Intracerebral hemorrhage due to vasculitis following COVID-19 vaccination: a case report.

Several vaccines have been approved worldwide for the prevention of morbidity and mortality against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the development of these vaccines has raised concerns regarding their adverse effects. Herein, we report the first case of intracerebral hemorrhage (ICH) due to vasculitis after the first dose of mRNA vaccine (BNT162b2, Pfizer/BioN-Tech). Although this case cannot demonstrate a direct relationship between COVID-19 vaccination and vasculitis, the clinical and histological features of this patient are highly consistent with the adverse effects of COVID-19 vaccine.

[Factors Affecting Uncorrected Visual Acuity following Implantation of Apodized Diffractive Intraocular Lenses].

PURPOSE: To investigate the preoperative factors affecting postoperative uncorrected visual acuity (UCVA) equal to or more than 20/20. SUBJECTS AND METHODS: One hundred sixty seven eyes receiving apodized diffractive multifocal intraocular lenses (SN6AD1) were included in this study. In the eyes with corneal astigmatism of less than 1.0 D, a 2.4 mm clear temporal corneal incision was created. In those equal to or more than 1.0 D, 4.1 mm steepest meridian clear corneal incisions and/or limbal relaxing incisions were conducted. Preoperative factors affecting postoperative UCVA equal to or more than 20/20 were assessed by univariate analysis and multiple logistic regression analysis. RESULTS: Patient's age and preoperative corneal astigmatism were significant in both univariate analysis and multiple logistic regression analysis. The ratio of postoperative UCVA equal to or more than 20/20 was significantly lower in the patients 70 years or older and in the eyes with corneal astigmatism equal to or more than 1.5 D. CONCLUSION: Multifocal toric intraocular lenses provide better surgical results in eyes with corneal astigmatism, however patient's age needs to be considered as a factor affecting UCVA.

Differential effects of myocilin and optineurin, two glaucoma genes, on neurite outgrowth.

Myocilin and optineurin are two genes linked to glaucoma, a major blinding disease characterized by progressive loss of retinal ganglion cells (RGCs) and their axons. To investigate the effects of force-expressed wild-type and mutant myocilin and optineurin on neurite outgrowth in neuronal cells, we transiently transfected cells with pEGFP-N1 (mock control) as well as myocilin and optineurin plasmids including pMYOC(WT)-EGFP, pMYOC(P370L)-EGFP, pMYOC(1-367)-EGFP, pOPTN(WT)-EGFP, and pOPTN(E50K)-EGFP. PC12 cells transfected with pEGFP-N1 produced, as anticipated, long and extensive neuritis on nerve growth factor induction. The neurite length in those cells transfected with myocilin constructs was shortened and the number of neurites was also reduced. A similar inhibitory effect on neurite outgrowth was also elicited by myocilin transfection in RGC5 cells. In contrast, neither transfection of the optineurin constructs pOPTN(WT)-EGFP and pOPTN(E50K)-EGFP nor the myocilin and optineurin small-interfering RNA treatments induced significant alterations in neurite outgrowth. Transfection with the wild-type optineurin construct, but not with that of the wild-type myocilin, increased the apoptotic activity in cells. These results demonstrated that the two glaucoma genes, myocilin and optineurin, exhibited differential effects on neurite outgrowth. They may contribute to the development of neurodegenerative glaucoma via distinct mechanisms.

Suppression of choroidal neovascularization in lectin-like oxidized low-density lipoprotein receptor type 1-deficient mice.

PURPOSE: To elucidate the role of the scavenger receptor, lectin-like oxidized low-density lipoprotein receptor type 1 (LOX-1), in the formation of choroidal neovascularization (CNV). METHODS: CNV was induced by laser photocoagulation of the ocular fundus in mice. The expression of LOX-1 mRNA and protein after laser injury was determined by real-time RT-PCR and Western blot analysis. Gelatin zymography was used to measure the activity of matrix metalloproteinase (MMP)-2 and pro-MMP-9, and ELISA was used to determine monocyte chemoattractant protein (MCP)-1 and vascular endothelial growth factor (VEGF) levels. At 14 days after laser injury, the extent of CNV was evaluated by fluorescein angiography and lectin staining using confocal microscopy. RESULTS: In wild-type mice, the relative expression level of LOX-1 mRNA compared with the control increased significantly 6 hours after laser injury and peaked 12 hours after laser injury (P = 0.011 and P = 0.0006, respectively), and the expression of LOX-1 protein was also detected 1 and 3 days after laser injury. Increases in MMP-2, pro-MMP2, and pro-MMP-9 after laser injury were reduced in LOX-1-deficient mice compared with wild-type mice. At 3 days after laser injury, increases in MCP-1 and VEGF significantly decreased in LOX-1-deficient mice compared with wild-type mice (P = 0.014 and P = 0.001, respectively). Morphometric analyses revealed that the induction of CNV formation was significantly inhibited in LOX-1-deficient mice. CONCLUSIONS: These results suggest that LOX-1 plays an important role in the formation of CNV. This scavenging system might thus be a novel therapeutic target for CNV.

Downregulation of LIM kinase 1 suppresses ocular inflammation and fibrosis.

PURPOSE: The purpose of this study was to determine if downregulation of LIM kinase 1 (LIMK1) by genetic deletion or direct application of LIMK1-targeted siRNA could suppress TGF-beta mediated ocular inflammation and fibrosis. METHODS: LIMK1 specific siRNAs designed from the human sequence were transfected into human corneal fibroblasts in culture. Immunofluorescence and immunoblotting were performed to examine the fibronectin assembly. The effects of LIMK1 downregulation on actin cytoskeleton organization and focal adhesion formation were studied. A wound closure assay was used to assess cell migration in in vitro fibroblast cultures. The in vivo effects of LIMK1 genetic deletion or downregulation by mouse siRNA were evaluated in a mouse model of ocular inflammation generated by subconjunctival injection of phosphate buffered saline and latex beads. Cellularity on tissue sections was examined after staining with hematoxylin and eosin. Anti-CD45 antibody was used for the leukocyte detection. RESULTS: Downregulation of LIMK1 in cultured corneal fibroblasts impaired fibronectin secretion and assembly, diminished actin polymerization and focal adhesion formation, and retarded cell migration. In the mouse model of ocular inflammation, both genetic deletion and downregulation of LIMK1 by siRNA significantly reduced inflammatory response. CONCLUSIONS: Downregulation of LIMK1 was efficacious to decrease the ocular inflammation. We disclose a possibility that LIMK1 may mediate TGF-beta-dependent signaling during ocular inflammation. A direct application of siRNA into eyes to downregulate LIMK1 expression may provide a novel therapy for suppression and prevention of ocular inflammation and fibrosis.

Long-term relationship between intraocular pressure and visual field loss in primary open-angle glaucoma.

PURPOSE: To investigate the dependence upon intraocular pressure (IOP) of the progression of visual field defects in eyes with primary open-angle glaucoma (POAG), in which the mean IOP was maintained at < or =21 mm Hg. METHODS: This study involved 100 eyes with POAG, which were followed up for > or =5 years. The mean IOP levels were maintained at < or =21 mm Hg during the follow-up period. The relationship between the IOP and the progression of visual field defects, which was scored using the Advanced Glaucoma Intervention Study criteria, was investigated retrospectively. RESULTS: Compared with the baseline scores, the visual field defect scores had significantly worsened by the end of the follow-up period (P<0.0001, Wilcoxon paired signed rank test). The change in the visual field defect score (2.5+/-0.5) in eyes with average IOP levels of > or =16 mm Hg (n=36) was significantly greater (P=0.031, Mann-Whitney U test) than the change (1.3+/-0.3) in eyes with average IOP levels of <16 mm Hg (n=64). Moreover, IOP of > or =18 mm Hg made a major contribution to the aggravation of visual field defects in eyes with POAG. CONCLUSIONS: Eyes with POAG and with mean IOP levels maintained at < or =21 mm Hg underwent IOP-dependent progression of their visual field defects. Our results suggest that further IOP lowering would be beneficial in such cases.

Rho GTPase and cAMP/protein kinase A signaling mediates myocilin-induced alterations in cultured human trabecular meshwork cells.

Myocilin is a gene linked to the most common form of glaucoma, a major blinding disease. The trabecular meshwork (TM), a specialized eye tissue, is believed to be involved, at least in part, in the development of glaucoma. The myocilin expression is known to be up-regulated by glucocorticoids in TM cells, and an altered myocilin level may be the culprit in conditions such as corticosteroid glaucoma. Wild type myocilin, when transfected into cultured human TM cells, induced a dramatic loss of actin stress fibers and focal adhesions. Myocilin transfectants displayed a heightened sensitivity to trypsin. Adhesion to fibronectin, collagens, and vitronectin was compromised. The fibronectin deposition and the levels of fibronectin protein and mRNA were also reduced in myocilin transfectants. The fibronectin deposition could be restored by treatment with lysophosphatidic acid, a Rho stimulator. Assays further revealed that upon myocilin overexpression, the activity of RhoA was diminished, whereas the cAMP level and the protein kinase A (PKA) activity were augmented. Myocilin protein did not affect actin polymerization. The collapse of actin stress fibers and increased trypsin sensitivity from myocilin transfection could be reverted by co-expression of constitutively active RhoA or by treatment with PKA inhibitor H-89. The PKA activity, however, was not modified by co-expression of either constitutively active or dominant negative RhoA. These results demonstrate that myocilin has a de-adhesive activity and triggers signaling events. cAMP/PKA activation and the downstream Rho inhibition are possible mechanisms by which myocilin in overabundance may lead to TM cell or tissue damage.

Mitochondrial association of myocilin, product of a glaucoma gene, in human trabecular meshwork cells.

The trabecular meshwork (TM), an ocular tissue next to the cornea, is a major site for regulation of the aqueous humor outflow. Malfunctioning of this tissue is believed to be responsible for development of glaucoma, a major blinding disease. Myocilin is a gene directly linked to the most common form of glaucoma. Its protein product has been localized to both intra- and extra-cellular sites in TM cells. This study was to investigate the association of myocilin with mitochondria in TM cells. In vitro mitochondrial import assays showed that myocilin was imported to the TM mitochondria, targeting to mitochondrial membranes and/or the intermembrane space. The targeting was mediated mostly via the amino-terminal region of myocilin. When myocilin expression was induced either by treatment with dexamethasone or transfection with a myocilin construct, the mitochondrial membrane potential in TM cells, as assessed by JC-1 staining, was lowered. Subcellular fractionation and Western blot analyses confirmed that a portion of myocilin sedimented with the mitochondrial fractions. Upon anti-Fas treatment to provoke apoptosis, an increase of myocilin distribution in cytosolic fraction was observed, suggesting that myocilin was partially released from mitochondrial compartments. These results confirmed the association of myocilin with TM cell mitochondria and indicated that myocilin may have a proapoptotic role in TM cells.

Frequency and risk factors for intraocular pressure elevation after posterior sub-Tenon capsule triamcinolone acetonide injection.

PURPOSE: This study investigated the effects of posterior sub-Tenon capsule (PST) injection of triamcinolone acetonide (TA) on intraocular pressure (IOP) in the human eye. METHODS: The study included 115 patients who received PST injections of 40-mg TA to treat macular edema with diabetic retinopathy (n=57), branch retinal vein occlusion (n=35), central retinal vein occlusion (n=13), or other disorders (n=10). IOP measurements were performed on the day of injection, and 0.5, 1, 2, 3, 6, 9, and 12 months later. RESULTS: In 26 (22.6%) of the 115 eyes, an IOP of 24 mm Hg or higher was observed during the 12-month follow-up period after PST TA injection. IOP elevation significantly correlated with young age, but not with past history of diabetes mellitus or systemic hypertension, sex, or type of retinal disease with macular edema. In total, 23 eyes were treated with antiglaucoma medications to control elevated IOP (24 mm Hg or higher). External trabeculotomy was performed in 1 case where medications failed to correct elevated IOP. CONCLUSIONS: PST TA injection is associated with high rates of steroid-induced IOP elevation in eyes with previously normal IOP. However, IOP elevation may be less common after PST injection than after intravitreal injection. Our findings indicate that IOP must be carefully monitored after PST TA injection.

Stability of the central visual field after modern trabeculectomy techniques in eyes with advanced glaucoma.

PURPOSE: To review the surgical results and complications of trabeculectomy techniques in patients with advanced glaucoma and threatened fixation. METHODS: Trabeculectomy had been carried out on 49 advanced glaucoma patients (49 eyes) using mitomycin C and postoperative laser suture lysis. The clinical records prior to and 2 months after surgery were reviewed, and the long-term surgical outcomes were determined. RESULTS: Two months after surgery there were no eyes with fixation loss. Intraocular pressure (IOP) levels were reduced from 22.8 +/- 6.0 to 11.7 +/- 4.7 mmHg. Kaplan-Meier survival analysis showed that the success rate in achieving IOPs of 15 mmHg or lower 5 years after surgery was 70%. The chance of visual acuity remaining within two lines of the preoperative level was 75%. In 29 of the 49 eyes, visual acuities remained at their preoperative level at the time of the final visit, but had decreased to less than 0.1 in three eyes (cataract progression, n = 2; fixation loss, n = 1). CONCLUSION: The results suggest that laser suture lysis and stepwise management of IOP levels, which are performed as part of the modern postoperative management of trabeculectomy, decrease the frequency of fixation loss during the early postsurgical phase.

Rho-associated protein kinase inhibitor, Y-27632, induces alterations in adhesion, contraction and motility in cultured human trabecular meshwork cells.

We investigated the roles of Rho-associated protein kinase (ROCK) in regulating activities such as adhesion, contraction and migration in cultured human trabecular meshwork (TM) cells. Human TM cells in culture were treated with Y-27632, a specific ROCK inhibitor. Trypan blue exclusion test and TUNEL staining showed little or no direct toxicity of Y-27632 on TM cells. By MTT assay, Y-27632 did not significantly affect the proliferation of TM cells. The cell adhesion assay showed that Y-27632 promoted the cell adhesiveness to both fibronectin and collagen type I in a dose-dependent manner. Collagen gel contraction activity of TM cells was significantly inhibited by the treatment of Y-27632 in a dose-dependent manner. The addition of Y-27632 accelerated motility of TM cells in wound healing assay. Phosphorylated LIM kinase 2 and cofilin, related to actin bundling and integrin clustering, were dephosphorylated (activated) by Y-27632. In conclusion, Y-27632 elicits profound effects on TM cell activities including adhesion, gel contraction, and cell motility. These Y-27632-induced changes of TM cells may be relevance to the physiology of the aqueous outflow system.

NMDA-induced retinal injury is mediated by an endoplasmic reticulum stress-related protein, CHOP/GADD153.

We investigated the role of an endoplasmic reticulum stress-associated protein, CHOP/GADD153, after NMDA-induced mouse retinal damage. After injection of NMDA into the vitreous, TUNEL-positive cells were detected in the retinal ganglion cell layer (GCL) and inner nuclear layer (INL) at 6 h after NMDA injection, and these gradually increased in number up to 24 h. Analysis by real-time RT-PCR revealed that CHOP mRNA was induced by about 3-fold, at 2 h after NMDA injection. Immunoreactivity for the CHOP protein was intense in cells of the GCL following NMDA treatment. Immunoblot analysis showed that NMDA injection increased the expression of CHOP protein in the retina. Compared with wild-type mice, CHOP/ mice were more resistant to NMDA-induced retinal cell death as determined by TUNEL assay. At 7 days after NMDA treatment, the thickness of the inner plexiform layer and INL were larger in CHOP/ mice than in wild-type mice. The number of residual cells in the GCL following NMDA treatment was significantly higher in CHOP/ mice than in wild-type mice. In conclusion, CHOP is induced in mouse retina by NMDA treatment, and CHOP/ mice are more resistant to NMDA-induced retinal damage, suggesting that CHOP plays an important role in NMDA-induced retinal cell death.

Expression of a chondroitin sulfate proteoglycan, versican (PG-M), during development of rat cornea.

PURPOSE: To understand the role of chondroitin sulfate proteoglycans during the development of rat cornea, expression of chondroitin sulfate and versican (PG-M) was studied. METHODS: Chondroitin sulfate and keratan sulfate in rat cornea were analyzed by immunohistochemical techniques. Reverse transcription polymerase chain reaction (RT-PCR) for chondroitin sulfate proteoglycans was performed. Versican expression was studied by RT-PCR, immunohistochemical, and dot blot analyses. Expression of hyaluronan was evaluated histochemically using biotinylated hyaluronan binding protein. RESULTS: Chondroitin sulfate was abundant in rat cornea at postnatal day 1 (P1) and became undetectable at P14. RT-PCR analysis showed that versican mRNA was highly expressed at P1 but was little expressed at P42. mRNAs for other chondroitin sulfate proteoglycans including biglycan, aggrecan, and decorin did not change much between P1 and P42. Expression for all versican splicing isoforms (V0-V3) was detectable from P1 through P14 but was undetectable after P21. mRNA for V0, the largest form with many chondroitin sulfate binding sites, decreased markedly in early stages from P1 to P14, whereas mRNA for V3, the shortest form with no chondroitin sulfate binding site, increased. mRNAs for middle-sized forms, V1 and V2, remained little changed during these periods. Immunohistochemical and dot blot analyses showed that versican is highly expressed at early stages of development and little expressed at adulthood. Similarly, hyaluronan, a versican-bound glycosaminoglycan, was highly expressed at early stages and little expressed at adulthood. CONCLUSIONS: Versican and hyaluronan, which can form a large molecular complex, may play an important role in the early phase of corneal development.

Genetic polymorphisms in the angiotensin II receptor gene and their association with open-angle glaucoma in a Japanese population.

PURPOSE: The local renin-angiotensin system (RAS) is present in the ciliary body and plays a role in regulating aqueous humor dynamics and thus intraocular pressure (IOP). The purpose of this study was to determine whether gene polymorphisms in the RAS increase the risk of development of glaucoma in the Japanese. METHODS: A case-control study was performed in 698 Japanese subjects: 190 patients with primary open-angle glaucoma (POAG), 268 patients with normal-tension glaucoma (NTG), and 240 normal subjects. Ten polymorphisms in seven genes-AGT/Thr174Met and AGT/Met235Thr; REN/I8-83G-->A; ACE/insertion(I)-deletion(D); CMA/-1930A-->G; AGTR1/-731T-->G, AGTR1/-521C-->T, and AGTR1/1166A-->C; AGTR2/3123C-->A; and CYP11B2/-344T-->C were examined. The age, IOP, and visual field defects, all at diagnosis, were examined to determine whether they were associated with the polymorphisms. The effects of oral angiotensin II receptor blocker (ARB) on IOP were examined in association with the AGTR1 and AGTR2 polymorphisms in 20 normal subjects. RESULTS: Of the 10 polymorphisms, the AGTR2/3123C-->A polymorphisms had a significantly different distribution in female patients with NTG; the frequency of the CA+AA genotypes was significantly higher than in female control subjects (P = 0.0095 for CC versus CA+AA). Although no significant difference was seen in the clinical characteristics of female patients with NTG who carried the AGTR2/3123C-->A genotype, patients with CC in the AGTR2 gene had significantly worse visual field scores if they carried ACE/ID+DD (i.e., D carriers; P = 0.012). ARB significantly lowered IOP in normal subjects, but the male subjects with the AGTR2/3123A genotype had significantly less lowering of IOP than those with the C genotype (P = 0.014). CONCLUSIONS: Angiotensin II receptor gene polymorphisms may be associated with the risk of glaucoma in the Japanese population.

A case of Chandler's syndrome revealed by ultrastructural studies of the trabecular meshwork.

PURPOSE: To investigate ultrastructural changes in the aqueous outflow route and discuss the mechanisms associated with intraocular pressure (IOP) elevation in a patient with presumably early stage Chandler's syndrome. METHODS: A 47-year-old man underwent trabeculectomy because of elevated IOP. A specimen obtained during surgery was studied by transmission electron microscopy. RESULTS: Electron microscopy showed the presence of a monolayer composed of corneal endothelium-like cells and thick basement membrane-like material. Neovascularization was also observed in the corneoscleral trabeculum. CONCLUSIONS: Our results indicate that several mechanisms, including the formation of basement membrane-like tissue, infiltration of inflammatory cells and neovascularization, might contribute to the elevation of IOP in Chandler's syndrome. These may occur even when there is no history of conspicuous inflammatory reaction in the anterior ocular segments.

Morphological damage in rabbit retina caused by subretinal injection of indocyanine green.

PURPOSE: To investigate the effects of subretinal indocyanine green (ICG) on retinal morphology in rabbit eyes. METHODS: Retinal bleb detachments were produced by injections of ICG at dosages of 25 mg/ml, 5 mg/ml, and 0.5 mg/ml or with an injection of balanced salt solution (BSS) into the subretinal space of albino rabbit eyes. Morphological change was assessed by light and transmission electron microscopy from the viewpoint of dose and time. Some sections were also probed with the TUNEL technique to detect apoptotic cells. RESULTS: At 14 days after subretinal injection of BSS and 0.5 mg/ml ICG, the structure of the retina was well preserved. However, injections of 5 mg/ml or 25 mg/ml caused thinning of the retina, especially loss of the outer retinal layer. In eyes injected with 5 mg/ml ICG, the photoreceptors began disappearing within 3 days after the injection and over time showed the development of retinal atrophy. TUNEL-positive cells appeared abundantly in the photoreceptor layers 1 and 3 days after the injection of 5 mg/ml ICG. Transmission electron microscopy confirmed apoptosis in the photoreceptors. CONCLUSION: These data indicate that subretinal ICG induces apparent morphological damage of the retina in a dose-dependent manner.

Expression of glycosaminoglycans during development of the rat retina.

PURPOSE: To investigate the spatiotemporal expression of glycosaminoglycans during development of the rat retina. METHODS: Hyaluronan and sulfated glycosaminoglycans, including chondroitin sulfate, heparan sulfate and keratan sulfate were detected using biotinylated hyaluronan binding protein, immunohistochemical analysis, respectively, in the rat retina at various stages of development. RESULTS: Hyaluronan was expressed in the nerve fiber layer, inner plexiform layer and outer plexiform layer during early postnatal stages (postnatal day 1-14; P1-P14) and was undetectable after P21. In contrast, hyaluronan was faintly observed in the photoreceptor layer on P7, and gradually increased up to P49. The spatiotemporal expression pattern of chondroitin sulfate was similar to that of hyaluronan. Heparan sulfate was also detected in the nerve fiber layer, inner plexiform layer and outer plexiform layer during early postnatal stages (P1-P14). In addition, heparan sulfate was expressed in the inner limiting membrane during all stages of development. Keratan sulfate was not detected in the retina at any stage of development. CONCLUSIONS: Hyaluronan, chondroitin sulfate and heparan sulfate are expressed in nerve fiber-rich layers during early postnatal stages and may regulate neurite outgrowth. In adulthood, both hyaluronan and chondroitin sulfate are expressed in the photoreceptor layer and may consist of the interphotoreceptor matrix. In addition, heparan sulfate is expressed in the inner limiting membrane throughout the various stages of development and may be associated with the structure of the inner limiting membrane.

Lens epithelium-derived growth factor: neuroprotection on rat retinal damage induced by N-methyl-D-aspartate.

The purpose of this study is to investigate possible neuroprotective effects of lens epithelium-derived growth factor (LEDGF) against cell death induced by N-methyl-D-aspartate (NMDA) in the rat retina. LEDGF and/or NMDA were intravitreally injected into rat eyes. NMDA-induced retinal death and protective effects of LEDGF were evaluated by morphometric analysis, cell numbers in the ganglion cell layer (GCL) and the thickness of the inner plexiform layer (IPL). Retrograde labeling with a fluorescent tracer (Fluoro-Gold) was applied for counting retinal ganglion cells (RGCs) that survived after NMDA injection. Terminal deoxyribonucleotidyl transferase (TdT)-mediated fluroscein-16-dUTP nick end-labeling (TUNEL) staining was used to evaluate of retinal cell death. Morphometric analysis and retrograde labeling analysis showed that retinal damage induced by NMDA was protected significantly by LEDGF. TUNEL assay revealed that pretreatment with LEDGF prevents NMDA-induced apoptosis. Retinal damage (ganglion and amacrine cells) induced by NMDA was protected by an intravitreal injection of LEDGF.

Secondary glaucoma in patients with familial amyloidotic polyneuropathy.

OBJECTIVE: To elucidate the clinical features and surgical outcomes of the treatment of secondary glaucoma associated with transthyretin (TTR)-related familial amyloidotic polyneuropathy (FAP). DESIGN: Retrospective case study. PARTICIPANTS: Forty-nine Japanese patients with FAP. METHODS: For all patients, measurement of best-corrected visual acuity, intraocular pressure, and visual fields as well as slitlamp and ocular fundus examinations were conducted and compared. In addition, the exact mutation of the amyloidogenic TTR variants was analyzed for all 49 patients with FAP. The TTR mutations included amyloidogenic TTR (ATTR) Val30Met in 41 patients, ATTR Tyr114Cys in 6, ATTR Ser50Ile in 1, and a compound heterozygous mutation of ATTR Val30Met + Arg104His in 1. RESULTS: The onset of secondary glaucoma was defined as elevation of intraocular pressure and glaucomatous changes in visual field defects. Secondary glaucoma was detected in 12 (24%) of the 49 patients. The incidence of secondary glaucoma in patients with the Val30Met mutation (17%) was lower than for the other FAP genotypes (P =.02 using the chi(2) test). Of 20 glaucomatous eyes, amyloid deposition on the pupil and anterior surface of the lens was found in 18 eyes. Amyloid deposition was found prior to glaucoma in 11 eyes and at the first visit to our clinic in another 7 eyes. In the 11 eyes in which the onset of glaucoma occurred following amyloid deposition along the pupil, the mean +/- SD period between the onsets of pupillary amyloid deposition and glaucoma was 2.55 +/- 1.43 years (range, 0.2-4.0 years). Further statistical analyses revealed significant relationships between the onset of secondary glaucoma and both amyloid deposition (P<.001) and vitreous opacity (P<.001). Surgical treatment was required in 15 (75%) of the 20 glaucomatous eyes. In 9 (81%) of the 11 eyes that underwent trabeculectomy, the intraocular pressure was well controlled at or lower than 20 mm Hg during the follow-up period. In the eyes that underwent combined trabeculotomy and sinusotomy (2 eyes), nonpenetrating trabeculectomy (1 eye), or a cyclodestructive procedure (1 eye), the intraocular pressure was poorly controlled. CONCLUSIONS: Glaucoma is not a rare condition in patients with FAP, especially because liver transplantation now enables patients with FAP to live longer. Careful observation of amyloid deposition along the pupil allows the prediction of glaucoma onset.

Induction of citrulline-nitric oxide (NO) cycle enzymes and NO production in immunostimulated rat RPE-J cells.

Nitric oxide (NO) has been implicated in many physiological and pathological conditions in the eyes. The induction of inducible NO synthase (iNOS) and NO production have been noted in immunostimulated retinal pigment epithelial (RPE) cells. Cellular NO production depends on the availability of arginine, a substrate for NOS. Arginine can be regenerated from citrulline, another product of the NOS reaction, by argininosuccinate synthetase and argininosuccinate lyase, forming the citrulline-NO cycle. When rat RPE-J cells were treated with interferon-gamma (IFNgamma), tumor necrosis factor-alpha (TNFalpha) and lipopolysaccharide (LPS), and expression of the citrulline-NO cycle enzymes and related enzymes was analyzed, iNOS and argininosuccinate synthetase were highly induced at both mRNA and protein levels. On the other hand, argininosuccinate lyase was not induced. Among other related enzymes and transporters, mRNA for cationic amino acid transporter (CAT)-1 was weakly induced, whereas those for CAT-2, arginase I and II, ornithine aminotransferase and ornithine decarboxylase remained little changed. NO was produced by cells after stimulation with TNFalpha, IFNgamma and LPS. The induction of iNOS mRNA and the production of NO by these immunostimulated cells was further enhanced by cAMP. NO was produced from citrulline as well as from arginine. Our findings indicate that in activated RPE-J cells citrulline-arginine recycling is important for NO production.