JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
Ultrasensitive electrochemical biosensing for DNA using quantum dots combined with restriction endonuclease.
Analyst
PUBLISHED: 11-20-2014
Show Abstract
Hide Abstract
A universal and sensitive electrochemical biosensing platform for the detection and identification of DNA using CdSe quantum dots (CdSe QDs) as signal markers was designed. The detection mechanism was based on the specific recognition of MspI endonuclease combined with the signal amplification of gold nanoparticles (AuNPs). MspI endonuclease could recognize its specific sequence in the double-strand DNA (dsDNA) and cleave the dsDNA fragments linked with CdSe QDs from the electrode. The remaining attached CdSe QDs can be easily read out by square-wave voltammetry using an electrodeposited bismuth (Bi) film-modified glass carbon electrode. The concentrations of target DNA could be simultaneously detected by the signal of metal markers. Using mycobacterium tuberculosis (Mtb) DNA as a model, under the optimal conditions, the proposed biosensor could detect Mtb DNA down to 8.7 × 10(-15) M with a linear range of 5 orders of magnitude (from 1.0 × 10(-14) to 1.0 × 10(-9) M) and discriminate mismatched DNA with high selectivity. This strategy presented a universal and convenient biosensing platform for DNA assay, and its satisfactory performances make it a potential candidate for the early diagnosis of gene-related diseases.
Related JoVE Video
Post-synthesis DNA modifications using a trans-cyclooctene click handle.
Org. Biomol. Chem.
PUBLISHED: 11-20-2014
Show Abstract
Hide Abstract
Post-synthesis DNA modification is a very useful method for DNA functionalization. This is achieved by using a modified NTP, which has a handle for further modifications, replacing the corresponding natural NTP in polymerase-catalyzed DNA synthesis. Subsequently, the handle can be used for further functionalization after PCR, preferably through a very fast reaction. Herein we describe polymerase-mediated incorporation of trans-cyclooctene modified thymidine triphosphate (TCO-TTP). Subsequently, the trans-cyclooctene group was reacted with a tetrazine tethered to other functional groups through a very fast click reaction. The utility of this DNA functionalization method was demonstrated with the incorporation of a boronic acid group and a fluorophore. The same approach was also successfully used in modifying a known aptamer for fluorescent labelling applications.
Related JoVE Video
Dianthraceno[a,e]pentalenes: synthesis, crystallographic structures and applications in organic field-effect transistors.
Chem. Commun. (Camb.)
PUBLISHED: 11-20-2014
Show Abstract
Hide Abstract
Two soluble and stable dianthraceno[a,e]pentalenes with two () and six () phenyl substituents were synthesized. Both compounds possess a small energy band gap and show amphoteric redox behaviour due to intramolecular donor-accepter interactions. X-ray crystallographic analysis revealed that has a closely packed structure with multi-dimensional [C-H?] interactions although there are no ?-? interactions between the dianthraceno[a,e]pentalene cores. As a result, solution-processed field effect transistors based on exhibited an average hole mobility of 0.65 cm(2) V(-1) s(-1). Under similar conditions, showed an average field effect hole mobility of 0.001 cm(2) V(-1) s(-1).
Related JoVE Video
High-throughput mapping of brain-wide activity in awake and drug-responsive vertebrates.
Lab Chip
PUBLISHED: 11-20-2014
Show Abstract
Hide Abstract
The reconstruction of neural activity across complete neural circuits, or brain activity mapping, has great potential in both fundamental and translational neuroscience research. Larval zebrafish, a vertebrate model, has recently been demonstrated to be amenable to whole brain activity mapping in behaving animals. Here we demonstrate a microfluidic array system ("Fish-Trap") that enables high-throughput mapping of brain-wide activity in awake larval zebrafish. Unlike the commonly practiced larva-processing methods using a rigid gel or a capillary tube, which are laborious and time-consuming, the hydrodynamic design of our microfluidic chip allows automatic, gel-free, and anesthetic-free processing of tens of larvae for microscopic imaging with single-cell resolution. Notably, this system provides the capability to directly couple pharmaceutical stimuli with real-time recording of neural activity in a large number of animals, and the local and global effects of pharmacoactive drugs on the nervous system can be directly visualized and evaluated by analyzing drug-induced functional perturbation within or across different brain regions. Using this technology, we tested a set of neurotoxin peptides and obtained new insights into how to exploit neurotoxin derivatives as therapeutic agents. The novel and versatile "Fish-Trap" technology can be readily unitized to study other stimulus (optical, acoustic, or physical) associated functional brain circuits using similar experimental strategies.
Related JoVE Video
Mutations in the ABCA3 gene are associated with cataract-microcornea syndrome.
Invest. Ophthalmol. Vis. Sci.
PUBLISHED: 11-20-2014
Show Abstract
Hide Abstract
Cataract-microcornea syndrome (CCMC) is an autosomal dominant inherited disease characterized by the association of congenital cataract and microcornea without any other systemic anomaly or dysmorphism. Although mutations of several genes have been shown to cause dominant CCMC, in many patients the causative gene has not yet been identified. Our aim was to identify the disease-associated gene in Chinese patients with CCMC.
Related JoVE Video
A click-and-release approach to CO prodrugs.
Chem. Commun. (Camb.)
PUBLISHED: 11-07-2014
Show Abstract
Hide Abstract
Carbon monoxide belongs to the family of signaling molecules and has been shown to possess therapeutic effects. Similar to NO, safe delivery of CO is a key issue in developing CO-based therapeutics. Herein we report a "click and release" CO-prodrug approach, which allows the release of CO under physiological conditions without the need for light irradiation. The system releases CO in a triggered and controllable manner and possesses the potential of tunable release rates.
Related JoVE Video
Expression of RBMX in the Light-Induced Damage of Rat Retina In Vivo.
Cell. Mol. Neurobiol.
PUBLISHED: 10-17-2014
Show Abstract
Hide Abstract
RNA-binding motif protein, X-linked (RBMX) is a 43 kDa nuclear protein in the RBM family and functions on alternative splicing of RNA. The gene encoding RBMX is located on chromosome Xq26. To investigate whether RBMX is involved in retinal neuron apoptosis, we performed a light-induced retinal damage model in adult rats. Western blotting analysis showed RBMX gradually increased, reached a peak at 12 h and then declined during the following days. The association of RBMX in retinal ganglion cells (RGCs) with light exposure was found by immunofluorescence staining. The injury-induced expression of RBMX was detected in active caspase-3 and TUNEL positive cells. We also examined the expression profiles of active caspase-3, bcl-2 and Bax, whose changes were correlated with the expression profiles of RBMX. To summarize, we uncovered the dynamic changes of RBMX in the light-induced retinal damage model for the first time. RBMX might play a significant role in the degenerative process of RGCs after light-induced damage in the retina.
Related JoVE Video
[Simultaneous determination of six perfluorinated organic compounds in feed by using polyamide solid-phase extraction with ultra performance liquid chromatography-tandem mass spectrometry].
Se Pu
PUBLISHED: 09-27-2014
Show Abstract
Hide Abstract
A method for the determination of six perfluorinated organic compounds (PFCs) in feed has been developed. It is based on polyamide solid-phase extraction (SPE) together with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The sample was extracted by acidified acetonitrile. The extraction solution was enriched by a polyamide SPE cartridge under acidic condition, and cleaned-up using methanol, eluted by 5% (v/v) ammonia/methanol solvent and determined by UPLC-MS/MS. The UPLC separation was carried out on an Acquity BEH C18 column (100 mm x 2.1 mm, 1.7 microm). The mobile phases were 5 mmol/L ammonium acetate and acetonitrile with a gradient elution. Under the optimal conditions, the PFCs were analyzed under multiple reaction monitoring (MRM) mode with negative electrospray ionization. The isotope internal standard method was used to determine the six PFCs, and improve the quantitative accuracy. All of the target compounds exhibited good linearity (r > 0.995) over a concentration range of 0.5-25 microg/L. The detection limits of the six PFCs were all smaller than 0.1 microg/kg. The mean recoveries of the six PFCs were in the range of 94.2% to 108.9% with the relative standard deviations (RSDs) of 1.8% - 8.6% (n = 6). The method for the determination of PFCs in feed is low-cost, favorable effect and suitable for the detection of complex matrix samples.
Related JoVE Video
Roles of rhizosphere and root-derived organic acids in Cd accumulation by two hot pepper cultivars.
Environ Sci Pollut Res Int
PUBLISHED: 09-18-2014
Show Abstract
Hide Abstract
Cultivars of hot pepper (Capsicum annuum L.) have different abilities to accumulate Cd in their fruits. Previously, we suggested that low-Cd cultivars take up more Cd, but can better prevent the Cd translocation from roots to aerial parts. However, the mechanisms involved in those processes are still unclear. In this study, we explored the roles of rhizosphere soil Cd fractions and root secretions of low molecular weight organic acids in the uptake, translocation, and accumulation of Cd in a low-Cd and high-Cd cultivar. The results showed that there was no significant difference in exchangeable Cd between rhizosphere soils of the two cultivars, which might be related to their similar root's Cd uptake ability. The total content of low molecular weight organic acids released from roots of the low-Cd cultivar was almost equal to that released from roots of the high-Cd cultivar at the same Cd level; however, the composition of low molecular weight organic acids were determined by cultivars and Cd exposure levels. In the higher Cd (10 ?M) treatment, the roots of the low-Cd cultivar excreted significantly less tartaric acid and more oxalic and acetic acids than those of the high-Cd cultivar. Additionally, there was no difference in the concentration of citric or succinic acid between the two cultivars. These results indicate that some kinds of low molecular weight organic acids efflux from hot pepper roots played an important role in the difference of Cd accumulation between low- and high-Cd cultivars.
Related JoVE Video
B?cell translocation gene 1 serves as a novel prognostic indicator of hepatocellular carcinoma.
Int. J. Oncol.
PUBLISHED: 09-17-2014
Show Abstract
Hide Abstract
Although the B?cell translocation gene 1 (BTG1) plays an important role in apoptosis and negatively regulates cell proliferation, BTG1 expression in hepatocellular carcinoma (HCC) has not been evaluated. In this study expression analysis of BTG1 was conducted to clarify the role of BTG1 in the initiation of HCC carcinogenesis and progression. BTG1 mRNA expression levels were determined for HCC cell lines and 151 surgical specimen pairs using quantitative real?time reverse transcription polymerase chain reaction (RT?qPCR) assay. The mutational and methylation status of HCC cell lines were analyzed via high resolution melting (HRM) analysis and direct sequencing analysis to elucidate the regulatory mechanisms of BTG1 expression. The expression and distribution of the BTG1 protein in liver tissues were evaluated using immunohistochemistry (IHC). Decreased expression of BTG1 mRNA was confirmed in the majority of HCC cell lines (89%) and clinical HCC tissues (85%) compared with non?cancerous liver tissues. Mutations or promoter hypermethylation were not identified in HCC cell lines. BTG1 mRNA expression levels were not influenced by background liver status. The pattern of BTG1 protein expression was consistent with that of BTG1 mRNA. Downregulation of BTG1 mRNA in HCC was significantly associated with shorter disease?specific and recurrence?free survival rates. Multivariate analysis of disease?specific survival rates identified BTG1 mRNA downregulation as an independent prognostic factor for HCC (hazard ratio 2.12, 95% confidence interval 1.12?4.04, P=0.022). Our results indicate that altered BTG1 expression might affect hepatocarcinogenesis and may represent a novel biomarker for HCC carcinogenesis and progression.
Related JoVE Video
A carbon nanotube/quantum dot based photoelectrochemical biosensing platform for the direct detection of microRNAs.
Chem. Commun. (Camb.)
PUBLISHED: 09-17-2014
Show Abstract
Hide Abstract
A versatile photoelectrochemical biosensing platform was developed based on DNA-CdS quantum dots (QDs) sensitized single-walled carbon nanotubes (SWCNTs)-COOH. Combining with cyclic enzymatic amplification, a convenient, sensitive and specific biosensor for the direct detection of microRNAs (miRNAs) was designed, which provided a novel approach for analysis of miRNAs.
Related JoVE Video
Photoelectrochemical biosensor using enzyme-catalyzed in situ propagation of CdS quantum dots on graphene oxide.
ACS Appl Mater Interfaces
PUBLISHED: 09-08-2014
Show Abstract
Hide Abstract
An innovative photoelectrochemical (PEC) biosensor platform was designed based on the in situ generation of CdS quantum dots (QDs) on graphene oxide (GO) using an enzymatic reaction. Horseradish peroxidase catalyzed the reduction of sodium thiosulfate with hydrogen peroxide to generate H2S, which reacted with Cd(2+) to form CdS QDs. CdS QDs could be photoexcited to generate an elevated photocurrent as a readout signal. This strategy offered a "green" alternative to inconvenient presynthesis procedures for the fabrication of semiconducting nanoparticles. The nanomaterials and assembly procedures were characterized by microscopy and spectroscopy techniques. Combined with immune recognition and on the basis of the PEC activity of CdS QDs on GO, the strategy was successfully applied to a PEC assay to detect carcinoembryonic antigen and displayed a wide linear range from 2.5 ng mL(-1) to 50 ?g mL(-1) and a detection limit of 0.72 ng mL(-1) at a signal-to-noise ratio of 3. The PEC biosensor showed satisfactory performance for clinical sample detection and was convenient for determining high concentrations of solute without dilution. This effort offers a new opportunity for the development of numerous rapid and convenient analytical techniques using the PEC method that may be applied in the design and preparation of various solar-energy-driven applications.
Related JoVE Video
Clinical utility of PDSS2 expression to stratify patients at risk for recurrence of hepatocellular carcinoma.
Int. J. Oncol.
PUBLISHED: 09-03-2014
Show Abstract
Hide Abstract
Identification of novel genetic and epigenetic alterations is required for optimal stratification of patients with hepatocellular carcinoma (HCC) at risk for recurrence and adverse prognosis. Coenzyme Q10 (CoQ10), which mediates apoptosis, is synthesized by prenyl diphosphate synthase subunit 2 (PDSS2). In the present study we evaluated the clinical significance and regulatory mechanisms of PDSS2 expression in HCC. PDSS2 expression levels and those of genes encoding potentially interacting proteins as well as the methylation status of the PDSS2 promoter region were analyzed in HCC cell lines. PDSS2 mRNA levels in 151 pairs of resected specimens were determined to evaluate the association of PDSS2 expression and clinicopathological factors. The expression and distribution of PDSS2 were determined using immunohistochemistry. PDSS2 mRNA expression was decreased in six of nine HCC cell lines and significantly correlated with those of hepatocyte nuclear factor 4?. PDSS2 transcription in HCC cells with decreased PDSS2 expression accompanying hypermethylation was reactivated after treating these cells with a methylation inhibitor. Mean expression levels of PDSS2 mRNA relative to that of uninvolved liver diminished gradually in the order of chronic hepatitis to cirrhosis, and each was significantly higher than those of HCCs. PDSS2 and PDSS2 mRNA levels were consistent. Decreased PDSS2 mRNA levels were detected in HCC tissues of 56 patients, correlated with shorter disease-specific survival, and was identified as an independent prognostic factor. PDSS2 is a putative tumor suppressor, and promoter hypermethylation is a key regulatory mechanism in HCC. Decreased levels of PDSS2 mRNA expression may represent a novel biomarker of HCC.
Related JoVE Video
Dihydropyrimidinase-like 3 is a putative hepatocellular carcinoma tumor suppressor.
J. Gastroenterol.
PUBLISHED: 08-31-2014
Show Abstract
Hide Abstract
Patients with hepatocellular carcinoma (HCC) may relapse after curative resection. Sensitive biomarkers for HCC are required to enhance disease management. Dihydropyrimidinase-like 3 (DPYSL3) suppresses cell proliferation and tumorigenicity of certain malignancies; however, its role in HCC is unknown.
Related JoVE Video
Construction and characterization of novel hirulog variants with antithrombin and antiplatelet activities.
Protein Pept. Lett.
PUBLISHED: 08-30-2014
Show Abstract
Hide Abstract
The RGD sequence was used to design potent hirudin isoform 3 mimetic peptides with both antithrombin activity and antiplatelet aggregation activity. The RGD and proline were inserted between the catalytic active binding domain (D-Phe-Pro-Arg-Pro) on the N-terminus and the anion-binding exosite binding domain (QGDFEPIPEDAYDE) on the Cterminus. Thrombin titration assay and ATP-induced platelet aggregation test revealed that the peptide with the linker RGDWP or RGDGP possessed potent antithrombin and antiplatelet activities, while other peptides without the Pro residue in the linker only showed antithrombin activity. Similar results were obtained in the RGD-containing hirulog-1 variants. Our study indicates that the inserted Pro residue facilitates the exposure of RGD and the binding of the peptide to glycoprotein IIb/IIIa (GPIIb/IIIa). The strategy of combining the RGD sequence and the Pro residue may be used for future designs of bifunctional antithrombotic agents.
Related JoVE Video
Fibroblast Growth Factor 21 Is Regulated by the IRE1?-XBP1 Branch of the Unfolded Protein Response and Counteracts Endoplasmic Reticulum Stress-induced Hepatic Steatosis.
J. Biol. Chem.
PUBLISHED: 08-28-2014
Show Abstract
Hide Abstract
Endoplasmic reticulum (ER) stress activates the adaptive unfolded protein response (UPR) and represents a critical mechanism that underlies metabolic dysfunctions. Fibroblast growth factor 21 (FGF21), a hormone that is predominantly secreted by the liver, exerts a broad range of effects upon the metabolism of carbohydrates and lipids. Although increased circulating levels of FGF21 have been documented in animal models and human subjects with obesity and nonalcoholic fatty liver disease, the functional interconnections between metabolic ER stress and FGF21 are incompletely understood. Here, we report that increased ER stress along with the simultaneous elevation of FGF21 expression were associated with the occurrence of nonalcoholic fatty liver disease both in diet-induced obese mice and human patients. Intraperitoneal administration of the ER stressor tunicamycin in mice resulted in hepatic steatosis, accompanied by activation of the three canonical UPR branches and increased the expression of FGF21. Furthermore, the IRE1?-XBP1 pathway of the UPR could directly activate the transcriptional expression of Fgf21. Administration of recombinant FGF21 in mice alleviated tunicamycin-induced liver steatosis, in parallel with reduced eIF2?-ATF4-CHOP signaling. Taken together, these results suggest that FGF21 is an integral physiological component of the cellular UPR program, which exerts beneficial feedback effects upon lipid metabolism through counteracting ER stress.
Related JoVE Video
Macrophage-secreted IL-8 induces epithelial-mesenchymal transition in hepatocellular carcinoma cells by activating the JAK2/STAT3/Snail pathway.
Int. J. Oncol.
PUBLISHED: 08-21-2014
Show Abstract
Hide Abstract
Macrophages are a major component of the leukocyte infiltrate of tumors and play a pivotal role in the progression of hepatocellular carcinoma (HCC). However, the molecular mechanisms by which macrophages promote HCC invasion are poorly understood. The present study was undertaken to investigate the relationship between macrophages and epithelial-mesenchymal transition (EMT) of HCC. Double-staining immunohistochemistry was used to observe the association between macrophages and EMT markers in clinical HCC samples and it showed that EMT primarily occurred at the edge of the tumor nest, in which infiltrating macrophages were always observed. This indicated that CD68 which is a marker of macrophages, was correlated with EMT marker levels. In addition, after being cultured with macrophages for 24 h, the ability of HCC cells to migrate and invade increased, Snail and N-Cadherin expression was upregulated, and E-Cadherin was downregulated. An antibody array assay was applied to analyze the supernatant of these cultures and it demonstrated IL-8 increased significantly in the macrophage co-culture system. Finally, the role of macrophage-derived IL-8 in the invasion of HCC cells was assayed, and downstream signaling pathways were also investigated. We found that IL-8: i) may induce EMT and promote HCC cell migration and invasion and ii) is associated with the JAK2/STAT3/Snail signaling pathway. Taking together, these findings revealed that macrophages that have infiltrated tumors may induce epithelial-mesenchymal transition of HCC cells via the IL-8 activated JAK2/STAT3/Snail pathway. Thus, this may offer a potential target for developing new HCC therapies.
Related JoVE Video
Root morphological responses of three hot pepper cultivars to Cd exposure and their correlations with Cd accumulation.
Environ Sci Pollut Res Int
PUBLISHED: 08-15-2014
Show Abstract
Hide Abstract
Cultivars of hot pepper (Capsicum annuum L.) differ widely in their fruit cadmium (Cd) concentrations. Previously, we suggested that low-Cd cultivars are better able to prevent the translocation of Cd from roots to aboveground parts, but the corresponding mechanisms are still unknown. In this study, we aimed to improve understanding of the root morphological characteristics of the mechanisms involved in two low-Cd and a high-Cd cultivar. Seedlings were grown in nutrient solutions containing 0 (control), 2, and 10 ?M Cd for 20 days, and Cd contents for the three cultivars were compared with changes in root morphology. The total root length (RL), root surface area (SA), number of root tips (RT), and specific root length (SRL) of all cultivars were decreased significantly by the 10 ?M Cd treatment with the exception of the SA in JFZ, which showed no obvious change. For each cultivar, the 10 ?M Cd treatment decreased significantly RL and SA specifically in roots with diameters (RD) of RD???0.2 mm or 0.2 mm?
Related JoVE Video
A Candidate Plasma Protein Classifier to Identify Alzheimer's Disease.
J. Alzheimers Dis.
PUBLISHED: 08-11-2014
Show Abstract
Hide Abstract
Biomarkers currently used in the aid for the diagnosis of Alzheimer's disease (AD) are cerebrospinal fluid (CSF) protein markers and brain neuroimaging markers. These biomarkers, however, either involve semi-invasive procedures or are costly to measure. Thus, AD biomarkers from more easily accessible body fluids, such as plasma, are very enticing. Using an aptamer-based proteomic technology, we profiled 1,129 plasma proteins of AD patients and non-demented control individuals. A 5-protein classifier for AD identification was constructed in the discovery study with excellent 10-fold cross-validation performance (90.1% sensitivity, 84.2% specificity, 87.9% accuracy, and AUC as 0.94). In an independent validation study, the classifier was applied and correctly predicted AD with 100.0% sensitivity, 80.0% specificity, and 90.0% accuracy, matching or outperforming the CSF A?42 and tau biomarkers whose performance were assessed in individual-matched CSF samples obtained at the same visit as plasma sample collection. Moreover, the classifier also correctly predicted mild cognitive impairment, an early pre-dementia state of the disease, with 96.7% sensitivity, 80.0% specificity, and 92.5% accuracy. These studies demonstrate that plasma proteins could be used effectively and accurately to contribute to the clinical diagnosis of AD. Although additional and more diverse cohorts are needed for further validation of the robustness, including the support of postmortem diagnosis, the 5-protein classifier appears to be a promising blood test to contribute diagnosis of AD.
Related JoVE Video
Redox-based selective fluorometric detection of homocysteine.
Chem. Commun. (Camb.)
PUBLISHED: 08-07-2014
Show Abstract
Hide Abstract
Plasma homocysteine (Hcy) is an important risk factor for various diseases. A novel redox-sensitive fluorescent probe is developed for the selective detection of Hcy. A linear calibration curve has been obtained in buffer and plasma for the quantitative determination of Hcy in such media.
Related JoVE Video
Synthesis of PbS/PbI2 nanocomposites in mixed solvent and their composition-dependent electrogenerated chemiluminescence performance.
Inorg Chem
PUBLISHED: 08-01-2014
Show Abstract
Hide Abstract
PbS/PbI2 nanocomposites were prepared by choosing K[PbI3] as both a lead salt and an iodide precursor and acetone/water as a reaction medium. It was found that the amount of the PbI2 component could be controlled, to some extent, by varying the amount of water used. Further, this simple bicomponent precursor-based synthetic route can be extended to prepare other lead-containing nanocomposites such as Pb3O4/PbI2 and PbSe/PbI2. Because of the heavy-atom effect, PbS/PbI2 nanocomposites exhibited good and composition-dependent electrogenerated chemiluminescence (ECL) performance, demonstrating their potential in the development of novel ECL sensors for analytical and clinical applications. These interesting findings would encourage us to gain deep insight on these phenomena, which could lead to the further development of these new inorganic materials and their applications.
Related JoVE Video
Decreased expression of prenyl diphosphate synthase subunit 2 correlates with reduced survival of patients with gastric cancer.
J. Exp. Clin. Cancer Res.
PUBLISHED: 07-27-2014
Show Abstract
Hide Abstract
BackgroundIdentification of novel molecular biomarkers will improve the management of patients with gastric cancer (GC). Prenyl diphosphate synthase subunit 2 (PDSS2) is required for coenzyme Q10 biosynthesis and acts as a tumor suppressor; however, the role and regulatory mechanisms of PDSS2 in GC are not understood. The aim of this study was to determine expression status and regulatory mechanisms of PDSS2 in GC.MethodsAssociations between expression and methylation of PDSS2 were evaluated using GC cell lines. The clinical significance of PDSS2 expression was evaluated using 238 pairs of surgically resected gastric tissues with subgroup analysis based on GC subtypes.ResultsThe expression of PDSS2 mRNA was decreased in 73% of GC cell lines compared with the control non-cancerous cell. The PDSS2 promoter was hypermethylated in cells with decreased PDSS2 expression, and treating these cells with a methylation inhibitor reactivated PDSS2 expression. GC tissues expressed significantly lower mean levels of PDSS2 mRNA compared with adjacent normal tissues (P <0.001). The expression pattern of PDSS2 protein was consistent with that of its mRNA. The decrease of PDSS2 mRNA expression in GC tissues (less than half the level of expression detected in the corresponding normal adjacent tissues) correlated significantly with elevated levels of carbohydrate antigen 19-9 (P =0.015), lymph node metastasis (P =0.022), and shorter recurrence-free survival after curative resection (P =0.022). Further, multivariate analysis identified PDSS2 mRNA expression as an independent prognostic factor (hazard ratio 1.95, 95% confidence interval 1.22¿3.09, P =0.005), and its expression pattern and prognostic significance were similar among three GC subtypes.Conclusions PDSS2 encodes a putative tumor suppressor, and we show here that its expression was regulated by hypermethylation of its promoter in GC cells. Inhibition of PDSS2 mRNA expression may serve as a novel biomarker of all types of GC.
Related JoVE Video
Stable 7,14-disubstituted-5,12-dithiapentacenes with quinoidal conjugation.
Org. Lett.
PUBLISHED: 07-14-2014
Show Abstract
Hide Abstract
Two 7,14-disubstituted-5,12-dithiapentacenes (1 and 2) with quinoidal conjugation were synthesized. Their ground-state quinoidal structures were proven by X-ray crystallographic analysis. They showed very different electronic and optical properties from those of the corresponding pentacene derivatives with diene conjugation, and their stability was significantly improved. Organic field effect transistors based on solution processed thin films of 1 and 2 exhibited a hole mobility of up to 0.032 cm(-2) V(-1) s(-1).
Related JoVE Video
Laboratory analyses of two explanted hydrophobic acrylic intraocular lenses.
Indian J Ophthalmol
PUBLISHED: 07-10-2014
Show Abstract
Hide Abstract
Two three-piece hydrophobic acrylic intraocular lenses (IOLs) were explanted from two patients at 7 and 9 years, respectively, after implantation, because of poor fundus visualisation and/or a clinically significant decrease in visual acuity related to their opacified IOLs. In addition to light microscopy, scanning electron microscopy and energy dispersive X-ray spectroscopy, confocal laser scanning microscopy was used for the first time to observe the explanted IOLs. The clinical aspect seemed to correspond to the phenomenon of surface light scattering, while laboratory analyses showed dense glistenings in the central layer of the IOL optic, which had no change next to the surface. Further studies on these phenomena are needed.
Related JoVE Video
A phosphomolybdic acid anion probe-based label-free, stable and simple electrochemical biosensing platform.
Chem. Commun. (Camb.)
PUBLISHED: 07-09-2014
Show Abstract
Hide Abstract
A versatile label-free, stable, low-cost and simple electrochemical biosensing platform has been developed based on a phosphomolybdic acid anion probe by jointly taking advantages of its native electronegativity, electrochemical activity and chemisorption with graphene oxide.
Related JoVE Video
Synthesis and characterization of electron donor-acceptor platinum(II) complexes composed of N,N-diphenylpyridineamine and triphenylamine ligands.
Inorg Chem
PUBLISHED: 06-27-2014
Show Abstract
Hide Abstract
The synthesis and electronic properties of a series of platinum(II) complexes composed of electron-donor and electron-acceptor components as potential photovoltaic materials is reported. The complexes are composed of triphenylamines (TPA) and pyridine-derivatized TPAs as the electron-donating components, and alkynyl derivatives of 2,1,3-benzothiadiazole and cyclopentadithiophenone as the electron acceptors. The complexes containing the pyridine-derivatized ligands were prepared to examine the effect that direct coordination of a heteroatom-modified TPA may have on the electronic properties of donor-acceptor (D-A) complexes. Four complexes composed of meta- and para- pyridine-derivatized TPAs were prepared, and their electronic properties were compared with three structurally similar complexes composed of TPA, as well as with purely organic D-A compounds. Data collected from UV-vis and cyclic voltammetry show minor differences on the properties of the complexes containing the pyridine-derivatized ligands when compared to the TPA analogs, exhibiting similar highest occupied molecular orbital-lowest unoccupied molecular orbital bandgaps ranging from 2.156 to 2.705 eV for the pyridine-derivatized complexes (6a,b and 7a,b), 2.038-2.320 eV for the TPA complexes (8a,b and 9a), 2.301 eV for organic molecule 10a, and 1.997 eV for 10b. All compounds are stable, exhibiting no decomposition in the solid indefinitely, and only minor decomposition in solution. All compounds were characterized by (1)H and (13)C nuclear magnetic resonance, infrared spectroscopy, and electrospray mass spectrometry. All complexes were also characterized by (31)P nuclear magnetic resonance and elemental analysis of CHN; determination of Ag content for 6a,b and 7a,b (carried through the synthetic steps) was determined by inductively coupled plasma optical emission spectrometry. The para-pyridine-derivatized complex of 2,1,3-benzothiadiazole (6a) was further characterized by X-ray crystallography as a AgNO3 clathrate. X-ray quality crystals were grown from a solution of hexanes/CH2Cl2 and from diffusion of hexanes into a CH2Cl2 solution of the complex, providing a solvent-free crystal and a solvate of CH2Cl2, respectively.
Related JoVE Video
Impact of adjuvant chemotherapy for stage I ovarian carcinoma with intraoperative tumor capsule rupture.
J. Obstet. Gynaecol. Res.
PUBLISHED: 06-06-2014
Show Abstract
Hide Abstract
Adjuvant chemotherapy is generally recommended for early stage epithelial ovarian cancer. However, it remains uncertain which histological subtypes and substages of stage I disease should receive adjuvant chemotherapy. The objective of this study is to determine the impact of chemotherapy among stage I epithelial ovarian cancer.
Related JoVE Video
CC Chemokine Receptor like 1 Functions as a Tumor Suppressor by Impairing CCR7-related Chemotaxis in Hepatocellular Carcinoma.
J. Pathol.
PUBLISHED: 06-03-2014
Show Abstract
Hide Abstract
Atypical chemokine receptors (ACRs) have been discovered to participate in the regulation of tumor behavior. Here, we reported a tumor suppressive role of a novel ACR member, CC chemokine receptor like 1 (CCRL1), in human hepatocellular carcinoma (HCC). Both mRNA and protein expressions of CCRL1 correlated with malignant phenotype of HCC cells and were significantly down-regulated in tumor tissue compared with paired normal liver tissue. In both the initial and validation cohorts (n?=?240 and 384, respectively), CCRL1 deficiency was associated with advanced tumor stage and was an independent index for worse survival and increased recurrence. Furthermore, knockdown or forced expression of CCRL1 revealed that CCRL1 suppressed the proliferation and invasion of HCC cells in vitro and reduced tumor growth and lung metastasis in vivo, with depressed levels of CCL19 and CCL21. By sequestrating CCL19 and CCL21, CCRL1 reduced their binding to CCR7 and consequently mitigated the detrimental impact of CCR7, including Akt/GSK-3? pathway activation and nuclear accumulation of ?-catenin in tumor cells. Clinically, the prognostic value of the CCR7 expression in HCC depended on the expression level of CCRL1, suggesting that CCRL1 may serve as an upstream switch for the CCR7 signaling cascade. Together, our findings suggest that CCRL1 impairs chemotactic events that associated with CCR7 in progression and metastasis of HCC. Our results also show a potential interplay between typical and atypical chemokine receptors in human cancer.
Related JoVE Video
MiR-302c inhibits tumor growth of hepatocellular carcinoma by suppressing the endothelial-mesenchymal transition of endothelial cells.
Sci Rep
PUBLISHED: 06-02-2014
Show Abstract
Hide Abstract
Endothelial cells (ECs) are critical for angiogenesis, and microRNA plays important roles in this process. In this study, we investigated the function and mechanism of miR-302c in the process of endothelial-mesenchymal transition (EndMT) in ECs. When miR-302c was overexpressed in HUVECs, the motility of the HUVECs was weakened; the expression levels of EndMT markers were also changed: vascular endothelial (VE)-cadherin was up-regulated, whereas ?-catenin, FSP1, and ?-SMA were down-regulated. Further in vivo and in vitro experiments showed that the growth of HCC was inhibited when co-cultured or co-injected with HUVECs overexpressing miR-302c. On the contrary, when miR-302c was suppressed in HUVECs, the opposite results were observed. Reporter assays showed that miR-302c inhibited metadherin (MTDH) expression through directly binding to its 3'UTR. In addition, compared to ECs isolated from normal liver tissues of HCC patients, ECs isolated from tumor tissues expressed markedly low levels of miR-302c but high levels of MTDH. These results suggest that EC-specific miR-302c suppresses tumor growth in HCC through MTDH-mediated inhibition of EndMT. MTDH and miR-302c might provide a new strategy for anti-angiogenic therapy in HCC.
Related JoVE Video
Dihydropyrimidinase-like 3 facilitates malignant behavior of gastric cancer.
J. Exp. Clin. Cancer Res.
PUBLISHED: 05-17-2014
Show Abstract
Hide Abstract
BackgroundGastric cancer (GC) remains to have a poor prognosis via diverse process of cancer progression. Dihydropyrimidinase-like 3 (DPYSL3) is a cell adhesion molecule that has been reported to be involved in the metastatic process of tumor cells. The aim of this study was to identify a novel clinically-relevant biomarker of GC.MethodsExpression analysis of DPYSL3 mRNA and protein levels was conducted using GC cell lines and 238 pairs of surgically resected gastric tissues. Correlations between expression status of DPYSL3 and clinicopathological parameters were investigated.Results DPYSL3 mRNA expression levels positively correlated with those of potentially interacting genes (VEGF, FAK and EZR) in GC cell lines. GC tissues from tumors with distant metastases (stage IV cancer) showed elevated expression levels of DPYSL3 mRNA. The DPYSL3 staining intensity in immunochemical staining was consistent with the mRNA expression patterns in GC tissues. High DPYSL3 mRNA expression in GCs was significantly associated with more malignant phenotypes and was an independent prognostic factor. Moreover, patients with high DPYSL3 mRNA expression had a significantly shorter recurrence free survival after curative resection. In subgroup analysis based on tumor histology, similar tendency was observed between patients with differentiated and undifferentiated GCs.ConclusionsExpression status of DPYSL3 in GC tissues may represent a promising biomarker for the malignant behavior of GC.
Related JoVE Video
3,6-Substituted-1,2,4,5-tetrazines: tuning reaction rates for staged labeling applications.
Org. Biomol. Chem.
PUBLISHED: 05-09-2014
Show Abstract
Hide Abstract
Cycloaddition reactions involving tetrazines have proven to be powerful bioorthogonal tools for various applications. Conceivably, sequential and selective labeling using tetrazine-based reactions can be achieved by tuning the reaction rate. By varying the substituents on tetrazines, cycloaddition rate variations of over 200 fold have been achieved with the same dienophile. Upon coupling with different dienophiles, such as norbornene, the reaction rate difference can be over 14,000 fold. These substituted tetrazines can be very useful for selective labeling under different conditions.
Related JoVE Video
Biomimetic superoxide dismutase stabilized by photopolymerization for superoxide anions biosensing and cell monitoring.
Anal. Chem.
PUBLISHED: 05-06-2014
Show Abstract
Hide Abstract
Photopolymerization strategy, as one of the immobilization methods, has attracted considerable interest because of some advantages, such as easy operation, harmlessness to the biomolecules, and long storage stability. (E)-4-(4-Formylstyryl) pyridine (formylstyrylpyridine) was prepared through Heck reaction and used as a photopolymer material to immobilize biomimetic superoxide dismutase under ultraviolet irradiation (UV) irradiation in a short time. The styrylpyridinium moiety of Formylstyrylpyridine was photoreactive and formed a dimer under UV irradiation. Mn2P2O7 multilayer sheet, a novel superoxide dismutase mimic, was synthesized. The formed photopolymer can immobilize Mn2P2O7 firmly under UV irradiation. On the basis of high catalytic activity of Mn2P2O7 biomimetic enzyme and long-term stability of Mn2P2O7-formylstyrylpyridine film, after introducing multiwalled carbon nanotubes (MWCNTs), a novel electrochemical biosensing platform called MWCNTs/Mn2P2O7-formylstyrylpyridine for superoxide anion (O2(•-)) detection was constructed. The biosensor displayed good performance for O2(•-) detection and provided a reliable platform to adhere living cells directly on the modified electrode surface. Therefore, the biosensor was successfully applied to vitro determination of O2(•-) released from living cells, which had a promising prospect for living cells monitoring and diagnosis of reactive oxygen species-related diseases.
Related JoVE Video
Genomic landscape and genetic heterogeneity in gastric adenocarcinoma revealed by whole-genome sequencing.
Nat Commun
PUBLISHED: 05-01-2014
Show Abstract
Hide Abstract
Gastric cancer (GC) is the second most common cause of cancer-related deaths. It is known to be a heterogeneous disease with several molecular and histological subtypes. Here we perform whole-genome sequencing of 49 GCs with diffuse (N=31) and intestinal (N=18) histological subtypes and identify three mutational signatures, impacting TpT, CpG and TpCp[A/T] nucleotides. The diffuse-type GCs show significantly lower clonality and smaller numbers of somatic and structural variants compared with intestinal subtype. We further divide the diffuse subtype into one with infrequent genetic changes/low clonality and another with relatively higher clonality and mutations impacting TpT dinucleotide. Notably, we discover frequent and exclusive mutations in Ephrins and SLIT/ROBO signalling pathway genes. Overall, this study delivers new insights into the mutational heterogeneity underlying distinct histologic subtypes of GC that could have important implications for future research in the diagnosis and treatment of GC.
Related JoVE Video
Accuracy of 3-T MRI using susceptibility-weighted imaging to detect meniscal tears of the knee.
Knee Surg Sports Traumatol Arthrosc
PUBLISHED: 04-22-2014
Show Abstract
Hide Abstract
To evaluate the diagnostic performance of susceptibility-weighted imaging (SWI) at 3-T magnetic resonance imaging for the assessment of meniscal tears.
Related JoVE Video
[Evaluation of signal noise ratio on analysis of clear cell renal cell carcinoma using DWI with multi-b values].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 04-16-2014
Show Abstract
Hide Abstract
To explore the influence of signal noise ratio (SNR) on analysis of clear cell renal cell carcinoma (CCRCC) using DWI with multi-b values.
Related JoVE Video
DNA origami-directed, discrete three-dimensional plasmonic tetrahedron nanoarchitectures with tailored optical chirality.
ACS Appl Mater Interfaces
PUBLISHED: 04-14-2014
Show Abstract
Hide Abstract
Discrete, three-dimensional (3D) gold nanoparticle (AuNP) tetrahedron nanoarchitectures are successfully self-assembled with DNA origami as template with high purity (>85%). A distinct plasmonic chiral response is experimentally observed from the AuNP tetrahedron nanoarchitectures and appears in a configuration-dependent manner. The chiral optical properties are then rationally engineered by modifying the structural parameters including the AuNP size and interparticle distance. Theoretical study of the AuNP tetrahedron nanoarchitectures shows the dependence of the chiral optical property on the AuNP size and interparticle distance, consistent with the ensemble averaged measurements.
Related JoVE Video
High Levels of Hepatitis B Surface Antigen are Associated with Poorer Survival and Early Recurrence of Hepatocellular Carcinoma in Patients with Low Hepatitis B Viral Loads.
Ann. Surg. Oncol.
PUBLISHED: 04-09-2014
Show Abstract
Hide Abstract
Recurrence is a disastrous outcome in patients with hepatitis-related hepatocellular carcinoma (HCC) who have undergone curative resection, and little is known about whether high levels of hepatitis B surface antigen (HBsAg) increase the risk of HCC recurrence.
Related JoVE Video
A highly immunogenic fragment derived from Zaire Ebola virus glycoprotein elicits effective neutralizing antibody.
Virus Res.
PUBLISHED: 04-03-2014
Show Abstract
Hide Abstract
In order to produce polyvalent vaccines based on single rVSV vector, we investigated the immunogenicity, antibody neutralizing activity, and antigenic determinant domain of Zaire Ebola's fragment MFL (aa 393-556) that contains furin site and internal fusion loop. Both the recombinant protein and the recombinant plasmid of fragment MFL elicited high levels of antibody, similar to those of Zaire Ebola GP (ZGP). The MFL fragment of ZGP also elicited high levels of neutralizing antibody and induced moderate cellular immune response in mice, as revealed by the proliferation and cytokine secretion of splenocytes. Through the analysis of the induction of neutralizing antibody by pVAX1-based recombinant plasmids that expressed truncated fragments of MFL, we found that the domain containing the internal fusion loop and the furin site was the major contributor of fragment MFL's immunogenicity. Furthermore, the rVSV-based bivalent vaccine expressing Sudan Ebola GP (SGP) and MFL fragment elicited efficient cross-immunity against ZGP and SGP with high levels of neutralizing antibody. Our results indicate that fragment MFL is an effective and novel antigen for the production of neutralizing antibody and polyvalent vaccines of Ebola virus.
Related JoVE Video
Tacrolimus promotes hepatocellular carcinoma and enhances CXCR4/SDF?1? expression in vivo.
Mol Med Rep
PUBLISHED: 03-12-2014
Show Abstract
Hide Abstract
The aim of our study was to elucidate the effect of tacrolimus (FK506) and of C-X-C chemokine receptor type 4 (CXCR4), which is a receptor specific to the stromal cell-derived factor-1? (SDF?1?), on growth and metastasis of hepatocellular carcinoma (HCC). Following treatment with different concentrations of FK506, AMD3100 or normal saline (NS), the proliferation of Morris rat hepatoma 3924A (MH3924A) cells was measured by the MTT assay, the expression of CXCR4 was analyzed with immunohistochemistry, and the morphological changes and the invasiveness of cells were studied with a transwell assay and under a scanning electron microscope, respectively. In addition, August Copenhagen Irish rat models implanted with tumor were used to examine the pathological changes and invasiveness of tumor in vivo, the expression of CXCR4 in tumor tissues and the expression of SDF?1? in the adjacent tissues to the HCC ones, using immunohistochemistry. In vitro, FK506 (100?1,000 µg/l) significantly promoted the proliferation of MH3924A cells (P<0.01), and increased the expression of CXCR4 in MH3924A cells, albeit with no significance (P>0.05). By contrast, AMD3100 had no effect on the proliferation of MH3924A cells, but significantly reduced the expression of CXCR4 (P<0.05). The invasiveness of MH3924A cells was significantly (P<0.01) enhanced following treatment with FK506, SDF?1?, FK506 + AMD3100, FK506 + SDF?1? or FK506 + AMD3100 + SDF?1?. In vivo, tumor weight (P=0.041), lymph node metastasis (P=0.002), the number of pulmonary nodules (P=0.012), the expression of CXCR4 in tumor tissues (P=0.048) and that of SDF?1? in adjacent tissues (P=0.026) were significantly different between the FK506-treated and the NS group. Our results suggest that FK506 promotes the proliferation of MH3924A cells and the expression of CXCR4 and SDF?1? in vivo. Therefore, inhibiting the formation of the CXCR4/SDF?1? complex may partly reduce the promoting effect of FK506 on HCC.
Related JoVE Video
Ag nanoparticle-ZnO nanowire hybrid nanostructures as enhanced and robust antimicrobial textiles via a green chemical approach.
Nanotechnology
PUBLISHED: 03-12-2014
Show Abstract
Hide Abstract
A new approach for fabrication of a long-term and recoverable antimicrobial nanostructure/textile hybrid without increasing the antimicrobial resistance is demonstrated. Using in situ synthesized Ag nanoparticles (NPs) anchored on ZnO nanowires (NWs) grown on textiles by a 'dip-in and light-irradiation' green chemical method, we obtained ZnONW@AgNP nanocomposites with small-size and uniform Ag NPs, which have shown superior performance for antibacterial applications. These new Ag/ZnO/textile antimicrobial composites can be used for wound dressings and medical textiles for topical and prophylactic antibacterial treatments, point-of-use water treatment to improve the cleanliness of water and antimicrobial air filters to prevent bioaerosols accumulating in ventilation, heating, and air-conditioning systems.
Related JoVE Video
Clinical significance of expression and epigenetic profiling of TUSC1 in gastric cancer.
J Surg Oncol
PUBLISHED: 03-10-2014
Show Abstract
Hide Abstract
The prognosis of advanced gastric cancer (GC) remains dismal. The aim of this study was to identify a novel tumor suppressor gene (TSG) with repressed transcription by aberrant DNA methylation in GC.
Related JoVE Video
Two-dimensional tin selenide nanostructures for flexible all-solid-state supercapacitors.
ACS Nano
PUBLISHED: 03-10-2014
Show Abstract
Hide Abstract
Due to their unique electronic and optoelectronic properties, tin selenide nanostructures show great promise for applications in energy storage and photovoltaic devices. Despite the great progress that has been achieved, the phase-controlled synthesis of two-dimensional (2D) tin selenide nanostructures remains a challenge, and their use in supercapacitors has not been explored. In this paper, 2D tin selenide nanostructures, including pure SnSe2 nanodisks (NDs), mixed-phase SnSe-SnSe2 NDs, and pure SnSe nanosheets (NSs), have been synthesized by reacting SnCl2 and trioctylphosphine (TOP)-Se with borane-tert-butylamine complex (BTBC) and 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone. Utilizing the interplay of TOP and BTBC and changing only the amount of BTBC, the phase-controlled synthesis of 2D tin selenide nanostructures is realized for the first time. Phase-dependent pseudocapacitive behavior is observed for the resulting 2D nanostructures. The specific capacitances of pure SnSe2 NDs (168 F g(-1)) and SnSe NSs (228 F g(-1)) are much higher than those of other reported materials (e.g., graphene-Mn3O4 nanorods and TiN mesoporous spheres); thus, these tin selenide materials were used to fabricate flexible, all-solid-state supercapacitors. Devices fabricated with these two tin selenide materials exhibited high areal capacitances, good cycling stabilities, excellent flexibilities, and desirable mechanical stabilities, which were comparable to or better than those reported recently for other solid-state devices based on graphene and 3D GeSe2 nanostructures. Additionally, the rate capability of the SnSe2 NDs device was much better than that of the SnSe NS device, indicating that SnSe2 NDs are promising active materials for use in high-performance, flexible, all-solid-state supercapacitors.
Related JoVE Video
Translocation analysis and safety assessment in two water spinach cultivars with distinctive shoot Cd and Pb concentrations.
Environ Sci Pollut Res Int
PUBLISHED: 03-02-2014
Show Abstract
Hide Abstract
A pot experiment was conducted to investigate the translocation of cadmium (Cd) and lead (Pb) and assess the safety of edible parts in two cultivars of water spinach (Ipomoea aquatica Forsk.) contrasting in shoot Cd and Pb concentrations. A low-Cd-Pb cultivar (QLQ) and a high-Cd-Pb cultivar (T308) were grown in five soils with different concentrations of Cd and Pb. The results showed that QLQ had lower Cd and Pb concentrations in stems and leaves and higher root Cd concentration than T308 did. Root Pb concentration of T308 dramatically increased with increasing soil Pb concentration and was higher than that of QLQ in the highest Pb treatment. The root-to-stem Cd translocation ability in T308 was 2.3-3.0-fold higher than that in QLQ. Nevertheless, there was no significant difference in root-to-stem Pb translocation between QLQ and T308. The bioconcentration factors (BCFs) for Cd and Pb in the two cultivars remained stable in different Cd or Pb treatments, which were attributable to the homeostatic control mechanisms of Cd and Pb in water spinach.
Related JoVE Video
Characterization of cadmium uptake, translocation, and distribution in young seedlings of two hot pepper cultivars that differ in fruit cadmium concentration.
Environ Sci Pollut Res Int
PUBLISHED: 02-20-2014
Show Abstract
Hide Abstract
The reasons why some cultivars of hot pepper (Capsicum annuum L.) accumulate low levels of Cd are poorly understood. We aimed to compare the characteristics of Cd uptake and translocation in low-Cd and high-Cd hot pepper cultivars by determining the subcellular locations and chemical forms of Cd, and its distribution among different plant organs. We conducted a hydroponic experiment to investigate the subcellular distribution and chemical forms of Cd in roots, stems, and leaves of a low-Cd (Yeshengchaotianjiao, YCT) and a high-Cd cultivar (Jinfuzaohuangjiao, JFZ). The results showed that the concentrations of Cd in almost all subcellular fractions of roots, and in all chemical forms in roots, were higher in YCT than in JFZ. Compared with YCT, JFZ had higher Cd concentrations in almost all subcellular fractions of stems and leaves, and higher Cd concentrations in almost all chemical forms in stems and leaves. Additionally, YCT had significantly higher total Cd accumulation but a lower Cd translocation rate compared with JFZ. In general, the results presented in this study revealed that root-to-shoot Cd translocation via the xylem is the key physiological processes determining the Cd accumulation level in stems and leaves of hot pepper plants. Immobilization of Cd by the cell walls of different organs is important in Cd detoxification and limiting the symplastic movement of Cd.
Related JoVE Video
Capn4 contributes to tumour growth and metastasis of hepatocellular carcinoma by activation of the FAK-Src signalling pathways.
J. Pathol.
PUBLISHED: 02-17-2014
Show Abstract
Hide Abstract
Calpain small subunit 1 (Capn4) has been identified as a major gene that promotes metastasis of hepatocellular carcinoma (HCC). However, the mechanism by which Capn4 promotes progression of HCC is not understood. In this study, we found that Capn4 expression was increased in highly metastatic HCC cell lines and in tumour tissue from HCC patients compared to healthy patient tissue. Over-expression of Capn4 in HCC cells enhanced tumour cell growth in vitro and increased invasiveness, tumourigenicity and lung metastasis in vivo. Protein microarray analyses showed that expression of multiple proteins was regulated by Capn4. Interestingly, Capn4 was found to physically associate with FAK and promoted hyperactivity of the FAK-Src signalling pathway via increased phosphorylation of specific tyrosine residues of FAK, Src and p130Cas. Knock-down of Capn4 expression suppressed the malignant behaviour of HCC cells and inhibited the FAK-Src signalling pathway. Furthermore, Capn4-mediated invasion and metastasis of HCC cells required up-regulation of matrix metalloproteinase-2 (MMP2) through activation of this signalling pathway. Our clinical data revealed that Capn4 expression correlated well with the levels of phospho-FAK, and over-expression of both Capn4 and phospho-FAK correlates with the poorest survival outcomes in HCC. In conclusion, our data showed that Capn4 can contribute to HCC growth and metastasis via activation of the FAK-Src signalling pathway and MMP2. Copyright © 2014 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Related JoVE Video
Using ruthenium polypyridyl functionalized ZnO mesocrystals and gold nanoparticle dotted graphene composite for biological recognition and electrochemiluminescence biosensing.
Nanoscale
PUBLISHED: 01-17-2014
Show Abstract
Hide Abstract
Using ruthenium polypyridyl functionalized ZnO mesocrystals as bionanolabels, a universal biological recognition and biosensing platform based on gold nanoparticle (AuNP) dotted reduced graphene oxide (rGO) composite was developed. AuNP-rGO accelerated electron transfer between the detection probe and the electrode, and increased the surface area of the working electrode to load greater amounts of the capture antibodies. The large surface area of ZnO mesocrystals was beneficial for loading a high content ruthenium polypyridyl complex, leading to an enhanced electrochemiluminescence signal. Using ?-fetoprotein (AFP) as a model, a simple and sensitive sandwich-type electrochemiluminescence biosensor with tripropylamine (TPrA) as a coreactant for detection of AFP was constructed. The designed biosensor provided a good linear range from 0.04 to 500 ng mL(-1) with a low detection limit of 0.031 ng mL(-1) at a S/N of 3 for AFP determination. The proposed biological recognition and biosensing platform extended the application of ruthenium polypyridyl functionalized ZnO mesocrystals, which provided a new promising prospect.
Related JoVE Video
Electrochemical monitoring of an important biomarker and target protein: VEGFR2 in cell lysates.
Sci Rep
PUBLISHED: 01-16-2014
Show Abstract
Hide Abstract
Vascular endothelial growth factor receptor 2 (VEGFR2) is a potential cell-type biomarker in clinical diagnoses. Besides, it's the target protein of many tyrosine kinase inhibitors and its expression significantly associates with clinical performance of these inhibitors. VEGFR2 detection provides an early warning for diseases and a basis for therapy and drug screening. Some methods have been developed for VEGFR2 determination. However, they are usually performed indirectly and complexly. Herein, an electrochemical biosensing platform for VEGFR2 analysis has been first proposed. It can detect the total concentrations of the VEGFR2 protein in cells lysates directly and can be used to monitor the changes of VEGFR2 expression levels induced by treatments of different inhibitors. Moreover, the inhibitor-VEGFR2 interactions are illuminated through theoretical simulation. The simulation results agree well with the experimental data, indicating the veracity of the proposed method. The electrochemical detection methodology for VEGFR2 would be promising in clinical diagnosis and drug screening.
Related JoVE Video
Addition of poly (propylene glycol) to multiblock copolymer to optimize siRNA delivery.
Bioengineered
PUBLISHED: 01-16-2014
Show Abstract
Hide Abstract
Previous studies have examined different strategies for siRNA delivery with varying degrees of success. These include use of viral vectors, cationic liposomes, and polymers. Several copolymers were designed and synthesized based on blocks of poly(ethylene glycol) PEG, poly(propylene glycol) PPG, and poly(l-lysine). These were designated as P1, P2, and P3. We studied the copolymer self-assembly, siRNA binding, particle size, surface potential, architecture of the complexes, and siRNA delivery. Silencing of GFP using copolymer P3 to deliver GFP-specific siRNA to Neuro-2a cells expressing GFP was almost as effective as using Lipofectamine 2000, with minimal cytotoxicity. Thus, we have provided a new copolymer platform for siRNA delivery that we can continue to modify for improved delivery of siRNA in vitro and eventually in vivo.
Related JoVE Video
In situ generated AgBr-enhanced ZnO nanorod-based photoelectrochemical aptasensing via layer-by-layer assembly.
Chem. Commun. (Camb.)
PUBLISHED: 01-13-2014
Show Abstract
Hide Abstract
A robust aptasensor for Ag(+) was proposed for the first time using an enhanced ZnO nanorod-based photoelectrochemistry by in situ generated AgBr via layer-by-layer assembly. This work opens up new avenues for application of one-dimensional ZnO nanorod arrays in photoelectrochemical sensing. Additionally, the strategy of employing in situ generated narrow-bandgap semiconductors paves a new way for photoelectrochemical sensing.
Related JoVE Video
Well-coupled graphene and Pd-based bimetallic nanocrystals nanocomposites for electrocatalytic oxygen reduction reaction.
ACS Appl Mater Interfaces
PUBLISHED: 01-13-2014
Show Abstract
Hide Abstract
In this paper, a series of well-coupled graphene (G) and MPd3 (M = Fe, Cu, Ag, Au, Cr, Mo, W) nanocrystals nanocomposites (G-MPd3 NCPs) have been synthesized via a versatile electrostatic assembly and hydrogen reduction strategy, i.e., sequential assembly of coordination anions and cations on excess cationic polymer modified graphene oxide to form composite precursors and then thermal treating under H2/Ar gases atmosphere. In those NCPs, the MPd3 components are uniform and smaller than 10 nm, which are well anchored on G with "naked" or "clean" surfaces. By adjusting reaction temperature, the interplay of MPd3 nanocrystals and G can be well-controlled. Below 700 °C, no sintering phenomena are observed, indicating the unprecedented dispersion and stability effect of G for MPd3 nanocrystals. All the obtained NCPs can be directly used to catalyze oxygen reduction reaction in alkaline media. Compared with single component, monometallic, and some reported non-Pt catalysts, greatly enhanced electrocatalytic performances are observed in those NCPs due to strong synergistic or coupling of their constituents. Among them, G-FePd3 NCPs exhibit the highest catalytic activity, but their current density needs to be improved compared with G-CrPd3, G-MoPd3, and G-WPd3 ones. This work not only provides a general strategy for fabricating well-coupled G-MPd3 NCPs but also paves the way for future designing multicomponent NCPs with multiple interfaces to apply in alkaline fuel cells.
Related JoVE Video
Dual-source parallel radiofrequency transmission for magnetic resonance breast imaging at 3T: any added clinical value?
Magn Reson Imaging
PUBLISHED: 01-12-2014
Show Abstract
Hide Abstract
To investigate the influence of dual-source parallel radiofrequency (RF) excitation on clinical breast MR images.
Related JoVE Video
Prognostic impact of expression and methylation status of DENN/MADD domain-containing protein 2D in gastric cancer.
Gastric Cancer
PUBLISHED: 01-09-2014
Show Abstract
Hide Abstract
Patients with advanced gastric cancer (GC) have an adverse prognosis even after curative resection. Development of novel diagnostic and therapeutic approaches for GC is urgently required.
Related JoVE Video
Study on the Reutilization of Clear Fracturing Flowback Fluids in Surfactant Flooding with Additives for Enhanced Oil Recovery (EOR).
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
An investigation was conducted to study the reutilization of clear fracturing flowback fluids composed of viscoelastic surfactants (VES) with additives in surfactant flooding, making the process more efficient and cost-effective. The clear fracturing flowback fluids were used as surfactant flooding system with the addition of ?-olefin sulfonate (AOS) for enhanced oil recovery (EOR). The interfacial activity, emulsification activity and oil recovery capability of the recycling system were studied. The interfacial tension (IFT) between recycling system and oil can be reduced by 2 orders of magnitude to 10-3 mN/m, which satisfies the basic demand of surfactant flooding. The oil can be emulsified and dispersed more easily due to the synergetic effect of VES and AOS. The oil-wet surface of quartz can be easily converted to water-wet through adsorption of surfactants (VES/AOS) on the surface. Thirteen core plug flooding tests were conducted to investigate the effects of AOS concentrations, slug sizes and slug types of the recycling system on the incremental oil recovery. The investigations prove that reclaiming clear fracturing flowback fluids after fracturing operation and reuse it in surfactant flooding might have less impact on environment and be more economical.
Related JoVE Video
Importance of Hyperbilirubinemia in Differentiation of Primary and Secondary Hemophagocytic Lymphohistiocytosis in Pediatric Cases.
Mediterr J Hematol Infect Dis
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening hyper-inflammatory disease. It is difficult to differentiate between primary and secondary HLH based on clinical findings at the onset of disease. We aimed to find parameters that can help to differentiate primary and secondary HLH at initial diagnosis especially for physicians working in developing countries.
Related JoVE Video
Characterization of Breast Tumors Using Diffusion Kurtosis Imaging (DKI).
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
The aim of this study was to investigate and evaluate the role of magnetic resonance (MR) diffusion kurtosis imaging (DKI) in characterizing breast lesions.
Related JoVE Video
Study of the formation and solution properties of worm-like micelles formed using both N-hexadecyl-N-methylpiperidinium bromide-based cationic surfactant and anionic surfactant.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
The viscoelastic properties of worm-like micelles formed by mixing the cationic surfactant N-hexadecyl-N-methylpiperidinium bromide (C16MDB) with the anionic surfactant sodium laurate (SL) in aqueous solutions were investigated using rheological measurements. The effects of sodium laurate and temperature on the worm-like micelles and the mechanism of the observed shear thinning phenomenon and pseudoplastic behavior were systematically investigated. Additionally, cryogenic transmission electron microscopy images further ascertained existence of entangled worm-like micelles.
Related JoVE Video
Reduced Right Frontal Fractional Anisotropy Correlated with Early Elevated Plasma LDL Levels in Obese Young Adults.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
To investigate the underlying physiological mechanisms of the structural differences in gray matter (GM) and white matter (WM) associated with obesity in young Chinese adults.
Related JoVE Video
Formation of worm-like micelles in mixed N-hexadecyl-N-methylpyrrolidinium bromide-based cationic surfactant and anionic surfactant systems.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Through the descriptive and rheological characterization of worm-like micelles formed by N-hexadecyl-N-methylpyrrolidinium bromide and sodium laurate, the formation and properties of the worm-like micelles were affected by the concentrations of sodium laurate and temperature. Additionally, cryogenic transmission electron microscopy images further validated the formation of worm-like micelles.
Related JoVE Video
Investigation of the profile control mechanisms of dispersed particle gel.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Dispersed particle gel (DPG) particles of nano- to micron- to mm-size have been prepared successfully and will be used for profile control treatment in mature oilfields. The profile control and enhanced oil recovery mechanisms of DPG particles have been investigated using core flow tests and visual simulation experiments. Core flow test results show that DPG particles can easily be injected into deep formations and can effectively plug the high permeability zones. The high profile improvement rate improves reservoir heterogeneity and diverts fluid into the low permeability zone. Both water and oil permeability were reduced when DPG particles were injected, but the disproportionate permeability reduction effect was significant. Water permeability decreases more than the oil permeability to ensure that oil flows in its own pathways and can easily be driven out. Visual simulation experiments demonstrate that DPG particles can pass directly or by deformation through porous media and enter deep formations. By retention, adsorption, trapping and bridging, DPG particles can effectively reduce the permeability of porous media in high permeability zones and divert fluid into a low permeability zone, thus improving formation profiles and enhancing oil recovery.
Related JoVE Video
Dichloroacetate enhances adriamycin-induced hepatoma cell toxicity in vitro and in vivo by increasing reactive oxygen species levels.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
A unique bioenergetic feature of cancer, aerobic glycolysis is considered an attractive therapeutic target for cancer therapy. Recently, dichloroacetate (DCA), a small-molecule metabolic modulator, was shown to reverse the glycolytic phenotype, induce reactive oxygen species (ROS) generation, and trigger apoptosis in various tumor cells. In this work, the capacity of DCA to enhance Adriamycin (ADM) efficacy in hepatoma cells by modulating glucose metabolism and redox status was evaluated. Two human hepatoma (HCC-LM3 and SMMC-7721) and a normal liver (LO2) cell lines were treated with DCA or ADM alone, or in combination. Exposure of hepatoma cells to DCA/ADM combination resulted in significantly decreased cell viability and increased percentage of apoptotic cells as well as intracellular ROS levels, in comparison with treatment with DCA or ADM alone. However, simultaneous treatment with the thiol antioxidant N-acetylcysteine (NAC, 10 mmol/L) reduced the elevated ROS levels and protected hepatoma cells from the cytotoxic effects of DCA/ADM combination. L-buthionine-[S,R]-sulfoximine, an inhibitor of glutathione synthesis, enhanced hepatoma cell sensitivity to DCA/ADM combination. Interestingly, treatment with DCA/ADM combination did not significantly increase cytotoxicity in normal hepatocytes in comparison with the drugs administered individually. Finally, DCA reduced tumor growth and enhanced ADM efficacy on HCC-LM3 hepatoma in mice. Overall, our data suggest that DCA enhances ADM cytotoxicity in hepatoma cells by increasing intracellular ROS levels and provide a strong biochemical rationale for the use of DCA in combination with ADM for treatment of hepatoma.
Related JoVE Video
Assessment of iron deposition and white matter maturation in infant brains by using enhanced T2 star weighted angiography (ESWAN): R2* versus phase values.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Iron deposition and white matter (WM) maturation are very important for brain development in infants. It has been reported that the R2* and phase values originating from the gradient-echo sequence could both reflect the iron and myelination. The aim of this study was to investigate age-related changes of R2* and phase value, and compare their performances for monitoring iron deposition and WM maturation in infant brains.
Related JoVE Video
CXCL5 contributes to tumour metastasis and recurrence of intrahepatic cholangiocarcinoma by recruiting infiltrative intratumoural neutrophils.
Carcinogenesis
PUBLISHED: 11-30-2013
Show Abstract
Hide Abstract
CXCL5 is a member of the CXC-type chemokine family that may play a role in carcinogenesis and cancer progression. This study investigates the biological function and clinical significance of CXCL5 in intrahepatic cholangiocarcinoma (ICC). We demonstrated that CXCL5 was overexpressed in ICC cell lines and tumour samples compared with paired normal tissues. CXCL5 had a direct chemoattractant effect on neutrophils in vitro through PI3K-Akt and ERK1/2 signalling pathways. In animal studies, CXCL5 promoted tumour growth and metastasis without altering in vitro proliferative and invasive ability of ICC cells, and this effect was mediated by the recruitment of intratumoural infiltrative neutrophils by tumour-derived CXCL5. Immunohistochemical analysis of ICC samples showed that overexpression of CXCL5 correlated strongly with intratumoural neutrophil infiltration, shorter overall survival, and high tumour recurrence. Multivariate analysis revealed that CXCL5 overexpression alone, or combined with the presence of intratumoural neutrophils, was an independent prognostic indicator for ICC. In conclusion, our data showed that CXCL5 promotes ICC growth and metastasis by recruiting intratumoural neutrophils. CXCL5 alone or combined with intratumoural neutrophils is a novel prognostic predictor for ICC patients and a potential therapeutic target.
Related JoVE Video
Using graphene-based plasmonic nanocomposites to quench energy from quantum dots for signal-on photoelectrochemical aptasensing.
Anal. Chem.
PUBLISHED: 11-27-2013
Show Abstract
Hide Abstract
On the basis of the absorption and emission spectra overlap, an enhanced resonance energy transfer caused by excition-plasmon resonance between reduced graphene oxide (RGO)-Au nanoparticles (AuNPs) and CdTe quantum dots (QDs) was obtained. With the synergy of AuNPs and RGO as a planelike energy acceptor, it resulted in the enhancement of energy transfer between excited CdTe QDs and RGO-AuNPs nanocomposites. Upon the novel sandwichlike structure formed via DNA hybridization, the exciton produced in CdTe QDs was annihilated. A damped photocurrent was obtained, which was acted as the background signal for the development of a universal photoelectrochemical (PEC) platform. With the use of carcinoembryonic antigen (CEA) as a model which bonded to its specific aptamer and destroyed the sandwichlike structure, the energy transfer efficiency was lowered, leading to PEC response augment. Thus a signal-on PEC aptasensor was constructed. Under 470 nm irradiation at -0.05 V, the PEC aptasensor for CEA determination exhibited a linear range from 0.001 to 2.0 ng mL(-1) with a detection limit of 0.47 pg mL(-1) at a signal-to-noise ratio of 3 and was satisfactory for clinical sample detection. Since different aptamers can specifically bind to different target molecules, the designed strategy has an expansive application for the construction of versatile PEC platforms.
Related JoVE Video
Susceptibility-weighted MR Imaging in the Grading of Liver Fibrosis: A Feasibility Study.
Radiology
PUBLISHED: 10-28-2013
Show Abstract
Hide Abstract
Purpose To assess the feasiblity of magnetic resonance (MR) susceptibility-weighted (SW) imaging as a tool to evaluate liver fibrosis grades in patients with chronic liver diseases (CLD) utilizing signal intensity (SI) measurements, with histopathologic findings as the reference standard. Materials and Methods This retrospective study was approved by the local ethics committee. All subjects gave written informed consent. Eighty consecutive patients (mean age, 56.8 years), 60% of whom were male [n = 48] and 40% of whom were female [n = 32], with CLD due to various underlying causes and histopathologically proved liver fibrosis were included. Biopsies were evaluated for liver fibrosis and necroinflammatory activity (according to METAVIR scoring system), iron load, and steatosis. Two radiologists, blinded to the clinical data, assessed regions of interest in the liver and spinal muscle in consensus. Liver-to-muscle SI ratios were calculated and correlated to histopathologic findings and clinical data by using univariate and multivariate regression analysis. Results Liver-to-muscle SI ratio decreased in parallel with the increasing grade of liver fibrosis and correlated strongly with liver fibrosis (r = -0.81, P < .0001) and moderately with necroinflammatory activity (r = -0.52, P < .0001) and iron load (r = -0.37, P = .0002) but did not correlate with steatosis (r = -0.18, P = .11). In multiple regression analysis, liver fibrosis and iron load independently influenced SW imaging measurements, explaining 69% of the variance of liver-to-muscle SI ratio (R(2) = 0.69, P < .001). Liver-to-muscle SI ratio performed well in grading liver fibrosis, with an area under the receiver operating characteristic curve of 0.92 for scores of F2 or higher and 0.93 for score of F4 (liver cirrhosis). Conclusion SW imaging is a feasible noninvasive tool to detect moderate and advanced liver fibrosis in CLD patients. © RSNA, 2013.
Related JoVE Video
Identification of intragenic methylation in the TUSC1 gene as a novel prognostic marker of hepatocellular carcinoma.
Oncol. Rep.
PUBLISHED: 10-24-2013
Show Abstract
Hide Abstract
Patients with hepatocellular carcinoma (HCC) have a poor prognosis, and novel molecular targets for treating recurrence and progression of the disease along with associated biomarkers are urgently required. In the present study, expression and the regulatory mechanism of TUSC1 (tumor suppressor candidate 1) were investigated to determine if it is a candidate tumor suppressor gene for HCC, which shows repressed transcription that involves aberrant DNA methylation. TUSC1 mRNA expression levels in HCC cell lines and 94 pairs of surgical specimens were determined using quantitative real-time reverse transcription polymerase chain reaction assay. Methylation status of HCC cell lines and clinical samples were analyzed to investigate the regulatory mechanism of TUSC1 transcription and the relationship between the methylation status of the TUSC1 gene and clinicopathological factors. The expression and distribution of the TUSC1 protein in liver tissues were determined using immunohistochemistry. A majority of HCC cell lines (89%) and surgical specimens (84%) demonstrated reduced expression levels of TUSC1 mRNA compared with paired non-cancerous liver tissues. The mean mRNA expression level in HCC was significantly lower than in corresponding non-cancerous liver. In contrast, no significant difference was found in TUSC1 mRNA expression level between adjacent normal and cirrhotic liver tissue from HCC patients. The TUSC1 protein expression pattern in HCC and liver tissues was consistent with TUSC1 mRNA expression. Twenty-nine (31%) of 94 patients showed intragenic hypermethylation of the TUSC1 gene in HCC, and hypermethylation was significantly associated with advanced pathological stage. Subsequently, patients with hypermethylation of the TUSC1 gene had a significantly poorer prognosis than patients without hypermethylation. Our results suggest that TUSC1 is a candidate tumor suppressor gene and intragenic hypermethylation is one of the suppressive mechanisms that regulate TUSC1 transcription in HCC. Intragenic methylation of the TUSC1 gene may serve as a novel prognostic marker of HCC.
Related JoVE Video
DNA-directed gold nanodimers with tailored ensemble surface-enhanced Raman scattering properties.
ACS Appl Mater Interfaces
PUBLISHED: 10-16-2013
Show Abstract
Hide Abstract
Gold nanodimers (GNDs) are assembled with high uniformity as ideal surface-enhanced Raman scattering (SERS) substrates through DNA-directed self-assembly of gold nanoparticles. The interparticle distance within GNDs is precisely tailored on the order of a few nanometers with changing the molecule length of DNA bridge. The ensemble SERS activity of monodispersed GNDs is then rationally engineered by modifying the structural parameters of GNDs including the particle size and interparticle distance. Theoretical studies on the level of single GND evidence the particle size- and interparticle-distance-dependent SERS effects, consistent with the ensemble averaged measurements.
Related JoVE Video
Autophagy inhibition suppresses pulmonary metastasis of HCC in mice via impairing anoikis resistance and colonization of HCC cells.
Autophagy
PUBLISHED: 10-01-2013
Show Abstract
Hide Abstract
Metastasis is one of the main causes of poor prognosis for hepatocellular carcinoma (HCC), which has been linked to cell-death resistance. Autophagy is an important survival mechanism under conditions of cell stress. We hypothesized that autophagy may play a role in HCC metastasis due to its prosurvival effect. Highly metastatic HCC cell lines with stable autophagy inhibition were established via lentivirus-mediated silencing of BECN1 and ATG5 genes. Mouse models of pulmonary metastasis were then developed using the cells with or without autophagy inhibition. The analysis of lung metastasis by histopathological examination and small animal imaging showed that autophagy inhibition significantly decreased the incidence of pulmonary metastases in vivo. Further invasion, migration, detachment, lung colonization, and epithelial-mesenchymal transition (EMT) assays indicated that autophagy inhibition did not affect cell invasiveness, migration or EMT but attenuated the anoikis-resistance and lung colonization of HCC cells. Investigation of the molecular mechanisms underlying showed that the autophagy-inhibition-mediated anoikis-resistance attenuation was associated with the regulation of apoptotic signaling. As autophagy inhibition was shown to be able to suppress HCC metastasis, an autophagy-based HCC tissue-specific target therapy system (AFP-Cre/LoxP-shRNA) was constructed. In vitro and in vivo analyses showed that the system was able to efficiently inhibit autophagy of HCC cells and tissue in a tissue-specific manner. Further in vivo metastasis assay showed that intratumoral administration of the system could significantly suppress lung metastasis. Together, our findings suggest that autophagy may be involved in HCC metastasis through facilitating anoikis resistance and lung colonization of HCC cells. Autophagy-based HCC tissue-specific target therapy may be a new strategy for the management of HCC metastasis.
Related JoVE Video
Margin-infiltrating CD20(+) B cells display an atypical memory phenotype and correlate with favorable prognosis in hepatocellular carcinoma.
Clin. Cancer Res.
PUBLISHED: 09-20-2013
Show Abstract
Hide Abstract
The role of infiltrating B cells in hepatocellular carcinoma has been overlooked for many years. This study is aimed to delineate the distribution, prognostic value, and functional status of B cells in human hepatocellular carcinoma.
Related JoVE Video
Reduced expression of DENND2D through promoter hypermethylation is an adverse prognostic factor in squamous cell carcinoma of the esophagus.
Oncol. Rep.
PUBLISHED: 09-14-2013
Show Abstract
Hide Abstract
Esophageal cancer ranks sixth in cancer mortality worldwide and patients with esophageal squamous cell carcinoma (ESCC) have a poor prognosis with a 5-year survival rate of less than 10%. Elucidation of the mechanisms of carcinogenesis and tumor progression in esophageal cancer is urgently required to develop targets for therapy and prognostic biomarkers. In the present study, the expression and regulatory mechanism of the differentially expressed in normal and neoplastic cells domain containing 2D (DENND2D), which is a regulator of Rab GTPases, were investigated to explore its potential as a tumor suppressor gene for ESCC. The level of DENND2D mRNA expression in ESCC cell lines and surgical specimens was determined using a quantitative real-time reverse transcription-polymerase chain reaction assay, and the relationship between the expression levels of DENND2D mRNA and clinicopathological factors was evaluated. The expression and distribution of DENND2D were determined using immunohistochemistry. DNA methylation analysis was performed to determine the regulatory mechanism of DENND2D expression in ESCC. The level of DENND2D mRNA expression was reduced in 8/9 ESCC cell lines and in 59/65 surgical specimens, and the mean expression levels were significantly lower in cancerous tissues compared to corresponding normal tissues (p<0.001). The expression pattern of DENND2D protein and mRNA was consistent. Downregulation of DENND2D mRNA in ESCC tissues was identified as an independent prognostic factor in multivariate analysis (hazard ratio, 2.194; p=0.039). The DENND2D promoter was methylated in 5/9 ESCC cell lines, and DNA demethylation reactivated DENND2D mRNA expression. Hypermethylation of DENND2D was frequently detected in ESCC tissues (64.6%) and was significantly associated with downregulation of DENND2D mRNA expression (P=0.008). Taken together, our data suggest that DENND2D is a candidate tumor suppressor gene that was inactivated by promoter hypermethylation in patients with ESCC and may serve as a novel biomarker of ESCC.
Related JoVE Video

What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

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