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Background: Nuclear factor-kappa B (NF-kappaB) is elevated in regulating transcription of many cytokines and inflammatory mediators. The purpose of this study was to investigate the activation and the significance NF-kappaB in lipopolysaccharide (LPS) induced keratitis., Methods: LPS induced keratitis model was based on Wistar rats. At 0.5, 1, 3, 6, 12, 24 or 72 hours after LPS treatment, the rat corneas were observed with a slit lamp microscope, then the rats were sacrificed and their corneas were excised for routine histological analysis. The expression of NF-kappaB was detected with immunohistochemical staining. The change of tumour necrosis factors-alpha (TNF-alpha) mRNA expression was identified by reverse transcriptase polymerase chain reaction (RT-PCR)., Results: Histological findings demonstrated that LPS treated corneas showed significant changes in corneal structure. Corneal edema, pronounced inflammatory cells infiltration and inordinate collagen fibres were observed. Immunohistochemical results showed that the expression of NF-kappaB and its activation obviously increased after LPS treatment compared with the normal group and control group. Positive cells could be observed at 0.5 hour and peak expression of NF-kappaB was observed between 3 hours and 12 hours after infection, but returned to or approached normal level by 72 hours. RT-PCR showed that the level of TNF-alpha mRNA began to increase 0.5 hour after LPS treatment, peaked at 6 hours and then subsided by 72 hours. NF-kappaB had a positive correlation with the expression of TNF-alpha mRNA (r = 0.964, P < 0.01), both NF-kappaB and TNF-alpha had a strong positive correlation with the degree of inflammatory response in LPS treated corneas (r = 0.929, P < 0.01; r = 0.587, P < 0.05, respectively)., Conclusions: The activation of NF-kappaB was increased in LPS treated corneas and was elevated in LPS induced keratitis by promoting overexpression of TNF-alpha mRNA. NF-kappaB may play an important role in the pathogenesis of LPS-associated keratitis in rats.

The aim of the present study was to investigate the efficacy of fucoxanthin on endotoxin-induced uveitis (EIU) in rats. The effects of fucoxanthin on endotoxin-induced leucocyte and protein infiltration, nitric oxide (NO), prostaglandin (PG)-E2 and tumour necrosis factor (TNF)-alpha concentrations in rat aqueous humour, as well as on the cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) protein expression in a mouse macrophage cell line (RAW 264.7 cells) were studied. EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). Immediately after the LPS injection, either 0.1, 1 or 10mgkg(-1) of fucoxanthin was injected intravenously. The aqueous humour was collected 24hr later from both eyes, and both the number of cells infiltrating into the aqueous humour and the aqueous humour protein concentration were measured. The levels of PGE2, NO and TNF-alpha were determined by enzyme-linked immunosorbent assay. The RAW 264.7 cells were pretreated with various concentrations of fucoxanthin for 24hr and subsequently incubated with LPS for 24hr. COX-2 and iNOS protein expression was analysed by the Western blotting method. Levels of PGE2, NO and TNF-alpha production were determined. Fucoxanthin suppressed the development of EIU in a dose-dependent fashion. Treatment with fucoxanthin resulted in a reduction in PGE2, NO and TNF-alpha concentrations in the aqueous humour. The expression of COX and iNOS protein in the fucoxanthin treated RAW264.7 cells decreased significantly compared to that the LPS group. It also significantly reduced the concentration of PGE2, NO and TNF-alpha production in the medium of cells. The present result indicate fucoxanthin suppresses the inflammation of EIU by blocking the iNOS and COX-2 protein expression and its anti-inflammatory effect on eye is comparable with the effect of predinisolone used in similar doses.

Purpose: Caffeic acid phenethyl ester (CAPE) has antimicrobial, anti-inflammatory, antioxidant, immunomodulatory, and carcinostatic properties. In this study, the efficacy of CAPE in endotoxin-induced uveitis (EIU) in rats is investigated., Methods: EIU was induced by a footpad injection of lipopolysaccharide (LPS). In the treatment group, 10 micromol/kg CAPE was injected intraperitoneally immediately after LPS injection. At 24 hr after LPS injection, the number of infiltrating cells, protein concentration, and levels of myeloperoxidase (MPO) in aqueous humor; malondialdehyde (MDA), MPO, and total antioxidant levels in serum were determined. Eyes were enucleated for histopathologic evaluation, and, counting inflammatory cells in iris-ciliary body (ICB), the efficacy of treatment was determined., Results: CAPE significantly suppressed LPS-induced increase in the number of inflammatory cells (p = 0.0001), protein concentration (p = 0.0001), and MPO levels (p = 0.0001) in aqueous humor as well as MDA (p = 0.001) and MPO (p = 0.0001) levels in serum. Histopathologic evaluation of ICB showed significant reduction in the inflammatory cell counts in the treatment group (p = 0.0001)., Conclusions: CAPE was found efficient in suppressing inflammation and ocular tissue damage induced by LPS in rats.

The aim of this study was to investigate the efficacy of naringin and naringenin on endotoxin- induced uveitis (EIU) in rats. EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). The rats were injected intravenously with 0.4, 4, or 40 microg/kg naringin or naringenin. Each compound was administered three times, simultaneously, 30 min before and after the actual LPS injection. The aqueous humor was collected 24 h later from both eyes, and the number of cells infiltrating into the aqueous humor and the aqueous humor protein concentration were measured. The levels of prostaglandin E2 (PGE2) and nitric oxide (NO) were determined. Naringin and naringenin suppressed the development of EIU in a dose-dependent fashion. Both treatments with naringin and naringenin produced reductions in PGE2 and NO concentrations in the aqueous humor. In particular, 40 microM/kg of naringin and naringenin suppressed increases in cell count owing to LPS treatment by 31% and 38%, respectively. The possible mechanism for the antiocular inflammatory effect may be the suppression of PGE2 and NO by naringin and naringenin.

Aims: Diabetic patients may have abnormal inflammatory reactions to foreign or endogenous stimuli. This study was designed to evaluate inflammatory reactions in the diabetic eye through retinal leucocyte dynamics in the inflamed eyes of diabetic rats., Methods: Three weeks after diabetes induction in Long-Evans rats, endotoxin induced uveitis was produced by footpad injection of lipopolysaccharide (LPS). After LPS injection, leucocyte behaviour was evaluated in vivo by acridine orange digital fluorography., Results: The number of rolling leucocytes increased in a biphasic manner at 12 hours and 48 hours. The number of leucocytes accumulating in the retina reached a peak at 72 hours. The maximal numbers of rolling and accumulating leucocytes in the diabetic retina decreased by 56.3% (p<0.01) and 46.7% (p<0.0001), respectively, compared with the non-diabetic retina. The levels of mRNA expression of adhesion molecules in the retina, which were upregulated after LPS injection, were also lower in diabetic rats than in non-diabetic rats., Conclusion: This study is the first to show that endotoxin induced inflammation is disturbed in the diabetic eye, based on evidence that the leucocyte-endothelial cell interactions stimulated by LPS were suppressed in the diabetic retina. These findings support the theory that ocular inflammatory reactions are impaired in diabetic patients.

Purpose: To examine whether the angiotensin II type 1 receptor (AT1-R) signaling plays a role in ocular inflammation in endotoxin-induced uveitis (EIU)., Methods: EIU was induced in C57BL/6 mice by a single intraperitoneal injection of 150 mug lipopolysaccharide (LPS). Tissue localization, mRNA expression, and protein levels of AT1-R in murine retinas were examined by immunohistochemistry, RT-PCR, and Western blot analyses, respectively. Telmisartan, an AT1-R antagonist widely used as an antihypertensive agent, was administered intraperitoneally at a dose of 10 mg/kg daily for 5 days until the injection of LPS. Twenty-four hours after administration, leukocyte adhesion to the retinal vasculature was evaluated with a concanavalin A lectin perfusion-labeling technique. Retinal mRNA and protein levels of intercellular adhesion molecule (ICAM)-1 were examined by RT-PCR and ELISA, respectively. Protein concentration and inflammatory cells in the aqueous humor were also measured., Results: Retinal vessels were positive for AT1-R. In mice with EIU, retinal AT1-R mRNA and protein levels were significantly increased when compared to the normal control. EIU animals also showed significant increases in the number of inflammatory cells infiltrating the anterior chamber and adhering to the retinal vessels and in retinal ICAM-1 levels. Administration of telmisartan to EIU mice resulted in significant suppression of retinal ICAM-1 expression and leukocyte adhesion and infiltration compared with vehicle treatment. Protein concentration in the aqueous humor of telmisartan-treated EIU mice tended to be lower than that of vehicle-treated EIU mice, but the difference was not statistically significant., Conclusions: AT1-R signaling blockade inhibited retinal ICAM-1 upregulation and leukocyte adhesion and infiltration in the EIU model. These results suggest the potential use of an AT1-R antagonist as a therapeutic agent to reduce ocular inflammation.

Background: It is relatively rare to encounter a case of neovascular glaucoma induced by malignant lymphoma metastasized into the eyeball., Case: A 79-year-old woman initially visited our ophthalmology clinic with the chief complaint of blurring of vision in the left eye which was affected by neovascular glaucoma. She had a history of systemic malignant lymphoma, first diagnosed from skin biopsy and treated to complete remission 3 years previously. A metastatic brain lesion, detected 2 months before her initial visit to our clinic, was cured by radiotherapy., Findings: By ultrasound biomicroscopy (UBM) we detected abnormal thickening of the temporal half of the ciliary body of her left eye. Cytological examination of the aqueous humor revealed invasion by malignant cells of presumed lymphocyte origin. Radiation therapy to the left eye normalized the intraocular pressure in a week, followed by a reduction in the neovascularization of the iris and the thickening of the ciliary body., Conclusion: This case showed that metastatic malignant lymphoma in the eye could result in neovascular glaucoma and that UBM is useful to detect and to observe lesions in the iris and ciliary body.

There are species-related anatomical differences in the ciliary muscle of the avian eye. The arrangement of muscular fibers in the avian eye is not well defined. To clarify this situation, we studied the architecture of ciliary muscle of Gallus domesticus by light and scanning electron microscopy (SEM). Our results showed the existence of three main muscular groups that we defined as anterior, posterior, and intermediary. These muscle divisions correspond to the description of the ciliary muscle as previously stated by Crampton (1813), Brucke, and Muller (1856). The striated fibers have a meridian orientation. The anterior and posterior muscular groups are inserted in the sclera, around the Schlemm's canal wall and ciliary process stroma. The vitreal intermediary muscle has fibers inserted in Schlemm's canal wall and ciliary process stroma. The framework of these muscular fibers may according to its insertions participate in the visual accommodation mechanism and outflow of the aqueous humor system.

Antiglaucoma drugs with anti-inflammatory properties may be of particular value for the long-term treatment of glaucoma since they may reduce the risk for treatment-related inflammatory processes in outer compartments of the eye. The purpose of this study was, to evaluate the effect of systemic and topical administration of GLC756, a novel mixed dopamine D(2) receptor agonist and D(1) receptor antagonist which lowered intraocular pressure in man, and timolol on endotoxin-induced-uveitis (EIU) in rats. For EIU, 8-week-old Lewis rats received an intravenous injection of 160 microg lipopolysaccharide (LPS) from Salmonella typhimurium. GLC756, timolol, or betamethasone, as positive control, were administered either topically (0.4, 0.5, and 0.1%, respectively, 16-times 20 microl eye drops during 48 hr) or systemically (1 mg kg(-1) subcutaneously for 5 days). Protein content, released TNF-alpha, the number of cells as well as cells expressing TNF-alpha were determined in aqueous humor 48 hr after LPS-injection and served as parameters for inflammation. LPS induced an increase of protein content, infiltrating cells and cells expressing TNF-alpha in the aqueous humor. Topical and systemic administration of GLC756 and betamethasone, almost completely suppressed the increase of protein content and betamethasone in addition also suppressed the number of cells in aqueous humor. In conclusion, the almost complete suppression of LPS-induced protein increase in aqueous humor by GLC756 suggests an additional anti-inflammatory potential of dopaminergic compounds in glaucoma treatment. Timolol did not show any effect on EIU in rats.

Lymphangiogenesis, the formation of new lymphatic vessels, is important for tumor metastasis and induction of immunity to peripheral antigens including organ transplants. We herein describe a novel mouse model of spontaneous, secondary lymphangiogenesis in the normally avascular cornea. corn1 mice, which suffer from a deletion in the gene encoding the cytoskeletal protein destrin, develop hemangiogenesis as well as spontaneous outgrowth of LYVE-1+++/CD31+ lymphatic vessels into the cornea starting at age 4 weeks. Corneal lymphangiogenesis is delayed in onset, is less intense, and regresses earlier compared with hemangiogenesis. Moreover, the lymphangiogenesis is preceded only by a mild recruitment of CD45+ inflammatory cells into the cornea. In contrast to mice with inflammation-induced hem- and lymphangiogenesis, corn1 mice do not develop breakdown of the blood-aqueous barrier. Finally, in this novel mouse model, a blocking anti-VEGFR3 antibody significantly inhibited not only lymph- but also hemangiogenesis. In summary, destrin deletion has differential effects on spontaneous hem- and lymphangiogenesis in the normally avascular cornea and represents a novel mouse model to study the mechanisms of lymphangiogenesis and to test the antihem- and antilymphangiogenic properties of known or new antiangiogenic agents.

Purpose: To report acute unilateral hypopyon uveitis as an initial presenting feature of relapsing acute lymphoblastic leukemia (ALL) in an adult patient., Design: Observational case report., Methods: Anterior chamber paracentesis was performed in a 56-year-old male presenting with treatment-resistant unilateral hypopyon while in the remission phase of ALL., Results: Examination of the aqueous humor aspirate revealed presence of malignant cells compatible with the previous bone marrow biopsy and subsequent spinal tap results., Conclusions: Atypical hypopyon uveitis can be an indication of relapsing ALL, even in adults. Prompt anterior chamber aspiration is required for the correct diagnosis and subsequent treatment.

Purpose: Anterior chamber involvement is unusual in patients with acute lymphoblastic leukemia (ALL) and has never been described in the setting of Ph+ (Philadelphia chromosome-positive) ALL. Moreover, there have been no reports of this complication as a primary relapse in a patient treated with imatinib., Design: Interventional case report., Methods: A 55-year-old woman with Ph+ ALL in complete remission with imatinib and presenting unilateral anterior uveitis at initial examination was clinically evaluated. Hematologic and ocular studies were performed: blood films, bone marrow smears, and anterior chamber paracentesis with aqueous fluid cytology., Results: Although there was no evidence of leukemia in the blood or bone marrow samples, aqueous fluid cytology identified Ph+ positive lymphoblastic leukemic cells., Conclusions: The patient had developed anterior chamber infiltration without hematological relapse while treated with imatinib. In our opinion, paracentesis should be performed without delay when uveitis develops in ALL, regardless of systemic relapse.

Purpose: To determine the effect of pyrrolidine dithiocarbamate (PDTC), a nuclear factor (NF)-kappaB inhibitor, on chemokine and chemokine receptor expression and thus elucidate the role of NF-kappaB in the pathogenesis of experimental autoimmune anterior uveitis (EAAU)., Methods: Uveitis was induced in Lewis rats with the injection of melanin-associated antigen into the footpad. PDTC (200 mg/kg and 100 mg/kg) was administered intraperitoneally daily, beginning 1 day after the immunization. The clinical inflammatory activity of the anterior chamber was recorded daily and scored. Immunohistochemical staining and an electrophoretic mobility shift assay assessed the effect of PDTC on NF-kappaB activation in the iris/ciliary body tissues. Gene expression profiles of chemokine and chemokine receptors were semiquantitatively examined by reverse transcriptase-polymerase chain reaction (RT-PCR). Aqueous chemokine levels were measured by enzyme-linked immunosorbent assay (ELISA)., Results: PDTC significantly attenuated the clinical scores and monocyte/lymphocyte infiltration in rats with EAAU. PDTC effectively inhibited NF-kappaB activation in the iris and ciliary body, and markedly inhibited the expression of chemokine genes, including monocyte chemoattractant protein (MCP)-1, regulated-on-activation normal T-cell expressed and secreted (RANTES), and interleukin (IL)-8 and chemokine receptors genes including CCR2, CCR5, and CXCR3., Conclusions: Activation of NF-kappaB appears to play an important role in the pathogenesis of EAAU, through transcriptional control of MCP-1, RANTES, and IL-8 gene expression. Blocking NF-kappaB reduces ocular inflammation and may be an effective strategy in the treatment of acute anterior uveitis.

Aims: The aim of this study was to investigate the effects of pyrrolidine dithiocarbamate (PDTC), a nuclear factor (NF)-kappaB inhibitor, on cytokine expression and suppression of anterior chamber inflammation in experimental autoimmune anterior uveitis. Uveitis was induced in the Lewis rats with the injection of a melanin-associated antigen into the peritoneum and footpad. At defined time points, cytokine mRNA expressions in the iris and ciliary body were measured by using a semiquantitative polymerase chain-reaction method., Results: We found that interferon-gamma (IFN-gamma) and tumor necrosis factor (TNF)-alpha mRNA expression peaked during the active phase of uveitis, whereas interleukin (IL)-10 mRNA increased during the disease resolution. In a separate experiment, PDTC (100 and 200 mg/kg/day) was administrated intraperitoneally daily after immunization. We found that PDTC (100 and 200 mg/kg/day) effectively suppressed ocular inflammation, as indicated by reduced clinical scores and inflammatory cells infiltration in aqueous humor and the iris and ciliary body., Conclusions: The inhibitory effects of PDTC are mainly resulted from inhibiting the expression of proinflammatory cytokines, TNF-alpha and IFN-gamma but augmenting anti-inflammatory cytokines, IL-10 expression. These findings suggest that the application of NF-kappaB inhibitors may be a potential therapeutic method for the treatment of acute anterior uveitis.

The purposes of this study are to determine the genes that are up- or down-regulated in eyes with endotoxin-induced uveitis (EIU) by an oligonucleotide microarray system, and to determine the temporal and spatial changes in expression of selected genes that show strong up-regulation. EIU was induced by a footpad injection of lipopolysaccharide (LPS) in male Lewis rats. The expression of genes in the iris-ciliary body (ICB) at 2, 6, 12, and 24 hr after LPS injection was determined by oligonucleotide microarray analyses and compared to that in control rats. The microarray displayed 9911 genes and expressed sequence tags (ESTs). Cluster analysis was performed for highly up-regulated genes. Selected genes for cytokines (interleukin (IL)-1 beta and IL-6), chemokines (RANTES), and immediate early genes (Jun B, c-Fos, and c-Jun) were also studied by real-time polymerase chain reaction (PCR). Immunohistochemical studies were performed to localize the protein expression of some immediate early gene products. After LPS injection, the expression of 1930 genes were increased or decreased over 2-folds compared with normal controls by 24 hr. One hundred and seventeen genes were up-regulated over 10-fold, and these were classified into five clusters with similar expression pattern. The immediate early genes and transcription factors genes were included in one cluster of up-regulated genes peaking at 2 hr after the LPS injection. The expressions of cytokines, chemokines, and adhesion molecules were highly up-regulated. Real-time PCR analyses for selected genes showed similar expression changes as detected by the microarray analyses. Jun B immunoreactivity was found in the ICB cells at 3 and 6 hr after LPS injection. Gene expression changes after LPS injection were profiled by using an oligonucleotide microarray system. Our data suggest that the immediate early genes, such as Jun B, play an important role in inducing the inflammatory-related genes in the ICB.

Diagnosis and management of intraocular inflammation involves the assessment of cells and protein levels ("flare") in the aqueous humor. These factors are difficult to quantify precisely on clinical examination alone. Laser flare-cell photometry provides an automated technique to quantify these factors objectively, and it has been used in a variety of research and clinical situations to assess anterior segment inflammation. Any new technique requires evaluation to determine accuracy and reproducibility of measured values, and initial applications require critical appraisal to assess the value of the technique. Both in vitro and in vivo studies of laser flare-cell photometry have been performed to determine its validity and utility as a research and clinical tool. This article reviews published studies that describe the technique of laser flare-cell photometry; it provides new in vitro data that supplements information on the capabilities of this technique and factors that influence photometry results, and it reviews representative publications that have used laser flare-cell photometry for study of specific disease entities. This information can help clinicians and researchers to become familiar with the strengths and limitations of laser flare-cell photometry, to identify appropriate future uses for this technique, and to use it and interpret its results appropriately. Laser flare-cell photometry offers an opportunity to improve upon current techniques of inflammation assessment and should not be considered simply an objective surrogate for clinical grading of cells and flare at the slit-lamp biomicroscope. Its research applications and utility for monitoring patients with uveitis have not yet been fully explored.

Purpose: To quantify the breakdown of the blood-aqueous barrier (BAB) following phacoemulsification with posterior chamber lens implantation in eyes with Fuchs' heterochromic uveitis (FHU)., Patients and Methods: In this retrospective study, 19 eyes of 19 patients with FHU (mean age 38 +/- 14 years) and 35 eyes of 35 patients with senile cataracts (mean age 63 +/- 9 years) underwent phacoemulsification with one-piece PMMA posterior chamber lens implantation. Aqueous flare was quantified using the laser flare-cell meter (LFCM, Kowa FC-1000) following medical pupillary dilation preoperatively and 1, 3, and 5 days, then 1 and 6 weeks and 6 months postoperatively., Results: Mean preoperative aqueous flare (in photon counts per millisecond) in FHU vs. controls was 11.7 +/- 3.5 vs. 5.8 +/- 1.7. Following cataract surgery, mean aqueous flare increased to 27.8 +/- 4.4 vs. 16.0 +/- 4.5 on day 1, decreased to 23.6 +/- 4.0 vs. 11.8 +/- 3.5 on day 3, and to 18.0 +/- 3.0 vs. 9.5 +/- 1.7 on day 5. In FHU eyes, it was 13.9 +/- 2.7 after 1 week, and had returned to preoperative levels after 6 weeks (10.9 +/- 2.5) and remained stable for up to 6 months (mean 10.3 +/- 2.2). Pre- and postoperatively, aqueous flare values were 2-3 times higher in FHU eyes than in control eyes with senile cataract (p = 0.01). No postoperative complications such as fibrin formation, synechiae, macrophages on the intraocular lens optic or endophthalmitis were observed in any of the patients., Conclusions: BAB breakdown following phacoemulsification with posterior chamber lens implantation is relatively mild in eyes with FHU and the BAB appears to be fully reestablished to preoperative levels 6 weeks postoperatively, explaining the usually good outcome of cataract surgery in this condition., (Copyright (c) 2005 S. Karger AG, Basel.)

Purpose: Aronia crude extract (ACE) with high levels of polyphenol compounds has been reported to have antioxidative effects in vitro and in vivo. In this study, attention was focused on the antioxidant effect of ACE. The purpose of the present study was to investigate the effect of ACE on endotoxin-induced uveitis (EIU) in rats. In addition, the endotoxin-induced expression of the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 proteins was investigated in a mouse macrophage cell line (RAW 264.7) treated with ACE in vitro, to clarify the anti-inflammatory effect., Methods: EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). Immediately after the LPS inoculation, 1, 10, or 100 mg ACE or 10 mg prednisolone was injected intravenously. After 24 hours, the aqueous humor was collected from both eyes, and the number of infiltrating cells, protein concentration, nitric oxide (NO), prostaglandin (PG)-E2, and TNF-alpha levels in the aqueous humor were determined. RAW 264.7 cells treated with various concentrations of ACE were incubated with 10 mug/mL LPS for 24 hours. Levels of NO, PGE2, and TNF-alpha were determined by an enzyme-linked immunosorbent assay. The expression of iNOS and COX-2 proteins was analyzed by Western blot analysis., Results: The number of inflammatory cells, the protein concentrations, and the levels of NO, PGE2, and TNF-alpha in the aqueous humor in the groups treated with ACE were significantly decreased in a dose-dependent manner. In addition, the anti-inflammatory effect of 100 mg ACE was as strong as that of 10 mg prednisolone. The anti-inflammatory action of ACE was stronger than that of either quercetin or anthocyanin administered alone. ACE also suppressed LPS-induced iNOS and COX-2 protein expressions in RAW 264.7 cells in vitro in a dose-dependent manner., Conclusions: The results suggest that ACE has a dose-dependent anti-ocular inflammatory effect that is due to the direct blocking of the expression of the iNOS and COX-2 enzymes and leads to the suppression of the production of NO, PGE2, and TNF-alpha.

Purpose: To evaluate biocompatibility of Carbon (C) and Titanum (T) surface modified Polymethyl-methacrylate (PMMA) Intraocular Lens (IOLs) in pseudophakic rabbit model., Methods: Extracapsule Cataract Extraction (ECCE) and IOLs implantation were performed in Japanese albino rabbit eyes. The white cell concentration from the aqueous humor in the eyes was counted within 3 months post operation. The inflammatory cells in the eyes were noted and graded by slit lamp using a semiquantitive scale up to 1-year post operation. The rabbit eyes were inspected under light microscopy, where pathology of the eyes was caracterized., Results: The white cell concentration in the aqueous humor was significantly attenuated in C and T IOL-implanted eyes compared with PMMA IOL-implanted eyes 1 week post operation. The exudate levels in the anterior ocular chamber and the posterior synechias were significant lower in T IOL-implanted eyes than in PMMA IOL-implanted eyes 1 week and 2 weeks after surgery. The exudate levels in the anterior chambers and the posterior synechias were not significantly different in C IOL-implanted versus PMMA IOL-implanted eyes., Conclusions: This in vivo study provides evidence of effectiveness of Carbon and Titanium IOLs in improving the biocompatibility of PMMA IOLs.

Purpose: To study, with an objective method, inflammation of the anterior segment of the glaucomatous eye after treatment with latanoprost, travoprost and bimatoprost., Materials and Methods: Sixty patients with chronic open-angle glaucoma aged between 38 and 76 years (mean 64.0 +/- 12.2) were randomly assigned to latanoprost 0.005, travoprost 0.004 and bimatoprost 0.03%. The study period lasted 6 months. Intraocular pressure (IOP) was measured every 2 weeks. We studied the intraocular inflammation before and after 3 and 6 months of therapy with an instrument composed of a He-Ne laser beam system, a photomultiplier mounted on a slitlamp microscope and a computer. This flare meter allows objective determination of the flare and the number of cells in the aqueous of the anterior chamber., Results: At the baseline, IOP was 26.4 +/- 3.6 mm Hg. After 3 months of treatment, mean IOP in the latanoprost group was 17.9 +/- 0.3 mm Hg (p < 0.001) with a mean cellularity of 12.638 +/- 3.284 photons/ms (p < 0.001). The travoprost group had an IOP of 17.2 +/- 0.3 mm Hg (p < 0.001) with a cellularity of 9.719 +/- 1.927 photons/ms (0.001). Finally, IOP in the bimatoprost group was 17.6 +/- 0.5 mm Hg (p < 0.001) with a cellularity of 6.138 +/- 1.475 photons/ms (p < 0.032). After 6 months of treatment, IOP in the latanoprost group was 18.1 +/- 0.3 (p < 0.001), in the travoprost group 17.3 +/- 0.3 (p < 0.001) and in the bimatoprost group 17.7 +/- 0.5 mm Hg (p < 0.001), whereas cellularity was 11.838 +/- 3.218 (p < 0.001), 8.950 +/- 3.692 (p < 0.001) and 7.617 +/- 2.603 photons/ms (p < 0.001), respectively. After 3 months, the travoprost (p < 0.013) and the bimatoprost groups (p < 0.001) had less flare compared with the latanoprost group and this remained so even at 6 months. When we compared the travoprost group with the bimatoprost group, we found significantly less flare at 3 months in the bimatoprost group (p < 0.001) but not at 6 months (p < 0.246)., Conclusions: The flare meter analysis shows that the eyes treated with bimatoprost and travoprost have a less significantly broken blood-aqueous barrier and their anterior chamber is also significantly less inflamed., (Copyright 2004 S. Karger AG, Basel)

Background: Cytology of the aqueous humor can confirm the diagnosis of many intraocular conditions by revealing inflammatory cells, ghost red cells, lens material, and microorganisms., Purpose: The purpose of this study was to emphasize the contribution of aqueous humor fine needle aspiration cytology in cases where there is a clinical diagnostic dilemma whether an infectious or a noninfectious inflammatory process of the uvea is present., Methods: Collection of 0.15-0.2 ml aqueous humor was carried out with an insulin needle. The aspirated aqueous was then prepared for examination using either the Cytospin or the Thin-Prep technique and smears were stained with modified Papanikolaou and May-Grünwald-Giemsa stains., Results: Cytology of the aqueous humor essentially facilitated the differential diagnosis in 16 cases with an anterior uveal inflammation clinical picture. The cases included phacoanaphylactic endophthalmitis (n = 6), chronic postoperative bacterial endophthalmitis (n = 3), phacolytic glaucoma (n = 2), ghost cell glaucoma (n = 2), metastatic endophthalmitis (n = 1), post-traumatic lenticular abscess (n = 1), and iridocyclitis (n = 1). Treatment of the cases was successful., Conclusions: In selected cases, cytology of the aqueous humor reveals specific, with regard to the clinical entity, cell populations and other elements and thus contributes effectively to the differential diagnosis and therapeutic approach to some anterior uvea inflammations.

Objective: To quantify aqueous flare and cells in the eyes of patients with inflammation of anterior uvea by FC-2000 laser flare cell meter (LFCM), and to compare these results with those obtained with slit lamp microscopy., Methods: Aqueous flare and cells of 194 eyes of 110 patients with inflammation of anterior uvea and 52 eyes of 52 healthy subjects were graded into 0, 1+, 2+, 3+ and 4 + scale based on a previously described system using slit lamp microscopy. LFCM was also used for evaluation of aqueous flare and cells., Results: All eyes in normal individuals were graded as "0" scale of both flare and cells by silt lamp microscopy. Flare of grade 0, 1+ and 2+ were noted in 74, 98 and 18 eyes, and cell of grade 0, 1+, 2+, 3+ and 4+ were noted in 124, 26, 19, 14 and 11 eyes in uveitis patients, respectively. LFCM examination revealed that the mean flare values in uveitis eyes with flare of grade 0, 1+ and 2+ were 7.9, 29.5 and 189.0 photon count/ms, respectively. In patients with flare of grade 3+ and 4+, LFCM readings were unreliable because of increased background noise. There was significant correlation between slit lamp examination and the laser flare measurement for flare of grade 0, 1+ and 2+ (r = 0.75, P < 0.001). The mean flare values were significantly higher in patients with flare of grade 0, 1+ and 2+ than that in normal controls (5.3 pc/ms) (t = 5.872, P < 0.05). The mean cell numbers in the eyes with cell of grade 0, 1+, 2+, 3+ and 4+ were 1.5, 12.1, 33.9, 84.9 and 193.1 count/0.5 mm(3), respectively. The results of slit lamp examination showed significant correlation with laser cell counts measurement (r = 0.72, P < 0.001). The mean cell numbers were significantly higher in uveitis patients than that in normal controls (0.9 count/0.5 mm(3)) (t = 7.351, P < 0.05)., Conclusions: Our results indicate that LFCM is able to evaluate precisely the mild and moderate breakdown of blood aqueous barrier and inflammation of the anterior uvea tract, therefore it provides an important parameter for the treatment of anterior uveitis.

Purpose: Ginkgo biloba extract (GBE) contains many different flavone glycosides and terpenoides. Several previous studies have demonstrated that GBE exhibits a wide variety of biological activities, including an antioxidant action, on which we focused our attention. The aim of the present study was to investigate the efficacy of GBE on endotoxin induced uveitis in rats. The anti-inflammatory potency of GBE in vivo was compared with that of prednisolone. In addition, we also investigated nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-alpha (TNF-alpha) and the expression of iNOS in a mouse macrophage cell line (RAW 264.7) treated with GBE in vitro to clarify the anti-inflammatory effect., Methods: EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). Immediately after the LPS inoculation, either 1, 10 or 100 microg of GBE were injected intravenously. 24hr later, the aqueous humor was collected from both eyes, and the number of infiltrating cells, protein concentration and NO level in the aqueous humor was determined. The RAW 264.7 cells were pretreated with various concentrations of GBE for 24hr and subsequently incubated with LPS for 24hr. Levels of NO, PGE2 and TNF-alpha were determined by enzyme-linked immunosorbent assay. The expression of iNOS protein was analyzed by Western blotting method., Results: GBE treatment in vivo decreased the concentrations of protein and NO in the aqueous humor of EIU rats. The anti-inflammatory effect of 1 mg GBE was as strong as that of same dose prednisolone. It also significantly reduced the concentration of PGE2, TNF-alpha and NO production in the medium of RAW 264.7 cells compared to that of the LPS group in vitro. The expression of iNOS protein in the 1000 microg ml(-1) of GBE treated cells decreased significantly., Conclusion: The present results indicate GBE suppresses the inflammation of EIU by blocking the iNOS protein expression and its anti-inflammatory effect on eye is comparable with the effect of prednisolone used in similar doses.

Background: The exact mechanism of human corneal allograft rejection, which is the major cause of corneal transplant failure, remains unclear. We investigated the role of natural killer (NK) cells in rat corneal allograft rejection by examining the aqueous humor (AH) cell infiltrate on different postoperative days., Methods: Flow cytometric analysis was performed on the AH and submandibular draining lymph node (DLN) cells before transplantation and at different time points thereafter. In addition, we performed functional cytotoxicity assays with cells present in the AH during corneal rejection., Results: We demonstrated a gradual increase in the absolute cell number of different hematopoietic subpopulations in the AH after allogeneic cornea transplantation. CD3CD4 cells, mainly monocytes and macrophages, were the predominant subpopulation 2 days after transplantation, followed by a successive relative increase of CD4 T cells, CD8 T cells, CD161 T cells, and NK cells. NK and CD161 T cells were present at a 10- to 15-fold higher percentage than in the DLN, suggestive of local expansion of these cells. A higher percentage of NK cells were CD8-negative compared with DLN NK cells. AH cells specifically lysed allogeneic cells, and this cytotoxicity was mainly attributable to NK cells but not to CD4 or CD8 T lymphocytes., Conclusion: These results confirm the crucial role of CD4 cells in the allogeneic corneal graft rejection process and implicate NK cells as possible mediators of the rejection.

Purpose: To compare the postoperative inflammatory reaction induced by laser photolysis and conventional phacoemulification., Methods: Anterior chamber flare and cells were measured preoperatively and 1 day, 1 week, 1 month and 3 months postoperatively in two groups. In group 1, thirty-two eyes underwent laser photolysis. In group 2, thirty-five eyes underwent conventional phacoemulification., Results: Anterior chamber flare and cell measurements had no significant difference between two groups preoperatively and 1 day, 1 week, 1 month and 3 months postoperatively. Flare and cell values returned to preoperative values by the third month., Conclusions: Postoperative inflammatory response to laser photolysis was equal to conventional phacoemulsification, and recovery occurred in 3 months postoperatively.

Purpose: To evaluate the influence of peripheral 360 degrees retinal cryocoagulation on the blood-aqueous barrier of patients with retinal vascular disorders., Methods: The aqueous of 50 eyes of 50 patients with diabetic retinopathy or central vein occlusion was measured by laser flare photometry (FC-2000, Kowa) before peripheral 360 degrees retinal cryocoagulation and 1 day, 1 week, and 1 month thereafter., Results: Mean aqueous flare values before treatment were 15.7 (+/-8.2) photon counts/ms; mean cell count was 9.8 (+/-14.8) cells/0.5 mm(3). One day after retinal cryocoagulation flare values had increased statistically significantly to a mean of 39.2 (+/-85.8) photon counts/ms, while the increase in mean cell number to 15.0 (+/-37.2) cells/0.5 mm(3) did not reach statistical significance. One week following treatment, mean flare values had dropped to 30.9 (+/-49.6) photon counts/ms and were no longer significantly elevated compared with baseline, while the mean cell count of 11.1 (+/-27.0) cells/0.5 mm(3) was now statistically significantly elevated compared with baseline. One month after treatment the flare values had decreased to a mean of 19.7 (+/-12.0) photon counts/ms, and mean cell count had decreased to 8.1 (+/-10.4) cells/0.5 mm(3); at this point neither parameter showed a statistically significant difference from baseline values., Conclusion: Peripheral 360 degrees retinocryocoagulation does not lead to permanent disturbance of the blood-aqueous barrier.

Purpose: To evaluate the efficacy of a biodegradable polymeric scleral plug containing the immunosuppressive agent, FK506, in a rabbit model for experimental uveitis., Methods: The scleral plugs were prepared by dissolving poly(DL-lactide-co-glycolide; PLGA) and FK506 (weight, 8.5 mg; length, 5 mm; 1% FK506). The release of FK506 was evaluated in vitro by spectrophotometry on days 1, 3, 7, 14, 21, and 35. In vivo, FK506 concentrations of the vitreous were measured by high performance liquid chromatography 2 and 4 weeks after intravitreous plug implantation in pigmented rabbits. Sixteen pigmented rabbits were immunized twice subcutaneously with 10 mg of Mycobacterium tuberculosis H37Ra antigen. Twelve days later, the right eyes of all rabbits were challenged with an intravitreal injection of 50 micro g of antigen. After the first challenge, the 16 eyes of 16 pigmented rabbits were divided into two groups. Scleral plugs were implanted into the vitreous of the right eye of eight rabbits. Eight control rabbits received a sham device. The aqueous protein concentrations and cell counts were determined on postchallenge days 7, 14, and 28. To simulated chronic inflammation, the eyes were rechallenged with intravitreal antigen on day 14 and were observed for 1 month. Inflammation of the anterior chamber and the vitreous were graded clinically by two masked observers. Retinal function was evaluated by electroretinography (ERG) and histologic examination., Results: Clinical scores (anterior chamber cells, flare, and vitreous opacity) showed that treated eyes had significantly less inflammation than untreated eyes (P<0.001). Quantitative analysis of inflammatory cells (P<0.001) and protein concentrations (P<0.0001) in the anterior chamber showed significant decreases in treated eyes. Histopathologic examination showed marked inflammation and tissue disorganization in the untreated eyes. No retinal toxicity was detected, histopathologically and electroretinographically. After antigen rechallenge, inflammation in experimental eyes was still less than in control eyes., Conclusions: Intravitreal sustained-release of FK506 from a biodegradable polymeric scleral plug was highly effective in suppressing the inflammation of experimental uveitis in a rabbit model for at least 6 weeks. This device may be useful in the management of patients with severe chronic uveitis.

Purpose: To evaluate the time course of blood-aqueous barrier (BAB) disturbance in the early period after small-incision cataract surgery., Setting: Department of Ophthalmology, Vienna University, Vienna, Austria., Methods: In a prospective study, 15 eyes of 15 patients with age-related cataract had small-incision cataract surgery by phacoemulsification with intraocular lens implantation. Care was taken to minimize trauma to the uvea during surgery. Postoperative inflammation was assessed by measuring aqueous flare and cell count with a laser flare-cell meter. Postoperative measurements were performed hourly for the first 6 hours, every 2 hours until 12 hours, every 4 hours until 40 hours, and every 8 hours until 56 hours., Results: The time course of aqueous flare and cell count differed significantly among patients. The peak inflammatory response in most cases was 1 hour after surgery, with the response decreasing thereafter. The pattern of the time course was classified into subgroups defined by the presence and size of an initial spike immediately after surgery and the intensity of the subsequent inflammatory reaction. A slight increase in flare and cells was seen in the morning hours of the first postoperative day., Conclusions: Acute BAB disturbance within the first 48 hours after small-incision cataract surgery showed high interpatient variability. However, many differences were not detectable 1 day after surgery.

Purpose: Human cationic antimicrobial protein 18 (hCAP18, 18 kDa) was originally identified in leukocytes on the basis of its antimicrobial activity. The peptide composed of the 27 C-terminal amino acids of hCAP18 (hCAP18(109-135)) binds lipopolysaccharide (LPS). The purpose of the present study was to investigate the effects of hCAP18 peptide on endotoxin-induced uveitis (EIU) in rats., Methods: EIU was induced by footpad injection of LPS. Each rat was injected intravenously with 1, 10, or 100 micro g hCAP18 peptide in 0.1 mL of PBS immediately after LPS injection in male Lewis rats. At 24 hours after LPS injection, enzyme-linked immunosorbent assay was performed to evaluate concentrations of protein, nitric oxide (NO), tumor necrosis factor (TNF)-alpha, prostaglandin (PG)-E2, interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1 and macrophage inflammatory protein (MIP)-2 in aqueous humor. Also, EIU was evaluated by counting inflammatory cells in aqueous humor., Results: hCAP18 peptide at 10 and 100 micro g significantly suppressed an LPS-induced increase in the number of inflammatory cells and the levels of protein, NO, TNF-alpha, PGE2, MCP-1, and MIP-2. The anti-inflammatory effect of 10 micro g hCAP18 peptide was as strong as that of 100 micro g hCAP18 peptide. Treatment with 1 micro g hCAP18 peptide did not suppress EIU, compared with the LPS group., Conclusions: The present results indicate that hCAP18 peptide suppresses development of EIU. A possible mechanism for the ocular anti-inflammatory effect of hCAP18 peptide is that it suppresses onset of LPS-triggered inflammatory reactions by binding directly to LPS.

Objective: West Nile virus (WNV) disease is a zoonotic infection with recent outbreaks in the United States. Recent reports have highlighted the intraocular findings associated with WNV disease. We describe the intraocular findings observed in two patients infected by the West Nile virus., Design: Observational case reports., Methods: During an outbreak of WNV disease in Southwest Ohio, two patients with an acute onset of a systemic febrile illness accompanied by myalgia, arthralgia, headache, and a maculopapular rash were referred for blurred vision. Complete ophthalmologic examination, fundus photographs, and fluorescein angiograms were obtained on both patients. Both patients underwent serologic testing for viruses and cultures for bacteria, viruses, and fungi., Results: Ophthalmologic examination in each patient revealed anterior segment and vitreous inflammatory cells and multiple partially atrophic and partially pigmented chorioretinal lesions clustered in the peripheral fundus. Fundus examination in case 2 also revealed mild disc edema in both eyes. Intracranial pressure as measured by lumbar puncture was borderline elevated. The chorioretinal lesions in both patients showed a striking similarity and appeared hypofluorescent centrally and hyperfluorescent around the edges on a fluorescein angiogram. Serologic testing for the WNV was positive in both patients, and tests for all other bacteria, fungi, and viruses were negative., Conclusions: WNV usually causes mild symptoms, but it occasionally causes neurologic illness with fatal outcome or severe morbidity. We present the cases of two patients with serology-proven WNV disease who developed chorioretinal lesions with a targetlike appearance and iridocyclitis.

Purpose: To investigate possible relationships between laser flare photometry values, complications of uveitis, and outcomes in children with chronic uveitis., Design: Retrospective chart review., Method: We evaluated patients with active, noninfectious anterior, intermediate, or panuveitis who were 16 years of age or younger and who had laser flare photometry at one of two academic institutions. Complications enumerated at baseline and during follow-up were compared with laser flare photometry values and to anterior chamber cell counts., Results: At least one laser flare photometry value ("flare"), defined as baseline measurement, was available for 59 patients (41 girls, 18 boys; mean age, 10.3 +/- 3.5 years); 38 of these patients had at least one additional measurement during follow-up (median 11 months). Complications of uveitis were present in 35 patients (59%) at baseline. There was a positive association between increased laser flare photometry values and complications at baseline (any complication [P =.007], posterior synechiae [P =.003]). The development of complications during follow-up was associated with the presence of complications at baseline (P =.018). A subgroup of patients with low flare at baseline had no complications during follow-up regardless of treatment given., Conclusions: There is a positive relationship between laser flare photometry values and the prevalence of complications of uveitis in children. Laser flare photometry provides a novel way to monitor children with uveitis. Future study will be needed to determine whether values have prognostic importance and whether a treatment strategy that minimizes flare results in fewer uveitic complications.

Background and Objective: Corticosteroids remain the mainstay for control of ocular inflammation after vitreous surgery. A controlled, randomized, prospective study was performed to evaluate the effectiveness of a single intravitreal injection of dexamethasone phosphate on postoperative inflammation after simple vitreous surgery in patients with proliferative diabetic retinopathy and macular pucker., Patients and Methods: Aqueous cell flare intensity was measured preoperatively and on days 1, 10, and 90 in 56 consecutive patients who underwent vitreous surgery for proliferative diabetic retinopathy and macular pucker. Subjects were consecutively randomized to two groups: 400 microg of intravitreal dexamethasone (treatment group) or no dexamethasone (control group), Results: Before surgery, cell and flare intensity was similar in both groups. Flare intensity was significandy lower at 10, 30, and 90 days in the proliferative diabetic retinopathy treatment group (P < .05)., Conclusion: Intravitreal dexamethasone significantly alleviates postoperative inflammation after vitreous surgery and is a useful adjunct.

Purpose: To establish, in the scarified mouse eye, a new model of Staphylococcus aureus keratitis suitable for studies of pathogenesis and host defense mechanisms., Methods: Corneas of three strains of mice (BALB/c, A/J, and C57BL/6) were scarified and inoculated with S. aureus strain 8325-4. Mice underwent slit lamp examination (SLE) at 1, 3, 5, 7, and 9 days after infection and were killed. Histopathologic analyses, determination of bacterial colony-forming units (CFU), and myeloperoxidase (MPO) activity assays were performed at each time point., Results: S. aureus keratitis developed in both BALB/c and A/J strains of mice, but not in C57BL/6. The BALB/c and A/J strains demonstrated greater susceptibility to infection, as evidenced by significantly higher SLE scores and more viable bacteria per infected eye than in C57BL/6 mice at 5, 7, and 9 days after infection (P

The clinicopathologic correlation of two patients with primary intraocular lymphoma is described, both of whom had a rare and unusual presentation of hypopyon uveitis. Sampling of aqueous fluid proved the cytopathologic diagnosis of B-cell lymphoma in the first patient. In the second patient an iris biopsy confirmed a T-cell lymphoma. The samples were examined using haematoxylin and eosin, and immunohistochemical stains. These patients were diagnosed with primary intraocular lymphoma on initial presentation with hypopyon uveitis. Secondary glaucoma was noted in the patient with T-cell lymphoma. Both cases appeared confined to the eye and initially responded favourably to aggressive chemotherapy and radiotherapy, but later went on to develop systemic involvement. Clinicians should be aware of this dramatic mode of presentation, which is unusual for ocular lymphoma.

Purpose: To report a case of uveitis-glaucoma-hyphema (UGH) syndrome in which anterior chamber paracentesis led to the diagnosis of sickle cell trait., Design: Observational case report., Methods: A 43-year-old Cuban pseudophakic male was seen multiple times over a 3-year period complaining of floaters and blurry vision in his left eye. He was noted to have an inferotemporally displaced posterior chamber intraocular lens and recurrent microhyphemas with elevated intraocular pressure (IOP) readings between 29 and 46 mm Hg with each episode. He was diagnosed with UGH syndrome. Posterior chamber intraocular lens explantation and anterior chamber washout was performed. The aqueous fluid was submitted for cytopathologic examination., Results: Postoperatively, the patient's symptoms resolved and he had no further hemorrhages or elevated IOP readings. Cytopathology of the aspirate revealed sickled red blood cells., Conclusions: Microscopic examination of aqueous fluid can be a valuable tool in diagnosing ophthalmic manifestations of systemic disease.

Objective: To assess the effect of age and pupillary dilation on aqueous flare., Methods: In this study, 100 eyes of 100 patients ranging in ages from 23 to 84 years were examined. Anterior chamber flare was measured before and after pupillary dilation using the Kowa laser flare meter (FM-500). Predilation and postdilation flare counts were compared by paired t-test. Stepwise regression analysis was then used to determine the effect of demographic variables on pre- and postdilation flare as well as the difference between pre-and postdilation flare counts., Results: The predilation and postdilation flare counts correlated with age (P < 0.0001 for both pre-and postdilation flare counts). Correlation coefficient between age and flare measurements was R2 = 0.58 predilation and 0.63 postdilation. Flare intensity significantly decreased after pupillary dilation (P < 0.001)., Conclusions: Anterior chamber flare increases with age. It might be related to blood-aqueous barrier instability. Pupillary dilation significantly decreases flare counts suggesting that aqueous protein concentration is dependent on aqueous flow rates.

Purpose: To investigate the ocular pharmacodynamic profile of a polymer nanoparticle system loaded with sodium ibuprofen (IBU-RS) in comparison to an aqueous solution of ibuprofen lysinate (IBL) in the rabbit eye both being applied topically., Methods: Ocular inflammation was elicited by topical application of sodium arachidonate. Inflammation was quantified according to a modified Draize test. The protein level and the number of polymorphonuclear leukocytes in the aqueous humor were assessed after 2 h from arachidonate instillation. The ibuprofen concentration in the aqueous humor was evaluated by HPLC assay. The physico-chemical properties of nanoparticles were also evaluated., Results: The IBU-RS nanosuspension formulation significantly reduced the primary signs of ocular inflammation as well as significantly reducing the protein level and the number of polymorphonuclear leukocytes in the aqueous humor compared with the IBL formulation. Furthermore, the aqueous humor drug concentration from the group treated with IBU-RS was significantly higher compared to the IBL-treated group. The IBU-RS nanosuspensions showed very interesting size and surface charge values, adequate for ophthalmic administration., Conclusions: The pharmacological profile of the topical IBU-RS nanosuspension formulation described in this study indicates that the dispersion of the drug within RS polymer nanoparticles increased its ocular bioavailability and ultimately its pharmacological activity., (Copyright 2002 S. Karger AG, Basel)

Purpose: To determine the prevalence of detection of ghost cells (GCs) in vitreous hemorrhages (VHs) and correlate it with the development of ghost cell glaucoma. To compare our results with those previously reported., Methods: Retrospective analysis of clinical records and vitreous cytology reports (cytospin and direct paraffin embedding) from patients undergoing vitrectomy for VH at Instituto Universitario de Oftalmobiología Aplicada and University Hospital, Valladolid, for the last 2 years., Results: 76 VHs have been analysed; 45 corresponded to proliferative diabetic retinopathy and the remainder to several pathologies. GCs have been detected in 18 cases (25%) (4 out of 76 specimens were excluded): 14 were phakic and 4 pseudophakic. Most of 76 VHs lasted more than 2 months. No case of ghost cell glaucoma was detected, neither previously nor after vitrectomy., Conclusions: The frequency of detection of GCs in vitreous cytology may vary according to the cytologic technique. In contrast with the information previously reported, ghost cell glaucoma is an unfrequent disease, particularly after vitrectomy, probably because of a better removal of hemorrhagic debris from the vitreous cavity nowadays. Nevertheless, this diagnosis should not be overlooked.

Purpose: To compare the course of inflammation after small-incision cataract surgery with implantation of 1 of 3 types of foldable intraocular lenses (IOLs) in eyes with uveitis., Setting: Department of Ophthalmology, University of Vienna, Vienna, Austria., Methods: Seventy-four eyes with uveitis and cataract and 68 control eyes with cataract were prospectively selected to receive a foldable hydrophilic acrylic (Hydroview, Bausch & Lomb), hydrophobic acrylic (AcrySof, Alcon), or silicone (CeeOn 911, Pharmacia) IOL. All surgery was performed by the same surgeon using a standardized protocol: clear corneal incision, capsulorhexis, phacoemulsification, and in-the-bag IOL implantation. Preoperative and postoperative inflammation was evaluated by measuring aqueous flare preoperatively and 1, 3, 7, 28, 90, and 180 days after surgery using the Kowa FC-1000 laser flare-cell meter. All uveitic eyes were in remission for at least 3 months before surgery., Results: In the uveitic eyes, there was no statistically significant difference in the postoperative course of flare and cell among the 3 IOL groups. Six months after surgery in uveitic eyes, flare values reached preoperative levels and the cell count was lower than preoperatively in all 3 IOL groups. Relative flare values were higher in the eyes with uveitis and a CeeOn 911 IOL; however, the difference between this group and the 2 acrylic IOL groups was not significant., Conclusions: There were no significant differences in inflammation after implantation of foldable IOLs in uveitic eyes. Although absolute flare values and cell counts in eyes with uveitis were higher than in control eyes, primarily because of a damaged blood-aqueous barrier (BAB), BAB recovery was similar between the 2 groups. The changes in the BAB indicate that foldable IOL implantation is safe in uveitic eyes.

Purpose: To determine whether Bcl-2 is expressed on CD4(+ ) lymphocytes in the aqueous humor (AH) and cerebrospinal fluid (CSF) of patients with Vogt-Koyanagi-Harada (VKH) disease, and to determine whether Fas will induce apoptosis of lymphocytes in the CSF., Methods: The percentages of CD4, CD8, CD45RO, Fas, and Bcl-2 positive T lymphocytes in the AH and CSF of eight patients with active VKH and five healthy controls were determined by flow cytometry. Soluble Fas ligand (sFasL) in the CSF was measured by ELISA. Freshly isolated cells from the CSF were cultured with anti-Fas antibody (Ab) and apoptosis was assessed by the TUNEL method., Results: Fas(+) CD4(+) lymphocytes were the predominant lymphocytes in the AH and CSF of VKH patients. Bcl-2 was strongly expressed in these cells. Soluble FasL was also detected in the CSF. The number of apoptotic cells detected by anti-Fas Ab was not significantly increased in the CSF of VKH patients., Conclusions: In spite of the high expression of Fas antigen on CD4(+) cells and the presence of sFasL in the CSF, apoptosis was not observed. Bcl-2 expression may contribute to the regulation of apoptosis of inflammatory cells in the CSF of VKH patients.

Purpose: To evaluate, through differential gene expression of chemokines in iris-ciliary body (I/CB), the extent to which leukocyte recruitment to the ocular anterior segment participates in the pathogenesis of experimental autoimmune uveoretinitis (EAU) in mice., Methods: B10.A mice were immunized with 50 microg of interphotoreceptor retinoid binding protein (IRBP) mixed with complete Freund's adjuvant. Aqueous humor (AqH) was collected at 0, 11, 17, and 28 days and assayed for leukocyte content and protein levels. Enucleated eyes were subjected to histologic analysis. Chemokine gene expression in I/CB was determined at these same time points by a multiprobe ribonuclease protection assay (RPA) system., Results: Inflammation was detected in the anterior chamber (AC) at 11 days and leukocyte recruitment continued thereafter. Polymorphonuclear neutrophils (PMN) were the predominant cell type in the AC, whereas macrophages/monocytes and lymphocytes were predominant in the retina/subretinal space at 17 days. Peak gene expression of macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and MIP-2 and interferon-gamma-inducible protein of 10 kd (IP-10) was detected in I/CB on day 11, whereas peak expression of RANTES and eotaxin was observed at 17 days., Conclusions: Early I/CB peak expression of mRNA for MIP-2 followed by PMN recruitment into the AC, suggests that PMN may play an important role in EAU pathogenesis.

Purpose: We studied the surface expression of intracellular adhesion molecule 1 (ICAM-1) on peripheral and intra-ocular T lymphocytes in patients with active uveitis., Methods: Two-colour flow-cytometric analysis was performed on cells isolated from aqueous humour and peripheral blood of 23 patients with active uveitis and 16 control patients who were to undergo cataract extraction, in order to determine the percentage of cells expressing CD4, CD8 and ICAM-1 (CD54) molecules., Results: In the aqueous humour of patients with uveitis, we found an increase in the percentage of CD4+ and CD8+ lymphocytes, co-expressing the ICAM-1 molecule as compared to control patients (p < 0.0001). In peripheral blood, these uveitis patients exhibited a significant decrease in the percentage of CD4+ICAM-1+ (p = 0.0106) and CD8+ICAM-1+ (p = 0.0014) as compared to control subjects. The comparative study of cells from the aqueous humour and peripheral blood showed that the percentage of CD4+ICAM-1+ cells was higher in the aqueous humour (p < 0.0001). Comparison of the aqueous humour and peripheral blood for CD8+ICAM-1+ cells revealed no significant differences. In addition, we found a high negative correlation for the CD8+ICAM-1+ subset between the aqueous humour and peripheral blood., Conclusions: Our results suggest a greater local participation for CD4+ICAM-1+ cells but not for CD8+ICAM-1+ cells in the pathogenesis of uveitis., (Copyright 2002 S. Karger AG, Basel)

Purpose: To evaluate the role of topical instillation of some antiglaucoma agents on experimental elevation of aqueous flare induced by prostaglandin E(2) (PGE(2)) in pigmented rabbits., Methods: Transcorneal diffusion of PGE(2) (25 microg/mL or 7.09 x 10(-2) mM) with the use of a glass cylinder was achieved to produce aqueous flare elevation in pigmented rabbits. An antiglaucoma agent was topically administered before application of PGE(2). Aqueous flare was measured with a laser flare cell meter., Results: A single instillation of apraclonidine 1.15%, two instillations of epinephrine 1.25%, two instillations of dipivefrin 0.1%, and two instillations and one instillation of dipivefrin 0.04% eye drops inhibited 98%, 96%, 87%, 73%, and 47% of PGE(2)-induced aqueous flare elevation, respectively. Timolol 0.5%, nipradilol 0.25%, dorzolamide 1%, and pilocarpine 2% eye drops had no effects on the increase of PGE(2)-induced flare., Conclusions: Apraclonidine, epinephrine, and dipivefrin eye drops inhibit PGE(2)-induced elevation of aqueous flare in pigmented rabbits.

Objective: To study the effect of tetrandrine (Tet) on the formation of posterior capsular opacification (PCO) after cataract extraction., Methods: Intraocular lens implantation was performed on 48 healthy rabbit eyes, and they were randomly divided into 4 groups: control (A), dexamethasone (B), Tet (C) and Tet with coated IOL (D). Dexamethasone and Tet were postoperatively injected into the subconjunctival space. The number of cells, the contents of protein and malondialdehyde (MDA) in the aqueous were determined at different days postoperatively. The PCO was scored and opaque posterior capsules were weighted., Results: The number of cells, the amount of protein in aqueous, the scales and the wet weight of PCO in the treated eyes were less than that of the control eyes (P < 0.05). In B and C group, except the 14th day content, the postoperative MDA was lower than that in the control group (P < 0.05)., Conclusion: Tet can inhibit the formation of PCO, and has no obvious toxicity to the corneal endothelium.

Purpose: To determine the effect of pyrrolidine dithiocarbamate (PDTC), an antioxidant nuclear factor (NF)-kappaB inhibitor, on the ocular inflammation induced by lipopolysaccharide (LPS)., Methods: Endotoxin-induced uveitis (EIU) was produced by a footpad injection of 200 microg LPS in male Lewis rats. PDTC (200 mg/kg) was injected intraperitoneally 30 minutes before the LPS administration. The number of infiltrating cells and protein concentration in the aqueous humor (AqH) was determined from the AqH collected at 24 hours. Immunohistochemical staining with a monoclonal antibody against activated NF-kappaB was performed to evaluate the effect of PDTC on NF-kappaB activation. Interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)-alpha mRNA expression in the iris-ciliary body (ICB) was determined by RNase protection assay (RPA). The levels of these cytokines and nitric oxide (NO) production were also determined., Results: The number of cells in the AqH was 1100 +/- 254 cells/microL in rats injected with LPS and 90 +/- 43 cells/microL in rats pretreated with PDTC (P < 0.001). The concentration of proteins was significantly lower in the AqH of rats pretreated with PDTC than in those without PDTC. The number of activated NF-kappaB-positive cells in the ICB was reduced by the PDTC treatment. The ICB at 6 hours after LPS injection exhibited increased expression of IL-1beta, IL-6, and TNF-alpha mRNAs, which was decreased after PDTC pretreatment. PDTC also significantly diminished the levels of these cytokines and nitrite-nitrate in the AqH., Conclusions: These results suggest that PDTC reduces ocular inflammation in eyes with EIU by downregulating proinflammatory cytokine expression and by inhibiting the NF-kappaB-dependent signaling pathway.

Purpose: Adrenergic agents decrease intraocular pressure by reducing aqueous humor secretion from ciliary epithelial cells. Since the ionic concentration of aqueous humor contributes to intraocular pressure, we have investigated the effect of (-)-isoproterenol, a beta-adrenergic agonist on the maxi-K( +) channel in rabbit nonpigmented ciliary epithelial (NPE) cells., Methods: Single-channel currents were recorded from the basolateral surface of acutely isolated NPE cells using patch clamp techniques., Results: A calcium dependent maxi-K(+) channel was identified in 31% of cell-attached patches. In the excised condition the channel was activated in presence of calcium. In symmetrical K(+) solution a linear current-voltage relationship and unitary conductance of 158 +/- 15 pS was observed. Replacing K(+) with Na(+) the current-voltage curve shifted to the right and approached a reversal potential for K( +) ( approximately -80 mV). Barium (2 mM) from the intracellular side or iberiotoxin (50 nM) from the extracellular side blocked the channel activity. In cell-attached patches, the beta-receptor agonist (-)-isoproterenol (2.5 microM) increased channel open probability (P(o)) only when applied directly through the patch pipette. beta(2)-adrenoceptor antagonists (ICI-118, 551, l-timolol) blocked the channel activity more efficiently than the beta(1)-adrenoceptor antagonist betaxolol. In excised patches, (-)-isoproterenol increased baseline P(o) 5-fold (0.5 +/- 0.13) when GTP (100 microM) and GTPgammaS (100 microM) were present at the cytosolic surface of the pipette (control; P(o), 0.12 +/- 0.006). GTP augmented baseline channel activity (0.1 +/- 0.004) 7-fold (0.7 +/- 0.03) when (-)-isoproterenol was included in patch pipette., Conclusions: Rabbit NPE cells expressed maxi-K(+) channels on their basolateral surface. The adrenergic agonist (-)-isoproterenol activated these channels via a beta(2)-adrenoceptor that was modulated by a direct G-protein gated pathway.

Purpose: Lymphocytes, monocytes, and macrophages are the predominating immune cells in graft rejection after keratoplasty in animal models. This study focuses on the isolation of immune cells from the anterior chamber of patients with slight, moderate, and severe endothelial immune reactions after penetrating keratoplasty., Methods: Anterior chamber puncture was performed in five patients with cataract without inflammation and without penetrating keratoplasty (C1), in three patients undergoing penetrating keratoplasty without immune reactions (C2), in four patients undergoing penetrating keratoplasty after complete resolution of endothelial immune reactions (C3), in seven patients undergoing penetrating keratoplasty with slight endothelial immune reactions (IMI), in 10 patients undergoing penetrating keratoplasty with moderate endothelial immune reactions (IM2), and in eight patients undergoing penetrating keratoplasty with severe endothelial immune reactions (IM3). In each patient, approximately 0.1 mL of aqueous humor was examined. Cells in suspension were directly centrifuged on glass slides using a Cytospin centrifuge, stained, and evaluated under the light microscope., Results: Groups C1, C2, and C3 did not contain cells. Immune cells were identified in three of seven patients in IM1, in eight of 10 patients in IM2, and in eight of eight patients in IM3. Predominating cells were macrophages and monocytes followed by lymphocytes. Regarding all patients in IMI, IM2, and IM3, a statistically significant correlation between detected cells and patient age, period between penetrating keratoplasty and anterior chamber puncture, or period between first symptoms and anterior chamber puncture could not be revealed. Granulocytes were found statistically significantly less often in patients with high-risk indications, in patients with a history of immune reactions and under immunosuppression. Lymphocytes were found statistically significantly less often in patients with a history of immune reactions., Conclusions: The probability to isolate immune cells from the anterior chamber of patients undergoing penetrating keratoplasty correlates with the severity of the endothelial immune reactions. This study is a first step to evaluate how detailed immunologic findings from animal keratoplasty models fit to clinical reality in patients undergoing keratoplasty. In the next step, cells found in the aqueous humor of patients with endothelial immune reactions should be further characterized directly (determination of molecules on the surface of the cells) or indirectly (determination of cytokine levels in the aqueous humor).

A 60-year-old man visited our clinic with a sudden blurred vision and ocular pain in his right eye occurring 15 years after cataract surgery. The intraocular pressure (IOP) was 55 mmHg in the right eye and gonioscopy revealed a wide open angle with white cortical lens material in the inferior angle. Since the IOP was unable to be controlled with medical therapy, removal of the lens material was performed by irrigation and aspiration. Following surgery, the IOP was decreased to 18 mmHg without medication and the patient's vision recovered to 20/20. The pathology of the aqueous humor showed macrophages with engulfed lens particles.