The expression of matrix metalloproteinases and their inhibitors in corneal fibroblasts by alarmins from necrotic corneal epithelial cells

Purpose

Sterile ulceration is frequently observed in the cornea following persistent corneal epithelial damage. We examined the effect of alarmins released by necrotic corneal epithelial cells (HCE) on the production of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) by corneal fibroblasts.

Methods

IL-1α and high-mobility group box 1 protein (HMGB1) released into the supernatant derived from necrotic HCE cells were measured with enzyme-linked immunosorbent assay (ELISA). MMPs and TIMPs produced by corneal fibroblasts, stimulated with the supernatant from necrotic HCE cells, were analyzed and measured with protein array and ELISA. To investigate dynamic expression of alarmins in the corneal epithelium, we used immunohistochemistry to observe the expression of human IL-1α in the corneal epithelium of human IL-1α Tg mice with or without cryopexy. We also investigated the expression of MMPs in corneal stroma of the mice treated with cryopexy, using RT-PCR.

Results

We detected IL-1α and HMGB-1 in the supernatant of necrotic HCE cells. These supernatants increased the expression of MMP-3 and MMP-1, and decreased that of TIMP-1 and TIMP-2 in human corneal fibroblasts. Almost always these were inhibited by IL-1 receptor antagonist. Recombinant IL-1α increased the production MMP-3 and MMP-1 in corneal fibroblasts. After cryopexy of the epithelium of human IL-1α Tg mice, the expression of human IL-1α was recognized in the cytoplasm but not nucleus of epithelial cells. The level of MMP-3 and MMP-1 mRNAs was elevated in the corneal stroma in mice treated with cryopexy.

Conclusion

Alarmins, especially IL-1α, released from necrotic HCE cells may play an important role in the expression of MMPs and TIMPs by corneal fibroblast, resulting in sterile ulceration.

     

Infection of endotheliotropic human cytomegalovirus of trabecular meshwork cells

Purpose

Human cytomegalovirus (HCMV) infections can cause endotheliitis which is associated with an elevation in the intraocular pressure (IOP). However, the mechanism of the IOP elevation has not been established. The purpose of this study was to determine whether HCMV strains which are capable of infecting corneal endothelial cells can also replicate and induce anti-viral responses, and can reorganize the actin cytoskeleton in trabecular meshwork cells.

Study design

Experimental study design.

Methods

Cultured primary human trabecular meshwork cells (HTMCs) were infected with the Towne or TB40/E strains of HCMV. TB40/E is trophic for vascular endothelial and corneal endothelial cells. Real-time PCR, western blot, and fluorescent immunostaining have been used to determine whether HCMV-infected HTMCs will support the expression of viral mRNA and protein, allow viral replication, and elicit anti-viral host responses. We also determined whether lytic replication was present after HCMV infection.

Results

HCMV infection led to the expression of viral mRNA and proteins of IE1, glycoprotein B(gB), and pp65. TB40/E infection induced interferon-β, a sign of host anti-viral immune response and monocyte chemotactic protein-1 (MCP-1) as IOP-related chemokine. Together with the induction of the regulators of actin cytoskeleton, myosin phosphatase Rho interacting protein (MPRIP) and MCP-1, TB40/E induced a high level of expression of viral proteins, including IE1, gB, and pp65 as well as actin stress fiber formation, and achieved pathogenically high viral titers.

Conclusions

Human trabecular meshwork cells support the replication of endotheliotropic TB40/E strain of HCMV which indicates that this strain may have high virulence for trabecular meshwork.

     

Acute visual loss and intraretinal hemorrhages associated to energy drink consumption

Purpose

To report the association of acute visual loss secondary to intraretinal hemorrhages and energy drink consumption.

Methods

Case report and literature review.

Results

A 48-year-old hypertensive man developed an elevation in systemic blood pressure, tachycardia, and acute visual loss secondary to intraretinal hemorrhages shortly after drinking several cans of energy drinks.

Conclusion

High consumption of energy drinks may lead to intraretinal hemorrhages and acute visual loss.

     

Serum and aqueous humor concentrations of interleukin-27 in diabetic retinopathy patients

Purpose

Interleukin (IL)-27 has been reported to possess anti- and proinflammatory properties in several immune related-disorders, but its role in diabetic retinopathy is still elusive. Here, we aimed to (i) evaluate IL-27 concentrations in serum and aqueous humor of diabetic patients with or without retinopathy and (ii) test whether IL-27 is correlated with some risk factors of diabetic retinopathy.

Methods

The study comprised 60 diabetic patients with and without retinopathy along with 20 healthy controls. Serum and aqueous humor concentrations of IL-27 were assessed by ELISA.

Results

The mean of IL-27 concentration in aqueous humor in patients with diabetic retinopathy (6.7 ± 2.7 ng/L) was significantly elevated in comparison with either diabetic patients without retinopathy (4.6 ± 0.5 ng/L) or healthy control group (4.1 ± 0.8 ng/L). Besides, IL-27 concentration in aqueous humor was positively correlated with serum glucose, lipid profile and glycated hemoglobin (HbA1c).

Conclusions

Based on this study, IL-27 is implicated in the pathogenesis of diabetic retinopathy and positively correlates with the disorder progression.

     

Plasma and whole-blood chemokine levels in patients with Behcet's disease

Background

Chemokines are a family of chemoattractants of leukocytes that play a critical role for leukocyte recruitment in various inflammatory diseases. The purpose of this study is to investigate the involvement of chemokines, interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) in the peripheral blood, with a special reference to disease activities of the patients with Behçet's disease (BD).

Methods

The study population consisted of totally 55 patients with BD who had panuveitis (20 patients with active BD, 35 patients with inactive BD) as well as 19 healthy volunteers as control. Disease activity was defined according to the existence of ocular inflammation. IL-8 and MCP-1 concentration levels in the plasma and whole-blood samples were measured by enzyme-linked immunosorbent assay. Whole-blood samples were obtained by lysing cell membranes of peripheral blood cells.

Results

Most of the plasma IL-8 samples were below the detectable limit. Whole-blood IL-8 levels were readily measured. The levels in the patients with active BD were significantly higher than the other two groups. The patients with active and inactive BD showed higher plasma and whole-blood levels of MCP-1 than controls. The plasma and whole-blood MCP-1 levels of the samples collected at the same time showed a linear correlation.

Conclusion

A close relationship was found to exist between the cell-associated IL-8 and the disease activity, while a persistent role of MCP-1 was observed in BD. Measuring the whole-blood levels of chemokines is useful for monitoring the disease activity.

     

Non-arteritic anterior ischemic optic neuropathy (NA-AION) after intravitreal injection of bevacizumab (Avastin®) for treatment of angoid streaks in pseudoxanthoma elasticum

Background

To report a case of nonarteritic anterior ischemic optic neuropathy (NA-AION) following intravitreal injection of bevacizumab (Avastin®).

Methods

Interventional case report with an 18-month follow-up.

Results

A 51-year-old male with pseudoxanthoma elasticum presented with NA-AION 2 weeks after treatment with intravitreal of bevazicumab (Avastin®) for choroidal neovascularisation secondary to angioid streaks. Except from a small optic disc without cupping he did not show further risk factors.

Discussion

Risk of NA-AION should be taken into consideration when deciding for intravitreal application of drugs including anti-vascular endothelial growth factors (VEGF) agents like bevacizumab (Avastin®) in the treatment of retinal vascular diseases.

     

Transplantation of rat embryonic stem cell-derived retinal cells restores visual function in the Royal College of Surgeons rats

Aim of study

To evaluate the feasibility of transplantation of embryonic stem cell (ESC)-derived retinal cells in the treatment of retinal degeneration.

Materials and methods

Rat ESCs were isolated and induced into retinal progenitor cells (RPCs) in vitro, which were subsequently induced into retinal pigment epithelium cells (RPEs) and photoreceptors (PRCs). All cells were identified by Western blot detection of their specific markers. RPEs and PRCs were, respectively, injected into the retina of Royal College of Surgeons (RCSs) rats. Control group was injected with PBS. Post-transplantation visual function was determined by electroretinography (ERG). The histology of the whole eye was compared by H&E staining.

Results

RPEs and PRCs were successfully derived from rat ESCs through the two-step differentiation as indicated by the presence of ESC- (Oct-3/4, Nanog, TRA-1-60 and TRA-1-81), RPC- (Rx, Mitf, Pax6 and Chx10), RPE- (RPE65 and keratin) and PRC-specific markers (blue opsin, red/green opsin, recoverin and rhodopsin) in Western blot. The amplitude of ERG a- and b-wave in RPE- and PRC-transplanted groups at week 2 and 10 after transplantation was markedly higher compared with PBS controls. Retinal injury and vascular stress response was not detected in any of the RCS rats after transplantation.

Conclusion

The developed stepwise protocol can derive retinal cells from ESCs. Transplantation of these retinal cells can restore visual function of RCS rats. Our study provides evidence for potential clinical application of ESC-based cell therapy for retinal degeneration.

     

Upper fornix approach combined with a superior lateral cantholysis: a minimally invasive approach to the superonasal intraconal space

Purpose

To present a new minimally invasive approach to the deep superonasal orbit.

Methods

This retrospective study reviewed seven consecutive patients who underwent orbital surgery using an upper conjunctival fornix approach combined with a superior lateral cantholysis for tumors in the superonasal intraconal space. Charts were reviewed for demographic, radiological, clinical, and surgical data including surgical outcome and morbidities for each patient.

Results

Six benign tumors of the superonasal intraconal orbit were successfully exposed and removed using this approach, and one malignant tumor was biopsied for diagnosis. Histopathology revealed cavernous haemangioma (3 cases), solitary fibrous tumor (2 cases), schwannoma (1 case), and diffuse large B cell lymphoma (1 case). Visual acuity and ocular motility were unchanged or improved in all cases. There were no visible scars or other complications related to this approach.

Conclusion

The upper fornix approach combined with a superior lateral cantholysis is a novel technique that provides safe and excellent exposure of the deep superonasal orbit. In addition, it avoids the unnecessary morbidity of upper lid splitting, medial rectus muscle detachment, or bone removal.

     

Twenty five-gauge endoscopic vitrectomy for proliferative vitreoretinopathy with severe corneal opacity

Purpose

To report 4 cases undergoing 25-gauge endoscopic vitrectomy for the treatment of proliferative vitreoretinopathy with severe corneal opacity in which a transpupillary view of the fundus was not possible.

Study Design

A retrospective interventional case series.

Methods

The main outcomes measured were postoperative anatomic status of the retina and subjective improvement of vision. Results: Postoperative reattachment of the retina and subjective improvement of vision were achieved in all 4 eyes.

Conclusion

Twenty five-gauge endoscopic vitrectomy provides a clear view making it possible conduct pars plana vitrectomy in order to reattach the retina in cases of proliferative vitreoretinopathy with severe corneal opacity.

     

Effect of Berberine on Barrier Function in a Human Retinal Pigment Epithelial Cell Line

Purpose

To examine the effects of berberine, an alkaloid isolated from some medicinal herbs, on the disruption of the barrier function in a human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1β (IL-1β).

Methods

ARPE-19 cells were cultured to confluence. Berberine and IL-1β were added to the medium. Barrier functions were evaluated by measuring transepithelial electrical resistance (TER) and the permeability to horseradish peroxidase (HRP) and sodium fluorescein (SF).

Results

Berberine dose-dependently inhibited decreased TER and increased the permeability to HRP and SF in the cells stimulated with IL-1β.

Conclusions

Berberine dose-dependently inhibited the disruption of the barrier function in the ARPE-19 cell line induced by IL-1β.?Jpn J Ophthalmol 2007;51:64–67 © Japanese Ophthalmological Society 2007

     

Near-infrared and short-wave autofluorescence in ocular specimens

Purpose

To determine histopathologic characteristics of near-infrared autofluorescence (NIR-AF) and short-wave autofluorescence (SW-AF) in ocular tissue.

Study design

Retrospective study.

Methods

Unstained specimens from four enucleated eyes with uveal melanoma were prepared for evaluation by fluorescence microscopy. The filter settings for SW-AF were 450-490 nm for excitation, 500-550 nm for emission and for NIR-AF 672.5-747.5 nm and 765-855 nm respectively.

Results

Hyper-SW-AF was detected in the cornea, crystalline lens, anterior border layer of the iris, basement membrane of the iris posterior epithelium, retinal pigment epithelium (RPE), Bruch’s membrane, and sclera. Hyper-NIR-AF was detected in pigmented tissues, i.e., iris anterior border layer, iris posterior epithelium, ciliary pigmented epithelium, RPE, pigmented cells in the choroid and pigmented cells in the melanoma tumoral masses. The iris anterior border layer had hyper-SW-AF and hyper-NIR-AF with low magnification. The cells on the iris surface were with hyper-SW-AF; under the iris surface cells with hyper-NIR-AF were detected with high magnification. Both hyper-SW-AF and hyper-NIR-AF were in RPE cells. Pigmented cells with hyper-NIR-AF in other uveal tissues did not have hyper-SW-AF. The pigmented cells in the melanoma tumoral masses had very weak NIR-AF.

Conclusions

NIR-AF was seen in the ocular pigmented tissues. The only pigmented tissue with both hyper-SW-AF and hyper-NIR-AF was RPE, the combination of which might help interpret the cellular components of fundus lesions.

     

Pseudotumor cerebri associated with Sjögren's syndrome

Purpose

To report a case of papilledema and pseudotumor cerebri developed in association with Sjögren's syndrome.

Methods

Case-report of a 38-year-old woman with history, imaging and histology confirming the diagnosis of both pseudotumor cerebri and Sjögren's syndrome who presented with bilateral decrease of vision.

Results

Papilledema associated with pseudotumor cerebri was observed in both eyes. The patient's visual acuity improved transiently with the administration of intravenous steroids and cyclophosphamide; subsequently she needed a ventriculoperitoneal shunt.

Conclusion

Sjögren's syndrome should be considered in the different etiologies of pseudotumor cerebri. The major improvement with corticosteroids and ventriculoperitoneal shunt makes prompt diagnosis essential.

     

Advanced glaucoma following Nd:YAG capsulotomy: a case report

Background

We present a case of aphakic pupil block caused by vitreous prolapse into the anterior chamber following Nd:YAG capsulotomy.

Case presentation

This resulted in advanced glaucoma in a young patient, which presented a significant clinical management challenge.

Conclusions

Ultimately, at the time of writing, her intraocular pressure and uveitis were well controlled, however the long-term outcome remains uncertain, given the uncompromising natural history of her complicated ocular condition.

     

Effect of laser peripheral iridotomy using argon and neodymium-YAG lasers on corneal endothelial cell density: 7-year longitudinal evaluation

Purpose

To evaluate the changes in corneal endothelial cell density (ECD) over a 7-year period after laser peripheral iridotomy (LPI) using argon and neodymium-doped yttrium aluminum garnet (Nd:YAG) lasers.

Study design

Retrospective case series.

Methods

Eyes that underwent prophylactic LPI using argon and Nd:YAG lasers were followed up for 7 years. Central corneal endothelial cells were observed by use of noncontact specular microscopy preoperatively and at 1 and 7 years postoperatively. Changes in ECD and the associations between preoperative ECD and the total energy of the Nd:YAG laser were evaluated.

Results

Fifty-one eyes of 51 patients were followed up for 7 years. The ECD significantly decreased after LPI (P < 0.049), and the reduction rate at 1 year after the surgery (1.69 ± 4.80%, 95% CI: 0.34%–3.04%) was significantly higher than the annual reduction rates after 1 year (0.17 ± 0.85%/y, P = 0.036, 95% CI: -0.07% to 0.41%). No association was found between the preoperative ECD and the total laser energy.

Conclusions

Long-term evaluation indicated that the reduction in ECD after argon-Nd:YAG laser LPI was present but small during the initial year and was negligible after 1 year.

     

Evaluating the growth of eyelid lesions: comparison of clinical evaluation and a software-based model

Purpose

To evaluate the accuracy of clinicians in evaluating the growth of eyelid lesions and to compare the measurements of experienced ophthalmologists to a novel computerized measurement method.

Design

Prospective, single center, observational study.

Methods

Six experienced ophthalmologists were asked to measure 3 simulated eyelid lesions using a slit lamp. These lesions were then enlarged, and the same examiners were asked to measure the enlarged lesions without prompting that the lesions had changed. Slit lamp photography of the original lesions and enlarged lesions were analyzed using freely available software from the National Institutes of Health. The results of clinician measurements were compared to the software-generated data.

Results

Clinician data regarding the growth of the lesions were as follows: ?40.9 to +76.8% for lesion 1, +29.3 to +134.4% for lesion 2, and +148.5 to +1169.2% for lesion 3. Software-based measurements were as follows: +53.6, +100.7, and +182.2% for lesions 1, 2, and 3, respectively.

Conclusions

Monitoring growth of eyelid lesions in clinical practice can be challenging. We propose that using computerized software to analyze surface area of concerning eyelid lesions may provide a significant advantage over current clinical practices.

     

Blood coagulation parameters after intravitreal injection of aflibercept in patients with neovascular age-related macular degeneration

Purpose

The aim of this study was to evaluate the effect of intravitreal injection of aflibercept (IVA) on blood coagulation tests in neovascular age-related macular degeneration (AMD) patients.

Methods

Thirty-four patients with neovascular AMD (study group) and 32 healthy individuals (control group) were enrolled. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were measured at different times in patients with neovascular AMD.

Results

The levels of PT and aPTT after IVA were decreased at 1 month after the first injection and 1 month after the second injection compared to the baseline measurement in the study group.

Conclusions

IVA may cause a decrease in the levels of PT and aPTT at 1 month after the first injection and 1 month after the second injection although these results are not statistically significant in our study.

     

Frequencies of human leukocyte antigen alleles and haplotypes among Japanese patients with age-related macular degeneration

Purpose

Stem cell therapy is a potential treatment for retinal disorders. We are currently exploring treating HLA matched patients of age-related macular degeneration (AMD) by using allogenic retinal pigment epithelium cells derived from induced pluripotent stem cells (iPS-RPE) from human leukocyte antigen (HLA) homozygote donors. The purpose of this study was to investigate the frequency of HLA class I and II alleles and haplotypes in Japanese patients with AMD.

Study design

Cross-sectional observation clinical study.

Methods

A total of 138 consecutive patients diagnosed with neovascular AMD (mean age, 76.0?±?7.8 years, 105 men) and 300 controls were included in the study. The frequencies of HLA-A, -B, -C, -DRB1, -DQB1, and -DPB1 alleles were determined using illumina MiSeq platform. Frequencies of HLA alleles at six loci in patients with AMD were compared with those of the controls.

Results

The alleles with the highest prevalence at each locus were A*24:02 (29.7%), B*52:01 (15.5%), C*12:02 (16.1%), DRB1*09:01 (19.1%), DQB1*06:01 (23.2%), and DPB1* 05:01 (40.5%). There were no significant associations between the HLA alleles and AMD. The most common haplotype was A*24:02-B*52:01-C*12:02-DRB1*15:02-DQB1*06:01-DPB1*09:01, with a 9.8% genetic frequency among all haplotypes, detected in 18.8% of the patients.

Conclusion

The genotype of HLA in patients with AMD was not different from that in the Japanese control population. Thus, therapy with iPS-RPEof the most frequent HLA haplotype could be a feasible alternative for AMD in a wider population.

     

Morphometric features on enhanced depth imaging optical coherence tomography scans in idiopathic posterior uveitis or panuveitis

Purpose

Enhanced depth imaging (EDI) optical coherence tomography (OCT) has emerged as a novel tool for qualitative and quantitative choroidal assessment in posterior uveitis. The objective of this study was to investigate the role of EDI-OCT to assess the choroidal and retinal changes in posterior uveitis.

Methods

In this retrospective study, EDI-OCT scans of patients with idiopathic posterior uveitis or panuveitis were reviewed. Morphological features from retina and choroid from the OCT scans were assessed and compared to the fellow normal eyes. Follow-up assessment was performed at 6-month follow-up.

Results

Nineteen patients with idiopathic posterior or panuveitis were included in the study. Choroidal examination using EDI-OCT scans showed areas of focal hypo-reflective and discrete hyper-reflective foci in one-third patients. Macular edema, disruption of the ellipsoid zone (generalized and discrete), outer retinal hyper-reflective foci, and intraretinal and subretinal fluid were observed.

Conclusions

The index study reports qualitative OCT-derived parameters as possible tools in monitoring disease progression in uveitis.

     

The influence of mitomycin C concentration on the outcome of trabeculectomy in uveitic glaucoma

Purpose

To evaluate the influence of mitomycin C (MMC) concentration on the outcome of trabeculectomy in uveitic glaucoma.

Methods

Retrospective comparative study included 50 patients who underwent MMC-enhanced trabeculectomy for uncontrolled uveitic glaucoma. Patients were divided into two groups based on the concentration of MMC used during trabeculectomy (high [0.04%] or low [0.02%]). The main outcome measures were: the intraocular pressure (IOP), the number of antiglaucoma medications, the need for further glaucoma surgeries, and trabeculectomy success.

Results

There were no differences in the IOP nor antiglaucoma medications. The success rates were 76.0 and 68.0%, while the failure rates were 24.0 and 32.0% in the 0.02 and 0.04% MMC groups, respectively.

Conclusion

Trabeculectomy augmented with low (0.02%) MMC concentration seems to have comparable outcomes in uveitic glaucoma compared with high (0.04%) concentration.