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Find video protocols related to scientific articles indexed in Pubmed.
The small molecule probe PT-Yellow labels the renal proximal tubules in zebrafish.
Chem. Commun. (Camb.)
PUBLISHED: 11-20-2014
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We report the development of a small fluorescent molecule, BDNCA3-D2, herein referred to as PT-Yellow. Soaking zebrafish embryos in PT-Yellow or intraperitoneal injection into adults results in non-toxic in vivo fluorescent labeling of the renal proximal tubules, the major site of blood filtrate reabsorption and a common target of injury in acute kidney injury. We demonstrate the applicability of this new compound as a rapid and simple readout for zebrafish kidney filtration and proximal tubule reabsorption function.
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The complete mitochondrial genome of the Epinephelus akaara (Perciformes: Serranidae).
Mitochondrial DNA
PUBLISHED: 11-19-2014
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Abstract The complete mitochondrial genome of the Epinephelus akaara was presented in this study. The mitochondrial genome is 16,743?bp long and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. The gene order and composition of Epinephelus akaara mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 27.31% of A, 16.20% of C, 28.68% of T and 27.81% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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Influence of Recombinant Lentiviral Vector Encoding miR-15a/16-1 in Biological Features of Human Nasopharyngeal Carcinoma CNE-2Z Cells.
Cancer Biother. Radiopharm.
PUBLISHED: 11-19-2014
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Abstract MicroRNAs can function as tumor suppressor miRNAs. Bcl-2 is an antiapoptotic gene overexpressed in many tumors, including nasopharyngeal carcinoma (NPC). It is reported that microRNA-15a (miR-15a) and microRNA-16-1 (miR-16-1) could act as bcl-2 inhibitors. To investigate their effects on NPC, the authors used recombinant lentiviral vector to upregulate the expression of miR-15a/16-1 in NPC CNE-2Z cells. The authors divided cells into the control group, transfection group, radiotherapy group, and transfection-radiotherapy group. In this experiment, reverse transcription-quantitative polymerase chain reaction was used to detect the expression of miR-15a/16-1 and bcl-2 mRNA. Cell proliferation was analyzed by MTT assay. Flow cytometry was used to measure cell apoptosis. Radiosensitivity was measured using colony-forming experiment. The protein expression of bcl-2 was measured by western blot, the activation levels of caspase were detected by a spectrophotometric method. After transfection, cell proliferation was inhibited, while the apoptosis rate and radiosensitivity were increased. In addition, the activation of caspase-2 and caspase-3 was aggrandized correspondingly. Although the expression levels of bcl-2 mRNA in each group had no difference, the protein expression of bcl-2 was downregulated. These results suggested that miR-15a/16-1 could inhibit cell proliferation and increase the apoptosis and radiosensitivity of CNE-2 cells, by regulating the bcl-2 gene at post-transcriptional level and by increasing the activation of caspase-2 and caspase-3.
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Transmission of a Gaussian beam through a gyrotropic cylinder.
J Opt Soc Am A Opt Image Sci Vis
PUBLISHED: 11-18-2014
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An exact analytical solution to the transmission of a Gaussian beam through a gyrotropic cylinder is formulated in terms of a cylindrical vector wave function expansion form. By applying the continuous boundary conditions of electromagnetic fields, the unknown expansion coefficients of the scattered and internal fields are determined. For a localized beam model, numerical results are presented for the normalized near-surface and internal field intensity distributions, and the propagation characteristics are discussed concisely.
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Scattering of on-axis Gaussian beam by a uniaxial anisotropic object.
J Opt Soc Am A Opt Image Sci Vis
PUBLISHED: 11-18-2014
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Based on the extended boundary condition method, a semi-analytical solution to the scattering of an on-axis Gaussian beam by an arbitrarily shaped uniaxial anisotropic object is constructed. For the on-axis incident Gaussian beam, scattered fields as well as internal fields are expanded in terms of appropriate spherical vector wave functions, and the unknown expansion coefficients of the scattered fields are determined by using Schelkunoff's equivalence theorem and continuous boundary conditions. Numerical results of the normalized differential scattering cross section are presented, and the scattering characteristics are discussed concisely.
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miR-103 promotes 3T3-L1 cell adipogenesis through AKT/mTOR signal pathway with its target being MEF2D.
Biol. Chem.
PUBLISHED: 11-10-2014
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Abstract MicroRNAs are small non-coding RNAs that partially bind to the 3' untranslated (3'UTR) regions of target genes in animals and regulate protein production of the target transcripts. MiR-103 has been confirmed to play a critical role in lipid metabolism, however, the target genes and signaling pathway regulated by miR-103 is still unclear. In our experiment, we observed a positive function of miR-103 on the adipogenic differentiation of 3T3-L1 pre-adipocyte. Furthermore, we proved that this function of miR-103 was functioned through activating AKT/mTOR signal pathway and impairing target gene MEF2D. By inhibiting and over-expressing MEF2D gene, we found MEF2D had a negative role in regulating adipocyte key genes, and this function of MEF2D could be impaired by miR-103. In conclusion, we found miR-103 can promote 3T3-L1 cells differentiation by targeting MEF2D and activating AKT/mTOR signal pathway. These results will shed a light on further study of microRNAs.
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Profiling of stable isotope enrichment in specialized metabolites using liquid chromatography and multiplexed nonselective collision-induced dissociation.
Anal. Chem.
PUBLISHED: 10-23-2014
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Engineering of specialized metabolites in plants, microbes, and other organisms is hindered by significant knowledge gaps about metabolic pathways responsible for metabolite accumulation and degradation. While isotopic tracers have provided important information about metabolic fluxes in central metabolism, limitations of mass spectrometric strategies for quantifying stable isotope incorporation into both intact metabolites and specific substructures have slowed extension of these techniques to large specialized metabolites. This report describes the application of electrospray ionization with data-independent multiplexed nonselective collision induced dissociation (CID) on a time-of-flight mass spectrometer. This strategy yields quasi-simultaneous collection, on the chromatographic time scale, of mass spectra with different degrees of fragment ion formation without biases introduced by precursor mass selection or selective ion activation and provides measurements of stable isotope enrichments in intact metabolites and individual substructures. The utility and precision of these analyses is demonstrated by labeling acylsugar metabolites in glandular trichomes of tomato (Solanum lycopersicum) using (13)CO2 and analyzing (13)C enrichments in acylsugar specialized metabolites. The high precision and avoidance of mass bias provide a promising tool for extending metabolic flux analyses to complex specialized metabolites in a wide range of organisms.
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Nickel exposure is associated with the prevalence of type 2 diabetes in Chinese adults.
Int J Epidemiol
PUBLISHED: 10-18-2014
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Nickel exposure can induce hyperglycaemia in rodents, but little is known about its association with abnormal glucose metabolism in humans. We aimed to investigate the association of nickel exposure with the prevalence of type 2 diabetes in Chinese adults.
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A computational frame and resource for understanding the lncRNA-environmental factor associations and prediction of environmental factors implicated in diseases.
Mol Biosyst
PUBLISHED: 10-14-2014
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The complex traits of an organism are associated with a complex interplay between genetic factors (GFs) and environmental factors (EFs). However, compared with protein-coding genes and microRNAs, there is a paucity of computational methods and bioinformatic resource platform for understanding the associations between lncRNA and EF. In this study, we developed a novel computational method to identify potential associations between lncRNA and EF, and released LncEnvironmentDB, a user-friendly web-based database aiming to provide a comprehensive resource platform for lncRNA and EF. Topological analysis of EF-related networks revealed the small world, scale-free and modularity structure. We also found that lncRNA and EF significantly enriched interacting miRNAs are functionally more related by analyzing their related diseases, implying that the predicted lncRNA signature of EF can reflect the functional characteristics to some degree. Finally, we developed a random walk with a restart-based computational model (RWREFD) to predict potential disease-related EFs by integrating lncRNA-EF associations and EF-disease associations. The performance of RWREFD was evaluated by experimentally verified EF-disease associations based on leave-one-out cross-validation and achieved an AUC value of 0.71, which is higher than randomization test, indicating that the RWREFD method has a reliable and high accuracy of prediction. To the best of our knowledge, LncEnvironmentDB is the first attempt to predict and house the experimental and predicted associations between lncRNA and EF. LncEnvironmentDB is freely available on the web at .
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The complete mitochondrial genome of the Epinephelus lanceolatus (Perciformes: Serranidae).
Mitochondrial DNA
PUBLISHED: 10-07-2014
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Abstract The complete mitochondrial genome of the Epinephelus lanceolatus was presented in this study. The mitochondrial genome is 16,743?bp long and consists of 13 protein-coding genes, two rRNA genes, 22 tRNA genes and a control region. The gene order and composition of E. lanceolatus mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 26.55% of A, 15.02% of C, 29.67% of T and 28.76% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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[Two types of digestive tract reconstruction after proximal gastrectomy for early gastroesophageal junction adenocarcinoma: a randomized controlled study].
Zhonghua Wei Chang Wai Ke Za Zhi
PUBLISHED: 10-03-2014
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To investigate the better method of digestive tract reconstruction in proximal gastrectomy for early gastroesophageal junction adenocarcinoma.
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Hypoglycemic, hypolipidemic and antioxidant effects of Sarcandra glabra polysaccharide in type 2 diabetic mice.
Food Funct
PUBLISHED: 09-26-2014
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Sarcandra glabra (Thunb.) Nakai is a traditional Chinese herbal medicine and dietary supplement used for treating several diseases. The anti-diabetic activity of S. glabra polysaccharides is reported for the first time. The in vitro ?-glucosidase inhibition assay indicated that the acidic S. glabra polysaccharide (SGP-2) has an IC50 of 87.06 ± 11.76 ?g mL(-1), which was much lower than acarbose at 338.90 ± 46.86 ?g mL(-1). Moreover, high fat diet (HFD) with streptozotocin (STZ) induced diabetic mice were administered SGP-2 (150, 300, or 600 mg kg(-1) per day, respectively) for 3 weeks. Postprandial blood glucose levels (PBGL), total cholesterol, triglyceride and free fatty acid levels in diabetic mice treated with SGP-2 were significantly decreased (p < 0.05) compared to those of the model group. The results of the oral glucose tolerance test (OGTT) and the homeostasis model assessment-insulin resistance (HOMA-IR) index indicated that SGP-2 could significantly improve (p < 0.05) the insulin resistance and glucose tolerance in diabetic mice. Furthermore, the activities of antioxidant enzymes, hexokinase and pyruvate kinase were significantly increased (p < 0.05) in SGP-2 treated groups. Thus we proposed that SGP-2 exerted hypoglycemic activity by relieving insulin resistance, reducing postprandial blood glucose levels and ameliorating lipid metabolism, as well as alleviating oxidative stress. These data suggested that SGP-2 with anti-hyperglycemic activity could be used in medicinal preparations for diabetes mellitus and its complications.
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The complete mitochondrial genome of the Drepane punctata (Perciformes: Drepanidae).
Mitochondrial DNA
PUBLISHED: 09-16-2014
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Abstract The complete mitochondrial genome of the Drepane punctata was presented in our study. The mitochondrial genome is 16,397?bp long and consists of 13 protein-coding genes, two rRNA genes, 16 tRNA genes and a control region. The gene order and composition of D. punctata mitochondrial genome was different from that of most other vertebrates. The nucleotide compositions of the light strand are 24.56% of A, 16.02% of C, 27.81% of T and 31.61% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and five tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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The complete mitochondrial genome of the orange-spotted grouper Epinephelus coioides (Perciformes, Serranidae).
Mitochondrial DNA
PUBLISHED: 09-12-2014
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Abstract We present the complete mitochondrial genome of Epinephelus coioides in this study. The mitochondrial genome is 16,458?bp in length, containing 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. All of them are encoded on the heavy strand except ND6 and 8 tRNA genes on the light strand. The nucleotide compositions of the light strand are 28.88% of G, 28.75% of T, 26.58% of A and 15.80% of C. All the protein-coding genes share ATG initiation codon except ATP6 with CTG and COXI with GTG, and four types of inferred termination codons are T (ND2, COXII, ND3, ND4 and Cytb), TA (COXIII), TAA (ND1, ATP8, ND4L, ND5, ATP6 and COXI) and TAG (ND6). There are 10 intergenic spacers, 5 gene overlaps and no tandem repeat sequence. Gene arrangement and distribution are consistent with the typical vertebrates.
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The complete mitochondrial genome of the Rhabdosargus sarba (Perciformes: Sparidae).
Mitochondrial DNA
PUBLISHED: 09-11-2014
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Abstract The complete mitochondrial genome of the Rhabdosargus sarba was presented in our study. The mitochondrial genome is 16,644?bp long and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. The gene order and composition of R. sarba mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 27.01% of A, 17.96% of C, 26.02% of T and 29.01% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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miR-125b inhibits hepatitis B virus expression in vitro through targeting of the SCNN1A gene.
Arch. Virol.
PUBLISHED: 08-31-2014
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microRNAs (miRNAs) are small noncoding RNAs that modulate gene expression at the posttranscriptional level, playing an important role in many diseases. However, reports concerning the role of miRNA in hepatitis B virus (HBV) infection are limited. miRNA chips were used to investigate miRNA changes during HBV infection in vitro. Bioinformatics analysis was used to explore possible miRNA and target genes during HBV infection. The expression of miR-125b and its potential target gene, sodium channel, non-voltage-gated 1 alpha (SCNN1A), was further analyzed. A total of 136 miRNAs were analyzed in an HBV transient transfection model (HepG2-HBV1.3), and 78 miRNAs were differentially expressed in HepG2.2.15 cells compared with HepG2 cells. miR-125b expression was decreased in both HepG2-HBV1.3 and HepG2.2.15 cells, and ectopic expression of miR-125b inhibited HBV DNA intermediates and secretion of HBsAg and HBeAg. miR-125b also inhibited the mRNA and protein levels of SCNN1A. Using a dual luciferase reporter system, we found that SCNN1A was one of the targets of miR-125b. In this study, we found that miR-125b inhibits HBV expression in vitro by regulating SCNN1A expression.
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A multi-stimuli responsive gold nanocage-hyaluronic platform for targeted photothermal and chemotherapy.
Biomaterials
PUBLISHED: 08-28-2014
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Noninvasive and pinpointed intracellular drug release that responds to multiple stimulus is still a formidable challenge for cancer therapy. Herein, we reported a multi-stimuli responsive platform based on drug loaded gold nanocages @ hyaluronic acid (AuNCs-HA) for pinpointed intracellular drug release. These well-prepared nanohybrids could specifically recognize cancer cells via HA-CD44 interactions and be efficiently endocytosed by receptor-mediated process. Subsequently, the coated HA molecules could be degraded in lysosomes, resulting in the release of encapsulated drug. In addition, by taking advantage of the excellent photothermal properties, the AuNCs could accelerate the release of encapsulated drug and induce a higher therapeutic efficacy upon near-infrared (NIR) irradiation. In vitro results confirmed that the encapsulated drug could only be pinpointedly released in intracellular environments, which permitted high therapeutic efficacy against cancer cells and minimized the side effects. Importantly, as compared to that of the two therapies independently, a complete inhibition of tumor growth treated with the combination of chemotherapy and photothermal therapy was observed in vivo. Taken together, our present study provides new insights into developing pinpointed, multi-stimuli responsive intracellular drug release systems for synergistic cancer therapy.
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Analysis of POFUT1 gene mutation in a Chinese family with Dowling-Degos disease.
PLoS ONE
PUBLISHED: 08-26-2014
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Dowling-Degos disease (DDD) is an autosomal dominant genodermatosis characterized by reticular pigmented anomaly mainly affecting flexures. Though KRT5 has been identified to be the causal gene of DDD, the heterogeneity of this disease was displayed: for example, POFUT1 and POGLUT1 were recently identified and confirmed to be additional pathogenic genes of DDD. To identify other DDD causative genes, we performed genome-wide linkage and exome sequencing analyses in a multiplex Chinese DDD family, in which the KRT5 mutation was absent. Only a novel 1-bp deletion (c.246+5delG) in POFUT1 was found. No other novel mutation or this deletion was detected in POFUT1 in a second DDD family and a sporadic DDD case by Sanger Sequencing. The result shows the genetic-heterogeneity and complexity of DDD and will contribute to the further understanding of DDD genotype/phenotype correlations and to the pathogenesis of this disease.
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Effect of Tea Polyphenols on Lipid Peroxidation and Antioxidant Activity of Litchi (Litchi chinensis Sonn.) Fruit during Cold Storage.
Molecules
PUBLISHED: 08-23-2014
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To understand the potential of application of tea polyphenols to the shelf life extension and quality maintenance of litchi (Litchi chinensis Sonn.) fruit, the fruits were dipped into a solution of 1% tea phenols for 5 min before cold storage at 4 °C. Changes in browning index, contents of anthocyanins and phenolic compounds, superoxide dismutase (SOD) and peroxidase (POD) activities, O2.- production rate and H2O2 content, levels of relative leakage rate and lipid peroxidation, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were measured after 0, 10, 20 and 30 days of cold storage. The results showed that application of tea polyphenols markedly delayed pericarp browning, alleviated the decreases in contents of total soluble solids (TSS) and ascorbic acid, and maintained relatively high levels of total phenolics and anthocyanins of litchi fruit after 30 days of cold storage. Meanwhile, the treatment reduced the increases in relative leakage rate and lipid peroxidation content, delayed the increases in both O2.- production rate and H2O2 contents, and increased SOD activity but reduced POD activity throughout this storage period. These data indicated that the delayed pericarp browning of litchi fruit by the treatment with tea polyphenols could be due to enhanced antioxidant capability, reduced accumulations of reactive oxygen species and lipid peroxidation, and improved membrane integrity.
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Taxonomy and phylogeny of two species of the genus Deviata (Protista, Ciliophora) from China, with description of a new soil form, Deviata parabacilliformis sp. nov.
Int. J. Syst. Evol. Microbiol.
PUBLISHED: 08-19-2014
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The morphology and morphogenesis of a soil hypotrichous ciliate, Deviata parabacilliformis sp. nov., isolated from northern China, were investigated. D. parabacilliformis measures about 75-210×25-60 µm in vivo, with an elongate and flexible body. It possesses one right marginal row, two to four left marginal rows and three dorsal kineties. The main morphogenetic features of D. parabacilliformis are: (i) the oral primordium originates de novo; (ii) anlage IV of the opisthe originates from parental frontoventral row V, anlage V originates de novo, and anlage VI forms from frontoventral row VI; and (iii) anlage I of the proter originates from the anterior portion of the parental paroral, anlage II originates from the buccal cirrus, anlage III originates from the parabuccal cirri, anlage IV originates from parental frontoventral row IV and anlage V forms from the anterior of parental frontoventral row VI. The morphology of an edaphic population of another species of the genus Deviata, Deviata bacilliformis (Gelei 1954) Eigner 1995, was also investigated. This work also provides the first record of SSU rRNA gene sequences for species of the genus Deviata. Molecular phylogenetic analysis suggests that Deviata is not monophyletic, and its position is poorly resolved due to weak phylogenetic signal of the 18S marker in the Stichotrichida.
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Evolution of the Germline Actin Gene in Hypotrichous Ciliates: Multiple Nonscrambled IESs at Extremely Conserved Locations in Two Urostylids.
J. Eukaryot. Microbiol.
PUBLISHED: 08-08-2014
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In hypotrichous ciliates, macronuclear chromosomes are gene-sized, and micronuclear genes contain short, noncoding internal eliminated segments (IESs) as well as macronuclear-destined segments (MDSs). In the present study, we characterized the complete macronuclear gene and two to three types of micronuclear actin genes of two urostylid species, i.e. Pseudokeronopsis rubra and Uroleptopsis citrina. Our results show that (1) the gain/loss of IES happens frequently in the subclass Hypotrichia (formerly Stichotrichia), and high fragmentation of germline genes does not imply for gene scrambling; and (2) the micronuclear actin gene is scrambled in the order Sporadotrichida but nonscrambled in the orders Urostylida and Stichotrichida, indicating the independent evolution of MIC-actin gene patterns in different orders of hypotrichs; (3) locations of MDS-IES junctions of micronuclear actin gene in coding regions are conserved among closely related species.
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Perinatal lead (Pb) exposure results in sex-specific effects on food intake, fat, weight, and insulin response across the murine life-course.
PLoS ONE
PUBLISHED: 08-08-2014
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Developmental lead (Pb) exposure has been associated with lower body weight in human infants and late onset obesity in mice. We determined the association of perinatal Pb exposure in mice with changes in obesity-related phenotypes into adulthood. Mice underwent exposure via maternal drinking water supplemented with 0 (control), 2.1 (low), 16 (medium), or 32 (high) ppm Pb-acetate two weeks prior to mating through lactation. Offspring were phenotyped at ages 3, 6, and 9 months for energy expenditure, spontaneous activity, food intake, body weight, body composition, and at age 10 months for glucose tolerance. Data analyses were stratified by sex and adjusted for litter effects. Exposed females and males exhibited increased energy expenditure as compared to controls (p<0.0001 for both). In females, horizontal activity differed significantly from controls (p?=?0.02) over the life-course. Overall, food intake increased in exposed females and males (p<0.0008 and p<0.0001, respectively) with significant linear trends at 9 months in females (p?=?0.01) and 6 months in males (p<0.01). Body weight was significantly increased in males at the medium and high exposures (p?=?0.001 and p?=?0.006). Total body fat differed among exposed females and males (p<0.0001 and p<0.0001, respectively). Insulin response was significantly increased in medium exposure males (p<0.05). Perinatal Pb exposure at blood lead levels between 4.1 µg/dL and 32 µg/dL is associated with increased food intake, body weight, total body fat, energy expenditure, activity, and insulin response in mice. Physiological effects of developmental Pb exposure persist and vary according to sex and age.
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A bioinformatics method for predicting long noncoding RNAs associated with vascular disease.
Sci China Life Sci
PUBLISHED: 08-08-2014
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Long noncoding RNAs (lncRNAs) play important roles in human diseases including vascular disease. Given the large number of lncRNAs, however, whether the majority of them are associated with vascular disease remains unknown. For this purpose, here we present a genomic location based bioinformatics method to predict the lncRNAs associated with vascular disease. We applied the presented method to globally screen the human lncRNAs potentially involved in vascular disease. As a result, we predicted 3043 putative vascular disease associated lncRNAs. To test the accuracy of the method, we selected 10 lncRNAs predicted to be implicated in proliferation and migration of vascular smooth muscle cells (VSMCs) for further experimental validation. The results confirmed that eight of the 10 lncRNAs (80%) are validated. This result suggests that the presented method has a reliable prediction performance. Finally, the presented bioinformatics method and the predicted vascular disease associated lncRNAs together may provide helps for not only better understanding of the roles of lncRNAs in vascular disease but also the identification of novel molecules for the diagnosis and therapy of vascular disease.
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The complete mitochondrial genome of the Piaractus brachypomus (Characiformes: Characidae).
Mitochondrial DNA
PUBLISHED: 08-05-2014
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Abstract The complete mitochondrial genome of the Piaractus brachypomus is described in the present study. The mitochondrial genome is 16,561?bp long and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. The P. brachypomus mitochondrial genome shows the similar gene order and composition with those of most other vertebrates. The nucleotide compositions of the light strand in descending order is 31.57% of A, 26.19% of C, 26.18% of T and 16.06% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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A Molecular Phylogenetic Investigation of Bakuella, Anteholosticha, and Caudiholosticha (Protista, Ciliophora, Hypotrichia) Based on Three Gene Sequences.
J. Eukaryot. Microbiol.
PUBLISHED: 07-30-2014
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Traditionally classifications of the Urostyloida have been mainly based on morphology and morphogenesis. Recent molecular phylogenetic analyses have been largely based on single-gene data for a limited number of taxa. Consequently, incongruence has arisen between the morphological/morphogenetic and the molecular data. In this study, the three phylogenetic markers (SSU rDNA, ITS1-5.8S-ITS2 region and LSU-rDNA) of three urostyloid genera represented by four species (Bakuella granulifera, Anteholosticha monilata, Caudiholosticha sylvatica and C. tetracirra) were sequenced in order to investigate their phylogeny. The results show that: (1) all three genera should be regarded as the members of the order Urostyloida within the subclass Hypotrichia, as indicated by morphological characters; (2) phylogenetic analyses and sequence similarities both indicate that neither Anteholosticha nor Caudiholosticha are monophyletic and the systematic assignment of both genera awaits further evaluation; and (3) Bakuella has a closer relationship with Urostyla than with bakuellids (e.g. Apobakuella and Metaurostylopsis), suggesting Bakuella may belong to the family Urostylidae rather than the family Bakuellidae. This article is protected by copyright. All rights reserved.
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Multifunctional upconverting nanoparticles for near-infrared triggered and synergistic antibacterial resistance therapy.
Chem. Commun. (Camb.)
PUBLISHED: 07-29-2014
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To integrate photodynamic therapy with photothermal therapy for improved multidrug-resistant bacteria therapy, we have constructed a novel multifunctional core/satellite nanostructure by decorating CuS nanoparticles onto the surface of NaYF4:Mn/Yb/Er@photosensitizer doped SiO2. This system exhibited a superior antibacterial activity towards drug-resistant Staphylococcus aureus and Escherichia coli.
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Sandwich-type Au-PEI/DNA/PEI-Dexa nanocomplex for nucleus-targeted gene delivery in vitro and in vivo.
ACS Appl Mater Interfaces
PUBLISHED: 07-24-2014
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Many synthetic Au-based cationic nanoparticles (AuNPs) for nonviral gene delivery show high efficiency in vitro, but their excessive charge density, harsh reducing conditions, and nontarget delivery prevent their application in vivo. Herein, we constructed a sandwich-type layered polyethylenimine (PEI)-coated gold nanocomposite outerlaid with a nucleus-targeted Dexamethasone (Dexa), namely, Au-PEI/DNA/PEI-Dexa nanocomplex, for DNA delivery system using a low molecular weight PEI as a mild reducing agent. The nucleus-targeting Au-PEI/DNA/PEI-Dexa nanocomplex with low positive charge and low cytotoxicity condensed DNA and protected from enzymatic degradation. In vitro transfection studies demonstrated that Au-PEI/DNA/PEI-Dexa nanocomplex exhibited much more efficient nucleus transfection than Au-PEI/DNA/PEI without nucleus-targeted residues and commercially available PEI 25 kDa due to the Dexa targeting of the nucleus. Furthermore, the nanocomplex markedly transfected pTRAIL (TRAIL = tumor-necrosis-factor-related apoptosis-inducing ligand) to tumors in vivo and subsequently inhibited the tumor growth with minimal side effects. These findings suggest that nucleus-targeting Au-PEI/DNA/PEI-Dexa ternary complexes have promising potential in gene delivery.
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The complete mitochondrial genome of the Siganus canaliculatus (Perciformes: Siganidae).
Mitochondrial DNA
PUBLISHED: 07-02-2014
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Abstract The complete mitochondrial genome of the Siganus canaliculatus was presented in this study. The mitochondrial genome is 16,492?bp long and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. The gene order and composition of Siganus canaliculatus mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand in descending order is 29.74% of C, 28.15% of A, 25.43% of T and 16.68% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and 8 tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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Generation of haploid spermatids with fertilization and development capacity from human spermatogonial stem cells of cryptorchid patients.
Stem Cell Reports
PUBLISHED: 06-27-2014
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Generation of functional spermatids from azoospermia patients is of unusual significance in the treatment of male infertility. Here, we report an efficient approach to obtain human functional spermatids from cryptorchid patients. Spermatogonia remained whereas meiotic germ cells were rare in cryptorchid patients. Expression of numerous markers for meiotic and postmeiotic male germ cells was enhanced in human spermatogonial stem cells (SSCs) of cryptorchidism patients by retinoic acid (RA) and stem cell factor (SCF) treatment. Meiotic spreads and DNA content assays revealed that RA and SCF induced a remarkable increase of SCP3-, MLH1-, and CREST-positive cells and haploid cells. Single-cell RNA sequencing analysis reflected distinct global gene profiles in embryos derived from round spermatids and nuclei of somatic cells. Significantly, haploid spermatids generated from human SSCs of cryptorchid patients possessed fertilization and development capacity. This study thus provides an invaluable source of autologous male gametes for treating male infertility in azoospermia patients.
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The complete mitochondrial genome of the Pampus nozawae (Perciformes: Stromateidae).
Mitochondrial DNA
PUBLISHED: 06-25-2014
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Abstract The complete mitochondrial genome of the Pampus nozawae was presented in this study. The mitochondrial genome is 16,556?bp long and consists of 13 protein-coding genes, two rRNA genes, 22 tRNA genes and a control region. The gene order and composition of Pampus nozawae mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 30.01% of A, 27.51% of C, 27.29% of T and 15.18% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and seven tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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Subcutaneous fat necrosis of the newborn associated with hypercalcemia after therapeutic hypothermia.
J La State Med Soc
PUBLISHED: 06-10-2014
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Subcutaneous fat necrosis of the newborn (SCFN) is a rare, benign, and self-limiting panniculitis of neonates that presents in the first few weeks of life and is most commonly associated with birth asphyxia and meconium aspiration. There have been few case reports of SCFN following therapeutic hypothermia. With the increasing use of therapeutic whole-body hypothermia, SCFN may become more prevalent. The differential diagnosis of SCFN can be broad, and clinicopathologic correlation is essential to make the correct and timely diagnosis. Clinicians should be aware of this rare disease and its potential serious complication hypercalcemia.
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Phylogenetic and genetic characterization of Acidithiobacillus strains isolated from different environments.
World J. Microbiol. Biotechnol.
PUBLISHED: 05-30-2014
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To study the phylogenetic relationships and genetic heterogeneity of 21 Acidithiobacillus strains isolated from different environments, we amplified and sequenced the 16S-23S rRNA gene intergenic spacers (ITS) of all these strains. These sequence data, combined with related sequences available from GenBank, were divided into six phylogenetic groups by 16S rRNA gene and by 16S-23S rRNA gene sequence analysis. The results of phylogenetic analysis were consistent with those obtained by repetitive element PCR and arbitrarily primed PCR. In this research, the Acidithiobacillus ferrooxidans (A. ferrooxidans) strains were always separated into two groups in phylogenetic and cluster analyses. Genotypic analyses of the genes rusA, rusB, hip and iro suggest that these two groups may have different biochemical mechanisms for oxidizing ferrous iron. Strains in one A. ferrooxidans group were detected with rusA gene that encodes rusticyanin A which plays a very important role in the iron respiratory chain. The second A. ferrooxidans group was found to contain rusB gene which encode a homologous protein (RusB). The data suggested that ITS-based phylogeny is an effective tool to elucidate the relationships of Acidithiobacillus and that a different iron oxidation pathway may exist in different A. ferrooxidans groups.
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[Promotion of postoperative recovery with fast track surgery for gastric cancer patients undergoing gastrectomy: a prospective randomized controlled study].
Zhonghua Wei Chang Wai Ke Za Zhi
PUBLISHED: 05-27-2014
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To study the safety and feasibility of fast track surgery (FTS) in the promotion of postoperative recovery for gastric cancer patients undergoing gastrectomy.
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Optimal packing of a rotating packed bed for H(2)S removal.
Environ. Sci. Technol.
PUBLISHED: 05-27-2014
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The existence of H2S in a system could lead to catalyst deactivation, pipeline corrosion, and environmental pollution. A rotating packed bed (RPB), a novel reactor with high mass transfer efficiency and small dimension, is employed in this study to remove H2S. For RPB, the most significant section for mass transfer is the end-effect zone of packing. A mathematical model for liquid flow in the packing is established to quantify the length of the end-effect zone. A simple and effective visual experimental method is then proposed to investigate the end-effect zone in the RPB. A gas-liquid mass transfer experiment is finally employed to confirm the validity of the proposed mathematical model. With the aid of this model, the length of packing of a RPB used for pilot-scale H2S removal is optimized. The optimized RPB removes 99.8% of H2S (15 vol % to 0.03 vol %) from the system. The proposed model can help optimize the design of a RPB reactor.
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Inferring novel lncRNA-disease associations based on a random walk model of a lncRNA functional similarity network.
Mol Biosyst
PUBLISHED: 05-23-2014
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Accumulating evidence demonstrates that long non-coding RNAs (lncRNAs) play important roles in the development and progression of complex human diseases, and predicting novel human lncRNA-disease associations is a challenging and urgently needed task, especially at a time when increasing amounts of lncRNA-related biological data are available. In this study, we proposed a global network-based computational framework, RWRlncD, to infer potential human lncRNA-disease associations by implementing the random walk with restart method on a lncRNA functional similarity network. The performance of RWRlncD was evaluated by experimentally verified lncRNA-disease associations, based on leave-one-out cross-validation. We achieved an area under the ROC curve of 0.822, demonstrating the excellent performance of RWRlncD. Significantly, the performance of RWRlncD is robust to different parameter selections. Predictively highly-ranked lncRNA-disease associations in case studies of prostate cancer and Alzheimer's disease were manually confirmed by literature mining, providing evidence of the good performance and potential value of the RWRlncD method in predicting lncRNA-disease associations.
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The complete mitochondrial genome of the Platax teira (Osteichthyes: Ephippidae).
Mitochondrial DNA
PUBLISHED: 05-22-2014
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Abstract We presented the complete mitochondrial genome of the Platax teira in this study. The mitochondrial genome is 16,561?bp long and consists of 13 protein-coding genes, two rRNA genes, 22 tRNA genes and a control region. The gene order and composition of Epinephelus tukula mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 28.80% of A, 31.16% of C, 24.44% of T and 15.60% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and seven tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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Enhanced shRNA Delivery and ABCG2 Silencing by Charge-Reversible Layered Nanocarriers.
Small
PUBLISHED: 05-20-2014
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Polycationic vectors have been used to deliver short hairpin RNAs (shRNAs) to knock-down genes for cancer therapies, but their inefficiency in lysosomal escape and shRNA release causes their low gene transcription efficiency. Herein, a three-layered polyethyleneimine (PEI)-coated gold nanocomplex interlaid with a pH-responsive charge-reversible chitosan-aconitic anhydride (CS-Aco) is constructed: a Au-PEI/CS-Aco/PEI/shRNA nanoparticle. The negatively charged CS-Aco hydrolyzes into positively charged CS in lysosomes, causing the nanocomposite to disassemble. The released Au-PEI nanoparticles efficiently rupture the lysosomes and thus release the PEI/shRNA polyplexes into cytoplasm, where they quickly disassociate because the PEI chains are short (1.2 kDa). As a consequence, the nanocomplexes display higher shRNA delivery efficiency than the 25 kDa PEI, and efficiently deliver shABCG2 to tumors and markedly silence ABCG2 expression, which sensitizes HepG2 cells to the drugs with minimal toxicity.
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The complete mitochondrial genome of the Hemibagrus guttatus (Teleostei, Bagridae).
Mitochondrial DNA
PUBLISHED: 05-10-2014
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Abstract The complete mitochondrial genome of the Hemibagrus guttatus was presented in this study. The mitochondrial genome is 16,523?bp long and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. The gene order and composition of H. guttatus mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 31.73% of A, 25.42% of C, 28.02% of T and 14.83% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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The complete mitochondrial genome of the Culter recurviceps (Teleostei, Cyprinidae).
Mitochondrial DNA
PUBLISHED: 05-10-2014
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Abstract We present the complete mitochondrial genome of the Culter recurviceps in this study. The mitochondrial genome is 16,622?bp long and consists of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. The gene order and composition of C. recurviceps mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 31.36% of A, 24.77% of T, 27.77% of C and 16.09% of G. With the exception of ND6 and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand. Two copies of tandem repeat sequence (24?bp) was observed in the end of the 123?bp in control region.
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Mutation analysis of the IL36RN gene in Chinese patients with generalized pustular psoriasis with/without psoriasis vulgaris.
J. Dermatol. Sci.
PUBLISHED: 04-30-2014
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Generalized pustular psoriasis (GPP) is a rare type of psoriasis with potentially life-threatening implications. Mutations in IL36RN gene have been suggested to be causative or predisposing factors for GPP.
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Synthesis and characterization of C70-corrole--a new electron transfer dyad.
J Nanosci Nanotechnol
PUBLISHED: 04-25-2014
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A new electron transfer dyad, covalently linked C70-corrole, was prepared via C70 and 10-(4-Formylaryl)-5,15-bis(pentafluorophenyl). The structures and the properties of the new material were investigated by HPLC, MALDI-TOF-MS, UV-Vis-NIR spectroscopy, NMR, fluorescence analysis and CV/DPV. The free-energy of C70-corrole calculated by employing the redox potentials and singlet excited-state energy suggested the possibility of electron transfer from the excited singlet state of corrole to the fullerene entity, which agreed with the results of the theoretical calculation.
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Ion Torrent next-generation sequencing reveals the complete mitochondrial genome of black and reddish morphs of the Coral Trout Plectropomus leopardus.
Mitochondrial DNA
PUBLISHED: 04-18-2014
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Abstract Using Ion Torrent next-generation sequencing (NGS) technology, we sequenced the complete mitochondrial genome (mitogenome) of black and reddish morphs of the coral trout Plectropomus leopardus. High-throughput sequencing generated a total of 958,614 sequence reads covering 164.80?Mb of two mitogenomes with a coverage of 4800X. Thirty-seven mitochondrial genes and gene order of P. leopardus was quite similar to that of other teleostean fishes. Most genes were either abutted or overlapped, and all the protein-coding genes began with an ATG start codon except for COX1 and ATP6. The number of stop codon was different for the black and reddish P. leopardus. Comparisons between the mitochondrial sequences of the two morphs revealed a total of 74 variable sites and one indel. Nucleotide diversity across protein-coding gene varied from 0.0006 (16s rRNA) to 0.0070 (Cytochrome b). As expected, the highest level of nucleotide diversity (0.0291) was detected in the control region. Our results demonstrate the NGS technology based on Ion torrent platform can be used to assemble the mitogenome of fish species.
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Self-assembly of silver nanoparticles illuminated by a visible-near-infrared lamp.
J Nanosci Nanotechnol
PUBLISHED: 04-17-2014
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Silver nanoparticles (NPs) with diameters of 5-10 nm were synthesized by using formaldehyde (HCHO) to reduce silver nitrate (AgNO3) and polyethylenemine (PEI) as a stabilizer. It was found that on the carbon-supported cupper grid (Cu/polyvinylformal (PVF)/C) and under a visible-near-infrared (VIS-NIR, 400-860 nm) lamp illuminating at about 75 degrees C for six minutes, the spherical Ag NPs self-assembled into bigger NPs, triangular nanoprisms, or structures like clock hands. On the glass slides, the NPs firstly self-assembled into spherical particles with 3-6 microm in sizes, then assembled into chain structures comprised of 3-20 spherical particles after being illuminated by the VIS-NIR lamp for about nine minutes. The violent thermal motion of the NPs after being illuminated by the VIS-NIR lamp leads to the self-assembly and the shapes of the self-assembled particles are related to the interactions between the substrates and the samples.
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The ERK/eIF4F/Bcl-XL pathway mediates SGP-2 induced osteosarcoma cells apoptosis in vitro and in vivo.
Cancer Lett.
PUBLISHED: 04-16-2014
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The aim of this study is to assess the molecular foundation of anti-tumor activity of SGP-2 in osteosarcoma cells. SGP-2 significantly blocks cell proliferation in human osteosarcoma U2OS cell model and inhibits tumor growth without causing apparent toxicity effect in mouse sarcoma S-180 cell-derived tumor model. Moreover, SGP-2 induces intrinsic apoptosis including the activation of caspase-3/7/9, the loss of mitochondrial transmembrane potential (??m), and the release of cytochrome c from mitochondrion, controlled by the down-regulation of B-cell lymphoma-extra large (Bcl-XL). Further research reveals that SGP-2 inhibits the assembly of eukaryotic initiation factor 4F (eIF4F) complex which is responsible for the decline of Bcl-XL. Finally, extracellular-signal-regulated kinase (ERK) controls SGP-2 induced intrinsic apoptosis. Taken together, SGP-2 exerts anti-tumor effect through intrinsic apoptotic pathway controlled by ERK/eIF4F/Bcl-XL pathway.
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Coupled selection of protein solubility in E. coli using uroporphyrinogen III methyltransferase as red fluorescent reporter.
J. Biotechnol.
PUBLISHED: 04-14-2014
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Uroporphyrinogen III methyltransferase (UMT) is a novel reporter owing to the catalytic products accumulated in cells emitting red florescence. Overexpression of UMT confers resistance of the Escherichia coli cells to potassium tellurite that inhibits cell growth. In this study, we applied UMT reporter for monitoring protein solubility of MBP or TEV protease variants under different expression conditions, as well as 12 maize proteins with either the designed linker or N-terminal SUMO tag. Effects of five enzymes involved in heme and siroheme biosynthesis on the reporter were also investigated. With increasing concentrations of potassium tellurite, colony numbers of the mixed cells expressing the selected five proteins with different solubility were decreased, but colonies displaying red fluorescence was identified to be produced the protein with relatively high solubility. The developed UMT reporter system is sensitive for monitoring protein solubility based on coupled fluorescence and chemical selection.
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The complete mitochondrial genome of the Epinephelus tukula (Perciformes, Serranidae).
Mitochondrial DNA
PUBLISHED: 04-09-2014
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Abstract The complete mitochondrial genome of the Epinephelus tukula was presented in this study. The mitochondrial genome is 16,503?bp long and consists of 13 protein-coding genes, two rRNA genes, 22 tRNA genes and a control region. The gene order and composition of Epinephelus tukula mitochondrial genome was similar to that of most other vertebrates. The nucleotide compositions of the light strand are 28.37% of A, 29.00% of C, 26.36% of T and 16.26% of G. With the exception of the NADH dehydrogenase subunit 6 (ND6) and seven tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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Morphology and Molecular Phylogeny of Three Cyrtophorid Ciliates (Protozoa, Ciliophora) from China, Including Two New Species, Chilodonella parauncinata sp. n. and Chlamydonella irregularis sp. n.
J. Eukaryot. Microbiol.
PUBLISHED: 04-05-2014
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This study investigated the morphology and molecular characteristics of three interesting free-living cyrtophorid ciliates, including two new species, isolated from China: Chilodonella parauncinata sp. n. can be identified by its elongated body shape, with a sharp protrusion in the left anterior part, cell size ca. 60 × 25 ?m in vivo, five right and 6-7 left kineties with kinetosomes densely arranged, and a curved cyrtos. Chlamydonella irregularis sp. n. differs from its congeners by the oval body shape, cell size 50-60 × 25-40 ?m in vivo, irregular shape of macronucleus, 30-40 club-shaped ventral protuberances, and 17 somatic kineties. Two isolates of Chlamydonella derouxi Song, 2003, collected from an intertidal area in Shandong and a mangrove wetland in Guangdong respectively, correspond well with two previous descriptions, but differ in comprising more basal bodies in left and right equatorial fragments and in having more finger-like protuberances on the ventral side. Phylogenetic analyses based on the small subunit rRNA gene sequences showed that C. parauncinata sp. n. clustered with Chilodonella uncinata, but was a well-outlined species of the genus, and C. irregularis sp. n. and C. derouxi grouped in the family Lynchellidae with their congeners to form the monophyletic genus Chlamydonella.
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FAM3A activates PI3K p110?/Akt signaling to ameliorate hepatic gluconeogenesis and lipogenesis.
Hepatology
PUBLISHED: 03-27-2014
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FAM3A belongs to a novel cytokine-like gene family, and its physiological role remains largely unknown. In our study, we found a marked reduction of FAM3A expression in the livers of db/db and high-fat diet (HFD)-induced diabetic mice. Hepatic overexpression of FAM3A markedly attenuated hyperglycemia, insulin resistance, and fatty liver with increased Akt (pAkt) signaling and repressed gluconeogenesis and lipogenesis in the livers of those mice. In contrast, small interfering RNA (siRNA)-mediated knockdown of hepatic FAM3A resulted in hyperglycemia with reduced pAkt levels and increased gluconeogenesis and lipogenesis in the livers of C57BL/6 mice. In vitro study revealed that FAM3A was mainly localized in the mitochondria, where it increases adenosine triphosphate (ATP) production and secretion in cultured hepatocytes. FAM3A activated Akt through the p110? catalytic subunit of PI3K in an insulin-independent manner. Blockade of P2 ATP receptors or downstream phospholipase C (PLC) and IP3R and removal of medium calcium all significantly reduced FAM3A-induced increase in cytosolic free Ca(2+) levels and attenuated FAM3A-mediated PI3K/Akt activation. Moreover, FAM3A-induced Akt activation was completely abolished by the inhibition of calmodulin (CaM).
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Changes in insecticide resistance of the rice striped stem borer (Lepidoptera: Crambidae).
J. Econ. Entomol.
PUBLISHED: 03-27-2014
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Application of insecticides is the most important method to control Chilo suppressalis (Walker) (Lepidoptera: Crambidae), and continuous use of individual insecticides has driven the rapid development of insecticide resistance in C. suppressalis during the past 30 yr. Monitoring insecticide resistance provides information essential for integrated pest management. Insecticide resistance of field populations to monosultap, triazophos, chlorpyrifos, and abamectin in China was examined in 2010 and 2011. The results indicated that the resistance levels of 14 field populations to four insecticides were significantly different. Four populations showed moderate resistance, and other populations possessed low-level resistance or were susceptible to monosultap. Nine populations displayed an extremely high or a high level of resistance to triazophos, whereas four populations were sensitive to this agent. Five populations exhibited a low level of resistance to abamectin, while the others remained sensitive. When compared with historical data, resistance to monosultap and triazophos decreased significantly, and the percentage of populations with high-level or extremely high-level resistance was obviously reduced. By contrast, the resistance to abamectin increased slightly. The increasing and decreasing resistance levels reported in this study highlight the different evolutionary patterns of insecticide resistance in C. suppressalis. An overreliance on one or two insecticides may promote rapid development of resistance. Slow development of resistance to abamectin, which was used mainly in mixtures with other insecticides, implies that the use of insecticide mixtures may be an effective method to delay the evolution of resistance to insecticides.
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Determination of glutathione in single HepG2 cells by capillary electrophoresis with reduced graphene oxide modified microelectrode.
Electrophoresis
PUBLISHED: 03-22-2014
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Determination of intracellular bioactive species will afford beneficial information related to cell metabolism, signal transduction, cell function, and disease treatment. In this study, the electrochemically reduced graphene oxide modified carbon fiber microdisk electrode (ER-GOME) was used as a detector of CZE-electrochemical detection and developed to detect glutathione (GSH). The electrocatalytic activity of the modified microelectrode was characterized by cyclic voltammetry. Under optimized experimental conditions, the concentration linear range of GSH was from 1 to 60 ?M. When the S/N ratio was 3, the concentration detection limit was 1 ?M. Compared with the unmodified carbon fiber microdisk electrode, the sensitivity was enhanced more than five times. With the use of this method, the average contents of GSH in single HepG2 cells were found to be 7.13 ± 1.11 fmol (n = 10). Compared with gold/mercury amalgam microelectrode, which was usually used in determining GSH, the electrochemically reduced graphene oxide modified carbon fiber microdisk electrode was friendly to environment for free mercury. Furthermore, there were several merits of the novel electrochemical detector coupled with CE, such as comparative repeatability, easy fabrication, and high sensitivity, hold great potential for the single-cell assay.
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Chicken biliary exosomes enhance CD4(+)T proliferation and inhibit ALV-J replication in liver.
Biochem. Cell Biol.
PUBLISHED: 03-14-2014
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Exosomes, which are small membrane vesicles of endocytic origin, carry lipids, RNA/miRNAs, and proteins and have immune modulatory functions. In this study, we isolated exosomes from the bile of specific pathogen-free chickens, 42-43 days of age, by using an ultracentrifugation and filtration method. The density of the exosomes, isolated by sucrose gradient fractionation, was between 1.13 and 1.19 g/mL. Electron microscopic observation of the liver showed that exosomes were present in the space of Disse and bile canaliculus. Chicken biliary exosomes displayed typical saucer-shaped, rounded morphology. Using liquid chromatography mass spectrum methodology, 196 proteins, including exosomal markers and several unique proteins, were identified and compared with mouse biliary exosomes. Noteworthy, CCCH type zinc finger antiviral protein was found on chicken biliary exosomes never described before. Furthermore, our data show that chicken biliary exosomes promote the proliferation of CD4(+) and CD8(+) T cells and monocytes from liver. In addition, chicken biliary exosomes significantly inhibit avian leukosis virus subgroup J, which is an oncogenic retrovirus, from replicating in the DF-1 cell line. These data indicate that chicken biliary exosomes possess the capacity to influence the immune responses of lymphocytes and inhibit avian leukosis virus subgroup J (ALV-J).
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The complete mitochondrial genome of the Parastromateus niger (Perciformes, Carangidae).
Mitochondrial DNA
PUBLISHED: 03-13-2014
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Abstract In this study, we have sequenced the complete mitochondrial genome of the Parastromateus niger. The mitochondrial genome is 16,561?bp long and contains 13 protein-coding genes, two rRNA genes, 22 tRNA genes and a control region. The gene order and composition of Parastromateus niger mitochondrial genome is similar to that of most other vertebrates. The nucleotide compositions of the light strand are 28.23% of A, 29.51% of C, 26.01% of T and 16.16% of G. With the exception of five tRNA genes, all other mitochondrial genes are encoded on the heavy strand.
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Metal Resistance-Related Genes are Differently Expressed in Response to Copper and Zinc Ion in Six Acidithiobacillus ferrooxidans Strains.
Curr. Microbiol.
PUBLISHED: 03-04-2014
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Metal resistance of acidophilic bacteria is very significant during bioleaching of copper ores since high concentration of metal is harmful to the growth of microorganisms. The resistance levels of six Acidithiobacillus ferrooxidans strains to 0.15 M copper and 0.2 M zinc were investigated, and eight metal resistance-related genes (afe-0022, afe-0326, afe-0329, afe-1143, afe-0602, afe-0603, afe-0604, and afe-1788) were sequenced and analyzed. The transcriptional expression levels of eight possible metal tolerance genes in six A. ferrooxidans strains exposed to 0.15 M Cu(2+) and 0.2 M Zn(2+) were determined by real-time quantitative PCR (RT-qPCR), respectively. The copper resistance levels of six A. ferrooxidans strains declined followed by DY26, DX5, DY15, GD-B, GD-0, and YTW. The zinc tolerance levels of six A. ferrooxidans strains exposed to 0.2 M Zn(2+) from high to low were YTW > GD-B > DY26 > GD-0 > DX5 > DY15. Seven metal tolerance-related genes all presented in the genome of six strains, except afe-0604. The metal resistance-related genes showed different transcriptional expression patterns in six A. ferrooxidans strains. The expression of gene afe-0326 and afe-0022 in six A. ferrooxidans strains in response to 0.15 M Cu(2+) showed the same trend with the resistance levels. The expression levels of genes afe-0602, afe-0603, afe-0604, and afe-1788 in six strains response to 0.2 M Zn(2+) did not show a clear correlation between the zinc tolerance levels of six strains. According to the results of RT-qPCR and bioinformatics analysis, the proteins encoded by afe-0022, afe-0326, afe-0329, and afe-1143 were related to Cu(2+) transport of A. ferrooxidans strains.
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Engineering uroporphyrinogen III methyltransferase as a red fluorescent reporter in E. coli.
Enzyme Microb. Technol.
PUBLISHED: 03-04-2014
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Uroporphyrinogen III methyltransferase (UMT) is a novel reporter owing to the catalytic products in the cells that emit strong red fluorescence under UV light. Here, we engineered the gene encoding the functional barley UMT (bUMT) by error-prone PCR and broadened the application UMT as a red fluorescent reporter in Escherichia coli. A variant, termed mbUMT, was selected and emitted stronger cell fluorescence than the wild type bUMT expressed in different E. coli strains, under different promoters and induction conditions respectively. The constructed mbUMT with a C-terminal ssrA tag was degraded in cells by the protease ClpXP encoded by E. coli chromosome, whereas the bUMT was expressed as active aggregates. Before they are exported to the periplasm, both proteins catalyze the substrate in the cytoplasm and emit cell fluorescence. The results suggested that the evolved bUMT is a better candidate to monitor in vivo degradation by E. coli ClpXP.
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The Regulatory Mechanism of Neurogenesis by IGF-1 in Adult Mice.
Mol. Neurobiol.
PUBLISHED: 02-24-2014
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Growth factors like insulin-like growth factor 1 (IGF-1) is reported to mediate neurogenesis in the subgranular zone (SGZ) and the subventricular zone (SVZ) of the adult mammalian brain, but its regulatory mechanism remains unclear. We generated transgenic mice overexpressing IGF-1 specifically in neural stem cells (NSCs) and assessed the effect of IGF-1 on neurogenesis in adult mice NSCs. Overexpression of IGF-1 could stimulate the expression of phospho-Akt and phospho-ERK1/2 while inducing proliferation and differentiation of NSCs in the SGZ and SVZ. The MEK/ERK inhibitor U0126 could inhibit ERK1/2 phosphorylation, further inhibiting the proliferation of NSCs in the SGZ and SVZ but had no effect on the phosphorylation of Akt. By contrast, The PI3K/Akt inhibitor LY294002 inhibited phosphorylation of Akt and differentiation of NSCs in the SGZ and SVZ, resulting in no change in the proliferation of NSCs and ERK1/2 phosphorylation. These results demonstrate that IGF-1 upregulates the proliferation of NSCs by triggering MEK/ERK pathway signaling in the adult mice SGZ and SVZ. Meanwhile, IGF-1 also induces differentiation of NSCs via the PI3K/Akt pathway in adult mice. However, we found no evidence of crosstalk between the PI3K/Akt and MEK/ERK pathways in adult mice NSCs. Our work provides new experimental evidence of the involvement of the PI3K/Akt and MEK/ERK pathways in the proliferation and differentiation of the NSCs of adult mice.
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FAM3A promotes vascular smooth muscle cell proliferation and migration and exacerbates neointima formation in rat artery after balloon injury.
J. Mol. Cell. Cardiol.
PUBLISHED: 02-19-2014
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The biological function of FAM3A, the first member of family with sequence similarity 3 (FAM3) gene family, remains largely unknown. This study aimed to determine its role in the proliferation and migration of vascular smooth muscle cells (VSMCs). Immunohistochemical staining revealed that FAM3A protein is expressed in the tunica media of rodent arteries, and its expression is reduced with an increase in prostaglandin E receptor 2 (EP2) expression after injury. In vitro, FAM3A overexpression promotes proliferation and migration of VSMCs, whereas FAM3A silencing inhibits these processes. In vivo, FAM3A overexpression results in exaggerated neointima formation of rat carotid artery after balloon injury. FAM3A activates Akt in a PI3K-dependent manner. In contrast, FAM3A induces ERK1/2 activation independent of PI3K. FAM3A protein is subcellularly located in mitochondria, where it affects ATP production and release. Activation of EP2 represses FAM3A expression, leading to impaired ATP production and release in VSMCs. FAM3A-induced activation of Akt and ERK1/2 pathways, proliferation and migration of VSMCs are inhibited by P2 receptor antagonist suramin. Furthermore, inhibition or knockdown of P2Y1 receptor inihibits FAM3A-induced proliferation and migration of VSMCs. In conclusion, FAM3A promotes proliferation and migration of VSMCs via P2Y1 receptor-mediated activation of Akt and ERK1/2 pathways. In injured vessels, FAM3A was repressed by upregulated EP2 expression, leading to the attenuation of ATP-P2Y1 receptor signaling, which is beneficial for preventing excessive proliferation and migration of VSMCs.
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Hydrodynamic tail vein injection of SOCS3 eukaryotic expression vector in vivo promoted liver lipid metabolism and hepatocyte apoptosis in mouse.
Biochem. Cell Biol.
PUBLISHED: 02-19-2014
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Suppressor of cytokine signaling 3 (SOCS3), a signal transduction cytokine, is involved in lipid metabolism as well as in cell proliferation, differentiation, apoptosis, and so on. To explore the effects of SOCS3 on apoptosis and lipid metabolism in liver, we used a simple effective method named hydrodynamic tail vein injection to overexpress SOCS3. Then orbital blood was obtained for the assessment of blood lipid after injection. Lipid metabolism related genes were detected by Western blot after the determination of serum lipids. Meanwhile, liver cell apoptosis was observed by Hoechst and TUNEL staining and the expression of apoptosis related proteins Bax, Bcl-2, and Caspase3 were detected as well as the JAK2/STAT3 signaling pathway. In addition, we also demonstrated the effect of SOCS3 in prime hepatocyte by overexpression or interference of SOCS3 along with SD1008, which is a specific inhibitor of the JAK2/STAT3 signaling pathway. Taken together, all the results indicated that SOCS3 promoted lipid synthesis in mice liver and promoted hepatocyte apoptosis by inhibiting the activation of the JAK2/STAT3 signaling pathway, however the detailed regulation mechanism had not yet been fully understood and needs further study.
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Gene expression profiling of the paracrine effects of uterine natural killer cells on human endometrial epithelial cells.
Int J Endocrinol
PUBLISHED: 02-18-2014
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The endometrium contains a population of immune cells that undergo changes during implantation and pregnancy. The majority of these cells are uterine natural killer (uNK) cells; however, it is unclear how these cells interact with endometrial epithelial cells. Therefore, we investigated the paracrine effects of the uNK cell-secretion medium on the gene expression profile of endometrial epithelial cells in vitro through microarray analysis. Our results, which were verified by qRT-PCR and western blot, revealed that soluble factors from uNK cells alter the gene expression profiles of epithelial cells. The upregulated genes included interleukin-15 (IL-15) and interleukin-15 receptor alpha (IL-15RA), which result in a loop that stimulates uNK cell proliferation. In addition, vascular endothelial growth factor C (VEGF-C) and chemokine (C-X-C motif) ligand 10 (CXCL-10) were also determined to be upregulated in epithelial cells, which suggests that uNK cells work synergistically with epithelial cells to support implantation and pregnancy. In addition, oriental herbal medicines have been used to treat infertility since ancient times; however, we failed to find that Zi Dan Yin can regulate these endometrial paracrine effects.
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Striatins contain a noncanonical coiled coil that binds protein phosphatase 2A A subunit to form a 2:2 heterotetrameric core of striatin-interacting phosphatase and kinase (STRIPAK) complex.
J. Biol. Chem.
PUBLISHED: 02-18-2014
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The protein phosphatase 2A (PP2A) and kinases such as germinal center kinase III (GCKIII) can interact with striatins to form a supramolecular complex called striatin-interacting phosphatase and kinase (STRIPAK) complex. Despite the fact that the STRIPAK complex regulates multiple cellular events, it remains only partially understood how this complex itself is assembled and regulated for differential biological functions. Our recent work revealed the activation mechanism of GCKIIIs by MO25, as well as how GCKIIIs heterodimerize with CCM3, a molecular bridge between GCKIII and striatins. Here we dissect the structural features of the coiled coil domain of striatin 3, a novel type of PP2A regulatory subunit that functions as a scaffold for the assembly of the STRIPAK complex. We have determined the crystal structure of a selenomethionine-labeled striatin 3 coiled coil domain, which shows it to assume a parallel dimeric but asymmetric conformation containing a large bend. This result combined with a number of biophysical analyses provide evidence that the coiled coil domain of striatin 3 and the PP2A A subunit form a stable core complex with a 2:2 stoichiometry. Structure-based mutational studies reveal that homodimerization of striatin 3 is essential for its interaction with PP2A and therefore assembly of the STRIPAK complex. Wild-type striatin 3 but not the mutants defective in PP2A binding strongly suppresses apoptosis of Jurkat cells induced by the GCKIII kinase MST3, most likely through a mechanism in which striatin recruits PP2A to negatively regulate the activation of MST3. Collectively, our work provides structural insights into the organization of the STRIPAK complex and will facilitate further functional studies.
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Nodal promotes the self-renewal of human colon cancer stem cells via an autocrine manner through Smad2/3 signaling pathway.
Biomed Res Int
PUBLISHED: 02-17-2014
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Colorectal cancer is one of the most common and fatal tumors. However, molecular mechanisms underlying carcinogenesis of colorectal cancer remain largely undefined. Here, we explored the expression and function of Nodal in colon cancer stem cells (CCSCs). Nodal and its receptors were present in numerous human colorectal cancer cell lines. NODAL and ALK-4 were coexpressed in human colon cancerous tissues, and NODAL, CD24, and CD44, markers for CCSCs, were expressed at higher levels in human colon cancerous tissues than adjacent noncancerous colon tissues. Human CCSCs were isolated by magnetic activated cell sorting using anti-CD24 and anti-CD44. Nodal transcript and protein were hardly detectable in CD44- or CD24-negative human colorectal cancer cell lines, whereas Nodal and its receptors were present in CCSCs. Notably, Nodal facilitated spheroid formation of human CCSCs, and phosphorylation of Smad2 and Smad3 was activated by Nodal in cells of spheres derived from human CCSCs. Collectively, these results suggest that Nodal promotes the self-renewal of human CCSCs and mediate carcinogenesis of human colorectal cancer via an autocrine manner through Smad2/3 pathway. This study provides a novel insight into molecular mechanisms controlling fate of human CCSCs and offers new targets for gene therapy of human colorectal cancer.
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Coupling exonuclease III with DNA metallization for amplified detection of biothiols at picomolar concentration.
Biosens Bioelectron
PUBLISHED: 02-14-2014
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For early diagnosis of diseases, the need of ultralow detection limit is an ongoing quest. In this work, by taking the uniqueness of Exonuclease III and DNA metallization, we demonstrate a facile turn-on fluorescent method for amplified detection of biothiols at picomolar concentration. This method relies on the amplification process achieved by the recycling of biothiols retrieved target DNA from silver depositions and the specific interactions between quadruplex and NMM. This method is simple in design, economic in operation and exhibits ultralow detection limit and excellent selectivity toward thiol-containing biomolecules among amino acids found in proteins and in serum samples. More importantly, the detection and discrimination process can be seen by the naked eye with the aid of an UV transilluminator. Therefore, this new concept may offer a potential approach for practical applications as an efficient biosensor for early detection of diseases.
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A 3-D artificial colon tissue mimic for the evaluation of nanoparticle-based drug delivery system.
Mol. Pharm.
PUBLISHED: 02-04-2014
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Functional engineered nanoparticles are promising drug delivery carriers. As the construction of a functional nanocarrier always needs the optimization of multiple technical variables, efficient in vitro high-throughput evaluation methods would help to shorten the development cycle. In the present study, we generated a tissue mimic of the colon of inflammatory bowel disease (IBD) patients. Generally, Caco-2 cells and THP-1 cells were grown in a 3-D matrix with different number, spatial distribution and specific extracellular cell matrix (ECM) composition according to real healthy and inflamed animal colon tissues. After interlerukin-1?/lipopolysaccharide (LPS) stimulation, the artificial model closely resembled the pathological features of IBD patient's colon, including massive cytokines and mucus production, epithelium defect and leukocytic infiltration. The tissue and cellular uptake of three different nanoparticles in the artificial model was similar to that in 2,4,6-trinitrobenzenesulfonic acid (TNBS) colitic mice. Most importantly, our artificial tissue can be placed into 96-well plates for high-throughput screening of drug delivery carriers for the treatment of IBD. Our study suggested a readily achievable way to improve current methodologies for the development of colon targeted drug delivery systems.
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The complete mitochondrial genome of the duskytail grouper Epinephelus bleekeri (Serranidae: Epinephelinae).
Mitochondrial DNA
PUBLISHED: 01-28-2014
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Abstract The complete mitochondrial genome of Epinephelus bleekeri was first presented in this study. It is 17,227?bp in length, consisted of 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and a non-coding control region. The overall base composition of the H-strand is 26.32% A, 28.65% T, 28.72% G and 16.31% C. With the exception of ND6 and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand. In particular, gene duplication is found on the tRNA-Asp gene, and five tandem repeat of this gene in E. bleekeri is different from most other vertebrates. The control region is rich in A?+?T (67.12%) and poor in G (14.00%), and tandem repeat sequences are observed in this non-coding region, indicating a useful marker for population genetic studies.
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In vitro and in vivo antioxidant activity of a fructan from the roots of Arctium lappa L.
Int. J. Biol. Macromol.
PUBLISHED: 01-26-2014
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To explore new antioxidant resource from food, a water-soluble polysaccharide (ALP1) was extracted and purified from the roots of Arctium lappa L. (A. lappa L.) through hot water extraction followed by ethanol precipitation, ion-exchange chromatography and gel filtration. The antioxidant activity of ALP1 was then evaluated in vitro and in vivo. ALP1 was characterized as a fructan composed of fructose and glucose in the ratio of 13.0:1.0, with an average molecular weight of 4600 Da. The linkages in ALP1 were ?1)-Fruf-(2?, Fruf-(2? and Glcp-(1?. In vitro antioxidant assays demonstrated that ALP1 possessed moderate ABTS(+) scavenging activity, strong hydroxyl radical scavenging activity and strong ferrous ion chelating activity. In in vivo antioxidant assays, ALP1 administration significantly enhanced antioxidant enzyme activities and total antioxidant capacity, as well as decreased the levels of malondialdehyde (MDA) in both the serum and liver of aging mice. These results suggest that ALP1 has potential as a novel natural antioxidant in food industry and pharmaceuticals.
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Insights into the paracrine effects of uterine natural killer cells.
Mol Med Rep
PUBLISHED: 01-22-2014
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Uterine natural killer (uNK) cells are recruited into the uterus during establishment of the implantation and placentation of the embryo, and are hypothesized to regulate uterine spiral artery remodeling and angiogenesis during the initial stages of pregnancy. Failures in uNK cell activation are linked to diseases associated with pregnancy. However, the manner in which these cells interact with the endometrium remain unknown. Therefore, this study investigated the paracrine effects of uNK cells on the gene expression profile of an endometrial epithelial and stromal cell co?culture system in vitro, using a microarray analysis. Results from reverse transcription?quantitative polymerase chain reaction and enzyme?linked immunosorbent assay experiments showed that soluble factors from uNK cells significantly alter endometrial gene expression. In conclusion, this study suggests that paracrine effects of uNK cells guide uNK cell proliferation, trophoblast migration, endometrial decidualization and angiogenesis, and maintain non?cytotoxicity of uNK cells.
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A peptide mimicking VGLL4 function acts as a YAP antagonist therapy against gastric cancer.
Cancer Cell
PUBLISHED: 01-16-2014
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The Hippo pathway has been implicated in suppressing tissue overgrowth and tumor formation by restricting the oncogenic activity of YAP. However, transcriptional regulators that inhibit YAP activity have not been well studied. Here, we uncover clinical importance for VGLL4 in gastric cancer suppression and find that VGLL4 directly competes with YAP for binding TEADs. Importantly, VGLL4's tandem Tondu domains are not only essential but also sufficient for its inhibitory activity toward YAP. A peptide mimicking this function of VGLL4 potently suppressed tumor growth in vitro and in vivo. These findings suggest that disruption of YAP-TEADs interaction by a VGLL4-mimicking peptide may be a promising therapeutic strategy against YAP-driven human cancers.
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Structure of MST2 SARAH domain provides insights into its interaction with RAPL.
J. Struct. Biol.
PUBLISHED: 01-13-2014
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The STE20 kinases MST1 and MST2 are key players in mammalian Hippo pathway. The SARAH domains of MST1/2 act as a platform to mediate homodimerization and hetero-interaction with a range of adaptors including RASSFs and Salvador, which also possess SARAH domains. Here, we determined the crystal structure of human MST2 SARAH domain, which forms an antiparallel homodimeric coiled coil. Structural comparison indicates that SARAH domains of different proteins may utilize a shared dimerization module to form homodimer or heterodimer. Structure-guided mutational study identified specific interface residues critical for MST2 homodimerization. MST2 mutations disrupting its homodimerization also impaired its hetero-interaction with RAPL (also named RASSF5 and NORE1), which is mediated by their SARAH domains. Further biochemical and cellular assays indicated that SARAH domain-mediated homodimerization and hetero-interaction with RAPL are required for full activation of MST2 and therefore apoptotic functions in T cells.
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Addition of pluronics® to reducible disulfide-bond-containing Pluronic®-PEI-SS specifically enhances circulation time in vivo and transfection efficiency in vitro.
J. Biomed. Mater. Res. Part B Appl. Biomater.
PUBLISHED: 01-04-2014
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To improve the circulation time and transfection efficiency of the polyplexes used in gene delivery, a series of Pluronic®/Pluronic®F127-PEI-SS/pDNA complexes (PFPS/pDNA), based on the addition of different kinds of Pluronics® to the reducible disulfide-bonds-containing Pluronic®F127-PEI-SS/pDNA (FPS/pDNA) polyplexes, was prepared and evaluated in Bcap and Hela cells in vitro and in vivo. The addition of Pluronics® with molecular weights and hydrophilic-lipophilic balance (HLBs) different from that in the FPS/pDNA complex resulted in five PFPS(1-5)/pDNA complexes, and the correlation between the structure of the free Pluronic® and the properties of the PFPS/pDNA complexes was investigated. The addition of Pluronics® resulted in slightly larger or same-sized nanoparticles of PFPS/pDNA at a constant N/P ratio. The PFPS copolymer displayed strong stability against DNase I digestion and serum degradation. PFPS-4 containing added Pluronic® L35, with an intermediate HLB of 19, showed a much higher transfection efficiency and less cytotoxicity than FPS or PEI-25 kDa in vitro. PFPS-4 also exhibited a considerably longer blood circulation time than FPS or PEI-25 kDa in vivo in mice, indicating that the addition of an intermediate Pluronic® can enhance the transfection efficiency of gene delivery systems.
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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.