Search results for " Confocal"
showing 10 items of 306 documents
The Stress-Inducible Protein DRR1 Exerts Distinct Effects on Actin Dynamics.
2018
Cytoskeletal dynamics are pivotal to memory, learning, and stress physiology, and thus psychiatric diseases. Downregulated in renal cell carcinoma 1 (DRR1) protein was characterized as the link between stress, actin dynamics, neuronal function, and cognition. To elucidate the underlying molecular mechanisms, we undertook a domain analysis of DRR1 and probed the effects on actin binding, polymerization, and bundling, as well as on actin-dependent cellular processes. Methods: DRR1 domains were cloned and expressed as recombinant proteins to perform in vitro analysis of actin dynamics (binding, bundling, polymerization, and nucleation). Cellular actin-dependent processes were analyzed in trans…
Chromatin organization regulates viral egress dynamics.
2017
Various types of DNA viruses are known to elicit the formation of a large nuclear viral replication compartment and marginalization of the cell chromatin. We used three-dimensional soft x-ray tomography, confocal and electron microscopy, combined with numerical modelling of capsid diffusion to analyse the molecular organization of chromatin in herpes simplex virus 1 infection and its effect on the transport of progeny viral capsids to the nuclear envelope. Our data showed that the formation of the viral replication compartment at late infection resulted in the enrichment of heterochromatin in the nuclear periphery accompanied by the compaction of chromatin. Random walk modelling of herpes s…
Herpes simplex virus 1 induces egress channels through marginalized host chromatin
2016
AbstractLytic infection with herpes simplex virus type 1 (HSV-1) induces profound modification of the cell nucleus including formation of a viral replication compartment and chromatin marginalization into the nuclear periphery. We used three-dimensional soft X-ray tomography, combined with cryogenic fluorescence, confocal and electron microscopy, to analyse the transformation of peripheral chromatin during HSV-1 infection. Our data showed an increased presence of low-density gaps in the marginalized chromatin at late infection. Advanced data analysis indicated the formation of virus-nucleocapsid-sized (or wider) channels extending through the compacted chromatin of the host. Importantly, co…
Modulation of mitochondriotropic properties of cyanine dyes by in organello copper-free click reaction
2017
Cyanine (Cy) dyes show a general propensity to localize in polarized mitochondria. This mitochondriotropism was used to perform a copper-free click reaction in the mitochondria of living cells. The in organello reaction of dyes Cy3 and Cy5 led to a product that was easily traceable by Forster resonance energy transfer (FRET). As determined by confocal laser scanning microscopy, the Cy3-Cy5 conjugate showed enhanced retention in mitochondria, relative to that of the starting compounds. This enhancement of a favorable property can be achieved by synthesis in organello, but not outside mitochondria.
The application of casein phosphopeptide and amorphous calcium phosphate with fluoride (CPP-ACPF) for restoring mineral loss after dental bleaching w…
2018
Abstract Objective To conduct an in vitro evaluation of the effect of casein-phosphopeptide and amorphous calcium phosphate with fluoride [CPP-ACPF] upon the calcium [Ca] and phosphorus [P] composition and morphology of dental enamel and dentin after the application of two bleaching agents: 37.5% hydrogen peroxide [HP]) and 35% carbamide peroxide [CP]. Materials and methods The crowns of 40 extracted human teeth were divided into four groups (n = 10 each). The crowns were sectioned along the cervical-incisal axis, and each half was embedded in acrylic resin, leaving a window 3 mm in diameter to explore the enamel or dentin (according to the study group involved). Groups 1a and 1b correspond…
Viral highway to nucleus exposed by image correlation analyses.
2018
AbstractParvoviral genome translocation from the plasma membrane into the nucleus is a coordinated multistep process mediated by capsid proteins. We used fast confocal microscopy line scan imaging combined with image correlation methods including auto-, pair- and cross-correlation, and number and brightness analysis, to study the parvovirus entry pathway at the single-particle level in living cells. Our results show that the endosome-associated movement of virus particles fluctuates from fast to slow. Fast transit of single cytoplasmic capsids to the nuclear envelope is followed by slow movement of capsids and fast diffusion of capsid fragments in the nucleoplasm. The unique combination of …
Detecting Protein Aggregation on Cells Surface: Concanavalin A Oligomers Formation
2009
A number of neurodegenerative diseases involve protein aggregation and amyloid formation. Recently evidence has emerged indicating small-transient prefibrillar oligomers as the primary pathogenic agents. Noteworthy, strict analogies exist between the behaviour of cells in culture treated with misfolded non-pathogenic proteins and in pathologic conditions, this instance together with the observation that the oligomers and fibrils are characterised by common structural features suggest that common mechanisms for cytotoxicity could exists and have to be perused in common interactions involved in aggregation.We here report an experimental study on ConcanavalinA (ConA) aggregation and its effect…
The learning curve, accuracy, and interobserver agreement of endoscope-based confocal laser endomicroscopy for the differentiation of colorectal lesi…
2012
Background: The endoscope-based confocal laser endomicroscopy (eCLE) system allows in vivo imaging of colorectal epithelium. Little is known about the learning curve for accurate interpretation of confocal images acquired with eCLE. Objective: To determine the learning curve of eCLE, its diagnostic accuracy, and the intra- and interobserver agreement for the differentiation of colorectal lesions. Design: Post hoc assessment of selected eCLE images. Setting: Academic centers. Patients: This study involved colonoscopic images from 47 patients. Main Outcome Measurements: Learning curve of eCLE, accuracy, and intraobserver and interobserver agreement. Methods: Three endoscopists received a shor…
In vivo confocal laser endomicroscopy of the human liver: a novel method for assessing liver microarchitecture in real time
2008
Background and study aims Confocal endomicroscopy is a unique novel tool for in vivo histology in humans. Due to limitations imposed by the form of the equipment and by sterilization workflows, its use has been limited to the gastrointestinal tract so far. We have developed a rigid miniaturized probe for confocal endomicroscopy of the human liver during laparoscopy. Patients and methods To assess the feasibility and potential clinical value of this new system (diameter 6.3 mm), 25 patients with liver disease were examined during routine minilaparoscopy under conscious sedation. Results Subsurface serial images (from surface to 250 microm) were generated in real time after fluorescein inject…
Corneal confocal scanning laser microscopy in patients with dry eye disease treated with topical cyclosporine
2017
Purpose To investigate the effect of cyclosporine on corneal ultrastructure and on major signs and symptoms of patients with dry eye disease.Patients and methods In this prospective cohort study, patients with dry eye disease were treated with a drop of cyclosporine 0.05% twice daily. Clinical evaluation was carried out at baseline and at months 1, 3, and 6. All patients completed the Ocular Surface Disease Index (OSDI) questionnaire, and tear film break-up time (BUT), fluorescein and lissamine green staining, and Schirmer test were carried out. In vivo confocal microscopy was also performed and epithelial cellular density, keratocyte activation, and subbasal plexus morphology were assessed…