The prevalence of activated protein C (APC) resistance and factor V Leiden is significantly higher in patients with retinal vein occlusion without general risk factors. Case-control study and meta-analysis.

Several small case-control studies have investigated whether factor V Leiden (FVL) is a risk factor for retinal vein occlusion (RVO) and generated conflicting data. To clarify this question we performed a large two-centre case-control study and a meta-analysis of published studies. Two hundred seven consecutive patients with RVO and a control group of 150 subjects were screened between 1996 and 2006. A systematic meta-analysis was done combining our study with further 17 published European case-control studies. APC resistance was detected in 16 out of 207 (7.7%) patients and eight out of 150 (5.3%) controls. The odds ratio (OR) estimated was 1.49 with a (non-significant) 95% confidence interval (CI) of 0.62-3.57. The meta-analysis including 18 studies with a total of 1,748 patients and 2,716 controls showed a significantly higher prevalence of FVL in patients with RVO compared to healthy controls (combined OR 1.66; 95% CI 1.19-2.32). All single studies combined in the meta-analysis were too small to reliably detect the effect individually. This explains the seemingly contradictory data in the literature. In conclusion, the prevalence of APC resistance (and FVL) is increased in patients with RVO compared to controls, but the effect is only moderate. Therefore, there is no indication for general screening of factor V mutation in all patients with RVO. We recommend this test to be performed in patients older than 50 years with an additional history of thromboembolic event and in younger patients without general risk factors like hypertension.

Glial cell-mediated spread of retinal degeneration during detachment: a hypothesis based upon studies in rabbits.

In human subjects with peripheral retinal detachments, visual deficits are not restricted to the detached retina but are also present in the non-detached tissue. Based upon studies on a rabbit model of rhegmatogenous retinal detachment, we propose a glial cell-mediated mechanism of spread of retinal degeneration into non-detached retinal areas which may also have importance for the understanding of alterations in the human retina. Both detached and attached portions of the rabbit retina display photoreceptor cell degeneration and cystic degeneration of the innermost layers. An inverse mode of photoreceptor cell degeneration in the attached tissue suggests a disturbed support of the photoreceptor cells by Müller cells which show various indications of gliosis (increased expression of intermediate filaments, cell hypertrophy, decreased plasma membrane K(+) conductance, increased Ca(2+) responsiveness to purinergic stimulation) in both detached and attached tissues. We propose that gliotic alterations of Müller cells contribute to the degeneration of the attached retina, via disturbance of glial homeostasis mechanisms. A down-regulation of the K(+) conductance of Müller cells may prevent effective retinal K(+) and water clearance, and may favor photoreceptor cell degeneration and edema development.

Upregulation of extracellular ATP-induced Müller cell responses in a dispase model of proliferative vitreoretinopathy.

PURPOSE: To test whether in an animal model of proliferative vitreoretinopathy (PVR) the Müller glial cells displayed an upregulation of purinergic P2 receptor-mediated responses. METHODS: PVR was induced by intravitreal injection of the proteolytic enzyme, dispase, in the eyes of adult rabbits. The developing PVR was examined ophthalmoscopically. After 3 weeks, small retinal pieces were wholemounted and used for calcium imaging, freshly dissociated Müller cells were subjected to calcium imaging, and patch-clamp recordings were made. The presence of P2 receptor-mediated Ca(2+) responses was determined both directly--that is, fluorometrically--and indirectly, by electrophysiological recording of Ca(2+)-activated K(+) currents. RESULTS: According to earlier observations in another model of retinal detachment and PVR, the reactive Müller cells displayed hypertrophy, downregulation of inwardly rectifying K(+) currents, and depolarization of the resting membrane potential, all dependent on the severity of the PVR. Further, significant PVR-induced increase was observed in the number of Müller cells responding to adenosine 5'-triphosphate (ATP), with a transient elevation of their [Ca(2+)](i). If isolated Müller cells were exposed to ATP, 13% of the control cells, but 29% (moderate PVR) or 53% (massive PVR) of the reactive cells, showed fluorometric Ca(2+) increases. An increase of Ca(2+)-activated K(+) currents was measured in 11% of the control cells, but in 83% (moderate PVR) and 90% (massive PVR) of the reactive cells. Confocal images of retinal wholemounts revealed similar results. Because similar responses were elicited by uridine triphosphate (UTP), the dominant involvement of metabotropic (P2Y type) purinergic receptors is suggested. CONCLUSIONS: An upregulation of purinergic receptors is part of the reactive changes of Müller cells during PVR. It is suggested that ATP-evoked Ca(2+) responses may support the proliferation of Müller cells during PVR.

Signaling pathways involved in PDGF-evoked cellular responses in human RPE cells.

We examined whether PDGF may directly stimulate the expression of VEGF by retinal pigment epithelial (RPE) cells in vitro, and the involvement of three signal transduction pathways in the regulation of PDGF-evoked cell proliferation, migration, and production of VEGF-A was investigated. PDGF stimulated the gene and protein expression of VEGF-A by RPE cells, and increased cell proliferation and chemotaxis. PDGF activated all signaling pathways investigated, as determined by increased phosphorylation levels of ERK1/2, p38, and Akt proteins. The three signaling pathways were involved in the mediation of PDGF-evoked cell proliferation, while p38 and PI3K mediated cell migration, and PI3K mediated secretion of VEGF-A. In addition to VEGF-A, the cells expressed mRNAs for various members of the VEGF family and for their receptors, including VEGF-B, -C, -D, flt-1, and KDR. The data indicate that PDGF selectively stimulates the expression of VEGF-A in RPE cells. PDGF evokes at least three signal transduction pathways which are differentially involved in various cellular responses.

Vitreoretinal endoscope for the assessment of the peripheral retina and the ciliary body after large retinectomies in severe anterior PVR.

PURPOSE: We evaluated endoscopically the changes of the peripheral retina and the ciliary body after large retinectomies. METHODS: The peripheral retina and the ciliary body of 5 patients with anterior proliferative vitreoretinopathy after large retinectomies (> 180 degrees) were visualized endoscopically. RESULTS: The endoscope allowed a complete assessment of the peripheral retina and the ciliary body. Even the 12 o'clock part of the eye could be evaluated with the 30 degrees optic. The peripheral retina, a fibrosis and a detachment of the ciliary body can be visualized and assessed. The cause of the postoperative hypotony after large retinectomies is mainly related to fibrosis and detachment of the ciliary body. Surgeons can expect a postoperative hypotony if fibrosis and a large detachment of the ciliary body is seen during surgery with the help of an endoscope. CONCLUSION: Endoscopic vitreoretinal visualization of the pars plana and ciliary body is a useful additional technique after large retinectomies in the treatment of anterior PVR.

Elevated proto-oncogene and collagen mRNA expression in PVR retinas.

PURPOSE: Retinal detachment is often accompanied by proliferation and migration of retinal cells and by increased synthesis of structural proteins, known as proliferative vitreoretinopathy (PVR). Herein we investigate the messenger RNA (mRNA) expression of proto-oncogenes responsible for cell proliferation and of structural proteins that have a role in membrane formation. METHODS: Retinal samples were obtained from patients undergoing vitreoretinal surgery for the treatment of retinal detachment complicated by PVR. Normal human control retinas were obtained from cornea donors. The mRNA expression of the proto-oncogenes c- myc, c- fos and the proliferation marker Ki67, as well as of collagen type III and type IV, were investigated using the ribonuclease protection assay. RESULTS: Ki67 mRNA expression was not detectable in either sample type, but c- fos and c- myc mRNA expression was found in normal and PVR retinas. Whereas the expression of c- myc showed a marginal increase, the up-regulation in c- fos expression was strongly significant (5.07-fold). The mRNA of collagen type III was detectable at widely varying levels in all the PVR retinas but was found in only 2 of the 16 analysed normal samples. Collagen type IV mRNA was expressed in both PVR and control samples but was higher (2.21-fold) in the PVR retinas. CONCLUSIONS: These results indicate that an up-regulation of the proto-oncogene c- fos occurs in human PVR retinas. An increase in mRNA expression of collagen types III and IV takes place simultaneously. These changes in mRNA expression appear to be mainly connected to the initiation of cell proliferation, dedifferentiation and formation of tractional membranes.

Determination of the solubility of perfluorocarbon liquids in silicone oil in vitro and in vivo.

PURPOSE: To investigate the solubility of perfluorocarbon liquids (PFCL) in silicone oil. METHODS: Forty-eight samples of silicone oil (1,300 mPas, n = 22; 5,000 mPas, n = 26) were analyzed for dissolved fluorocarbon molecules after surgical removal from patients who had initially undergone vitreoretinal surgery with (n = 41) and as control without (n = 7) the use of perfluorodecalin in headspace gas chromatography. In vitro, the solubility of three different PFCL-perfluorooctane (PFO), perfluorodecalin (PFD), and fluoromethylcyclohexane (FMCH)-in silicone oil of various viscosities was determined. The diffusion phenomena during a direct exchange were studied. RESULTS: In 39 of 41 silicone oil samples removed from patients who had undergone vitreoretinal surgery with the use of PFD, small amounts of dissolved perfluorocarbons could be detected. The mean value in 5,000-mPas silicone oil was 939.0 x 10-4 m/% and in 1,300-mPas silicone oil was 322.75 x10(-4) m/%. No perfluorocarbon molecules were found in seven control patients. In vitro, the following maximum solubilities in 1,000-mPas silicone oil were measured at room temperature: PFO, 3.2 m/%; PFD, 5.1 m/%; and FMCH, 10.3 m/%. The maximum values measured in 5,000-mPas silicone oil were PFO, 3.3 m/%; PFD, 5.7 m/%; and FMCH, 8.5 m/%; and in 100-mPas silicone oil were PFO, 2.4 m/%, and PFD, 5.1 m/%. CONCLUSION: Perfluorocarbon liquids dissolve in silicone oil. This may lead to transient formation of "heavy silicone oil," but no stable heavy silicone oil can be created adding PFCL. Intraocularly, retained PFCL vanish in silicone oil and are removed during silicone oil removal.

Recovery from hepatic retinopathy after liver transplantation.

BACKGROUND: In chronic liver disease the neuroglial cells may be affected by neurotoxic metabolites which, in turn, could be expected to affect neuronal functions. The aim of this study was to evaluate the electroretinograms (ERG) from patients before and after liver transplantation, in order to study possible functional changes of the retina. METHODS: Twelve patients with liver cirrhosis underwent routine ophthalmological examination and ERG before and after successful liver transplantation. Laboratory parameters, including ammonia, aspartate aminotransferase (AST), bilirubin, and cholinesterase, were compared. Patients were grouped according to the Child classification: three patients were Child A, six were Child B, three were Child C. RESULTS: Most obvious ERG abnormalities were found in patients with cirrhosis Child C. Before transplantation 7 of 21 ERG parameters were out of the normal range, but in the follow-up examination after transplantation only one parameter was not within the normal range. Significant ( P<0.05) postoperative improvements were found for the latencies of scotopic, mesopic and photopic b-waves and mesopic a-waves and for the photopic implicit time. Patients in the Child B group revealed less changes in the ERG. Before the transplantation only one parameter of the ERG was out of the normal range. All postoperative parameters were within the normal range. At 40+/-9 months after the liver transplantation a significant decrease in serum ammonia levels, AST and bilirubin and a significant increase in cholinesterase levels were observed. CONCLUSION: Patients with restored liver function after liver transplantation showed significantly improved ERG parameters. Our data suggest a recovery of the cells involved in hepatic retinopathy, including the Müller (glial) cells.

GABA(A) receptors in Müller glial cells of the human retina.

The present study was aimed at characterizing the GABA(A) receptor-mediated currents in acutely isolated glial (Müller) cells of the human retina and investigating their subcellular localization across the Müller cell membrane. Extracellular application of GABA evoked two current responses in human Müller cells: a fast transient GABA(A) receptor-mediated current that inactivated within 10 s and that was independent of extracellular Na(+), and a sustained current that was dependent on extracellular Na(+) and that was mediated by high-affinity GABA transporters. The receptor current was half-maximally activated at a GABA concentration of 32 microM, while the transporter current showed an affinity constant of 7.9 microM GABA. The receptor currents were blocked by bicuculline and picrotoxin and were also activated by muscimol or by other amino acids. The receptor currents are Cl(-) currents, as indicated by the close relationship between the reversal potential of these currents and the Cl(-) equilibrium potential. Using perforated-patch recordings, a mean intracellular Cl(-) concentration of 37 +/- 12 mM was determined in human Müller cells. Using electrophysiological and fluorescence imaging methods, it was revealed that GABA(A) receptors are unevenly distributed across the Müller cell membrane, with higher densities at the endfoot, at the soma, and at the distal sclerad end of the cells. It is concluded that GABA(A) receptor expression may allow a sensing of retinal GABAergic neuronal signal transmission by Müller cells.