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Find video protocols related to scientific articles indexed in Pubmed.
Localized Electric Field of Plasmonic Nanoplatform Enhanced Photodynamic Tumor Therapy.
ACS Nano
PUBLISHED: 11-07-2014
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Near-infrared plasmonic nanoparticles demonstrate great potential in disease theranostic applications. Herein a nanoplatform, composed of mesoporous silica-coated gold nanorods (AuNRs), is tailor-designed to optimize the photodynamic therapy (PDT) for tumor based on the plasmonic effect. The surface plasmon resonance of AuNRs was fine-tuned to overlap with the exciton absorption of indocyanine green (ICG), a near-infrared photodynamic dye with poor photostability and low quantum yield. Such overlap greatly increases the singlet oxygen yield of incorporated ICG by maximizing the local field enhancement, and protecting the ICG molecules against photodegradation by virtue of the high absorption cross section of the AuNRs. The silica shell strongly increased ICG payload with the additional benefit of enhancing ICG photostability by facilitating the formation of ICG aggregates. As-fabricated AuNR@SiO2-ICG nanoplatform enables trimodal imaging, near-infrared fluorescence from ICG, and two-photon luminescence/photoacoustic tomography from the AuNRs. The integrated strategy significantly improved photodynamic destruction of breast tumor cells and inhibited the growth of orthotopic breast tumors in mice, with mild laser irradiation, through a synergistic effect of PDT and photothermal therapy. Our study highlights the effect of local field enhancement in PDT and demonstrates the importance of systematic design of nanoplatform to greatly enhancing the antitumor efficacy.
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Efficient Generation of Marker-Free Transgenic Rice Plants Using an Improved Transposon-Mediated Transgene Reintegration Strategy.
Plant Physiol.
PUBLISHED: 11-04-2014
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Marker-free transgenic plants can be developed through transposon-mediated transgene reintegration, which allows intact transgene insertion with defined boundaries and requires only a few primary transformants. In this study, we improved the selection strategy and validated that the maize Ac/Ds transposable element can be routinely used to generate marker-free transgenic plants. A Ds-based gene of interest (GOI) was linked to green fluorescent protein (GFP) in T-DNA, and a GFP-aided counter-selection against T-DNA was used together with PCR-based positive selection for the GOI to screen marker-free progeny. To test the efficacy of this strategy, we cloned the Bacillus thuringiensis (Bt) ð-endotoxin gene into the Ds elements and transformed transposon vectors into rice cultivars via Agrobacterium. PCR assays of the transposon empty donor site exhibited transposition in somatic cells in 60.5% to 100% of the rice transformants. Marker-free (T-DNA-free) transgenic rice plants derived from unlinked germinal transposition were obtained from the T1 generation of 26.1% of the primary transformants. Individual marker-free transgenic rice lines were subjected to TAIL-PCR to determine Ds(Bt) reintegration positions, RT-PCR and ELISA to detect Bt expression levels, and bioassays to confirm resistance against the striped stem borer Chilo suppressalis (Walker). Overall, we efficiently generated marker-free transgenic plants with optimized transgene insertion and expression. The transposon-mediated marker-free platform established in this study can be used in rice and possibly in other important crops.
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Morphology-Controlled Self-Assembly and Synthesis of Photocatalytic Nanocrystals.
Nano Lett.
PUBLISHED: 11-04-2014
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Abilities to control the size and shape of nanocrystals in order to tune functional properties are an important grand challenge. Here we report a surfactant self-assembly induced micelle encapsulation method to fabricate porphyrin nanocrystals using the optically active precursor zinc porphyrin (ZnTPP). Through confined noncovalent interactions of ZnTPP within surfactant micelles, nanocrystals with a series of morphologies including nanodisk, tetragonal rod, and hexagonal rod, as well as amorphous spherical particle are synthesized with controlled size and dimension. A phase diagram that describes morphology control is achieved via kinetically controlled nucleation and growth. Because of the spatial ordering of ZnTPP, the hierarchical nanocrystals exhibit both collective optical properties resulted from coupling of molecular ZnTPP and shape dependent photocatalytic activities in photo degradation of methyl orange pollutants. This simple ability to exert rational control over dimension and morphology provides new opportunities for practical applications in photocatalysis, sensing, and nanoelectronics.
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[Potential risk factors of excessive epistaxis after endoscopic endonasal surgery].
Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi
PUBLISHED: 10-22-2014
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To investigate the potential risk factors and management of excessive epistaxis after endoscopic endonasal surgery (EES).
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Rapid identification of synthetic colorants in food samples by using indium oxide nanoparticle-functionalized porous polymer monolith coupled with HPLC-MS/MS.
Analyst
PUBLISHED: 10-15-2014
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A synthetic protocol for the preparation of an indium oxide nanoparticle-functionalized poly(methacrylic acid-glycidyl methacrylate-ethylene dimethacrylate-ethanediamine) monolithic column is reported. Various techniques, including scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, and thermal gravimetric analysis-derivative thermogravimetric analysis were employed to characterize the synthesized monolith. The modified monolithic column was coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) for determining synthetic colorants in various food samples. Under optimized conditions, good linearity was obtained for all the targets with squared regression coefficients greater than 0.9982. The limits of detection (S/N = 3) for 12 synthetic colorants were in the range of 0.012-2.97 ?g kg(-1). The intra-day and inter-day relative standard deviations, ranging from 2.7% to 8.5%, were within the acceptable range. The developed method was successfully applied to the determination of synthetic colorants in food samples (candy, milk, jelly, jam, canned food, juice, and carbonated drink). Target recoveries at different spiked levels ranged from 73.5% to 112.1% with relative standard deviations of less than 10.3%.
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Transcriptional profiling reveals crosstalk between mesenchymal stem cells and endothelial cells promoting pre-vascularization by reciprocal mechanisms.
Stem Cells Dev.
PUBLISHED: 10-10-2014
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Mesenchymal stem cells (MSCs) show great promise in blood vessel restoration and vascularization enhancement in many therapeutic situations. Typically, the co-implantation of MSCs with vascular endothelial cells (ECs) is effective for the induction of functional vascularization in vivo, indicating its potential applications in regenerative medicine. The effects of MSCs-ECs-induced vascularization can be modeled in vitro, providing simplified models for understanding their underlying communication. In this paper, a contact co-culture model in vitro and an RNA-seq approach were employed to reveal the active crosstalk between MSCs and ECs within a short time period at both morphological and transcriptional levels. The RNA-seq results suggested that angiogenic genes were significantly induced upon co-culture, and this pre-vascularization commitment might require the NF-?B signaling. NF-?B blocking and interleukin neutralization experiments demonstrated that MSCs potentially secreted interleukin factors including IL1? and IL6 to modulate NF-?B signaling as well as downstream chemokines during co-culture. Conversely, RNA-seq results indicated that the MSCs were regulated by the co-culture environment to a smooth muscle commitment within this short period, which largely induced myocardin, the myogenic co-transcriptional factor. These findings demonstrate the mutual molecular mechanism of MSCs-ECs-induced pre-vascularization commitment in a quick response.
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[Application of different technologies for distinguishing true and pseudo mosaicisms during prenatal diagnosis].
Zhonghua Yi Xue Yi Chuan Xue Za Zhi
PUBLISHED: 10-10-2014
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To use different technologies to distinguish true and pseudo mosaicisms among cultured amniocytes in order to attain more accurate diagnosis.
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Incorporation of graphene oxide nanosheets into boronate-functionalized polymeric monolith to enhance the electrochromatographic separation of small molecules.
Electrophoresis
PUBLISHED: 09-28-2014
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Graphene oxide (GO) nanosheets were incorporated into an organic polymer monolith containing 3-acrylamidophenylboronic acid (AAPBA) and pentaerythritol triacrylate (PETA) to form a novel monolithic stationary phase for capillary electrochromatohraphy (CEC). The effects of the mass ratio of AAPBA/PETA, the amount of GO, and the volume of porogen on the morphology, permeability and pore properties of the prepared poly (AAPBA-GO-PETA) monoliths were investigated. A series of test compounds including amides, alkylbenzenes, polycyclic aromatics, phenols and anilines were used to evaluate and compare the separation performances of the poly (AAPBA-GO-PETA) and the parent poly (AAPBA-co-PETA) monoliths. The results indicated that incorporation of GO into monolithic oclumn exhibited much higher resolutions (>1.5) and column efficiency (62 000?110 000 plates/m for toluene, DMF, formamide and thiourea) than the poly (AAPBA-co-PETA). The successful application in isocratic separation of peptides suggests the potential of the GO incorporated monolithic column in complex sample analysis. In addition, the reproducibility and stability of the prepared poly (AAPBA-GO-PETA) monolith was assessed. The run-to-run, column-to-column and batch-to-batch reproducibilities of this monolith for alkylbenzenes' retention were satisfactory with the relative standard deviations (RSDs) less than 1.8% (n = 5), 3.7% and 5.6% (n = 3), respectively, indicating the effectiveness and practicability of the proposed method. This article is protected by copyright. All rights reserved.
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[Characterization of a recombinant goatpox virus expressing Orfv F1L gene].
Wei Sheng Wu Xue Bao
PUBLISHED: 09-26-2014
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In order to establish the vaccine against the contagious ecthyma, we constructed and characterized recombinant goatpox virus expressing F1L protein of Orf virus.
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hNAG-1 increases lifespan by regulating energy metabolism and insulin/IGF-1/mTOR signaling.
Aging (Albany NY)
PUBLISHED: 09-21-2014
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Nonsteroidal anti-inflammatory drug-activated gene (NAG-1) or GDF15 is a divergent member of the transforming growth factor beta (TGF-?) superfamily and mice expressing hNAG-1/hGDF15 have been shown to be resistant to HFD-induced obesity and inflammation. This study investigated if hNAG-1 increases lifespan in mice and its potential mechanisms. Here we report that female hNAG-1 mice had significantly increased both mean and median life spans in two transgenic lines, with a larger difference in life spans in mice on a HFD than on low fat diet. hNAG-1 mice displayed significantly reduced body and adipose tissue weight, lowered serum IGF-1, insulin and glucose levels, improved insulin sensitivity, and increased oxygen utilization, oxidative metabolism and energy expenditure. Gene expression analysis revealed significant differences in conserved gene pathways that are important regulators of longevity, including IGF-1, p70S6K, and PI3K/Akt signaling cascades. Phosphorylation of major components of IGF-1/mTOR signaling pathway was significantly lower in hNAG-1mice. Collectively, hNAG-1 is an important regulator of mammalian longevity and may act as a survival factor. Our study suggests that hNAG-1 has potential therapeutic uses in obesity-related diseases where life span is frequently shorter.
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Role of miR-195 in Aortic Aneurysmal Disease.
Circ. Res.
PUBLISHED: 09-08-2014
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Abdominal aortic aneurysms constitute a degenerative process in the aortic wall. Both the miR-29 and miR-15 families have been implicated in regulating the vascular extracellular matrix.
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RDH5 RETINOPATHY (FUNDUS ALBIPUNCTATUS) WITH PRESERVED ROD FUNCTION IN YOUNG CHINESE PATIENT.
Retina (Philadelphia, Pa.)
PUBLISHED: 08-28-2014
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The aim of this study is to characterize the clinical features of four unrelated Chinese patients with retinal dehydrogenase 5 (RDH5) retinopathy (fundus albipunctatus) and to identify the genetic defects underlying this disorder.
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Model Drug as Pore Former for Controlled Release of Water-Soluble Metoprolol Succinate from Ethylcellulose-Coated Pellets Without Lag Phase: Opportunities and Challenges.
AAPS PharmSciTech
PUBLISHED: 08-28-2014
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The objective of the present study was to evaluate the feasibility of using model drug metoprolol succinate (MS) as a pore former to modify the initial lag phase (i.e., a slow or non-release phase in the first 1-2 h) associated with the drug release from coated pellets. MS-layered cores with high drug-layering efficiency (97% w/w) were first prepared by spraying a highly concentrated drug aqueous solution (60% w/w, 70°C) on non-pareils without using other binders. The presence of MS in ethylcellulose (EC) coating solution significantly improved the coating process by reducing pellets sticking, which often occurs during organic coating. There may be a maximum physical compatibility of MS with EC, and the physical state of the drug in the functional coating layer of EC/MS (80:20) was simultaneously crystalline and non-crystalline (amorphous or solid molecule solution). The lag phase associated with hydroxypropylcellulose (HPC) as a pore former was not observed when MS was used as a pore former. The drug release from EC/MS-coated pellets was pH independent, inversely proportional to the coating levels, and directly related to the pore former levels. The functional coating layer with MS as a pore former was not completely stabilized without curing. Curing at 60°C for 1 day could substantially improve the stability of EC/MS-coated pellets. The physical state of the drug in the free film of EC/MS (85:15) changed partially from amorphous to crystal when cured at 60°C for 1 day, which should be attributed to the incompatibility of the drug with EC.
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Quantifying the effect of trend, fluctuation, and extreme event of climate change on ecosystem productivity.
Environ Monit Assess
PUBLISHED: 08-15-2014
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Climate change comprises three fractions of trend, fluctuation, and extreme event. Assessing the effect of climate change on terrestrial ecosystem requires an understanding of the action mechanism of these fractions, respectively. This study examined 11 years of remotely sensed-derived net primary productivity (NPP) to identify the impacts of the trend and fluctuation of climate change as well as extremely low temperatures caused by a freezing disaster on ecosystem productivity in Hunan province, China. The partial least squares regression model was used to evaluate the contributions of temperature, precipitation, and photosynthetically active radiation (PAR) to NPP variation. A climatic signal decomposition and contribution assessment model was proposed to decompose climate factors into trend and fluctuation components. Then, we quantitatively evaluated the contributions of each component of climatic factors to NPP variation. The results indicated that the total contribution of the temperature, precipitation, and PAR to NPP variation from 2001 to 2011 in Hunan province is 85 %, and individual contributions of the temperature, precipitation, and PAR to NPP variation are 44 % (including 34 % trend contribution and 10 % fluctuation contribution), 5 % (including 4 % trend contribution and 1 % fluctuation contribution), and 36 % (including 30 % trend contribution and 6 % fluctuation contribution), respectively. The contributions of temperature fluctuation-driven NPP were higher in the north and lower in the south, and the contributions of precipitation trend-driven NPP and PAR fluctuation-driven NPP are higher in the west and lower in the east. As an instance of occasionally triggered disturbance in 2008, extremely low temperatures and a freezing disaster produced an abrupt decrease of NPP in forest and grass ecosystems. These results prove that the climatic trend change brought about great impacts on ecosystem productivity and that climatic fluctuations and extreme events can also alter the ecosystem succession process, even resulting in an alternative trajectory. All of these findings could improve our understanding of the impacts of climate change on the provision of ecosystem functions and services and can also provide a basis for policy makers to apply adaptive measures to overcome the unfavorable influence of climate change.
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Template-Free Synthesis of Core-Shell TiO2 Microspheres Covered with High-Energy {116}-Facet-Exposed N-Doped Nanosheets and Enhanced Photocatalytic Activity under Visible Light.
Chemistry
PUBLISHED: 08-14-2014
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Core-shell TiO2 microspheres possess a unique structure and interesting properties, and therefore, they have received much attention. The high-energy facets of TiO2 also are being widely studied for the high photocatalytic activities they are associated with. However, the synthesis of the core-shell structure is difficult to achieve and requires multiple-steps and/or is expensive. Hydrofluoric acid (HF), which is highly corrosive, is usually used in the controlling high-energy facet production. Therefore, it is still a significant challenge to develop low-temperature, template-free, shape-controlled, and relative green self-assembly routes for the formation of core-shell-structured TiO2 microspheres with high-energy facets. Here, we report a template- and hydrofluoric acid free solvothermal self-assembly approach to synthesize core-shell TiO2 microspheres covered with high-energy {116}-facet-exposed nanosheets, an approach in which 1,4-butanediamine plays a key role in the formation of nanosheets with exposed {116} facets and the doping of nitrogen in situ. In the structure, nanoparticle aggregates and nanosheets with {116} high-energy facets exposed act as core and shell, respectively. The photocatalytic activity for degradation of 2,4,6-tribromophenol and Rhodamine B under visible irradiation and UV/Vis irradiation has been examined, and improved photocatalytic activity under visible light owing to the hierarchical core-shell structure, {116}-plane-oriented nanosheets, in situ N doping, and large surface areas has been found.
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Functional characteristics of reversibly immortalized hepatic progenitor cells derived from mouse embryonic liver.
Cell. Physiol. Biochem.
PUBLISHED: 08-11-2014
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Liver is a vital organ and retains its regeneration capability throughout adulthood, which requires contributions from different cell populations, including liver precursors and intrahepatic stem cells. To overcome the mortality of hepatic progenitors (iHPs) in vitro, we aim to establish reversibly immortalized hepatic progenitor cells from mouse embryonic liver.
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Interplay between longitudinal and transverse contrasts in Fe3O4 nanoplates with (111) exposed surfaces.
ACS Nano
PUBLISHED: 08-06-2014
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Iron oxide has been developed as either T1 or T2 magnetic resonance imaging (MRI) contrast agents by controlling the size and composition; however, the underlying mechanism of T1 and T2 contrasts in one iron oxide entity is still not well understood. Herein, we report that freestanding superparamagnetic magnetite nanoplates with (111) exposed facets have significant but interactional T1 and T2 contrast effects. We demonstrate that the main contribution of the T1 contrast of magnetic nanoplates is the chemical exchange on the iron-rich Fe3O4(111) surfaces, whereas the T2 relaxation is dominated by the intrinsic superparamagnetism of the nanoplates with an enhanced perturbation effect. We are able to regulate the balance of T1 and T2 contrasts by controlling structure and surface features, including morphology, exposed facets, and surface coating. This study provides an insightful understanding on the T1 and T2 contrast mechanisms, which is urgently needed to allow more sophisticated design of high-performance MRI contrast agents.
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Oral 9-cis retinoid for childhood blindness due to Leber congenital amaurosis caused by RPE65 or LRAT mutations: an open-label phase 1b trial.
Lancet
PUBLISHED: 07-13-2014
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Leber congenital amaurosis, caused by mutations in RPE65 and LRAT, is a severe form of inherited retinal degeneration leading to blindness. We aimed to assess replacement of the missing chromophore 11-cis retinal with oral QLT091001 (synthetic 9-cis-retinyl acetate) in these patients.
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Urinary monohydroxylated metabolites of polycyclic aromatic hydrocarbons in children living in city and rural residences in Southern China.
Environ Technol
PUBLISHED: 06-23-2014
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Urinary 2-hydroxynaphthalene, 2-hydoxyfluorene, 9-hydroxyphenanthrene, 1-hydroxypyrene and 3-hydroxybenz[a]pyrene concentrations in 179 randomly selected voluntary students were determined in the Southern China, aged 14-16 and living in four areas with different levels of polycyclic aromatic hydrocarbons (PAHs) in soil, water and ambient air. The excretion of 1-hydroxypyrene is significantly higher in students of the urban than in students of the rural, while there are no significant differences of urinary 2-hydroxynaphthalene, 2-hydoxyfluorene and 9-hydroxyphenanthrene between urban and rural children. Mean concentrations of 1-hydroxypyrene (0.54-0.80 ?mol/mol creatinine) in the study are much higher than those in the children of Denmark, Germany, Spain, USA, Korea, Japan and Taiwan, and a little higher than those in the children of Ukraine and Thailand. Urinary 2-hydroxynaphthalene concentrations in the study are a little higher than those in the children of USA, and similar to that in non-occupational exposure residences in Korea. Urinary 9-hydroxyphenanthrene concentrations in China are much higher than those in the children of USA. Differences between children with smoking parents and non-smoking parents are not significant in the study.
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Expression of DNA topoisomerase II-?: Clinical significance in laryngeal carcinoma.
Oncol Lett
PUBLISHED: 06-19-2014
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DNA topoisomerase II-? (Topo II-?) is essential for numerous cell processes, including DNA replication, transcription, recombination, and chromosome separation and condensation. Altered Topo II-? expression may lead to carcinogenesis and cancer progression. The aim of the present study was to investigate the association between Topo II-? expression levels and clinicopathological data from laryngeal cancer patients. Immunohistochemistry was used to analyze Topo II-? expression in laryngeal squamous cell carcinoma and distant healthy tissues obtained from 70 patients. In addition, fluorescence in situ hybridization was used to detect Topo II-? amplification and chromosome 17 ploidy using a laryngeal cancer tissue microarray. The expression of Topo II-? protein was detected in 71.43% (50/70) of laryngeal carcinoma tissues, in contrast to 9% of healthy tissues (2/22). Furthermore, the expression of Topo II-? protein was found to be associated with tumor de-differentiation and advanced tumor T stage. However, the expression of Topo II-? protein was not identified to be associated with Topo II-? amplification in laryngeal carcinoma, although was found to positively correlate with chromosome 17 aneuploidy (P<0.05). A higher aneuploidy rate contributed to increased expression levels of Topo II-? protein. Aberrant Topo II-? expression and chromosome 17 aneuploidy contributed to the development and progression of laryngeal cancer, indicating that targeting Topo II-? may provide a treatment strategy for patients with laryngeal cancer.
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miRNA-128 suppresses prostate cancer by inhibiting BMI-1 to inhibit tumor-initiating cells.
Cancer Res.
PUBLISHED: 06-05-2014
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microRNA-128 (miR128) is reduced in prostate cancer relative to normal/benign prostate tissues, but causal roles are obscure. Here we show that exogenously introduced miR128 suppresses tumor regeneration in multiple prostate cancer xenograft models. Cancer stem-like cell (CSC)-associated properties were blocked, including holoclone and sphere formation as well as clonogenic survival. Using a miR128 sensor to distinguish cells on the basis of miR128 expression, we found that miR128-lo cells possessed higher clonal, clonogenic, and tumorigenic activities than miR128-hi cells. miR128 targets the stem cell regulatory factors BMI-1, NANOG, and TGFBR1, the expression of which we found to vary inversely with miR128 expression in prostate cancer stem/progenitor cell populations. In particular, we defined BMI-1 as a direct and functionally relevant target of miR128 in prostate cancer cells, where these genes were reciprocally expressed and exhibited opposing biological functions. Our results define a tumor suppressor function for miR128 in prostate cancer by limiting CSC properties mediated by BMI-1 and other central stem cell regulators, with potential implications for prostate cancer gene therapy.
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Constitutive overexpression of soybean plasma membrane intrinsic protein GmPIP1;6 confers salt tolerance.
BMC Plant Biol.
PUBLISHED: 05-12-2014
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Under saline conditions, plant growth is depressed via osmotic stress and salt can accumulate in leaves leading to further depression of growth due to reduced photosynthesis and gas exchange. Aquaporins are proposed to have a major role in growth of plants via their impact on root water uptake and leaf gas exchange. In this study, soybean plasma membrane intrinsic protein 1;6 (GmPIP1;6) was constitutively overexpressed to evaluate the function of GmPIP1;6 in growth regulation and salt tolerance in soybean.
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Non-invasive assessment of liver fibrosis in patients with alcoholic liver disease using acoustic radiation force impulse elastography.
Abdom Imaging
PUBLISHED: 05-10-2014
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To investigate the diagnostic performance of the acoustic radiation force impulse (ARFI) elastography for the assessment of the liver fibrosis in alcoholic liver disease (ALD).
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Dietary supplementation with fish oil alters the expression levels of proteins governing mitochondrial dynamics and prevents high-fat diet-induced endothelial dysfunction.
Br. J. Nutr.
PUBLISHED: 04-28-2014
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Diets supplemented with fish oil (FO), which is rich in n-3 PUFA, have been shown to modify several key risk factors for CVD. The purpose of the present study was to determine the effect of FO supplementation on mitochondrial dynamic protein expression in the endothelium and on endothelial cell function. Male apoE-deficient (apoE-/-) mice (8 weeks old, n 12 per group) were fed a high-fat diet containing 45% fat (HFD group) or a HFD with partial replacement of lard with 10% (w/w) FO (FO group) (total EPA and DHA content 64.1 g/kg) for 8 weeks. ApoE-/- mice in the FO group had a greater endothelium-dependent vasorelaxation response to acetylcholine (Ach) than those in the HFD group. The atherosclerotic lesion volume in the aortic sinus of mice in the FO group was 54% lower than that in the HFD group (P< 0.01). In addition, the aortas isolated from mice in the FO group had higher expression levels of Mfn2 and Opa1 but lower expression levels of Fis1 than those from the HFD group. Compared with mice fed the HFD, those fed the FO diet showed significantly lower levels of mitochondrial oxidative stress, cytochrome c release and caspase-3 activity (each P< 0.05). Furthermore, FO-fed mice displayed increased NO release and availability and enhanced endothelial NO synthase activity compared with HFD-fed mice. Taken together, these results reveal a novel mechanism by which FO protects against endothelial cell dysfunction, which may result in improved mitochondrial dynamics.
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Biological monitoring of environmental exposure to polycyclic aromatic hydrocarbons in subjects living in the area of recycling electronic garbage, in Southern China.
Environ Sci Pollut Res Int
PUBLISHED: 04-02-2014
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The study was undertaken to evaluate the environmental exposure to polycyclic aromatic hydrocarbons in subjects living in the area of recycling electronic garbage in Southern China and research the influence of environment smoke tobacco (EST) to people through active and passive smoking. Urinary concentrations of 2-hydroxynaphthalene, 2-hydoxyfluorene, 9-hydroxyphenanthrene, and 1-hydroxypyrene were determined in 141 randomly selected voluntary residents aged 13 to 81 years in two polycyclic aromatic hydrocarbon (PAH)-exposed groups, two control groups, and an EST research group. The concentrations of 2-hydroxynaphthalene, 2-hydoxyfluorene, 9-hydroxyphenanthrene, and 1-hydroxypyrene in PAH-exposed groups are significantly higher (p<0.05) than those of control groups. Mean value of 1-hydroxypyrene in the residents living in the area of recycling electronic garbage (1.1 ?mol/mol creatinine) is a little higher than those of iron foundry workers, automobile repair workers, and firefighters. Mean value of 2-hydroxynaphthalene (11.3 ?mol/mol creatinine) is much higher than that of shipyard and aircraft maintenance and much lower than some occupational exposure, such as coking batteries, sorting department, and distillation department in coking plant. Some metabolites of PAHs (PAHm) are significantly elevated through active and passive smoking, while the influence of EST to other PAHm is not statistically significant. 2-Hydroxynaphthalene, 2-hydoxyfluorene, 9-hydroxyphenanthrene, and 1-hydroxypyrene in the urine of smokers are, respectively, 3.9, 1.9, 1.4, and 1.9 times to those of nonsmokers. In nonsmokers, passive smokers excreted 1.1, 1.5, 1.9, and 1.5 times of 2-hydroxynaphthalene, 2-hydoxyfluorene, 9-hydroxyphenanthrene, and 1-hydroxypyrene compared to nonpassive smokers.
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[Multi-center study of premature thelarche and gynecomastia in Chinese infants and toddlers].
Zhonghua Er Ke Za Zhi
PUBLISHED: 04-01-2014
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The term "premature thelarche" refers to isolated breast development before 8 years of age in female, without any other signs of sexual maturation, while "gynecomastia" is the presence of breast tissue in males. This study aimed to investigate the prevalence of premature thelarche and gynecomastia in Chinese infants and toddlers, identify the potential risk factors, and explore the influence of early breast development on physical growth, mental development and psychomotor development.
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The investigation of reducing PAHs emission from coal pyrolysis by gaseous catalytic cracking.
ScientificWorldJournal
PUBLISHED: 03-28-2014
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The catalytic cracking method of PAHs for the pyrolysis gaseous products is proposed to control their pollution to the environment. In this study, the Py-GC-MS is used to investigate in situ the catalytic effect of CaO and Fe2O3 on the 16 PAHs from Pingshuo coal pyrolysis under different catalytic temperatures and catalyst particle sizes. The results demonstrate that Fe2O3 is effective than that of CaO for catalytic cracking of 16 PAHs and that their catalytic temperature corresponding to the maximum PAHs cracking rates is different. The PAHs cracking rate is up to 60.59% for Fe2O3 at 600°C and is 52.88% at 700°C for CaO. The catalytic temperature and particle size of the catalysts have a significant effect on PAHs cracking rate and CaO will lose the capability of decreasing 16 PAHs when the temperature is higher than 900°C. The possible cracking process of 16 PAHs is deduced by elaborately analyzing the cracking effect of the two catalysts on 16 different species of PAHs.
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One-pot preparation of glutathione-silica hybrid monolith for mixed-mode capillary liquid chromatography based on "thiol-ene" click chemistry.
J Chromatogr A
PUBLISHED: 03-21-2014
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A novel glutathione (GSH)-silica hybrid monolithic column synthesized via a combination of thiol-ene click reaction and one-pot process was described, where thiol-end GSH organic monomer and 2,2-azobisisobutyronitrile (AIBN) were mixed with hydrolyzed tetramethyloxysilane (TMOS) and ?-methacryloxypropyltrimethoxysilane (?-MAPS) and then introduced into a fused-silica capillary for simultaneous polycondensation and "thiol-ene" click reaction to form the GSH-silica hybrid monolith. The effects of the molar ratio of TMOS/?-MAPS, the amount of GSH, and the volume of porogen on the morphology, permeability and pore properties of the prepared GSH-silica hybrid monoliths were studied in detail. A uniform monolithic network with high porosity was obtained. A series of test compounds including alkylbenzenes, amides, and anilines were used to evaluate the retention behaviors of the GSH-silica hybrid monolithic column. The results demonstrated that the prepared GSH-silica hybrid monolith exhibited multiple interactions including hydrophobicity, hydrophilicity, as well as cation exchange interaction. The run-to-run, column-to-column and batch-to-batch reproducibilities of the GSH-silica hybrid monolith for phenols' retention were satisfactory with the relative standard deviations (RSDs) less than 1.3% (n=5), 2.6% (n=3) and 3.2% (n=3), respectively, indicating the effectiveness and practicability of the proposed method. In addition, the GSH-silica hybrid monolith was applied to the separation of nucleotides, peptides and protein tryptic digests, respectively. The successful applications suggested the potential of the GSH-silica hybrid monolith in complex sample analysis.
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Activity levels and expression of antioxidant enzymes in the ascorbate-glutathione cycle in artificially aged rice seed.
Plant Physiol. Biochem.
PUBLISHED: 03-06-2014
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Reactive oxygen species are the main contributors to seed deterioration. In order to study scavenging systems for reactive oxygen species in aged seed, we performed analyses using western blotting, real-time quantitative reverse-transcription polymerase chain reaction, high-performance liquid chromatography, and antioxidant enzyme activity analyses in artificially aged rice seeds (Oryza sativa L. cv. wanhua no.11). Aging seeds by storing them at 50 °C for 1, 9, or 17 months increased the superoxide radical and hydrogen peroxide levels and reduced the germination percentage from 99% to 92%, 55%, and 2%, respectively. The activity levels of superoxide dismutase (SOD), glutathione reductase (GR), and dehydroascorbate reductase (DHAR) did not change in aged seeds. In contrast, the activity levels of catalase (CAT), ascorbate peroxidase (APX), and monodehydroascorbate reductase (MDHAR) were significantly decreased in aged seeds, as were the expression of catalase and cytosolic ascorbate peroxidase protein. Transcript accumulation analysis showed that specific expression patterns were complex for each of the antioxidant enzyme types in the rice embryos. Overall, the expression of most genes was down-regulated, along with their protein expression. In addition, the reduction in the amount of ascorbate and glutathione was associated with the reduction in scavenging enzymes activity in aged rice embryos. Our data suggest that the depression of the antioxidant system, especially the reduction in the expression of CAT1, APX1 and MDHAR1, may be responsible for the accumulation of reactive oxygen species in artificially aged seed embryos, leading to a loss of seed vigor.
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Biomass fuels and coke plants are important sources of human exposure to polycyclic aromatic hydrocarbons, benzene and toluene.
Environ. Res.
PUBLISHED: 03-05-2014
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Large amounts of carcinogenic polycyclic aromatic hydrocarbons (PAHs), benzene and toluene (BT) might be emitted from incomplete combustion reactions in both coal tar factories and biomass fuels in rural China. The health effects arising from exposure to PAHs and BT are a concern for residents of rural areas close to coal tar plants. To assess the environmental risk and major exposure sources, 100 coke plant workers and 25 farmers in Qujing, China were recruited. The levels of 10 mono-hydroxylated PAHs (OH-PAHs), four BT metabolites and 8-hydroxy-2'-deoxyguanosine (8-OHdG) in the urine collected from the subjects were measured. The 8-OHdG levels in the urine were determined to evaluate the oxidative DNA damage induced by the PAHs and BT. The results showed that the levels of the OH-PAHs, particularly those of 1-hydroxynathalene and 1-hydroxypyrene, in the farmers were 1-7 times higher than those in the workers. The concentrations of the BT metabolites were comparable between the workers and farmers. Although the exact work location within a coke oven plant might affect the levels of the OH-PAHs, one-way ANOVA revealed no significant differences for either the OH-PAHs levels or the BT concentrations among the three groups working at different work sites. The geometric mean concentration (9.17µg/g creatinine) of 8-OHdG was significantly higher in the farmers than in the plant workers (6.27µg/g creatinine). The levels of 8-OHdG did not correlate with the total concentrations of OH-PAHs and the total levels of BT metabolites. Incompletely combusted biomass fuels might be the major exposure source, contributing more PAHs and BT to the local residents of Qujing. The estimated daily intakes (EDIs) of naphthalene and fluorene for all of the workers and most of the farmers were below the reference doses (RfDs) recommended by the U.S. Environmental Protection Agency (EPA), except for the pyrene levels in two farmers. However, the EDIs of benzene in the workers and local farmers ranged from 590 to 7239µg/day, and these levels were 2- to 30-fold higher than the RfDs recommended by the EPA. Biomass fuel combustion and industrial activities related to coal tar were the major sources of the PAH and BT exposure in the local residents. Using biomass fuels for household cooking and heating explains the higher exposure levels observed in the farmers relative to the workers at the nearby coal tar-related industrial facility.
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Comparison of Acoustic Radiation Force Impulse Imaging and Transient Elastography for Non-invasive Assessment of Liver Fibrosis in Patients with Chronic Hepatitis B.
Ultrasound Med Biol
PUBLISHED: 03-04-2014
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The aims of this study were to compare the performance of acoustic radiation force impulse (ARFI) elastography and transient elastography (TE) in the assessment of liver fibrosis in patients with chronic hepatitis B and to evaluate the impact of elevated alanine transaminase levels on liver stiffness assessment using ARFI elastography. One hundred eighty consecutive patients with chronic hepatitis B were enrolled in this study and evaluated with respect to histologic and biochemical features. All patients underwent ARFI elastography and TE. ARFI elastography and TE correlated significantly with histologically assessed fibrosis (r = 0.599, p < 0.001, for ARFI elastography; r = 0.628, p < 0.001, for TE) and necro-inflammatory activity (r = 0.591, p < 0.001, for ARFI elastography; r = 0.616, p < 0.001, for TE). Areas under the receiver operating characteristic curves for ARFI elastography and TE were 0.764 and 0.813 (p = 0.302, ?stage 2), 0.852 and 0.852 (p = 1.000, ?stage 3) and 0.825 and 0.799 (p = 0.655, S = 4), respectively. The optimum cutoff values for ARFI elastography were 1.63 m/s for stage ?2, 1.74 m/s for stage ?3 and 2.00 m/s for stage 4 in patients for whom alanine transaminase levels were evaluated. The cutoff values decreased to 1.24 m/s for ? stage 2, 1.32 m/s for ? stage 3 and 1.41 m/s for stage 4 in patients with normal alanine transaminase levels. ARFI elastography may be a reliable method for diagnosing the stage of liver fibrosis with diagnostic performance similar to that of TE in patients with chronic hepatitis B. In addition, liver stiffness values obtained with ARFI elastography, like those obtained with TE, may be influenced by alanine transaminase levels.
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Anthocyanin increases adiponectin secretion and protects against diabetes-related endothelial dysfunction.
Am. J. Physiol. Endocrinol. Metab.
PUBLISHED: 03-04-2014
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Adiponectin is an adipose tissue-secreted adipokine with beneficial effects on the cardiovascular system. In this study, we evaluated a potential role for adiponectin in the protective effects of anthocyanin on diabetes-related endothelial dysfunction. We treated db/db mice on a normal diet with anthocyanin cyanidin-3-O-?-glucoside (C3G; 2 g/kg diet) for 8 wk. Endothelium-dependent and -independent relaxations of the aorta were then evaluated. Adiponectin expression and secretion were also measured. C3G treatment restores endothelium-dependent relaxation of the aorta in db/db mice, whereas diabetic mice treated with an anti-adiponectin antibody do not respond. C3G treatment induces adiponectin expression and secretion in cultured 3T3 adipocytes through transcription factor forkhead box O1 (Foxo1). Silencing Foxo1 expression prevented C3G-stimulated induction of adiponectin expression. In contrast, overexpression of Foxo1-ADA promoted adiponectin expression in adipocytes. C3G activates Foxo1 by increasing its deacetylation via silent mating type information regulation 2 homolog 1 (Sirt1). Furthermore, purified anthocyanin supplementation significantly improved flow-mediated dilation (FMD) and increased serum adiponectin concentrations in patients with type 2 diabetes. Changes in adiponectin concentrations positively correlated with FMD in the anthocyanin group. Mechanistically, adiponectin activates cAMP-PKA-eNOS signaling pathways in human aortic endothelial cells, increasing endothelial nitric oxide bioavailability. These results demonstrate that adipocyte-derived adiponectin is required for anthocyanin C3G-mediated improvement of endothelial function in diabetes.
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Univariate and multivariate analyses for postoperative bleeding after nasal endoscopic surgery.
Acta Otolaryngol.
PUBLISHED: 03-03-2014
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Our study suggested that the major risk factors for postoperative bleeding after nasal endoscopic surgery (NES) included hypertension, long-term non-steroidal anti-inflammatory drugs (NSAIDs), and previous nasal surgery. The use of preoperative corticosteroids is a valuable measure for reducing postoperative bleeding after NES.
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SOX4 inhibits GBM cell growth and induces G0/G1 cell cycle arrest through Akt-p53 axis.
BMC Neurol
PUBLISHED: 02-26-2014
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BackgroundSOX4 is a transcription factor required for tissue development and differentiation in vertebrates. Overexpression of SOX4 has been reported in many cancers including glioblastoma multiforme (GBM), however, the underlying mechanism of actions has not been studied. In this study, we investigated the role of SOX4 in GBM.MethodsKaplan-Meier analysis was performed to assess the association between SOX4 expression levels and survival times in primary GBM samples. Cre/lox P system was used to generate gain or loss of SOX4 in GBM cells, and microarray analysis uncovered the regulation network of SOX4 in GBM cells.ResultsHigh SOX4 expression was significantly associated with good prognosis of primary GBMs. SOX4 inhibited the growth of GBM cell line LN229, A172G and U87MG, partly via the activation of p53-p21 signaling and down-regulation of phosphorylated AKT1. Gene expression profiling and subsequent gene ontology analysis showed that SOX4 influenced several key pathways including the Wnt/ beta-catenin and TGF-beta signaling pathways.ConclusionsOur study found that SOX4 acts as a tumor suppressor in GBM cells by induce cell cycle arrest and inhibiting cell growth.
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Overexpression of GmAKT2 potassium channel enhances resistance to soybean mosaic virus.
BMC Plant Biol.
PUBLISHED: 02-25-2014
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Soybean mosaic virus (SMV) is the most prevalent viral disease in many soybean production areas. Due to a large number of SMV resistant loci and alleles, SMV strains and the rapid evolution in avirulence/effector genes, traditional breeding for SMV resistance is complex. Genetic engineering is an effective alternative method for improving SMV resistance in soybean. Potassium (K+) is the most abundant inorganic solute in plant cells, and is involved in plant responses to abiotic and biotic stresses. Studies have shown that altering the level of K+ status can reduce the spread of the viral diseases. Thus K+ transporters are putative candidates to target for soybean virus resistance.
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Associations of retinol-binding protein 4 with oxidative stress, inflammatory markers, and metabolic syndrome in a middle-aged and elderly Chinese population.
Diabetol Metab Syndr
PUBLISHED: 02-18-2014
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Retinol-binding protein 4 (RBP4), a novel adipokine secreted by adipocytes and the liver, has elevated levels in type 2 diabetes mellitus (T2DM). However, its association with human metabolic diseases remains controversial. The present study was designed to investigate the associations of plasma RBP4 levels with oxidative stress, inflammatory markers, and metabolic syndrome (MetS) in a Chinese population.
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Polymer monolithic capillary column fabricated by using monodisperse iron oxide nanocrystal template to enhance the electrochromatographic separation of small molecules.
Electrophoresis
PUBLISHED: 02-17-2014
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Monodisperse iron oxide nanocrystals and organic solvents were utilized as coporogens in monolithic poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) capillary columns to afford stationary phases with enhanced electrochromatographic performance of small molecules. While the conventional monoliths using organic solvents only as a porogen exhibited poor resolution (Rs) <1.0 and low efficiency of 40?000-60?000 plates/m, addition of a small amount of nanocrystals to the polymerization mixture provided increased resolution (Rs > 3.0) and high efficiency ranged from 60?000 to 100?000 plates/m at the same linear velocity of 0.856 mm/s. It was considered that the mesopores introduced by the nanocrystals played an important role in the improvement of the monolith performance. This new strategy expanded the application range of the hydrophobic monoliths in the separation of polar alkaloids and narcotics. The successful applications demonstrated that the glycidyl methacrylate based monoliths prepared by using nanocrystal template are a good alternative for enhanced separation efficiency of small molecules.
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Fine-tuning of epigenetic regulation with respect to promoter CpG content in a cell type-specific manner.
Epigenetics
PUBLISHED: 02-12-2014
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Epigenetic regulation of gene expression is fundamental for cell type-specific gene expression. However, integrated comparative transcriptomic and epigenomic analyses in various adult primary differentiated cells remain underrepresented. We generated promoter landscapes of DNA methylation and three important histone methylation marks (H3K4me3, H3K9me2, and H3K27me3) in two primary cell types (B lymphocytes and liver) from adult mice. In line with previous studies, we also observed distinct H3K4me3 patterns at promoters dictated by CpG content in differentiated primary cells. We further explored the distribution of initiating RNA polymerase II and elongating RNA polymerase II across genes within different promoter classes, suggesting different rate-limiting steps at CpG-rich vs. CpG-poor genes. Examination of differentially expressed genes revealed that regulation of tissue-specific genes is closely related to gene function regardless of promoter type. Although repressive chromatin marks displayed differential preference to promoters based on CpG content, we observed fine-tuning of the pattern of association of these marks with specific promoter types in a cell type-specific manner. The distribution of H3K9me2 and H3K27me3, relative to CpG content, differed substantially between the two cell types. Cell-type specific accumulation of repressive chromatin marks was also observed at silent genes in both cell types, suggesting that differentiated primary cells may exhibit cell-type specificity in the distribution of repressive chromatin marks. Epigenetic regulation of gene expression and the association of specific histone marks with promoter sequence classes are fine-tuned in a cell type-specific manner. This unexpected finding underscores the value of extensive study of epigenetic marks across cell and tissue types.
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Obesity, rather than diet, drives epigenomic alterations in colonic epithelium resembling cancer progression.
Cell Metab.
PUBLISHED: 01-27-2014
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While obesity represents one of several risk factors for colorectal cancer in humans, the mechanistic underpinnings of this association remain unresolved. Environmental stimuli, including diet, can alter the epigenetic landscape of DNA cis-regulatory elements affecting gene expression and phenotype. Here, we explored the impact of diet and obesity on gene expression and the enhancer landscape in murine colonic epithelium. Obesity led to the accumulation of histone modifications associated with active enhancers at genomic loci downstream of signaling pathways integral to the initiation and progression of colon cancer. Meanwhile, colon-specific enhancers lost the same histone mark, poising cells for loss of differentiation. These alterations reflect a transcriptional program with many features shared with the program driving colon cancer progression. The interrogation of enhancer alterations by diet in colonic epithelium provides insights into the biology underlying high-fat diet and obesity as risk factors for colon cancer.
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Vitamin d-binding protein levels in plasma and gingival crevicular fluid of patients with generalized aggressive periodontitis.
Int J Endocrinol
PUBLISHED: 01-24-2014
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Vitamin D-binding protein (DBP) is the main transport protein of vitamin D and plays an important role in the immune system and host defenses. The purpose of this study was to measure DBP levels in plasma and gingival crevicular fluid (GCF) of patients with generalized aggressive periodontitis (GAgP), in comparison to healthy controls, with the goal of elucidating the relationship between DBP and GAgP. Fifty-nine GAgP patients and 58 healthy controls were recruited for the study; clinical parameters of probing depths (PD), bleeding index, and attachment loss (AL) were recorded. DBP levels were measured by enzyme-linked immunosorbent assay. From the results, GAgP patients had higher plasma DBP concentrations (P < 0.001) but lower GCF DBP concentrations (P < 0.001) than healthy controls. In GAgP group, after controlling the potential confounders of age, gender, smoking status, and BMI index, GCF DBP concentrations correlated negatively with PD (P < 0.001) and AL (P = 0.009). Within the limits of the study, we concluded that decreased GCF DBP level and increased plasma DBP level are associated with periodontitis.
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The methylation status of the platelet-derived growth factor-B gene promoter and its regulation of cellular proliferation following folate treatment in human glioma cells.
Brain Res.
PUBLISHED: 01-21-2014
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Platelet-derived growth factor-B (PDGF-B) is a growth factor that regulates cell migration, proliferation, and differentiation, and is involved in several physical and pathological processes. The overexpression of PDGF-B in glioma surgical samples revealed its effect on tumorigenesis. In this study, we determined that the expression of PDGF-B in 54 glioma samples varied among different grades and was correlated with the cell proliferation marker, Ki-67. Using pyrosequencing, we quantitatively assessed PDGF-B gene methylation levels and determined that hypomethylation promotes increased expression of PDGF-B in higher grade gliomas. Furthermore, we treated two glioma cell lines with a demethylating agent (5-aza-2'-deoxycitidine, 5-aza-dC) or a remethylating agent (folate) to alter the methylation status of PDGF-B. The epigenetic regulation of the PDGF-B gene not only modulated the expression levels of PDGF-B but also affected the cellular proliferation induced by TGF?-Smad activity and the PDGF-B peptide itself. Our work showed the importance of the methylation status of the PDGF-B gene promoter, and suggests that the epigenetic regulation of the PDGF-B gene may serve as a potential therapeutic target for the inhibition of glioma proliferation.
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Ag(I)-bovine serum albumin hydrosol-mediated formation of Ag3PO4/reduced graphene oxide composites for visible-light degradation of Rhodamine B solution.
J Colloid Interface Sci
PUBLISHED: 01-11-2014
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A cost-effective Ag(I)-bovine serum albumin (BSA) supramolecular hydrosol strategy was utilized to assemble Ag3PO4 nanospheres onto reduced graphene oxide (rGO) sheets. The obtained composites were characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy, UV-vis absorption spectroscopy and Fourier transform infrared spectroscopy. Compared with the pure Ag3PO4 crystals and Ag3PO4 particles prepared with Ag(I)-BSA hydrosol as precursor, the Ag3PO4/rGO composites obtained with different content of graphene oxide indicated improved visible-light-driven photocatalysis activity for the decomposition of Rhodamine B aqueous solution. The results pointed to the possibility of synthesizing graphene-based photocatalysts by metal ion-BSA hydrosol.
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Exposure to polycyclic aromatic hydrocarbons could cause their oxidative DNA damage: a case study for college students in Guangzhou, China.
Environ Sci Pollut Res Int
PUBLISHED: 01-09-2014
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Human exposure to carcinogenic polycyclic aromatic hydrocarbons (PAHs) in cigarette smoking might result in generation of reactive oxygen species (ROS) and induce formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG). This study was designed to examine whether levels of 8-OHdG are associated with levels of urinary metabolites of PAHs. Two groups (smokers and non-smokers) were recruited from college students in Guangzhou, China. Their urine samples were collected and analyzed for ten urinary mono-hydroxylated PAHs (OH-PAHs) and 8-OHdG by liquid chromatography equipped with tandem mass spectrometer (LC/MS/MS). Multiple linear regression analysis was performed to examine correlations between urinary levels of 8-OHdG and OH-PAHs. No significant difference was observed for creatinine-adjusted OH-PAHs between smokers and non-smokers. The levels of 8-OHdG between smokers and non-smokers were comparative. OH-PAH levels in this study were 2-50 times higher than those in populations from other countries and areas. The estimated daily intake (EDI; ?g/day) of PAHs ranged from 0.02 to 371.4, which were far lower than the reference doses (RfDs) specified by U.S. Environmental Protection Agency (EPA). Though smoking was a main factor, which affected the PAH exposure, it was not a dominant factor in the exposure to PAHs of Guangzhou college students. The environmental exposure could not be ignored. The sum concentrations of OH-PAHs (?OH-PAHs) had a dose-increase relationship with 8-OHdG both for smokers and non-smokers, especially for smokers. Though people in Guangzhou bore higher PAH hazards, the estimated environmental risk was still under safe ranges.
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A Simplified and Versatile System for the Simultaneous Expression of Multiple siRNAs in Mammalian Cells Using Gibson DNA Assembly.
PLoS ONE
PUBLISHED: 01-01-2014
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RNA interference (RNAi) denotes sequence-specific mRNA degradation induced by short interfering double-stranded RNA (siRNA) and has become a revolutionary tool for functional annotation of mammalian genes, as well as for development of novel therapeutics. The practical applications of RNAi are usually achieved by expressing short hairpin RNAs (shRNAs) or siRNAs in cells. However, a major technical challenge is to simultaneously express multiple siRNAs to silence one or more genes. We previously developed pSOS system, in which siRNA duplexes are made from oligo templates driven by opposing U6 and H1 promoters. While effective, it is not equipped to express multiple siRNAs in a single vector. Gibson DNA Assembly (GDA) is an in vitro recombination system that has the capacity to assemble multiple overlapping DNA molecules in a single isothermal step. Here, we developed a GDA-based pSOK assembly system for constructing single vectors that express multiple siRNA sites. The assembly fragments were generated by PCR amplifications from the U6-H1 template vector pB2B. GDA assembly specificity was conferred by the overlapping unique siRNA sequences of insert fragments. To prove the technical feasibility, we constructed pSOK vectors that contain four siRNA sites and three siRNA sites targeting human and mouse ?-catenin, respectively. The assembly reactions were efficient, and candidate clones were readily identified by PCR screening. Multiple ?-catenin siRNAs effectively silenced endogenous ?-catenin expression, inhibited Wnt3A-induced ?-catenin/Tcf4 reporter activity and expression of Wnt/?-catenin downstream genes. Silencing ?-catenin in mesenchymal stem cells inhibited Wnt3A-induced early osteogenic differentiation and significantly diminished synergistic osteogenic activity between BMP9 and Wnt3A in vitro and in vivo. These findings demonstrate that the GDA-based pSOK system has been proven simplistic, effective and versatile for simultaneous expression of multiple siRNAs. Thus, the reported pSOK system should be a valuable tool for gene function studies and development of novel therapeutics.
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Novel mutation in FBN1 causes ectopia lentis and varicose great saphenous vein in one Chinese autosomal dominant family.
Mol. Vis.
PUBLISHED: 01-01-2014
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To identify genetic defects in a Chinese family with ectopia lentis (EL) and varicose great saphenous vein (GSV) and to analyze the correlations between phenotype and genotype.
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Enhanced antitumor efficacy of an oncolytic herpes simplex virus expressing an endostatin-angiostatin fusion gene in human glioblastoma stem cell xenografts.
PLoS ONE
PUBLISHED: 01-01-2014
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Viruses have demonstrated strong potential for the therapeutic targeting of glioblastoma stem cells (GSCs). In this study, the use of a herpes simplex virus carrying endostatin-angiostatin (VAE) as a novel therapeutic targeting strategy for glioblastoma-derived cancer stem cells was investigated. We isolated six stable GSC-enriched cultures from 36 human glioblastoma specimens and selected one of the stable GSCs lines for establishing GSC-carrying orthotopic nude mouse models. The following results were obtained: (a) VAE rapidly proliferated in GSCs and expressed endo-angio in vitro and in vivo 48 h and 10 d after infection, respectively; (b) compared with the control gliomas treated with rHSV or Endostar, the subcutaneous gliomas derived from the GSCs showed a significant reduction in microvessel density after VAE treatment; (c) compared with the control, a significant improvement was observed in the length of the survival of mice with intracranial and subcutaneous gliomas treated with VAE; (d) MRI analysis showed that the tumor volumes of the intracranial gliomas generated by GSCs remarkably decreased after 10 d of VAE treatment compared with the controls. In conclusion, VAE demonstrated oncolytic therapeutic efficacy in animal models of human GSCs and expressed an endostatin-angiostatin fusion gene, which enhanced antitumor efficacy most likely by restricting tumor microvasculature development.
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QTLs for seed vigor-related traits identified in maize seeds germinated under artificial aging conditions.
PLoS ONE
PUBLISHED: 01-01-2014
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High seed vigor is important for agricultural production due to the associated potential for increased growth and productivity. However, a better understanding of the underlying molecular mechanisms is required because the genetic basis for seed vigor remains unknown. We used single-nucleotide polymorphism (SNP) markers to map quantitative trait loci (QTLs) for four seed vigor traits in two connected recombinant inbred line (RIL) maize populations under four treatment conditions during seed germination. Sixty-five QTLs distributed between the two populations were identified and a meta-analysis was used to integrate genetic maps. Sixty-one initially identified QTLs were integrated into 18 meta-QTLs (mQTLs). Initial QTLs with contribution to phenotypic variation values of R(2)>10% were integrated into mQTLs. Twenty-three candidate genes for association with seed vigor traits coincided with 13 mQTLs. The candidate genes had functions in the glycolytic pathway and in protein metabolism. QTLs with major effects (R(2)>10%) were identified under at least one treatment condition for mQTL2, mQTL3-2, and mQTL3-4. Candidate genes included a calcium-dependent protein kinase gene (302810918) involved in signal transduction that mapped in the mQTL3-2 interval associated with germination energy (GE) and germination percentage (GP), and an hsp20/alpha crystallin family protein gene (At5g51440) that mapped in the mQTL3-4 interval associated with GE and GP. Two initial QTLs with a major effect under at least two treatment conditions were identified for mQTL5-2. A cucumisin-like Ser protease gene (At5g67360) mapped in the mQTL5-2 interval associated with GP. The chromosome regions for mQTL2, mQTL3-2, mQTL3-4, and mQTL5-2 may be hot spots for QTLs related to seed vigor traits. The mQTLs and candidate genes identified in this study provide valuable information for the identification of additional quantitative trait genes.
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Upregulation of ?1-adrenoceptors is involved in the formation of gastric dysmotility in the 6-hydroxydopamine rat model of Parkinson's disease.
Transl Res
PUBLISHED: 01-01-2014
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Gastrointestinal dysmotility is one of the nonmotor symptoms of Parkinson's disease (PD). Gastroparesis and upregulated ?-adrenoceptors (?-ARs) have been reported in rats with bilateral microinjection of 6-hydroxydopamine (6-OHDA) in the substantia nigra, but the underlying mechanism is unclear. The aim of the current study is to investigate the role of ?-ARs in gastroparesis in 6-OHDA rats. Gastric motility was studied through strain gauge measurement. Immunofluorescence, real-time reverse transcription-polymerase chain reaction and Western blotting were performed to examine the expression of ?-ARs. Norepinephrine (NE) inhibited gastric motility in a dose-dependent fashion in both control and 6-OHDA rats, but much stronger adrenergic reactivity was observed in the 6-OHDA rats. The inhibition of gastric motility by NE in both control and 6-OHDA rats was not affected by tetrodotoxin, a neural sodium channel blocker. Blocking ?1-AR or ?2-AR did not affect the inhibition of strip contraction by NE in control rats, but ?1-AR blockage obviously enhanced the half maximal inhibitory concentration value of NE in 6-OHDA rats. Selective inhibition of ?3-AR blocked the effect of NE significantly in both control and 6-OHDA rats. The protein expression of ?1-AR, but not ?2-AR and ?3-AR in gastric muscularis externa was increased significantly in 6-OHDA rats. In conclusion, ?3-AR involves the regulation of gastric motility in control rats, whereas the upregulation of ?1-AR is responsible for enhanced NE reactivity in 6-OHDA rats and therefore is involved in the formation of gastroparesis. The effect of both ?1-AR and ?3-AR on gastric motility is independent of the enteric nervous system.
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Accuracy of Early DMSA Scan for VUR in Young Children With Febrile UTI.
Pediatrics
PUBLISHED: 12-23-2013
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To evaluate the accuracy of an acute (99m)Tc-dimercaptosuccinic acid (DMSA) scan in predicting dilating vesicoureteral reflux (VUR) among young children with a febrile urinary tract infection (UTI).
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Integrated microRNA network analyses identify a poor-prognosis subtype of gastric cancer characterized by the miR-200 family.
Clin. Cancer Res.
PUBLISHED: 12-18-2013
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Our aim was to investigate whether microRNAs can predict the clinical outcome of patients with gastric cancer (GC). We used integrated analysis of microRNA and mRNA expression profiles to identify GC microRNA subtypes and their underlying regulatory scenarios.
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Assessment and validation of density functional approximations for iron carbide and iron carbide cation.
J Phys Chem A
PUBLISHED: 12-18-2013
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Using quantum chemical approximations to understand and predict complex transition metal chemistry, such as catalytic processes and materials properties, is an important activity in modern computational chemistry. High-level theory can sometimes provide high-precision benchmarks for systems containing transition metals, and these benchmarks can be used to understand the reliability of less expensive quantum chemical approximations that are applicable to complex systems. Here, we studied the ionization potential energy of Fe and FeC and the bond dissociation energies of FeC and FeC(+) by 15 density functional approximations: M05, M06, M06-L, ?B97, ?B97X, ?B97X-D, ?-HCTHhyb, BLYP, B3LYP, M08-HX, M08-SO, SOGGA11, SOGGA11-X, M11, and M11-L. All of the functionals predict the correct spin state as the ground state of neutral iron atom, but five of them predict the wrong spin state for Fe(+). In the final analysis, four functionals, namely M11-L, ?-HCTHhyb, SOGGA11, and M06-L, have small mean unsigned errors when averaged over two bond dissociation energies and two ionization potentials. In fact, the results show that M11-L gives the smallest averaged mean unsigned error, i.e., M11-L is the most reliable density functional for these iron carbide systems among those studied.
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Serum leptin concentrations in Mongolian women.
Obes Res Clin Pract
PUBLISHED: 12-17-2013
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Summary: Objective: The aim of our study is to elucidate the association between leptin and obesity in Mongolian women.Method: Total 181 women participated in the study including 118 Mongolians and 63 Han. Body mass index (BMI) was calculated by weight (kg) divided by square height (m(2)). Percent body fat (%fat) was detected by bioelectrical impedance analysis (BIA). Fasting serum leptin was determined by ELISA.Result: The average BMI and %fat of Mongolian and Han women was 25.14 ± 4.48 kg/m(2), 24.30 ± 3.62 kg/m(2) and 36.10 ± 6.23%, 33.84 ± 5.98%, respectively. Fasting serum leptin level in obese women (BMI ? 25) was remarkably higher than in normal weight women (18.5 < BMI < 25) in Mongolian and Han ethnic groups (all P < 0.001). Fasting serum leptin level in Mongolian women had borderline significance compared with it in Han women (P = 0.049). Multiple linear regression models revealed that ethnicity, %fat and BMI were associated with serum leptin concentrations independent of age.Conclusion: In Mongolian and Han women, fasting serum leptin level was positively associated with BMI and %fat (all P < 0.001).
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Anthocyanin supplementation improves HDL-associated paraoxonase 1 activity and enhances cholesterol efflux capacity in subjects with hypercholesterolemia.
J. Clin. Endocrinol. Metab.
PUBLISHED: 11-27-2013
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Context and Objective:Paraoxonase 1 (PON1), an enzyme associated with high-density lipoprotein (HDL-PON1), is reported to have antioxidant and cardioprotective properties. The aim of the present study was to investigate the effects of anthocyanins on the HDL-PON1 activity and cholesterol efflux capacity in hypercholesterolemic subjects.Design and Participants:A total of 122 hypercholesterolemic subjects were given 160 mg of anthocyanins twice daily or placebo (n = 61 of each group) for 24 wk in a double-blind, randomized, placebo-controlled trial. Participants and investigators were masked to treatment allocation.Results:Anthocyanin consumption significantly increased HDL cholesterol and decreased LDL cholesterol concentrations compared with placebo (P < 0.018 and P<0.001, respectively). Anthocyanin supplementation also increased the activity of HDL-PON1 compared with placebo (P<0.001). Furthermore, cholesterol efflux capacity was increased more in the anthocyanin group (20.0% increase) than in the placebo group (0.2% increase) (P<0.001). The negative correlations established between HDL-PON1 activity and the levels of lipid hydroperoxides associated with HDL confirm the relationship between PON1 activity and lipid peroxidation of lipoproteins. Furthermore, a strong positive correlation was noted between increased HDL-PON1 activity and improved cholesterol efflux capacity both before and after adjustment for HDL cholesterol and apolipoprotein AI in anthocyanin-treated subjects (both P < 0.001). Inhibition of HDL-PON1 activity strongly prevented the antioxidant ability of HDL and attenuated the cholesterol efflux capacity of subjects from anthocyanin group.Conclusions:Our observations suggest that the alterations of PON1 activity by anthocyanin observed in hypercholesterolemic HDL reflect a shift to an improvement of cholesterol efflux capacity of HDL and may provide a link between anthocyanin and cardioprotective effects.
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MicroRNA-214 Antagonism Protects against Renal Fibrosis.
J. Am. Soc. Nephrol.
PUBLISHED: 10-24-2013
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Renal tubulointerstitial fibrosis is the common end point of progressive renal disease. MicroRNA (miR)-214 and miR-21 are upregulated in models of renal injury, but the function of miR-214 in this setting and the effect of its manipulation remain unknown. We assessed the effect of inhibiting miR-214 in an animal model of renal fibrosis. In mice, genetic deletion of miR-214 significantly attenuated interstitial fibrosis induced by unilateral ureteral obstruction (UUO). Treatment of wild-type mice with an anti-miR directed against miR-214 (anti-miR-214) before UUO resulted in similar antifibrotic effects, and in vivo biodistribution studies demonstrated that anti-miR-214 accumulated at the highest levels in the kidney. Notably, in vivo inhibition of canonical TGF-? signaling did not alter the regulation of endogenous miR-214 or miR-21. Whereas miR-21 antagonism blocked Smad 2/3 activation, miR-214 antagonism did not, suggesting that miR-214 induces antifibrotic effects independent of Smad 2/3. Furthermore, TGF-? blockade combined with miR-214 deletion afforded additional renal protection. These phenotypic effects of miR-214 depletion were mediated through broad regulation of the transcriptional response to injury, as evidenced by microarray analysis. In human kidney tissue, miR-214 was detected in cells of the glomerulus and tubules as well as in infiltrating immune cells in diseased tissue. These studies demonstrate that miR-214 functions to promote fibrosis in renal injury independent of TGF-? signaling in vivo and that antagonism of miR-214 may represent a novel antifibrotic treatment in the kidney.
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P-glycoprotein associates with Anxa2 and promotes invasion in multidrug resistant breast cancer cells.
Biochem. Pharmacol.
PUBLISHED: 09-17-2013
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Several recent studies have suggested that the acquisition of the multidrug resistance (MDR) phenotype is associated with elevated invasion and metastasis of tumor cells. P-glycoprotein (P-gp), the major determinant in the generation of the MDR phenotype, was reported to be correlated with a more aggressive phenotype and poor prognosis in many forms of malignancies. However, a clear understanding of the association is still lacking. We previously showed that Anxa2, a calcium-dependent phospholipid-binding protein, interacts with P-gp and contributes to the invasiveness of MDR breast cancer cells. In the present study, a strong positive correlation between MDR1 and Anxa2 mRNA expression in invasive breast cancer tissues during cancer progression was observed. In addition, exposure to adriamycin significantly enhanced motility in breast cancer cells and increased levels of P-gp and Anxa2. Moreover, inhibition of P-gp activity, using selective P-gp modulators, was found to significantly inhibit the invasive capacity of MCF-7/ADR cells without affecting the interaction and co-localization between P-gp and Anxa2. However, suppression of P-gp pump activity and knockdown of MDR1 expression both disrupted adriamycin-induced Anxa2 phosphorylation. Interestingly, P-gp was further demonstrated to interact with Src, a tyrosine kinase upstream of Anxa2. Taken together, our results indicate that P-gp may promote the invasion of MDR breast cancer cells by modulating the tyrosine phosphorylation of Anxa2. The interaction between Anxa2 and P-gp is possibly, at least in part, responsible for the association between MDR and invasive potential in breast cancer cells.
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De novo Mutations in the Cone-rod Homeobox Gene Associated with Leber Congenital Amaurosis in Chinese Patients.
Ophthalmic Genet.
PUBLISHED: 09-05-2013
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Abstract Background: The cone-rod homeobox (CRX) gene plays an important role in photoreceptor development. Recently, mutant alleles of the CRX gene have been associated with autosomal dominant Leber congenital amaurosis (LCA) and cone-rod dystrophy. The purpose of this study was to analyze the CRX mutations in a cohort of Chinese patients with LCA or early-onset severe retinal dystrophy (EOSRD) and to provide the clinical features of these patients. Methods: Patients with LCA or EOSRD were enrolled from 2003 to 2012. Detailed ocular examinations including optical coherence tomography (OCT) and standardized electrophysiology were performed. Genomic DNA was isolated with standard methods of genetic diagnosis. All three exons of CRX were amplified with PCR and screened for mutations through direct DNA sequencing. A total of 200 unrelated healthy Chinese subjects were screened to exclude nonpathogenic polymorphisms. Offspring-parent relationship was tested to confirm de novo mutation. Results: A total of 109 probands from 109 unrelated families were selected for mutation screening of the CRX gene. Two individuals with LCA were confirmed to carry de novo CRX mutations c.421delT (p.Ser141Pro fsX46) and c.571delT (p.Tyr191Met fsX3), respectively. The daughter of Case 1 also carried the same CRX mutation (c.421delT) and had LCA symptoms. Pigmentary retinopathy in the peripheral retina and macular atrophy were observed in the two probands. Macular atrophy without normal lamination structure was the retina phenotype under OCT. Conclusions: Two de novo mutations in CRX were found in Chinese patients with LCA. The CRX mutation might create a dominantly inherited trait.
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Extracellular matrix secretion by cardiac fibroblasts: role of microRNA-29b and microRNA-30c.
Circ. Res.
PUBLISHED: 09-04-2013
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Rationale: MicroRNAs (miRNAs), in particular miR-29b and miR-30c, have been implicated as important regulators of cardiac fibrosis. Objective: To perform a proteomics comparison of miRNA effects on extracellular matrix secretion by cardiac fibroblasts. Methods and Results: Mouse cardiac fibroblasts were transfected with pre-/anti-miR of miR-29b and miR-30c, and their conditioned medium was analyzed by mass spectrometry. miR-29b targeted a cadre of proteins involved in fibrosis, including multiple collagens, matrix metalloproteinases, and leukemia inhibitory factor, insulin-like growth factor 1, and pentraxin 3, 3 predicted targets of miR-29b. miR-29b also attenuated the cardiac fibroblast response to transforming growth factor-?. In contrast, miR-30c had little effect on extracellular matrix production but opposite effects regarding leukemia inhibitory factor and insulin-like growth factor 1. Both miRNAs indirectly affected cardiac myocytes. On transfection with pre-miR-29b, the conditioned medium of cardiac fibroblasts lost its ability to support adhesion of rat ventricular myocytes and led to a significant reduction of cardiac myocyte proteins (?-actinin, cardiac myosin-binding protein C, and cardiac troponin I). Similarly, cardiomyocytes derived from mouse embryonic stem cells atrophied under pre-miR-29 conditioned medium, whereas pre-miR-30c conditioned medium had a prohypertrophic effect. Levels of miR-29a, miR-29c, and miR-30c, but not miR-29b, were significantly reduced in a mouse model of pathological but not physiological hypertrophy. Treatment with antagomiRs to miR-29b induced excess fibrosis after aortic constriction without overt deterioration in cardiac function. Conclusions: Our proteomic analysis revealed novel molecular targets of miRNAs that are linked to a fibrogenic cardiac phenotype. Such comprehensive screening methods are essential to define the concerted actions of miRNAs in cardiovascular disease.
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Solution blowing of chitosan/PVA hydrogel nanofiber mats.
Carbohydr Polym
PUBLISHED: 08-29-2013
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Both nanofiber mats and hydrogel have their own advantages in wound healing. In this study, a novel hydrogel nanofiber mats were fabricated via solution blowing of chitosan and PVA solution, with various content of ethylene glycol diglycidyl ether (EGDE) as cross-linker. SEM observation showed that the fibers were several hundred nanometers in diameter with smooth surface and distributed randomly forming three-dimensional mats. The structure of the chitosan/PVA nanofibers was examined by FTIR and XPS, and the results showed that the cross-linking reaction occurred between EGDE and the hydroxyl groups. The mats could quickly hydrate in an aqueous environment to form hydrogel. Their value of equilibrate water absorption varied from 680 to 459% various content of EGDE. The nanofiber mats showed good bactericidal activity against Escherichia coli. The chitosan/PVA hydrogel nanofiber mats showed the combination advantages of nanofibrous mats and hydrogel dressing, and were suggested as potential application in wound healing.
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Protective Role of Statins in Patients With Acute Coronary Syndrome Aged >=75 Years With Low LDL-C Who Underwent Percutaneous Coronary Intervention.
Angiology
PUBLISHED: 08-28-2013
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The effect of statins in patients with acute coronary syndrome (ACS) at advanced age with lower low-density lipoprotein cholesterol (LDL-C) levels undergoing percutaneous coronary intervention (PCI) remains unknown. We evaluated the effect of statins in 220 Chinese patients with ACS aged ?75 years with low LDL-C undergoing PCI. Biomarkers were measured before and 6 hours after PCI, and patients were followed up for 1 year. Biomarkers in the statin group at 6 hours post-PCI were lower than controls (creatine kinase-myocardial band 14.2 ± 5.78 vs 47.3 ± 16.4 IU/L, P = .03; cardiac troponin I 0.36 ± 0.12 vs 1.33 ± 0.47 ng/mL, P = .01; and high-sensitivity C-reactive protein 7.6 ± 4.3 vs 13.6 ± 4.5 mg/L, P = .001, respectively). Significant differences were found in major adverse cardiac events at 1 year (P = .02-.01), while target lesion revascularization alone was less at 3 months between the 2 groups (P = .03). This study demonstrates that elderly patients with ACS having low LDL-C benefit from statins regardless of type, dosage, and duration of statin administration prior to PCI.
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Preparation of boronate-functionalized molecularly imprinted monolithic column with polydopamine coating for glycoprotein recognition and enrichment.
J Chromatogr A
PUBLISHED: 08-16-2013
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A novel imprinting strategy using reversible covalent complexation of glycoprotein was described for creating glycoprotein-specific recognition cavities on boronate-functionalized monolithic column. Based on it, a molecularly imprinted monolithic column was prepared by self-polymerization of dopamine (DA) on the surface of 4-vinylphenylboronic acid (VPBA)-based polymeric skeletons after reversible immobilization of horseradish peroxidase (HRP). Due to the combination of boronate affinity and surface imprinting of DA, the stable and accessible recognition sites in the as-prepared imprinted monolith could be obtained after the removal of the template, which facilitated the rebinding of the template and provided good reproducibility and lifetime of use. The recognition behaviors of proteins on the bare VPBA-based, HRP-imprinted and nonimprinted monolithic columns were evaluated in detail and the results showed that the HRP-imprinted monolith exhibited higher recognition ability toward the template than another two monolithic columns. Not only nonglycoproteins but also glycoproteins can be well separated with the HRP-imprinted monolith. In addition, the feasibility of the HRP-imprinted monolith, adopted as an in-tube solid phase microextraction (in-tube SPME), was further assessed by selective extraction and enrichment of HRP from human serum. The good results demonstrated its potential in glycoproteome analysis.
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Next generation sequencing-based molecular diagnosis of retinitis pigmentosa: identification of a novel genotype-phenotype correlation and clinical refinements.
Hum. Genet.
PUBLISHED: 06-07-2013
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Retinitis pigmentosa (RP) is a devastating form of retinal degeneration, with significant social and professional consequences. Molecular genetic information is invaluable for an accurate clinical diagnosis of RP due to its high genetic and clinical heterogeneity. Using a gene capture panel that covers 163 of the currently known retinal disease genes, including 48 RP genes, we performed a comprehensive molecular screening in a collection of 123 RP unsettled probands from a wide variety of ethnic backgrounds, including 113 unrelated simplex and 10 autosomal recessive RP (arRP) cases. As a result, 61 mutations were identified in 45 probands, including 38 novel pathogenic alleles. Interestingly, we observed that phenotype and genotype were not in full agreement in 21 probands. Among them, eight probands were clinically reassessed, resulting in refinement of clinical diagnoses for six of these patients. Finally, recessive mutations in CLN3 were identified in five retinal degeneration patients, including four RP probands and one cone-rod dystrophy patient, suggesting that CLN3 is a novel non-syndromic retinal disease gene. Collectively, our results underscore that, due to the high molecular and clinical heterogeneity of RP, comprehensive screening of all retinal disease genes is effective in identifying novel pathogenic mutations and provides an opportunity to discover new genotype-phenotype correlations. Information gained from this genetic screening will directly aid in patient diagnosis, prognosis, and treatment, as well as allowing appropriate family planning and counseling.
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Differential expression of Nad(P)H oxidase isoforms and the effects of atorvastatin on cardiac remodeling in two-kidney two-clip hypertensive rats.
Pharmazie
PUBLISHED: 05-25-2013
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The NADPH oxidases (Noxes) are a family of ROS (reactive oxygen species)-generating enzymes which play a critical role in the development of cardiac remodeling associated with heart failure. The Noxes of their catalytic isoforms include multiple homologues in cardiovascular cells with wide range tissue distribution. It is still unclear which Noxes represent the major enzymatic source of ROS in the heart and play a predominant role in cardiac hypertrophy. In this study we investigated the differential expression changes of NAD(P)H oxidase P47phox isoform and Nox homologues in left ventricle and the effects of atorvastatin on cardiac remodeling in two-kidney two-clip(2K2C) hypertensive rats. The mRNA and protein expression of Nox2, Nox4 and P47phox showed a sustained increase at 4, 8, 12 weeks after surgery in 2K2C rats. Administration of atorvastatin attenuated cardiac dysfunction, hypertrophy and fibrosis of 2K2C rats. However, atorvastatin treatment had no effects on BP regulation. Further studies revealed that atorvastatin inhibited the increased expression of Nox2, Nox4, P47phox as well as 02"- production in 2K2C hypertensive rats. These findings indicate that Nox2, Nox4 and P47phox play a crucial role in the development of cardiac remodeling in the 2K2C hypertensive rats. Atorvastatin, independent of BP control, exerts anti-remodeling effects partially by inhibition of NAD(P)H oxidase-mediated cardiac oxidative stress.
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Jatrorrhizine hydrochloride inhibits the proliferation and neovascularization of C8161 metastatic melanoma cells.
Anticancer Drugs
PUBLISHED: 05-23-2013
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Malignant melanoma is the most aggressive form of skin cancer. Although various antimelanoma approaches have been used in the clinics to treat the disease over the last three decades, none of the drugs significantly prolonged the survival of metastatic melanoma patients; hence, effective drugs against metastatic melanoma are highly desired. In this study, we explored an antimetastatic melanoma agent derived from traditional Chinese medicinal herbs and found that jatrorrhizine hydrochloride (JH), an active component of the traditional Chinese medicinal herb Coptis chinensis, inhibited the proliferation and neovascularization of C8161 human metastatic melanoma cells. JH suppressed C8161 cell proliferation in a dose-dependent manner, with a half-maximal inhibitory concentration of 47.4±1.6 ?mol/l; however, it did not induce significant cellular apoptosis at doses up to 320 ?mol/l. Mechanistic studies showed that JH-induced C8161 cell cycle arrest at the G0/G1 transition, which was accompanied by overexpression of the cell cycle-suppressive genes p21 and p27 at higher doses. Moreover, JH reduced C8161 cell-mediated neovascularization in vitro and in vivo and impeded the expression of the gene for VE-cadherin, a key protein in tumor vasculogenic mimicry and angiogenesis. Taken together, the effective inhibitory effects of JH on metastatic melanoma cell proliferation and neovascularization with low toxicity suggest that JH is a potential new antimelanoma drug candidate.
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Next-generation sequencing-based molecular diagnosis of a Chinese patient cohort with autosomal recessive retinitis pigmentosa.
Invest. Ophthalmol. Vis. Sci.
PUBLISHED: 05-11-2013
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Retinitis pigmentosa (RP) is a highly heterogeneous genetic disease; therefore, an accurate molecular diagnosis is essential for appropriate disease treatment and family planning. The prevalence of RP in China had been reported at 1 in 3800, resulting in an estimated total of 340,000 Chinese RP patients. However, genetic studies of Chinese RP patients have been very limited. To date, no comprehensive molecular diagnosis has been done for Chinese RP patients. With the emergence of next-generation sequencing (NGS), comprehensive molecular diagnosis of RP is now within reach. The purpose of this study was to perform the first NGS-based comprehensive molecular diagnosis for Chinese RP patients.
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Detection of Eight Periodontal Microorganisms and Distribution of Porphyromonas gingivalis fimA Genotypes in Chinese Patients With Aggressive Periodontitis.
J. Periodontol.
PUBLISHED: 05-07-2013
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Background: The microbiologic feature of aggressive periodontitis (AgP) in Chinese patients has not yet been determined. This study aims to investigate the prevalence of eight periodontal microorganisms and the distribution of the Porphyromonas gingivalis fimA genotype in a cohort of Chinese patients with AgP. Methods: Saliva and pooled subgingival plaque samples were collected from 81 patients with AgP (25 with incisor-first molar type and 56 with generalized type [GAgP]) and 34 periodontally healthy controls. Eight periodontal microorganisms, including Aggregatibacter actinomycetemcomitans, P. gingivalis, Tannerella forsythia, Treponema denticola, Campylobacter rectus, Prevotella intermedia, Prevotella nigrescens, and Fusobacterium nucleatum were detected in these samples by the polymerase chain reaction (PCR). In addition, the distribution of fimA genotypes was assessed in P. gingivalis-positive individuals by PCR. Results: The prevalence of P. gingivalis, T. forsythia, T. denticola, C. rectus, P. intermedia, F. nucleatum, and A. actinomycetemcomitans in patients with AgP was significantly higher than that in healthy controls. The prevalence of A. actinomycetemcomitans in patients with GAgP was relatively low (30.4%) compared with other pathogens. Results of logistic regression analysis showed that younger patients were more likely to harbor A. actinomycetemcomitans (odds ratio = 2.85). Type II was the most prevalent fimA genotype of P. gingivalis in patients with AgP. Conclusions: P. gingivalis, T. forsythia, T. denticola, C. rectus, P. intermedia, and F. nucleatum were the predominant periodontal pathogens of patients with GAgP in China. Type II of fimA was the most prevalent genotype of P. gingivalis in patients with AgP. The prevalence of A. actinomycetemcomitans in patients with GAgP was relatively low.
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Aberrant upregulation of LRRC1 contributes to human hepatocellular carcinoma.
Mol. Biol. Rep.
PUBLISHED: 04-29-2013
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Loss of apico-basal polarity often results in a malignant phenotype in epithelial tissues. Aberrant expression of polarity mediator proteins is closely associated with this process. LRRC1/LANO, a putative cell polarity regulator, was previously screened from our gene expression profiling in which its expression was significantly upregulated in hepatocellular carcinoma (HCC). In the present study, we provide evidences that LRRC1 plays a potential oncogenic function in HCC. Consistent with the microarray data, quantitative real-time PCR results showed LRRC1 was aberrantly upregulated in 37/56 (66.1 %, more than twofolds) of HCC specimens compared with adjacent non-cancerous livers. Furthermore, the cellular expression of LRRC1 in all HCC cell lines examined exhibited much higher level than that in normal adult liver tissue. Functional analyses revealed that overexpression of LRRC1 promoted, while knockdown of LRRC1 by RNA interference inhibited the growth and colony formation of HCC cells. Importantly, enhanced expression of LRRC1 conferred NIH3T3 cells the ability of cell transformation. In a xenograft tumor model, we found LRRC1 overexpression increased the tumorigenicity of HCC cells. Thus, our collective findings suggest that LRRC1 contributes to HCC development, and may be a potential target for therapeutic intervention in this disease.
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Mutation analysis of the SRD5A2, AR and SF-1 genes in 52 Chinese boys with hypospadias.
J. Pediatr. Endocrinol. Metab.
PUBLISHED: 04-18-2013
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Abstract Objectives: To investigate the prevalence of genetic mutations in steroid 5?-reductase-2 (SRD5A2), androgen receptor (AR) and steroidogenic factor-1 (SF-1) in Chinese children with hypospadias, and to also explore the possible underlying molecular mechanisms of this disease. Methods: A total of 52 boys with hypospadias were enrolled. Mutational analyses of the SRD5A2, AR and SF-1 genes were performed by direct sequencing. Results: SRD5A2 gene mutations were found in 13.5% (7/52 cases), including five compound heterozygotic and two homozygotic mutations. One novel heterozygotic SF-1 gene mutation was identified in a patient with perineal hypospadias and cryptorchidism, the patients mother also had the same mutation. No mutation was found in the AR gene. The clinical manifestations of patients with mutations in SRD5A2 or SF-1 varied. Conclusions: In Chinese patients, SRD5A2 gene mutations were, relatively, frequently associated with hypospadias. The SF-1 gene may be another candidate gene for hypospadias. In contrast, AR gene mutations are not commonly associated with this condition.
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Deregulated microRNAs in CD4(+) T cells from individuals with latent tuberculosis versus active tuberculosis.
J. Cell. Mol. Med.
PUBLISHED: 04-14-2013
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The mechanisms of latent tuberculosis (TB) infection remain elusive. Roles of microRNA (miRNA) have been highlighted in pathogen-host interactions recently. To identify miRNAs involved in the immune response to TB, expression profiles of miRNAs in CD4(+) T cells from patients with latent TB, active TB and healthy controls were investigated by microarray assay and validated by RT-qPCR. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to analyse the significant functions and involvement in signalling pathways of the differentially expressed miRNAs. To identify potential target genes for miR-29, interferon-? (IFN-?) mRNA expression was measured by RT-qPCR. Our results showed that 27 miRNAs were deregulated among the three groups. RT-qPCR results were generally consistent with the microarray data. We observed an inverse correlation between miR-29 level and IFN-? mRNA expression in CD4(+) T cells. GO and KEGG pathway analysis showed that the possible target genes of deregulated miRNAs were significantly enriched in mitogen-activated protein kinase signalling pathway, focal adhesion and extracellular matrix receptor interaction, which might be involved in the transition from latent to active TB. In all, for the first time, our study revealed that some miRNAs in CD4(+) T cells were altered in latent and active TB. Function and pathway analysis highlighted the possible involvement of miRNA-deregulated mRNAs in TB. The study might help to improve understanding of the relationship between miRNAs in CD4(+) T cells and TB, and laid an important foundation for further identification of the underlying mechanisms of latent TB infection and its reactivation.
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Collision tumors of the sella: coexistence of pituitary adenoma and craniopharyngioma in the sellar region.
World J Surg Oncol
PUBLISHED: 04-11-2013
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Collision tumors of the sellar region are relatively uncommon and consist mainly of more than one type of pituitary adenoma or a cyst or cystic tumor. The association of a pituitary adenoma and a craniopharyngioma is particularly rare. This study describes a rare occurrence in which a pituitary adenoma and a craniopharyngioma coexisted in the sellar region. The case involves a 47-year-old woman who underwent transsphenoidal surgery with subtotal tumor resection and reoperation using an interhemispheric transcallosal approach for total microsurgical resection of the tumor because the visual acuity in her left eye had re-deteriorated. Histopathological and immunohistochemical examinations of the excised tissue revealed a pituitary adenoma in the first operation and a craniopharyngioma in the second operation. Retrospective analysis found the coexistence of a pituitary adenoma and a craniopharyngioma, known as a collision tumor. Instead of the transsphenoidal approach, a craniotomy should be performed, to explore the suprasellar region.
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