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Find video protocols related to scientific articles indexed in Pubmed.
Formononetin Attenuates Osteoclastogenesis via Suppressing the RANKL-Induced Activation of NF-?B, c-Fos, and Nuclear Factor of Activated T-Cells Cytoplasmic 1 Signaling Pathway.
J. Nat. Prod.
PUBLISHED: 11-15-2014
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Formononetin (1), a plant-derived phytoestrogen, possesses bone protective properties. To address the potential therapeutic efficacy and mechanism of action of 1, we investigated its antiosteoclastogenic activity and its effect on nuclear factor-kappaB ligand (RANKL)-induced bone-marrow-derived macrophages (BMMs). Compound 1 markedly inhibited RANKL-induced osteoclast differentiation in the absence of cytotoxicity, by regulating the expression of osteoprotegerin (OPG) and RANKL in BMMs and in cocultured osteoblasts. Compound 1 significantly inhibited RANKL-induced tumor necrosis factor (TNF)-?, interleukin (IL)-1?, IL-6, monocyte chemoattractant protein-1 (MCP-1), regulated on activation normal T cell expressed and secreted (RANTES), and macrophage inflammatory protein-1? (MIP-1?) in a concentration-dependent manner. These effects were accompanied by a decrease in RANKL-induced activation of the NF-?B p65 subunit, degradation of inhibitor ?B? (I?B?), induction of NF-?B, and phosphorylation of AKT, extracellular-signal regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (p38 MAPK). NF-?B siRNA suppressed AKT, ERK, JNK, and p38 MAPK phosphorylation. Furthermore, 1 significantly suppressed c-Fos and nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), key transcription factors during osteoclastogenesis. SP600125, a specific inhibitor of JNK, reduced RANKL-induced expression of phospho-c-Jun, c-Fos, and NFATc1 and inhibited osteoclast formation. These results suggested that 1 acted as an antiresorption agent by blocking osteoclast activation.
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Red Ginseng Extract Promotes the Hair Growth in Cultured Human Hair Follicles.
J Med Food
PUBLISHED: 11-15-2014
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Abstract Ginseng has been shown to promote hair growth in several recent studies. However, its effects on human hair follicles and its mechanisms of action have not been sufficiently elucidated. This study aimed to investigate the hair growth-promoting effects of red ginseng extract (RGE) and its ginsenosides. The proliferative activities of cultured human hair follicles treated with RGE and ginsenoside-Rb1 were assessed using Ki-67 immunostaining. Their effects on isolated human dermal papilla cells (hDPCs) were evaluated using cytotoxicity assays, immunoblot analysis of signaling proteins, and the determination of associated growth factors. We examined the ability of RGE and ginsenosides to protect hair matrix keratinocyte proliferation against dihydrotestosterone (DHT)-induced suppression and their effects on the expression of androgen receptor. The in vivo hair growth-promoting effect of RGE was also investigated in C57BL/6 mice. Both RGE and ginsenoside-Rb1 enhanced the proliferation of hair matrix keratinocytes. hDPCs treated with RGE or ginsenoside-Rb1 exhibited substantial cell proliferation and the associated phosphorylation of ERK and AKT. Moreover, RGE, ginsenoside-Rb1, and ginsenoside-Rg3 abrogated the DHT-induced suppression of hair matrix keratinocyte proliferation and the DHT-induced upregulation of the mRNA expression of androgen receptor in hDPCs. Murine experiments revealed that the subcutaneous injection of 3% RGE resulted in more rapid hair growth than the negative control. In conclusion, RGE and its ginsenosides may enhance hDPC proliferation, activate ERK and AKT signaling pathways in hDPCs, upregulate hair matrix keratinocyte proliferation, and inhibit the DHT-induced androgen receptor transcription. These results suggest that red ginseng may promote hair growth in humans.
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Correlation between Femoral Guidewire Position and Tunnel Communication in Double Bundle Anterior Cruciate Ligament Reconstruction.
Yonsei Med. J.
PUBLISHED: 10-18-2014
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The object of this study was to determine the shortest possible distances of antero-medial (AM) and postero-lateral (PL) guide wire tunnel positions required to prevent femoral bone tunnel communication in double-bundle anterior cruciate ligament (ACL) reconstruction using human cadaver knees.
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Effect of Anteromedial Portal Entrance Drilling Angle during Anterior Cruciate Ligament Reconstruction: A Three-Dimensional Computer Simulation.
Yonsei Med. J.
PUBLISHED: 10-18-2014
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The object of this study was to evaluate entrance angle effects on femoral tunnel length and cartilage damage during anteromedial portal drilling using three-dimensional computer simulation.
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Highly sensitive immunoassay for the diagnosis of acute myocardial infarction using silica spheres encapsulating a quantum dot layer.
Anal. Chem.
PUBLISHED: 10-07-2014
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Commercial ELISA kits for substance P (SubP), which are helpful for the clinical diagnosis of acute myocardial infarction, are limited in efficacy because of low sensitivity. A highly sensitive immunoassay was developed using silica spheres encapsulating a quantum dot-layer (SQS) and labeling antibodies, on a Parylene A-modified plate. The high sensitivity was possible by taking advantage of the enhanced photoluminescence of the SQS and dense immobilization of SubP on a Parylene A-modified plate. Glutaraldehyde was used for cross-linking of SQS to the anti-SubP antibody and SubP to the Parylene A coating. The SQS-linked immunosorbent assay (SQSLISA) was optimized and validated. The dynamic range for the assay was 1-10000 pg/mL with a linear correlation factor of 0.9992 when the competitive SQSLISA was employed. The intra- and interday accuracies were 93-100% and 87-122%, respectively. The reproducibility was lower than 11%. The developed method was applied to clinical samples collected from healthy controls (n = 30) and acute myocardial infarction (n = 16) and it displayed a high correlation with the commercial ELISA kit, with a limit of detection that was 30-fold lower. Clinical sample analysis confirmed that SubP is a promising diagnostic marker for acute myocardial infarction. The SQSLISA is expected to be a practical and useful assay tool.
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Analysis of an ECG Record Database Reveals QT Interval Prolongation Potential of Famotidine in a Large Korean Population.
Cardiovasc. Toxicol.
PUBLISHED: 09-26-2014
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Some non-antiarrhythmic drugs have the undesirable property of delaying cardiac repolarization, an effect that can be measured empirically as a prolongation of the QT interval by surface electrocardiogram (ECG). The QT prolongation and proarrhythmia potential of famotidine are largely unknown, particularly in individuals that have cardiovascular risk factors such as abnormal electrolyte levels. Based on an analysis of QT/QTc intervals from a database of ECG recordings from a large Korean population (ECG-ViEW, 710,369 ECG recordings from 371,401 individuals), we observed that famotidine administration induced a prolonged QTc interval (above 480 ms, p < 0.05 compared to before-treatment, based on a McNemar test). Furthermore, famotidine induced QT prolongations in 10 out of 14 patients with hypocalcemia and 11 out of 13 patients with hypomagnesemia [difference of mean between before and after famotidine administration; 38.00 ms (95 % confidence interval 2.72-73.28) and 67.08 ms (95 % confidence interval 24.94-109.21), p < 0.05 and p < 0.01 by paired t test, respectively]. In vitro, the IC50 of famotidine for human-ether-a-go-go gene (hERG) channel inhibition was higher than 100 ?M as determined by automated patch clamp hERG current assay, implying that hERG channel inhibition is not the underlying mechanism for QT prolongation. These results suggest that famotidine administration increases a proarrhythmic potential, especially in subjects with electrolytes imbalance.
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Helicobacter pylori Induces Vascular Endothelial Growth Factor Production in Gastric Epithelial Cells Through Hypoxia-Inducible Factor-1?-Dependent Pathway.
Helicobacter
PUBLISHED: 09-18-2014
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Although Helicobacter pylori have been known to induce vascular endothelial growth factor (VEGF) production in gastric epithelial cells, the precise mechanism for cellular signaling is incompletely understood. In this study, we investigated the role of bacterial virulence factor and host cellular signaling in VEGF production of H. pylori-infected gastric epithelial cells.
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Rebuttal to "Life-threatening cardiac tamponade: a rare complication of acupuncture": who framed acupuncture?
J Cardiothorac Surg
PUBLISHED: 08-28-2014
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A rebuttal to Chun KJ, Lee SG, Son BS, Kim Do H: Life-threatening cardiac tamponade: a rare complication of acupuncture. J Cardiothorac Surg 2014, 9:61.
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Zhongshania aliphaticivorans sp. nov., an aliphatic hydrocarbon-degrading bacterium isolated from marine sediment, and transfer of Spongiibacter borealis Jang et al. 2011 to the genus Zhongshania as Zhongshania borealis comb. nov.
Int. J. Syst. Evol. Microbiol.
PUBLISHED: 08-13-2014
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A Gram-staining-negative, facultatively aerobic bacterium, designated SM-2(T), was isolated from a sea-tidal flat of Yellow Sea, South Korea. Cells were catalase- and oxidase-positive motile rods with a single polar flagellum. Growth of strain SM-2(T) was observed at 10-37 °C (optimum, 25-30 °C), at pH 5.5-8.5 (optimum, pH 7.0-7.5) and in the presence of 0-11?% (w/v) NaCl (optimum, 2?%). Strain SM-2(T) contained ubiquinone-8 (Q-8) as the sole isoprenoid quinone and C17?:?1?8c, summed feature 3 (comprising C16?:?1?7c and/or iso-C15?:?0 2-OH), C17?:?0 and C18?:?1?7c as the major fatty acids. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unidentified lipid were identified as the major cellular polar lipids. The G+C content of the genomic DNA was 52.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SM-2(T) formed a tight phyletic lineage with Zhongshania antarctica ZS5-23(T), Zhongshania guokunii ZS6-22(T) and Spongiibacter borealis CL-AS9(T), but that S. borealis CL-AS9(T) was distinct from other species of the genus Spongiibacter. Based on 16S rRNA gene sequence similarities, strain SM-2(T) was most closely related to S. borealis CL-AS9(T), Z. antarctica ZS5-23(T) and Z. guokunii ZS6-22(T), with similarities of 99.5?%, 98.9?% and 98.7?%, respectively, but the DNA-DNA hybridization values among these species were clearly lower than 70?%. On the basis of chemotaxonomic data and molecular properties, we propose strain SM-2(T) represents a novel species of the genus Zhongshania with the name Zhongshania aliphaticivorans sp. nov. (type strain SM-2(T)?=?KACC 18120(T)?=?JCM 30138(T)). We also propose the transfer of Spongiibacter borealis Jang et al. 2011 to the genus Zhongshania as Zhongshania borealis comb. nov. (type strain CL-AS9(T)?=?KCCM 90094(T)?=?JCM 17304(T)).
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Assessment of the quality of reporting in randomised controlled trials of acupuncture in the Korean literature using the CONSORT statement and STRICTA guidelines.
BMJ Open
PUBLISHED: 08-01-2014
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This study aims to assess the completeness of reporting of randomised controlled trials (RCTs) of acupuncture in the Korean literature.
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Susceptibility of Escherichia coli from community-acquired urinary tract infection to fosfomycin, nitrofurantoin, and temocillin in Korea.
J. Korean Med. Sci.
PUBLISHED: 07-30-2014
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With increase of multi-drug resistant Escherichia coli in community-acquired urinary tract infections (CA-UTI), other treatment option with a therapeutic efficacy and a low antibiotic selective pressure is necessary. In this study, we evaluated in vitro susceptibility of E. coli isolates from CA-UTI to fosfomycin (FM), nitrofurantoin (NI), temocillin (TMO) as well as trimethoprim-sulfamethoxazole (SMX), ciprofloxacin (CIP) and cefepime (FEP). The minimal inhibitory concentrations were determined by E-test or agar dilution method according to the Clinical and Laboratory Standards Institute guidelines, using 346 E. coli collected in 12 Korean hospitals from March 2010 to February 2011. FM, NI and TMO showed an excellent susceptibility profile; FM 100% (346/346), TMO 96.8% (335/346), and NI 99.4% (344/346). Conversely, resistance rates of CIP and SMX were 22% (76/346) and 29.2% (101/349), respectively. FEP still retained an activity of 98.5%. In Korea, NI and TMO in addition to FM are a good therapeutic option for uncomplicated CA-UTI, especially for lower UTI.
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Estrogenic Endocrine-Disrupting Chemicals Modulate the Production of Inflammatory Mediators and Cell Viability of Lipopolysaccharide-Stimulated Macrophages.
Inflammation
PUBLISHED: 07-26-2014
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Estrogenic endocrine-disrupting chemicals (EDCs) are exogenous substances that act as competitive inhibitors of estrogen in the endocrine system. By disrupting the endocrine system, EDCs can cause severe disabilities and diseases, including cancers and altered sexual development. Although the influence of these molecules in the endocrine system is evident, the effects of EDCs on the immune system as well as their cytotoxicity have been poorly examined. Therefore, we selected 21 EDCs that are commonly found in Korean ecosystems, such as surface waters and effluents, and studied their immunologic effects by comparing nitric oxide (NO) production and cytotoxicity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells (RAW cells), a macrophage cell line. Among the EDCs tested, fenitrothion (FTH) inhibited the messenger RNA (mRNA) expression of inducible NO synthase (iNOS), resulting in reduced NO production, while treatment with andostenedione (AD), diethyl phthalate, di-n-butyl phthalate (DBP), estriol, or molinate decreased production of NO in an iNOS-independent fashion. In contrast, benzo(a)pyrene (B(a)P) increased the production of NO in RAW cells. In addition, AD, DBP, or FTH inhibited the mRNA expression of tumor necrosis factor alpha or interleukin-1 beta. Treatment with 17-?-ethynylestradiol, 17-?-estradiol, 4-n-butyl phenol, or alachlor induced apoptosis of RAW cells, while dicyclohexyl phthalate and B(a)P caused cell death in an apoptosis-independent manner. These data suggest that EDCs can influence the immune response to pathogens by modulating the functions of macrophages.
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Correlation between Ultrasonography Findings and Electrodiagnostic Severity in Carpal Tunnel Syndrome: 3D Ultrasonography.
J Clin Neurol
PUBLISHED: 07-20-2014
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To determine the correlation between the cross-sectional area (CSA) of the median nerve measured at the wrist using three-dimensional (3D) ultrasonography (US) and the electrophysiological severity of carpal tunnel syndrome (CTS).
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A capacitive biosensor based on an interdigitated electrode with nanoislands.
Anal. Chim. Acta
PUBLISHED: 07-11-2014
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A capacitive biosensor based on an interdigitated electrode (IDE) with nanoislands was developed for label-free detection of antigen-antibody interactions. To enable sensitive capacitive detection of protein adsorption, the nanoislands were fabricated between finger electrodes of the IDE. The effect of the nanoislands on the sensitive capacitive measurement was estimated using horseradish peroxidase (HRP) as a model protein. Additionally, a parylene-A film was coated on the IDE with nanoislands to improve the efficiency of protein immobilization. By using HRP and hepatitis B virus surface antigen (HBsAg) as model analytes, the effect of the parylene-A film on the capacitive detection of protein adsorption was demonstrated.
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Acupuncture for treating acute ankle sprains in adults.
Cochrane Database Syst Rev
PUBLISHED: 06-24-2014
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An acute ankle sprain is a sudden-onset injury of one or more of the ankle ligaments. It is one of the most common musculoskeletal injuries in the general population as well as in athletes. In some countries, such as China and Korea, acupuncture is frequently used in the treatment of ankle sprains, either as a single treatment or a secondary intervention accompanied by standard medical treatment.
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Motion patterns in acupuncture needle manipulation.
Acupunct Med
PUBLISHED: 06-17-2014
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In clinical practice, acupuncture manipulation is highly individualised for each practitioner. Before we establish a standard for acupuncture manipulation, it is important to understand completely the manifestations of acupuncture manipulation in the actual clinic. To examine motion patterns during acupuncture manipulation, we generated a fitted model of practitioners' motion patterns and evaluated their consistencies in acupuncture manipulation.
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Arginine enhances osteoblastogenesis and inhibits adipogenesis through the regulation of Wnt and NFATc signaling in human mesenchymal stem cells.
Int J Mol Sci
PUBLISHED: 06-16-2014
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Arginine, an ?-amino acid, has been reported to exert beneficial effects that ameliorate health problems and prevent excessive fat deposition. In this study, we investigated whether the activation of cell signaling by arginine can induce osteogenic differentiation and modulate excessive adipogenic differentiation in human mesenchymal stem cells (MSCs). Arginine potently induced the expression of type I?1 collagen, osteocalcin, and ALP in a dose-dependent manner without causing cytotoxicity. Arginine significantly increased the mRNA expression of the osteogenic transcription factors runt-related transcription factor 2 (Runx2), DIx5, and osterix. Furthermore, arginine demonstrated its antiadipogenicity by decreasing adipocyte formation and triglyceride (TG) content in MSCs and inhibiting the mRNA expression of the adipogenic transcription factors peroxisome proliferator-activated receptor ? (PPAR?), CCAAT/enhancer-binding protein ? (C/EBP?), and fatty acid binding protein 4 (Fabp4). This effect was associated with increased expression of Wnt5a, and nuclear factor of activated T-cells (NFATc), and was abrogated by antagonists of Wnt and NFATc, which indicated a role of Wnt and NFATc signaling in the switch from adipogenesis to osteoblastogenesis induced by arginine. In conclusion, this is the first report of the dual action of arginine in promoting osteogenesis and inhibiting adipocyte formation through involving Wnt5a and NFATc signaling pathway.
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The Standardized BHH10 Extract, a Combination of Astragalus membranaceus, Cinnamomum cassia, and Phellodendron amurense, Reverses Bone Mass and Metabolism in a Rat Model of Postmenopausal Osteoporosis.
Phytother Res
PUBLISHED: 05-26-2014
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Jasin-hwan-gagambang (BHH10), a modified prescription of Jasin-hwan, contains Astragalus membranaceus, Cinnamomum cassia, and Phellodendron amurense, and it has been traditionally used to treat osteoporosis and other inflammatory diseases. In this study, we systematically investigated the protective effects of BHH10 in ovariectomy (OVX)-induced rats. Sprague-Dawley rats were randomly divided into sham and OVX subgroups. The rats in the OVX group were treated with vehicle, BHH10, alendronate (ALN), and 17?-estradiol (E2). BHH10 treatment significantly inhibited OVX-induced increases in body weight and uterus atrophy. In addition, it significantly increased the bone mineral density (BMD) and prevented a decrease in trabecular bone volume, connectivity density, trabecular number, thickness, and separation at the total femur and femur neck. The OVX rats showed significant decreases in the serum levels of calcium and phosphorous and significant increases in the serum levels of cholesterol, low-density lipoprotein cholesterol, alkaline phosphatase, osteocalcin, C-telopeptide type 1 collagen, and bone morphogenetic protein-2. These changes were significantly reduced to near sham levels by administration of BHH10 to OVX rats. BHH10-treated rats had a greater bone mass, a better structural architecture of the bone, and higher levels of biochemical markers of the bone than did the ALN-treated or E2-treated rats. These results suggest that BHH10 reverses osteoporosis in OVX rats by stimulating bone formation or regulating bone resorption and is not associated with toxicity. © 2014 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd.
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Ultrasonic isolation of the outer membrane of Escherichia coli with autodisplayed Z-domains.
Enzyme Microb. Technol.
PUBLISHED: 05-19-2014
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The outer membrane of Escherichia coli was previously isolated as a liposome-like outer membrane particle using an enzymatic treatment for lysozymes; for immunoassays, the particles were subsequently layered on solid supports via hydrophobic interactions. This work presents an enzyme-free isolation method for the E. coli outer membrane with autodisplayed Z-domains using ultrasonication. First, the properties of the outer membrane particle, such as the particle size, zeta potential, and total protein, were compared with the properties of particles obtained using the previous preparation methods. Compared with the conventional isolation method using an enzyme treatment, the ultrasonic method exhibited a higher efficiency at isolating the outer membrane and less contamination by cytosolic proteins. The isolated outer membrane particles were layered on a gold surface, and the roughness and thickness of the layered outer membrane layers were subsequently analyzed using AFM analysis. Finally, the antibody-binding activity of two outer membrane layers with autodisplayed Z-domains created from particles that were isolated using the enzymatic and ultrasonic isolation methods was measured using fluorescein-labeled antibody as a model analyte, and the activity of the outer membrane layer that was isolated from the ultrasonic method was estimated to be more than 20% higher than that from the conventional enzymatic method.
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Clinically useful predictors of resistance to intravenous immunoglobulin and prognosis of coronary artery lesions in patients with incomplete kawasaki disease.
Korean Circ J
PUBLISHED: 05-15-2014
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The prevalence of incomplete Kawasaki disease (iKD) is progressively increasing. We aimed to retrospectively investigate the predictors of intravenous immunoglobulin (IVIG) resistance in iKD patients and compare them with those of IVIG resistance in complete Kawasaki disease (cKD) patients. We also compared the prognosis of coronary artery lesions (CALs) between the IVIG non-responders and responders in both iKD and cKD groups.
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Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of small volatile molecules using a parylene-matrix chip.
Rapid Commun. Mass Spectrom.
PUBLISHED: 05-12-2014
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In matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS), volatile small molecules have been nearly impossible to analyze because (1) such molecules evaporate under drying and vacuum conditions and (2) the organic matrix creates matrix peaks in the low mass-to-charge (m/z) range (m/z <500). In this work, the analysis of volatile small molecules using MALDI-TOFMS was realized using (1) a parylene-matrix chip to eliminate the matrix peaks of the organic matrix and (2) graphene for the effective adsorption of the small volatile molecules.
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Distinct roles of ROCK1 and ROCK2 during development of porcine preimplantation embryos.
Reproduction
PUBLISHED: 05-06-2014
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Cell-to-cell contact mediated by cell adhesion is fundamental to the compaction process that ensures blastocyst quality during embryonic development. In this study, we first showed that Rho-associated coiled-coil protein kinases (ROCK1 and ROCK2) were expressed both in porcine oocytes and IVF preimplantation embryos, playing different roles in oocytes maturation and embryo development. The amount of mRNA encoding ROCK1 and the protein concentration clearly increased between the eight-cell and morula stages, but decreased significantly when blastocysts were formed. Conversely, ROCK2 was more abundant in the blastocyst compared with other embryonic stages. Moreover, immunostaining showed that ROCK1 protein distribution changed as the embryo progressed through cleavage and compaction to the morula stage. Initially, the protein was predominantly associated with the plasma membrane but later became cytoplasmic. By contrast, ROCK2 protein was localized in both the cytoplasm and the spindle rotation region during oocyte meiosis, but in the cytoplasm and nucleus as the embryo developed. In addition, ROCK2 was present in the trophectoderm cells of the blastocyst. Treatment with 15??M Y27632, a specific inhibitor of ROCKs, completely blocked further development of early four-cell stage embryos. Moreover, we did not detect the expression of ROCK1 but did detect ROCK2 expression in blastocysts. Moreover, lysophosphatidic acid an activator of ROCKs significantly improved the rates of blastocyst formation. These data demonstrate that ROCKs are required for embryo development to the blastocyst stage. Together, our results indicate that ROCK1 and ROCK2 may exert different biological functions during the regulation of compaction and in ensuring development of porcine preimplantation embryos to the blastocyst stage.
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Nod2 and Rip2 contribute to innate immune responses in mouse neutrophils.
Immunology
PUBLISHED: 04-15-2014
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Nod-like receptors are a family of innate immune receptors that link cytosolic sensing of microbial and danger stimuli to the activation of immune responses. Two Nod-like receptor family members, Nod1 and Nod2, recognize bacterial peptidoglycan and activate immune responses via nuclear factor-?B (NF-?B) and mitogen-activated protein kinase (MAPK). The function of Nod1 and Nod2 has been largely studied in macrophages, but the role of these receptors in other innate immune cells remains unclear. In this study, we examined the function of Nod1 and Nod2 in innate immune responses of neutrophils. Mice were injected intraperitoneally with thioglycollate, and then peritoneal neutrophils were isolated 4 hr after injection. Tri-DAP and muramyl-dipeptide (MDP) were used as Nod1 and Nod2 agonists, respectively. The level of cytokines [interleukin-6 (IL-6) and tumour necrosis factor-? (TNF-?)] and chemokines (CXCL1 and CCL2) was increased by MDP, but not Tri-DAP in wild-type (WT) neutrophils. Increased production of cytokines and chemokines with MDP was abolished in Nod2- and Rip2-deficient neutrophils. MDP also induced the activation of NF-?B and MAPK in WT neutrophils, but not in Nod2- and Rip2-deficient cells. Flow cytometry analysis showed that L-selectin shedding was induced by MDP in WT neutrophils, but not in Nod2- and Rip2-deficient cells. MDP and Toll-like receptor (TLR) agonists (Pam3 CSK4 and lipopolysaccharide) exerted synergistic effects on the production of IL-6 and CXCL1 in neutrophils. Moreover, Nod2 and TLR4 cooperated to produce IL-6, TNF-?, CXCL1 and CCL2 in neutrophils in response to Gram-negative bacteria. Our findings suggest that the Nod2-Rip2 axis may contribute to the innate immune response of neutrophils against bacterial infection.
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Anti-inflammatory effects of betaine on AOM/DSS?induced colon tumorigenesis in ICR male mice.
Int. J. Oncol.
PUBLISHED: 04-14-2014
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Betaine is an important human nutrient obtained from various foods and studies in animals and humans have provided results suggesting their pathogenesis of various chronic diseases and points to a role in risk assessment and disease prevention. However, the molecular mechanisms of its activity remain poorly understood and warrant further investigation. This study was performed to investigate the anti-inflammation and tumor preventing capacity of betaine on colitis-associated cancer in mice. In in vivo experiments, we induced colon tumors in mice by azoxymethane (AOM) and dextran sulfate sodium (DSS) and evaluated the effects of betaine on tumor growth. Administration with betaine significantly decreased the incidence of tumor formation with downregulation of inflammation. Treatment with betaine inhibited ROS generation and GSSG concentration in colonic mucosa. Based on the qPCR data, administration of betaine inhibited inflammatory cytokines such TNF-?, IL-6, iNOS and COX-2. In in vitro experiments, LPS-induced NF-?B and inflammatory-related cytokines were inhibited by betaine treatment in RAW 264.7 murine macrophage cells. Our findings suggest that betaine is one of the candidates for the prevention of inflammation-associated colon carcinogenesis.
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A randomized clinical trial of high-dosage coenzyme Q10 in early Parkinson disease: no evidence of benefit.
, M Flint Beal, David Oakes, Ira Shoulson, Claire Henchcliffe, Wendy R Galpern, Richard Haas, Jorge L Juncos, John G Nutt, Tiffini Smith Voss, Bernard Ravina, Clifford M Shults, Karen Helles, Victoria Snively, Mark F Lew, Brian Griebner, Arthur Watts, Shan Gao, Emmanuelle Pourcher, Louisette Bond, Katie Kompoliti, Pinky Agarwal, Cherissa Sia, Mandar Jog, Linda Cole, Munira Sultana, Roger Kurlan, Irene Richard, Cheryl Deeley, Cheryl H Waters, Angel Figueroa, Ani Arkun, Matthew Brodsky, William G Ondo, Christine B Hunter, Joohi Jimenez-Shahed, Alicia Palao, Janis M Miyasaki, Julie So, James Tetrud, Liza Reys, Katharine Smith, Carlos Singer, Anita Blenke, David S Russell, Candace Cotto, Joseph H Friedman, Margaret Lannon, Lin Zhang, Edward Drasby, Rajeev Kumar, Thyagarajan Subramanian, Donna Stuppy Ford, David A Grimes, Diane Cote, Jennifer Conway, Andrew D Siderowf, Marian Leslie Evatt, Barbara Sommerfeld, Abraham N Lieberman, Michael S Okun, Ramon L Rodriguez, Stacy Merritt, Camille Louise Swartz, W R Wayne Martin, Pamela King, Natividad Stover, Stephanie Guthrie, Ray L Watts, Anwar Ahmed, Hubert H Fernandez, Adrienna Winters, Zoltan Mari, Ted M Dawson, Becky Dunlop, Andrew S Feigin, Barbara Shannon, Melissa Jill Nirenberg, Mattson Ogg, Samuel A Ellias, Cathi-Ann Thomas, Karen Frei, Ivan Bodis-Wollner, Sofya Glazman, Thomas Mayer, Robert A Hauser, Rajesh Pahwa, April Langhammer, Ranjit Ranawaya, Lorelei Derwent, Kapil D Sethi, Buff Farrow, Rajan Prakash, Irene Litvan, Annette Robinson, Alok Sahay, Maureen Gartner, Vanessa K Hinson, Samuel Markind, Melisa Pelikan, Joel S Perlmutter, Johanna Hartlein, Eric Molho, Sharon Evans, Charles H Adler, Amy Duffy, Marlene Lind, Lawrence Elmer, Kathy Davis, Julia Spears, Stephanie Wilson, Maureen A Leehey, Neal Hermanowicz, Shari Niswonger, Holly A Shill, Sanja Obradov, Alex Rajput, Marilyn Cowper, Stephanie Lessig, David Song, Deborah Fontaine, Cindy Zadikoff, Karen Williams, Karen A Blindauer, Jo Bergholte, Clara Schindler Propsom, Mark A Stacy, Joanne Field, Dragos Mihaila, Mark Chilton, Ergun Y Uc, Jeri Sieren, David K Simon, Lauren Kraics, Althea Silver, James T Boyd, Robert W Hamill, Christopher Ingvoldstad, Jennifer Young, Karen Thomas, Sandra K Kostyk, Joanne Wojcieszek, Ronald F Pfeiffer, Michel Panisset, Monica Beland, Stephen G Reich, Michelle Cines, Nancy Zappala, Jean Rivest, Richard Zweig, L Pepper Lumina, Colette Lynn Hilliard, Stephen Grill, Marye Kellermann, Paul Tuite, Susan Rolandelli, Un Jung Kang, Joan Young, Jayaraman Rao, Maureen M Cook, Lawrence Severt, Karyn Boyar.
JAMA Neurol
PUBLISHED: 03-26-2014
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Coenzyme Q10 (CoQ10), an antioxidant that supports mitochondrial function, has been shown in preclinical Parkinson disease (PD) models to reduce the loss of dopamine neurons, and was safe and well tolerated in early-phase human studies. A previous phase II study suggested possible clinical benefit.
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Characterization of a novel otubain-like cysteine protease of Cryptosporidium parvum.
Parasitol. Int.
PUBLISHED: 03-22-2014
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Otubains are a recently discovered family of cysteine proteases that participate in the ubiquitin pathway. Here, we partially characterized the biochemical properties of a cysteine protease of Cryptosporidium parvum, which is closely related to otubains. The gene encoding otubain-like cysteine protease of C. parvum (CpOTU) contained the aspartate, cysteine and histidine residues that form the catalytic triad of otubains. The modified ubiquitin-associated domain and LxxL motif were identified in CpOTU. The recombinant CpOTU showed the isopeptidase activity at neutral pH values and its activity was effectively inhibited by ubiquitin aldehyde, N-ethylmaleimide and iodoacetic acid. Interestingly, CpOTU had an unusual C-terminal extension of 217 amino acids compared to mammalian otubains, and the C-terminal extension is essential for the activity of the enzyme. Expression of CpOTU peaked in the oocyst stage of the parasite, which suggested its potential physiological role for the oocyst stage.
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Molecular analysis of Anisakis type I larvae in marine fish from three different sea areas in Korea.
Korean J. Parasitol.
PUBLISHED: 03-21-2014
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Anisakiasis, a human infection of Anisakis L3 larvae, is one of the common foodborne parasitic diseases in Korea. Studies on the identification of anisakid larvae have been performed in the country, but most of them have been focused on morphological identification of the larvae. In this study, we analyzed the molecular characteristics of 174 Anisakis type I larvae collected from 10 species of fish caught in 3 different sea areas in Korea. PCR-RFLP and sequence analyses of rDNA ITS and mtDNA cox1 revealed that the larvae showed interesting distribution patterns depending on fish species and geographical locations. Anisakis pegreffii was predominant in fish from the Yellow Sea and the South Sea. Meanwhile, both A. pegreffii and A. simplex sensu stricto (A. simplex s.str.) larvae were identified in fish from the East Sea, depending on fish species infected. These results suggested that A. pegreffii was primarily distributed in a diverse species of fish in 3 sea areas around Korea, but A. simplex s.str. was dominantly identified in Oncorhynchus spp. in the East Sea.
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Comparison of Supplemented Brucella Agar and Modified Clostridium difficile Agar for Antimicrobial Susceptibility Testing of Clostridium difficile.
Ann Lab Med
PUBLISHED: 03-03-2014
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Antimicrobial susceptibility testing (AST) of Clostridium difficile is increasingly important because of the rise in resistant strains. The standard medium for the AST of C. difficile is supplemented Brucella agar (sBA), but we found that the growth of C. difficile on sBA was not optimal. Because active growth is critical for reliable AST, we developed a new, modified C. difficile (mCD) agar. C. difficile grew better on mCD agar than on sBA.
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Top-down synthesized TiO2 nanowires as a solid matrix for surface-assisted laser desorption/ionization time-of-flight (SALDI-TOF) mass spectrometry.
Anal. Chim. Acta
PUBLISHED: 02-25-2014
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Top-down synthesized TiO2 nanowires are presented as an ideal solid matrix to analyze small biomolecules at a m/z of less than 500. The TiO2 nanowires were synthesized as arrays using a modified hydrothermal process directly on the surface of a Ti plate. Finally, the feasibility of the TiO2 nanowires in the anatase phase as a solid matrix. The crystal and electronic structures of the top-down TiO2 nanowires were analyzed at each step of the hydrothermal process, and the optimal TiO2 nanowires were identified by checking their performance toward the ionization of analytes in surface-assisted laser desorption/ionization time-of-flight (SALDI-TOF) mass spectrometry. Finally, the feasibility of the TiO2 nanowires in the anatase phase as a solid matrix for SALDI-TOF mass spectrometry was demonstrated using eight types of amino acids and peptides as model analytes.
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Ultrasonographic diaphragmatic motion analysis and its correlation with pulmonary function in hemiplegic stroke patients.
Ann Rehabil Med
PUBLISHED: 02-25-2014
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To evaluate diaphragmatic motion via M-mode ultrasonography and to correlate it with pulmonary function in stroke patients.
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Apigenin-induced apoptosis is enhanced by inhibition of autophagy formation in HCT116 human colon cancer cells.
Int. J. Oncol.
PUBLISHED: 02-24-2014
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Apigenin (4',5,7-trihydroxyflavone) is a natural flavonoid, shown to have chemopreventive and/or anticancer properties in a variety of human cancer cells. The involvement of autophagy in apigenin-induced apoptotic cell death of HCT116 human colon cancer cells was investigated. Apigenin induced suppression of cell growth in a concentration-dependent manner in HCT116 cells. Flow cytometric analyses indicated that apigenin resulted in G2/M phase arrest. This flavone also suppressed the expression of both cyclin B1 and its activating partners, Cdc2 and Cdc25c, whereas the expression of cell cycle inhibitors, such as p53 and p53-dependent p21(CIP1/WAF1), was increased after apigenin treatment. Apigenin induced poly (ADP-ribose) polymerase (PARP) cleavage and decreased the levels of procaspase-8, -9 and -3. In addition, the apigenin-treated cells exhibited autophagy, as characterized by the appearance of autophagosomes under fluorescence microscopy and the accumulation of acidic vesicular organelles by flow cytometry. Furthermore, the results of the western blot analysis revealed that the levels of LC3-II, the processed form of LC3-I, was increased by apigenin. Treatment with the autophagy inhibitor 3-methyladenine (3-MA) significantly enhanced the apoptosis induced by apigenin, which was accompanied by an increase in the levels of PARP cleavage. These results indicate that apigenin has apoptosis- and autophagy-inducing effects in HCT116 colon cancer cells. Autophagy plays a cytoprotective role in apigenin-induced apoptosis, and the combination of apigenin and an autophagy inhibitor may be a promising strategy for colon cancer control.
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Diverse Toll-like receptors mediate cytokine production by Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans in macrophages.
Infect. Immun.
PUBLISHED: 02-24-2014
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Toll-like receptors (TLRs) orchestrate a repertoire of immune responses in macrophages against various pathogens. Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans are two important periodontal pathogens. In the present study, we investigated TLR signaling regulating cytokine production of macrophages in response to F. nucleatum and A. actinomycetemcomitans. TLR2 and TLR4 are redundant in the production of cytokines (interleukin-6 [IL-6] and tumor necrosis factor alpha [TNF-?]) in F. nucleatum- and A. actinomycetemcomitans-infected macrophages. The production of cytokines by macrophages in response to F. nucleatum and A. actinomycetemcomitans infection was impaired in MyD88-deficient macrophages. Moreover, cytokine concentrations were lower in MyD88-deficient macrophages than in TLR2/TLR4 (TLR2/4) double-deficient cells. An endosomal TLR inhibitor, chloroquine, reduced cytokine production in TLR2/4-deficient macrophages in response to F. nucleatum and A. actinomycetemcomitans, and DNA from F. nucleatum or A. actinomycetemcomitans induced IL-6 production in bone marrow-derived macrophages (BMDMs), which was abolished by chloroquine. Western blot analysis revealed that TLR2/4 and MyD88 were required for optimal activation of NF-?B and mitogen-activated protein kinases (MAPKs) in macrophages in response to F. nucleatum and A. actinomycetemcomitans, with different kinetics. An inhibitor assay showed that NF-?B and all MAPKs (p38, extracellular signal-regulated kinase [ERK], and Jun N-terminal protein kinase [JNK]) mediate F. nucleatum-induced production of cytokines in macrophages, whereas NF-?B and p38, but not ERK and JNK, are involved in A. actinomycetemcomitans-mediated cytokine production. These findings suggest that multiple TLRs may participate in the cytokine production of macrophages against periodontal bacteria.
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Striatal cholinergic cell ablation attenuates L-DOPA induced dyskinesia in Parkinsonian mice.
J. Neurosci.
PUBLISHED: 02-21-2014
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3,4-Dihydroxyphenyl-L-alanine (L-DOPA)-induced dyskinesia (LID) is a debilitating side effect of long-term dopamine replacement therapy in Parkinson's Disease. At present, there are few therapeutic options for treatment of LID and mechanisms contributing to the development and maintenance of these drug-induced motor complications are not well understood. We have previously shown that pharmacological reduction of cholinergic tone attenuates the expression of LID in parkinsonian mice with established dyskinesia after chronic L-DOPA treatment. The present study was undertaken to provide anatomically specific evidence for the role of striatal cholinergic interneurons by ablating them before initiation of L-DOPA treatment and determining whether it decreases LID. We used a novel approach to ablate striatal cholinergic interneurons (ChIs) via Cre-dependent viral expression of the diphtheria toxin A subunit (DT-A) in hemiparkinsonian transgenic mice expressing Cre recombinase under control of the choline acetyltransferase promoter. We show that Cre recombinase-mediated DT-A ablation selectively eliminated ChIs when injected into striatum. The depletion of ChIs markedly attenuated LID without compromising the therapeutic efficacy of L-DOPA. These results provide evidence that ChIs play a key and selective role in LID and that strategies to reduce striatal cholinergic tone may represent a promising approach to decreasing L-DOPA-induced motor complications in Parkinson's disease.
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Loss of PINK1 attenuates HIF-1? induction by preventing 4E-BP1-dependent switch in protein translation under hypoxia.
J. Neurosci.
PUBLISHED: 02-21-2014
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Parkinson's disease (PD) has multiple proposed etiologies with implication of abnormalities in cellular homeostasis ranging from proteostasis to mitochondrial dynamics to energy metabolism. PINK1 mutations are associated with familial PD and here we discover a novel PINK1 mechanism in cellular stress response. Using hypoxia as a physiological trigger of oxidative stress and disruption in energy metabolism, we demonstrate that PINK1(-/-) mouse cells exhibited significantly reduced induction of HIF-1? protein, HIF-1? transcriptional activity, and hypoxia-responsive gene upregulation. Loss of PINK1 impairs both hypoxia-induced 4E-BP1 dephosphorylation and increase in the ratio of internal ribosomal entry site (IRES)-dependent to cap-dependent translation. These data suggest that PINK1 mediates adaptive responses by activating IRES-dependent translation, and the impairments in translation and the HIF-1? pathway may contribute to PINK1-associated PD pathogenesis that manifests under cellular stress.
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Usefulness of videofluoroscopic swallow study in treacher collins syndrome with cleft palate: a case report.
Ann Rehabil Med
PUBLISHED: 02-05-2014
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A 3-year-old girl had multiple anomalies compatible with Treacher Collins Syndrome (TCS). From the neonatal period, sucking was poor, making tube feeding necessary. Excessive saliva was retained in the oral cavity. Nasal leakage caused by the cleft palate was observed when she spoke. The initial videofluoroscopic swallow study (VFSS) showed a poor posterior bolus transit and nasopharyngeal regurgitation. A delayed swallow reflex and bolus stasis at the vallecular and pyriform sinuses were recognized. Based on the VFSS findings, the patient underwent palatoplasty at 20 months of age. At approximately 23 months of age, a follow-up VFSS was performed; poor posterior bolus transit, nasopharyngeal regurgitation, and delayed swallow reflex were not observed. Finally, the patient was able to eat ground or chopped foods and solid foods orally. We deem VFSS to be helpful in deciding the appropriate management of dysphagia in TCS.
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Toll?like receptor 2 promotes bacterial clearance during the initial stage of pulmonary infection with Acinetobacter baumannii.
Mol Med Rep
PUBLISHED: 02-05-2014
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Toll?like receptor 2 (TLR2) has been identified as a sensor for bacterial lipoprotein. To determine the role of TLR2 in host defense against Acinetobacter baumannii infection, wild?type (WT) and TLR2?deficient mice were infected intranasally with A. baumannii. Body weight, cytokine and chemokine levels in bronchoalveolar (BAL) fluid and lung histopathology were examined. Body weight changes in TLR2?deficient mice were comparable to those of WT mice throughout the experimental period. However, TLR2?deficient mice exhibited an increased bacterial load in the lungs and increased levels of interleukin?6 and chemokine (C?X?C motif) ligand 2 in BAL fluids compared with WT mice 1 day after infection. Histopathological features of lung tissue in WT and TLR2?deficient mice were comparable between WT and TLR2?deficient mice. Results of the present study demonstrate that TLR2 may have a minimal role in the host defense against A. baumannii at the early stages of infection.
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Ameliorative effect of myricetin on insulin resistance in mice fed a high-fat, high-sucrose diet.
Nutr Res Pract
PUBLISHED: 02-04-2014
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Obesity-associated insulin resistance is a strong risk factor for type 2 diabetes mellitus. The aim of this study was to investigate the effect of myricetin on adiposity, insulin resistance, and inflammatory markers in mice with diet-induced insulin resistance.
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The effectiveness and safety of moxibustion for treating cancer-related fatigue: a systematic review and meta-analyses.
Support Care Cancer
PUBLISHED: 02-03-2014
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Among cancer patients, cancer-related fatigue (CRF) is one of the most common symptoms and adversely affects physical ability and quality of life even several years after treatment. This study aims to evaluate the current evidence for moxibustion in patients with CRF.
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Corosolic acid induces apoptotic cell death in HCT116 human colon cancer cells through a caspase-dependent pathway.
Int. J. Mol. Med.
PUBLISHED: 01-21-2014
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Corosolic acid (CA), a pentacyclic triterpene isolated from Lagerstroemia speciosa L. (also known as Banaba), has been shown to exhibit anticancer properties in various cancer cell lines. However, the anticancer activity of CA on human colorectal cancer cells and the underlying mechanisms remain to be elucidated. In this study, we investigated the effects of CA on cell viability and apoptosis in HCT116 human colon cancer cells. CA dose-dependently inhibited the viability of HCT116 cells. The typical hallmarks of apoptosis, such as chromatin condensation, a sub-G1 peak and phosphatidylserine externalization were detected by Hoechst 33342 staining, flow cytometry and Annexin V staining following treatment with CA. Western blot analysis revealed that CA induced a decrease in the levels of procaspase-8, -9 and -3 and the cleavage of poly(ADP-ribose) polymerase (PARP). The apoptotic cell death induced by CA was accompanied by the activation of caspase-8, -9 and -3, which was completely abrogated by the pan-caspase inhibitor, z-VAD?FMK. Furthermore, CA upregulated the levels of pro-apoptotic proteins, such as Bax, Fas and FasL and downregulated the levels of anti-apoptotic proteins, such as Bcl-2 and survivin. Taken together, our data provide insight into the molecular mechanisms of CA-induced apoptosis in colorectal cancer (CRC), rendering this compound a potential anticancer agent for the treatment of CRC.
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Assessment of the quality of reporting for treatment components in Cochrane reviews of acupuncture.
BMJ Open
PUBLISHED: 01-21-2014
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High-quality reporting of treatment details can aid replication of study results in real-world clinical practice. The Standards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA) is a reporting guideline for key elements of acupuncture interventions in clinical trials. This study used STRICTA to investigate whether Cochrane reviews of acupuncture adequately report important treatment details.
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Enterostomy closure timing for minimizing postoperative complications in premature infants.
Pediatr Neonatol
PUBLISHED: 01-18-2014
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For premature infants with advanced acute abdomen, creating a temporary enterostomy is believed to be an appropriate surgical management. However, there is no consensus regarding the timing of enterostomy reversal. The aim of this study was to determine the optimal timing for enterostomy closure (EC) by analyzing EC-related complications.
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MHY-449, a novel dihydrobenzofuro[4,5-b][1,8] naphthyridin-6-one derivative, induces apoptotic cell death through modulation of Akt/FoxO1 and ERK signaling in PC3 human prostate cancer cells.
Int. J. Oncol.
PUBLISHED: 01-10-2014
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Previously, we reported on the anticancer effect of the diastereoisomeric compound MHY-449, a novel dihydro-benzofuro[4,5-b][1,8] naphthyridin-6-one derivative, in HCT116 human colon cancer cells. In the current study, we investigated whether MHY-449 has anticancer effect in prostate cancer cells, and if so, what the molecular mechanisms are. We examined the growth inhibitory effect of MHY-449 on p53 wild?type (p53-wt) LNCaP (androgen?dependent) and p53-null PC3 (androgen-independent) prostate cancer cells. MHY-449 treatment in androgen-independent and p53-null PC3 cells resulted in inhibition of cell growth and induction of apoptosis in a concentration-dependent manner. However, MHY-449 did not show any significant effects on the growth inhibition and apoptotic cell death in androgen-dependent and p53-wt LNCaP cells. Therefore, we used PC3 cells for further studies. The induction of apoptosis in PC3 cells was observed by decreased viability, DNA fragmentation, cleavage of poly (ADP-ribose) polymerase, activations of caspase-3, -8 and -9, and alteration in the ratio of Bax/Bcl-2 protein expression. In addition, MHY-449 induced increase of late apoptosis and sub-G1 DNA which were observed by flow cytometry analysis. Furthermore, MHY-449 reduced the phosphorylation of Akt and FoxO1 and induced the translocation of FoxO1 from cytoplasm to nucleus as shown by western blot analysis. MHY-449 treatment activated extracellular signal-regulated kinase (ERK) signaling in a concentration-dependent manner. MHY-449-induced apoptosis was partially prevented by pretreatment with the ERK inhibitor PD98059 suggesting involvement of ERK in the MHY-449-induced apoptosis. Taken together, these findings suggest that MHY-449 induces apoptosis via downregulation of the Akt/FoxO1 and activation of ERK in androgen-independent, p53-null and PTEN-negative PC3 human prostate cancer cells.
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Performance characteristic of anti-cyclic citrullinated peptide (CCP) assay on Korean rheumatoid arthritis (RA) patients and healthy controls.
J Pharm Biomed Anal
PUBLISHED: 01-06-2014
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The objective of this study was to evaluate the performance of the established anti-cyclic citrullinated peptide (anti-CCP) enzyme-linked immunosorbent assay (ELISA) compared to Rheumatoid Factor-Immunoglobulin M (RF-IgM) and C - reactive protein (CRP). Serum samples of 176 patients were analyzed with the anti-CCP ELISA assay method established in our laboratory. The results of rheumatoid arthritis (RA) patients, the other inflammatory patients, and healthy controls were compared using MedCalc (version 7.0). The anti-CCP assay results were compared with RF-IgM and CRP concentration analyzed in Catholic University. The specificity of ELISA test results of RA patients showed 91% and 87%, when healthy controls or osteoarthritis patients were considered as negative. Thus, the established ELISA method was RA specific, but its sensitivity was low. To see the low sensitivity may from aging effect, the concentration of anti-CCP was analyzed for different aging group. We tested 110 healthy controls' sera using the same method. The statistical results of young subjects (<45 years old) showed significantly lower anti-CCP concentrations than those of older subjects (>65 years old, p<0.0001). The citrullination might also be occurring during aging process in healthy populations. The validation results imply that the established method could be used as a clinical diagnostic for RA together with RF-IgM.
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Effects of sorbitol on porcine oocyte maturation and embryo development in vitro.
Zygote
PUBLISHED: 01-04-2014
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Summary In the present study, a porcine system was supplemented with sorbitol during in vitro maturation (IVM) or in vitro culture (IVC), and the effects of sorbitol on oocyte maturation and embryonic development following parthenogenetic activation were assessed. Porcine immature oocytes were treated with different concentrations of sorbitol during IVM, and the resultant metaphase II stage oocytes were activated and cultured in porcine zygote medium-3 (PZM-3) for 7 days. No significant difference was observed in cumulus expansion and the nuclear maturation between the control and sorbitol-treated groups, with the exception of the 100 mM group, which showed significantly decreased nuclear maturation and cumulus expansion. There was no significant difference in the intracellular reactive oxygen species (ROS) levels between oocytes matured with 10 or 20 mM sorbitol and control groups, but 50 and 100 mM groups had significantly higher ROS levels than other groups. The 20 mM group showed significant increases in intracellular glutathione and subsequent blastocyst formation rates following parthenogenetic activation compared with the other groups. During IVC, supplementation with sorbitol significantly reduced blastocyst formation and increased the apoptotic index compared with the control. The apoptotic index of blastocysts from the sorbitol-treated group for entire culture period was significantly higher than those of the partially sorbitol-exposed groups. Based on these findings, it can be concluded that the addition of a low concentration of sorbitol (20 mM) during IVM of porcine oocytes benefits subsequent blastocyst development and improves embryo quality, whereas sorbitol supplement during IVC has a negative effect on blastocyst formation.
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Development of SPR biosensor for the detection of human hepatitis B virus using plasma-treated parylene-N film.
Biosens Bioelectron
PUBLISHED: 01-03-2014
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A plasma-treated parylene-N film was presented for the immobilization of proteins through physical adsorption. The changes in surface properties of the parylene-N film after plasma-treatment were analyzed using contact angle microscopy and AFM. To demonstrate the high protein-immobilization efficiency of the plasma-treated parylene-N film, the immobilization efficiencies of differently modified surfaces were compared using model proteins with different surface charges, such as streptavidin (pI=5, negatively charged at pH 7), horseradish peroxidase (pI=6.6, nearly neutral at pH 7), and avidin (pI=10, positively charged at pH 7). The application of the plasma-treated parylene-N film as an SPR biosensor was also tested by immobilizing model proteins. An SPR biosensor based on the plasma-treated parylene-N film was developed for the detection of the human hepatitis virus surface antigen (HBsAg), and the plasma-treated parylene-N film was estimated to improve the sensitivity of SPR biosensor as much as 1000-fold by enhancing immobilization of receptor antibodies.
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Polymorphic patterns of the merozoite surface protein-3? in Korean isolates of Plasmodium vivax.
Malar. J.
PUBLISHED: 01-02-2014
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The merozoite surface protein-3? of Plasmodium vivax (PvMSP-3?) is one of the candidate antigens for blood stage malaria vaccine development. The polymorphisms in PvMSP-3? have been reported in certain P. vivax isolates. However, the diversity of PvMSP-3? throughout its global distribution has not been well understood. In this study, the genetic diversity and the effects of natural selection in PvMSP-3? among P. vivax Korean isolates were analysed.
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Striatal cholinergic interneuron regulation and circuit effects.
Front Synaptic Neurosci
PUBLISHED: 01-01-2014
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The striatum plays a central role in motor control and motor learning. Appropriate responses to environmental stimuli, including pursuit of reward or avoidance of aversive experience all require functional striatal circuits. These pathways integrate synaptic inputs from limbic and cortical regions including sensory, motor and motivational information to ultimately connect intention to action. Although many neurotransmitters participate in striatal circuitry, one critically important player is acetylcholine (ACh). Relative to other brain areas, the striatum contains exceptionally high levels of ACh, the enzymes that catalyze its synthesis and breakdown, as well as both nicotinic and muscarinic receptor types that mediate its postsynaptic effects. The principal source of striatal ACh is the cholinergic interneuron (ChI), which comprises only about 1-2% of all striatal cells yet sends dense arbors of projections throughout the striatum. This review summarizes recent advances in our understanding of the factors affecting the excitability of these neurons through acute effects and long term changes in their synaptic inputs. In addition, we discuss the physiological effects of ACh in the striatum, and how changes in ACh levels may contribute to disease states during striatal dysfunction.
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Overexpression of Human GATA-1 and GATA-2 Interferes with Spine Formation and Produces Depressive Behavior in Rats.
PLoS ONE
PUBLISHED: 01-01-2014
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Functional consequences to which vertebrate GATA transcription factors contribute in the adult brain remain largely an open question. The present study examines how human GATA-1 and GATA-2 (hGATA-1 and hGATA-2) are linked to neuronal differentiation and depressive behaviors in rats. We investigated the effects of adeno-associated viral expression of hGATA-1 and hGATA-2 (AAV-hGATA1 and AAV-hGATA2) in the dentate gyrus (DG) of the dorsal hippocampus on dendrite branching and spine number. We also examined the influence of AAV-hGATA1 and AAV-hGATA2 infusions into the dorsal hippocampus on rodent behavior in models of depression. Viral expression of hGATA-1 and hGATA-2 cDNA in rat hippocampal neurons impaired dendritic outgrowth and spine formation. Moreover, viral-mediated expression of hGATA-1 and hGATA-2 in the dorsal hippocampus caused depressive-like deficits in the forced swim test and learned helplessness models of depression, and decreased the expression of several synapse-related genes as well as spine number in hippocampal neurons. Conversely, shRNA knockdown of GATA-2 increased synapse-related gene expression, spine number, and dendrite branching. The results demonstrate that hGATA-1 and hGATA-2 expression in hippocampus is sufficient to cause depressive like behaviors that are associated with reduction in spine synapse density and expression of synapse-related genes.
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Changes in histone H3 lysine 36 methylation in porcine oocytes and preimplantation embryos.
PLoS ONE
PUBLISHED: 01-01-2014
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Histone H3 lysine 36 (H3K36) methylation is known to be associated with transcriptionally active genes, and is considered a genomic marker of active loci. To investigate the changes in H3K36 methylation in pig, we determined the mono-, di-, and tri-methylations of H3K36 (H3K36me1, H3K36me2 and H3K36me3, respectively) in porcine fetal fibroblasts, oocytes and preimplantation embryos by immunocytochemistry using specific antibodies and confocal microscopy. These analyses revealed that only H3K36me3 in porcine fetal fibroblasts consistently colocalized with transcription sites identified as actively synthesizing RNA based on fluorouridine (FU) incorporation. Treatment of cells with flavopiridol, which blocks transcription elongation, completely abrogated both H3K36me3 signals and RNA synthesis. All three types of H3K36 methylation were present and did not significantly differ during oocyte maturation. In parthenogenetic embryos, H3K36me1 and -me2 were detected in 1-cell through blastocyst-stage embryos. In contrast, H3K36me3 was not detected in most 1-cell stage embryos. H3K36me3 signals became detectable in 2-cell stage embryos, peaked at the 4-cell stage, decreased at the 8-cell stage, and then became undetectable at blastocyst stages in both parthenogenetic and in vitro-fertilized (IVF) embryos. Unlike the case in IVF embryos, H3K36me3 could not be demethylated completely during the 1-cell stage in somatic cell nuclear transfer (SCNT) embryos. These results collectively indicate that H3K36me3, but not H3K36me1 or -me2, is associated with transcription elongation in porcine fetal fibroblasts. H3K36me3 is developmentally regulated and may be a histone mark of embryonic gene activation in pig. Aberrant H3K36 tri-methylation occurred during the nuclear reprogramming of SCNT embryos.
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Rare deleterious mutations of the gene EFR3A in autism spectrum disorders.
Mol Autism
PUBLISHED: 01-01-2014
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Whole-exome sequencing studies in autism spectrum disorder (ASD) have identified de novo mutations in novel candidate genes, including the synaptic gene Eighty-five Requiring 3A (EFR3A). EFR3A is a critical component of a protein complex required for the synthesis of the phosphoinositide PtdIns4P, which has a variety of functions at the neural synapse. We hypothesized that deleterious mutations in EFR3A would be significantly associated with ASD.
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Parylene-matrix chip for small molecule analysis using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
Rapid Commun. Mass Spectrom.
PUBLISHED: 01-01-2014
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In matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS), analyte molecules are known to be ionized by mixing with organic matrix molecules. As the organic matrix molecules are ionized, they generate unreproducible mass peaks such that MALDI-TOF MS is nearly impossible in the low mass-to-charge (m/z) range (<1000). In this work, we aimed to develop a parylene-matrix chip for the detection of small molecules in the low m/z range by using MALDI-TOF MS.
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A novel oxiranylchromenone derivative, MHY336, induces apoptosis and cell cycle arrest via a p53- and p21-dependent pathway in HCT116 human colon cancer cells.
Int. J. Oncol.
PUBLISHED: 10-18-2013
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In this study, we compared cytotoxicity, cell cycle distribution, and apoptosis on MHY336 treatment in three human colorectal carcinoma HCT116 cells: p53+/+ (p53?wt), p53-/- (p53-null), and p21-/- (p21-null), as well as investigated the roles of p53 and p21 in cell death. Using these three isogenic variants, the roles of p53 and p21 in the cellular response to treatment with MHY336, a novel topoisomerase II? inhibitor, were investigated. Our results showed that MHY336 treatment increased the expression of p53 over time in cells with wild-type p53 status. This elevated levels of p53 is associated with increased DNA fragmentation, and cleavage of poly(ADP-ribose) polymerase, consistent with increased sensitivity of these cells to apoptotic stimuli. However, p53-null and p21-null cells were more resistant to the antiproliferative and apoptotic effects of MHY336 than p53-wt cells. The same result was achieved by knocking down p53 and p21 with siRNA in p53-wt cells, indicating that p53 and p21 play a crucial role in MHY336-induced cell cycle arrest and apoptosis. Taken together, these results suggest that MHY336 could be a potential candidate to be used in chemoprevention and/or treatment of colon cancer.
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Movement of the ulnar nerve at the elbow: a sonographic study.
J Ultrasound Med
PUBLISHED: 09-26-2013
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The aim of this study was to measure the degree of movement of the ulnar nerve in the cubital tunnel using sonography in patients with ulnar neuropathy at the elbow compared to a healthy control group.
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A novel hydroxamic acid derivative, MHY218, induces apoptosis and cell cycle arrest through downregulation of NF-?B in HCT116 human colon cancer cells.
Int. J. Oncol.
PUBLISHED: 07-22-2013
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Colorectal cancer (CRC) is one of the most common malignant diseases and frequent cause of cancer deaths in the world. In spite of the significant advances in conventional therapeutic approaches to CRC, most patients ultimately die of their disease. There is a need to develop novel preventive approaches for this malignancy. This study was carried out to investigate the anticancer effect of MHY218, a hydroxamic acid derivative, in HCT116 human colon cancer cells. Treatment of cells with MHY218 resulted in growth inhibition and induction of apoptosis in a concentration-dependent manner. MHY218 induced G2/M phase arrest in the cell cycle progression which was observed by flow cytometry analysis, and a decrease in the protein expression of cyclin B1 and its activating partners Cdc25C and Cdc2. MHY218 also caused an increase in the expression levels of p21WAF1/CIP1, a G2/M phase inhibitor, in a p53-independent pathway. The induction of apoptosis was observed by decreased viability, DNA fragmentation, cleavage of poly(ADP-ribose) polymerase, alteration in the ratio of Bax/Bcl-2 protein expression, and activation of caspase-3, -8 and -9. In addition, MHY218 treatment showed downregulation of the expression levels of the transcription factor nuclear factor-kappa B (NF-?B) in the nucleus, which has been reported to be implicated in the apoptotic cell death of several types of cancer cells, suppression of TNF-?-induced NF-?B activation, inhibition of cyclooxygenase-2 expression, repression of matrix metalloproteinase-9 activation and decrease of 5-lipoxygenase in a concentration-dependent manner. These results suggest that MHY218 may be a useful candidate to be used in the chemoprevention and/or treatment of colon cancer.
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Progressive dilation of the left atrium and ventricle after acute myocardial infarction is associated with high mortality.
Korean Circ J
PUBLISHED: 07-10-2013
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The purpose of this study is to identify the prevalence of progressive dilation in patients with acute myocardial infarction (AMI) combined with heart failure (HF) and determine the prognostic significance and associated factors with a geometric change of an infarcted heart.
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Teratogenic potential of antiepileptic drugs in the zebrafish model.
Biomed Res Int
PUBLISHED: 07-09-2013
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The zebrafish model is an attractive candidate for screening of developmental toxicity during early drug development. Antiepileptic drugs (AEDs) arouse concern for the risk of teratogenicity, but the data are limited. In this study, we evaluated the teratogenic potential of seven AEDs (carbamazepine (CBZ), ethosuximide (ETX), valproic acid (VPN), lamotrigine (LMT), lacosamide (LCM), levetiracetam (LVT), and topiramate (TPM)) in the zebrafish model. Zebrafish embryos were exposed to AEDs from initiation of gastrula (5.25 hours post-fertilization (hpf)) to termination of hatching (72?hpf) which mimic the mammalian teratogenic experimental design. The lethality and teratogenic index (TI) of AEDs were determined and the TI values of each drug were compared with the US FDA human pregnancy categories. Zebrafish model was useful screening model for teratogenic potential of antiepilepsy drugs and was in concordance with in vivo mammalian data and human clinical data.
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Identification and characterization of the second cysteine protease inhibitor of Clonorchis sinensis (CsStefin-2).
Parasitol. Res.
PUBLISHED: 07-09-2013
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CsStefin-2, the second cysteine protease inhibitor of Clonorchis sinensis, was identified and characterized. CsStefin-2 is a cysteine protease inhibitor that belongs to family 1 stefins based on its phylogenetic and structural properties. However, CsStefin-2 had a QIVSG cystatin motif distinct from the common QVVAG cystatin motif that is well conserved in family 1 stefins. Mutagenesis analysis revealed that the two amino acid substitutions in the QIVSG cystatin motif of CsStefin-2 did not affect its inhibitory activity. Molecular modeling also indicated that no critical change was induced in the interaction between CsStefin-2 and its target enzyme. CsStefin-2 showed broad inhibitory activities against several cysteine proteases, including human cathepsins B and L, papain, and cathepsin Fs of C. sinensis (CsCFs), and effectively inhibited the autocatalytic maturation of CsCF-6. Native CsStefin-2 was assembled into a homo-tetramer, in which intermolecular disulfide bonds are not involved in the assembly of the tetramer. CsStefin-2 was expressed throughout the various developmental stages of the parasite and was localized in the intestinal epithelium, where CsCFs are actively synthesized. These results suggest that CsStefin-2 is the second active cysteine protease inhibitor of C. sinensis that shares functional redundancy with CsStefin-1 to modulate the activity and processing of CsCFs.
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Baicalein, an active component of Scutellaria baicalensis Georgi, induces apoptosis in human colon cancer cells and prevents AOM/DSS-induced colon cancer in mice.
Int. J. Oncol.
PUBLISHED: 07-09-2013
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Flavonoids have been demonstrated to provide health benefits in humans. Baicalein (5,6,7-trihydroxyflavone) is a phenolic flavonoid compound derived mainly from the root of Scutellaria baicalensis Georgi, a medicinal plant traditionally used in oriental medicine. Baicalein is widely used in Korean and Chinese herbal medicines as anti-inflammatory and anticancer therapy. However, the molecular mechanisms of its activity remain poorly understood and warrant further investigation. This study was performed to investigate the anticancer effect of baicalein on HCT116 human colon cancer cells and the tumor preventing capacity of baicalein on colitis-associated cancer in mice. In in vivo experiments, we induced colon tumors in mice by azoxymethane (AOM) and dextran sulfate sodium (DSS) and evaluated the effects of baicalein on tumor growth. Baicalein treatment on HCT116 cells resulted in a concentration-dependent inhibition of cell growth and induction of apoptotic cell death. The induction of apoptosis was determined by morphological changes and cleavage of poly(ADP-ribose) polymerase. Baicalein also suppressed the activation of NF-?B through PPAR? activation. These results indicate that the anti-inflammatory effects of baicalein may be mediated through PPAR? activation. Finally, administration with baicalein significantly decreased the incidence of tumor formation with inflammation. Our findings suggest that baicalein is one of the candidates for the prevention of inflammation-associated colon carcinogenesis.
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Chamnamul [Pimpinella brachycarpa (Kom.) Nakai] ameliorates hyperglycemia and improves antioxidant status in mice fed a high-fat, high-sucrose diet.
Nutr Res Pract
PUBLISHED: 05-30-2013
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Chronic consumption of a high-fat, high-sucrose (HFHS) diet increases insulin resistance and results in type 2 diabetes mellitus in C57BL/6J mice. Hyperglycemia in diabetics increases oxidative stress, which is associated with a high risk of diabetic complications. The purpose of this study was to examine the hypoglycemic and antioxidant effects of chamnamul [Pimpinella brachycarpa (Kom.) Nakai] in an animal model of type 2 diabetes. The ?-glucosidase inhibitory activity of a 70% ethanol extract of chamnamul was measured in vitro. Five-week-old male C57BL/6J mice were fed a basal or HFHS diet with or without a 70% ethanol extract of chamnamul at a 0.5% level of the diet for 12 weeks after 1 week of adaptation. After sacrifice, serum glucose, insulin, adiponectin, and lipid profiles, and lipid peroxidation of the liver were determined. Homeostasis model assessment for insulin resistance (HOMA-IR) was determined. Chamnamul extract inhibited ?-glucosidase by 26.7%, which was 78.3% the strength of inhibition by acarbose at a concentration of 0.5 mg/mL. Serum glucose, insulin, and cholesterol levels, as well as HOMA-IR values, were significantly lower in the chamnamul group than in the HFHS group. Chamnamul extract significantly decreased the level of thiobarbituric acid reactive substances and increased the activities of superoxide dismutase, catalase, and glutathione peroxidase in the liver compared with the HFHS group. These findings suggest that chamnamul may be useful in prevention of hyperglycemia and reduction of oxidative stress in mice fed a HFHS diet.
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Predicting need for fixation of atypical femoral fracture.
J. Clin. Endocrinol. Metab.
PUBLISHED: 05-13-2013
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Predictors of the requirement for fixation have not been reported in incomplete atypical femoral fractures. The clinical features of incomplete atypical femoral fractures should be reviewed to predict the requirement for surgical intervention in this condition.
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Comparison of Coronary Plaque Components between Non-Culprit Lesions in Patients with Acute Coronary Syndrome and Target Lesions in Patients with Stable Angina: Virtual Histology-Intravascular Ultrasound Analysis.
Korean Circ J
PUBLISHED: 04-28-2013
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The differences in plaque characteristics between non-culprit lesions (NCL) in acute coronary syndrome (ACS) patients (ACS-NCL) and target lesions (TL) in stable angina (SA) patients (SA-TL) are not well understood. We used a virtual histology-intravascular ultrasound (VH-IVUS) to compare the plaque components between ACS-NCL and SA-TL.
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Defining the regulatory and inhibitory elements within the prodomain of CsCF-6, a cathepsin F cysteine protease of Clonorchis sinensis.
Mol. Biochem. Parasitol.
PUBLISHED: 04-20-2013
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CsCF-6 is a member of the multigene family of cathepsin F cysteine proteases of Clonorchis sinensis. Similar to other papain family proteases, CsCF-6 is synthesized as a proenzyme and is converted to the mature form by autocatalytic removal of the prodomain. Here, we analyzed the regulatory and inhibitory elements within the CsCF-6 prodomain to understand the regulatory mechanism of CsCF-6 by its prodomain. The CsCF-6 prodomain played an essential role in the folding of CsCF-6. Particularly, the ERFNAQ motif within the prodomain was essential, and the minimum segment required for this event was the C-terminal part of the prodomain, including Asn(58) and downstream residues. The CsCF-6 prodomain effectively inhibited CsCF-6, in which the ERFNAQ motif played a critical role in the inhibition, but the GTFD motif was also required for complete inhibition of CsCF-6. The CsCF-6 prodomain showed broad inhibitory activity against several cysteine proteases. These results suggest that the CsCF-6 prodomain plays bi-functional roles in correct folding and inhibition of its cognate enzyme.
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The role of neuroplasticity in dopaminergic therapy for Parkinson disease.
Nat Rev Neurol
PUBLISHED: 04-16-2013
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Dopamine replacement is a mainstay of therapeutic strategies for Parkinson disease (PD). The motor response to therapy involves an immediate improvement in motor function, known as the short-duration response (SDR), followed by a long-duration response (LDR) that develops more slowly, over weeks. Here, we review evidence in patients and animal models suggesting that dopamine-dependent corticostriatal plasticity, and retention of such plasticity in the absence of dopamine, are the mechanisms underlying the LDR. Conversely, experience-dependent aberrant plasticity that develops slowly under reduced dopamine levels could contribute substantially to PD motor symptoms before initiation of dopamine replacement therapy. We place these findings in the context of the role of dopamine in basal ganglia function and corticostriatal plasticity, and provide a new framework suggesting that therapies that enhance the LDR could be more effective than those targeting the SDR. We further propose that changes in neuroplasticity constitute a form of disease modification that is distinct from prevention of degeneration, and could be responsible for some of the unexplained disease-modifying effects of certain therapies. Understanding such plasticity could provide novel therapeutic approaches that combine rehabilitation and pharmacotherapy for treatment of neurological and psychiatric disorders involving basal ganglia dysfunction.
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Longitudinal assessment of tau and amyloid beta in cerebrospinal fluid of Parkinson disease.
Acta Neuropathol.
PUBLISHED: 04-01-2013
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Tau gene has been consistently associated with the risk of Parkinson disease in recent genome wide association studies. In addition, alterations of the levels of total tau, phosphorylated tau [181P], and amyloid beta 1-42 in cerebrospinal fluid have been reported in patients with sporadic Parkinson disease and asymptomatic carriers of leucine-rich repeat kinase 2 mutations, in patterns that clearly differ from those typically described for patients with Alzheimer disease. To further determine the potential roles of these molecules in Parkinson disease pathogenesis and/or in tracking the disease progression, especially at early stages, the current study assessed all three proteins in 403 Parkinson disease patients enrolled in the DATATOP (Deprenyl and tocopherol antioxidative therapy of parkinsonism) placebo-controlled clinical trial, the largest cohort to date with cerebrospinal fluid samples collected longitudinally. These initially drug-naive patients at early disease stages were clinically evaluated, and cerebrospinal fluid was collected at baseline and then at endpoint, defined as the time at which symptomatic anti-Parkinson disease medications were determined to be required. General linear models were used to test for associations between baseline cerebrospinal fluid biomarker levels or their rates of change and changes in the Unified Parkinson Disease Rating Scale (total or part III motor score) over time. Robust associations among candidate markers are readily noted. Baseline levels of amyloid beta were weakly but negatively correlated with baseline Unified Parkinson Disease Rating Scale total scores. Baseline phosphorylated tau/total tau and phosphorylated tau/amyloid beta were significantly and negatively correlated with the rates of the Unified Parkinson Disease Rating Scale change. While medications (deprenyl and/or tocopherol) did not appear to alter biomarkers appreciably, a weak but significant positive correlation between the rate of change in total tau or total tau/amyloid beta levels and the change of the Unified Parkinson Disease Rating Scale was observed. Notably, these correlations did not appear to be influenced by APOE genotype. These results are one of the very first pieces of evidence suggesting that tau and amyloid beta are critically involved in early Parkinson disease progression, potentially by a different mechanism than that in Alzheimer disease, although their applications as Parkinson disease progression markers will likely require the addition of other proteins.
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Effect of Gambisan on the Inhibition of Adipogenesis in 3T3-L1 Adipocytes.
Evid Based Complement Alternat Med
PUBLISHED: 03-26-2013
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This study was conducted to explore the antiadipogenic effect and possible mechanism of Gambisan on 3T3-L1 cells. For quality control, Gambisan was standardized by HPLC and the standard compounds ephedrine, epigallocatechin-3-gallate, and caffeine were screened. Cultured 3T3-L1 cells that had been induced to differentiate were treated with various concentrations of Gambisan or its major component extracts (Ephedra intermedia Schrenk, Atractylodes lancea DC., and Thea sinensis L.) for 72 hours for MTT assay to determine cell viability or 10 days for LDH assay, triglyceride assay, DNA content measurement, Oil red O staining, RT-PCR, and western blot. Gambisan significantly inhibited adipogenesis in 3T3-L1 cells by reducing triglyceride contents and lipid accumulation in a dose-dependent manner without obvious cytotoxicity. Viability and DNA content in 3T3-L1 cells treated with Gambisan were significantly higher than cells treated with the major component extracts at every concentration. The anti-adipogenic effects of Gambisan appeared to be mediated by a significant downregulation of the expression of lipoprotein lipase mRNA and PPAR ? , C/EBP ? , and SREBP-1 protein apart from the expression of hormone-sensitive lipase. Gambisan could act as a possible therapeutic agent for obesity. However, further studies including in vivo assays and clinical trials are needed to confirm the efficacy, safety and mechanisms of the antiobesity effects of Gambisan.
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Magnetic-bead-based immunoassay using E. coli cells with autodisplayed Z-domains.
Enzyme Microb. Technol.
PUBLISHED: 03-22-2013
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Escherichia coli cells with autodisplayed Z-domains could increase the sensitivity of immunoassays by immobilizing antibodies in a controlled orientation. In the work presented here, E. coli cells with autodisplayed Z-domains were immobilized to magnetic beads for subsequent immunoassay. In comparing conventional immunoassay using the E. coli cells with autodisplayed Z-domains, the magnetic-bead-based immunoassay improved immunoassay efficiency by minimizing the loss of E. coli cells during repeated centrifugation steps during washing. For the immobilization of E. coli cells to magnetic beads, the magnetic beads were modified with poly-l-lysine to bind to negatively charged E. coli cells. During the surface modification process, physical parameters such as the surface charge and size of the magnetic beads were analyzed to confirm the formation of E. coli-magnetic bead complexes. To test the feasibility of the magnetic-bead-based immunoassay, horseradish peroxidase (HRP) was used as a model analyte, and a biomarker for inflammatory diseases, C-reactive protein (CRP), was used for a demonstration of an application in medical diagnosis.
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Flow cytometric immunoassay using E. coli with autodisplayed Z-domains.
Enzyme Microb. Technol.
PUBLISHED: 03-05-2013
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Recently, we reported a highly sensitive immunoassay using Escherichia coli cells with autodisplayed Z-domains. In this work, E. coli cells with autodisplayed Z-domains were applied to the flow-cytometry-based simultaneous detection of multiple analytes. The E. coli cells were doubly transfected to express a fluorescent protein (tdTomato) in the cytosol and the autodisplayed Z-domains on the outer membrane. By using E. coli cells with only the autodisplayed Z-domains, immunoassay of multiple analytes could be performed simultaneously on the same sample. Flow cytometry can be used to identify the immunoassay type by simultaneously detecting the fluorescence signal from the cytosol (tdTomato) and the fluorescence from the outer membrane, enabling the quantification of bound analytes after treatment with additional fluorescently labeled antibodies. To demonstrate the immunoassay of multiple analytes by using flow cytometry, human hepatitis B virus surface antigen (HBsAg) and C-reactive protein (CRP), a broad spectrum inflammation marker, were used as model analytes.
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Moxibustion for treating knee osteoarthritis: study protocol of a multicentre randomised controlled trial.
BMC Complement Altern Med
PUBLISHED: 02-28-2013
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The treatment of knee osteoarthritis, which is a major cause of disability among the elderly, is typically selected from multidisciplinary options, including complementary and alternative medicine. Moxibustion has been used in the treatment of knee osteoarthritis in Korea to reduce pain and improve physical activity. However, there is no sufficient evidence of its effectiveness, and it cannot therefore be widely recommended for treating knee osteoarthritis. We designed a randomised controlled clinical trial to evaluate the effectiveness, safety, cost-effectiveness, and qualitative characteristics of moxibustion treatment of knee osteoarthritis compared to usual care.
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