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Find video protocols related to scientific articles indexed in Pubmed.
[Myocardial protective effect of Danhong Injection evaluated by velocity vector imaging in patients undergoing percutaneous coronary intervention].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 11-18-2014
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To observe the myocardial protective effect of Danhong Injection evaluated by velocity vector imaging (VVI) in patients with unstable angina pectoris after percutaneous coronary intervention (PCI) and elucidate its possible mechanism.
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A highly efficient and eco-friendly method for the synthesis of 1,3-indandione ring-fused 3-oxindoles bearing two contiguous quaternary stereocenters via an aldol reaction in aqueous media.
Org. Biomol. Chem.
PUBLISHED: 11-11-2014
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A highly efficient and environmentally benign method for the synthesis of oxindoles featuring two contiguous quaternary carbon centers via an aldol reaction starting from various 3-substituted oxindoles has been established. A wide variety of such featured multi-substituted 1,3-indandione ring-fused 3-oxindole scaffolds were obtained smoothly in good yields (up to 98%) employing the most green of solvents, namely water, as reaction medium. Furthermore, their biological activity has been preliminarily demonstrated by in vitro evaluation against human prostate cancer cells PC-3, human lung cancer cells A549 and human leukemia cells K562 by MTT-based assays, using the commercially available standard drug, cisplatin, as a positive control. Gratifyingly, compounds , , and exhibited the best levels of in vitro inhibitory activity against human leukemia cells K562, which were almost 2.0, 2.8, 2.5 and 2.2 times, respectively, the activity of the positive control, cisplatin. Compound had 2.7 times the activity of the positive control, cisplatin, against PC-3 cancer cells, and , and showed levels of in vitro inhibitory activity against PC-3 cancer cells that were comparable to that of cisplatin. Compounds , and had good inhibitory ability against human lung cancer cells A549. The results indicated that 1,3-indandione ring-fused 3-oxindole analogs may be useful leads for further biological screenings.
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Electrogenerated Chemiluminescence Resonance Energy Transfer between Luminol and CdSe@ZnS Quantum Dots and Its Sensing Application in the Determination of Thrombin.
Anal. Chem.
PUBLISHED: 11-07-2014
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In this work, electrogenerated chemiluminescence resonance energy transfer (ECL-RET) between luminol as a donor and CdSe@ZnS quantum dots (QDs) as an acceptor was reported in neutral conditions. It was observed that a glassy carbon electrode modified with CdSe@ZnS quantum dots (CdSe@ZnS/GCE) can catalyze the luminol oxidation to promote the anodic luminol ECL without coreactants. The intensity of anodic luminol ECL (0.60 V) at the CdSe@ZnS/GCE was enhanced more than 1 order of magnitude compared with that at the bare GCE. Another stronger anodic ECL peak observed at more positive potential (1.10 V) could be assigned to the ECL-RET between the excited state of luminol and the QDs. A label-free ECL aptasensor for the detection of thrombin was fabricated based on the synergic effect of the electrocatalysis and the ECL-RET. The approach showed high sensitivity, good selectivity, and wide linearity for the detection of thrombin in the range of 10 fM-100 pM with a detection limit of 1.4 fM (S/N = 3). The results suggested that the as-proposed luminol-QDs ECL biosensor will be promising in the detection of protein.
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Baseline Factors Associated with Mortality within Six Months after Admission among Hospitalized HIV-1 Patients in Shenyang, China.
Intern. Med.
PUBLISHED: 11-01-2014
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Objective Short-term mortality rates remain high among critically ill human immunodeficiency virus-1 (HIV-1) patients though long-term mortality rates have dropped. Baseline risk factors for short-term mortality have not yet been determined in China. In this paper, we herein describe clinical characteristics, laboratory findings, causes of clinical deterioration, and risk factors associated with mortality among HIV-1 patients within six months after hospital admission. Methods We carried out a prospective study of hospitalized patients in advanced stages of HIV infection. These patients started antiretroviral therapy three or four weeks after admission. Follow-up was conducted for a period of six months. We used a multivariate logistic-regression analysis to identify risk factors associated with mortality. Results A total of 141 patients met our inclusion criteria. The mean age was 41 years. Fever and weight loss were the most common clinical manifestations of advanced HIV disease. Oral candidiasis, tuberculosis, cytomegaloviremia, and pneumocystis pneumonia were the most common opportunistic infections. Significantly decreased CD4+ T-cell counts, hypoalbuminemia, anemia, hyponatremia, as well as elevated C-reactive protein (CRP) and glutamic alanine transaminase levels were common laboratory test abnormalities. The mortality rate was 21.3%. The patients who died were more likely than the survivors to have low CD4+ T-cell counts as well as low creatinine, hemoglobin, albumin, and serum sodium levels while also having longer intervals of fever and higher CRP levels. A multivariate analysis demonstrated that the independent risk factors for mortality were active tuberculosis [odds ratio (OR): 2.681; 95% confidence interval (CI), 1.006-7.142; p=0.049], hyponatremia (OR: 3.027; 95% CI, 1.238-7.401; p=0.015), and being at clinical stage 4 (as defined by the World Health Organization) (OR: 9.492; 95% CI, 1.200-75.065; p=0.033) within the first six months of admission. Conclusion Special consideration should be given to patients who have active tuberculosis, are at clinical stage 4, and present with hyponatremia upon admission as these were found to be important factors associated with mortality within six months of hospital admission in HIV-1 patients.
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The Identification of a Novel HIV-1 CRF01_AE/B Recombinant Using the Near Full Length Genome in Jiangsu Province, China.
AIDS Res. Hum. Retroviruses
PUBLISHED: 10-28-2014
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Abstract CRF01_AE and subtype B are the two major HIV-1 clades circulating in China. Heterosexual transmission is the predominant route for the spread of HIV and heterosexuals often include men who have sex with men and intravenous drug users. Furthermore, many kinds of circulating recombinant forms (CRF) and unique recombinant forms (URF) between CRF01_AE and subtype B were recently identified in Southeast Asia. Therefore it is inevitable that the new recombinant of CRF01_AE/B will emerge among them. Here we identified a novel recombinant of CRF01_AE/B, isolated from heterosexuals, which has a distinctly different genome structure from other CRF01Bs and URFs reported before. The analysis of the near full-length sequence of JS2011001 shows that it is composed of at least five interlaced CRF01_AE and B segments. Recently, many kinds of URFs and CRFs began to prevail within a short period in China, which implies that a mix of HIV-1 infections is common in China and more attention should focus on it.
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Ultrasensitive immunoassay based on a pseudobienzyme amplifying system of choline oxidase and luminol-reduced Pt@Au hybrid nanoflowers.
Chem. Commun. (Camb.)
PUBLISHED: 10-15-2014
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A multi-functional luminol-reduced Pt@Au hybrid flower-like nanocomposite (luminol-Pt@AuNF) which not only acts as an efficient signal probe but also constitutes a pseudobienzyme amplifying system with choline oxidase (ChOx) was firstly synthesized and applied to the construction of a solid-state luminol electrochemiluminescence (ECL) immunosensor for cardiac troponin I (cTnI) detection.
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The Effect of Heavy Metal-Induced Oxidative Stress on the Enzymes in White Rot Fungus Phanerochaete chrysosporium.
Appl. Biochem. Biotechnol.
PUBLISHED: 10-14-2014
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Prevalence of heavy metals in the living environment causes chemical stress and reactive oxygen species (ROS) formation in Phanerochaete chrysosporium (P. chrysosporium). However, the mechanisms involved in ROS defense are still under investigation. In the present study, we evaluated the effect of lead- and cadmium-induced oxidative stress on the activities of catalase (CAT), peroxidase (POD), lignin peroxidase (LiP), and manganese peroxidase (MnP). A time-dependent change in all enzyme activities was observed following exposure to 50 ?M cadmium and 25 ?M lead. The lowest values were recorded at 4 h after exposure. Both cadmium and lead inhibited CAT and POD. The cytochrome P450 (CYP450) levels increased under 50-100 ?M cadmium or lead exposure and decreased when heavy metal concentration was under 50 ?M; this suggested that ROS is not the only factor that alters the CYP450 levels. The cadmium removal rate in the sample containing 900 ?M taxifolin (inhibitor of CYP450) and 100 ?M cadmium was reduced to 12.34 %, 9.73 % lower than that of 100 ?M cadmium-induced sample, indicating CYP450 may play an indirect but key role in the process of clearance of heavy metals. The pH of the substrate solution decreased steadily during the incubation process.
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Highly regioselective synthesis of 3-alkenyl-oxindole ring-fused 3,3'-disubstituted oxindoles via direct gamma-substitution of Morita-Baylis-Hillman carbonates of isatins with 3-substituted oxindoles.
Org. Biomol. Chem.
PUBLISHED: 10-14-2014
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The first phase transfer-catalysed direct ?-substitution of Morita-Baylis-Hillman carbonates of isatins with 3-substituted oxindoles has been developed, which affords 3-alkenyl-oxindole ring-fused 3,3'-disubstituted oxindoles in up to 83% yield under mild reaction conditions. Furthermore, their biological activity has been preliminarily demonstrated by in vitro evaluation against human prostate cancer cells PC-3 and human leukemia cells K562, using MTT-based assays with the commercially available standard drug Cisplatin as a positive control. Gratifyingly, compounds 3aa, 3ba and 3ca exhibited comparable in vitro inhibitory activities against human prostate cancer cells (PC-3) to Cisplatin. What's more, 3ba also had a good inhibition ability against human leukemia cells K562. These results indicate that 3-alkenyl-oxindole ring-fused 3,3'-disubstituted oxindole analogs may be potential lead compounds for further biological screening.
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Protein synthesis using a reconstituted cell-free system.
Curr Protoc Mol Biol
PUBLISHED: 10-02-2014
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Most cell-free protein-synthesis systems are based on cell extracts, which often contain undesirable activities. Reconstituted systems, by contrast, are composed of a defined number of purified and recombinant components with minimal nuclease and protease activities. This unit describes the use of a particular commercial reconstituted system, PURExpress. This system allows in vitro synthesis of proteins from mRNA and circular and linear DNA templates, as well as co-translational labeling of proteins. Unique to this system, all recombinant protein components of the system are His-tagged, allowing purification of the synthesized untagged protein by removing the rest of the system's components. Newly synthesized proteins can often be visible on an SDS-PAGE gel and directly assayed for their functions without labeling and purification. Certain components of the system, such as ribosomes or release factors, can be omitted for specific applications. Such "delta" versions of the system are well suited for studies of bacterial translation, assays of ribosome function, incorporation of unnatural amino acids, and ribosome display of protein libraries. Curr. Protoc. Mol. Biol. 108:16.31.1-16.31.22. © 2014 by John Wiley & Sons, Inc.
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A lysosome-targeted fluorescent chemodosimeter for monitoring endogenous and exogenous hydrogen sulfide by in vivo imaging.
Chem. Commun. (Camb.)
PUBLISHED: 09-26-2014
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A lysosome-targeted fluorescent chemodosimeter, , was developed for monitoring endogenous and exogenous H2S by in vivo imaging of HeLa cells, D. melanogaster and C. elegans. In the tests of mutated C. elegans (SRP-6 nulls), chemodosimeter could trace the accumulation of lysosome and lysosomal injury with a high resolution.
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Engineering Bacterial Transcription Regulation To Create a Synthetic in Vitro Two-Hybrid System for Protein Interaction Assays.
J. Am. Chem. Soc.
PUBLISHED: 09-26-2014
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Transcriptional activation of ?(54)-RNA polymerase holoenzyme (?(54)-RNAP) in bacteria is dependent on a cis-acting DNA element (bacterial enhancer), which recruits the bacterial enhancer-binding protein to contact the holoenzyme via DNA looping. Using a constructive synthetic biology approach, we recapitulated such process of transcriptional activation by recruitment in a reconstituted cell-free system, assembled entirely from a defined number of purified components. We further engineered the bacterial enhancer-binding protein PspF to create an in vitro two-hybrid system (IVT2H), capable of carrying out gene regulation in response to expressed protein interactions. Compared with genetic systems and other in vitro methods, IVT2H not only allows detection of different types of protein interactions in just a few hours without involving cells but also provides a general correlation of the relative binding strength of the protein interaction with the IVT2H signal. Due to its reconstituted nature, IVT2H provides a biochemical assay platform with a clean and defined background. We demonstrated the proof-of-concept of using IVT2H as an alternative assay for high throughput screening of small-molecule inhibitors of protein-protein interaction.
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Studies on the effect of thymine-mercury-thymine stem as a structural or functional motif in DNAzymes.
Nucleosides Nucleotides Nucleic Acids
PUBLISHED: 09-16-2014
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T-Hg-T base pair formation has been demonstrated to be compatible with duplex DNA context, with considerable thermal stability contribution. Here, the T-Hg-T stem in two small DNAzymes 8-17 and 10-23 was studied for its structural and functional roles. The recognition arm 5' to the cleavage site of 10-23 DNAzyme complex and the stem in the catalytic loop of 8-17 DNAzyme could be replaced by consecutive T-Hg-T stem of different length. The linear relationship between the activity of the complex 10-23DZ-6T+D19-6T and the concentration of Hg(2+) demonstrated that the T-Hg-T stem contributes thermal stability of the recognition arm binding. The effect of T-Hg-T stem in the catalytic core of 8-17 DNAzyme and the position-dependent effect in 10-23 DNAzyme demonstrated that T-Hg-T base pair is not compatible with canonical base pairs in playing the functions of nucleic acids.
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[Growth activity of osteoblast on a novel strontium incorporated calcium sulfate].
Zhongguo Gu Shang
PUBLISHED: 08-30-2014
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To investigate the growth activity of osteoblast on a novel strontium incorporated calcium sulfate and make comparison with normal calcium sulfate material.
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Identification of Annexin A4 as a hepatopancreas factor involved in liver cell survival.
Dev. Biol.
PUBLISHED: 08-29-2014
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To gain insight into liver and pancreas development, we investigated the target of 2F11, a monoclonal antibody of unknown antigen, widely used in zebrafish studies for labeling hepatopancreatic ducts. Utilizing mass spectrometry and in vivo assays, we determined the molecular target of 2F11 to be Annexin A4 (Anxa4), a calcium binding protein. We further found that in both zebrafish and mouse endoderm, Anxa4 is broadly expressed in the developing liver and pancreas, and later becomes more restricted to the hepatopancreatic ducts and pancreatic islets, including the insulin producing ß-cells. Although Anxa4 is a known target of several monogenic diabetes genes and its elevated expression is associated with chemoresistance in malignancy, its in vivo role is largely unexplored. Knockdown of Anxa4 in zebrafish leads to elevated expression of caspase 8 and ?113p53, and liver bud specific activation of Caspase 3 and apoptosis. Mosaic knockdown reveal that Anxa4 is required cell-autonomously in the liver bud for cell survival. This finding is further corroborated with mosaic anxa4 knockout studies using the CRISPR/Cas9 system. Collectively, we identify Anxa4 as a new, evolutionarily conserved hepatopancreatic factor that is required in zebrafish for liver progenitor viability, through inhibition of the extrinsic apoptotic pathway. A role for Anxa4 in cell survival may have implications for the mechanism of diabetic ß-cell apoptosis and cancer cell chemoresistance.
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New organic donor-acceptor-?-acceptor sensitizers for efficient dye-sensitized solar cells and photocatalytic hydrogen evolution under visible-light irradiation.
ChemSusChem
PUBLISHED: 08-25-2014
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Two organic donor-acceptor-?-acceptor (D-A-?-A) sensitizers (AQ and AP), containing quinoxaline/pyrido[3,4-b]pyrazine as the auxiliary acceptor, have been. Through fine-tuning of the auxiliary acceptor, a higher designed and synthesized photoelectric conversion efficiency of 6.02% for the AQ-based dye-sensitized solar cells under standard global AM1.5 solar conditions was achieved. Also, it was found that AQ-Pt/TiO2 photocatalysts displayed a better rate of H2 evolution under visible-light irradiation (420 nm
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[Effects of mild hypothermia plus ifenprodil on apoptosis inducing factor translocation after global cerebral ischemia-reperfusion in rats].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 08-22-2014
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To explore the effects of mild hypothermia combined with ifenprodil on the survival of neuronal and translocation of apoptosis inducing factor (AIF) following global cerebral ischemia-reperfusion to understand the mechanism of combination in cerebral resuscitation.
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Simultaneous determination of clevidipine and its primary metabolite in dog plasma by liquid chromatography-tandem mass spectrometry: Application to pharmacokinetic study.
J Pharm Biomed Anal
PUBLISHED: 08-17-2014
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A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine and its primary metabolite H152/81 in dog plasma after protein precipitation with acetonitrile using felodipine as the internal standard (IS). Chromatographic separation was performed on a XB C18 column (2.1mm×50mm, 3.5?m) under isocratic conditions with the mobile phase consisting of acetonitrile and 20mM ammonium acetate buffer (pH 7.0) (40:60, v/v) at the flow rate of 0.3ml/min. The run time was 5.5min. Mass spectrometric analysis was performed on a triple quadrupole mass spectrometer operated in the multiple reaction monitoring (MRM) mode with the transitions of m/z 473.0?338.2 for clevidipine, m/z 356.1?324.0 for H152/81 and m/z 383.9?338.2 for the IS. The method was fully validated in terms of selectivity, linearity, lower limit of quantification (LLOQ), accuracy, precision, stability, matrix effect and recovery over a concentration range of 0.15-200ng/ml for clevidipine and 10-2000ng/ml for H152/81, respectively. The analytical method was applied to support a pharmacokinetic study of simultaneous determination of clevidipine and H152/81 in ten healthy beagle dogs.
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A time course-dependent metastatic gene expression signature predicts outcome in human metastatic melanomas.
Diagn Pathol
PUBLISHED: 08-13-2014
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The prognosis of patients with metastatic melanomas is extremely heterogeneous. Therefore, identifying high-risk subgroups by using innovative prediction models would help to improve selection of appropriate management options.
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A new method for modeling coalescent processes with recombination.
BMC Bioinformatics
PUBLISHED: 08-11-2014
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Recombination plays an important role in the maintenance of genetic diversity in many types of organisms, especially diploid eukaryotes. Recombination can be studied and used to map diseases. However, recombination adds a great deal of complexity to the genetic information. This renders estimation of evolutionary parameters more difficult. After the coalescent process was formulated, models capable of describing recombination using graphs, such as ancestral recombination graphs (ARG) were also developed. There are two typical models based on which to simulate ARG: back-in-time model such as ms and spatial model including Wiuf&Hein's, SMC, SMC', and MaCS.
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The expression of epithelial-mesenchymal transition-related proteins in biliary epithelial cells is associated with liver fibrosis in biliary atresia.
Pediatr. Res.
PUBLISHED: 08-05-2014
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BackgroundThe epithelial-mesenchymal transition (EMT) has been implicated as a key mechanism in the pathogenesis of liver fibrosis. The miR-200 family has been shown to inhibit EMT.MethodsLiver fibrosis levels were assessed with Masson's trichrome staining of liver samples obtained from biliary atresia (BA) patients.The expressions of cytokeratin-7 (CK-7) and alpha-smooth muscle actin (a-SMA) in the liver sections were detected by immunohistochemical and immunofluorescent staining. EMTs were induced by transforming growth factor (TGF)-?1 in human biliary epithelial cells (BECs) in vitro.ResultsWe showed that the EMT-related proteins CK-7 and a-SMA co-localised to the intrahepatic BECs in the liver sections of patients with BA. The level of a-SMA expression was related to liver fibrosis stage in BA. EMT in primary human intrahepatic BECs was induced by TGF-?1 in vitro. miR-200b is one member of the miR-200 family and significantly inhibited TGF-?1-mediated EMT in BECs.ConclusionTogether, these data suggest that the occurrence of EMT in BECs might contribute to BA fibrosis. miR-200b significantly affects the development and progression of TGF-?1-dependent EMT and fibrosis in vitro.Pediatric Research (2014); doi:10.1038/pr.2014.181.
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A computational study on the photoelectric properties of various Bi2O 3 polymorphs as visible-light driven photocatalysts.
J Mol Model
PUBLISHED: 08-03-2014
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This paper presents first-principle studies on the photoelectric properties of various Bi2O3 polymorphs. The intrinsic reason of different photocatalytic activities was revealed by electronic structures and optical features. Results showed that for ?, ?, and ?-Bi2O3, the top of valence bands were mainly constructed by Bi6s and O2p orbitals, and the bottom of conduction bands were dominantly composed by Bi6p orbital. However, two intermediate bands were found at the Fermi level for ?-Bi2O3, which leads to a two-step transition from the top of valence band to the bottom of conduction band and facilitates electron transition under irradiation. Absent forbidden gap was found in ?-Bi2O3, resulting in a semimetallic character due to its intrinsic oxygen vacancy and high ionic conductivity. Moreover, the optical properties of ?, ?, and ?-Bi2O3 were investigated by absorption spectrum, dielectric constant function, and energy loss spectroscopy. We concluded that the photocatalytic activities followed in the order of ?-Bi2O3?>??-Bi2O3?>??-Bi2O3, in accord with the experimental report. Calculation results illustrated the experimental observations and provided a useful guidance in exploring promising visible-light semiconductor photocatalysts.
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An agent-based model of the population dynamics of Anopheles gambiae.
Malar. J.
PUBLISHED: 08-01-2014
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Agent-based models (ABMs) have been used to model the behaviour of individual mosquitoes and other aspects of malaria. In this paper, a conceptual entomological model of the population dynamics of Anopheles gambiae and the agent-based implementations derived from it are described. Hypothetical vector control interventions (HVCIs) are implemented to target specific activities in the mosquito life cycle, and their impacts are evaluated.
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Occurrence of glycosidically conjugated 1-phenylethanol and its hydrolase ?-primeverosidase in tea (Camellia sinensis) flowers.
J. Agric. Food Chem.
PUBLISHED: 08-01-2014
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A previous study found that 1-phenylethanol (1PE) was a major endogenous volatile compound in tea (Camellia sinensis) flowers and can be transformed to glycosically conjugated 1PE (1PE-Gly). However, occurrences of 1PE-Gly in plants remain unknown. In this study, four 1PE-Glys have been isolated from tea flowers. Three of them were determined as (R)-1PE ?-d-glucopyranoside ((R)-1PE-Glu), (S)-1PE-Glu, and (S)-1PE ?-primeveroside ((S)-1PE-Pri), respectively, on the basis of NMR, MS, LC-MS, and GC-MS evidence. The other one was identified as (R)-1PE-Pri on the basis of LC-MS and GC-MS data. Moreover, these 1PE-Glys were chemically synthesized as the authentic standards to further confirm their occurrences in tea flowers. 1PE-Glu had a higher molar concentration than 1PE-Pri in each floral stage and organ. The ratio of (R) to (S) differed between 1PE-Glu and 1PE-Pri. In addition, a 1PE-Gly hydrolase ?-primeverosidase recombinant protein produced in Escherichia coli exhibited high hydrolysis activity toward (R)-1PE-Pri. However, ?-primeverosidase transcript level was not highly expressed in the anther part, which accumulated the highest contents of 1PE-Gly and 1PE. This suggests that 1PE-Gly may not be easily hydrolyzed to liberate 1PE in tea flowers. This study provides evidence of occurrences of 1PE-Glys in plants for the first time.
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PGMS: a case study of collecting PDA-based geo-tagged malaria-related survey data.
Am. J. Trop. Med. Hyg.
PUBLISHED: 07-21-2014
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Using mobile devices, such as personal digital assistants (PDAs), smartphones, tablet computers, etc., to electronically collect malaria-related field data is the way for the field questionnaires in the future. This case study seeks to design a generic survey framework PDA-based geo-tagged malaria-related data collection tool (PGMS) that can be used not only for large-scale community-level geo-tagged electronic malaria-related surveys, but also for a wide variety of electronic data collections of other infectious diseases. The framework includes two parts: the database designed for subsequent cross-sectional data analysis and the customized programs for the six study sites (two in Kenya, three in Indonesia, and one in Tanzania). In addition to the framework development, we also present our methods used when configuring and deploying the PDAs to 1) reduce data entry errors, 2) conserve battery power, 3) field install the programs onto dozens of handheld devices, 4) translate electronic questionnaires into local languages, 5) prevent data loss, and 6) transfer data from PDAs to computers for future analysis and storage. Since 2008, PGMS has successfully accomplished quite a few surveys that recorded 10,871 compounds and households, 52,126 persons, and 17,100 bed nets from the six sites. These numbers are still growing.
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Streptosporangium subfuscum sp. nov., isolated from the rhizosphere of marigold (Tagetes erecta L.).
Antonie Van Leeuwenhoek
PUBLISHED: 07-06-2014
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A Gram-stain positive, filamentous bacterial strain, designated strain NEAU-TWSJ13(T), was isolated from the rhizosphere of a marigold (Tagetes erecta L.) plant collected in Heilongjiang Province, northeast China, and characterized using a polyphasic approach. The strain was observed to form abundant aerial hyphae differentiated into spherical sporangia. 16S rRNA gene sequence similarity studies showed that strain NEAU-TWSJ13(T) belongs to the genus Streptosporangium, being most closely related to Streptosporangium fragile DSM 43847(T) (98.6 %). Phylogenetic analysis of the 16S rRNA gene sequence indicated that it formed a phyletic line with S. fragile DSM 43847(T), Streptosporangium jomthongense NBRC 110047(T) (98.4 % 16S rRNA gene similarity) and Streptosporangium violaceochromogenes DSM 43849(T) (97.6 % 16S rRNA gene similarity). A combination of DNA-DNA hybridization results and some phenotypic characteristics indicated that strain NEAU-TWSJ13(T) can be distinguished from S. fragile DSM 43847(T) and S. jomthongense NBRC 110047(T). Moreover, strain NEAU-TWSJ13(T) can also be differentiated from S. violaceochromogenes DSM 43849(T) and other Streptosporangium species showing high 16S rRNA gene sequence similarity (>98.0 %) by morphological and physiological characteristics. Therefore, it is proposed that strain NEAU-TWSJ13(T) represents a novel species of the genus Streptosporangium, for which the name Streptosporangium subfuscum sp. nov. is proposed. The type strain is NEAU-TWSJ13(T) ( = CGMCC 4.7146(T) = DSM = 46724(T)).
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Sphaerisporangium dianthi sp. nov., an endophytic actinomycete isolated from a root of Dianthus chinensis L.
Antonie Van Leeuwenhoek
PUBLISHED: 07-04-2014
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A novel actinomycete, designated strain NEAU-CY18(T), was isolated from the root of a Chinese medicinal plant Dianthus chinensis L and subjected to a polyphasic taxonomic study. The novel strain was found to develop spherical sporangia with non-motile spores on aerial mycelium. The cell-wall peptidoglycan was found to contain meso-diaminopimelic acid. The whole-cell sugars were identified as madurose, mannose, ribose, galactose and glucose. The phospholipid profile was found to contain diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, hydroxy-phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and an unidentified phospholipid. The predominant menaquinones were identified as MK-9(H4), MK-9(H2) and MK-9(H6). The major fatty acids were identified as C17:0 10-methyl, iso-C16:0 and C16:0. EzTaxon-e analysis of the 16S rRNA gene sequence indicated that the strain belongs to the genus Sphaerisporangium and was most closely related to Sphaerisporangium cinnabarinum JCM 3291(T) (98.9 %) and Sphaerisporangium melleum JCM 13064(T) (98.3 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-CY18(T) forms a monophyletic clade with S. cinnabarinum JCM 3291(T), an association that was supported by a bootstrap value of 97 % in the neighbour-joining tree and also recovered with the maximum-likelihood algorithm. Comparisons of some phenotypic properties and low DNA-DNA relatedness values enabled the strain to be differentiated from S. cinnabarinum JCM 3291(T) and S. melleum JCM 13064(T). Therefore, it is concluded that strain NEAU-CY18(T) represents a novel Sphaerisporangium species, for which the name Sphaerisporangium dianthi sp. nov. is proposed. The type strain is NEAU-CY18(T) ( = CGMCC 4.7132(T) = DSM 46736(T)).
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Designing and fabricating double resonance substrate with metallic nanoparticles-metallic grating coupling system for highly intensified surface-enhanced Raman spectroscopy.
Analyst
PUBLISHED: 07-01-2014
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Recently, nanoparticle-film coupling systems in which metal nanoparticles (supported localized surface plasmons, LSPs) are separated from a flat metal film (supported surface plasmon polaritons, SPPs) by a spacer have been widely reported due to its strong local enhancement field. However, there is are limited studies, which employ the design of combing metal grating into the nanoparticle-film gap system. Here, we propose and fabricate a novel double-resonance SERS system by strategically assembling Au NPs separated by a MoO3 nanospacer from an Ag grating film. The Ag grating with clear SPP effect is used for the first time in a double-resonance system, and the monolayer Au NPs array is well assembled onto the top of the Ag grating with a compact and uniform distribution (inter-particles gap of about 5 nm). As a result, we experimentally and theoretically demonstrate a significant near-field enhancement. The very strong near-field produced in the proposed SERS substrates is due to multiple couplings, including the Au NPs-Ag grating film coupling and Au NPs-Au NPs coupling. In addition, the as-proposed SERS substrates show good reproducibility of SERS, which have potential applications in plasmonic sensing and analytical science.
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Dissecting and engineering of the TetR family regulator SACE_7301 for enhanced erythromycin production in Saccharopolyspora erythraea.
Microb. Cell Fact.
PUBLISHED: 06-29-2014
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Background Saccharopolyspora erythraea was extensively utilized for the industrial-scale production of erythromycin A (Er-A), a macrolide antibiotic commonly used in human medicine. Yet, S. erythraea lacks regulatory genes in the erythromycin biosynthetic gene (ery) cluster, hampering efforts to enhance Er-A production via the engineering of regulatory genes.ResultsBy the chromosome gene inactivation technique based on homologous recombination with linearized DNA fragments, we have inactivated a number of candidate TetR family transcriptional regulators (TFRs) and identified one TFR (SACE_7301) positively controlling erythromycin biosynthesis in S. erythraea A226. qRT-PCR and EMSA analyses demonstrated that SACE_7301 activated the transcription of erythromycin biosynthetic gene eryAI and the resistance gene ermE by interacting with their promoter regions with low affinities, similar to BldD (SACE_2077) previously identified to regulate erythromycin biosynthesis and morphological differentiation. Therefore, we designed a strategy for overexpressing SACE_7301 with 1 to 3 extra copies under the control of PermE* in A226. Following up-regulated transcriptional expression of SACE_7301, eryAI and ermE, the SACE_7301-overexpressed strains all increased Er-A production over A226 proportional to the number of copies. Likewise, when SACE_7301 was overexpressed in an industrial S. erythraea WB strain, Er-A yields of the mutants WB/7301, WB/2×7301 and WB/3×7301 were respectively increased by 17%, 29% and 42% relative to that of WB. In a 5 L fermentor, Er-A accumulation increased to 4,230 mg/L with the highest-yield strain WB/3×7301, an approximately 27% production improvement over WB (3,322 mg/L).ConclusionsWe have identified and characterized a TetR family transcriptional regulator, SACE_7301, in S. erythraea that positively regulated erythromycin biosynthesis, and overexpression of SACE_7301 in wild-type and industrial S. erythraea strains enhanced Er-A yields. This study markedly improves our understanding of the unusual regulatory mechanism of erythromycin biosynthesis, and provides a novel strategy towards Er-A overproduction by engineering transcriptional regulators of S. erythraea.
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An update on overweight and obesity in rural Northeast China: from lifestyle risk factors to cardiometabolic comorbidities.
BMC Public Health
PUBLISHED: 06-28-2014
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Not enough is known about the prevalence of overweight and obesity in rural China in the current decade. We aim to update our knowledge of the prevalence of obesity and its associated risk factors and comorbidities in a large population sample in rural Northeast China.
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Rac1 regulates skin tumors by regulation of keratin 17 through recruitment and interaction with CD11b+Gr1+ cells.
Oncotarget
PUBLISHED: 06-26-2014
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Rac1 is a member of the Rho family of small GTPases that control cells proliferation, differentiation, migration, and inflammation. Rac1 is crucial in tumorigenesis and development. Keratin17 and CD11b+Gr1+ cells are considered to regulate skin inflmmation. Here we discuss the regulation of Rac1 on skin tumor formation and its relationship. In samples from human skin squamous cell carcinoma (SCC), Rac1 activity was higher in cancer tissues than in normal skin and activity correlated with keratin 17 overexpression. In a DMBA/TPA-induced mouse skin tumor model, inhibition of Rac1 activity and depletion of CD11b+Gr1+ cells resulted in significant tumor formation. TPA induced recruitment of CD11b+Gr1+ cells into dermis; however, Rac1 inhibitor abolished this recruitment. In vitro, Rac1 induced interferon (IFN) and interlukin (IL6) production in keratinocytes, repression of keratin 17 inhibited IFN and IL6 production induced by Rac1. Moreover, both inhibition of Rac1 activity and repression of keratin 17 restricted proliferation and induction of differentiation in keratinocytes. Coculture of CD11b+Gr1+ cells with keratinocytes activated Wnt pathway in keratinocytes, resulting in enhanced Rac1 activity, overexpression of keratin 17, and hyperproliferation of keratinocytes. Our results suggested that hyperactive Rac1 recruited and interacted with CD11b+Gr1+ cells, inducing keratin 17-regulated inflammation and promoting skin tumor formation.
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Effects of histone deacetylase inhibitor oxamflatin on in vitro porcine somatic cell nuclear transfer embryos.
Cell Reprogram
PUBLISHED: 06-24-2014
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Low cloning efficiency is considered to be caused by the incomplete or aberrant epigenetic reprogramming of differentiated donor cells in somatic cell nuclear transfer (SCNT) embryos. Oxamflatin, a novel class of histone deacetylase inhibitor (HDACi), has been found to improve the in vitro and full-term developmental potential of SCNT embryos. In the present study, we studied the effects of oxamflatin treatment on in vitro porcine SCNT embryos. Our results indicated that the rate of in vitro blastocyst formation of SCNT embryos treated with 1 ?M oxamflatin for 15 h postactivation was significantly higher than all other treatments. Treatment of oxamflatin decreased the relative histone deacetylase (HDAC) activity in cloned embryos and resulted in hyperacetylation levels of histone H3 at lysine 9 (AcH3K9) and histone H4 at lysine 5 (AcH4K5) at pronuclear, two-cell, and four-cell stages partly through downregulating HDAC1. The suppression of HDAC6 through oxamflatin increased the nonhistone acetylation level of ?-tubulin during the mitotic cell cycle of early SCNT embryos. In addition, we demonstrated that oxamflatin downregulated DNA methyltransferase 1 (DNMT1) expression and global DNA methylation level (5-methylcytosine) in two-cell-stage porcine SCNT embryos. The pluripotency-related gene POU5F1 was found to be upregulated in the oxamflatin-treated group with a decreased DNA methylation tendency in its promoter regions. Treatment of oxamflatin did not change the locus-specific DNA methylation levels of Sus scrofa heterochromatic satellite DNA sequences at the blastocyst stage. Meanwhile, our findings suggest that treatment with HDACi may contribute to maintaining the stable status of cytoskeleton-associated elements, such as acetylated ?-tubulin, which may be the crucial determinants of donor nuclear reprogramming in early SCNT embryos. In summary, oxamflatin treatment improves the developmental potential of porcine SCNT embryos in vitro.
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Synthesis, antibacterial activities, and theoretical studies of dicoumarols.
Org. Biomol. Chem.
PUBLISHED: 06-21-2014
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Four dicoumarols (DC, 2-PyDC, 3-PyDC and 4-PyDC) were synthesized and characterized via IR, (1)H NMR, HRMS, and single crystal X-ray crystallography. Two classical intramolecular O-H···O hydrogen bonds (HBs) stabilized their structures. The total HB energies in DC, 2-PyDC, 3-PyDC and 4-PyDC were calculated with the density functional theory (DFT) [B3LYP/6-31G*] method. The in vitro antibacterial activity of DC, 2-PyDC, 3-PyDC and 4-PyDC against Staphylococcus aureus (S. aureus ATCC 29213), methicillin-resistant S. aureus (MRSA XJ 75302), vancomycin-intermediate S. aureus (Mu50 ATCC 700699), and USA 300 (Los Angeles County clone, LAC) was evaluated by observing the minimum inhibitory concentration and time-kill curves. The results showed that among all the compounds, 2-PyDC exhibited the most potent antibacterial activity.
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Rapid and sensitive screening and selective quantification of antibiotics in human urine by two-dimensional ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.
Anal Bioanal Chem
PUBLISHED: 05-24-2014
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A rapid and sensitive method for the screening and selective quantification of antibiotics in urine by two-dimensional ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was developed. This method allowed the injection of 200 ?L urine extract. The 200-?L injection volume used in this method increased the absolute sensitivity for target antibiotics in solvent by an average 13.3 times, with a range from 8.4 to 28.5 times, compared with the 10-?L conventional injection volume. A 96-well solid phase extraction procedure was established to eliminate the contamination on the chromatographic column resulting from the large-volume injection and increase the throughput of sample preparation. Fourteen target antibiotics from six common categories (?-lactams, quinolones, tetracyclines, macrolides, sulfonamides, and chloramphenicols) were selected as model compounds, and a database containing an additional 74 antibiotics was compiled for posttarget screening. The limit of detection of the target antibiotics, defined as a signal-to-noise ratio of 3, ranged from 0.04 to 1.99 ng/mL. The mean interday recoveries ranged between 79.6 and 121.3 %, with a relative standard deviation from 2.9 to 18.3 % at three spiking levels of 20 ng/mL, 50 ng/mL, and 100 ng/mL. This method was successfully applied in 60 real urine samples from schoolchildren aged 8-11 years, and four target antibiotics (azithromycin, sulfadiazine, trimethoprim, and oxytetracycline) and two posttarget antibiotics (sulfadimidine and cefaclor) were found in the urine samples. This method can be used as a large-scale biomonitoring tool for exposure of the human population to antibiotics.
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Ultrasensitive electrochemiluminescent detection of cardiac troponin I based on a self-enhanced Ru(II) complex.
Talanta
PUBLISHED: 05-14-2014
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To promote the luminous efficiency of luminophore, traditional electrochemiluminescence (ECL) immunoassay usually adopts the adding of coreactant into testing solution. However, many adverse micro-environmental factors in the solution are a limiting factor in ECL analytical techniques and received extensive attention. In our work, a self-enhanced ECL luminophore was synthesized by combining the coreactant (l-cysteine) and the luminophor (tris (4,4'-dicarboxylicacid-2,2'-bipyridyl) ruthenium(II) dichloride (Ru(dcbpy)3(2+))) to form one Ru(II) complex and was applied to fabricate a reagentless immunosensor for the detection of cardiac troponin I (cTnI) for the first time. Herein gold nanorods (AuNRs), due to their high specific surface area and good electrocatalytic ability, were used as carriers for the immobilization of Ru(II) complex and cTnI antibody to obtain the Ab2 bioconjugates as signal labels. The application of the self-enhanced Ru(II) complex not only avoided the addition of any coreactant into testing solution for simplifying the operation, but also achieved the intramolecular reaction for improving the ECL signal due to shorter electron transfer path and less energy loss. In view of these advantages, the proposed immunosensor achieved a wide linear range from 0.25 pg/mL to 0.1 ng/mL with an impressive detection limit of 0.083 pg/mL for cTnI (S/N=3).
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The Expression of CAP1 After Traumatic Brain Injury and Its Role in Astrocyte Proliferation.
J. Mol. Neurosci.
PUBLISHED: 05-08-2014
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Adenylate cyclase-associated protein 1 (CAP1), a member of cyclase-associated proteins involved in the regulation of actin filaments, was recently reported to play a role in the pathology of sciatic nerves injury. However, the distribution and function of CAP1 in the central nervous system (CNS) remain unclear. To investigate whether CAP1 is involved in CNS injury and repair, we used an acute traumatic brain injury (TBI) model in adult rats. Western blot analysis and immunohistochemistry showed a significant upregulation of CAP1 in ipsilateral peritrauma cortex compared with the contralateral and sham-operated ones. Double immunofluorescence staining showed that CAP1 was co-expressed with glial fibrillary acidic protein (GFAP). In addition, we detected that Ki-67 had colocalization with GFAP and CAP1 after TBI. In vitro, during the process of lipopolysaccharide (LPS)-induced primary astrocyte proliferation, we observed enhanced expression of CAP1. Specially, CAP1-specific siRNA-transfected primary astrocytes show significantly decreased ability for proliferation. Together, all these data indicated that the change of CAP1 protein expression was associated with astrocyte proliferation after the trauma of the central nervous system (CNS).
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Epigenetic changes of mesenchymal stem cells in three-dimensional (3D) spheroids.
J. Cell. Mol. Med.
PUBLISHED: 04-28-2014
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Mesenchymal stem cells (MSCs) hold profound promise in tissue repair/regeneration. However, MSCs undergo remarkable spontaneous differentiation and aging during monolayer culture expansion. In this study, we found that 2-3 days of three-dimensional (3D) spheroid culture of human MSCs (hMSCs) that had been expanded in monolayer for six passages increased their clonogenicity and differentiation potency to neuronal cells. Moreover, in accordance with these changes, the expression levels of miRNA which were involved in stem cell potency were changed and levels of histone H3 acetylation in K9 in promoter regions of Oct4, Sox2 and Nanog were elevated. Our results indicate that spheroid culture increases their multi-potency and changes the epigenetic status of pluripotent genes in hMSCs.
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Cotton (Gossypium hirsutum) 14-3-3 proteins participate in regulation of fibre initiation and elongation by modulating brassinosteroid signalling.
Plant Biotechnol. J.
PUBLISHED: 04-25-2014
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Cotton (Gossypium hirsutum) fibre is an important natural raw material for textile industry in the world. Understanding the molecular mechanism of fibre development is important for the development of future cotton varieties with superior fibre quality. In this study, overexpression of Gh14-3-3L in cotton promoted fibre elongation, leading to an increase in mature fibre length. In contrast, suppression of expression of Gh14-3-3L, Gh14-3-3e and Gh14-3-3h in cotton slowed down fibre initiation and elongation. As a result, the mature fibres of the Gh14-3-3 RNAi transgenic plants were significantly shorter than those of wild type. This 'short fibre' phenotype of the 14-3-3 RNAi cotton could be partially rescued by application of 2,4-epibrassinolide (BL). Expression levels of the BR-related and fibre-related genes were altered in the Gh14-3-3 transgenic fibres. Furthermore, we identified Gh14-3-3 interacting proteins (including GhBZR1) in cotton. Site mutation assay revealed that Ser163 in GhBZR1 and Lys51/56/53 in Gh14-3-3L/e/h were required for Gh14-3-3-GhBZR1 interaction. Nuclear localization of GhBZR1 protein was induced by BR, and phosphorylation of GhBZR1 by GhBIN2 kinase was helpful for its binding to Gh14-3-3 proteins. Additionally, 14-3-3-regulated GhBZR1 protein may directly bind to GhXTH1 and GhEXP promoters to regulate gene expression for responding rapid fibre elongation. These results suggested that Gh14-3-3 proteins may be involved in regulating fibre initiation and elongation through their interacting with GhBZR1 to modulate BR signalling. Thus, our study provides the candidate intrinsic genes for improving fibre yield and quality by genetic manipulation.
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Engaging academia to advance the science and practice of environmental public health tracking.
Environ. Res.
PUBLISHED: 04-25-2014
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Public health agencies at the federal, state, and local level are responsible for implementing actions and policies that address health problems related to environmental hazards. These actions and policies can be informed by integrating or linking data on health, exposure, hazards, and population. The mission of the Centers for Disease Control and Prevention?s National Environmental Public Health Tracking Program (Tracking Program) is to provide information from a nationwide network of integrated health, environmental hazard, and exposure data that drives actions to improve the health of communities. The Tracking Program and federal, state, and local partners collect, integrate, analyze, and disseminate data and information to inform environmental public health actions. However, many challenges exist regarding the availability and quality of data, the application of appropriate methods and tools to link data, and the state of the science needed to link and analyze health and environmental data. The Tracking Program has collaborated with academia to address key challenges in these areas. The collaboration has improved our understanding of the uses and limitations of available data and methods, expanded the use of existing data and methods, and increased our knowledge about the connections between health and environment. Valuable working relationships have been forged in this process, and together we have identified opportunities and improvements for future collaborations to further advance the science and practice of environmental public health tracking.
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Position-specific modification with imidazolyl group on10-23 DNAzyme realized catalytic activity enhancement.
Bioorg. Med. Chem.
PUBLISHED: 04-23-2014
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Nucleoside analogues with imidazolyl and histidinyl groups were synthesized for site-specific modification on the catalytic core of 10-23 DNAzyme. The distinct position-dependent effect of imidazolyl group was observed. Positive effect at A9 position was always observed. The pH- and Mg(2+)-dependence of the imidazolyl-modified DNAzymes suggested that imidazolyl group in 10-23 DNAzyme probably plays a dual role, its hydrogen bonding ability and spacial occupation play the favorable influence on the catalytic conformation of the modified DNAzymes. This research demonstrated that the catalytic performance of DNAzymes could be enhanced by incorporation of additional functional groups. Chemical modification is a feasible approach toward more efficient DNAzymes for therapeutic and biotechnological applications.
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'Imperfect' conjugated polymer nanoparticles from MEH-PPV for bioimaging and Fe(III) sensing.
Luminescence
PUBLISHED: 04-14-2014
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A simple and effective method was reported for the preparation from MEH-PPV of conjugated polymer nanoparticles (Pdots) that are water-soluble and well dispersed. The as-prepared Pdots show bright orange fluorescence at a quantum yield up to 32.37%. The fluorescence intensity of Pdots can be quenched with good selectively by the successive addition of Fe(3+) . In addition, the as-obtained Pdots were applied to the imaging of HeLa cells, and exhibited low cytotoxicity and excellent biocompatibility. Copyright © 2014 John Wiley & Sons, Ltd.
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Comprehensive optimization of emergency evacuation route and departure time under traffic control.
ScientificWorldJournal
PUBLISHED: 04-13-2014
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With the frequent occurrence of major emergencies, emergency management gets high attention from all around the world. This paper investigates the comprehensive optimization of major emergency evacuation route and departure time, in which case the evacuation propagation mechanism is considered under traffic control. Given the practical assumptions, we first establish a comprehensive optimization model based on the simulation of evacuation route and departure time. Furthermore, we explore the reasonable description method of evacuation traffic flow propagation under traffic control, including the establishment of traffic flow propagation model and the design of the simulation module that can simulate the evacuation traffic flow. Finally, we propose a heuristic algorithm for the optimization of this comprehensive model. In case analysis, we take some areas in Beijing as the evaluation sources to verify the reliability of our model. A series of constructive suggestions for Beijing's emergency evacuation are proposed, which can be applied to the actual situation under traffic control.
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Reversion of antibiotic resistance by inhibiting mecA in clinical methicillin-resistant Staphylococci by antisense phosphorothioate oligonucleotide.
J. Antibiot.
PUBLISHED: 04-09-2014
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Methicillin-resistant Staphylococci (MRS), methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE) have become a challenging problem in nosocomial infections and are connected with high morbidity and mortality rates. This is due to the increasing incidence of resistance to virtually all ?-lactams and a wide variety of antimicrobials. The spread of MRS severely limits therapeutic options and generates the need for novel antibiotics that are able to combat MRS infections. One method of inhibiting bacterial growth is by blocking the expression of conserved bacterial genes and provides potential new avenues for generating a new generation of antimicrobials. The mecA gene is highly conserved among Staphylococcal species, and this makes it an ideal target for antisense inhibition. We had identified a target sequence (854-871?nt) within the mecA mRNA coding region that is particularly sensitive to antisense inhibition. The anti-mecA PS-ODN04 oligonucleotide was encapsulated into an anionic liposome. MRSA01 and MRSE01 clinical strains treated with this antisense sequence became susceptible to existing ?-lactam antibiotics, and their growth was inhibited by oxacillin in vitro and in vivo. PS-ODN04 reduced the bacterial titers in the blood of mice infected with MRSA01 and MRSE01 and significantly improved their survival rate. Our data offer a possible new strategy for treating MRS infections.The Journal of Antibiotics advance online publication, 1 October 2014; doi:10.1038/ja.2014.132.
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Carbon dots as a luminescence sensor for ultrasensitive detection of phosphate and their bioimaging properties.
Luminescence
PUBLISHED: 03-23-2014
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Highly blue fluorescence carbon dots were synthesized by one-step hydrothermal treatment of potatoes. The as-obtained C-dots have been applied to bioimaging of HeLa cells, which shows their excellent biocompatibility and low cytotoxicity. The results reveal that C-dots are promising for real cell imaging applications. In addition, the carbon dots can be utilized as a probe for sensing phosphate. Copyright © 2014 John Wiley & Sons, Ltd.
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Matrix metalloproteinase-1 (MMP-1) expression in rat spinal cord injury model.
Cell. Mol. Neurobiol.
PUBLISHED: 03-22-2014
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Matrix metalloproteinase-1 (MMP-1), a member of the matrix metalloproteinases family, plays an integral role in extracellular matrix degradation and has been reportedly involved in the regulation of the brain or spinal cord traumatic neurovascular remodeling. Although the critical involvement of MMP-1 in the metastasis of tumors has been extensively documented, the role of MMP-1 in the pathology of neurological diseases remains largely elusive. In the present study, we established an adult rat spinal cord injury (SCI) model and investigated a potential role of MMP-1 in the pathological process of SCI. Using Western blot analysis, we identified notable expression change of MMP-1 after SCI. Immunohistochemistry showed that MMP-1 was distributed widely in rat spinal cord. Double immunofluorescence staining revealed that MMP-1 immunoreactivity was predominantly increased in neurons and astrocytes following SCI. Moreover, after injury, colocalization of MMP-1/active caspase-3 in neurons (NeuN-positive), and colocalization of MMP-1/PCNA in astrocytes (GFAP-positive) were clearly observed. We also examined the protein expression of PCNA, active caspase-3, Bcl-2, and Bax and found that the expression of the proteins was closely correlated with that of MMP-1. Taken together, our findings indicate that MMP-1 might play an important role in the regulation of neuronal apoptosis and astrocyte proliferation after SCI.
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The effect of dexamethasone on lentiviral vector infection is associated with importin ?
Biomed Rep
PUBLISHED: 03-21-2014
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Importin ? (Im?) plays an important role during the shuttling of the HIV-1 preintegration complex (PIC) from the cytoplasm to the nucleus. Im? may bind to the glucocorticoid receptor (GR), which is localized to nucleus following hormone binding. However, it remains unclear whether the binding of dexamethasone (Dex) to GR affects the Im? redistribution and, thus, alters PIC import. In our study, 293T cells were transfected with the lentiviral vector (LV) carrying the luciferase (Luci) gene following Dex or RU486 pretreatment. The Luci activity (LucA) in the Dex or RU486 group was significantly higher compared to that in the control group (P?0.01). The effects of Dex and RU486 were inhibited by the Im? inhibitor Bimax1 (P?0.01), although the inhibitory effect of Bimax1 was alleviated by increasing the Dex dose. Furthermore, it was observed that the LucA in the 30-min Dex treatment group was lower compared to that in the 30-min Dex pretreatment group (P?0.01). These results suggested that Dex may improve PIC import via increasing the cytoplasmic Im? levels. Kunming mice were transfected in vivo with the LV, either 30 min or 15 h following an intraperitoneal injection of Dex. The LucA in the liver of the 30-min group mice was significantly lower compared to that of the 15-h group mice (P?0.01), suggesting that the effect of Dex on LV infection depends mainly on the suppression of immune and inflammatory responses in vivo. Taken together, our data indicated that the effect of Dex on LV infection may be associated with Im?, constituting a novel signaling pathway mediating the effects of Dex on HIV-1 infection.
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Air pollutant emissions from on-road vehicles in China, 1999-2011.
Sci. Total Environ.
PUBLISHED: 03-09-2014
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The on-road vehicular emission in China from 1999 to 2011 was estimated, based on the emission factors of vehicles with different emission standards calculated by the COPERT model. The CO, NMVOC, NOX, BC and OC emissions changed from 19.7 Tg, 4.4 Tg, 2.3 Tg, 47.1 Gg and 74.4 Gg in 1999 to 32.7 Tg, 4.1 Tg, 7.6 Tg, 177.6 Gg and 101.5 Gg in 2011, respectively. The general trend for CO, NOX and BC was increasing, while the tendency for NMVOC and OC was firstly increase before 2002 and then decrease from 2003. The spatial analysis results showed that high emissions occurred in developed provinces (Guangdong, Shandong, Hebei, Jiangsu and Henan). The correlation between vehicular emissions and GDP were further investigated and good linear correlation was found. The not-obvious change of the inter-annual (1999-2011) fitted straight line slope and the sustained increasing emissions for NOX and BC suggested that the challenge of mitigating vehicular NOX and BC emissions is severe in China. The contribution from different vehicle types was also analyzed. Passenger car (PC) and motorcycle (MC) was the main contributor to the CO and NMVOC emissions. However, the contribution ratio of MC was decreasing from 36.6% and 68.8% in 1999 to 15.7% and 25.7% in 2011. Heavy duty truck (HDT) was the dominant contributor to NOX, BC and OC, with proportions of 58.9%, 57.6% and 52.8% in 2011, respectively. In addition, the uncertainty of the estimated emissions was also assessed based on the Monte Carlo simulation.
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The role of visfatin on the regulation of inflammation and apoptosis in the spleen of LPS-treated rats.
Cell Tissue Res.
PUBLISHED: 02-26-2014
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The purpose of the present study is to determine if visfatin is involved in inflammation or apoptosis induced by LPS in rat. Forty Wistar rats were divided into four groups: saline group, LPS group, visfatin group and Visfatin?+?LPS co-stimulated group. Spleen samples from each group of rats were collected for study. The spleen structure was examined by histological imaging. Apoptosis was evaluated with TUNEL reaction. Caspase-3 was detected with immunohistochemistry and western blot. The apoptosis-related genes were detected by qPCR and inflammatory cytokines were tested by ELISA. Our main findings were as follows. (1) Macrophages were markedly increased in the visfatin group compared with the saline group. This finding was confirmed when spleen samples were examined with western blot using CD68 antibody. (2) Visfatin promoted the expression of CD68 and caspase-3 in rat spleen, whereas visfatin could inhibit the expression of CD68 and activated caspase-3 in spleen of LPS-induced acute inflammation. (3) Visfatin had a pro-apoptotic effect on normal rat spleen, whereas it exerted an anti-apoptotic effect during LPS-induced lymphocytes apoptosis in rat spleen. Moreover, the effect of visfatin on cell apoptosis was mediated by the mitochondrial pathway. (4) Visfatin could modulate both the anti-inflammatory cytokines and pro-inflammatory cytokines in rat spleen, such as IL-10, IL-4, IL-6, TNF-? and IL-1?. Taken together, we demonstrate that visfatin could participate in the inflammatory process in rat spleen by modulating the macrophages and inflammatory cytokines. Also, visfatin plays a dual role in the apoptosis in rat spleen, which is mediated by the mitochondrial pathway.
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Development of a liquid chromatography-tandem mass spectrometry method for determination of butoconazole nitrate in human plasma and its application to a pharmacokinetic study.
J. Huazhong Univ. Sci. Technol. Med. Sci.
PUBLISHED: 02-21-2014
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A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 ?L were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 ?m), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8?165.1 for butoconazole and m/z 453.4?230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.
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Sphingosine-1-phosphate suppresses cyclophosphamide induced follicle apoptosis in human fetal ovarian xenografts in nude mice.
Fertil. Steril.
PUBLISHED: 02-20-2014
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To investigate the antiapoptosis effect of sphingosine-1-phosphate (S1P) on human fetal ovarian tissue treated by cyclophosphamide (CTX).
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Enhancement of electrogenerated chemiluminescence of luminol by ascorbic acid at gold nanoparticle/graphene modified glassy carbon electrode.
Spectrochim Acta A Mol Biomol Spectrosc
PUBLISHED: 02-19-2014
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Gold nanoparticle/graphene (GNP/GR) nanocomposite was one-pot synthesized from water soluble graphene and HAuCl? by hydrothermal method and characterized by TEM, Raman spectroscopy, XRD, XPS, UV-vis spectroscopy, and electrochemical impedance spectroscopy (EIS). Electrogenerated chemiluminescence (ECL) of luminol was investigated at the GNP/GR modified glassy carbon electrode (GNP/GR/GCE) and the GNP modified glassy carbon electrode (GNP/GCE) in aqueous solution respectively. The results revealed that one strong anodic ECL peak could be observed at ?0.8 V at two modified electrodes compared with that at the bare electrode. The intensity of the anodic ECL at the GNP/GR/GCE is weaker than that at the GNP/GCE, which should be due to the synergic effect of the enhancing effect of gold nanoparticles and the inhibiting effect of graphene on anodic luminol ECL. One strong cathodic ECL peak located at ?-0.8 V could be observed at the GNP/GR/GCE but not at the GNP/GCE, which should be result from the adsorbed oxygen at the graphene film. In the presence of ascorbic acid, the anodic ECL at the GNP/GR/GCE was enhanced more than 8-times, which is more apparent than that at the GNP/GCE. Whereas, the cathodic ECL peak was seriously inhibited at the GNP/GR/GCE. The enhanced ECL intensity at the GNP/GR/GCE varied linearly with the logarithm of ascorbic acid concentration in the range of 1.0 × 10(-8) to 1.0 × 10(-6)mol L(-1) with a detection limit of 1.0 × 10(-9) mol L(-1). The possible ECL mechanism was also discussed.
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Structure characterization and hypoglycemic effects of dual modified resistant starch from indica rice starch.
Carbohydr Polym
PUBLISHED: 02-18-2014
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Hypoglycemic effects of indica rice resistant starch (IR-RS) were investigated. We prepared IR-RS using a method that combined physical modification and enzyme modification, and the RS content was 47.0%. Differential scanning calorimetry--thermal gravimetric analysis showed that IR-RS have higher enthalpy and less loss of mass than single modified RS, heat-moisture RS and native starch. Scanning electron microscopy revealed that IR-RS displayed more compact spatial structure. IR-RS products displayed a mixture of B-and V-type x-ray diffraction patterns and the cyrstallinity was 51.0%. IR-RS significantly affected body weight, blood glucose, organ indices and serum lipid levels. These results demonstrated that dual modification changed the structure of indica rice starch and affected its digestibility as well as the blood glucose levels of the diabetic mice who consumed it.
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Docosahexanoic acid modifies low-density lipoprotein receptor abundance in HepG2 cells via suppression of the LXR??Idol pathway.
Mol Med Rep
PUBLISHED: 02-16-2014
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As a daily supplement, omega?3 fatty acid is confirmed to be of benefit in hypertriglyceridemia. However, the effect of omega?3 fatty acids on the low?density lipoprotein cholesterol (LDL?C) metabolism remains a controversial issue. In this study, we focused on the regulatory effect of docosahexanoic acid (DHA), one type of omega?3 fatty acid, exerted on the LDL receptor (LDLR), a determinant regulator of the LDL?C metabolism, and explored the potential mechanism. We observed that DHA increased hepatic LDLR protein in the presence of 25?hydroxycholesterol in HepG2 cells but did not alter the mRNA level. Previous studies have identified inducible degrader of the LDLR (Idol) as a novel negative post?translational modulator of LDLR and a direct transcriptional target of liver X receptor ? (LXR?). Since DHA had no effect on the transcriptional level of LDLR, we speculated that the post?transcriptional pathway LXR??Idol participated in this regulation. The results reveal that DHA downregulated the expression of LXR? and Idol in coordination with the upregulation of LDLR expression. Multiple mechanisms are involved in the regulation of LDLR by DHA, and the suppression of the LXR??Idol pathway is one of these mechanisms.
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Prediction of clinical pregnancy in vitrified-warmed single blastocyst transfer cycles by pre-freeze morphology.
Iran J Reprod Med
PUBLISHED: 02-15-2014
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Background: The selection of blastocyst warmed for transfer is based on pre-freeze morphology in vitrified-warmed single blastocyst transfer cycles. But, it is controversial which parameter of blastocyst morphology most closely related to the clinical outcomes. Objective: To estimate the effect of blastocoele expansion, trophectoderm (TE) morphology grade, and inner cell mass (ICM) morphology grade on clinical pregnancy in vitrified-warmed single blastocyst transfers. Materials and Methods: There were 172 vitrified-warmed single blastocyst transfer cycles during the year 2012 included in this analysis. Comparison of clinical results between pregnancy and no pregnancy group based on patient and blastocyst morphology characteristics was done. Then stepwise logistic regression analysis was used to select the best morphological predictor for clinical pregnancy. Last, comparison of patient characteristics and clinical outcomes separated by the best independent morphological predictor was done. Results: Comparison of clinical results between pregnancy and no pregnancy group and logistic regression showed the clinical pregnancy rate was affected by ICM. Comparison of patient characteristics separated by ICM grade, ICM grade A cycles got higher clinical pregnancy rate than ICM grade B cycles (54.3% vs. 35.0% respectively, p=0.037). Conclusion: Blastocyst with good ICM morphology could increase clinical pregnancy rate in vitrified-warmed single blastocyst transfer cycles.
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Downregulation of c-Met expression does not enhance the sensitivity of gastric cancer cell line MKN-45 to gefitinib.
Mol Med Rep
PUBLISHED: 01-24-2014
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The aim of the present study was to investigate the effect of downregulation of the c?Met gene on signal transduction and apoptosis in gastric cancer MKN?45 cells; furthermore, the study aimed to determine whether altered c?Met gene expression affected MKN?45 sensitivity to gefitinib. Three c?Met?specific small interfering RNAs (siRNAs) were synthesized and transfected into MKN?45 cells. Messenger RNA (mRNA) and protein levels of c?Met and its downstream signaling molecules [phosphoinositide 3?kinase (PI3K) and AKT] were examined using reverse transcription polymerase chain reaction and western blot analysis 48 h following transfection. Cell apoptosis was evaluated using Annexin?V/propidium iodide double staining and fluorescence?activated cell sorting analysis. An MTT assay was performed in order to measure the 50% inhibitory concentration (IC50) of gefitinib on MKN?45 cells. The results of the present study demonstrated that 48 h post?transfection with c?Met siRNA, MKN?45 cells showed significantly downregulated expression of c?Met mRNA and protein as well as an increased rate of apoptosis (P<0.05). In addition, following c?Met siRNA transfection mRNA and protein levels of PI3K and AKT were not significantly altered in MKN?45 cells (P>0.05); however, a marked decrease in the expression levels of phosphorylated (p)?PI3K and p?AKT was observed (P<0.05). Furthermore, the IC50 of gefitinib in MKN?45 cells was not significantly decreased. In conclusion, knockdown of the c?Met gene promoted gastric cancer cell apoptosis and inhibited downstream p?PI3K and p?AKT; however, the sensitivity of MKN?45 cells to gefitinib was not increased.
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Evaluation of humic substances during co-composting of food waste, sawdust and Chinese medicinal herbal residues.
Bioresour. Technol.
PUBLISHED: 01-24-2014
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Humification during co-composting of food waste, sawdust and Chinese medicinal herbal residues (CMHRs) was investigated to reveal its correlation with compost maturity. Food waste, sawdust and CMHRs were mixed at 5:5:1 and 1:1:1 (dry weight basis) while food waste:sawdust at 1:1 (dry wt. basis) served as control. Lime at 2.25% was added to all the treatments to alleviate low pH, and composted for 56 days. Humic acid/fulvic acid (HA/FA) ratio increased to 0.5, 2.0 and 3.6 in the control and treatment at 5:5:1, and 1:1:1 mixing ratio, respectively at the end of composting. The decrease in aliphatic organics in HA demonstrated the degradation of the readily available organics, while an increase in aromatic functional groups indicated the maturity of compost. Disappearance of hemicellulose and weak intensity of lignin in the CMHRs treatments indicated that the lignin provided the nucleus for HA formation; and the CMHRs accelerated the compost maturity.
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RASSF10 is epigenetically inactivated and induces apoptosis in lung cancer cell lines.
Biomed. Pharmacother.
PUBLISHED: 01-18-2014
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Ras-association domain family 10 (RASSF10), the latest member of the RASSF family with Ras effector function, has been frequently inactivated by aberrant promoter hypermethylation in several human cancers. However, its role in lung cancer has remained unclear. In this study, we investigated the methylation status of RASSF10 by combined bisulfate restriction analysis (COBRA) and examined its preliminary function in lung cancer cell lines. RASSF10 was methylated in four out of six lung cancer cell lines, including NCI-H157, NCI-460, SPCA-1 and NCI-H446. Treatment with a DNA methylation inhibitor, 5-aza-2'-deoxycytiding (5-aza-DC), restored RASSF10 mRNA expression and the restoration of RASSF10 increased cell apoptosis in a dose dependent manner, whereas knockdown of RASSF10 improved cell proliferation ability and inhibited cell apoptosis rate significantly. Immunofluorescence revealed that RASSF10 protein was located in the cell membrane. Taken together, our data for the first time demonstrates the frequent epigenetic inactivation of RASSF10 in lung cancer cell lines. RASSF10 induces cell apoptosis and might function as a tumor suppressor gene in lung cancer.
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The benzimidazole based drugs show good activity against T. gondii but poor activity against its proposed enoyl reductase enzyme target.
Bioorg. Med. Chem. Lett.
PUBLISHED: 01-09-2014
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The enoyl acyl-carrier protein reductase (ENR) enzyme of the apicomplexan parasite family has been intensely studied for antiparasitic drug design for over a decade, with the most potent inhibitors targeting the NAD(+) bound form of the enzyme. However, the higher affinity for the NADH co-factor over NAD(+) and its availability in the natural environment makes the NADH complex form of ENR an attractive target. Herein, we have examined a benzimidazole family of inhibitors which target the NADH form of Francisella ENR, but despite good efficacy against Toxoplasma gondii, the IC50 for T. gondii ENR is poor, with no inhibitory activity at 1 ?M. Moreover similar benzimidazole scaffolds are potent against fungi which lack the ENR enzyme and as such we believe that there may be significant off target effects for this family of inhibitors.
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Symmetrical acrokeratoderma: A peculiar entity in China? Clinicopathologic and immunopathologic study of 34 new cases.
J. Am. Acad. Dermatol.
PUBLISHED: 01-07-2014
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Symmetrical acrokeratoderma seems to be a new disorder in China, and 138 cases have been reported in the Chinese literature.
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Optimization of culture condition of human bone marrow stromal cells in terms of purification, proliferation, and pluripotency.
In Vitro Cell. Dev. Biol. Anim.
PUBLISHED: 01-05-2014
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Human bone marrow stromal cells (hBMSCs) possess multilineage differentiation potential and play an important role in modern tissue engineering. However, the development of culture media to maintain hBMSCs in an undifferentiated, self-renewing state during their robust proliferation remains a challenge. We developed and tested modified growth medium [medium 1: epidermal growth factor (EGF), platelet-derived growth factor (PDGF), low glucose, 2% fetal calf serum (FCS)] on hBMSCs by comparing primary cell isolation, multipassage expansion, culture morphology, proliferation, and cellular phenotype, and performing an expression analysis of intrinsic-regulated genes to other two media. Cell morphology, proliferation, and phenotype varied among the media, while cells cultured in medium 1 displayed small, spindle-shaped morphology with the highest rate of growth capacities and the expected phenotype. RT-PCR analysis showed that medium 1 displayed the lowest expression levels of osteogenic genes, chondrogenic genes (osteonectin, runt-related transcription factor 2, cartilage oligo matrix protein, and SOX9), and adipogenic genes (lipoprotein lipase). The expression of another adipogenic gene, peroxisome proliferator-activator receptor-?2, was higher in medium 1 but did not reach significance. In addition, hBMSCs expanded in medium 1 showed the highest expression ratio of self-renewing-related genes Krüppel-like factor 2 (KLF2) and KLF5. In conclusion, medium 1 allows for better expansion and pluripotency maintenance of hBMSCs and serves as a preferred alternative to traditional serum-containing media for research applications and future clinical use.
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Expression of hypoxia-inducible factor-1?, endothelin-1 and adrenomedullin in newborn rats with hypoxia-induced pulmonary hypertension.
Exp Ther Med
PUBLISHED: 01-04-2014
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Hypoxia-inducible factor (HIF)-1? is associated with hypoxia-induced pulmonary hypertension (HPH) in adults. In the present study, the expression levels of HIF-1?, endothelin (ET)-1 and adrenomedullin (ADM) were analyzed during HPH in neonates. In total, 96 newborn rats were subjected to hypoxia or normoxia for 3, 5, 7, 10, 14 or 21 days (n=8 per subgroup). HIF-1?, ET-1 and ADM expression levels were measured by quantitative polymerase chain reaction. In addition, the intima-media thickness/external diameter ratio (MT%) and medial wall cross-sectional area/vessel total cross-sectional area ratio (MA%) were calculated to evaluate pulmonary vascular remodeling. The mean pulmonary arterial pressure (mPAP) increased with exposure to hypoxia. Furthermore, the expression levels of HIF-1?, ET-1 and ADM in the lungs were shown to increase after three and five days of hypoxia, while the MT% and MA% increased after seven days of hypoxia, as compared with the controls (P<0.05). Therefore, the expression of HIF-1?, ET-1 and ADM is upregulated in the lungs of newborn rats during early HPH. At later stages, the mPAP increases, vascular remodeling occurs and HIF-1?, ET-1 and ADM expression levels restore to normal levels.
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Intravascular large B-cell lymphoma confirmed by lung biopsy.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
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Intravascular lymphoma is a very rare form of large B-cell non-Hodgkin's lymphoma which is characterized by selective growth of lymphoma cells within the lumina of small blood vessels. We report a 64-year-old woman visited hospital because of persistent cough, intermittent high fever as high as 38.7°C and occasional shortness of breath. Her chest CT showed left upper lobe pneumonia and tuberculosis skin test (PPD test) was positive. She was suspected with tuberculosis and treated with anti-tuberculosis drugs. However, her symptoms and general condition deteriorated, and she visited our hospital. She had no abnormal findings on physical examination, but had abnormal laboratory findings, including decreased hemoglobin, elevated LDH and C-reactive protein. Arterial blood gas analysis showed moderate hypoxaemia. A chest radiograph showed pneumonia in whole lung and CT showed diffused ground glass opacities in both lung fields. Lung biopsy confirmed a diagnosis of intravascular large B-cell lymphoma. Primary pulmonary manifestation is very rare. The diagnosis is based on the histopathology and immunohistochemistry.
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An estimating function approach to linkage heterogeneity.
J. Genet.
PUBLISHED: 12-28-2013
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Testing linkage heterogeneity between two loci is an important issue in genetics. Currently, there are four methods (K-test, A-test, B-test and D-test) for testing linkage heterogeneity in linkage analysis, which are based on the likelihood-ratio test. Among them, the commonly used methods are the K-test and A-test. In this paper, we present a novel test method which is different from the above four tests, called G-test. The new test statistic is based on estimating function, possessing a theoretic asymptotic distribution, and therefore demonstrates its own advantages. The proposed test is applied to analyse a real pedigree dataset. Our simulation results also indicate that the G-test performs well in terms of power of testing linkage heterogeneity and outperforms the current methods to some degree.
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Spiroindolone that inhibits PfATPase4 is a potent, cidal Inhibitor of Toxoplasma gondii tachyzoites in vitro and in vivo (132 characters, sp).
Antimicrob. Agents Chemother.
PUBLISHED: 12-23-2013
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Herein, we show that spiroindolone, an effective treatment of Plasmodia, is also active against T. gondii tachyzoites. In vitro, spiroindolone NITD 609 is cidal for tachyzoites (IC50= 1?M) and not toxic to human cells at ?10?M. Two daily oral doses of 100mg/Kg reduced parasite burden by 90% (p=0.002), measured 3 days after last dose. This inhibition of T. gondii tachyzoites in vitro and in vivo indicates spiroindolone is a promising lead candidate for further medicine development.
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Discrimination of Potent Inhibitors of Toxoplasma gondii Enoyl-Acyl Carrier Protein Reductase by a Thermal Shift Assay.
Biochemistry
PUBLISHED: 12-13-2013
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Many microbial pathogens rely on a type II fatty acid synthesis (FASII) pathway that is distinct from the type I pathway found in humans. Enoyl-acyl carrier protein reductase (ENR) is an essential FASII pathway enzyme and the target of a number of antimicrobial drug discovery efforts. The biocide triclosan is established as a potent inhibitor of ENR and has been the starting point for medicinal chemistry studies. We evaluated a series of triclosan analogues for their ability to inhibit the growth of Toxoplasma gondii, a pervasive human pathogen, and its ENR enzyme (TgENR). Several compounds that inhibited TgENR at low nanomolar concentrations were identified but could not be further differentiated because of the limited dynamic range of the TgENR activity assay. Thus, we adapted a thermal shift assay (TSA) to directly measure the dissociation constant (Kd) of the most potent inhibitors identified in this study as well as inhibitors from previous studies. Furthermore, the TSA allowed us to determine the mode of action of these compounds in the presence of the reduced nicotinamide adenine dinucleotide (NADH) or nicotinamide adenine dinucleotide (NAD(+)) cofactor. We found that all of the inhibitors bind to a TgENR-NAD(+) complex but that they differed in their dependence on NAD(+) concentration. Ultimately, we were able to identify compounds that bind to the TgENR-NAD(+) complex in the low femtomolar range. This shows how TSA data combined with enzyme inhibition, parasite growth inhibition data, and ADMET predictions allow for better discrimination between potent ENR inhibitors for the future development of medicine.
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Asparagine-linked glycosylation of bone morphogenetic protein-2 is required for secretion and osteoblast differentiation.
Glycobiology
PUBLISHED: 12-12-2013
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Bone morphogenetic protein-2 (BMP-2), a glycosylated protein, has been demonstrated to play a key role in osteoblast differentiation. However, the function of its glycosylation is incompletely understood. In this study, we examined the role that N-linked glycan play in the secretion of BMP-2. Blocking the addition of N-linked glycans (NLGs) or inhibiting initial glycan processing prevented the secretion of BMP-2. To identify the specific glycosylation sites, we abolished potential sites of N-linked glycosylation (Asn-Xaa-Ser/Thr) in BMP-2 by mutating the Asn residues to Gln individually or in combination, expressed the BMP-2 mutants in CHO and HEK293T cells and determined their glycosylation state by using PNGase F and Endo H digestion. We found that human BMP-2 contains three N-linked glycan on N135, N200 and N338. Elimination of N-glycosylation by mutation of N135 (N135Q) abolished the BMP-2 secretion from CHO cells. Overexpression of the BMP-2 mutant N135Q elicited endoplasmic reticulum (ER) stress and retained within the ER in CHO cells, indicating that N-glycosylation is required for folding of human BMP-2. Furthermore, we demonstrated that glycosylation at N135 was necessary for BMP-2 induced osteoblast differentiation in MC3T3-E1 cells. Taken together, these data provide further evidence of the critical role that individual N-linked glycan may play an important role in determining BMP-2 folding, secretion and function.
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[High-pitch dual-source CT in prospectively electrocardiogram-triggered spiral double scanning mode for diagnosing coronary artery patency].
Nan Fang Yi Ke Da Xue Xue Bao
PUBLISHED: 11-26-2013
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To assess the image quality, diagnostic accuracy and effective radiation dose of prospectively ECG- triggered high-pitch spiral double scanning (Double Flash) mode of computed tomography coronary angiography (CTCA) using dual-source CT for the diagnosis of significant coronary stenoses.
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Effect of the Fukushima nuclear accident on the risk perception of residents near a nuclear power plant in China.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 11-18-2013
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We assessed the influence of the Fukushima nuclear accident (FNA) on the Chinese publics attitude and acceptance of nuclear power plants in China. Two surveys (before and after the FNA) were administered to separate subsamples of residents near the Tianwan nuclear power plant in Lianyungang, China. A structural equation model was constructed to describe the public acceptance of nuclear power and four risk perception factors: knowledge, perceived risk, benefit, and trust. Regression analysis was conducted to estimate the relationship between acceptance of nuclear power and the risk perception factors while controlling for demographic variables. Meanwhile, we assessed the median public acceptable frequencies for three levels of nuclear events. The FNA had a significant impact on risk perception of the Chinese public, especially on the factor of perceived risk, which increased from limited risk to great risk. Public acceptance of nuclear power decreased significantly after the FNA. The most sensitive groups include females, those not in public service, those with lower income, and those living close to the Tianwan nuclear power plant. Fifty percent of the survey respondents considered it acceptable to have a nuclear anomaly no more than once in 50 y. For nuclear incidents and serious incidents, the frequencies are once in 100 y and 150 y, respectively. The change in risk perception and acceptance may be attributed to the FNA. Decreased acceptance of nuclear power after the FNA among the Chinese public creates additional obstacles to further development of nuclear power in China and require effective communication strategies.
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Outer insulation failure in a multilumen ICD lead.
Acta Cardiol
PUBLISHED: 11-06-2013
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Insulation defects in the leads are a common complication in implantable cardioverter-defibrillators (ICDs). Although the new multilumen leads have significantly reduced the incidence of insulation defects, they have introduced the new and rare problem of outer-insulation abrasions. In this report, we present the case of a 53-year-old man with dilated cardiomyopathy who was found to have an outer-insulation abrasion on fluoroscopy 3 years after the implantation of an ICD. Several electrical storms experienced by the patient were considered to be responsible for the abrasions; results of routine evaluations were normal. A CRT-D device was finally implanted.
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Effects of tanshinone on hyperandrogenism and the quality of life in women with polycystic ovary syndrome: protocol of a double-blind, placebo-controlled, randomised trial.
BMJ Open
PUBLISHED: 10-29-2013
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Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders in reproductive-age women. Chinese herbal medicine has been used for the treatment of PCOS, but the evidence for its efficacy and safety is minimal. Tanshinones are a class of bioactive molecules isolated from Salvia miltiorrhiza, a commonly used herb in Traditional Chinese Medicine. This study aims to evaluate the efficacy of tanshinones on hyperandrogenism and quality of life in women with PCOS who do not attempt to conceive.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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