JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
Bidirectional reflectance distribution function based surface modeling of non-Lambertian using intensity data of light detection and ranging.
J Opt Soc Am A Opt Image Sci Vis
PUBLISHED: 11-18-2014
Show Abstract
Hide Abstract
To provide a credible model for light detection and ranging (LiDAR) target classification, the focus of this study is on the relationship between intensity data of LiDAR and the bidirectional reflectance distribution function (BRDF). An integration method based on the built-in-lab coaxial laser detection system was advanced. A kind of intermediary BRDF model advanced by Schlick was introduced into the integration method, considering diffuse and specular backscattering characteristics of the surface. A group of measurement campaigns were carried out to investigate the influence of the incident angle and detection range on the measured intensity data. Two extracted parameters r and S? are influenced by different surface features, which illustrate the surface features of the distribution and magnitude of reflected energy, respectively. The combination of two parameters can be used to describe the surface characteristics for target classification in a more plausible way.
Related JoVE Video
Metabolic profiling analysis of berberine, palmatine, jatrorrhizine, coptisine and epiberberine in zebrafish by ultra-high performance liquid chromatography coupled with LTQ Orbitrap mass spectrometer.
Xenobiotica
PUBLISHED: 11-06-2014
Show Abstract
Hide Abstract
Abstract 1.? Zebrafish has been used in metabolic study of drugs as a powerful tool in recent years. In this study, we make a feasible metabolism investigation of five protoberberine alkaloids (PBAs) applied in zebrafish model for the first time, including berberine (BBR), palmatine (PAL), jatrorrhizine (JAT), coptisine (COP) and epiberberine (EBBR). 2.? After exposure for 24 hours, 19 metabolites were identified by LTQ Orbitrap mass spectrometer, including 9 phase I metabolites and 10 phase II metabolites. Demethylation, hydroxylation, sulfation and glucuronidation were the major metabolic transformation of PBAs in zebrafish, which were similar to mammals. Compared with reported literatures, BBR and JAT showed high consistency between human and zebrafish in metabolic pathways. 3.? To our knowledge, this is the first time to study in vivo metabolism of COP, which provides useful information to other researchers. 4.? This study indicated that zebrafish model is feasible and reasonable to predict the metabolism of PBAs. It showed great potential for developing a novel and rapid method for predicting the metabolism of trace compounds of botanical drugs, with the advantages of lower cost, higher performance and easier set up.
Related JoVE Video
Single nucleotide polymorphisms in the D-loop region of mitochondrial DNA is associated with the kidney survival time in chronic kidney disease patients.
Ren Fail
PUBLISHED: 11-04-2014
Show Abstract
Hide Abstract
Abstract Background: The mitochondrial displacement loop (D-loop) is known to accumulate mutations and SNPs at a higher frequency than other regions of mitochondrial DNA (mtDNA). We had identified chronic kidney disease (CKD) risk-associated SNPs in the D-loop of CKD patients previously. In this study, we investigated the association of SNPs in the D-loop of mtDNA with the kidney survival of CKD. Methods: The D-loop region of mtDNA was sequenced for 119 CKD patients from the inpatient of the Fourth Hospital of Hebei Medical University. The Kaplan-Meier method was used to identify disease outcome-associated SNPs in the D-loop of CKD patients. The Cox proportional hazards model was used to identify risk factors for the kidney survival of CKD. Results: In the present study, we identified 20 SNPs with a frequency higher than 5% and assessed the relationship of these SNPs with kidney survival time in CKD patients, a SNP of 146 was identified by log-rank test for statistically significant prediction of the kidney survival time. In an overall multivariate analysis, allele 146 was identified as an independent predictor of kidney survival time in CKD patients. The survival time of kidney in the CKD patients with 146C was significantly shorter than that of kidney in CKD patients with 146T (relative risk, 2.336; 95% CI, 1.319-3.923; p?=?0.001). Conclusion: SNPs in the D-loop can predict the kidney survival of CKD patients. Analysis of genetic polymorphisms in the mitochondrial D-loop can help to identify CKD patient subgroup at high risk of a poor disease outcome.
Related JoVE Video
Metal-to-Metal Electron Transfer in Co/Fe Prussian Blue Molecular Analogues: The Ultimate Miniaturization.
J. Am. Chem. Soc.
PUBLISHED: 10-28-2014
Show Abstract
Hide Abstract
Co/Fe Prussian Blue analogues are known to display both thermally and light induced electron transfer attributed to the switching between diamagnetic {Fe(II)LS(?-CN)Co(III)LS} and paramagnetic {Fe(III)LS(?-CN)Co(II)HS} pairs (LS = low spin; HS = high spin). In this work, a dinuclear cyanido-bridged Co/Fe complex, the smallest {Fe(?-CN)Co} moiety at the origin of the remarkable physical properties of these systems, has been designed by a rational building-block approach. Combined structural, spectroscopic, magnetic and photomagnetic studies reveal that a metal-to-metal electron transfer that can be triggered in solid state by light, temperature and solvent contents, is observed for the first time in a dinuclear complex.
Related JoVE Video
Unraveling the toxicity mechanisms of the herbicide diclofop-methyl in rice: modulation of the activity of key enzymes involved in citrate metabolism and induction of cell membrane anion channels.
J. Agric. Food Chem.
PUBLISHED: 10-21-2014
Show Abstract
Hide Abstract
Residual soil concentrations of the herbicide diclofop-methyl (DM) can be toxic to other nontarget plant species, but the toxicity mechanisms at play are not fully understood. In the present study, we analyzed the toxic effect of DM on root growth and metabolism in the rice species Oryza sativa. The results show that a 48-h exposure to a trace level (5 ?g/L) of DM inhibits rice root growth by almost 70%. A 48-h exposure to 5 ?g/L DM also leads to an ?2.5-fold increase in citrate synthase (CS) activity (and CS gene transcription) and an ?2-fold decrease in the citrate lyase gene transcripts, which lead to an increase in the intracellular concentration of citrate and in citrate exudation rate. Addition of a specific inhibitor of cell membrane anion channel, anthracene-9-carboxylic acid, decreased citrate release in the culture, suggesting that DM-induced citrate loss from the cells is mediated by a specific membrane-bound channel protein. This study brings new insights into the key biochemical mechanisms leading to DM toxicity in rice.
Related JoVE Video
Projective rectification of infrared images from air-cooled condenser temperature measurement by using projection profile features and cross-ratio invariability.
Appl Opt
PUBLISHED: 10-17-2014
Show Abstract
Hide Abstract
In this paper, we propose a projective rectification method for infrared images obtained from the measurement of temperature distribution on an air-cooled condenser (ACC) surface by using projection profile features and cross-ratio invariability. In the research, the infrared (IR) images acquired by the four IR cameras utilized are distorted to different degrees. To rectify the distorted IR images, the sizes of the acquired images are first enlarged by means of bicubic interpolation. Then, uniformly distributed control points are extracted in the enlarged images by constructing quadrangles with detected vertical lines and detected or constructed horizontal lines. The corresponding control points in the anticipated undistorted IR images are extracted by using projection profile features and cross-ratio invariability. Finally, a third-order polynomial rectification model is established and the coefficients of the model are computed with the mapping relationship between the control points in the distorted and anticipated undistorted images. Experimental results obtained from an industrial ACC unit show that the proposed method performs much better than any previous method we have adopted. Furthermore, all rectified images are stitched together to obtain a complete image of the whole ACC surface with a much higher spatial resolution than that obtained by using a single camera, which is not only useful but also necessary for more accurate and comprehensive analysis of ACC performance and more reliable optimization of ACC operations.
Related JoVE Video
Serological Characterization of Surface-Exposed Proteins of Coxiella burnetii.
Microbiology (Reading, Engl.)
PUBLISHED: 10-10-2014
Show Abstract
Hide Abstract
The obligate intracellular Gram-negative bacterium Coxiella burnetii causes Q fever, a worldwide zoonosis. Here we labeled C. burnetii with biotin and used biotin-streptavidin affinity chromatography to isolate surface-exposed proteins (SEPs). Using two-dimensional electrophoresis combined with mass spectrometry, we identified 37 proteins through bioinformatics analysis. Thirty SEPs expressed in Escherichia coli (recombinant SEPs, rSEPs) were used to generate microarrays, which were probed with sera from mice experimentally infected with C. burnetii or sera from Q fever patients. Thirteen rSEPs were recognized as seroreactive, and the majority reacted with at least 50% of the sera from mice infected with C. burnetii but not with sera from mice infected with Rickettsia rickettsii, R. heilongjiangensis, or R. typhi. Further, 13 proteins that reacted with sera from patients with Q fever did not react with sera from patients with brucellosis or mycoplasma pneumonia. Our results suggest that these seroreactive SEPs have potential as serodiagnostic antigens or as subunit vaccine antigens against Q fever.
Related JoVE Video
A meta-analysis on the relationship of eNOS 4b/a polymorphism and diabetic nephropathy susceptibility.
Ren Fail
PUBLISHED: 10-08-2014
Show Abstract
Hide Abstract
Abstract To clarify the effect of nitric oxide synthase (NOS) type III 4b/a polymorphism on the susceptibility to diabetic nephropathy (DN) by meta-analysis, we performed a computerized search of PubMed, EMBASE, Chinese BioMedical Literature Database, China Science and Technology Journal Database, Chinese Journal Full-text Database and WanFang to identity case-control studies on relationship between NOS type III 4b/a polymorphism and the susceptibility to DN. Statistic analysis and heterogeneity test were conducted by StataSE12. The meta-analysis involved 26 studies for DN comparing with diabetes mellitus (DM) and 15 studies for DN comparing with healthy persons, which provided 6144/4900 cases/controls and 2134/2348 cases/controls, respectively. Moderate heterogeneity was found among including studies. The qualities of half studies are low. Meta-analysis derived a significant association between the NOS type III 4b/a and the risk of developing DN in Asian population. The sensitivity analysis (exclusion of studies not in Hardy-Weinberg equilibrium) produced non-significant changes. Compared with diabetes patients, the pre-allele model produced certain association in global populations [odds ratio (OR)?=?1.26, 95% confidence interval (95% CI): 1.10-1.45], significant association in Asian population (OR?=?1.51, 95% CI: 1.13-2.01) and certain association in type 2 DM patients (OR?=?1.29, 95% CI: 1.09-1.54). Only in the dominant model, the funnel plot and Egger's test provided evidence of publication bias (p?=?0.024). Overall, although there is some evidence of association between NOS type III 4b/a polymorphism and DN in Asian population, the more reliable findings need further and more rigorous, prospective and high-quality studies.
Related JoVE Video
A randomized trial in healthy subjects to assess the bioequivalence of an atazanavir/cobicistat fixed-dose combination tablet versus administration as separate agents.
Antivir. Ther. (Lond.)
PUBLISHED: 10-05-2014
Show Abstract
Hide Abstract
Cobicistat (COBI) is an alternative pharmacoenhancer to ritonavir. A fixed-dose combination (FDC) tablet containing atazanavir (ATV) and COBI is under development for the treatment of HIV-1-infected patients.
Related JoVE Video
An Avian Leukosis Virus Subgroup J Isolate with Rous Sarcoma Virus-like 5'-LTR shows enhanced replication capability.
J. Gen. Virol.
PUBLISHED: 10-03-2014
Show Abstract
Hide Abstract
Avian leukosis virus subgroup J (ALV-J) was first isolated from meat-type chickens that had developed myeloid leukosis. However, ALV-J infections associated with hemangiomas have occurred in layer flock in China. In this study, we identified an ALV-J layer isolate (HLJ13SH01) as a recombinant of ALV-J and a Rous sarcoma virus Schmidt Ruppin B strain (RSV-SRB), which contained the RSV-SRB 5'-LTR region and the other genes of ALV-J. Replication kinetic testing indicated that the HLJ13SH01 strain replicated faster than other ALV-J layer isolates in vitro. Sequence analysis indicated that the main difference between the two isolates was the 5'-LTR sequences, particularly the U3 sequences. A 19-nt insertion was uniquely found in the U3 region of the HLJ13SH01 strain. The results of the Dual-Glo Luciferase revealed that the 19-nt insertion in the HLJ13SH01 strain increased the enhancer activity of the U3 region. Moreover, an additional CCAAT/enhancer element was found in the 19-nt insertion, and the luciferase assay indicated that this element played a key role in increasing the enhancer activity of the 5'-U3 region. To confirm the potentiation effect of the 19-nt insertion and the CCAAT/enhancer element on virus replication, three infectious clones with 5'-U3 region variations were constructed and rescued. Replication kinetic testing of the rescued viruses demonstrated that the CCAAT/enhancer element in the 19-nt insertion enhanced the replication capacity of the ALV-J recombinant in vitro.
Related JoVE Video
Uniform core-shell photonic crystal microbeads as microcarriers for optical encoding.
Langmuir
PUBLISHED: 10-01-2014
Show Abstract
Hide Abstract
We demonstrate a rapid and robust method to fabricate uniform core-shell photonic crystal (PC) microbeads by the microfluidic and centrifugation-redispersion technique. Colored crystalline colloidal arrays (CCAs) were first prepared through centrifugation-redispersion approach by self-assembly of polystyrene-poly(N-isopropylacrylamide) (PS-PNIPAm) core/shell nanoparticles (NPs). Different from the conventional NPs (e.g., charged PS or PNIPAm NPs), PS-PNIPAm NPs could easily self-assemble into well-ordered CCAs by only one purification step without laborious pretreatment (e.g., dialysis or ion exchange) or slow solvent-evaporation process. The CCAs is then encapsulated into a transparent polymer shell with functional groups (e.g., copolymer of ETPTA and butyl acrylate (BA)), triggering the formation of core-shell PC microbeads which can be used as optical encoding microcarriers. Importantly, this technique allows us to produce core-shell PC microbeads in a rapid and robust way, and the optical reflections of the PC microbeads appear highly stable to various external stimuli (e.g., temperature, pH value, and ionic strength) relying on the features of the CCAs core and protection of the polymer shell. Moreover, various probe biomolecules (e.g., proteins, antibodies, and so on) can be easily linked on the surface of the core-shell PC microbeads owing to the hydrophilic modification induced by the hydrolysis of BA on the microbead surface, enabling the multiplex biomolecular detection.
Related JoVE Video
Predictors of poor sleep quality and excessive daytime sleepiness in peritoneal dialysis patients.
Ren Fail
PUBLISHED: 09-17-2014
Show Abstract
Hide Abstract
Abstract To explore the possible impact factors on daytime sleepiness among peritoneal patients from a single center in China. A cross-sectional study was conducted in 98 prevalent peritoneal dialysis (PD) patients using both the Pittsburgh Sleep Quality Index (PSQI) questionnaire of sleep quality and the Epworth Sleepiness Scale (ESS) questionnaire of excessive daytime sleepiness (EDS). Biochemical differences between daytime sleepiness and non-daytime sleepiness population were evaluated, following univariate and multivariable analysis to find the risk factors on sleep disturbance. The prevalence of "poor sleep quality" (PSQI?>?5) was 74.49%, while daytime sleepiness (ESS???9) occurred in 22.45%. Mean PSQI was 9.06?±?4.60 and EES was 6.31?±?4.98. Compared to non-EDS cases, patients with ESS???9 had worse residual renal function (RRF), higher serum creatinine, higher serum magnesium and elevated serum ferritin. In univariate analysis, ESS correlated with serum albumin (r?=?0.346, p?=?0.015), phosphate (r?=?0.313, p?=?0.029), magnesium (r?=?0.376, p?=?0.008) and urinary Kt/V (r?=?-0.341, p?=?0.029). Finally, multivariable linear regression indicated that urinary Kt/V, PSQI and magnesium were independent predictors of ESS score. EDS does exist in PD patients and is associated both with poor nighttime sleep quality and lower RRF. Hypermagnesemia may be a treatable risk factor to improve daytime tiredness.
Related JoVE Video
[Non-linear research of alertness levels under sleep deprivation].
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi
PUBLISHED: 09-16-2014
Show Abstract
Hide Abstract
We applied Lempel-Ziv complexity (LZC) combined with brain electrical activity mapping (BEAM) to study the change of alertness under sleep deprivation in our research. Ten subjects were involved in 36 hours sleep deprivation (SD), during which spontaneous electroencephalogram (EEG) experiments and auditory evoked EEG experiments-Oddball were recorded once every 6 hours. Spontaneous and evoked EEG data were calculated and BEAMs were structured. Results showed that during the 36 hours of SD, alertness could be divided into three stages, i. e. the first 12 hours as the high stage, the middle 12 hours as the rapid decline stage and the last 12 hours as the low stage. During the period SD, LZC of Spontaneous EEG decreased over the whole brain to some extent, but remained consistent with the subjective scales. By BEAMs of event related potential, LZC on frontal cortex decreased, but kept consistent with the behavioral responses. Therefore, LZC can be effective to reflect the change of brain alertness. At the same time LZC could be used as a practical index to monitor real-time alertness because of its simple computation and fast calculation.
Related JoVE Video
Efficient anharmonic vibrational spectroscopy for large molecules using local-mode coordinates.
J Chem Phys
PUBLISHED: 09-15-2014
Show Abstract
Hide Abstract
This article presents a general computational approach for efficient simulations of anharmonic vibrational spectra in chemical systems. An automated local-mode vibrational approach is presented, which borrows techniques from localized molecular orbitals in electronic structure theory. This approach generates spatially localized vibrational modes, in contrast to the delocalization exhibited by canonical normal modes. The method is rigorously tested across a series of chemical systems, ranging from small molecules to large water clusters and a protonated dipeptide. It is interfaced with exact, grid-based approaches, as well as vibrational self-consistent field methods. Most significantly, this new set of reference coordinates exhibits a well-behaved spatial decay of mode couplings, which allows for a systematic, a priori truncation of mode couplings and increased computational efficiency. Convergence can typically be reached by including modes within only about 4 Å. The local nature of this truncation suggests particular promise for the ab initio simulation of anharmonic vibrational motion in large systems, where connection to experimental spectra is currently most challenging.
Related JoVE Video
Facile synthesis of soluble functional graphene by reduction of graphene oxide via acetylacetone and its adsorption of heavy metal ions.
Nanotechnology
PUBLISHED: 09-11-2014
Show Abstract
Hide Abstract
The synthesis of graphene (GR) from graphene oxide (GO) typically involves harmful chemical reducing agents that are undesirable for most practical applications. Here we report a green and facile synthesis method for the synthesis of GR that is soluble in water and organic solvents and that includes the additional benefit of adsorption of heavy metal ions. Acetylacetone, as both a reducing agent and a stabilizer, was used to prepare soluble GR from GO. Transmission electron microscopy and atomic force microscopy provide clear evidence for the formation of few-layer GR. The results from Fourier transform infrared spectroscopy and ultraviolet-visible spectroscopy show that reduction of GO to GR has occurred. Raman spectroscopy and X-ray photoelectron spectroscopy also indicate the removal of oxygen-containing functional groups from GO, resulting in the formation of GR. The results of dispersion experiments show that GR can be highly dispersed in water and N,N-Dimethylformamide. The reaction mechanism for acetylacetone reduction of exfoliated GO was also proposed. This method is a facile and environmentally friendly approach to the synthesis of GR and opens up new possibilities for preparing GR and GR-based nanomaterials for large-scale applications. Of even greater interest is that inductively coupled plasma atomic emission spectroscopy suggests that synthesized GR may be applied in the absorption of Cd(2+) and Co(2+) due to the strong coordination capacity of acetylacetone on the surfaces and edges of GR and the large surface area of GR in aqueous solutions. The maximum adsorptions are 49.28 mg g(-1) for Cd(2+), which is 4.5 times higher than that of carbon nanotubes, and 27.78 mg g(-1) for Co(2+), which is 3.6 times higher than that of titania beans.
Related JoVE Video
Single nucleotide polymorphisms in the D-loop region of mitochondrial DNA and age-at-onset of patients with chronic kidney disease.
Chin. Med. J.
PUBLISHED: 09-06-2014
Show Abstract
Hide Abstract
The mitochondrial displacement loop (D-loop) accumulates mutations and single nucleotide polymorphisms (SNPs) at a higher frequency than other regions of mitochondrial DNA (mtDNA). We previously identified disease risk-associated SNPs in the D-loop of chronic kidney disease (CKD) patients; in this study, we investigated the association of age-at-onset and D-loop SNPs in CKD patients.
Related JoVE Video
Cell-Derived Vesicles for Single-Molecule Imaging of Membrane Proteins.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 09-01-2014
Show Abstract
Hide Abstract
A new approach is presented for the application of single-molecule imaging to membrane receptors through the use of vesicles derived from cells expressing fluorescently labeled receptors. During the isolation of vesicles, receptors remain embedded in the membrane of the resultant vesicles, thus allowing these vesicles to serve as nanocontainers for single-molecule measurements. Cell-derived vesicles maintain the structural integrity of transmembrane receptors by keeping them in their physiological membrane. It was demonstrated that receptors isolated in these vesicles can be studied with solution-based fluorescence correlation spectroscopy (FCS) and can be isolated on a solid substrate for single-molecule studies. This technique was applied to determine the stoichiometry of ?3?4 nicotinic receptors. The method provides the capability to extend single-molecule studies to previously inaccessible classes of receptors.
Related JoVE Video
Tanshinon IIA injection accelerates tissue expansion by reducing the formation of the fibrous capsule.
PLoS ONE
PUBLISHED: 08-26-2014
Show Abstract
Hide Abstract
The tissue expansion technique has been applied to obtain new skin tissue to repair large defects in clinical practice. The implantation of tissue expander could initiate a host response to foreign body (FBR), which leads to fibrotic encapsulation around the expander and prolongs the period of tissue expansion. Tanshinon IIA (Tan IIA) has been shown to have anti-inflammation and immunoregulation effect. The rat tissue expansion model was used in this study to observe whether Tan IIA injection systematically could inhibit the FBR to reduce fibrous capsule formation and accelerate the process of tissue expansion. Forty-eight rats were randomly divided into the Tan IIA group and control group with 24 rats in each group. The expansion was conducted twice a week to maintain a capsule pressure of 60 mmHg. The expansion volume and expanded area were measured. The expanded tissue in the two groups was harvested, and histological staining was performed; proinflammatory cytokines such as tumor necrosis factor-? (TNF-?), interleukin-6 (IL-6) and interleukin-1? (IL-1?) and transforming growth factor-? (TGF-?) were examined. The expansion volume and the expanded area in the Tan IIA group were greater than that of the control group. The thickness of the fibrous capsule in the Tan IIA group was reduced with no influence on the normal skin regeneration. Decreased infiltration of macrophages, lower level of TNF-?, IL-6, IL-1? and TGF-?, less proliferating myofibroblasts and enhanced neovascularization were observed in the Tan IIA group. Our findings indicated that the Tan IIA injection reduced the formation of the fibrous capsule and accelerated the process of tissue expansion by inhibiting the FBR.
Related JoVE Video
Effects of Atrioventricular Nodal Ablation on Permanent Atrial Fibrillation Patients With Cardiac Resynchronization Therapy: A Systematic Review and Meta-analysis.
Clin Cardiol
PUBLISHED: 08-25-2014
Show Abstract
Hide Abstract
Cardiac resynchronization therapy (CRT) is a well-established therapy for patients with heart failure (HF) and wide QRS configuration, especially for those in sinus rhythm. However, for those with permanent AF, atrioventricular nodal (AVN) ablation use remains under debate. Our objective was to evaluate clinical outcomes and mortality of AVN ablation in HF patients with permanent AF receiving CRT. Electronic publication database and reference lists through October 1, 2013 were searched. Observational cohort studies comparing CRT patients with AF who received either AVN ablation or medical therapy were selected. Outcomes included mortality, CRT nonresponse, changes in left ventricular remodeling, and functional outcomes, such as New York Heart Association (NYHA) functional class, quality of life, and 6-minute hall walk distance. Of 1641 reports identified, 13 studies with 1256 patients were included. Among patients with permanent AF and insufficient biventricular pacing (<90%), those who had undergone AVN ablation compared to those who did not had numerically lower all-cause mortality (risk ratio [RR]: 0.63, 95% confidence interval [CI]: 0.42 to 0.96, P?=?0.03) and significantly lower nonresponse to CRT (RR: 0.41, 95% CI: 0.31 to 0.54, P?
Related JoVE Video
Aldosterone induces C-reactive protein expression via MR-ROS-MAPK-NF-?B signal pathway in rat vascular smooth muscle cells.
Mol. Cell. Endocrinol.
PUBLISHED: 08-07-2014
Show Abstract
Hide Abstract
Atherosclerosis is a chronic inflammatory disease in the vessel. As a representative inflammatory cytokine, C-reactive protein (CRP) participates in atherogenesis. Although hyperaldosteronism is known to evoke inflammatory response in several tissues and cell types, there is no direct evidence to demonstrate the proinflammatory effect of aldosterone on vascular smooth muscle cells (VSMCs) through CRP. In this study, we observed the effect of aldosterone on CRP expression and the molecular mechanisms in rat VSMCs. The results showed that aldosterone induced CRP expression in VSMCs in vitro and in vivo. Mineralocorticoid receptor (MR) antagonist spironolactone abolished aldosterone-induced CRP expression. In addition, aldosterone stimulated generation of reactive oxygen species (ROS) and activated ERK1/2 phosphorylation, whereas spironolactone inhibited aldosterone-stimulated ROS generation and ERK1/2 phosphorylation. Antioxidant NAC decreased aldosterone-induced CRP expression and ERK1/2 phosphorylation. The further study confirmed that ERK1/2 inhibitor PD98059 and NF-?B inhibitor pyrrolidine dithiocarbamate both depressed aldosterone-induced CRP expression. These demonstrate that aldosterone is able to induce CRP expression via MR-ROS-ERK1/2-NF-?B signal pathway in VSMCs, which provides a new evidence for the proinflammatory and proatherosclerotic effects of aldosterone.
Related JoVE Video
Prenatal Ethanol Exposure Up-Regulates the Cholesterol Transporters ATP-Binding Cassette A1 and G1 and Reduces Cholesterol Levels in the Developing Rat Brain.
Alcohol Alcohol.
PUBLISHED: 08-02-2014
Show Abstract
Hide Abstract
Cholesterol plays a pivotal role in many aspects of brain development; reduced cholesterol levels during brain development, as a consequence of genetic defects in cholesterol biosynthesis, leads to severe brain damage, including microcephaly and mental retardation, both of which are also hallmarks of the fetal alcohol syndrome. We had previously shown that ethanol up-regulates the levels of two cholesterol transporters, ABCA1 (ATP binding cassette-A1) and ABCG1, leading to increased cholesterol efflux and decreased cholesterol content in astrocytes in vitro. In the present study we investigated whether similar effects could be seen in vivo.
Related JoVE Video
Morphology, morphogenesis and molecular phylogeny of a soil ciliate, Pseudouroleptus caudatus caudatus Hemberger, 1985 (Ciliophora, Hypotricha), from Lhalu Wetland, Tibet.
Eur. J. Protistol.
PUBLISHED: 08-01-2014
Show Abstract
Hide Abstract
Pseudouroleptus caudatus caudatus Hemberger, 1985, a soil ciliate isolated from Tibet, was studied in vivo and after protargol impregnation. The Tibetan population is mainly characterized by: elongate body with narrowly rounded anterior end and tapered posterior end; length of buccal area relative to body length ca. 20-25%; cortical granules colourless, round, densely distributed throughout sub-pellicular layer of cell; one parabuccal cirrus; post-peristomial cirrus lacking in 75% of specimens analyzed; left and right ventral rows commence at same level; four dorsal kineties; 3-6 inconspicuous caudal cirri; two macronuclear nodules; 2-7 micronuclei; contractile vacuole located at about 33% of body length near left margin. Morphogenesis is characterized by: (1) parental adoral zone of membranelles retained completely; (2) anterior segments of streaks VI and IV and the whole of streak V form the anterior, middle, posterior segments of the mixed row, respectively; (3) right ventral row originates de novo in both daughter cells; (4) marginal rows develop intrakinetally; (5) dorsal kinety anlage 3 develops de novo in the proter and intrakinetally in the opisthe; and (6) the two macronuclear nodules fuse into a single mass which then divides. Molecular phylogenies corroborate the morphological identification and support the close relationship between Pseudouroleptus and Strongylidium.
Related JoVE Video
Variational mixed quantum/semiclassical simulation of dihalogen guest and rare-gas solid host dynamics.
J Chem Phys
PUBLISHED: 07-24-2014
Show Abstract
Hide Abstract
A variational mixed quantum-semiclassical theory for the internal nuclear dynamics of a small molecule and the induced small-amplitude coherent motion of a low-temperature host medium is developed, tested, and used to simulate the temporal evolution of nonstationary states of the internal molecular and surrounding medium degrees of freedom. In this theory, termed the Fixed Vibrational Basis/Gaussian Bath (FVB/GB) method, the system is treated fully quantum mechanically while Gaussian wave packets are used for the bath degrees of freedom. An approximate time-dependent wave function of the entire model is obtained instead of just a reduced system density matrix, so the theory enables the analysis of the entangled system and bath dynamics that ensues following initial displacement of the internal-molecular (system) coordinate from its equilibrium position. The norm- and energy-conserving properties of the propagation of our trial wave function are natural consequences of the Dirac-Frenkel-McLachlan variational principle. The variational approach also stabilizes the time evolution in comparison to the same ansatz propagated under a previously employed locally quadratic approximation to the bath potential and system-bath interaction terms in the bath-parameter equations of motion. Dynamics calculations are carried out for molecular iodine in a 2D krypton lattice that reveal both the time-course of vibrational decoherence and the details of host-atom motion accompanying energy dissipation and dephasing. This work sets the stage for the comprehensive simulation of ultrafast time-resolved optical experiments on small molecules in low-temperature solids.
Related JoVE Video
Physicochemical and chromatographic characteristics of random amphiphilic copolymer aggregation in electrokinetic chromatography.
J Chromatogr A
PUBLISHED: 07-19-2014
Show Abstract
Hide Abstract
The random amphiphilic polymeric aggregation, self-assembled from poly (methyl methacrylate-co-methacrylic acid) (P(MMA-co-MAA)), was explored as a novel pseudostationary phase (PSP) in electrokinetic chromatography (EKC) in our previous report. This work focused on physicochemical characteristics and PSP performances of the polymeric aggregations. The physicochemical characteristics of polymeric aggregations, including critical aggregation concentration (CAC), zeta potential, hydrodynamic diameter, and micropolarity were determined. Experimental results showed that polymeric aggregations had much lower CAC, which decreased the usage of copolymer in EKC, weakened ionic strength and shortened analysis time. The monomer molar ratio of the copolymer was a key factor for physicochemical characteristics and PSP performances of the polymeric aggregations. With the increase of the hydrophobic monomer molar ratio, CAC, micropolarity and dimension of polymeric aggregation decreased significantly while zeta potentials were similar. Correspondingly, separation window enlarged and methylene selectivity evaluated with six kinds of n-alkylphenone homologous series enhanced. Linear solvation energy relationships (LSER) analysis found that hydrophobic interaction is the most important interaction between analytes and polymeric PSPs. Compared with SDS micelle, polymeric aggregations owned more types of interactions, such as stronger hydrogen bonding and relative larger dipole interaction, which provided a bigger adjustment room to improve PSP selectivity.
Related JoVE Video
An evaluation and recommendation of the optimal methodologies to detect RET gene rearrangements in papillary thyroid carcinoma.
Genes Chromosomes Cancer
PUBLISHED: 07-16-2014
Show Abstract
Hide Abstract
To recommend a reliable and clinically realistic RET/PTC rearrangement detection assay for papillary thyroid carcinoma (PTC), we compared multiplex quantitative polymerase chain reaction (qPCR), fluorescence in situ hybridization (FISH), and immunohistochemistry (IHC). RET/PTC rearrangement was detected using either RET break-apart FISH followed by multicolor FISH to confirm CCDC6/RET or NCOA4/RET fusions, or by multiplex qPCR to detect 14 RET/PTC subtypes with simultaneous RET mRNA expression. RET protein expression was detected by IHC. The specificity and sensitivity of multiplex qPCR and IHC were calculated using break-apart FISH as a reference. Among 73 PTC patients with sufficient tissue available for FISH and multiplex qPCR, 10 cases were defined as RET/PTC positive by both assays, including eight CCDC6/RET and two NCOA4/RET fusions with relatively high RET mRNA. In addition, multiplex qPCR identified another two CCDC6/RET fusion positive cases, but with low RET mRNA expression. IHC staining identified 11 RET positive cases among 39 patients with available samples. In comparison to FISH, multiplex qPCR displayed 100% sensitivity and 97% specificity to detect RET/PTC fusions, while IHC was neither sensitive nor specific. Our data reveal that both multiplex qPCR and FISH assays are equally applicable for detection of RET/PTC rearrangements. Break-apart FISH methodology is highly recommended for the wider screening of RET rearrangements (regardless of partner genes), while multiplex qPCR is preferred to identify all known fusion types using one assay, provided mRNA expression is also measured. IHC analysis could potentially provide an additional method of fusion detection dependent on further optimization of assay conditions and scoring cutoffs. © 2014 Wiley Periodicals, Inc.
Related JoVE Video
Efficacy of the novel oxazolidinone compound FYL-67 for preventing biofilm formation by Staphylococcus aureus.
J. Antimicrob. Chemother.
PUBLISHED: 07-04-2014
Show Abstract
Hide Abstract
Infections of hospitalized patients caused by biofilms formed by Staphylococcus aureus represent a major problem. Using in vitro and in vivo biofilm models, we evaluated the efficacy of the novel oxazolidinone FYL-67, by using linezolid (the only clinically approved oxazolidinone antibiotic) as a control, for inhibiting S. aureus biofilm formation.
Related JoVE Video
Aromatic organosulfates in atmospheric aerosols: synthesis, characterization, and abundance.
Atmos Environ (1994)
PUBLISHED: 07-01-2014
Show Abstract
Hide Abstract
Aromatic organosulfates are identified and quantified in fine particulate matter (PM2.5) from Lahore, Pakistan, Godavari, Nepal, and Pasadena, California. To support detection and quantification, authentic standards of phenyl sulfate, benzyl sulfate, 3-and 4-methylphenyl sulfate and 2-, 3-, and 4-methylbenzyl sulfate were synthesized. Authentic standards and aerosol samples were analyzed by ultra-performance liquid chromatography (UPLC) coupled to negative electrospray ionization (ESI) quadrupole time-of-flight (ToF) mass spectrometry. Benzyl sulfate was present in all three locations at concentrations ranging from 4 - 90 pg m(-3). Phenyl sulfate, methylphenyl sulfates and methylbenzyl sulfates were observed intermittently with abundances of 4 pg m(-3), 2-31 pg m(-3), 109 pg m(-3), respectively. Characteristic fragment ions of aromatic organosulfates include the sulfite radical ((•)SO3 (-), m/z 80) and the sulfate radical ((•)SO4 (-),m/z 96). Instrumental response factors of phenyl and benzyl sulfates varied by a factor of 4.3, indicating that structurally-similar organosulfates may have significantly different instrumental responses and highlighting the need to develop authentic standards for absolute quantitation organosulfates. In an effort to better understand the sources of aromatic organosulfates to the atmosphere, chamber experiments with the precursor toluene were conducted under conditions that form biogenic organosulfates. Aromatic organosulfates were not detected in the chamber samples, suggesting that they form through different pathways, have different precursors (e.g. naphthalene or methylnaphthalene), or are emitted from primary sources.
Related JoVE Video
In vitro and in vivo activities of antimicrobial peptides developed using an amino acid-based activity prediction method.
Antimicrob. Agents Chemother.
PUBLISHED: 06-30-2014
Show Abstract
Hide Abstract
To design and discover new antimicrobial peptides (AMPs) with high levels of antimicrobial activity, a number of machine-learning methods and prediction methods have been developed. Here, we present a new prediction method that can identify novel AMPs that are highly similar in sequence to known peptides but offer improved antimicrobial activity along with lower host cytotoxicity. Using previously generated AMP amino acid substitution data, we developed an amino acid activity contribution matrix that contained an activity contribution value for each amino acid in each position of the model peptide. A series of AMPs were designed with this method. After evaluating the antimicrobial activities of these novel AMPs against both Gram-positive and Gram-negative bacterial strains, DP7 was chosen for further analysis. Compared to the parent peptide HH2, this novel AMP showed broad-spectrum, improved antimicrobial activity, and in a cytotoxicity assay it showed lower toxicity against human cells. The in vivo antimicrobial activity of DP7 was tested in a Staphylococcus aureus infection murine model. When inoculated and treated via intraperitoneal injection, DP7 reduced the bacterial load in the peritoneal lavage solution. Electron microscope imaging and the results indicated disruption of the S. aureus outer membrane by DP7. Our new prediction method can therefore be employed to identify AMPs possessing minor amino acid differences with improved antimicrobial activities, potentially increasing the therapeutic agents available to combat multidrug-resistant infections.
Related JoVE Video
Property- and structure-guided discovery of a tetrahydroindazole series of interleukin-2 inducible T-cell kinase inhibitors.
J. Med. Chem.
PUBLISHED: 06-27-2014
Show Abstract
Hide Abstract
Interleukin-2 inducible T-cell kinase (ITK), a member of the Tec family of tyrosine kinases, plays a major role in T-cell signaling downstream of the T-cell receptor (TCR), and considerable efforts have been directed toward discovery of ITK-selective inhibitors as potential treatments of inflammatory disorders such as asthma. Using a previously disclosed indazole series of inhibitors as a starting point, and using X-ray crystallography and solubility forecast index (SFI) as guides, we evolved a series of tetrahydroindazole inhibitors with improved potency, selectivity, and pharmaceutical properties. Highlights include identification of a selectivity pocket above the ligand plane, and identification of appropriate lipophilic substituents to occupy this space. This effort culminated in identification of a potent and selective ITK inhibitor (GNE-9822) with good ADME properties in preclinical species.
Related JoVE Video
Volitinib, a potent and highly selective c-Met inhibitor, effectively blocks c-Met signaling and growth in c-MET amplified gastric cancer patient-derived tumor xenograft models.
Mol Oncol
PUBLISHED: 06-11-2014
Show Abstract
Hide Abstract
To investigate the incidence of cMET gene copy number changes and protein overexpression in Chinese gastric cancer (GC) and to preclinically test the hypothesis that the novel, potent and selective cMET small-molecule inhibitor volitinib, will deliver potent anti-tumor activity in cMET-dysregulated GC patient-derived tumor xenograft (PDX) models.
Related JoVE Video
Mutational analysis of dimeric linkers in peri- and cytoplasmic domains of histidine kinase DctB reveals their functional roles in signal transduction.
Open Biol
PUBLISHED: 06-06-2014
Show Abstract
Hide Abstract
Membrane-associated histidine kinases (HKs) in two-component systems respond to environmental stimuli by autophosphorylation and phospho-transfer. HK typically contains a periplasmic sensor domain that regulates the cytoplasmic kinase domain through a conserved cytoplasmic linker. How signal is transduced from the ligand-binding site across the membrane barrier remains unclear. Here, we analyse two linker regions of a typical HK, DctB. One region connects the first transmembrane helix with the periplasmic Per-ARNT-Sim domains, while the other one connects the second transmembrane helix with the cytoplasmic kinase domains. We identify a leucine residue in the first linker region to be essential for the signal transduction and for maintaining the delicate balance of the dimeric interface, which is key to its activities. We also show that the other linker, belonging to the S-helix coiled-coil family, plays essential roles in signal transduction inside the cell. Furthermore, by combining mutations with opposing activities in the two regions, we show that these two signalling transduction elements are integrated to produce a combined effect on the final activity of DctB.
Related JoVE Video
Orthologs of human disease associated genes and RNAi analysis of silencing insulin receptor gene in Bombyx mori.
Int J Mol Sci
PUBLISHED: 06-01-2014
Show Abstract
Hide Abstract
The silkworm, Bombyx mori L., is an important economic insect that has been domesticated for thousands of years to produce silk. It is our great interest to investigate the possibility of developing the B. mori as human disease model. We searched the orthologs of human disease associated genes in the B. mori by bi-directional best hits of BLAST and confirmed by searching the OrthoDB. In total, 5006 genes corresponding to 1612 kinds of human diseases had orthologs in the B. mori, among which, there are 25 genes associated with diabetes mellitus. Of these, we selected the insulin receptor gene of the B. mori (Bm-INSR) to study its expression in different tissues and at different developmental stages and tissues. Quantitative PCR showed that Bm-INSR was highly expressed in the Malpighian tubules but expressed at low levels in the testis. It was highly expressed in the 3rd and 4th instar larvae, and adult. We knocked down Bm-INSR expression using RNA interference. The abundance of Bm-INSR transcripts were dramatically reduced to ~4% of the control level at 6 days after dsRNA injection and the RNAi-treated B. mori individuals showed apparent growth inhibition and malformation such as abnormal body color in black, which is the typical symptom of diabetic patients. Our results demonstrate that B. mori has potential use as an animal model for diabetic mellitus research.
Related JoVE Video
Paeonol pretreatment attenuates cerebral ischemic injury via upregulating expression of pAkt, Nrf2, HO-1 and ameliorating BBB permeability in mice.
Brain Res. Bull.
PUBLISHED: 05-16-2014
Show Abstract
Hide Abstract
Oxidative damage plays a pivotal role in the pathogenesis of cerebral ischemic stroke and may represent a target for treatment. Our previous studies have proved that nuclear factor E2-related factor 2 (Nrf2) and its downstream genes served as a key mechanism for protection against oxidative stress. Paeonol (PN) is reputed to possess a broad range of therapeutic properties probably by virtue of its antioxidative ability. However little is elucidated regarding the underlying mechanisms in ischemic stroke. The aim of this study was to explore PNs effect in ischemic injury and the role of the pAkt, Nrf2 and hemeoxygenase-1 (HO-1) in the mice brains of permanent middle cerebral artery occlusion (pMCAO).
Related JoVE Video
Community knowledge and experience of mosquitoes and personal prevention and control practices in lhasa, tibet.
Int J Environ Res Public Health
PUBLISHED: 05-05-2014
Show Abstract
Hide Abstract
Since 2009, great public attention has been paid in Lhasa City (Tibet, China) to mosquito bites and accompanying inflammatory complications. However, the potential contribution of knowledge levels, experiences, disease control and preventive practices (KEP) towards mosquitoes has not received much attention. To investigate community KEP concerning mosquitoes in Lhasa, a cross-sectional survey was undertaken in four sub-districts of urban Lhasa in 2012. Questionnaires were designed to collect information regarding socio-demographics and KEP concerning the harmful effects of mosquitoes on participants. The scoring for KEP was developed after consultation of literature. A total of 591 eligible questionnaires were examined. The majority of respondents were female (61.8%) with a mean age of 46 years. Nearly all of the respondents were of Tibetan nationality (97.4%) and living in registered native households (92.7%), who have less than primary school education. The averages of overall score, knowledge score, experience score, and practice score were 9.23, 4.53, 1.80, 2.90, respectively. The registered household with the highest overall score, knowledge score and practice score was non-native. Female subjects with monthly incomes between 1000 and 3000 RMB had higher experience scores. The correlation analysis revealed that significant positive linear correlations existed between knowledge and experience, knowledge and practices, and experience and practices towards mosquitoes. Past experiences with mosquitoes can result in a better knowledge of effective mosquito control practices in the present and the future. Though the average of overall scores related to mosquitoes is high among the participants in Lhasa, however, the knowledge about the ecological habits of mosquitoes should be strengthened. The findings in this study may help to develop strategies and measures of mosquito and mosquito-borne diseases in the future, not only in Lhasa, but also in similar altitude, latitude and longitude regions worldwide.
Related JoVE Video
Plant villins: Versatile actin regulatory proteins.
J Integr Plant Biol
PUBLISHED: 05-04-2014
Show Abstract
Hide Abstract
Regulation of actin dynamics is a central theme in cell biology that is important for different aspects of cell physiology. Villin, a member of the villin/gelsolin/fragmin superfamily of proteins, is an important regulator of actin. Villins contain six gelsolin homology domains (G1-G6) and an extra headpiece domain. In contrast to their mammalian counterparts, plant villins are expressed widely, implying that plant villins play a more general role in regulating actin dynamics. Some plant villins have a defined role in modifying actin dynamics in the pollen tube; most of their in vivo activities remain to be ascertained. Recently, our understanding of the functions and mechanisms of action for plant villins has progressed rapidly; primarily due to the advent of Arabidopsis thaliana genetic approaches and imaging capabilities that can visualize actin dynamics at the single filament level in vitro and in living plant cells. In this review, we focus on discussing the biochemical activities and modes of regulation of plant villins. Here, we present current understanding of the functions of plant villins. Finally, we highlight some of the key unanswered questions regarding the functions and regulation of plant villins for future research.
Related JoVE Video
Homocysteine induces the expression of C-reactive protein via NMDAr-ROS-MAPK-NF-?B signal pathway in rat vascular smooth muscle cells.
Atherosclerosis
PUBLISHED: 05-04-2014
Show Abstract
Hide Abstract
Homocysteine (Hcy) is known as an independent risk factor for atherosclerosis. C-reactive protein (CRP) directly participates in initiation and progression of atherosclerosis. However, there is no direct evidence to demonstrate pro-inflammatory effect of Hcy on vascular smooth muscle cells (VSMCs) through CRP. In the present study, we examined the effect of Hcy on CRP expression and investigated the related mechanism in VSMCs.
Related JoVE Video
Enhanced 4T1 breast carcinoma anticancer activity by co-delivery of doxorubicin and curcumin with core-shell drug-carrier based on heparin modified poly(L-lactide) grafted polyethylenimine cationic nanoparticles.
J Biomed Nanotechnol
PUBLISHED: 04-18-2014
Show Abstract
Hide Abstract
Use of single chemotherapy agents has shown some limitations in anti-tumor treatment, such as development of drug resistance, severe adverse reactions and limited regime for therapeutic use. Combination of two or more therapeutic drugs is a feasible strategy to overcome these limitations. This paper reports study of co-delivery by core-shell nanoparticles (NPs) with hydrophobic PLLA core loaded with curcumin (Cur) and hydrophilic heparin shell adsorbing Doxorubicin (DOX). Characterizations of Cur-PEA NPs, Cur-PEA/heparin NPs and DOX adsorbing into Cur-PEA/heparin NPs (DOX-Cur NPs) were also investigated by transmission electron microscope (TEM) and Malvern Zetasizer. Studies on cellular uptake of DOX-Cur NPs demonstrated that both drugs were effectively taken up by 4T1 tumor cells. Furthermore, DOX-Cur NPs suppressed 4T1 tumor cells growth more efficiently than either DOX or Cur alone at the same concentrations, as measured by flow cytometry (FCM). We found out that intravenous injection of DOX-Cur NPs efficiently inhibited growth of subcutaneous 4T1 breast carcinoma in vivo (p < 0.01) and prolonged survival of the treated 4T1 breast carcinoma mice. Moreover, the pathological damage to the cardiac tissue in mice treated with DOX-Cur NPs was significantly less severe than that of mice treated with free DOX. This study suggested that DOX-Cur NPs may have promising applications in breast carcinoma therapy.
Related JoVE Video
MicroRNA directly enhances mitochondrial translation during muscle differentiation.
Cell
PUBLISHED: 04-15-2014
Show Abstract
Hide Abstract
MicroRNAs are well known to mediate translational repression and mRNA degradation in the cytoplasm. Various microRNAs have also been detected in membrane-compartmentalized organelles, but the functional significance has remained elusive. Here, we report that miR-1, a microRNA specifically induced during myogenesis, efficiently enters the mitochondria where it unexpectedly stimulates, rather than represses, the translation of specific mitochondrial genome-encoded transcripts. We show that this positive effect requires specific miR:mRNA base-pairing and Ago2, but not its functional partner GW182, which is excluded from the mitochondria. We provide evidence for the direct action of Ago2 in mitochondrial translation by crosslinking immunoprecipitation coupled with deep sequencing (CLIP-seq), functional rescue with mitochondria-targeted Ago2, and selective inhibition of the microRNA machinery in the cytoplasm. These findings unveil a positive function of microRNA in mitochondrial translation and suggest a highly coordinated myogenic program via miR-1-mediated translational stimulation in the mitochondria and repression in the cytoplasm.
Related JoVE Video
Microarray of surface-exposed proteins of Rickettsia heilongjiangensis for serodiagnosis of Far-eastern spotted fever.
BMC Infect. Dis.
PUBLISHED: 04-14-2014
Show Abstract
Hide Abstract
Far-eastern spotted fever (FESF) is an important emerging infectious disease in Northeast Asia. The laboratory diagnosis of FESF in hospitals is mainly based on serological methods. However, these methods need to cultivate rickettsial cells as diagnostic antigens, which is both burdensome and dangerous.
Related JoVE Video
Protective effect of SKLB010 against D-galactosamine/lipopolysaccharide-induced acute liver failure via nuclear factor-?B signaling pathway in macrophages.
Int. Immunopharmacol.
PUBLISHED: 04-10-2014
Show Abstract
Hide Abstract
Acute liver failure is characterized by the sudden loss of hepatic function and a high mortality. SKLB010, a derivative of thiazolidinediones, has been proved to be effective in protecting mice from acute liver failure caused by concanavalin A and carbon tetrachloride in our previous work. The purpose of the current study was to evaluate whether SKLB010 could prevent acute liver injury caused by d-galactosamine/lipopolysaccharide (LPS) in mice, and to investigate the underlying mechanisms. In the macrophage-mediated D-GalN/LPS model of acute liver injury, serum enzyme activity was suppressed and liver injury was attenuated by SKLB010. The serum levels of TNF-? and hepatic TNF-? mRNA expression were also markedly decreased after the treatment of SKLB010. In the liver of mice receiving injections of D-GalN/LPS, hepatocytes apoptosis and the infiltration of monocytes/macrophages were blocked by SKLB010. Furthermore, the survival rate of mice following D-GalN/LPS treatment was significantly improved by a single injection with SKLB010. In vivo, the luminescence intensity was suppressed by SKLB010 in NF-?B-luc mice after D-GalN/LPS treatment. In vitro, the production of tumor necrosis factor (TNF)-? and nitrite/nitrate in LPS-stimulated RAW264.7 macrophages was decreased by SKLB010 in a dose-dependent manner. Our further studies demonstrated that SKLB010 inhibited the phosphorylation of I?B? and p38MAPK, and the DNA binding activity of NF-?B in RAW264.7 cells. In conclusion, treatment with only a single injection of SKLB010 could significantly attenuate acute inflammation in mice induced by D-GalN/LPS, and these effects are likely associated with the inhibition of NF-?B activity.
Related JoVE Video
ARHGEF28 gene exon 6/intron 6 junction mutations in Chinese amyotrophic lateral sclerosis cohort.
Amyotroph Lateral Scler Frontotemporal Degener
PUBLISHED: 04-08-2014
Show Abstract
Hide Abstract
It was reported that the intron 6, + 1 del G (GT>TT) mutation of the ARHGEF28 gene generates a shortened protein that might be related to amyotrophic lateral sclerosis (ALS). We sequenced this mutation in 25 familial ALS (FALS), 357 sporadic ALS (SALS) patients, and 442 healthy control subjects. We found just two SALS patients exhibited the mutation so that the incidence of this mutation was 0.52% (2/382) of all the ALS patients. The clinical features of the mutation-positive patients were quite different from the case reported in a previous study. These characteristics differed in terms of gender, site of onset, cognitive function, and family history.
Related JoVE Video
Demethylation of the miR-146a promoter by 5-Aza-2'-deoxycytidine correlates with delayed progression of castration-resistant prostate cancer.
BMC Cancer
PUBLISHED: 04-07-2014
Show Abstract
Hide Abstract
Androgen deprivation therapy is the primary strategy for the treatment of advanced prostate cancer; however, after an initial regression, most patients will inevitably develop a fatal androgen-independent tumor. Therefore, understanding the mechanisms of the transition to androgen independence prostate cancer is critical to identify new ways to treat older patients who are ineligible for conventional chemotherapy.
Related JoVE Video
Development of serous ovarian cancer is associated with the expression of homologous recombination pathway proteins.
Pathol. Oncol. Res.
PUBLISHED: 04-04-2014
Show Abstract
Hide Abstract
To investigate the expressions of key markers in the homologous recombination (HR) pathway and the correlation with clinicopathological parameters in serous ovarian cancer (SOC). We analyzed the protein expression of MRE11, MDC1, ATM, ATR and BRCA1 by immunohistochemistry (IHC) in 97 SOC samples, and correlated with clinical parameters including age, tumor grades, clinical stage, status of menstruation and chemotherapy. Low expression of MRE11 and MDC1 was detected in 14.4 % and 3.1 % of the patient samples, and negative expression of ATM, ATR and BRCA1 was found in 11.3 %, 6.3 % and 29.9 % of the patient samples, respectively. ATR deficiency was significantly associated with menopause (P?=?0.025), strong expression of ATM (P?=?0.017) and MRE11 (P?=?0.040) with pre-menopausal SOC, strong expression of MRE11 (P?=?0.016) with low tumor grade, and strong expression of BRCA1 (P?=?0.015) with early clinical stage. In addition, low expression of MRE11 was strongly associated with negativity of ATR (P?
Related JoVE Video
Palmitic acid exerts pro?inflammatory effects on vascular smooth muscle cells by inducing the expression of C?reactive protein, inducible nitric oxide synthase and tumor necrosis factor??.
Int. J. Mol. Med.
PUBLISHED: 03-28-2014
Show Abstract
Hide Abstract
Atherosclerosis is a chronic inflammatory disease in the vessel, and inflammatory cytokines play an important role in the inflammatory process of atherosclerosis. A high level of free fatty acids (FFAs) produced in lipid metabolism disorders are known to participate in the formation of atherosclerosis through multiple bioactivities. As the main saturated fatty acid in FFAs, palmitic acid stimulates the expression of inflammatory cytokines in macrophages. However, it is unclear whether palmitic acid exerts a pro?inflammatory effect on vascular smooth muscle cells (VSMCs). The purpose of the present study was to observe the effect of palmitic acid on the expression of C?reactive protein (CRP), tumor necrosis factor ? (TNF??) and inducible nitric oxide synthase (iNOS) in VSMCs. Rat VSMCs were cultured, and palmitic acid was used as a stimulant for CRP, TNF?? and iNOS expression. mRNA expression was assayed with reverse transcription?polymerase chain reaction, and protein expression was detected with western blot analysis and immunocytochemistry. The results showed that palmitic acid significantly stimulated mRNA and protein expression of CRP, TNF?? and iNOS in VSMCs in time? and concentration?dependent manners, and therefore, palmitic acid is able to exert a pro?in?ammatory effect on VSMCs via stimulating CRP, TNF?? and iNOS expression. The ?ndings provide a novel explanation for the direct pro?in?ammatory and atherogenic effects of palmitic acid, and for the association with metabolic syndrome, such as type 2 diabetes mellitus, obesity and atherosclerosis. Therefore, the intervention with anti?inflammatory agents may effectively delay the formation and progression of atherosclerosis in patients with metabolic syndrome.
Related JoVE Video
Insulin increases phosphorylation of mitochondrial proteins in human skeletal muscle in vivo.
J. Proteome Res.
PUBLISHED: 03-28-2014
Show Abstract
Hide Abstract
There is increasing evidence that multiple proteins involved in key regulatory processes in mitochondria are phosphorylated in mammalian tissues. Insulin regulates glucose metabolism by phosphorylation-dependent signaling and has been shown to stimulate ATP synthesis in human skeletal muscle. Here, we investigated the effect of insulin on the phosphorylation of mitochondrial proteins in human skeletal muscle in vivo. Using a combination of TiO(2) phosphopeptide-enrichment, HILIC fractionation, and LC-MS/MS, we compared the phosphoproteomes of isolated mitochondria from skeletal muscle samples obtained from healthy individuals before and after 4 h of insulin infusion. In total, we identified 207 phosphorylation sites in 95 mitochondrial proteins. Of these phosphorylation sites, 45% were identified in both basal and insulin-stimulated samples. Insulin caused a 2-fold increase in the number of different mitochondrial phosphopeptides (87 ± 7 vs 40 ± 7, p = 0.015) and phosphoproteins (46 ± 2 vs 26 ± 3, p = 0.005) identified in each mitochondrial preparation. Almost half of the mitochondrial phosphorylation sites (n = 94) were exclusively identified in the insulin-stimulated state and included the majority of novel sites. Phosphorylation sites detected more often or exclusively in insulin-stimulated samples include multiple sites in mitochondrial proteins involved in oxidative phosphorylation, tricarboxylic acid cycle, and fatty acid metabolism, as well as several components of the newly defined mitochondrial inner membrane organizing system (MINOS). In conclusion, the present study demonstrates that insulin increases the phosphorylation of several mitochondrial proteins in human skeletal muscle in vivo and provides a first step in the understanding of how insulin potentially regulates mitochondrial processes by phosphorylation-dependent mechanisms.
Related JoVE Video
Comparative proteomic analysis of histone post-translational modifications upon ischemia/reperfusion-induced retinal injury.
J. Proteome Res.
PUBLISHED: 03-26-2014
Show Abstract
Hide Abstract
We present a detailed quantitative map of single and coexisting histone post-translational modifications (PTMs) in rat retinas affected by ischemia and reperfusion (I/R) injury. Retinal I/R injury contributes to serious ocular diseases, which can lead to vision loss and blindness. We applied linear ion trap-orbitrap hybrid tandem mass spectrometry (MS/MS) to quantify 131 single histone marks and 143 combinations of multiple histone marks in noninjured and injured retinas. We observed 34 histone PTMs that exhibited significantly (p < 0.05) different abundance between healthy and I/R injured eyes, of which we confirmed three H4 histone marks by Western blotting. H4K20me2 was up to 4-fold change up-regulated after the injury and is associated with the response to DNA damage as demonstrated by an increase in the phosphorylation of p53 and Chk1. This study demonstrates that quantitative MS provides a sensitive and accurate way to dissect the changes in the histone code after retinal injury. Specifically, DNA damage associated histone PTMs may contribute to neurovascular degeneration during the process of ischemia/reperfusion injury.
Related JoVE Video
Vapor-phase hydrothermal synthesis of rutile TiO? nanostructured film with exposed pyramid-shaped (111) surface and superiorly photoelectrocatalytic performance.
J Colloid Interface Sci
PUBLISHED: 03-25-2014
Show Abstract
Hide Abstract
Rutile TiO2 nanostructured film with exposed pyramid-shaped (111) surface was successfully fabricated using metal titanium foil as substrate through a facile vapor-phase hydrothermal method. The fabricated rutile TiO2 film was composed of vertically aligned rod-like structures with diameters ranged from 400 to 700 nm and thickness of ca. 2.0 ?m. The obtained rutile TiO2 film as photoanode exhibited excellent photoelectrocatalytic activity toward water oxidation and rhodamine B decolorization under UV illumination, which was more than 3.5 and 1.2 times of that obtained by highly ordered anatase TiO2 nanotube array film photoanode under the same experimental conditions, respectively. The excellent photoelectrocatalytic performance of the rutile TiO2 film photoanode could be due to the superior photoelectron transfer property and the high oxidative capability of {111} crystal facets. The superior photoelectron transfer capability of the photoanodes was manifested by the inherent resistance (R0) of the photoanodes using a simple photoelectrochemical method. The calculated R0 values were 50.5 and 86.2 ? for the rutile TiO2 nanostructured film and anatase TiO2 nanotube array film, respectively. Lower R0 value of the rutile TiO2 photoanode indicated a superior photoelectron transfer capability owing to good single crystal property of the rod-like rutile nanostructure. Almost identical valence band level (1.94 eV) of the rutile TiO2 nanostructured film and anatase TiO2 nanotube array film (meaning a similar oxidation capability) further confirmed the significant role of photoelectron transfer capability and exposed high-energy {111} crystal facets for improved photoelectrocatalytic performance of the rutile TiO2 nanostructured film photoanode.
Related JoVE Video
A new electrochemical aptasensor for the analysis of the vascular endothelial growth factor.
J Immunoassay Immunochem
PUBLISHED: 03-25-2014
Show Abstract
Hide Abstract
The vascular endothelial growth factor (VEGF) is a mitogen and important regulator of angiogenesis by activation the VEGF-receptor tyrosine kinases in endothelial cells. VEGF finds growing interest as a therapeutic target, and its analytical detection has important implications as biomarker for different clinical disorders. Herein, we propose a new strategy to fabricate elegant electrochemical aptasensors, and this strategy has been employed to develop a biosensor for the assay of the concentration of VEGF. In our strategy, an electroactive probe, hemin, is not covalently modified at the end of the oligonucleotide. The method was simple and economical, allowing monitoring of VEGF to low concentrations of 1 nM.
Related JoVE Video
Abnormal expression of EMT-related proteins, S100A4, vimentin and E-cadherin, is correlated with clinicopathological features and prognosis in HCC.
Med. Oncol.
PUBLISHED: 03-24-2014
Show Abstract
Hide Abstract
We determined the expression of epithelial-mesenchymal transition (EMT) indicator proteins, E-cadherin (E-cad), vimentin (VIM), mucin 1 (MUC1) and S100 calcium-binding protein A4 (S100A4) in hepatocellular carcinoma (HCC) patient tissue samples. We also investigated the relationship between the expression of these proteins and clinicopathologic factors in HCC. Finally, we assessed the potential value of these markers as prognostic indicators of survival in HCC patients. The expression of E-cad, VIM, MUC1 and S100A4 EMT indicator proteins was assessed in tissue microarray HCC tissue sections and corresponding peritumoral normal tissues by immunohistochemistry. In addition, the expression for the four EMT indicator proteins was correlated with clinicopathological features of HCC and patient outcome. Comparison of clinicopathological characteristics and immunohistochemistry by ?(2) analysis revealed that downregulation of E-cad in HCC was significantly associated with later TNM cancer stage (P = 0.012), gross classification (P = 0.018), regional lymph node metastasis (P = 0.036) and liver cirrhosis (P = 0.028). Increased S100A4 expression in HCC was significantly associated with differentiation (P = 0.032), tumor with a complete fibrous capsule (P = 0.031) and portal vein invasion (P = 0.038). High VIM expression in HCC was significantly associated with high serum ?-fetoprotein levels (P = 0.016). We also observed that low E-cad expression was significantly associated with overexpression of VIM (P = 0.001). Kaplan-Meier survival and Cox regression analysis revealed that low E-cad expression (HR = 0.164, 95 % CI 0.072 to 0.373, P < 0.001) and high serum ?-fetoprotein levels (HR = 2.202, 95 % CI 1.054 to 4.598, P = 0.036) were independent prognostic factors in HCC. Our study demonstrates that high S100A4 and VIM expression and low E-cad expression correlate with an aggressive, malignant phenotype in HCC. These results also support a role for E-cad as a prognostic factor in HCC.
Related JoVE Video
Hydrothermal transformation of dried grass into graphitic carbon-based high performance electrocatalyst for oxygen reduction reaction.
Small
PUBLISHED: 03-23-2014
Show Abstract
Hide Abstract
In this work, we present a low cost and environmentally benign hydrothermal method using dried grass as the sole starting material without any synthetic chemicals to directly produce high quality nitrogen-doped carbon nanodot/nanosheet aggregates (N-CNAs), achieving a high yield of 25.2%. The fabricated N-CNAs possess an N/C atomic ratio of 3.41%, consist of three typed of doped N at a ratio of 2.6 (pyridinic):1.7 (pyrrolic):1 (graphitic). The experimental results reveal that for oxygen reduction reaction (ORR), the performance of N-CNAs, in terms of electrocatalytic activity, stability and resistance to crossover effects, is better or comparable to the commercial Pt/C electrocatalyst. The theoretical studies further indicate that the doped pyridinic-N plays a key role for N-CNAs' excellent four-electron ORR electrocatalytic activity.
Related JoVE Video
Stabilized human TRIM5? protects human T cells from HIV-1 infection.
Mol. Ther.
PUBLISHED: 03-19-2014
Show Abstract
Hide Abstract
Rhesus (rh) but not human (hu) TRIM5? potently restricts human immunodeficiency virus (HIV)-1 infection. It is not clear why huTRIM5? fails to effectively block HIV infection, but it is thought to have a lower affinity for the viral core. Using primary human CD4 T cells, we investigated the ability of huTRIM5?, rhTRIM5?, and the huTRIM5?R323-332 B30.2/SPRY patch-mutant to form cytoplasmic bodies, postulated as key components of the HIV-1 restriction apparatus. Both rhTRIM5? and huTRIM5?R323-332 formed pronounced cytoplasmic bodies, whereas cytoplasmic bodies in T cells overexpressing huTRIM5? were present but more difficult to detect. As expression of all three TRIM5? orthologs was similar at the RNA level, we next investigated the role of protein stability in conferring TRIM5?-mediated HIV-1 restriction. Both steady-state and pulse-chase experiments revealed that the huTRIM5? protein was much less stable than rhTRIM5?, and this difference correlated with higher self-ubiquitination activity. Using a stabilized form of huTRIM5? in which the steady-state expression level was more similar to rhTRIM5?, we observed comparable HIV-1 restriction activity in multi-round HIV-1 challenge assays. Lastly, primary human CD4 T cells expressing a stabilized huTRIM5? were protected from HIV-1-mediated destruction in vivo, indicating that efforts to stabilize huTRIM5? should have significant long-term therapeutic value.
Related JoVE Video
The oncogenic role of PKCiota gene amplification and overexpression in Chinese non-small cell lung cancer.
Lung Cancer
PUBLISHED: 03-19-2014
Show Abstract
Hide Abstract
The atypical protein kinase C isozyme iota (PKCiota) has been proposed as an oncogene based on its transformation property and amplification identified in Caucasian non-small cell lung cancer (NSCLC) patients. Because the geography difference of some genetic aberrance such as EGFR mutations between Caucasian and Asian NSCLC patients has been identified previously, it is important to know whether the PKCiota amplification also occurs in Asian NSCLC patients.
Related JoVE Video
Chrysophanol inhibits NALP3 inflammasome activation and ameliorates cerebral ischemia/reperfusion in mice.
Mediators Inflamm.
PUBLISHED: 03-15-2014
Show Abstract
Hide Abstract
The most effective way to contain cerebral ischemic injury is reperfusion; however, reperfusion itself may result in tissue injury, for which inflammatory damage is one of the main causative factors. NALP3 inflammasome is a multiprotein complex. It consists of NALP3, ASC, and caspase-1, whose function is to switch on the inflammatory process. Chrysophanol is an extract from plants of Rheum genus and it possesses many pharmacological effects including its anti-inflammation activity. In this study, the effects of chrysophanol in cerebral ischemia/reperfusion and the potential mechanisms were investigated. Male CD1 mice were subject to transient middle cerebral artery occlusion (tMCAO). The NALP3 inflammasome activation status and its dynamic expression during the natural inflammatory response induced by tMCAO were first profiled. The neuroprotective effects of chrysophanol were then assessed and the potential mechanisms mediating the observed neuroprotection were then explored. Physical parameters including neurological deficit, infarct size, brain edema, and BBB permeability were measured at 24 h after tMCAO. Confocal microscopy, Western blotting, immunohistochemistry, and qRT-PCR techniques were utilized to analyze the expression of NALP3 inflammasome and IL-1 ? . Our results indicated that the brain tissue damage during cerebral ischemia/reperfusion is accompanied by NALP3 inflammasome activation. Chrysophanol could inhibit the activation of NALP3 inflammasome and protect cerebral ischemic stroke.
Related JoVE Video
Genomic and comparative genomic analyses of Rickettsia heilongjiangensis provide insight into its evolution and pathogenesis.
Infect. Genet. Evol.
PUBLISHED: 03-05-2014
Show Abstract
Hide Abstract
Rickettsia heilongjiangensis, the causative agent of far eastern spotted fever, is an obligate intracellular gram-negative bacterium that belongs to the spotted fever group rickettsiae. To understand the evolution and pathogenesis of R. heilongjiangensis, we analyzed its genome and compared it with other rickettsial genomes available in GenBank. The R. heilongjiangensis chromosome contains 1333 genes, including 1297 protein coding genes and 36 RNA coding genes. The genome also contains 121 pseudogenes, 54 insertion sequences, and 39 tandem repeats. Sixteen genes encoding the major components of the type IV secretion systems were identified in the R. heilongjiangensis genome. In total, 37 ?-barrel outer membrane proteins were predicted in the genome, eight of which have been previously confirmed to be outer membrane proteins. In addition, 266 potential virulence factor genes, seven partially deleted antibiotic resistance genes, and a genomic island were identified in the genome. The codon usage in the genome is compatible with its low GC content, and the amino acid usage shows apparent bias. A comparative genomic analysis showed that R. heilongjiangensis and R. japonica share one unique fragment that may be a target sequence for a diagnostic assay. The orthologs of 37 genes of R. heilongjiangensis were found in pathogenic R. rickettsii str. Sheila Smith but not in non-pathogenic R. rickettsii str. Iowa, which may explain why R. heilongjiangensis is pathogenic. Pan-genome analysis showed that R. heilongjiangensis and 42 other rickettsiae strains share 693 core genes with a pan-genome size of 4837 genes. The pan-genome-based phylogeny showed that R. heilongjiangensis was closely related to R. japonica.
Related JoVE Video
Protective immunity against Rickettsia heilongjiangensis in a C3H/HeN mouse model mediated by outer membrane protein B-pulsed dendritic cells.
Sci China Life Sci
PUBLISHED: 03-02-2014
Show Abstract
Hide Abstract
Rickettsia heilongjiangensis is an obligate intracellular bacterium that causes Far-Eastern tick-borne spotted fever. Outer membrane protein B (OmpB) is an important surface protein antigen of rickettsiae. In the present study, the ompB gene of R. heilongjiangensis was divided into four fragments, resulting in four recombinant proteins (OmpB-p1, OmpB-p2, OmpB-p3, and OmpB-p4). Each OmpB was used in vitro to stimulate murine bone marrow-derived dendritic cells (BMDCs) of C3H/HeN mice, and the OmpB-pulsed BMDCs were transferred to naïve C3H/HeN mice. On day 14 post-transfer of BMDCs, the mice were challenged with R. heilongjiangensis and the rickettsial loads in the mice were quantitatively determined on day 7 post-challenge. Mice receiving BMDCs pulsed with OmpB-p2, OmpB-p3, or OmpB-p4 exhibited significantly lower bacterial load compared with mice receiving OmpB-p1-pulsed BMDCs. CD4(+) and CD8(+) T cells isolated from the spleen of C3H/HeN mice receiving BMDCs pulsed with each OmpB were co-cultured with BMDCs pulsed with the respective cognate protein. In flow cytometric analysis, the expression level of CD69 on CD4(+) or CD8(+) T cells from mice receiving BMDCs pulsed with OmpB-p2, OmpB-p3, or OmpB-p4 was higher than that on cells from mice receiving OmpB-p1-pulsed BMDCs, while the expression level of tumor necrosis factor (TNF)-? on CD8(+) T cells and interferon (IFN)-? on the CD4(+) and CD8(+) T cells from mice receiving OmpB-p2, -p3, or -p4 was significantly higher than on cells from mice receiving OmpB-p1-pulsed BMDCs. Our results suggest that the protective OmpBs could activate CD4(+) and CD8(+) T cells and drive their differentiation toward CD4(+) Th1 and CD8(+) Tcl cells, respectively, which produce greater amounts of TNF-? and, in particular, IFN-?, to enhance rickettsicidal activity of host cells.
Related JoVE Video
Quantitative proteomics reveals olfactory input-dependent alterations in the mouse olfactory bulb proteome.
J Proteomics
PUBLISHED: 03-01-2014
Show Abstract
Hide Abstract
Olfactory sensory information is processed and integrated by circuits within the olfactory bulb (OB) before being sent to the olfactory cortex. In the mammalian OB, neural activity driven by external stimuli can lead to experience-dependent changes in structures and functions. In this study, quantitative proteomics techniques were employed to study proteome-wide changes in the OB under four levels of neural activity (from low to high): devoid of peripheral input (using a transgenic model), wild-type control, and short-term and long-term odor exposures. Our results revealed that proteins related to various processes were altered in the OBs of odor-deprived and odor-stimulated mice compared to the wild-type controls. These changes induced by odor stimulation were quite different from those induced by a deficit in peripheral olfactory inputs. Detailed analysis demonstrated that metabolic process and synaptic transmission were the most commonly altered pathways and that the effects of peripheral deprivation were more profound. Our comparative proteomics analysis indicated that olfactory deprivation and odor exposure lead to different alterations in the OB proteome, which provides new clues about the mechanisms underlying the olfactory deprivation- or odor stimulation-induced plasticity of OB function and organization.
Related JoVE Video
100 Gy 60 Co ?-ray induced novel mutations in tetraploid wheat.
ScientificWorldJournal
PUBLISHED: 02-27-2014
Show Abstract
Hide Abstract
10 accessions of tetraploid wheat were radiated with 100 Gy (60)Co ? -ray. The germination energy, germination rate, special characters (secondary tillering, stalk with wax powder, and dwarf), meiotic process, and high-molecular-weight glutenin subunits (HMW-GSs) were observed. Different species has different radiation sensibility. With 1 seed germinated (5%), T. dicoccum (PI434999) is the most sensitive to this dose of radiation. With a seed germination rate of 35% and 40%, this dose also affected T. polonicum (As304) and T. carthlicum (As293). Two mutant dwarf plants, T. turgidum (As2255) 253-10 and T. polonicum (As302) 224-14, were detected. Abnormal chromosome pairings were observed in pollen mother cells of both T. dicoccoides (As835) 237-9 and T. dicoccoides (As838) 239-8 with HMW-GS 1Ax silent in seeds from them. Compared with the unirradiated seed of T. polonicum (As304) CK, a novel HMW-GS was detected in seed of T. polonicum (As304) 230-7 and its electrophoretic mobility was between 1By8 and 1Dy12 which were the HMW-GSs of Chinese Spring. These mutant materials would be resources for wheat breeding.
Related JoVE Video
Association of sequence polymorphism in the mitochondrial D-loop with chronic kidney disease.
Ren Fail
PUBLISHED: 02-27-2014
Show Abstract
Hide Abstract
The mitochondrial displacement loop (D-loop) is known to accumulate mutations and single nucleotide polymorphisms (SNPs) at a higher frequency than other regions of mitochondrial DNA (mtDNA).
Related JoVE Video
MicroRNA-93 inhibits inflammatory cytokine production in LPS-stimulated murine macrophages by targeting IRAK4.
FEBS Lett.
PUBLISHED: 02-23-2014
Show Abstract
Hide Abstract
Endotoxin-induced uveitis (EIU) is an animal model of acute ocular inflammation for the study of human endogenous anterior uveitis. The mechanisms accounting for the development of ocular inflammation remain hazy. MicroRNAs (mi-RNAs) have been proposed as novel regulators of inflammation. It remains unclear whether a microRNA-mediated regulatory mechanism is involved in LPS-induced EIU. In this study, we report that miR-93 expression in the eyes of EIU rats and LPS-stimulated macrophages is significantly decreased. We also show that miR-93 inhibits NF-?B activation and pro-inflammatory cytokines by targeting IRAK4 expression. We further demonstrate that miR-93 inhibits IRAK4 expression by binding directly to the 3'-UTR of IRAK4. Our findings suggest that miR-93 is a negative regulator of the immune response in EIU.
Related JoVE Video
Discovery and optimization of indazoles as potent and selective interleukin-2 inducible T cell kinase (ITK) inhibitors.
Bioorg. Med. Chem. Lett.
PUBLISHED: 02-20-2014
Show Abstract
Hide Abstract
There is evidence that small molecule inhibitors of the non-receptor tyrosine kinase ITK, a component of the T-cell receptor signaling cascade, could represent a novel asthma therapeutic class. Moreover, given the expected chronic dosing regimen of any asthma treatment, highly selective as well as potent inhibitors would be strongly preferred in any potential therapeutic. Here we report hit-to-lead optimization of a series of indazoles that demonstrate sub-nanomolar inhibitory potency against ITK with strong cellular activity and good kinase selectivity. We also elucidate the binding mode of these inhibitors by solving the X-ray crystal structures of the complexes.
Related JoVE Video
Surface protein Adr2 of Rickettsia rickettsii induced protective immunity against Rocky Mountain spotted fever in C3H/HeN mice.
Vaccine
PUBLISHED: 02-10-2014
Show Abstract
Hide Abstract
Rickettsia rickettsii is the pathogen of Rocky Mountain spotted fever (RMSF), a life-threatening tick-transmitted infection. Adr2 was a surface-exposed adhesion protein of R. rickettsii and its immunoprotection against RMSF was investigated in mice.
Related JoVE Video
A cellular system that degrades misfolded proteins and protects against neurodegeneration.
Mol. Cell
PUBLISHED: 01-31-2014
Show Abstract
Hide Abstract
Misfolded proteins compromise cellular function and cause disease. How these proteins are detected and degraded is not well understood. Here we show that PML/TRIM19 and the SUMO-dependent ubiquitin ligase RNF4 act together to promote the degradation of misfolded proteins in the mammalian cell nucleus. PML selectively interacts with misfolded proteins through distinct substrate recognition sites and conjugates these proteins with the small ubiquitin-like modifiers (SUMOs) through its SUMO ligase activity. SUMOylated misfolded proteins are then recognized and ubiquitinated by RNF4 and are subsequently targeted for proteasomal degradation. We further show that PML deficiency exacerbates polyglutamine (polyQ) disease in a mouse model of spinocerebellar ataxia 1 (SCA1). These findings reveal a mammalian system that removes misfolded proteins through sequential SUMOylation and ubiquitination and define its role in protection against protein-misfolding diseases.
Related JoVE Video
Distinctive effects of the cellular inhibitor of apoptosis protein c-IAP2 through stabilization by XIAP in glioblastoma multiforme cells.
Cell Cycle
PUBLISHED: 01-22-2014
Show Abstract
Hide Abstract
Inhibitor of apoptosis proteins (IAPs) are extensively involved in NF?B signaling pathways. Regulation of c-IAP2 turnover by other proteins was investigated in glioblastoma multiforme (GBM) cells in the present study. When overexpressed, X-linked IAP (XIAP) enhanced expression of ectopic c-IAP2, but not c-IAP1, and endogenous c-IAP2 levels were reduced once XIAP expression was silenced. TNF? stimulation substantially increased c-IAP2 expression, and this upregulation was impaired by suppression of XIAP. Similarly, when XIAP was limiting due to severe hypoxic conditions, c-IAP2 levels were downregulated. These data together indicate that XIAP is an important regulator responsible for stabilization of c-IAP2 levels under different conditions. Protein interactions occur through binding of BIR2 and BIR3 domains of c-IAP2 with the RING finger of XIAP. XIAP inhibition of c-IAP2 auto-degradation was dependent on this physical interaction, and it was independent of XIAP E3 ligase activity. Global c-IAP2 ubiquitination was not affected by XIAP, although c-IAP2 levels were significantly increased. A CARD-RING-containing fragment of c-IAP2 was found to target XIAP for proteasome-independent degradation, but it was unable to sensitize GBM cells to chemo-reagents. The XIAP-stabilized c-IAP2 was found to enhance I?B-? phosphorylation on serines 32 and 36, and to antagonize XIAP-induced increase in mature Smac and Bcl10. Taken together, our data identify a distinctive role of c-IAP2 as stabilizer of XIAP, which is likely involved in regulation of NF?B activation and apoptosis in GBM cells.
Related JoVE Video
Ellagic acid-induced thrombotic focal cerebral ischemic model in rats.
J Pharmacol Toxicol Methods
PUBLISHED: 01-02-2014
Show Abstract
Hide Abstract
Ischemic stroke is a common cause of human disability and death. Animal models of focal cerebral ischemia are widely utilized to mimic human ischemic stroke. Although models of focal cerebral ischemia have been well established, very few evidence is based on triggering the intrinsic coagulation system to induce focal cerebral ischemia. Ellagic acid (EA) has been identified to trigger the intrinsic coagulation system via activating coagulation factor XII. However, it remains unknown whether EA can serve as a novel pharmacological approach to induce a new model of focal cerebral ischemia in rats.
Related JoVE Video
Upregulation of Atoh1 correlates with favorable survival in gastrointestinal stromal tumor.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Atonal homolog 1 (Atoh1) is crucial to the differentiation of many cell types and participates in tumorigenesis and progression. However, the expression of Atoh1 in gastrointestinal stromal tumors (GIST) and its relationship to clinical characteristics of this disease remain poorly understood. In this study, immunohistochemical analysis using tissue microarray (TMA) was employed to evaluate the expression of Atoh1 in GIST and the correlation between Atoh1 expression and clinicopathological features of GIST as well as patient outcome. High Atoh1 cytoplasmic expression was observed in 77.22% of patients with GIST, which was related to the mitotic index (P = 0.010) and AFIP-Miettinen risk classification (P = 0.045). High Atoh1 nuclear expression was seen in 69.49% of cases, which was associated with mitotic index (P = 0.003) and AFIP-Miettinen risk classification (P = 0.001). The Kaplan-Meier method and log-rank test indicated that high Atoh1 cytoplasmic expression, high Atoh1 nuclear expression, small tumor diameter, low mitotic index and TNM stage significantly correlated with improved survival of GIST patients. Overall, the data suggest that Atoh1 high expression correlates with a good prognosis and it may serve as a favorable prognostic factor for GIST. These results also support a role for Atoh1 as a tumor suppressor gene in GIST.
Related JoVE Video
Effect of cyanotoxins on the hypothalamic-pituitary-gonadal axis in male adult mouse.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Microcystins LR (MC-LR) are hepatotoxic cyanotoxins that have been shown to induce reproductive toxicity, and Hypothalamic-Pituitary-Gonadal Axis (HPG) is responsible for the control of reproductive functions. However, few studies have been performed to evaluate the effects of MC-LR on HPG axis. This study aimed to investigate the MC-LR-induced toxicity in the reproductive system of mouse and focus on the HPG axis.
Related JoVE Video
An updated meta-analysis of the association between tumor necrosis factor-? -308G/A polymorphism and obstructive sleep apnea-hypopnea syndrome.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Several studies have reported that the tumor necrosis factor-? (TNF-?) -308G/A polymorphism is associated with susceptibility to obstructive sleep apnea-hypopnea syndrome (OSAHS). However, these results are controversial and conflicting.
Related JoVE Video
Identification of novel surface-exposed proteins of Rickettsia rickettsii by affinity purification and proteomics.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever, is the most pathogenic member among Rickettsia spp. Surface-exposed proteins (SEPs) of R. rickettsii may play important roles in its pathogenesis or immunity. In this study, R. rickettsii organisms were surface-labeled with sulfo-NHS-SS-biotin and the labeled proteins were affinity-purified with streptavidin. The isolated proteins were separated by two-dimensional electrophoresis, and 10 proteins were identified among 23 protein spots by electrospray ionization tandem mass spectrometry. Five (OmpA, OmpB, GroEL, GroES, and a DNA-binding protein) of the 10 proteins were previously characterized as surface proteins of R. rickettsii. Another 5 proteins (Adr1, Adr2, OmpW, Porin_4, and TolC) were first recognized as SEPs of R. rickettsii herein. The genes encoding the 5 novel SEPs were expressed in Escherichia coli cells, resulting in 5 recombinant SEPs (rSEPs), which were used to immunize mice. After challenge with viable R. rickettsii cells, the rickettsial load in the spleen, liver, or lung of mice immunized with rAdr2 and in the lungs of mice immunized with other rSEPs excluding rTolC was significantly lower than in mice that were mock-immunized with PBS. The in vitro neutralization test revealed that sera from mice immunized with rAdr1, rAdr2, or rOmpW reduced R. rickettsii adherence to and invasion of vascular endothelial cells. The immuno-electron microscopic assay clearly showed that the novel SEPs were located in the outer and/or inner membrane of R. rickettsii. Altogether, the 5 novel SEPs identified herein might be involved in the interaction of R. rickettsii with vascular endothelial cells, and all of them except TolC were protective antigens.
Related JoVE Video
simple hit counter

What is Visualize?

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

How does it work?

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

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

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