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.
Importance of the Time Interval between Bowel Preparation and Colonoscopy in Determining the Quality of Bowel Preparation for Full-Dose Polyethylene Glycol Preparation.
Gut Liver
PUBLISHED: 11-15-2014
Show Abstract
Hide Abstract
The quality of bowel preparation (QBP) is the important factor in performing a successful colonoscopy. Several factors influencing QBP have been reported; however, some factors, such as the optimal preparation-to-colonoscopy time interval, remain controversial. This study aimed to determine the factors influencing QBP and the optimal time interval for full-dose polyethylene glycol (PEG) preparation.
Related JoVE Video
Intermittent Antibiotic Therapy for Nodular Bronchiectatic Mycobacterium avium Complex Lung Disease.
Am. J. Respir. Crit. Care Med.
PUBLISHED: 11-14-2014
Show Abstract
Hide Abstract
Rationale: Although intermittent, three-times-weekly therapy is recommended for the initial treatment of non-cavitary nodular bronchiectatic Mycobacterium avium complex (MAC) lung disease, supporting data are limited. Objectives: To evaluate the clinical efficacy of intermittent therapy compared with daily therapy for nodular bronchiectatic MAC lung disease. Methods: A retrospective cohort study of 217 patients with treatment-naïve non-cavitary nodular bronchiectatic MAC lung disease. All patients received either daily (n = 99) or intermittent therapy (n = 118) that included clarithromycin or azithromycin, rifampin, and ethambutol. Measurements and Results: Modification of the initial antibiotic therapy occurred more frequently in the daily therapy group than in the intermittent therapy group (46% vs. 21%, P < 0.001); in particular, ethambutol was more frequently discontinued in the daily therapy group than in the intermittent therapy group (24% vs. 1%, P < 0.001). However, the rates of symptomatic improvement, radiographic improvement, and sputum culture conversion were not different between the two groups (daily therapy vs. intermittent therapy; 75% vs. 82%, P = 0.181; 68% vs. 73%, P = 0.402; 76% vs. 67%, P = 0.154; respectively). In addition, the adjusted proportion of sputum culture conversion was similar between the daily therapy (71.3%, 95% confidence interval [CI] 59.1-81.1%) and the intermittent therapy groups (73.6%, 95% CI 62.9-82.2%, P = 0.785). Conclusions: These results suggest that intermittent three-times-weekly therapy with a macrolide, rifampin, and ethambutol is a reasonable initial treatment regimen for patients with non-cavitary nodular bronchiectatic MAC lung disease. Clinical trial registration can be found at www. clinicaltrials.gov, ID NCT00970801.
Related JoVE Video
Anti-inflammatory activity of SMP30 modulates NF-?B through protein tyrosine kinase/phosphatase balance.
J. Mol. Med.
PUBLISHED: 10-23-2014
Show Abstract
Hide Abstract
Recent studies on senescence marker protein-30 (SMP30) have shown that it has an important functional role in the aging process, but its precise participation in cellular works has not been fully determined. We hypothesize that SMP30 plays crucial roles in signaling processes by modulating the balance of protein tyrosine kinase (PTK)/protein tyrosine phosphatase (PTP) and in activating proinflammatory NF-?B. An experimental paradigm of gain and loss of SMP30 function was established using SMP30-overexpressed YPEN-1 cells (herein referred to as "SMP30(+) cells") and SMP30 (Y/-) knockout mouse kidneys. The resulting data show that SMP30 expression suppressed oxidative stress-induced PTK/PTP dysregulation and PP1/2A inactivation in SMP30(+) cells, leading to the suppression of NF-?B activation. In the kidneys of SMP30 (Y/-) mice, SMP30 deficiency was found to induce NF-?B activation via the upstream signaling of NIK/IKK and MAPKs and to upregulate downstream NF-?B-responsive gene expression. In this study, we also demonstrate for the first time that SMP30 deficiency induced PTK activity in SMP30 (Y/-) kidneys, thereby significantly increasing the tyrosine phosphorylation of a catalytic subunit of PP2A (PP2Ac-Tyr307). Based on these findings, we propose that SMP30 involves NF-?B regulation through the PTK/PTP balance and that the age-related decrease of SMP30 causes NF-?B activation, which contributes to an exacerbation of the inflammatory process during aging.
Related JoVE Video
Enhanced current transport and injection in thin-film gallium-nitride light-emitting diodes by laser-based doping.
ACS Appl Mater Interfaces
PUBLISHED: 09-25-2014
Show Abstract
Hide Abstract
This paper reports improvements in the electrical and optical properties of blue-emission gallium nitride (GaN)-based thin-film light-emitting diodes (TFLEDs) after laser-based Si doping (LBSD) of a nitrogen-face n-GaN (denoted as hereafter n-GaN) layer. Experimental results show that the light-output powers of the flat- and rough-surface TFLEDs after LBSD are 52.1 and 11.35% higher than those before LBSD, respectively, at a current of 350 mA, while the corresponding operating voltages are decreased by 0.22 and 0.28 V for the flat- and rough-surface TFLEDs after LBSD, respectively. The reduced operating voltage after LBSD of the top n-GaN layer may result from the remarkably decreased specific contact resistance at the metal/n-GaN interface and the low series resistance of the TFLED device. The LBSD of n-GaN increases the number of nitrogen vacancies, and Si substitutes for Ga (SiGa) at the metal/n-GaN interface to produce highly Si-doped regions in n-GaN, leading to a decrease in the Schottky barrier height and width. As a result, the specific contact resistances are significantly decreased to 1.56 × 10(-5) and 2.86 × 10(-5) ? cm(2) for the flat- and rough-surface samples after LBSD, respectively. On the other hand, the increased light-output power after LBSD can be explained by the uniform current spreading, efficient current injection, and enhanced light scattering resulting from the low contact resistivity, low lateral current resistance, and additional textured surface, respectively. Furthermore, LBSD did not degrade the electrical properties of the TFLEDs owing to low reverse leakage currents. The results indicate that our approach could potentially enable high-efficiency and high-power capabilities for optoelectronic devices.
Related JoVE Video
Expression of anterior gradient 2 is decreased with the progression of human biliary tract cancer.
Tohoku J. Exp. Med.
PUBLISHED: 09-05-2014
Show Abstract
Hide Abstract
Biliary tract cancers include cancers of the gallbladder and extrahepatic bile ducts, and its prognosis is poor. The anterior gradient 2 (AGR2) is a protein disulfide isomerase and is highly expressed in various human cancers, such as breast, prostate and pancreas cancers. AGR2 is expressed in normal cholangiocytes and its expression is maintained during biliary carcinogenesis. However, the clinical significance of AGR2 expression in biliary tract cancer has not yet been assessed. Thus, we examined the expression of AGR2 protein in biliary tract tumors using immunohistochemistry and its association with various clinicopathologic parameters. This study included 100 patients who underwent surgery for biliary tract cancers: 46 men and 54 women with a mean and median age of 64.2 and 65.0 years, respectively. AGR2 expression was detected in ductal epithelial cells of the normal biliary tract and in 95% of biliary tract cancer tissues. While the AGR2 expression was not associated with cancer location, patient age, patient sex, degree of regional lymph node metastasis (N-status), or residual status, the AGR2 expression level was decreased with increased tumor size (T-status, p = 0.006) and progression of tumor stage (p = 0.009). Moreover, well-differentiated cancers tended to show higher AGR2 expression than poorly differentiated cancers (p = 0.068); in fact, AGR2 expression was not associated with patient survival (Kaplan-Meier analysis, p = 0.415). Thus, AGR2 is of limited value as a prognostic marker for biliary tract cancer. In conclusion, the expression of AGR2 is decreased with the progression of biliary tract cancer.
Related JoVE Video
Oct4 overexpression facilitates proliferation of porcine fibroblasts and development of cloned embryos.
Zygote
PUBLISHED: 09-02-2014
Show Abstract
Hide Abstract
Summary Octamer-binding transcription factor 4 (Oct4) is a critical molecule for the self-renewal and pluripotency of embryonic stem cells. Recent reports have shown that Oct4 also controls cell-cycle progression and enhances the proliferation of various types of cells. As the high proliferation of donor fibroblasts is critical to the production of transgenic pigs, using the somatic cell nuclear transfer technique, we analysed the effect of Oct4 overexpression on the proliferation of porcine fibroblasts and embryos. Porcine endogenous Oct4 cDNA was cloned, sequenced and inserted into an expression vector. The vector was transfected into porcine fibroblasts, and a stable Oct4-overexpressed cell line was established by antibiotic selection. Oct4 expression was validated by the immunostaining of Oct4. Cell morphology was changed to sharp, and both proliferation and migration abilities were enhanced in Oct4-overexpressed cells. Real-time RT-PCR results showed that p16, Bcl2 and Myc were upregulated in Oct4-overexpressed cells. Somatic cell nuclear transfer was performed using Oct4-overexpressed cells, and the development of Oct4 embryos was compared with that of wild-type cloned embryos. The cleavage and blastocyst formation rates were improved in the Oct4 embryos. Interestingly, blastocyst formation of the Oct4 embryos was observed as early as day 5 in culture, while blastocysts were observed from day 6 in wild-type cloned embryos. In conclusion, the overexpression of Oct4 enhanced the proliferation of both porcine fibroblasts and embryos.
Related JoVE Video
NDRG2 Controls COX-2/PGE2-Mediated Breast Cancer Cell Migration and Invasion.
Mol. Cells
PUBLISHED: 08-19-2014
Show Abstract
Hide Abstract
N-myc downstream-regulated gene 2 (NDRG2), which is known to have tumor suppressor functions, is frequently down-regulated in breast cancers and potentially involved in preventing the migration and invasion of malignant tumor cells. In the present study, we examined the inhibitory effects of NDRG2 overexpression, specifically focusing on the role of cyclooxygenase-2 (COX-2) in the migration of breast cancer cells. NDRG2 overexpression in MDA-MB-231 cells inhibited the expression of the COX-2 mRNA and protein, the transcriptional activity of COX-2, and prostaglandin E2 (PGE2) production, which were induced by a treatment with phorbol-12-myristate-13-acetate (PMA). Nuclear transcription factor-?B (NF-?B) signaling attenuated by NDRG2 expression resulted in a decrease in PMA-induced COX-2 expression. Interestingly, the inhibition of COX-2 strongly suppressed PMA-stimulated migration and invasion in MDA-MB-231-NDRG2 cells. Moreover, siRNA-mediated knockdown of NDRG2 in MCF7 cells increased the COX-2 mRNA and protein expression levels and the PMA-induced COX-2 expression levels. Consistent with these results, the migration and invasion of MCF7 cells treated with NDRG2 siRNA were significantly enhanced following treatment with PMA. Taken together, our data show that the inhibition of NF-?B signaling by NDRG2 expression is able to suppress cell migration and invasion through the down-regulation of COX-2 expression.
Related JoVE Video
Bariatric surgery improves the cavernosal neuronal, vasorelaxation, and contraction mechanisms for erectile dysfunction as result of amelioration of glucose homeostasis in a diabetic rat model.
PLoS ONE
PUBLISHED: 08-19-2014
Show Abstract
Hide Abstract
Bariatric surgery is an effective treatment option for both obesity and obesity-related type 2 diabetes mellitus (T2DM). However, little is known regarding the effects of bariatric surgery on erectile dysfunction among patients with T2DM. Therefore, we investigated whether bariatric surgery would lead to structural and biochemical changes in the corpus cavernosum.
Related JoVE Video
Detection of CTX-II in serum and urine to diagnose osteoarthritis by using a fluoro-microbeads guiding chip.
Biosens Bioelectron
PUBLISHED: 08-17-2014
Show Abstract
Hide Abstract
This study reports a new strategy for simultaneous detection of the C-telopeptide fragments of type II collagen (CTX-II) as a biomarker of osteoarthritis (OA) using a fluoro-microbeads guiding chip. As osteoarthritis progresses, the joint components including matrix and cartilage are degraded by proteases. The degraded products such as CTX-II are released into the serum and urine, and the CTX-II concentration in body fluids reflects OA progression. Because the CTX-II has heterogeneous epitope structure in serum (sCTX-II; homodimers) and urine (uCTX-II; monomers or variant monomers), a multiple-sensing device enabling both sandwich and competitive-type immunoassays is required. For multiple assessments of serum and urinary CTX-II, we designed a fluoro-microbeads guiding chip (FMGC) containing multiple sensing areas and connecting channels. Using the approach, the sandwich (sCTX-II) and competition (uCTX-II) assays could be simultaneously performed on a single chip. We designed a fluidic control device enabling selective control of the open-close function of FMGC channels. The immune-specific signal was quantitatively analyzed by counting the number of fluorescent microbeads from the registered images. The results from the developed FMGC assay showed high correlation with those obtained in ELISA. The completion time of the FMGC assay was 24-fold and 3.5-fold shorter than the ELISA for urinary and serum CTX-II. Taken together, it enabled the simultaneous detection of both sCTX-II and uCTX-II. This FMGC-based assay would be a promising tool for monitoring of osteoarthritis.
Related JoVE Video
Identification and in vivo functional characterization of novel compound heterozygous BMP1 variants in osteogenesis imperfecta.
Hum. Mutat.
PUBLISHED: 07-02-2014
Show Abstract
Hide Abstract
Osteogenesis imperfecta (OI) comprises a heterogeneous group of disorders that are characterized by susceptibility to bone fractures, and range in severity from a subtle increase in fracture frequency to death in the perinatal period. Most patients have defects in type I collagen biosynthesis with autosomal dominant inheritance; but, many autosomal recessive genes have been reported. We applied whole exome sequencing to identify mutations in a Korean OI patient who had an umbilical hernia, frequent fractures, a markedly short stature, delayed motor development, scoliosis, and dislocation of the radial head, with a bowed radius and ulna. We identified two novel variants in the BMP1 gene: c.808A>G and c.1297G>T. The former variant caused a missense change p.(Met270Val) and the latter variant caused the skipping of exon 10. The hypo-functional nature of the two variants was demonstrated in a zebrafish assay. This article is protected by copyright. All rights reserved.
Related JoVE Video
Change in renal function following laparoscopic donor nephrectomy using 99 mTc-diethylenetriaminepentaacetic acid scan.
World J Urol
PUBLISHED: 07-02-2014
Show Abstract
Hide Abstract
We assessed the change in remaining kidney function after laparoscopic donor nephrectomy using serial technetium 99m diethylenetriaminepentaacetic acid (DTPA) scans and investigated the factors affecting the course.
Related JoVE Video
Light-output enhancement of GaN-based vertical light-emitting diodes using periodic and conical nanopillar structures.
Opt Lett
PUBLISHED: 07-01-2014
Show Abstract
Hide Abstract
We investigated GaN-based vertical light-emitting diodes (VLEDs) with periodic and conical nanopillar arrays (CNAs) to improve the light-output efficiency. We found that a 470 nm diameter and 0.8-0.9 ?m height increased the light output, and the devices suffered no significant electrical property degradations. The light-output power was 272% and 5.1% greater than flat- and rough-surface VLEDs at 350 mA, respectively. These improved optical properties are attributed to the optimized CNAs, which increase the effective photon escape cone and reduce the total internal reflection at the n-GaN-air interface. We also investigated the emission characteristics and mechanisms with finite-difference time-domain simulations.
Related JoVE Video
Genome-wide identification of nuclear receptor (NR) superfamily genes in the copepod Tigriopus japonicus.
BMC Genomics
PUBLISHED: 06-20-2014
Show Abstract
Hide Abstract
Nuclear receptors (NRs) are a large superfamily of proteins defined by a DNA-binding domain (DBD) and a ligand-binding domain (LBD). They function as transcriptional regulators to control expression of genes involved in development, homeostasis, and metabolism. The number of NRs differs from species to species, because of gene duplications and/or lineage-specific gene losses during metazoan evolution. Many NRs in arthropods interact with the ecdysteroid hormone and are involved in ecdysone-mediated signaling in arthropods. The nuclear receptor superfamily complement has been reported in several arthropods, including crustaceans, but not in copepods. We identified the entire NR repertoire of the copepod Tigriopus japonicus, which is an important marine model species for ecotoxicology and environmental genomics.
Related JoVE Video
Knockdown of anterior gradient 2 expression extenuates tumor-associated phenotypes of SNU-478 ampulla of Vater cancer cells.
BMC Cancer
PUBLISHED: 06-18-2014
Show Abstract
Hide Abstract
Anterior gradient 2 (AGR2) has been implicated in tumor-associated phenotypes such as cell viability, invasion and metastasis in various human cancers. However, the tumor promoting activity of AGR2 has not yet been determined in biliary tract cancers. Thus, we examined the expression of AGR2 and its tumor-promoting activity in biliary tract cancer cells in this study.
Related JoVE Video
Engineering a horseradish peroxidase C stable to radical attacks by mutating multiple radical coupling sites.
Biotechnol. Bioeng.
PUBLISHED: 06-09-2014
Show Abstract
Hide Abstract
Peroxidases have great potential as industrial biocatalysts. In particular, the oxidative polymerization of phenolic compounds catalyzed by peroxidases has been extensively examined because of the advantage of this method over other conventional chemical methods. However, the industrial application of peroxidases is often limited because of their rapid inactivation by phenoxyl radicals during oxidative polymerization. In this work, we report a novel protein engineering approach to improve the radical stability of horseradish peroxidase isozyme C (HRPC). Phenylalanine residues that are vulnerable to modification by the phenoxyl radicals were identified using mass spectrometry analysis. UV-Vis and CD spectra showed that radical coupling did not change the secondary structure or the active site of HRPC. Four phenylalanine (Phe) residues (F68, F142, F143, and F179) were each mutated to alanine residues to generate single mutants to examine the role of these sites in radical coupling. Despite marginal improvement of radical stability, each single mutant still exhibited rapid radical inactivation. To further reduce inactivation by radical coupling, the four substitution mutations were combined in F68A/F142A/F143A/F179A. This mutant demonstrated dramatic enhancement of radical stability by retaining 41% of its initial activity compared to the wild-type, which was completely inactivated. Structure and sequence alignment revealed that radical vulnerable Phe residues of HPRC are conserved in homologous peroxidases, which showed the same rapid inactivation tendency as HRPC. Based on our site-directed mutagenesis and biochemical characterization, we have shown that engineering radical-vulnerable residues to eliminate multiple radical coupling can be a good strategy to improve the stability of peroxidases against radical attack. Biotechnol. Bioeng. © 2014 Wiley Periodicals, Inc.
Related JoVE Video
The inhibitory effect of an ethanol extract of the spores of Lygodium japonicum on ethylene glycol-induced kidney calculi in rats.
Urolithiasis
PUBLISHED: 06-09-2014
Show Abstract
Hide Abstract
We investigated the effect of an ethanol extract of Lygodii spora (LS) as a preventive and therapeutic agent for experimentally induced calcium oxalate nephrolithiasis with ethylene glycol (EG) in rats. Male Wistar rats were randomly divided into preventive (n = 18, for 28 days) and therapeutic (n = 24, for 42 days) groups. The preventive group was further subdivided into three groups of six rats each: preventive control, preventive lithiatic control (EG) and preventive lithiatic LS (EG + 400 mg/kg LS). Similarly, the therapeutic group was subdivided into four groups of six rats each: therapeutic control, therapeutic lithiatic control, therapeutic lithiatic untreated, and therapeutic lithiatic LS. Lithiasis was induced by adding 0.75% EG to the drinking water of all groups except the preventive and therapeutic control groups. Preventive and therapeutic subjects also received the LS ethanol extract in drinking water at a dose of 400 mg/kg, since day 0 or day 28, respectively. At the end of the each experimental period, various biochemical parameters were measured in urine and kidney homogenates. The kidneys were subjected to histopathological analysis. The results revealed that treatment with the LS preventive protocol significantly decreased the levels of urinary calcium, oxalate and uric acid, and increased the levels of urinary citrate as compared to those in the EG control. No significant changes in the urinary parameters except oxalate and citrate levels were observed in the rats in the therapeutic protocol. In both preventive and therapeutic protocols, the extract significantly decreased kidney peroxides, renal calcium, oxalate content, and the number of kidney oxalate deposits as compared to those in the EG group. We conclude that LS is useful as a preventive and therapeutic agent against the formation of oxalate kidney stones.
Related JoVE Video
Recent advances in parametric neuroreceptor mapping with dynamic PET: basic concepts and graphical analyses.
Neurosci Bull
PUBLISHED: 06-08-2014
Show Abstract
Hide Abstract
Tracer kinetic modeling in dynamic positron emission tomography (PET) has been widely used to investigate the characteristic distribution patterns or dysfunctions of neuroreceptors in brain diseases. Its practical goal has progressed from regional data quantification to parametric mapping that produces images of kinetic-model parameters by fully exploiting the spatiotemporal information in dynamic PET data. Graphical analysis (GA) is a major parametric mapping technique that is independent on any compartmental model configuration, robust to noise, and computationally efficient. In this paper, we provide an overview of recent advances in the parametric mapping of neuroreceptor binding based on GA methods. The associated basic concepts in tracer kinetic modeling are presented, including commonly-used compartment models and major parameters of interest. Technical details of GA approaches for reversible and irreversible radioligands are described, considering both plasma input and reference tissue input models. Their statistical properties are discussed in view of parametric imaging.
Related JoVE Video
Effects of KH-204 on the expression of heat shock protein 70 and germ cell apoptosis in infertility rat models.
BMC Complement Altern Med
PUBLISHED: 06-05-2014
Show Abstract
Hide Abstract
Idiopathic infertility is a significant number of causes of male infertility. Empirical treatments are used for idiopathic male infertility, and antioxidant supplementation is a kind of management of oxidative stress related infertility. We investigated the antioxidant effects of the modified Ojayeonjonghwan (KH-204) in a rat model of cryptorchidism.
Related JoVE Video
Supramolecular Hydrogels for Long-Term Bioengineered Stem Cell Therapy.
Adv Healthc Mater
PUBLISHED: 06-03-2014
Show Abstract
Hide Abstract
Synthetic hydrogels have been extensively investigated as artificial extracellular matrices (ECMs) for tissue engineering in vitro and in vivo. Crucial challenges for such hydrogels are sustaining long-term cytocompatible encapsulation and providing appropriate cues at the right place and time for spatio-temporal control of the cells. Here, in situ supramolecularly assembled and modularly modified hydrogels for long-term engineered mesenchymal stem cell (eMSC) therapy are reported using cucurbit[6]uril-conjugated hyaluronic acid (CB[6]-HA), diaminohexane conjugated HA (DAH-HA), and drug-conjugated CB[6] (drug-CB[6]). The eMSCs producing enhanced green fluorescence protein (EGFP) remain alive and emit the fluorescence within CB[6]/DAH-HA hydrogels in mice for more than 60 d. Furthermore, the long-term expression of mutant interleukin-12 (IL-12M) by eMSCs within the supramolecular hydrogels results in effective inhibition of tumor growth with a significantly enhanced survival rate. Taken together, these findings confirm the feasibility of supramolecular HA hydrogels as 3D artificial ECMs for cell therapies and tissue engineering applications.
Related JoVE Video
Development of the radical-stable Coprinus cinereus peroxidase (CiP) by blocking the radical attack.
J. Biotechnol.
PUBLISHED: 05-22-2014
Show Abstract
Hide Abstract
Despite the potential use of peroxidases as industrial biocatalysts, their practical application is often impeded due to suicide inactivation by radicals generated in oxidative reactions. Using a peroxidase from Coprinus cinereus (CiP) as a model enzyme, we revealed a dominant factor for peroxidase inactivation during phenol oxidation, and we engineered radical-stable mutants by site-directed mutagenesis of an amino acid residue susceptible to modification by phenoxyl radical. Mass spectrometry analysis of inactivated CiP identified an adduct between F230 and a phenoxyl radical, and subsequently, the F230 residue was mutated to amino acids that resisted radical coupling. Of the F230 mutants, the F230A mutant showed the highest stability against radical inactivation, retaining 80% of its initial activity, while the wild-type protein was almost completely inactivated. The F230A mutant also exhibited a 16-fold higher turnover of the phenol substrate compared with the wild-type enzyme. Furthermore, the F230A mutant was stable during the oxidation of other phenolic compounds, including m-cresol and 3-methoxyphenol. No structural changes were observed by UV-vis and CD spectra of CiP after radical coupling, implying that the F230-phenol radical adduct inactivated CiP by blocking substrate access to the active site. Our novel strategy can be used to improve the stability of other peroxidases inactivated by radicals.
Related JoVE Video
An Improved Antagonist Radiotracer for the ?-Opioid Receptor: Synthesis and Characterization of 11C-LY2459989.
J. Nucl. Med.
PUBLISHED: 05-22-2014
Show Abstract
Hide Abstract
The ?-opioid receptors (KORs) are implicated in several neuropsychiatric diseases and addictive disorders. PET with radioligands provides a means to image the KOR in vivo and investigate its function in health and disease. The purpose of this study was to develop the selective KOR antagonist (11)C-LY2459989 as a PET radioligand and characterize its imaging performance in nonhuman primates.
Related JoVE Video
Cellular Toxicity of Calf Blood Extract on Human Corneal Epithelial Cells In Vitro.
Curr. Eye Res.
PUBLISHED: 05-17-2014
Show Abstract
Hide Abstract
Abstract Purpose: To investigate the biologic effects of the calf blood extract on corneal epithelial cells in vitro. Materials and methods: The effects on corneal epithelial cells were evaluated after 1, 4, 12, and 24?h of exposure to various concentrations of calf blood extract (3, 5, 8 and 16%). The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay was performed to measure levels of cellular metabolic activity. The lactate dehydrogenase (LDH) assay was performed to determine the extent of cellular damage. Cellular morphology was examined using phase-contrast microscopy. The scratch wound assay was performed to quantify the migration of corneal epithelial cells. Results: At the 3 and 5% concentrations of calf blood extract, MTT values were similar to those observed in the control group. However, at a concentration of 8 and 16%, cellular metabolic activity was significantly decreased after 4?h of exposure to calf blood extract. After 12?h of exposure to 8 and 16% concentrations of calf blood extract, LDH activity and cellular morphological damage to the corneal epithelial cells were significantly increased. There was no evidence of cellular migration after 12?h exposure to 5% or higher concentration of calf blood extract because of cellular toxicity. Conclusions: Compared with normal corneal epithelial cells, the cellular activity was decreased, and toxicity was increased after over 12?h of exposure to more than 5% concentration of calf blood extract. Further clinical studies will be necessary to determine the optimal concentration and exposure time for the topical application of eye drops containing calf blood extract.
Related JoVE Video
Kazinol C from Broussonetia kazinoki activates AMP-activated protein kinase to induce antitumorigenic effects in HT-29 colon cancer cells.
Oncol. Rep.
PUBLISHED: 05-15-2014
Show Abstract
Hide Abstract
Kazinol C is a 1,3-diphenylpropane, obtained from Broussonetia kazinoki, that has been employed in folk medicine as an edema suppressant. It exerts beneficial effects in oxidative stress-related diseases, such as cancer. However, the molecular mechanism involved in the anticancer effects remains to be determined. AMP-activated protein kinase (AMPK) has emerged as a possible anticancer target molecule. The present study investigated the effect of kazinol C on AMPK activation as well as subsequent HT-29 colon cancer cell viability, apoptosis and migration. Kazinol C markedly induced AMPK phosphorylation and significantly attenuated HT-29 colon cancer cell growth and viability. Compound C, as a well?known AMPK inhibitor, blocked the kazinol C-induced cell death, and stable transduction of dominant-negative (DN) AMPK in colon cancer cells also inhibited kazinol C-induced cell apoptosis. In addition, kazinol C inhibited HT-29 cell migration and anchorage-independent growth. AMPK inhibition using stable transduction with DN AMPK significantly abrogated the kazinol C-induced inhibition of cancer cell migration. Thus, AMPK is a critical and novel regulator of kazinol C-mediated cancer cell apoptosis and inhibition of migration, suggesting that AMPK is a prime cancer target.
Related JoVE Video
Fabrication of wide-bandgap transparent electrodes by using conductive filaments: performance breakthrough in vertical-type GaN LED.
Sci Rep
PUBLISHED: 05-07-2014
Show Abstract
Hide Abstract
For realizing next-generation solid-state lighting devices, performance breakthroughs must be accomplished for nitride-based light-emitting diodes (LEDs). Highly transparent conductive electrodes (TCEs) may be key to achieving this goal, as they provide uniform current injection and distribution across a large device area, eventually increasing the light output power. However, the trade-off between electrical conductivity and optical transmittance of LEDs must be addressed. Herein, we introduce a novel strategy based on TCEs fabricated using wide-bandgap (WB) materials such as SiNx, incorporated beneath the n-type electrode of vertical-type LEDs, and show the feasibility of this strategy. We employ a novel electrical breakdown (EBD) technique to form conductive filaments (or current paths) between a TCE and n-GaN (GaN: gallium nitride). By employing the EBD process, we obtain both ohmic behavior for SiNx TCE/n-GaN and a current spreading effect across n-GaN. These results demonstrate the tremendous potential of WB-TCEs for use in high-performance optoelectronic devices.
Related JoVE Video
Improvement of cloned embryos development by co-culturing with parthenotes: a possible role of exosomes/microvesicles for embryos paracrine communication.
Cell Reprogram
PUBLISHED: 04-28-2014
Show Abstract
Hide Abstract
It is well known that embryos cultured in a group can create a microenvironment through secretion of autocrine and paracrine factors that can support and improve the embryos' development when compared to the embryos cultured individually. In this study, we used a co-culture system for paracrine communication between different kinds of embryos. The results showed that co-culture of porcine parthenogenetic (PA) embryos significantly improved the in vitro development of cloned (nuclear transfer, NT) embryos. To reveal the possible mechanism of communication between the two groups, we isolated exosomes/microvesicles (EXs/MVs) from the PA embryos conditioned medium (PA-CM) through differential centrifugation and identified them through transmission electron microscope and immunoflourescence against exosomal/membrane marker CD9. Furthermore, these EXs/MVs were found to contain mRNA of pluripotency genes (Oct4, Sox2, Klf4, c-Myc, and Nanog), and the PKH67-labeled EXs/MVs could be internalized by the NT embryos. The current study demonstrates that cloned embryos' developmental competence can be improved through co-culturing with PA embryos and revealed, for the first time, that in vitro-produced embryos can secrete EXs/MVs as a possible communication tool within their microenvironment. Moreover, it provides a new paradigm for embryo-to-embryo communication in vitro.
Related JoVE Video
NDRG2 overexpression enhances glucose deprivation-mediated apoptosis in breast cancer cells via inhibition of the LKB1-AMPK pathway.
Genes Cancer
PUBLISHED: 04-04-2014
Show Abstract
Hide Abstract
The newly identified tumor suppressor, N-myc downstream-regulated gene 2 (NDRG2), has been studied in various cancers because of its anticancer and antimetastasis effects. In this study, we examined the effect of NDRG2 expression on cell viability in MDA-MB-231 human breast cancer cells under conditions that are similar to the microenvironment of solid tumors, which include glucose deprivation. NDRG2 overexpression enhanced the pro-apoptotic effects of glucose deprivation. Glucose deprivation also induced the activation of AMP-activated protein kinase (AMPK), which plays a role in protecting tumor cells from metabolic stresses. NDRG2 overexpression strongly reduced glucose deprivation-induced AMPK phosphorylation and increased the cleavage of poly (ADP-ribose) polymerase (PARP), which indicated the induction of apoptosis. The expression of a constitutively active form of AMPK effectively blocked glucose deprivation-induced apoptosis in NDRG2-overexpressing MDA-MB-231 cells. Moreover, NDRG2 overexpression also enhanced the pro-apoptotic effects of 2-deoxyglucose (2-DG) or hypoxia, an inducer of metabolic stresses. Finally, we showed that LKB1 is an upstream kinase of AMPK that is involved in the inhibition of glucose deprivation-induced AMPK activity in NDRG2-overexpressing cells. Our findings collectively suggest that NDRG2 is a negative regulator of AMPK activity and functions as a sensitizer of glucose deprivation.
Related JoVE Video
Crystallization and preliminary X-ray crystallographic analysis of the complex between the N-D1 domain of VCP from Homo sapiens and the N domain of OTU1 from Saccharomyces cerevisiae.
Acta Crystallogr F Struct Biol Commun
PUBLISHED: 03-27-2014
Show Abstract
Hide Abstract
VCP (valosin-containing protein; also known as p97) plays important roles in many biological processes including the ERAD (endoplasmic reticulum-associated degradation) pathway and its function is governed by binding partners. OTU1 (ovarian tumour domain-containing protein 1) is a recently discovered deubiquitinating enzyme that interacts directly with VCP in the ERAD pathway. In order to understand the interactions between the two proteins, the N-D1 domain of VCP and the UBXL domain of OTU1 were cloned, overexpressed, purified and crystallized. The crystals of the complex diffracted to 3.25?Å resolution and belonged to space group P21, with unit-cell parameters a = 165.45, b = 176.73, c = 165.59?Å, ? = 120.095°. There are two molecules of the complex in the asymmetric unit with a Matthews coefficient of 2.62?Å(3)?Da(-1) and a solvent content of 53%.
Related JoVE Video
Production of CMAH Knockout Preimplantation Embryos Derived From Immortalized Porcine Cells Via TALE Nucleases.
Mol Ther Nucleic Acids
PUBLISHED: 03-18-2014
Show Abstract
Hide Abstract
Although noncancerous immortalized cell lines have been developed by introducing genes into human and murine somatic cells, such cell lines have not been available in large domesticated animals like pigs. For immortalizing porcine cells, primary porcine fetal fibroblasts were isolated and cultured using the human telomerase reverse transcriptase (hTERT) gene. After selecting cells with neomycin for 2 weeks, outgrowing colonized cells were picked up and subcultured for expansion. Immortalized cells were cultured for more than 9 months without changing their doubling time (~24 hours) or their diameter (< 20 µm) while control cells became replicatively senescent during the same period. Even a single cell expanded to confluence in 100?mm dishes. Furthermore, to knockout the CMAH gene, designed plasmids encoding a transcription activator-like effector nuclease (TALENs) pairs were transfected into the immortalized cells. Each single colony was analyzed by the mutation-sensitive T7 endonuclease I assay, fluorescent PCR, and dideoxy sequencing to obtain three independent clonal populations of cells that contained biallelic modifications. One CMAH knockout clone was chosen and used for somatic cell nuclear transfer. Cloned embryos developed to the blastocyst stage. In conclusion, we demonstrated that immortalized porcine fibroblasts were successfully established using the human hTERT gene, and the TALENs enabled biallelic gene disruptions in these immortalized cells.
Related JoVE Video
Peptoniphilus rhinitidis sp. nov., isolated from specimens of chronic rhinosinusitis.
Anaerobe
PUBLISHED: 03-10-2014
Show Abstract
Hide Abstract
Chronic rhinosinusitis (CRS) is an inflammatory disorder of the nasal cavity and paranasal sinus related to bacterial infection. A previous study suggested that a specific bacterial group may have an important role in the course of CRS. In this study, bacteria isolated from CRS patients were characterized. A total of 15 strains were identified as Gram-positive anaerobic cocci (GPAC), which were able to utilize peptone as a sole carbon source. Sequencing of the 16S ribosomal RNA gene revealed that the isolates were closely related to members of the genus Peptoniphilus (>97% similarity) within the Clostridiales Family XI. Incertae Sedis. Genotypic and phenotypic characterization suggests that these isolates represent a novel species of the genus Peptoniphilus associated with CRS. The type strain of Peptoniphilus rhinitidis is 1-13(T) (=KCTC 5985(T) = JCM 17448(T)).
Related JoVE Video
Structural basis for ovarian tumor domain-containing protein 1 (OTU1) binding to p97/valosin-containing protein (VCP).
J. Biol. Chem.
PUBLISHED: 03-07-2014
Show Abstract
Hide Abstract
Valosin-containing protein (VCP), also known as p97, is an AAA(+) ATPase that plays an essential role in a broad array of cellular processes including the endoplasmic reticulum-associated degradation (ERAD) pathway. Recently, ERAD-specific deubiquitinating enzymes have been reported to be physically associated with VCP, although the exact mechanism is not yet clear. Among these enzymes is ovarian tumor domain-containing protein 1 (OTU1). Here, we report the structural basis for interaction between VCP and OTU1. The crystal structure of the ubiquitin regulatory X-like (UBXL) domain of OTU1 (UBXLOTU1) complexed to the N-terminal domain of VCP (NVCP) at 1.8-? resolution reveals that UBXLOTU1 adopts a ubiquitin-like fold and binds at the interface of two subdomains of NVCP using the (39)GYPP(42) loop of UBXLOTU1 with the two prolines in cis- and trans-configurations, respectively. A mutagenesis study shows that this loop is not only critical for the interaction with VCP but also for its role in the ERAD pathway. Negative staining EM shows that one molecule of OTU1 binds to one VCP hexamer, and isothermal titration calorimetry suggests that the two proteins bind with a KD of 0.71 ?M. Analytical size exclusion chromatography and isothermal titration calorimetry demonstrates that OTU1 can bind VCP in both the presence and absence of a heterodimer formed by ubiquitin fusion degradation protein 1 and nuclear localization protein 4.
Related JoVE Video
AMPK?2 translocates into the nucleus and interacts with hnRNP H: implications in metformin-mediated glucose uptake.
Cell. Signal.
PUBLISHED: 02-27-2014
Show Abstract
Hide Abstract
Adenosine monophosphate (AMP)-activated protein kinase (AMPK) is a cytoplasmic protein that plays a critical role in the maintenance of energy homeostasis. However, its role in the nucleus is still largely unknown. Here, we showed that AMPK?2 translocated into the nucleus during muscle differentiation. We also showed that upon treatment with 5-aminoimidazole-4-carboxy-amide-1-d-ribofuranoside (AICAR), an AMPK activator, AMPK rapidly translocated into the nucleus in rat myoblast L6 cells. On the other hand, the AMPK?2 phosphorylation-defective mutant did not translocate into the nucleus. Knockdown of AMPK?2 suppressed the differentiation-induced expression of myogenin, a differentiation marker. A physiological AMPK activator, metformin, also induced the translocation of AMPK?2 into the nucleus. Both inhibition and knockdown of AMPK?2 suppressed metformin-mediated glucose uptake. In addition, AMPK?2 was shown to directly interact with the heterogeneous nuclear ribonucleoprotein H (hnRNP H). AICAR treatment increased the phosphorylation of hnRNP H. Metformin increased the interaction between AMPK?2 and hnRNP H in the nucleus. Knockdown of hnRNP H blocked metformin-induced glucose uptake. In summary, these results demonstrate that AMPK?2 translocates into the nucleus via phosphorylation, AMPK?2 interacts with and phosphorylates hnRNP H in the nucleus, and such a protein-protein interaction modulates metformin-mediated glucose uptake.
Related JoVE Video
Production and characterization of soluble human TNFRI-Fc and human HO-1(HMOX1) transgenic pigs by using the F2A peptide.
Transgenic Res.
PUBLISHED: 02-05-2014
Show Abstract
Hide Abstract
Generation of transgenic pigs for xenotransplantation is one of the most promising technologies for resolving organ shortages. Human heme oxygenase-1 (hHO-1/HMOX1) can protect transplanted organs by its strong anti-oxidative, anti-apoptotic, and anti-inflammatory effects. Soluble human TNFRI-Fc (shTNFRI-Fc) can inhibit the binding of human TNF-? (hTNF-?) to TNF receptors on porcine cells, and thereby, prevent hTNF-?-mediated inflammation and apoptosis. Herein, we successfully generated shTNFRI-Fc-F2A-HA-hHO-1 transgenic (TG) pigs expressing both shTNFRI-Fc and hemagglutinin-tagged-human heme oxygenase-1 (HA-hHO-1) by using an F2A self-cleaving peptide. shTNFRI-Fc and HA-hHO-1 transgenes containing the F2A peptide were constructed under the control of the CAG promoter. Transgene insertion and copy number in the genome of transgenic pigs was confirmed by polymerase chain reaction (PCR) and Southern blot analysis. Expressions of shTNFRI-Fc and HA-hHO-1 in TG pigs were confirmed using PCR, RT-PCR, western blot, ELISA, and immunohistochemistry. shTNFRI-Fc and HA-hHO-1 were expressed in various organs, including the heart, lung, and spleen. ELISA assays detected shTNFRI-Fc in the sera of TG pigs. For functional analysis, fibroblasts isolated from a shTNFRI-Fc-F2A-HA-hHO-1 TG pig (i.e., #14; 1 × 10(5) cells) were cultured with hTNF-? (20 ng/mL) and cycloheximide (10 ?g/mL). The viability of shTNFRI-Fc-F2A-HA-hHO-1 TG pig fibroblasts was significantly higher than that of the wild type (wild type vs. shTNFRI-Fc-F2A-HA-hHO-1 TG at 24 h, 31.6 ± 3.2 vs. 60.4 ± 8.3 %, respectively; p < 0.05). Caspase-3/-7 activity of the shTNFRI-Fc-F2A-HA-hHO-1 TG pig fibroblasts was lower than that of the wild type pig fibroblasts (wild type vs. shTNFRI-Fc-F2A-HA-hHO-1 TG at 12 h, 812,452 ± 113,078 RLU vs. 88,240 ± 10,438 RLU, respectively; p < 0.05). These results show that shTNFRI-Fc and HA-hHO-1 TG pigs generated by the F2A self-cleaving peptide express both shTNFRI-Fc and HA-hHO-1 molecules, which provides protection against oxidative and inflammatory injury. Utilization of the F2A self-cleaving peptide is a promising tool for generating multiple TG pigs for xenotransplantation.
Related JoVE Video
Impact of enzyme replacement therapy on linear growth in Korean patients with mucopolysaccharidosis type II (Hunter syndrome).
J. Korean Med. Sci.
PUBLISHED: 01-28-2014
Show Abstract
Hide Abstract
Hunter syndrome (or mucopolysaccharidosis type II [MPS II]) arises because of a deficiency in the lysosomal enzyme iduronate-2-sulfatase. Short stature is a prominent and consistent feature in MPS II. Enzyme replacement therapy (ERT) with idursulfase (Elaprase®) or idursulfase beta (Hunterase®) have been developed for these patients. The effect of ERT on the growth of Korean patients with Hunter syndrome was evaluated at a single center. This study comprised 32 patients, who had received ERT for at least 2 yr; they were divided into three groups according to their ages at the start of ERT: group 1 (<6 yr, n=14), group 2 (6-10 yr, n=11), and group 3 (10-20 yr, n=7). The patients showed marked growth retardation as they got older. ERT may have less effect on the growth of patients with the severe form of Hunter syndrome. The height z-scores in groups 2 and 3 revealed a significant change (the estimated slopes before and after the treatment were -0.047 and -0.007, respectively: difference in the slope, 0.04; P<0.001). Growth in response to ERT could be an important treatment outcome or an endpoint for future studies.
Related JoVE Video
Vav3, a GEF for RhoA, Plays a Critical Role under High Glucose Conditions.
Endocrinol Metab (Seoul)
PUBLISHED: 01-27-2014
Show Abstract
Hide Abstract
The role of small GTPase molecules is poorly understood under high glucose conditions.
Related JoVE Video
Clinical outcomes of second-look endoscopy after gastric endoscopic submucosal dissection: predictive factors with high risks of bleeding.
Surg Endosc
PUBLISHED: 01-22-2014
Show Abstract
Hide Abstract
A second-look endoscopy is routinely performed after endoscopic submucosal dissection (ESD) in many institutions, although the need is questionable. Additional hemostatic procedures might be necessary for the post-ESD ulcer with a high risk of bleeding. We investigated the predictive factors for post-ESD ulcers with a high risk of bleeding.
Related JoVE Video
Voxelwise lp-ntPET for detecting localized, transient dopamine release of unknown timing: sensitivity analysis and application to cigarette smoking in the PET scanner.
Hum Brain Mapp
PUBLISHED: 01-20-2014
Show Abstract
Hide Abstract
The "linear parametric neurotransmitter PET" (lp-ntPET) model estimates time variation in endogenous neurotransmitter levels from dynamic PET data. The pattern of dopamine (DA) change over time may be an important element of the brain's response to addictive substances such as cigarettes or alcohol. We have extended the lp-ntPET model from the original region of interest (ROI) - based implementation to be able to apply the model at the voxel level. The resulting endpoint is a dynamic image, or movie, of transient neurotransmitter changes. Simulations were performed to select threshold values to reduce the false positive rate when applied to real (11)C-raclopride PET data. We tested the new voxelwise method on simulated data, and finally, we applied it to (11)C-raclopride PET data of subjects smoking cigarettes in the PET scanner. In simulation, the temporal precision of neurotransmitter response was shown to be similar to that of ROI-based lp-ntPET (standard deviation ? 3 min). False positive rates for the voxelwise method were well controlled by combining a statistical threshold (the F-test) with a new spatial (cluster-size) thresholding operation. Sensitivity of detection for the new algorithm was greater than 80% for the case of short-lived DA changes that occur in subregions of the striatum as might be the case with cigarette smoking. Finally, in (11)C-raclopride PET data, DA movies reveal for the first time that different temporal patterns of the DA response to smoking may exist in different subregions of the striatum. These spatiotemporal patterns of neurotransmitter change created by voxelwise lp-ntPET may serve as novel biomarkers for addiction and/or treatment efficacy.
Related JoVE Video
First thermostable endo-?-1,4-glucanase from newly isolated Xanthomonas sp. EC102.
Protein J.
PUBLISHED: 01-09-2014
Show Abstract
Hide Abstract
A novel gene encoding thermostable endoglucanase was identified in Xanthomonas sp. EC102 from soil. The gene had 1,458 base pairs of open reading frame, which encode a 52-kDa protein of 486 amino acid residues. Sequence of the amino acid residues was similar with the endoglucanase from Xanthomonas campestris pv. campestris ATCC33913 (GenBank Accession No. NP_638867.1) (94 % identity). The endoglucanase was overexpressed in Escherichia coli BL21 and purified. Temperature for the highest enzymatic activity was 70 °C and pH optima was pH 5.5. The specific activity of the endoglucanase toward carboxymethylcellulose (CMC) was approximately 2 ?mol min?¹ mg?¹, V max for CMC was 1.44 ?mol mg?¹ min?¹, and K m values was 25.6 mg mL?¹. The EC102 endoglucanase was stable at temperatures up to 60 °C, and it was activated by 0.1 mM of Mn²? and Co²?. This is the first report about thermostable endoglucanase from Xanthomonas sp.
Related JoVE Video
Identification of novel hypermethylated genes and demethylating effect of vincristine in colorectal cancer.
J. Exp. Clin. Cancer Res.
PUBLISHED: 01-02-2014
Show Abstract
Hide Abstract
Colorectal cancer (CRC) arises as a consequence of genetic events such as gene mutation and epigenetic alteration. The aim of this study was to identify new hypermethylated candidate genes and methylation-based therapeutic targets using vincristine in CRC.
Related JoVE Video
Silver Wire Amplifies the Signaling Mechanism for IL-1beta Production More Than Silver Submicroparticles in Human Monocytic THP-1 Cells.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Silver materials have been widely used in diverse fields. However, their toxicity and their mechanism, especially in different forms, have not been studied sufficiently. Thus, cytotoxicity, apoptosis, and interleukin-1beta (IL-1?) production were investigated using macrophage-like THP-1 cells in the presence of Ag microparticles (AgMPs, 2.7 µm), Ag submicroparticles (AgSMPs, 150 nm), and Ag wires (AgWs, 274 nm×5.3 µm). The levels of cytotoxicity, apoptosis, and IL-1? production by AgWs were higher than those by the other two AgSMPs and AgMPs. This trend was also observed with each step of the signaling mechanism for IL-1? production, which is a single pathway affiliated with ROS generation or lysosomal rupture or both, cathepsin B, caspase-1 (NALP3 inflammasome), and finally IL-1? production in THP-1 cells. All these results suggest that, for development of safe and effective silver materials, the shape or form of silver materials should be considered, especially for macrophage cell lines because epithelial cell lines are not overly sensitive to silver materials.
Related JoVE Video
The effects of new herbal formula (KBMSI-2) on penile erection and expression of nitric oxide synthase isoforms in streptozotocin-induced diabetic rat model.
Chin J Integr Med
PUBLISHED: 11-16-2013
Show Abstract
Hide Abstract
To investigate the effects of new herbal formula (KBMSI-2) on erectile dysfunction in streptozotocin (STZ)-induced diabetic rat model.
Related JoVE Video
A randomised controlled trial comparing a thermal massager with artificial teardrops for the treatment of dry eye.
Br J Ophthalmol
PUBLISHED: 10-16-2013
Show Abstract
Hide Abstract
To evaluate the efficacy and safety of a thermal massager for the treatment of dry eye syndrome.
Related JoVE Video
Clinical bacteriology of recurrent otitis media with effusion.
Acta Otolaryngol.
PUBLISHED: 10-16-2013
Show Abstract
Hide Abstract
Staphylococcus species were the most common bacterial isolates from patients with otitis media with effusion (OME). Unexpectedly, however, there was no difference in the identity or antibiotic resistance of bacteria isolated from patients with recurrent and non-recurrent OME.
Related JoVE Video
The anti-inflammatory and antimicrobial effects of anthocyanin extracted from black soybean on chronic bacterial prostatitis rat model.
Chin J Integr Med
PUBLISHED: 10-14-2013
Show Abstract
Hide Abstract
To investigated the anti-inflammatory and antimicrobial effects of anthocyanins extracted from black soybean on the chronic bacterial prostatitis (CBP) rat model.
Related JoVE Video
Duodenal mucosa-associated lymphoid tissue lymphomas: two cases and the evaluation of endoscopic ultrasonography.
Clin Endosc
PUBLISHED: 09-30-2013
Show Abstract
Hide Abstract
Mucosa-associated lymphoid tissue lymphoma mainly arises in the stomach, with fewer than 30% arising in the small intestine. We describe here two cases of primary duodenal mucosa-associated lymphoid tissue lymphoma which were evaluated by endoscopic ultrasonography. A 52-year-old man underwent endoscopy due to abdominal pain, which demonstrated a depressed lesion on duodenal bulb. Endoscopic ultrasonographic finding was hypoechoic lesion invading the submucosa. The other case was a previously healthy 51-year-old man. Endoscopy showed a whitish granular lesion on duodenum third portion. Endoscopic ultrasonography image was similar to the first case, whereas abdominal computed tomography revealed enlargement of multiple lymph nodes. The first case was treated with eradication of Helicobacter pylori, after which the mucosal change and endoscopic ultrasound finding were normalized in 7 months. The second case was treated with cyclophosphamide, vincristine, prednisolone, and rituximab every 3 weeks. After 6 courses of chemotherapy, the patient achieved complete remission.
Related JoVE Video
Fruit quality, anthocyanin and total phenolic contents, and antioxidant activities of 45 blueberry cultivars grown in Suwon, Korea.
J Zhejiang Univ Sci B
PUBLISHED: 09-07-2013
Show Abstract
Hide Abstract
Blueberry fruits from 45 commercial cultivars (39 northern highbush and 6 half highbush blueberry) grown in Suwon, Korea were analyzed for fruit size, soluble solids content, titratable acidity, total anthocyanin content, total phenolic content, and antioxidant activity. Fruit characteristics varied widely among the 45 blueberry cultivars. Fruit weight ranged from 0.9 to 3.6 g, soluble solids content from 8.3 to 14.3 °Brix, and titratable acidity from 0.8% to 3.6%. Antioxidant activity ranged from 0.7 to 2.1 mg of quercetin equivalents per gram of fresh berries in different blueberry cultivars. Among the 45 blueberry cultivars, high amounts of anthocyanins and polyphenols, and high antioxidant activity were observed in Elliott, Rubel, Rancocas, and Friendship.
Related JoVE Video
Mitogen-activated protein kinase phosphatase-1 inhibition and sustained extracellular signal-regulated kinase 1/2 activation in camptothecin-induced human colon cancer cell death.
Cancer Biol. Ther.
PUBLISHED: 08-28-2013
Show Abstract
Hide Abstract
Camptothecins are commonly used chemotherapeutics; in some models, they enhance signaling via the mitogen-activated protein kinase (MAPK) pathway through effects on upstream kinases. To evaluate the impact of camptothecin (CPT) on MAPKs in human colon cancer, we studied HCT116 and CaCo2 colon cancer cells. We found that HCT116 cells highly express mitogen-activated protein kinase phosphatase-1 (MKP1), which selectively inactivates extracellular signal-regulated kinase (ERK), whereas MKP1 levels were undetectable in CaCo2 cells. CPT did not affect ERK activity in CaCo2 cells, but did induce a striking increase in ERK activity in HCT116 cells in association with a corresponding decrease in MKP1. The reduction in MKP1 expression occurred at a posttranscriptional level and was blocked by the proteasome inhibitor MG132, whereas that CPT-induced downregulation of MKP1 was not due to proteasome-mediated degradation. Treatment of HCT116 cells with CPT induced a sustained activation of nuclear ERK, which was required for CPT-induced apoptosis. P38 and JNK activity were unaffected by CPT, suggesting that the effects of CPT are mediated specifically by ERK. These results suggest that targeting dual-specificity MAPK phosphatases in colon cancer cells may be a viable strategy for optimizing camptothecin-based therapeutic protocols.
Related JoVE Video
Creating dynamic images of short-lived dopamine fluctuations with lp-ntPET: dopamine movies of cigarette smoking.
J Vis Exp
PUBLISHED: 08-22-2013
Show Abstract
Hide Abstract
We describe experimental and statistical steps for creating dopamine movies of the brain from dynamic PET data. The movies represent minute-to-minute fluctuations of dopamine induced by smoking a cigarette. The smoker is imaged during a natural smoking experience while other possible confounding effects (such as head motion, expectation, novelty, or aversion to smoking repeatedly) are minimized. We present the details of our unique analysis. Conventional methods for PET analysis estimate time-invariant kinetic model parameters which cannot capture short-term fluctuations in neurotransmitter release. Our analysis--yielding a dopamine movie--is based on our work with kinetic models and other decomposition techniques that allow for time-varying parameters. This aspect of the analysis--temporal-variation--is key to our work. Because our model is also linear in parameters, it is practical, computationally, to apply at the voxel level. The analysis technique is comprised of five main steps: pre-processing, modeling, statistical comparison, masking and visualization. Preprocessing is applied to the PET data with a unique HYPR spatial filter that reduces spatial noise but preserves critical temporal information. Modeling identifies the time-varying function that best describes the dopamine effect on 11C-raclopride uptake. The statistical step compares the fit of our (lp-ntPET) model to a conventional model. Masking restricts treatment to those voxels best described by the new model. Visualization maps the dopamine function at each voxel to a color scale and produces a dopamine movie. Interim results and sample dopamine movies of cigarette smoking are presented.
Related JoVE Video
Relationship between pregnancy rate and serum progesterone concentration in porcine embryo transfer.
J. Vet. Sci.
PUBLISHED: 08-12-2013
Show Abstract
Hide Abstract
Progesterone (P4) plays a key role in implantation and pregnancy. Thus, the level of P4 at the time of embryo transfer (ET) is important. P4 concentrations and numbers of corpus lutea were evaluated from 126 surrogate pigs. SCNT embryos were transferred into 126 surrogates, among them, 11 surrogates maintained pregnancy to full-term delivery, 17 miscarried and 98 failed to establish implantation. P4 in the full-term group was significantly different from aborted pregnancy group and failed implantation group (p<0.05). However, the numbers of CLs were not significantly different. These findings indicate the concentration of progesterone can be an important factor to consider in successful ET experiments in pigs.
Related JoVE Video
Determination of the in vivo selectivity of a new ?-opioid receptor antagonist PET tracer 11C-LY2795050 in the rhesus monkey.
J. Nucl. Med.
PUBLISHED: 08-05-2013
Show Abstract
Hide Abstract
(11)C-LY2795050 is a novel ?-selective antagonist PET tracer. The in vitro binding affinities (Ki) of LY2795050 at the ?-opioid (KOR) and ?-opioid (MOR) receptors are 0.72 and 25.8 nM, respectively. Thus, the in vitro KOR/MOR binding selectivity is about 36:1. Our goal in this study was to determine the in vivo selectivity of this new KOR antagonist tracer in the monkey.
Related JoVE Video
Tiam-1, a GEF for Rac1, plays a critical role in metformin-mediated glucose uptake in C2C12 cells.
Cell. Signal.
PUBLISHED: 07-22-2013
Show Abstract
Hide Abstract
Metformin is known to stimulate glucose uptake, but the mechanism for this action is not fully understood. In this study, AMPK activators (AICAR and metformin) increased the expression of T-lymphoma invasion and metastasis-inducing protein-1 (Tiam-1), a Rac1 specific guanine nucleotide exchange factor (GEF), mRNA and protein in skeletal muscle C2C12 cells. Metformin increases the serine-phosphorylation of Tiam-1 by AMPK and induces interaction between Tiam-1 and 14-3-3. Pharmacologic inhibition of AMPK blocks this interaction, indicating that 14-3-3 may be required for induction of Tiam-1 by AMPK. Metformin also increases the phosphorylation of p21-activated kinase 1 (PAK1), a direct downstream target of Rac1, dependent on AMPK. Tiam-1 is down-regulated at high glucose concentrations in cultured cells and in the db/db mouse model of hyperglycemia. Furthermore, Tiam-1 knock-down blocked metformin-induced increase in glucose uptake. These findings suggest that metformin promotes cellular glucose uptake in part through Tiam-1 induction.
Related JoVE Video
Changes in histological features of nasal polyps in a Korean population over a 17-year period.
Otolaryngol Head Neck Surg
PUBLISHED: 06-28-2013
Show Abstract
Hide Abstract
Nasal polyps can be categorized as eosinophilic or non-eosinophilic, depending on inflammatory cell infiltration. There are geographical differences in the prevalence of types of pathologic polyps. The aim of this study was to evaluate the change in the prevalence of histological subtypes of polyps over time in a Korean population.
Related JoVE Video
Functional and molecular changes of the bladder in rats with crushing injury of nerve bundles from major pelvic ganglion to the bladder: role of RhoA/Rho kinase pathway.
Int J Mol Sci
PUBLISHED: 06-08-2013
Show Abstract
Hide Abstract
Voiding dysfunction is a common complication after radical pelvic surgery. To reduce this complication, nerve-sparing radical pelvic surgery was introduced. However, several patients experienced voiding difficulty despite nerve-sparing radical pelvic surgery. Thus, we investigated the functional and molecular changes of the bladder in rats, which demonstrated voiding dysfunction induced by nerve damage during nerve-sparing radical pelvic surgery. Male rats were used and assigned to normal, sham-operated, and bilateral crushing nerve bundles from major pelvic ganglion (MPG) to bladder group. After one, two, and four-week crushing injury, significantly decreased contractile response and increased connective tissue of the detrusor were observed and these results were reliable findings with voiding difficulty following nerve-sparing radical pelvic surgery. After crushing injury, significantly increased M2 muscarinic receptor expression was observed and this might be regarded as the compensatory response. However, M3 muscarinic receptor expression was not significantly changed. The expression of RhoA, ROCK-?, and ROCK-? was significantly increased after one, two, and four-week crushing injury. From these results, the down-regulation of RhoA/Rho kinase pathway might lead to the decreased bladder contractility after crushing injury of nerve bundles from MPG to the bladder despite of the compensated up-regulation of M2 muscarinic receptor.
Related JoVE Video
Effect of Lutein on L-NAME-Induced Hypertensive Rats.
Korean J. Physiol. Pharmacol.
PUBLISHED: 05-06-2013
Show Abstract
Hide Abstract
We investigated the antihypertensive effect of lutein on N(G) -nitro-L-arginine methyl ester hydrochloride (L-NAME)-induced hypertensive rats. Daily oral administration of L-NAME (40 mg/kg)-induced a rapid progressive increase in mean arterial pressure (MAP). L-NAME significantly increased MAP from the first week compared to that in the control and reached 193.3±9.6 mmHg at the end of treatment. MAP in the lutein groups was dose-dependently lower than that in the L-NAME group. Similar results were observed for systolic and diastolic blood pressure of L-NAME-induced hypertensive rats. The control group showed little change in heart rate for 3 weeks, whereas L-NAME significantly reduced heart rate from 434±26 to 376±33 beats/min. Lutein (2 mg/kg) significantly prevented the reduced heart rate induced by L-NAME. L-NAME caused hypertrophy of heart and kidney, and increased plasma lipid peroxidation four-fold but significantly reduced plasma nitrite and glutathione concentrations, which were significantly prevented by lutein in a dose-dependent manner. These findings suggest that lutein affords significant antihypertensive and antioxidant effects against L-NAME-induced hypertension in rats.
Related JoVE Video
Improved performance of GaN-based vertical light emitting diodes with conducting and transparent single-walled carbon nanotube networks.
Opt Express
PUBLISHED: 04-11-2013
Show Abstract
Hide Abstract
In this study, reduced forward voltage and improved light output power of GaN-based vertical light-emitting diodes (VLEDs) incorporating single-walled carbon nanotube (SWNT)-networks is reported. The SWNT-networks were directly formed on a roughened (textured) n-GaN surface via a solution-processed dip-coating method. The surface-roughened VLEDs with the proposed SWNT-networks had a forward voltage of 3.84 V at 350 mA, lower than that of the surface-roughened VLEDs, and exhibited an increase in light output power by 12.9% at 350 mA compared to the surface-roughened VLEDs. These improved electrical and optical properties could be attributed to the SWNT-networks put on the roughened n-GaN surface, which increase the lateral current transport and create scattering of light through the formation of additional roughness.
Related JoVE Video
Improvement of CNS defects via continuous intrathecal enzyme replacement by osmotic pump in mucopolysaccharidosis type II mice.
Am. J. Med. Genet. A
PUBLISHED: 03-25-2013
Show Abstract
Hide Abstract
Mucopolysaccharidosis type II (MPS II), also known as Hunter syndrome (OMIM 309900), is a rare, X-linked lysosomal storage disorder caused by a deficiency of iduronate-2-sulfatase (IDS; EC 3.1.6.13), which is involved in the lysosomal degradation of glycosaminoglycans (GAG). Although intermittent intrathecal (IT) injection of the enzyme has been introduced as a method to overcome the blood-brain barrier, continuous IT infusion of the enzyme would be more physiologic. This study was performed to investigate responses in the brain of MPS II mice to varying doses of continuous IT infusion of recombinant human IDS (rh-IDS) in MPS II mice by osmotic pump in three different doses (2.4, 4.8, and 12 µg/day) of rh-IDS for 3 weeks. The results showed that the group treated with 12 µg/day doses of rh-IDS demonstrated decreased GAG concentrations compared to the untreated KO mice group (P = 0.003). After 3 weeks of continuous IT ERT, the brain tissues of the high-dose IT-treated KO mice showed a reduction of vacuolation in the cerebral cortex, thalamus and cerebellar cortex, which was not observed in the low- and medium-dose KO mice groups. Moreover, the anti-NeuN signal representing intact neuron was restored in the cortexes of the high-dose group. In conclusion, continuous IT infusion of the deficient enzyme was effective in improving CNS defects in the MPS II mice, and could be a valuable therapeutic method for treating neurological deterioration in patients with MPS II.
Related JoVE Video
Phase I/II clinical trial of enzyme replacement therapy with idursulfase beta in patients with mucopolysaccharidosis II (Hunter syndrome).
Orphanet J Rare Dis
PUBLISHED: 03-09-2013
Show Abstract
Hide Abstract
Mucopolysaccharidosis II (MPS II, Hunter syndrome) is a rare X-linked lysosomal storage disorder caused by the deficiency of iduronate-2-sulfatase (IDS). In affected patients, glycosaminoglycan (GAG) accumulates in the lysosomes of many organs and tissues contributing to the pathology associated with MPS II. The objective of this phase I/II clinical study was to evaluate the efficacy and safety of recombinant human iduronate-2-sulfatase (idursulfase beta, Hunterase®) in the treatment of MPS II.
Related JoVE Video
Replacement of glutamine with the dipeptide derivative alanyl-glutamine enhances in vitro maturation of porcine oocytes and development of embryos.
Zygote
PUBLISHED: 02-27-2013
Show Abstract
Hide Abstract
Summary The presence of glutamine (Gln) in in vitro maturation (IVM) and in vitro culture (IVC) medium is a more potent factor for improving porcine oocyte and embryo development than other amino acids. However Gln is inherently unstable and spontaneously breaks down into ammonia, and therefore interferes with proper development. To avoid this adverse effect, Gln was replaced in the present study with its stable dipeptide derivative alanyl-glutamine (Ala-Gln) and the effects of this replacement on porcine IVM and IVC were evaluated. Replacement of Gln with Ala-Gln during IVM did not improve nuclear maturation, however numbers of early cleaved embryos were significantly increased after activation. Blastocyst formation rates were also significantly improved by using Ala-Gln during IVM. Replacement of Gln with Ala-Gln during IVC significantly increased total cell numbers in blastocysts. Blastocyst formation rate was also significantly higher when Ala-Gln was used in both IVM and IVC. In conclusion, the use of Ala-Gln rather than Gln gives better results for development in both porcine IVM and IVC.
Related JoVE Video
The effect of acteoside on intracellular Ca(2+) mobilization and phospholipase C activity in RBL-2H3 cells stimulated by melittin.
Arch. Pharm. Res.
PUBLISHED: 02-26-2013
Show Abstract
Hide Abstract
This study was performed to investigate the effects of acteoside on various cellular functions such as, intracellular Ca(2+) mobilization, phospholipase C activity, and exocytosis induced by melittin. Melittin (0.1-1 ?M) dose-dependently increased intracellular Ca(2+) mobilization in the presence of extracellular Ca(2+), but was not affected by 1 ?M U73122, a specific PLC inhibitor. In the absence of extracellular Ca(2+), melittin (1 ?M) did not induce a change in intracellular Ca(2+) mobilization, which suggests that melittin-induced intracellular Ca(2+) mobilization may be dependent on the influx of extracellular Ca(2+) rather than on the release of intracellular Ca(2+) storage. Acteoside (10 ?M) significantly inhibited 1 ?M melittin-induced Ca(2+) mobilization by 33 %. In [(3)H]inositol-labeled cells, 1 ?M melittin did not increase inositol phosphate formation, but more than 5 ?M melittin significantly increased inositol phosphate formation, which was significantly inhibited by acteoside. Melittin (1 ?M) significantly increased histamine release from RBL 2H3 cells in the presence or absence of extracellular Ca(2+). Acteoside significantly inhibited 1-?M-melittin-induced histamine release by 74 % in the presence of extracellular Ca(2+) and by 71 % in the absence of extracellular Ca(2+). These data suggest that the inhibitory effect of acteoside on 1 ?M-melittin-induced histamine release may be related to blockage of the calcium-independent pathway. Taken together, these data suggest that melittin has an influence on cellular functions such as intracellular Ca(2+) mobilization, the PLC pathway, and exocytosis via various independent signalling pathways in RBL-2H3 cells, and was significantly inhibited by acteoside.
Related JoVE Video
Influence of type of nocturia and lower urinary tract symptoms on therapeutic outcome in women treated with desmopressin.
Korean J Urol
PUBLISHED: 02-18-2013
Show Abstract
Hide Abstract
To investigate the type of nocturia and concomitant voiding dysfunction (VD) and the effect of desmopressin treatment on nocturia in women.
Related JoVE Video
Osteogenesis imperfecta Type VI with severe bony deformities caused by novel compound heterozygous mutations in SERPINF1.
J. Korean Med. Sci.
PUBLISHED: 02-15-2013
Show Abstract
Hide Abstract
Osteogenesis imperfecta (OI) comprises a heterogeneous group of disorders characterized by bone fragility, frequent fractures, and low bone mass. Dominantly inherited COL1A1 or COL1A2 mutations appear to be causative in the majority of OI types, but rare recessively inherited genes have also been reported. Recently, SERPINF1 has been reported as another causative gene in OI type VI. To date, only eight SERPINF1 mutations have been reported and all are homozygous. Our patient showed no abnormalities at birth, frequent fractures, osteopenia, and poor response on pamidronate therapy. At the time of her most recent evaluation, she was 8 yr old, and could not walk independently due to frequent lower-extremity fractures, resulting in severe deformity. No clinical signs were seen of hearing impairment, blue sclera, or dentinogenesis imperfecta. In this study, we describe the clinical and radiological findings of one Korean patient with novel compound heterozygous mutations (c.77dupC and c.421dupC) of SERPINF1.
Related JoVE Video
Efficacy and safety of low-dose anticholinergics to treat men with lower urinary tract symptoms with overactive bladder: a retrospective study based on real life practice.
Prostate Int
PUBLISHED: 02-02-2013
Show Abstract
Hide Abstract
To investigate whether combination treatment using an ?-blocker and 2 mg of tolterodine could improve the International Prostate Symptom Score (IPSS) as much as ?-blocker and 4 mg of tolterodine without voiding difficulties in real life practice.
Related JoVE Video
Synthesis and evaluation of 11C-LY2795050 as a ?-opioid receptor antagonist radiotracer for PET imaging.
J. Nucl. Med.
PUBLISHED: 01-25-2013
Show Abstract
Hide Abstract
Kappa-opioid receptors (KOR) are believed to be involved in the pathophysiology of depression, anxiety disorders, drug abuse, and alcoholism. To date, only 1 tracer, the KOR agonist (11)C-GR103545, has been reported to be able to image KOR in primates. The goal of the present study was to synthesize the selective KOR antagonist (11)C-LY2795050 and evaluate its potential as a PET tracer to image KOR in vivo.
Related JoVE Video
Risk Factors for Failure of Early Catheter Removal After Greenlight HPS Laser Photoselective Vaporization Prostatectomy in Men With Benign Prostatic Hyperplasia.
Korean J Urol
PUBLISHED: 01-18-2013
Show Abstract
Hide Abstract
To assess the risk factors for developing urinary retention after removal of the urethral catheter on postoperative day 1 in benign prostatic hyperplasia patients who underwent Greenlight HPS laser photoselective vaporization prostatectomy (PVP).
Related JoVE Video
Digging the New York City Skyline: soil fungal communities in green roofs and city parks.
PLoS ONE
PUBLISHED: 01-15-2013
Show Abstract
Hide Abstract
In urban environments, green roofs provide a number of benefits, including decreased urban heat island effects and reduced energy costs for buildings. However, little research has been done on the non-plant biota associated with green roofs, which likely affect their functionality. For the current study, we evaluated whether or not green roofs planted with two native plant communities in New York City functioned as habitats for soil fungal communities, and compared fungal communities in green roof growing media to soil microbial composition in five city parks, including Central Park and the High Line. Ten replicate roofs were sampled one year after planting; three of these roofs were more intensively sampled and compared to nearby city parks. Using Illumina sequencing of the fungal ITS region we found that green roofs supported a diverse fungal community, with numerous taxa belonging to fungal groups capable of surviving in disturbed and polluted habitats. Across roofs, there was significant biogeographical clustering of fungal communities, indicating that community assembly of roof microbes across the greater New York City area is locally variable. Green roof fungal communities were compositionally distinct from city parks and only 54% of the green roof taxa were also found in the park soils. Phospholipid fatty acid analysis revealed that park soils had greater microbial biomass and higher bacterial to fungal ratios than green roof substrates. City park soils were also more enriched with heavy metals, had lower pH, and lower quantities of total bases (Ca, K, and Mg) compared to green roof substrates. While fungal communities were compositionally distinct across green roofs, they did not differentiate by plant community. Together, these results suggest that fungi living in the growing medium of green roofs may be an underestimated component of these biotic systems functioning to support some of the valued ecological services of green roofs.
Related JoVE Video
The proportion of uniparental disomy is increased in Prader-Willi syndrome due to an advanced maternal childbearing age in Korea.
J. Hum. Genet.
PUBLISHED: 01-10-2013
Show Abstract
Hide Abstract
Prader-Willi syndrome (PWS) is a genetic disorder caused by the absence of expression of the paternal copy of maternally imprinted genes in chromosome region 15q11-13. The genetic subtypes of PWS are classified into deletion (~70%), maternal uniparental disomy (mUPD; 25-30%), imprinting center defects (3-5%) and rare unbalanced translocations. Recently, Matsubara et al. reported a significantly higher maternal age in a trisomy rescue (TR) or gamete complementation (GC) by nondisjunction at maternal meiosis 1 (M1) group than in a deletion group. In the present study, we try to confirm their findings in an ethnically different population. A total of 97 Korean PWS patients were classified into deletional type (n=66), TR/GC (M1) (n=15), TR/GC by nondisjunction at maternal meiosis 2 (n=2), monosomy rescue or postfertilization mitotic nondisjunction (n=4) and epimutation (n=2). Maternal ages at birth showed a significant difference between the deletion group (median age of 29, interquartile range (IQR)=(27,31)) and the TR/GC (M1) group (median age of 35, IQR=(31,38)) (P<0.0001). The relative birth frequency of the TR/GC (M1) group has substantially increased since 2006 when compared with the period before 2005. These findings support the hypothesis that the advanced maternal age at childbirth is a predisposing factor for the development of mUPD because of increased M1 errors.
Related JoVE Video
The influence of ethanol intake on RhoA/Rho kinase signaling pathway in corpus cavernosum of OLETF rats.
Int Urol Nephrol
PUBLISHED: 01-05-2013
Show Abstract
Hide Abstract
To investigate the effect of the ethanol on corpus cavernosum of the type 2 diabetes mellitus (type 2 DM) rat, since previous studies have reported the positive effect of moderate drinking on type 2 DM and endothelial function.
Related JoVE Video
Quantitative assessment of partial vascular occlusions in a swine pedicle flap model using spatial frequency domain imaging.
Biomed Opt Express
PUBLISHED: 01-04-2013
Show Abstract
Hide Abstract
The use of tissue transfer flaps has become a common and effective technique for reconstructing or replacing damaged tissue. While the overall failure rate associated with these procedures is relatively low (5-10%), the failure rate of tissue flaps that require additional surgery is significantly higher (40-60%). The reason for this is largely due to the absence of a technique for objectively assessing tissue health after surgery. Here we have investigated spatial frequency domain imaging (SFDI) as a potential tool to do this. By projecting wide-field patterned illumination at multiple wavelengths onto a tissue surface, SFDI is able to quantify absolute concentrations of oxygenated and deoxygenated hemoglobin over a large field of view. We have assessed the sensitivity of SFDI in a swine pedicle flap model by using a controlled vascular occlusion system that reduced blood flow by 25%, 50%, 75%, or 100% of the baseline values in either the vein or artery. SFDI was able to detect significant changes for oxygenated hemoglobin, deoxygenated hemoglobin, or tissue oxygen saturation in partial arterial occlusions of at least 50% and partial venous occlusions of at least 25%. This shows SFDI is sensitive enough to quantify changes in the tissue hemoglobin state during partial occlusions and thus has the potential to be a powerful tool for the early prediction of tissue flap failure.
Related JoVE Video
E3 ubiquitin ligase, WWP1, interacts with AMPK?2 and down-regulates its expression in skeletal muscle C2C12 cells.
J. Biol. Chem.
PUBLISHED: 01-04-2013
Show Abstract
Hide Abstract
It is known that the activity of AMP-activated protein kinase (AMPK?2) was depressed under high glucose conditions. However, whether protein expression of AMPK?2 is also down-regulated or not remains unclear. In this study, we showed that the expression of AMPK?2 was down-regulated in cells cultured under high glucose conditions. Treatment of proteasome inhibitor, MG132, blocked high glucose-induced AMPK?2 down-regulation. Endogenous AMPK?2 ubiquitination was detected by immunoprecipitation of AMPK?2 followed by immunoblotting detection of ubiquitin. The yeast-two hybrid (YTH) approach identified WWP1, an E3 ubiquitin ligase, as the AMPK?2-interacting protein in skeletal muscle cells. Interaction between AMPK?2 and WWP1 was validated by co-immunoprecipitation. Knockdown of WWP1 blocked high glucose-induced AMPK?2 down-regulation. The overexpression of WWP1 down-regulated AMPK?2. In addition, the expression of WWP1 is increased under high glucose culture conditions in both mRNA and protein levels. The level of AMPK?2 was down-regulated in the quadriceps muscle of diabetic animal model db/db mice. Expression of WWP1 blocked metformin-induced glucose uptake. Taken together, our results demonstrated that WWP1 down-regulated AMPK?2 under high glucose culture conditions via the ubiquitin-proteasome pathway.
Related JoVE Video
Advantages of Single-Molecule Real-Time Sequencing in High-GC Content Genomes.
PLoS ONE
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
Next-generation sequencing has become the most widely used sequencing technology in genomics research, but it has inherent drawbacks when dealing with high-GC content genomes. Recently, single-molecule real-time sequencing technology (SMRT) was introduced as a third-generation sequencing strategy to compensate for this drawback. Here, we report that the unbiased and longer read length of SMRT sequencing markedly improved genome assembly with high GC content via gap filling and repeat resolution.
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.