for the purpose of conservation. Sensory analyses of scent-markings could address knowledge gaps in ethochemistry. The objective of this review is to summarize the current state-of-the art of both the chemical and sensory analyses of scent-markings in wild mammals. Specific focus is placed on sampling and sample preparation, chemical analysis, sensory analysis, and simultaneous chemical and sensory analyses. Constituents of exocrine and endocrine secretions have been most commonly studied with chromatography-based analytical separations. Odor analysis of scent-markings provides an insight into the animal's sensory perception. A limited number of articles have been published in the area of sensory characterization of scent marks. Simultaneous chemical and sensory analyses with chromatography-olfactometry hyphenation could potentially aid conservation efforts by linking perceived odor, compounds responsible for odor, and resulting behavior.
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A p38 MAPK-mediated alteration of COX-2/PGE2 regulates immunomodulatory properties in human mesenchymal stem cell aging.
PLoS ONE
PUBLISHED: 01-01-2014
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Because human mesenchymal stem cells (hMSC) have profound immunomodulatory effects, many attempts have been made to use hMSCs in preclinical and clinical trials. For hMSCs to be used in therapy, a large population of hMSCs must be generated by in vitro expansion. However, the immunomodulatory changes following the in vitro expansion of hMSCs have not been elucidated. In this study, we evaluated the effect of replicative senescence on the immunomodulatory ability of hMSCs in vitro and in vivo. Late-passage hMSCs showed impaired suppressive effect on mitogen-induced mononuclear cell proliferation. Strikingly, late-passage hMSCs had a significantly compromised protective effect against mouse experimental colitis, which was confirmed by gross and histologic examination. Among the anti-inflammatory cytokines, the production of prostaglandin E2 (PGE2) and the expression of its primary enzyme, cyclooxygenase-2 (COX-2), were profoundly increased by pre-stimulation with interferon gamma (IFN-?) and tumor necrosis factor alpha (TNF-?), and this response was significantly decreased with consecutive passages. We demonstrated that the impaired phosphorylation activity of p38 MAP kinase (p38 MAPK) in late-passage hMSCs led to a compromised immunomodulatory ability through the regulation of COX-2. In conclusion, our data indicate that the immunomodulatory ability of hMSCs gradually declines with consecutive passages via a p38-mediated alteration of COX-2 and PGE2 levels.
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Effects of two different doses of epidural steroid on blood glucose levels and pain control in patients with diabetes mellitus.
Pain Physician
PUBLISHED: 11-29-2013
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A high incidence of diabetes mellitus has been reported among patients diagnosed with lumbar degenerative spinal diseases. Although epidural steroid injections are known to increase the postprocedure blood glucose level, it has not been investigated whether a lower steroid dose can reduce blood glucose excursions and still be effective in controlling patients subjective pain.
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The role of second-look endoscopy in endoscopic submucosal dissection for early gastric cancer.
Turk J Gastroenterol
PUBLISHED: 11-15-2013
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Delayed bleeding is a major complication of endoscopic submucosal dissection. Second-look endoscopy is routinely performed in most hospitals to reduce the possibility of delayed bleeding without solid evidence to support this practice. The aim of this study was to evaluate whether second-look endoscopy prevents delayed bleeding, and to verify clinicopathological features of delayed bleeding in order to identify lesions that may benefit from a second-look endoscopy.
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Correlation between subfoveal choroidal thickness and the severity or progression of nonexudative age-related macular degeneration.
Invest. Ophthalmol. Vis. Sci.
PUBLISHED: 11-09-2013
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To investigate the correlation between subfoveal choroidal thickness (SFChT) and the severity or progression of nonexudative AMD.
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Surgical correction of epiblepharon using an epicanthal weakening procedure with lash rotating sutures.
Br J Ophthalmol
PUBLISHED: 10-17-2013
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To describe a new surgical technique in patients with lower eyelid epiblepharon using an epicanthal weakening procedure with lash rotating sutures.
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Intracranial Stenting Using a Drug-Eluting Stent for Moyamoya Disease Involving Supraclinoid ICA: A Case Report.
Neurol. Med. Chir. (Tokyo)
PUBLISHED: 10-07-2013
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A 43-year-old woman was diagnosed with moyamoya disease (MMD) and underwent right-side bypass surgery. After surgery, previous symptoms disappeared. One month later, transient right hemiparetic attacks and motor dysphasia developed. Angiography revealed progressive severe stenosis of left supraclinoid segment of internal carotid artery. Angioplasty using a drug-eluting stent (DES) was performed. For 18 months, she presented no ischemic symptom and no instent stenosis was observed in follow-up angiography. This is the first case report about effect of DES use for MMD. Considering that intimal hyperplasia is a pathophysiology of stenosis, DES may have a role in reducing progression of stenosis in selected moyamoya patients.
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Different Zn(II) cation coordination geometries in di-?-acetato-bis{2-chloro-6-[(pyridine-2-ylmethylimino)methyl]phenol}dizinc(II) chloroform monosolvate.
Acta Crystallogr C
PUBLISHED: 08-17-2013
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In the title compound, di-?-acetato-?(2)O:O;?(2)O:O-bis[(6-chloro-2-{(E)-[(pyridin-2-yl)methylimino]methyl}phenolato-?(3)N,N,O)zinc(II)], [Zn2(C13H10ClN2O)2(C2H3O2)2]·CHCl3, the Zn(II) cation adopts a five-coordinate geometry and is coordinated by two N atoms and one O atom of a tridentate 6-chloro-2-{(E)-[(pyridin-2-yl)methylimino]methyl}phenolate ligand and two O atoms of two bridging acetate groups, but their coordination geometries differ. One Zn(II) cation adopts a distorted trigonal bipyramidal geometry and the other a square-pyramidal geometry. The two acetate ligands bridge two Zn(II) cations with mono- and bidentate coordination modes. The title compound exhibits a strong emission at 460?nm upon excitation at 325?nm with a quantum yield of 23.1%.
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Levetiracetam compared with valproic acid for the prevention of postoperative seizures after supratentorial tumor surgery: a retrospective chart review.
CNS Drugs
PUBLISHED: 08-08-2013
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Antiepileptic drugs are commonly given for perioperative prophylaxis after brain tumor surgery, and there has been growing interest in levetiracetam, a second-generation antiepileptic drug. This retrospective study compared the seizure outcomes, side effects and durability of levetiracetam with valproic acid after a craniotomy for supratentorial brain tumors.
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Chemical suppression of an oncogenic splicing variant of AIMP2 induces tumour regression.
Biochem. J.
PUBLISHED: 07-03-2013
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AIMP2 (aminoacyl-tRNA synthetase-interacting multifunctional protein 2) is a potent tumour suppressor that induces apoptosis in response to various oncogenic signals. AIMP2-DX2, an exon2-deleted splicing variant of AIMP2, is up-regulated in lung cancer and competitively suppresses the pro-apoptotic activity of AIMP2, resulting in tumorigenesis. In the present study we report that BC-DXI01, a synthetic compound, specifically reduces the cellular levels of AIMP2-DX2 through selective degradation of the AIMP2-DX2 mRNA transcript. We found that BC-DXI01-mediated cell death positively correlates with AIMP2-DX2 expression in the lung cancer cell lines tested. Administration of BC-DXI01 in a AIMP2-DX2-driven tumour xenograft mice model led to reduced tumour sizes and volumes of up to 60% in comparison with vehicle-treated mice group, consistent with decreases in AIMP2-DX2 transcript and protein levels. Taken together, our findings suggest that tumorigenic activity of AIMP2-DX2 can be controlled by the small chemical BC-DXI01, which can selectively suppress the AIMP2-DX2 mRNA transcript.
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Chemical inhibition of prometastatic lysyl-tRNA synthetase-laminin receptor interaction.
Nat. Chem. Biol.
PUBLISHED: 06-18-2013
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Lysyl-tRNA synthetase (KRS), a protein synthesis enzyme in the cytosol, relocates to the plasma membrane after a laminin signal and stabilizes a 67-kDa laminin receptor (67LR) that is implicated in cancer metastasis; however, its potential as an antimetastatic therapeutic target has not been explored. We found that the small compound BC-K-YH16899, which binds KRS, impinged on the interaction of KRS with 67LR and suppressed metastasis in three different mouse models. The compound inhibited the KRS-67LR interaction in two ways. First, it directly blocked the association between KRS and 67LR. Second, it suppressed the dynamic movement of the N-terminal extension of KRS and reduced membrane localization of KRS. However, it did not affect the catalytic activity of KRS. Our results suggest that specific modulation of a cancer-related KRS-67LR interaction may offer a way to control metastasis while avoiding the toxicities associated with inhibition of the normal functions of KRS.
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Copper(L) selenide thin films deposited by a solution-based method for photovoltaic applications.
J Nanosci Nanotechnol
PUBLISHED: 06-13-2013
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Polycrystalline Cu(2-x)Se thin films for solar cell application were deposited by a solution-based continuous flow reactor (CFR) process. In order to study the influence of reaction time on the physical, structural, and optical properties of the Cu(2-x)Se thin films, the deposition time for impinging varied from 2 min to 15 min. Based on X-ray diffraction (XRD), scanning electron microscopy (SEM), and UV-visible spectrophotometry, the optimum time to deposit the Cu(2-x)Se thin films using the CFR process was 10 min. The X-ray diffraction analysis revealed the cubic structure of the Cu(2-x)Se thin films annealed at 400 degrees C. The estimated band gap of the film was approximately 2.20 eV and its average grain size was around 150 nm with a film thickness of 1.8 microm. Energy dispersive analysis by X-ray (EDAX) showed that the atomic weight ratio of Cu to Se in the Cu(2-x)Se film deposited for 10 min was 1.8:1, i.e., x = 0.2. The chemical binding information of the Cu(2-x)Se thin film was also studied using X-ray photoelectron spectroscopy (XPS).
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A simple bacterial transformation method using magnesium- and calcium-aminoclays.
J. Microbiol. Methods
PUBLISHED: 06-12-2013
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An efficient and user-friendly bacterial transformation method by simple spreading cells with aminoclays was demonstrated. Compared to the reported transformation approaches using DNA adsorption or wrapping onto (in)organic fibers, the spontaneously generated clay-coated DNA suprastructures by mixing DNA with aminoclay resulted in transformants in both Gram-negative (Escherichia coli) and Gram-positive cells (Streptococcus mutans). Notably, the wild type S. mutans showed comparable transformation efficiency to that of the E. coli host for recombinant DNA cloning. This is a potentially promising result because other trials such as heat-shock, electroporation, and treatment with sepiolite for introducing DNA into the wild type S. mutans failed. Under defined conditions, the transformation efficiency of E. coli XL1-Blue and S. mutans exhibited ~2 × 10(5) and ~6 × 10(3)CFU/?g of plasmid DNA using magnesium-aminoclay. In contrast, transformation efficiency was higher in S. mutans than that in E. coli XL1-Blue for calcium-aminoclay. It was also confirmed that each plasmid transformed into E. coli and S. mutans was stably maintained and that they expressed the inserted gene encoding the green fluorescent protein during prolonged growth of up to 80 generations.
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Sex-specific expression of CTNNB1 in the gonadal morphogenesis of the chicken.
Reprod. Biol. Endocrinol.
PUBLISHED: 05-31-2013
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Beta-catenin (CTNNB1), as a key transcriptional regulator in the WNT signal transduction cascade, plays a pivotal role in multiple biological functions such as embryonic development and homeostasis in adults. Although it has been suggested that CTNNB1 is required for gonad development and maintenance of ovarian function in mice, little is known about the expression and functional role of CTNNB1 in gonadal development and differentiation in the chicken reproductive system.
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Changes in prion replication environment cause prion strain mutation.
FASEB J.
PUBLISHED: 05-31-2013
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Interspecies prion transmission often leads to stable changes in physical and biological features of prion strains, a phenomenon referred to as a strain mutation. It remains unknown whether changes in the replication environment in the absence of changes in PrP primary structure can be a source of strain mutations. To approach this question, RNA content was altered in the course of amplification of hamster strains in serial protein misfolding cyclic amplification (sPMCAb). On adaptation to an RNA-depleted environment and then readaptation to an environment containing RNA, strain 263K gave rise to a novel PrP(Sc) conformation referred to as 263K(R+), which is characterized by very low conformational stability, high sensitivity to proteolytic digestion, and a replication rate of 10(6)-fold/PMCAb round, which exceeded that of 263K by almost 10(4)-fold. A series of PMCAb experiments revealed that 263K(R+) was lacking in brain-derived 263K material, but emerged de novo as a result of changes in RNA content. A similar transformation was also observed for strain Hyper, suggesting that this phenomenon was not limited to 263K. The current work demonstrates that dramatic PrP(Sc) transformations can be induced by changes in the prion replication environment and without changes in PrP primary structure.
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Expression analysis and enzymatic characterization of phospholipase C?4 from olive flounder (Paralichthys olivaceus).
Comp. Biochem. Physiol. B, Biochem. Mol. Biol.
PUBLISHED: 05-20-2013
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Phospholipase C?4 (PLC?4) plays a significant role in cell proliferation, tumorigenesis, and in an early stage of fertilization. Despite the characterization of the mammalian PLC?4, extensive study in aquatic organisms has not been carried out so far. Here, we performed the molecular and biochemical characterization of flatfish Paralichthys olivaceus PLC?4 (PoPLC?4) to understand its enzymatic properties and physiological functions. The olive flounder PLC?4 cDNA has an open reading frame (ORF) of 2,268 bp, and encodes a 755 amino acid polypeptide with a predicted molecular weight of 86 kDa. All the characteristic domains found in mammalian PLC? isoforms (PH domain, EF hands, an X-Y catalytic region, and a C2 domain) were found to be present in PoPLC?4. The mRNA expression analysis of PoPLC?4 showed that PoPLC?4 is predominantly expressed in the brain, eye and heart tissues. Like other mammalian PLC? proteins, the enzyme activity of recombinant PoPLC?4 to phosphatidylinositol-4,5-bis-phosphate (PIP2) was noted to be concentration- and Ca(2+)-dependent. The structural features and biochemical characteristics of PoPLC?4 were found to be similar to those of mammalian PLC?4. This is the first demonstration of the expression analysis and enzymatic characterization of piscine PLC?4.
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Hydrogen-atom-mediated electrochemistry.
Nat Commun
PUBLISHED: 05-19-2013
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Silicon dioxide thin films are widely used as dielectric layers in microelectronics and can also be engineered on silicon wafers. It seems counterintuitive that electrochemical reactions could occur on such an insulator without relying on tunnelling current. Here we report electrochemistry based on electron transfer through a thin insulating layer of thermally grown silicon dioxide on highly n-doped silicon. Under a negative electrical bias, protons in the silicon dioxide layer were reduced to hydrogen atoms, which served as electron mediators for electrochemical reduction. Palladium nanoparticles were preferentially formed on the dielectric layer and enabled another hydrogen-atom-mediated electrochemistry, as their surfaces retained many electrogenerated hydrogen atoms to act as a hydrogen-atom reservoir for subsequent electrochemical reduction. By harnessing the precisely controlled electrochemical generation of hydrogen atoms, palladium-copper nanocrystals were synthesized without any surfactant or stabilizer on the silicon dioxide layer.
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Olive flounder (Paralichthys olivaceus) cystatin B: cloning, tissue distribution, expression and inhibitory profile of piscine cystatin B.
Comp. Biochem. Physiol. B, Biochem. Mol. Biol.
PUBLISHED: 04-29-2013
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Among the cystatin superfamily, cystatin B, also known as stefin B, is an intracellular inhibitor that regulates the activities of cysteine proteases, such as papain and cathepsins. In this study, the 536 bp cystatin B cDNA (referred to hereafter as PoCystatin B) was cloned from olive flounder (Paralichthys olivaceus) using a combination of the rapid amplification of cDNA ends (RACE) approach and olive flounder cDNA library screening. To determine the tissue distribution of PoCystatin B mRNA, the expression of PoCystatin B in normal and lipopolysaccharide (LPS)-stimulated flounder tissues were compared with that of the inflammatory cytokines interleukin (IL)-1?, IL-6, and IL-8 by reverse transcription (RT)-polymerase chain reaction (PCR). The results of the RT-PCR analysis revealed ubiquitous PoCystatin B expression in normal and LPS-stimulated tissues. To characterize the enzymatic activity of PoCystatin B protein, recombinant PoCystatin B protein was overexpressed in Escherichia coli BL21(DE3) cells in the pCold™ TF DNA expression vector as a soluble fusion protein of 67-kDa. PoCystatin B inhibited papain cysteine protease, bovine cathepsin B, and fish cathepsins F and X to a greater extent, whereas fish cathepsins L, S, and K were inhibited to a lesser extent. These results indicate that the enzymatic characteristics of the olive flounder cystatin B are similar to those of mammalian cystatin B proteins, and provide a better understanding of the mechanisms of regulation of cathepsins and cystatins in marine organisms.
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Identification of sensory attributes that drive consumer liking of commercial orange juice products in Korea.
J. Food Sci.
PUBLISHED: 04-14-2013
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Orange juice is a well-accepted fruit juice, and its consumption increases steadily. Many studies have been conducted to understand the sensory characteristics of orange juice throughout its varying processing steps. Sensory language and consumer likings of food can be influenced by culture. The objective of this study is to evaluate the sensory characteristics of commercially available orange juices in Korea and identify drivers of liking for orange juices in Korea. A quantitative descriptive analysis was conducted using a trained panel (n = 10) to evaluate 7 orange juice samples in triplicates, followed by consumer acceptance tests (n = 103). Univariate and multivariate statistical analyses were conducted for data analysis. The sensory characteristics of commercially available orange juice were documented and grouped: group 1 samples were characterized by high in natural citrus flavors such as orange peel, orange flesh, citrus fruit, and grape fruit, whereas group 2 samples were characterized by processed orange-like flavors such as over-ripe, cooked-orange, and yogurt. Regardless of orange flavor types, a high intensity of orange flavor in orange juice was identified as a driver of liking for orange juices in Korea. Three distinct clusters were segmented by varying sensory attributes that were evaluated by likes and dislikes. Overall, many similarities were noticed between Korean market segment and global orange juice market. By knowing the drivers of liking and understanding the distinct consumer clusters present in the Korean orange juice market, the orange juice industry could improve the strategic marketing of its products in Korea.
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Case series of an intraoral balancing appliance therapy on subjective symptom severity and cervical spine alignment.
Evid Based Complement Alternat Med
PUBLISHED: 04-08-2013
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Objective. The objective of this study was to investigate the effect of a holistic intraoral appliance (OA) on cervical spine alignment and subjective symptom severity. Design. An observational study on case series with holistic OA therapy. Setting. An outpatient clinic for holistic temporomandibular joint (TMJ) therapy under the supervision of the Pain Center, CHA Biomedical center, CHA University. Subjects. Ambulatory patients presenting with diverse chief complaints in the holistic TMJ clinic. Main Measures. Any immediate change in the curvature of cervical spine and the degree of atlantoaxial rotation was investigated in the images of simple X-ray and computed tomography of cervical spine with or without OA. Changes of subjective symptom severity were also analyzed for the holistic OA therapy cases. Results. A total of 59 cases were reviewed. Alignment of upper cervical spine rotation showed an immediate improvement (P < 0.001). Changes of subjective symptom severity also showed significant improvement (P < 0.05). Conclusion. These cases revealed rudimentary clinical evidence that holistic OA therapy may be related to an alleviated symptom severity and an improved cervical spinal alignment. These results show that further researches may warrant for the holistic TMJ therapy.
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Primary extraskeletal mesenchymal chondrosarcoma of the vulva.
Obstet Gynecol Sci
PUBLISHED: 04-05-2013
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Extraskeletal chondrosarcoma is rare, making up only 1% of reported chondrosarcoma. We experienced 3 cases of extraskeletal chondrosarcoma, especially in vulva. They were suspected as lipoma of the vulva. The patients had noticed a small but growing mass on their vulva which had been palpated earlier. The masses were excised with a 2 cm resection margin. The final pathological reports confirmed extraskeletal mesenchymal chondrosarcoma (EMC) of the vulva revealing no microscopic lesions on the resection margins. After 24 months of following from the initial diagnosis, the patients remain without evidence of any recurrent. Management of EMC is not well studied due to the rare and variable nature of the disease. However, the surgery, such as we had, is the mainstay of local treatment with studies showing better survival in patients who undergo wide surgical resection. The establishment of adjuvant systemic pharmacotherapy could be expected in the future.
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Loss of lordosis and clinical outcomes after anterior cervical fusion with dynamic rotational plates.
Yonsei Med. J.
PUBLISHED: 04-04-2013
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The cervical dynamic rotational plating system may induce bone graft subsidence, so it may cause loss of cervical lordosis. However there were few studies for alignments of cervical spines influencing the clinical results after using dynamic rotational plates. The purpose is to evaluate the effect of graft subsidence on cervical alignments due to the dynamic rotational cervical plates and correlating it with the clinical outcomes of patients undergoing anterior cervical fusion.
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Hormonal regulation of beta-catenin during development of the avian oviduct and its expression in epithelial cell-derived ovarian carcinogenesis.
Mol. Cell. Endocrinol.
PUBLISHED: 03-18-2013
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Beta-catenin (CTNNB1) is a dual function molecule that acts as a key component of the cadherin complex and WNT signaling pathway. It has a crucial role in embryogenesis, tumorigenesis, angiogenesis and progression of metastasis. Recently, it has been suggested that the CTNNB1 complex is a major regulator of development of the mouse oviduct and uterus. However, little is known about the CTNNB1 gene in chickens. Therefore, in this study, we focused on the CTNNB1 gene in the chicken reproductive tract and hormonal control of its expression in the chicken oviduct. CTNNB1 was localized specifically to the luminal and glandular epithelium of the four segments of chicken oviduct and DES (diethylstilbestrol, a synthetic non-steroidal estrogen) increased its expression primarily in LE of the magnum. In addition, CTNNB1 mRNA and protein were expressed abundantly in glandular epithelium of endometrioid-type ovarian carcinoma, but not in normal ovaries. Moreover, CTNNB1 expression was post-transcriptionally regulated via its 3-UTR by binding with target miRNAs including miR-217, miR-1467, miR-1623 and miR-1697. Collectively, these results indicate that CTNNB1 is a novel gene regulated by estrogen in epithelial cells of the chicken oviduct and that it is also abundantly expressed in epithelial cells of endometrioid-type ovarian carcinoma suggesting that it could be used as a marker for diagnosis of ovarian cancer in laying hens and women.
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SPARC is involved in the maintenance of mitotically inactivated mouse embryonic fibroblast cells.
In Vitro Cell. Dev. Biol. Anim.
PUBLISHED: 03-13-2013
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Mitotically inactivated feeder cells such as mouse embryonic fibroblast (MEFs) cells have been widely applied for physical and physiological support in the pluripotency maintenance of human pluripotent stem cells (hPSCs). However, accurate supporting mechanism or factors of feeder cells are poorly understood. Here, we isolated differentially expressed genes between wild-type MEFs and mitotically inactivated MEFs (miMEFs) by employing annealing control primer-based GeneFishing polymerase chain reaction. We identified a secreted protein acidic cysteine-rich glycoprotein (SPARC) gene that is upregulated in miMEFs. Suppression of SPARC expression in miMEFs using small interference RNA (siRNA) displayed gradual detachment of miMEFs. Furthermore, we found a significant reduction of OCT4- and SSEA3-positive hPS cell population maintained on SPARC siRNA-miMEFs compared to on miMEFs by flow cytometrical analysis. These findings suggest that SPARC plays a critical role in the maintenance of miMEFs without loss of cell number and might be a key component for supporting the culture of hPSCs.
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Multiplex mass spectrometric imaging with polarity switching for concurrent acquisition of positive and negative ion images.
J. Am. Soc. Mass Spectrom.
PUBLISHED: 03-07-2013
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We have recently developed a multiplex mass spectrometry imaging (MSI) method which incorporates high mass resolution imaging and MS/MS and MS(3) imaging of several compounds in a single data acquisition utilizing a hybrid linear ion trap-Orbitrap mass spectrometer (Perdian and Lee, Anal. Chem. 82, 9393-9400, 2010). Here we extend this capability to obtain positive and negative ion MS and MS/MS spectra in a single MS imaging experiment through polarity switching within spiral steps of each raster step. This methodology was demonstrated for the analysis of various lipid class compounds in a section of mouse brain. This allows for simultaneous imaging of compounds that are readily ionized in positive mode (e.g., phosphatidylcholines and sphingomyelins) and those that are readily ionized in negative mode (e.g., sulfatides, phosphatidylinositols and phosphatidylserines). MS/MS imaging was also performed for a few compounds in both positive and negative ion mode within the same experimental set-up. Insufficient stabilization time for the Orbitrap high voltage leads to slight deviations in observed masses, but these deviations are systematic and were easily corrected with a two-point calibration to background ions.
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Progression of retinal nerve fiber layer thinning in glaucoma assessed by cirrus optical coherence tomography-guided progression analysis.
Curr. Eye Res.
PUBLISHED: 02-28-2013
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To evaluate the performance of Cirrus spectral domain optical coherence tomography (SD-OCT)-guided progression analysis (GPA) software to detect progression of retinal nerve fiber layer (RNFL) thinning in glaucoma patients.
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Clinical characteristics of glaucomatous subjects treated with refractive corneal ablation surgery.
Korean J Ophthalmol
PUBLISHED: 02-27-2013
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To evaluate the clinical characteristics of newly diagnosed glaucomatous subjects who had a history of refractive corneal ablation surgery (RCAS).
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Recurrent cerebral infarction caused by mobile aortic arch thrombus refractory to warfarin therapy.
Ann Rehabil Med
PUBLISHED: 02-26-2013
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The mobile thrombus in the aortic arch is a rare condition, which becomes rarer when associated with cerebral infarction, resulting in disabling complications. Transesophageal echocardiography is useful for detecting the source of thrombi in the heart and aortic arch. Here, we report a case of a patient who suffered from recurrent cerebral infarction four times during the previous 18 months due to mobile thrombus in the aortic arch despite anticoagulant therapy. The thrombus with rocking motion (9.6 and 8 mm) was laid in the top of the aortic arch, and surgical resection was not allowed due to the location. We began anticoagulation therapy initially with heparin followed by a combination of warfarin, aspirin, and atorvastatin with an international normalized ratio between 2 and 3. The size of the aortic thrombi was significantly decreased by 3 mm and stabilized after 18 months.
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Two-year results of AcrySof toric intraocular lens implantation in patients with combined microincision vitrectomy surgery and phacoemulsification.
Br J Ophthalmol
PUBLISHED: 02-01-2013
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To evaluate the effects and stability of AcrySof toric intraocular lens (IOL) implantation in patients who had combined microincision vitrectomy surgery (MIVS) and phacoemulsification for vitreoretinal diseases and cataract with corneal astigmatism.
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Higher Prevalence of Klebsiella pneumoniae Extended-Spectrum ?-Lactamase in Patients on Renal Replacement Therapy.
J. Korean Med. Sci.
PUBLISHED: 01-24-2013
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The prevalence of antibiotic resistance is higher in patients undergoing renal replacement therapy (RRT) than in patients who did not undergo RRT. We investigated the presence of KP (Klebsiella pneumoniae) in patients who underwent RRT. All data were collected retrospectively by accessing patient medical records from 2004 to 2011 for the culture results of all patients who were positive for KP. We grouped the patients by the presence of extended-spectrum ?-lactamase (ESBL) into a KP ESBL(-) group (KP[-]) and a KP ESBL(+) group (KP[+]). In total, 292 patients (23.1%) were in the KP(+) group, and 974 patients (76.9%) were in the KP(-) group. A greater percentage of KP(+) was found in patients who underwent RRT (7.5%) than in patients who did not undergo RRT (3.2%) (OR, 2.479; 95% CI,1.412-4.352). A Coxs hazard proportional model analysis was performed, and for patients with pneumonia, the risk of KP(+) was 0.663 times higher in patients who had lower albumin levels, 2.796 times higher in patients who had an inserted Levin tube, and 4.551 times higher in patients who underwent RRT. In conclusion, RRT can be a risk factor for KP(+) in patients with pneumonia.
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Inhibition of SCAMP1 suppresses cell migration and invasion in human pancreatic and gallbladder cancer cells.
Tumour Biol.
PUBLISHED: 01-23-2013
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Lymph node (LN) metastasis is one of the most important risk factors for the prognosis of pancreatic cancer. This study aimed to identify novel LN metastasis-associated markers and therapeutic targets for pancreatic and gallbladder cancers. DNA microarray analysis was carried out to identify genes differentially expressed between 17 pancreatic cancer tissues with LN metastasis and 17 pancreatic cancer tissues without LN metastasis. The expression of LZIC, FXR, SCAMP1, and SULT1E1 is significantly higher in pancreatic cancer tissues with LN metastasis than in pancreatic cancer tissues without LN metastasis. We recently reported that FXR plays an important role in LN metastasis of pancreatic cancer, and in this study, we selected the secretory carrier membrane protein 1 (SCAMP1) gene. To determine that function of the SCAMP1 gene, we examined the effects of SCAMP1 knockdown on pancreatic and gallbladder cancer proliferation, migration, and invasion using SCAMP1 small interfering RNA (siRNA) and the activity of vascular endothelial growth factor (VEGF) was measured by enzyme-linked immunosorbent assay. SCAMP1 overexpression is associated with LN metastasis in pancreatic cancer patients. The siRNA-mediated downregulation of SCAMP1 resulted in a marked reduction in cell migration and invasion, but not proliferation in MIA-PaCa2, PANC-1, TGBC-1, and TGBC-2 cells. In addition, downregulation of SCAMP1 inhibited VEGF levels of conditioned medium from SCAMP1 siRNA-transfected cells. These results suggest that downregulation of SCAMP1 could be a potential therapeutic target for patients with pancreatic and gallbladder cancer.
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Remarkable reinforcement of a supramolecular gel constructed by heteroditopic [18]crown-6-based molecular recognition.
Chemistry
PUBLISHED: 01-16-2013
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The heteroditopic crown ether-based ligand 1 containing the diazafluorenylimino group as a binding site for the Zn(2+) and Cs(+) was synthesized. The ligand 1 can be gelated in DMSO/water with and without Cs(+) in the presence of the Zn(2+) ion. Interestingly, the remarkable gelation reinforcement of gel 1 occurred with Cs(+) in the presence of Zn(2+), which is due to the formation of a sandwich complex. According to DFT calculations, one Zn(2+) is bound to two diazafluorenylimino moieties in a tetrahedral structure. In addition, one Cs(+) ion is bound to two crown-rings. The Zn(2+)-diazafluorenylimino ligand gel without Cs(+) shows a spherical structure with 250-800 nm diameter, whereas the Zn(2+)-diazafluorenylimino-based ligand gel with Cs(+) shows a fiber structure with 60-70 nm diameter and several micrometers of lengths. The rheological properties of the Zn(2+)-diazafluorenylimino-based ligand gel were strongly dependent on the presence of Cs(+).
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PLC-?1-Lf, a novel N-terminal extended phospholipase C-?1.
Gene
PUBLISHED: 01-11-2013
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Phospholipase C-? (PLC-?), a key enzyme in phosphoinositide turnover, is involved in a variety of physiological functions. The widely expressed PLC-?1 isoform is the best characterized and the most well understood phospholipase family member. However, the functional and molecular mechanisms of PLC-?1 remain obscure. Here, we identified that the N-terminal region of mouse PLC-?1 gene has two variants, a novel alternative splicing form, named as long form (mPLC-?1-Lf) and the previously reported short form (mPLC-?1-Sf), having exon 2 and exon 1, respectively, while both the gene variants share exons 3-16 for RNA transcription. Furthermore, the expression, identification and enzymatic characterization of the two types of PLC-?1 genes were compared. Expression of mPLC-?1-Lf was found to be tissue specific, whereas mPLC-?1-Sf was widely distributed. The recombinant mPLC-?1-Sf protein exhibited higher activity than recombinant mPLC-?1-Lf protein. Although, the general catalytic and regulatory properties of mPLC-?1-Lf are similar to those of PLC-?1-Sf isozyme, the mPLC-?1-Lf showed some distinct regulatory properties, such as tissue-specific expression and lipid binding specificity, particularly for phosphatidylserine.
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Fate of newly detected lesions during postoperative surveillance for non-small cell lung cancer.
Ann. Thorac. Surg.
PUBLISHED: 01-11-2013
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Chest computed tomography (CT) is the mainstay of postoperative surveillance for non-small cell lung cancer (NSCLC). However, there is no clear consensus about the optimal management of newly detected lesions on follow-up CT. Our goals were (1) to determine the eventual outcome of newly detected lesions on follow-up CT in patients with previously resected NSCLC and (2) to determine the characteristics of the detected lesions that suggest recurrence.
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Multiplex mass spectrometry imaging for latent fingerprints.
J Mass Spectrom
PUBLISHED: 01-11-2013
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We have previously developed in-parallel data acquisition of orbitrap mass spectrometry (MS) and ion trap MS and/or MS/MS scans for matrix-assisted laser desorption/ionization MS imaging (MSI) to obtain rich chemical information in less data acquisition time. In the present study, we demonstrate a novel application of this multiplex MSI methodology for latent fingerprints. In a single imaging experiment, we could obtain chemical images of various endogenous and exogenous compounds, along with simultaneous MS/MS images of a few selected compounds. This work confirms the usefulness of multiplex MSI to explore chemical markers when the sample specimen is very limited.
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Olive flounder (Paralichthys olivaceus) cystatin C: cloning, mRNA expression, and enzymatic characterization of olive flounder cystatin C.
Appl. Biochem. Biotechnol.
PUBLISHED: 01-09-2013
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Cystatins are endogenous inhibitors of mammalian lysosomal cysteine proteinases, such as cathepsins B, L, H, and S. Cystatin C belongs to the type 2 cystatin family. In this study, the 751-bp cystatin C cDNA (PoCystatin C) of olive flounder (Paralichthys olivaceus) was cloned by screening from the olive flounder cDNA library. The mRNA expression of the PoCystatin C gene was examined in various tissues from normal and lipopolysaccharide (LPS)-stimulated olive flounder by RT-PCR and was compared with inflammatory cytokines IL-1?, IL-6, and IL-8. PoCystatin C transcripts ubiquitously existed in all normal and LPS-stimulated tissues that were tested. The recombinant PoCystatin C protein was expressed in Escherichia coli BL21(DE3) in pCold™ TF DNA expression vector as a 70-kDa fusion protein. The protease inhibitory activities of recombinant PoCystatin C toward papain cysteine protease, piscine cathepsins (L, S, K, F, and X), and bovine cathepsin B were measured with the synthetic fluorogenic peptide substrates. PoCystatin C tightly inhibited papain cysteine protease, whereas cathepsins L, S, K, F, X, and B were inhibited with lower affinities. Our results indicate that the P. olivaceus cystatin C is a homolog of mammalian cystatin C due to its sequence, structure, tissue expression, and biochemical activity.
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High-throughput analysis of algal crude oils using high resolution mass spectrometry.
Lipids
PUBLISHED: 01-03-2013
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Lipid analysis often needs to be specifically optimized for each class of compounds due to its wide variety of chemical and physical properties. It becomes a serious bottleneck in the development of algae-based next generation biofuels when high-throughput analysis becomes essential for the optimization of various process conditions. We propose a high-resolution mass spectrometry-based high-throughput assay as a quick-and-dirty protocol to monitor various lipid classes in algal crude oils. Atmospheric pressure chemical ionization was determined to be most effective for this purpose to cover a wide range of lipid classes. With an autosampler-LC pump set-up, we could analyze algal crude samples every one and half minutes, monitoring several lipid species such as TAG, DAG, squalene, sterols, and chlorophyll a. High-mass resolution and high-mass accuracy of the orbitrap mass analyzer provides confidence in the identification of these lipid compounds. MS/MS and MS3 analysis could be performed in parallel for further structural information, as demonstrated for TAG and DAG. This high-throughput method was successfully demonstrated for semi-quantitative analysis of algal oils after treatment with various nanoparticles.
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Recrudescence mechanisms and gene expression profile of the reproductive tracts from chickens during the molting period.
PLoS ONE
PUBLISHED: 01-01-2013
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The reproductive system of chickens undergoes dynamic morphological and functional tissue remodeling during the molting period. The present study identified global gene expression profiles following oviductal tissue regression and regeneration in laying hens in which molting was induced by feeding high levels of zinc in the diet. During the molting and recrudescence processes, progressive morphological and physiological changes included regression and re-growth of reproductive organs and fluctuations in concentrations of testosterone, progesterone, estradiol and corticosterone in blood. The cDNA microarray analysis of oviductal tissues revealed the biological significance of gene expression-based modulation in oviductal tissue during its remodeling. Based on the gene expression profiles, expression patterns of selected genes such as, TF, ANGPTL3, p20K, PTN, AvBD11 and SERPINB3 exhibited similar patterns in expression with gradual decreases during regression of the oviduct and sequential increases during resurrection of the functional oviduct. Also, miR-1689* inhibited expression of Sp1, while miR-17-3p, miR-22* and miR-1764 inhibited expression of STAT1. Similarly, chicken miR-1562 and miR-138 reduced the expression of ANGPTL3 and p20K, respectively. These results suggest that these differentially regulated genes are closely correlated with the molecular mechanism(s) for development and tissue remodeling of the avian female reproductive tract, and that miRNA-mediated regulation of key genes likely contributes to remodeling of the avian reproductive tract by controlling expression of those genes post-transcriptionally. The discovered global gene profiles provide new molecular candidates responsible for regulating morphological and functional recrudescence of the avian reproductive tract, and provide novel insights into understanding the remodeling process at the genomic and epigenomic levels.
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Avian WNT4 in the female reproductive tracts: potential role of oviduct development and ovarian carcinogenesis.
PLoS ONE
PUBLISHED: 01-01-2013
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The wingless-type MMTV integration site family of proteins (WNTs) is highly conserved secreted lipid-modified signaling molecules that play a variety of pivotal roles in developmental events such as embryogenesis, tissue homeostasis and cell polarity. Although, of these proteins, WNT4 is known to be involved in genital development in fetuses of mammalian species, its role is unknown in avian species. Therefore, in this study, we investigated expression profiles, as well as hormonal and post-transcriptional regulation of WNT4 expression in the reproductive tract of female chickens. Results of this study demonstrated that WNT4 is most abundant in the stromal and luminal epithelial cells of the isthmus and shell gland of the oviduct, respectively. WNT4 is also most abundant in the glandular epithelium of the shell gland of the oviduct of laying hens at 3 h post-ovulation during the laying cycle. In addition, treatment of young chicks with diethylstilbestrol (DES, a synthetic estrogen agonist) stimulated WNT4 only in the glandular epithelial cells of the isthmus and shell gland of the oviduct. Moreover, results of our study demonstrated that miR-1786 influences WNT4 expression via specific binding sites in its 3-UTR. On the other hand, our results also indicate that WNT4 is expressed predominantly in the glandular epithelium of cancerous ovaries, but not in normal ovaries of hens. Collectively, these results indicate cell-specific expression of WNT4 in the reproductive tract of chickens and that it likely has crucial roles in development and function of oviduct as well as initiation of ovarian carcinogenesis in laying hens.
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Effects of background fluid on the efficiency of inactivating yeast with non-thermal atmospheric pressure plasma.
PLoS ONE
PUBLISHED: 01-01-2013
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Non-thermal plasma at atmospheric pressure has been actively applied to sterilization. However, its efficiency for inactivating microorganisms often varies depending on microbial species and environments surrounding the microorganisms. We investigated the influence of environmental factors (surrounding media) on the efficiency of microbial inactivation by plasma using an eukaryotic model microbe, Saccharomyces cerevisiae, to elucidate the mechanisms for differential efficiency of sterilization by plasma. Yeast cells treated with plasma in water showed the most severe damage in viability and cell morphology as well as damage to membrane lipids, and genomic DNA. Cells in saline were less damaged compared to those in water, and those in YPD (Yeast extract, Peptone, Dextrose) were least impaired. HOG1 mitogen activated protein kinase was activated in cells exposed to plasma in water and saline. Inactivation of yeast cells in water and saline was due to the acidification of the solutions by plasma, but higher survival of yeast cells treated in saline may have resulted from the additional effect related to salt strength. Levels of hydroxyl radical (OH·) produced by plasma were the highest in water and the lowest in YPD. This may have resulted in differential inactivation of yeast cells in water, saline, and YPD by plasma. Taken together, our data suggest that the surrounding media (environment) can crucially affect the outcomes of yeast cell plasma treatment because plasma modulates vital properties of media, and the toxic nature of plasma can also be altered by the surrounding media.
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Hypermethylation and post-transcriptional regulation of DNA methyltransferases in the ovarian carcinomas of the laying hen.
PLoS ONE
PUBLISHED: 01-01-2013
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DNA methyltransferases (DNMTs) are key regulators of DNA methylation and have crucial roles in carcinogenesis, embryogenesis and epigenetic modification. In general, DNMT1 has enzymatic activity affecting maintenance of DNA methylation, whereas DNMT3A and DNMT3B are involved in de novo methylation events. Although DNMT genes are well known in mammals including humans and mice, they are not well studied in avian species, especially the laying hen which is recognized as an excellent animal model for research on human ovarian carcinogenesis. Results of the present study demonstrated that expression of DNMT1, DNMT3A and DNMT3B genes was significantly increased, particularly in the glandular epithelia (GE) of cancerous ovaries, but not normal ovaries. Consistent with this result, immunoreactive 5-methylcytosine protein was predominantly abundant in nuclei of stromal and GE cells of cancerous ovaries, but it was also found that, to a lesser extent, in nuclei of stromal cells of normal ovaries. Methylation-specific PCR analysis detected hypermethylation of the promoter regions of the tumor suppressor genes in the initiation and development of chicken ovarian cancer. Further, several microRNAs, specifically miR-1741, miR-16c, and miR-222, and miR-1632 were discovered to influence expression of DNMT3A and DNMT3B, respectively, via their 3-UTR which suggests post-transcriptional regulation of their expression in laying hens. Collectively, results of the present study demonstrated increased expression of DNMT genes in cancerous ovaries of laying hens and post-transcriptional regulation of those genes by specific microRNAs, as well as control of hypermethylation of the promoters of tumor suppressor genes.
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Application of an enzyme-based biofuel cell containing a bioelectrode modified with deoxyribonucleic acid-wrapped single-walled carbon nanotubes to serum.
Enzyme Microb. Technol.
PUBLISHED: 11-25-2011
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Enzyme-based biofuel cells (EFCs) are a form of biofuel cells (BFCs) that can utilize redox enzymes as biocatalysts. Applications of an EFC to an implantable system are evaluated under mild conditions, such as ambient temperature or neutral pH. In the present study, an EFC containing a bioelectrode modified with deoxyribonucleic acid (DNA)-wrapped single-walled carbon nanotubes (SWNTs) was applied to a serum system. The protection of immobilized glucose oxidase (GOD) using DNA-wrapped SWNTs was investigated in a trypsin environment, which can exist in a serum. GOD is immobilized by masking the active site onto the anode electrode. The anode/cathode system in the cell was composed of GOD/laccase as the biocatalysts and glucose/oxygen as the substrates in serum. The electrical properties of the anode in serum according to cyclic voltammetry (CV cycle) were improved using the DNA-wrapped SWNTs. Overall, an EFC that employed DNA-wrapped SWNTs and GOD immobilization in conjunction with protection of the active site increased the stability of GOD in serum, which enabled a high level of power production (ca. 190 ?W/cm(2)) for up to 1 week.
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Dual role of methionyl-tRNA synthetase in the regulation of translation and tumor suppressor activity of aminoacyl-tRNA synthetase-interacting multifunctional protein-3.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 11-21-2011
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Mammalian methionyl-tRNA synthetase (MRS) plays an essential role in initiating translation by transferring Met to initiator tRNA (tRNA(i)(Met)). MRS also provides a cytosolic anchoring site for aminoacyl-tRNA synthetase-interacting multifunctional protein-3 (AIMP3)/p18, a potent tumor suppressor that is translocated to the nucleus for DNA repair upon DNA damage. However, the mechanism by which this enzyme mediates these two seemingly unrelated functions is unknown. Here we demonstrate that AIMP3 is released from MRS by UV irradiation-induced stress. Dissociation was induced by phosphorylation of MRS at Ser662 by general control nonrepressed-2 (GCN2) following UV irradiation. Substitution of Ser662 to Asp (S662D) induced a conformational change in MRS and significantly reduced its interaction with AIMP3. This mutant possessed significantly reduced MRS catalytic activity because of loss of tRNA(Met) binding, resulting in down-regulation of global translation. According to the Met incorporation assay using stable HeLa cells expressing MRS S662A or eukaryotic initiation factor-2 subunit-? (eIF2?) S51A, inactivation of GCN2-induced phosphorylation at eIF2? or MRS augmented the role of the other, suggesting a cross-talk between MRS and eIF2? for efficient translational inhibition. This work reveals a unique mode of regulation of global translation as mediated by aminoacyl-tRNA synthetase, specifically MRS, which we herein identified as a previously unidentified GCN2 substrate. In addition, our research suggests a dual role for MRS: (i) as a coregulator with eIF2? for GCN2-mediated translational inhibition; and (ii) as a coupler of translational inhibition and DNA repair following DNA damage by releasing bound tumor suppressor AIMP3 for its nuclear translocation.
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A case of hypereosinophilic syndrome presenting with multiorgan infarctions associated with disseminated intravascular coagulation.
Allergy Asthma Immunol Res
PUBLISHED: 09-28-2011
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Thromboembolism is one of the most critical complications of hypereosinophilic syndrome (HES). We report here a case of multi-organ infarctions related to HES. A 23-year-old woman was referred to our hospital with hemoptysis. Not only pulmonary, but also renal and splenic infarctions were detected on computed tomography images. Blood tests showed profound peripheral eosinophilia. She was diagnosed with HES with disseminated intravascular coagulation (DIC). We initiated infusion of corticosteroids, which effectively suppressed peripheral eosinophilia. However, consumptive coagulopathy did not improve and intracerebral hemorrhage related to thrombosis then developed. Addition of interferon-alpha resulted in the correction of the DIC associated with HES.
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Chemical modulators working at pharmacological interface of target proteins.
Bioorg. Med. Chem.
PUBLISHED: 09-23-2011
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For last few decades, the active site cleft and substrate-binding site of enzymes as well as ligand-binding site of the receptors have served as the main pharmacological space for drug discovery. However, rapid accumulation of proteome and protein network analysis data has opened a new therapeutic space that is the interface between the interacting proteins. Due to the complexity of the interaction modes and the numbers of the participating components, it is still challenging to identify the chemicals that can accurately control the protein-protein interactions at desire. Nonetheless, the number of chemical drugs and candidates working at the interface of the interacting proteins are rapidly increasing. This review addresses the current case studies and state-of-the-arts in the development of small chemical modulators controlling the interactions of the proteins that have pathological implications in various human diseases such as cancer, immune disorders, neurodegenerative and infectious diseases.
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Clinical progress of epilepsy in children with tuberous sclerosis: prognostic factors for seizure outcome.
Chonnam Med J
PUBLISHED: 07-28-2011
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The incidence and outcome of epilepsy in tuberous sclerosis (TS) patients have not yet been thoroughly investigated. The aim of this study was to evaluate the clinical features and prognosis of epileptic seizures associated with TS. The medical records of 29 patients who satisfied the diagnostic criteria for TS and were followed up for at least 2 years at the Department of Pediatrics, Chonnam National University Hospital (CNUH), between January 2000 and December 2010 were reviewed. Onset age of seizure, initial electroencephalography (EEG) findings, and efficacy of treatment were evaluated. Brain imaging studies were reanalyzed to determine the number of cortical tubers and subependymal nodules present. A total of 26 (89.6%) cases presented with seizures. In the seizure-controlled group (n=9, 34.6%), the mean number of cortical tubers was 4.5 (range, 0-16) and the mean number of subependymal nodules was 6.2 (range, 0-14). Initial EEG identified epileptiform discharges in 4 (44.5%) of these cases. In the seizure-sustained group (n=17, 58.6%), 10 patients had initial seizures before 1 year of age. In this group, the mean number of cortical tubers was 6.0 (range, 0-20) and the mean number of subependymal nodules was 6.0 (range, 1-11). A total of 15 (88.2%) had epileptiform discharges on their initial EEGs. In three patients who did not show any seizures during the observation period, the mean number of cortical tubers was 1.3 (range, 0-2), and the mean number of subependymal nodules was 4.6 (range, 0-13). Medical intractability of epilepsy in conjunction with TS did not correlate with age at onset of seizure, the number of cortical tubers, or subependymal nodules, but was associated with initial EEG findings.
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Hog millet (Panicum miliaceum L.)-supplemented diet ameliorates hyperlipidemia and hepatic lipid accumulation in C57BL/6J-ob/ob mice.
Nutr Res Pract
PUBLISHED: 07-20-2011
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Dietary intake of whole grains reduces the incidence of chronic diseases such as obesity, diabetes, cardiovascular disease, and cancer. In an earlier study, we showed that Panicum miliaceum L. extract (PME) exhibited the highest anti-lipogenic activity in 3T3-L1 cells among extracts of nine different cereal grains tested. In this study, we hypothesized that PME in the diet would lead to weight loss and augmentation of hyperlipidemia by regulating fatty acid metabolism. PME was fed to ob/ob mice at 0%, 0.5%, or 1% (w/w) for 4 weeks. After the experimental period, body weight changes, blood serum and lipid profiles, hepatic fatty acid metabolism-related gene expression, and white adipose tissue (WAT) fatty acid composition were determined. We found that the 1% PME diet, but not the 0.5%, effectively decreased body weight, liver weight, and blood triglyceride and total cholesterol levels (P < 0.05) compared to obese ob/ob mice on a normal diet. Hepatic lipogenic-related gene (PPAR?, L-FABP, FAS, and SCD1) expression decreased, whereas lipolysis-related gene (CPT1) expression increased in animals fed the 1% PME diet (P < 0.05). Long chain fatty acid content and the ratio of C18:1/C18:0 fatty acids decreased significantly in adipose tissue of animals fed the 1% PME diet (P < 0.05). Serum inflammatory mediators also decreased significantly in animals fed the 1% PME diet compared to those of the ob/ob control group (P < 0.05). These results suggest that PME is useful in the chemoprevention or treatment of obesity and obesity-related disorders.
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Differential expression of alpha 2 macroglobulin in response to dietylstilbestrol and in ovarian carcinomas in chickens.
Reprod. Biol. Endocrinol.
PUBLISHED: 07-12-2011
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Alpha 2 macroglobulin (A2M; also known as ovostatin), a homotetrameric protein with four disulfide-linked subunits, has the unique feature of inactivating/inhibiting most known proteases including serine-, threonine-, cysteine-, aspartic- and metalloproteases. In chickens, A2M has been identified and characterized biochemically, but little is known of its functional role(s) in the oviduct, hormonal regulation of expression or its expression in ovarian carcinomas in chickens. Therefore, we investigated estrogen regulation of A2M gene expression during development of the chicken oviduct, and its expression in normal and cancerous ovaries from chickens.
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Stem cell dynamics in an experimental model of stroke.
Chonnam Med J
PUBLISHED: 05-31-2011
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We investigated the migration of endogenous neural stem cells (NSCs) toward an infarct lesion in a photo-thrombotic stroke model. The lesions produced by using rose bengal dye (20 mg/kg) with cold light in the motor cortex of Sprague-Dawley rats were also evaluated with sequential magnetic resonance imaging (MRI) from 30 minutes through 8 weeks. Migration of NSCs was identified by immunohistochemistry for nestin monoclonal antibody in the lesion cortex, subventricular zone (SVZ), and corpus callosum (CC). The contrast to noncontrast ratio (CNR) on MRI was greatest at 12 hours in DWI and decreased over time. By contrast, T1-weighted and T2-weighted images showed a constant CNR from the beginning through 8 weeks. MRI of the lesional cortex correlated with histopathologic findings, which could be divided into three stages: acute (edema and necrosis) within 24 hours, subacute (acute and chronic inflammatory cell infiltration) at 2 to 7 days, and chronic (gliofibrosis) at 2 to 4 weeks. The volume of the infarct was significantly reduced by reparative gliofibrosis. The number of nestin(+) NSCs in the contralateral SVZ was similar to that of the ipsilateral SVZ in each group. However, the number of nestin(+) NSCs in the ipsilateral cortex and CC increased at 12 hours to 3 days compared with the contralateral side (p<0.01) and was reduced significantly by 7 days (p<0.01). Active emigration of internal NSCs from the SVZ toward the infarct lesion may also contribute to decreased volume of the infarct lesion, but the self-repair mechanism by endogenous NSCs is insufficient to treat stroke causing extensive neuronal death. Further studies should be focused on amplification technologies of NSCs to enhance the collection of endogenous or transplanted NSCs for the treatment of stroke.
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Effects of Panicum miliaceum L. extract on adipogenic transcription factors and fatty acid accumulation in 3T3-L1 adipocytes.
Nutr Res Pract
PUBLISHED: 05-18-2011
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The dietary intake of whole grains is known to reduce the incidence of chronic diseases such as obesity, diabetes, cardiovascular disease, and cancer. To investigate whether there are anti-adipogenic activities in various Korean cereals, we assessed water extracts of nine cereals. The results showed that treatment of 3T3-L1 adipocytes with Sorghum bicolor L. Moench, Setaria italica Beauvois, or Panicum miliaceum L. extract significantly inhibited adipocyte differentiation, as determined by measuring oil red-O staining, triglyceride accumulation, and glycerol 3-phosphate dehydrogenase activity. Among the nine cereals, P. miliaceum L. showed the highest anti-adipogenic activity. The effects of P. miliaceum L. on mRNA expression of peroxisome proliferator-activated receptor-?, sterol regulatory element-binding protein 1, and the CCAAT/enhancer binding protein-? were evaluated, revealing that the extract significantly decreased the expression of these genes in a dose-dependent manner. Moreover, P. miliaceum L. extract changed the ratio of monounsaturated fatty acids to saturated fatty acids in adipocytes, which is related to biological activity and cell characteristics. These results suggest that some cereals efficiently suppress adipogenesis in 3T3-L1 adipocytes. In particular, the effect of P. miliaceum L. on adipocyte differentiation is associated with the downregulation of adipogenic genes and fatty acid accumulation in adipocytes.
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Progression detection capability of macular thickness in advanced glaucomatous eyes.
Ophthalmology
PUBLISHED: 04-29-2011
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To evaluate the progression rate of macular and circumpapillary retinal nerve fiber layer (RNFL) thickness in advanced glaucomatous eyes using spectral domain optical coherence tomography (Cirrus HD-OCT, Carl Zeiss Meditec, Dublin, CA).
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