bluuebunny/crossref_metadata_2025_split huggingface dataset:
Improvement of delivery of molecules into corneal endothelium using nanoparticles activated by femtosecond laser pulses
Authors: JUMELLE C , CAMPOLMI N , BERNARD A , PISELLI S , BAUBEAU E , MAUCLAIR C , GRANIER J , EGAUD G , GAIN P , THURET G 
Published: November 2013

AbstractPurpose NanoFemtoTransfection (NFT) is an innovative and promising technique of gene transfer (Nature nanotechnology 2011). It consists in permeabilizing cell membrane with nanoparticles (NP) of black carbon activated by Ti‐Sapphire femtosecond laser (FsL) in order to safely introduce molecule of interest. Initially described for cell suspension, we adapted during last year NFT to human corneal endothelial cells (EC) Aim: to further optimize NFT of EC in vitroMethods The model of NFT of calcein (a small molecule) in an EC line (B4G12, DMSZ, Germany) was used to optimize exposition times, fluence and Ti‐Saphir FsL beam movements in order to determine the best efficiency/toxicity ratio. Calcein transfer was observed with fluorescent microscopy (IX81, Olympus) and quantified by flow cytometry (FACSCalibur, BD, CA). Viability of EC was assessed by propidium iodide staining. Staining of actine with phalloidin and immunostaining of ZO‐1 was performed to study cell alteration caused by NFTResults With a static laser beam increasing FsL fluence and exposure time increased transfection efficiency but also EC mortality. The use of mobile FsL beam (with a XY micrometric stage, cheaper than a scanner), permitted an irradiation of the entire area of EC culture resulting in a noticeably increased efficiency that reached 38% of transfected EC and a limited toxicity (3%). Calcein positive EC presented the same phalloidin and ZO‐1 patterns as control ECConclusion Optimized NFT parameters are currently being tested in our laboratory to transfect plasmids. Grants: Fondation pour la Recherche Médicale, Fondation des Aveugles de France
Femtosecond laser cutting of multiple ultrathin corneal stromal lamellae for endothelial graft bioengineering
Authors: BERNARD A , HE Z , CAMPOLMI N , PISELLI S , FOREST F , PEOC'H MC , DUMOLLARD JM , THURET G , GAIN P 
Published: November 2013

AbstractPurpose The bioengineering of endothelial grafts (EG) is developing in several countries as an answer to the shortage of donor corneas worldwide. They are composed of a thin transparent biocompatible carrier evenly covered with tightly packed fully functional endothelial cells. Multiple options exist for the carriers. In our laboratory, we are focused on the optimization of human donor tissue because the well‐organised cornea donation in France is not the most limiting point. Aim: to present the preparation of ultrathin stromal lamellae from a single human corneaMethods Human organ‐cultured corneas unsuitable for graft were used either with our without deswelling in Dextran containing medium. They were mounted on a artificial anterior chamber and cut with a femtosecond laser (femto LDV Z6, Ziemer) with aplanation. Six horizontal parallel cuts were realized on a 9 mm diameter area without vertical section. Cornea was then trephine at 8.00mm on a punch trephine. Thickness map was obtained before and after the cut with an AS‐OCT equipped with a custom adapter for ex vivo corneas. Lamellae were separated under an operating video‐microscope and again observed with OCT. Quality of surface were analysed with scanning electron microscopy and multiphotonic confocal microscopy with SHG for collagen fiber imagingResults Seven regular lamellae of approximately 100μm were obtained on a single cornea and easily detached. Characterisation of surface quality is ongoingConclusion Femtosecond laser cutting of multiple lamellae on a single human cornea could allow obtaining a sufficient number of thin transparent and fully biocompatible carriers for EG bioengineering
Ferrara ring’s effects on corneal asphericity in keratoconus treatment
Authors: BONNEL S , LUSSATO M , FENOLLAND JR , SAIB N , RAMBAUD C , ABRIEU M , FROUSSART F , RIGAL‐SASTOURNE JC 
Published: November 2013

AbstractPurpose In keratoconus treatment, Ferrera® rings nomogram integrates the Q factor defining corneal asphericity. Our study deals with the effect of the implantation of Ferrerra rings on the Q factorMethods This retrospective and monocentric study involves forty eyes of thirty‐one patients suffering from keratoconus implanted with Ferrara® rings, with an uncorrected visual activity (UCVA), a best spectacle‐corrected visual acuity (BSCVA) and a videokerato‐topography, before and six months after surgery. The items followed are the UCVA, the BSCA, the average (Kmean) and maximum keratometry (Kmax) and the Q factor in the central 20° (Q20) and 40° (Q40). The analyse of the visual acuity (more or less than a line) enables to differentiate two groups of “good” and “bad” responders. The statistical tests are a Wilcoxon signed rank for pre and post operative comparisons, a Wilcoxon‐Mann Whithey for group comparisons, and a Spearsman for correlations.Results The average age of patients is 32,8 years with 83,1% of male gender. Twelve eyes were implanted with two rings and twenty‐eight with one. We find an average gain of 3.6 lines for UCVA, 1.1 lines for BSCVA and a decrease in Kmean of 1,4 dioptry. Q20 and Q40 are meaningly improved after surgery, but there is no significant difference between the studied groups. Lastly, before surgery, BSCVA is correlated to Q20, and Kmax to Q40. After surgery, UCVA is correlated to Q20 and Kmax to Q20. But the Kmax variation and the UCVA or BSCVA gains are not correlated.Conclusion Including the Q factor in the nomogram establishing ring’s choice improves the keratometric field but not the visual outcome.
Intracorneal foreign body in the confocal microscopy in vivo images
Authors: WOJCIK L , WYLEGALA E , SMEDOWSKI A 
Published: November 2013

AbstractPurpose Presentation of a patient with intreacorneal foreign body, differentiation in images of corneal confocal microscopy in vivo.Methods The patient admitted because of suspicion of post‐traumatic epithelial ingrown syndrome, to diagnosis and treatment, the following tests were performed: basic slit lamp examination, corneal scans using Scheimpflug camera (Pentacam, Oculus) and corneal confocal microscopy in vivo (Rostock Cornea Module, Heidelberg Engineering Retina tomograph III).Results On a slit lamp examination we described macroscopic limited opacity without evidence of inflammation. In the images from the Scheimpflug camera ‐ hiperreflective deposits obscuring other structures of the cornea. The corneal confocal microscopy images describes post‐traumatic corneal scar changes with clearly marked wound after the foreign body intake and single round conjunctival epithelial cells within the corneal stroma. In addition activation of keratocytes was observed. On about 350 microns depth, small deposits of number from few to several, with increased linear streak hyperreflexy and large, organized foreign body (piece of glass) of 1.5x0.7 mm diameter, irregular margins, with a similar hiperreflexy.Conclusion Corneal confocal microscopy is a useful tool for the detection and differentiation of corneal opacities of uncertain etiology.
Can we consider syphilitic uveitis as neurosyphilis? A retrospective analysis of lumbar puncture results in a cohort of syphilitic uveitis patients
Authors: LHOIR S , WILLERMAIN F , JANSEN J , LIBOIS A , VAN CALSTER J , CASPERS L , MAKHOUL D 
Published: November 2013

AbstractPurpose To investigate the cerebral spinal fluid (CSF) composition of patients with syphilitic uveitis (SU) and classify SU as neurosyphilis or not.Methods Retrospective study of SU patients from 2 Belgian university centers from 2003 to 2012. The diagnosis of SU was based on a positive syphilitic serology (FTA‐Abs and RPR) and a treatment response. Charts were reviewed for uveitis anatomical location, HIV status, serum RPR and CSF findings. Diagnosis criteria for neurosyphilis were: positive CSF‐RPR and/or CSF‐white blood cells (WBC) ≥ 5/μl for HIV‐, and positive CSF‐RPR and/or CSF‐WBC ≥ 20/μl for HIV+.Results 20 patients were included. 5 were HIV+. There were 3 papillitis, 10 posterior uveitis and 7 panuveitis. Cells were found in CSF in all patients (mean 40/μl) and were predominantly lymphocytes. Mean CSF‐proteins, lactate and glucose were 0.57g/l, 17mg/dl and 62mg/dl respectively. No differences were found between HIV+ and – patients. CSF‐RPR was + in 2/5 (40%) of HIV+ and in 11/15 (73%) of HIV‐ with no statistical difference.Conclusion In our series, all patients had a CSF cellular reaction, mostly lymphocytic. This strongly suggests that the blood brain barrier is disrupted during syphilitic uveitis. However, a positive RPR was not observed in all CSF. Despite this discrepancy, following our criteria, 100% of our patients could be classified as neurosyphilis.
Adaptive optics imaging of retinal vasculitis
Authors: ERRERA MH , PAQUES M 
Published: November 2013

AbstractPurpose To document perivascular infiltrates in the retina using high resolution flood illumination adaptive optics (AO) imaging in patients with retinal vasculitis.Methods The charts of five subjects with retinal vasculitis showing evidence of perivascular infiltrates by means of AO imaging (rtx1 camera, Imagine Eye, Orsay, France) were reviewed.Results Diagnoses included primary idiopathic retinal vasculitis (n=1), Lyme’s disease (n=2), TB related retinal vasculitis (n=1) and idiopathic retinitis, vasculitis and aneurysm (IRVAN; n=1). Using AO imaging, perivascular infiltrates appeared as discrete, linear areas of opacification along the veins, often surrounding the venous narrowing. AO imaging was more sensitive that either fundus photographs or fluorescein angiography for their detection. Follow‐up examinations (n=3) showed a changing pattern of narrowing of the vessel lumen, and eventual disappearance of these perivascular infiltrates. In the IRVAN case, inflammatory infiltrates were found around arterioles and veins as well, suggesting an inflammation induced‐phenomenon.Conclusion AO imaging has a higher sensitivity than fundus photographs or fluorescein angiography to detect perivascular inflammatory infiltrates. Vascular inflammation is strongly linked to changes in vascular lumen diameter. AO imaging may have a particular interest when diagnosis and following subjects with retinal vasculitis.
Swept source‐optical coherence tomography analysis of choroidal involvement in patients with uveitis
Authors: DIWO E , RAZAVI S , LEHOANG P , BODAGHI B 
Published: November 2013

AbstractPurpose Swept source OCT (SS‐OCT) is currently the best visualization technique of the choroid. The aim of our study is to describe its thickness and vascular structures in patients with choroiditis.Methods We have completed a choroidal map with SS‐OCT, using 12‐mm horizontal vertical scans through the macula, for a group of patients with choroiditis, referred to Pitie Hospital, from October 2012 to March 2013.Results Eight patients have been included with a sex ratio of 1, an average age of 56.5 years : 5 birdshot retinochoroidopathies (BRC), 1 multifocal choroiditis , 1 Vogt‐Koyanagi‐Harada (VKH) disease, 1 acute posterior placoid chorioretinitis concomitant with a syphilitic seroconversion. In four active BRC, the average choroidal thickness was increased to 339 µm (389 µm ±41.93), among whom one BRC patient experienced a decrease of 30 µm in choroidal thickness immediately after corticotherapy. On the contrary the fifth patient, an advanced form of BRC, showed signs of atrophy with a choroidal thickness of 194 µm. Choroidal thickness in the VKH patient was unchanged in both eyes. There was a significant asymmetry in the patient with syphilitic chorioretinitis, with a choroidal thickness of 334 µm in the affected eye versus 387 µm in the contralateral eye. Yet, we observed that Haller’s layer of the affected eye was harder to visualize, due to possible inflammatory infiltrationsConclusion These choroidal maps are among the first choroiditis cases explored by SS‐OCT, first step of a mapping project which we wish to amplify in order to determine a possible threshold of choroidal thickness indicating a subclinical relapse. This could make SS‐OCT an additional tool besides ICG angiography for the exploration of the choroid.
Femtosecond laser capsulotomy versus manual technique: A clinico‐morphological study of the resected capsules
Authors: MEYER P , PRUENTE CH 
Published: November 2013

AbstractPurpose To compare resected lens capsules created by femtosecond laser system with the conventional manual techniques.Methods A prospective, observational case series of 6 patients who underwent capsulotomies performed by an optical coherence tomography‐guided femtosecond laser compared with 6 patients which capsulotomies created with the conventional manual technique. The resected capsules were evaluated histologically. Electron microscopy was performed to examine their capsule edge.Results Histologically the capsule edges produced by manual were smooth and continuous however performed by femtolaser the capsule edges showed small microgrooves in high magnification. Electronmiocroscopically these microgrooves presented as pointed serrated structure like a lacerated jeans.Conclusion The femtosecond laser produced capsulotomies were clinically more precise in diameter and in size however their capsules showed irregular stripped edges than those created with the conventional manual technique. The microgrooves were produced by the laser pulses. A certain weakness of the laser produced edges is to expect and needs more clinical experiences.
Metastases inhibition and cellular damage in melanoma cells irradiated with proton beam
Authors: ROMANOWSKA DIXON B , KEDRACKA‐KROK S , SKALNIAK L , JURA J , ZUBA‐SUMA E , JASINSKA K , PAWLAK A , SOWA U , OLKO P , URBANSKA K , ELAS M 
Published: November 2013

AbstractPurpose Proton beam therapy has been used in clinic with a great success, however, not many experimental studies has been performed. The goal of our study was to determine the cellular response to low, sublethal doses of proton beam irradiation, in particular DNA damage, cell cycle arrest, changes in expression of proteins, and effect on metastases in vivo.Methods BLM cells were irradiated with 1‐7 Gy of proton beam irradiation. The source of the 58 MeV proton beam was the AIC‐144 cyclotron at Institute of Nuclear Physics, Polish Academy of Sciences, Kraków. Tumors of Bomirski Hamster Melanoma (BHM) implanted into the anterior chamber of the hamster eye grew aggressively and completely filled the anterior chamber within 8‐10 days. Metastases, mainly in the lung, were found in 100% of untreated animals 30 days after enucleation. The protons were accelerated using AIC‐144 isochronous cyclotron, operating at 60 MeV and BHM tumors located in the anterior chamber of the eye were irradiated with 10 Gy, for the depth of 3.88 mm.Results Slow accumulation of damage was observed reflected in slowing of the proliferation rate, and increase in caspases activity with time. The number of cells in G2/M and >2n increased with proton beam dose. Proton beam irradiation caused upregulation of proteins involved in: DNA repair, RNA functioning (i.e. stress granule and P‐bodies components), apoptosis and survival processes and downregulation of enzymes engaged in glycolysis. Of particular interest was heavy downregulation of vimentin (2.4 times), involved in structural integrity of cells and tissues, adhesion and migration, and other processes. Irradiation led to changes in cell migratory properties. Proton beam irradiation caused inhibition of tumor growth by about 10 days and inhibition of metastastic spread.
Cases of choroidal metastasis revealing lung cancer
Authors: BARCATALI M , MIOCQUE S , HADDAD M , MOURIAUX F 
Published: November 2013

AbstractPurpose Choroidal metastases are the most common intraocular malignant tumor but rarely reveal cancer. However the role of the ophthalmologist is central to diagnose this disease. Ophthalmological examination, optical coherence tomography, ultrasound and angiography may help for differentiate metastasis to other choroidal tumors. The aim to this presentation is to describe to identical cases of choroidal metastasis revealing lung cancer.Methods We report 2 cases, a 53‐year‐old woman and a 63‐year‐old man, with no prior history of neoplasia who consulted for decreased visual acuity of the right eye. In both, eye fundus examination reveals a suspicious yellowish choroidal lesion. Fundus examination, optical coherence tomography (confirming the serous retinal detachment), ultrasound (confirming the plan or thick appearance) and angiography (showing blocking defect in early stage and hyperfluorescence with pin points in late stage for fluorescein angiography and diffuse hypofluorescence in late stage for indocyanin angiography) led us to the conclusion of choroidal metastasis. Then, next investigations were mammography (for woman) and body scan (for both). The results reveal an adenocarcinoma of the lung for both cases.Conclusion Choroidal metastases are rarely inaugural of a cancer but an early diagnosis is essential. The role of ophthalmologist is central to the diagnosis and both cases allow remembering the main diagnostic criteria.
PKC activation affects, via ELAVL1/HuR protein, VEGF expression in pericytic/endothelial coculture
Authors: AMADIO M , OSERA C , LUPO G , MOTTA C , DRAGO F , GOVONI S , PASCALE A 
Published: November 2013

AbstractPurpose To explore whether, following direct contact, there is a mutual influence between pericytes (PC) and endothelial cells (EC), and to establish whether PKC activation, a condition associated with hyperglycaemia in diabetic retinopathy (DR), can affect, via the mRNA‐binding ELAVL1/HuR protein, VEGF expression.Methods Retinal PC and EC were cultured separately (MONO) or in direct contact (1:1 ratio; coculture; COCU), and exposed or not to PKC activator phorbol esthers (100 nM PMA, 15 min.). PKCβI/βII, HuR, VEGF proteins were analysed by Western blotting; VEGF secretion in cell culture medium was evaluated by ELISA. VEGF mRNA analysis was performed by real‐time qPCR. Immunocytochemistry was performed to evaluate PMA‐induced HuR translocation.Results In MONO, VEGF mRNA/protein basal levels are higher in PC than EC. In COCU, VEGF protein amount is up‐regulated in EC and down‐regulated in PC, respectively. VEGF release is favoured by physical contact between PC and EC and further increased by PMA exposure. PKCβ activation induces HuR translocation from nucleus to cytoplasm, and it increases the protein levels of the kinase itself, HuR and VEGF in PC and EC in both culture conditions.Conclusion VEGF expression varies in function of the cell culture conditions. In all cultures, PKC activation increases PKCβI/βII, HuR and VEGF protein levels, changes which may occur also at very early stages of DR. The COCU model may be useful to study PC/EC interactions in both physiological and pathological conditions.
Macro and microglial retinal cells in rat organotypic cultures
Authors: GALLEGO B , ROJAS B , SALAZAR JJ , RAMíREZ AI , DIERSTEIN M , DE HOZ R , TRIVIñO A , RAMíREZ JM , UEFFING M , ARANGO‐GONZáLEZ B 
Published: November 2013

AbstractPurpose Organotypic retinal cultures constitute a useful tool to perform preclinical drug testing. The aim of the present study was characterize the morphologic changes in macro‐ and microglial cells in this culture system.Methods Retinas were isolated from 7‐day‐old Crl:CD(SD) rats with the retinal pigment epithelium attached. Retinal explants were cultured for 4, 10, and 14 days (DIV4, DIV10, and DIV14). Cryosections and whole‐mounts of age‐matched control and cultured retinas were used to analyze macro‐ and microglial cells by immunostaining using antibodies against GFAP, vimentin and CD11‐b.Results In comparison with in vivo, in DIV4 and DIV10 cultures, GFAP‐positive astrocytes were more robust, the astrocytic network being thicker in some retinal areas while in other regions astrocytes were sparsely distributed. Few thin GFAP‐positive astrocytes were observed at DIV14. Müller cells in the cultures exhibited GFAP up‐regulation in comparison with in vivo retinas. CD11‐b+ microglial cells at DIV4 and DIV10 showed more robust somas and thicker and more retracted processes than in vivo. Overall, at DIV14 CD11‐b+, microglial cells exhibited a rounded morphology.Conclusion In rat organotypic culture both macro‐ and microglial cells showed progressive changes: i) reactive macrogliosis, ii) rearranged astrocytic distribution, iii) GFAP up‐regulation in Müller cells, and iv) microglial activation. Given that this glial response is a hallmark of several retinal diseases, organotypic retinal culture is a valuable resource for future investigations in retinal degenerative processes and therapy.
A new automatic method for microglial‐cell quantification in whole‐mount mouse retinas
Authors: DE HOZ R , GALLEGO BI , ROJAS B , RAMIREZ AI , SALAZAR JJ , TRIVIÑO A , DE GRACIA PACHECO P , RAMIREZ JM 
Published: November 2013

AbstractPurpose To study the proliferative microglial behavior in a laser‐induced ocular hypertension (OHT) model with an automatic method that allows the assessment of the number of microglial cells and comparisons of the results with those found by direct human observation.Methods Albino Swiss mice were divided into: age‐matched control, n=6; and lasered, n= 6. Retinal whole‐mounts were immunostained with anti‐Iba1. In Matlab, new algorithms of segmentation and control of distances were developed to determine the number of Iba‐1+ cells. The automatic results were compared with those from direct human observation of the images.Results The algorithm method automatically detected the number of Iba‐1+ retinal cells in the inner and outer plexiform layers, both in naïve and OHT retinas. The number of cells present in the image samples was not an obstacle for the program to run properly. The time required for counting Iba 1+ cells decreased from the human guide to the program‐based counting method from days to one hour. the results showed a strong correlation between automatic and manual methods (Pearson correlation test, R = 0.979; P=0.000 and R= 0.942; P=0.000 for outer and inner plexiform layer, respectively) indicating the reliability of the automatic counting.Conclusion A new consistent and fast algorithm method was developed with Matlab to quantify Iba‐1+ microglial cells as well as cellular density maps through retinal whole‐mounts, in both naïve and OHT. Through this new automatic method, a larger set of images or samples could be included in future studies to analyze the behavior of microglial cells under proliferative conditions.