247 research outputs found
Identification and functional characterisation of CRK12:CYC9, a novel cyclin-dependent kinase (CDK)-cyclin complex in Trypanosoma brucei
The protozoan parasite, Trypanosoma brucei, is spread by the tsetse fly and causes trypanosomiasis in humans and animals. Both the life cycle and cell cycle of the parasite are complex. Trypanosomes have eleven cdc2-related kinases (CRKs) and ten cyclins, an unusually large number for a single celled organism. To date, relatively little is known about the function of many of the CRKs and cyclins, and only CRK3 has previously been shown to be cyclin-dependent in vivo. Here we report the identification of a previously uncharacterised CRK:cyclin complex between CRK12 and the putative transcriptional cyclin, CYC9. CRK12:CYC9 interact to form an active protein kinase complex in procyclic and bloodstream T. brucei. Both CRK12 and CYC9 are essential for the proliferation of bloodstream trypanosomes in vitro, and we show that CRK12 is also essential for survival of T. brucei in a mouse model, providing genetic validation of CRK12:CYC9 as a novel drug target for trypanosomiasis. Further, functional characterisation of CRK12 and CYC9 using RNA interference reveals roles for these proteins in endocytosis and cytokinesis, respectively
Determinants of legacy effects in pine trees â implications from an irrigation-stop experiment
Tree responses to altered water availability range from immediate (e.g. stomatal regulation) to delayed (e.g. crown size adjustment). The interplay of the different response times and processes, and their effects on long-term whole-tree performance, however, is hardly understood. Here we investigated legacy effects on structures and functions of mature Scots pine in a dry inner-Alpine Swiss valley after stopping an 11-yr lasting irrigation treatment. Measured ecophysiological time series were analysed and interpreted with a system-analytic tree model. We found that the irrigation stop led to a cascade of downregulations of physiological and morphological processes with different response times. Biophysical processes responded within days, whereas needle and shoot lengths, crown transparency, and radial stem growth reached control levels after up to 4Â yr only. Modelling suggested that organ and carbon reserve turnover rates play a key role for a treeâs responsiveness to environmental changes. Needle turnover rate was found to be most important to accurately model stem growth dynamics. We conclude that leaf area and its adjustment time to new conditions is the main determinant for radial stem growth of pine trees as the transpiring area needs to be supported by a proportional amount of sapwood, despite the growth-inhibiting environmental conditions
Emulsion sheet doublets as interface trackers for the OPERA experiment
New methods for efficient and unambiguous interconnection between electronic
counters and target units based on nuclear photographic emulsion films have
been developed. The application to the OPERA experiment, that aims at detecting
oscillations between mu neutrino and tau neutrino in the CNGS neutrino beam, is
reported in this paper. In order to reduce background due to latent tracks
collected before installation in the detector, on-site large-scale treatments
of the emulsions ("refreshing") have been applied. Changeable Sheet (CSd)
packages, each made of a doublet of emulsion films, have been designed,
assembled and coupled to the OPERA target units ("ECC bricks"). A device has
been built to print X-ray spots for accurate interconnection both within the
CSd and between the CSd and the related ECC brick. Sample emulsion films have
been extensively scanned with state-of-the-art automated optical microscopes.
Efficient track-matching and powerful background rejection have been achieved
in tests with electronically tagged penetrating muons. Further improvement of
in-doublet film alignment was obtained by matching the pattern of low-energy
electron tracks. The commissioning of the overall OPERA alignment procedure is
in progress.Comment: 19 pages, 19 figure
Measurement of the atmospheric muon charge ratio with the OPERA detector
The OPERA detector at the Gran Sasso underground laboratory (LNGS) was used
to measure the atmospheric muon charge ratio in the TeV energy region. We
analyzed 403069 atmospheric muons corresponding to 113.4 days of livetime
during the 2008 CNGS run. We computed separately the muon charge ratio for
single and for multiple muon events in order to select different energy regions
of the primary cosmic ray spectrum and to test the charge ratio dependence on
the primary composition. The measured charge ratio values were corrected taking
into account the charge-misidentification errors. Data have also been grouped
in five bins of the "vertical surface energy". A fit to a simplified model of
muon production in the atmosphere allowed the determination of the pion and
kaon charge ratios weighted by the cosmic ray energy spectrum.Comment: 14 pages, 10 figure
The detection of neutrino interactions in the emulsion/lead target of the OPERA experiment
The OPERA neutrino detector in the underground Gran Sasso Laboratory (LNGS)
was designed to perform the first detection of neutrino oscillations in
appearance mode through the study of oscillations. The
apparatus consists of an emulsion/lead target complemented by electronic
detectors and it is placed in the high energy long-baseline CERN to LNGS beam
(CNGS) 730 km away from the neutrino source. Runs with CNGS neutrinos were
successfully carried out in 2007 and 2008 with the detector fully operational
with its related facilities for the emulsion handling and analysis. After a
brief description of the beam and of the experimental setup we report on the
collection, reconstruction and analysis procedures of first samples of neutrino
interaction events
First events from the CNGS neutrino beam detected in the OPERA experiment
The OPERA neutrino detector at the underground Gran Sasso Laboratory (LNGS)
was designed to perform the first detection of neutrino oscillations in
appearance mode, through the study of nu_mu to nu_tau oscillations. The
apparatus consists of a lead/emulsion-film target complemented by electronic
detectors. It is placed in the high-energy, long-baseline CERN to LNGS beam
(CNGS) 730 km away from the neutrino source. In August 2006 a first run with
CNGS neutrinos was successfully conducted. A first sample of neutrino events
was collected, statistically consistent with the integrated beam intensity.
After a brief description of the beam and of the various sub-detectors, we
report on the achievement of this milestone, presenting the first data and some
analysis results.Comment: Submitted to the New Journal of Physic
SUMO modification of cell surface Kv2.1 potassium channels regulates the activity of rat hippocampal neurons
Voltage-gated Kv2.1 potassium channels are important in the brain for determining activity-dependent excitability. Small ubiquitin-like modifier proteins (SUMOs) regulate function through reversible, enzyme-mediated conjugation to target lysine(s). Here, sumoylation of Kv2.1 in hippocampal neurons is shown to regulate firing by shifting the half-maximal activation voltage (V1/2) of channels up to 35 mV. Native SUMO and Kv2.1 are shown to interact within and outside channel clusters at the neuronal surface. Studies of single, heterologously expressed Kv2.1 channels show that only K470 is sumoylated. The channels have four subunits, but no more than two non-adjacent subunits carry SUMO concurrently. SUMO on one site shifts V1/2 by 15 mV, whereas sumoylation of two sites produces a full response. Thus, the SUMO pathway regulates neuronal excitability via Kv2.1 in a direct and graded manner
CDK19 is disrupted in a female patient with bilateral congenital retinal folds, microcephaly and mild mental retardation
Microcephaly, mental retardation and congenital retinal folds along with other systemic features have previously been reported as a separate clinical entity. The sporadic nature of the syndrome and lack of clear inheritance patterns pointed to a genetic heterogeneity. Here, we report a genetic analysis of a female patient with microcephaly, congenital bilateral falciform retinal folds, nystagmus, and mental retardation. Karyotyping revealed a de novo pericentric inversion in chromosome 6 with breakpoints in 6p12.1 and 6q21. Fluorescence in situ hybridization analysis narrowed down the region around the breakpoints, and the breakpoint at 6q21 was found to disrupt the CDK19 gene. CDK19 was found to be expressed in a diverse range of tissues including fetal eye and fetal brain. Quantitative PCR of the CDK19 transcript from EpsteinâBarr virus-transformed lymphoblastoid cell lines of the patient revealed ~50% reduction in the transcript (p = 0.02), suggesting haploinsufficiency of the gene. cdk8, the closest orthologue of human CDK19 in Drosophila has been shown to play a major role in eye development. Conditional knock-down of Drosophila cdk8 in multiple dendrite (md) neurons resulted in 35% reduced dendritic branching and altered morphology of the dendritic arbour, which appeared to be due in part to a loss of small higher order branches. In addition, Cdk8 mutant md neurons showed diminished dendritic fields revealing an important role of the CDK19 orthologue in the developing nervous system of Drosophila. This is the first time the CDK19 gene, a component of the mediator co-activator complex, has been linked to a human disease
Chronic non-transmural infarction has a delayed recovery of function following revascularization
<p>Abstract</p> <p>Background</p> <p>The time course of regional functional recovery following revascularization with regards to the presence or absence of infarction is poorly known. We studied the effect of the presence of chronic non-transmural infarction on the time course of recovery of myocardial perfusion and function after elective revascularization.</p> <p>Methods</p> <p>Eighteen patients (mean age 69, range 52-84, 17 men) prospectively underwent cine magnetic resonance imaging (MRI), delayed contrast enhanced MRI and rest/stress 99m-Tc-tetrofosmin single photon emission computed tomography (SPECT) before, one and six months after elective coronary artery bypass grafting (CABG) or percutaneous coronary intervention (PCI).</p> <p>Results</p> <p>Dysfunctional myocardial segments (n = 337/864, 39%) were classified according to the presence (n = 164) or absence (n = 173) of infarction. Infarct transmurality in dysfunctional segments was largely non-transmural (transmurality = 31 ± 22%). Quantitative stress perfusion and wall thickening increased at one month in dysfunctional segments without infarction (p < 0.001), with no further improvement at six months. Despite improvements in stress perfusion at one month (p < 0.001), non-transmural infarction displayed a slower and lesser improvement in wall thickening at one (p < 0.05) and six months (p < 0.001).</p> <p>Conclusions</p> <p>Dysfunctional segments without infarction represent repetitively stunned or hibernating myocardium, and these segments improved both perfusion and function within one month after revascularization with no improvement thereafter. Although dysfunctional segments with non-transmural infarction improved in perfusion at one month, functional recovery was mostly seen between one and six months, possibly reflecting a more severe ischemic burden. These findings may be of value in the clinical assessment of regional functional recovery in the time period after revascularization.</p
Mediator and cohesin connect gene expression and chromatin architecture
Transcription factors control cell-specific gene expression programs through interactions with diverse coactivators and the transcription apparatus. Gene activation may involve DNA loop formation between enhancer-bound transcription factors and the transcription apparatus at the core promoter, but this process is not well understood. Here we report that mediator and cohesin physically and functionally connect the enhancers and core promoters of active genes in murine embryonic stem cells. Mediator, a transcriptional coactivator, forms a complex with cohesin, which can form rings that connect two DNA segments. The cohesin-loading factor Nipbl is associated with mediatorâcohesin complexes, providing a means to load cohesin at promoters. DNA looping is observed between the enhancers and promoters occupied by mediator and cohesin. Mediator and cohesin co-occupy different promoters in different cells, thus generating cell-type-specific DNA loops linked to the gene expression program of each cell.National Institutes of Health (U.S.) (Fellowship)Canadian Institutes of Health Research (Research Fellowship)National Institutes of Health (U.S.) (Grant R01 HG002668
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