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Find video protocols related to scientific articles indexed in Pubmed.
Discovery of systematic responses and potential biomarkers induced by ochratoxin A using metabolomics.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess
PUBLISHED: 09-25-2014
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Ochratoxin A (OTA) is known to be nephrotoxic and hepatotoxic in rodents when exposed orally. To understand the systematic responses to OTA exposure, GC-MS- and (1)H-NMR-based metabolomic techniques together with histopathological assessments were applied to analyse the urine and plasma of OTA-exposed rats. It was found that OTA exposure caused significant elevation of amino acids (alanine, glycine, leucine etc.), pentose (ribose, glucitol, xylitol etc.) and nucleic acid metabolites (pseudouridine, adenosine, uridine). Moreover, myo-inositol, trimethylamine-oxide (TMAO), pseudouridine and leucine were identified as potential biomarkers for OTA toxicity. The primary pathways included the pentose phosphate pathway (PPP), the Krebs cycle (TCA), the creatine pathway and gluconeogenesis. The activated PPP was attributed to the high requirements for nicotinamide adenine dinucleotide phosphate (NADPH), which is involved in OTA metabolism through cytochrome P450. The elevated gluconeogenesis and TCA suggest that energy metabolism was involved. The up-regulated synthesis of creatinine reveals the elevated catabolism of proteins. These findings provide an overview of systematic responses to OTA exposure and metabolomic insight into the toxicological mechanism of OTA.
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Bioactive metabolites produced by the endophytic fungus Phomopsis sp. YM355364.
Nat Prod Commun
PUBLISHED: 07-17-2014
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A new compound, 16-acetoxycytosporone B (1), along with four known ones, dankasterone A (2), dankasterone B (3), 3beta,5alpha,9alpha-trihydroxy-(22E,24R)-ergosta-7,22-dien-6-one (4), and cyclonerodiol oxide (5), were isolated from Phomopsis sp. YM355364, an endophytic fungus of Aconitum carmichaeli. Their structures were characterized by spectral analysis. Compound 2 exhibited significant inhibitory activity against influenza A/Thailand/Kan353/2004(H5N1) pseudovirus with all IC50 value of 3.56 microM. Compounds 1, 2, and 4 showed either moderate or weak antifungal activities against four pathogenic fungi.
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Synthesis of gold nanorods and their functionalization with bovine serum albumin for optical hyperthermia.
J Biomed Nanotechnol
PUBLISHED: 07-15-2014
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Although gold nanorods (GNRs) have been investigated extensively for optical hyperthermia therapies, the synthesis of rods is far from ideal. In this report, we optimized the synthesis of gold nanorods using hydroquinone as a reducing agent. Compared with the GNRs prepared by traditional ways, the as-synthesized rods have a flexibly tunable size and wider range of longitudinal surface plasmon resonance (LSPR). Furthermore, a series of small-length gold nanorods with length ranging from 30 to 90 nm were synthesized and they are more suitable for in vivo biomedical applications. Finally, we exploited a convenient approach for preparing water-soluble GNRs with less toxicity, better dispersion and flexible functionalization by exchanging hexadecyltrimethylammonium bromide (CTAB) on the surface of the rods with carboxylated bovine serum albumin (BSA) derivative, the BSA modified GNRs showed significant anticancer efficacy through near infrared (NIR) hyperthermia. We believe that the as-prepared gold nanorods will find promising applications in biomedical fields, especially in cancer therapy.
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Hydrogen-rich water reestablishes ros homeostasis but exerts differential effects on anthocyanin synthesis in two varieties of radish sprouts under UV-A irradiation.
J. Agric. Food Chem.
PUBLISHED: 06-30-2014
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The aims of the study were to investigate whether hydrogen gas (H2) was involved in regulation of anthocyanin biosynthesis in two contrasting radish (Raphanus sativus L.) varieties (low [LA] and high [HA] level of anthocyanin) under UV irradiation. The results showed that hydrogen-rich water (HRW) significantly blocked the UV-A-induced increase of H2O2 and O2(•-) accumulation, and enhanced the UV-A-induced increase of superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities in LA and HA. Furthermore, UV-A-induced increase of anthocyanin and total phenols was further enhanced only in HA sprouts cotreated with HRW. LC-MS/MS analysis showed that five anthocyanidins existed in HA sprouts, but only two in LA sprouts. Meanwhile, the cyanidin was the most abundant anthocyanidin in HA, and the cyanidin was 2-fold higher cotreated with HRW than UV-A. Molecular analyses showed that the anthocyanin biosynthesis-related genes were upregulated significantly in both HA (in particular) and LA sprouts treated with HRW plus UV-A. These data imply that HRW reestablishes reactive oxygen species homeostasis in both LA and HA, but exerts different effects on anthocyanin accumulation between them under UV-A.
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Combination of metagenomics and culture-based methods to study the interaction between ochratoxin a and gut microbiota.
Toxicol. Sci.
PUBLISHED: 06-27-2014
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Gut microbiota represent an important bridge between environmental substances and host metabolism. Here we reported a comprehensive study of gut microbiota interaction with ochratoxin A (OTA), a major food-contaminating mycotoxin, using the combination of metagenomics and culture-based methods. Rats were given OTA (0, 70, or 210 ?g/kg body weight) by gavage and fecal samples were collected at day 0 and day 28. Bacterial genomic DNA was extracted from the fecal samples and both 16S rRNA and shotgun sequencing (two main methods of metagenomics) were performed. The results indicated OTA treatment decreased the within-subject diversity of the gut microbiota, and the relative abundance of Lactobacillus increased considerably. Changes in functional genes of gut microbiota including signal transduction, carbohydrate transport, transposase, amino acid transport system, and mismatch repair were observed. To further understand the biological sense of increased Lactobacillus, Lactobacillus selective medium was used to isolate Lactobacillus species from fecal samples, and a strain with 99.8% 16S rRNA similarity with Lactobacillus plantarum strain PFK2 was obtained. Thin-layer chromatography showed that this strain could absorb but not degrade OTA, which was in agreement with the result in metagenomics that no genes related to OTA degradation increased. In conclusion, combination of metagenomics and culture-based methods can be a new strategy to study intestinal toxicity of toxins and find applicable bacterial strains for detoxification. When it comes to OTA, this kind of mycotoxin can cause compositional and functional changes of gut microbiota, and Lactobacillus are key genus to detoxify OTA in vivo.
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Two new butenolides produced by an Actinomycete Streptomyces sp.
Chem. Biodivers.
PUBLISHED: 06-18-2014
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Two new butenolides, (4S)-4,10-dihydroxydodec-2-en-1,4-olide (1) and (4S)-4,8,10-trihydroxy-10-methyldodec-2-en-1,4-olide (2), together with three known compounds, MKN-003B (3), MKN-003C (4), and cyclo(Ala-Leu) (5), were isolated from the culture broth of a bacterium of the genus Streptomyces derived from soil environment. The structures of these compounds were elucidated on the basis of spectroscopic analysis. The inhibitory activities of the butenolides against eight pathogenic fungi were evaluated. All of the butenolides showed moderate-or-weak antifungal activities in a broth microdilution assay.
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Diffusion tensor imaging of the cerebellum in patients after heat stroke.
Acta Neurol Belg
PUBLISHED: 06-13-2014
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To explore the application value of cerebellar diffusion tensor imaging (DTI) in patients after heat stroke (HS). Eleven patients after HS with a score of 3-9 in Glasgow Coma Scale (GCS) and seven age-matched healthy volunteers were selected to undergo MR examinations during the same hot summer. The MR studies including DTI were performed with a 1.5 T scanner. Fractional anisotropy (FA) values of normal-appearing cerebellar white and gray matter were measured and the differences between the two groups were evaluated with Mann-Whitney U test. The FA value of normal-appearing cerebellar white matter in patients after HS was found to be decreased compared to normal control subjects (652.5 ± 86.1 vs 769.5 ± 58.4, p = 0.025). The FA value of normal-appearing cerebellar gray matter in patients after HS was found to be decreased compared to normal control subjects (158.8 ± 27.9 vs 187.5 ± 15.8, p = 0.040). Neural damage of the cerebellum induced by HS may be effectively evaluated by DTI with the decrease of FA value in normal-appearing cerebellum structures.
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Preparation of oxidized agar and characterization of its properties.
Carbohydr Polym
PUBLISHED: 06-10-2014
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A series of oxidized agars with different carboxyl content were prepared, and their properties were determined and analyzed. The results showed that the gelling temperature, the optical rotation and the apparent viscosity of the agar solution, and the melting temperature, the strength, the hardness, the fracturability, the springiness, the chewiness and the gumminess of agar gel all decreased except that the cohesiveness increased after oxidation. The gel skeleton structures of agar before and after oxidation were all of the porous network structures, but the pores of gel skeleton structure became smaller and denser after oxidation.
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Antiviral C-25 epimers of 26-acetoxy steroids from the South China Sea gorgonian Echinogorgia rebekka.
J. Nat. Prod.
PUBLISHED: 06-02-2014
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Four new steroids with an acetoxy linked at the end of the side chain, echrebsteroids A-D (1-4) were obtained from the South China Sea gorgonian Echinogorgia rebekka. The absolute configurations of 1-3 were determined using the modified Mosher's method on the hydrolysis products. The isolation of the pair of epimers (2 and 3) represents the first reported separation of C-25 epimers of 26-acetoxy steroids. The (25R)-epimer (3) exhibited promising antiviral activity against respiratory syncytial virus with an IC50 value of 0.19 ?M and a comparatively higher therapeutic ratio (TC50/IC50 = 128).
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Synthesis of LiYF4:Yb, Er upconversion nanoparticles and its fluorescence properties.
J Nanosci Nanotechnol
PUBLISHED: 04-18-2014
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LiYbF4:Yb, Er nanoparticles have been successfully synthesized by thermal decomposition of multiple trifluoroacetic acid salts. The SEM and TEM results show the size of the LiYF4:Yb, Er nanoparticles is about 100 nm in diagonal line, and the morphology of the LiYF4:Yb, Er nanoparticles is highly uniform with octahedral structure. Under the excitation of 980 nm, the LiYF4:Yb, Er nanoparticles have higher upconversion luminescence efficiency compared with that of NaYF4:Yb, Er. The results indicate that the as-prepared LiYbF4:Yb, Er nanoparticles may have potential applications in bio-probes and displays.
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The preparation and application of N-terminal 57 amino acid protein of the follicle-stimulating hormone receptor as a candidate male contraceptive vaccine.
Asian J. Androl.
PUBLISHED: 04-10-2014
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Follicle-stimulating hormone receptor (FSHR), which is expressed only on Sertoli cells and plays a key role in spermatogenesis, has been paid attention for its potential in male contraception vaccine research and development. This study introduces a method for the preparation and purification of human FSHR 57-amino acid protein (FSHR-57aa) as well as determination of its immunogenicity and antifertility effect. A recombinant pET-28a(+)-FSHR-57aa plasmid was constructed and expressed in Escherichia coli strain BL21 Star TM (DE3) and the FSHR-57aa protein was separated and collected by cutting the gel and recovering activity by efficient refolding dialysis. The protein was identified by Western blot and high-performance liquid chromatography analysis with a band of nearly 7 kDa and a purity of 97.4%. Male monkeys were immunized with rhFSHR-57aa protein and a gradual rising of specific serum IgG antibody was found which reached a plateau on day 112 (16 weeks) after the first immunization. After mating of one male with three female monkeys, the pregnancy rate of those mated with males immunized against FSHR-57aa was significantly decreased while the serum hormone levels of testosterone and estradiol were not disturbed in the control or the FSHR-57aa groups. By evaluating pathological changes in testicular histology, we found that the blood-testis barrier remained intact, in spite of some small damage to Sertoli cells. In conclusion, our study demonstrates that the rhFSHR-57aa protein might be a feasible male contraceptive which could affect sperm production without disturbing hormone levels.
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Cyclonerol derivatives from Trichoderma longibrachiatum YM311505.
Nat Prod Commun
PUBLISHED: 04-03-2014
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A new sesquiterpene, 10,11-dihydrocyclonerotriol (1), together with two known compounds, catenioblin C (2) and sohirnone A (3), were isolated from Trichoderma longibrachiatum YM311505, an endophytic fungus of Azadirachta indica. Their structures were characterized and identified by spectral analysis. Compounds 1-3 exhibited antifungal activities against Pyricularia oryzae and Candida albicans.
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MicroRNA profiling of rats with ochratoxin A nephrotoxicity.
BMC Genomics
PUBLISHED: 03-14-2014
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Nephrotoxicity is the most prominent one among the various toxicities of ochratoxin A (OTA). MicroRNAs (miRNAs) are small non-coding RNAs that have an impact on a wide range of biological processes by regulating gene expression at post-transcriptional level or protein systhesis level. The objective of this study is to analyze miRNA profiling in the kidneys of rats gavaged with OTA.
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Heritability and genomics of gene expression in peripheral blood.
Nat. Genet.
PUBLISHED: 03-14-2014
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We assessed gene expression profiles in 2,752 twins, using a classic twin design to quantify expression heritability and quantitative trait loci (eQTLs) in peripheral blood. The most highly heritable genes (?777) were grouped into distinct expression clusters, enriched in gene-poor regions, associated with specific gene function or ontology classes, and strongly associated with disease designation. The design enabled a comparison of twin-based heritability to estimates based on dizygotic identity-by-descent sharing and distant genetic relatedness. Consideration of sampling variation suggests that previous heritability estimates have been upwardly biased. Genotyping of 2,494 twins enabled powerful identification of eQTLs, which we further examined in a replication set of 1,895 unrelated subjects. A large number of non-redundant local eQTLs (6,756) met replication criteria, whereas a relatively small number of distant eQTLs (165) met quality control and replication standards. Our results provide a new resource toward understanding the genetic control of transcription.
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Molecular characterisation of a sodium channel gene and identification of a Phe1538 to Ile mutation in citrus red mite, Panonychus citri.
Pest Manag. Sci.
PUBLISHED: 03-02-2014
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The citrus red mite, Panonychus citri (McGregor), is regarded as one of the most serious citrus pests in many countries and has developed high resistance to pyrethroids as a result of the intensive use of these acaricides.
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Prostaglandin E? receptor EP2 mediates Snail expression in hepatocellular carcinoma cells.
Oncol. Rep.
PUBLISHED: 02-12-2014
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Prostaglandin E2 (PGE2) has been shown to influence cell invasion and metastasis in several types of cancer, including hepatocellular carcinoma (HCC). however, the molecular mechanisms underlying it remain to be further elucidated. Snail, as one of key inducers of epithelial-mesenchymal transition (EMT), plays pivotal roles in HCC invasion and metastasis. The present study was designed to evaluate the possible signaling pathways through which PGE2 regulates Snail protein expression in HCC cell lines. PGE2 markedly enhanced Huh-7 cell invasion and migration ability by upregulating the expression level of Snail protein, and EP2 receptor played an important role in this process. Src, EGFR, Akt and mTOR were all activated and involved in the regulation of snail protein expression. Our findings suggest that PGE2 could upregulate the expression level of Snail protein through the EP2/Src/EGFR/Akt/mTOR pathway in Huh-7 cells, which promotes HCC cell invasion and migration.
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Exposure to diflubenzuron results in an up-regulation of a chitin synthase 1 gene in citrus red mite, Panonychus citri (Acari: Tetranychidae).
Int J Mol Sci
PUBLISHED: 01-27-2014
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Chitin synthase synthesizes chitin, which is critical for the arthropod exoskeleton. In this study, we cloned the cDNA sequences of a chitin synthase 1 gene, PcCHS1, in the citrus red mite, Panonychus citri (McGregor), which is one of the most economically important pests of citrus worldwide. The full-length cDNA of PcCHS1 contains an open reading frame of 4605 bp of nucleotides, which encodes a protein of 1535 amino acid residues with a predicted molecular mass of 175.0 kDa. A phylogenetic analysis showed that PcCHS1 was most closely related to CHS1 from Tetranychus urticae. During P. citri development, PcCHS1 was constantly expressed in all stages but highly expressed in the egg stage (114.8-fold higher than in the adult). When larvae were exposed to diflubenzuron (DFB) for 6 h, the mite had a significantly high mortality rate, and the mRNA expression levels of PcCHS1 were significantly enhanced. These results indicate a promising use of DFB to control P. citri, by possibly acting as an inhibitor in chitin synthesis as indicated by the up-regulation of PcCHS1 after exposure to DFB.
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Environmental and genetic contributors to salivary testosterone levels in infants.
Front Endocrinol (Lausanne)
PUBLISHED: 01-01-2014
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Transient activation of the hypothalamic-pituitary-gonadal axis in early infancy plays an important role in male genital development and sexual differentiation of the brain, but factors contributing to individual variation in testosterone levels during this period are poorly understood. We measured salivary testosterone levels in 222 infants (119 males, 103 females, 108 singletons, 114 twins) between 2.70 and 4.80?months of age. We tested 16 major demographic and medical history variables for effects on inter-individual variation in salivary testosterone. Using the subset of twins, we estimated genetic and environmental contributions to salivary testosterone levels. Finally, we tested single nucleotide polymorphisms (SNPs) within ±5?kb of genes involved in testosterone synthesis, transport, signaling, and metabolism for associations with salivary testosterone using univariate tests and random forest (RF) analysis. We report an association between 5?min APGAR scores and salivary testosterone levels in males. Twin modeling indicated that individual variability in testosterone levels was primarily explained by environmental factors. Regarding genetic variation, univariate tests did not reveal any variants significantly associated with salivary testosterone after adjusting for false discovery rate. The top hit in males was rs10923844, an SNP of unknown function located downstream of HSD3B1 and HSD3B2. The top hits in females were two SNPs located upstream of ESR1 (rs3407085 and rs2295190). RF analysis, which reflects joint and conditional effects of multiple variants, indicated that genes involved in regulation of reproductive function, particularly LHCGR, are related to salivary testosterone levels in male infants, as are genes involved in cholesterol production, transport, and removal, while genes involved in estrogen signaling are related to salivary testosterone levels in female infants.
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The physicochemical property characterization of agar acetate.
Carbohydr Polym
PUBLISHED: 01-01-2014
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A series of agar acetates with different degree of substitution (DS) were prepared, and their properties were determined and analyzed. The results showed that the gelling temperature, the gel melting temperature, the gel strength, the gel hardness, the gel fracturability, the gel springiness and the solution apparent viscosity of agar acetates all decreased except that their gel cohesiveness increased with the increase of DS. The variation process of agar molecules in solution from coil to helix could be also observed by measuring solution optical rotation in a lower concentration at which even the solution could not form a gel. The gel skeleton structures of agar acetates were of porous network structures, and the pores became smaller and denser with the increase of DS. After acetylation, the water holding capacity of the agar was improved, but its thermal stability was lowered.
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Size-dependent cytotoxicity of nanocarbon blacks.
Int J Mol Sci
PUBLISHED: 09-22-2013
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In this study, we investigated the toxic effects of nanocarbon blacks (NCBs) with different sizes to mouse macrophage RAW264.7 cells. MTT and fluorescence-based LIVE assays demonstrated that NCBs uptake caused a size and dose-dependent growth inhibition to the cells. Optical microscopy observations and (99m)Tc radionuclide labeling techniques were used to investigate the cellular uptake of NCBs with different sizes qualitatively and quantitatively, respectively. Results showed that the cellular uptake amounts of NCBs increased with their increasing size. Large quantities of internal NCBs induced oxidative stress and nuclear damage in cells; these effects may be the critical factors involved in the cytotoxicity of NCBs. The implications associated with these findings are discussed.
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Identification and Characterization of Seven Glutathione S-Transferase Genes from Citrus Red Mite, Panonychus citri (McGregor).
Int J Mol Sci
PUBLISHED: 09-10-2013
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The citrus red mite, Panonychus citri (McGregor), is a global citrus pest, and has developed severe resistance to several types of acaricides. However, the molecular mechanisms of resistance in this mite remain unknown. In this study, seven full-length cDNAs encoding glutathione S-transferases (GSTs) genes were identified and characterized in P. citri. The effects of pyridaben and fenpropathrin exposure on the expression of these genes were also investigated. Phylogenetic analysis revealed that the seven GSTs genes in P. citri cloned in this study belong to three different cytosolic classes, including four in mu, two in delta and one in zeta. Among these seven GSTs genes, the relative expression level of PcGSTm1 was significantly higher in adult than in the other life stages (egg, larvae and nymph). Compared with the control, the mRNA levels of the seven GST genes did not change significantly following exposure to pyridaben at LC10. However, RT-qPCR results showed that, when exposed to LC10 of fenpropathrin, six GSTs gene (PcGSTm1, PcGSTm3, PcGSTm4, PcGSTd1, PcGSTd2 and PcGSTz1) transcripts increased in a time-dependent manner. This is the first insight into the molecular characteristics of GSTs gene cDNAs in P. citri. The elevated GSTs gene transcripts following exposure to fenpropathrin might be one of the mechanisms involved in detoxification of this acaricide.
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Genome-wide association analysis identifies 13 new risk loci for schizophrenia.
Stephan Ripke, Colm O'Dushlaine, Kimberly Chambert, Jennifer L Moran, Anna K Kähler, Susanne Akterin, Sarah E Bergen, Ann L Collins, James J Crowley, Menachem Fromer, Yunjung Kim, Sang Hong Lee, Patrik K E Magnusson, Nick Sanchez, Eli A Stahl, Stephanie Williams, Naomi R Wray, Kai Xia, Francesco Bettella, Anders D Borglum, Brendan K Bulik-Sullivan, Paul Cormican, Nick Craddock, Christiaan de Leeuw, Naser Durmishi, Michael Gill, Vera Golimbet, Marian L Hamshere, Peter Holmans, David M Hougaard, Kenneth S Kendler, Kuang Lin, Derek W Morris, Ole Mors, Preben B Mortensen, Benjamin M Neale, Francis A O'Neill, Michael J Owen, Milica Pejović Milovančević, Danielle Posthuma, John Powell, Alexander L Richards, Brien P Riley, Douglas Ruderfer, Dan Rujescu, Engilbert Sigurdsson, Teimuraz Silagadze, August B Smit, Hreinn Stefansson, Stacy Steinberg, Jaana Suvisaari, Sarah Tosato, Matthijs Verhage, James T Walters, , Douglas F Levinson, Pablo V Gejman, Claudine Laurent, Bryan J Mowry, Michael C O'Donovan, Ann E Pulver, Sibylle G Schwab, Dieter B Wildenauer, Frank Dudbridge, Jianxin Shi, Margot Albus, Madeline Alexander, Dominique Campion, David Cohen, Dimitris Dikeos, Jubao Duan, Peter Eichhammer, Stephanie Godard, Mark Hansen, F Bernard Lerer, Kung-Yee Liang, Wolfgang Maier, Jacques Mallet, Deborah A Nertney, Gerald Nestadt, Nadine Norton, George N Papadimitriou, Robert Ribble, Alan R Sanders, Jeremy M Silverman, Dermot Walsh, Nigel M Williams, Brandon Wormley, Maria J Arranz, Steven Bakker, Stephan Bender, Elvira Bramon, David Collier, Benedicto Crespo-Facorro, Jeremy Hall, Conrad Iyegbe, Assen Jablensky, René S Kahn, Luba Kalaydjieva, Stephen Lawrie, Cathryn M Lewis, Don H Linszen, Ignacio Mata, Andrew McIntosh, Robin M Murray, Roel A Ophoff, Jim van Os, Muriel Walshe, Matthias Weisbrod, Durk Wiersma, Peter Donnelly, Inês Barroso, Jenefer M Blackwell, Matthew A Brown, Juan P Casas, Aiden P Corvin, Panos Deloukas, Audrey Duncanson, Janusz Jankowski, Hugh S Markus, Christopher G Mathew, Colin N A Palmer, Robert Plomin, Anna Rautanen, Stephen J Sawcer, Richard C Trembath, Ananth C Viswanathan, Nicholas W Wood, Chris C A Spencer, Gavin Band, Celine Bellenguez, Colin Freeman, Garrett Hellenthal, Eleni Giannoulatou, Matti Pirinen, Richard D Pearson, Amy Strange, Zhan Su, Damjan Vukcevic, Cordelia Langford, Sarah E Hunt, Sarah Edkins, Rhian Gwilliam, Hannah Blackburn, Suzannah J Bumpstead, Serge Dronov, Matthew Gillman, Emma Gray, Naomi Hammond, Alagurevathi Jayakumar, Owen T McCann, Jennifer Liddle, Simon C Potter, Radhi Ravindrarajah, Michelle Ricketts, Avazeh Tashakkori-Ghanbaria, Matthew J Waller, Paul Weston, Sara Widaa, Pamela Whittaker, Mark I McCarthy, Kari Stefansson, Edward Scolnick, Shaun Purcell, Steven A McCarroll, Pamela Sklar, Christina M Hultman, Patrick F Sullivan.
Nat. Genet.
PUBLISHED: 08-01-2013
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Schizophrenia is an idiopathic mental disorder with a heritable component and a substantial public health impact. We conducted a multi-stage genome-wide association study (GWAS) for schizophrenia beginning with a Swedish national sample (5,001 cases and 6,243 controls) followed by meta-analysis with previous schizophrenia GWAS (8,832 cases and 12,067 controls) and finally by replication of SNPs in 168 genomic regions in independent samples (7,413 cases, 19,762 controls and 581 parent-offspring trios). We identified 22 loci associated at genome-wide significance; 13 of these are new, and 1 was previously implicated in bipolar disorder. Examination of candidate genes at these loci suggests the involvement of neuronal calcium signaling. We estimate that 8,300 independent, mostly common SNPs (95% credible interval of 6,300-10,200 SNPs) contribute to risk for schizophrenia and that these collectively account for at least 32% of the variance in liability. Common genetic variation has an important role in the etiology of schizophrenia, and larger studies will allow more detailed understanding of this disorder.
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Peptide fragments of human serum albumin as novel renal targeting carriers.
Int J Pharm
PUBLISHED: 07-29-2013
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To develop the proper renal targeting carrier for clinical therapy, human serum albumin, as starting material, was firstly cleaved into albumin fragments and Superdex 75 and CM-Sepharose FF were used to separate and purify the degradation products. Consequently, three peptide fragments (PFs) with certain sequence named PF-A1-123, PF-A124-298 and PF-A299-585 were obtained as candidates of renal targeting carrier. Then, cytotoxicity and cellular uptake of three PFs was studied preliminarily. The results showed that three PFs had no adverse effects on the HeLa and MDCK cell even up to 5.00mg/mL and PF-A299-585 exhibited highest affinity to MDCK cells. After that, we found that PFs selectively accumulated in the kidneys, especially in the renal tubules after intravenous injection in mice by optical imaging study. Finally, Tissues distribution in vivo was utilized to verify the renal targeting profiles of PFs. Three PFs exhibited renal accumulation characteristics. In particular, about 40% injected doses of PF-A299-585 were specifically distributed into kidneys for 1h. The mean area under the curve (AUC) of PF-A299-585 in kidneys increased 13 times compared with those of PF-A1-123 and PF-A124-298. Therefore, PFs can be applied as prospective carriers for renal targeting and PF-A299-585 may be the optimal carrier.
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Functional analysis of the distal region of the third intracellular loop of PROKR2.
Biochem. Biophys. Res. Commun.
PUBLISHED: 07-29-2013
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Mutations in the G-protein-coupled receptor PROKR2 have been identified in patients with idiopathic hypogonadotropic hypogonadism (IHH) and Kallmann syndrome (KS) manifesting with delayed puberty and infertility. Recently, the homozygous mutation V274D was identified in a man displaying KS with an apparent reversal of hypogonadism. The affected amino acid, valine 274, is located at the junction region of the third intracellular loop (IL3) and the sixth transmembrane domain (TM6). In this study, we first studied the effect of V274D and related mutations (V274A, V274T, and V274R) on the signaling activity and cell surface expression of PROKR2. Our data indicate that a charged amino acid substitution at residue 274 of PROKR2 results in low cell surface expression and loss-of-function. Furthermore, we studied the effects of two clusters of basic amino acids located at the proximal region of Val274 on the cell surface expression and function of PROKR2. The deletion of RRK (270-272) resulted in undetectable cell surface expression, whereas RKR (264-266)-deleted PROKR2 was expressed normally on the cell surface but showed loss-of-function due to a deficiency in G-protein coupling. Our data indicate that the distal region of the IL3 of PROKR2 may differentially influence receptor trafficking and G-protein coupling.
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[Treatment of refractory lateral epicondylitis with modified Nirschl surgical technique].
Zhongguo Gu Shang
PUBLISHED: 06-26-2013
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To investigate the clinical effects of modified Nirschl surgical techniique in treating refractory lateral epicondylitis.
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Improved quality control method for prescriptions of Polygonum capitatum through simultaneous determination of nine major constituents by HPLC coupled with triple quadruple mass spectrometry.
Molecules
PUBLISHED: 06-13-2013
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As a traditional Miao-nationality medicinal plant, Polygonum capitatum has been used in clinical practice for several thousand years. Its prescriptions, including three dosage forms: granules, capsule and tablet are known by the brand name Relinqing® and have played an indispensable role in the treatment of urinary system infection, pyelonephritis and kidney stones. However, no study about the comprehensive quality evaluation of Relinqing® has been reported. In the present paper, a method for the simultaneous determination of nine major compounds in three dosage forms of Relinqing® using HPLC coupled with triple quadrupole mass spectrometry (HPLC-QQQ MS) was established to comprehensively evaluate their quality. The nine compounds, including four phenolic acids, four flavonoids and a lignin, were analyzed with acceptable linear regression relationship (r², 0.9923-0.9992), precision (RSD, 1.25%-2.78%), repeatability (RSD, 2.05%%-3.47%), stability (RSD, 1.84%-3.72%) and recovery (93.60%-108.54%, RSD ? 3.67%). The present study fills the gap in the multivariate quality control of Relinqing® and provides a valuable reference for quality standards and dosage reforming of this traditional Chinese medicine.
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Impact of Pre-existing Type-2 Diabetes on Patient Outcomes After Radical Resection for Gastric Cancer: A Retrospective Cohort Study.
Dig. Dis. Sci.
PUBLISHED: 06-12-2013
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The aim of this study was to evaluate the impact of pre-existing type-2 diabetes on postoperative recovery and prognosis in gastric cancer (GC) patients who underwent radical gastrectomy.
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Transglutaminase 6 interacts with polyQ proteins and promotes the formation of polyQ aggregates.
Biochem. Biophys. Res. Commun.
PUBLISHED: 06-11-2013
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A common feature of polyglutamine (polyQ) diseases is the presence of aggregates in neuronal cells caused by expanded polyglutamine tracts. PolyQ proteins are the substrates of transglutaminase 2, and the increased activity of transglutaminase in polyQ diseases suggests that transglutaminase may be directly involved in the formation of the aggregates. We previously identified the transglutaminase 6 gene to be causative of spinocerebellar ataxia type 35 (SCA35), and we found that SCA35-associated mutants exhibited reduced transglutaminase activity. Here we report that transglutaminase 6 interacts and co-localizes with both normal and expanded polyQ proteins in HEK293 cells. Moreover, the overexpression of transglutaminase 6 promotes the formation of polyQ aggregates and the conversion of soluble polyQ into insoluble polyQ aggregates. However, SCA35-associated mutants do not affect their interactions with polyQ proteins. These data suggest that transglutaminase 6 could be involved in polyQ diseases and there may exist a common pathological link between polyQ associated SCA and SCA35.
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[Treatment of anteromedial coronoid facet fractures with open reduction and internal fixation through anteromedial approach].
Zhongguo Gu Shang
PUBLISHED: 05-18-2013
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To investigate the approach of open reduction and internal fixation for the treatment of anteromedial coronoid facet fractures and associated injury.
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Identification of seven loci affecting mean telomere length and their association with disease.
Veryan Codd, Christopher P Nelson, Eva Albrecht, Massimo Mangino, Joris Deelen, Jessica L Buxton, Jouke Jan Hottenga, Krista Fischer, Tonu Esko, Ida Surakka, Linda Broer, Dale R Nyholt, Irene Mateo Leach, Perttu Salo, Sara Hägg, Mary K Matthews, Jutta Palmen, Giuseppe D Norata, Paul F O'Reilly, Danish Saleheen, Najaf Amin, Anthony J Balmforth, Marian Beekman, Rudolf A de Boer, Stefan Böhringer, Peter S Braund, Paul R Burton, Anton J M de Craen, Matthew Denniff, Yanbin Dong, Konstantinos Douroudis, Elena Dubinina, Johan G Eriksson, Katia Garlaschelli, Dehuang Guo, Anna-Liisa Hartikainen, Anjali K Henders, Jeanine J Houwing-Duistermaat, Laura Kananen, Lennart C Karssen, Johannes Kettunen, Norman Klopp, Vasiliki Lagou, Elisabeth M van Leeuwen, Pamela A Madden, Reedik Mägi, Patrik K E Magnusson, Satu Mannisto, Mark I McCarthy, Sarah E Medland, Evelin Mihailov, Grant W Montgomery, Ben A Oostra, Aarno Palotie, Annette Peters, Helen Pollard, Anneli Pouta, Inga Prokopenko, Samuli Ripatti, Veikko Salomaa, H Eka D Suchiman, Ana M Valdes, Niek Verweij, Ana Viñuela, Xiaoling Wang, H-Erich Wichmann, Elisabeth Widén, Gonneke Willemsen, Margaret J Wright, Kai Xia, Xiangjun Xiao, Dirk J van Veldhuisen, Alberico L Catapano, Martin D Tobin, Alistair S Hall, Alexandra I F Blakemore, Wiek H van Gilst, Haidong Zhu, Cardiogram Consortium, Jeanette Erdmann, Muredach P Reilly, Sekar Kathiresan, Heribert Schunkert, Philippa J Talmud, Nancy L Pedersen, Markus Perola, Willem Ouwehand, Jaakko Kaprio, Nicholas G Martin, Cornelia M van Duijn, Iiris Hovatta, Christian Gieger, Andres Metspalu, Dorret I Boomsma, Marjo-Riitta Järvelin, P Eline Slagboom, John R Thompson, Tim D Spector, Pim van der Harst, Nilesh J Samani.
Nat. Genet.
PUBLISHED: 03-29-2013
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Interindividual variation in mean leukocyte telomere length (LTL) is associated with cancer and several age-associated diseases. We report here a genome-wide meta-analysis of 37,684 individuals with replication of selected variants in an additional 10,739 individuals. We identified seven loci, including five new loci, associated with mean LTL (P < 5 × 10(-8)). Five of the loci contain candidate genes (TERC, TERT, NAF1, OBFC1 and RTEL1) that are known to be involved in telomere biology. Lead SNPs at two loci (TERC and TERT) associate with several cancers and other diseases, including idiopathic pulmonary fibrosis. Moreover, a genetic risk score analysis combining lead variants at all 7 loci in 22,233 coronary artery disease cases and 64,762 controls showed an association of the alleles associated with shorter LTL with increased risk of coronary artery disease (21% (95% confidence interval, 5-35%) per standard deviation in LTL, P = 0.014). Our findings support a causal role of telomere-length variation in some age-related diseases.
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Family history and risk of ductal carcinoma in situ and triple negative breast cancer in a Han Chinese population: a case-control study.
World J Surg Oncol
PUBLISHED: 03-13-2013
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The association between family history and risk of triple negative breast cancer and ductal carcinoma in situ (DCIS) has not been well investigated, especially in Asian populations. We investigated the association between family history and risk of DCIS or triple negative breast cancer in a Han Chinese population.
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Associations of prenatal exposure to phenols with birth outcomes.
Environ. Pollut.
PUBLISHED: 03-09-2013
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Many phenols are known to mimic or antagonize hormonal activities and may adversely affect fetal growth. A study of 567 pregnant women was conducted to investigate the relationship between prenatal phenol exposure and birth outcomes, including birth weight, length, and gestational age. We measured the concentrations of bisphenol A, benzophenone-3, 4-n-octylphenol and 4-n-nonylphenol in maternal urine and examine their association with birth outcomes. Categories of urinary benzophenone-3 concentration were associated with decreased gestational age in all infants (p for trend = 0.03). Between middle and low exposure groups, we also found bisphenol A was negatively associated with gestational duration (?adjusted = -0.48 week; 95% confidence interval: -0.91, -0.05). After stratification by gender, we found the consistent results in infant boys with those in all infants, but we did not observe significant association for girls. In conclusion, we found prenatal phenol exposure was sex-specifically related to birth outcomes.
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Efficacy of adjuvant XELOX and FOLFOX6 chemotherapy after D2 dissection for gastric cancer.
World J. Gastroenterol.
PUBLISHED: 01-30-2013
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To compare the efficacy of capecitabine and oxaliplatin (XELOX) with 5-fluorouracil, folinic acid and oxaliplatin (FOLFOX6) in gastric cancer patients after D2 dissection.
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Genetic polymorphism in hOGG1 is associated with triple-negative breast cancer risk in Chinese Han women.
Breast
PUBLISHED: 01-29-2013
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8-hydroxy-2-deoxyguanine (8-OHdG), a typical product of oxidative stress-induced DNA damage, can cause a G-T transversion during DNA replication if it is not removed. Human 8-oxoguanine glycosylase 1 (hOGG1), a key DNA repair gene, recognizes and excises 8-OHdG from damaged DNA accurately; however, a c.977C>G (Ser326Cys) polymorphism in hOGG1 can inhibit the genes ability to remove 8-OHdG. The aim of present study was to investigate the association between the c.977C>G polymorphism in hOGG1 and the risk of breast cancer in Chinese Han women. We used high-resolution melting and sequencing to analyze the genotypes of 630 patients with sporadic breast cancer patients and 777 healthy controls. We also performed risk-stratified subgroup analyses to determine the association between the c.977C>G polymorphism and other characteristics of breast cancer subgroups. Breast cancer patients and healthy controls did not have significantly different of c.977C/G genotypes (odds ratio [OR] = 1.10, 95% confidence interval [CI] = 0.82-1.49, p = 0.57) and c.977G/G genotypes (OR = 1.34, 95% CI = 0.97-1.84, p = 0.09). However, the c.977G/G genotype was especially prevalent in breast cancer patients who were younger than 55 years (OR = 1.58, 95% CI = 1.05-2.39, p = 0.04), were premenopausal status (OR = 1.87, 95% CI = 1.14-3.06, p = 0.02), had triple-negative disease (OR = 2.14, 95% CI = 1.06-4.29, p = 0.04), or p53-positive disease (OR = 1.56, 95% CI = 1.14-2.12, p = 0.005). These findings suggest that the c.977C>G polymorphism in hOGG1 is associated with an increased risk of breast cancer in Chinese Han women who are younger than 55 years, premenopausal, triple-negative, or p53-positive subgroups.
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Prostaglandin E2 promotes liver cancer cell growth by the upregulation of FUSE-binding protein 1 expression.
Int. J. Oncol.
PUBLISHED: 01-18-2013
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Liver cancer is a common human cancer with a high mortality rate and currently there is no effective chemoprevention or systematic treatment. Recent evidence suggests that prostaglandin E(2) (PGE(2)) plays an important role in the occurrence and development of liver cancer. However, the mechanisms through which PGE(2) promotes liver cancer cell growth are not yet fully understood. It has been reported that the increased expression of FUSE-binding protein 1 (FBP1) significantly induces the proliferation of liver cancer cells. In this study, we report that PGE(2) promotes liver cancer cell growth by the upregulation of FBP1 protein expression. Treatment with PGE2 and the E prostanoid 3 (EP3) receptor agonist, sulprostone, resulted in the time-dependent increase in FBP1 protein expression; sulprostone increased the viability of the liver cancer cells. The protein kinase A (PKA) inhibitor, H89, and the adenylate cyclase (AC) inhibitor, SQ22536, inhibited the cell viability accelerated by sulprostone. By contrast, the Gi subunit inhibitor, pertussis toxin (PTX), exhibited no significant effect. Treatment with PGE(2) and sulprostone caused a decrease in JTV1 protein expression, blocked the binding of JTV1 with FBP1, which served as a mechanism for FBP1 degradation, leading to the decreased ubiquitination of FBP1 and the increase in FBP1 protein expression. Furthermore, H89 and SQ22536 prevented the above effects of JTV1 and FBP1 induced by PGE(2) and sulprostone. These findings indicate that the EP3 receptor activated by PGE(2) may couple to Gs protein and activate cyclic AMP (cAMP)-PKA, downregulating the levels of JTV1 protein, consequently inhibiting the ubiquitination of FBP1 and increasing FBP1 protein expression, thus promoting liver cancer cell growth. These observations provide new insights into the mechanisms through which PGE(2) promotes cancer cell growth.
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Hydrogen Peroxide Is a Second Messenger in the Salicylic Acid-Triggered Adventitious Rooting Process in Mung Bean Seedlings.
PLoS ONE
PUBLISHED: 01-01-2013
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In plants, salicylic acid (SA) is a signaling molecule that regulates disease resistance responses, such as systemic acquired resistance (SAR) and hypertensive response (HR). SA has been implicated as participating in various biotic and abiotic stresses. This study was conducted to investigate the role of SA in adventitious root formation (ARF) in mung bean (Phaseolus radiatus L) hypocotyl cuttings. We observed that hypocotyl treatment with SA could significantly promote the adventitious root formation, and its effects were dose and time dependent. Explants treated with SA displayed a 130% increase in adventitious root number compared with control seedlings. The role of SA in mung bean hypocotyl ARF as well as its interaction with hydrogen peroxide (H2O2) were also elucidated. Pretreatment of mung bean explants with N, N-dimethylthiourea (DMTU), a scavenger for H2O2, resulted in a significant reduction of SA-induced ARF. Diphenyleneiodonium (DPI), a specific inhibitor of membrane-linked NADPH oxidase, also inhibited the effect of adventitious rooting triggered by SA treatment. The determination of the endogenous H2O2 level indicated that the seedlings treated with SA could induce H2O2 accumulation compared with the control treatment. Our results revealed a distinctive role of SA in the promotion of adventitious rooting via the process of H2O2 accumulation. This conclusion was further supported by antioxidant enzyme activity assays. Based on these results, we conclude that the accumulation of free H2O2 might be a downstream event in response to SA-triggered adventitious root formation in mung bean seedlings.
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No association between TGFB1 polymorphisms and late radiotherapy toxicity: a meta-analysis.
PLoS ONE
PUBLISHED: 01-01-2013
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Transforming growth factor-beta 1 (TGF-?1) protein may be multifunctional and related to the development of fibrosis, induction of apoptosis, extracellular signaling and inhibition of proliferation in response to radiation-induced DNA damage. Several studies have investigated associations between single nucleotide polymorphisms (SNPs) in the TGFB1 gene and risk of late radiation-induced injury of normal tissue, but the conclusions remain controversial.
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Low serum creatine kinase levels in breast cancer patients: a case-control study.
PLoS ONE
PUBLISHED: 01-01-2013
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Previous studies provide an ambiguous picture of creatine kinase (CK) expression and activities in malignancy. The aim of this study was to investigate the role of serum CK level in breast cancer patients.
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seeQTL: a searchable database for human eQTLs.
Bioinformatics
PUBLISHED: 12-13-2011
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seeQTL is a comprehensive and versatile eQTL database, including various eQTL studies and a meta-analysis of HapMap eQTL information. The database presents eQTL association results in a convenient browser, using both segmented local-association plots and genome-wide Manhattan plots.
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Preparation and immunogenicity of tag-free recombinant human eppin.
Asian J. Androl.
PUBLISHED: 09-05-2011
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Human epididymal protease inhibitor (eppin) may be effective as a male contraceptive vaccine. In a number of studies, eppin with an engineered His(6)-tag has been produced using prokaryotic expression systems. For production of pharmaceutical-grade proteins for human use, however, the His(6)-tag must be removed. This study describes a method for producing recombinant human eppin without a His(6)-tag. We constructed plasmid pET28a (+)-His(6)-tobacco etch virus (TEV)-eppin for expression in Escherichia coli. After purification and refolding, the fusion protein His(6)-TEV-eppin was digested with TEV protease to remove the His(6)-tag and was further purified by NTA-Ni(2+) affinity chromatography. Using this procedure, 2 mg of eppin without a His(6)-tag was isolated from 1 l of culture with a purity of >95%. The immunogenicity of the eppin was characterized using male Balb/c mice.
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A powerful and flexible approach to the analysis of RNA sequence count data.
Bioinformatics
PUBLISHED: 08-02-2011
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A number of penalization and shrinkage approaches have been proposed for the analysis of microarray gene expression data. Similar techniques are now routinely applied to RNA sequence transcriptional count data, although the value of such shrinkage has not been conclusively established. If penalization is desired, the explicit modeling of mean-variance relationships provides a flexible testing regimen that borrows information across genes, while easily incorporating design effects and additional covariates.
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The association of APE1 -656T > G and 1349 T > G polymorphisms and cancer risk: a meta-analysis based on 37 case-control studies.
BMC Cancer
PUBLISHED: 06-30-2011
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APE1 (apurinic/apyrimidinic endonuclease 1) is an important DNA repair protein in the base excision repair pathway. Polymorphisms in APE1 have been implicated in susceptibility to cancer; however, results from the published studies remained inconclusive. The objective of this study was to conduct a meta-analysis investigating the association between polymorphisms in APE1 and the risk for cancer.
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Functional characterization of TRICHOMELESS2, a new single-repeat R3 MYB transcription factor in the regulation of trichome patterning in Arabidopsis.
BMC Plant Biol.
PUBLISHED: 05-08-2011
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Single-repeat R3 MYB transcription factors (single-repeat MYBs) play important roles in controlling trichome patterning in Arabidopsis. It was proposed that single-repeat MYBs negatively regulate trichome formation by competing with GLABRA1 (GL1) for binding GLABRA3/ENHANCER OF GLABRA3 (GL3/EGL3), thus inhibiting the formation of activator complex TTG1(TRANSPARENT TESTA GLABRA1)-GL3/EGL3-GL1 that is required for the activation of GLABRA2 (GL2), whose product is a positive regulator of trichome formation. Previously we identified a novel single-repeat MYB transcription factor, TRICHOMELESS1 (TCL1), which negatively regulates trichome formation on the inflorescence stems and pedicels by directly suppressing the expression of GL1.
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Fasudil hydrochloride hydrate, a Rho-kinase inhibitor, suppresses high glucose-induced proliferation and collagen synthesis in rat cardiac fibroblasts.
Clin. Exp. Pharmacol. Physiol.
PUBLISHED: 04-05-2011
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1. Hyperglycaemia promotes the proliferation of cardiac fibroblasts (CFs) and collagen synthesis in CFs. The objectives of the present study were to determine the effects of fasudil hydrochloride hydrate, a Rho-kinase (ROCK) inhibitor, on high glucose (HG)-induced proliferation of CFs and collagen production in rat CFs and to investigate the molecular mechanism of action of fasudil. 2. Rat CFs were cultured in Dulbeccos modified Eagles medium, supplemented with 5.5 or 25 mmol/L d-glucose or 5.5 mmol/L d-glucose + 19.5 mmol/L mannose, in the presence of absence of fasudil (50 or 100 ?mol/L). Proliferation was measured by the 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, whereas the production of Type I collagen was evaluated using ELISA and the expression of ROCK1, c-Jun N-terminal kinase (JNK) and Type I procollagen mRNA was determined by reverse transcription-polymerase chain reaction. Intracellular Type I procollagen protein levels were evaluated using immunocytochemistry. Western blot analysis was used to evaluate the phosphorylation of myosin phosphatase target subunit 1 (MYPT1), JNK and Smad2/3, as well as c-jun protein levels. 3. Both concentrations of fasudil effectively inhibited HG (25 mmol/L d-glucose)-induced increases in the proliferation of CFs and collagen synthesis, concomitant with suppression of HG-induced upregulation of ROCK1 and JNK mRNA expression and c-jun protein levels, as well as the phosphorylation of MYPT1, JNK and Smad2/3. 4. These data suggest that ROCK activation is essential for the proliferation of CFs and collagen synthesis induced by HG. Fasudil suppressed HG-induced increases in the proliferation of CFs and collagen synthesis, which may be associated with inhibition of the JNK and transforming growth factor ?/Smad pathways. The results of the present study indicate that inhibition of ROCK may be a novel therapeutic target for the prevention of diabetic cardiac fibrosis.
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Dual functions of Tet1 in transcriptional regulation in mouse embryonic stem cells.
Nature
PUBLISHED: 02-14-2011
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Epigenetic modification of the mammalian genome by DNA methylation (5-methylcytosine) has a profound impact on chromatin structure, gene expression and maintenance of cellular identity. The recent demonstration that members of the Ten-eleven translocation (Tet) family of proteins can convert 5-methylcytosine to 5-hydroxymethylcytosine raised the possibility that Tet proteins are capable of establishing a distinct epigenetic state. We have recently demonstrated that Tet1 is specifically expressed in murine embryonic stem (ES) cells and is required for ES cell maintenance. Using chromatin immunoprecipitation coupled with high-throughput DNA sequencing, here we show in mouse ES cells that Tet1 is preferentially bound to CpG-rich sequences at promoters of both transcriptionally active and Polycomb-repressed genes. Despite an increase in levels of DNA methylation at many Tet1-binding sites, Tet1 depletion does not lead to downregulation of all the Tet1 targets. Interestingly, although Tet1-mediated promoter hypomethylation is required for maintaining the expression of a group of transcriptionally active genes, it is also involved in repression of Polycomb-targeted developmental regulators. Tet1 contributes to silencing of this group of genes by facilitating recruitment of PRC2 to CpG-rich gene promoters. Thus, our study not only establishes a role for Tet1 in modulating DNA methylation levels at CpG-rich promoters, but also reveals a dual function of Tet1 in promoting transcription of pluripotency factors as well as participating in the repression of Polycomb-targeted developmental regulators.
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Toward accurate prediction of pKa values for internal protein residues: the importance of conformational relaxation and desolvation energy.
Proteins
PUBLISHED: 01-11-2011
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Proton uptake or release controls many important biological processes, such as energy transduction, virus replication, and catalysis. Accurate pK(a) prediction informs about proton pathways, thereby revealing detailed acid-base mechanisms. Physics-based methods in the framework of molecular dynamics simulations not only offer pK(a) predictions but also inform about the physical origins of pK(a) shifts and provide details of ionization-induced conformational relaxation and large-scale transitions. One such method is the recently developed continuous constant pH molecular dynamics (CPHMD) method, which has been shown to be an accurate and robust pK(a) prediction tool for naturally occurring titratable residues. To further examine the accuracy and limitations of CPHMD, we blindly predicted the pK(a) values for 87 titratable residues introduced in various hydrophobic regions of staphylococcal nuclease and variants. The predictions gave a root-mean-square deviation of 1.69 pK units from experiment, and there were only two pK(a)s with errors greater than 3.5 pK units. Analysis of the conformational fluctuation of titrating side-chains in the context of the errors of calculated pK(a) values indicate that explicit treatment of conformational flexibility and the associated dielectric relaxation gives CPHMD a distinct advantage. Analysis of the sources of errors suggests that more accurate pK(a) predictions can be obtained for the most deeply buried residues by improving the accuracy in calculating desolvation energies. Furthermore, it is found that the generalized Born implicit-solvent model underlying the current CPHMD implementation slightly distorts the local conformational environment such that the inclusion of an explicit-solvent representation may offer improvement of accuracy.
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Smad2 mediates Activin/Nodal signaling in mesendoderm differentiation of mouse embryonic stem cells.
Cell Res.
PUBLISHED: 11-16-2010
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Although Activin/Nodal signaling regulates pluripotency of human embryonic stem (ES) cells, how this signaling acts in mouse ES cells remains largely unclear. To investigate this, we confirmed that mouse ES cells possess active Smad2-mediated Activin/Nodal signaling and found that Smad2-mediated Activin/Nodal signaling is dispensable for self-renewal maintenance but is required for proper differentiation toward the mesendoderm lineage. To gain insights into the underlying mechanisms, Smad2-associated genes were identified by genome-wide chromatin immunoprecipitation-chip analysis. The results showed that there is a transcriptional correlation between Smad2 binding and Activin/Nodal signaling modulation, and that the development-related genes were enriched among the Smad2-bound targets. We further identified Tapbp as a key player in mesendoderm differentiation of mouse ES cells acting downstream of the Activin/Nodal-Smad2 pathway. Taken together, our findings suggest that Smad2-mediated Activin/Nodal signaling orchestrates mesendoderm lineage commitment of mouse ES cells through direct modulation of corresponding developmental regulator expression.
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BCL2 Ala43Thr is a functional variant associated with protection against azoospermia in a Han-Chinese population.
Biol. Reprod.
PUBLISHED: 07-07-2010
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Apoptosis is very common during various stages of mammalian germ cell development and differentiation, and the BCL2 gene is one of the most important apoptotic regulators. Although its genetic variants are reported to be involved in cancers and autoimmune diseases, little information is available regarding BCL2 polymorphisms in male spermatogenesis. In the present study, single nucleotide polymorphisms (SNPs) in coding regions of the BCL2 gene were examined in a hospital-based, case-control study including 198 infertile patients with idiopathic azoospermia and 183 fertile controls. Subsequently, a functional study was conducted for comparison of paclitaxel-induced cytotoxicity and apoptosis between the BCL2 variant and the wild type in vitro. Three SNPs were found in exon 2--A21G (rs1801018), G127A (rs1800477), and C300T (rs61733416)--with the latter first reported in the Han-Chinese population. The frequency of G127A (GA+AA) genotype was significantly lower in azoospermic, infertile men compared to the age-matched controls (P = 0.01). This genotype may confer a lower risk of azoospermia (adjusted odd ratio [OR] = 0.448, 95% confidence interval = 0.226-0.889). In addition, HeLa cells expressing the BCL2 Ala43Thr (G127A), similar to the control cells, were more sensitive to paclitaxel-induced cytotoxicity and apoptosis than those expressing wild-type BCL2. Consistently, the cleaved PARP and p-BCL2 proteins were subsequently increased after paclitaxel treatment, as also predicted by the bioinformatics analysis. Considering the decreased antiapoptotic function of BCL2, these results suggest that the Ala43Thr variant is associated with protection against azoospermia in the Han-Chinese population.
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Butyrate promotes induced pluripotent stem cell generation.
J. Biol. Chem.
PUBLISHED: 06-16-2010
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Recent studies have demonstrated that embryonic stem cell-like induced pluripotent stem (iPS) cells can be generated by enforced expression of defined transcription factors. The fact that cell fate change is accompanied by changes in epigenetic modifications prompted us to investigate whether chemicals known to modulate epigenetic regulators are capable of enhancing the efficiency of iPS cell generation. Here, we report that butyrate, a natural small fatty acid and histone deacetylase inhibitor, significantly increases the efficiency of mouse iPS cell generation using the transcription factors Oct4, Sox2, Klf4, and c-Myc. We show that butyrate not only changes the reprogramming dynamics, but also increases the ratio of iPS cell colonies to total colonies by reducing the frequency of partially reprogrammed cells and transformed cells. Detailed analysis reveals that the effect of butyrate on reprogramming appears to be mediated by c-Myc and occurs during an early stage of reprogramming. Genome-wide gene expression analysis reveals up-regulation of ES cell-enriched genes when mouse embryonic fibroblasts are treated with butyrate during reprogramming. Thus, our study identifies butyrate as a chemical factor capable of promoting iPS cell generation.
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[Ala499Val (C > T) and Lys939Gln (A > C) polymorphisms of the XPC gene: their correlation with male infertility].
Zhonghua Nan Ke Xue
PUBLISHED: 04-08-2010
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The Ala499Val (C > T) and Lys939Gln (A > C) of the XPC gene are two potentially functional nonsynonymous polymorphisms, which affect the rate of DNA repair and might change XPC production and activity. This study aimed to explore the distribution of these two polymorphisms in the Chinese Han population and their relationship with male infertility.
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[Immunoadjuvant effect of Hsp70L1 in tumor vaccine].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
PUBLISHED: 04-07-2010
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To explore the immune enhancement of Hsp70L1 in the tumor cell vaccines.
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Genome-wide mapping of SMAD target genes reveals the role of BMP signaling in embryonic stem cell fate determination.
Genome Res.
PUBLISHED: 11-19-2009
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Embryonic stem (ES) cells are under precise control of both intrinsic self-renewal gene regulatory network and extrinsic growth factor-triggered signaling cascades. How external signaling pathways connect to core self-renewal transcriptional circuits is largely unknown. To probe this, we chose BMP signaling, which is previously recognized as a master control for both self-renewal and lineage commitment of murine ES cells. Here, we mapped target gene promoter occupancy of SMAD1/5 and SMAD4 on a genome-wide scale and found that they associate with a large group of developmental regulators that are enriched for H3K27 trimethylation and H3K4 trimethylation bivalent marks and are repressed in the self-renewing state, whereas they are rapidly induced upon differentiation. Smad knockdown experiments further indicate that SMAD-mediated BMP signaling is largely required for differentiation-related processes rather than directly influencing self-renewal. Among the SMAD-associated genes, we further identified Dpysl2 (previously known as Crmp2) and the H3K27 demethylase Kdm6b (previously known as Jmjd3) as BMP4-modulated early neural differentiation regulators. Combined with computational analysis, our results suggest that SMAD-mediated BMP signaling balances self-renewal versus differentiation by modulating a set of developmental regulators.
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Fenvalerate inhibits the growth of primary cultured rat preantral ovarian follicles.
Toxicology
PUBLISHED: 08-18-2009
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Fenvalerate is a widely used synthetic pyrethroid insecticide and is reported to disrupt reproductive function in humans and animals. However, little is known about its influence on follicular development. In this study, rat preantral follicles were primary cultured to investigate the effects of fenvalerate on follicular survival rate, morphological change, steroid hormone levels and steroidogenesis related gene mRNA expression. Follicles were cultured with 0, 1, 5 and 25 micromol/L fenvalerate for 72 h. And then the morphous was assessed by conventional light microscopy, steroid hormones were measured by RIA, and the expressions of steroidogenic acute regulatory protein (StAR) and cytochrome P450 side-chain cleavage enzyme (P450scc) were monitored by real-time quantitative PCR analysis. Results showed that fenvalerate inhibited the augmentation of follicular diameters but did not have detectable effects on follicular survival rates. The level of steroid hormones, such as progesterone, testosterone and estradiol, was inhibited. The inhibition might be due to the decreased expression levels of StAR and P450scc. These results suggested that fenvalerate restrained the follicular growth, and inhibited steroidogenesis by reducing StAR and P450scc gene expression, which might further contribute to the fenvalerate-induced reproductive dysfunction.
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Genome-wide uH2A localization analysis highlights Bmi1-dependent deposition of the mark at repressed genes.
PLoS Genet.
PUBLISHED: 05-05-2009
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Polycomb group (PcG) proteins control organism development by regulating the expression of developmental genes. Transcriptional regulation by PcG proteins is achieved, at least partly, through the PRC2-mediated methylation on lysine 27 of histone H3 (H3K27) and PRC1-mediated ubiquitylation on lysine 119 of histone H2A (uH2A). As an integral component of PRC1, Bmi1 has been demonstrated to be critical for H2A ubiquitylation. Although recent studies have revealed the genome-wide binding patterns of some of the PRC1 and PRC2 components, as well as the H3K27me3 mark, there have been no reports describing genome-wide localization of uH2A. Using the recently developed ChIP-Seq technology, here, we report genome-wide localization of the Bmi1-dependent uH2A mark in MEF cells. Gene promoter averaging analysis indicates a peak of uH2A just inside the transcription start site (TSS) of well-annotated genes. This peak is enriched at promoters containing the H3K27me3 mark and represents the least expressed genes in WT MEF cells. In addition, peak finding reveals regions of local uH2A enrichment throughout the mouse genome, including almost 700 gene promoters. Genes with promoter peaks of uH2A exhibit lower-level expression when compared to genes that do not contain promoter peaks of uH2A. Moreover, we demonstrate that genes with uH2A peaks have increased expression upon Bmi1 knockout. Importantly, local enrichment of uH2A is not limited to regions containing the H3K27me3 mark. We describe the enrichment of H2A ubiquitylation at high-density CpG promoters and provide evidence to suggest that DNA methylation may be linked to uH2A at these regions. Thus, our work not only reveals Bmi1-dependent H2A ubiquitylation, but also suggests that uH2A targeting in differentiated cells may employ a different mechanism from that in ES cells.
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Low molecular weight hydroxyethyl chitosan-prednisolone conjugate for renal targeting therapy: synthesis, characterization and in vivo studies.
Theranostics
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To further evaluate the potential renal targeting profile of low molecular weight hydroxyethyl chitosan (LMWHC) we developed before, prednisolone (Pre) was conjugated with LMWHC by EDC/NHS chemistry to improve the therapeutic effect of glucocorticoids in vivo. The conjugate was denoted as LMWHC-Pre. The prednisolone content of the conjugate was determined by reversed-phase high-performance liquid chromatography (HPLC) with Kromasil C18 column. The results showed that the average coupling degree of prednisolone to LMWHC was 76.7±3.2 ?g·mg(-1). The stability and physicochemical characterization of LMWHC-Pre under various conditions were also investigated. To study the fate of LMWHC-Pre after intravenous (i.v.) administration, fluorescein isothiocyanate (FITC) was coupled to the conjugate to explore the renal targeting efficacy. The in vivo results showed that significant amount of the conjugate was accumulated into the kidneys while negligible signal could be detected when the mixture of FITC-LMWHC and prednisolone was co-administered. The preliminary pharmacodynamics study of LMWHC-Pre showed that the conjugate could effectively alleviate the nephrotic syndrome of rats induced by minimal change nephrosis (MCN) model. Toxicity study also revealed that there was little glucocorticoid-induced osteoporosis by LMWHC-Pre upon 20 days of treatment. From this study, LMWHC-Pre may be employed as an effective potential drug candidate for the treatment of chronic renal disease.
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[Anti-tumor effect of cisplatin combined with DC vaccine on tumor-bearing mice].
Zhonghua Zhong Liu Za Zhi
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To explore the anti-tumor mechanism of the combination of cisplatin with DC vaccine in tumor-bearing mice.
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Association between the ERCC5 Asp1104His polymorphism and cancer risk: a meta-analysis.
PLoS ONE
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Excision repair cross complementing group 5 (ERCC5 or XPG) plays an important role in regulating DNA excision repair, removal of bulky lesions caused by environmental chemicals or UV light. Mutations in this gene cause a rare autosomal recessive syndrome, and its functional single nucleotide polymorphisms (SNPs) may alter DNA repair capacity phenotype and cancer risk. However, a series of epidemiological studies on the association between the ERCC5 Asp1104His polymorphism (rs17655, G>C) and cancer susceptibility generated conflicting results.
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[Therapeutic patient education in health centers in the Lorraine region of France - an EPATEL study].
Sante Publique
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The purpose of this paper is to map out the Therapeutic Patient Education activities of health centers in the Lorraine region of France in the months leading up to the promulgation of the HPST law (reform of the French Public Hospital system) defining and regulating TPE. A self-administered survey and interviews were conducted to identify the educational activities offered by the program (structure, training of professionals, links with general practitioners, etc.) and to quantify them (number of patients, classes, etc.). All health centers in Lorraine were contacted (193 centers in total). 93 centers responded and 48?reported TPE activities, with 131 functional TPE courses (or structured activities) and 40 projects. An analysis based on 8 quality criteria was carried out. The study found that 49 centers met at least 6 quality criteria and were close to the required quality standards. The study also found that the geographical distribution of TPE provision is unbalanced. The number of TPE beneficiaries in a given semester varies between 0 and 24 patients per 1,000 inhabitants (according to the health area), which amounts to 5% of chronically ill people in the Lorraine region.
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Tumor necrosis factor-? induced protein 8 polymorphism and risk of non-Hodgkins lymphoma in a Chinese population: a case-control study.
PLoS ONE
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Non-Hodgkins lymphoma (NHL) has been reported to be associated with autoimmune and pro-inflammatory response, and genetic polymorphisms of candidate genes involved in autoimmune and pro-inflammatory response may influence the susceptibility to NHL. To evaluate the role of such genetic variations in risk of NHL, we conducted a case-control study of 514 NHL patients and 557 cancer-free controls in a Chinese population.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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