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Find video protocols related to scientific articles indexed in Pubmed.
Molecular dynamics reveal a novel kinase-substrate interface that regulates protein translation.
J Mol Cell Biol
PUBLISHED: 11-19-2014
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A key control point in gene expression is the initiation of protein translation, with a universal stress response being constituted by inhibitory phosphorylation of the eukaryotic initiation factor 2? (eIF2?). In humans, four kinases sense diverse physiological stresses to regulate eIF2? to control cell differentiation, adaptation, and survival. Here we develop a computational molecular model of eIF2? and one of its kinases, the protein kinase R, to simulate the dynamics of their interaction. Predictions generated by coarse-grained dynamics simulations suggest a novel mode of action. Experimentation substantiates these predictions, identifying a previously unrecognized interface in the protein complex, which is constituted by dynamic residues in both eIF2? and its kinases that are crucial to regulate protein translation. These findings call for a reinterpretation of the current mechanism of action of the eIF2? kinases and demonstrate the value of conducting computational analysis to evaluate protein function.
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Regulation of synaptic development and function by the Drosophila PDZ protein Dyschronic.
Development
PUBLISHED: 10-30-2014
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Synaptic scaffold proteins control the localization of ion channels and receptors, and facilitate molecular associations between signaling components that modulate synaptic transmission and plasticity. Here, we define novel roles for a recently described scaffold protein, Dsychronic (DYSC), at the Drosophila larval neuromuscular junction. DYSC is the Drosophila homolog of whirlin/DFNB31, a PDZ domain protein linked to Usher syndrome, the most common form of human deaf-blindness. We show that DYSC is expressed presynaptically and is often localized adjacent to the active zone, the site of neurotransmitter release. Loss of DYSC results in marked alterations in synaptic morphology and cytoskeletal organization. Moreover, active zones are frequently enlarged and misshapen in dysc mutants. Electrophysiological analyses further demonstrate that dysc mutants exhibit substantial increases in both evoked and spontaneous synaptic transmission. We have previously shown that DYSC binds to and regulates the expression of the Slowpoke (SLO) BK potassium channel. Consistent with this, slo mutant larvae exhibit similar alterations in synapse morphology, active zone size and neurotransmission, and simultaneous loss of dysc and slo does not enhance these phenotypes, suggesting that dysc and slo act in a common genetic pathway to modulate synaptic development and output. Our data expand our understanding of the neuronal functions of DYSC and uncover non-canonical roles for the SLO potassium channel at Drosophila synapses.
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[Intensity loss of two-photon excitation fluorescence microscopy images of mouse oocyte chromosomes].
Guang Pu Xue Yu Guang Pu Fen Xi
PUBLISHED: 10-02-2014
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As an optical microscope with high resolution, two-photon excitation (TPE) fluorescence microscope is widely used in noninvasive 3D optical imaging of biological samples. Compared with confocal laser scanning microscope, TPE fluorescence microscope provides a deeper detecting depth. In spite of that, the image quality of sample always declines as the detecting depth increases when a noninvasive 3D optical imaging of thicker samples is performed. Mouse oocytes with a large diameter, which play an important role in clinical and biological fields, have obvious absorption and scattering effects. In the present paper, we performed compensation for two-photon fluorescence images of mouse oocyte chromosomes. Using volume as a parameter, the attenuation degree of these chromosomes was also studied. The result of our data suggested that there exists a severe axial intensity loss in two-photon microscopic images of mouse oocytes due to the absorption and scattering effects. It is necessary to make compensation for these images of mouse oocyte chromosomes obtained from two-photon microscopic system. It will be specially needed in studying the quantitative three-dimensional information of mouse oocytes.
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Human health risk assessment of pesticide residues in market-sold vegetables and fish in a northern metropolis of China.
Environ Sci Pollut Res Int
PUBLISHED: 09-09-2014
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With growing concerns about food safety and stricter national standards in China, attention has focused on vegetables and fish as they are an important part of the Chinese daily diet, and pesticide residues can accumulate in these foodstuffs. The local consumption habits of vegetables and fish were determined using questionnaires distributed in the major regions of the northern metropolis. Then, the samples of fruit-like vegetables, leafy and root vegetables, and five species of fish (freshwater and marine) were collected from supermarkets and traditional farmers' markets in the city. The concentrations and profiles of pesticide residues (hexachlorocyclohexane (HCH), dichlorodiphenyl trichloroethane (DDT), and endosulfan) in the samples were determined and compared. For the vegetables, the concentration ranges of ?DDT, ?HCH, and ?endosulfan were not detectable (ND) to 10.4 ng/g fresh weight (f.w.), ND to 58.8 ng/g f.w., and ND to 63.9 ng/g f.w., respectively. For the fish samples, the corresponding values were 0.77-25.0 ng/g f.w., 0.02-1.42 ng/g f.w., and 1.22-22.1 ng/g f.w., respectively. Only one celery sample exceeded the maximum residue limits (MRLs) of HCH residues set by Chinese regulations (GB2763-2014). The estimated daily intakes (EDIs) and hazard ratios (HRs) were calculated using data from the recently published Exposure Factors Handbook for the Chinese Population. The EDIs and HRs showed that the levels of organochlorine pesticide (OCP) residues in vegetables and fish in this area are safe.
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Hinokitiol reduces matrix metalloproteinase expression by inhibiting Wnt/?-Catenin signaling in vitro and in vivo.
Int. Immunopharmacol.
PUBLISHED: 09-03-2014
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In this study, we investigated the effects of hinokitiol on matrix metalloproteinase (MMP)-1, -3, -13, collagen type II (Col2a1) and ?-catenin expressions in rat chondrocytes induced by interleukin-1? and in an experimental rat model induced by intra-articular injection of mono-iodoacetate (MIA) into the knee.
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[Association between urinary polycyclic aromatic hydrocarbon metabolites and elevated serumuric acid levels in coke oven workers].
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi
PUBLISHED: 08-30-2014
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To analyze the relationship between metabolites of polycyclic aromatic hydrocarbons (PAHs) and serum uric acid levels in coke oven workers and to provide new clues to the pathogenic mechanism of PAHs.
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The interaction of APEX1 variant with polycyclic aromatic hydrocarbons on increasing chromosome damage and lung cancer risk among male Chinese.
Mol. Carcinog.
PUBLISHED: 08-22-2014
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Polycyclic aromatic hydrocarbons (PAHs) are the most significant contributors to tobacco-induced lung carcinogenesis. Apurinic/apyrimidinic endonuclease 1 (APE1) is a central enzyme in the removal of apurinic/apyrimidinic sites caused by DNA damaging agents. This study aimed to investigate the potential interaction of APEX1 polymorphisms and PAHs on genetic damage and lung cancer risk among male Chinese. We recruited an occupational cohort of 922 male coke oven workers and determined their DNA damage levels by calculating the lymphocytic micronucleus (MN) frequencies. Two well-studied APEX1 polymorphisms (-307A?>?C and Asp148Glu) and their associations with MN frequencies were examined. The impact of MN-related single nucleotide polymorphism (SNP) on lung cancer risk was further investigated in two case-control studies including 1634 male lung cancer patients and 1678 controls. It was shown that, the APEX1 148Glu allele was associated with significantly higher MN frequencies than 148Asp allele, with strongest associations among the highest PAH-exposure workers (P?=?0.008). The APEX1 148Glu allele was also associated with increased lung cancer risk among male smokers, especially among heavy smokers in both case-control studies (odd ratio: 4.40, 95%CI: 3.29-5.72). In addition, APEX1 148Glu variant interacts with smoking in increasing male lung cancer risk, as measured by the attributable proportion due to interaction, which was 0.23 (95%CI: 0.06-0.39). This study showed evidence on interaction between APEX1 148Glu variant and cigarette smoking in increasing lung cancer susceptibility among male Chinese, which may be due to the synergistic effects of APEX1 148Glu and PAHs in increasing chromosome damage levels. The results provide a new insight into gene-interactions in lung carcinogenesis. © 2014 Wiley Periodicals, Inc.
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Selective synthesis of polyfunctionalized pyrido[2,3-b]indoles by multicomponent domino reactions.
J. Org. Chem.
PUBLISHED: 08-13-2014
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A series of novel polyfunctionalized pyrido[2,3-b]indoles were synthesized by three- or four-component domino reactions under microwave irradiation. This protocol has the advantages of readily available starting materials, short reaction times, high yields, easy workup, and high chemo- and regioselectivities.
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Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin a (IgA) nephropathy and healthy individuals.
Mol. Cell Proteomics
PUBLISHED: 07-28-2014
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Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. Although total plasma IgA in IgAN patients was elevated ?1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis.
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Soybean whey protein/chitosan complex behavior and selective recovery of kunitz trypsin inhibitor.
J. Agric. Food Chem.
PUBLISHED: 07-15-2014
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Proteins in soybean whey were separated by Tricine-SDS-PAGE and identified by MALDI-TOF/TOF-MS. In addition to ?-amylase, soybean agglutinin (SBA), and Kunitz trypsin inhibitor (KTI), a 12 kDa band was found to have an amino acid sequence similar to that of Bowman-Birk protease inhibitor (BBI) and showed both trypsin and chymotrypsin inhibitor activities. The complex behavior of soybean whey proteins (SWP) with chitosan (Ch) as a function of pH and protein to polysaccharide ratio (RSWP/Ch) was studied by turbidimetric titration and SDS-PAGE. During pH titration, the ratio of zeta potentials (absolute values) for proteins to chitosan (|ZSWP|/ZCh) at the initial point of phase separation (pH?1) was equal to the reciprocal of their mass ratio (SWP/Ch), revealing that the electric neutrality conditions were fulfilled. The maximum protein recovery (32%) was obtained at RSWP/Ch = 4:1 and pH 6.3, whereas at RSWP/Ch = 20:1 and pH 5.5, chitosan consumption was the lowest (0.196 g Ch/g recovered proteins). In the protein-chitosan complex, KTI and the 12 kDa protein were higher in content than SBA and ?-amylase. However, if soybean whey was precentrifuged to remove aggregated proteins and interacted with chitosan at the conditions of SWP/Ch = 100:1, pH 4.8, and low ionic strength, KTI was found to be selectively complexed. After removal of chitosan at pH 10, a high-purity KTI (90% by SEC-HPLC) could be obtained.
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Galectin-4 expression is associated with reduced lymph node metastasis and modulation of Wnt/?-catenin signalling in pancreatic adenocarcinoma.
Oncotarget
PUBLISHED: 07-01-2014
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Galectin-4 (Gal-4) has been recently identified as a pivotal factor in the migratory capabilities of a set of defined pancreatic ductal adenocarcinoma (PDAC) cell lines using zebrafish as a model system. Here we evaluated the expression of Gal-4 in PDAC tissues selected according to their lymph node metastatic status (N0 vs. N1), and investigated the therapeutic potential of targeting the cross-link with the Wnt signaling pathway in primary PDAC cells. Analysis of Gal-4 expression in PDACs showed high expression of Gal-4 in 80% of patients without lymph node metastasis, whereas 70% of patients with lymph node metastases had low Gal-4 expression. Accordingly, in primary PDAC cells high Gal-4 expression was negatively associated with migratory and invasive ability in vitro and in vivo. Knockdown of Gal-4 in primary PDAC cells with high Gal-4 expression resulted in significant increase of invasion (40%) and migration (50%, P<0.05), whereas enforced expression of Gal-4 in primary cells with low Gal-4 expression reduced the migratory and invasive behavior compared to the control cells. Gal-4 markedly reduces ?-catenin levels in the cell, counteracting the function of Wnt signaling, as was assessed by down-regulation of survivin and cyclin D1. Furthermore, Gal-4 sensitizes PDAC cells to the Wnt inhibitor ICG-001, which interferes with the interaction between CREB binding protein (CBP) and ?-catenin. Collectively, our data suggest that Gal-4 lowers the levels of cytoplasmic ?-catenin, which may lead to lowered availability of nuclear ?-catenin, and consequently diminished levels of nuclear CBP-?-catenin complex and reduced activation of the Wnt target genes. Our findings provide novel insights into the role of Gal-4 in PDAC migration and invasion, and support the analysis of Gal-4 for rational targeting of Wnt/?-catenin signaling in the treatment of PDAC.
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Measuring temporal dynamics of resting-state fMRI data.
Biomed Mater Eng
PUBLISHED: 06-24-2014
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Resting state functional MRI (rs-fMRI), which is used to measure blood oxygen level-dependent (BOLD) from resting brains, is a relatively new and powerful method for evaluating regional interactions that occur when a participant is not performing an explicit task. Because of the sensitiveness to the phase shift and length of time courses of the BOLD recordings, region of interest based conventional correlation and coherence methods are no longer suitable for rs-fMRI analyses. In this paper, we propose a more robust and consistent method, dominant frequency mapping, to analyze rs-fMRI data. We found a dominant frequency of BOLD recordings, 0.0137 Hz, in resting human brains that is consistent across participants and brain regions. This frequency is detected mainly in Gyrus Rectus, Frontal Medial Orbital, Frontal Superior Orbital and Olfactory Sulcus, which control the human social behavior, emotion, and decision making. In the meantime, we found that BOLD frequencies are most inconsistent in the brain regions of PrecentralGyrus, Superior Frontal gyrus, Insula, Caudate nucleus, Putamen, and part of the cerebellum, whose functions are about motor.
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Perspective on polychlorinated dibenzo-p-dioxin and dibenzofuran emissions during chemical production in China: an overlooked source of contemporary relevance.
Environ Sci Pollut Res Int
PUBLISHED: 06-07-2014
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Polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/DFs) are pollutants of significant global concern, and China with its large size and industries is one of the main dioxin-emitting countries in the world. PCDDs/DFs may be formed during the manufacture of chemicals and can either remain in the products as impurities or be emitted into the environment or residues disposed to landfills. The uncertainties in the environmental emissions of PCDDs/DFs from the chemical production industry in China are large because of the complex nature of the industry and variability in the technologies used and limited monitoring conducted. In the current study, we used the PCDD/DF emission factor from the updated United Nations Environment Programme (UNEP) toolkit 2013, information from otherwise published data, and the chemical production data in 2010 to estimate PCDD/DF emissions from the chemical productions in China. Based on these data, it was estimated that there is 1480 g toxic equivalent (TEQ) from the chemical production industry in China, which is much higher than the value that was estimated and used in the national implementation plans (NIPs) for China (102.4 g TEQ in 2004). These results indicate that current PCDD/DF emissions from the chemical production industry in China may be overlooked. Therefore, we suggest that attention should be paid to PCDD/DF emissions from the chemical production industry in future updates of the Chinese NIP and that appropriate measures to decrease PCDD/DF emissions should be taken by better monitoring of products and processes in chemical production industry.
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Density functional study on the structural, electronic, and magnetic properties of 3d transition-metal-doped Au5 clusters.
J Phys Chem A
PUBLISHED: 05-27-2014
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Density functional calculations have been performed for the structural, electronic, and magnetic properties of Au5M (M = Sc-Zn) clusters. Geometry optimizations indicate that the M atoms in low-energy Au5M isomers prefer to occupy the most highly coordinated position. The ground-state clusters except Au5Sc possess a planar structure. The vibrational spectra of the doped clusters are completely different from that of a pure gold cluster. The relative stability and chemical activity are investigated through the averaged binding energy and energy gap for the most stable Au5M clusters. It is found that the impurity atoms (not including the Zn atom) can enhance the thermal stability of the host cluster. The chemical activity of Au5M clusters is higher than that of the Au6 cluster. The calculated energy gaps are in accord with available approximate experimental data. The vertical ionization potential, the electron affinity, and photoelectron spectrum are computed and simulated theoretically for all of the ground-state clusters. The magnetism analyses show that the magnetic moment of these Au5M clusters varies from 0 to 5 ?B by substituting a Au atom in a Au6 cluster with various M atoms and is mainly localized on the M atom for M = Ti-Ni.
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Fucose-based PAMPs prime dendritic cells for follicular T helper cell polarization via DC-SIGN-dependent IL-27 production.
Nat Commun
PUBLISHED: 05-26-2014
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Dendritic cells (DCs) orchestrate antibody-mediated responses to combat extracellular pathogens including parasites by initiating T helper cell differentiation. Here we demonstrate that carbohydrate-specific signalling by DC-SIGN drives follicular T helper cell (TFH) differentiation via IL-27 expression. Fucose, but not mannose, engagement of DC-SIGN results in activation of IKK?, which collaborates with type I IFNR signalling to induce formation and activation of transcription factor ISGF3. Notably, ISGF3 induces expression of IL-27 subunit p28, and subsequent IL-27 secreted by DC-SIGN-primed DCs is pivotal for the induction of Bcl-6(+)CXCR5(+)PD-1(hi)Foxp1(lo) TFH cells, IL-21 secretion by TFH cells and T-cell-dependent IgG production by B cells. Thus, we have identified an essential role for DC-SIGN-induced ISGF3 by fucose-based PAMPs in driving IL-27 and subsequent TFH polarization, which might be harnessed for vaccination design.
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S5a binds to death receptor-6 to induce THP-1 monocytes to differentiate through the activation of the NF-?B pathway.
J. Cell. Sci.
PUBLISHED: 05-14-2014
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Analyses of supernatants from apoptotic cells have helped in the identification of many signals that modulate the states of cell activation and differentiation. However, the current knowledge about the soluble factors that are released during apoptosis is rather limited. Previous studies have shown that S5a and angiocidin (both encoded by PSMD4) induce human acute monocytic leukemia cells (THP-1 cells) to differentiate into macrophages, but the cell-surface receptor of S5a has not been identified. In this study, we show that apoptotic THP-1 cells release endogenous S5a that binds to death receptor-6 (DR6, also known as TNFRSF1), which was identified as an orphan receptor, to induce THP-1 cells to differentiate. Furthermore, we found that the NF-?B pathway is activated, and that the transcription factors WT1 (Wilms' tumor 1) and c-myb mediate S5a-induced THP-1 differentiation. We also show that differentiation is blocked by anti-DR6 antibody, DR6 siRNA, DR6-Fc, NF-?B inhibitor or WT1 siRNA treatment. Our findings indicate that the interaction between cells can determine their differentiation, and we provide evidence for a functional interaction between S5a and DR6, which provides a novel potential mechanism to induce the differentiation of cancer cells, especially during biotherapy for leukemia.
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Overexpression of stathmin 1 is a poor prognostic biomarker in non-small cell lung cancer.
Lab. Invest.
PUBLISHED: 05-12-2014
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Stathmin 1 (STMN1), a major microtubule-depolymerizing protein, is involved in cell cycle progression and cell motility. However, the clinical significance of STMN1 expression in non-small cell lung cancer (NSCLC) has not been determined. The expression pattern of STMN1 mRNA was analyzed by quantitative real-time PCR (qRT-PCR) in 37 cases of NSCLC and in the corresponding non-tumor tissue samples. Furthermore, immunohistochemistry was performed to detect STMN1 protein expression in 113 primary NSCLC tissues. The functional role of STMN1 in lung cancer cell lines was evaluated by small interfering RNA-mediated depletion followed by analyses of cell proliferation and invasion. We found that the STMN1 mRNA and protein levels in NSCLC tissues were significantly higher than those in the corresponding non-tumor tissues (P<0.001). In addition, increased STMN1 expression was correlated with poor tumor differentiation (P<0.001), large tumor size (P=0.022), advanced N stage (P=0.033), and advanced TNM stage (P<0.001). Kaplan-Meier analysis indicates that NSCLC patients with higher STMN1 expression showed significantly worse survival. Moreover, multivariate analysis indicates that higher STMN1 protein expression was an independent prognostic factor of disease-specific survival (HR 2.247, 95%CI 1.320-3.825, P=0.003). Finally, the knockdown of STMN1 in lung cancer cells resulted in a decrease in cellular proliferation and invasion. Our findings suggest that STMN1 may have an important role in NSCLC progression and could serve as a potential prognostic marker for patients with NSCLC.Laboratory Investigation advance online publication, 10 November 2014; doi:10.1038/labinvest.2014.124.
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Application of neural networks with novel independent component analysis methodologies to a Prussian blue modified glassy carbon electrode array.
Talanta
PUBLISHED: 05-06-2014
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Sodium potassium absorption ratio (SPAR) is an important measure of agricultural water quality, wherein four exchangeable cations (K(+), Na(+), Ca(2+) and Mg(2+)) should be simultaneously determined. An ISE-array is suitable for this application because its simplicity, rapid response characteristics and lower cost. However, cross-interferences caused by the poor selectivity of ISEs need to be overcome using multivariate chemometric methods. In this paper, a solid contact ISE array, based on a Prussian blue modified glassy carbon electrode (PB-GCE), was applied with a novel chemometric strategy. One of the most popular independent component analysis (ICA) methods, the fast fixed-point algorithm for ICA (fastICA), was implemented by the genetic algorithm (geneticICA) to avoid the local maxima problem commonly observed with fastICA. This geneticICA can be implemented as a data preprocessing method to improve the prediction accuracy of the Back-propagation neural network (BPNN). The ISE array system was validated using 20 real irrigation water samples from South Australia, and acceptable prediction accuracies were obtained.
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A possible mechanism in DHEA-mediated protection against osteoarthritis.
Steroids
PUBLISHED: 05-04-2014
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Dehydroepiandrosterone (DHEA) and its ester form, DHEA-S, are the most abundant steroids in human plasma. Our previous studies showed that DHEA protects against osteoarthritis (OA). The aim of this paper was to explore the possible mechanisms that underlie DHEA-mediated protection against OA. We tested the expression of ?-catenin, it was increased significantly in OA. Rabbit cartilage was treated with various concentrations of DHEA in both IL-1?-induced rabbit chondrocytes and in rabbit cartilage from the anterior cruciate ligament transaction-induced OA model. We found DHEA decreased the expression of ?-catenin. Then we further activated Wnt/?-catenin signaling by ?-catenin transfection and inactivated it by the inhibitor Dickkopf1 in chondrocytes to reveal its role in the pathogenesis of OA. It turns out the protective effect of DHEA was significantly decreased when Wnt/?-catenin signaling was activated, while inactivating Wnt/?-catenin signaling enhanced the effects of DHEA. Therefore, we hypothesize that DHEA probably exerted its chondroprotective effect by regulating Wnt/?-catenin signaling. Our findings demonstrate the critical role of Wnt/?-catenin signaling in DHEA-mediated protection against OA.
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Differential expression of anti-glycan antibodies in schistosome-infected humans, rhesus monkeys and mice.
Glycobiology
PUBLISHED: 04-11-2014
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Schistosomiasis is a debilitating parasitic disease of humans, endemic in tropical areas, for which no vaccine is available. Evidence points to glycan antigens as being important in immune responses to infection. Here we describe our studies on the comparative humoral immune responses to defined schistosome-type glycan epitopes in Schistosoma mansoni-infected humans, rhesus monkeys and mice. Rhesus anti-glycan responses over the course of infection were screened on a defined glycan microarray comprising semi-synthetic glycopeptides terminating with schistosome-associated or control mammalian-type glycan epitopes, as well as a defined glycan microarray of mammalian-type glycans representing over 400 glycan structures. Infected rhesus monkeys generated a high immunoglobulin G (IgG) antibody response to the core xylose/core ?3 fucose epitope of N-glycans, which peaked at 8-11 weeks post infection, coinciding with maximal ability to kill schistosomula in vitro. By contrast, infected humans generated low antibody levels to this epitope. At 18 months following praziquantel therapy to eliminate the parasite, antibody levels were negligible. Mice chronically infected with S. mansoni generated high levels of anti-fucosylated LacdiNAc (GalNAc?1, 4(Fuc?1, 3)GlcNAc) IgM antibodies, but lacked a robust response to the core xylose/core ?3 fucose N-glycan antigens compared with other species studied, and their sera demonstrated an intermediate level of schistosomula killing in vitro. These differential responses to parasite glycan antigens may be related to the ability of rhesus monkeys to self-cure in contrast to the chronic infection seen in humans and mice. Our results validate defined glycan microarrays as a useful technology to evaluate diagnostic and vaccine antigens for schistosomiasis and perhaps other infections.
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Hereditary breast and ovarian cancer and reproduction: an observational study on the suitability of preimplantation genetic diagnosis for both asymptomatic carriers and breast cancer survivors.
Breast Cancer Res. Treat.
PUBLISHED: 04-01-2014
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Preimplantation genetic diagnosis (PGD) is a reproductive option for BRCA1/2 mutation carriers wishing to avoid transmission of the predisposition for hereditary breast and ovarian cancer (HBOC) to their offspring. Embryos obtained by in vitro fertilisation (IVF/ICSI) are tested for the presence of the mutation. Only BRCA-negative embryos are transferred into the uterus. The suitability and outcome of PGD for HBOC are evaluated in an observational cohort study on treatments carried out in two of Western-Europe's largest PGD centres from 2006 until 2012. Male carriers, asymptomatic female carriers and breast cancer survivors were eligible. If available, PGD on embryos cryopreserved before chemotherapy was possible. Generic PGD-PCR tests were developed based on haplotyping, if necessary combined with mutation detection. 70 Couples underwent PGD for BRCA1/2. 42/71 carriers (59.2 %) were female, six (14.3 %) of whom have had breast cancer prior to PGD. In total, 145 PGD cycles were performed. 720 embryos were tested, identifying 294 (40.8 %) as BRCA-negative. Of fresh IVF/PGD cycles, 23.9 % resulted in a clinical pregnancy. Three cycles involved PGD on embryos cryopreserved before chemotherapy; two of these women delivered a healthy child. Overall, 38 children were liveborn. Two BRCA1 carriers were diagnosed with breast cancer shortly after PGD treatment, despite negative screening prior to PGD. PGD for HBOC proved to be suitable, yielding good pregnancy rates for asymptomatic carriers as well as breast cancer survivors. Because of two cases of breast cancer shortly after treatment, maternal safety of IVF(PGD) in female carriers needs further evaluation.
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Deletion of the CAP10 gene of Cryptococcus neoformans results in a pleiotropic phenotype with changes in expression of virulence factors.
Res. Microbiol.
PUBLISHED: 03-31-2014
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The human pathogen Cryptococcus neoformans causes meningo-encephalitis. The polysaccharide capsule is an important virulence factor for this yeast-like fungus. Previously, we had shown that disruption of the CAP10 gene, encoding a putative xylosyltransferase, results in mutant cells that lack most of the capsular polysaccharides on the cell surface, but do not show a typical acapsular phenotype. In contrast to the acapsular cap59 mutant, cap10 did not induce maturation of dendritic cells when exposed to components of the immune system. In order to gain further insight into the causes of this phenotype displayed by the cap10 mutant, we performed a more in-depth phenotypic analysis of the cell wall and surface structures of this mutant compared to the wild type strain and acapsular mutant cap59. Moreover, we analyzed the cap10 mutant and the wild type strain for differential gene expression of, amongst others, enzymes that are involved in biogenesis of cell wall and capsule components. We conclude that a mutation in the CAP10 gene results in a pleiotropic phenotype with effects on different cellular processes affecting, amongst others, cell size, expression of virulence factors and size of extracellular vesicles.
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Concentration and health risk evaluation of heavy metals in market-sold vegetables and fishes based on questionnaires in Beijing, China.
Environ Sci Pollut Res Int
PUBLISHED: 03-26-2014
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Concentrations of heavy metals (As, Cd, Pb, Cu, Ni, Fe, Mn, and Zn) in market vegetables and fishes in Beijing, China, are investigated, and their health risk to local consumers is evaluated by calculating the target hazard quotient (THQ). The heavy metal concentrations in vegetables and fishes ranged from not detectable (ND) to 0.21 mg/kg fresh weight (f.w.) (As), ND to 0.10 mg/kg f.w. (Cd), and n.d to 0.57 mg/kg f.w. (Pb), with average concentrations of 0.17, 0.04, and 0.24 mg/kg f.w., respectively. The measured concentrations of As, Cd, Pb, Cu, Ni, Fe, Mn, and Zn are generally lower than the safety limits given by the Chinese regulation safety and quality standards of agriculture products (GB2762-2012). As, Cd, and Pb contaminations are found in vegetables and fishes. The exceeding standard rates are 19 % for As, 3 % for Cd, and 25 % for Pb. Pb contaminations are found quite focused on the fish samples from traditional agri-product markets. The paper further analyzed the health risk of heavy metals in vegetables and fishes respectively from supermarkets and traditional agri-product markets; the results showed that the fishes of traditional agri-product markets have higher health risk, while the supermarkets have vegetables of higher heavy metal risk, and the supervision should be strengthened in the fish supply channels in traditional agri-product markets.
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Evidence for increased SOX3 dosage as a risk factor for X-linked hypopituitarism and neural tube defects.
Am. J. Med. Genet. A
PUBLISHED: 03-24-2014
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Genomic duplications of varying lengths at Xq26-q27 involving SOX3 have been described in families with X-linked hypopituitarism. Using array-CGH we detected a 1.1 Mb microduplication at Xq27 in a large family with three males suffering from X-linked hypopituitarism. The duplication was mapped from 138.7 to 139.8 Mb, harboring only two annotated genes, SOX3 and ATP11C, and was shown to be a direct tandem copy number gain. Unexpectedly, the microduplication did not fully segregate with the disease in this family suggesting that SOX3 duplications have variable penetrance for X-linked hypopituitarism. In the same family, a female fetus presenting with a neural tube defect was also shown to carry the SOX3 copy number gain. Since we also demonstrated increased SOX3 mRNA levels in amnion cells derived from an unrelated t(X;22)(q27;q11) female fetus with spina bifida, we propose that increased levels of SOX3 could be a risk factor for neural tube defects.
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Plasma microRNA expression and micronuclei frequency in workers exposed to polycyclic aromatic hydrocarbons.
Environ. Health Perspect.
PUBLISHED: 03-13-2014
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Ubiquitous polycyclic aromatic hydrocarbons (PAHs) have been shown to alter gene expression patterns and elevate micronuclei (MN) frequency, but the underlying mechanisms are largely unknown. MicroRNAs (miRNAs) are key gene regulators that may be influenced by PAH exposures and mediate their effects on MN frequency.
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Circulating MicroRNAs and the occurrence of acute myocardial infarction in Chinese populations.
Circ Cardiovasc Genet
PUBLISHED: 03-13-2014
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Circulating microRNAs ( miRNAs) are emerging as novel disease biomarkers. We aimed to explore the association between circulating miRNAs and the occurrence of acute myocardial infarction (AMI) in Chinese populations.
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Polycyclic aromatic hydrocarbons-associated microRNAs and their interactions with the environment: influences on oxidative DNA damage and lipid peroxidation in coke oven workers.
Environ. Sci. Technol.
PUBLISHED: 03-12-2014
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We previously identified five polycyclic aromatic hydrocarbons (PAHs)-associated microRNAs (miRNAs) and found they were associated with chromosome damage. As oxidative damage is the common contributory cause of various PAHs-related diseases, we further investigated the influences of these miRNAs and their interactions with environmental factors on oxidative DNA damage and lipid peroxidation. We measured PAHs internal exposure biomarkers [urinary monohydroxy-PAHs (OH-PAHs) and plasma benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydotetrol-albumin (BPDE-Alb) adducts], the expression levels of PAHs-associated plasma miRNAs (miR-24-3p, miR-27a-3p, miR-142-5p, miR-28-5p, and miR-150-5p), and urinary biomarkers of oxidative DNA damage [8-hydroxydeoxyguanosine (8-OH-dG)] and lipid peroxidation [8-iso-prostaglandin-F2? (8-iso-PGF2?)] in 365 healthy male coke oven workers. These miRNAs were associated with a dose-response increase in 8-OH-dG (? > 0), and with a dose-response decrease in 8-iso-PGF2? (? < 0), especially in workers with lower PAHs exposure levels, in nonsmokers, and in nondrinkers. These miRNAs interacted antagonistically with ?OH-PAHs and BPDE-Alb adducts (?interaction < 0) and synergistically with drinking status (?interaction > 0) to influence 8-OH-dG, while they interacted synergistically with BPDE-Alb adducts (?interaction > 0) and antagonistically with smoking status (?interaction < 0) to influence 8-iso-PGF2?. Our results suggested that miRNAs and their interactions with environmental factors might be novel mechanisms mediating the effects of PAHs exposure on oxidative DNA damage and lipid peroxidation.
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Long non-coding RNAs in colorectal cancer: implications for pathogenesis and clinical application.
Mod. Pathol.
PUBLISHED: 03-07-2014
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Long non-coding RNAs (lncRNAs) are a class of newly identified non-coding RNA molecules that are emerging as key regulators of tumor initiation and development. Colorectal cancer (CRC) remains a major health problem worldwide, and there remains a need to further refine the current screening approaches as well as provide tailored diagnostic and therapeutic approaches. Multiple dysregulated lncRNAs participate in tumorigenesis through a variety of molecular mechanisms, and various regulatory factors frequently contribute to the aberrant expression of lncRNAs in CRC, thereby allowing malignant transformation. Additionally, the association of dysregulated lncRNAs with specific developmental stages and clinical outcomes indicates their potential as strong diagnostic and prognostic predictors as well as therapeutic targets. Here we provide a brief overview of the known functions of CRC-associated lncRNAs, describe some potential molecular mechanisms that underlie changes in lncRNA expression in CRC, and attempt to uncover their clinical and therapeutic potential.
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Decision-making on preimplantation genetic diagnosis and prenatal diagnosis: a challenge for couples with hereditary breast and ovarian cancer.
Hum. Reprod.
PUBLISHED: 03-06-2014
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How do couples with a BRCA1/2 mutation decide on preimplantation genetic diagnosis (PGD) and prenatal diagnosis (PND) for hereditary breast and ovarian cancer syndrome (HBOC)?
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Concentrations, atmospheric partitioning, and air-water/soil surface exchange of polychlorinated dibenzo-p-dioxin and dibenzofuran along the upper reaches of the Haihe River basin, North China.
Environ Sci Pollut Res Int
PUBLISHED: 03-03-2014
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Polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/PCDF) were overall measured and compared in ambient air, water, soils, and sediments along the upper reaches of the Haihe River of North China, so as to evaluate their concentrations, profiles, and to understand the processes of gas-particle partitioning and air-water/soil exchange. The following results were obtained: (1) The average concentrations (toxic equivalents, TEQs) of 2,3,7,8-PCDD/PCDF in air, water, sediment, and soil samples were 4,855 fg/m(3), 9.5 pg/L, 99.2 pg/g dry weight (dw), and 56.4 pg/g (203 fg TEQ/m(3), 0.46 pg TEQ/L, 2.2 pg TEQ/g dw, and 1.3 pg TEQ/g, respectively), respectively. (2) Although OCDF, 1,2,3,4,6,7,8-HpCDF, OCDD, and 1,2,3,4,6,7,8-HpCDD were the dominant congeners among four environmental sinks, obvious discrepancies of these congener and homologue patterns of PCDD/PCDF were observed still. (3) Significant linear correlations for PCDD/PCDF were observed between the gas-particle partition coefficient (K p) and the subcooled liquid vapor pressure (P L (0)) and octanol-air partition coefficient (K oa). (4) Fugacity fraction values of air-water exchange indicated that most of PCDD/PCDF homologues were dominated by net volatilization from water into air. The low-chlorinated PCDD/PCDF (tetra- to hexa-) presented a strong net volatilization from the soil into air, while high-chlorinated PCDD/PCDF (hepta- to octa-) were mainly close to equilibrium for air-soil exchange.
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Excreted/secreted Trichuris suis products reduce barrier function and suppress inflammatory cytokine production of intestinal epithelial cells.
Mol. Immunol.
PUBLISHED: 02-26-2014
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The administration of helminths is considered a promising strategy for the treatment of autoimmune diseases due to their immunomodulatory properties. Currently, the application of the helminth Trichuris suis as a treatment for Crohn's disease is being studied in large multi-center clinical trials. The intestinal epithelium forms an efficient barrier between the intestinal lumen containing the microbial flora and helminths, and dendritic cells (DCs) present in the lamina propria that determine the TH response. Here, we investigated how excreted/secreted (E/S) products of T. suis affect the barrier function of intestinal epithelial cells (IECs) in order to reach the DCs and modulate the immune response. We show that T. suis E/S products reduce the barrier function and the expression of the tight junction proteins EMP-1 and claudin-4 in IEC CMT93/69 monolayers in a glycan-dependent manner. This resulted in an increased passage of soluble compounds to the basolateral side that affected DC function. In addition, T. suis E/S suppressed LPS-induced pro-inflammatory cytokine production by CMT93/69 cells, whereas the production of the TH2 response-inducing cytokine thymic stromal lymphopoietin (TSLP) was induced. Our studies indicate that T. suis E/S glycans affect the function of the intestinal epithelium in order to modulate DC function. Identification of the T. suis E/S glycans that modulate IEC and DC function may lead to a strategy to reduce symptoms of autoimmune and allergic immune diseases by orally administrated helminth-derived factors without the need of infection with live helminths.
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Nicotine exposure increases the complexity of dopamine neurons in the parainterfascicular nucleus (PIF) sub-region of VTA.
J Neuroeng Rehabil
PUBLISHED: 02-25-2014
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Recent publications highlight differences within the sub-regions of the ventral tegmental area (VTA) including the parabrachial pigmented nucleus (PBP), parainterfascicular nucleus (PIF) and paranigral nucleus (PN) in the projections to the prefrontal cortex (PFC) and the glutamatergic pathway.
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An FBN1 deep intronic mutation in a familial case of Marfan syndrome: an explanation for genetically unsolved cases?
Hum. Mutat.
PUBLISHED: 02-21-2014
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Marfan syndrome (MFS) is caused by mutations in the FBN1 (fibrillin-1) gene, but approximately 10% of MFS cases remain genetically unsolved. Here, we report a new FBN1 mutation in an MFS family that had remained negative after extensive molecular genomic DNA FBN1 testing, including denaturing high-performance liquid chromatography, Sanger sequencing, and multiplex ligation-dependent probe amplification. Linkage analysis in the family and cDNA sequencing of the proband revealed a deep intronic point mutation in intron 56 generating a new splice donor site. This mutation results in the integration of a 90-bp pseudo-exon between exons 56 and 57 containing a stop codon, causing nonsense-mediated mRNA decay. Although more than 90% of FBN1 mutations can be identified with regular molecular testing at the genomic level, deep intronic mutations will be missed and require cDNA sequencing or whole-genome sequencing.
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The helminth Trichuris suis suppresses TLR4-induced inflammatory responses in human macrophages.
Genes Immun.
PUBLISHED: 02-20-2014
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Recent clinical trials in patients with inflammatory diseases like multiple sclerosis (MS) or inflammatory bowel disease (IBD) have shown the beneficial effects of probiotic helminth administration, although the underlying mechanism of action remains largely unknown. Potential cellular targets may include innate immune cells that propagate inflammation in these diseases, like pro-inflammatory macrophages. We here investigated the effects of the helminth Trichuris suis soluble products (SPs) on the phenotype and function of human inflammatory (granulocyte-macrophage colony-stimulating factor (GM-CSF)-differentiated) macrophages. Interestingly, we here show that T. suis SPs potently skew inflammatory macrophages into a more anti-inflammatory state in a Toll-like receptor 4 (TLR4)-dependent manner, and less effects are seen when stimulating macrophages with TLR2 or -3 ligands. Gene microarray analysis of GM-CSF-differentiated macrophages further revealed that many TLR4-induced inflammatory mediators, including interleukin (IL)-12B, CCL1 and CXCL9, are downregulated by T. suis SPs. In particular, we observed a strong reduction in the expression and function of P2RX7, a purinergic receptor involved in macrophage inflammation, leading to reduced IL-1? secretion. In conclusion, we show that T. suis SPs suppress a broad range of inflammatory pathways in GM-CSF-differentiated macrophages in a TLR4-dependent manner, thereby providing enhanced mechanistic insight into the therapeutic potential of this helminth for patients with inflammatory diseases.
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Catalytic synthesis of sulfated polysaccharides. II: comparative studies of solution conformation and antioxidant activities.
Carbohydr Polym
PUBLISHED: 02-20-2014
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Sulfated derivatives of Artemisia sphaerocephala polysaccharide (ASP) with high degree of substitution (DS) were synthesized using 4-dimethylaminopyridine (DMAP)/dimethylcyclohexylcarbodiimide (DCC) as catalyst. Size exclusion chromatography combined with multi-angle laser light scattering (SEC-LLS) results showed a decrease in fractal dimension (df) values of sulfated ASP (SASP). Compared to ASP and SASP with low DS (0.51-1.01), SASPcata2 exhibited an internal structure between rigid rod and random coil with a DS of 1.24. DS had greater influence on its conformation in aqueous solution. Circular dichroism (CD), methylene blue (MB) and Congo red (CR) spectrophotometric method and atomic force microscopy (AFM) results confirmed that the degradation of ASP and SO3H groups improved significantly the stiffness of the chains due to the electrostatic effect. Furthermore, antioxidant experiments revealed that high DS could enhance the scavenging activities of radicals and reducing power of SASP in vitro. The extended chain conformation was beneficial to enhance the biological activity of sulfated polysaccharides.
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A genome-wide association study identifies common variants influencing serum uric acid concentrations in a Chinese population.
BMC Med Genomics
PUBLISHED: 02-05-2014
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Uric acid (UA) is a complex phenotype influenced by both genetic and environmental factors as well as their interactions. Current genome-wide association studies (GWASs) have identified a variety of genetic determinants of UA in Europeans; however, such studies in Asians, especially in Chinese populations remain limited.
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Endogenous endothelial progenitor cells participate in neovascularization via CXCR4/SDF-1 axis and improve outcome after stroke.
CNS Neurosci Ther
PUBLISHED: 01-17-2014
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To study whether endogenous endothelial progenitor cells (EPCs) are involved in neovascularization after stroke.
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Exploration of disulfide bridge and N-glycosylation contributing to high thermostability of a hybrid xylanase.
Protein Pept. Lett.
PUBLISHED: 01-12-2014
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A comparison between three-dimensional structures of a wild-type xylanase AoXyn11A and a hybrid xylanase AEx11A revealed that a disulfide bridge (Cys(5)-Cys(32)) and an N-glycosylation site (Asn(42)) were imported into AEx11A by N-terminal substitution of AoXyn11A with EvXyn11(TS). Two mutant genes AEx11A(C5T) and AEx11A(N42Q) were constructed by mutating Cys(5)- and Asn(42)-encoding codons of AEx11A into Thr(5)- and Gln(42)-encoding ones, and heterologously expressed in Pichia pastoris GS115, respectively. The temperature optimum of the recombinant AEx11A(C5T) (reAEx11A(C5T)) was decreased to 60°C from 80°C of reAEx11A, while its thermal inactivation half-lives at 70 and 80°C shortened to 3 and 1 min from 197 and 25 min of reAEx11A, respectively. However, there was no obvious alteration between reAEx11A and reAEx11A(C5T) in pH characteristics and kinetic parameters. Furthermore, both reAEx11A(N42Q) and reAEx11A displayed no significant difference in all enzymatic properties tested, except for the apparent molecular weight. We concluded based on this study that the disulfide bridge of AEx11A was vital to its high thermostability, but the N-glycosylation had no effect on.
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Down-regulation of ncRAN, a long non-coding RNA, contributes to colorectal cancer cell migration and invasion and predicts poor overall survival for colorectal cancer patients.
Mol. Carcinog.
PUBLISHED: 01-07-2014
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Accumulating evidence has indicated that long non-coding RNAs (lncRNAs) play critical roles in regulating cellular processes, such as cell growth and apoptosis, as well as cancer progression and metastasis. ncRAN (non-coding RNA expressed in aggressive neuroblastoma) was previously shown to be dramatically up-regulated and associated with poor prognosis in human neuroblastoma. This lncRNA also plays an important role in bladder cancer growth and invasion. Colorectal cancer (CRC) progression typically follows a complex cascade from primary malignancy to distant metastasis, but whether the aberrant expression of ncRAN in CRC is associated with malignancy, metastasis or prognosis remains unknown. In this study, we demonstrated that ncRAN expression is significantly down-regulated in tumor tissue and CRC cell lines compared with adjacent normal tissue and a normal intestinal mucous cell line. Reduced expression of ncRAN was detected in poorly differentiated or undifferentiated tumors and in tumors with liver metastases. Kaplan-Meier analysis indicated that patients with lower ncRAN expression have a worse overall survival. Moreover, multivariate analysis revealed that decreased expression of ncRAN is an independent predictor of overall survival. Our experimental data indicated that ncRAN mediates the in vitro migration and invasion of CRC cells. Together, these results suggest that ncRAN might represent a novel prognostic indicator, a biomarker for the early detection of metastasis and a target for gene therapy in CRC. © 2014 Wiley Periodicals, Inc.
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BcPMI2, isolated from non-heading Chinese cabbage encoding phosphomannose isomerase, improves stress tolerance in transgenic tobacco.
Mol. Biol. Rep.
PUBLISHED: 01-04-2014
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Phosphomannose isomerase (PMI) is an enzyme that catalyses the first step of the L-galactose pathway for ascorbic acid (AsA) biosynthesis in plants. To clarify the physiological roles of PMI in AsA biosynthesis, the cDNA sequence of PMI was cloned from non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) and overexpressed in tobacco transformed with Agrobacterium tumefaciens. The AsA and soluble sugar contents were lower in 35S::BcPMI2 tobacco than in wild-type tobacco. However, the AsA level in BcPMI2-overexpressing plants under stress was significantly increased. The T1 seed germination rate of transgenic plants was higher than that of wild-type plants under NaCl or H2O2 treatment. Meanwhile, transgenic plants showed higher tolerance than wild-type plants. This finding implied that BcPMI2 overexpression improved AsA biosynthetic capability and accumulation, and evidently enhanced tolerance to oxidative and salt stress, although the AsA level was lower in transgenic tobacco than in wild-type tobacco under normal condition.
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Discriminating males and unpredictable females: males differentiate self-similar facial cues more than females in the judgment of opposite-sex attractiveness.
PLoS ONE
PUBLISHED: 01-01-2014
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Attractiveness judgment in the context of mate preferences is thought to reflect an assessment of mate quality in relation to an absolute scale of genetic fitness and a relative scale of self-similarity. In this study, subjects judged the attractiveness and trustworthiness of faces in composite images that were manipulated to produce self-similar (self-resemblance) and dissimilar (other-resemblance) images. Males differentiated between self- and other-resemblance as well as among different degrees of self-resemblance in their attractiveness ratings; females did not. Specifically, in Experiment 1, using a morphing technique, we created previously unseen face images possessing different degrees (0%, 30%, 40%, or 50%) of incorporation of the subject's images (different degrees of self-resemblance) and found that males preferred images that were closer to average (0%) rather than more self-similar, whereas females showed no preference for any degree of self-similarity. In Experiment 2, we added a pro-social question about trustworthiness. We replicated the Experiment 1 attractiveness rating results and further found that males differentiated between self- and other-resemblance for the same degree of composites; women did not. Both males and females showed a similar preference for self-resemblances when judging trustworthiness. In conclusion, only males factored self-resemblance into their attractiveness ratings of opposite-sex individuals in a manner consistent with cues of reproductive fitness, although both sexes favored self-resemblance when judging trustworthiness.
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Galactofuranose-Coated Gold Nanoparticles Elicit a Pro-inflammatory Response in Human Monocyte-Derived Dendritic Cells and Are Recognized by DC-SIGN.
ACS Chem. Biol.
PUBLISHED: 12-09-2013
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Galactofuranose (Galf) is the five-membered ring form of galactose exclusively found in nonmammalian species, among which several are pathogens. To determine the putative role of this carbohydrate in host-pathogen interactions, we synthesized multivalent gold nanoparticles carrying Galf (Galf-GNPs) and show that they are recognized by the EB-A2 antibody, which is widely used to detect Galf-containing galactomannan in the serum of Aspergillosis patients. We demonstrated that human monocyte-derived dendritic cells bound Galf-GNPs via interaction with the lectin DC-SIGN. Moreover, interaction of dendritic cells with Galf-GNPs resulted in increased expression of several maturation markers on these cells and induced secretion of the pro-inflammatory cytokines IL-6 and TNF-?. These data indicate that Galf is able to modulate the innate immune response via dendritic cells. In conclusion, Galf-GNPs are a versatile tool that can be applied in multiple functional studies to gain a better understanding of the role of Galf in host-pathogen interaction.
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PLS3 mutations in X-linked osteoporosis with fractures.
N. Engl. J. Med.
PUBLISHED: 10-02-2013
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Plastin 3 (PLS3), a protein involved in the formation of filamentous actin (F-actin) bundles, appears to be important in human bone health, on the basis of pathogenic variants in PLS3 in five families with X-linked osteoporosis and osteoporotic fractures that we report here. The bone-regulatory properties of PLS3 were supported by in vivo analyses in zebrafish. Furthermore, in an additional five families (described in less detail) referred for diagnosis or ruling out of osteogenesis imperfecta type I, a rare variant (rs140121121) in PLS3 was found. This variant was also associated with a risk of fracture among elderly heterozygous women that was two times as high as that among noncarriers, which indicates that genetic variation in PLS3 is a novel etiologic factor involved in common, multi-factorial osteoporosis.
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[Construction of a recombinant adenovirus co-expressing bone morphogenic proteins 9 and 6 and its effect on osteogenesis in C3H10 cells].
Nan Fang Yi Ke Da Xue Xue Bao
PUBLISHED: 09-27-2013
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To construct a recombinant adenovirus co-expressing bone morphogenic protein (BMP) 9 and BMP6 and observe its effect on the osteogenesis in C3H10 cells.
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Genome-wide association study on serum alkaline phosphatase levels in a Chinese population.
BMC Genomics
PUBLISHED: 09-24-2013
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Serum alkaline phosphatase (ALP) is a complex phenotype influenced by both genetic and environmental factors. Recent Genome-Wide Association Studies (GWAS) have identified several loci affecting ALP levels; however, such studies in Chinese populations are limited. We performed a GWAS analyzing the association between 658,288 autosomal SNPs and serum ALP in 1,461 subjects, and replicated the top SNPs in an additional 8,830 healthy Chinese Han individuals. The interactions between significant locus and environmental factors on serum ALP levels were further investigated.
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Reprogramming macrophages to an anti-inflammatory phenotype by helminth antigens reduces murine atherosclerosis.
FASEB J.
PUBLISHED: 09-16-2013
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Atherosclerosis is a lipid-driven inflammatory disease of the vessel wall, characterized by the chronic activation of macrophages. We investigated whether the helminth-derived antigens [soluble egg antigens (SEAs)] could modulate macrophage inflammatory responses and protect against atherosclerosis in mice. In bone marrow-derived macrophages, SEAs induce anti-inflammatory macrophages, typified by high levels of IL-10 and reduced secretion of proinflammatory mediators. In hyperlipidemic LDLR(-/-) mice, SEA treatment reduced plaque size by 44%, and plaques were less advanced compared with PBS-injected littermate controls. The atheroprotective effect of SEAs was found to be mainly independent of cholesterol lowering and T-lymphocyte responses but instead could be attributed to diminished myeloid cell activation. SEAs reduced circulating neutrophils and inflammatory Ly6C(high) monocytes, and macrophages showed high IL-10 production. In line with the observed systemic effects, atherosclerotic lesions of SEA-treated mice showed reduced intraplaque inflammation as inflammatory markers [TNF-?, monocyte chemotactic protein 1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and CD68], neutrophil content, and newly recruited macrophages were decreased. We show that SEA treatment protects against atherosclerosis development by dampening inflammatory responses. In the future, helminth-derived components may provide novel opportunities to treat chronic inflammatory diseases, as they diminish systemic inflammation and reduce the activation of immune cells.-Wolfs, I. M. J., Stöger, J. L., Goossens, P., Pöttgens, C., Gijbels, M. J. J., Wijnands, E., van der Vorst, E. P. C., van Gorp, P., Beckers, L., Engel, D., Biessen, E. A. L., Kraal, G., van Die, I., Donners, M. M. P. C., de Winther, M. P. J. Reprogramming macrophages to an anti-inflammatory phenotype by helminth antigens reduces murine atherosclerosis.
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Clinical and health costs impact of progress in diagnosis and treatment in venous thromboembolic disease: evolution in 15 years.
Ann Vasc Surg
PUBLISHED: 08-15-2013
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The therapeutic and diagnostic approach in deep vein thrombosis (DVT) has changed enormously in the last two decades with the introduction of ultrasound, low-molecular-weight heparin (LMWH), and premature motion. The aim of this study is to evaluate these changes and analyze their clinical and economic aspects.
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Proteomic analysis of non-heading Chinese cabbage infected with Hyaloperonospora parasitica.
J Proteomics
PUBLISHED: 08-11-2013
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Downy mildew is a serious fungal disease in non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) that is caused by Hyaloperonospora parasitica, which infects members of the Brassicaceae family. For breeding improvement, researchers must understand the defence mechanisms employed by non-heading Chinese cabbage to combat H. parasitica infection. Using 2-DE protein analysis, we compared the proteomes from leaves of non-heading Chinese cabbage seedlings that were infected with H. parasitica or that were only treated with water at different time points post-infection. By MS analysis, 91 protein spots with significant differences in abundance (>2-fold, p<0.05) were identified in mock- and H. parasitica-inoculated leaves. Next, a resistance strategy for incompatible interactions was proposed. This network consisted of several functional components, including enhanced ethylene biosynthesis and energy supply, balanced ROS production and scavenging, accelerated protein metabolism and photorespiratory, reduced photosynthesis, and induced photosystem repair. These findings increase our knowledge of incompatible interactions between plants and pathogens and also provide new insight regarding the function of plant molecular processes, which should assist in the discovery of new strategies for pathogen control.
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[Mechanism of reductive dechlorination of trichlorophenol with different electron donors].
Huan Jing Ke Xue
PUBLISHED: 08-07-2013
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A test was conducted to examine the degradation effect and reductive dechlorination pathway of 2, 4, 6-trichlorophenol (2,4,6-TCP) in the presence of different electron donors, such as glucose, sodium lactate, sodium pyruvate and sodium acetate. The results showed that, compared with the effect of glucose, sodium lactate, sodium pyruvate and sodium acetate enhanced the dechlorination of 2, 4, 6-TCP effectively, among which sodium lactate could serve as a kind of hydrogen release compound, and the electrons required for reductive dechlorination were released in a sustained way. Substrate metabolism dehydrogenase activity was improved by the external electron donor; after reaction for 240 h, the activity of dehydrogenase was increased in the four electron donor systems, by 21.49%, 25.78%, 136.85% and 139.3%, respectively. The main reductive dechlorination products of 2,4,6-TCP included 2,4-dichlorophenol (2,4-DCP), 4-chlorophenol (4-CP) and phenol; when sodium acetate was used as the electron donor, 4-CP was the main degradation product, and the transformation ratio from 2,4,6-TCP to 4-CP was more than 22%.
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DCIR interacts with ligands from both endogenous and pathogenic origin.
Immunol. Lett.
PUBLISHED: 07-29-2013
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C-type lectins on dendritic cells function as antigen uptake and signaling receptors, thereby influencing cellular immune responses. Dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) is one of the best-studied C-type lectin receptors expressed on DCs and its glycan specificity and functional requirements for ligand binding have been intensively investigated. The carbohydrate specificity of dendritic cell immunoreceptor (DCIR), another DC-expressed lectin, was still debated, but we have recently confirmed DCIR as mannose/fucose-binding lectin. Since DC-SIGN and DCIR may potentially share ligands, we set out to elucidate the interaction of DCIR with established DC-SIGN-binding ligands, by comparing the carbohydrate specificity of DCIR and DC-SIGN in more detail. Our results clearly demonstrate that DC-SIGN has a broader glycan specificity compared to DCIR, which interacts only with mannotriose, sulfo-Lewis(a), Lewis(b) and Lewis(a). While most of the tested DC-SIGN ligands bound DCIR as well, Candida albicans and some glycoproteins on some cancer cell lines were identified as DC-SIGN-specific ligands. Interestingly, DCIR strongly bound human immunodeficiency virus type 1 (HIV-1) gp140 glycoproteins, while its interaction with the well-studied DC-SIGN-binding HIV-1 ligand gp120 was much weaker. Furthermore, DCIR-specific ligands were detected on keratinocytes. Furthermore, the interaction of DCIR with its ligands was strongly influenced by the glycosylation of DCIR. In conclusion, we show that sulfo-Lewis(a) is a high affinity ligand for DCIR and that DCIR interacts with ligands from both pathogenic and endogenous origin of which most are shared by DC-SIGN.
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Functional analysis of the distal region of the third intracellular loop of PROKR2.
Biochem. Biophys. Res. Commun.
PUBLISHED: 07-29-2013
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Mutations in the G-protein-coupled receptor PROKR2 have been identified in patients with idiopathic hypogonadotropic hypogonadism (IHH) and Kallmann syndrome (KS) manifesting with delayed puberty and infertility. Recently, the homozygous mutation V274D was identified in a man displaying KS with an apparent reversal of hypogonadism. The affected amino acid, valine 274, is located at the junction region of the third intracellular loop (IL3) and the sixth transmembrane domain (TM6). In this study, we first studied the effect of V274D and related mutations (V274A, V274T, and V274R) on the signaling activity and cell surface expression of PROKR2. Our data indicate that a charged amino acid substitution at residue 274 of PROKR2 results in low cell surface expression and loss-of-function. Furthermore, we studied the effects of two clusters of basic amino acids located at the proximal region of Val274 on the cell surface expression and function of PROKR2. The deletion of RRK (270-272) resulted in undetectable cell surface expression, whereas RKR (264-266)-deleted PROKR2 was expressed normally on the cell surface but showed loss-of-function due to a deficiency in G-protein coupling. Our data indicate that the distal region of the IL3 of PROKR2 may differentially influence receptor trafficking and G-protein coupling.
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Lactococcus lactis anchoring avian infectious bronchitis virus multi-epitope peptide EpiC induced specific immune responses in chickens.
Biosci. Biotechnol. Biochem.
PUBLISHED: 07-07-2013
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Mucosal immunity is critical in preventing infectious bronchitis virus (IBV) infection. To deliver viral antigens to the mucosal immune system of chickens safely and effectively, we constructed a Lactococcus lactis strain carrying IBV multi-epitope gene EpiC fused with the gene of the cell-wall anchoring domain of Staphylococcus aureus protein A. SDS-PAGE and Western blot results indicated that the fused peptide was located partially on the cell surface. Oral and nasal inoculation with the recombinant L. lactis of chickens elicited significantly high humoral and mucosal immune responses, especially in the nasally immunized group. Eighty percent chickens of the nasally immunized group with recombinant L. lactis did not show any clinical signs after a lethal dose challenge with IBV SAIBk strain, while all the non-recombinant L. lactis immunized chickens exhibited obvious and typical symptoms. These results indicate that needle-free recombinant lactococci anchoring the IBV antigen makes a promising vaccine candidate against the spread of IB.
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Changes in drug sensitivity and anti-malarial drug resistance mutations over time among Plasmodium falciparum parasites in Senegal.
Malar. J.
PUBLISHED: 07-06-2013
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Malaria treatment efforts are hindered by the rapid emergence and spread of drug resistant parasites. Simple assays to monitor parasite drug response in direct patient samples (ex vivo) can detect drug resistance before it becomes clinically apparent, and can inform changes in treatment policy to prevent the spread of resistance.
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Interaction of smoothened with integrin-linked kinase in primary cilia mediates Hedgehog signalling.
EMBO Rep.
PUBLISHED: 07-01-2013
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Here we report that ILK localizes in the mouse primary cilium, a sensory organelle required for signalling by the Hedgehog (Hh) pathway. Genetic or pharmacological inhibition of ILK blocks ciliary accumulation of the Hh pathway effector smoothened (Smo) and suppresses the induction of Gli transcription factor mRNAs by SHh. Conditional deletion of ILK or Smo also inhibits SHh-driven activation of Gli2 in the embryonic mouse cerebellum. ILK regulation of Hh signalling probably requires the physical interaction of ILK and Smo in the cilium, and we also show selective cilia-associated interaction of ILK with ?-arrestin, a known mediator of Smo-dependent signalling.
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Carotenogenic gene expression and carotenoid accumulation in three varieties of Cucurbita pepo during fruit development.
J. Agric. Food Chem.
PUBLISHED: 06-21-2013
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The control of gene expression is a crucial regulatory mechanism in carotenoid accumulation of fruits and flowers. We investigated the role of transcriptional regulation of nine genes involved in the carotenoid biosynthesis pathway in three varieties of Cucurbita pepo with evident differences in fruit color. The transcriptional levels of the key genes involved in the carotenoid biosynthesis were higher in flower-, leaf-, and fruit skin tissues than flesh tissues. This correlated with higher concentration of carotenoid content in these tissues. The differential expression among the colored and white cultivars detected for some genes, such as LCYe, in combination with other regulatory mechanisms, could explain the large differences found in terms of carotenoid content among the three varieties. These results are a first step to elucidate carotenogenesis in C. pepo and demonstrate that, in general, regulation of the pathway genes is a critical factor that determines the accumulation of these compounds.
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Dose-response relationships of polycyclic aromatic hydrocarbons exposure and oxidative damage to DNA and lipid in coke oven workers.
Environ. Sci. Technol.
PUBLISHED: 06-21-2013
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Polycyclic aromatic hydrocarbons (PAHs) are known to induce reactive oxygen species and oxidative stress, but the dose-response relationships between exposure to PAHs and oxidative stress levels have not been established. In this study, we recruited 1333 male coke oven workers, monitored the levels of environmental PAHs, and measured internal PAH exposure biomarkers including 12 urinary PAH metabolites and plasma benzo[a]pyrene-r-7,t-8,t-9,c-10-tetrahydotetrol-albumin (BPDE-Alb) adducts, as well as the two oxidative biomarkers urinary 8-hydroxydeoxyguanosine (8-OHdG) and 8-iso-prostaglandin-F2? (8-iso-PGF2?). We found that the total concentration of urinary PAH metabolites and plasma BPDE-Alb adducts were both significantly associated with increased 8-OHdG and 8-iso-PGF2? in both smokers and nonsmokers (all p < 0.05). This exposure-response effect was also observed for most PAH metabolites (all p(trend) < 0.01), except for 4-hydroxyphenanthrene and 8-OHdG (p(trend) = 0.108). Furthermore, it was shown that only urinary 1-hydroxypyrene has a significant positive association with both 8-OHdG and 8-iso-PGF2? after a Bonferroni correction (p < 0.005). Our results indicated that urinary ?OH-PAHs and plasma BPDE-Alb adducts can result in significant dose-related increases in oxidative damage to DNA and lipids. Furthermore, when a multianalyte method is unavailable, our findings demonstrate that urinary 1-hydroxypyrene is a useful biomarker for evaluating total PAHs exposure and assessing oxidative damage in coke oven workers.
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Glycosyltransferases in chemo-enzymatic synthesis of oligosaccharides.
Methods Mol. Biol.
PUBLISHED: 06-15-2013
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Many oligosaccharides are not commercially available, which limits studies focused on elucidation of glycan functions; therefore chemo-enzymatic methods to synthesize them can be very useful. Here, we describe the procedure to synthesize the Gal?1-3GalNAc?1-4GlcNAc?-R (Gal-LDN) moiety, containing the Gal?1-3GalNAc epitope found on the parasitic helminth Haemonchus contortus. An acceptor substrate providing a terminal N-acetylglucosamine was prepared by coupling the fluorescent hydrophobic aglycon, 2,6-diaminopyridine (DAP), to N,N-diacetylchitobiose. By the subsequent action of recombinant Caenorhabditis elegans ?1,4-N-acetylgalactosaminyltransferase the substrate was efficiently converted to GalNAc?1-4GlcNAc?-R (LDN-R). Since no recombinant ?1,3-galactosyltransferase has been described that acts on terminal ?GalNAc, we used bovine ?1,3-galactosyltransferase to obtain a partial conversion of LDN-R to the Gal-LDN antigen. This method can be applied to synthesize any oligosaccharide, provided that specific glycosyltransferases are available, or related enzymes that can be pushed to elongate the selected acceptor.
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Glycans from avian influenza virus are recognized by chicken dendritic cells and are targets for the humoral immune response in chicken.
Mol. Immunol.
PUBLISHED: 06-13-2013
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To increase our understanding of the interaction between avian influenza virus and its chicken host, we identified receptors for putative avian influenza virus (AIV) glycan determinants on chicken dendritic cells. Chicken dendritic cells (DCs) were found to recognize glycan determinants containing terminal ?GalNAc, Gal?1-3Gal, GlcNAc?1-4GlcNAc?1-4GlcNAc? (chitotriose) and Gal?1-2Gal. Infection of chicken dendritic cells with either low pathogenic (LP) or highly pathogenic (HP) AIV results in elevated mRNA expression of homologs of the mouse C-type lectins DEC205 and macrophage mannose receptor (MMR), whereas expression levels of the human dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) homolog remained unchanged. Following uptake and subsequent presentation of avian influenza virus by DCs, adaptive immunity, including humoral immune responses are induced. We have investigated the antibody responses against virus glycan epitopes after avian influenza virus infection. Using glycan micro-array analysis we showed that chicken contained antibodies that predominantly recognize terminal Gal?1-3Gal-R, chitotriose and Fuc?1-2Gal?1-4GlcNAc-R (H-type 2). After influenza-infection, glycan array analysis showed that both levels and repertoire of glycan-recognizing antibodies decreased. However, analysis of the sera by ELISA indicated that the levels of different isotypes of anti-glycan Abs against specific glycan antigens was increased after influenza-infection, suggesting that the presentation of the glycan antigens and iso-type of the Abs are critical parameters to take into account when measuring anti-glycan Abs. This novel approach in avian influenza research may contribute to the development of a broad spectrum vaccine and improves our mechanistic understanding of innate and adaptive responses to glycans.
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Do we really need closed-suction drainage in total hip arthroplasty? A meta-analysis.
Int Orthop
PUBLISHED: 05-22-2013
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The clinical use of closed-suction drainage, which aims to reduce postoperative wound haematomas and infection, is common. This study was performed to determine whether closed-suction drainage is safe and effective in promoting wound healing and reducing blood loss and other complications compared with no-drainage in total hip arthroplasty.
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Brain white matter oedema due to ClC-2 chloride channel deficiency: an observational analytical study.
Lancet Neurol
PUBLISHED: 05-22-2013
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Mutant mouse models suggest that the chloride channel ClC-2 has functions in ion and water homoeostasis, but this has not been confirmed in human beings. We aimed to define novel disorders characterised by distinct patterns of MRI abnormalities in patients with leukoencephalopathies of unknown origin, and to identify the genes mutated in these disorders. We were specifically interested in leukoencephalopathies characterised by white matter oedema, suggesting a defect in ion and water homoeostasis.
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Tetrandrine induces apoptosis and triggers a caspase cascade in U2-OS and MG-63 cells through the intrinsic and extrinsic pathways.
Mol Med Rep
PUBLISHED: 05-16-2013
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Although neoadjuvant chemotherapy has improved the survival rate of osteosarcoma patients, drug resistance remains a predominant obstacle to improving efficacy and necessitates the development of novel chemotherapeutical agents. The aim of this study was to investigate whether tetrandrine (TET) induces apoptosis in the U-2OS and MG-63 osteosarcoma cell lines and to further determine the underlying mechanism. This study investigated the effects of TET on osteosarcoma in vitro. To examine the antitumor effects of TET on osteosarcoma, the two osteosarcoma cell lines were treated with TET and subjected to apoptosis assays. The results revealed that TET induced the apoptosis of osteosarcoma cells in a time- and dose-dependent manner. Furthermore, the apoptosis of osteosarcoma cells was accompanied by increased cytochrome c (Cyto-C), apoptotic protease-activating factor (Apaf)-1, Bid and Bax activation and reduced Bcl-2 and Bcl-xl activation, demonstrating that the apoptosis may have occurred through the mitochondrial pathway. In conclusion, the results suggest that TET is a promising agent for osteosarcoma therapy.
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Imputation-based association analyses identify new lung cancer susceptibility variants in CDK6 and SH3RF1 and their interactions with smoking in Chinese populations.
Carcinogenesis
PUBLISHED: 05-03-2013
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Cell cycle regulation, apoptosis, oxidative stress and inflammation response play critical roles in the development of smoking-induced lung cancer. However, it is still not well known whether their genetic variants are associated with lung cancer susceptibility. In this study, we performed imputation-based association analyses to investigate the influence of common genetic variants in these pathways and their interactions with smoking on lung cancer susceptibility. We first selected 24 042 unvalidated genetic variants in 798 genes from the imputed dataset of the previous lung cancer genome-wide association study in 2331 cases and 3077 controls, and then conducted additional two-stage validations in 4133 cases and 4522 controls. We found a genome-wide significant (P < 5.0 × 10(-8)) association for rs2282987 in CDK6 at 7q21.2 [odds ratio (OR) = 1.18, combined P add = 2.27 × 10(-9)] and a consistent association for rs2706748 in SH3RF1 at 4q32.3 (OR = 1.17, combined P add = 5.10 × 10(-6)). Interaction analyses showed that rs2282987 and rs2706748 interacted with both smoking status (P interaction were 1.04 × 10(-2) and 3.03 × 10(-2), respectively) and smoking history (P interaction were 1.21 × 10(-2) and 5.21 × 10(-2), respectively) to contribute to lung cancer susceptibility in subjects aged 51-60 years. These results further underscore the contribution of genetic variants involved in pathways of cell cycle regulation and apoptosis to lung cancer susceptibility, and highlight gene-environment interactions in lung cancer etiology, especially in subjects aged 51-60 years.
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Occupational Exposure to Formaldehyde and Genetic Damage in the Peripheral Blood Lymphocytes of Plywood Workers.
J Occup Health
PUBLISHED: 05-02-2013
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Objectives: We sought to clarify the association of occupational formaldehyde exposure with DNA strand breaks, chromosome damage and DNA-protein crosslinks (DPCs) in the peripheral blood (PB) lymphocytes of plywood workers. Methods: We determined Olive tail moment (OTM) values, micronucleus (MN) frequencies and DPC rates of the PB lymphocytes in 178 workers divided into control and lower and higher exposure groups according to their current formaldehyde exposure levels and examined the association of each end point with formaldehyde exposure levels and with the number of work years. We also examined each end point in an additional 62 workers before and after an 8-hour formaldehyde exposure for validating the association. Results: OTM values increased significantly in the two exposure groups compared with those in the control group (P < 0.05 for both) and were associated with increasing formaldehyde exposure levels (Ptrend = 0.002), while MN frequencies increased with increasing numbers of work years (Ptrend < 0.001). The dynamic study showed that OTM values and DPC rates increased after an 8-hour formaldehyde exposure compared with those before the exposure (P < 0.001, P = 0.019, respectively), that, in a dose-dependent manner, OTM values were associated with formaldehyde exposure levels during work hours (P = 0.005) and that MN frequencies before and after the 8-hour work exposure were associated with numbers of work years (P = 0.029, P < 0.001, respectively). Conclusions: We found a dose-response relationship between the current formaldehyde exposure levels and DNA strand breaks and between duration of exposure and chromosome damage in the PB lymphocytes of plywood workers.
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Passive Movement Improves the Learning and Memory Function of Rats with Cerebral Infarction by Inhibiting Neuron Cell Apoptosis.
Mol. Neurobiol.
PUBLISHED: 04-23-2013
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Passive movement has been found to improve evidently ischemic stroke patients impaired capacity of learning and memory, but the optimal time window of initiating the therapy and the underlying mechanism are not fully understood. In this study, the effect of passive movement at different time windows on learning and memory of rats with cerebral infarction was detected. The results showed that the expression of caspase-3 and escape latency in the passive movement group were all considerably lower than those in the model group (P?
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Postural orthostatic tachycardia syndrome with increased erythrocytic hydrogen sulfide and response to midodrine hydrochloride.
J. Pediatr.
PUBLISHED: 04-03-2013
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To evaluate the use of erythrocytic hydrogen sulfide (H2S) in predicting the therapeutic efficacy of midodrine hydrochloride for children with postural orthostatic tachycardia syndrome (POTS).
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ATF3 suppresses metastasis of bladder cancer by regulating gelsolin-mediated remodeling of the actin cytoskeleton.
Cancer Res.
PUBLISHED: 03-27-2013
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Bladder cancer is associated with high recurrence and mortality rates due to metastasis. The elucidation of metastasis suppressors may offer therapeutic opportunities if their mechanisms of action can be elucidated and tractably exploited. In this study, we investigated the clinical and functional significance of the transcription factor activating transcription factor 3 (ATF3) in bladder cancer metastasis. Gene expression analysis revealed that decreased ATF3 was associated with bladder cancer progression and reduced survival of patients with bladder cancer. Correspondingly, ATF3 overexpression in highly metastatic bladder cancer cells decreased migration in vitro and experimental metastasis in vivo. Conversely, ATF3 silencing increased the migration of bladder cancer cells with limited metastatic capability in the absence of any effect on proliferation. In keeping with their increased motility, metastatic bladder cancer cells had increased numbers of actin filaments. Moreover, ATF3 expression correlated with expression of the actin filament severing protein gelsolin (GSN). Mechanistic studies revealed that ATF3 upregulated GSN, whereas ATF3 silencing reduced GSN levels, concomitant with alterations in the actin cytoskeleton. We identified six ATF3 regulatory elements in the first intron of the GSN gene confirmed by chromatin immunoprecipitation analysis. Critically, GSN expression reversed the metastatic capacity of bladder cancer cells with diminished levels of ATF3. Taken together, our results indicate that ATF3 suppresses metastasis of bladder cancer cells, at least in part through the upregulation of GSN-mediated actin remodeling. These findings suggest ATF3 coupled with GSN as prognostic markers for bladder cancer metastasis.
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Prenatal testing for Huntingtons disease in the Netherlands from 1998 to 2008.
Clin. Genet.
PUBLISHED: 03-27-2013
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This study aims to give an overview of the number of prenatal tests for Huntingtons disease (HD), test results, and pregnancy outcomes in the Netherlands between 1998 and 2008 and to compare them with available data from the period 1987 to 1997. A total of 126 couples underwent prenatal diagnosis (PND) on 216 foetuses: 185 (86%) direct tests and 31 (14%) exclusion tests. In 9% of direct tests the risk for the foetus was 25%. Four at-risk parents (4%) carried intermediate alleles. Ninety-one foetuses had CAG expansions ?36% or 50% risk haplotypes: 75 (82%) were terminated for HD, 12 (13%) were carried to term; four pregnancies were miscarried, terminated for other reasons or lost to follow-up. Unaffected pregnancies (122 foetuses) resulted in the birth of 112 children. The estimated uptake of PND was 22% of CAG expansion carriers (?36 repeats) at reproductive age. PND was used by two new subgroups: carriers of intermediate alleles and 50% at-risk persons opting for a direct prenatal test of the foetus. A significant number of HD expansion or 50% risk pregnancies were continued. Speculations were made on causative factors contributing to these continuations. Further research on these couples motives is needed.
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Low expression of LOC285194 is associated with poor prognosis in colorectal cancer.
J Transl Med
PUBLISHED: 03-22-2013
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BACKGROUND: The long non-coding RNAs (lncRNAs) study has gradually become one of the hot topics in the field of RNA biology. One lncRNA which has attracted attention is LOC285194, a lncRNA demonstrated the potential tumor-suppressor role in osteosarcoma. The aim of this study was to examine the expression of LOC285194 in colorectal cancer (CRC) patients and to investigate the relationship between this lncRNA levels and existing clinicopathologic parameters and patient survival. METHODS: The expression of LOC285194 was detected by quantitative real-time polymerase chain reaction in pairs of tumorous and adjacent normal tissues of 81 colorectal cancer patients with a follow-up of 5 years, as well as in three colorectal cancer cell lines and normal intestinal mucous cell line. Then, we analyzed the potential relationship between this lncRNA levels in tumor tissues and existing clinicopathological features of CRC, and clinical outcome. RESULTS: The relative expression levels of LOC285194 was significantly lower in tumor tissues (p < 0.001) and colorectal cancer cell lines compared with adjacent normal tissues and normal intestinal mucous cell line. In addition, low expression of LOC285194 was correlated with larger tumor size (p = 0.015), higher tumor stage (p = 0.034), and more distant metastasis (p = 0.046). Kaplan-Meier analysis indicated that patients with low LOC285194 expression had a poor disease free survival (p = 0.010). Moreover, multivariate analysis showed that decreased expression of LOC285194 was an independent predictor of disease-specific survival. CONCLUSION: Our data indicate that LOC285194 might be a novel prognostic indicator in colorectal cancer and may be a potential target for diagnosis and gene therapy.
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The uptake and outcome of prenatal and pre-implantation genetic diagnosis for Huntingtons disease in the Netherlands (1998-2008).
Clin. Genet.
PUBLISHED: 03-21-2013
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We aimed to study reproductive behaviour of couples opting for prenatal diagnosis (PND) and pre-implantation genetic diagnosis (PGD) for Huntingtons disease (HD). In the Netherlands, exclusion PND is available for persons at 50% risk, whereas exclusion PGD is not allowed. All 162 couples who underwent PND or PGD for HD between 1998 and 2008 and referrals for exclusion PGD to Belgium were included. Couples reproductive information was collected until December 2010; 132 couples (81.5%) underwent PND in 262 pregnancies, 54 (33.3%) started PGD, and 25 used both. Sixteen percent of PND couples used exclusion PND and 6% used exclusion PGD. The outcomes were 76.5% of PND couples delivered ?1 unaffected child(ren) after PND, and 44.4% of PGD couples delivered ?1 PGD child(ren) (mean 2.5?cycles/couple). Couples opting for PGD secondarily (after a previous pregnancy) had more frequently terminated a pregnancy for HD (87.0%) compared with couples secondarily opting for PND (55.2%; p?=?0.015). At-risk or HD expansion carrier males were underrepresented in the group of couples primarily opting for PGD (25%) and overrepresented in the secondary PGD group (64%). We conclude that couples reconsider their choices in every subsequent pregnancy based on their previous experience, personal beliefs and the gender of the at-risk partner.
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