JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
Sonographic appearances of juvenile fibroadenoma of the breast.
J Ultrasound Med
PUBLISHED: 10-23-2014
Show Abstract
Hide Abstract
The purpose of this study was to evaluate characteristic features of juvenile fibroadenoma of the breast on sonography.
Related JoVE Video
Rugged and breathable forms of stretchable electronics with adherent composite substrates for transcutaneous monitoring.
Nat Commun
PUBLISHED: 09-03-2014
Show Abstract
Hide Abstract
Research in stretchable electronics involves fundamental scientific topics relevant to applications with importance in human healthcare. Despite significant progress in active components, routes to mechanically robust construction are lacking. Here, we introduce materials and composite designs for thin, breathable, soft electronics that can adhere strongly to the skin, with the ability to be applied and removed hundreds of times without damaging the devices or the skin, even in regions with substantial topography and coverage of hair. The approach combines thin, ultralow modulus, cellular silicone materials with elastic, strain-limiting fabrics, to yield a compliant but rugged platform for stretchable electronics. Theoretical and experimental studies highlight the mechanics of adhesion and elastic deformation. Demonstrations include cutaneous optical, electrical and radio frequency sensors for measuring hydration state, electrophysiological activity, pulse and cerebral oximetry. Multipoint monitoring of a subject in an advanced driving simulator provides a practical example.
Related JoVE Video
Effect of oral Administration of Lactobacillus plantarum HY7714 on Epidermal Hydration in Ultraviolet B-Irradiated Hairless Mice.
J. Microbiol. Biotechnol.
PUBLISHED: 09-01-2014
Show Abstract
Hide Abstract
In recent years, probiotics have gained interest owing to their beneficial effects on skin health, such as inhibiting atopic dermatitis and improving skin immunity or preventing inflammation. However, little is known about the effects of probiotics on ultraviolet B (UVB)-induced reduction in skin hydration. In this study, we evaluated the effect of Lactobacillus plantarum HY7714 on skin hydration in human dermal fibroblasts and in hairless mice. In Hs68 cells, L. plantarum HY7714 not only increased serine palmitoyltransferase (SPT) mRNA level, but also decreased ceramidase mRNA level. In order to confirm the hydrating effects of L. plantarum HY7714 in vivo, we orally administered vehicle or L. plantarum HY7714 at a dose of 1 × 109 CFU/day to hairless mice for 8 weeks. In hairless mice, L. plantarum HY7714 significantly decreased UVB-induced epidermal thickness. In addition, we found that L. plantarum HY7714 administration significantly suppressed the increase in transepidermal water loss (TEWL) and decrease in skin hydration, which reflects barrier function fluctuations following UV irradiation. In particular, L. plantarum HY7714 administration significantly increased the ceramide level compared to that in the UVB group. In the experiment on SPT and ceramidase mRNA expressions, L. plantarum HY7714 administration significantly improved the reduction in SPT mRNA levels and suppressed the increase in ceramidase mRNA levels caused by UVB in the hairless mice skins. Collectively, these results provide further insight that L. plantarum HY7714 can be a potential candidate for preserving skin hydration levels against UV irradiation.
Related JoVE Video
Cytotoxic activity of 3,6-dihydroxyflavone in human cervical cancer cells and its therapeutic effect on c-Jun N-terminal kinase inhibition.
Molecules
PUBLISHED: 08-27-2014
Show Abstract
Hide Abstract
Previously we have shown that 3,6-dihydroxyflavone (3,6-DHF) is a potent agonist of the human peroxisome proliferator-activated receptor (hPPAR) with cytotoxic effects on human cervical cancer cells. To date, the mechanisms by which 3,6-DHF exerts its antitumor effects on cervical cells have not been clearly defined. Here, we demonstrated that 3,6-DHF exhibits a novel antitumor activity against HeLa cells with IC50 values of 25 ?M and 9.8 ?M after 24 h and 48 h, respectively. We also showed that the anticancer effects of 3,6-DHF are mediated via the toll-like receptor (TLR) 4/CD14, p38 mitogen-activated protein kinase (MAPK), Jun-N terminal kinase (JNK), extracellular-signaling regulated kinase (ERK), and cyclooxygenase (COX)-2 pathways in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. We found that 3,6-DHF showed a similar IC50 (113 nM) value to that of the JNK inhibitor, SP600125 (IC50 = 118 nM) in a JNK1 kinase assay. Binding studies revealed that 3,6-DHF had a strong binding affinity to JNK1 (1.996 × 105 M-1) and that the 6-OH and the carbonyl oxygen of the C ring of 3,6-DHF participated in hydrogen bonding interactions with the carbonyl oxygen and the amide proton of Met111, respectively. Therefore, 3,6-DHF may be a candidate inhibitor of JNKs, with potent anticancer effects.
Related JoVE Video
Ultrasonographic findings of Kimura's disease presenting in the upper extremities.
Jpn J Radiol
PUBLISHED: 08-05-2014
Show Abstract
Hide Abstract
To describe ultrasound findings of Kimura's disease arising in the upper extremities.
Related JoVE Video
Annual Change in Pulmonary Function and Clinical Characteristics of Combined Pulmonary Fibrosis and Emphysema and Idiopathic Pulmonary Fibrosis: Over a 3-Year Follow-up.
Tuberc Respir Dis (Seoul)
PUBLISHED: 07-31-2014
Show Abstract
Hide Abstract
Combined pulmonary fibrosis and emphysema (CPFE) have different pulmonary function tests (PFTs) and outcomes than idiopathic pulmonary fibrosis (IPF). The intention of this study was to identify unknown differences between CPFE and IPF by a retrospective comparison of clinical data including baseline and annual changes in pulmonary function, comorbidities, laboratory findings, clinical characteristics and cause of hospitalization.
Related JoVE Video
Structure and flexibility of the thermophilic cold-shock protein of Thermus aquaticus.
Biochem. Biophys. Res. Commun.
PUBLISHED: 07-21-2014
Show Abstract
Hide Abstract
The thermophilic bacterium Thermus aquaticus is a well-known source of Taq polymerase. Here, we studied the structure and dynamics of the T. aquaticus cold-shock protein (Ta-Csp) to better understand its thermostability using NMR spectroscopy. We found that Ta-Csp has a five-stranded ?-barrel structure with five salt bridges which are important for more rigid structure and a higher melting temperature (76 °C) of Ta-Csp compared to mesophilic and psychrophilic Csps. Microsecond to millisecond time scale exchange processes occur only at the ?1-?2 surface region of the nucleic acid binding site with an average conformational exchange rate constant of 674 s(-1). The results imply that thermophilic Ta-Csp has a more rigid structure and may not need high structural flexibility to accommodate nucleic acids upon cold shock compared to its mesophile and psychrophile counterparts.
Related JoVE Video
Ultrasonographic features of vascular closure devices: initial and 6-month follow-up results.
Ultrasonography
PUBLISHED: 07-16-2014
Show Abstract
Hide Abstract
This study aimed to evaluate the ultrasonographic findings for various types of vascular closure devices (VCDs) immediately after the angiographic procedure and at 6-month follow-up.
Related JoVE Video
Structure and backbone dynamics of vanadate-bound PRL-3: comparison of 15N nuclear magnetic resonance relaxation profiles of free and vanadate-bound PRL-3.
Biochemistry
PUBLISHED: 07-14-2014
Show Abstract
Hide Abstract
Phosphatases of regenerating liver (PRLs) constitute a novel class of small, prenylated phosphatases with oncogenic activity. PRL-3 is particularly important in cancer metastasis and represents a potential therapeutic target. The flexibility of the WPD loop as well as the P-loop of protein tyrosine phosphatases is closely related to their catalytic activity. Using nuclear magnetic resonance spectroscopy, we studied the structure of vanadate-bound PRL-3, which was generated by addition of sodium orthovanadate to PRL-3. The WPD loop of free PRL-3 extended outside of the active site, forming an open conformation, whereas that of vanadate-bound PRL-3 was directed into the active site by a large movement, resulting in a closed conformation. We suggest that vanadate binding induced structural changes in the WPD loop, P-loop, helices ?4-?6, and the polybasic region. Compared to free PRL-3, vanadate-bound PRL-3 has a longer ?4 helix, where the catalytic R110 residue coordinates with vanadate in the active site. In addition, the hydrophobic cavity formed by helices ?4-?6 with a depth of 14-15 Å can accommodate a farnesyl chain at the truncated prenylation motif of PRL-3, i.e., from R169 to M173. Conformational exchange data suggested that the WPD loop moves between open and closed conformations with a closing rate constant k(close) of 7 s(-1). This intrinsic loop flexibility of PRL-3 may be related to their catalytic rate and may play a role in substrate recognition.
Related JoVE Video
Deltoid muscle injury scratched by an osteophyte on the greater tuberosity of the humeral head: an unusual cause of shoulder pain.
Knee Surg Sports Traumatol Arthrosc
PUBLISHED: 06-03-2014
Show Abstract
Hide Abstract
Few reports to date have discussed acute deltoid muscle injury, including partial or complete deltoid muscle detachment from its origin. The present report describes a different pathomechanism and clinical manifestations of deltoid muscle injury. A 52-year-old man experienced acute severe pain in the right shoulder as a result of abrupt elevation of the arm. The deltoid muscle had apparently been scratched by an osteophyte on the greater tuberosity. After arthroscopic excision of the osteophyte, the symptoms resolved completely. Level of evidence Case report with no comparison group, Level IV.
Related JoVE Video
A phase II trial of Cremorphor EL-free paclitaxel (Genexol-PM) and gemcitabine in patients with advanced non-small cell lung cancer.
Cancer Chemother. Pharmacol.
PUBLISHED: 05-23-2014
Show Abstract
Hide Abstract
Genexol-PM is a Cremorphor EL (CrEL)-free polymeric micelle formulation of paclitaxel that allows higher-dose administration with less hypersensitivity. This study was designed to evaluate the efficacy and safety of Genexol-PM and gemcitabine combination in advanced non-small cell lung cancer patients as a first-line treatment.
Related JoVE Video
Ordered packing of emulsion droplets toward the preparation of adjustable photomasks.
Langmuir
PUBLISHED: 05-08-2014
Show Abstract
Hide Abstract
Monodisperse emulsion droplets with a high volume fraction form crystalline phases that can potentially serve as adjustable photomasks in photolithography. Such photomasks were prepared using a microfluidic device in which a flow-focusing junction, side channels, and a reservoir were connected in series. Transparent oil droplets were generated in a dye-containing continuous water phase at the flow-focusing junction. The droplets were then concentrated through the selective removal of the continuous phase using the side channels. This process led to the formation of a regular array of droplets in the reservoir with a configuration that depended on the relative height of the reservoir to the droplet diameter. The configurations could be selected among a single-layered hexagonal array, a bilayered square array, and a bilayered hexagonal array. The droplet arrays were used as a photomask to create hexagonal or square arrays of microdots. The transmittance profile of the ultraviolet (UV) light from each droplet was parabolic, which enabled the dot size to be tuned by controlling the UV irradiation time. This mask effect is otherwise difficult to achieve using conventional photomasks. The dot size and array periodicity could be adjusted by the in-situ control of the droplet size at the flow-focusing droplet maker. The combination of droplet size adjustments and the UV irradiation time provided independent control over the dot size and array periodicity to enable the preparation of a series of hexagonal microarrays with a wide spectrum of array parameters using a single microfluidic device.
Related JoVE Video
Ultrasound elastography of lateral epicondylosis: clinical feasibility of quantitative elastographic measurements.
AJR Am J Roentgenol
PUBLISHED: 04-25-2014
Show Abstract
Hide Abstract
The objective of our study was to investigate the clinical feasibility of ultrasound elastography for assessing patients with lateral epicondylosis and to establish an objective and quantitative method of elastographic measurement in the affected tendon.
Related JoVE Video
Brain MRI in neuropsychiatric lupus: associations with the 1999 ACR case definitions.
Rheumatol. Int.
PUBLISHED: 04-09-2014
Show Abstract
Hide Abstract
The purpose of this study was to identify the characteristic magnetic resonance imaging (MRI) findings in neuropsychiatric systemic lupus erythematosus (NPSLE) and to investigate the association between MRI findings and neuropsychiatric manifestations in SLE. Brain MRIs with a diagnosis of SLE from 2002 to 2013 from three tertiary university hospitals were screened. All clinical manifestations evaluated by brain MRI were retrospectively reviewed. If the clinical manifestations were compatible with the 1999 NPSLE American College of Rheumatology (ACR) nomenclature and case definitions, the brain MRIs were assessed for the presence of white matter hyperintensities, gray matter hyperintensities, parenchymal defects, atrophy, enhancement, and abnormalities in diffusion-weighted images (DWI). The number, size, and location of each lesion were evaluated. The neuropsychiatric manifestation of each brain MRI was classified according to the 1999 ACR NPSLE case definitions. The associations between MRI findings and NPSLE manifestations were examined. In total, 219 brain MRIs with a diagnosis of SLE were screened, and 133 brain MRIs met the inclusion criteria for NPSLE. The most common MRI abnormality was white matter hyperintensities, which were observed in 76 MRIs (57.1 %). Gray matter hyperintensities were observed in 41 MRIs (30.8 %). Parenchymal defects were found in 31 MRIs (23.3 %), and atrophy was detected in 20 MRIs (15.0 %). Patients who had seizures were more associated with gray matter hyperintensities than patients with other neuropsychiatric manifestations. Patients with cerebrovascular disease were more associated with gray matter hyperintensity, parenchymal defects, and abnormal DWI than patients with other neuropsychiatric manifestations. In addition to white matter hyperintensities, which were previously known as SLE findings, we also noted the presence of gray matter hyperintensities, parenchymal defects, and abnormal DWI in a substantial portion of SLE patients, particularly in those with cerebrovascular disease or seizures.
Related JoVE Video
Tyrosine-mediated two-dimensional peptide assembly and its role as a bio-inspired catalytic scaffold.
Nat Commun
PUBLISHED: 03-14-2014
Show Abstract
Hide Abstract
In two-dimensional interfacial assemblies, there is an interplay between molecular ordering and interface geometry, which determines the final morphology and order of entire systems. Here we present the interfacial phenomenon of spontaneous facet formation in a water droplet driven by designed peptide assembly. The identified peptides can flatten the rounded top of a hemispherical droplet into a plane by forming a macroscopic two-dimensional crystal structure. Such ordering is driven by the folding geometry of the peptide, interactions of tyrosine and crosslinked stabilization by cysteine. We discover the key sequence motifs and folding structures and study their sequence-specific assembly. The well-ordered, densely packed, redox-active tyrosine units in the YYACAYY (H-Tyr-Tyr-Ala-Cys-Ala-Tyr-Tyr-OH) film can trigger or enhance chemical/electrochemical reactions, and can potentially serve as a platform to fabricate a molecularly tunable, self-repairable, flat peptide or hybrid film.
Related JoVE Video
Comparison of sputum and nasopharyngeal swabs for detection of respiratory viruses.
J. Med. Virol.
PUBLISHED: 03-04-2014
Show Abstract
Hide Abstract
Diagnostic tests for respiratory viral infections use traditionally either nasopharyngeal washes or swabs. Sputum is representative of the lower respiratory tract but is used rarely for viral testing. The aim of this study was to compare the detection rates of respiratory viruses from nasopharyngeal swabs and sputum using a multiplex real-time reverse transcription-polymerase chain reaction (RT-PCR). Adults who were admitted or presented to the clinics of Gil Medical Center with acute respiratory symptoms were recruited from 1 November 2012 to 31 March 2013. Paired specimens of nasopharyngeal swabs and sputum were obtained from 154 subjects, and RNA was extracted and tested for 16 different respiratory viruses using the Anyplex II RV16 Detection kit (Seegene, Seoul, Korea). The positive rate was 53% (81/154) for nasopharyngeal swabs and 68% (105/154) for sputum (P?
Related JoVE Video
Preoperative embolization of cerebellar hemangioblastoma with onyx: report of three cases.
Neurointervention
PUBLISHED: 02-28-2014
Show Abstract
Hide Abstract
Hemangioblastoma is a benign and highly vascular tumor. Complete surgical resection of highly vascular tumor such as hemangioblastoma may be challenging due to excessive bleeding. Preoperative embolization of these lesions may decrease the intraoperative blood loss and facilitate excision. We report three cases of cerebellar hemangioblastomas that were embolized using Onyx.
Related JoVE Video
Multibanded anterior talofibular ligaments in normal ankles and sprained ankles using 3D isotropic proton density-weighted fast spin-echo MRI sequence.
AJR Am J Roentgenol
PUBLISHED: 02-27-2014
Show Abstract
Hide Abstract
The purpose of this study is to evaluate the frequency and MRI characteristics of multibanded anterior talofibular ligaments in normal ankles and to characterize the tear types of multibanded anterior talofibular ligaments in sprained ankles using a 3D isotropic proton density (PD)-weighted fast spin-echo (FSE) MRI sequence.
Related JoVE Video
Multifunctional skin-like electronics for quantitative, clinical monitoring of cutaneous wound healing.
Adv Healthc Mater
PUBLISHED: 02-02-2014
Show Abstract
Hide Abstract
Non-invasive, biomedical devices have the potential to provide important, quantitative data for the assessment of skin diseases and wound healing. Traditional methods either rely on qualitative visual and tactile judgments of a professional and/or data obtained using instrumentation with forms that do not readily allow intimate integration with sensitive skin near a wound site. Here, an electronic sensor platform that can softly and reversibly laminate perilesionally at wounds to provide highly accurate, quantitative data of relevance to the management of surgical wound healing is reported. Clinical studies on patients using thermal sensors and actuators in fractal layouts provide precise time-dependent mapping of temperature and thermal conductivity of the skin near the wounds. Analytical and simulation results establish the fundamentals of the sensing modalities, the mechanics of the system, and strategies for optimized design. The use of this type of "epidermal" electronics system in a realistic clinical setting with human subjects establishes a set of practical procedures in disinfection, reuse, and protocols for quantitative measurement. The results have the potential to address important unmet needs in chronic wound management.
Related JoVE Video
3D multifunctional integumentary membranes for spatiotemporal cardiac measurements and stimulation across the entire epicardium.
Nat Commun
PUBLISHED: 01-27-2014
Show Abstract
Hide Abstract
Means for high-density multiparametric physiological mapping and stimulation are critically important in both basic and clinical cardiology. Current conformal electronic systems are essentially 2D sheets, which cannot cover the full epicardial surface or maintain reliable contact for chronic use without sutures or adhesives. Here we create 3D elastic membranes shaped precisely to match the epicardium of the heart via the use of 3D printing, as a platform for deformable arrays of multifunctional sensors, electronic and optoelectronic components. Such integumentary devices completely envelop the heart, in a form-fitting manner, and possess inherent elasticity, providing a mechanically stable biotic/abiotic interface during normal cardiac cycles. Component examples range from actuators for electrical, thermal and optical stimulation, to sensors for pH, temperature and mechanical strain. The semiconductor materials include silicon, gallium arsenide and gallium nitride, co-integrated with metals, metal oxides and polymers, to provide these and other operational capabilities. Ex vivo physiological experiments demonstrate various functions and methodological possibilities for cardiac research and therapy.
Related JoVE Video
Anti-inflammatory activity of rhamnetin and a model of its binding to c-Jun NH2-terminal kinase 1 and p38 MAPK.
J. Nat. Prod.
PUBLISHED: 01-07-2014
Show Abstract
Hide Abstract
Rhamnetin (1), a commonly occurring plant O-methylated flavonoid, possesses antioxidant properties. To address the potential therapeutic efficacy of 1, its anti-inflammatory activity and mode of action in mouse macrophage-derived RAW264.7 cells stimulated with lipopolysaccharide (LPS) or interferon (IFN)-? were investigated. Rhamnetin (1) suppressed mouse tumor necrosis factor (mTNF)-?, mouse macrophage inflammatory protein (mMIP)-1, and mMIP-2 cytokine production in LPS-stimulated macrophages. A nontoxic dose of 1 suppressed nitric oxide production. It was found that the anti-inflammatory effects of 1 are mediated by actions on the p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and cyclooxygenase (COX)-2 pathways in LPS- or IFN-?-stimulated RAW264.7 cells. It was determined that 1 binds to human JNK1 (9.7 × 10(8) M(-1)) and p38 MAPK (2.31 × 10(7) M(-1)) with good affinity. The binding model showed interactions with the 3'- and 4'-hydroxy groups of the B-ring and the 5-hydroxy group of the A-ring of 1. Further, 1 exerted an anti-inflammatory effect, reducing the levels of pro-inflammatory cytokines and mediators.
Related JoVE Video
SMILE inhibits BMP-2-induced expression of osteocalcin by suppressing the activity of the RUNX2 transcription factor in MC3T3E1 cells.
Bone
PUBLISHED: 01-02-2014
Show Abstract
Hide Abstract
Small heterodimer partner interacting leucine zipper protein (SMILE) is an orphan nuclear receptor and a member of the bZIP family of proteins. Several recent studies have suggested that SMILE is a novel co-repressor that is involved in nuclear receptor signaling; however, the role of SMILE in osteoblast differentiation has not yet been elucidated. This study demonstrates that SMILE inhibits osteoblast differentiation by regulating the activity of Runt-related transcription factor-2 (RUNX2). Tunicamycin, an inducer of endoplasmic reticulum stress, stimulated SMILE expression. Bone morphogenetic protein-2-induced expression of alkaline phosphatase and osteocalcin, both of which are osteogenic genes, was suppressed by SMILE. The molecular mechanism by which SMILE affects osteocalcin expression was also determined. An immunoprecipitation assay revealed a physical interaction between SMILE and RUNX2 that significantly impaired the RUNX2-dependent activation of the osteocalcin gene. A ChIP assay revealed that SMILE repressed the ability of RUNX2 to bind to the osteocalcin gene promoter. Taken together, these findings demonstrate that SMILE negatively regulates osteocalcin via a direct interaction with RUNX2.
Related JoVE Video
Investigation of de novo unique differentially expressed genes related to evolution in exercise response during domestication in Thoroughbred race horses.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Previous studies of horse RNA-seq were performed by mapping sequence reads to the reference genome during transcriptome analysis. However in this study, we focused on two main ideas. First, differentially expressed genes (DEGs) were identified by de novo-based analysis (DBA) in RNA-seq data from six Thoroughbreds before and after exercise, here-after referred to as "de novo unique differentially expressed genes" (DUDEG). Second, by integrating both conventional DEGs and genes identified as being selected for during domestication of Thoroughbred and Jeju pony from whole genome re-sequencing (WGS) data, we give a new concept to the definition of DEG. We identified 1,034 and 567 DUDEGs in skeletal muscle and blood, respectively. DUDEGs in skeletal muscle were significantly related to exercise-induced stress biological process gene ontology (BP-GO) terms: 'immune system process'; 'response to stimulus'; and, 'death' and a KEGG pathways: 'JAK-STAT signaling pathway'; 'MAPK signaling pathway'; 'regulation of actin cytoskeleton'; and, 'p53 signaling pathway'. In addition, we found TIMELESS, EIF4A3 and ZNF592 in blood and CHMP4C and FOXO3 in skeletal muscle, to be in common between DUDEGs and selected genes identified by evolutionary statistics such as FST and Cross Population Extended Haplotype Homozygosity (XP-EHH). Moreover, in Thoroughbreds, three out of five genes (CHMP4C, EIF4A3 and FOXO3) related to exercise response showed relatively low nucleotide diversity compared to the Jeju pony. DUDEGs are not only conceptually new DEGs that cannot be attained from reference-based analysis (RBA) but also supports previous RBA results related to exercise in Thoroughbred. In summary, three exercise related genes which were selected for during domestication in the evolutionary history of Thoroughbred were identified as conceptually new DEGs in this study.
Related JoVE Video
Advanced glycation end-products and receptor for advanced glycation end-products expression in patients with idiopathic pulmonary fibrosis and NSIP.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Advanced glycation end products (AGEs) are associated with the pathogenesis of various diseases. AGEs induce excess accumulation of extracellular matrix and expression of profibrotic cytokines. In addition, studies on receptor for advanced glycation end products (RAGE) have shown that the ligand-RAGE interaction activates several intracellular signaling cascades associated with several fibrotic diseases. We investigated the expression of AGEs and RAGE in samples from patients with idiopathic pulmonary fibrosis (IPF) and non-specific interstitial pneumonia (NSIP). Lung tissues and plasma samples from patients with IPF (n=10), NSIP (n=10), and control subjects (n=10) were obtained. Expression of AGEs and RAGE was determined by immunofluorescence assay of lung tissue. Circulating AGEs were measured by Western blot and enzyme-linked immunosorbent assay. Lungs with IPF showed strong expression for both AGEs and RAGE compared to that in NSIP and controls. However, no difference in AGE or RAGE expression was observed in lungs with NSIP compared to that in the controls. Levels of circulating AGEs also increased significantly in lungs of patients with IPF compared to those with NSIP and normal control. Increased AGE-RAGE interaction may play an important role in the pathogenesis of IPF.
Related JoVE Video
Relationship between structural flexibility and function in the C-terminal region of the heparin-binding domain of VEGF165.
Biochemistry
PUBLISHED: 11-27-2013
Show Abstract
Hide Abstract
Vascular endothelial growth factor (VEGF) is an angiogenic protein with neurotrophic and neuroprotective effects. Previously, we reported that triamterene (Trm) inhibits VEGF-A? (amyloid ?) interactions without affecting other biological activities of VEGF or A? [Jeong, K-W. et al. (2011) Biochemistry 50, 4843-4854]. We further showed that molecular motions in the N-terminal disordered loop region of the heparin-binding domain (HBD) are important for interaction with Trm. To investigate the importance of motion at the C-terminal domain of HBD, we constructed a binding model of HBD with heparin octasaccharide (HOS) based on measurements of chemical shift changes and docking studies. Furthermore, the dynamic properties of HBD-HOS and HBD-Trm-HOS complex were assessed by measuring spin relaxation rates. The results showed that the HOS-binding site is composed of two basic clusters consisting of side-chains of residues R13, R14, and K15 and residues K30, R35, and R49. Upon binding HOS, values for the heteronuclear nuclear Overhauser effect near HOS binding sites increased dramatically. CPMG (Carr-Purcell-Meiboom-Gill sequence) experiments as well as R2 relaxation experiment were undertaken to understand millisecond time-scale motions in HBD. There is large relaxation dispersion of residues at Trm and HOS binding sites in free HBD. C-terminal residues such as S34, C48, D51 near the HOS binding sites continued to exhibit slow conformational motions in the HBD-Trm complex while those slow motions disappeared in the bound conformation of HBD with HOS. Collectively, our results demonstrate that the inherent structural flexibilities of the C-terminal region of the HBD are important in the heparin-binding process, and that Trm does not inhibit VEGF-heparin interactions necessary for the biological activities of VEGF.
Related JoVE Video
ER stress-inducible ATF3 suppresses BMP2-induced ALP expression and activation in MC3T3-E1 cells.
Biochem. Biophys. Res. Commun.
PUBLISHED: 11-18-2013
Show Abstract
Hide Abstract
Endoplasmic reticulum (ER) stress suppresses osteoblast differentiation. Activating transcription factor (ATF) 3, a member of the ATF/cAMP response element-binding protein family of transcription factors, is induced by various stimuli including cytokines, hormones, DNA damage, and ER stress. However, the role of ATF3 in osteoblast differentiation has not been elucidated. Treatment with tunicamycin (TM), an ER stress inducer, increased ATF3 expression in the preosteoblast cell line, MC3T3-E1. Overexpression of ATF3 inhibited bone morphogenetic protein 2-stimulated expression and activation of alkaline phosphatase (ALP), an osteogenic marker. In addition, suppression of ALP expression by TM treatment was rescued by silencing of ATF3 using shRNA. Taken together, these data indicate that ATF3 is a novel negative regulator of osteoblast differentiation by specifically suppressing ALP gene expression in preosteoblasts.
Related JoVE Video
Nationwide Surveillance for Pathogenic Microorganisms in Groundwater near Carcass Burials Constructed in South Korea in 2010.
Int J Environ Res Public Health
PUBLISHED: 10-15-2013
Show Abstract
Hide Abstract
Widespread outbreaks of foot-and-mouth disease and avian influenza occurred in South Korea during 2010. In response to the culling of many animals to attenuate the spread of disease, South Korea used mass burial sites to dispose of the large number of carcasses; consequently, concerns about groundwater contamination by leachate from these burial sites are increasing. Groundwater is one of the main sources of drinking water, and its cleanliness is directly related to public health. Thus, this study aimed to evaluate the safety of groundwater around the burial sites (total of 600 sites). A total of 1,200 groundwater samples were collected though the country, and microbial analysis was conducted during two time periods: during the spring (n = 600; April to June 2012) and after rainfall (n = 600; August to October, 2012; fall). Fecal coliform and Escherichia coli were detected in 173 (14.4%) and 85 (7.1%) of the 1,200 samples, respectively. Salmonella spp. and Shigella spp. each were detected only once (0.083%). Clostridium perfringens was detected from 7 groundwater samples (0.583%), and E. coli O157:H7 was not detected. With respect to norovirus, only the GII type was detected from six groundwater samples (0.5%), and enterovirus was detected in 15 groundwater samples (1.25%). The frequency of E. coli that we detected was lower than that found in previous studies conducted in South Korea, but we detected higher frequency of fecal coliform than that observed in a previous report. The contamination frequencies of Salmonella spp. and Shigella spp. were very low, but C. perfringens, which could be an indicator of fecal pollution, was detected in seven regions. Overall, the results of the present study indicate a low possibility of contamination from burial sites. However, consistent monitoring is required to prevent microbial contamination of groundwater near the burial sites.
Related JoVE Video
A cepstral analysis based method for quantifying the depth of anesthesia from human EEG.
Conf Proc IEEE Eng Med Biol Soc
PUBLISHED: 10-11-2013
Show Abstract
Hide Abstract
In this paper, a cepstral analysis based approach to measuring the depth of anesthesia (DoA) is presented. Cepstral analysis is a signal processing technique widely used especially for speech recognition in order to extract speech information regardless of vocal cord characteristics. The resulting index for the DoA is called index based on cepstral analysis (ICep). The Fisher criterion is engaged to evaluate the performance of indices. All analyses are based on a single-channel electroencephalogram (EEG) of 10 human subjects. To validate the proposed technique, ICep is compared with bispectral index (BIS), which is the most commonly used method to estimate the level of consciousness via EEG during general anesthesia. The results show that ICep has high correlation with BIS, and is outstanding in terms of the Fisher criterion and offers faster tracking than BIS in the transition from consciousness to unconsciousness.
Related JoVE Video
A Randomized Comparison of Remifentanil Target-Controlled Infusion Versus Dexmedetomidine Single-Dose Administration: A Better Method for Smooth Recovery From General Sevoflurane Anesthesia.
Am J Ther
PUBLISHED: 10-09-2013
Show Abstract
Hide Abstract
Remifentanil target-controlled infusion and dexmedetomidine single-dose administration are known to reduce airway response and hemodynamic stimulation during anesthetic recovery. We compared the effects of 2 drugs on the prevention of cough during emergence. We enrolled 70 female patients aged 20-60 years with American Society of Anesthesiologists (ASA) I-II who underwent general anesthesia for elective thyroidectomy. The patients were randomly assigned to remifentanil (group R) or dexmedetomidine (group D). Anesthesia was maintained with sevoflurane and effect-site target-controlled infusion of remifentanil. In group D, remifentanil was discontinued, and dexmedetomidine 0.5 ?g/kg was given 10 minutes before the end of surgery. In group R, remifentanil target-controlled infusion at an effective-site concentration of 2.0 ng/mL was maintained during emergence until extubation. The cough grade, hemodynamic values, respiration, and other recovery profiles were evaluated during the periextubation period. The proportion of patients with no cough or just a single cough during extubation was significantly higher in group R than in group D (96.8% vs. 55.9%). The change of mean arterial pressure and heart rate were not significantly different during extubation in both groups. Respiratory rate and the incidence of residual sedation after extubation were lower in group R. There were no desaturation events and no differences in time to extubation or duration of postanesthesia care unit stay in both groups. Remifentanil target-controlled infusion reduces emergence cough from general anesthesia more effectively than single-dose dexmedetomidine. However, a single-dose of dexmedetomidine has the effect with respect to respiratory and hemodynamic stability during emergence.
Related JoVE Video
Accelerated formation of metal oxide thin film at 200 °C using oxygen supplied by a nitric acid additive and residual organic suction vacuum annealing for thin-film transistor applications.
ACS Appl Mater Interfaces
PUBLISHED: 09-04-2013
Show Abstract
Hide Abstract
Oxide semiconductors have gradually replaced amorphous and polycrystalline silicon for thin-film transistor (TFT) because of their high mobility and large-area uniformity. Especially, the oxide semiconductors have also achieved the low-cost manufacturing using a solution process. However, because the solution-processed oxide semiconductors require a high thermal energy to form the oxide thin film, the additional solution synthesis and annealing process are needed for low-temperature solution process. Because the conventional solution-processed oxide thin films have low oxidation level and high residual organic concentration at low annealing temperature, we propose the novel solution process that includes the nitric acid additive and the vacuum ambient annealing as an oxidizing agent and a residual organic suction, respectively. Therefore, we have successfully developed the simple oxide solution process and the soluble InZnO TFT with high field-effect mobility of 3.38 cm(2)/(V s) at 200 °C.
Related JoVE Video
Nucleofection-mediated ?1,3-galactosyltransferase gene inactivation and membrane cofactor protein expression for pig-to-primate xenotransplantation.
Anim. Biotechnol.
PUBLISHED: 08-17-2013
Show Abstract
Hide Abstract
Xenotransplantation of pig organs into primates leads to hyperacute rejection (HAR). Functional ablation of the pig ? 1,3-galactosyltransferase (GalT) gene, which abrogates expression of the Gal ? 1-3Gal ? 1-4GlcNAc-R (Gal) antigen, which inhibits HAR. However, antigens other than Gal may induce immunological rejection by their cognate antibody responses. Ultimately, overexpression of complement regulatory proteins reduces acute humoral rejection by non-Gal antibodies when GalT is ablated. In this study, we developed a vector-based strategy for ablation of GalT function and concurrent expression of membrane cofactor protein (MCP, CD46). We constructed an MCP expression cassette (designated as MCP-IRESneo) and inserted between the left and the right homologous arms to target exon 9 of the GalT gene. Nucleofection of porcine ear skin fibroblasts using the U-023 and V-013 programs resulted in high transfection efficiency and cell survival. We identified 28 clones in which the MCP-IRESneo vector had been successfully targeted to exon 9 of the GalT gene. Two of those clones, with apparent morphologically mitotic fibroblast features were selected through long-term culture. GalT gene expression was downregulated in these 2 clones. Importantly, MCP was shown to be efficiently expressed at the cell surface and to efficiently protect cell lysis against normal human complement serum attack in vitro.
Related JoVE Video
Generation of leukemia inhibitory factor-dependent induced pluripotent stem cells from the massachusetts general hospital miniature pig.
Biomed Res Int
PUBLISHED: 08-08-2013
Show Abstract
Hide Abstract
The generation and application of porcine induced pluripotent stem cells (iPSCs) may enable the testing for safety and efficacy of therapy in the field of human regenerative medicine. Here, the generation of iPSCs from the Massachusetts General Hospital miniature pig (MGH minipig) established for organ transplantation studies is reported. Fibroblasts were isolated from the skin of the ear of a 10-day-old MGH minipig and transduced with a cocktail of six human factors: POU5F1, NANOG, SOX2, C-MYC, KLF4, and LIN28. Two distinct types of iPSCs were generated that were positive for alkaline phosphatase activity, as well as the classical pluripotency markers: Oct4, Nanog, Sox2, and the surface marker Ssea-1. Only one of two porcine iPSC lines differentiated into three germ layers both in vitro and in vivo. Western blot analysis showed that the porcine iPSCs were dependent on LIF or BMP-4 to sustain self-renewal and pluripotency. In conclusion, the results showed that human pluripotent factors could reprogram porcine ear fibroblasts into the pluripotent state. These cells may provide a useful source of cells that could be used for the treatment of degenerative and genetic diseases and agricultural research and application.
Related JoVE Video
Capacitive Epidermal Electronics for Electrically Safe, Long-Term Electrophysiological Measurements.
Adv Healthc Mater
PUBLISHED: 08-07-2013
Show Abstract
Hide Abstract
Integration of capacitive sensing capabilities to epidermal electronic systems (EES) can enhance the robustness in operation for electrophysiological signal measurement. Capacitive EES designs are reusable, electrically safe, and minimally sensitive to motion artifacts. Experiments on human subjects illustrate levels of fidelity in ECG, EMG, and EOG recordings comparable to those of standard gel electrodes and of direct contact EES electrodes.
Related JoVE Video
Two-axis MEMS scanner with transfer-printed high-reflectivity, broadband monolithic silicon photonic crystal mirrors.
Opt Express
PUBLISHED: 06-06-2013
Show Abstract
Hide Abstract
We present a two-axis electrostatic MEMS scanner with high-reflectivity monolithic single-crystal-silicon photonic crystal (PC) mirrors suitable for applications in harsh environments. The reflective surfaces of the MEMS scanner are transfer-printed PC mirrors with low polarization dependence, low angular dependence, and reflectivity over 85% in the wavelength range of 1490nm~1505nm and above 90% over the wavelength band of 1550~1570nm. In static mode, the scanner has total scan range of 10.2° on one rotation axis and 7.8° on the other. Dynamic operation on resonance increase the scan range to 21° at 608Hz around the outer rotation axis and 9.5° at 1.73kHz about the inner rotation axis.
Related JoVE Video
Structure-activity relationships of cecropin-like peptides and their interactions with phospholipid membrane.
BMB Rep
PUBLISHED: 05-29-2013
Show Abstract
Hide Abstract
Cecropin A and papiliocin are novel 37-residue cecropin-like antimicrobial peptides isolated from insect. We have confirmed that papiliocin possess high bacterial cell selectivity and has an ?-helical structure from Lys(3) to Lys(21) and from Ala(25) to Val(35), linked by a hinge region. In this study, we demonstrated that both peptides showed high antimicrobial activities against multi-drug resistant Gram negative bacteria as well as fungi. Interactions between these cecropin-like peptides and phospholipid membrane were studied using CD, dye leakage experiments, and NMR experiments, showing that both peptides have strong permeabilizing activities against bacterial cell membranes and fungal membranes as well as Trp(2) and Phe(5) at the N-terminal helix play an important role in attracting cecropin-like peptides to the negatively charged bacterial cell membrane. Cecropin-like peptides can be potent peptide antibiotics against multi-drug resistant Gram negative bacteria and fungi.
Related JoVE Video
Improvement of Cloning Efficiency in Minipigs Using Post-thawed Donor Cells Treated with Roscovitine.
Mol. Biotechnol.
PUBLISHED: 05-17-2013
Show Abstract
Hide Abstract
Massachusetts General Hospital miniature pigs (MGH minipigs) have been established for organ transplantation studies across the homozygous major histocompatibility complex, but cloning efficiency of MGH minipigs is extremely low. This study was designed to increase the productivity of MGH minipigs by nuclear transfer of post-thaw donor cells after 1 h co-incubation with roscovitine. The MGH minipig cells were genetically modified with GT KO (alpha1,3-galactosyltransferase knock-out) and hCD46 KI (human CD46 knock-in) and used as donor cells. The GT KO/hCD46 KI donor cells were cultured for either 3 days (control group) or 1 h after thawing with 15 ?M roscovitine (experimental group) prior to the nuclear transfer. The relative percentage of the transgenic donor cells that entered into G0/G1 was 93.7 % (±2.54). This was different from the donor cells cultured for 1 h with the roscovitine-treated group (84.6 % ±4.6) (P < 0.05) and without roscovitine (78.6 % ±5.5) (P < 0.01), respectively. The pregnancy rate and delivery rate in the roscovitine group (8/12 and 6/8, respectively) were significantly higher (P < 0.01) than those in the control group (6/19 and 3/6, respectively). In the experimental group, 12 GT KO/hCD46 KI transgenic minipigs were successfully generated, and five minipigs among them survived for more than 6 months so far. The recipient-based individual cloning efficiency ranged from 0.74 to 2.54 %. In conclusion, gene-modified donor cells can be used for cloning of MGH minipigs if the cells are post-thawed and treated with roscovitine for 1 h prior to nuclear transfer.
Related JoVE Video
Materials and optimized designs for human-machine interfaces via epidermal electronics.
Adv. Mater. Weinheim
PUBLISHED: 04-29-2013
Show Abstract
Hide Abstract
Thin, soft, and elastic electronics with physical properties well matched to the epidermis can be conformally and robustly integrated with the skin. Materials and optimized designs for such devices are presented for surface electromyography (sEMG). The findings enable sEMG from wide ranging areas of the body. The measurements have quality sufficient for advanced forms of human-machine interface.
Related JoVE Video
Norovirus contamination levels in ground water treatment systems used for food-catering facilities in South Korea.
Viruses
PUBLISHED: 04-24-2013
Show Abstract
Hide Abstract
This study aimed to inspect norovirus contamination of groundwater treatment systems used in food-catering facilities located in South Korea. A nationwide study was performed in 2010. Water samples were collected and, for the analysis of water quality, the temperature, pH, turbidity, and residual chlorine content were assessed. To detect norovirus genotypes GI and GII, RT-PCR and semi-nested PCR were performed with specific NV-GI and NV-GII primer sets, respectively. The PCR products amplified from the detected strains were then subjected to sequence analyses. Of 1,090 samples collected in 2010, seven (0.64%) were found to be norovirus-positive. Specifically, one norovirus strain was identified to have the GI-6 genotype, and six GII strains had the GII, GII-3, GII-4, and GII-17 genotypes. The very low detection rate of norovirus most likely reflects the preventative measures used. However, this virus can spread rapidly from person to person in crowded, enclosed places such as the schools investigated in this study. To promote better public health and sanitary conditions, it is necessary to periodically monitor noroviruses that frequently cause epidemic food poisoning in South Korea.
Related JoVE Video
Binding model for eriodictyol to Jun-N terminal kinase and its anti-inflammatory signaling pathway.
BMB Rep
PUBLISHED: 04-23-2013
Show Abstract
Hide Abstract
The anti-inflammatory activity of eriodictyol and its mode of action were investigated. Eriodictyol suppressed tumor necrosis factor (mTNF)-?, inducible nitric oxide synthase (miNOS), interleukin (mIL)-6, macrophage inflammatory protein (mMIP)-1, and mMIP-2 cytokine release in LPS-stimulated macrophages. We found that the anti-inflammatory cascade of eriodictyol is mediated through the Toll-like Receptor (TLR)4/CD14, p38 mitogen-activated protein kinases (MAPK), extracellular-signal-regulated kinase (ERK), Jun-N terminal kinase (JNK), and cyclooxygenase (COX)-2 pathway. Fluorescence quenching and saturation-transfer difference (STD) NMR experiments showed that eriodictyol exhibits good binding affinity to JNK, 8.79 × 10(5) M(-1). Based on a docking study, we propose a model of eriodictyol and JNK binding, in which eriodictyol forms 3 hydrogen bonds with the side chains of Lys55, Met111, and Asp169 in JNK, and in which the hydroxyl groups of the B ring play key roles in binding interactions with JNK. Therefore, eriodictyol may be a potent anti-inflammatory inhibitor of JNK. [BMB Reports 2013; 46(12): 594-599].
Related JoVE Video
Onyx Embolization of Dural Arteriovenous Fistula, using Scepter C Balloon Catheter: a Case Report.
Neurointervention
PUBLISHED: 04-20-2013
Show Abstract
Hide Abstract
We report our experience using Onyx for embolization of dural arteriovenous fistula (DAVF) under dual lumen balloon catheter flow arrest. Transfemoral cerebral angiography revealed a superior sagittal sinus (SSS) DAVF that was supplied via multiple branches of the external carotid arteries, the right anterior cerebral arteries, and the meningeal branches of the internal carotid artery. There was no anterograde venous drainage through the SSS, and venous drainage was almost retrograde through the medullary and cortical veins. Under general anesthesia, a transvenous approach was utilized to place the microcatheter close to the fistula site. After intravenous embolization with various coils, DAVF was partially occluded; Balloon catheter gained access to the DAVF via the right middle meningeal artery. We injected Onyx through the Scepter C catheter, after which DAVF was nearly completely occluded. Balloon-assisted Onyx embolization is a feasible and effective approach for the management of DAVF.
Related JoVE Video
Synovial osteochondromatosis in the subacromial bursa mimicking calcific tendinitis: Sonographic diagnosis.
J Clin Ultrasound
PUBLISHED: 04-19-2013
Show Abstract
Hide Abstract
Synovial osteochondromatosis is an idiopathic benign metaplasia of the synovial membrane rarely found in an extra-articular bursa. We describe the case of a 55-year-old woman with synovial osteochondromatosis in the subacromial bursa mimicking calcific tendinitis. Plain radiographs showed a radiopaque mass over the middle facet of the greater tuberosity, suggesting calcific tendinitis. Sonography, however, showed a loose body in the subacromial bursa, and no evidence of calcification inside the rotator cuff. © 2013 Wiley Periodicals, Inc. J Clin Ultrasound, 2013.
Related JoVE Video
Simultaneous detection of waterborne viruses by multiplex real-time PCR.
J. Microbiol.
PUBLISHED: 04-05-2013
Show Abstract
Hide Abstract
Norovirus, Rotavirus group A, the Hepatitis A virus, and Coxsackievirus are all common causes of gastroenteritis. Conventional diagnoses of these causative agents are based on antigen detection and electron microscopy. To improve the diagnostic potential for viral gastroenteritis, internally controlled multiplex real-time polymerase chain reaction (PCR) methods have been recently developed. In this study, individual real-time PCRs were developed and optimized for specific detections of Norovirus genogroup I, Norovirus genogroup II, Rotavirus group A, the Hepatitis A virus, and Coxsackievirus group B1. Subsequently, individual PCRs were combined with multiplex PCR reactions. In general, multiplex real-time PCR assays showed comparable sensitivities and specificities with individual assays. A retrospective clinical evaluation showed increased pathogen detection in 29% of samples using conventional PCR methods. Prospective clinical evaluations were detected in 123 of the 227 (54%) total samples used in the multiplex real-time PCR analysis. The Norovirus genogroup II was found most frequently (23%), followed by Rotavirus (20%), the Hepatitis A virus (4.5%), Coxsackievirus (3.5%), and Norovirus genogroup I (2.6%). Internally controlled multiplex real-time PCR assays for the simultaneous detection of Rotavirus, Coxsackievirus group B, the Hepatitis A virus, and Norovirus genogroups I and II showed significant improvement in the diagnosis of viral gastroenteritis.
Related JoVE Video
Structural and dynamic features of cold-shock proteins of Listeria monocytogenes, a psychrophilic bacterium.
Biochemistry
PUBLISHED: 04-01-2013
Show Abstract
Hide Abstract
Cold-shock proteins (Csps), proteins expressed when the ambient temperature drops below the growth-supporting temperature, bind to single-stranded nucleic acids and act as RNA chaperones to regulate translation. Listeria monocytogenes is a psychrophilic food-borne pathogen that is problematic for the food industry. Structures of Csps from psychrophilic bacteria have not yet been studied. Despite dramatic differences in the thermostability of Csps of various thermophilic microorganisms, these proteins share a high degree of primary sequence homology and a high degree of three-dimensional structural similarity. Here, we investigated the structural and dynamic features as well as the thermostability of L. monocytogenes CspA (Lm-CspA). Lm-CspA has a five-stranded ?-barrel structure with hydrophobic core packing and two salt bridges. When heptathymidine (dT(7)) binds, values for the heteronuclear nuclear Overhauser effect and order parameters of residues in surface loop regions near nucleic acid binding sites increase dramatically. Moreover, Carr-Purcell-Meiboom-Gill experiments showed that slow motions observed for the nucleic acid binding residues K7, W8, F15, F27, and R56 disappeared in Lm-CspA-dT(7). Lm-CspA is less thermostable than mesophilic and thermophilic Csps, with a lower melting temperature (40 °C). The structural flexibility that accompanies longer surface loops and less hydrophobic core packing and a number of salt bridges and unfavorable electrostatic repulsion are likely key factors in the low thermostability of Lm-CspA. This implies that the large conformational flexibility of psychrophilic Lm-CspA, which more easily accommodates nucleic acids at low temperature, is required for RNA chaperone function under cold-shock conditions and for the cold adaptation of L. monocytogenes.
Related JoVE Video
Mycobacterium intracellulare Pulmonary Disease with Endobronchial Caseation in a Patient Treated with Methotrexate.
Tuberc Respir Dis (Seoul)
PUBLISHED: 03-07-2013
Show Abstract
Hide Abstract
Methotrexate (MTX) has been established as a standard disease-modifying anti-rheumatic drug. If adequate disease control is achieved for a reasonable period of time, tapering the MTX dosage is recommended because the chronic use of MTX can result in opportunistic infection. We present here a case of a woman with rheumatoid arthritis taking MTX, and the woman developed actively caseating endobronchial Mycobacterium intracellulare disease with pulmonary infiltrations. After discontinuing the MTX, the patient was able to tolerate 18 months of antimycobacterial treatment without flare ups of rheumatoid arthritis, and she completely recovered from nontuberculous mycobacterial respiratory disease.
Related JoVE Video
Backbone dynamics of an atypical orphan response regulator protein, Helicobacter pylori 1043.
Mol. Cells
PUBLISHED: 02-21-2013
Show Abstract
Hide Abstract
An atypical orphan response regulator protein, HP1043 (HP-RR) in Helicobacter pylori, is proven to be essential for cell growth and does not require the well known phosphorelay scheme. HP-RR was identified as a symmetric dimer with two functional domains, an N-terminal regulatory domain (HP-RR(r)) and a C-terminal effector domain (HP-RR(e)). HP-RR is a new class of response regulator, as a phosphorylation-independent regulator. Previously, we have presented a detailed three-dimensional structure of HP-RR using NMR spectroscopy and X-ray crystallography. In this study, in order to understand the functional importance of flexibilities in HP-RR(r) and HP-RR(e), T1, T2, heteronuclear NOE experiments have been performed and backbone dynamics of HP-RR(r) and HP-RR(e) were investigated. HP-RR(r) is a symmetric dimer and the interface region, ?4-?5-?5 of dimer, showed high rigidity (high S (2) values). Site of rearrangements associated with phosphorylation of HP-RR(r) (Ser(75): R ex = 3.382, Ile(95): R ex = 5.228) showed slow chemical exchanges. HP-RR(e) is composed of three ?-helices flanked on two sides by anti-parallel ?-sheets. Low order parameters as well as conformational exchanges in the centers of loop regions known as the DNA binding site and transcription site of HP-RR(e) suggested that flexibility of HP-RR(e) is essential for interaction with DNA. In conclusion, backbone dynamics information for HP-RR implies that structural flexibilities in HP-RR(r) are necessary for the phosphorylation site and the dynamic nature of HP-RR(e) is essential for the regulation of interaction between protein and DNA.
Related JoVE Video
The phosphodiesterase 4 inhibitor rolipram protects against cigarette smoke extract-induced apoptosis in human lung fibroblasts.
Eur. J. Pharmacol.
PUBLISHED: 02-14-2013
Show Abstract
Hide Abstract
Cigarette smoke, a major causative agent of chronic obstructive pulmonary disease (COPD), induces lung cell death by incompletely understood mechanisms. The induction of apoptosis in lung structural cells by cigarette smoke may contribute to the pathogenesis of emphysema. Phosphodiesterase-4 (PDE4) inhibitors are anti-inflammatory agents used in COPD therapy that can prevent cigarette smoke-induced emphysema in mice. We investigated the effect of rolipram, a first generation PDE4 inhibitor, on the regulation of cigarette smoke-induced apoptosis. Human lung fibroblast (MRC-5) cells were exposed to cigarette smoke extract (CSE). Cell viability and apoptosis were determined by MTT assay and Annexin-V staining, respectively. Caspase activation was determined by Western immunoblot analysis. Rolipram protected against cell death and increased viability in MRC-5 fibroblasts after CSE exposure. Furthermore, rolipram protected against apoptosis, decreased caspase-3 and -8 cleavage in MRC-5 cells exposed to CSE. Pre-treatment with rolipram enhanced Akt phosphorylation and associated cytoprotection in CSE-treated cells, which could be reversed by the PI3K inhibitor LY294002 partly. In conclusion, rolipram protects against apoptosis of MRC-5 cells through inhibition of caspase-3 and caspase-8. Rolipram may represent an effective therapeutic agent to reduce cigarette smoke-induced apoptosis of lung fibroblasts.
Related JoVE Video
Coiling as Retreatment in Intracranial Aneurysm of de novo Formation or Regrowth: Case Report.
Neurointervention
PUBLISHED: 02-05-2013
Show Abstract
Hide Abstract
Development of de novo aneurysm or aneurysm regrowth after complete clipping of an intracranial aneurysm is rare. We report coiled cases of de novo aneurysm and aneurysm regrowth. We retrospectively reviewed 107 cases of intracranial aneurysm coiling performed in our hospital, identifying five cases of coiled aneurysm that were de novo aneurysm or aneurysm regrowth. In all the cases, total or near total occlusion was seen. There were no complications related to the procedure. In two of the three patients with ruptured aneurysms, consciousness level on admission was stupor. When the patient was discharged after the treatment, one of them had 4 of the modified Rankin Scale. The other one had 5 on discharge. The rest three patients had 0. As for a de novo aneurysm or a regrowth of aneurysm, coiling may be considered when clipping is difficult.
Related JoVE Video
Postural stability in patients with anterior cruciate ligament tears with and without medial meniscus tears.
Knee Surg Sports Traumatol Arthrosc
PUBLISHED: 01-26-2013
Show Abstract
Hide Abstract
To compare postural stability in patients with isolated anterior cruciate ligament (ACL) tears and ACL tears with associated meniscal tears.
Related JoVE Video
MiR-433 mediates ERR?-suppressed osteoblast differentiation via direct targeting to Runx2 mRNA in C3H10T1/2 cells.
Life Sci.
PUBLISHED: 01-06-2013
Show Abstract
Hide Abstract
MicroRNAs (miRNA) are involved in various biological processes including cellular differentiation. However, the role of miR-433 in osteoblast differentiation remains poorly understood. The objective of this study was to investigate the effect of miR-433 on BMP2-induced osteoblast differentiation.
Related JoVE Video
De novo design and synthesis of ultra-short peptidomimetic antibiotics having dual antimicrobial and anti-inflammatory activities.
PLoS ONE
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
Much attention has been focused on the design and synthesis of potent, cationic antimicrobial peptides (AMPs) that possess both antimicrobial and anti-inflammatory activities. However, their development into therapeutic agents has been limited mainly due to their large size (12 to 50 residues in length) and poor protease stability.
Related JoVE Video
Regulatory roles of ganglioside GQ1b in neuronal cell differentiation of mouse embryonic stem cells.
BMB Rep
PUBLISHED: 12-23-2011
Show Abstract
Hide Abstract
Gangliosides play an important role in neuronal differentiation processes. The regulation of ganglioside levels is related to the induction of neuronal cell differentiation. In this study, the ST8Sia5 gene was transfected into mESCs and then differentiated into neuronal cells. Interestingly, ST8Sia5 gene transfected mESCs expressed GQ1b by HPTLC and immunofluorescence analysis. To investigate the effects of GQ1b over-expression in neurogenesis, neuronal cells were differentiated from GQ1b expressing mESCs in the presence of retinoic acid. In GQ1b expressing mESCs, increased EBs formation was observed. After 4 days, EBs were co-localized with GQ1b and nestin, and GFAP. Moreover, GQ1b co-localized with MAP-2 expressing cells in GQ1b expressing mESCs in 7-day-old EBs. Furthermore, GQ1b expressing mESCs increased the ERK1/2 MAP kinase pathway. These results suggest that the ST8Sia5 gene increases ganglioside GQ1b and improves neuronal differentiation via the ERK1/2 MAP kinase pathway.
Related JoVE Video
Multiple compartment syndrome in a pediatric patient with CML.
J Pediatr Orthop
PUBLISHED: 11-22-2011
Show Abstract
Hide Abstract
Compartment syndrome is a limb-threatening and life-threatening emergency resulting from elevated intracompartmental pressure. Prompt surgical intervention and treatment are necessary to prevent irreparable damage to muscle and nerve tissues. Leukemic infiltration of the muscle is an unusual cause of compartment syndrome and has been documented to occur secondary to hyperleukocytic leukemias, most commonly in acute myeloid leukemia. We present a rare case of multiple compartment syndrome in the buttock and thigh of an 11-year-old male patient with chronic myelomonocytic leukemia. The diagnosis of acute compartment syndrome was delayed, causing irreversible tissue damage. Physicians are generally unfamiliar with leukemia-induced complications and may not initially suspect leukemic compartment syndrome because of its rarity. Awareness of its clinical features is critical, because early diagnosis and prompt surgical debridement can prevent significant morbidity and even death.
Related JoVE Video
Structure and function of papiliocin with antimicrobial and anti-inflammatory activities isolated from the swallowtail butterfly, Papilio xuthus.
J. Biol. Chem.
PUBLISHED: 09-29-2011
Show Abstract
Hide Abstract
Papiliocin is a novel 37-residue cecropin-like peptide isolated recently from the swallowtail butterfly, Papilio xuthus. With the aim of identifying a potent antimicrobial peptide, we tested papiliocin in a variety of biological and biophysical assays, demonstrating that the peptide possesses very low cytotoxicity against mammalian cells and high bacterial cell selectivity, particularly against Gram-negative bacteria as well as high anti-inflammatory activity. Using LPS-stimulated macrophage RAW264.7 cells, we found that papiliocin exerted its anti-inflammatory activities by inhibiting nitric oxide (NO) production and secretion of tumor necrosis factor (TNF)-? and macrophage inflammatory protein (MIP)-2, producing effects comparable with those of the antimicrobial peptide LL-37. We also showed that the innate defense response mechanisms engaged by papiliocin involve Toll-like receptor pathways that culminate in the nuclear translocation of NF-?B. Fluorescent dye leakage experiments showed that papiliocin targets the bacterial cell membrane. To understand structure-activity relationships, we determined the three-dimensional structure of papiliocin in 300 mm dodecylphosphocholine micelles by NMR spectroscopy, showing that papiliocin has an ?-helical structure from Lys(3) to Lys(21) and from Ala(25) to Val(36), linked by a hinge region. Interactions between the papiliocin and LPS studied using tryptophan blue-shift data, and saturation transfer difference-NMR experiments revealed that Trp(2) and Phe(5) at the N-terminal helix play an important role in attracting papiliocin to the cell membrane of Gram-negative bacteria. In conclusion, we have demonstrated that papiliocin is a potent peptide antibiotic with both anti-inflammatory and antibacterial activities, and we have laid the groundwork for future studies of its mechanism of action.
Related JoVE Video
Biochemical characterization and FAD-binding analysis of oleate hydratase from Macrococcus caseolyticus.
Biochimie
PUBLISHED: 09-13-2011
Show Abstract
Hide Abstract
A putative fatty acid hydratase gene from Macrococcus caseolyticus was cloned and expressed in Escherichia coli. The recombinant enzyme was a 68 kDa dimer with a molecular mass of 136 kDa. The enzymatic products formed from fatty acid substrates by the putative enzyme were isolated with high purity (>99%) by solvent fractional crystallization at low temperature. After the identification by GC-MS, the purified hydroxy fatty acids were used as standards to quantitatively determine specific activities and kinetic parameters for fatty acids as substrates. Among the fatty acids evaluated, specific activity and catalytic efficiency (k(cat)/K(m)) were highest for oleic acid, indicating that the putative fatty acid hydratase was an oleate hydratase. Hydration occurred only for cis-9-double and cis-12-double bonds of unsaturated fatty acids without any trans-configurations. The maximum activity for oleate hydration was observed at pH 6.5 and 25 °C with 2% (v/v) ethanol and 0.2 mM FAD. Without FAD, all catalytic activity was abolished. Thus, the oleate hydratase is an FAD-dependent enzyme. The residues G29, G31, S34, E50, and E56, which are conserved in the FAD-binding motif of fatty acid hydratases (GXGXXG((A/S))X((15-21))E((D))), were selected by alignment, and the spectral properties and kinetic parameters of their alanine-substituted variants were analyzed. Among the five variants, G29A, G31A, and E56A showed no interaction with FAD and exhibited no activity. These results indicate that G29, G31, and E56 are essential for FAD-binding.
Related JoVE Video
Monitoring the depth of anesthesia from rat EEG using modified Shannon entropy analysis.
Conf Proc IEEE Eng Med Biol Soc
PUBLISHED: 08-29-2011
Show Abstract
Hide Abstract
In this paper, an entropy based method for quantifying the depth of anesthesia from rat EEG is presented. The proposed index for the depth of anesthesia called modified Shannon entropy (MShEn) is based on Shannon entropy (ShEn) and spectral entropy (SpEn) which are widely used for analyzing non-stationary signals. Discrimination power (DP), as a performance indicator for indexes, is defined and used to derive the final index for the depth of anesthesia. For experiment, EEG from anesthetized rats are measured and analyzed by using MShEn. MShEn shows both high stability and high correlation with other indexes for depth of anesthesia.
Related JoVE Video
Virtual Bernese osteotomy using three-dimensional computed tomography in hip dysplasia.
Arch Orthop Trauma Surg
PUBLISHED: 07-20-2011
Show Abstract
Hide Abstract
Accurate assessment of acetabular morphology and its relationship to the femoral head is essential for planning a periacetabular osteotomy. We observed the acetabular coverage after virtual Bernese osteotomy using computer-aided technique.
Related JoVE Video
Anterior cruciate ligament rupture in gouty arthritis.
Knee Surg Sports Traumatol Arthrosc
PUBLISHED: 07-18-2011
Show Abstract
Hide Abstract
A 34-year-old male presented with right knee instability without any trauma. He had been diagnosed with right knee gouty arthritis 2 years prior. An arthroscopic examination revealed abundant calcific material deposited around the knee joint, including in the ACL tissue, and that the ACL was torn at the femoral attachment site. Treatment involved a synovectomy to remove calcific material, followed by an ACL reconstruction. Histology evaluation revealed gouty arthritis with the presence of tophi in the synovium, soft tissue, and ACL tissue. The case presented here indicates the possibility of pathologic rupture of the ACL associated with gouty tophus infiltration of that ligament. Level of evidence IV.
Related JoVE Video
Discovery of novel selective inhibitors of Staphylococcus aureus ?-ketoacyl acyl carrier protein synthase III.
Eur J Med Chem
PUBLISHED: 06-29-2011
Show Abstract
Hide Abstract
?-Ketoacyl-acyl carrier protein synthase III (KAS III) is a condensing enzyme in bacterial fatty acid synthesis and a potential target while designing novel antibiotics. In our previous report, we discovered the lead compound YKAs3003, which serves as an inhibitor of Escherichia coli KAS III (ecKAS III), and determined a reliable pharmacophore map from in silico screening. In this study, we determined two pharmacophore maps from receptor-oriented pharmacophore-based in silico screening of the x-ray structure of Staphylococcus aureus KAS III (saKAS III) to identify potent saKAS III inhibitors. We discovered a new potential inhibitor (6) with broad-spectrum antimicrobial activity and 0.8 nM binding affinity for saKAS III, proving the reliability of our pharmacophore map. Using optimization procedures, we identified three new antimicrobial saKAS III inhibitors: 6c (2,4-dichloro-benzoic acid (2,3,4-trihydroxy-benzylidene)-hydrazide), 6e (4-[(3-chloro-pyrazin-2-yl)-hydrazonomethyl]-benzene-1,3-diol), and 6 (4-[(5-trifluoromethyl-pyridin-2-yl)-hydrazonomethyl]-benzene-1,3-diol). All three inhibitors have a novel 4-hydrazonomethyl-benzene-1,3-diol core structure. These inhibitors exhibited high binding affinity to saKAS III and highly selective antimicrobial activities against S. aureus and methicillin-resistant S. aureus, with minimal inhibitory concentration values of 1-2 ?g/mL.
Related JoVE Video
Transvenous embolization of cavernous sinus dural arteriovenous fistula using the direct superior ophthalmic vein approach: a case report.
Neurointervention
PUBLISHED: 05-27-2011
Show Abstract
Hide Abstract
Transvenous coil embolization has been successfully applied for the treatment of cavernous sinus dural arteriovenous fistula (CSDAVF). Unfortunately, the technique cannot be applied in cases of poor or absent inferior petrosal sinus or facial venous access route to the fistula. Recently, we experienced a successful embolization using direct superior ophthalmic vein approach in cases of CSDAVF which were no opacification of inferior petrosal sinus or facial vein.
Related JoVE Video
Use of organic acids to inactivate Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on organic fresh apples and lettuce.
J. Food Sci.
PUBLISHED: 05-27-2011
Show Abstract
Hide Abstract
This study was undertaken to investigate the antimicrobial effect of organic acids against?Escherichia coli?O157:H7,?Salmonella?Typhimurium, and?Listeria monocytogenes?on whole red organic apples and lettuce. Several studies have been conducted to evaluate organic acids as sanitizers. However, no studies have compared antimicrobial effects of various organic acids on organic fresh produce, including evaluation of color changes of produce. Apples and lettuce were inoculated with a cocktail of 3 strains each of 3 foodborne pathogens provided above and treated with 1% and 2% organic acids (propionic, acetic, lactic, malic, and citric acid) for 0, 0.5, 1, 5, and 10 min. With increasing treatment time and acid concentration, organic acid treatments showed significant reduction compared to the control treatment (distilled water), and differences in antimicrobial effects between organic acids were observed. After 10 min of treatment with 1% and 2% organic acids in apples, propionic (0.92 to 2.75 log reduction), acetic (0.52 to 2.78 log reduction), lactic (1.69 to >3.42 log reduction), malic (1.48 to >3.42 log reduction), and citric acid (1.52 to >3.42 log reduction) exhibited significant (P?< 0.05) antibacterial effects against 3 foodborne pathogens compared to the control treatment. In lettuce, propionic (0.93 to 1.52 log reduction), acetic (1.13 to 1.74 log reduction), lactic (1.87 to 2.54 log reduction), malic (2.32 to 2.98 log reduction), and citric acid (1.85 to 2.86 log reduction) showed significant (P?< 0.05) effects compared to the control treatment. Changes in sample color subjected to organic acids treatment were not significant during storage.
Related JoVE Video
Molecular characterization of a novel thermostable mannose-6-phosphate isomerase from Thermus thermophilus.
Biochimie
PUBLISHED: 05-24-2011
Show Abstract
Hide Abstract
Mannose-6-phosphate isomerase catalyzes the interconversion of mannose-6-phosphate and fructose-6-phosphate. The gene encoding a putative mannose-6-phosphate isomerase from Thermus thermophilus was cloned and expressed in Escherichia coli. The native enzyme was a 29 kDa monomer with activity maxima for mannose 6-phosphate at pH 7.0 and 80 °C in the presence of 0.5 mM Zn(2+) that was present at one molecule per monomer. The half-lives of the enzyme at 65, 70, 75, 80, and 85 °C were 13, 6.5, 3.7, 1.8, and 0.2 h, respectively. The 15 putative active-site residues within 4.5 Å of the substrate mannose 6-phosphate in the homology model were individually replaced with other amino acids. The sequence alignments, activities, and kinetic analyses of the wild-type and mutant enzymes with amino acid changes at His50, Glu67, His122, and Glu132 as well as homology modeling suggested that these four residues are metal-binding residues and may be indirectly involved in catalysis. In the model, Arg11, Lys37, Gln48, Lys65 and Arg142 were located within 3 Å of the bound mannose 6-phosphate. Alanine substitutions of Gln48 as well as Arg142 resulted in increase of K(m) and dramatic decrease of k(cat), and alanine substitutions of Arg11, Lys37, and Lys65 affected enzyme activity. These results suggest that these 5 residues are substrate-binding residues. Although Trp13 was located more than 3 Å from the substrate and may not interact directly with substrate or metal, the ring of Trp13 was essential for enzyme activity.
Related JoVE Video
Bilateral sacroiliac joint dislocation (anterior and posterior) with triradiate cartilage injury: a case report.
J Orthop Trauma
PUBLISHED: 05-18-2011
Show Abstract
Hide Abstract
Pediatric sacroiliac joint injuries are uncommon. Significant pelvis ring disruptions in children are rare, and their management is complicated by patient size, differences in bony architecture, and future growth and remodeling potential. We present a rare case of anterior sacroiliac joint dislocation associated with triradiate cartilage injury with a posterior sacroiliac dislocation on the contralateral side. This appears to be the first such case reported in the literature.
Related JoVE Video
Dynamics of a heparin-binding domain of VEGF(165) complexed with its inhibitor triamterene.
Biochemistry
PUBLISHED: 05-13-2011
Show Abstract
Hide Abstract
Vascular endothelial growth factor (VEGF), which has neurotrophic and neuroprotective effects in addition to its major role in angiogenesis, interacts with A? and accumulates in the senile plaques of Alzheimers disease (AD) patients brains. It is known that A? binds to the heparin-binding domain (HBD) of the 165-amino acid VEGF variant, VEGF(165). In this study, we showed that triamterene (Trm) inhibits VEGF--A? interaction without affecting other biological activities of VEGF or A?. We investigated the importance of structural and dynamic features of HBD for its molecular-recognition processes. The binding model of HBD and Trm was constructed based on measurements of chemical shift changes and docking study. The results showed that the loop region (S11-L17) and F18 at the beginning of the first ?-sheet in the HBD constitute the inhibitor binding site. The N1 atom of pteridine ring of Trm forms hydrogen bonding with backbone amide proton of R13, and the phenyl ring took part in a hydrophobic interaction with the aromatic ring of F18. To investigate the functional importance of the inherent structural flexibility of the HBD in VEGF, the dynamic properties of free HBD and HBD--Trm complex were assessed by measuring spin relaxation rates, and the backbone dynamics were investigated by model-free analysis. The residues in the disordered loop region of the N-terminus exhibited conformational exchanges in free HBD, and flexibility of this loop region decreased dramatically upon binding to Trm, suggesting that A? as well as inhibitor may recognize these unique dynamic features of the HBD. Furthermore, C-terminal residues continued to exhibit slow conformational motions, even in the HBD--Trm complex, implying that these motions at the C-terminus of the HBD might be important for interactions with heparin molecules. The flexibility of HBD demonstrated here should be essential for VEGF function and interaction with other protein partners.
Related JoVE Video
Elbow arthrography in children with an ulnar fracture and occult subluxation of the radial head.
J Pediatr Orthop B
PUBLISHED: 05-10-2011
Show Abstract
Hide Abstract
The purpose of this study was to investigate the use of elbow arthrography for detecting an occult subluxation of the radial head and for verifying the quality of reduction in children with ulnar fractures. The study involved 15 children who underwent closed reduction (with or without percutaneous pinning) based on arthrography for minimally displaced fractures with or without radial head subluxation. The mean age of the patient at the time of surgery was 6 years (range, 3-12 years). Initial diagnoses based on conventional radiographs were compared with arthrographically confirmed final diagnoses in groups of children with ulnar fractures of the olecranon, proximal, and diaphysis. Arthrography confirmed initial radiographic diagnoses in 10 and altered diagnoses in five patients. We conclude that arthrography can be useful for detecting an occult subluxation of the radial head and for evaluating adequate reduction in children of less than 6 years with ulnar fractures.
Related JoVE Video
An E3 ligase complex regulates SET-domain polycomb group protein activity in Arabidopsis thaliana.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 04-25-2011
Show Abstract
Hide Abstract
Transcriptional repression via methylation of histone H3 lysine 27 (H3K27) by the polycomb repressive complex 2 (PRC2) is conserved in higher eukaryotes. The Arabidopsis PRC2 controls homeotic gene expression, flowering time, and gene imprinting. Although downstream target genes and the regulatory mechanism of PRC2 are well understood, much less is known about the significance of posttranslational regulation of PRC2 protein activity. Here, we show the posttranslational regulation of CURLY LEAF (CLF) SET-domain polycomb group (PcG) protein by the F-box protein, UPWARD CURLY LEAF1 (UCL1). Overexpression of UCL1 generates mutant phenotypes similar to those observed in plants with a loss-of-function mutation in the CLF gene. Leaf curling and early flowering phenotypes of UCL1 overexpression mutants, like clf mutants, are rescued by mutations in the AGAMOUS and FLOWERING LOCUS T genes, which is consistent with UCL1 and CLF functioning in the same genetic pathway. Overexpression of UCL1 reduces the level of CLF protein and alters expression and H3K27 methylation of CLF-target genes in transgenic plants, suggesting that UCL1 negatively regulates CLF. Interaction of UCL1 with CLF was detected in plant nuclei and in the yeast two-hybrid system. The UCL1 F-box binds in vivo to components of the E3 ligase complex, which ubiquitylate proteins that are subsequently degraded via the ubiquitin-26S proteasome pathway. Taken together, these results demonstrate the posttranslational regulation of the CLF SET-domain PcG activity by the UCL1 F-box protein in the E3 ligase complex.
Related JoVE Video
Inhibition of ganglioside GD1a synthesis suppresses the differentiation of human mesenchymal stem cells into osteoblasts.
Dev. Growth Differ.
PUBLISHED: 04-16-2011
Show Abstract
Hide Abstract
In this study, we investigated the regulatory role of ganglioside GD1a in the differentiation of osteoblasts from human mesenchymal stem cells (hMSCs) by using lentivirus-containing short hairpin (sh)RNA to knockdown ST3 ?-galactoside ?-2, 3-sialyltransferase 2 (ST3Gal II) mRNA expression. After hMSCs were infected for 72 h with the lentivirus constructed with ST3Gal II shRNAs, the puromycin-resistant cells were selected and subcultured to produce hMSCs with ST3Gal II mRNA knockdown. The hMSCs established from human dental papilla abundantly expressed CD44 and CD105, but not CD45 and CD117. Osteoblasts that differentiated from normal hMSCs showed a significant increase in alkaline phosphatase (ALP) activity and ganglioside GD1a expression level compared with those in hMSCs. Lentiviral infection of hMSCs successfully induced a marked inhibition of ST3Gal II mRNA expression and caused a significant decrease in ALP activity and ganglioside GD1a expression. During osteoblastic differentiation, the increased ALP activity remarkably reduced by suppression of ganglioside GD1a expression by ST3Gal II shRNA. Ganglioside GD1a and ALP were mainly expressed in the cell body of hMSCs and osteoblasts with colocalization. The phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 mitogen-activated protein (MAP) kinase and epidermal growth factor receptor (EGFR) was significantly reduced in the osteoblasts that had differentiated from the hMSCs with ST3Gal II mRNA knockdown. These results suggest that ganglioside GD1a plays an important role in the regulation of osteoblastic differentiation of hMSCs through the activation of ERK 1/2 MAP kinase and EGFR.
Related JoVE Video
Full sequence analysis and characterization of the South Korean Norovirus GII-4 variant CUK-3.
Virol. J.
PUBLISHED: 04-14-2011
Show Abstract
Hide Abstract
Many of researchers have focused on the emerging pathogen, Norovirus, since its first identification as the causing agent of nonbacterial acute gastroenteritis in humans. One of the virulence factors of norovirus, the great genetic diversity attributed to point mutations and recombinations, has brought forth the result of significant changes in the circulating norovirus genotype patterns.
Related JoVE Video
A concise synthesis and antimicrobial activities of 3-polyamino-23,24-bisnorcholanes as steroid-polyamine conjugates.
Bioorg. Med. Chem. Lett.
PUBLISHED: 04-08-2011
Show Abstract
Hide Abstract
A series of steroid-polyamine conjugates were synthesized and evaluated for their antimicrobial activity. This study was focused on the effect of stereochemistry at the C-3 and C-5 of steroids and types of polyamine at C-3 on activity against various human pathogens. All the conjugates exhibited strong antimicrobial activities against Gram-positive strains. Compound 18 was found to be the most potent in these series with a MIC value as low as 1 ?g/mL against the bacterium Staphylococcus aureus ATCC6538P.
Related JoVE Video
A paragonimiasis patient with allergic reaction to praziquantel and resistance to triclabendazole: successful treatment after desensitization to praziquantel.
Korean J. Parasitol.
PUBLISHED: 03-18-2011
Show Abstract
Hide Abstract
Paragonimiasis is an infectious disease caused by trematodes of the genus Paragonimus. This trematode can be treated successfully with praziquantel in more than 90% of the cases. Although praziquantel is generally well tolerated, anaphylactic reactions to this drug have been reported in a few cases. We report here a 46-year-old Korean female with paragonimiasis, presumed to be due to Paragonimus westermani, who displayed an allergic reaction to praziquantel and resistance to triclabendazole treatment. The patient was successfully treated with praziquantel following a rapid desensitization procedure. Desensitization to praziquantel could be considered when no alternative drugs are available.
Related JoVE Video

What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

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