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Find video protocols related to scientific articles indexed in Pubmed.
The Master Activator of IncA/C Conjugative Plasmids Stimulates Genomic Islands and Multidrug Resistance Dissemination.
PLoS Genet.
PUBLISHED: 10-01-2014
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Dissemination of antibiotic resistance genes occurs mostly by conjugation, which mediates DNA transfer between cells in direct contact. Conjugative plasmids of the IncA/C incompatibility group have become a substantial threat due to their broad host-range, the extended spectrum of antimicrobial resistance they confer, their prevalence in enteric bacteria and their very efficient spread by conjugation. However, their biology remains largely unexplored. Using the IncA/C conjugative plasmid pVCR94?X as a prototype, we have investigated the regulatory circuitry that governs IncA/C plasmids dissemination and found that the transcriptional activator complex AcaCD is essential for the expression of plasmid transfer genes. Using chromatin immunoprecipitation coupled with exonuclease digestion (ChIP-exo) and RNA sequencing (RNA-seq) approaches, we have identified the sequences recognized by AcaCD and characterized the AcaCD regulon. Data mining using the DNA motif recognized by AcaCD revealed potential AcaCD-binding sites upstream of genes involved in the intracellular mobility functions (recombination directionality factor and mobilization genes) in two widespread classes of genomic islands (GIs) phylogenetically unrelated to IncA/C plasmids. The first class, SGI1, confers and propagates multidrug resistance in Salmonella enterica and Proteus mirabilis, whereas MGIVmi1 in Vibrio mimicus belongs to a previously uncharacterized class of GIs. We have demonstrated that through expression of AcaCD, IncA/C plasmids specifically trigger the excision and mobilization of the GIs at high frequencies. This study provides new evidence of the considerable impact of IncA/C plasmids on bacterial genome plasticity through their own mobility and the mobilization of genomic islands.
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[Bacterial SXT/R391 family from integrating conjugative elements--a review].
Wei Sheng Wu Xue Bao
PUBLISHED: 09-10-2014
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SXT/R391 family has the most abundant types and members in integrating conjugative elements (ICE). SXT/ R391 elements are comprised of conservative core genes and genes in variable regions. The functions of conservative core genes of SXT/R391 include integration and excision, self-transfer through conjugation, and regulation of its expression. The genes in the variable regions often encode for drug and heavy metal resistances, forming of biofilm, adjustment of bacterial motility, and toxin-antitoxin systems that prevent SXT/R391 deletion from hosts. Some genes in variable region of SXT/R391 also encode for restriction-modification system, helicase, and endonuclease. The activity of SXT/R391 is positively regulated by activator SetCD, and negatively regulated by repressor SetR. SXT/R391 cannot be easily deleted from the primary donors in the process of transfer. SXT/R391 prevent the acquirement of closely related and homogeneous elements but cannot prevent the acquirement of heterogenetic ICE, which leads to the generation of hybrid ICE under the action of recombination system encoded by SXT/R391 themselves. SXT/R391 have high transferable frequency and wide host range, and until now more than 40 different SXT/R391 elements have been discovered in various bacteria, especially in Vibrio species, which mainly distribute in coastal areas in Asia and Africa. It suggests that marine environments are likely the main reservoir for SXT/R391 and these elements probably spread from marine environmental strains to clinical strains, under increasing selective pressure. Due to the hazard caused by the prevalence and the transfer of SXT/R391, medical microbiologist and health departments should be fully alert to the spread of the elements.
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Increased PTOV1 expression is related to poor prognosis in epithelial ovarian cancer.
Tumour Biol.
PUBLISHED: 09-03-2014
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Altered expression of prostate tumor overexpressed-1 (PTOV1) is observed in various types of human cancers. However, the role of PTOV1 in epithelial ovarian cancer (EOC) remains unclear. PTOV1 messenger (m)RNA expression in EOC patients was evaluated by quantitative real-time PCR (qRT-PCR). PTOV1 protein expression was also analyzed in archived paraffin-embedded EOC tissues using immunohistochemistry (IHC), and its association with overall survival of patients was analyzed by statistical analysis. Results from qRT-PCR analysis show that the expression level of PTOV1 mRNA was significantly higher in tumor tissues of EOC, compared to that in adjacent noncancerous tissues (P?
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IL-27 controls sepsis-induced impairment of lung antibacterial host defence.
Thorax
PUBLISHED: 07-29-2014
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Interleukin 27 (IL-27) is an important cytokine regulating host immune responses. However, its role in sepsis-induced immunosuppression remains unclear.
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Immunohistochemical mapping of neuropeptide Y in the tree shrew brain.
J. Comp. Neurol.
PUBLISHED: 06-24-2014
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Day-active tree shrews are promising animals as research models for a variety of human disorders. Neuropeptide Y (NPY) modulates many behaviors in vertebrates. Here we examined the distribution of NPY in the brain of tree shrews (Tupaia belangeri chinensis) using immunohistochemical techniques. The differential distribution of NPY-immunoreactive (-ir) cells and fibers were observed in the rhinencephalon, telencephalon, diencephalon, mesencephalon, metencephalon, and myelencephalon of tree shrews. Most NPY-ir cells were multipolar or bipolar in shape with triangular, fusiform, and/or globular perikarya. The densest cluster of NPY-ir cells were found in the mitral cell layer of the main olfactory bulb (MOB), arcuate nucleus of the hypothalamus, and pretectal nucleus of the thalamus. The MOB presented a unique pattern of NPY immunoreactivity. Laminar distribution of NPY-ir cells was observed in the MOB, neocortex, and hippocampus. Compared to rats, the tree shrews exhibited a particularly robust and widespread distribution of NPY-ir cells in the MOB, bed nucleus of the stria terminalis, and amygdala as well as the ventral lateral geniculate nucleus and pretectal nucleus of the thalamus. By contrast, a low density of neurons were scattered in the striatum, neocortex, polymorph cell layer of the dentate gyrus, superior colliculus, inferior colliculus, and dorsal tegmental nucleus. These findings provide the first detailed mapping of NPY immunoreactivity in the tree shrew brain and demonstrate species differences in the distribution of this neuropeptide, providing an anatomical basis for the participation of the NPY system in the regulation of numerous physiological and behavioral processes. J. Comp. Neurol., 2014. © 2014 Wiley Periodicals, Inc.
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Effect of intravenous gadolinium-DTPA on diffusion-weighted imaging of brain tumors: a short temporal interval assessment.
J Magn Reson Imaging
PUBLISHED: 06-14-2014
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To determine the effect of intravenous administration of gadolinium (Gd) contrast medium (Gd-DTPA) on diffusion-weighted imaging (DWI) for the evaluation of normal brain parenchyma vs. brain tumor following a short temporal interval.
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Inhibition of N-myc downstream-regulated gene-2 is involved in an astrocyte-specific neuroprotection induced by sevoflurane preconditioning.
Anesthesiology
PUBLISHED: 05-29-2014
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Mechanism of sevoflurane preconditioning-induced cerebral ischemic tolerance is unclear. This study investigates the role of N-myc downstream-regulated gene-2 (NDRG2) in the neuroprotection of sevoflurane preconditioning in ischemic model both in vivo and in vitro.
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Sirt3 attenuates hydrogen peroxide-induced oxidative stress through the preservation of mitochondrial function in HT22 cells.
Int. J. Mol. Med.
PUBLISHED: 05-24-2014
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Sirtuins (Sirt) are a family of phylogenetically conserved nicotinamide adenine nucleotide (NAD(+))-dependent protein deacetylases, among which Sirt3 resides primarily in the mitochondria and serves as a stress responsive deacetylase, playing a role in protecting cells from damage under stress conditions. The present study aimed to investigate the role of Sirt3 in hydrogen peroxide (H(2)O(2))-induced oxidative neuronal injury in HT22 mouse hippocampal cells. Treatment with H(2)O(2) increased the expression of Sirt3 in a dose- and time-dependent manner, and the knockdown of Sirt3 using specific small interfering RNA (siRNA) exacerbated the H(2)O(2)-induced neuronal injury. The overexpression of Sirt3 induced by lentiviral transfection significantly reduced the generation of reactive oxygen species (ROS) and lipid peroxidation following injury, whereas the activities of endogenous antioxidant enzymes were not affected. Further experiments revealed that the H(2)O(2)-induced inhibition of mitochondrial complex activity and adenosine triphosphate (ATP) synthesis, the decrease in mitochondrial Ca(2+) buffering capacity and mitochondrial swelling were all partly reversed by Sirt3. Furthermore, the overexpression of Sirt3 attenuated the release of cytochrome c, the increase in the Bax/Bcl-2 ratio, as well as caspase-9/caspase-3 activity induced by H(2)O(2), and eventually inhibited apoptotic neuronal cell death. These results suggest that Sirt3 acts as a prosurvival factor, playing an essential role in protecting HT22 cells under H(2)O(2)-induced oxidative stress, possibly by inhibiting ROS accumulation and the activation of the mitochondrial apoptotic pathway.
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Sirt3 protects cortical neurons against oxidative stress via regulating mitochondrial Ca2+ and mitochondrial biogenesis.
Int J Mol Sci
PUBLISHED: 04-27-2014
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Oxidative stress is a well-established event in the pathology of several neurobiological diseases. Sirt3 is a nicotinamide adenine nucleotide (NAD+)-dependent protein deacetylase that regulates mitochondrial function and metabolism in response to caloric restriction and stress. This study aims to investigate the role of Sirt3 in H2O2 induced oxidative neuronal injury in primary cultured rat cortical neurons. We found that H2O2 treatment significantly increased the expression of Sirt3 in a time-dependent manner at both mRNA and protein levels. Knockdown of Sirt3 with a specific small interfering RNA (siRNA) exacerbated H2O2-induced neuronal injury, whereas overexpression of Sirt3 by lentivirus transfection inhibited H2O2-induced neuronal damage reduced the generation of reactive oxygen species (ROS), and increased the activities of endogenous antioxidant enzymes. In addition, the intra-mitochondrial Ca2+ overload, but not cytosolic Ca2+ increase after H2O2 treatment, was strongly attenuated after Sirt3 overexpression. Overexpression of Sirt3 also increased the content of mitochondrial DNA (mtDNA) and the expression of mitochondrial biogenesis related transcription factors. All these results suggest that Sirt3 acts as a prosurvival factor playing an essential role to protect cortical neurons under H2O2 induced oxidative stress, possibly through regulating mitochondrial Ca2+ homeostasis and mitochondrial biogenesis.
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Activation of mGluR5 attenuates NMDA-induced neurotoxicity through disruption of the NMDAR-PSD-95 complex and preservation of mitochondrial function in differentiated PC12 cells.
Int J Mol Sci
PUBLISHED: 04-17-2014
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Glutamate-mediated toxicity is implicated in various neuropathologic conditions, and activation of ionotropic and metabotropic glutamate receptors is considered to be the most important mechanism. It has been reported that pharmacological saturation of metabotropic glutamate receptors (mGluRs) can facilitate N-methyl-D-aspartate receptor (NMDAR) related signaling cascades, but the mechanism leading to mGluR-NMDAR interactions in excitotoxic neuronal injury has remained unidentified. In the present study, we investigated the role of mGluR5 in the regulation of N-methyl-D-aspartate (NMDA)-induced excitotoxicity in differentiated PC12 cells. We found that activation of mGluR5 with the specific agonist R,S-2-chloro-5-hydroxyphenylglycine (CHPG) increased cell viability and inhibited lactate dehydrogenase (LDH) release in a dose-dependent manner. CHPG also inhibited an increase in the Bax/Bcl-2 ratio, attenuated cleavage of caspase-9 and caspase-3, and reduced apoptotic cell death after NMDA treatment. The NMDA-induced mitochondrial dysfunction, as indicated by mitochondrial reactive oxygen species (ROS) generation, collapse of mitochondrial membrane potential (MMP), and cytochrome c release, was also partly prevented by CHPG treatment. Furthermore, CHPG blocked the NMDA-induced interaction of NMDAR with postsynaptic density protein-95 (PSD-95), but had no effects on intracellular calcium concentrations. All these results indicated that activation of mGluR5 protects differentiated PC12 cells from NMDA-induced neuronal excitotoxicity by disrupting NMDAR-PSD-95 interaction, which might be an ideal target for investigating therapeutic strategies in various neurological diseases where excitotoxicity may contribute to their pathology.
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Complexity generation by chemical synthesis: a five-step synthesis of (-)-chaetominine from L-tryptophan and its biosynthetic implications.
Org. Biomol. Chem.
PUBLISHED: 03-29-2014
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We demonstrated, for the first time, that on the basis of chemistry principles, the hexacyclic peptidyl alkaloid (?)-chaetominine (1) can be synthesized in a straightforward manner from L-Trp. The approach features the efficient generation of molecular complexity via a tandem C3/C14 syn-selective epoxidation (dr = 3:2)–annulative ring-opening reaction and a regioselective epimerization at C14. The successful production of (?)-chaetominine (1) from L-Trp could be helpful for revealing how the configuration of L-tryptophan becomes inverted in the biosynthetic pathway of (?)-chaetominine (1).
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Chronic unilateral locked facet joint with spinal cord injury in a 26-month-old child: A case report.
J Spinal Cord Med
PUBLISHED: 03-29-2014
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Objectives This study presents the successful posterior surgical reduction and fusion on a 26-month-old child with chronic unilateral locked facet joint and spinal cord injury (SCI). Methods A 26-month-old child with chronic unilateral locked facet joint and SCI treated by posterior surgical reduction and fusion. Plaster external fixation was applied and rehabilitation exercise was trained post-operatively. Results Chronic unilateral locked facet joint was reduced successfully and bone fusion of C4/5 was achieved 3 months after surgery. The function of both lower limbs was improved 1 year after surgery, aided with physical rehabilitation. Conclusion Unilateral locked facet joint in pediatric population is rare. Few clinical experiences were found in the literature. Non-surgical treatment has advantages of not being invasive and is preferred for acute patients; however, it may not be suitable for chronic unilateral locked facet joint with SCI, in which surgical intervention is needed.
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Risk analysis of sulfites used as food additives in China.
Biomed. Environ. Sci.
PUBLISHED: 03-15-2014
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This study was to analyze the risk of sulfites in food consumed by the Chinese people and assess the health protection capability of maximum-permitted level (MPL) of sulfites in GB 2760-2011. Sulfites as food additives are overused or abused in many food categories. When the MPL in GB 2760-2011 was used as sulfites content in food, the intake of sulfites in most surveyed populations was lower than the acceptable daily intake (ADI). Excess intake of sulfites was found in all the surveyed groups when a high percentile of sulfites in food was in taken. Moreover, children aged 1-6 years are at a high risk to intake excess sulfites. The primary cause for the excess intake of sulfites in Chinese people is the overuse and abuse of sulfites by the food industry. The current MPL of sulfites in GB 2760-2011 protects the health of most populations.
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Characterization of two tartary buckwheat R2R3-MYB transcription factors and their regulation of proanthocyanidin biosynthesis.
Physiol Plant
PUBLISHED: 02-24-2014
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Tartary buckwheat (Fagopyrum tataricum Gaertn.) contains high concentrations of flavonoids. The flavonoids are mainly represented by rutin, anthocyanins and proanthocyanins in tartary buckwheat. R2R3-type MYB transcription factors (TFs) play key roles in the transcriptional regulation of the flavonoid biosynthetic pathway. In this study, two TF genes, FtMYB1 and FtMYB2, were isolated from F. tataricum and characterized. The results of bioinformatic analysis indicated that the putative FtMYB1 and FtMYB2 proteins belonged to the R2R3-MYB family and displayed a high degree of similarity with TaMYB14 and AtMYB123/TT2. In vitro and in vivo evidence both showed the two proteins were located in the nucleus and exhibited transcriptional activation activities. During florescence, both FtMYB1 and FtMYB2 were more highly expressed in the flowers than any other organ. The overexpression of FtMYB1 and FtMYB2 significantly enhanced the accumulation of proanthocyanidins (PAs) and showed a strong effect on the target genes' expression in Nicotiana tabacum. The expression of dihydroflavonol-4-reductase (DFR) was upregulated to 5.6-fold higher than that of control, and the expression level was lower for flavonol synthase (FLS). To our knowledge, this is the first functional characterization of two MYB TFs from F. tataricum that control the PA pathway.
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IFN- alpha blocks IL-17 production by peripheral blood mononuclear cells in patients with chronic active hepatitis B Infection.
BMC Infect. Dis.
PUBLISHED: 01-30-2014
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IFN-? has been used to treat patients with chronic active hepatitis B (CAHB). Recent studies have implicated the IL-23/Th-17 pathway in the pathogenesis of CAHB. In this study, we investigated whether IFN-? could affect this pathway.
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Hemin inhibits NLRP3 inflammasome activation in sepsis-induced acute lung injury, involving heme oxygenase-1.
Int. Immunopharmacol.
PUBLISHED: 01-22-2014
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NLRP3 inflammasome activation contributes to acute lung injury (ALI), accelerating caspase-1 maturation, and resulting in IL-1? and IL-18 over-production. Heme oxygenase-1 (HO-1) plays a protective role in ALI. This study investigated the effect of hemin (a potent HO-1 inducer) on NLRP3 inflammasome in sepsis-induced ALI. The sepsis model of cecal ligation and puncture (CLP) was used in C57BL6 mice. In vivo induction and suppression of HO-1 were performed by pretreatment with hemin and zinc protoporphyrin IX (ZnPP, a HO-1 competitive inhibitor) respectively. CLP triggered significant pulmonary damage, neutrophil infiltration, increased levels of IL-1? and IL-18, and edema formation in the lung. Hemin pretreatment exerted inhibitory effect on lung injury and attenuated IL-1? and IL-18 secretion in serum and lung tissue. In lung tissues, hemin down-regulated mRNA and protein levels of NLRP3, ASC and caspase-1. Moreover, hemin reduced malondialdehyde and reactive oxygen species production, and inhibited NF-?B and NLRP3 inflammasome activity. Meanwhile, hemin significantly increased HO-1 mRNA and protein expression and HO-1 enzymatic activity. In contrast, no significant differences were observed between the CLP and ZnPP groups. Our study suggests that hemin-inhibited NLRP3 inflammasome activation involved HO-1, reducing IL-1? and IL-18 secretion and limiting the inflammatory response.
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The four-step total synthesis of (-)-chaetominine.
Chem. Commun. (Camb.)
PUBLISHED: 01-14-2014
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The total synthesis of the alkaloid (-)-chaetominine (1) has been achieved in four steps with an overall yield of 33.4%. Key features of our strategy include a one-pot cascade indole epoxidation - epoxide ring-opening cyclization - lactamization reaction sequence, and the use of a nitro group as a latent amino group for the one-pot construction of the quinazolinone ring. This constitutes a step economical, redox economical and protecting group-free total synthesis.
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Interspinous spacer versus traditional decompressive surgery for lumbar spinal stenosis: a systematic review and meta-analysis.
PLoS ONE
PUBLISHED: 01-01-2014
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Dynamic interspinous spacers, such as X-stop, Coflex, DIAM, and Aperius, are widely used for the treatment of lumbar spinal stenosis. However, controversy remains as to whether dynamic interspinous spacer use is superior to traditional decompressive surgery.
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[The mRNA expression of mitogen-activated protein kinase signal pathway related genes in the blood of arseniasis patients caused by burning coal].
Zhonghua Yu Fang Yi Xue Za Zhi
PUBLISHED: 12-20-2013
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To detect the mRNA expression of ERK1, ERK2, JNK1 and P38 gene in mitogen-activated protein kinase(MAPK) path way in the arseniasis patients caused by burning coal.
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Draft Genome Sequence of the Opportunistic Marine Pathogen Vibrio harveyi Strain E385.
Genome Announc
PUBLISHED: 12-17-2013
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Vibrio harveyi strain E385 was isolated from a diseased cage-cultured grouper in Daya Bay, China. Phylogenetic analysis based on the 16S rRNA gene sequence showed similarity with V. harveyi strain BAA-1116. We sequenced the pathogenic strain V. harveyi E385 and compared the genome with that of the nonpathogenic strain V. harveyi BAA-1116.
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Blockade of SOCE protects HT22 cells from hydrogen peroxide-induced apoptosis.
Biochem. Biophys. Res. Commun.
PUBLISHED: 10-09-2013
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Oxidative stress is an established event in the pathology of neurobiological diseases. Previous studies indicated that store-operated Ca(2+) entry (SOCE) has been involved in oxidative stress. The present study was carried out to investigate the effects of SOCE inhibition on neuronal oxidative stress injury induced by hydrogen peroxide (H2O2) in HT22 cells, a murine hippocampal neuronal model. H2O2 insult induced significant intracellular Ca(2+) overload, mitochondrial dysfunction and cell viability decrease. Inhibition of SOCE by pharmacological inhibitor and STIM1 RNAi significantly alleviated intracellular Ca(2+) overload, restored the mitochondrial membrane potential (MMP), decreased cytochrome C release and eventually inhibited H2O2-induced cell apoptosis. These findings suggest that SOCE inhibition exhibited neuroprotection against oxidative stress induced by H2O2 and SOCE might be a useful therapeutic target in neurobiological disorders.
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[Molecular cloning of squalene synthase gene form Paris polyphylla and its expression in Escherichia coli].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 10-02-2013
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To clone the cDNA sequence of squalene synthase gene from Paris polyphylla, and characterize the biological features of the obtained SQS.
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Cloning of sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) gene from white shrimp, Litopenaeus vannamei and its expression level analysis under salinity stress.
Mol. Biol. Rep.
PUBLISHED: 09-14-2013
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Sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) is an intracellular membrane bound enzyme that utilizes the free energy of ATP to transport Ca(2+) against a concentration gradient. In the present study, a new SERCA gene (LvSERCA) from white shrimp (Litopenaeus vannamei) was cloned using suppression subtractive hybridization and rapid amplification of cDNA ends. The full-length cDNA of LvSERCA contained an open reading frame of 3,009 bp coding for 1,002 amino acids with a calculated molecular weight of approximately 109.8 kDa. The identity analysis of the amino acid sequence of LvSERCA showed that it is highly conserved with 10 transmembrane ?-helices, one P-domain, one A-domain and one N-domain. The phylogenetic analysis revealed that LvSERCA is similar to other Arthropoda SERCA proteins. The mRNA levels of LvSERCA under salinity stress (3 and 40 g L(-1)) were analyzed by reverse transcription PCR and quantitative real-time PCR. The results showed that LvSERCA was expressed in all tissues detected. LvSERCA mRNA levels were significantly higher under hyper-salinity than hypo-salinity. These results highlight that Ga(2+)-ATPase plays an essential role in adjustment salinity stress, which may be useful for selective breeding of L. vannamei.
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Seroprevalence of pertussis in China: Need to improve vaccination strategies.
Hum Vaccin Immunother
PUBLISHED: 09-09-2013
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Pertussis remains an important cause of infant death worldwide and is an ongoing public health concern even in countries with high vaccination coverage. A cross-sectional seroepidemiological study was undertaken to estimate true incidence rates and gain further insight into the epidemiology and burden of pertussis in China. During 2011, a total of 1080 blood samples were obtained from healthy individuals between 0 and 86 y of age in Zhengzhou, Central China. Serum IgG antibodies against pertussis toxin (PT) and filamentous hemagglutinin (FHA) were measured quantitatively using ELISA. The results showed that the geometric mean titers of PT and FHA IgG were 6.48 IU/mL (95% CI: 5.70-7.41 IU/mL) and 11.39 IU/mL (95% CI: 10.22-12.87 IU/mL) among subjects less than 4 y of age, indicating that pertussis antibody levels were low despite high vaccination coverage. Of the 850 subjects ?4 y of age, 56 (6.6%) had anti-PT IgG titers above 30 IU/mL, and 11 (1.3%) had antibodies titers above 80 IU/mL. The estimated age-specific incidence of infection with B. pertussis revealed a peak incidence in the 31 to 40 y age group, followed by the 41 to 60 y age group. Taken together, these results indicate that pertussis is common in Chinese subjects in Zhengzhou, especially in adults, suggesting that the disease burden is underestimated in China. Therefore, our study stresses the importance of strengthening the diagnostic capacity and improving surveillance system for delineating current epidemiological profiles of pertussis. Most importantly, it may be advisable to re-evaluate the current Chinese pertussis immunization schedule and implement to booster doses for older children, adolescents and adults.
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Aptamer biosensor for sensitive detection of toxin A of Clostridium difficile using gold nanoparticles synthesized by Bacillus stearothermophilus.
Biosens Bioelectron
PUBLISHED: 08-15-2013
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A sensitive electrochemical biosensor was developed to detect toxin A (TOA) of Clostridium difficile based on an aptamer selected by the systematic evolution of ligands using exponential enrichment and gold nanoparticles (GNPS) synthesized by Bacillus stearothermophilus. The thiolated single-stranded DNA used as the capture probe (CP) was first self-assembled on a Nafion-thionine-GNPS-modified screen-printed electrode (SPE) through an Au-thiol interaction. The horseradish peroxidase (HRP)-labeled aptamer probe (AP) was then hybridized to the complementary oligonucleotide of CP to form an aptamer-DNA duplex. In the absence of TOA, the aptamer-DNA duplex modified the electrode surface with HRP, so that an amperometric response was induced based on the electrocatalytic properties of thionine. This was mediated by the electrons that were generated in the enzymatic reaction of hydrogen peroxide under HRP catalysis. After the specific recognition of TOA, an aptamer-TOA complex was produced rather than the aptamer-DNA duplex, forcing the HRP-labeled AP to dissociate from the electrode surface, which reduced the catalytic capacity of HRP and reduced the response current. The reduction in the response current correlated linearly with the concentration of TOA in the range of 0-200ng/mL. The detection limit was shown to be 1nM for TOA. This biosensor was applied to the analysis of TOA and showed good selectivity, reproducibility, stability, and accuracy.
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Bioremediation and fodder potentials of two Sargassum spp. in coastal waters of Shenzhen, South China.
Mar. Pollut. Bull.
PUBLISHED: 07-27-2013
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In this study, the bioremediation potentials of two seaweeds (Sargassum hemiphyllum and S. henslowianum) against pollution in a coastal mariculture area of Shenzhen, South China, were investigated by comparing the growth, nutrient bioaccumulation capacity of plants from the seaweed bed (control site) with plants from the fish farm. Results indicated that both species are potential candidates for bioremediation in the fish farm areas in terms of their high growth rates and high bioaccumulation capacities on inorganic nutrients. Both Sargassum spp. contain high levels of crude protein (11.7-14.0%) and crude fat (2.2-2.7%), suggesting high nutritional values. The S. hemiphyllum may serve as a good aquaculture fodder with high nutritional compositions and low heavy metal contents. However, heavy metals (Cr, Pb and Cd) of S. henslowianum exceed the maximum allowable concentrations as aquatic feed, which restricts its fodder application. In general, the results of this study may contribute to the marine pollution bioremediation in the coastal areas of South China, especially in mariculture zones.
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High glucose induces activation of the local renin?angiotensin system in glomerular endothelial cells.
Mol Med Rep
PUBLISHED: 05-24-2013
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Activation of the intrarenal renin?angiotensin system (RAS), which has been identified in podocytes and mesangial cells, is a novel mechanism in the progression of diabetic kidney disease (DKD). The present study aimed to identify the local RAS in glomerular endothelial cells (GEnCs). Rat GEnCs were stimulated by culture medium containing 30 mmol/l glucose for 12, 24, 48 and 72 h. Angiotensin II (Ang II) concentrations in cell lysates and culture media were examined by ELISA and mRNA levels of angiotensinogen and renin in cell lysates were analyzed by quantitative polymerase chain reaction. Ang II type 1 receptor (AT1R), Ang II type 2 receptor (AT2R), renin and angiotensinogen levels in cell lysates were determined by western blot analysis. Localization of intracellular AT1R, AT2R, angiotensinogen and renin was identified by confocal immunofluorescence microscopy. Consequently, high glucose (HG) increased intracellular and extracellular Ang II levels. Captopril and chymostatin (inhibitor of chymase, an enzyme that converts Ang I to Ang II) were able to antagonize HG?induced Ang II generation. Moreover, HG increased angiotensinogen production in GEnCs and reduced renin mRNA expression without altering renin protein production. However, HG decreased AT1R levels and resulted in AT2R shifting from the nuclear to perinuclear region in GEnCs. In conclusion, HG activated the intracellular RAS in rat GEnCs and the underlying mechanism may involve angiotensin?converting enzyme (ACE) and non?ACE pathways. The effects of HG on GEnCs may also involve the substrate and receptors of Ang II.
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Homer1 knockdown protects dopamine neurons through regulating calcium homeostasis in an in vitro model of Parkinsons disease.
Cell. Signal.
PUBLISHED: 05-13-2013
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Homer1 protein is an important scaffold protein at postsynaptic density and has been demonstrated to play a central role in calcium signaling in the central nervous system. The aim of this study was to investigate the effects of Homer1 knockdown on MPP(+) induced neuronal injury in cultured dopamine (DA) neurons. We found that down-regulating Homer1 expression with specific small interfering RNA (siRNA) significantly suppressed LDH release, reduced Propidium iodide (PI) or Hoechst staining, increased the number of tyrosine hydroxylase (TH) positive cells and DA uptake, and attenuated apoptotic and necrotic cell death after MPP(+) injury. Homer1 knockdown decreased intracellular reactive oxygen species (ROS) generation through inhibition of intracellular calcium overload, but did not affect the endogenous antioxidant enzyme activities. Calcium imaging was used to examine the changes of intracellular Ca(2+) concentration ([Ca(2+)]cyt) and Ca(2+) in endoplasmic reticulum (ER) ([Ca(2+)]ER), and the results showed that Homer1 siRNA transfection attenuated ER Ca(2+) release up to 120min after MPP(+) injury. Furthermore, decrease of [Ca(2+)]cyt induced by Homer1 knockdown in MPP(+) treated neurons was further enhanced by NMDA receptor antagonists MK-801 and AP-5, but not canonical transient receptor potential (TRPC) channel antagonist SKF-96365. l-type calcium antagonist isradipine but not nimodipine further inhibited intracellular calcium overload after MPP(+) insult in Homer1 down-regulated neurons. These results suggest that Homer1 knockdown has protective effects against neuronal injury in in vitro PD model by reducing calcium overload mediated ROS generation, and this protection may be dependent at least in part on the regulatory effects on the function of calcium channels in both plasma membrane and ER.
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Salvianolic acid B induces apoptosis in human glioma U87 cells through p38-mediated ROS generation.
Cell. Mol. Neurobiol.
PUBLISHED: 05-12-2013
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Salvianolic acid B (SalB), the main water-soluble bioactive compounds isolated from the traditional Chinese medical herb Danshen, has been shown to exert anti-cancer effect in several cancer cell lines. The aim of our study was to investigate the potential anti-cancer effect of SalB in human glioma U87 cells. We found that treatment with SalB significantly decreased cell viability of U87 cells in a dose- and time-dependent manner. SalB also enhanced the intracellular ROS generation and induced apoptotic cell death in U87 cells. Western blot analysis suggested that SalB increased the phosphorylation of p38 MAPK and p53 in a dose-dependent manner. Moreover, blocking p38 activation by specific inhibitor SB203580 or p38 specific siRNA partly reversed the anti-proliferative and pro-apoptotic effects, and ROS production induced by SalB treatment. The anti-tumor activity of SalB in vivo was also demonstrated in U87 xenograft glioma model. All of these findings extended the anti-cancer effect of SalB in human glioma cell lines, and suggested that these inhibitory effects of SalB on U87 glioma cell growth might be associated with p38 activation mediated ROS generation. Thus, SalB might be concerned as an effective and safe natural anticancer agent for glioma prevention and treatment.
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The expression of hepatoma-derived growth factor in primary central nervous system lymphoma and its correlation with angiogenesis, proliferation and clinical outcome.
Med. Oncol.
PUBLISHED: 05-03-2013
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Hepatoma-derived growth factor (HDGF), a potential predictive and prognostic marker in several human cancers, is the firstly reported member of the HDGF family of proteins containing a well-conserved N-terminal amino acid sequence. HDGF is implicated in tumorigenesis by direct angiogenic activity, and its expression is correlated with aggressive biological ability of cancer cells including proliferation and angiogenesis. So, we propose that HDGF may be a valuable factor in progression and prognosis for primary central nervous system lymphoma (PCNSL) through its angiogenic and proliferative activity. So, HDGF, CD31 and Ki67 expression in the specimens of 60 patients suffering from PCNSL was investigated by immunohistochemistry in this study. Their correlations with clinicopathologic features and prognosis were evaluated to determine whether HDGF, CD31 and Ki67 expression levels correlate with the prognosis of the 60 patients suffering from PCNSL. We found that all PCNSL specimens showed HDGF, CD31 and Ki67 expression with different expression levels. Statistical analysis showed that HDGF had a positive correlation with CD31, but not with Ki67. Patients with higher HDGF and CD31 expression level had poorer overall survival rates than those with lower expression levels of HDGF and CD31, while Ki67 expression level did not correlate with overall survival. Multivariate analysis revealed that postoperative adjuvant chemotherapy and high expression of HDGF was independent prognostic indicator of patient survival.
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Downregulation of postsynaptic density-95-interacting regulator of spine morphogenesis reduces glutamate-induced excitotoxicity by differentially regulating glutamate receptors in rat cortical neurons.
FEBS J.
PUBLISHED: 04-30-2013
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Glutamate-induced excitotoxicity is involved in many neurological diseases. Preso, a novel postsynaptic scaffold protein, mediates excitatory synaptic transmission and various synaptic functions. In this study, we investigated the role of Preso in the regulation of glutamate-induced excitotoxicity in rat cortical neurons. Knockdown of Preso with small interfering RNA improved neuronal viability and attenuated the elevation of lactate dehydrogenase (LDH) release after glutamate treatment. Downregulation of Preso also inhibited an increase in the BAX/Bcl-2 ratio and cleavage of caspase-9 and caspase-3. Although the expression and distribution of metabotropic glutamate receptor (mGluR) 1/5, NR1, NR2A and NR2B were not changed by knockdown of Preso, downregulation of Preso protected neurons from glutamate-induced excitotoxicity by inhibiting mGluR and N-methyl-d-aspartate receptor function. However, downregulation of Preso neither affected the expression of GluR1 and GluR2 nor influenced the function of ?-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor after glutamate treatment. Furthermore, intracellular Ca(2+) was an important downstream effector of Preso in the regulation of excitotoxicity. These results suggest that expression of Preso promotes the induction of excitotoxicity by facilitating different glutamate receptor signaling pathways. Therefore, Preso might be a potential pharmacological target for preventing and treating neurological diseases.
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Pseudoalteromonas xishaensis sp. nov., isolated from Acanthaster planci in the Xisha islands.
Antonie Van Leeuwenhoek
PUBLISHED: 04-16-2013
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A Gram-negative, aerobic, motile by means of single polar flagellum, short rod-shaped marine bacterium, designated strain E418T, was isolated from the spines on the body surface of starfish Acanthaster planci in the Xisha islands, China. Cells of strain E418T were found to grow optimally at pH 7–8, at 25–37 °C, and in the presence of 2–5 % (w/v) NaCl. Phylogenetic analysis based on the comparison of 16S rRNA gene sequences revealed that strain E418T is a member of the genus Pseudoalteromonas. The closest relative to this strain was found to be P. ruthenica LMG 19699T, with a similarity level of 97.7 %. DNA relatedness between the novel isolate and this phylogenetically related species was 57.4 %. Strain E418T decomposed Tween 80, gelatin, and casein, but was unable to decompose starch and grow on DNase Agar. The cellular fatty acid profile consisted of significant amounts of C16:1?7c/C16:1?6c, C18:1?7c/C18:1?6c, C16:0, and C17:1?8c. The G+C content of DNA of this strain was determined to be 46.7 mol%. Phenotypic characteristics, phylogenetic analysis and DNA–DNA relatedness data suggest that strain E418T represents a novel species of the genus Pseudoalteromonas, for which the name Pseudoalteromonas xishaensis sp. nov. is proposed. The type strain of P. xishaensis is strain E418T (DSM 25588T = NBRC 108846T = CCTCC AB 2011177T).
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Bayesian analyses of time-interval data for environmental radiation monitoring.
Health Phys
PUBLISHED: 03-21-2013
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Time-interval (time difference between two consecutive pulses) analysis based on the principles of Bayesian inference was investigated for online radiation monitoring. Using experimental and simulated data, Bayesian analysis of time-interval data [Bayesian (ti)] was compared with Bayesian and a conventional frequentist analysis of counts in a fixed count time [Bayesian (cnt) and single interval test (SIT), respectively]. The performances of the three methods were compared in terms of average run length (ARL) and detection probability for several simulated detection scenarios. Experimental data were acquired with a DGF-4C system in list mode. Simulated data were obtained using Monte Carlo techniques to obtain a random sampling of the Poisson distribution. All statistical algorithms were developed using the R Project for statistical computing. Bayesian analysis of time-interval information provided a similar detection probability as Bayesian analysis of count information, but the authors were able to make a decision with fewer pulses at relatively higher radiation levels. In addition, for the cases with very short presence of the source (< count time), time-interval information is more sensitive to detect a change than count information since the source data is averaged by the background data over the entire count time. The relationships of the source time, change points, and modifications to the Bayesian approach for increasing detection probability are presented.
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Long-term effect of vasectomy on spermatogenesis in men: a morphometric study.
Asian J. Androl.
PUBLISHED: 02-25-2013
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Spermatogenic damage may occur after vasectomy, and the damage is pressure mediated, occurring when the occluded reproductive tract is unable to accommodate additional sperm produced by the testis. This study aimed to determine the long-term effect of vasectomy on spermatogenesis in humans and clarify how the balance between sperm production in the testis and sperm storage in or removal from the tract might be maintained. During inguinal hernia repair, an open biopsy was performed to obtain testicular tissue blocks from 51 Chinese men (aged ?50 years), of whom 25 (control group) had not undergone vasectomy and 26 (vasectomized group) had undergone bilateral vasectomy 22-42 years before. Methacrylate resin-embedded testicular sections were made, and morphometric studies were performed using light microscopy. In addition, sizes of the testis and epididymis were estimated with ultrasonography. The testicular tissue blocks obtained from one control and seven vasectomized men consisted almost completely of connective tissue. In the other 43 men, significant differences were not found between the two groups in the testicular or epididymal size, qualitative histology or quantitative parameters including the mean diameter or volume fraction of the seminiferous tubules. In conclusion, sperm production and sperm storage/removal reached a static equilibrium after vasectomy, likely due to spermatogenic degeneration or less sperm production as a result of aging or due to vasectomy-induced testicular (interstitial) fibrosis. Thus, complications that might occur in association with overproduction of sperm and distension of the tract would disappear or be relieved with time.
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Glucagon-like peptide-1 protects against cardiac microvascular injury in diabetes via a cAMP/PKA/Rho-dependent mechanism.
Diabetes
PUBLISHED: 01-30-2013
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Impaired cardiac microvascular function contributes to cardiovascular complications in diabetes. Glucagon-like peptide-1 (GLP-1) exhibits potential cardioprotective properties in addition to its glucose-lowering effect. This study was designed to evaluate the impact of GLP-1 on cardiac microvascular injury in diabetes and the underlying mechanism involved. Experimental diabetes was induced using streptozotocin in rats. Cohorts of diabetic rats received a 12-week treatment of vildagliptin (dipeptidyl peptidase-4 inhibitor) or exenatide (GLP-1 analog). Experimental diabetes attenuated cardiac function, glucose uptake, and microvascular barrier function, which were significantly improved by vildagliptin or exenatide treatment. Cardiac microvascular endothelial cells (CMECs) were isolated and cultured in normal or high glucose medium with or without GLP-1. GLP-1 decreased high-glucose-induced reactive oxygen species production and apoptotic index, as well as the levels of NADPH oxidase such as p47(phox) and gp91(phox). Furthermore, cAMP/PKA (cAMP-dependent protein kinase activity) was increased and Rho-expression was decreased in high-glucose-induced CMECs after GLP-1 treatment. In conclusion, GLP-1 could protect the cardiac microvessels against oxidative stress, apoptosis, and the resultant microvascular barrier dysfunction in diabetes, which may contribute to the improvement of cardiac function and cardiac glucose metabolism in diabetes. The protective effects of GLP-1 are dependent on downstream inhibition of Rho through a cAMP/PKA-mediated pathway.
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Distribution and pathogenic relationship of virulence associated genes among Vibrio alginolyticus from the mariculture systems.
Mol. Cell. Probes
PUBLISHED: 01-14-2013
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Vibrio alginolyticus has been confirmed as an important pathogen for aquatic animals. However, the pathogenic mechanism of V. alginolyticus is not completely understood. A total of 31 isolates of V. alginolyticus from sea water, fish and shrimp on the mariculture systems were fingerprinted by pulsed-field gel electrophoresis. The pathogenicity of these isolates was tested by challenge and the 21 genes associated with the virulence of Vibrio cholerae or Vibrio parahaemolyticus were examined in V. alginolyticus using PCR. The results showed that the 31 V. alginolyticus isolates belonged to 26 PFGE genotypes and the isolates from different source had different genotypes. Nine of the 31 isolates were confirmed as pathogenic strains by challenge. Moreover, 12 vibrio virulence genes were detected in this study. Of the detected genes, VCtoxR, VCtoxS, hlyA, VPtoxR and tlh were found in both pathogenic and non-pathogenic isolates. However, the other 7 virulence genes, ctxB, zot, tagA, stn, sto, tdh and trh, were only present in pathogenic isolates. Analysis of the relationship between virulence associated genes and pathogenicity of V. alginolyticus provides a possible explanation that the pathogenic mechanism of V. alginolyticus might be similar to that of V. parahaemolyticus instead of V. cholerae.
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Iduna protects HT22 cells from hydrogen peroxide-induced oxidative stress through interfering poly(ADP-ribose) polymerase-1-induced cell death (parthanatos).
Cell. Signal.
PUBLISHED: 01-08-2013
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Oxidative stress-induced cell death is common in many neurological diseases. However, the role of poly(ADP-ribose) polymerase-1-induced cell death (parthanatos) has not been fully elucidated. Here, we found that hydrogen peroxide (H2O2) could lead to PARP-1 activation and apoptosis-inducing factor nuclear translocation in a concentration dependent manner. Iduna, as a novel regulator of parthanatos, was also induced by H2O2. Down-regulation of Iduna by genetic ablation promoted H2O2-induced cell damage. Up-regulation of Iduna reduced the loss of mitochondrial potential and ATP and NAD+ production, but did not affect the mitochondrial dysfunction-induced cytochrome c release, increase of Bax/Bcl-2 ratio, and Caspase-9/Caspase-3 activity. In contrast, overexpression of Iduna inhibited activation of PARP-1 and nuclear translocation of AIF. Further study showed that PARP-1 specific inhibitor, DPQ, blocked the protective effect of Iduna against H2O2-induced oxidative stress. Moreover, in the presence of proteasome inhibitor (MG-132) or ubiquitin E1 inhibitor (PYR-41), protective effect of Iduna was significantly weaken. These results indicate that Iduna acts as a potential antioxidant by improving mitochondrial function and inhibiting oxidative stress-induced parthanatos, and these protective effects are dependent on the involvement of ubiquitin-proteasome system.
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Vibrio zhuhaiensis sp. nov., isolated from a Japanese prawn (Marsupenaeus japonicus).
Antonie Van Leeuwenhoek
PUBLISHED: 01-07-2013
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A Gram-negative, oxidase-positive, facultatively anaerobic bacterium, designated strain E20121, was isolated from the digestive tract of a Japanese prawn (Marsupenaeus japonicus) collected from the coastal sea water area of Zhuhai, Guangdong province, China. The new isolate was determined to be closely related to Vibrio ponticus DSM 16217(T), having 97.6 % 16S rRNA gene sequence similarity. Phylogenetic analysis based on recA, pyrH and rpoA also showed low levels of sequence similarities (72.6-96.6 %) with all species of the genus Vibrio. A multigene phylogenetic tree using concatenated sequences of the four genes (16S rRNA, rpoA, recA and pyrH) clearly showed that the new isolate is different from the currently known Vibrio species. DNA-DNA hybridization experiments revealed similarity values below 70 % with the closest related species V. ponticus DSM 16217(T). Several phenotypic traits enabled the differentiation of strain E20121 from the closest phylogenetic neighbours. The DNA G+C content of strain E20121 was determined to be 47.6 mol % and the major fatty acid components identified were C16:1?7c and/or C16:1?6c (39.8 %), C18:1?7c (13.6 %) and C16:0 (9.6 %). Based on genotypic, phenotypic, chemotaxonomic, phylogenetic and DNA-DNA hybridization analyses, strain E20121 is proposed to represent a novel species of the genus Vibrio for which the name Vibrio zhuhaiensis sp. nov. is proposed. The type strain is E20121(T)(=DSM 25602(T) = CCTCC AB 2011174(T)).
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Saponin 1 Induces Apoptosis and Suppresses NF-?B-Mediated Survival Signaling in Glioblastoma Multiforme (GBM).
PLoS ONE
PUBLISHED: 01-01-2013
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Saponin 1 is a triterpeniod saponin extracted from Anemone taipaiensis, a traditional Chinese medicine against rheumatism and phlebitis. It has also been shown to exhibit significant anti-tumor activity against human leukemia (HL-60 cells) and human hepatocellular carcinoma (Hep-G2 cells). Herein we investigated the effect of saponin 1 in human glioblastoma multiforme (GBM) U251MG and U87MG cells. Saponin 1 induced significant growth inhibition in both glioblastoma cell lines, with a 50% inhibitory concentration at 24 h of 7.4 µg/ml in U251MG cells and 8.6 µg/ml in U87MG cells, respectively. Nuclear fluorescent staining and electron microscopy showed that saponin 1 caused characteristic apoptotic morphological changes in the GBM cell lines. Saponin 1-induced apoptosis was also verified by DNA ladder electrophoresis and flow cytometry. Additionally, immunocytochemistry and western blotting analyses revealed a time-dependent decrease in the expression and nuclear location of NF-?B following saponin 1 treatment. Western blotting data indicated a significant decreased expression of inhibitors of apoptosis (IAP) family members,(e.g., survivin and XIAP) by saponin 1. Moreover, saponin 1 caused a decrease in the Bcl-2/Bax ratio and initiated apoptosis by activating caspase-9 and caspase-3 in the GBM cell lines. These findings indicate that saponin 1 inhibits cell growth of GBM cells at least partially by inducing apoptosis and inhibiting survival signaling mediated by NF-?B. In addition, in vivo study also demonstrated an obvious inhibition of saponin 1 treatment on the tumor growth of U251MG and U87MG cells-produced xenograft tumors in nude mice. Given the minimal toxicities of saponin 1 in non-neoplastic astrocytes, our results suggest that saponin 1 exhibits significant in vitro and in vivo anti-tumor efficacy and merits further investigation as a potential therapeutic agent for GBM.
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Results of intercostal nerve transfer to the musculocutaneous nerve in brachial plexus birth palsy.
J Pediatr Orthop
PUBLISHED: 11-22-2011
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Intercostal nerve (ICN) transfer has been one of the main extraplexal nerve transfers in treating brachial plexus root avulsion. This retrospective study evaluated results of ICN transfer for reconstruction of the musculocutaneous nerve (MCN) in brachial plexus birth palsy (BPBP).
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1-Methyl-3-trifluoro-methyl-5-[(3-chloro-phen-yl)sulfanyl]-1H-pyrazole-4-carbaldehyde O-(4-chloro-benzo-yl)oxime.
Acta Crystallogr Sect E Struct Rep Online
PUBLISHED: 11-04-2011
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In the title compound, C(19)H(12)Cl(2)F(3)N(3)O(2)S, the 3-chloro-phenyl and 4-chloro-phenyl rings form dihedral angles 89.5?(2) and 11.4?(2)°, respectively, with the pyrazole ring. In the crystal, mol-ecules related by translation along the a axis are linked into chains via C-H?N hydrogen bonds.
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[Analyze on HPLC fingerprint of four main metropolis of herba pogostemonis].
Zhong Yao Cai
PUBLISHED: 10-25-2011
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To establish a method for analyzing HPLC fingerprint of Herba pogostemonis and compare the variability of four main producing areas.
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5-(3-Chloro-phen-ylsulfan-yl)-1-methyl-3-trifluoro-methyl-1H-pyrazole-4-carbaldehyde O-[(2-chloro-1,3-thia-zol-5-yl)meth-yl]oxime.
Acta Crystallogr Sect E Struct Rep Online
PUBLISHED: 10-17-2011
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In the title compound, C(16)H(11)Cl(2)F(3)N(4)OS(2), the benzene ring and the thia-zole ring make dihedral angles of 83.2?(3) and 78.3?(3)°, respectively, with the pyrazole ring. The crystal packing shows S?N contacts of 3.309?(2)?Å.
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Vibrio zhanjiangensis sp. nov., isolated from sea water of shrimp farming pond.
Antonie Van Leeuwenhoek
PUBLISHED: 08-31-2011
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A Gram-negative, facultatively anaerobic, motile by means of single polar flagellum, rod-shaped marine bacterium, designated strain E414, was isolated from sea water collected from a farming pond rearing marine shrimp Litopenaeus vannamei in Zhanjiang, Guangdong province, PRC. The strain was able to grow in the presence of 0.5-6% (w/v) NaCl (optimally in 3-6% (w/v) NaCl), between pH 6 and 9 (optimally at pH 7-8), between 15 and 37°C (optimally at 25-30°C). Phylogenetic analysis based on 16S rRNA gene sequences locate strain E414 in the vicinity of the coralliilyticus clade within the genus Vibrio. DNA-DNA relatedness data and multigene phylogenetic analysis based on the concatenated sequences of four genes (16S rRNA, rpoA, recA and pyrH) clearly differentiated strain E414 from its closest phylogenetic neighbours. Analysis of phenotypic features, including enzyme activities and utilization and fermentation of various carbon sources, further revealed discrimination between strain E414 and phylogenetically related Vibrio species. The major fatty acid components are C(16:1)?6c and/or C(16:1)?7c (27.4%), C(18:1)?7c and/or C(18:1)?6c (19.3%) and C(16:0) (18.2%). The DNA G+C content of strain E414 was 38.7 mol%. Based on phenotypic, chemotaxonomic, phylogenetic and DNA-DNA relatedness values, it can be concluded that E414 should be placed in the genus Vibrio as representing a novel species, for which the name Vibrio zhanjiangensis sp. nov. is proposed, with the type strain E414 (=CCTCC AB 2011110(T) = NBRC 108723(T) = DSM 24901).
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[Diversity and bacteria community structure of activated carbon used in advanced drinking water treatment].
Huan Jing Ke Xue
PUBLISHED: 07-26-2011
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Two granular activated carbon (GAC) samples with 1.5 a and 5 a age were collected, Bacterial genome DNA was extracted for the 16S rDNA gene amplification, and then a bacterial 16S rDNA gene clone library was constructed. After the phylogenetic analysis of 16S rDNA sequences, bacterial diversity and community structure of two activated carbon biofilm sample were studied. The results showed the bacteria in GAC with 5 a age could be divided into 11 groups, which were as follows alpha-Proteobacteria (26.5%), beta-Proteobacteria (16.3%), delta-Proteobacteria (16.3%), Planctomycetes (12.2%), Gemmatimonadetes (6.1%), Acidobacteria (4.1%), Nitrospira (2.0%), gamma-Proteobacteria (2.0%), Bacteroidetes (2.0%), Actinobacteria (2.0%), Unclassified Bacteria (10.2%). The bacteria in GAC with 1.5 a age could be divided into 10 groups, which were as follows alpha-Proteobacteria (21.6%), Planctomycetes( 10.8%), Bacteroidetes (10.8%), beta-Proteobacteria (9.0%), Acidobacteria (9.0%), Nitrospira (7.2%), detla-Proteobacteria (7.2%), Unclassified Proteobacteria (5.4%), Gemmatimonadetes (3.6%), Unclassified Bacteria (14.4%). The results revealed a variety of bacterial divisions on the studied GAC biofilm. Proteobacteria had the highest share in the two total clones, and alpha- and beta-Proteobacteria were on a dominant position. A relatively high proportion of delta-Proteobacteria was observed in the biofilm of GAC with 5 a age, and Nitrospira was in a minor proportion. However, a totally converse condition appeared in GAC with 1.5 a age. Two pathogenic bacteria, Afipia and Chryseobacterium, were detected in analyzed GACs, which implies a potential microbial risk in water supply.
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Isolation, purification and structure identification of two phenolic glycosides from the roots of Incarvillea younghusbandii Sprague and their antioxidant activities.
Yao Xue Xue Bao
PUBLISHED: 07-14-2011
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Using a bioassay-guided fractionation technique, two compounds were isolated from the roots of Incarvillea younghusbandii Sprague through silica gel, reverse-phase C18 column chromatography and reverse-phase HPLC. Their structures were identified as acteoside (1) and isoacteoside (2) by ESI-MS, GC-MS, 1D- and 2D-NMR. 1 and 2 showed *OH scavenging capacity similar with benzoic acid, higher O2*- (or *OH) scavenging capacity than ascorbic acid, far higher hepatic LPO inhibitory activities than 2, 6-di-tert-butyl-4-methylphenol (BHT) or ascorbic acid, and more powerful effect on protecting erythrocytes from oxidative damage than ascorbic acid. The *OH scavenging capacity was positively proportional to the concentrations of 1 and 2 ranging from 0.015 6 to 0.500 0 mg x mL(-1). The hepatic LPO inhibitory activities increased with the increasing concentrations of 1 and 2 from 0.001 9 to 0.250 0 mg x mL(-1), but decreased slightly with the increasing concentration from 0.250 0 to 1.0000 mg x L(-1).
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[Application of Bioflex dynamic stabilization system in treating multi-segment lumbar degenerative disease].
Zhongguo Gu Shang
PUBLISHED: 05-25-2011
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To explore the value of application of Bioflex dynamic stabilization system in treating multi-segment lumbar degenerative disease.
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An epidermal stem cells niche microenvironment created by engineered human amniotic membrane.
Biomaterials
PUBLISHED: 05-18-2011
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How to amplify epidermal stem cells (ESCs) rapidly is a challenging crux in skin tissue engineering research. The present study describes the preparation of 3D micronized (300-600 ?m) amniotic membrane (mAM) by means of repeated freeze-thawing cycles to deplete cell components and homogenized with a macrohomogenizer in liquid nitrogen. This newly prepared mAM not only possessed the characteristics of a microcarrier but completely retained the basement membrane structure and abundant active substances such as NGF, HGF, KGF, bFGF, TGF-?1 and EGF in the AM matrix. The result showed that mAM combined with rotary cell culture system (RCCS) was able to amplify ESCs quickly. The relative cell viability at day 7 and 14 was significantly higher than that of the conventional 2D plate culture (326 ± 28% and 535 ± 47% versus 232 ± 21% and 307 ± 32%, P < 0.05). In addition, the new method was able to prevent cell differentiation effectively and retain the characteristics of stem cells. When mAM loaded with ESCs (ESC-mAM) was further transplanted to full-thickness skin defects in nude mice, ESCs survived well and formed a new epidermis. Four weeks after transplantation, papilla-like structures were observed, and collagen fibers were well and regularly arranged in the newly formed dermal layer. In conclusion, the mAM as a novel natural microcarrier possesses an intact basement membrane structure and bioactivities. It not only provides the microenvironment similar to the stem cell niche within the human body favorable for ex vivo culture and amplification of ESCs but can be used as the dermal scaffold in constructing a skin substitute containing ESCs for the repair of full-thickness skin defects.
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A novel and simple PCR walking method for rapid acquisition of long DNA sequence flanking a known site in microbial genome.
Mol. Biotechnol.
PUBLISHED: 05-12-2011
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Acquisition of flanking sequence adjacent to a known DNA site is an important task in microbial genome-related research. In this study, we developed a new method containing two rounds of PCR followed by cloning and sequencing. Firstly, specific primer (SP) is added into the reaction system for primary locus-specific linear amplification, and then a complex long primer (CLP) is added into the cooled reaction system for only one cycle. Amplification products from the first round of PCR are directly purified without electrophoresis, diluted, and used as the templates of the second PCR. Secondly, one long specific primer (LSP) and one long base-fixed primer (LFP) are adopted. The amplicons are purified for cloning and sequencing. The achievement of specific amplification for long flanking region mainly depends on ingenious and precise settings of PCR programs, structure design of CLP primer, adding of CLP primer after specific linear amplification, concentration ratio of CLP and SP primer, applying long primers, etc. Through this method, we successfully performed the long PCR walkings (>1.5 Kb) on rpoB gene of Vibrio vulnificus, transposon-like gene of V. alginolyticus, and sto gene of V. cholerae. The method provides a robust and simple strategy for rapid amplification of long unknown DNA fragments from microbes.
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The role of glutamate receptors in traumatic brain injury: implications for postsynaptic density in pathophysiology.
Brain Res. Bull.
PUBLISHED: 04-27-2011
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Traumatic brain injury (TBI) is the major cause of death and disability, and the incidence of TBI continues to increase rapidly. In recent years, increasing attention has been paid to an important structure at the postsynaptic membrane: the postsynaptic density (PSD). Glutamate receptors, as major components of the PSD, are highly responsive to alterations in the glutamate concentration at excitatory synapses and activate intracellular signal transduction via calcium and other second messengers following TBI. PSD scaffold proteins (PSD-95, Homer, and Shank), which anchor glutamate receptors and form a network structure, also have potential effects on these downstream signaling pathways. The changes in the function and structure of these major PSD proteins are also induced by TBI, indicating that there is a more complicated mechanism associated with PSD proteins in the pathophysiological process of TBI.
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[Application of anterior percutaneous screw fixation in treatment of odontoid process fractures in aged people].
Zhongguo Gu Shang
PUBLISHED: 04-14-2011
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To explore the efficacy of anterior percutaneous screw fixation in the treatment of odontoid process fractures in aged people.
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[Preparation and antithrombogenicity of oxidated low molecular weight heparin-antithrombin complex coated-polyvinyl chloride tubing].
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi
PUBLISHED: 04-13-2011
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Based on non-enzymatic protein glycated reaction, the sodium periodate-oxidated low molecular weight heparin-antithrombin covalent complex (SPLMWATH) was produced. By using polyethyleneimine-glutaraldehyde bonding technique, polyvinyl chloride (PVC) tubings were coated with SPLMWATH, heparin and low molecular weight heparin (LMWH). Spectrophotometry and dynamic clotting time experiment were used to determine the synthetic ratio of SPLMWATH, graft density, coating leaching ratio and to evaluate the antithrombogenicity of different coating on the PVC tubings. The results showed that the synthetic ratio of SPLMWATH was approximately 55%, and compared with heparin coating and LMWH coating, the graft density of SPLMWATH coating on the PVC tubing was smaller, but its coating stability and antithrombogenicity were significantly better than that of heparin coating and LMWH coating on the PVC tubings.
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Expression of tyrosine kinase receptor C in the segments of the spinal cord and the cerebral cortex after cord transection in adult rats.
Neurosci Bull
PUBLISHED: 03-29-2011
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To investigate the role of tyrosine kinase receptor C (TrkC), the receptor of neurotrophin-3 (NT-3), in neuroplasticity following spinal cord injury (SCI).
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[Therapeutic effects and complications of percutaneous pedicle screw fixation for thoracolumbar fractures].
Zhonghua Wai Ke Za Zhi
PUBLISHED: 03-24-2011
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To investigate the therapeutic effects and complications of percutaneous pedicle screw fixation for thoracolumbar fractures.
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A new strategy of promoting vascularization of skin substitutes by capturing endothelial progenitor cells automatically.
Med. Hypotheses
PUBLISHED: 02-22-2011
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How to promote vascularization of a skin substitute is the key to successful skin transplantation. Current methods are mainly through releasing angiogenesis-related factors (ARF) or seeding angiogenesis-related cells (ARC), but the efficacy of these methods is not satisfactory, because angiogenesis needs participation of multiple factors, extracellular matrix and related cells. The latest research has demonstrated that endothelial progenitor cells (EPCs) originating from bone marrow and existing in peripheral blood are the key element participating in revascularization of adult tissues. They directly participate in both stem cell vasculogenesis of ischemic tissues and local angiogenesis. We therefore hypothesize whether it is possible to construct a new skin substitute and use it to mobilize EPCs in bone marrow to peripheral circulation and capture EPCs automatically as a simple and effective method of promoting vascularization of the skin substitute for the sake of improving its post-transplant survival.
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SYBR Green I-based real-time PCR targeting the rpoX gene for sensitive and rapid detection of Vibrio alginolyticus.
Mol. Cell. Probes
PUBLISHED: 02-22-2011
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rpoX, a Vibrio alginolyticus specific stress regulating gene, was used to detect this fish pathogen by SYBR Green I-based real-time PCR. The specificity of the detection was confirmed in different samples. The minimum level of detection was 10(3) cells from pure culture and 10(2) cells from seawater.
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[Effect of hydrogen-rich saline on blood pressure and antioxidant ability of lung tissue in scalded rats following delayed resuscitation].
Zhonghua Shao Shang Za Zhi
PUBLISHED: 11-13-2010
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To study the effect of hydrogen-rich saline on blood pressure and antioxidant ability of lung tissue in scalded rats following delayed resuscitation.
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Permanent Genetic Resources added to Molecular Ecology Resources Database 1 April 2010 - 31 May 2010.
, K Andree, Jan Axtner, M J Bagley, E J Barlow, T J C Beebee, Jeffrey L Bennetzen, Eldredge Bermingham, M C Boisselier-Dubayle, Christine A Bozarth, Christopher P Brooks, R P Brown, Gaetano Catanese, S Cavers, Ivania Cerón-Souza, Solomon T C Chak, M N Chan, P Charles-Dominique, C Y Chen, J D Chen, Leah Chinchilla, D DA Silva, S Dafreville, F Daunt, H Delatte, T Dorge, N Duncan, J D Durand, D Duvernell, Matt Estep, Sigang Fan, R Fattahi, Oscar Flores Villela, Yokking Fong, H Fréville, Victoria Funes, C Gallardo-Escarate, K N Ganeshaiah, M R Ghaffari, C Girod, B J Gomez-Moliner, Gracia P Gonzalez-Porter, A Gosa, F Govers, F Guérin, Diarah Guindo, Frank Hailer, P A Haye, Kim A Hoelmer, S Hofmann, Yan Hong, Chaoqun Hu, S W Huang, L Humeau, Carlos Infante, S A Jackson, E Jacobsen, A Jowkar, M Kafi, M J Kermani, Hyojoong Kim, Kyung Seok Kim, Min-Young Kim, W Knibb, Ousmane A Koita, H Korpelainen, J Lambourdiere, Eloisa Lasso, R Leblois, Hang Lee, Seunghwan Lee, F C C Leung, Kenneth M Y Leung, Chunhong Li, Y Li, Dietmar Lieckfeldt, M Lizana, W J Loughry, Peng Luo, M J Madeira, P Mahmoodi, Jesus E Maldonado, M Mardi, O Mendes, G Miehe, Peter Muth, D Nacci, L Naveen Kumar, Wai-Chuen Ng, T Pailler, Heiko K Parzies, Laura Perez, M Pfunder, M Pietiläinen, S M Pirseyedi, D Porta, J Porta, J M Porta, S Quilici, F P Rakotoarivelo, B T Ramesha, G Ravikanth, B Riéra, A M Risterucci, D A Roberts, S Samadi, V Sarasola-Puente, E Sarrazin, C Sarthou, Anke Schmidt, N I Segovia, K N Shen, C Simiand, Muhammad Hidayat Bin Sman, T Solhoy, Simone Sommer, R C Sumangala, Ramona Taubert, T Tejangkura, A Telford, A Testa, C Tollon-Cordet, W N Tzeng, R Uma Shaanker, T A J Van Der Lee, Thomas A VAN Mourik, R Vasudeva, T C Wai, R L Wang, Mark E Welch, Eva Weltzien, A Whitehead, Anastasia Woodard, Jianjun Xia, M Zeinolabedini, Lvping Zhang.
Mol Ecol Resour
PUBLISHED: 09-26-2010
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This article documents the addition of 396 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Anthocidaris crassispina, Aphis glycines, Argyrosomus regius, Astrocaryum sciophilum, Dasypus novemcinctus, Delomys sublineatus, Dermatemys mawii, Fundulus heteroclitus, Homalaspis plana, Jumellea rossii, Khaya senegalensis, Mugil cephalus, Neoceratitis cyanescens, Phalacrocorax aristotelis, Phytophthora infestans, Piper cordulatum, Pterocarpus indicus, Rana dalmatina, Rosa pulverulenta, Saxifraga oppositifolia, Scomber colias, Semecarpus kathalekanensis, Stichopus monotuberculatus, Striga hermonthica, Tarentola boettgeri and Thermophis baileyi. These loci were cross-tested on the following species: Aphis gossypii, Sooretamys angouya, Euryoryzomys russatus, Fundulus notatus, Fundulus olivaceus, Fundulus catenatus, Fundulus majalis, Jumellea fragrans, Jumellea triquetra Jumellea recta, Jumellea stenophylla, Liza richardsonii, Piper marginatum, Piper aequale, Piper darienensis, Piper dilatatum, Rana temporaria, Rana iberica, Rana pyrenaica, Semecarpus anacardium, Semecarpus auriculata, Semecarpus travancorica, Spondias acuminata, Holigarna grahamii, Holigarna beddomii, Mangifera indica, Anacardium occidentale, Tarentola delalandii, Tarentola caboverdianus and Thermophis zhaoermii.
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Anti-DNA antibody modified coronary stent for plasmid gene delivery: results obtained from a porcine coronary stent model.
J Gene Med
PUBLISHED: 07-05-2010
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Previous work in our laboratory has demonstrated that the anti-DNA antibody-immobilized stent results in highly site-specific gene delivery in a rabbit carotid model. As a result of the similarity in the anatomy and physiology of the pig and human cardiovascular systems, the porcine coronary stent model was used in the present study to evaluate the site-specificity, efficiency and long-term therapeutic effect of this gene delivery system in pig coronary arteries.
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Autophagy is induced by the type III secretion system of Vibrio alginolyticus in several mammalian cell lines.
Arch. Microbiol.
PUBLISHED: 06-28-2010
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Vibrio alginolyticus is a gram-negative bacterium and has been recognized as an opportunistic pathogen in marine animals as well as humans. Here, we further characterized a cell death mechanism caused by this bacterium in several mammalian cell lines. The T3SS of V. alginolyticus killed HeLa cells by a very similar cell cytolysis mechanism in fish cells, as evidenced by cell rounding and LDH release; however, DNA fragmentation was not observed. Further studies showed that caspase-1 and caspase-3 were not activated during the T3SS-mediated cell death, indicating that the death mechanism is completely independent of pyroptosis and apoptosis in HeLa cells. Conversely, autophagy was detected during the T3SS-mediated cell death by the appearance of MDC-labeled punctate fluorescence and accumulation of autophagic vesicles. Moreover, western blot analysis revealed increase in conversion of LC3-I to LC3-II in infected mammalian cell lines, confirming that autophagy occurs during the process. Together, these data demonstrate that the death process used by V. alginolyticus in mammalian cells is different from that in fish cells, including induction of autophagy, cell rounding and osmotic lysis. This study provides some evidences hinting that differences in death mechanism in responses to V. alginolyticus infection may be attributed to the species of infected cells from which it was derived.
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The type III secretion system of Vibrio alginolyticus induces rapid apoptosis, cell rounding and osmotic lysis of fish cells.
Microbiology (Reading, Engl.)
PUBLISHED: 06-24-2010
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Vibrio alginolyticus is a Gram-negative bacterium and has been recognized as an opportunistic pathogen in humans as well as marine animals. However, the virulence mechanisms for this species of Vibrio have not been elucidated. This study characterized multiple mechanisms that induce cell death in fish cells upon infection with a V. alginolyticus strain, ZJO. The bacterium required its type III secretion system (T3SS) to cause rapid death of infected fish cells. Dying cells exhibited some features of apoptotic cells, such as membrane blebbing, nuclear condensation and DNA fragmentation. Further studies showed that caspase-3 was activated by the T3SS of the ZJO strain, confirming that infection with V. alginolyticus rapidly induces T3SS-dependent apoptosis in fish cells. Infection with the ZJO strain also led to membrane pore formation and release of cellular contents from infected fish cells, as evidenced by lactate dehydrogenase release and the uptake of a membrane-impermeable dye. Importantly, inhibition of apoptosis did not prevent ZJO-infected cells from releasing cellular contents and did not block cell rounding. Taken together, these data demonstrate that infection with V. alginolyticus may promote at least three different T3SS-dependent events, which lead to the death of fish cells. This study provides an important insight into the mechanism used by Vibrio species to cause host-cell death.
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[Establishment of loop-mediated isothermal amplification method for detection of Legionella pneumophila].
Zhejiang Da Xue Xue Bao Yi Xue Ban
PUBLISHED: 06-15-2010
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To establish a simple and rapid molecular detection for Legionella pneumophila.
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[Complete nucleotide sequence of a plasmid pVAE259 from vibrio alginolyticus and analysis of molecular biological characteristic of the plasmid].
Wei Sheng Wu Xue Bao
PUBLISHED: 04-15-2010
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To obtain the complete nucleotide sequence of the plasmid pVAE259 from Vibrio alginolyticus, to analyze molecular characteristic of the plasmid, and to explore its potential function.
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Detection of beta-glucans using an amperometric biosensor based on high-affinity interaction between Dectin-1 and beta-glucans.
Anal. Biochem.
PUBLISHED: 04-08-2010
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Early diagnosis of fungal infection plays an important role in increasing antifungal therapeutic response, but meaningful tests such as microbiological cultures and histopathological diagnosis are usually insensitive and time-consuming. A sensitive amperometric biosensor for beta-glucans was fabricated by immobilizing Dectin-1 onto Nafion-thionine-gold nanoparticle-chitosan multilayer films to trap its corresponding ligand from sample solution. On formation of ligand-receptor complex, detection of beta-glucans was accomplished by monitoring the decrease of the electrochemical signal of the modified electrode due to the inhibition of the transmission of electrons. Dectin-1 was constructed by cloning the extracellular carbohydrate recognition domain of the mouse Dectin gene into the pET28a(+) prokaryotic expression vector. Optimal conditions and analytical performances of the described biosensor were investigated. Under the optimal conditions, the biosensor response for beta-glucans presented good accuracy, stability, and reproducibility. The proposed biosensor not only could be used for rapid analysis of serum beta-glucans but also provided a screening procedure for the determination of fungal infections.
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Electrode potential regulates cytochrome accumulation on Shewanella oneidensis cell surface and the consequence to bioelectrocatalytic current generation.
Biosens Bioelectron
PUBLISHED: 03-02-2010
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This study investigated the physiological response of Shewanella oneidensis to the electrode potential and its effect on biological current generation. The electrode-attached S. oneidensis cells expressed cytochromes OmcA/MtrC at the bacteria-electrode interface when the electrode was poised at 0V (vs. SCE), but not when the poised potential was -0.24V. Cytochromes that had already been deposited on the interface were eliminated under -0.24V. Unexpectedly, current density under the higher poised potential was very low, while the low current density was associated with accumulation of OmcA/MtrC at the Shewanella-electrode interface. Although OmcA/MtrC were widely believed to be electrode reductases, we found that their electrocatalytic activity was actually weak. An additional electron transfer pathway through diffusive electron shuttle flavins was more efficient. The electrocatalytically inactive OmcA/MtrC could probably insulate the electrode surface and hinder the contact between flavins and electrode, which inhibited current generation. The results highlighted that the bacterial exoelectrogenic physiology is directly relevant to the operation of bioelectrochemical systems.
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NSAID sulindac and its analog bind RXRalpha and inhibit RXRalpha-dependent AKT signaling.
Cancer Cell
PUBLISHED: 02-21-2010
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Nonsteroidal anti-inflammatory drugs (NSAIDs) exert their anticancer effects through cyclooxygenase-2 (COX-2)-dependent and independent mechanisms. Here, we report that Sulindac, an NSAID, induces apoptosis by binding to retinoid X receptor-alpha (RXRalpha). We identified an N-terminally truncated RXRalpha (tRXRalpha) in several cancer cell lines and primary tumors, which interacted with the p85alpha subunit of phosphatidylinositol-3-OH kinase (PI3K). Tumor necrosis factor-alpha (TNFalpha) promoted tRXRalpha interaction with the p85alpha, activating PI3K/AKT signaling. When combined with TNFalpha, Sulindac inhibited TNFalpha-induced tRXRalpha/p85alpha interaction, leading to activation of the death receptor-mediated apoptotic pathway. We designed and synthesized a Sulindac analog K-80003, which has increased affinity to RXRalpha but lacks COX inhibitory activity. K-80003 displayed enhanced efficacy in inhibiting tRXRalpha-dependent AKT activation and tRXRalpha tumor growth in animals.
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C/EBP? down-regulation is associated with reduced hepatic cellular viability during hypoxia in vitro and in vivo.
Exp. Toxicol. Pathol.
PUBLISHED: 02-05-2010
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C/EBP? transcription factor is a key regulator in liver biology and was preliminarily shown to be down-regulated in hypoxic primary rat hepatocytes. The aim of this study was to explore the possible association between C/EBP? expression level and hepatocyte viability in both the in-vitro cultured hypoxic rat primary hepatocytes and two models of acute liver hypoxia induced by carbon tetrachloride or Fas antibody. C/EBP? mRNA was significantly down-regulated under hypoxic conditions both in vitro and in vivo, which was paralleled by a similar decrease in hepatocyte viability and partially reversed by 3D matrix and dexamethasone. These results suggested that C/EBP? down-regulation may be one mechanism of reduced hepatocyte viability in these settings.
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Biliary intervention aggravates cholestatic liver injury, and induces hepatic inflammation, proliferation and fibrogenesis in BDL mice.
Exp. Toxicol. Pathol.
PUBLISHED: 01-21-2010
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Obstructive cholestasis occurs in various clinical situations, whose pathological process is complex and not well known. The present study was initiated to display the complex and multifaceted pathological process caused by obstructive cholestasis in bile duct-ligated mice. Adult mice were bile-duct-ligated or sham-operated, and serum and liver tissues were collected at the indicated time points. Automatic biochemical analyzer was used to monitor serum biochemical index; TUNEL, HE staining, immunohistochemistry and Real-time PCR were employed to evaluate liver apoptosis, necrosis, inflammation, as well as proliferation and fibrosis. Our results demonstrated that obstructive cholestasis led to elevated serum biochemical indicators, with ALT peaking at day 3, indicative of acute hepatic dysfunction. Meanwhile, the number of TUNEL-positive cells increased significantly, and by 2 weeks, mild to moderate necrosis became apparent in BDL mouse livers, which consequently aggravated hepatic inflammatory responses as was demonstrated by increased expression of KC-1, MIP-2, ICAM-1 and MPO in BDL mouse livers. Moreover, proliferative hepatocytes around periportal areas, manifested by enhanced cell mitosis and elevated expression of proliferative markers such as PCNA and Ki67, increased significantly after BDL, while increased CK-19-positive cells in bile ducts indicated bile duct hyperplasia. By 2 weeks, numerous ?-SMA-positive cells and Sirius-stained collagen were observed, indicative of hepatic stellate cells (HSC) activation and fibrogenesis. In conclusion, biliary intervention led to a multifaceted hepatic pathological process characterized by aggravated liver injury and inflammatory reaction with enhanced cellular proliferation and fibrogenesis.
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