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.
Analysis of Trace Microcystins in Vegetables Using Solid-phase Extraction Followed by High Performance Liquid Chromatography Triple-quadrupole Mass Spectrometry.
J. Agric. Food Chem.
PUBLISHED: 11-14-2014
Show Abstract
Hide Abstract
A selective and sensitive method for the simultaneous detection of three common and hazardous microcystins-LR,-RR and -YR in various vegetables was established using solid phase extraction followed by high-performance liquid chromatography coupled with mass spectrometry. The methanol-water proportion ratio of the extraction solvent and its acidity, as well as the efficiencies of solid-phase extraction were evaluated to optimize a pre-treatment procedure for extracting the microcystins from 10 vegetable matrices. The limits of detection and quantitation were below 7.5 µg/kg (dw) and 25 µg/kg (dw) respectively in different vegetable matrices. The recoveries of the microcystins in the 10 vegetable matrices ranged from 61.3 to 117.3%, with RSDs of 0.2-18.3%. The established method was used to analyze 28 field vegetable samples collected from the sides of Lake Dianchi, and microcystin-RR was found in almost all samples at concentrations of 36.4-2352.2 µg/kg (dw).
Related JoVE Video
[Adaptive EEMD residue related baseline correction algorithm].
Guang Pu Xue Yu Guang Pu Fen Xi
PUBLISHED: 11-01-2014
Show Abstract
Hide Abstract
Baseline correction is an important part of spectral analysis; the existing algorithms usually need to set the key parameters and does not have adaptability. The spectral baseline is fitted by the residue according to the feature of ensemble empirical mode decomposition (EEMD for short). The correlation between residual and original signal, the self-correlation and the cross-correlation of residual form the residual related rule. The residual related rule is proposed to judge whether the residual is a component of baseline, based on which adaptive EEMD residual related base line algorithm is proposed. With experiment on the simulated spectrum data of superimposing curve background and the linear background, the results showed that in the case of known baseline mathematical assumption: EEMD residual related method is not so good for polynomial fitting, it is almost no difference from linear fitting, but is better than the wavelet decomposition. In the absence of spectral background knowledge, the real Raman spectrum data are tested. The model is established between Raman spectra treated by the procedure above and chlorophyll, and the model corrected by EEMD residual related baseline method has the biggest correlation coefficient and prediction coefficient, but the smallest root mean square error of cross validation and relative prediction deviation. The effect of EEMD residual related baseline method effects on the peak position, peak intensity and peak width is the smallest in all kinds of baseline correction methods. EEMD residual method has the best baseline correction effect. Experiments show that this algorithm can be used for Raman spectra baseline correction, without prior knowledge of the sample composition analysis, and there is no need to select appropriate fitting function, fitting data points, fitting order as well as basis function and decomposition levels, also there is no need of mathematical hypothesis of baseline signal distribution, so the adaptability is very strong.
Related JoVE Video
The relationship of kinase insert domain receptor gene polymorphisms and clinical outcome in advanced hepatocellular carcinoma patients treated with sorafenib.
Med. Oncol.
PUBLISHED: 09-03-2014
Show Abstract
Hide Abstract
Kinase insert domain receptor (KDR) is the principal receptor that promotes the pro-angiogenic action of vascular endothelial growth factor and has been the principal target of anti-angiogenic therapies. Our aim was to determine whether single-nucleotide polymorphisms (SNPs) in KDR gene are associated with clinical outcomes after first-line sorafenib therapy in advanced hepatocellular carcinoma (HCC). The SNPs in KDR were tested in 78 advanced HCC patients receiving first-line sorafenib. Correlations with clinicopathological features and survival were analyzed. Patients with AA genotype of rs1870377 and AA genotype of rs2305948 were significantly associated with a better response and longer time to progression (TTP) (5.8 vs 4.0 months, P=0.001; 5.8 vs 4.5 months, P=0.016, respectively). Patients harboring AA genotype in rs1870377 and TT/TC genotype in rs2071559 had a longer overall survival (OS) (15.0 vs 9.6 months, P=0.001; 13.0 vs 9.0 months, P=0.007, respectively). At multivariate analysis, major vascular invasion and rs1870377 were independent factors in TTP and performance status, rs1870377, and rs2071559 were independent factors in OS. Our results suggest that SNPs in KDR gene can predict clinical outcome in advanced HCC patients receiving first-line sorafenib.
Related JoVE Video
Cyclin D1b splice variant promotes ?v?3-mediated adhesion and invasive migration of breast cancer cells.
Cancer Lett.
PUBLISHED: 09-01-2014
Show Abstract
Hide Abstract
Cyclin D1b, a splice variant of the cell cycle regulator cyclin D1, holds oncogenic functions in human cancer. However, the mechanisms underlying cyclin D1b function remain poorly understood. Here we introduced wild-type cyclin D1a or cyclin D1b variant into non-metastatic MCF-7 cells. Our results show that ectopic expression of cyclin D1b promotes invasiveness of the cancer cells in a cyclin D1a independent manner. Specifically, cyclin D1b is found to modulate the expression of ?v?3, which characterizes the metastatic phenotype, and enhance tumor cell invasive potential in cooperating with HoxD3. Notably, cyclin D1b promotes ?v?3-mediated adhesion and invasive migration, which are associated with invasive potential of breast cancer cells. Further exploration indicates that cyclin D1b makes breast cancer cells more sensitive to toll-like receptor 4 ligand released from damaged tumor cells. These findings reveal a role of cyclin D1b as a possible mediator of ?v?3 transcription to promote tumor metastasis.
Related JoVE Video
Shortest path analyses in the protein-protein interaction network of NGAL (neutrophil gelatinase-associated lipocalin) overexpression in esophageal squamous cell carcinoma.
Asian Pac. J. Cancer Prev.
PUBLISHED: 08-30-2014
Show Abstract
Hide Abstract
NGAL (neutrophil gelatinase-associated lipocalin) is a novel cancer-related protein involves multiple functions in many cancers and other diseases. We previously overexpressed NGAL to analyze its role in esophageal squamous cell carcinoma (ESCC). In this study, a protein-protein interaction (PPI) was constructed and the shortest paths from NGAL to transcription factors in the network were analyzed. We found 28 shortest paths from NGAL to RELA, most of them obeying the principle of extracellular to cytoplasm, then nucleus. These shortest paths were also prioritized according to their normalized intensity from the microarray by the order of interaction cascades. A systems approach was developed in this study by linking differentially expressed genes with publicly available PPI data, Gene Ontology and subcellular localizaton for the integrated analyses. These shortest paths from NGAL to DEG transcription factors or other transcription factors in the PPI network provide important clues for future experimental identification of new pathways.
Related JoVE Video
Genetic variants in the KDR gene is associated with the prognosis of transarterial chemoembolization treated hepatocellular carcinoma.
Tumour Biol.
PUBLISHED: 08-16-2014
Show Abstract
Hide Abstract
Kinase insert domain receptor (KDR) is the principal receptor that promotes the proangiogenic action of vascular endothelial growth factor and is involved in the tumorigenesis and progression of many malignancies, including hepatocellular carcinoma (HCC). Single-nucleotide polymorphisms (SNPs) of KDR have been reported to be with the risk and prognosis of several malignancies. Our aim was to determine whether SNPs in KDR gene are associated with clinical outcomes in HCC patients treated with transcatheter arterial chemoembolization. A total of 192 HCC patients were tested for KDR SNPs, and the SNP results were correlated with progression-free survival (PFS) and overall survival (OS). The association of the SNPs with the overall survival (OS) of patients was assessed by Kaplan-Meier method, and then Cox proportional hazards model was used to assess the variables resulted significant at univariate analysis. No significant differences were found in correlation between KDR SNPs and patients' PFS. Our data showed that genotype AA?+?TA of rs1870377 and genotype CC?+?TC of rs2071559 were significantly associated with overall survival of HCC patients (P?400 ?g/L), existence of portal vein tumor thrombus, and high BCLC stage (HR?=?0.61; 95 % CI, 0.36-0.88; P?=?0.003 and HR?=?0.54; 95 % CI, 0.40-0.94; P?=?0.002, respectively). Our results suggest that SNPs rs1870377 and rs2071559 in the KDR gene may serve as independent prognosis biomarkers for unresectable HCC patient, which warranted further validating investigation.
Related JoVE Video
A new, quick, highly sensitive ultramicro-analysis method for the identification of fructose removed from fructofuranosyl-containing gluco-oligosaccharides by ESI-CID-MS/MS.
Carbohydr. Res.
PUBLISHED: 08-04-2014
Show Abstract
Hide Abstract
An efficient, highly sensitive, and ultramicroscale analytical method for the identification of fructose removed from fructofuranosyl-containing gluco-oligosaccharides, including malto-oligosyl fructofuranosides and oligomeric (1?2)-?-d-glucopyranosyl-(1?2)-?-d-fructofuranosides by ESI-CID-MS/MS has been developed with proven applications far superior to the existing method using NMR. With the established principle of diagnostic fragmentation by ESI-CID-MS/MS, the terminal saccharide (either glucose or fructose) can be readily and unambiguously determined at high sensitivity without a tedious derivatization process. Detection of the A-type fragmentation (0,4)A-h type ion, and (0,2)A type ion are useful as a diagnostic fragmentation tool to identify whether fructose terminal is removed from oligosaccharides. It will facilitate the efficient production of suitable oligosaccharide microarrays crucial for studies on carbohydrate-protein interaction in seeking functional carbohydrates.
Related JoVE Video
Sorafenib continuation after first disease progression could reduce disease flares and provide survival benefits in patients with hepatocellular carcinoma: a pilot retrospective study.
Asian Pac. J. Cancer Prev.
PUBLISHED: 05-13-2014
Show Abstract
Hide Abstract
Sorafenib is a promising drug for advanced hepatocellular carcinoma (HCC); however, treatment may be discontinued for multiple reasons, such as progressive disease, adverse events, or the cost of treatment. The consequences of sorafenib discontinuation and continuation are uncertain.
Related JoVE Video
[Zoonotic trematode species identified in domestic animals from Huainan area].
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi
PUBLISHED: 05-08-2014
Show Abstract
Hide Abstract
To understand the zoonotic trematode species identified in domestic animals from Huainan area, Anhui Province, so as to supply the evidence for the prevention of zoonoses.
Related JoVE Video
Worsening epidemic of HIV and syphilis among men who have sex with men in Jiangsu Province, China.
Clin. Infect. Dis.
PUBLISHED: 03-18-2014
Show Abstract
Hide Abstract
Epidemics of human immunodeficiency virus (HIV) and syphilis among men who have sex with men (MSM) are major public health concerns in most parts of China. A dearth of information regarding the current trend of HIV in this hard-to-reach population in several regions including Jiangsu, coupled with the reemergence of syphilis, calls for an effort to understand the dynamics of the dual epidemic in this province.
Related JoVE Video
Identification and validation co-differentially expressed genes with NAFLD and insulin resistance.
Endocrine
PUBLISHED: 03-14-2014
Show Abstract
Hide Abstract
Insulin resistance is a major feature and pathogenic factor of nonalcoholic fatty liver disease (NAFLD). Theoretically, genetic variation in candidate genes related to insulin resistance may contribute to the pathogenesis of NAFLD. The purpose of this study was to identify potentially pathogenic genes involved in NAFLD and insulin resistance that have not yet been discovered. This study yielded five important discoveries. 1. A total of 21 co-differentially expressed genes in both the NAFLD and insulin-resistance groups were identified from the pool containing thousands of genes via the significance analysis of microarrays method. 2. MAP kinase-interacting serine/threonine kinase 2 (Mknk2) was the unique gene to be identified that is involved in the insulin signaling pathway and Mitogen Activated Protein Kinase signaling pathway according to the Kyoto Encyclopedia of Genes and Genomes database. 3. Mknk2 mRNA and protein expression were dose-dependently up-regulated by palmitic acid (PA) in mouse primary hepatocytes. 4. Western blotting analysis and quantitative real-time PCR confirmed that Mknk2 affected the expression of acetyl-CoA carboxylases-1 and fatty acid synthase. 5. The inhibition of Mknk2 alleviated PA-induced insulin resistance, whereas the overexpression of Mknk2 resulted in the aggravation of insulin resistance in PA-treated hepatocytes. Therefore, we predict that MKNK2 may be a key protein related to NAFLD and insulin resistance.
Related JoVE Video
Rapid screening for sickle cell disease by polymerase chain reaction-high resolution melting analysis.
Mol Med Rep
PUBLISHED: 03-05-2014
Show Abstract
Hide Abstract
Each year, ~300,000 individuals with sickle cell disease (SCD), a hemoglobinopathy caused by ?-globin gene mutation, are born, and >75% of those are in Africa. The present study examined 511 individuals on the island of Bioko (Equatorial Guinea) and attempted to establish a method for rapid sickle cell disease screening. Following DNA extraction and polymerase chain reaction (PCR) amplification, high resolution melting (HRM) analysis was used to assess the specificity of fluorescence signals of the PCR products and to differentiate various genotypes of these products. The analytical results of HRM were validated using DNA sequencing. By HRM analysis, 80 out of 511 samples were classified as hemoglobin S (Hb S) heterozygotes, while 431 out of 511 samples were classified as wild-type. No mutant homozygote was identified. DNA sequencing indicated that within the 431 wild-type samples as indicated by HRM analysis, one case was actually a Hb S heterozygote and another case was a rare hemoglobin S-C genotype (sickle-hemoglobin C disease). One out of 80 suspected Hb S heterozygotes as indicated by HRM was confirmed as wild-type by DNA sequencing and the results of residual 508 cases were consistent for HRM analysis and sequencing. In conclusion, HRM analysis is a simple, high-efficiency approach for Hb S screening and is useful for early diagnosis of SCD and particularly suitable for application in the African area.
Related JoVE Video
Retracted:EEG Synchronization Evaluation: A New Diagnostic Tool for Predicting the Progression of Alzheimer's disease.
Clin EEG Neurosci
PUBLISHED: 03-03-2014
Show Abstract
Hide Abstract
"EEG Synchronization Evaluation: A New Diagnostic Tool for Predicting the Progression of Alzheimer's Disease" by Ran Yi, Xiao-rong Zhan, Jing Tang, Li-ming Zhang, Xiao-min Liu and Qi Dong, Clin EEG Neurosci published online 3 March 2014 (DOI: 10.1177/1550059413502779) This article has been retracted due to unattributed inclusion of significant excerpts from the following article: Knyazeva, Maria G.; Carmeli, Cristian; Khadivi, Alireza; Ghika, Joseph; Meuli, Reto; Frackowiak, Richard S. (2013). "Evolution of source EEG synchronization in early Alzheimer's disease". Neurobiology of Aging, 34(3), 694-705. Alzheimer's disease (AD) is known as a leading cause of dementia in elderly persons. It is a chronic neurodegenerative disorder characterized by progressive cognitive dysfunction. AD can disrupt functional connectivity in distributed cortical networks. The S-estimator, which is a measure of multivariate intraregional synchronization, was analyzed in this study. Twenty patients with AD and 20 age-matched controls were tested at baseline and after 1 year to evaluate the potential of synchronization to be a possible marker of AD progression. All the subjects had clinical evaluations and electroencephalography (EEG) at baseline and post 1 year. Hyposynchronization had an important effect in the medial temporal and frontal regions, while there were no significant effects for hypersynchronization. Hypersynchronized clusters changed more slowly with time (P = .067), whereas hyposynchronized clusters changed more quickly (P = .032). Hyposynchronized cluster-averaged S-estimator correlated negatively with progression of AD (r = -0.98769, P = .0103). In conclusion, the present study provides a whole-brain, AD-specific phenotype of temporal coordination in distributed cortical networks, which is an early diagnostic tool for progression of AD.
Related JoVE Video
Novel integration strategy for enhancing chalcopyrite bioleaching by Acidithiobacillus sp. in a 7-L fermenter.
Bioresour. Technol.
PUBLISHED: 01-23-2014
Show Abstract
Hide Abstract
An integrated strategy (additional energy substrate-three stage pH control-fed batch) was firstly proposed for efficiently improving chalcopyrite bioleaching by Acidithiobacillus sp. in a 7-L fermenter. The strain adaptive-growing phase was greatly shortened from 8days into 4days with the supplement of additional 2g/L Fe(2+)+2g/L S(0). Jarosite passivation was effectively weakened basing on higher biomass via the three-stage pH-stat control (pH 1.3-1.0-0.7). The mineral substrate inhibition was attenuated by fed-batch fermentation. With the integrated strategy, the biochemical reaction was promoted and achieved a better balance. Meanwhile, the domination course of A. thiooxidans in the microbial community was shortened from 14days to 8days. As the results of integrated strategy, the final copper ion and productivity reached 89.1mg/L and 2.23mg/(Ld), respectively, which was improved by 52.8% compared to the uncontrolled batch bioleaching. The integrated strategy could be further exploited for industrial chalcopyrite bioleaching.
Related JoVE Video
Laparoscopic splenectomy for hypersplenism secondary to liver cirrhosis and portal hypertension.
World J. Gastroenterol.
PUBLISHED: 01-13-2014
Show Abstract
Hide Abstract
Since the first laparoscopic splenectomy (LS) was reported in 1991, LS has become the gold standard for the removal of normal to moderately enlarged spleens in benign conditions. Compared with open splenectomy, fewer postsurgical complications and better postoperative recovery have been observed, but LS is contraindicated for hypersplenism secondary to liver cirrhosis in many institutions owing to technical difficulties associated with splenomegaly, well-developed collateral circulation, and increased risk of bleeding. With the improvements of laparoscopic technique, the concept is changing. This article aims to give an overview of the latest development in laparoscopic splenectomy for hypersplenism secondary to liver cirrhosis and portal hypertension. Despite a lack of randomized controlled trial, the publications obtained have shown that with meticulous surgical techniques and advanced instruments, LS is a technically feasible, safe, and effective procedure for hypersplenism secondary to cirrhosis and portal hypertension and contributes to decreased blood loss, shorter hospital stay, and less impairment of liver function. It is recommended that the dilated short gastric vessels and other enlarged collateral circulation surrounding the spleen be divided with the LigaSure vessel sealing equipment, and the splenic artery and vein be transected en bloc with the application of the endovascular stapler. To support the clinical evidence, further randomized controlled trials about this topic are necessary.
Related JoVE Video
High resolution melting analysis: a rapid screening and typing tool for common ?-thalassemia mutation in Chinese population.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
?-thalassemia is a common inherited disorder worldwide including southern China, and at least 45 distinct ?-thalassemia mutations have been identified in China. High-resolution melting (HRM) assay was recently introduced as a rapid, inexpensive and effective method for genotyping. However, there was no systemic study on the diagnostic capability of HRM to identify ?-thalassemia. Here, we used an improved HRM method to screen and type 12 common ?-thalassemia mutations in Chinese, and the rapidity and reliability of this method was investigated. The whole PCR and HRM procedure could be completed in 40 min. The heterozygous mutations and 4 kinds of homozygous mutations could be readily differentiated from the melting curve except c.-78A>G heterozygote and c.-79A>G heterozygote. The diagnostic reliability of this HRM assay was evaluated on 756 pre-typed genomic DNA samples and 50 cases of blood spots on filter paper, which were collected from seven high prevalent provinces in southern China. If c.-78A>G heterozygote and c.-79A>G heterozygote were classified into the same group (c.-78&79 A>G heterozygote), the HRM method was in complete concordance with the reference method (reverse dot blot/DNA-sequencing). In a conclusion, the HRM method appears to be an accurate and sensitive method for the rapid screening and identification of ?-thalassemia mutations. In the future, we suggest this technology to be used in neonatal blood spot screening program. It could enlarge the coverage of ?-thalassemia screening program in China. At the same time, its value should be confirmed in prospectively clinical and epidemiological studies.
Related JoVE Video
Acaroid mite allergens from the filters of air-conditioning system in China.
Int J Clin Exp Med
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Accumulation of acaroid mites in the filters of air-conditioners is harmful to human health. It is important to clarify the allergen components of mites from the filters of local air-conditioning system. The present study was to detect the allergen types in the filters of air-conditioners and assesse their allergenicity by asthmatic models. Sixty aliquots of dust samples were collected from air conditioning filters in civil houses in Wuhu area. Total protein was extracted from the dust samples using PBS and quantified by Bradford method. Allergens I and II were also detected by Western blot using primary antibody (anti-Der f1/2, Der p1/Der f2/Der p2, respectively). Ten aliquots of the positive samples were randomly selected for homogenization and sensitized the mice for developing asthmatic animal models. Total serum IgE level and IFN-?, IL-4 and IL-5 in the bronchoalveolar lavage fluid (BALF). The allergenicity of the extraction was assessed using pathological sections developed from the mouse pulmonary tissues. The concentration of extract from the 60 samples was ranged from 4.37 ?g/ml to 30.76 ?g/ml. After analyzing with Western blot, 31 of 60 samples were positive for 4 allergens of acaroid mites, and yet 16 were negative. The levels of total IgE from serum IL-4 and IL-5 from the BALF in the experimental group were apparently higher than that of negative control and PBS group (P < 0.01), but there were no statistical difference compared to OVA group (P > 0.05). However,the IFN-? level in BALF was lower compared with the negative control and PBS group (P < 0.05) but with the OVA group (P > 0.05). The pathological changes were evidently emerged in pulmonary tissues, which were similar to those of OVA group, compared with the PBS ground and negative controls. The air-conditioner filters in human dwellings of Wuhu area potentially contain the major group allergen 1 and 2 from D. farinae and D. pteronyssinus, which may be associated with seasonal prevalence of allergic disorders in this area.
Related JoVE Video
Molecular epidemiological characterization and health burden of thalassemia in Jiangxi Province, P. R. China.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Thalassemia is the most common inherited disease in southern China. However, this disorder is usually ignored by Jiangxi provincial health system and government due to lack of epidemiological data.
Related JoVE Video
Curdlan ?-1,3-glucooligosaccharides induce the defense responses against phytophthora infestans infection of potato (Solanum tuberosum L. cv. McCain G1) leaf cells.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Activation of the innate immune system before the invasion of pathogens is a promising way to improve the resistance of plant against infection while reducing the use of agricultural chemicals. Although several elicitors were used to induce the resistance of potato plant to microbial pathogen infection, the role of curdlan oligosaccharide (CurdO) has not been established. In the current study, the defense responses were investigated at biochemical and proteomic levels to elucidate the elicitation effect of CurdOs in foliar tissues of potato (Solanum tuberosum L. cv. McCain G1). The results indicate that the CurdOs exhibit activation effect on the early- and late-defense responses in potato leaves. In addition, glucopentaose was proved to be the shortest active curdlan molecule based on the accumulation of H?O? and salicylic acid and the activities of phenylalanine amino-lyase, ?-1,3-glucanase and chitinase. The 2D-PAGE analysis reveals that CurdOs activate the integrated response reactions in potato cells, as a number of proteins with various functions are up-regulated including disease/defense, metabolism, transcription, and cell structure. The pathogenesis assay shows that the ratio of lesion area of potato leaf decreased from 15.82%±5.44% to 7.79%±3.03% when the plants were treated with CurdOs 1 day before the infection of Phytophthora infestans. Furthermore, the results on potato yield and induction reactions indicate that the defense responses induced by CurdOs lasted for short period of time but disappeared gradually.
Related JoVE Video
Transcriptome sequencing and gene expression analysis of Trichoderma brevicompactum under different culture conditions.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Trichoderma brevicompactum is the Trichoderma species producing simple trichothecenes-trichodermin, a potential antifungal antibiotic and a protein synthesis inhibitor. However, the biosynthetic pathway of trichodermin in Trichoderma is not completely clarified. Therefore, transcriptome and gene expression profiling data for this species are needed as an important resource to better understand the mechanism of the trichodermin biosynthesis and provide a blueprint for further study of T. brevicompactum.
Related JoVE Video
[Preliminary studies on protective immunity induced by recombinant protein of Schistosoma japonicum P14 gene and its mechanism against schistosome infection in mice].
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi
PUBLISHED: 12-24-2013
Show Abstract
Hide Abstract
To explore the immune protective effect of the basic calponin-like protein (SjP14) in mice response to the infection of Schistosoma japonicum.
Related JoVE Video
[Expression of serum microRNAs (miR-222, miR-181, miR-216) in human hepatocellular carcinoma and its clinical significance].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 10-16-2013
Show Abstract
Hide Abstract
To investigate the abnormal expression of microRNAs (miR-216, miR-222, miR-181) in the serum of patients with hepatocellular carcinoma (HCC) and its clinical significance.
Related JoVE Video
A novel osmotic pressure control fed-batch fermentation strategy for improvement of erythritol production by Yarrowia lipolytica from glycerol.
Bioresour. Technol.
PUBLISHED: 08-26-2013
Show Abstract
Hide Abstract
The effect of osmotic pressure on erythritol and mannitol production by an osmophilic yeast strain of Yarrowia lipolytica CICC 1675 using glycerol as the sole carbon source was investigated. Appropriately high osmotic pressure was found to enhance erythritol production and inhibit mannitol formation. A novel two-stage osmotic pressure control fed-batch strategy based on the kinetic analysis was developed for higher erythritol yield and productivity. During the first 96h, the osmotic pressure was maintained at 4.25osmol/kg by feeding glycerol to reduce the inhibition of cell growth. After 132h, the osmotic pressure was controlled at 4.94osmol/kg to maintain a high dpery/dt. Maximum erythritol yield of 194.3g/L was obtained with 0.95g/L/h productivity, which were 25.7% and 2.2%, respectively, improvement over the best results in one-stage fed-batch fermentation. This is the first report that a novel osmotic pressure control fed-batch strategy significantly enhanced erythritol production.
Related JoVE Video
Effect of prolonged weekly hemodialysis on survival of maintenance hemodialysis patients: a meta-analysis of studies.
Nephron Clin Pract
PUBLISHED: 07-24-2013
Show Abstract
Hide Abstract
Objective: Use of prolonged nocturnal or daytime hemodialysis (PHD, more than 12 h per week) is associated with improvement of some clinical parameters relative to conventional hemodialysis (CHD, 4 h sessions, thrice weekly), but the effect on survival is unclear. The purpose of this meta-analysis is to determine whether PHD improves survival of patients undergoing maintenance HD. Design: Systematic review of observational studies by meta-analysis. Data Sources: Electronic searches in MEDLINE (PubMed, 1966-2012), EMBASE (1974-2012), www.clinicaltrials.gov, and the Cochrane Controlled Clinical Trials Register Database. Eligibility Criteria for Selecting Studies: All prospective or retrospective studies were considered eligible if they were cohort studies or observational studies that compared CHD with PHD (more than 12 h of HD per week due to more HD sessions or increased duration of HD sessions) and the final outcome was all-cause death or mortality. Results: Thirteen studies with a total of 85,722 participants (10,285 PHD patients, 75,437 CHD patients) met the inclusion criteria. Summary estimates indicated that PHD was associated with decreased risk of mortality (OR = 0.72, 95% CI 0.64-0.81, p < 0.00001). Analysis of residual confounders of pooled results from six retrospective studies indicated that PHD patients were less likely to have low hemoglobin (11.7 vs. 11.2 g/dl, p < 0.01), younger (51.2 vs. 58.8 years, p < 0.01), less likely to have diabetes (27.1 vs. 40.8%, p < 0.01), and less likely to use a catheter (18.4 vs. 27.1%, p < 0.01), so these may have affected the outcome measure in these studies. Conclusions: PHD is associated with improved survival relative to CHD, although residual confounders have affected this relationship in observational studies. Large, multicenter randomized, controlled trials are needed to confirm our results. © 2013 S. Karger AG, Basel.
Related JoVE Video
Molecular epidemiological survey of hemoglobinopathies in the Wuxi region of Jiangsu Province, eastern China.
Hemoglobin
PUBLISHED: 06-27-2013
Show Abstract
Hide Abstract
In order to determine the prevalence and molecular characterization of hemoglobinopathies in the Wuxi region of Jiangsu Province in the Peoples Republic of China (PRC), a total of 10,297 healthy people selected from a regional hospital were screened. Hemoglobin (Hb) electrophoresis, complete blood cell (CBC) count, polymerase chain reaction (PCR), DNA sequencing, reverse dot-blot and multiplex ligation-dependent probe amplification (MLPA) were used to detect Hb variants, thalassemias and hereditary persistence of fetal Hb (HPFH). Two thousand and twenty-one adult subjects were screened for thalassemia, five cases were identified as ?-thalassemia (?-thal) carriers including three cases of the -?(3.7) (rightward) deletion, one case of the - -(SEA) deletion and one case of ?-thal [IVS-II-654 (C>T), (HBB: c.316-197C>T)]. The incidence of Hb variants, thalassemia and HPFH/??-thal were 0.136% (14/10,297), 0.25% (5/2021) and 0.0001% (1/10,297), respectively. Eight genotypes of Hb variants were found, including Hb E [?26(B8)Glu?Lys, GAG>AAG; HBB: c.79G>A], Hb J-Bangkok [?56(D7)Gly?Asp (GGC>GAC); HBB; c.170G>A], Hb G-Coushatta [?22(4)Glu?Ala (GAA>GCA); HBB: c.68A>C], Hb Queens [?34(B15)Leu?Arg (CTG>CGG) (?2 or ?1); HBA2: c.104T>G (or HBA1)], Hb I [?16(A14)Lys?Glu, AAG>GAG (?1); HBA1: c.49A>G], Hb Beijing [?16(A14)Lys?Asn (AAG>AAC or AAT) (?2 or ?1); HBA2: c.51G>C (or HBA1) or 51G>T (or HBA1)], Hb Ube-2 [?68(E17)Asn?Asp (AAC>GAC) (?2 or ?1); HBA2: c.205A>G (or HBA1)] and Hb G-Taipei [?22(B4)Glu?Gly (GAA>GGA); HBB: c.68A>G]. A Sicilian ??(0)-thal, identified for the first time in Asia, was also found in this survey.
Related JoVE Video
A new effective process for production of curdlan oligosaccharides based on alkali-neutralization treatment and acid hydrolysis of curdlan particles in water suspension.
Appl. Microbiol. Biotechnol.
PUBLISHED: 05-20-2013
Show Abstract
Hide Abstract
Biologically active ?-1,3-oligosaccharides with rapidly growing biomedical applications are produced from hydrolysis of curdlan polysaccharide. The water-insoluble curdlan impedes its hydrolysis efficiency which is enhanced by our newly developed alkali-neutralization treatment process to increase the stability of curdlan suspension to more than 20 days, while the untreated control settled within 5 min. A putative double-layer structure model comprising of a compact core and a hydrated outer layer was proposed to describe the treated curdlan particles based on sedimentation and scanning electron microscopy observation. This model was verified by single- and two-step acid hydrolysis, indicative of the reduced susceptibility to hydrolysis when close to the compact core. Electrospray ionization-mass spectrometry, thin-layer chromatography analyses, and effective HPLC procedure led to the development of improved process to produce purified individual ?-1,3-oligosaccharides with degrees of polymerization from 2 to 10 and potential for biomedical applications from curdlan hydrolyzate. Our new curdlan oligosaccharide production process offers an even better alternative to the previously published processes.
Related JoVE Video
Low prevalence of HPV in male sexual partners of HR-HPV infected females and low concordance of viral types in couples in Eastern Guangdong.
Asian Pac. J. Cancer Prev.
PUBLISHED: 05-18-2013
Show Abstract
Hide Abstract
To investigate the prevalence of genital high-risk human papillomavirus (HR-HPV) in male sexual partners of HR-HPV infected women and the concordance of viral types in couples in China, and comprehend the role of men play in HPV transmission to women.
Related JoVE Video
[Prognostic significance of serum level of vascular endothelial growth factor receptor-2 in hepatocellular carcinoma patients after transcatheter arterial chemoembolization].
Zhonghua Yi Xue Za Zhi
PUBLISHED: 05-11-2013
Show Abstract
Hide Abstract
To explore the prognostic significance of serum vascular endothelial growth factor receptor-2 (VEGFR-2) in patients with hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE).
Related JoVE Video
An open-labeled, randomized, multicenter phase IIa study of gambogic acid injection for advanced malignant tumors.
Chin. Med. J.
PUBLISHED: 05-09-2013
Show Abstract
Hide Abstract
Gambogic acid is a pure active compound isolated from the traditional Chinese medicinal plant gamboge (Garcinia morella Desv.). Based on the preliminary results of a phase I study, this phase IIa study compared the efficacy and safety of different dosage schedules of gambogic acid in patients with advanced malignant tumors.
Related JoVE Video
Multivalent scaffold proteins as superagonists of TRAIL receptor 2-induced apoptosis.
Mol. Cancer Ther.
PUBLISHED: 05-03-2013
Show Abstract
Hide Abstract
Activation of TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2) can induce apoptosis in a variety of human cancer cell lines and xenografts, while lacking toxicity in normal cells. The natural ligand and agonistic antibodies show antitumor activity in preclinical models of cancer, and this had led to significant excitement in the clinical potential of these agents. Unfortunately, this optimism has been tempered by trial data that, thus far, are not showing clear signs of efficacy in cancer patients. The reasons for discrepant preclinical and clinical observations are not understood, but one possibility is that the current TRAILR2 agonists lack sufficient potency to achieve a meaningful response in patients. Toward addressing that possibility, we have developed multivalent forms of a new binding scaffold (Tn3) that are superagonists of TRAILR2 and can induce apoptosis in tumor cell lines at subpicomolar concentrations. The monomer Tn3 unit was a fibronectin type III domain engineered for high-affinity TRAILR2 binding. Multivalent presentation of this basic unit induced cell death in TRAILR2-expressing cell lines. Optimization of binding affinity, molecular format, and valency contributed to cumulative enhancements of agonistic activity. An optimized multivalent agonist consisting of 8 tandem Tn3 repeats was highly potent in triggering cell death in TRAIL-sensitive cell lines and was 1 to 2 orders of magnitude more potent than TRAIL. Enhanced potency was also observed in vivo in a tumor xenograft setting. The TRAILR2 superagonists described here have the potential for superior clinical activity in settings insensitive to the current therapeutic agonists that target this pathway.
Related JoVE Video
All-trans-retinoic acid ameliorates experimental allergic encephalomyelitis by affecting dendritic cell and monocyte development.
Immunology
PUBLISHED: 04-27-2013
Show Abstract
Hide Abstract
Experimental allergic encephalomyelitis (EAE) can be induced in animal models by injecting the MOG35-55 peptide subcutaneously. Dendritic cells (DCs) that are located at the immunization site phagocytose the MOG35-55 peptide. These DCs mature and migrate into the nearest draining lymph nodes (dLNs), then present antigen, resulting in the activation of naive T cells. T helper type 1 (Th1) and Th17 cells are the primary cells involved in EAE progression. All-trans-retinoic acid (AT-RA) has been shown to have beneficial effects on EAE progression; however, whether AT-RA influences DC maturation or mediates other functions is unclear. In the present study, we showed that AT-RA led to the down-regulation of MHC class II, CD80 (B7-1) and CD86 (B7-2) expressed on the surface of DCs that were isolated from dLNs or spleen 3 days post-immunization in an EAE model. Changes to DC function influenced Th1/Th17 subset polarization. Furthermore, the number of CD44(+) monocytes (which might trigger EAE progression) was also significantly decreased in dLNs, spleen, subarachnoid space and the spinal cord parenchyma after AT-RA treatment. These findings are the first to demonstrate that AT-RA impairs the antigen-presenting capacity of DCs, leading to down-regulation of pathogenic Th1 and Th17 inflammatory cell responses and reducing EAE severity.
Related JoVE Video
Locating QTLs controlling several adult root traits in an elite Chinese hybrid rice.
Gene
PUBLISHED: 04-11-2013
Show Abstract
Hide Abstract
This study aimed to elucidate the genetics of the adult root system in elite Chinese hybrid rice. Several adult root traits in a recombinant inbred line (RIL) population of Xieyou 9308 and two backcross F1 (BCF1) populations derived from the RILs were phenotyped under hydroponic culture at heading stage for quantitative trait locus (QTL) mapping and other statistical analysis. There a total of eight QTLs detected for the root traits. Among of them, a pleiotropic QTL was repeatedly flanked by RM180 and RM5436 on the short arm of chromosome 7 for multiple traits across RILs and its BCF1 populations, accounting for 6.88% to 25.26% of the phenotypic variances. Only additive/dominant QTLs were detected for the root traits. These results can serve as a foundation for facilitating future cloning and molecular breeding.
Related JoVE Video
Enhancing pIFN-? production and process stability in fed-batch culture of pichia pastoris by controlling the methanol concentration and monitoring the responses of OUR/DO levels.
Appl. Biochem. Biotechnol.
PUBLISHED: 04-03-2013
Show Abstract
Hide Abstract
Effective expression of porcine interferon-? (pIFN-?) with recombinant Pichia pastoris was conducted in a bench-scale fermentor using an in situ methanol electrode-based feeding process with the control level of methanol concentration linearly increased to 10 g l?¹ for the first 20 h and maintained at 10 g l?¹ for the rest of expression phase. With this two-stage control process, the highest pIFN-? concentration reached a level of 1.81 g l?¹, which was 1.5-fold of that in the previous constant 10 g l?¹ induction experiments. There is an improvement of the pIFN-? productivity from more distribution of carbon flux to protein expression. The pIFN-? expression stability could be further enhanced by a simple on-line fault diagnosis method for methanol overfeeding based on oxygen uptake rate changing patterns. By implementing corrective action of feeding glycerol after fault detection, the production yield increased to twice the amount it would have been without the diagnosis.
Related JoVE Video
A?40 modulates GABAA receptor ?6 subunit expression and rat cerebellar granule neuron maturation through the ERK/mTOR pathway.
J. Neurochem.
PUBLISHED: 03-28-2013
Show Abstract
Hide Abstract
In addition to their neurotoxic role in Alzheimers disease (AD), ?-amyloid peptides (A?s) are also known to play physiological roles. Here, we show that recombinantA?40 significantly increased the outward current of the GABAA receptor containing(GABAA ?6) in rat cerebellar granule neurons (CGNs). The A?40-mediated increase in GABAA ?6 current was mediated by an increase in GABAA ?6 protein expression at the translational rather than the transcriptional level. The exposure of CGNs to A?40 markedly induced the phosphorylation of ERK (pERK) and mammalian target of rapamycin (pmTOR). The increase in GABAA ?6 current and expression was attenuated by specific inhibitors of ERK or mTOR, suggesting that the ERK and mTOR signaling pathways are required for the effect of A?40 on GABAA ?6 current and expression in CGNs. A pharmacological blockade of the p75 neurotrophin receptor (p75(NTR) ), but not the insulin or ?7-nAChR receptors, abrogated the effect of A?40 on GABAA ?6 protein expression and current. Furthermore, the expression of GABAA ?6 was lower in CGNs from APP(-/-) mice than in CGNs from wild-type mice. Moreover, the internal granule layer (IGL) in APP(-/-) mice was thinner than the IGL in wild-type mice. The injection of A?40 into the cerebellum reversed this effect, and the application of p75(NTR) blocking antibody abolished the effects of A?40 on cerebellum morphology in APP(-/-) mice. Our results suggest that low concentrations of A?40 play a role in regulating CGN maturation through p75(NTR) . This article is protected by copyright. All rights reserved.
Related JoVE Video
Hemoglobinopathy: molecular epidemiological characteristics and health effects on Hakka people in the Meizhou region, southern China.
PLoS ONE
PUBLISHED: 02-01-2013
Show Abstract
Hide Abstract
Hemoglobinopathies are the most common inherited diseases in southern China. However, there have been only a few epidemiological studies of hemoglobinopathies in Guangdong province.
Related JoVE Video
Progesterone alleviates neural behavioral deficits and demyelination with reduced degeneration of oligodendroglial cells in cuprizone-induced mice.
PLoS ONE
PUBLISHED: 01-24-2013
Show Abstract
Hide Abstract
Demyelination occurs widely in neurodegenerative diseases. Progesterone has neuroprotective effects, is known to reduce the clinical scores and the inflammatory response. Progesterone also promotes remyelination in experimental autoimmune encephalomyelitis and cuprizone-induced demyelinating brain. However, it still remains unclear whether progesterone can alleviate neural behavioral deficits and demyelination with degeneration of oligodendroglial cells in cuprizone-induced mice. In this study, mice were fed with 0.2% cuprizone to induce demyelination, and treated with progesterone to test its potential protective effect on neural behavioral deficits, demyelination and degeneration of oligodendroglial cells. Our results showed noticeable alleviation of neural behavioral deficits following progesterone treatment as assessed by changes in average body weight, and activity during the open field and Rota-rod tests when compared with the vehicle treated cuprizone group. Progesterone treatment alleviated demyelination as shown by Luxol fast blue staining, MBP immunohistochemical staining, and electron microscopy. There was an obvious decrease in TUNEL and Caspase-3-positive apoptotic cells, and an increase in the number of oligodendroglial cells staining positive for PDGFR?, Olig2, Sox10 and CC-1 antibody in the brains of cuprizone-induced mice after progesterone administration. These results indicate that progesterone can alleviate neural behavioral deficits and demyelination against oligodendroglial cell degeneration in cuprizone-induced mice.
Related JoVE Video
Exposure to extremely low-frequency electromagnetic fields modulates Na+ currents in rat cerebellar granule cells through increase of AA/PGE2 and EP receptor-mediated cAMP/PKA pathway.
PLoS ONE
PUBLISHED: 01-22-2013
Show Abstract
Hide Abstract
Although the modulation of Ca(2+) channel activity by extremely low-frequency electromagnetic fields (ELF-EMF) has been studied previously, few reports have addressed the effects of such fields on the activity of voltage-activated Na(+) channels (Na(v)). Here, we investigated the effects of ELF-EMF on Na(v) activity in rat cerebellar granule cells (GCs). Our results reveal that exposing cerebellar GCs to ELF-EMF for 10-60 min significantly increased Na(v) currents (I(Na)) by 30-125% in a time- and intensity-dependent manner. The Na(v) channel steady-state activation curve, but not the steady-state inactivation curve, was significantly shifted (by 5.2 mV) towards hyperpolarization by ELF-EMF stimulation. This phenomenon is similar to the effect of intracellular application of arachidonic acid (AA) and prostaglandin E(2) (PGE(2)) on I(Na) in cerebellar GCs. Increases in intracellular AA, PGE(2) and phosphorylated PKA levels in cerebellar GCs were observed following ELF-EMF exposure. Western blottings indicated that the Na(V) 1.2 protein on the cerebellar GCs membrane was increased, the total expression levels of Na(V) 1.2 protein were not affected after exposure to ELF-EMF. Cyclooxygenase inhibitors and PGE(2) receptor (EP) antagonists were able to eliminate this ELF-EMF-induced increase in phosphorylated PKA and I(Na). In addition, ELF-EMF exposure significantly enhanced the activity of PLA(2) in cerebellar GCs but did not affect COX-1 or COX-2 activity. Together, these data demonstrate for the first time that neuronal I(Na) is significantly increased by ELF-EMF exposure via a cPLA2 AA PGE(2) EP receptors PKA signaling pathway.
Related JoVE Video
The Elicitation Effect of Pathogenic Fungi on Trichodermin Production by Trichoderma brevicompactum.
ScientificWorldJournal
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
The effects of six species of phytopathogenic fungi mycelia as elicitors on trichodermin yield by Trichoderma brevicompactum were investigated. Neither nonviable nor viable mycelia of Botrytis cinerea, Alternaria solani, Colletotrichum lindemuthianum, and Thanatephorus cucumeris demonstrated any elicitation on the accumulation of trichodermin. However, the production of trichodermin was increased by the presence of viable/nonviable Rhizoctonia solani and Fusarium oxysporum mycelia. The strongest elicitation effect was found at the presence of nonviable R. solani. At the presence of nonviable R. solani, the maximum yield of trichodermin (144.55?mg/L) was significantly higher than the Control (67.8?mg/L), and the cultivation time to obtain the maximum yield of trichodermin decreased from 72?h to 60?h. No difference of trichodermin accumulation was observed by changing the concentration of nonviable R. solani from 0.1 to 1.6?g/L. It was observed that the optimum time for adding nonviable R. solani is immediately after inoculation. The diameter of T. brevicompactum mycelial globule after 72?h cultivation with nonviable R. solani elicitor was smaller than that of the Control.
Related JoVE Video
[Pathological changes in rat model of urinary calculus induced by melamine].
Zhonghua Er Ke Za Zhi
PUBLISHED: 12-20-2011
Show Abstract
Hide Abstract
To investigate melamine-induced pathological changes in the kidney.
Related JoVE Video
Development and evaluation of a reverse dot blot assay for the simultaneous detection of common alpha and beta thalassemia in Chinese.
Blood Cells Mol. Dis.
PUBLISHED: 11-04-2011
Show Abstract
Hide Abstract
Thalassemia is the commonest inherited autosomal recessive disorders of hemoglobin in southern China. We developed and evaluated a reverse dot blot (RDB) assay combined with flow-through hybridization technology platform for the rapid and simultaneous identification of 5 types of ?-thalassemia and 16 types of ?-thalassemia common in Chinese. Reliable genotyping of wild-type and thalassemic genomic DNA samples was achieved by means of a gene chip on which allele-specific oligonucleotide probes were immobilized on a nylon membrane. This method involved two multiplex PCR amplification systems of ?-thalassemia and ?-thalassemia and one time of hybridization. The whole procedure starting from blood sampling to the identification of thalassemia genotype required less than 4h. The diagnostic reliability of this reverse dot blot assay was evaluated on 427 samples (387 cases of thalassemia and 40 healthy persons) by using direct DNA sequence analysis and gap-PCR in a blind study. These samples included 377 cases of blood, 7 cases of amniotic fluid, 18 cases of chorionic villus, and 25 cases of cord blood. The RDB gene chip was in complete concordance with the reference method. The reverse dot blot assay was a simple, rapid, accurate, and cost-effective method to identify common thalassemia genotypes in the Chinese population.
Related JoVE Video
Recent advances in curdlan biosynthesis, biotechnological production, and applications.
Appl. Microbiol. Biotechnol.
PUBLISHED: 10-01-2011
Show Abstract
Hide Abstract
Curdlan is a water-insoluble ?-(1,3)-glucan produced by Agrobacterium species under nitrogen-limited condition. Its heat-induced gelling properties render curdlan to be very useful in the food industry initially. Recent advances in the understanding of the role curdlan plays in both innate and adaptive immunity lead to its growing applications in biomedicine. Our review focuses on the recent advances on curdlan biosynthesis and the improvements of curdlan fermentation production both from our laboratory and many others as well as the latest advances on the new applications of curdlan and its derivatives particularly in their immunological functions in biomedicine.
Related JoVE Video
[Aldehyde dehydrogenase 1 can be used as a new marker of cancer stem cells in laryngeal cancer cells in vitro].
Zhonghua Zhong Liu Za Zhi
PUBLISHED: 08-17-2011
Show Abstract
Hide Abstract
To detect the expression of aldehyde dehydrogenase 1 in human laryngeal cancer cells in vitro, and to explore whether it can be used as a marker of stem cells in human laryngeal cancer.
Related JoVE Video
Developmental changes and subcellular location in inhibitor of DNA binding 2 (Id2) immunoreactivity in the rat Corpus callosum.
Acta Histochem.
PUBLISHED: 08-08-2011
Show Abstract
Hide Abstract
The mechanisms underlying oligodendrocyte differentiation and myelination are still unclear, but understanding them will be critical for the development of therapies for multiple sclerosis. Inhibitor of DNA binding 2 (Id2) is a transcription factor thought to inhibit oligodendrocyte differentiation, however, it is not known whether the developmental changes and subcellular localization of Id2 are related to myelination. Therefore, we investigated the developmental changes in and the subcellular localization of Id2 immunoreactivity in the rat Corpus callosum, at post-natal developmental stages P0, P7, P14, P21, P42 and P90, by immunohistochemistry. Id2 expression increased from P0 to a peak at P42, the late stage of myelination in the Corpus callosum. Id2 immunostaining decreased slightly, but still remained high at P90. Subcellular localization of Id2 changed from presence in cytoplasm at P14 to the nuclei at P42. Moreover, Id2 was mainly co-localized with CC-1-immunopositive mature oligodendrocytes at P42. These results may be consistent with Id2 inhibitory function in oligodendrocyte differentiation, at the end of myelination or in compaction of myelin in the Corpus callosum of postnatal rat brain.
Related JoVE Video
Identification and analysis of the metabolic functions of a high-salt-tolerant halophilic aromatic yeast Candida etchellsii for soy sauce production.
World J. Microbiol. Biotechnol.
PUBLISHED: 07-13-2011
Show Abstract
Hide Abstract
Salt-tolerant yeasts are very important for the flavor formation in soy sauce fermentation production. A halophilic aromatic yeast was isolated on the basis of the molecular biological and metabolic functions from soy sauce. The ITS nucleotide sequence alignment method was used to identify the strain as Candida etchellsii by subjecting the sequence to NCBI-BLAST in comparison with that of the C. etchellsii strain Miso 0208 (a typical high-salt-tolerant halophilic aromatic yeast strain). Organic acids, amino acids and volatile flavor compounds were produced by the yeast strain which were analyzed by HPLC and SPME-GC/MS methods. Tartaric acid (0.979 ± 0.040 g/l), formic acid (0.636 ± 0.030 g/l), lactic acid (2.80 ± 0.10 g/l), ?-alkone glutaric acid (0.132 ± 0.015 g/l), citric acid (2.59 ± 0.10 g/l) and succinic acid (3.03 ± 0.20 g/l) were detected at 72 h of fermentation, respectively. Free and acid hydrolyzed amino acids at levels of 3.7355 ± 0.0027 and 11.5604 ± 0.0037 g/l, respectively, 4-ethyl guaiacols as well as other volatile flavor compounds were also detected.
Related JoVE Video
Improvement of specific growth rate of Pichia pastoris for effective porcine interferon-? production with an on-line model-based glycerol feeding strategy.
Appl. Microbiol. Biotechnol.
PUBLISHED: 07-13-2011
Show Abstract
Hide Abstract
Effective expression of porcine interferon-? (pIFN-?) with recombinant Pichia pastoris was conducted in a bench-scale fermentor. The influence of the glycerol feeding strategy on the specific growth rate and protein production was investigated. The traditional DO-stat feeding strategy led to very low cell growth rate resulting in low dry cell weight (DCW) of about 90 g/L during the subsequent induction phase. The previously reported Artificial Neural Network Pattern Recognition (ANNPR) model-based glycerol feeding strategy improved the cell density to 120 g DCW/L, while the specific growth rate decreased from 0.15 to 0.18 to 0.03-0.08 h(-1) during the last 10 h of the glycerol feeding stage leading to a variation of the porcine interferon-? production, as the glycerol feeding scheme had a significant effect on the induction phase. This problem was resolved by an improved ANNPR model-based feeding strategy to maintain the specific growth rate above 0.11 h(-1). With this feeding strategy, the pIFN-? concentration reached a level of 1.43 g/L, more than 1.5-fold higher than that obtained with the previously adopted feeding strategy. Our results showed that increasing the specific growth rate favored the target protein production and the glycerol feeding methods directly influenced the induction stage. Consequently, higher cell density and specific growth rate as well as effective porcine interferon-? production have been achieved by our novel glycerol feeding strategy.
Related JoVE Video
[Effects of andrographolide on the concentration of cytokines in BALF and the expressions of type I and III collagen mRNA in lung tissue in bleomycin-induced rat pulmonary fibrosis].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
PUBLISHED: 07-05-2011
Show Abstract
Hide Abstract
To investigate the effects of andrographolide on the concentration of TNF-? and TGF-?1 in bronchoalveolar lavage fluid (BALF) and the expressions of type I and III collagen mRNA in Lung tissue in bleomycin (BLM)-induced pulmonary fibrosis in rats.
Related JoVE Video
The reciprocal relations between teachers perceptions of childrens behavior problems and teacher-child relationships in the first preschool year.
J Genet Psychol
PUBLISHED: 06-17-2011
Show Abstract
Hide Abstract
To examine the reciprocal relations between teacher-child relationships and childrens behavior problems, the authors analyzed cross-lagged longitudinal data on teacher-child relationships and childrens internalizing and externalizing problems using a structural equation modeling approach. The homeroom teachers of 105 first-year preschoolers aged 2-3 years filled in the Student-Teacher Relationship Scale and the Child Behavior Checklist/2-3, first at 3 months after the childrens preschool entrance and then at the end of the first preschool year. Results showed significant cross-wave reciprocal relations between externalizing problems and teacher-child conflict and significant cross-wave relation from early internalizing problems to later teacher-child conflict. However, the cross-wave associations between internalizing and externalizing problems and teacher-child closeness were not significant.
Related JoVE Video
Improved curdlan fermentation process based on optimization of dissolved oxygen combined with pH control and metabolic characterization of Agrobacterium sp. ATCC 31749.
Appl. Microbiol. Biotechnol.
PUBLISHED: 05-10-2011
Show Abstract
Hide Abstract
A significant problem in scale-down cultures, rarely studied for metabolic characterization and curdlan-producing Agrobacterium sp. ATCC 31749, is the presence of dissolved oxygen (DO) gradients combined with pH control. Constant DO, between 5% and 75%, was maintained during batch fermentations by manipulating the agitation with PID system. Fermentation, metabolic and kinetic characterization studies were conducted in a scale-down system. The curdlan yield, intracellular nucleotide levels and glucose conversion efficiency into curdlan were significantly affected by DO concentrations. The optimum DO concentrations for curdlan production were 45-60%. The average curdlan yield, curdlan productivity and glucose conversion efficiency into curdlan were enhanced by 80%, 66% and 32%, respectively, compared to that at 15% DO. No apparent difference in the gel strength of the resulting curdlan was detected. The comparison of curdlan biosynthesis and cellular nucleotide levels showed that curdlan production had positive relationship with intracellular levels of UTP, ADP, AMP, NAD(+), NADH and UDP-glucose. The curdlan productivity under 45% DO and 60% DO was different during 20-50 h. However, after 60 h curdlan productivity of both conditions was similar. On that basis, a simple and reproducible two-stage DO control process for curdlan production was developed. Curdlan production yield reached 42.8 g/l, an increase of 30% compared to that of the single agitation speed control process.
Related JoVE Video
Changes of curdlan biosynthesis and nitrogenous compounds utilization characterized in ntrC mutant of Agrobacterium sp. ATCC 31749.
Curr. Microbiol.
PUBLISHED: 04-18-2011
Show Abstract
Hide Abstract
The regulatory function of global regulator NtrC on curdlan biosynthesis and nitrogen consumption under nitrogen-limited condition in Agrobacterium sp. ATCC 31749 was investigated. The ntrC mutant of Agrobacterium sp. was constructed by homologous recombination. The ability to utilize NH4Cl and KNO3 was impaired in the mutant. Other nitrogenous compounds, such as glutamic acid and glutamine, were utilized normally. Curdlan production capability was impaired severely in the mutant. Curdlan production was 5-fold lower than the wild type strain in batch fermentation with NH4Cl as the sole nitrogen source. However, up to 6.5 g l(-1) of a newly found alkali-insoluble biopolymer was produced by the ntrC mutant when glutamic acid was used as nitrogen source. The new biopolymer had glycosidic bond and hydroxyl group but no ?-configuration absorption peak on IR spectrum was found as different from curdlan. In addition, the mutant exhibited a rapid morphological change from the dot to rod form. These results deduced that the global regulator NtrC was involved in curdlan and other biopolymer biosynthesis in Agrobacterium sp. ATCC 31749 in response to nitrogen-limited condition.
Related JoVE Video
[Progress on genetics of rice grain shape trait and its related gene mapping and cloning].
Yi Chuan
PUBLISHED: 04-13-2011
Show Abstract
Hide Abstract
Increase of crop production is the primary goal of crop breeding. Rice grain shape is a quantitative trait that is directly related to yield traits and has a close relationship with quality traits. The evaluation of grain shape is mainly grain length, grain width, grain thickness, length/width, and length/thickness. In recent years, the quantitative genetic research on rice grain shape has made a significant progress and a number of important genes associated with rice grain shape have been cloned. This paper reviews the classic genetic analysis on rice grain traits, QTL mapping, grain shape gene cloning and functional analysis, and their application in rice breeding for super high yield.
Related JoVE Video
Recent advances of chitosan nanoparticles as drug carriers.
Int J Nanomedicine
PUBLISHED: 04-08-2011
Show Abstract
Hide Abstract
Chitosan nanoparticles are good drug carriers because of their good biocompatibility and biodegradability, and can be readily modified. As a new drug delivery system, they have attracted increasing attention for their wide applications in, for example, loading protein drugs, gene drugs, and anticancer chemical drugs, and via various routes of administration including oral, nasal, intravenous, and ocular. This paper reviews published research on chitosan nanoparticles, including its preparation methods, characteristics, modification, in vivo metabolic processes, and applications.
Related JoVE Video
Administration of imatinib in the first 90 days after allogeneic hematopoietic cell transplantation in patients with Philadelphia chromosome-positive acute lymphoblastic leukemia.
Chin. Med. J.
PUBLISHED: 03-03-2011
Show Abstract
Hide Abstract
Relapse happens frequently after allogeneic hematopoietic cell transplantation (allo-HCT) in the patients with Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+) ALL). Detection of the minimal residual disease (MRD) before and after allo-HCT is associated with higher relapse rate. Early administration of imatinib after allo-HCT may prevent recurrent Ph(+) ALL. The aim of this study was to evaluate the safety and efficacy of imatinib in preventing hematological relapse when imatinib was administrated in the first 90 days after allo-HCT.
Related JoVE Video
The antidepressant citalopram inhibits delayed rectifier outward K? current in mouse cortical neurons.
J. Neurosci. Res.
PUBLISHED: 02-16-2011
Show Abstract
Hide Abstract
Citalopram, a selective serotonin (5-HT) reuptake inhibitor (SSRI) as well as an antidepressant, is thought to exert its effects by increasing synaptic 5-HT levels. However, few studies have addressed the possibility that citalopram has other molecular mechanisms of action. We examined the effects of citalopram on delayed rectifier outward K(+) current (I(K) ) in mouse cortical neurons. Extracellular citalopram reversibly inhibited I(K) in a dose-dependent manner and significantly shifted both steady-state activation and inactivation curves toward hyperpolarization. Neither 5-HT itself nor antagonists of 5-HT and dopamine receptors could abolish citalopram-induced inhibition of I(K) . In addition, intracellular application of GTP?-S similarly failed to prevent the inhibition of I(K) by citalopram. When applied intracellularly, citalopram had no effect on I(K) and did not influence the reduction of I(K) induced by extracellular citalopram. The effect of citalopram was use dependent, but not frequency dependent, and it did not require channel opening. Electrophysiological recordings in acute cortical slice showed that citalopram significantly reduced the action potential (AP) firing frequency of cortical neurons and increased action potential duration (APD). The selective Kv2.1 subunit blocker Jingzhaotoxin-III (JZTX-III) did not abolish citalopram-induced I(K) inhibition. Transfection of HEK293 cells with Kv2.1 or Kv2.2 constructs indicated that citalopram mainly inhibited Kv2.2 current. We suggest that citalopram-induced inhibition of I(K) in mouse cortical neurons is independent of G-protein-coupled receptors and might exert its antidepressant effects by enhancing presynaptic efficiency. Our results may help to explain some of the unknown therapeutic effects of citalopram.
Related JoVE Video
Protective effects of catalpol on oligodendrocyte death and myelin breakdown in a rat model of chronic cerebral hypoperfusion.
Neurosci. Lett.
PUBLISHED: 02-14-2011
Show Abstract
Hide Abstract
Chronic cerebral hypoperfusion is thought to induce white matter lesions (WMLs) with oligodendrocyte (OLG) death and myelin breakdown. Although apoptosis is believed to be involved in the pathologic process of WMLs, effective therapies for such remain lacking. In the present study, we investigated whether catalpol, an iridoid glycoside, could act on oligodendrocytes (OLGs) and myelin sheaths in a rat chronic hypoperfusion model, and whether transcription factor cAMP-responsive element binding protein (CREB) phosphorylation is involved in the resulting neuroprotection. A rat model of chronic cerebral hypoperfusion was prepared by bilateral common carotid artery ligation. On the 30th day after hypoperfusion, OLG loss and myelin disruption in the ischemic white matter were more severe and evident than in the sham control. Spatial memory was also more seriously impaired in rats after hypoperfusion. Treatment with catalpol significantly suppressed diminished OLGs and myelin breakdown, and promoted the recovery of cognitive decline. The expression of Bcl-2 and phosphorylated CREB (p-CREB) was also significantly increased by catalpol treatment. In conclusion, catalpol could protect against hypoperfusion-induced WMLs and cognitive impairment through the p-CREB signaling pathway leading to downstream upregulation of Bcl-2. Our results suggest that catalpol may be a useful approach for treating cerebrovascular WMLs.
Related JoVE Video
Sequence and transcriptional analysis of the genes responsible for curdlan biosynthesis in Agrobacterium sp. ATCC 31749 under simulated dissolved oxygen gradients conditions.
Appl. Microbiol. Biotechnol.
PUBLISHED: 01-22-2011
Show Abstract
Hide Abstract
Expression at the mRNA level of ten selected genes in Agrobacterium sp. ATCC 31749 under various dissolved oxygen (DO) levels during curdlan fermentation related to electron transfer chain (ETC), tricarboxylic acid (TCA) cycle, peptidoglycan/lipopolysaccharide biosynthesis, and uridine diphosphate (UDP)-glucose biosynthesis were determined by qRT-PCR. Experiments were performed at DO levels of 30%, 50%, and 75%, as well as under low-oxygen conditions. The effect of high cell density on transcriptional response of the above genes under low oxygen was also studied. Besides cytochrome d (cyd A), the transcription levels of all the other genes were increased at higher DO and reached maximum at 50% DO. Under 75% DO, the transcriptional levels of all the genes were repressed. In addition, transcription levels of icd, sdh, cyo A, and fix N genes did not exhibit significant fluctuation with high cell density culture under low oxygen. These results suggested a mechanism for DO regulation of curdlan synthesis through regulation of transcriptional levels of ETCs, TCA, and UDP-glucose synthesis genes during curdlan fermentation. To our knowledge, this is the first report that DO concentration apparently regulates curdlan biosynthesis in Agrobacterium sp. ATCC 31749 providing essential lead for the optimization of the fermentation at the industrial scale.
Related JoVE Video
Impaired PI3K/Akt signal pathway and hepatocellular injury in high-fat fed rats.
World J. Gastroenterol.
PUBLISHED: 12-25-2010
Show Abstract
Hide Abstract
to determine whether mitochondrial dysfunction resulting from high-fat diet is related to impairment of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt, also known as PKB) pathway.
Related JoVE Video
CREB is a regulatory target for the protein kinase Akt/PKB in the differentiation of pancreatic ductal cells into islet ?-cells mediated by hepatocyte growth factor.
Biochem. Biophys. Res. Commun.
PUBLISHED: 12-04-2010
Show Abstract
Hide Abstract
We have previously reported that the PI3K/Akt signaling pathway is involved in hepatocyte growth factor (HGF)-induced differentiation of adult rat pancreatic ductal epithelial cells (PDECs) into islet ?-cells in vitro. The transcription factor CREB is one of the downstream key effectors of the PI3K/Akt signaling pathway. Recent studies showing that CREB is required for the survival of certain cell types prompted us to examine whether CREB is a nuclear target for activation via the HGF-dependent Ser/Thr kinase Akt/PKB in the differentiation of pancreatic duct cell into islet ?-cells. In this study, we first attempted to examine whether HGF modulates the Akt-dependent activation of target gene CREB and then investigated whether CREB activity affects the differentiation of HGF-induced PDECs. Finally, we studied the role of CREB in modulating the expression of transcription factors in PDECs during the differentiation of HGF-induced PDECs. Our results demonstrated that CREB is a regulatory target for the protein kinase Akt/PKB in the differentiation of pancreatic ductal cells into islet ?-cells mediated by HGF.
Related JoVE Video
[Strong association between the NINJ2 gene polymorphism and the susceptibility of stroke in Chinese Han population in Fangshan district].
Beijing Da Xue Xue Bao
PUBLISHED: 10-20-2010
Show Abstract
Hide Abstract
To investigate the association between the NINJ2 gene rs11833579 polymorphism and stroke in Han Chinese population.
Related JoVE Video
Crosstalk of the EphA2 receptor with a serine/threonine phosphatase suppresses the Akt-mTORC1 pathway in cancer cells.
Cell. Signal.
PUBLISHED: 07-16-2010
Show Abstract
Hide Abstract
Receptor tyrosine kinases of the Eph family play multiple roles in the physiological regulation of tissue homeostasis and in the pathogenesis of various diseases, including cancer. The EphA2 receptor is highly expressed in most cancer cell types, where it has disparate activities that are not well understood. It has been reported that interplay of EphA2 with oncogenic signaling pathways promotes cancer cell malignancy independently of ephrin ligand binding and receptor kinase activity. In contrast, stimulation of EphA2 signaling with ephrin-A ligands can suppress malignancy by inhibiting the Ras-MAP kinase pathway, integrin-mediated adhesion, and epithelial to mesenchymal transition. Here we show that ephrin-A1 ligand-dependent activation of EphA2 decreases the growth of PC3 prostate cancer cells and profoundly inhibits the Akt-mTORC1 pathway, which is hyperactivated due to loss of the PTEN tumor suppressor. Our results do not implicate changes in the activity of Akt upstream regulators (such as Ras family GTPases, PI3 kinase, integrins, or the Ship2 lipid phosphatase) in the observed loss of Akt T308 and S473 phosphorylation downstream of EphA2. Indeed, EphA2 can inhibit Akt phosphorylation induced by oncogenic mutations of not only PTEN but also PI3 kinase. Furthermore, it can decrease the hyperphosphorylation induced by constitutive membrane-targeting of Akt. Our data suggest a novel signaling mechanism whereby EphA2 inactivates the Akt-mTORC1 oncogenic pathway through Akt dephosphorylation mediated by a serine/threonine phosphatase. Ephrin-A1-induced Akt dephosphorylation was observed not only in PC3 prostate cancer cells but also in other cancer cell types. Thus, activation of EphA2 signaling represents a possible new avenue for anti-cancer therapies that exploit the remarkable ability of this receptor to counteract multiple oncogenic signaling pathways.
Related JoVE Video
[Coagulation and adsorption on treating the Yellow River and the impact on chlorine decay during chlorination process].
Huan Jing Ke Xue
PUBLISHED: 07-14-2010
Show Abstract
Hide Abstract
Two types of inorganic polymer coagulants, polyferric chloride (PFC) and polyaluminum chloride (PAC), were chosen to treat the Yellow River water. Different dosages were investigated in order to investigate the turbidity, UV24, DOC and permanganate index removal efficiency and their coagulation mechanisms based on the Zeta potentials. The natural organic matter removal by the combination of coagulation and adsorption with powder activated carbon were analyzed based on different coagulant and adsorbent dosages and dosing orders. The effects of combination of coagulation and adsorption on the residual chlorine decay were analyzed. The results showed that the two coagulants had high turbidity removal efficiency ( > 90%). The UV254, DOC, permanganate index removal efficiency were 29.2%, 26.1% and 27.9% respectively for PAC coagulation and were 32.3%, 23.3% and 32.9% respectively for PFC. Electric neutralization played an important role in the PAC coagulation process while both adsorption bridging and electric neutralization performed when PFC was used. The removal percentage of organic matter increased with the increase coagulant and adsorbent. The adsorption after coagulation process gave the better UV254 and DOC removal efficiency than the coagulation after adsorption. The UV254 and DOC removal efficiency were 95.2% and 99.9% for PAC coagulation after adsorption and were 90.1% and 99.9% for PFC coagulation first. But adding powder activated carbon can improve floc settlement performance and maintained persistent disinfection effect.
Related JoVE Video
Mechanisms of hepatocyte growth factor-mediated signaling in differentiation of pancreatic ductal epithelial cells into insulin-producing cells.
Biochem. Biophys. Res. Commun.
PUBLISHED: 06-11-2010
Show Abstract
Hide Abstract
Pancreatic ductal epithelial cells (PDECs) were induced to differentiate into insulin-producing cells by hepatocyte growth factor (HGF) in our previous study, but the mechanism through which this induction occurs is still unknown. HGF is a ligand that activates a tyrosine kinase encoded by the c-Met proto-oncogene. This activation is followed by indirect activation of multiple downstream signal transduction pathways (including MAPKs and the PI3K/AKT signaling pathways) that initiate various biological effects. Therefore, we speculated that the differentiation of PDECs is through either the MAPK signaling pathway or the PI3K/AKT signaling pathway. To test this hypothesis, isolated PDECs from adult rats were stimulated by adding HGF to their medium for 28days. Then, the expression levels of several protein kinases, including MAPKs (ERK1/2, p38, and JNK) and AKT, were determined by Western blotting to determine if specific protein kinases are activated in these pathways. Subsequently, re-isolated from adult rats and cultured PDECs were pre-treated with specific inhibitors of proteins shown to be activated in these signaling pathways; these cells were then induced to differentiate by the addition of HGF. The expression levels of protein kinases were determined by Western blotting, and the differentiation rate of insulin-positive cells was determined by flow cytometry. The change of PDEC differentiation rates were compared between the groups in which cells with or without inhibitors pretreatment to determine the specific signaling pathway(s) that may be involved in HGF-induced differentiation of PDECs. After isolating PDECs and stimulating them with HGF for 28days, the expression levels of phosphorylated ERK1/2 as well as total and phosphorylated AKT of cultured cells were significantly increased compared to the normal control group (p<0.05), suggesting that the signaling pathways involving ERK1/2 and Akt (MEK-ERK and PI3K-AKT) are activated during HGF-induced PDEC differentiation. MEK1/2 or PI3K inhibitors were separately added to the culture medium of PDECs pre-treated with HGF. These results show that compared to the HGF-treated group, the differentiation rate of insulin-positive cells was significantly decreased in the HGF/LY294002 (PI3K inhibitor) group (13.47+/-1.57% vs. 33.47+/-1.34%, p<0.05); however, the differentiation rate of insulin-positive cells was not significantly different in the HGF/PD98059 (MEK1/2 inhibitor) group. These data suggest that HGF induces PDECs to differentiate into insulin-producing cells through the PI3K/AKT signaling pathway.
Related JoVE Video
Improvement of the CuZn-superoxide dismutase enzyme activity and stability as a therapeutic agent by modification with polysialic acids.
Biotechnol. Lett.
PUBLISHED: 06-10-2010
Show Abstract
Hide Abstract
The optimal process for the polysialylation reaction was as follows: polysialicacid (PSA) was activated by periodate oxidation, then coupled to CuZn superoxide dismutase (SOD) with a PSA:SOD molar ratio of 40:1 for 24 h. The resulting polysialylated protein contained 3.9 ± 0.3 mol PSA per mol SOD. SDS-PAGE and atomic force microscopy revealed that the molecular weight of polysialylated SOD was about 90-100 kDa. The average size was 10-15 nm, about four-fold of the native enzyme. Compared to the native enzyme, the activity and stability of the polysialylated SOD, as well as resistance to heat, acid, alkali and proteases present in human digestive system such as pepsin and trypsin, were improved significantly as therapeutic agent.
Related JoVE Video
Removal natural organic matter by coagulation-adsorption and evaluating the serial effect through a chlorine decay model.
J. Hazard. Mater.
PUBLISHED: 06-04-2010
Show Abstract
Hide Abstract
The removal of natural organic matter (NOM) by coagulation and adsorption on modified wheat straw (MWS) was investigated. Two types of inorganic polymer coagulants, polyferric chloride (PFC) and polyaluminum chloride (PAC), were used during experiments. The removal efficiency of NOM in terms of UV(254), DOC and COD(Mn) increased with the dosage of coagulants and adsorbent increasing. Combined coagulation and adsorption showed better UV(254) and turbidity removal efficiency (61.8% and 95.8% respectively for PFC-MWS, and 61.5% and 94.2% respectively for PAC-MWS) than individual treatment. The effects of combination sequences on DOC fractionation and residual chlorine decay were analyzed and the chlorine data were fitted using a chlorine decay model. In general, the higher the molecular weight of NOM was, the better removal efficiency the combined treatment can achieve. MWS can enhance the removal of NOM with higher molecular weight. The disinfection by-products (DBPs) formation potential in the effluents from coagulation and subsequent adsorption was greater than that in the effluents from adsorption and subsequent coagulation.
Related JoVE Video
Proper distance metrics for phylogenetic analysis using complete genomes without sequence alignment.
Int J Mol Sci
PUBLISHED: 02-04-2010
Show Abstract
Hide Abstract
A shortcoming of most correlation distance methods based on the composition vectors without alignment developed for phylogenetic analysis using complete genomes is that the "distances" are not proper distance metrics in the strict mathematical sense. In this paper we propose two new correlation-related distance metrics to replace the old one in our dynamical language approach. Four genome datasets are employed to evaluate the effects of this replacement from a biological point of view. We find that the two proper distance metrics yield trees with the same or similar topologies as/to those using the old "distance" and agree with the tree of life based on 16S rRNA in a majority of the basic branches. Hence the two proper correlation-related distance metrics proposed here improve our dynamical language approach for phylogenetic analysis.
Related JoVE Video
Two-step scheme for rapid identification and differentiation of Legionella pneumophila and non-Legionella pneumophila species.
J. Clin. Microbiol.
PUBLISHED: 12-09-2009
Show Abstract
Hide Abstract
A rapid two-step scheme based on PCR amplification and enzymatic digestion analysis of a 226-bp fragment of the 16S rRNA gene was developed to identify the Legionella genus by PCR amplification and to differentiate the Legionella pneumophila and non-Legionella pneumophila species by enzymatic digestion analysis. Among 42 ATCC strains (16 strains of L. pneumophila and 26 strains of non-L. pneumophila) and 200 Legionella isolates from environmental water samples, including pools, rivers, lakes, and cooling towers in Guangdong province, 99.59% of L. pneumophila and non-L. pneumophila strains were correctly identified and differentiated by this scheme. The procedure of this two-step identification and differentiation scheme is simple and takes only about 4 h. These results suggest that this two-step scheme provides a simple and convenient method for the rapid identification and differentiation of L. pneumophila and non-L. pneumophila species.
Related JoVE Video
Amoxapine inhibits the delayed rectifier outward K+ current in mouse cortical neurons via cAMP/protein kinase A pathways.
J. Pharmacol. Exp. Ther.
PUBLISHED: 11-13-2009
Show Abstract
Hide Abstract
Ion channels are known to be modulated by antidepressant drugs, but the molecular mechanisms are not known. We have shown that the antidepressant drug amoxapine suppresses rectifier outward K(+) (I(K)) currents in mouse cortical neurons. At a concentration of 10 to 500 muM, amoxapine reversibly inhibited I(K) in a dose-dependent manner and modulated both steady-state activation and inactivation properties. The application of forskolin or dibutyryl cAMP mimicked the inhibitory effect of amoxapine on I(K) and abolished further inhibition by amoxapine. N-[2-(p-Bromocinnamylamino)ethyl]-5-iso-quinolinesulphonamide (H-89), a protein kinase A (PKA) inhibitor, augmented I(K) amplitudes and completely eliminated amoxapine inhibition of I(K). Amoxapine was also found to significantly increase intracellular cAMP levels. The effects of amoxapine on I(K) were abolished by preincubation with 5-hydroxytryptamine (5-HT) and the antagonists of 5-HT(2) receptor. Moreover, intracellular application of guanosine 5-[gammathio]-triphosphate increased I(K) amplitudes and prevented amoxapine-induced inhibition. The selective Kv2.1 subunit blocker Jingzhaotoxin-III reduced I(K) amplitudes by 30% and also significantly abolished the inhibitory effect of amoxapine. Together these results suggest that amoxapine inhibits I(K) in mouse cortical neurons by cAMP/PKA-dependent pathway associated with the 5-HT receptor, and suggest that the Kv2.1 alpha-subunit may be the target for this inhibition.
Related JoVE Video
Diagnosis, treatment and follow-up of 25 patients with melamine-induced kidney stones complicated by acute obstructive renal failure in Beijing Childrens Hospital.
Eur. J. Pediatr.
PUBLISHED: 09-30-2009
Show Abstract
Hide Abstract
A total of 25 Chinese patients aged 6 to 36 months hospitalised at Beijing Childrens Hospital due to melamine-induced kidney stones complicated by acute obstructive renal failure in 2008 were included in a study in order to diagnose and treat these special cases more effectively. Feeding history, clinical presentation, ultrasound findings, treatments and effects were summarised. Twelve to seventeen months follow-up was reported also. Ultrasound examination showed that calculi were located at the kidney and ureters. Stones were composed of both uric acid and melamine in a molar ratio of 1.2:1 to 2.1:1. Treatments providing liquid plus alkalisation of urine proved to be effective in helping the patients pass the stones. Surgical intervention was needed in severe cases. Renal function returned to normal in all 25 patients after various durations of therapy. Sixty-eight percent of the patients expelled all of the calculi within 3 months, 90% in 6 months and 95% in 9 months, without sequelae till now. Melamine-contaminated milk formula can cause kidney stones in infants, which should be diagnosed by feeding history, clinical symptoms and ultrasound examination. Composition of the stones was not only of melamine but also uric acid. Providing liquid orally or intravenously plus alkalisation of urine proved to promote the removal of the stones. Follow-up of 12 to 17 months after discharge showed no sequelae.
Related JoVE Video
Cloning of common carp SOCS-3 gene and its expression during embryogenesis, GH-transgene and viral infection.
Fish Shellfish Immunol.
PUBLISHED: 09-08-2009
Show Abstract
Hide Abstract
As a member of a newly discovered protein family, the suppressor of cytokine signalling 3 (SOCS-3) has been shown to regulate the responses of many immune cytokines in a negative auto-regulatory manner. The full-length cDNA of common carp SOCS-3 was 1603 bp and contained a 630 bp open reading frame (ORF) coding for a protein of 209 amino acids. Carp SOCS-3 molecule was well conserved especially in the SRC homology 2 (SH2) and the SOCS box. The kinase inhibitory region (KIR) and ESS domains, upstream of the SH2 domain, were conserved in carp SOCS-3, except for a specific insertion (PHRYK) in the KIR domain at the N-terminal region. Three conserved cysteine (Cys-102, 124 and 193) residues, and one additional cysteine (Cys-168) residue, were also found in carp SOCS-3. The 2015 bp genomic DNA of carp SOCS-3 contained two exons and one intron. Phylogenetic analysis showed that carp SOCS-3 sequence grouped with other known fish SOCS-3 sequences with zebrafish SOCS-3 as the closest neighbour. RT-PCR analysis showed that carp SOCS-3 was initially expressed at 4 h pf (post-fertilization) and gradually increased up to 4 w pf during embryogenesis. By RT-qPCR analysis, carp SOCS-3 gene was predominantly detected in gill, head kidney, thymus and skin, followed by spleen and peripheral blood, lower expression level was detected in kidney, intestine, liver and muscle; the SOCS-3 transcript was significantly increased in thymus, head kidney, spleen and intestine of GH (growth hormone)-transgenic carp; after SVCV (spring viraemia of carp virus) infection, the carp SOCS-3 transcript was significantly up-regulated in gill, intestine, thymus, spleen, head kidney and kidney tissues in a time-dependent manner. These results suggest that teleost SOCS-3 may play an active role in the modulation of viral-induced innate immune response and in preventing the overaction of some cytokines with viral stimulation.
Related JoVE Video
Flocculation performance of epichlorohydrin-dimethylamine polyamine in treating dyeing wastewater.
J. Environ. Manage.
PUBLISHED: 08-17-2009
Show Abstract
Hide Abstract
Epichlorohydrin-dimethylamine polymers with different intrinsic viscosity (eta) and cationicity (tau) were synthesized. The flocculation performance and mechanism of these polymers in the removal of the reactive and disperse dyes from synthetic wastewater was investigated in terms of flocculation dynamics and color removal efficiency. The polymer flocculation efficiency was compared with that of polyaluminum chloride (PAC) and a composite flocculant based on polyaluminum chloride-epichlorohydrin-dimethylamine polyamine. The results showed that epichlorohydrin-dimethylamine polymer was effective over a pH range of 2-10 for the reactive and disperse dye removal (Reactive Brilliant Red and Disperse Yellow dyes). Epichlorohydrin-dimethylamine polymer with the highest eta and tau gave the best reactive dye removal efficiency, and its adsorption-bridging and electric neutralization ability played important roles in the flocculation process. The higher the eta viscosity of the epichlorohydrin-dimethylamine polymer, the better the flocculation performance of epichlorohydrin-dimethylamine polyamine, and stronger adsorption-bridging ability was obtained for removing the disperse dye from dyeing wastewaters. Epichlorohydrin-dimethylamine polymer achieved better decolorization performance when used together with PAC.
Related JoVE Video

What is Visualize?

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

How does it work?

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

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

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