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Find video protocols related to scientific articles indexed in Pubmed.
Subtemporal-anterior transtentoral approach to middle cranial fossa microsurgical anatomy.
J Craniofac Surg
PUBLISHED: 11-08-2014
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This study aimed to describe the topography of inferior and external dura mater of the middle cranial fossa through subtemporal-anterior transpetrosal approach and discuss the feasibility of improving the approach. Eight formalin-fixed adult cadaveric heads were studied, with the bones milled away in the lateral triangle region of the petrous bone, Kawase rhombus region, and inner triangle region of the petrous apex. The distances between the targets in these regions, as well as the angles after the dissection of zygomatic arch, were measured, and then the exposed petroclival and retrochiasmatic areas were observed under the microscope. There were significant variations in the distances between targets in the 3 milled regions among the specimens. After the dissection of zygomatic arch, the surgical view got an average increase of 12 degrees. The subtemporal anterior transpetrosal approach, as an improved subtemporal approach, can expose the lesions optimally, causing no injury to the hearing and reducing injuries to temporal lobe. On the other hand, the lateral bone of the petrous parts of the temporal bone is removed so as to improve the view to the retrochiasmatic area and expand the operative field.
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Spring Viremia of Carp Virus Induces Autophagy for Necessary Viral Replication.
Cell. Microbiol.
PUBLISHED: 10-21-2014
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Outbreaks of spring viremia of carp virus (SVCV) in several carp species and other cultivated fish can cause significant mortality and jeopardize the billion-dollar worldwide fish industry. SVCV, also known as Rhabdovirus carpio, is a bullet-shaped RNA virus that enters and amplifies in gill epithelium and later spreads to internal organs. Young fish under stressed conditions (spring cold water, etc.) are more vulnerable to SVCV-induced lethality due to their lack of a mature immune system. Currently, the host response of SVCV remains largely unknown. Here, we observed that autophagy is activated in SVCV-infected epithelioma papulosum cyprinid (EPC) cells. We demonstrated that the SVCV glycoprotein, rather than viral replication, activates the autophagy pathway. In addition, SVCV utilized the autophagy pathway to facilitate its own genomic RNA replication and to enhance its titers in the supernatants. Autophagy promoted the survival of SVCV-infected cells by eliminating damaged mitochondrial DNA generated during viral infection. We further showed that SVCV induces autophagy in EPC cells through the ERK/mTOR signaling pathway. Our results reveal a connection between autophagy and SVCV replication and propose autophagy suppression as a novel means to restrict SVCV viral replication.
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Clinical Course of Diabetes After Gastrectomy According to Type of Reconstruction in Patients with Concurrent Gastric Cancer and Type 2 Diabetes.
Obes Surg
PUBLISHED: 09-05-2014
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This study was conducted to evaluate course of diabetes after gastrectomy according to type of reconstruction performed for gastric cancer in patients with type 2 diabetes.
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Transformation of the education of health professionals in China: progress and challenges.
Lancet
PUBLISHED: 09-02-2014
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In this Review we examine the progress and challenges of China's ambitious 1998 reform of the world's largest health professional educational system. The reforms merged training institutions into universities and greatly expanded enrolment of health professionals. Positive achievements include an increase in the number of graduates to address human resources shortages, acceleration of production of diploma nurses to correct skill-mix imbalance, and priority for general practitioner training, especially of rural primary care workers. These developments have been accompanied by concerns: rapid expansion of the number of students without commensurate faculty strengthening, worries about dilution effect on quality, outdated curricular content, and ethical professionalism challenged by narrow technical training and growing admissions of students who did not express medicine as their first career choice. In this Review we underscore the importance of rebalance of the roles of health sciences institutions and government in educational policies and implementation. The imperative for reform is shown by a looming crisis of violence against health workers hypothesised as a result of many factors including deficient educational preparation and harmful profit-driven clinical practices.
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[Establishment of an allele-specific PCR method for direct screening of CYP21A2 gene mutation].
Zhonghua Yi Xue Yi Chuan Xue Za Zhi
PUBLISHED: 08-15-2014
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To establish an allele-specific PCR method for detect screening of CYP21A2 gene mutation.
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A home-built digital optical MRI console using high-speed serial links.
Magn Reson Med
PUBLISHED: 08-08-2014
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To develop a high performance, cost-effective digital optical console for scalable multichannel MRI.
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Molecular characterization, expression profiles of the ovine FSHR gene and its association with litter size.
Mol. Biol. Rep.
PUBLISHED: 08-08-2014
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Follicle-stimulating hormone receptor (FSHR) is a glycoprotein family member of G proteins coupled receptor super-family, and plays important roles in animal follicular development. In this study, we cloned the 5' flanking region of ovine FSHR gene and analyzed its genomic structure. One special domain named the LRRNT (Leucine rich repeat N-terminal domain) was found in the prediction amino acid sequence of ovine FSHR. RT-qPCR showed that ovine FSHR was expressed widely in detected tissues and was significantly higher in sexual gland (testicle and ovary) than in other tissues (heart, liver, spleen, lung, kidney, rumen, duodenum, muscle, fat, hypothalamus and pituitary) (P < 0.05). In addition, synonymous mutation g.-47C > T of the FSHR gene was detected and confirmed to be significantly associated with the litter size (P < 0.01), the genotype of CC had 0.42 (P < 0.01) and 0.53 (P < 0.01) lambs more than TC and TT genotype, respectively. Our results indicated the association of ovine FSHR with the litter size in sheep and FSHR could be used as a candidate gene for improving reproductive traits in industrial sheep breeding.
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The prognostic value of CD133 expression in non-small cell lung cancer: a meta-analysis.
Tumour Biol.
PUBLISHED: 06-29-2014
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CD133 has been identified as a potential cancer stem cell (CSC) marker in non-small cell lung cancer (NSCLC). However, the clinical and prognostic significance of CD133 in NSCLC remains controversial. In this study, a meta-analysis with a total number of 13 studies was performed to clarify the association between CD133 expression and clinical outcomes in publications up to June 2013. Odds ratios (ORs) and their 95 % confidence intervals (CIs) were used to assess the association between CD133 expression and the clinicopathological characteristics of NSCLC. Hazard ratios (HRs) and their 95 % CI were used to quantify the predictive ability of CD133 on NSCLC prognosis. Analysis of these data showed that CD133 expression was not associated with any clinicopathological parameters except for histology (pooled OR?=?1.35, 95%CI?=?1.04-1.76, P?=?0.024) and tumor differentiation (pooled OR?=?3.19, 95%CI?=?1.10-9.21, P?=?0.032). Simultaneously, we also found that positive CD133 expression was not associated with disease-free survival (DFS) (pooled HR?=?1.76, 95 % CI?=?0.87-3.57, P?=?0.114, random-effect) but was associated with overall survival (OS) (pooled HR?=?2.06, 95 % CI?=?1.08-3.91, P?=?0.027, random-effect). Overall, it is appropriate to regard CD133 expression as a potential prognostic factor for the OS of NSCLC patients.
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Irradiation facilitates the inhibitory effect of the heat shock protein 90 inhibitor NVP-BEP800 on the proliferation of malignant glioblastoma cells through attenuation of the upregulation of heat shock protein 70.
Exp Ther Med
PUBLISHED: 06-23-2014
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The present study aimed to investigate the effect of NVP-BEP800, a novel heat shock protein (Hsp) 90 inhibitor of the 2-aminothieno[2,3-d]pyrimidine class, in combination with radiation on glioblastoma cells. T98G human glioblastoma cells were treated with dimethyl sulfoxide (DMSO), NVP-BEP800, NVP-BEP800 in combination with X-ray irradiation (10 Gy, 20 min), or X-ray irradiation only, and cultured for 40 h. Cell viability was measured upon completion of the treatments. In addition, apoptosis was measured and immunoblot analysis was performed to analyze the expression levels of cellular protein inhibitory ?B kinase ? (IKK?). The combined treatment with NVP-BEP800 and X-ray irradiation resulted in the synergistic destruction of malignant cells. Furthermore, NVP-BEP800 significantly induced apoptosis in the human glioblastoma cells. The immunoblot analysis data indicated that NVP-BEP800 markedly reduced the expression level of IKK?. The results also revealed that X-ray irradiation significantly attenuated the increase in the level of Hsp70 in cells treated with NVP-BEP800. Since elevated levels of Hsp70 are associated with drug resistance induced by Hsp90 inhibitors, the effects of X-ray irradiation on Hsp70 levels may be associated with the enhanced effect on cells of the presence of irradiation. The results of the current study suggest that irradiation enhances the inhibitory effect of NVP-BEP800 on the proliferation of malignant glioblastoma cells by downregulating the expression level of cellular signaling protein IKK? and attenuating the upregulation of Hsp70 that is induced by NVP-BEP800.
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A Comprehensive Transcriptome Provides Candidate Genes for Sex Determination/Differentiation and SSR/SNP Markers in Yellow Catfish.
Mar. Biotechnol.
PUBLISHED: 06-04-2014
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Sex dimorphic growth pattern has significant theory and application implications in fish. Recently, a Y- and X-specific allele marker-assisted sex control technique has been developed for mass production of all-male population in yellow catfish (Pelteobagrus fulvidraco), but the genetic information for sex determination and sex control breeding has remained unclear. Here, we attempted to provide the first insight into a comprehensive transcriptome covering multiple tissues from XX females, XY males, and YY super-males of yellow catfish by using 454 GS-FLX platform, for a better assembly and gene coverage. A total of 1,202,933 high quality reads (about 540 Mbp) were obtained and assembled into 28,297 contigs and 141,951 singletons. BLASTX searches against the NCBI non-redundant protein database (nr) led a total of 52,564 unique sequences including 18,748 contigs and 33,816 singletons to match 25,669 known or predicted unique proteins. All of them with annotated function were categorized by gene ontology (GO) analysis, and 712 were assigned to reproduction and reproductive process. Some potential genes relevant to reproductive system including steroid hormone biosynthesis and GnRH (gonadotropin-releasing hormone) signaling pathway were further identified by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis; and at least 21 sex determination and differentiation-related genes, such as Dmrt1, Sox9a/b, Cyp19b, WT1, and AMH were identified and characterized. Additionally, a total of 82,794 simple sequence repeats (SSRs), 26,450 single nucleotide polymorphisms (SNPs), and 4,145 insertions and deletions (INDELs) were revealed from the transcriptome data. Therefore, the current transcriptome resources highlight further studies on sex-control breeding in yellow catfish and will benefit future studies on reproduction and sex determination in teleost fish.
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Chemical composition, crystal structure, and their relationships with the intrinsic properties of spinel-type crystals based on bond valences.
Inorg Chem
PUBLISHED: 05-28-2014
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Spinel-type crystals may possess complex and versatile chemical composition and crystal structure, which leads to difficulty in constructing relationships among the chemical composition, crystal structure, and intrinsic properties. In this work, we develop new empirical methods based on bond valences to estimate the intrinsic properties, namely, compressibility and thermal expansion of complex spinel-type crystals. The composition-weighted average of bond force constants in tetrahedral and octahedral coordination polyhedra is derived as a function of the composition-weighted average of bond valences, which can be calculated according to the experimental chemical composition and crystal structural parameters. We discuss the coupled effects of tetrahedral and octahedral frameworks on the aforementioned intrinsic properties. The bulk modulus could be quantitatively calculated from the composition-weighted average of bond force constants in tetrahedral and octahedral coordination polyhedra. In contrast, a quantitative estimation of the thermal expansion coefficient could be obtained from the composition-weighted average of bond force constants in octahedral coordination polyhedra. These empirical methods have been validated by the results obtained for a new complex quaternary spinel-type oxynitride Mg0.268Al2.577O3.733N0.267 as well as MgAl2O4 and Al2.85O3.45N0.55 from the literature. Further, these empirical methods have the potential to be extensively applied in other types of complex crystals.
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Quantitation of P450 3A4 endogenous biomarker - 4?-hydroxycholesterol - in human plasma using LC/ESI-MS/MS.
Biomed. Chromatogr.
PUBLISHED: 05-28-2014
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4?-Hydroxycholesterol (4?-HC) has been proposed as a new endogenous biomarker for cytochrome P450 3A4/5 activity. Therefore, it is important to have a robust method for its accurate determination in human plasma. Here a liquid chromatography-tandem mass spectrometry with electrospray ionization (LC/ESI-MS/MS) assay for the quantitation of 4?-HC in human plasma is described. While the calibration standards were prepared in a surrogate matrix for human plasma, the quality control samples were prepared in human plasma to mimic the incurred study samples. In order to achieve accurate determination of 4?-HC, the chromatographic separation of 4?-HC from its isomers, especially 4?-hydroxycholesterol (4?-HC), was crucial. In the absence of an authentic 4?-HC standard at the time of this study, an alternative selectivity test strategy was developed to confirm the separation. After being alkalized with potassium hydroxide, the human plasma sample (50 ?L) was extracted with hexane, derivatized into picolinyl esters using picolinic acid, extracted again with hexane, and then analyzed by LC/ESI-MS/MS. The calibration curve range was 5-500 ng/mL and the chromatographic separation was achieved on a 50 × 2.1 mm Thermal Hypersil Gold column with a gradient elution. The assay accuracy, precision, linearity, selectivity and analyte stability throughout the analysis were established. The validated assay was successfully applied to a Phase I clinical study for the measurement of 4?-HC in human plasma.
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Novel endosomolytic poly(amido amine) polymer conjugates for systemic delivery of siRNA to hepatocytes in rodents and nonhuman primates.
Bioconjug. Chem.
PUBLISHED: 05-05-2014
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The application of small interfering (si)RNAs as potential therapeutic agents requires safe and effective methods for their delivery to the cytoplasm of the target cells and tissues. Recent studies have shown significant progress in the development of targeting reagents that facilitate the recognition of, and siRNA delivery to, specific cell types. Among recently reported delivery approaches, polymers with amphipathic properties have been used to enable endosome escape and cytosolic delivery. Here, we describe a linear amphipathic poly(amido amine) polymer conjugate system for the efficient siRNA delivery in vitro and in vivo. This polymer contains a novel amine bearing bis-acrylamide monomer designed for increasing amine density, which resulted in substantial improvement in liver uptake and RNAi activity compared to our previously reported poly(amido amine disulfide) polymer.1 The activity for this liver targeted delivery system was demonstrated in rodents and nonhuman primates.
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Oligochitosan stimulated phagocytic activity of macrophages from blunt snout bream (Megalobrama amblycephala) associated with respiratory burst coupled with nitric oxide production.
Dev. Comp. Immunol.
PUBLISHED: 04-08-2014
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The immunostimulating effects of oligochitosan have been proven in several fish, however, the mechanisms underlying the stimulation are not characterized. In the present study, the effects of oligochitosan were investigated using macrophages isolated from blunt snout bream (Megalobrama amblycephala). The results showed that the phagocytic activity of the macrophages was enhanced by the addition of oligochitosan in vitro and in vivo. The two of the most important antimicrobial pathways of macrophages, NADPH oxidase and iNOS pathways were included for further studies. The amounts of superoxide anion and the mRNAs of the five subunits of NADPH oxidase genes were significantly enhanced in the oligochitosan-treated macrophages and macrophages isolated from fish fed with feed containing oligochitosan. In addition, the NO production, iNOS activity and iNOS gene expression were all significantly increased in the presence of oligochitosan. Furthermore, the mRNA levels of the TNF-? and IL-1? were also significantly increased in the macrophages derived from fish fed with oligochitosan. In conclusion, the stimulation effects of oligochitosan on the phagocytic activity of the fish macrophages were associated with respiratory burst coupled with nitric oxide production.
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Molecular cloning and expression analysis of immunoglobulin M heavy chain gene of blunt snout bream (Megalobrama amblycephala).
Fish Shellfish Immunol.
PUBLISHED: 03-25-2014
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Immunoglobulins (Igs), which bind antigens with high specificity, are essential molecules in adaptive immune system of jawed vertebrates. In this study, cDNA encoding the secreted form of the immunoglobulin heavy chain of IgM (sIgM) was cloned from the mesonephros of blunt snout bream (Megalabrama amblycephala) using RT-PCR and rapid amplification of cDNA ends (RACE). The full-length cDNA of sIgM heavy chain gene has 1961 nucleotides encoding a putative protein of 569 amino acids, constant region shares high amino acid identity with that of Ctenopharyngodon idella (80%), Carassius auratus langsdorfii (65%) and Danio rerio (59%). Multiple protein sequence alignment revealed that blunt snout bream sIgM was clustered with the homologues of cyprinid fish and constructed one clade. Using quantitative real-time PCR (qRT-PCR) analysis, the level of sIgM mRNA was determined, with a V-shape change pattern: decreased initially from unfertilized egg stage to 4 cells stage and increased from 16 cells stage to prelarva. This sharp drop indicates that sIgM mRNA is maternally transferred, and was continuously degraded until 16 cells stage. The drastic rising in sIgM level from blastula stage to prelarva might be attributed to embryonic stem cell differentiation procedure. Compared with juvenile fish, the expression of sIgM was significantly higher in pronephros, liver, spleen, gill and muscle of adult fish. After the injection of Aeromonas hydrophila, the expression pattern of sIgM was found first down-regulated at 4 h, then up-regulated and reached the peak at 7 d and 21 d in mesonephros, spleen, liver and gill, respectively.
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Sequence analysis and expression regulation of rbp4 by 9-cis-RA in Megalobrama amblycephala.
Fish Physiol. Biochem.
PUBLISHED: 03-24-2014
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Retinol-binding protein 4 (rbp4) is mainly synthesized in the liver, where it binds retinol and then enters the bloodstream, delivering retinol to cells. The full-length cDNA coding rbp4 was cloned from Megalobrama amblycephala. The amino acid sequence showed strong homology with the homologues of other vertebrates, and all structural and functional domains were highly conserved. The mRNA levels in different tissues and development stages detected by quantitative real-time PCR revealed that M. amblycephala rbp4 was highly expressed in liver (P < 0.001), but the lower levels were also detected in eyes, kidney, intestine, and spleen. During the different development stages, the rbp4 mRNA appeared until 28 hours post-fertilization (hpf), underwent a slight drop, and then gradually increased after 50 hpf. In addition, the promoter sequence of M. amblycephala rbp4 was obtained using thermal asymmetric interlaced PCR. Two single nucleotide polymorphism sites (-385A>G and -329C>T) were found in the promoter. Transfection with recombinant plasmids of two different haplotypes (GT, AC) showed that 9-cis-retinoic acid (RA) increased the promoter activity, but the AC haplotype was more sensitive to RA.
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Temozolomide induces autophagy via ATM?AMPK?ULK1 pathways in glioma.
Mol Med Rep
PUBLISHED: 03-18-2014
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Autophagy is a cytoprotective process, which occurs following temozolomide (TMZ) treatment, and contributes to glioma chemoresistance and TMZ treatment failure. However, the molecular mechanisms by which TMZ induces autophagy are largely unknown. In the current study, the ataxia?telangiectasia mutated (ATM) inhibitor KU?55933, adenosine monophosphate?activated protein kinase (AMPK) inhibitor compound C, and U87MG and U251 cell lines were employed to investigate the molecular mechanisms of TMZ?induced autophagy in glioma, and to evaluate the effects of autophagy inhibition on TMZ cytotoxicity. KU?55933 and compound C were observed to inhibit the activation of autophagy?initiating kinase ULK1 and result in a significant decrease of autophagy as indicated by depressed LC3B cleavage and acidic vesicular organelle formation. The activation of AMPK?ULK1 was ATM dependent. Autophagy inhibition via the AMPK inhibitor compound C augmented TMZ cytotoxicity as observed by depressed cell viability, increased ?H2AX?marked double?strand breaks (DSBs) and elevated numbers of apoptotic glioma cells. In conclusion, TMZ induced autophagy via ATM?AMPK?ULK1 pathways. TMZ chemoresistance may therefore be overwhelmed by targeting AMPK, particularly for the treatment of O6?methylguanine DNA methyltransferase?negative gliomas.
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A prospective, randomized, controlled study of ?-3 fish oil fat emulsion-based parenteral nutrition for patients following surgical resection of gastric tumors.
Nutr J
PUBLISHED: 03-17-2014
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Nutrients such as ?-3 fatty acids including fish oil components eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) suppress the growth and promote apoptosis of tumor cells, improve immune function and reduce the effects of systemic inflammatory response syndrome. We sought to investigate the effect of ?-3 fish oil fat emulsion-based parenteral nutrition (PN) on nutritional state, immune function, inflammatory reaction, expression of tumor factors and complication incidence in patients after surgical resection of gastric cancer.
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Characterization of muscle morphology and satellite cells, and expression of muscle-related genes in skeletal muscle of juvenile and adult Megalobrama amblycephala.
Micron
PUBLISHED: 03-14-2014
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Hyperplasia and hypertrophy are two distinct processes of skeletal muscle growth regulated by four myogenic regulatory factors (MRFs, contains MyoD, Myf5, Mrf4 and myogenin (MyoG)) and myostatin (MSTN). In this study, characterization of muscle morphology and satellite cells in juvenile (1-year-old) and adult (2-year-old) Megalobrama amblycephala was described. Compared with 1-year-old M. amblycephala, the diameter scope of epaxial, horizontal septum and hypaxial muscle fibers including red and white muscle in 2-year-old fish exhibited broader with dramatic reduction in frequency distribution of <20 ?m diameter, nevertheless observable increase in frequency distribution of >50 ?m diameter. Intermyofibrillar (IM) nuclei were also found except numerous subsarcolemmal (SS) nuclei in 2-year-old fish, whereas only SS nuclei were observed in 1-year-old fish. Immunofluorescence results showed that more satellite cells existed in red muscle than white muscle in 1-year-old fish, rather than 2-year-old fish. Moreover, we observed predominant increase in the mRNA levels of MyoD, Myf5, Mrf4, and MSTN during muscle development of fish in 2-year-old fish except MyoG.
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Molecular cloning and expression analysis of mannose receptor in blunt snout bream (Megalobrama amblycephala).
Mol. Biol. Rep.
PUBLISHED: 03-10-2014
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Mannose receptor (MR) plays a significant role in innate immune responses to pathogens in vertebrates. Here we characterized the first teleost MR from Megalobrama amblycephala, named maMR and its expression patterns were investigated. The full-length maMR consists of 5,295 bp encoding a putative protein of 1,433 amino acids. The predicted amino acid sequences showed that maMR contained a signal peptide, a cysteine-rich domain, a single fibronectin type II domain, eight tandemly arranged C-type lectin-like domains, a transmembrane domain and a C-terminal cytoplasmic domain. Phylogenetic analysis revealed the highest similarity of maMR with Danio rerio MR predicted by computational analysis. The maMR-mRNAs were ubiquitously transcribed in different tissues, However the highest transcripts were observed in head kidney. Transcripts of maMR significantly increased at the late stages of embryo and continued to be at the high levels after hatching. The maMR transcripts were significantly increased in M. amblycephala after stimulation with killed Aeromonas hydrophila.
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Pathogenesis analysis of pituitary adenoma based on gene expression profiling.
Oncol Lett
PUBLISHED: 02-13-2014
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The aim of the current study was to investigate the pathogenesis of pituitary adenoma through screening of the differentially-expressed genes (DEGs) and proteins in normal pituitary and pituitary adenoma tissues, and analyzing the interactions among them. Following the acquisition of gene expression profiling data from a public functional genomics data repository, Gene Expression Omnibus, DEGs were screened in normal pituitary and pituitary adenoma tissues. Upregulated and downregulated DEGs were further identified through gene ontology functional enrichment analysis. Subsequently, the DEGs were mapped to the Search Tool for the Retrieval of Interacting Genes database, and the protein-protein interaction (PPI) networks of the upregulated and downregulated DEGs were constructed. Finally, the functional modules of the PPI network of the downregulated DEGs were analyzed. In total, 211 upregulated and 413 downregulated DEGs were screened between the normal pituitary and pituitary adenoma samples. Downregulated DEGs were associated with certain functions, including the immune response, hormone regulation and cell proliferation. Upregulated genes were associated with cation transport functions. Five modules were acquired from the PPI network of the downregulated DEGs. Transcription factors, including signal transducer and activator of transcription 3 (STAT3), interleukin 6 (IL-6), B-cell lymphoma 6 protein, early growth response 1, POU1F1, jun B proto-oncogene and FOS were the core nodes in the functional modules. In summary, the DEGs and proteins were identified through screening gene expression profiling and PPI networks. The results of the present study indicated that low expression levels of hormone- and immune-related genes facilitated the occurrence of pituitary adenoma. Low expression levels of IL-6 and STAT3 were significant in the dysimmunity of pituitary adenoma. Furthermore, the low expression level of POU1F1 contributed to the reduction in pituitary hormone secretion.
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Characterization of a novel CC chemokine CCL4 in immune response induced by nitrite and its expression differences among three populations of Megalobrama amblycephala.
Fish Shellfish Immunol.
PUBLISHED: 02-10-2014
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A novel CC chemokine gene, chemokine CC motif ligand 4 (CCL4), was isolated from Megalobrama amblycephala. The full-length cDNA was 913 bp, encoding 94 amino acid residues. The deduced amino acid sequence possessed the typical arrangement of four cysteines as found in other known CC chemokines. The expression of M. amblycephala CCL4 during the early development showed the mRNA levels before hatching and at 62 h post fertilized (hpf) were significantly higher than other post-hatching stages (P < 0.05). Besides, it was widely expressed in all detected tissues with the highest transcription in liver, followed by intestine, spleen and gill, where a larger number of immune cells including lymphocytes and macrophages are present. Our findings had fully confirmed that CCL4 expression was strongly induced in vitro and quickly up-regulated after nitrite stress, then substantially altered in all tested tissues, supporting a potential pro-inflammatory function. We also indicated that inflammation effect might firstly happen in blood after nitrite stress. Furthermore, the tissue expression differences of CCL4 among three natural populations revealed that CCL4 mRNA in Yuni Lake population was obviously higher than the other two populations, Liangzi Lake population and Poyang Lake population, which will provide valuable insights into breeding strategies for selecting population with better immune property of M. amblycephala.
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Development of a new compact intraoperative magnetic resonance imaging system: concept and initial experience.
Neurosurgery
PUBLISHED: 01-31-2014
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Magnetic resonance imaging (MRI) during surgery has been shown to improve surgical outcomes, but the current intraoperative MRI systems are too large to install in standard operating suites. Although 1 compact system is available, its imaging quality is not ideal.
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The complete mitochondrial genome of the hybrid of Megalobrama amblycephala (?)?×?Parabramis pekinesis (?).
Mitochondrial DNA
PUBLISHED: 01-24-2014
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Abstract The complete mitochondrial genome sequence of the hybrid of blunt snout bream (Megalobrama amblycephala) (?)?×?white Amur bream (Parabramis pekinesis) (?) (M?×?P) was determined using the PCR-based method for the first time. The total length of the mitogenome was 16,623?bp in accordance with blunt snout bream. It contained 37 genes (13 protein-coding genes, 22 transfer RNA genes and 2 ribosomal RNA genes) and 2 main non-coding regions (the control region and the origin of the light strand replication). Compared with the complete mitochondrial genome of the parents, results showed the hybrid bream was consistent with a maternal inheritance; however, we also found a termination codon mutation in Cytb and a double-based site in 16S rRNA which was species-specific for the male parent P. pekinesis. The complete mitochondrial genome sequence of the M?×?P bream provided an important data set for further study in mitochondrial inheritance mechanism.
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The complete mitochondrial genome sequence of Parabramis pekinensis strenosoma (Cypriniformes: Cyprinidae).
Mitochondrial DNA
PUBLISHED: 01-21-2014
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Abstract Parabramis pekinensis strenosoma belongs to the family Cyprinidae. In the present study, we obtain the complete mitochondrial genome of P. pekinensis strenosoma by PCR amplification and DNA sequencing. It is a circular double-stranded DNA molecule of 16,623 base pairs in length, consisting of the typical structure of 22 transfer RNA genes, 13 protein-coding genes, 2 ribosomal RNA genes, as well as 2 main non-coding regions (the control region and the origin of the light strand replication). Compared with Parabramis pekinensis, the two different subspecies share 99.58% nucleotide sequence similarity and the biggest nucleotide sequence discrepancy between homologous genes are observed in ND2 for protein-coding genes and in tRNA-Ala for tRNA-coding genes. The complete mitochondrial genome sequence data are of great use for phylogenetic analysis and studies of population genetics and germplasm resources of P. pekinensis strenosoma.
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The complete mitochondrial genome of natural Paramisgurnus dabryanus (Cypriniformes: Cobitidae).
Mitochondrial DNA
PUBLISHED: 01-14-2014
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Abstract Paramisgurnus dabryanus, a small-sized freshwater fish species, is one of the most important cultured fish in East Asia from Russia to China. In this study, the complete mitochondrial genome of P. dabryanus is sequenced to be 16,566?bp in length, including 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, a control region and the origin of the light strand replication. The overall base composition of P. dabryanus in descending order is A 29.66 %, T 28.04%, C 25.81% and G 16.49%, with a slight A?+?T bias. The mitogenome sequence data may provide useful information to the population genetics analysis of P. dabryanus and the elucidation of evolutionary mechanisms in Cobitidae.
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Basal and benzo[a]pyrene-induced expression profile of phase I and II enzymes and ABC transporter mRNA in the early life stage of Chinese rare minnows (Gobiocypris rarus).
Ecotoxicol. Environ. Saf.
PUBLISHED: 01-13-2014
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ATP-binding cassette (ABC) transporters together with phase I and II detoxification enzymes have been considered as included in a cellular detoxification system. Previous studies have highlighted the involvement of aryl hydrocarbon receptor (AHR) and Cyp1a in PAH-induced embryo toxicity. However, the response of other xenobiotic enzymes/transporters in PAH-mediated embryo toxicity is not fully characterized. In the present study, rare minnow embryos were exposed to 10 and 100µg/L BaP within 4h post-fertilization (hpf) up to 168 hpf. RNA was extracted at 24, 48, 96, and 168 hpf. The basal and BaP-induced expression of phase I enzyme genes (cyp1a, 1b1, and 1c1), phase II enzyme gene (gstm and ugt1a), and ABC transporter genes (abcb1, abcc1, abcc2, and abcg2) mRNA was determined using real-time PCR. Severe developmental defects (e.g., spinal deformities, pericardial and yolk-sac edema) were observed in the BaP treated groups. The basal expression showed that gstm was most strongly expressed, followed by abcb1, ugt1a, and abcc2, whereas cyp1a, 1b1, 1c1, and abcg2 showed weak expression. BaP significantly induced the mRNA expression of three CYP1s (cyp1a, 1b1, and 1c1) (p<0.05) and the ABC transporters (abcc1, abcc2, and abcg2) in a dose-dependent manner. However, the mRNA expression of Phase II enzymes (gstm, ugt1a) for the BaP treatments showed no significant difference with that of the controls. Furthermore, distinct induced patterns of these genes were observed during different exposure periods. Simultaneous up-regulation of the cyp and ABC transporter gene transcripts suggests that a possible involvement and cooperation in the detoxification process could provide protection against the BaP toxicity of rare minnows at the early life stage.
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Molecular characterization and expression patterns of myogenin in compensatory growth of Megalobrama amblycephala.
Comp. Biochem. Physiol. B, Biochem. Mol. Biol.
PUBLISHED: 01-05-2014
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Myogenin (myog) is a muscle-specific basic helix-loop-helix (bHLH) transcription factor that plays an essential role in regulating skeletal muscle development and growth. To investigate molecular characterization of myog and the effect of starvation/refeeding on the gene expression, we isolated the myog cDNA sequence and analyzed the expression patterns using quantitative real-time polymerase chain reaction in Megalobrama amblycephala. Sequence analysis indicated that M. amblycephala myog shared an analogous structure with the highly conserved His/Cys-rich, bHLH and C-terminal helix III domains with other vertebrates. Sequence alignment and phylogenetic tree showed that M. amblycephala myog had the highest identity with the homologues of Ctenopharyngodon idella and Cyprinus carpio. Spatio-temporal expression patterns revealed that myog mRNA levels at the segmentation period and 12 h post-hatching (hph) were significantly higher than at other development stages (P<0.05). Furthermore, the highest myog expression level was predominantly observed in white muscle compared with the other types of muscle. Fish body weight continuously decreased during 21-day starvation and then significantly increased after 7days of refeeding and reached the similar level to the control at 21days of refeeding, indicating that the pattern of complete compensatory growth possibly occurred in M. amblycephala; meanwhile, the relative somatic growth rate after refeeding was also dramatically higher than the control group. In addition, the myog expression decreased during 21days of starvation and then exhibited a strong rebound effect after 7days of refeeding and subsequently declined gradually to the control level by 21days of refeeding.
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Expression of Ghrelin in gastrointestinal tract and the effect of early weaning on Ghrelin expression in lambs.
Mol. Biol. Rep.
PUBLISHED: 01-03-2014
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Ghrelin, an endogenous ligand for growth hormone secretagogue receptor, plays an important role in stimulating hormone secretion, development of gastrointestinal tract, food intake and regulating energy balance of animals. In this study we isolated the cDNA sequence of ovine Ghrelin from the abomasums of 7-day-aged lambs. Real-time PCR was used to determine the abundance of Ghrelin mRNA in lamb gastrointestinal tract, and analyze the development changes of abomasums Ghrelin mRNA expression in 0-56 days lambs, as well as find the effects of 42-day weaning on Ghrelin mRNA expression in lamb abomasums. The results showed that: (1) Ghrelin mRNA was expressed widely in gastrointestinal tract and was significantly higher in the abomasums than in other tissues (rumen, reticulum, omasum, duodenum, jejunum, ileum) (P < 0.01); (2) The expression of abomasums Ghrelin mRNA in lamb increased with the growth of age, it reached a plateau at the age of 49 days, however, got a slightly decrease at the age of 56 days; (3) The expression of abomasums Ghrelin mRNA of the 42 days-weaned groups were significantly lower than the no-weaned groups (P < 0.05), and the Ghrelin mRNA expression of the two treatments reached a maximum at the age of 49 days; (4) Correlation analysis indicated that the linear correlativity between abomasums Ghrelin mRNA expression and abomasums weight was very prominent (R(2) = 0.647, P = 0.009). Our results suggested that ovine Ghrelin gene may play an important role in the development of lamb abomasums and 42-day weaning could down regulate the expression of abomasum Ghrelin mRNA, but the mechanism of these needs further research.
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Sex-biased miRNAs in gonad and their potential roles for testis development in yellow catfish.
PLoS ONE
PUBLISHED: 01-01-2014
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Recently, YY super-male yellow catfish had been created by hormonal-induced sex reversal and sex-linked markers, which provides a promising research model for fish sex differentiation and gonad development, especially for testis development. MicroRNAs (miRNAs) have been revealed to play crucial roles in the gene regulation and gonad development in vertebrates. In this study, three small RNA libraries constructed from gonad tissues of XX female, XY male and YY super-male yellow catfish were sequenced. The sequencing data generated a total of 384 conserved miRNAs and 113 potential novel miRNAs, among which 23, 30 and 14 miRNAs were specifically detected in XX ovary, XY testis, and YY testis, respectively. We observed relative lower expression of several miR-200 family members, including miR-141 and miR-429 in YY testis compared with XY testis. Histological analysis indicated a higher degree of testis maturity in YY super-males compared with XY males, as shown by larger spermatogenic cyst, more spermatids and fewer spermatocytes in the spermatogenic cyst. Moreover, five miR-200 family members were significantly up-regulated in testis when treated by 17?-ethinylestradiol (EE2), high dose of which will impair testis development and cell proliferation. The down-regulation of miR-141 and 429 coincides with the progression of testis development in both yellow catfish and human. At last, the expression pattern of nine arbitrarily selected miRNAs detected by quantitative RT-PCR was consistent with the Solexa sequencing results. Our study provides a comprehensive miRNA transcriptome analysis for gonad of yellow catfish with different sex genotypes, and identifies a number of sex-biased miRNAs, some of that are potentially involved in testis development and spermatogenesis.
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Transplantation of bone marrow stromal cells enhances infiltration and survival of CNP and Schwann cells to promote axonal sprouting following complete transection of spinal cord in adult rats.
Am J Transl Res
PUBLISHED: 01-01-2014
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This study aimed to investigate the roles of bone marrow stromal cells (BMSCs) in promoting axonal regeneration after complete transection of spinal cord in adult rats. Transplantation was done 9 days after injury. Only a few BMSCs were detected at the injury site 8 weeks after transplantation, yet there was robust growth of axons. The scarcity of surviving BMSCs may attribute to the adverse conditions in their ambient environment. In this connection, the immediate accumulation of a large number of macrophages/reactive microglia following BMSCs transplantation and subsequent cavitation of tissues may be detrimental to their survival. An unexpected finding following BMSCs transplantation was the marked increase in the nestin, GFAP, NF200, olig 3 and CNP positive cells at the injury site. Immunoelectron microscopy showed CNP cells were oval or fibroblast-like and had multiple perineurial-like compartments with long extending filopodia. The spatial relationship between regenerating axons and CNP-positive cells was also confirmed by double immunofluorescence staining. Our results suggest that transplantation of BMSCs elicits the influx and survival of local cells including CNP positive cells and Schwann cells into injury site, which provide structural support for the axon regeneration and remyelination after spinal cord injury.
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A novel high-power dual-band coupled-line Gysel power divider with impedance-transforming functions.
ScientificWorldJournal
PUBLISHED: 01-01-2014
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A novel coupled-line structure is proposed to design dual-band and high-power Gysel power dividers with inherent impedance-transforming functions. Based on traditional even- and odd-mode technique, the analytical design methods in closed-form formula are obtained and the accurate electrical parameters analysis is presented. Due to the usage of coupled-line sections, more design-parameter freedom and a wider frequency-ratio operation range for this kind of dual-band Gysel powder divider are obtained. Several numerical examples are designed and calculated to demonstrate flexible dual-band applications with different impedance-transforming functions. A practical microstrip power divider operating at 2 GHz and 3.2 GHz is designed, fabricated, and measured. The good agreement between the calculated and measured results verifies our proposed circuit structure and analytical design approach.
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Comparative analysis of mitochondrial genomes in distinct nuclear ploidy loach Misgurnus anguillicaudatus and its implications for polyploidy evolution.
PLoS ONE
PUBLISHED: 01-01-2014
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Misgurnus anguillicaudatus has several natural ploidy types. To investigate whether nuclear polyploidy have an impact on mitochondrial DNA (mtDNA), the complete mitochondrial genomes (mitogenomes) of five distinct ploidy M. anguillicaudatus (natural diploid, triploid, tetraploid, pentaploid and hexaploid), which were collected in central China, were sequenced and analyzed. The five mitogenomes share the same gene arrangement and have similar gene size, base composition and codon usage pattern. The most variable regions of the mitogenome were the protein-coding genes, especially the ND4L (5.39% mutation rate). Most variations occurred in tetraploids. The phylogenetic tree showed that the tetraploid M. anguillicaudatus separated early from other ploidy loaches. Meanwhile, the mitogenomes from pentaploids, and hexaploids have the closest phylogenetic relations, but far from that of tetraploids, implying that pentaploids and hexaploids could not be formed from tetraploids, possibly from the diploids and triploids. The genus Misgurnus species were divided into two divergent inter-genus clades, and the five ploidy M. anguillicaudatus were monophyletic, which support the hypotheses about the mitochondrial introgression in loach species.
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Intensive insulin therapy increases sex hormone-binding globulin in newly diagnosed type 2 diabetic patients.
Eur. J. Endocrinol.
PUBLISHED: 01-01-2014
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Many studies have shown that low sex hormone-binding globulin (SHBG) is associated with insulin resistance, but only few studies have examined how serum SHBG is regulated by insulin in humans. This interventional study aimed to investigate the effect of insulin therapy (IT) on serum SHBG levels in newly diagnosed type 2 diabetic patients.
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HTLV-1 Tax-deregulated both autophagy pathway and c-FLIP expression contribute to the resistance against death receptor-mediated apoptosis.
J. Virol.
PUBLISHED: 12-18-2013
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The human T-cell leukemia virus type 1 Tax protein is considered to play a central role in the process that leads to adult T-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). HTLV-1 Tax-expressing cells show resistance to apoptosis induced by Fas ligand (FasL) and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL). The regulation of Tax on autophagy pathway is recently reported in HeLa cells and peripheral T cells, but the function and underlying molecular mechanism of the Tax-regulated autophagy are not yet well defined. Here, we report that HTLV-1 Tax deregulates autophagy pathway, which plays a protective role during the death receptor (DR)-mediated apoptosis of human U251 astroglioma cells. The cellular FLICE-inhibitory protein (c-FLIP), which is upregulated by Tax, also contributes to the resistance against DR-mediated apoptosis. Both Tax-induced autophagy and Tax-induced c-FLIP expression require Tax-induced activation of I?B kinases (IKK). Furthermore, Tax-induced c-FLIP expression is regulated through the Tax-IKK-NF-?B signaling pathway, whereas Tax-triggered autophagy depends on the activation of IKK, but not the activation of NF-?B. In addition, DR-mediated apoptosis is correlated with the degradation of Tax, which can be facilitated by the inhibitors of autophagy.
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The complete mitochondrial genome of natural Misgurnus bipartitus (Cypriniformes: Cobitidae).
Mitochondrial DNA
PUBLISHED: 10-15-2013
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Abstract Misgurnus bipartitus, a small sized freshwater fish species, is one of the most important cultured fish in East Asia. In this study, the complete mitochondrial genome of M. bipartitus is sequenced to be 16,636?bp in length, including 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, a control region and the origin of the light strand replication. The overall base composition of M. bipartitus in descending order is A 29.79 %, T 27.95%, C 25.90% and G 16.36%, with a slight A?+?T bias. The mitogenome sequence data may provide useful information to the population genetics analysis of M. bipartitus and the elucidation of evolutionary mechanisms in Cobitidae.
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Molecular cloning and expression analysis of mannose receptor C type 1 in grass carp (Ctenopharyngodon idella).
Dev. Comp. Immunol.
PUBLISHED: 09-17-2013
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Mannose receptor C type 1 (MRC1) is a pattern-recognition receptor (PRR) which plays a significant role in immune responses. Much work on MRC1 has been done in mammals and birds while little in fish. In this study, we cloned and characterized MRC1 in grass carp (gcMR). The full-length gcMR contained 5291bp encoding a putative protein of 1432 amino acids. The predicted amino acid sequences showed that gcMR contained a signal peptide, a cysteine-rich (CR) domain, a fibronectin type II (FN II) domain, eight C-type lectin-like domains (CTLDs), a transmembrane domain and a short cytoplasmic domain. gcMR were constitutively expressed in different organs with the higher expression in spleen and head kidney. During embryonic development, gcMR transcript levels were highest at cleavage stage. The up-regulation expression of gcMR, IL-1? and TNF-? in liver, spleen, head kidney and intestine after Aeromonas hydrophila infection indicating it involved in innate immune regulation during bacterial infections.
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The development of an in vitro assay to screen lipid based nanoparticles for siRNA delivery.
J Control Release
PUBLISHED: 08-28-2013
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In order to rapidly screen and select lead candidates for in vivo evaluation of lipid nanoparticles (LNPs) for systemic small interfering RNA (siRNA) delivery, an in vitro assay amenable to high-throughput screening (HTS) is developed. The strategy is to mimic the in vivo experience of LNPs after systemic administration, such as interactions with serum components, exposure to endosomal pH environments, and interactions with endosomal membrane lipids. It is postulated that the amount of siRNA released from LNPs after going through these treatments can be used as a screening tool to rank order the effectiveness of siRNA delivery by lipid nanoparticles in vivo. LNPs were incubated with 50% serum from different species (i.e. mouse, rat, or rhesus) at 37°C. The resulting samples were then reacted with anionic, endosomal-mimicking lipids at different pHs. The amount of siRNA released from LNPs was determined using spectrophotometry employing the fluorescent indicator SYBR Gold. Our results indicated that the amount of siRNA liberated was highly dependent upon the species of serum used and the pH to which it was exposed. LNPs treated with mouse serum showed higher levels of siRNA release, as did those subjected to endosomal pH (6.0), compared to physiological pH. Most interestingly, a good correlation between the amount of siRNA released and the in vivo efficacy was observed. In conclusion, an in vitro siRNA release assay was developed to screen and rank order LNPs for in vivo evaluation.
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The opposite prognostic significance of nuclear and cytoplasmic p21 expression in resectable gastric cancer patients.
J. Gastroenterol.
PUBLISHED: 08-01-2013
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Protein p21(Cip1/Waf1) is a cyclin-dependent kinase inhibitor, which plays important roles in cell cycle arrest, senescence, and apoptosis. Interestingly, the nuclear and cytoplasmic p21 executes various functions in the cell. In this study, we investigated the prognostic impact of subcellular p21 expression in gastric cancer (GC).
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One-Year Outcomes from an All-Comers Chinese Population of Patients Implanted With the Resolute Zotarolimus-Eluting Stent.
Am. J. Cardiol.
PUBLISHED: 07-31-2013
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The RESOLUTE China Registry is a prospective, multicenter, all-comers, observational study of patients in China implanted with the Resolute zotarolimus-eluting stent (R-ZES). R-ZES was commercially available before the enrollment began. All patients suitable for R-ZES implantation according to applicable guidelines were candidates for enrollment at 30 centers and were treated per standard hospital practice. Dual antiplatelet therapy (DAPT) was prescribed for a minimum of 6 months per current European Society of Cardiology guidelines and the device instructions for use. There were 1,800 patients enrolled with a mean age of 61.3 ± 10.9 years, 76% of patients were men, and 61% had complex disease. DAPT use was 94% at 1 year. Target lesion failure (cardiac death, target vessel myocardial infarction, or clinically driven target lesion revascularization) at 1 year was 3.5% (95% confidence interval 2.7% to 4.5%). The rate of cardiac death was 0.6%, target vessel myocardial infarction 2.3%, and clinically driven target lesion revascularization 0.9%. The 1-year rate of definite or probable stent thrombosis was 0.5% (8 of 1,750); 0.4% (7 of 1,750) occurred early (0 to 30 days) and 1 event occurred late (1 to 12 months). One stent thrombosis occurred in a patient who had an interruption of DAPT within the first month; all other stent thromboses occurred while on DAPT. Outcomes did not differ significantly between monitored and unmonitored patients (difference in target lesion failure, p = 0.264). In conclusion, the RESOLUTE China Registry confirms the safety and effectiveness of R-ZES in a large real-world Chinese population.
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The complete nucleotide sequence of white Amur bream (Parabramis pekinensis) mitochondrial genome.
Mitochondrial DNA
PUBLISHED: 07-31-2013
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Abstract White Amur bream, Parabramis pekinensis (Cypriniformes: Cyprinidae), a freshwater cyprinid fish, is an important economic fish in several countries, especially in China. The complete sequence of P. pekinensis mitochondrial genome has been determined. The genome is 16,622?bp in length, and consists of 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and the noncoding control region, with the genomic organization being identical to that of typical vertebrates. Three conserved sequence blocks (CSB1 to CSB3) were identified in the control region. The complete mitochondrial genome sequence is useful for phylogenetic analysis and studies of population genetics of P. pekinensis.
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14-3-3? regulates the proliferation of glioma cells through the GSK3?/?-catenin signaling pathway.
Oncol. Rep.
PUBLISHED: 07-19-2013
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We previously demonstrated that 14-3-3? is overexpressed in astrocytomas; however, the underlying mechanisms are poorly understood. Based on the reported multiple functions of 14-3-3?, we hypothesized that it interacts with glycogen synthase kinase 3 ? (GSK3?), which regulates ?-catenin-mediated oncogene expression and contributes to tumorigenesis and astrocytoma progression. To test these hypotheses, we used 14-3-3? overexpression vectors and small interfering RNA (siRNA) transfection in the human normal astrocyte cell line SVGp12 and the glioma cell line U87, respectively. The results showed that overexpression of 14-3-3? promoted the proliferation of SVGp12 cells, while knockdown of 14-3-3? inhibited the proliferation of U87 cells as analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and bromodeoxyuridine (BrdU) assays. In Flag-tagged 14-3-3?-overexpressing cells, GSK3? was co-immunoprecipitated with 14-3-3? using a Flag antibody. Knockdown of ?-catenin by siRNA blocked cell proliferation induced by overexpression of 14-3-3?. Furthermore, overexpression of 14-3-3? suppressed the phosphorylation of ?-catenin leading to its accumulation and nuclear translocation as revealed by western blot analysis. In addition, ?-catenin nuclear translocation induced by overexpression of 14-3-3? activated the transcription of oncogenes including c-myc and cyclin D1. Collectively, these results revealed that 14-3-3? regulates the proliferation of astrocytes and glioma cells through the GSK3?/?-catenin signaling pathway. The delineated mechanism of 14-3-3? may be responsible for the tumorigenesis and progression of human astrocytomas. Thus, new therapeutic strategies or drugs aimed at 14-3-3? may have potential for the treatment of human astrocytomas.
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Generation of diffraction-free optical beams using wrinkled membranes.
Sci Rep
PUBLISHED: 07-09-2013
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Wrinkling has become a well developed bottom-up technique to make artificial surface textures in about the last decade. However, application of the optical properties of long range ordered wrinkles has been limited to one dimensional gratings to date. We report the demonstration of macroscopic optical focusing using wrinkled membranes, in which concentric wrinkle rings on a gold-PDMS bilayer membrane convert collimated illuminations to diffraction-free focused beams. Beam diameters of 300-400??m have been observed in the visible range, which are dominantly limited by the eccentricity of the current devices. Based upon agreement between theoretical and experimental results on eccentricity effects, we predict a decrease of the beam diameter to no more than around 50??m, if eccentricity is eliminated.
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The complete mitochondrial genomes of natural diploid and tetraploid loaches Misgurnus anguillicaudatus (Cypriniformes: Cobitidae).
Mitochondrial DNA
PUBLISHED: 06-24-2013
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Abstract Dojo loach Misgurnus anguillicaudatus, a small-sized freshwater fish species, is not only one of the most important cultured fish in East Asia, but also a promising model animal to study evolutionary biology for polyploidy. We sequenced the complete mitochondrial genomes from the natural diploid and tetraploid individuals of this species. The two different cytotypes loaches share the same length (16,646?bp) and structural organization of mitochondrial genome. The two genomes have 98.3% nucleotide sequence similarity, and the biggest nucleotide sequence divergence between homologous genes is observed in CYTB gene.
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Low serum sex hormone-binding globulin is associated with nonalcoholic fatty liver disease in type 2 diabetic patients.
Clin. Endocrinol. (Oxf)
PUBLISHED: 06-16-2013
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Studies have indicated that low serum sex hormone-binding globulin (SHBG) and testosterone levels are associated with nonalcoholic fatty liver disease (NAFLD). However, it remains unclear whether an association exists between SHBG and NAFLD independent of testosterone.
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Expression of heat shock protein 90 genes during early development and infection in Megalobrama amblycephala and evidence for adaptive evolution in teleost.
Dev. Comp. Immunol.
PUBLISHED: 05-03-2013
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Heat shock protein 90 (HSP90) are highly conserved molecular chaperones, playing a pivotal role in cellular progress. In this study, we reported the characterization of the Hsp90? and Hsp90? genes in Megalobrama amblycephala, the expression profiling during early development, in various healthy tissues and in response to bacterial infection, and the assessment of their adaptive evolution. The Hsp90? cDNA contains an open reading frame (ORF) of 2193 bp encoding 731 amino acids and the Hsp90? cDNA has an ORF of 2184 bp encoding 728 amino acids. Using quantitative real-time PCR (qRT-PCR) analysis, the mRNA of both Hsp90? and Hsp90? reached the highest level at 15th day post-hatch. Using qRT-PCR and Western blot, both Hsp90? and Hsp90? were widely expressed in various healthy tissues and significantly higher in blood than in other tissues. Expression of both Hsp90? and Hsp90? were up-regulated upon bacterial infection and reached the peak level at 4 h post infection. Site model analysis indicated that one positive selection site (T717) in Hsp90? was found, while no positive selection site was observed in Hsp90?. Branch-site model test showed that there were adaptively evolutionary evidences in the branches of Salmoniformes and Gasterosteiformes for Hsp90? gene, and in the branch of Salmoniformes for Hsp90? gene.
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Anti-stress properties and two HSP70s mRNA expressions of blunt snout bream (Megalobrama amblycephala) fed with all-plant-based diet.
Fish Physiol. Biochem.
PUBLISHED: 04-23-2013
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The influence of all-plant-based diet on fingerling blunt snout breams (Megalobrama amblycephala) was tested by examining growth performance, anti-stress properties and related gene expression. Healthy fish were randomly divided into triplicate groups per dietary treatment and fed with different formulated diets. The results showed that both weight gain, specific growth rate and protein efficiency ratio of all-plant-based diet group were significant higher than those of the control (p < 0.05). In contrast, FCR of all-plant-based diet group was significantly lower than that of the control (p < 0.05). Therefore, all-plant-based diets could not affect the growth performance of blunt snout breams. Compared to the control group, the lysozyme levels in serum and mucus, and glutamic-oxaloacetic transaminase activities in serum and liver decreased significantly (p < 0.05). In contrast, the glutamic-pyruvic transaminase activities in serum and liver increased significantly (p < 0.05). For blunt snout breams fed with all-plant-based diets, the superoxide dismutase activities in mucus, serum and liver as well as catalase activity in serum and liver were decreased significantly (p < 0.05) comparing with that of the control group. But malondialdehyde contents were higher (p < 0.05) in serum and liver than that of control group. The expression of HSC70 mRNA increased significantly (p < 0.05) in blunt snout breams fed with all-plant-based diet, whereas the HSP70 mRNA expression decreased significantly (p < 0.05) when compared with control group. In conclusion, all these results indicated that the application of all-plant-based diet could decrease the anti-stress properties (non-specific immunity, stress resistance and antioxidant ability) and HSP70 mRNA expression in blunt snout breams fingerling. Although all-plant-based diets could not affect the growth performance of blunt snout breams, the application of all-plant-based diet should be discreet in the production practice.
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Photoreceptors in whirler mice show defective transducin translocation and are susceptible to short-term light/dark changes-induced degeneration.
Exp. Eye Res.
PUBLISHED: 04-11-2013
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Usher syndrome combines congenital hearing loss and retinitis pigmentosa (RP). Mutations in the whirlin gene (DFNB31/WHRN) cause a subtype of Usher syndrome (USH2D). Whirler mice have a defective whirlin gene. They have inner ear defects but usually do not develop retinal degeneration. Here we report that, in whirler mouse photoreceptors, the light-activated rod transducin translocation is delayed and its activation threshold is shifted to a higher level. Rhodopsin mis-localization is observed in rod inner segments. Continuous moderate light exposure can induce significant rod photoreceptor degeneration. Whirler mice reared under a 1500 lux light/dark cycle also develop severe photoreceptor degeneration. Previously, we have reported that shaker1 mice, a USH1B model, show moderate light-induced photoreceptor degeneration with delayed transducin translocation. Here, we further show that, in both whirler and shaker1 mice, short-term moderate light/dark changes can induce rod degeneration as severe as that induced by continuous light exposure. The results from shaker1 and whirler mice suggest that defective transducin translocation may be functionally related to light-induced degeneration, and these two symptoms may be caused by defects in Usher protein function in rods. Furthermore, these results indicate that both Usher syndrome mouse models possess a light-induced retinal phenotype and may share a closely related pathobiological mechanism.
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Endosomolytic bioreducible poly(amido amine disulfide) polymer conjugates for the in vivo systemic delivery of siRNA therapeutics.
Bioconjug. Chem.
PUBLISHED: 03-25-2013
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Efficient siRNA delivery is dependent not only on the ability of the delivery vehicle to target a specific organ but also on its ability to enable siRNA entry into the cytoplasm of the target cells. Polymers with endosomolytic properties are increasingly being used as siRNA delivery vehicles due to their potential to facilitate endosomal escape and intracellular delivery. Addition of disulfide bonds in the backbone of these polymers was expected to provide degradability through reduction by glutathione in cytosol. This paper describes the synthesis of new endosomolytic bioreducible poly(amido amine disulfide) polymers whose lytic potential can be masked at physiological pH, but can be restored at acidic endosomal pH. These polymer conjugates gave good in vitro knockdown (KD) and did not demonstrate cytotoxicity in a MTS assay. Efficient mRNA KD for apolipoprotein B in mouse liver was observed with these polyconjugates following intravenous dosing.
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Proteomic analysis of epithelioma papulosum cyprini cells infected with spring viremia of carp virus.
Fish Shellfish Immunol.
PUBLISHED: 03-20-2013
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Spring viremia of carp (SVC), caused by spring viremia of carp virus (SVCV) is an important disease due to its drastic effects on carp fisheries in many countries. To better understand molecular responses to SVCV infection, two dimensional electrophoresis (2-DE) and MALDI-TOF/TOF were performed to investigate altered proteins in epithelioma papulosum cyprini cells (EPCs). Differentially expressed proteins in mock-infected EPCs and SVCV-infected EPCs were compared. A total of 54 differentially expressed spots were successfully identified (33 up-regulated spots and 21 down-regulated spots) which include cytoskeleton proteins, macromolecular biosynthesis-associated proteins, stress response proteins, signal transduction proteins, energy metabolism, and ubiquitin proteasome pathway-associated proteins. Moreover, 7 corresponding genes of the differentially expressed proteins were quantified using real time RT-PCR to examine their transcriptional profiles. The presence of four selected cellular proteins (beta-actin, gamma1-actin, heat shock cognate 71 kDa protein and annexin A2) associated with the spring viremia of carp virus (SVCV) particles was validated by Western blot assay. This study provides dynamic and useful protein-related information to further understand the underlying pathogenesis of SVCV infection.
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RASSF10 is epigenetically silenced and functions as a tumor suppressor in gastric cancer.
Biochem. Biophys. Res. Commun.
PUBLISHED: 02-03-2013
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Ras association domain family (RASSF) proteins are encoded by several tumor suppressor genes that are frequently silenced in human cancers. In this study, we investigated RASSF10 as a target of epigenetic inactivation and examined its functions as a tumor suppressor in gastric cancer. RASSF10 was silenced in six out of eight gastric cancer cell lines. Loss or downregulation of RASSF10 expression was associated with promoter hypermethylation, and could be restored by a demethylating agent. Overexpression of RASSF10 in gastric cancer cell lines (JRST, BGC823) suppressed cell growth and colony formation, and induced apoptosis, whereas RASSF10 depletion promoted cell growth. In xenograft animal experiments, RASSF10 overexpression effectively repressed tumor growth. Mechanistic investigations revealed that RASSF10 inhibited tumor growth by blocking activation of ?-catenin and its downstream targets including c-Myc, cyclinD1, cyclinE1, peroxisome proliferator-activated receptor ?, transcription factor 4, transcription factor 1 and CD44. In conclusion, the results of this study provide insight into the role of RASSF10 as a novel functional tumor suppressor in gastric cancer through inhibition of the Wnt/?-catenin signaling pathway.
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New cytochrome P450 1B1, 1C1, 2Aa, 2Y3, and 2K genes from Chinese rare minnow (Gobiocypris rarus): Molecular characterization, basal expression and response of rare minnow CYP1s and CYP2s mRNA exposed to the AHR agonist benzo[a]pyrene.
Chemosphere
PUBLISHED: 01-30-2013
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Cytochrome P450 (CYP450) genes play an important role in catalyzing oxidative metabolism of toxicants. Recently, CYP1 subfamily were discovered and reported in fish, however, little is known regarding the CYP2 isoforms in fish. In the present study, the cDNA fragments of CYP 1B1 and 1C1 and CYP2Aa, 2Y3, and 2K of rare minnow were cloned and exhibited a high amino acid sequence identity compared with their zebrafish orthologs. Basal expression showed CYP1C1 and CYP 2Aa expression were observed in all eight tissues analyzed (liver, gill, intestine, kidney, spleen, brain, skin, and muscle). CYP 1A, and 1B1 expression was found in all tissues except for muscle and skin. However, CYP 2Y3 was expressed in liver, spleen, intestine and muscle whereas CYP 2K in liver, kidney and intestine. 4 and 100?gL(-1) Benzo[a]pyrene (BaP) induced patterns showed that CYP 1A, 1B1 and 1C1 expression in liver, gill, and intestine was strongly up-regulated (p<0.05). Furthermore, CYP 2Y3 was strongly induced in liver from BaP treatments (p<0.05). The high induction on mRNA level of CYP1s and CYP 2Y3 by BaP could be associated with catalyzing detoxification and indicated that CYP2s may also be potential biomarker to screen AHR agonist. The high responsiveness of CYP1 and 2 genes suggested Chinese rare minnow is feasible to screen and assess pollution with AHR agonist.
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Identification of HIF-1? promoter and expression regulation of HIF-1? gene by LPS and hypoxia in zebrafish.
Fish Physiol. Biochem.
PUBLISHED: 01-22-2013
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The ubiquitously expressed hypoxia-inducible factor-1? (HIF-1?) acts as a key transcription factor in regulating metabolism, development, cellular survival, proliferation and pathology under hypoxia condition. Compared to mammals, fish are more vulnerable to hypoxia stress and contamination; however, the regulation of HIF-1? in fish remains obscure. In this study, zebrafish HIF-1? promoter was cloned and found to possess a CpG island located at -97 to +403, but the canonical TATA-box was absent. Aligning 240-bp HIF-1? proximal promoter region of zebrafish with other vertebrates showed more than 82 % identity with cyprinid fishes. Further luciferase analysis suggested that the minimal core promoter might locate at -134 to +97, and several putative transcription factor binding sites were found in this region by bioinformatic analysis. Moreover, it was shown that the zebrafish HIF-1? mRNA was significantly activated by 10 ?g/mL lipopolysaccharide (LPS) under hypoxia condition and peaked at 8 h after treatment, suggesting LPS- and hypoxia-regulated zebrafish HIF-1? transcriptional activity in a synergistic pattern. This synergistic effect was closely related to the living environment of fish, indicating that this mechanism would be more conducive to fish survival.
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Antioxidant response of soybean seedlings to joint stress of lanthanum and acid rain.
Environ Sci Pollut Res Int
PUBLISHED: 01-08-2013
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Excess of rare earth elements in soil can be a serious environmental stress on plants, in particular when acid rain coexists. To understand how such a stress affects plants, we studied antioxidant response of soybean leaves and roots exposed to lanthanum (0.06, 0.18, and 0.85 mmol L(-1)) under acid rain conditions (pH 4.5 and 3.0). We found that low concentration of La3+ (0.06 mmol L(-1)) did not affect the activity of antioxidant enzymes (catalase and peroxidase) whereas high concentration of La3+ (?0.18 mmol L(-1)) did. Compared to treatment with acid rain (pH 4.5 and pH 3.0) or La3+ alone, joint stress of La3+ and acid rain affected more severely the activity of catalase and peroxidase, and induced more H2O2 accumulation and lipid peroxidation. When treated with high level of La3+ (0.85 mmol L(-1)) alone or with acid rain (pH 4.5 and 3.0), roots were more affected than leaves regarding the inhibition of antioxidant enzymes, physiological function, and growth. The severity of oxidative damage and inhibition of growth caused by the joint stress associated positively with La3+ concentration and soil acidity. These results will help us understand plant response to joint stress, recognize the adverse environmental impact of rare earth elements in acidic soil, and develop measures to eliminate damage caused by such joint stress.
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Light-induced translocation of RGS9-1 and G?5L in mouse rod photoreceptors.
PLoS ONE
PUBLISHED: 01-04-2013
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The transducin GTPase-accelerating protein complex, which determines the photoresponse duration of photoreceptors, is composed of RGS9-1, G?5L and R9AP. Here we report that RGS9-1 and G?5L change their distribution in rods during light/dark adaptation. Upon prolonged dark adaptation, RGS9-1 and G?5L are primarily located in rod inner segments. But very dim-light exposure quickly translocates them to the outer segments. In contrast, their anchor protein R9AP remains in the outer segment at all times. In the dark, G?5Ls interaction with R9AP decreases significantly and RGS9-1 is phosphorylated at S(475) to a significant degree. Dim light exposure leads to quick de-phosphorylation of RGS9-1. Furthermore, after prolonged dark adaptation, RGS9-1 and transducin G? are located in different cellular compartments. These results suggest a previously unappreciated mechanism by which prolonged dark adaptation leads to increased light sensitivity in rods by dissociating RGS9-1 from R9AP and redistributing it to rod inner segments.
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Overexpression of arginine transporter CAT-1 is associated with accumulation of L-arginine and cell growth in human colorectal cancer tissue.
PLoS ONE
PUBLISHED: 01-01-2013
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We previously showed that L-arginine (Arg) accumulates in colorectal cancer tissues. The aim of this study was to investigate the mechanism by which Arg accumulates and determine its biological significance. The concentration of Arg and Citrulline (Cit) in sera and tumor tissues from colorectal cancer (CRC) patients was analyzed by high-performance liquid chromatography (HPLC). The expression of Arg transporters was analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemical analysis of tissue microarray. We also transfected the colon cancer cell line HCT-116 with siRNA specific for the Arg transporter CAT-1 and measured the induction of apoptosis by flow cytometry and cell proliferation by MTT assay. Consistent with our previous results, serum Arg and Cit concentrations in colorectal cancer patients were significantly lower than those in normal volunteers, while Arg and Cit concentrations in colorectal cancer tissues were significantly higher than in matched adjacent normal colon tissues. Quantitative RT-PCR showed that the CAT-1 gene was highly overexpressed in 70.5% of colorectal cancer tissue samples relative to adjacent normal colon tissues in all 122 patients with colorectal cancer. Immunohistochemical analysis of tissue microarray confirmed that the expression of CAT-1 was higher in all 25 colorectal cancer tissues tested. CAT-1 siRNA significantly induced apoptosis of HCT-116 cells and subsequently inhibited cell growth by 20-50%. Our findings indicate that accumulation of L-Arg and Cit and cell growth in colorectal cancer tissues is associated with over-expression of the Arg transporter gene CAT-1. Our results may be useful for the development of molecular diagnostic tools and targeted therapy for colorectal cancer.
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Generation and characterization of monoclonal antibodies against channel catfish virus.
Hybridoma (Larchmt)
PUBLISHED: 12-14-2011
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Three monoclonal antibodies (MAbs) against channel catfish virus (CCV) were generated from mice immunized with purified CCV. Western blot analysis revealed that the MAb 3G12 reacted with three CCV proteins of 94 kDa, 130 kDa, and 170 kDa; the MAb 4C4 reacted with two CCV proteins of 130 kDa and 170 kDa; and the MAb 4D4 reacted with two CCV proteins of 94 kDa and 98 kDa. Indirect immunofluorescence assay showed intense fluorescence in the CCV-infected channel catfish ovary (CCO) cells in areas corresponding to the location of granular structures. In addition, the three MAbs could completely neutralize CCV at a dilution of 1:500. This study demonstrated that these MAbs could recognize CCV specifically and will be useful in the development of diagnostic methods for the detection of fish CCV infection.
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Ethanol vapor sensing properties of triangular silver nanostructures based on localized surface plasmon resonance.
Sensors (Basel)
PUBLISHED: 07-25-2011
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A sensitive volatile organic vapor sensor based on the LSPR properties of silver triangular nanoprisms is proposed in this paper. The triangular nanoprisms were fabricated by a nanosphere lithography (NSL) method. They have sharp vertices and edges, and are arranged in an ideal hexangular array. These characteristics ensure that they exhibit an excellent LSPR spectrum and a high sensitivity to the exterior environment changes. The LSPR spectra responding to ethanol vapor and four other volatile organic vapors-acetone, benzene, hexane and propanol-were measured with a UV-vis spectrometer in real time. Compared with the other four vapors, ethanol exhibits the highest sensitivity (?0.1 nm/mg L(-1)) and the lowest detection limit (?10 mg/L) in the spectral tests. The ethanol vapor test process is also fast (?4 s) and reversible. These insights demonstrate that the triangular nanoprism based nano-sensor can be used in ethanol vapor detection applications.
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DNA extraction from crayfish exoskeleton.
Indian J. Exp. Biol.
PUBLISHED: 07-07-2011
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Crayfish exoskeleton (CE) samples are generally less invasive and easy to be collected. However, it is difficult to extract DNA from them. This study was intended to investigate CE as a DNA source and design an easy and efficient DNA extraction protocol for polymerase chain reactions. Specific primer pair (PPO-F, PPO-R) was used to amplify extracted DNA from CE, and compared to crayfish tail muscle DNA sample. Moreover, seven microsatellites markers were used to amplify the CE DNA samples set. Since the extracted DNA from CE is suitable for gene amplification, the results present usefulness of CE as an easy and convenient DNA source for PCR-based population genetic research.
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Syk protein tyrosine kinase involves PECAM-1 signaling through tandem immunotyrosine inhibitory motifs in human THP-1 macrophages.
Cell. Immunol.
PUBLISHED: 05-25-2011
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Although recent evidence supports a functional relationship between platelet endothelial cell adhesion molecule (PECAM-1) and Syk tyrosine kinase, little is known about the interaction of Syk with PECAM-1. We report that down-regulation of Syk inhibits the spreading of human THP-1 macrophage cells. Moreover, our data indicate that Syk binds PECAM-1 through its immune tyrosine-based inhibitory motif (ITIM), and dual phosphorylation of the ITIM domain of PECAM-1 leads to activation of Syk. Our results indicate that the distance between the phosphotyrosines could be up to 22 amino acids in length, depending on the conformational flexibility, and that the dual ITIM tyrosine motifs of PECAM-1 facilitate immunoreceptor tyrosine-based activation motif-like signaling. The preferential binding of PECAM-1 to Src homology region 2 domain-containing phosphatase-2 or Syk may depend on their relative affinities, and could provide a mechanism by which signal transduction from PECAM-1 is internally regulated by both positive and negative signaling enzymes.
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Deletion of Serpina1a, a murine ?1-antitrypsin ortholog, results in embryonic lethality.
Exp. Lung Res.
PUBLISHED: 05-16-2011
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Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in the United States Approximately 1% to 2% of COPD patients suffer from ?(1)-antitrypsin (A1AT) deficiency, the major inheritable predisposition to COPD/emphysema. To further study the role of A1AT deficiency in the pathogenesis of COPD/emphysema, the authors attempted to generate null-mutant mice for Serpina1a, 1 of 2 A1AT orthologs in mice. Here the authors show that targeted deletion of Serpina1a results in embryonic lethality prior to 8.5 days post conception (dpc). The results are surprising given that A1AT-null humans exist and therefore do not require this gene product for normal development. The Serpina1 gene cluster is substantially different between mouse and man. Through gene duplication, mice have 3 to 5 (depending on the strain) highly homologous proteinase inhibiting (Pi) genes, 2 of which inhibit neutrophil elastase. Despite the abundance of Pi genes in mice, Serpina1a serves a critical, nonredundant function during early mouse development. A1AT-deficient mice have been highly sought after to study emphysema, cancer, and liver disease, and as a model to perfect gene replacement therapy. These results highlight important differences between human and murine serpins and point to the difficulty inherent to using gene-targeted mice to study this common human genetic disease.
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Isolation of New 40 Microsatellite Markers in Mandarin Fish (Siniperca chuatsi).
Int J Mol Sci
PUBLISHED: 05-05-2011
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In this study, 23 genomic microsatellite DNA markers and 17 express sequence tag (EST)-derived microsatellites were developed and characterized using the fast isolation by AFLP of sequences containing repeats (FIASCO) method and data mining from public EST databases of mandarin fish (Siniperca chuatsi). These polymorphic microsatellite markers were then tested for polymorphism in a wild S. chuatsi population. The number of alleles at 23 genomic SSRs varied from 2 to 19 with an average of 8.0 alleles per locus. The average observed and expected heterozygosities were 0.746 and 0.711, respectively. Of 5361 EST sequences examined, 3.9% (209) contain microsatellites, and di-nucleotide repeats are the most abundant (67.0%), followed by tri-nucleotide (29.7%) and tetra-nucleotide repeats (3.3%). The number of alleles at 17 EST-SSRs varied from 2 to 17 with an average of 8.4 alleles per locus. The average observed and expected heterozygosities were 0.789 and 0.685, respectively. No significant difference of loci polymorphism was found between genomic SSRs and EST-SSRs in terms of number of alleles and heterozygosities. Results of cross-species utility indicated that 13 (52.2%) of the genomic-SSRs and 13 (76.5%) of the EST-SSRs were successfully cross-amplified in a related species, the golden mandarin fish (Siniperca scherzeri).
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Molecular test for vivax malaria with loop-mediated isothermal amplification method in central China.
Parasitol. Res.
PUBLISHED: 04-13-2011
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Vivax malaria has the highest incidence in central China. High-throughput and cost-effective testing methods are essential for vivax malaria diagnosis in this area. Loop-mediated isothermal amplification (LAMP) is an established nucleic acid amplification method, which provides a promising platform for the molecular detection of malaria parasites in development countries. This study was performed to compare the LAMP method, the nested PCR-based method, and microscopic examinations in diagnosing vivax malaria. LAMP detected vivax malaria parasites in 160 of 164 microscopically positive samples (sensitivity, 97.6%), whereas nested PCR detected Plasmodium vivax in 162 of 164 samples (sensitivity, 98.8%). No false-positive results were obtained by LAMP or nested PCR among fever-positive and healthy samples. The sensitivities and specificities of the two molecular tests were consistently high. In addition, the reproducibility of the LAMP assays using water bath, which reduced the cost substantially. LAMP method is an accurate, rapid, simple, and cost-effective method that may be useful for diagnosis in field diagnoses instead of nested PCR. This method is feasible to diagnose vivax malaria parasite in endemic areas of central China.
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CXC chemokine ligand 16 as a prognostic marker in patients with intermediate coronary artery lesions: a 2-year follow-up study.
Tohoku J. Exp. Med.
PUBLISHED: 04-07-2011
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There is no reliable way to identify the high-risk patients with intermediate coronary artery lesions (diameter stenosis 20%-70%) in early stage. Soluble CXC chemokine ligand 16 (CXCL16) is a newly discovered chemokine that can mediate inflammatory responses. It is released by proteolytic cleavage of its membrane-bound form, named scavenger receptor for phosphatidylserine and oxidized lipoprotein (SR-PSOX) that can promote the uptake of oxidized low-density lipoprotein cholesterol by macrophages. We have hypothesized that CXCL16 is an indicator of the prognosis of intermediate coronary artery lesions, and thus assessed the association between plasma CXCL16 concentrations and the 2-year prognosis in 616 patients with intermediate coronary artery lesions. The primary endpoint was a composite of all-cause death, non-fatal myocardial infarction, revascularization and angina pectoris requiring re-hospitalization. During the median follow-up time of 24 months, 69 events occurred. The plasma concentrations of CXCL16 (median 7712.88 pg/ml vs. 6792.43 pg/ml, P = 0.014) and high-sensitivity C-reactive protein (hs-CRP) (median 2.82 mg/L vs. 1.68 mg/L, P < 0.001) were higher in patients with events than patients without events. Cox hazard proportion analysis showed patients in upper CXCL16 quartile were more likely to suffer from adverse outcome than patients in lower quartile (RR = 1.271, P = 0.029, 95% CI: 1.025-1.577) after adjusting for sex, age, smoking, hypertension, diabetes, fat, dyslipidemia, hs-CRP, and medication use. In conclusion, plasma level of CXCL16 is an independent predictor of the prognosis of the patients with intermediate coronary lesions. Elevated plasma CXCL16 is associated with higher risk for these patients.
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Permanent genetic resources added to Molecular Ecology Resources Database 1 December 2010-31 January 2011.
Mol Ecol Resour
PUBLISHED: 04-01-2011
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This article documents the addition of 238 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Alytes dickhilleni, Arapaima gigas, Austropotamobius italicus, Blumeria graminis f. sp. tritici, Cobitis lutheri, Dendroctonus ponderosae, Glossina morsitans morsitans, Haplophilus subterraneus, Kirengeshoma palmata, Lysimachia japonica, Macrolophus pygmaeus, Microtus cabrerae, Mytilus galloprovincialis, Pallisentis (Neosentis) celatus, Pulmonaria officinalis, Salminus franciscanus, Thais chocolata and Zootoca vivipara. These loci were cross-tested on the following species: Acanthina monodon, Alytes cisternasii, Alytes maurus, Alytes muletensis, Alytes obstetricans almogavarii, Alytes obstetricans boscai, Alytes obstetricans obstetricans, Alytes obstetricans pertinax, Cambarellus montezumae, Cambarellus zempoalensis, Chorus giganteus, Cobitis tetralineata, Glossina fuscipes fuscipes, Glossina pallidipes, Lysimachia japonica var. japonica, Lysimachia japonica var. minutissima, Orconectes virilis, Pacifastacus leniusculus, Procambarus clarkii, Salminus brasiliensis and Salminus hilarii.
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Expression characterization and the promoter activity analysis of zebrafish hdac4.
Fish Physiol. Biochem.
PUBLISHED: 03-25-2011
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Histone deacetylase 4 (HDAC4) is an important modifier enzyme for chromatin remodeling and plays an essential role in regulating gene expression. Spatio-temporal expression spectrum revealed that zebrafish hdac4 mRNA, ubiquitously distributed in various tissues, were significantly higher at 36 hpf (hours post-fertilization) and 6 dpf (days post-fertilization) than other periods. Trichostatin A (TSA) inhibited the development of zebrafish embryos and transcription of hdac4 and mef2a (myocyte enhancer factor-2A). Moreover, five vectors containing different promoter regions of hdac4 were constructed in order to analyze promoter activity. The vector containing the region from -125 to +160 exhibited maximum luciferase activity that was approximately 30.3-fold and 58.9-fold higher than the control in two kinds of cells, respectively. By comparing the luciferase activities between the region from -302 to +30 and -698 to +30, it was suggested that the region between -698 and -302 might contain mild negative regulatory elements.
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Efficient gene delivery into fish cells by an improved recombinant baculovirus.
J. Virol. Methods
PUBLISHED: 02-14-2011
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Studies on transduction of mammalian cells have shown that baculovirus can be used as an effective vector for gene delivery. However, previous studies have found the gene delivery efficiency to be very low in differentiated fish cells. In this study, an improved recombinant baculovirus, containing cytomegalovirus immediate-early (CMV-IE) promoter and an enhanced green fluorescent protein (EGFP) gene as the reporter gene, was constructed. The transduction efficiency of recombinant baculovirus in several fish cell lines was measured by flow cytometry (FCM) and the persistence of EGFP was monitored by fluorescence microscopy. The results demonstrated that baculovirus can mediate the high efficiency of gene delivery into differentiated fish cells. Furthermore, it was found that growth medium is superior to PBS as the surrounding solution to enhance the transduction efficiency in some fish cells. In addition, transgene expression can persist for a lengthy period in fish cells, and attained highest level several days later after transduction. This study suggest that the improved recombinant baculovirus can be an excellent gene delivery vector for fish cells and also providing new insights into the transduction of vertebrate cells with baculovirus.
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Cloning, identification and accurate normalization expression analysis of PPAR? gene by GeNorm in Megalobrama amblycephala.
Fish Shellfish Immunol.
PUBLISHED: 02-13-2011
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Megalobrama amblycephala suffers from serious liver diseases recently and PPAR? gene has been reported to play an important role in the immune system of animal liver. On the basis of these facts, we have cloned and identified full-length cDNA of PPAR? and examined its expression patterns at different embryo developmental stages and in different tissues of adult and young fish in order to improve liver disease immunity of M. amblycephala. We also accurately normalized seven reference genes by GeNorm and calculated their gene expression normalization factors. The total length of PPAR? cDNA was 2021 bp, comprising of 214-bp 5-untranslated region; 1404-bp open reading frame (encoding 467-amino acids); and 403-bp 3-untranslated region. PPAR? peptide was predicted to consist of 4 domains, i.e. A/B, C, D, and E/F. PPAR? mRNAs were detected in different tissues of adult and young fish including adipose tissue, gill, heart, liver, spleen, kidney, white muscle, intestine, brain and gonad. In adult fish, the expression of PPAR? in white muscles was highest followed by liver and it was lowest in gonads. Its expression in male gonads was significantly higher than female gonads. In young fish, the expression of PPAR? was highest in brain, followed by intestines and it was lowest in spleen. At different embryo developmental stages, the expression of PPAR? was highest at 2 cells stage and it was lowest at gastrula stage, but it increased on first day after hatching. In unfertilized spermatozoa, the expression of PPAR? was higher than unfertilized ovum.
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Functionally significant nicotine acetylcholine receptor subunit ?5 promoter haplotypes are associated with susceptibility to lung cancer in Chinese.
Cancer
PUBLISHED: 01-18-2011
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Although recent genome-wide association studies have been conducted to reveal the relation between variations in the ?5 nicotinic receptor subunit and lung cancer in European and American populations, to the authors knowledge, no definite information on the role of nicotine acetylcholine receptor subunit ?5 (CHRNA5) in lung cancer risk has been obtained in a Chinese population.
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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.