Search results for "focal"

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…

The still under-investigated role of cognitive deficits in PML diagnosis

2017

Background: Despite cognitive deficits frequently represent the first clinical manifestations of Progressive Multifocal Leukoencephalopathy (PML) in Natalizumab-treated MS patients, the importance of cognitive deficits in PML diagnosis is still under-investigated. The aim of the current study is to investigate the cognitive deficits at PML diagnosis in a group of Italian patients with PML. Methods: Thirty-four PML patients were included in the study. The demographic and clinical data, the lesion load and localization, and the longitudinal clinical course was compared between patients with (n = 13) and without (n = 15) cognitive deficit upon PML suspicion (the remaining six patients were asy…

E-beam crosslinked nanogels conjugated with monoclonal antibodies in targeting strategies

2017

Abstract Poly(N-vinyl pyrrolidone)-based-nanogels (NGs), produced by e-beam irradiation, are conjugated with monoclonal antibodies (mAb) for active targeting purposes. The uptake of immuno-functionalized nanogels is tested in an endothelial cell line, ECV304, using confocal and epifluorescence microscopy. Intracellular localization studies reveal a faster uptake of the immuno-nanogel conjugate with respect to the ‘bare’ nanogel. The specific internalization pathway of these immuno-nanogels is clarified by selective endocytosis inhibition experiments, flow cytometry and confocal microscopy. Active targeting ability is also verified by conjugating a monoclonal antibody which recognizes the αv…

2018

Abstract Purpose To report on a case of recurrence of paraproteinemic keratopathy (PPK) associated with monoclonal gammopathy after bilateral penetrating keratoplasty. Observations Penetrating keratoplasty was performed on both eyes of a 45-year-old man due to bilateral progressive corneal stromal clouding. Recurrence of the corneal stromal opacities accompanied by a decrease in visual acuity was observed on slit-lamp examination already two years after penetrating keratoplasty. Confocal laser scanning microscopy (CLSM) of the corneal grafts performed three years after penetrating keratoplasty showed bilateral morphological changes identical to that found in the patient's corneas prior to p…

Confocal scanning laser ophthalmoscopy versus modified conventional fundus camera for fundus autofluorescence.

2016

Fundus autofluorescence (FAF) is a noninvasive imaging method to detect fundus endogenous fluorophores, mainly lipofuscin located in the retinal pigment epithelium (RPE). The FAF provides information about lipofuscin distribution and RPE health, and consequently an increased accumulation of lipofuscin has been correlated with ageing and development of certain retinal conditions. Areas covered: An exhaustive literature search in MEDLINE (via OVID) and PUBMED for articles related to ocular FAF in retinal diseases and different devices used for acquiring FAF imaging was conducted. Expert commentary: This review aims to show an overview about autofluorescence in the RPE and the main devices use…

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…

Could the Calcium Silicate-Based Sealer Presentation Form Influence Dentinal Sealing? An In Vitro Confocal Laser Study on Tubular Penetration

2021

Dentinal tubule penetration influences root canal treatment sealing. The aim of this study was to compare dentinal penetration of two clinical presentations of silicate-based sealers using confocal laser. Sixty single-rooted human teeth from 50–70 year-old patients extracted for orthodontic/periodontal reasons were used. Canals were prepared using Mtwo system up to 35/0.04, with 5.25% NaOCl irrigation and final irrigation using 17% EDTA. Teeth were randomly assigned into study groups (ni = 20): EndoSequence BC sealer (ES, group 1), BioRoot RCS (BR, group 2); and a control group (nc = 20) with AH Plus (AHP). Root canals were obturated with 35/0.04 gutta-percha (single-cone technique). The sa…