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.
Draft Genome of Chilean Honeybee (Apis mellifera) Gut Strain Lactobacillus kunkeei MP2.
Genome Announc
PUBLISHED: 10-11-2014
Show Abstract
Hide Abstract
Here, we report the first draft genome sequence of Lactobacillus kunkeei strain MP2, isolated from a Chilean honeybee gut. The sequenced genome has a total size of 1.58 Mb distributed into 44 contigs and 1,356 protein-coding sequences.
Related JoVE Video
Dendroblatta iani (Rocha e Silva-Albuquerque, 1964) n. comb. (Ectobiidae, Blattellinae).
Zootaxa
PUBLISHED: 09-16-2014
Show Abstract
Hide Abstract
The species of the family Ectobiidae range from small to large in size, and are found in tropical forests, hidden in leaf litter. This is the largest family among the cockroaches, with about 1,740 described species (Bell et al., 2007). In the Ectobiidae, the subfamilies Pseudophyllodromiinae and Blattellinae are differentiated by the position of the hook-shaped phallomere on the right or the left side, respectively, in dorsal view. The genus Xestoblatta Hebard, 1916 was included in the subfamily Blattellinae by Rocha e Silva-Albuquerque (1964), based on a specimen from the State of Pará described as Xestoblatta iani. The structures of the genitalia, the morphological characters and the habitus of the specimen, as illustrated, support the transfer of this species from the genus Xestoblatta to Dendroblatta. 
Related JoVE Video
Optical and dielectric sensors based on antimicrobial peptides for microorganism diagnosis.
Front Microbiol
PUBLISHED: 08-20-2014
Show Abstract
Hide Abstract
Antimicrobial peptides (AMPs) are natural compounds isolated from a wide variety of organisms that include microorganisms, insects, amphibians, plants, and humans. These biomolecules are considered as part of the innate immune system and are known as natural antibiotics, presenting a broad spectrum of activities against bacteria, fungi, and/or viruses. Technological innovations have enabled AMPs to be utilized for the development of novel biodetection devices. Advances in nanotechnology, such as the synthesis of nanocomposites, nanoparticles, and nanotubes have permitted the development of nanostructured platforms with biocompatibility and greater surface areas for the immobilization of biocomponents, arising as additional tools for obtaining more efficient biosensors. Diverse AMPs have been used as biological recognition elements for obtaining biosensors with more specificity and lower detection limits, whose analytical response can be evaluated through electrochemical impedance and fluorescence spectroscopies. AMP-based biosensors have shown potential for applications such as supplementary tools for conventional diagnosis methods of microorganisms. In this review, conventional methods for microorganism diagnosis as well new strategies using AMPs for the development of impedimetric and fluorescent biosensors are highlighted. AMP-based biosensors show promise as methods for diagnosing infections and bacterial contaminations as well as applications in quality control for clinical analyses and microbiological laboratories.
Related JoVE Video
New species of Oropezella Collin (Diptera, Hybotidae, Ocydromiinae) from Brazil and Costa Rica, with comments on the relationships among species-groups.
Zootaxa
PUBLISHED: 08-18-2014
Show Abstract
Hide Abstract
The following sixteen new species of Oropezella from Brazil and Costa Rica are described and illustrated: Oropezella aquila sp. nov., O. arcuata sp. nov., O. bicolor sp. nov., O. bipunctata sp. nov., O. clavata sp. nov., O. costaricensis sp. nov., O. falcata sp. nov., O. flavida sp. nov., O. granulosa sp. nov., O. longifrons sp. nov., O. planti sp. nov., O. plaumanni sp. nov., O. rafaeli sp. nov., O. spathula sp. nov., O. uncata sp. nov. and O. undulata sp. nov. A new combination is proposed: Oropezella amazonica (Rafael & Ale-Rocha, 1990) is transferred from Ocydromia. Additionally, an identification key to all 25 Neotropical and Andean species of Oropezella is provided. A cladistic analysis of Oropezella using 29 species from the Neotropical, Andean, Palearctic and Neozelandic regions is performed and its results and implications discussed. 
Related JoVE Video
Synthetic biology approaches to improve biocatalyst identification in metagenomic library screening.
Microb Biotechnol
PUBLISHED: 08-13-2014
Show Abstract
Hide Abstract
There is a growing demand for enzymes with improved catalytic performance or tolerance to process-specific parameters, and biotechnology plays a crucial role in the development of biocatalysts for use in industry, agriculture, medicine and energy generation. Metagenomics takes advantage of the wealth of genetic and biochemical diversity present in the genomes of microorganisms found in environmental samples, and provides a set of new technologies directed towards screening for new catalytic activities from environmental samples with potential biotechnology applications. However, biased and low level of expression of heterologous proteins in Escherichia coli together with the use of non-optimal cloning vectors for the construction of metagenomic libraries generally results in an extremely low success rate for enzyme identification. The bottleneck arising from inefficient screening of enzymatic activities has been addressed from several perspectives; however, the limitations related to biased expression in heterologous hosts cannot be overcome by using a single approach, but rather requires the synergetic implementation of multiple methodologies. Here, we review some of the principal constraints regarding the discovery of new enzymes in metagenomic libraries and discuss how these might be resolved by using synthetic biology methods.
Related JoVE Video
Expanding the logic of bacterial promoters using engineered overlapping operators for global regulators.
ACS Synth Biol
PUBLISHED: 07-28-2014
Show Abstract
Hide Abstract
The understanding of how the architecture of cis-regulatory elements at bacterial promoters determines their final output is of central interest in modern biology. In this work, we attempt to gain insight into this process by analyzing complex promoter architectures in the model organism Escherichia coli. By focusing on the relationship between different TFs at the genomic scale in terms of their binding site arrangement and their effect on the target promoters, we found no strong constraint limiting the combinatorial assembly of TF pairs in E. coli. More strikingly, overlapping binding sites were found equally associated with both equivalent (both TFs have the same effect on the promoter) and opposite (one TF activates while the other repress the promoter) effects on gene expression. With this information on hand, we set an in silico approach to design overlapping sites for three global regulators (GRs) of E. coli, specifically CRP, Fis, and IHF. Using random sequence assembly and an evolutionary algorithm, we were able to identify potential overlapping operators for all TF pairs. In order to validate our prediction, we constructed two lac promoter variants containing overlapping sites for CRP and IHF designed in silico. By assaying the synthetic promoters using a GFP reporter system, we demonstrated that these variants were functional and activated by CRP and IHF in vivo. Taken together, presented results add new information on the mechanisms of signal integration in bacterial promoters and provide new strategies for the engineering of synthetic regulatory circuits in bacteria.
Related JoVE Video
Insecticidal and genotoxic potential of two semi-synthetic derivatives of dillapiole for the control of Aedes (Stegomyia) aegypti (Diptera: Culicidae).
Mutat Res Genet Toxicol Environ Mutagen
PUBLISHED: 07-21-2014
Show Abstract
Hide Abstract
The effects of two semi-synthetic dillapiole derivatives, ethyl-ether dillapiole and n-butyl ether dillapiole, on eggs and larvae of Aedes aegypti were studied in view of the need for expansion and renovation of strategic action to control this mosquito - the vector of Dengue virus -, which currently shows a high resistance to chemical insecticides. Eggs and third-instar larvae of A. aegypti that had been exposed to different concentrations of these two compounds showed toxicity and susceptibility, with 100% mortality. Classical cytogenetic assays showed genotoxicity caused by the two compounds in A. aegypti from the cumulative effect of nuclear abnormalities, indicating that these derivatives may be potential alternatives to control A. aegypti.
Related JoVE Video
Statins increase rifampin mycobactericidal effect.
Antimicrob. Agents Chemother.
PUBLISHED: 07-21-2014
Show Abstract
Hide Abstract
Mycobacterium leprae and Mycobacterium tuberculosis antimicrobial resistance has been followed with great concern during the last years, while the need for new drugs able to control leprosy and tuberculosis, mainly due to extensively drug-resistant tuberculosis (XDR-TB), is pressing. Our group recently showed that M. leprae is able to induce lipid body biogenesis and cholesterol accumulation in macrophages and Schwann cells, facilitating its viability and replication. Considering these previous results, we investigated the efficacies of two statins on the intracellular viability of mycobacteria within the macrophage, as well as the effect of atorvastatin on M. leprae infections in BALB/c mice. We observed that intracellular mycobacteria viability decreased markedly after incubation with both statins, but atorvastatin showed the best inhibitory effect when combined with rifampin. Using Shepard's model, we observed with atorvastatin an efficacy in controlling M. leprae and inflammatory infiltrate in the BALB/c footpad, in a serum cholesterol level-dependent way. We conclude that statins contribute to macrophage-bactericidal activity against Mycobacterium bovis, M. leprae, and M. tuberculosis. It is likely that the association of statins with the actual multidrug therapy effectively reduces mycobacterial viability and tissue lesion in leprosy and tuberculosis patients, although epidemiological studies are still needed for confirmation.
Related JoVE Video
Multidrug-resistant nontuberculous mycobacteria isolated from cystic fibrosis patients.
J. Clin. Microbiol.
PUBLISHED: 06-11-2014
Show Abstract
Hide Abstract
Worldwide, nontuberculous mycobacteria (NTM) have become emergent pathogens of pulmonary infections in cystic fibrosis (CF) patients, with an estimated prevalence ranging from 5 to 20%. This work investigated the presence of NTM in sputum samples of 129 CF patients (2 to 18 years old) submitted to longitudinal clinical supervision at a regional reference center in Rio de Janeiro, Brazil. From June 2009 to March 2012, 36 NTM isolates recovered from 10 (7.75%) out of 129 children were obtained. Molecular identification of NTM was performed by using PCR restriction analysis targeting the hsp65 gene (PRA-hsp65) and sequencing of the rpoB gene, and susceptibility tests were performed that followed Clinical and Laboratory Standards Institute recommendations. For evaluating the genotypic diversity, pulsed-field gel electrophoresis (PFGE) and/or enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) was performed. The species identified were Mycobacterium abscessus subsp. bolletii (n = 24), M. abscessus subsp. abscessus (n = 6), Mycobacterium fortuitum (n = 3), Mycobacterium marseillense (n = 2), and Mycobacterium timonense (n = 1). Most of the isolates presented resistance to five or more of the antimicrobials tested. Typing profiles were mainly patient specific. The PFGE profiles indicated the presence of two clonal groups for M. abscessus subsp. abscessus and five clonal groups for M. abscesssus subsp. bolletii, with just one clone detected in two patients. Given the observed multidrug resistance patterns and the possibility of transmission between patients, we suggest the implementation of continuous and routine investigation of NTM infection or colonization in CF patients, including countries with a high burden of tuberculosis disease.
Related JoVE Video
Multilocus sequence typing scheme versus pulsed-field gel electrophoresis for typing Mycobacterium abscessus isolates.
J. Clin. Microbiol.
PUBLISHED: 06-04-2014
Show Abstract
Hide Abstract
Outbreaks of infections by rapidly growing mycobacteria following invasive procedures, such as ophthalmological, laparoscopic, arthroscopic, plastic, and cardiac surgeries, mesotherapy, and vaccination, have been detected in Brazil since 1998. Members of the Mycobacterium chelonae-Mycobacterium abscessus group have caused most of these outbreaks. As part of an epidemiological investigation, the isolates were typed by pulsed-field gel electrophoresis (PFGE). In this project, we performed a large-scale comparison of PFGE profiles with the results of a recently developed multilocus sequence typing (MLST) scheme for M. abscessus. Ninety-three isolates were analyzed, with 40 M. abscessus subsp. abscessus isolates, 47 M. abscessus subsp. bolletii isolates, and six isolates with no assigned subspecies. Forty-five isolates were obtained during five outbreaks, and 48 were sporadic isolates that were not associated with outbreaks. For MLST, seven housekeeping genes (argH, cya, glpK, gnd, murC, pta, and purH) were sequenced, and each isolate was assigned a sequence type (ST) from the combination of obtained alleles. The PFGE patterns of DraI-digested DNA were compared with the MLST results. All isolates were analyzable by both methods. Isolates from monoclonal outbreaks showed unique STs and indistinguishable or very similar PFGE patterns. Thirty-three STs and 49 unique PFGE patterns were identified among the 93 isolates. The Simpson's index of diversity values for MLST and PFGE were 0.69 and 0.93, respectively, for M. abscessus subsp. abscessus and 0.96 and 0.97, respectively, for M. abscessus subsp. bolletii. In conclusion, the MLST scheme showed 100% typeability and grouped monoclonal outbreak isolates in agreement with PFGE, but it was less discriminative than PFGE for M. abscessus.
Related JoVE Video
Reuse of refinery's tertiary-treated wastewater in cooling towers: microbiological monitoring.
Environ Sci Pollut Res Int
PUBLISHED: 05-29-2014
Show Abstract
Hide Abstract
The study was planned to quantify the distribution of bacteria between bulk water and biofilm formed on different materials in an industrial scale cooling tower system of an oil refinery operating with clarified and chlorinated freshwater (CCW) or chlorinated tertiary effluent (TRW) as makeup water. The sessile and planktonic heterotrophic bacteria and Pseudomonas aeruginosa densities were significantly higher in the cooling tower supplied with clarified and chlorinated freshwater (CTCW) (p?
Related JoVE Video
Chromosomal integration of transcriptional fusions.
Methods Mol. Biol.
PUBLISHED: 05-14-2014
Show Abstract
Hide Abstract
The characterization and parameterization of promoters is crucial for the study of gene regulatory networks. While a number of techniques are available for this purpose, the use of reporter fusions integrated in the chromosome of a bacterial host affords precise quantification of transcriptional responses with high reproducibility. Here, we describe the integration of green fluorescent protein (GFP) and lacZ reporter cassettes using either mini-Tn7-based vectors or homologous chromosomal recombination to analyze gene regulation at transcriptional and post-transcriptional levels.
Related JoVE Video
The reliability of self-assessment of affective state in different phases of bipolar disorder.
J. Nerv. Ment. Dis.
PUBLISHED: 04-15-2014
Show Abstract
Hide Abstract
Some studies have indicated that the capacity of self-assessment of affective state is more compromised during mania than during depression. In the present study, we investigated whether the reliability of self-assessment in bipolar disorder varies as a function of actual affective state (i.e., euthymia, mania, or depression). Sixty-five patients with a diagnosis of type I and type II bipolar disorder were evaluated with regard to the occurrence of an affective syndrome using the Clinical Global Impressions Scale for use in bipolar illness, the Positive and Negative Syndrome Scale, and the Global Assessment of Functioning scale. In parallel, we applied the Analog Visual Mood Scale, a self-assessment tool to evaluate mood changes. The same individual prospectively completed the self-assessment scale in different affective states. During depression, the patients' evaluation was significantly different from when they were in manic or euthymic mood states. However, when in mania, the patients evaluated their mood state similarly to when they were euthymic. The bipolar patients in mania but not in depression did not reliably evaluate themselves with regard to their affective state.
Related JoVE Video
Brazil nuts are subject to infection with B and G aflatoxin-producing fungus, Aspergillus pseudonomius.
Int. J. Food Microbiol.
PUBLISHED: 03-27-2014
Show Abstract
Hide Abstract
The exploitation of the Brazil nut is one of the most important activities of the extractive communities of the Amazon rainforest. However, its commercialization can be affected by the presence of aflatoxins produced by fungi, namely Aspergillus section Flavi. In the present study, we investigated a collection of Aspergillus nomius strains isolated from Brazil nuts using different approaches, including morphological characters, RAPD and AFLP profiles, partial ?-tubulin and calmodulin nucleotide sequences, aflatoxin patterns, as well as tolerance to low water activity in cultured media. Results showed that most of the isolates do belong to A. nomius species, but a few were re-identified as Aspergillus pseudonomius, a very recently described species. The results of the analyses of molecular variance, as well as the high pairwise FST values between A. nomius and A. pseudonomius suggested the isolation between these two species and the inexistence of gene flow. Fixed interspecific nucleotide polymorphisms at ?-tubulin and calmodulin loci are presented. All A. pseudonomius strains analyzed produced aflatoxins AFB1, AFB2, AFG1 and AFG2. This study contains the first-ever report on the occurrence in Brazil nuts of A. pseudonomius. The G-type aflatoxins and the mycotoxin tenuazonic acid are reported here for the first time in A. pseudonomius.
Related JoVE Video
Phylogenetic analysis of the GST family in Anopheles (Nyssorhynchus) darlingi.
Acta Trop.
PUBLISHED: 03-24-2014
Show Abstract
Hide Abstract
Anopheles darlingi Root, 1926 and Anopheles gambiae (Diptera: Culicidae) are the most important human malaria vectors in South America and Africa, respectively. The two species are estimated to have diverged 100 million years ago. Studies on the phylogenetics and evolution of gene sequences, such as glutathione S-transferase (GST) in disease-transmitting mosquitoes are scarce. The sigma class GST (KC890767) from the transcriptome of An. darlingi captured in the Brazilian Amazon was studied by in silico hybridization, and mapped to chromosome 3 of An. gambiae. The sigma class GST of An. darlingi was used for phylogenetic analyses to understand the GST base composition of the most recent common ancestor between An. darlingi, Anopheles gambiae, Aedes aegypti and Culex quinquefasciatus. The GST (KC890767) of An. darlingi was studied to generate the main divergence branches using a Neighbor-Joining and bootstrapping approaches to confirm confidence levels on the tree nodes that separate the An. darlingi and other mosquito species. The results showed divergence between An. gambiae, Ae. Aegypti, Cx. quinquefasciatus, and Phlebotomus papatasi as outgroup, and the homology relationship between sigma class GST of An. darlingi and GSTS1_1 gene of An. gambiae was valuable for phylogenetic and evolutionary studies.
Related JoVE Video
Cobalt-embedded nitrogen-rich carbon nanotubes efficiently catalyze hydrogen evolution reaction at all pH values.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 03-20-2014
Show Abstract
Hide Abstract
Despite being technically possible, splitting water to generate hydrogen is still practically unfeasible due mainly to the lack of sustainable and efficient catalysts for the half reactions involved. Herein we report the synthesis of cobalt-embedded nitrogen-rich carbon nanotubes (NRCNTs) that 1) can efficiently electrocatalyze the hydrogen evolution reaction (HER) with activities close to that of Pt and 2) function well under acidic, neutral or basic media alike, allowing them to be coupled with the best available oxygen-evolving catalysts-which also play crucial roles in the overall water-splitting reaction. The materials are synthesized by a simple, easily scalable synthetic route involving thermal treatment of Co(2+) -embedded graphitic carbon nitride derived from inexpensive starting materials (dicyandiamide and CoCl2 ). The materials' efficient catalytic activity is mainly attributed to their nitrogen dopants and concomitant structural defects.
Related JoVE Video
[Semantic equivalence of the Brazilian Portuguese-language version of the Online Cognition Scale].
Cad Saude Publica
PUBLISHED: 03-07-2014
Show Abstract
Hide Abstract
This study evaluated the semantic equivalence of the Online Cognition Scale in Brazilian Portuguese. The process included five steps: translation, back-translation, technical review, evaluation of semantic equivalence by trained professionals, and evaluation of the instrument for content understanding by a group of professionals (n = 10) and students (n = 37). The instrument was translated and adapted to Portuguese, showing a high level of verbal comprehension by the target population. The adapted version for use in Brazil resulted in an equivalent instrument from the semantic and content point of view, ready for evaluation of its psychometric qualities in the Brazilian cultural context.
Related JoVE Video
Infliximab attenuates inflammatory osteolysis in a model of periodontitis in Wistar rats.
Exp. Biol. Med. (Maywood)
PUBLISHED: 02-28-2014
Show Abstract
Hide Abstract
Periodontitis is a chronic inflammatory disease related to tooth loss in adults. Infliximab is a chimeric monoclonal antibody against TNF-? and is prescribed for the treatment of systemic inflammatory diseases. This study aimed to investigate the role of infliximab on experimental periodontal disease (EPD). EPD was induced by passing a 3.0 nylon thread around the upper left second molar in Wistar rats. Animals were either treated with intravenous infliximab (1, 5, 7, and 10 mg/kg) or saline solution 30 min before the periodontitis induction and were followed until they were sacrificed on the 11th day. A subset of rats was euthanized on the third day for analysis of gingival myeloperoxidase (MPO) and the blood MPO granulocyte index. In addition, we analyzed the bone loss index (BLI), the periodontal histopathological score, and the periodontal collagen network using confocal microscopy. We also analyzed metalloproteinase-1/-8, RANK, RANK-L, and osteoprotegerin in maxillary tissue by immunohistochemistry Gingival MPO, IL-1?, TNF-? were measured by ELISA. EPD caused leukocytosis, significant increases in BLI and gingival pro-inflammatory cytokines and cell infiltrates, with worse histopathological scores and periodontal collagen derangement. Infliximab (5 mg/kg) reduced granulocyte blood counts, gingival IL-1?, TNF-?, and MPO levels, diminished MMP-1/-8, RANK, and RANK-L bone immunolabeling with better periodontal histopathological scores and collagen network in comparison with the challenged saline group. We concluded that infliximab had significant anti-inflammatory and bone-protective effects in Wistar rats challenged by periodontitis.
Related JoVE Video
Spontaneous closure of post-intervention left anterior descending coronary pseudoaneurysm.
Rev Port Cardiol
PUBLISHED: 02-27-2014
Show Abstract
Hide Abstract
Coronary pseudoaneurysms are an unusual finding during coronary angiography and there are very little data on their prognosis in the literature. We report the case of a 62-year-old man admitted with an anterior myocardial infarction who developed a pseudoaneurysm in the mid left anterior descending artery some days after a type I coronary perforation during coronary angioplasty. Spontaneous closure of the pseudoaneurysm was observed during hospital follow-up. Spontaneous closure of coronary pseudoaneurysms may be more common in clinical practice than previously thought, but few cases have been reported. As the natural history of post-intervention coronary pseudoaneurysms has been little investigated, reports of their occurrence may help to clarify their evolution.
Related JoVE Video
Proteomic identification of differentially expressed proteins during the acquisition of somatic embryogenesis in oil palm (Elaeis guineensis Jacq.).
J Proteomics
PUBLISHED: 02-24-2014
Show Abstract
Hide Abstract
In the present study we have identified and characterized the proteins expressed during different developmental stages of Elaeis guineensis calli obtained from zygotic embryos. We were interested in the possible proteomic changes that would occur during the acquisition of somatic embryogenesis and therefore samples were collected from zygotic embryos (E1), swollen explants 14days (E2) in induction medium, primary callus (E3), and pro-embryogenic callus (E4). The samples were grinded in liquid nitrogen, followed by total protein extraction using phenol and extraction buffer. Proteins were analyzed by two-dimensional electrophoresis (2-DE) and the differentially expressed protein spots were analyzed by MALDI-TOF mass spectrometry (MS and MS/MS). Interestingly, we have identified proteins, which can be used as potential candidates for future studies aiming at the development of biomarkers for embryogenesis acquisition and for the different stages leading to pro-embryogenic callus formation such as type IIIa membrane protein cp-wap13, fructokinase and PR proteins. The results obtained shed some light on the biochemical events involved in the process of somatic embryogenesis of E. guineensis obtained from zygotic embryos. The use of stage-specific protein markers can help monitor cell differentiation and contribute to improve the protocols for successfully cloning the species.
Related JoVE Video
The differential response of the Pben promoter of Pseudomonas putida?mt-2 to BenR and XylS prevents metabolic conflicts in m-xylene biodegradation.
Environ. Microbiol.
PUBLISHED: 02-13-2014
Show Abstract
Hide Abstract
Pseudomonas putida?mt-2 encompasses two alternative and potentially conflicting routes for benzoate metabolism, one meta pathway encoded by xyl genes of the pWW0 plasmid and mastered by the Pm promoter and XylS, and one chromosomally encoded ortho pathway initiated by Pben and the BenR protein. Any cross-activation of Pben promoter by XylS ought to cause a metabolic conflict during the degradation of m-xylene because 3-methylbenzoate (3MBz) generated as an intermediate can be channelled through the ortho pathway and produce toxic dead-end metabolites. The activation of Pben by XylS was revisited using both reporter technology and tiling arrays targeted to the sequences of interest around messenger RNA initiation of both Pben and Pm promoters. Analysis of supersensitive luxCDABE fusions, inspection of xylX versus benA transcripts and growth tests of benR mutants indicated that the natural expression ranges of XylS under various conditions are insufficient to cause a significant cross-regulation of Pben whether cells face endogenous or exogenous 3MBz. This seems to stem from the nature of the operators for binding either transcriptional factor, which in the case of the Pben promoter of P.?putida?mt-2 appear to have evolved for avoiding a strong interaction with XylS.
Related JoVE Video
Genetic and ElectroNic medIcal records to predict oUtcomeS in Heart Failure patients (GENIUS-HF) - design and rationale.
BMC Cardiovasc Disord
PUBLISHED: 02-10-2014
Show Abstract
Hide Abstract
Studies adopting electronic medical records and genomic information are becoming widespread. Through this new modality in research, it is possible to study how genetic variants influence susceptibility towards chronic conditions and can improve patient care.Our aim is to develop a biobank with 2,000 heart failure patients treated in a tertiary cardiology hospital containing electronic medical records data and biologic samples for performing genome-wide association studies for validation and development of medical decision routines aimed at helping the clinical management of patients.
Related JoVE Video
3'-nucleotidase/nuclease activity allows Leishmania parasites to escape killing by neutrophil extracellular traps.
Infect. Immun.
PUBLISHED: 02-10-2014
Show Abstract
Hide Abstract
Leishmaniasis is a widespread neglected tropical disease caused by parasites of the Leishmania genus. These parasites express the enzyme 3'-nucleotidase/nuclease (3'NT/NU), which has been described to be involved in parasite nutrition and infection. Bacteria that express nucleases escape the toxic effects of neutrophil extracellular traps (NETs). Hence, we investigated the role of 3'NT/NU in Leishmania survival of NET-mediated killing. Promastigotes of Leishmania infantum were cultured in high-phosphate (HP) or low-phosphate (LP) medium to modulate nuclease activity. We compared the survival of the two different groups of Leishmania during interaction with human neutrophils, assessing the role of neutrophil extracellular traps. As previously reported, we detected higher nuclease activity in parasites cultured in LP medium. Both LP and HP promastigotes were capable of inducing the release of neutrophil extracellular traps from human neutrophils in a dose- and time-dependent manner. LP parasites had 2.4 times more survival than HP promastigotes. NET disruption was prevented by the treatment of the parasites with ammonium tetrathiomolybdate (TTM), a 3'NT/NU inhibitor. Inhibition of 3'NT/NU by 3'-AMP, 5'-GMP, or TTM decreased promastigote survival upon interaction with neutrophils. Our results show that Leishmania infantum induces NET release and that promastigotes can escape NET-mediated killing by 3'-nucleotidase/nuclease activity, thus ascribing a new function to this enzyme.
Related JoVE Video
Subregional basal forebrain atrophy in Alzheimer's disease: a multicenter study.
J. Alzheimers Dis.
PUBLISHED: 02-08-2014
Show Abstract
Hide Abstract
Histopathological studies in Alzheimer's disease (AD) suggest severe and region-specific neurodegeneration of the basal forebrain cholinergic system (BFCS). Here, we studied the between-center reliability and diagnostic accuracy of MRI-based BFCS volumetry in a large multicenter data set, including participants with prodromal (n = 41) or clinically manifest AD (n = 134) and 148 cognitively healthy controls. Atrophy was determined using voxel-based and region-of-interest based analyses of high-dimensionally normalized MRI scans using a newly created map of the BFCS based on postmortem in cranio MRI and histology. The AD group showed significant volume reductions of all subregions of the BFCS, which were most pronounced in the posterior nucleus basalis Meynert (NbM). The mild cognitive impairment-AD group showed pronounced volume reductions in the posterior NbM, but preserved volumes of anterior-medial regions. Diagnostic accuracy of posterior NbM volume was superior to hippocampus volume in both groups, despite higher multicenter variability of the BFCS measurements. The data of our study suggest that BFCS morphometry may provide an emerging biomarker in AD.
Related JoVE Video
Selection of candidate reference genes for real-time PCR studies in lettuce under abiotic stresses.
Planta
PUBLISHED: 01-31-2014
Show Abstract
Hide Abstract
The process of selection and validation of reference genes is the first step in studies of gene expression by real-time quantitative polymerase chain reaction (RT-qPCR). The genome of lettuce, the most popular leaf vegetable cultivated worldwide, has recently been sequenced; therefore, suitable reference genes for reliable results in RT-qPCR analyses are required. In the present study, 17 candidate reference genes were selected, and their expression stability in lettuce leaves under drought, salt, heavy metal, and UV-C irradiation conditions and under the application of abscisic acid (ABA) was evaluated using geNorm and NormFinder software. The candidate reference genes included protein-coding traditional and novel reference genes and microRNAs (miRNAs). The results indicate that the expression stability is dependent on the experimental conditions. The novel protein-coding reference genes were more suitable than the traditional reference genes under drought, UV-C irradiation, and heavy metal conditions and under the application of ABA. Only under salinity conditions were the traditional protein-coding reference genes more stable than the novel genes. In addition, the miRNAs, mainly MIR169, MIR171/170 and MIR172, were stably expressed under the abiotic stresses evaluated, representing a suitable alternative approach for gene expression data normalization. The expression of phenylalanine ammonia lyase (PAL) and 4-hydroxyphenylpyruvate dioxygenase (HPPD) was used to further confirm the validated protein-coding reference genes, and the expression of MIR172 and MIR398 was used to confirm the validated miRNA genes, showing that the use of an inappropriate reference gene induces erroneous results. This work is the first survey of the stability of reference genes in lettuce and provides guidelines to obtain more accurate RT-qPCR results in lettuce studies.
Related JoVE Video
Carotenoid biosynthetic and catabolic pathways: gene expression and carotenoid content in grains of maize landraces.
Nutrients
PUBLISHED: 01-31-2014
Show Abstract
Hide Abstract
Plant carotenoids have been implicated in preventing several age-related diseases, and they also provide vitamin A precursors; therefore, increasing the content of carotenoids in maize grains is of great interest. It is not well understood, however, how the carotenoid biosynthetic pathway is regulated. Fortunately, the maize germplasm exhibits a high degree of genetic diversity that can be exploited for this purpose. Here, the accumulation of carotenoids and the expression of genes from carotenoid metabolic and catabolic pathways were investigated in several maize landraces. The carotenoid content in grains varied from 10.03, in the white variety MC5, to 61.50 ?g·g?¹, in the yellow-to-orange variety MC3, and the major carotenoids detected were lutein and zeaxanthin. PSY1 (phythoene synthase) expression showed a positive correlation with the total carotenoid content. Additionally, the PSY1 and HYD3 (ferredoxin-dependent di-iron monooxygenase) expression levels were positively correlated with ?-cryptoxanthin and zeaxanthin, while CYP97C (cytochrome P450-type monooxygenase) expression did not correlate with any of the carotenoids. In contrast, ZmCCD1 (carotenoid dioxygenase) was more highly expressed at the beginning of grain development, as well as in the white variety, and its expression was inversely correlated with the accumulation of several carotenoids, suggesting that CCD1 is also an important enzyme to be considered when attempting to improve the carotenoid content in maize. The MC27 and MC1 varieties showed the highest HYD3/CYP97C ratios, suggesting that they are promising candidates for increasing the zeaxanthin content; in contrast, MC14 and MC7 showed low HYD3/CYP97C, suggesting that they may be useful in biofortification efforts aimed at promoting the accumulation of provitamin A. The results of this study demonstrate the use of maize germplasm to provide insight into the regulation of genes involved in the carotenoid pathway, which would thus better enable us to select promising varieties for biofortification efforts.
Related JoVE Video
[Styles of nurses' caring in Intensive Therapy mediated by technology].
Rev Bras Enferm
PUBLISHED: 01-21-2014
Show Abstract
Hide Abstract
The objective was to identify the social representations of the intensive therapy nurses about the care practices in face of the technology. Participant observation and interviews were conducted with twenty one nurses of an intensive therapy center, at a Rio de Janeiro public hospital. Lexical analysis was applied, using the Alceste 2010. The results were organized in two categories, each of them with three lexical classes. The first brought the meaning of the technology in intensive therapy and the organization of the styles of caring; the second showed the client's condition and its impact on nurses' care practices. It was concluded that the technology organizes the nurses' styles of caring, that are built on client assistance in the work context. These lead nurses to assign meaning to their practices driving them to the elaboration of ways of acting in face of the technologies.
Related JoVE Video
Comparative metabolism of cellulose, sophorose and glucose in Trichoderma reesei using high-throughput genomic and proteomic analyses.
Biotechnol Biofuels
PUBLISHED: 01-14-2014
Show Abstract
Hide Abstract
The filamentous fungus Trichoderma reesei is a major producer of lignocellulolytic enzymes utilized by bioethanol industries. However, to achieve low cost second generation bioethanol production on an industrial scale an efficient mix of hydrolytic enzymes is required for the deconstruction of plant biomass. In this study, we investigated the molecular basis for lignocellulose-degrading enzyme production T. reesei during growth in cellulose, sophorose, and glucose.
Related JoVE Video
Isolation of high-quality RNA from grains of different maize varieties.
Prep. Biochem. Biotechnol.
PUBLISHED: 01-10-2014
Show Abstract
Hide Abstract
The study of gene expression in maize varieties represents a powerful tool aiming to increase vitamin A precursors. However, the isolation of RNA from different maize varieties is challenging because these varieties show different levels of polysaccharides, and most methods available for RNA isolation are inappropriate for grain samples. The polysaccharides co-purify and co-precipitate with RNA during isolation, resulting in low-quality RNA, compromising the use of RNA in subsequent applications. Thus, a cetyltrimethylammonium bromide (CTAB)-based method was adapted in this study and compared with six methods for RNA isolation, including commercial reagents and RNA and DNA isolation kits, in order to identify the most appropriate for maize grains from different varieties. Most of the methods evaluated were considered inadequate due to limitations in terms of purity and/or quantity of the isolated RNA, which affected the efficiency of subsequent RT-qPCR analysis, resulting in nonamplification of ?-carotene hydroxylase gene (HYD3) or high deviation among replicates. However, the CTAB modified method allowed the study to obtain intact RNA, with high quality and quantity, from 25 maize varieties. Furthermore, this RNA was successfully used to evaluate the expression of HYD3 gene by real-time qualitative polymerase chain reaction (RT-qPCR), and thus represents a simple, efficient, and low-cost strategy.
Related JoVE Video
[Technology in nursing care: an analysis from the conceptual framework of Fundamental Nursing].
Rev Bras Enferm
PUBLISHED: 01-08-2014
Show Abstract
Hide Abstract
This is a qualitative, field research, whose purpose was to discuss the use of technologies in the nursing care in intensive therapy, taking as reference the theoretical conceptual framework of Fundamental Nursing. Observation and interviews were conducted with twenty two nurses of an intensive therapy unit, with ethnographic analysis. The technology, from the domain of a technological language, provides conditions so that the fundamentals of the nursing care can be effectively incorporated to the nurse practice. The idea of dehumanization linked to the technology can be explained by the way that the nurse ads sense to the things related to his daily life, which will guide his action. The conclusion is that the technologies help to promote life and to rescue the human.
Related JoVE Video
Vitamin D and systemic lupus erythematosus: state of the art.
Clin. Rheumatol.
PUBLISHED: 01-07-2014
Show Abstract
Hide Abstract
Systemic lupus erythematosus (SLE) is a systemic inflammatory disease associated with genetic, environmental, hormonal, and immunological factors. One of these factors is vitamin D deficiency. Vitamin D plays many roles in the immune system. Several studies have suggested a potential role in the development of autoimmune diseases. SLE patients have low serum levels of vitamin D, which increase the possibility of an association between vitamin deficiency and disease onset and evolution. This review of the literature presents an analysis of the aspects related to the immunoregulatory effects of vitamin D and its importance for SLE, as well as the recommendations for vitamin D supplementation in these patients.
Related JoVE Video
Expression pattern of cellulolytic and xylanolytic genes regulated by transcriptional factors XYR1 and CRE1 are affected by carbon source in Trichoderma reesei.
Gene Expr. Patterns
PUBLISHED: 01-07-2014
Show Abstract
Hide Abstract
Trichoderma reesei is the most important fungus for the industrial production of enzymes to biomass deconstruction. Most of the genes encoding cellulases and hemicellulases are regulated by the transcription factors CRE1 and XYR1. In this work, the regulation of 22 genes of cellulases and xylanases by these transcription factors was investigated under three different carbon sources. Analysis of gene expression and enzymatic profiles of CMCase, ?-glucosidase, and xylanases showed different regulation that was depended of the carbon source in both ?xyr1 and ?cre1 mutants. In the presence of glucose, the majority of genes evaluated (82%) showed increased expression levels in the ?cre1 mutant compared to the parental QM9414 strain. In the ?xyr1 mutant, it was observed that expression of cellulase and xylanase genes was reduced compared to the parental QM9414 strain, when cultured in the presence of cellulose or sophorose. Interesting, in the presence of glucose, approximately 60% of the analyzed genes had increased expression in the ?xyr1 mutant compared to parental strain. Furthermore, no correlation between gene expression and the number of putative binding sites of XYR1 and CRE1 to promoter region of cellulolytic and xylanolytic studied genes was observed. Therefore, these results demonstrated that the regulation of cellulase and xylanase by the transcription factors CRE1 and XYR1 is influenced by different carbon sources.
Related JoVE Video
A continuous and direct assay to monitor leucine-rich repeat kinase 2 activity.
Anal. Biochem.
PUBLISHED: 01-07-2014
Show Abstract
Hide Abstract
Leucine-rich repeat kinase 2 (LRRK2) is a multi-domain enzyme displaying activities of GTP hydrolase and protein threonine/serine kinase in separate domains. Mutations in both catalytic domains have been linked to the onset of Parkinson's disease, which triggered high interest in this enzyme as a potential target for drug development, particularly focusing on inhibition of the kinase activity. However, available activity assays are discontinuous, involving either radioactivity detection or coupling with antibodies. Here we describe a continuous and direct assay for LRRK2 kinase activity, combining a reported peptide sequence optimized for LRRK2 binding and an established strategy for fluorescence emission on magnesium ion chelation by phosphorylated peptides carrying an artificial amino acid. The assay was employed to evaluate apparent steady-state parameters for the wild type and two mutant forms of LRRK2 associated with Parkinson's disease as well as to probe the effects of GTP, GDP, and autophosphorylation on the kinase activity of the enzyme. Staurosporine was evaluated as an inhibitor of the wild-type enzyme. It is expected that this assay will aid in mechanistic investigations of LRRK2.
Related JoVE Video
LASSBio-1135: a dual TRPV1 antagonist and anti-TNF-alpha compound orally effective in models of inflammatory and neuropathic pain.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
LASSBio-1135 is an imidazo[1,2-a]pyridine derivative with high efficacy in screening models of nociception and inflammation, presumed as a weak COX-2 inhibitor. In order to tease out its mechanism of action, we investigated others possible target for LASSBio-1135, such as TNF-? and TRPV1, to better characterize it as a multitarget compound useful in the treatment of chronic pain. TRPV1 modulation was assessed in TRPV1-expressing Xenopus oocytes against capsaicin and low pH-induced current. Modulation of TNF-? production was evaluated in culture of macrophages stimulated with LPS. In vivo efficacy of LASSBio-1135 was investigated in carrageenan and partial sciatic ligation-induced thermal hyperalgesia and mechanical allodynia. Corroborating its previous demonstration of efficacy in a model of capsaicin-induced hyperalgesia, LASSBio-1135 blocks capsaicin-elicited currents in a non-competitive way with an IC50 of 580 nM as well as low pH-induced current at 50 µM. As an additional action, LASSBio-1135 inhibited TNF-? release in these cells stimulated by LPS with an IC50 of 546 nM by reducing p38 MAPK phosphorilation. Oral administration of 100 µmol x Kg(-1) LASSBio-1135 markedly reduced thermal hyperalgesia induced by carrageenan, however at 10 µmol x Kg(-1) only a partial reduction was observed at the 4th h. Neutrophil recruitment and TNF-? production after carrageenan stimulus was also inhibited by the treatment with LASSBio-1135. Modulating TRPV1 and TNF-? production, two key therapeutic targets of neuropathic pain, 100 µmol x Kg(-1) LASSBio-1135 was orally efficacious in reversing thermal hyperalgesia and mechanical allodynia produced by partial sciatic ligation 7-11 days after surgery without provoking hyperthermia, a common side effect of TRPV1 antagonists. In conclusion LASSBio-1135, besides being a weak COX-2 inhibitor, is a non-competitive TRPV1 antagonist and a TNF-? inhibitor. As a multitarget compound, LASSBio-1135 is orally efficacious in a model of neuropathic pain without presenting hyperthermia.
Related JoVE Video
Deciphering the cis-regulatory elements for XYR1 and CRE1 regulators in Trichoderma reesei.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
In this work, we report the in silico identification of the cis-regulatory elements for XYR1 and CRE1 proteins in the filamentous fungus Trichoderma reesei, two regulators that play a central role in the expression of cellulase genes. Using four datasets of condition-dependent genes from RNA-seq and RT-qPCR experiments, we performed unsupervised motif discovery and found two short motifs resembling the proposed binding consensus for XYR1 and CRE1. Using these motifs, we analysed the presence and arrangement of putative cis-regulatory elements recognized by both regulators and found that shortly spaced sites were more associated with XYR1- and CRE1-dependent promoters than single, high-score sites. Furthermore, the approach used here allowed the identification of the previously reported XYR1-binding sites from cel7a and xyn1 promoters, and we also mapped the potential target sequence for this regulator at the cel6a promoter that has been suggested but not identified previously. Additionally, seven other promoters (for cel7b, cel61a, cel61b, cel3c, cel3d, xyn3 and swo genes) presented a putative XYR1-binding site, and strong sites for CRE1 were found at the xyr1 and cel7b promoters. Using the cis-regulatory architectures nearly defined for XYR1 and CRE1, we performed genome-wide identification of potential targets for direct regulation by both proteins and important differences on their functional regulons were elucidated. Finally, we performed binding site mapping on the promoters of differentially expressed genes found in T. reesei mutant strains lacking xyr1 or cre1 and found that indirect regulation plays a key role on their signalling pathways. Taken together, the data provided here sheds new light on the mechanisms for signal integration mediated by XYR1 and CRE1 at cellulase promoters.
Related JoVE Video
Simultaneous analysis of p53 protein expression and cell proliferation in irradiated human lymphocytes by flow cytometry.
Dose Response
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
P53 protein has an intrinsic role in modulating the cellular response against DNA radioinduced damages and has been pointed out as an indirect indicator of individual radiosensitivity. The rate of cell proliferation is also a parameter that has been related to tissue sensitivity to radiation. However, this feature is yet understudied. In this context, the aim of this work was to employ Flow Cytometry (FC) for simultaneously assessing of p53 protein expression levels together with cellular proliferation rate of irradiated human lymphocytes. From in vitro irradiated human blood samples, mononuclear cells were isolated and labeled with Carboxylfluorescein Diacetate Succinimidyl Ester (CFSE) prior to phytohaemagglutinin (PHA) stimulation in culture for 96 hours. Cells were also labeled with anti-p53 monoclonal antibody PE-conjugated in order to analyze either proliferation rate or p53 expression levels by FC. It was verified a reduction in the proliferation rate of irradiated lymphocytes and, in parallel, a rise in the p53 expression levels, similar for quiescent and proliferating lymphocytes. The results emphasize the importance of the use of CFSE-stained lymphocytes in assays associated to proliferation rate and the use of this methodology in several studies, such as for evaluating individual radiosensitivity.
Related JoVE Video
The Effects of Two Different Doses of Calcium Lactate on Blood pH, Bicarbonate and Repeated High-Intensity Exercise Performance.
Int J Sport Nutr Exerc Metab
PUBLISHED: 11-25-2013
Show Abstract
Hide Abstract
We investigated the effects of low- and high-dose calcium lactate supplementation on blood pH and bicarbonate (Study A) and on repeated high-intensity performance (Study B). In Study A, ten young, physically active men (age: 24 ± 2.5 years; weight: 79.2 ± 9.45 kg; height: 1.79 ± 0.06 m) were assigned to acutely receive three different treatments, in a crossover fashion: high-dose calcium lactate (HD: 300 mg.kg-1 body mass), low-dose calcium lactate (LD: 150 mg.kg-1 body mass) and placebo (PL). During each visit, participants received one of these treatments and were assessed for blood pH and bicarbonate 0, 60, 90, 120, 150, 180 and 240 minutes following ingestion. In Study B, twelve young male participants (age: 26 ± 4.5 years; weight: 82.0 ± 11.0 kg; height: 1.81 ± 0.07 m) received the same treatments of Study A. Ninety minutes after ingestion, participants underwent 3 bouts of the upper-body Wingate test and were assessed for blood pH and bicarbonate 0 and 90 min following ingestion and immediately after exercise. In Study A, both HD and LD promoted slight but significant increases in blood bicarbonate (31.47 ± 1.57 and 31.69 ± 1.04 mmol.l-1, respectively) and pH levels (7.36 ± 0.02 and 7.36 ± 0.01, respectively), with no effect of PL. In Study B, total work done, peak power, mean power output were not affected by treatments. In conclusion, low- and high-dose calcium lactate supplementation induced similar, yet very discrete, increases in blood pH and bicarbonate, which were not sufficiently large to improve repeated high-intensity performance.
Related JoVE Video
Conducting MoS2 Nanosheets as Catalysts for Hydrogen Evolution Reaction.
Nano Lett.
PUBLISHED: 11-21-2013
Show Abstract
Hide Abstract
We report chemically exfoliated MoS2 nanosheets with a very high concentration of metallic 1T phase using a solvent free intercalation method. After removing the excess of negative charges from the surface of the nanosheets, highly conducting 1T phase MoS2 nanosheets exhibit excellent catalytic activity toward the evolution of hydrogen with a notably low Tafel slope of 40 mV/dec. By partially oxidizing MoS2, we found that the activity of 2H MoS2 is significantly reduced after oxidation, consistent with edge oxidation. On the other hand, 1T MoS2 remains unaffected after oxidation, suggesting that edges of the nanosheets are not the main active sites. The importance of electrical conductivity of the two phases on the hydrogen evolution reaction activity has been further confirmed by using carbon nanotubes to increase the conductivity of 2H MoS2.
Related JoVE Video
Intranasal Administration of Perillyl Alcohol Activates Peripheral and Bronchus-Associated Immune System In Vivo.
Arch. Immunol. Ther. Exp. (Warsz.)
PUBLISHED: 11-05-2013
Show Abstract
Hide Abstract
Perillyl alcohol (POH) presents antitumoral activity but clinical application is hampered by adverse effects following oral administration. This work aimed to verify the cytotoxic effect of intranasal POH administration in the histology of lung, liver, brain; the cellularity and function of peripheral and bronchoalveolar-associated immune system. C57 adult mice received 1-min inhalation with POH, vehicle 70 % ethanol or saline buffer, once (84 ?g/day) or twice (164 ?g/day) during five consecutive days, and were killed 72 h after treatment. Spleen, cervical and mesenteric lymph nodes were removed for (3)H-thymidine proliferation assay, leukocyte cellularity and flow cytometry analysis. Peripheral blood and bronchoalveolar lavage cells were collected to assess cellularity and immunoglobulin (IgA, IgM) levels. Intranasal POH did not alter body weight or liver, brain and lung morphology, but increased splenocyte and cervical lymph node cell proliferation, and IgM production without altering peripheral lymphocyte subsets. Treatment also increased the percentage of alveolar macrophages (83 %) and IgA-producing lymphocytes (15 %), a pattern characteristic of activated bronchoalveolar innate immune system. Intranasal administration of POH activated peripheral immune system and innate immunity of bronchus-associated lymphoid tissue, thus suggesting a possible role for POH as a chemotherapeutic drug also in pathological processes affecting the lung.
Related JoVE Video
Increased energy/activity, not mood changes, is the core feature of mania.
J Affect Disord
PUBLISHED: 09-17-2013
Show Abstract
Hide Abstract
In the Diagnostic and Statistical Manual of Mental Disorders, 5th edition, the occurrence of increased energy/activity and elation of mood or irritability became necessary symptoms for the diagnosis of an episode of mania or hypomania.
Related JoVE Video
Genome Sequence of an Epidemic Isolate of Mycobacterium abscessus subsp. bolletii from Rio de Janeiro, Brazil.
Genome Announc
PUBLISHED: 08-17-2013
Show Abstract
Hide Abstract
Multiple isolates of Mycobacterium abscessus subsp. bolletii, collectively called BRA100, were associated with outbreaks of postsurgical skin infections across various regions of Brazil from 2003 to 2009. We announce the draft genome sequence of a newly sequenced BRA100 strain, M. abscessus subsp. bolletii CRM-0020, isolated from a patient in Rio de Janeiro, Brazil.
Related JoVE Video
Engineering Multicellular Logic in Bacteria with Metabolic Wires.
ACS Synth Biol
PUBLISHED: 07-22-2013
Show Abstract
Hide Abstract
Aromatic biodegradation pathways of environmental bacteria are vast sources of matching trios of enzymes, substrates and regulators that can be refactored to run logic operations through cell-to-cell communication. As a proof of concept, the connection between two Pseudomonas putida strains using benzoic acid as the wiring molecule is presented. In this system, a sender strain harboring the TOL pathway for biodegradation of aromatics processed toluene as input and generated benzoate as the output signal. Diffusion of such metabolic intermediate to the medium was then sensed by a second strain (the receiver) that used benzoate as input for a new logic gate producing a visual output (i.e., light emission). The setup was functional irrespective of whether sender and receiver cells were in direct contact or in liquid culture. These results highlight the potential of environmental metabolic pathways as sources of building blocks for the engineering of multicellular logic in prokaryotic systems.
Related JoVE Video
Efficient oxygen evolution reaction catalyzed by low-density Ni-doped Co3O4 nanomaterials derived from metal-embedded graphitic C3N4.
Chem. Commun. (Camb.)
PUBLISHED: 07-19-2013
Show Abstract
Hide Abstract
A synthetic route to low-density porous Ni-doped Co3O4 nanomaterials that show stable and superior electrocatalytic activity for O2 evolution reaction is reported. The materials are prepared via thermal treatment of "pre-synthesized" metal ions-embedded graphitic C3N4-based polymers.
Related JoVE Video
Phylogenomics of Brazilian epidemic isolates of Mycobacterium abscessus subsp. bolletii reveals relationships of global outbreak strains.
Infect. Genet. Evol.
PUBLISHED: 07-15-2013
Show Abstract
Hide Abstract
Rapidly growing, non-tuberculous mycobacteria (NTM) in the Mycobacterium abscessus (MAB) species are emerging pathogens that cause various diseases including skin and respiratory infections. The species has undergone recent taxonomic nomenclature refinement, and is currently recognized as two subspecies, M. abscessus subsp. abscessus (MAB-A) and M. abscessus subsp. bolletii (MAB-B). The recently reported outbreaks of MAB-B in surgical patients in Brazil from 2004 to 2009 and in cystic fibrosis patients in the United Kingdom (UK) in 2006 to 2012 underscore the need to investigate the genetic diversity of clinical MAB strains. To this end, we sequenced the genomes of two Brazilian MAB-B epidemic isolates (CRM-0019 and CRM-0020) derived from an outbreak of skin infections in Rio de Janeiro, two unrelated MAB strains from patients with pulmonary infections in the United States (US) (NJH8 and NJH11) and one type MAB-B strain (CCUG 48898) and compared them to 25 publically available genomes of globally diverse MAB strains. Genome-wide analyses of 27,598 core genome single nucleotide polymorphisms (SNPs) revealed that the two Brazilian derived CRM strains are nearly indistinguishable from one another and are more closely related to UK outbreak isolates infecting CF patients than to strains from the US, Malaysia or France. Comparative genomic analyses of six closely related outbreak strains revealed geographic-specific large-scale insertion/deletion variation that corresponds to bacteriophage insertions and recombination hotspots. Our study integrates new genome sequence data with existing genomic information to explore the global diversity of infectious M. abscessus isolates and to compare clinically relevant outbreak strains from different continents.
Related JoVE Video
Immediate effect of a single anteroposterior talus mobilization on dorsiflexion range of motion in participants with orthopedic dysfunction of the ankle and foot.
J Manipulative Physiol Ther
PUBLISHED: 07-11-2013
Show Abstract
Hide Abstract
The purpose of this study was to determine the immediate effects of a single anteroposterior mobilization of the talus on the active dorsiflexion range of motion (ROM) in participants with different orthopedic foot and ankle injuries.
Related JoVE Video
Proximal opening-wedge osteotomy of the first metatarsal for moderate and severe hallux valgus using low profile plates.
Foot Ankle Surg
PUBLISHED: 06-30-2013
Show Abstract
Hide Abstract
To present the results of the treatment of hallux valgus with the proximal metatarsal opening wedge (PMOW) osteotomy using two different screw and plate systems.
Related JoVE Video
The genome of Anopheles darlingi, the main neotropical malaria vector.
Osvaldo Marinotti, Gustavo C Cerqueira, Luiz Gonzaga Paula de Almeida, Maria Inês Tiraboschi Ferro, Élgion Lúcio da Silva Loreto, Arnaldo Zaha, Santuza M R Teixeira, Adam R Wespiser, Alexandre Almeida E Silva, Aline Daiane Schlindwein, Ana Carolina Landim Pacheco, Artur Luiz da Costa da Silva, Brenton R Graveley, Brian P Walenz, Bruna de Araujo Lima, Carlos Alexandre Gomes Ribeiro, Carlos Gustavo Nunes-Silva, Carlos Roberto de Carvalho, Célia Maria de Almeida Soares, Claudia Beatriz Afonso de Menezes, Cleverson Matiolli, Daniel Caffrey, Demetrius Antonio M Araújo, Diana Magalhaes de Oliveira, Douglas Golenbock, Edmundo Carlos Grisard, Fabiana Fantinatti-Garboggini, Fabíola Marques de Carvalho, Fernando Gomes Barcellos, Francisco Prosdocimi, Gemma May, Gilson Martins de Azevedo Junior, Giselle Moura Guimarães, Gustavo Henrique Goldman, Itácio Q M Padilha, Jacqueline da Silva Batista, Jesus Aparecido Ferro, José M C Ribeiro, Juliana Lopes Rangel Fietto, Karina Maia Dabbas, Louise Cerdeira, Lucymara Fassarella Agnez-Lima, Marcelo Brocchi, Marcos Oliveira de Carvalho, Marcus de Melo Teixeira, Maria de Mascena Diniz Maia, Maria Helena S Goldman, Maria Paula Cruz Schneider, Maria Sueli Soares Felipe, Mariangela Hungria, Marisa Fabiana Nicolás, Maristela Pereira, Martín Alejandro Montes, Mauricio E Cantão, Michel Vincentz, Míriam Silva Rafael, Neal Silverman, Patrícia Hermes Stoco, Rangel Celso Souza, Renato Vicentini, Ricardo Tostes Gazzinelli, Rogério de Oliveira Neves, Rosane Silva, Spartaco Astolfi-Filho, Talles Eduardo Ferreira Maciel, Turán P Urményi, Wanderli Pedro Tadei, Erney Plessmann Camargo, Ana Tereza Ribeiro de Vasconcelos.
Nucleic Acids Res.
PUBLISHED: 06-12-2013
Show Abstract
Hide Abstract
Anopheles darlingi is the principal neotropical malaria vector, responsible for more than a million cases of malaria per year on the American continent. Anopheles darlingi diverged from the African and Asian malaria vectors ?100 million years ago (mya) and successfully adapted to the New World environment. Here we present an annotated reference A. darlingi genome, sequenced from a wild population of males and females collected in the Brazilian Amazon. A total of 10 481 predicted protein-coding genes were annotated, 72% of which have their closest counterpart in Anopheles gambiae and 21% have highest similarity with other mosquito species. In spite of a long period of divergent evolution, conserved gene synteny was observed between A. darlingi and A. gambiae. More than 10 million single nucleotide polymorphisms and short indels with potential use as genetic markers were identified. Transposable elements correspond to 2.3% of the A. darlingi genome. Genes associated with hematophagy, immunity and insecticide resistance, directly involved in vector-human and vector-parasite interactions, were identified and discussed. This study represents the first effort to sequence the genome of a neotropical malaria vector, and opens a new window through which we can contemplate the evolutionary history of anopheline mosquitoes. It also provides valuable information that may lead to novel strategies to reduce malaria transmission on the South American continent. The A. darlingi genome is accessible at www.labinfo.lncc.br/index.php/anopheles-darlingi.
Related JoVE Video
A virtual imaging platform for multi-modality medical image simulation.
IEEE Trans Med Imaging
PUBLISHED: 06-12-2013
Show Abstract
Hide Abstract
This paper presents the Virtual Imaging Platform (VIP), a platform accessible at http://vip.creatis.insa-lyon.fr to facilitate the sharing of object models and medical image simulators, and to provide access to distributed computing and storage resources. A complete overview is presented, describing the ontologies designed to share models in a common repository, the workflow template used to integrate simulators, and the tools and strategies used to exploit computing and storage resources. Simulation results obtained in four image modalities and with different models show that VIP is versatile and robust enough to support large simulations. The platform currently has 200 registered users who consumed 33 years of CPU time in 2011.
Related JoVE Video
Surgical research in the north and northeast of Brazil.
Acta Cir Bras
PUBLISHED: 06-08-2013
Show Abstract
Hide Abstract
To evaluate the scientific production from the surgical areas from the State University of Para (UEPA), Federal Universities of Para (UFPA) and of Paraiba (UFPB), Brazil.
Related JoVE Video
Isolation and Characterization of Phytotoxic Compounds Produced by Streptomyces sp. AMC 23 from Red Mangrove (Rhizophora mangle).
Appl. Biochem. Biotechnol.
PUBLISHED: 06-05-2013
Show Abstract
Hide Abstract
Natural products produced by microorganisms have been utilized as sources of new drugs possessing a wide range of agrochemical and pharmacological activities. During our research on Actinomycetes from Brazilian mangroves, the ethyl acetate extract of Streptomyces sp. AMC 23 isolated from the red mangrove (Rhizophora mangle) rhizosphere produced a highly active compound against the microalga Chlorella vulgaris, often used to assess the phytotoxic activity. As a result, the bioassay-guided fractionation led to the isolation of the mixture of the known compounds bafilomycin B1 and bafilomycin B2. The chemical structures of bafilomycin B1 and bafilomycin B2 were established based on their spectroscopic data by infrared (IR), mass spectrometry (MS), (1)H nuclear magnetic resonance (NMR), gradient-enhanced heteronuclear multiple quantum coherence (gHMQC), and gradient-enhanced heteronuclear multiple-bond connectivity (gHMBC) as well as comparison with reference data from the literature. Moreover, it was also possible to identify other bafilomycins using non-chromatographic-dependent techniques (Tandem mass spectrometry). Additionally, this is the first report on the phytotoxic activity of bafilomycin B1.
Related JoVE Video
Enhanced catalytic activity in strained chemically exfoliated WS? nanosheets for hydrogen evolution.
Nat Mater
PUBLISHED: 05-29-2013
Show Abstract
Hide Abstract
Efficient evolution of hydrogen through electrocatalysis at low overpotentials holds tremendous promise for clean energy. Hydrogen evolution can be easily achieved by electrolysis at large potentials that can be lowered with expensive platinum-based catalysts. Replacement of Pt with inexpensive, earth-abundant electrocatalysts would be significantly beneficial for clean and efficient hydrogen evolution. To this end, promising results have been reported using 2H (trigonal prismatic) XS? (where X? = ?Mo or W) nanoparticles with a high concentration of metallic edges. The key challenges for XS? are increasing the number and catalytic activity of active sites. Here we report monolayered nanosheets of chemically exfoliated WS? as efficient catalysts for hydrogen evolution with very low overpotentials. Analyses indicate that the enhanced electrocatalytic activity of WS? is associated with the high concentration of the strained metallic 1T (octahedral) phase in the as-exfoliated nanosheets. Our results suggest that chemically exfoliated WS? nanosheets are interesting catalysts for hydrogen evolution.
Related JoVE Video
Efficient metal-free electrocatalysts for oxygen reduction: polyaniline-derived N- and O-doped mesoporous carbons.
J. Am. Chem. Soc.
PUBLISHED: 05-16-2013
Show Abstract
Hide Abstract
The oxygen reduction reaction (ORR)-one of the two half-reactions in fuel cells-is one of the bottlenecks that has prevented fuel cells from finding a wide range of applications today. This is because ORR is inherently a sluggish reaction; it is also because inexpensive and sustainable ORR electrocatalysts that are not only efficient but also are based on earth-abundant elements are hard to come by. Herein we report the synthesis of novel carbon-based materials that can contribute to solving these challenges associated with ORR. Mesoporous oxygen- and nitrogen-doped carbons were synthesized from in situ polymerized mesoporous silica-supported polyaniline (PANI) by carbonization of the latter, followed by etching away the mesoporous silica template from it. The synthetic method also allowed the immobilization of different metals such as Fe and Co easily into the system. While all the resulting materials showed outstanding electrocatalytic activity toward ORR, the metal-free, PANI-derived mesoporous carbon (dubbed PDMC), in particular, exhibited the highest activity, challenging conventional paradigms. This unprecedented activity by the metal-free PDMC toward ORR was attributed to the synergetic activities of nitrogen and oxygen (or hydroxyl) species that were implanted in it by PANI/mesoporous silica during pyrolysis.
Related JoVE Video
Rapid detection and differentiation of mycobacterial species using a multiplex PCR system.
Rev. Soc. Bras. Med. Trop.
PUBLISHED: 05-06-2013
Show Abstract
Hide Abstract
The early diagnosis of mycobacterial infections is a critical step for initiating treatment and curing the patient. Molecular analytical methods have led to considerable improvements in the speed and accuracy of mycobacteria detection.
Related JoVE Video
Transcriptomic fingerprinting of Pseudomonas putida under alternative physiological regimes.
Environ Microbiol Rep
PUBLISHED: 05-01-2013
Show Abstract
Hide Abstract
Pseudomonas putida?KT2440 is a metabolically versatile soil bacterium useful both as a model biodegradative organism and as a host of catalytic activities of biotechnological interest. In this report, we present the high-resolution transcriptome of P.?putida cultured on different carbon sources as revealed by deep sequencing of the corresponding RNA pools. Examination of the data from growth on substrates that are processed through distinct pathways (glucose, fructose, succinate and glycerol) revealed that ??20% of the P.?putida genome is differentially expressed depending on the ensuing physiological regime. Changes affected not only metabolic genes but also a suite of global regulators, e.g. the rpoS sigma subunit of RNA polymerase, various cold-shock proteins and the three HU histone-like proteins. Specifically, the genes encoding HU subunit variants hupA, hupB and hupN drastically altered their expression levels (and thus their ability to form heterodimeric combinations) under the diverse growth conditions. Furthermore, we found that two small RNAs, crcZ and crcY, known to inhibit the Crc protein that mediates catabolite repression in P.?putida, were both down-regulated by glucose. The raw transcriptomic data generated in this work is made available to the community through the Gene Expression Omnibus database.
Related JoVE Video
[Prevalence of blood donation and associated factors in Florianópolis, Southern Brazil: a population-based study].
Cad Saude Publica
PUBLISHED: 04-25-2013
Show Abstract
Hide Abstract
The main objectives of this study were to estimate the prevalence of blood donation and to identify associated factors. A populated-based cross-sectional study was carried out (n = 1,720) including subjects 20 to 59 years of age in Florianópolis, Santa Catarina State, Brazil. Blood donation any time in life and in the previous year was reported by 30.6% and 6.2% of respondents, respectively. Among the latter, 31.8% reported repeat donation (at least twice in the previous year), 80.4% reported voluntary donation, and 15.9% replacement donation. Multivariate Poisson regression (p < 0.05) showed that male gender, black or brown self-reported skin color, higher age group, higher educational level, and living with other blood donors were all associated with ever donating blood, whereas younger age, single marital status, and higher educational level were associated with donating in the previous year. We found higher prevalence of blood donation in the previous year and higher percentage of young donors in this group, as well as lower percentage of replacement donors than previously reported in Brazil.
Related JoVE Video
Effect of hospital disinfectants on spores of clinical Brazilian Clostridium difficile strains.
Anaerobe
PUBLISHED: 04-12-2013
Show Abstract
Hide Abstract
The aim of this study was to evaluate the sporicidal activity of hospital disinfectants against spores of two Brazilian Clostridium difficile ribotypes and the BI/NAP1/027. Our results showed that CloroRio(®) and Cidex Opa(®) were the most efficient agents for eliminating spores of C difficile.
Related JoVE Video
Assessing rheumatoid arthritis disease activity with ultrasound.
Clin. Rheumatol.
PUBLISHED: 04-10-2013
Show Abstract
Hide Abstract
In practice, composite indices are used for rheumatoid arthritis (RA) disease activity evaluation. Despite valid and widely used, not rarely composite indices miss accuracy. Ultrasound (US) is more precise than clinical examination in synovitis appraisal. US-based disease activity estimation depends on the detection of synovitis. The most common synovitis abnormalities are proliferation, effusion, and neoangiogenesis. Gray scale ultrasound identifies synovial hypertrophy and effusion with its good soft tissue contrast. Additionally, power Doppler ultrasound depicts neoangiogenesis within synovia, remarking local inflammation. Several studies have combined local US findings to develop a patient level disease activity index. Most of them summed selected joint scores in an overall score of disease activity and evaluated its correlation with clinical composite indexes. To be incorporated into clinical practice, an overall US score must have some fundamental characteristics such as reproducibility, viability, and sensitivity to change over time. In global US score development, finding the joints that truly estimate individual disease activity is highly challenging. This article presents an up-to-date literature review on assessing RA disease activity with US and depicts the challenges in finding the perfect global US score.
Related JoVE Video
Plasmodium vivax malaria: related factors to severity in the State of Maranhão, Brazil.
Rev. Soc. Bras. Med. Trop.
PUBLISHED: 04-09-2013
Show Abstract
Hide Abstract
Malaria caused by Plasmodium vivax species has shown signs of severity, recorded with increasing frequency in the medical literature. This study aimed to characterize the signs of severe malaria by Plasmodium vivax in the State of Maranhão, Brazil.
Related JoVE Video
Super giant squamous cell carcinomas.
Cutis
PUBLISHED: 03-22-2013
Show Abstract
Hide Abstract
Squamous cell carcinomas (SCCs) larger than 2 cm in diameter are associated with a greater risk for disfigurement, local recurrence, and metastasis. Giant SCCs are rare. They may develop near aesthetically and functionally important structures and infiltrate them. Treatment may be difficult with the need for large tissue resections and complex surgical reconstruction, with important repercussions on the quality of life of patients. We report 2 cases of super giant SCCs with long-standing evolution that were successfully treated with surgical excision and followed for 24 months posttreatment without signs of local recurrence and/or metastasis.
Related JoVE Video
Selection of reliable reference genes for quantitative real-time polymerase chain reaction studies in maize grains.
Plant Cell Rep.
PUBLISHED: 03-08-2013
Show Abstract
Hide Abstract
The stability of candidate reference genes was evaluated in maize landrace varieties and during multiple grain developmental stages to evaluate the expression of carotenoid-related genes by RT-qPCR for application to maize biofortification. Vitamin A deficiency affects millions of children worldwide; therefore, increasing the content of vitamin A precursors in maize grains is of interest. The study of the expression of genes involved in the carotenoid biosynthetic pathway in maize grains has provided useful information for metabolic engineering approaches. However, reliable results using real-time quantitative polymerase chain reaction (RT-qPCR) experiments are dependent on the use of the appropriate reference genes. In this study, we utilized geNorm and NormFinder softwares to identify the most stably expressed candidate reference genes in samples from seven stages of grain development and from eight landrace varieties. The results of the analysis performed using geNorm indicated that tubulin (TUB) and actin (ACT) were the most suitable reference genes among all experimental conditions, while glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) showed the least stability. The same result was obtained with the NormFinder software. The minimum number of genes required in each experimental condition to normalize the gene expression data was also determined by geNorm. The expression of phytoene synthase gene (PSY1), the first enzyme in the carotenoid biosynthetic pathway, was overestimated when the least stable candidate gene (GAPDH) was used as the internal control instead of the most stable gene pair (ACT + TUB), thus highlighting the importance of validating reference genes before conducting a RT-qPCR experiment to obtain accurate results. This study is the first survey of the stability of genes for use as reference genes to normalize RT-qPCR data from maize landraces during multiple stages of grain development.
Related JoVE Video
Evaluation of Th17 related cytokines associated with clinical and laboratorial parameters in sickle cell anemia patients with leg ulcers.
Cytokine
PUBLISHED: 03-04-2013
Show Abstract
Hide Abstract
Leg ulcers (LUs) represent one of the main causes of morbidity in sickle cell anemia (SCA). This manifestation has been related to hemolysis, infections predisposition and inflammation that leads cytokines secretion. In this context, our study aimed to evaluate Th17 related cytokines (IL-6, IL-17A, IL-22 and IL-23) in serum and peripheral mononuclear cells culture supernatants with and without lymphoproliferative stimulation (anti-human CD3 and anti-human CD28). The cytokines levels were also correlated to clinical, hematological and biochemical parameters in SCA patients with and without LUs history (SCALU and SCAWH) as well as in healthy controls. In SCALU patients, high levels of IL-17A were associated with absence of acute chest syndrome (ACS, p=0.0328). The other clinical parameters analyzed (osteonecrosis, stroke, priapism, splenectomy and blood transfusions history) were not significantly related with other cytokine levels. In SCALU patients was also observed that IL-17A increased levels were associated with high levels of LDH (p=0.0130), the same association pattern was found for IL-6 (0.0160) and IL-22 (p=0.0165) in the SCALU group. Interestingly, we did not find statistical correlations with these parameters in SCAWH group. The other hematological parameters (hemoglobin, leucocyte and reticulocyte count) and indirect bilirrubin did not show any correlation with analyzed cytokines in both groups. So, for the first time, we show that IL-17A present in SCALU patients may exert a preventive role in the ACS development. Furthermore, IL-6, IL-17A and IL-22 accompanied the LDH levels only in SCALU patients suggesting to serve as additional markers of hemolysis or to be related with immunity response against extracellular pathogens.
Related JoVE Video
The Biginelli reaction with an imidazolium-tagged recyclable iron catalyst: kinetics, mechanism, and antitumoral activity.
Chemistry
PUBLISHED: 03-04-2013
Show Abstract
Hide Abstract
The present work describes the synthesis, characterization, and application of a new ion-tagged iron catalyst. The catalyst was employed in the Biginelli reaction with impressive performance. High yields have been achieved when the reaction was carried out in imidazolium-based ionic liquids (BMI?PF6, BMI?NTf2, and BMI?BF4), thus showing that the ionic-liquid effects play a role in the reaction. Moreover, the ion-tagged catalyst could be recovered and reused up to eight times without any noticeable loss in activity. Mechanistic studies performed by using high-resolution electrospray-ionization quadrupole-time-of-flight mass (HR-EI-QTOF) spectrometry and kinetic experiments indicate only one reaction pathway and rule out the other two possibilities under the development conditions. The theoretical calculations are in accordance with the proposed mechanism of action of the iron catalyst. Finally, the 37 dihydropyrimidinone derivatives, products of the Biginelli reaction, had their cytotoxicity evaluated in assays against MCF-7 cancer cell linages with encouraging results of some derivatives, which were virtually non-toxic against healthy cell linages (fibroblasts).
Related JoVE Video
Is migraine a complicating factor for evidence-based therapy for masticatory myofascial pain? A case-control study.
Oral Surg Oral Med Oral Pathol Oral Radiol
PUBLISHED: 02-20-2013
Show Abstract
Hide Abstract
This study aimed to assess the response to conservative treatment of pain in patients diagnosed with masticatory myofascial pain (MMP) with or without migraine.
Related JoVE Video
[Hughes-Stovin syndrome: Report of one case].
Rev Med Chil
PUBLISHED: 02-08-2013
Show Abstract
Hide Abstract
We report a 25yearold mole admitted to a critical care unit for fever, lung opacities and acute respiratory failure. A chest angio-CAT sean showed multiple pulmonary artery aneurysms. A deep venous thrombosis of both lower limbs was also documented. A Hughes-Stovin syndrome was postulated. An inferior vena cava filter was placed. The patient received antimicrobial therapy subsiding fever and respiratory failure. Subsequently, he was treated with intravenous and oral steroids and one dose of cyclophosphamide. The patient was discharged in good conditions fifteen days after admission.
Related JoVE Video
Exercise attenuates renal dysfunction with preservation of myocardial function in chronic kidney disease.
PLoS ONE
PUBLISHED: 02-07-2013
Show Abstract
Hide Abstract
Previous studies have suggested that exercise improves renal and cardiac functions in patients with chronic kidney disease. The aim of this study was to evaluate the effects of long-term aerobic swimming exercise with overload on renal and cardiac function in rats with 5/6 nefrectomy (5/6Nx). Eight Wistar rats were placed into 4 groups: Control (C), Control+Exercise (E), Sedentary 5/6Nx (NxS) and 5/6Nx+Exercise (NxE). The rats were subjected to swimming exercise sessions with overload for 30 min five days per week for five weeks. Exercise reduced the effect of 5/6Nx on creatinine clearance compared to the NxS group. In addition, exercise minimized the increase in mean proteinuria compared to the NxS group (96.9±10.0 vs. 51.4±9.9 mg/24 h; p<0.05). Blood pressure was higher in the NxS and NxE groups compared to the C and E groups (216±4 and 178±3 vs. 123±2 and 124±2 mm Hg, p<0.05). In the 200 glomeruli that were evaluated, the NxS group had a higher sclerosis index than did the NxE group (16% vs. 2%, p<0.05). Echocardiography demonstrated a higher anterior wall of the left ventricle (LV) in diastole in the NxS group compared with the C, E and NxE groups. The NxS group also had a higher LV posterior wall in diastole and systole compared with the E group. The developed isometric tension in Lmax of the heart papillary muscle was lower in the NxS group compared with the C, E and NxE groups. These results suggested that exercise in 5/6Nx animals might reduce the progression of renal disease and lessen the cardiovascular impact of a reduction in renal mass.
Related JoVE Video
Characterization of non-tuberculosis mycobacteria by neutron radiography.
Appl Radiat Isot
PUBLISHED: 02-06-2013
Show Abstract
Hide Abstract
The genus Mycobacterium shares many characteristics with Corynebacterium and Actinomyces genera, among which the genomic guanine plus cytosine content and the production of long branched-chain fatty acids, known as mycolic acids are enhanced. Growth rate and optimal temperature of mycobacteria are variable. The genus comprises more than 140 known species; however Mycobacterium fortuitum, a fast growing nontuberculous mycobacterium, is clinically significant, because it has been associated to several lesions following surgery procedures such as liposuction, silicone breast and pacemaker implants, exposure to prosthetic materials besides sporadic lesions in the skin, soft tissues and rarely lungs. The objective of the present study is to reduce the time necessary for M. fortuitum characterization based on its morphology and the use of the neutron radiography technique substituting the classical biochemical assays. We also aim to confirm the utility of dendrimers as boron carriers. The samples were sterilized through conventional protocols using 10% formaldehyde. In the incubation process, two solutions with different molar ratios (10:1 and 20:1) of sodium borate and PAMAM G4 dendrimer and also pure sodium borate were used. After doping and sterilization procedures, the samples were deposited on CR-39 sheets, irradiated with a 4.6×10(5) n/cm(2)s thermal neutron flux for 30 min, from the J-9 irradiation channel of the Argonauta IEN/CNEN reactor. The images registered in the CR-39 were visualized in a Nikon E400 optical transmission microscope and captured by a Nikon Coolpix 995 digital camera. Developing the nuclear tracks registered in the CR-39 allowed a 1000× enlargement of mycobacterium images, facilitating their characterization, the use of more sophisticated equipment not being necessary. The use of neutron radiography technique reduced the time necessary for characterization. Doping with PAMAM dendrimer improved the visualization of NTM in neutron radiography images.
Related JoVE Video
Cratylia mollis 1, 4 lectin: a new biotechnological tool in IL-6, IL-17A, IL-22, and IL-23 induction and generation of immunological memory.
Biomed Res Int
PUBLISHED: 01-30-2013
Show Abstract
Hide Abstract
Cratylia mollis lectin has already established cytokine induction in Th1 and Th2 pathways. Thereby, this study aimed to evaluate Cramoll 1, 4 in IL-6, IL-17A, IL-22, and IL-23 induction as well as analyze immunologic memory mechanism by reinducing lymphocyte stimulation. Initially we performed a screening in cultured splenocytes where Cramoll 1, 4 stimulated IL-6 production 5x more than ConA (P < 0.05). The same behavior was observed with IL-22 where the increase was greater than 4x. Nevertheless, IL-17A induction was similar for both lectins. In PBMCs, the same splenocytes course was observed for IL-6 and IL-17A. Concerning the stimulation of IL-22 and IL-23 Cramoll 1, 4 was more efficient than ConA in cytokines stimulation mainly in IL-23 (P < 0.01). Analyzing reinduced lymphocyte stimulation, IL-17A production was higher (P < 0.001) when the first stimulus was realized with Cramoll 1, 4 at 1 ? g/mL and the second at 5 ? g/mL. IL-22 shows significant differences (P < 0.01) at the same condition. Nevertheless, IL-23 revels the best response when the first stimuli was realized with Cramoll1, 4 at 100 ng/mL and the second with 5 ? g/mL. We conclude that the Cramoll 1, 4 is able to induce IL-6, IL-17A, IL-22, and IL-23 cytokines in vitro better than Concavalin A, besides immunologic memory generation, being a potential biotechnological tool in Th17 pathway studies.
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.