bluuebunny/crossref_metadata_2025_split huggingface dataset:
TGFbeta signalling through Smad‐independent mechanisms in human lens cells
Authors: WORMSTONE IM , ELDRED JA , DAWES LJ , REDDAN JR , WANG L 
Published: September 2008

Abstract Purpose Previous work has shown that TGFß can induce matrix contraction and is likely to be regulated by Smad‐independent pathways. The present study investigated the role of the classical MAPK signalling and Rho kinase signalling pathways in TGFß mediated matrix contraction. Methods Cells from the human lens line FHL 124 were employed. Contraction was assessed using a patch assay, whereby the area covered by cells, was measured using imaging techniques. ERK activation was evaluated using western blot methods. To study SMAD2/3 distribution immunocytochemistry was used. Following a 24hr period of serum starvation, cells were maintained in the following conditions: Control medium +/– 10µM MEK inhibitor (U0126); 10ng/ml TGFß2 +/– U0126; Control medium +/– 10µM Rho kinase Inhibitor (Y–27632); 10ng/ml TGFß2 +/– Y–27632. The experimental duration for Smad2/3 staining was 2 hrs; western blots, 30mins & 24hr; patch contraction assays, 2‐3 days. Results Addition of 10ng/ml TGFß2 lead to a significant elevation of P‐ERK levels following both 30 min and 24hr periods of exposure. Addition of 10ng/ml TGFß2 to patch assays caused a significant contractile event to take place. Maintenance of cultures in the presence of 10µM U0126 or Y–27632 had no significant effect on patch area when added to control medium; however, when added in the presence of TGFß2, contraction was inhibited. With respect to cell population all groups were similar. Immunocytochemistry for SMAD2/3 showed clear nuclear translocation in response to 10ng/ml TGFß2; this signalling event was unaffected by U0126 and Y–27632. Conclusion MAPK and Rho kinase signalling play important roles in TGFß mediated matrix contraction of human lens cells.
Commercial interest
A novel cytocompatible thermoresponsive co‐polymer for corneal tissue engineering
Authors: HOPKINSON A , VARGHESE VM , KUMARY TV , DUA HS 
Published: September 2008

Abstract Purpose Using thermoresponsive polymers as a substrate to expand healthy sheets of transplantable cells for the immediate cover of wounds is an attractive novel treatment for ocular surface reconstruction. However, the cytocompatibility of thermoresponsive polymer and the effect on cell expansion must be assessed. Changes in the expression of secretory proteins in response to different microenvironments can be used to determine a more precise suitability of polymers in cell culture at molecular level. We assess the specific cytocompatibility of a novel thermoresponsive co‐polymer with the future goal of its use in corneal reconstruction. Methods Protein was extracted from conditioned medium of SV‐40 immortalised corneal cell line (CCL) cultured on both thermoresponsive polymer and tissue culture plastic. Proteins were separated by two‐dimensional gel electrophoresis and the resulting spot map profiles were differentially compared. Differences in the protein profiles were characterised by mass spectrometry and western blot analysis. Results Differential analysis of whole cell extract, and conditioned medium from CCL on plastic identified over 50 potentially secreted proteins. Mass spec characterisation confirmed these proteins as secretory, but also suggested healthy cell expansion was occurring. Comparison of CCL on plastic and thermoresponsive polymer revealed few differences suggesting the polymer did not effect cell expansion. Conclusion Thermoresponsive polymer is cytocompatible for CEC cell growth. This novel co‐polymer maybe a potential substrate for the future ex‐vivo engineering of transplantable cell sheets, for improved corneal reconstruction
Determining rates of visual field progression in glaucoma
Authors: CHAUHAN B 
Published: September 2008

Abstract Purpose To provide practical guidelines on detecting rates of visual field progression in glaucoma Methods Using a mixture of real patient data, computer simulation and statistical analysis, the frequency of visual field examinations for detecting various rates of visual field change were determined. Results Our results show that the ability to detect rates of visual field change depends critically on the magnitude of the change we wish to detect and the variability of visual fields. They also show that performing only one visual field per year will lead to failure to detect very significant visual field loss. The statistical power to detect clinically meaningful rates increases with 2 or 3 examinations per year Conclusion This study provides guidance to general ophthalmologists and glaucoma specialists on detecting rates of visual field progression in glaucoma.
Commercial interest
Myofibroblast and smooth muscle actin in scarred corneal stroma
Authors: AKHTAR S , ALMUBRAD TM , OGBUEHI KC , BRON AJ 
Published: September 2008

Abstract Purpose Mohan et al, (2003) hypothesised that after corneal injury, the site of injury is invaded by myofibroblasts (MFBCs) derived from dividing keratocytes. We report here the distribution of myofibroblasts (MFBCs), and cellular location of α‐‐ smooth muscle actin (α‐‐SMA) in scarred cornea. Methods Three penetrating keratoplasty buttons, two with post‐zoster scarring and one with hypertrophic dendriform epitheliopathy, were fixed in: 1) 2.5% glutaraldehyde containing cuprolinic blue for ultrastructural studies and 2) 4% paraformaldehyde for immunogold localisation. Primary α‐‐SMA and vimentin antibodies were localised by gold conjugates. Results Large numbers of MFBCs were observed in the sub‐epithelial collagenous pannus, posterior stroma and posterior collagenous layer in these corneas. The MFBCs were very large in size, lay in layers and occasionally fused with each other. Labelling for α‐SMA was very strong in all MFBCs. Labelling of α‐SMA was also observed in basal epithelial cells. MFBCs contained very prominent rough endoplasmic reticulum, vimentin filaments and large nuclei. Significantly, large proteoglycans (nm) and collagen fibrils (nm) were present around these cells. Conclusion It has been suggested that cytokynes released from the overlying epithelium act to maintain the myofibroblast phenotype (Jester et al 1999a, 1999b). We hypothesise that MFBCs synthesise large amounts of proteoglycan which disrupt the organisation of collagen fibrils and lead to the opacities of scarred cornea. The finding of labelling for α‐SMA in basal epithelial cells suggests that some epithelial cells may transform into MFBCs.
Geometric characterization of anterior segment in the hen’s eye
Authors: ALIO SANZ JL , PINERO LLORENS DP , SIREROL B , BATAILLE L 
Published: September 2008

Abstract Purpose To characterize the corneal geometry and the anatomical configuration of the anterior segment in the hen’s eye Methods 6 eyes of 3 brown leghorn hens with a mean age of 6 months were analyzed. The animals were anesthesized prior to the beginning of the measurements. In all cases, corneal geometry and anatomical configuration of the anterior segment were evaluated by means of the CSO and Visante OCT systems, respectively Results Mean corneal curvature in the 3‐mm central area was 69.35 ± 1.77 D, with a marked corneal astigmatism of a mean value of 3.00 ± 1.86 D. Mean corneal asphericity was ‐0.23 ± 0.41. Regarding central pachymetry, a mean value of 306.67 ± 46.76 microns was found. Anterior chamber had a mean diameter of 8.14 ± 0.13 mm and a mean associated sagittal depth of 2.33 ± 0.23 mm Conclusion The hen’s eye has a thin cornea, slightly prolate and with significant steepness, and an anterior chamber with a reduced diameter
Two distinct populations of corneal epithelial cells with limbal stem cell characteristics in the mouse
Authors: HOLAN V , KRULOVA M , POKORNA K , LENCOVA A , ZAJICOVA A , FILIPEC M 
Published: September 2008

Abstract Purpose Purpose. To detect and isolate cells with stem cell (SC) characteristics in the limbus of the mouse. Methods Methods. Limbal tissues from BALB/c mice were trypsin‐dissociated and separated on a Percoll gradient. Several fractions were isolated and characterized by Real‐time PCR for the presence of limbal SC markers and differentiation markers of corneal epithelial cells, by flow cytometry for the determination of the side‐population (SP) phenotype and for growth properties in vitro. Results Results. Cells retained in the 40% Percoll fraction and in the 80% Percoll fraction of the gradient were both enriched for populations with a high expression of SC markers ABCG2 and Lgr5 and also expressing the SP phenotype. However, the lightest fraction contained cells with the strongest spontaneous proliferative capacity and expressed the corneal epithelial differentiation marker K12. In contrast the densest fraction was K12 negative and contained small non‐spontaneously proliferating cells, which instead were positive for p63. Conclusion Conclusion. These findings demonstrate the presence of two distinct populations of corneal epithelial cells with limbal SC characteristics, based on differential expression of the keratin specific marker K12 and transcription factor p63, and suggest a difference in developmental stage of the two populations, with the K12‐ p63+ population being closer to the primitive limbal SC.
Depth profile study of molecular collagen structure in normal human cornea
Authors: KAMMA‐LORGER CS , BOOTE C , YOUNG RD , HAYES S , QUANTOCK AJ , MEEK KM 
Published: September 2008

Abstract Purpose To investigate the structure and organisation of corneal collagen as a function of stromal depth. Methods A strip extending from limbus to limbus along the superior to posterior meridian was cut from a normal human cornea and 100µm serial sections, parallel to the surface plane of the tissue, were obtained. Wide angle x‐ray diffraction experiments were performed in ID‐13 at the European Synchrotron Radiation Facility (ESRF) using a 25µm square beam. Each section was scanned at 450µm or 500µm steps. Additionally, another thin vertical strip (1mm thickness) was mounted in such a way that the beam was scanning edge‐on. Two scans were performed (1mm apart, each covering the depth of the cornea) from the anterior to the posterior surface at 25µm intervals. Data analysis provided information on the direction and degree of alignment of preferentially oriented collagen fibrils, as well as collagen intermolecular spacing (IMS). Results IMS is relatively stable throughout the depth of the central cornea, but is decreased in the posterior peripheral cornea. The degree of preferential alignment of fibrillar collagen is lower in the peripheral anterior part. In addition, the previously well‐documented tangential or annular collagen structure at the limbus is mainly restricted to the most posterior 100µm of stroma. At more superficial layers collagen preferred orientation tends to obtain an “x” shaped pattern, possibly forming part of the "anchoring fibril" structure alluded to previously in full‐thickness averaged x‐ray studies. Conclusion The preferred directions of the lamellae and IMS are depth‐dependent. The annulus at the limbus, which is thought to be implicated in the maintenance of corneal curvature, resides in the posterior limbus.
Leber´s stellate neuroretinitis. A case report
Authors: IDOIPE CORTA M , GIL ARRIBAS L , GARCIA‐MARTIN E , FERRER NOVELLA E 
Published: September 2008

Abstract Purpose A 49 years old man was admitted in our hospital with a 4‐day history on sudden loss of vision in the right eye. Methods Completed ocular examination, including visual acuity, anterior ocular segment, pupillary light reflex, intaocular pressure, ophthalmoscopic examination, visual fields, fluorescein angiography, system examination, including computed tomography and serologic test were done. Results We present a 49 years old man with sudden loss of vision who presented unilateral disc swelling. Five days later, the fundus examination reveled a macular star, and peripapillary retinal detachment. The fundus findings were normalized in a few weeks, leaving behind only retinal pigment epithelial defects and an inferior visual field defect. Infectious aetiology and others causes were rejected. Conclusion Leber´s idiopathic stellate neuroretinitis is un uncommon syndrome characterized by unilateral optic disc swelling followed by a macular star. It has a spontaneous resolution, and the aetiology is unknown.
Unilateral optic neuritis as presenting feature of acute hemorrhagic leukoencephalitis
Authors: SKET KONTESTABILE A , SEGA S , SVIGELJ V , HAWLINA M 
Published: September 2008

Abstract Purpose We report a case of 23 year‐old previously healthy female with acute hemorrhagic leukoencephalitis AHLE (Hürst disease) leading to lethal outcome within two weeks after presentation with unilateral visual loss and moderate upper respiratory tract infection with leukocytosis. Methods At presentation, VA was 0.3 RE and 1.2 LE with centrocecal defect on right eye, positive RAPD and dyschromatopsia. Optic nerve on fundus examination was normal. Neurological examination revealed right sided hyperreflexia and diminished right plantar response. Initial MRI revealed few focal periventricular hyperintensive lesions consistent with demyelination. Results Treatment started with 1g metilprednisolon i.v. for 3 days with good initial effect. Third day of treatment, VA in right eye was normal (1.2), RAPD was negative and dyschromatopsia improved. Three days later, altered consciousness with seizures developed, subsequently followed by right hemiparesis and coma. Cerebrospinal fluid pleocytosis was present. Follow‐ up MRI, obtained 4 days after initial one, revealed large areas of hemorrhagic necrosis in the left hemisphere.Despite continued high dose steroid treatment,antiviral prophylaxis and plasmapharesis, the disease progressed to death within days. Conclusion Acute hemorrhagic leukoencephalitis AHLE (Hürst disease) is a rare fulminant demyelinating disorder characterized by a fulminant clinical course due to hemorrhagic necrosis of the white matter associated with high rate of mortality. We present a case of this severe disease in which optic neuritis was presenting focal neurological deficit.
Acute painful ophthalmoplegia: infectious considerations
Authors: GOLNIK K 
Published: September 2008

AbstractAcute painful ophthalmoplegia may be caused by a number of processes including inflammation (giant cell arteritis, sarcoidosis, thyroid eye disease, idiopathic), vascular (aneurysm, hemorrhage, thrombosis, ischemia, fistula), tumor, and migraine. However, infectious causes (fungal, bacterial) of ophthalmoplegia may be life‐threatening if the diagnosis and treatment is delayed. Presenting symptoms and signs, differential diagnosis, and the appropriate treatment of these infectious entities will be discussed.
Microarray study of limbal epithelial crypt demonstrates its putative limbal stem cell niche characteristics
Authors: KULKARNI B , FOSTER T , YEUNG AM , TIGHE P , HOPKINSON A , DUA HS 
Published: September 2008

Abstract Purpose We have discovered a novel anatomical structure at the limbus, termed the Limbal Epithelial crypt (LEC). Previous anatomical and immunohistological studies suggest its potential as the limbal stem cell niche. We aim to characterise the differential gene expression of the LEC by microarrays with a view to finding putative stem cell marker(s). Methods Frozen tissue blocks of corneoscleral buttons dissected from cadaver eyes were cryosectioned. These tissue sections from different ocular surface regions were laser microdissected. Extracted RNA was amplified & hybridized to microarray chips. Raw data obtained with Genepix Pro6 software was filtered, normalized & analysed on BASE, TMeV & Jexpresspro software. Unpaired T‐Test, Significance Analysis of Microarrays and k‐means clustering were performed on the data. Quantitative gene expression analysis (qPCR) was performed on the genes of interest. Results 150 differentially expressed genes in the LEC were obtained(p ≤ 0.01). These genes were clustered according to stem cell related functions. qPCR was performed on the Integrin Beta 1 binding protein gene (ITGB1BP1). The protein expressed by this gene specifically binds to Beta 1 Integrin (ITGB1) which is a cell adhesion molecule, is predominantly an inhibitor of cell proliferation and has a role in maintaining stem cells in their niche. These genes were significantly expressed in the LEC (P=0.03). Conclusion The constituents of the LEC are stem cells with neighbouring niche cells, hence representing a Limbal Epithelial Stem Cell niche.
Distribution of BIG‐H3 protein in primary and recurrent granular corneal dystrophy
Authors: AKHTAR S , ALMUBRAD TM , OGBUEHI KC , BRON AJ 
Published: September 2008

Abstract Purpose Granular corneal dystrophy (GCD) is a hereditary disease involving the formation of opaque granular deposits in the corneal stroma. In the present project we investigated the distribution of big‐h3 protein in the cornea of primary GCD (PGCD), recurrent GCD (RGCD) and following epithelial debridement of RGCD. Methods The distribution of big‐h3 protein was studied in 2 PGCDs, 2 RGCDs, and in epithelial debridement specimens from 2 RCGD corneas. The corneas were fixed in 4% paraformaldehyde at 4°C and embedded in LR white under UV light. The primary antibody big‐h3 was located by secondary, goat anti‐rabbit antibody conjugated with gold. Results In normal cornea moderate distribution of big‐h3 was observed. In PGCD and RGCD corneas, strong labelling for big‐h3 was observed in deposits, within very thin microfilaments (nm) and at the inter‐lamellar junctions. Labelling was significantly higher in GCD Descemet’s membrane compared to normal Descemet’s membrane. Very strong labelling was observed in the deposits and in the cytoplasm of epithelial cells, of debridement specimens. Conclusion Big‐h3 protein, of epithelial origin, is thought to diffuse into the stroma to form stromal deposits. The presence of aggregates of the protein around keratocytes suggests that it is also produced by these cells. The presence of granular deposits and big‐h3 protein suggested that the protein migrated from limbal epithelium during resurfacing of host epithelium.
Anterior segment imaging with fourier‐domain optical coherence tomography system compared to time‐domain OCT
Authors: NOWINSKA A , WYLEGALA E , MILKA M , MANKOWSKI W 
Published: September 2008

Abstract Purpose To compare anterior segment measurements and morphology obtained from two anterior segment optical coherence tomography systems Methods 30 eyes of 30 healthy volunteers were recruited for the study. Central corneal thickness (CCT), trabecular‐iris angle (TIA) and angle opening distance (AOD 500) at the nasal and temporal angles were measured three times during one visit with two systems: Visante OCT (Carl Zeiss Meditec, Inc, Dublin, California, USA) and RTVue FD‐OCT Corneal Anterior Module system (Optovue, Inc. Fremont, CA, USC). Anterior segment morphology was assessed and compared. Results Mean values of CCT, TIA and AOD by Visante OCT and RTVue FD‐OCT were as follows: automatic CCT 535 ± 33,07 μm, 538 ± 31,82 μm; manual CCT 545 ± 30,91 μm, 542 ± 30,57 μm; nasal TIA 34,7± 9,5, 35,2 ± 8,9; temporal TIA 35,3 ± 8,5, 35,5 ± 9; nasal AOD 435 ± 95 μm, 444 ± 98 μm; temporal AOD 443 ± 103 μm, 452 ± 99 μm. There was no significant difference between mean values and they were highly correlated. Morphologic analysis of two OCT systems proved that Visante OCT has lower resolution, but in one anterior chamber image, all the anterior chamber structures are visible, while RTVue provides very precise information about small fragments of anterior chamber. Conclusion RTVue FD‐OCT provides accurate anterior segment parameters measurements and shows agreement wit OCT Visante.
Role of gap junction proteins in primary fiber cell elongation
Authors: GRAW J , PUK O , HORSCH M , BECKERS J 
Published: September 2008

Abstract Purpose During lens development, the elongation of primary fiber cells is the first crucial step after formation of the lens vesicle. According to characterized mouse mutants, this step depends on the activity of at least three genes: Maf, Pparbp and Gjf1, which we reported recently to encode a new connexin. In all these mutants, lens development is inhibited because the primary fiber cells do not elongate and the secondary fiber cells cannot form properly. The Gjf1 gene is altered in the mouse mutant Aey12 being characterized by microphthalmia, frequently associated with cataracts. Methods To identify downstream targets of Gjf1, we performed expression profiling in Gjf1Aey12 mouse embryos at 10.5 days of development. Results At this stage, the lens placode is beginning to form the lens vesicle and histological and morphological differences between wild‐type and mutant eyes are not yet obvious. We assumed that differences in gene expression might precede the morphological Aey12 phenotype. 31 genes were identified as up‐regulated. Among them, Fhod1 shows the highest level of induction; it encodes a Formin homology‐2 domain encoding protein. Another interesting up‐regulated gene is Rai16 (coding for the retinoic acid induced protein 16) providing a new link between lens development and retinoic acid signaling. Conclusion Since retinoic acid was shown recently to be necessary for lens differentiation in quails (Maden et al., 2007), the analysis of our Aey12 mutant might contribute to a better understanding of the role of retinoic acid and connexins during early lens development in mammals.
Cholesterol oxides, signal transduction and cell dysfunction in the lens: a bizarre love triangle
Authors: PEREIRA P , FERNANDES AF , FERREIRA J , BRITO A , GIRAO H 
Published: September 2008

Abstract Purpose Cholesterol oxides was implicated in the development of cataract. The purpose of this study is to elucidate the molecular mechanisms underlying cholesterol oxide‐induced cell dysfunction in the lens. Methods Lens epithelial cells (LEC) were incubated with 7‐ketocholesterol (7‐KC) and 25‐hydroxycholesterol (25‐OH). The effects of cholesterol oxides on cytoskeleton were evaluated by immunofluorescence microscopy. Activation of p38 MAPK and Akt was detected by Western Blot. LC3 processing was detected by Western Blot and immunofluorescence confocal microscopy and autophagy was assessed by the LC3 I/LC3 II ratio. Results Cells treated with 25‐OH show increased migration and formation of lamellipodia. These effects are reversed by inhibitors of PI3K and p38 MAPK. Consistently, 25‐OH induces activation of Akt and p38 MAPK in LEC. On the other hand, treatment of LEC with 7‐KC decreases Akt levels and up‐regulates autophagy. The increase in autophagy following treatment with 7‐KC is prevented by overexpressing a constitutive active Akt. Chemical inhibition of PI3K or overexpression of a dominant negative Akt also leads to stimulation of autophagy. Conclusion 7‐KC stimulates autophagy in LEC by promoting degradation of Akt and activation of mTOR. The two different cholesterol oxides (7‐KC, 25‐OH) have opposite effects in the regulation of Akt, resulting in increased autophagy (7‐KC) and increased cell migration (25‐OH). Accumulation of cholesterol oxides in lens may thus account for loss of cell transparency by interfering with cell migration and by deregulating autophagy in LEC. Supported by FCT grants POCI/SAU‐MMO/57216/2004 and POCI/SAU‐OBS/57772/2004.