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.
Using WatchMinder to Increase the On-Task Behavior of Students with Autism Spectrum Disorder.
J Autism Dev Disord
PUBLISHED: 11-08-2014
Show Abstract
Hide Abstract
This study assessed the use of WatchMinder™, a vibrating prompt watch, and self-graphing on the on-task behavior of students with autism spectrum disorder in an elementary special education setting. Using a multiple baseline across subjects design, results showed an immediate increase in on-task behavior when the intervention was introduced. Participants maintained high levels of on-task behavior during the follow-up phase. Implications for expanded self-monitoring treatment packages are discussed.
Related JoVE Video
Antibiotic prescribing for respiratory infections: a cross-sectional analysis of the ReCEnT study exploring the habits of early-career doctors in primary care.
Fam Pract
PUBLISHED: 11-02-2014
Show Abstract
Hide Abstract
Antibiotic resistance is among the most important current public health issues. Most antibiotics are prescribed in primary care. There is strong consensus that they are overprescribed, especially for conditions such as upper respiratory tract infections (URTI) and acute bronchitis, where they provide limited benefit. Interventions to alter prescribing patterns have shown limited effect. Trainees in family practice may be an appropriate target, as their prescribing habits are still developing.
Related JoVE Video
How we use patient encounter data for reflective learning in family medicine training.
Med Teach
PUBLISHED: 10-15-2014
Show Abstract
Hide Abstract
Abstract Introduction: Consulting with patients is the core learning activity of Australian family medicine (general practice/GP) training, providing a rich source of reflective learning for trainees. We have developed a reflective learning program for postgraduate vocational trainees based on clinical encounters. Methods: The Registrar Clinical Encounters in Training (ReCEnT) program is an educational program documenting GP trainees' consultations in five Australian GP training providers. Trainees record patient demographics, consultation details, problems managed, management practices and educational factors from sixty consecutive consultations per six-month training term. Trainees receive a detailed feedback report comparing individual data to aggregated trainee data and national GP data. Results: The patient encounter system provides multiple opportunities for reflective learning across a number of domains of exposure and practice. Reflection can occur during completion of the encounter form; as self-reflection on the feedback report; as facilitated reflection with the GP trainer and medical educator; and as part of integration of data into teaching. We have identified areas for further development, including enhancing the reflective skills of trainees and trainers. Conclusion: The ReCEnT patient encounter program provides a rich platform for reflective learning for vocational trainees and supports development of skills in lifelong learning.
Related JoVE Video
Technology, normalisation and male sex work.
Cult Health Sex
PUBLISHED: 09-13-2014
Show Abstract
Hide Abstract
Technological change, particularly the growth of the Internet and smart phones, has increased the visibility of male escorts, expanded their client base and diversified the range of venues in which male sex work can take place. Specifically, the Internet has relocated some forms of male sex work away from the street and thereby increased market reach, visibility and access and the scope of sex work advertising. Using the online profiles of 257 male sex workers drawn from six of the largest websites advertising male sexual services in Australia, the role of the Internet in facilitating the normalisation of male sex work is discussed. Specifically we examine how engagement with the sex industry has been reconstituted in term of better informed consumer-seller decisions for both clients and sex workers. Rather than being seen as a 'deviant' activity, understood in terms of pathology or criminal activity, male sex work is increasingly presented as an everyday commodity in the market place. In this context, the management of risks associated with sex work has shifted from formalised social control to more informal practices conducted among online communities of clients and sex workers. We discuss the implications for health, legal and welfare responses within an empowerment paradigm.
Related JoVE Video
Problems managed by Australian general practice trainees: results from the ReCenT (Registrar Clinical Encounters in Training) study.
Educ Prim Care
PUBLISHED: 09-09-2014
Show Abstract
Hide Abstract
BACKGROUND Previous studies have found that general practitioner (GP) trainees (registrars) see a different spectrum of clinical problems compared to trainers, including less chronic disease and more acute minor illness. Our aim was to describe the case mix of first-term Australian GP trainees.
Related JoVE Video
Novel algorithms for improved pattern recognition using the US FDA Adverse Event Network Analyzer.
Stud Health Technol Inform
PUBLISHED: 08-28-2014
Show Abstract
Hide Abstract
The medical review of adverse event reports for medical products requires the processing of "big data" stored in spontaneous reporting systems, such as the US Vaccine Adverse Event Reporting System (VAERS). VAERS data are not well suited to traditional statistical analyses so we developed the FDA Adverse Event Network Analyzer (AENA) and three novel network analysis approaches to extract information from these data. Our new approaches include a weighting scheme based on co-occurring triplets in reports, a visualization layout inspired by the islands algorithm, and a network growth methodology for the detection of outliers. We explored and verified these approaches by analysing the historical signal of Intussusception (IS) after the administration of RotaShield vaccine (RV) in 1999. We believe that our study supports the use of AENA for pattern recognition in medical product safety and other clinical data.
Related JoVE Video
Biochemical Characterization and Crystal Structures of a Fungal Family 3 ?-Glucosidase, Cel3A from Hypocrea jecorina.
J. Biol. Chem.
PUBLISHED: 08-27-2014
Show Abstract
Hide Abstract
Cellulase mixtures from Hypocrea jecorina are commonly used for the saccharification of cellulose in biotechnical applications. The most abundant ?-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the ?-linkage between two adjacent molecules in dimers and short oligomers of glucose. It has been shown that enhanced levels of HjCel3A in H. jecorina cellulase mixtures benefit the conversion of cellulose to glucose. Biochemical characterization of HjCel3A shows that the enzyme efficiently hydrolyzes (1,4)- as well as (1,2)-, (1,3)-, and (1,6)-?-d-linked disaccharides. For crystallization studies, HjCel3A was produced in both H. jecorina (HjCel3A) and Pichia pastoris (Pp-HjCel3A). Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. Here, we present x-ray structures of HjCel3A with and without glucose bound in the active site. The structures have a three-domain architecture as observed previously for other glycoside hydrolase family 3 ?-glucosidases. Both production hosts resulted in HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310). In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. The glycosylations are involved in intermolecular contacts in the structures derived from either host. Due to the different sizes of the glycosylations, the interactions result in different crystal forms for the two protein forms.
Related JoVE Video
Identifying Similar Cases in Document Networks using Cross-reference Structures.
IEEE J Biomed Health Inform
PUBLISHED: 08-07-2014
Show Abstract
Hide Abstract
Our objective was to explore the creation of document networks based on different thresholds of shared information and different clustering algorithms on those networks to identify document clusters describing similar clinical cases. We created networks from vaccine adverse event report sets using seven approaches for linking reports. We then applied three clustering algorithms (Visualization of Similarities [VOS], Louvain, k-means) to these networks and evaluated their ability to identify known clusters. The report sets included one simulated set and three sets from the Vaccine Adverse Event Reporting System; each was split into training and testing subsets. Training subsets were used to estimate parameter values for the clustering algorithms and testing subsets to evaluate clusters. We created the networks by linking reports based on shared information in the form either of individual Medical Dictionary for Regulatory Activities Preferred Terms (PTs) or of dyads, triplets, quadruplets, quintuplets and sextuplets of PTs; we created another network by weighting the single PT network connections by Lin's information theoretic approach to similarity. We then repeated this entire process using networks based on text mining output rather than structured data. We evaluated report clustering using recall, precision and f-measure. The VOS algorithm outperformed Louvain and k-means in general. The best weighting scheme appeared to be related to the complexity of the known cluster. For example, singleton weighting performed best for an intussusception cluster driven by a single PT. We observed marginal differences between the code- and textual-based clustering. In conclusion, our approach supported identification of similar nodes in a document network.
Related JoVE Video
Hexokinases and cardioprotection.
J. Mol. Cell. Cardiol.
PUBLISHED: 07-30-2014
Show Abstract
Hide Abstract
As mediators of the first enzymatic step in glucose metabolism, hexokinases (HKs) orchestrate a variety of catabolic and anabolic uses of glucose, regulate antioxidant power by generating NADPH for glutathione reduction, and modulate cell death processes by directly interacting with the voltage-dependent anion channel (VDAC), a regulatory component of the mitochondrial permeability transition pore (mPTP). Here we summarize the current state-of-knowledge about HKs and their role in protecting the heart from ischemia/reperfusion (I/R) injury, reviewing: 1) the properties of different HK isoforms and how their function is regulated by their subcellular localization; 2) how HKs modulate glucose metabolism and energy production during I/R; 3) the molecular mechanisms by which HKs influence mPTP opening and cellular injury during I/R; and 4) how different metabolic and HK profiles correlate with susceptibility to I/R injury and cardioprotective efficacy in cancer cells, neonatal hearts, and normal, hypertrophied and failing adult hearts, and how these difference may guide novel therapeutic strategies to limit I/R injury in the heart. This article is part of a Special Issue entitled 'Mitochondria'.
Related JoVE Video
Stereoselective formation of trisubstituted vinyl boronate esters by the acid-mediated elimination of ?-hydroxyboronate esters.
J. Org. Chem.
PUBLISHED: 07-14-2014
Show Abstract
Hide Abstract
The copper-catalyzed diboration of ketones followed by an acid-catalyzed elimination leads to the formation of 1,1-disubstituted and trisubstituted vinyl boronate esters with moderate to good yields and selectivity. Addition of tosic acid to the crude diboration products provides the corresponding vinyl boronate esters upon elimination. The trisubstituted vinyl boronate esters are formed as the (Z)-olefin isomer, which was established by subjecting the products to a Suzuki-Miyaura coupling reaction to obtain alkenes of known geometry.
Related JoVE Video
AKAP150-dependent cooperative TRPV4 channel gating is central to endothelium-dependent vasodilation and is disrupted in hypertension.
Sci Signal
PUBLISHED: 07-10-2014
Show Abstract
Hide Abstract
Endothelial cell dysfunction, characterized by a diminished response to endothelial cell-dependent vasodilators, is a hallmark of hypertension. TRPV4 channels play a major role in endothelial-dependent vasodilation, a function mediated by local Ca(2+) influx through clusters of functionally coupled TRPV4 channels rather than by a global increase in endothelial cell Ca(2+). We showed that stimulation of muscarinic acetylcholine receptors on endothelial cells of mouse arteries exclusively activated TRPV4 channels that were localized at myoendothelial projections (MEPs), specialized regions of endothelial cells that contact smooth muscle cells. Muscarinic receptor-mediated activation of TRPV4 depended on protein kinase C (PKC) and the PKC-anchoring protein AKAP150, which was concentrated at MEPs. Cooperative opening of clustered TRPV4 channels specifically amplified Ca(2+) influx at MEPs. Cooperativity of TRPV4 channels at non-MEP sites was much lower, and cooperativity at MEPs was greatly reduced by chelation of intracellular Ca(2+) or AKAP150 knockout, suggesting that Ca(2+) entering through adjacent channels underlies the AKAP150-dependent potentiation of TRPV4 activity. In a mouse model of angiotensin II-induced hypertension, MEP localization of AKAP150 was disrupted, muscarinic receptor stimulation did not activate TRPV4 channels, cooperativity among TRPV4 channels at MEPs was weaker, and vasodilation in response to muscarinic receptor stimulation was reduced. Thus, endothelial-dependent dilation of resistance arteries is enabled by MEP-localized AKAP150, which ensures the proximity of PKC to TRPV4 channels and the coupled channel gating necessary for efficient communication from endothelial to smooth muscle cells in arteries. Disruption of this molecular assembly may contribute to altered blood flow in hypertension.
Related JoVE Video
Reproducibility of Complement 4d deposition by immunofluorescence and immunohistochemistry in lung allograft biopsies.
J. Heart Lung Transplant.
PUBLISHED: 06-18-2014
Show Abstract
Hide Abstract
The significance of Complement 4d (C4d) deposition in the diagnosis of antibody-mediated rejection (AMR) in lung allografts is controversial. A potential cause may be the problematic reproducibility. We studied the reproducibility of C4d by immunofluorescence (IF) and immunohistochemistry (IHC) in lung allograft transbronchial biopsies (TBBx), correlated C4d by IF with IHC, and compared the results with clinical findings.
Related JoVE Video
Drinking games participation among female students at a regional Australian university.
J Ethn Subst Abuse
PUBLISHED: 05-24-2014
Show Abstract
Hide Abstract
Participation in drinking games (DGs) by university students is often associated with heavy drinking and negative social and health impacts. Although research in Australia indicates that university students tend to drink at risky levels, there is paucity of literature on DGs among students, especially those residing at regional universities. This research examined drinking among female college students of white background. Eighteen female students participated in face-to-face in-depth interviews to describe their DG experiences. Most women played DGs for social and monetary reasons, with many drinking high volumes of alcohol during the game. Excessive drinking was linked with the type of beverage consumed. Despite knowing the health risks associated with DGs, there was a strong social imperative for these young women to play these games. Research and public health initiatives to better understand and address problematic drinking activities in rural and regional Australia have tended to ignore women and the dominant white populations whose heavy drinking has been largely restricted to private spheres.
Related JoVE Video
Increased recombinant activated factor VII use and need for surgical reexploration following a switch from aprotinin to epsilon-aminocaproic acid in infant cardiac surgery.
J Clin Anesth
PUBLISHED: 05-05-2014
Show Abstract
Hide Abstract
To evaluate whether conversion from aprotinin to epsilon-aminocaproic acid (EACA) during infant cardiac surgery was associated with increased perioperative bleeding.
Related JoVE Video
Local control of TRPV4 channels by AKAP150-targeted PKC in arterial smooth muscle.
J. Gen. Physiol.
PUBLISHED: 04-30-2014
Show Abstract
Hide Abstract
Transient receptor potential vanilloid 4 (TRPV4) channels are Ca(2+)-permeable, nonselective cation channels expressed in multiple tissues, including smooth muscle. Although TRPV4 channels play a key role in regulating vascular tone, the mechanisms controlling Ca(2+) influx through these channels in arterial myocytes are poorly understood. Here, we tested the hypothesis that in arterial myocytes the anchoring protein AKAP150 and protein kinase C (PKC) play a critical role in the regulation of TRPV4 channels during angiotensin II (AngII) signaling. Super-resolution imaging revealed that TRPV4 channels are gathered into puncta of variable sizes along the sarcolemma of arterial myocytes. Recordings of Ca(2+) entry via single TRPV4 channels ("TRPV4 sparklets") suggested that basal TRPV4 sparklet activity was low. However, Ca(2+) entry during elementary TRPV4 sparklets was ? 100-fold greater than that during L-type CaV1.2 channel sparklets. Application of the TRPV4 channel agonist GSK1016790A or the vasoconstrictor AngII increased the activity of TRPV4 sparklets in specific regions of the cells. PKC and AKAP150 were required for AngII-induced increases in TRPV4 sparklet activity. AKAP150 and TRPV4 channel interactions were dynamic; activation of AngII signaling increased the proximity of AKAP150 and TRPV4 puncta in arterial myocytes. Furthermore, local stimulation of diacylglycerol and PKC signaling by laser activation of a light-sensitive Gq-coupled receptor (opto-?1AR) resulted in TRPV4-mediated Ca(2+) influx. We propose that AKAP150, PKC, and TRPV4 channels form dynamic subcellular signaling domains that control Ca(2+) influx into arterial myocytes.
Related JoVE Video
Old drug new use--amoxapine and its metabolites as potent bacterial ?-glucuronidase inhibitors for alleviating cancer drug toxicity.
Clin. Cancer Res.
PUBLISHED: 04-29-2014
Show Abstract
Hide Abstract
Irinotecan (CPT-11) induced diarrhea occurs frequently in patients with cancer and limits its usage. Bacteria ?-glucuronidase (GUS) enzymes in intestines convert the nontoxic metabolite of CPT-11, SN-38G, to toxic SN-38, and finally lead to damage of intestinal epithelial cells and diarrhea. We previously reported amoxapine as a potent GUS inhibitor in vitro. To further understand the molecular mechanism of amoxapine and its potential for treatment of CPT-11-induced diarrhea, we studied the binding modes of amoxapine and its metabolites by docking and molecular dynamics simulation, and tested the in vivo efficacy on mice in combination with CPT-11.
Related JoVE Video
Droperidol Transiently Prolongs the QT Interval in Children Undergoing Single Ventricle Palliation.
Pediatr Cardiol
PUBLISHED: 04-26-2014
Show Abstract
Hide Abstract
Historically, droperidol was commonly used for postoperative sedation of critically ill children. A FDA black box warning regarding its arrhythmogenic potential greatly reduced its use. We hypothesized that administration of neuroleptic dose droperidol during volatile anesthesia would transiently prolong the corrected QT interval (QTc) in patients undergoing single ventricle palliation. As part of a prospective study in children undergoing stage 2 or 3 single ventricle palliation, we recorded electrocardiograms preoperatively, after induction of volatile anesthesia, immediately after completion of 30 min intravenous infusion of 75 mcg/kg droperidol, and shortly after arrival in the cardiac intensive care unit. Mean absolute QT intervals and heart rate data were analyzed in a blinded fashion and the longest QT interval was determined. QT intervals were corrected for heart rate (QTc) with the Bazett and Friderici formulae. Any perioperative arrhythmias were recorded. Complete data were available for 62 patients. Volatile anesthesia was associated with significant prolongation of the QTc interval. Administration of droperidol after cardiopulmonary bypass was associated with further significant QTc prolongation. All QTc changes were transient and the postoperative QTc, while still prolonged relative to baseline, was significantly shorter than the QTc immediately postdroperidol. No episodes of Torsades de Pointes (TdP) or ventricular arrhythmias were observed. The administration of a neuroleptic dose of droperidol during volatile anesthesia in patients undergoing single ventricle palliation was associated with a significant prolongation of QTc, which was transient and did not result in TdP or other ventricular arrhythmias in our study population.
Related JoVE Video
Bosutinib reduces the efficacy of Dasatinib in triple-negative breast cancer cell lines.
Anticancer Res.
PUBLISHED: 04-03-2014
Show Abstract
Hide Abstract
Triple-negative breast cancer (TNBC) is an aggressive sub-type of breast cancer. Dasatinib and bosutinib are FDA-approved Src/Abl kinase inhibitor drugs. Dasatinib potently inhibits the proliferation of many TNBC cell lines.
Related JoVE Video
AKAP-anchored PKA maintains neuronal L-type calcium channel activity and NFAT transcriptional signaling.
Cell Rep
PUBLISHED: 03-18-2014
Show Abstract
Hide Abstract
L-type voltage-gated Ca2+ channels (LTCC) couple neuronal excitation to gene transcription. LTCC activity is elevated by the cyclic AMP (cAMP)-dependent protein kinase (PKA) and depressed by the Ca2+-dependent phosphatase calcineurin (CaN), and both enzymes are localized to the channel by A-kinase anchoring protein 79/150 (AKAP79/150). AKAP79/150 anchoring of CaN also promotes LTCC activation of transcription through dephosphorylation of the nuclear factor of activated T cells (NFAT). We report here that the basal activity of AKAP79/150-anchored PKA maintains neuronal LTCC coupling to CaN-NFAT signaling by preserving LTCC phosphorylation in opposition to anchored CaN. Genetic disruption of AKAP-PKA anchoring promoted redistribution of the kinase out of postsynaptic dendritic spines, profound decreases in LTCC phosphorylation and Ca2+ influx, and impaired NFAT movement to the nucleus and activation of transcription. Thus, LTCC-NFAT transcriptional signaling in neurons requires precise organization and balancing of PKA and CaN activities in the channel nanoenvironment, which is only made possible by AKAP79/150 scaffolding.
Related JoVE Video
Replication and exploratory analysis of 24 candidate risk polymorphisms for neural tube defects.
BMC Med. Genet.
PUBLISHED: 03-12-2014
Show Abstract
Hide Abstract
BackgroundNeural tube defects (NTDs), which are among the most common congenital malformations, are influenced by environmental and genetic factors. Low maternal folate is the strongest known contributing factor, making variants in genes in the folate metabolic pathway attractive candidates for NTD risk. Multiple studies have identified nominally significant allelic associations with NTDs. We tested whether associations detected in a large Irish cohort could be replicated in an independent population.MethodsReplication tests of 24 nominally significant NTD associations were performed in racially/ethnically matched populations. Family-based tests of fifteen nominally significant single nucleotide polymorphisms (SNPs) were repeated in a cohort of NTD trios (530 cases and their parents) from the United Kingdom, and case¿control tests of nine nominally significant SNPs were repeated in a cohort (190 cases, 941 controls) from New York State (NYS). Secondary hypotheses involved evaluating the latter set of nine SNPs for NTD association using alternate case¿control models and NTD groupings in white, African American and Hispanic cohorts from NYS.ResultsOf the 24 SNPs tested for replication, ADA rs452159 and MTR rs10925260 were significantly associated with isolated NTDs. Of the secondary tests performed, ARID1A rs11247593 was associated with NTDs in whites, and ALDH1A2 rs7169289 was associated with isolated NTDs in African Americans.ConclusionsWe report a number of associations between SNP genotypes and neural tube defects. These associations were nominally significant before correction for multiple hypothesis testing. These corrections are highly conservative for association studies of untested hypotheses, and may be too conservative for replication studies. We therefore believe the true effect of these four nominally significant SNPs on NTD risk will be more definitively determined by further study in other populations, and eventual meta-analysis.
Related JoVE Video
Testing and screening for chlamydia in general practice: a cross-sectional analysis.
Aust N Z J Public Health
PUBLISHED: 03-01-2014
Show Abstract
Hide Abstract
Objectives: Chlamydia screening is widely advocated. General practice registrars are an important stage of clinical behaviour development. This study aimed to determine rates of, and factors associated with, registrars' chlamydia testing including asymptomatic screening. Methods: A cross-sectional analysis of data from Registrars Clinical Encounters in Training (ReCEnT), a cohort study of registrars' consultations. Registrars record details of 60 consecutive consultations in each GP-term of training. Outcome factors were chlamydia testing, asymptomatic screening and doctor-initiated screening. Results: Testing occurred in 2.5% of 29,112 consultations (398 registrars) and in 5.8% of patients aged 15-25. Asymptomatic screening comprised 47.5% of chlamydia tests, and 55.6% of screening tests were doctor-initiated. Chlamydia testing was associated with female gender of doctor and patient, younger patient age, and patients new to doctor or practice. Asymptomatic screening was associated with practices where patients incur no fees, and in patients new to doctor or practice. Screening of female patients was more often doctor-initiated. Conclusions: GP registrars screen for chlamydia disproportionately in younger females and new patients. Implications: Our findings highlight potential opportunities to improve uptake of screening for chlamydia, including targeted education and training for registrars, campaigns targeting male patients, and addressing financial barriers to accessing screening services.
Related JoVE Video
Prevention of retinal detachment in Stickler syndrome: the Cambridge prophylactic cryotherapy protocol.
Ophthalmology
PUBLISHED: 02-14-2014
Show Abstract
Hide Abstract
The Stickler syndromes are the most common causes of inherited and childhood retinal detachment; however, no consensus exists regarding the effectiveness of prophylactic intervention. We evaluate the long-term safety and efficacy of the Cambridge prophylactic cryotherapy protocol, a standardized retinal prophylactic treatment developed to prevent retinal detachment arising from giant retinal tears in type 1 Stickler syndrome.
Related JoVE Video
Dropwise condensation of low surface tension fluids on omniphobic surfaces.
Sci Rep
PUBLISHED: 02-04-2014
Show Abstract
Hide Abstract
Compared to the significant body of work devoted to surface engineering for promoting dropwise condensation heat transfer of steam, much less attention has been dedicated to fluids with lower interfacial tension. A vast array of low-surface tension fluids such as hydrocarbons, cryogens, and fluorinated refrigerants are used in a number of industrial applications, and the development of passive means for increasing their condensation heat transfer coefficients has potential for significant efficiency enhancements. Here we investigate condensation behavior of a variety of liquids with surface tensions in the range of 12 to 28?mN/m on three types of omniphobic surfaces: smooth oleophobic, re-entrant superomniphobic, and lubricant-impregnated surfaces. We demonstrate that although smooth oleophobic and lubricant-impregnated surfaces can promote dropwise condensation of the majority of these fluids, re-entrant omniphobic surfaces became flooded and reverted to filmwise condensation. We also demonstrate that on the lubricant-impregnated surfaces, the choice of lubricant and underlying surface texture play a crucial role in stabilizing the lubricant and reducing pinning of the condensate. With properly engineered surfaces to promote dropwise condensation of low-surface tension fluids, we demonstrate a four to eight-fold improvement in the heat transfer coefficient.
Related JoVE Video
Endoscopic ultrasound in patients with normal liver blood tests and unexplained dilatation of common bile duct and or pancreatic duct.
Scand. J. Gastroenterol.
PUBLISHED: 01-28-2014
Show Abstract
Hide Abstract
To determine the yield of endoscopic ultrasound (EUS) in the investigation of patients with normal liver function tests (LFTs) and unexplained dilatation of common bile duct (CBD) and/or pancreatic duct (PD), following CT and/or magnetic resonance cholangiopancreatography.
Related JoVE Video
Is low iron status a risk factor for neural tube defects?
Birth Defects Res. Part A Clin. Mol. Teratol.
PUBLISHED: 01-08-2014
Show Abstract
Hide Abstract
Folic acid supplements can protect against neural tube defects (NTDs). Low folate and low vitamin B12 status may be maternal risk factors for having an NTD affected pregnancy. However, not all NTDs are preventable by having an adequate folate/ B12 status and other potentially modifiable factors may be involved. Folate and vitamin B12 status have important links to iron metabolism. Animal studies support an association between poor iron status and NTDs, but human data are scarce. We examined the relevance of low iron status in a nested NTD case-control study of women within a pregnant population-based cohort.
Related JoVE Video
Ankyrin-B regulates Cav2.1 and Cav2.2 channel expression and targeting.
J. Biol. Chem.
PUBLISHED: 01-06-2014
Show Abstract
Hide Abstract
N-type and P/Q-type calcium channels are documented players in the regulation of synaptic function; however, the mechanisms underlying their expression and cellular targeting are poorly understood. Ankyrin polypeptides are essential for normal integral membrane protein expression in a number of cell types, including neurons, cardiomyocytes, epithelia, secretory cells, and erythrocytes. Ankyrin dysfunction has been linked to defects in integral protein expression, abnormal cellular function, and disease. Here, we demonstrate that ankyrin-B associates with Cav2.1 and Cav2.2 in cortex, cerebellum, and brain stem. Additionally, using in vitro and in vivo techniques, we demonstrate that ankyrin-B, via its membrane-binding domain, associates with a highly conserved motif in the DII/III loop domain of Cav2.1 and Cav2.2. Further, we demonstrate that this domain is necessary for proper targeting of Cav2.1 and Cav2.2 in a heterologous system. Finally, we demonstrate that mutation of a single conserved tyrosine residue in the ankyrin-binding motif of both Cav2.1 (Y797E) and Cav2.2 (Y788E) results in loss of association with ankyrin-B in vitro and in vivo. Collectively, our findings identify an interaction between ankyrin-B and both Cav2.1 and Cav2.2 at the amino acid level that is necessary for proper Cav2.1 and Cav2.2 targeting in vivo.
Related JoVE Video
AKAP150 Contributes to Enhanced Vascular Tone by Facilitating BKCa Channel Remodeling in Hyperglycemia and Diabetes.
Circ. Res.
PUBLISHED: 12-09-2013
Show Abstract
Hide Abstract
Increased contractility of arterial myocytes and enhanced vascular tone during hyperglycemia and diabetes may arise from impaired large conductance Ca(2+)-activated K(+) (BKCa) channel function. The scaffolding protein AKAP150 is a key regulator of calcineurin (CaN), a phosphatase known to modulate expression of the regulatory BKCa ?1 subunit. Whether AKAP150 mediates BKCa channel suppression during hyperglycemia and diabetes is unknown.
Related JoVE Video
Intrinsic disorder within an AKAP-protein kinase A complex guides local substrate phosphorylation.
Elife
PUBLISHED: 11-07-2013
Show Abstract
Hide Abstract
Anchoring proteins sequester kinases with their substrates to locally disseminate intracellular signals and avert indiscriminate transmission of these responses throughout the cell. Mechanistic understanding of this process is hampered by limited structural information on these macromolecular complexes. A-kinase anchoring proteins (AKAPs) spatially constrain phosphorylation by cAMP-dependent protein kinases (PKA). Electron microscopy and three-dimensional reconstructions of type-II PKA-AKAP18? complexes reveal hetero-pentameric assemblies that adopt a range of flexible tripartite configurations. Intrinsically disordered regions within each PKA regulatory subunit impart the molecular plasticity that affords an ?16 nanometer radius of motion to the associated catalytic subunits. Manipulating flexibility within the PKA holoenzyme augmented basal and cAMP responsive phosphorylation of AKAP-associated substrates. Cell-based analyses suggest that the catalytic subunit remains within type-II PKA-AKAP18? complexes upon cAMP elevation. We propose that the dynamic movement of kinase sub-structures, in concert with the static AKAP-regulatory subunit interface, generates a solid-state signaling microenvironment for substrate phosphorylation. DOI: http://dx.doi.org/10.7554/eLife.01319.001.
Related JoVE Video
Local cAMP signaling in disease at a glance.
J. Cell. Sci.
PUBLISHED: 10-15-2013
Show Abstract
Hide Abstract
The second messenger cyclic AMP (cAMP) operates in discrete subcellular regions within which proteins that synthesize, break down or respond to the second messenger are precisely organized. A burgeoning knowledge of compartmentalized cAMP signaling is revealing how the local control of signaling enzyme activity impacts upon disease. The aim of this Cell Science at a Glance article and the accompanying poster is to highlight how misregulation of local cyclic AMP signaling can have pathophysiological consequences. We first introduce the core molecular machinery for cAMP signaling, which includes the cAMP-dependent protein kinase (PKA), and then consider the role of A-kinase anchoring proteins (AKAPs) in coordinating different cAMP-responsive proteins. The latter sections illustrate the emerging role of local cAMP signaling in four disease areas: cataracts, cancer, diabetes and cardiovascular diseases.
Related JoVE Video
Near-infrared spectroscopy: exposing the dark (venous) side of the circulation.
Paediatr Anaesth
PUBLISHED: 10-05-2013
Show Abstract
Hide Abstract
The safety of anesthesia has improved greatly in the past three decades. Standard perioperative monitoring, including pulse oximetry, has practically eliminated unrecognized arterial hypoxia as a cause for perioperative injury. However, most anesthesia-related cardiac arrests in children are now cardiovascular in origin, and standard monitoring is unable to detect many circulatory abnormalities. Near-infrared spectroscopy provides noninvasive continuous access to the venous side of regional circulations that can approximate organ-specific and global measures to facilitate the detection of circulatory abnormalities and drive goal-directed interventions to reduce end-organ ischemic injury.
Related JoVE Video
AKAP signaling complexes: pointing towards the next generation of therapeutic targets?
Trends Pharmacol. Sci.
PUBLISHED: 09-03-2013
Show Abstract
Hide Abstract
A-kinase anchoring proteins (AKAPs) streamline signal transduction by localizing signaling enzymes with their substrates. Great strides have been made in elucidating the role of these macromolecular signaling complexes as new binding partners and novel AKAPs are continually being uncovered. The mechanics and dynamics of these multi-enzyme assemblies suggest that AKAP complexes are viable targets for therapeutic intervention. This review will highlight recent advances in AKAP research focusing on local signaling events that are perturbed in disease.
Related JoVE Video
Alternative splicing modifies the effect of mutations in COL11A1 and results in recessive type 2 Stickler syndrome with profound hearing loss.
J. Med. Genet.
PUBLISHED: 08-06-2013
Show Abstract
Hide Abstract
Stickler syndromes types 1, 2 and 3 are usually dominant disorders caused by mutations in the genes COL2A1, COL11A1 and COL11A2 that encode the fibrillar collagens types II and XI present in cartilage and vitreous. Rare recessive forms of Stickler syndrome exist that are due to mutations in genes encoding type IX collagen (COL9A1 type 4 Stickler syndrome and COL9A2 type 5 Stickler syndrome). Recently, recessive mutations in the COL11A1 gene have been demonstrated to result in fibrochondrogenesis, a much more severe skeletal dysplasia, which is often lethal. Here we demonstrate that some mutations in COL11A1 are recessive, modified by alternative splicing and result in type 2 Stickler syndrome rather than fibrochondrogenesis.
Related JoVE Video
Triclosan exposure increases triclosan resistance and influences taxonomic composition of benthic bacterial communities.
Environ. Sci. Technol.
PUBLISHED: 07-25-2013
Show Abstract
Hide Abstract
Triclosan (TCS) is a broad-spectrum antimicrobial compound that is incorporated into numerous consumer products. TCS has been detected in aquatic ecosystems across the U.S., raising concern about its potential ecological effects. We conducted a field survey and an artificial stream experiment to assess effects of TCS on benthic bacterial communities. Field sampling indicated that TCS concentrations in stream sediments increased with degree of urbanization. There was significant correlation between sediment TCS concentration and the proportion of cultivable benthic bacteria that were resistant to TCS, demonstrating that the levels of TCS present in these streams was affecting the native communities. An artificial stream experiment confirmed that TCS exposure could trigger increases in TCS resistance within cultivable benthic bacteria, and pyrosequencing analysis indicated that TCS resulted in decreased benthic bacterial diversity and shifts in bacterial community composition. One notable change was a 6-fold increase in the relative abundance of cyanobacterial sequences and a dramatic die-off of algae within the artificial streams. Selection of cyanobacteria over algae could have significant implications for higher trophic levels within streams. Finally, there were no observed effects of TCS on bacterial abundance or respiration rates, suggesting that bacterial density and function were highly resilient to TCS exposure.
Related JoVE Video
Scaffolding proteins: not such innocent bystanders.
Curr. Biol.
PUBLISHED: 06-22-2013
Show Abstract
Hide Abstract
Sequential transfer of information from one enzyme to the next within the confines of a protein kinase scaffold enhances signal transduction. Though frequently considered to be inert organizational elements, two recent reports implicate kinase-scaffolding proteins as active participants in signal relay.
Related JoVE Video
Direct-acting antiviral-associated dermatitis during chronic hepatitis C virus treatment.
Am J Clin Dermatol
PUBLISHED: 06-19-2013
Show Abstract
Hide Abstract
Telaprevir and boceprevir are novel protease inhibitors recently approved for treatment of chronic hepatitis C virus infection, and have gained widespread use. Skin rash has been reported frequently in patients treated with telaprevir, but less commonly with boceprevir. Despite a high incidence in clinical trials, the telaprevir-related eruption has not been fully described in the literature. We describe six patients treated with telaprevir and three treated with boceprevir who developed skin rash related to the antiviral medication. Four patients treated with telaprevir developed laboratory abnormalities and/or systemic symptoms and five required discontinuation of their antiviral medication because of these adverse effects, including two patients who fit criteria for drug reaction with eosinophilia and systemic symptoms (DRESS). Patients with boceprevir-related rash had a milder course and none required discontinuation of the medication. This report confirms that cutaneous adverse effects from telaprevir and boceprevir are common. Patients treated with telaprevir may have a more severe course and more frequently require discontinuation of their antiviral therapy due to extensive rash or laboratory abnormalities. Dermatologists must be aware of these cutaneous adverse effects, as early intervention with topical corticosteroids and antihistamines may minimize the severity of the eruption and allow patients to complete antiviral therapy.
Related JoVE Video
Impact of continuing folic acid after the first trimester of pregnancy: findings of a randomized trial of Folic Acid Supplementation in the Second and Third Trimesters.
Am. J. Clin. Nutr.
PUBLISHED: 05-29-2013
Show Abstract
Hide Abstract
Supplementation with folic acid (FA) is recommended worldwide before and during early pregnancy because of its proven effect in preventing neural tube defects, but the role of FA after the 12th gestational week (GW) is much less clear.
Related JoVE Video
E-health: potential benefits and challenges in providing and accessing sexual health services.
BMC Public Health
PUBLISHED: 05-04-2013
Show Abstract
Hide Abstract
E-health has become a burgeoning field in which health professionals and health consumers create and seek information. E-health refers to internet-based health care and information delivery and seeks to improve health service locally, regionally and worldwide. E-sexual health presents new opportunities to provide online sexual health services irrespective of gender, age, sexual orientation and location.
Related JoVE Video
WAVE1 mediates suppression of phagocytosis by phospholipid-derived DAMPs.
J. Clin. Invest.
PUBLISHED: 05-02-2013
Show Abstract
Hide Abstract
Clearance of invading pathogens is essential to preventing overwhelming inflammation and sepsis that are symptomatic of bacterial peritonitis. Macrophages participate in this innate immune response by engulfing and digesting pathogens, a process called phagocytosis. Oxidized phospholipids (OxPL) are danger-associated molecular patterns (DAMPs) generated in response to infection that can prevent the phagocytic clearance of bacteria. We investigated the mechanism underlying OxPL action in macrophages. Exposure to OxPL induced alterations in actin polymerization, resulting in spreading of peritoneal macrophages and diminished uptake of E. coli. Pharmacological and cell-based studies showed that an anchored pool of PKA mediates the effects of OxPL. Gene silencing approaches identified the A-kinase anchoring protein (AKAP) WAVE1 as an effector of OxPL action in vitro. Chimeric Wave1(-/-) mice survived significantly longer after infection with E. coli and OxPL treatment in vivo. Moreover, we found that endogenously generated OxPL in human peritoneal dialysis fluid from end-stage renal failure patients inhibited phagocytosis via WAVE1. Collectively, these data uncover an unanticipated role for WAVE1 as a critical modulator of the innate immune response to severe bacterial infections.
Related JoVE Video
Inhibition of hepatobiliary transporters by a novel kinase inhibitor contributes to hepatotoxicity in beagle dogs.
Drug Metab Lett
PUBLISHED: 04-23-2013
Show Abstract
Hide Abstract
PF-022 (1) is a novel polycyclic benzothiophene kinase inhibitor selective for mitogen-activated protein kinase-activated protein kinase 2 (MK2). Compound 1 emerged as an inhibitor bearing submicromolar potency against MK2 (IC50 5 nM) and demonstrated projected human pharmacokinetics sufficient for oral dosing. However, following a single, oral administration of 1 to beagle dogs, animals experienced an acute liver injury characterized by increases in biomarkers associated with hepatotoxicity; particularly noteworthy was the reversible elevation in bile salts and total bilirubin. Accompanying this observation was an ADME appraisal which included hepatic bioactivation of 1 in multiple species and the in vitro inhibition of P-glycoprotein (P-gp; IC50 21 ?M). Simply attenuating the bioactivation via structural modification proved ineffective in improving the in vivo tolerability of this polycyclic scaffold. Hence, disruption of hepatobiliary transporters by the compound series was hypothesized as the likely mechanism contributing to the acute hepatotoxicity. Indeed, closer in vitro examination employing transporter gene overexpressing MDCK cell lines and membrane vesicles revealed potent compound-dependent inhibition of human multi-drug resistance-associated protein 2 (MRP2/ABCC2; IC50 38 ?M) and bile salt export pump (BSEP/ABCB11; IC50 10 ?M), two crucial hepatobiliary transport proteins accountable for bilirubin and bile salt homeostasis, respectively. Subsequent introduction of pKa-altering modifications to a second generation compound PF029 proved successful in reducing its affinity for these key efflux transporters (MRP2 IC50 >80 ?M; BSEP IC50 > 70 ?M; P-gp > 90 ?M), consequently mitigating this overt organ toxicity in dogs.
Related JoVE Video
Blood pressure in treated hypertensive individuals with the MTHFR 677TT genotype is responsive to intervention with riboflavin: findings of a targeted randomized trial.
Hypertension
PUBLISHED: 04-22-2013
Show Abstract
Hide Abstract
Intervention with riboflavin was recently shown to produce genotype-specific lowering of blood pressure (BP) in patients with premature cardiovascular disease homozygous for the 677C?T polymorphism (TT genotype) in the gene encoding the enzyme methylenetetrahydrofolate reductase (MTHFR). Whether this effect is confined to patients with high-risk cardiovascular disease is unknown. The aim of this randomized trial, therefore, was to investigate the responsiveness of BP to riboflavin supplementation in hypertensive individuals with the TT genotype but without overt cardiovascular disease. From an available sample of 1427 patients with hypertension, we identified 157 with the MTHFR 677TT genotype, 91 of whom agreed to participate in the trial. Participants were stratified by systolic BP and randomized to receive placebo or riboflavin (1.6 mg/d) for 16 weeks. At baseline, despite being prescribed multiple classes of antihypertensive drugs, >60% of participants with this genotype had failed to reach goal BP (?140/90 mm Hg). A significant improvement in the biomarker status of riboflavin was observed in response to intervention (P<0.001). Correspondingly, an overall treatment effect of 5.6±2.6 mm Hg (P=0.033) in systolic BP was observed, with pre- and postintervention values of 141.8±2.9 and 137.1±3.0 mm Hg (treatment group) and 143.5±3.0 and 144.3±3.1 mm Hg (placebo group), whereas the treatment effect in diastolic BP was not significant (P=0.291). In conclusion, these results show that riboflavin supplementation targeted at hypertensive individuals with the MTHFR 677TT genotype can decrease BP more effectively than treatment with current antihypertensive drugs only and indicate the potential for a personalized approach to the management of hypertension in this genetically at-risk group. Clinical Trial Registration- URL: http://www.clinicaltrials.gov. Unique identifier: ISRCTN23620802.
Related JoVE Video
New pleasures and old dangers: reinventing male sex work.
J Sex Res
PUBLISHED: 03-14-2013
Show Abstract
Hide Abstract
Understandings of male sex workers (MSWs) shift with technological, conceptual, and social changes. Research has historically constructed MSWs as psychologically unstable, desperate, or destitute victims and their clients as socially deviant perverts. These perceptions, however, are no longer supported by contemporary research and changing societal perceptions of the sex industry, challenging how we understand and describe "escorts." The changing understandings of sexuality and the increasing power of the Internet are both important forces behind recent changes in the structure and organization of MSWs. The growth in the visibility and reach of escorts has created opportunities to form an occupational account of MSWs that better accounts for the dynamic and diverse nature of the MSW experience in the early 21st century. Recent changes in the structure and organization of male sex work have provided visibility to the increasingly diverse geographical distribution of MSW, the commodification of race and racialized desire, new populations of heterosexual men and women as clients, and the successful dissemination of safer sexual messages to MSWs through online channels. This article provides a broad overview of the literature on MSWs, concentrating its focus on studies that have emerged over the past 20 years and identifying areas for future research.
Related JoVE Video
Effects of postchallenge administration of ST-246 on dissemination of IHD-J-Luc vaccinia virus in normal mice and in immune-deficient mice reconstituted with T cells.
J. Virol.
PUBLISHED: 03-06-2013
Show Abstract
Hide Abstract
Whole-body bioimaging was used to study dissemination of vaccinia virus (VACV) in normal and in immune deficient (nu(-)/nu(-)) mice protected from lethality by postchallenge administration of ST-246. Total fluxes were recorded in the liver, spleen, lungs, and nasal cavities of live mice after intranasal infection with a recombinant IHD-J-Luc VACV expressing luciferase. Areas under the flux curve were calculated for individual mice to assess viral loads. Treatment for 2 to 5 days of normal BALB/c mice with ST-246 at 100 mg/kg starting 24 h postchallenge conferred 100% protection and reduced viral loads in four organs compared to control mice. Mice also survived after 5 days of treatment with ST-246 at 30 mg/kg, and yet the viral loads and poxes were higher in these mice compared to 100-mg/kg treatment group. Nude mice were not protected by ST-246 alone or by 10 million adoptively transferred T cells. In contrast, nude mice that received T cells and 7-day treatment with ST-246 survived infection and exhibited reduced viral loads compared to nonreconstituted and ST-246-treated mice after ST-246 was stopped. Similar protection of nude mice was achieved using adoptively transferred 1.0 and 0.1 million, but not 0.01 million, purified T cells or CD4(+) or CD8(+) T cells in conjunction with ST-246 treatment. These data suggest that ST-246 protects immunocompetent mice from lethality and reduces viral dissemination in internal organs and poxvirus lesions. Furthermore, immune-deficient animals with partial T cell reconstitution can control virus replication after a course of ST-246 and survive lethal vaccinia virus challenge.
Related JoVE Video
Prevalence, clinical and virologic outcomes of hepatitis B virus co-infection in HIV-1 positive Kenyan women on antiretroviral therapy.
PLoS ONE
PUBLISHED: 02-13-2013
Show Abstract
Hide Abstract
Sub-Saharan Africa carries a high burden of co-infection with HIV-1 and hepatitis B virus (HBV). In this region, individuals with HIV-1/HBV co-infection on antiretroviral therapy (ART) frequently receive lamivudine as the only agent active against HBV, raising concerns for development of HBV resistance to lamivudine. We aimed to determine the prevalence, clinical, and virologic outcomes of chronic HBV infection, including HBV resistance to lamivudine, in a cohort of HIV-1 seropositive Kenyan women on long-term ART.
Related JoVE Video
Bayesian methods for design and analysis of safety trials.
Pharm Stat
PUBLISHED: 02-11-2013
Show Abstract
Hide Abstract
Safety assessment is essential throughout medical product development. There has been increased awareness of the importance of safety trials recently, in part due to recent US Food and Drug Administration guidance related to thorough assessment of cardiovascular risk in the treatment of type 2 diabetes. Bayesian methods provide great promise for improving the conduct of safety trials. In this paper, the safety subteam of the Drug Information Association Bayesian Scientific Working Group evaluates challenges associated with current methods for designing and analyzing safety trials and provides an overview of several suggested Bayesian opportunities that may increase efficiency of safety trials along with relevant case examples. Copyright © 2013 John Wiley & Sons, Ltd.
Related JoVE Video
Chemical and biological characterization of wastewater generated from hydrothermal liquefaction of Spirulina.
Environ. Sci. Technol.
PUBLISHED: 02-11-2013
Show Abstract
Hide Abstract
Hydrothermal liquefaction (HTL) is an attractive method for converting wet biomass into petroleum-like biocrude oil that can be refined to make petroleum products. This approach is advantageous for conversion of low-lipid algae, which are promising feedstocks for sustainable large-scale biofuel production. As with natural petroleum formation, the water in contact with the produced oil contains toxic compounds. The objectives of this research were to: (1) identify nitrogenous organic compounds (NOCs) in wastewater from HTL conversion of Spirulina; (2) characterize mammalian cell cytotoxicity of specific NOCs, NOC mixture, and the complete HTL wastewater (HTL-WW) matrix; and (3) investigate mitigation measures to reduce toxicity in HTL-WW. Liquid-liquid extraction and nitrogen-phosphorus detection was used in conjunction with gas chromatography-mass spectrometry (GC-MS), which detected hundreds of NOCs in HTL-WW. Reference materials for nine of the most prevalent NOCs were used to identify and quantify their concentrations in HTL-WW. Mammalian cell cytotoxicity of the nine NOCs was quantified using a Chinese hamster ovary (CHO) cell assay, and the descending rank order for cytotoxicity was 3-dimethylamino-phenol > 2,2,6,6-tetramethyl-4-piperidone > 2,6-dimethyl-3-pyridinol > 2-picoline > pyridine > 1-methyl-2-pyrrolidinone > ?-valerolactam > 2-pyrrolidinone > ?-caprolactam. The organic mixture extracted from HTL-WW expressed potent CHO cell cytotoxic activity, with a LC(50) at 7.5% of HTL-WW. Although the toxicity of HTL-WW was substantial, 30% of the toxicity was removed biologically by recycling HTL-WW back into algal cultivation. The remaining toxicity of HTL-WW was mostly eliminated by subsequent treatment with granular activated carbon.
Related JoVE Video
Arbuscular-mycorrhizal networks inhibit Eucalyptus tetrodonta seedlings in rain forest soil microcosms.
PLoS ONE
PUBLISHED: 01-24-2013
Show Abstract
Hide Abstract
Eucalyptus tetrodonta, a co-dominant tree species of tropical, northern Australian savannas, does not invade adjacent monsoon rain forest unless the forest is burnt intensely. Such facilitation by fire of seedling establishment is known as the "ashbed effect." Because the ashbed effect might involve disruption of common mycorrhizal networks, we hypothesized that in the absence of fire, intact rain forest arbuscular mycorrhizal (AM) networks inhibit E. tetrodonta seedlings. Although arbuscular mycorrhizas predominate in the rain forest, common tree species of the northern Australian savannas (including adult E. tetrodonta) host ectomycorrhizas. To test our hypothesis, we grew E. tetrodonta and Ceiba pentandra (an AM-responsive species used to confirm treatments) separately in microcosms of ambient or methyl-bromide fumigated rain forest soil with or without severing potential mycorrhizal fungus connections to an AM nurse plant, Litsea glutinosa. As expected, C. pentandra formed mycorrhizas in all treatments but had the most root colonization and grew fastest in ambient soil. E. tetrodonta seedlings also formed AM in all treatments, but severing hyphae in fumigated soil produced the least colonization and the best growth. Three of ten E. tetrodonta seedlings in ambient soil with intact network hyphae died. Because foliar chlorosis was symptomatic of iron deficiency, after 130 days we began to fertilize half the E. tetrodonta seedlings in ambient soil with an iron solution. Iron fertilization completely remedied chlorosis and stimulated leaf growth. Our microcosm results suggest that in intact rain forest, common AM networks mediate belowground competition and AM fungi may exacerbate iron deficiency, thereby enhancing resistance to E. tetrodonta invasion. Common AM networks-previously unrecognized as contributors to the ashbed effect-probably help to maintain the rain forest-savanna boundary.
Related JoVE Video
Choline supplementation and measures of choline and betaine status: a randomised, controlled trial in postmenopausal women.
Br. J. Nutr.
PUBLISHED: 12-15-2011
Show Abstract
Hide Abstract
Choline is an essential nutrient and can also be obtained by de novo synthesis via an oestrogen responsive pathway. Choline can be oxidised to the methyl donor betaine, with short-term supplementation reported to lower plasma total homocysteine (tHcy); however, the effects of longer-term choline supplementation are less clear. We investigated the effect of choline supplementation on plasma concentrations of free choline, betaine and tHcy and B-vitamin status in postmenopausal women, a group more susceptible to low choline status. We also assessed whether supplementation altered plasma lipid profiles. In this randomised, double-blinded, placebo-controlled study, forty-two healthy postmenopausal women received 1 g choline per d (as choline bitartrate), or an identical placebo supplement with their habitual diet. Fasting blood samples were collected at baseline, week 6 and week 12. Administration of choline increased median choline and betaine concentrations in plasma, with significant effects evident after 6 weeks of supplementation (P<0·001) and remaining significant at 12 weeks (P<0·001); no effect was observed on folate status or on plasma lipids. Choline supplementation induced a median (25th, 75th percentile) change in plasma tHcy concentration at week 6 of -0·9 (-1·6, 0·2) ?mol, a change which, when compared to that observed in the placebo group 0·6 (-0·4, 1·9) ?mol, approached statistical significance (P=0·058). Choline supplementation at a dose of 1 g/d significantly increases the circulating concentration of free choline, and can also significantly increase the concentration of the methyl donor, betaine, thereby potentially enhancing the betaine-homocysteine methyltransferase-mediated remethylation of tHcy.
Related JoVE Video
Folic acid consumption throughout pregnancy: differentiation between trimesters.
Ann. Nutr. Metab.
PUBLISHED: 11-25-2011
Show Abstract
Hide Abstract
Intervention trials have demonstrated conclusively that oral ingestion of extra folic acid can prevent the majority of cases of spina bifida and other neural tube defects (NTDs). Data from these studies offer conclusive evidence that the intake of 400 ?g/day folic acid provides this benefit. The big problem has remained that the neural plate closes to form the neural tube between day 21 and day 28 postconception, at which point most women do not even realize that they are pregnant. This, coupled with the fact that over half of all pregnancies are unplanned, has resulted in many studies showing that less than one-fifth of pregnancies have followed the recommendation for prevention. However, where compliance has been good or where folic acid has been added mandatorily, there has been the expected reduction in NTDs. Clarity regarding what is recommended for early pregnancy does not carry through to the second or third trimester of pregnancy where practices vary widely not only from country to country but even from clinician to clinician within particular countries.
Related JoVE Video
An entirely specific type I A-kinase anchoring protein that can sequester two molecules of protein kinase A at mitochondria.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 11-14-2011
Show Abstract
Hide Abstract
A-kinase anchoring proteins (AKAPs) tether the cAMP-dependent protein kinase (PKA) to intracellular sites where they preferentially phosphorylate target substrates. Most AKAPs exhibit nanomolar affinity for the regulatory (RII) subunit of the type II PKA holoenzyme, whereas dual-specificity anchoring proteins also bind the type I (RI) regulatory subunit of PKA with 10-100-fold lower affinity. A range of cellular, biochemical, biophysical, and genetic approaches comprehensively establish that sphingosine kinase interacting protein (SKIP) is a truly type I-specific AKAP. Mapping studies located anchoring sites between residues 925-949 and 1,140-1,175 of SKIP that bind RI with dissociation constants of 73 and 774 nM, respectively. Molecular modeling and site-directed mutagenesis approaches identify Phe 929 and Tyr 1,151 as RI-selective binding determinants in each anchoring site. SKIP complexes exist in different states of RI-occupancy as single-molecule pull-down photobleaching experiments show that 41 ± 10% of SKIP sequesters two YFP-RI dimers, whereas 59 ± 10% of the anchoring protein binds a single YFP-RI dimer. Imaging, proteomic analysis, and subcellular fractionation experiments reveal that SKIP is enriched at the inner mitochondrial membrane where it associates with a prominent PKA substrate, the coiled-coil helix protein ChChd3.
Related JoVE Video
Dynamics of nanoparticle self-assembly into superhydrophobic liquid marbles during water condensation.
ACS Nano
PUBLISHED: 11-03-2011
Show Abstract
Hide Abstract
Nanoparticles adsorbed onto the surface of a drop can fully encapsulate the liquid, creating a robust and durable soft solid with superhydrophobic characteristics referred to as a liquid marble. Artificially created liquid marbles have been studied for about a decade but are already utilized in some hair and skin care products and have numerous other potential applications. These soft solids are usually formed in small quantity by depositing and rolling a drop of liquid on a layer of hydrophobic particles but can also be made in larger quantities in an industrial mixer. In this work, we demonstrate that microscale liquid marbles can also form through self-assembly during water condensation on a superhydrophobic surface covered with a loose layer of hydrophobic nanoparticles. Using in situ environmental scanning electron microscopy and optical microscopy, we study the dynamics of liquid marble formation and evaporation as well as their interaction with condensing water droplets. We demonstrate that the self-assembly of nanoparticle films into three-dimensional liquid marbles is driven by multiple coalescence events between partially covered droplets and is aided by surface flows causing rapid nanoparticle film redistribution. We also show that droplet and liquid marble coalescence can occur due to liquid-to-liquid contact or squeezing of the two objects into each other as a result of compressive forces from surrounding droplets and marbles. Irrelevant of the mechanism, coalescence of marbles and drops can cause their rapid movement across and rolling off the edge of the surface. We also demonstrate that the liquid marbles randomly moving across the surface can be captured and immobilized by hydrophilic surface patterns.
Related JoVE Video
Biochemical and molecular characterization of secreted ?-xylosidase from Aspergillus niger.
J. Biol. Chem.
PUBLISHED: 10-27-2011
Show Abstract
Hide Abstract
?-Linked xylose is a major component of xyloglucans in the cell walls of higher plants. An ?-xylosidase (AxlA) was purified from a commercial enzyme preparation from Aspergillus niger, and the encoding gene was identified. The protein is a member of glycosyl hydrolase family 31. It was active on p-nitrophenyl-?-d-xyloside, isoprimeverose, xyloglucan heptasaccharide (XXXG), and tamarind xyloglucan. When expressed in Pichia pastoris, AxlA had activity comparable to the native enzyme on pNP?X and IP despite apparent hyperglycosylation. The pH optimum of AxlA was between 3.0 and 4.0. AxlA together with ?-glucosidase depolymerized xyloglucan heptasaccharide. A combination of AxlA, ?-glucosidase, xyloglucanase, and ?-galactosidase in the optimal proportions of 51:5:19:25 or 59:5:11:25 could completely depolymerize tamarind XG to free Glc or Xyl, respectively. To the best of our knowledge, this is the first characterization of a secreted microbial ?-xylosidase. Secreted ?-xylosidases appear to be rare in nature, being absent from other tested commercial enzyme mixtures and from the genomes of most filamentous fungi.
Related JoVE Video
B-vitamins, methylenetetrahydrofolate reductase (MTHFR) and hypertension.
Int J Vitam Nutr Res
PUBLISHED: 10-25-2011
Show Abstract
Hide Abstract
Hypertension is a leading risk factor for cardiovascular disease (CVD) and stroke. A common polymorphism in the gene encoding the enzyme methylenetetrahydrofolate reductase (MTHFR), previously identified as the main genetic determinant of elevated homocysteine concentration and also recognized as a risk factor for CVD, appears to be independently associated with hypertension. The B-vitamin riboflavin is required as a cofactor by MTHFR and recent evidence suggests it may have a role in modulating blood pressure, specifically in those with the homozygous mutant MTHFR 677 TT genotype. If studies confirm that this genetic predisposition to hypertension is correctable by low-dose riboflavin, the findings could have important implications for the management of hypertension given that the frequency of this polymorphism ranges from 3 to 32 % worldwide.
Related JoVE Video
Ribosomal biosynthesis of ?-amanitin in Galerina marginata.
Fungal Genet. Biol.
PUBLISHED: 09-09-2011
Show Abstract
Hide Abstract
Amatoxins, including ?-amanitin, are bicyclic octapeptides found in mushrooms (Agaricomycetes, Agaricales) of certain species in the genera Amanita, Galerina, Lepiota, and Conocybe. Amatoxins and the chemically similar phallotoxins are synthesized on ribosomes in Amanita bisporigera, Amanita phalloides, and Amanita ocreata. In order to determine if amatoxins are synthesized by a similar mechanism in another, distantly related mushroom, we obtained genome survey sequence data from a monokaryotic isolate of Galerinamarginata, which produces ?-amanitin. The genome of G. marginata contains two copies of the ?-amanitin gene (GmAMA1-1 and GmAMA1-2). The ?-amanitin proprotein sequences of G. marginata (35 amino acids) are highly divergent from AMA1 of A. bisporigera except for the toxin region itself (IWGIGCNP in single-letter amino acid code) and the amino acids immediately upstream (N[A/S]TRLP). G. marginata does not contain any related toxin-encoding sequences besides GmAMA1-1 and GmAMA1-2. DNA from two other ?-amanitin-producing isolates of Galerina (G. badipes and G. venenata) hybridized to GmAMA1, whereas DNA from the toxin non-producing species Galerinahybrida did not. Expression of the GmAMA1 genes was induced by growth on low carbon. RNASeq evidence indicates that both copies of GmAMA1 are expressed approximately equally. A prolyl oligopeptidase (POP) is strongly implicated in processing of the cyclic peptide toxins of A. bisporigera and Conocybe apala. G. marginata has two predicted POP genes; one, like AbPOPB of A. bisporigera, is present only in the toxin-producing isolates of Galerina and the other, like AbPOPA of A. bisporigera, is present in all species. Our results indicate that G.marginata biosynthesizes amatoxins on ribosomes by a pathway similar to Amanita species, involving a genetically encoded proprotein of 35 amino acids that is post-translationally processed by a POP. However, due to the high degree of divergence, the evolutionary relationship between AMA1 in the genera Amanita and Galerina is unclear.
Related JoVE Video
AKAP220 protein organizes signaling elements that impact cell migration.
J. Biol. Chem.
PUBLISHED: 09-02-2011
Show Abstract
Hide Abstract
Cell movement requires the coordinated reception, integration, and processing of intracellular signals. We have discovered that the protein kinase A anchoring protein AKAP220 interacts with the cytoskeletal scaffolding protein IQGAP1 to influence cell motility. AKAP220/IQGAP1 networks receive and integrate calcium and cAMP second messenger signals and position signaling enzymes near their intended substrates at leading edges of migrating cells. IQGAP1 supports calcium/calmodulin-dependent association of factors that modulate microtubule dynamics. AKAP220 suppresses GSK-3? and positions this kinase to allow recruitment of the plus-end microtubule tracking protein CLASP2. Gene silencing of AKAP220 alters the rate of microtubule polymerization and the lateral tracking of growing microtubules and retards cell migration in metastatic human cancer cells. This reveals an unappreciated role for this anchored kinase/microtubule effector protein network in the propagation of cell motility.
Related JoVE Video
AKAP2 anchors PKA with aquaporin-0 to support ocular lens transparency.
EMBO Mol Med
PUBLISHED: 08-30-2011
Show Abstract
Hide Abstract
A decline in ocular lens transparency known as cataract afflicts 90% of individuals by the age 70. Chronic deterioration of lens tissue occurs as a pathophysiological consequence of defective water and nutrient circulation through channel and transporter proteins. A key component is the aquaporin-0 (AQP0) water channel whose permeability is tightly regulated in healthy lenses. Using a variety of cellular and biochemical approaches we have discovered that products of the A-kinase anchoring protein 2 gene (AKAP2/AKAP-KL) form a stable complex with AQP0 to sequester protein kinase A (PKA) with the channel. This permits PKA phosphorylation of serine 235 within a calmodulin (CaM)-binding domain of AQP0. The additional negative charge introduced by phosphoserine 235 perturbs electrostatic interactions between AQP0 and CaM to favour water influx through the channel. In isolated mouse lenses, displacement of PKA from the AKAP2-AQP0 channel complex promotes cortical cataracts as characterized by severe opacities and cellular damage. Thus, anchored PKA modulation of AQP0 is a homeostatic mechanism that must be physically intact to preserve lens transparency.
Related JoVE Video
Widespread dispersal of Borrelia burgdorferi-infected ticks collected from songbirds across Canada.
J. Parasitol.
PUBLISHED: 08-24-2011
Show Abstract
Hide Abstract
Millions of Lyme disease vector ticks are dispersed annually by songbirds across Canada, but often overlooked as the source of infection. For clarity on vector distribution, we sampled 481 ticks (12 species and 3 undetermined ticks) from 211 songbirds (42 species/subspecies) nationwide. Using PCR, 52 (29.5%) of 176 Ixodes ticks tested were positive for the Lyme disease spirochete, Borrelia burgdorferi s.l. Immature blacklegged ticks, Ixodes scapularis , collected from infested songbirds had a B. burgdorferi infection prevalence of 36% (larvae, 48%; nymphs, 31%). Notably, Ixodes affinis is reported in Canada for the first time and, similarly, Ixodes auritulus for the initial time in the Yukon. Firsts for bird-parasitizing ticks include I. scapularis in Quebec and Saskatchewan. We provide the first records of 3 tick species cofeeding on passerines (song sparrow, Swainsons thrush). New host records reveal I. scapularis on the blackpoll warbler and Nashville warbler. We furnish the following first Canadian reports of B. burgdorferi-positive ticks: I. scapularis on chipping sparrow, house wren, indigo bunting; I. auritulus on Bewicks wren; and I. spinipalpis on a Bewicks wren and song sparrow. First records of B. burgdorferi-infected ticks on songbirds include the following: the rabbit-associated tick, Ixodes dentatus, in western Canada; I. scapularis in Quebec, Saskatchewan, northern New Brunswick, northern Ontario; and Ixodes spinipalpis (collected in British Columbia). The presence of B. burgdorferi in Ixodes larvae suggests reservoir competency in 9 passerines (Bewicks wren, common yellowthroat, dark-eyed junco, Oregon junco, red-winged blackbird, song sparrow, Swainsons thrush, swamp sparrow, and white-throated sparrow). We report transstadial transmission (larva to nymph) of B. burgdorferi in I. auritulus. Data suggest a possible 4-tick, i.e., I. angustus, I. auritulus, I. pacificus, and I. spinipalpis, enzootic cycle of B. burgdorferi on Vancouver Island, British Columbia. Our results suggest that songbirds infested with B. burgdorferi-infected ticks have the potential to start new tick populations endemic for Lyme disease. Because songbirds disperse B. burgdorferi-infected ticks outside their anticipated range, health-care providers are advised that people can contract Lyme disease locally without any history of travel.
Related JoVE Video
Characterization of polygenic resistance to powdery mildew in tomato at cytological, biochemical and gene expression level.
Mol. Plant Pathol.
PUBLISHED: 08-22-2011
Show Abstract
Hide Abstract
Extensive research in the area of plant innate immunity has increased considerably our understanding of the molecular mechanisms associated with resistance controlled by a dominant resistance gene. In contrast, little is known about the molecular basis underlying the resistance conferred by quantitative trait loci (QTLs). In this study, using the interaction of tomato (Solanum lycopersicum) with Oidium neolycopersici, we compared the cytological, biochemical and molecular mechanisms involved in both monogenic and polygenic resistances conferred by a dominant gene (Ol-1) and three QTLs (Ol-qtls), respectively. Our results showed that the three Ol-qtls jointly confer a very high level of broad-spectrum resistance and that the resistance is associated with both the hypersensitive response and papillae formation, with the hypersensitive response being prevalent. Both H(2)O(2) and callose accumulation, which are coupled with Ol-1-mediated resistance, are also associated with the resistance conferred by Ol-qtls. Further, we analysed the pathogen-induced transcript profiles of near-isogenic lines carrying the three Ol-qtls and the Ol-1 gene. Transcript profiles obtained by cDNA-amplified fragment length polymorphism analysis showed that, on fungal challenge, about 70% of the transcript-derived fragments are up-regulated in both susceptible and resistant genotypes. Most of the sequenced transcript-derived fragments showed homology to genes with functions in defence responses, suggesting that defence-responsive genes responsible for basal defence are involved in both monogenic and polygenic resistances conferred by Ol-1 and Ol-qtls, respectively. Although about 18% of the identified transcript-derived fragments are specific for either monogenic or polygenic resistance, their expression patterns need to be further verified by quantitative reverse transcriptase-polymerase chain reaction.
Related JoVE Video
HBV infection in relation to consistent condom use: a population-based study in Peru.
PLoS ONE
PUBLISHED: 08-18-2011
Show Abstract
Hide Abstract
Data on hepatitis B virus (HBV) prevalence are limited in developing countries. There is also limited information of consistent condom use efficacy for reducing HBV transmission at the population level. The study goal was to evaluate the prevalence and factors associated with HBV infection in Peru, and the relationship between anti-HBc positivity and consistent condom use.
Related JoVE Video
Genetic and functional heterogeneity of the hepatitis C virus p7 ion channel during natural chronic infection.
Virology
PUBLISHED: 07-13-2011
Show Abstract
Hide Abstract
The present study describes natural genetic heterogeneity of hepatitis C virus (HCV) p7 protein, the ion channel that plays a critical role in assembly and release of HCV, within 299 variants isolated from serum specimens of 27 chronically infected patients, 12 of whom with human immunodeficiency virus (HIV) co-infection. Liver fibrosis stage was inversely correlated with p7 synonymous substitutions (dS) (p=0.033), and indices of p7 genetic diversity were significantly higher in HIV-negative subjects compared to HIV-positive subjects (dS, p=0.005; non-synonymous substitutions (dN), p=0.002; dN/dS ratio, p=0.024; amino acid distances, p=0.007). Six p7 genes with naturally occurring unique amino acid variations were selected for in vitro study. The variants demonstrated diversified functional heterogeneity in vitro, with one variant from a subject with severe liver disease displaying hyperactive ion channel function, as well as other variants presenting altered pH-activated channel gating activities.
Related JoVE Video
IL28B genotype effects during early treatment with peginterferon and ribavirin in difficult-to-treat hepatitis C virus infection.
J. Infect. Dis.
PUBLISHED: 07-12-2011
Show Abstract
Hide Abstract
Mathematical models of hepatitis C virus (HCV) during therapy may elucidate mechanisms of action for antiviral therapy. In genome-wide association studies, IL28B gene polymorphisms are highly predictive of therapeutic clearance of HCV.
Related JoVE Video
Passive immunotherapies protect WRvFire and IHD-J-Luc vaccinia virus-infected mice from lethality by reducing viral loads in the upper respiratory tract and internal organs.
J. Virol.
PUBLISHED: 06-29-2011
Show Abstract
Hide Abstract
Whole-body bioimaging was employed to study the effects of passive immunotherapies on lethality and viral dissemination in BALB/c mice challenged with recombinant vaccinia viruses expressing luciferase. WRvFire and IHD-J-Luc vaccinia viruses induced lethality with similar times to death following intranasal infection, but WRvFire replicated at higher levels than IHD-J-Luc in the upper and lower respiratory tracts. Three types of therapies were tested: licensed human anti-vaccinia virus immunoglobulin intravenous (VIGIV); recombinant anti-vaccinia virus immunoglobulin (rVIG; Symphogen, Denmark), an investigational product containing a mixture of 26 human monoclonal antibodies (HuMAbs) against mature virion (MV) and enveloped virion (EV); and HuMAb compositions targeting subsets of MV or EV proteins. Bioluminescence recorded daily showed that pretreatment with VIGIV (30 mg) or with rVIG (100 ?g) on day -2 protected mice from death but did not prevent viral replication at the site of inoculation and dissemination to internal organs. Compositions containing HuMAbs against MV or EV proteins were protective in both infection models at 100 ?g per animal, but at 30 ?g, only anti-EV antibodies conferred protection. Importantly, the t statistic of the mean total fluxes revealed that viral loads in surviving mice were significantly reduced in at least 3 sites for 3 consecutive days (days 3 to 5) postchallenge, while significant reduction for 1 or 2 days in any individual site did not confer protection. Our data suggest that reduction of viral replication at multiple sites, including respiratory tract, spleen, and liver, as monitored by whole-body bioluminescence can be used to predict the effectiveness of passive immunotherapies in mouse models.
Related JoVE Video
Restoration of normal L-type Ca2+ channel function during Timothy syndrome by ablation of an anchoring protein.
Circ. Res.
PUBLISHED: 06-23-2011
Show Abstract
Hide Abstract
L-type Ca(2+) (Ca(V)1.2) channels shape the cardiac action potential waveform and are essential for excitation-contraction coupling in heart. A gain-of-function G406R mutation in a cytoplasmic loop of Ca(V)1.2 channels causes long QT syndrome 8 (LQT8), a disease also known as Timothy syndrome. However, the mechanisms by which this mutation enhances Ca(V)1.2-LQT8 currents and generates lethal arrhythmias are unclear.
Related JoVE Video
Methodology for imaging nano-to-microscale water condensation dynamics on complex nanostructures.
ACS Nano
PUBLISHED: 06-15-2011
Show Abstract
Hide Abstract
A better understanding of the role that nanoscale surface chemical heterogeneities and topographical features play in water droplet formation is necessary to improve design and robustness of nanostructured superhydrophobic surfaces as to make them fit for industrial applications. Lack of an imaging method capable of capturing the water condensation process on complex nanostructures with required magnification has thus far hindered experimental progress in this area. In this work, we demonstrate that by transferring a small part of a macroscale sample to a novel thermally insulated sample platform we are able to mitigate flooding and electron heating problems typically associated with environmental scanning electron microscopy of water condensation. We image condensation dynamics on individual complex particles and a superhydrophobic network of nanostructures fabricated from low thermal conductivity materials with an unobstructed 90° perspective of the surface-to-water interface with field of view as small as 1 ?m(2). We clearly observe the three-stage drop growth process and demonstrate that even during late stages of the droplet growth the nearly spherical drop remains in a partially wetting Wenzel state.
Related JoVE Video
Alkaline peroxide pretreatment of corn stover: effects of biomass, peroxide, and enzyme loading and composition on yields of glucose and xylose.
Biotechnol Biofuels
PUBLISHED: 06-09-2011
Show Abstract
Hide Abstract
Pretreatment is a critical step in the conversion of lignocellulose to fermentable sugars. Although many pretreatment processes are currently under investigation, none of them are entirely satisfactory in regard to effectiveness, cost, or environmental impact. The use of hydrogen peroxide at pH 11.5 (alkaline hydrogen peroxide (AHP)) was shown by Gould and coworkers to be an effective pretreatment of grass stovers and other plant materials in the context of animal nutrition and ethanol production. Our earlier experiments indicated that AHP performed well when compared against two other alkaline pretreatments. Here, we explored several key parameters to test the potential of AHP for further improvement relevant to lignocellulosic ethanol production.
Related JoVE Video
Mucocele of the cystic duct remnant after orthotopic liver transplant: a problem revisited.
Exp Clin Transplant
PUBLISHED: 06-09-2011
Show Abstract
Hide Abstract
Mucocele of the cystic duct remnant is an uncommon hepatobiliary complication of a liver transplant. Current practice usually involves either excising the cystic duct, or incorporating the distal end of the transected cystic duct into the suture line of the biliary anastomosis to ensure drainage. We report a patient who developed cystic duct remnant mucocele after the latter approach was adopted. We believe that this is likely related to delayed anastomotic stricturing, which prevented draining from the remnant cystic duct. We also discuss the incidence, pathology, investigations, and treatment of this condition.
Related JoVE Video
Do high blood folate concentrations exacerbate metabolic abnormalities in people with low vitamin B-12 status?
Am. J. Clin. Nutr.
PUBLISHED: 06-08-2011
Show Abstract
Hide Abstract
In elderly individuals with low serum vitamin B-12, those who have high serum folate have been reported to have greater abnormalities in the following biomarkers for vitamin B-12 deficiency: low hemoglobin and elevated total homocysteine (tHcy) and methylmalonic acid (MMA). This suggests that folate exacerbates vitamin B-12-related metabolic abnormalities.
Related JoVE Video
Discovery of cellular substrates for protein kinase A using a peptide array screening protocol.
Biochem. J.
PUBLISHED: 06-08-2011
Show Abstract
Hide Abstract
Post-translational modification of proteins is a universal form of cellular regulation. Phosphorylation on serine, threonine, tyrosine or histidine residues by protein kinases is the most widespread and versatile form of covalent modification. Resultant changes in activity, localization or stability of phosphoproteins drives cellular events. MS and bioinformatic analyses estimate that ~30% of intracellular proteins are phosphorylated at any given time. Multiple approaches have been developed to systematically define targets of protein kinases; however, it is likely that we have yet to catalogue the full complement of the phosphoproteome. The amino acids that surround a phosphoacceptor site are substrate determinants for protein kinases. For example, basophilic enzymes such as PKA (protein kinase A), protein kinase C and calmodulin-dependent kinases recognize basic side chains preceding the target serine or threonine residues. In the present paper we describe a strategy using peptide arrays and motif-specific antibodies to identify and characterize previously unrecognized substrate sequences for protein kinase A. We found that the protein kinases PKD (protein kinase D) and MARK3 [MAP (microtubule-associated protein)-regulating kinase 3] can both be phosphorylated by PKA. Furthermore, we show that the adapter protein RIL [a product of PDLIM4 (PDZ and LIM domain protein 4)] is a PKA substrate that is phosphorylated on Ser(119) inside cells and that this mode of regulation may control its ability to affect cell growth.
Related JoVE Video
Patient acceptance of universal screening for hepatitis C virus infection.
BMC Infect. Dis.
PUBLISHED: 06-06-2011
Show Abstract
Hide Abstract
In the United States, about 70% of 2.9-3.7 million people with hepatitis C (HCV) are unaware of their infection. Although universal screening might be a cost-effective way to identify infections, prevent morbidity, and reduce transmission, few efforts have been made to determine patient opinions about new approaches to screening.
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.