Ophthalmic Research (OPHTHAL RES )

Publisher: Blackwell Publishing


ëOphthalmic Researchí features original papers, reviews and short communications reporting basic and clinical experimental studies. Authors from throughout the world cover morphologic, physical, physiologic, pharmacological, biochemical and molecular biological aspects of ophthalmology and experimental eye research. Articles on methodological problems are included as well. The results of new experimental research are also interpreted in light of their importance to the clinical work of the eye specialist. This journal provides a record of international research for both researchers and clinicians in ophthalmology.

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Publications in this journal

  • [Show abstract] [Hide abstract]
    ABSTRACT: Objective: To report the clinical outcome of autologous cultured limbal epithelial cell transplantation (CLECT) followed by deep anterior lamellar keratoplasty (DALK) in paediatric eyes and to correlate the clinical outcome with the phenotype of rejuvenated corneal epithelium. Methods: Four patients with total limbal stem cell deficiency (LSCD) underwent autologous CLECT. Cultivated cell sheets were transplanted onto the damaged ocular surface followed by DALK surgery. Excised corneal buttons were subjected to histopathological analysis. Data recorded included age, sex, laterality, nature of injury, follow-up period, severity of stem cell deficiency, visual acuity, Schirmer's test and impression cytology. Results: At a mean follow-up period of 19.5 ± 7.4 (range 9-26) months after CLECT, all 4 eyes showed epithelialized and clinically stabilized ocular surface. Manual DALK was performed in all 4 eyes, with a mean follow-up of 9.75 ± 4.5 (range 5-15) months. All eyes exhibited smooth and clear corneal epithelium with improved visual acuity. Excised corneal buttons demonstrated organized corneal epithelial morphology and showed expression of cornea-specific CK3/12 marker. Conclusion: Restoration of severely damaged ocular surface following chemical injury by using 2-stage meticulous approaches offers a new modality for the treatment of severe LSCD. Transplantation of cultivated autologous limbal epithelial cell sheet followed by DALK surgery can efficiently restore the corneal phenotype with improved vision.
    Ophthalmic Research 06/2013; 50(1):59-64.
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    ABSTRACT: Background: Epithelial basement membrane dystrophy (EBMD) is by far the most common corneal dystrophy. In this study, we used a newly developed method of immunofluorescence staining and imaging to study the entire corneal nerve architecture of a donor with unilateral EBMD. Method: Two fresh eyes from a 56-year-old male donor were obtained; the right eye of the donor was diagnosed with EBMD and the left was normal. After slit lamp examination, the corneas were immunostained with anti-β-tubulin III antibody. Images were recorded by a fluorescent microscope equipped with a Photometrics digital camera using MetaVue imaging software. Results: The left cornea appeared normal as observed by slit lamp and stereomicroscope, but the right eye had numerous irregular geographic patches in the basement membrane. Immunofluorescence showed no difference in the stromal nerve distribution between the 2 eyes, but there were areas without innervations in the EBMD cornea. Subbasal nerve fibers also showed tortuous courses and fewer divisions. There was a significant decrease in the density of subbasal nerve fibers and the number of terminals in the right eye. Conclusion: We show for the first time detailed nerve architecture in an EBMD cornea. Our results suggest that EBMD-induced abnormalities of basement membrane altered epithelial nerve architecture and decreased nerve density, contributing to the pathology of the disease.
    Ophthalmic Research 01/2013; 49(4):185-191.
  • Ophthalmic Research 01/2013; 50(2):99-107.
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    ABSTRACT: RhoA is a small guanosine triphosphatase which participates in signaling pathways of axonal repellents or inhibitors. However, the distribution and expression of RhoA in the rat retina after optic nerve injury has not been elucidated yet. To study the distribution and expression of RhoA in the rat retina after optic nerve injury. Immunohistochemistry was used to determine the distribution of RhoA in rat retina after optic nerve injury. The expression of RhoA was analyzed by Western blot. In normal retina and the retina 1 day after optic nerve injury, RhoA was distributed in the retinal ganglion cell (RGC) layer. Three days after optic nerve injury, it existed in RGCs and the inner plexiform layer. However, 7 days after surgery its immunoreactivity was abundant not only in the RGC and inner plexiform layers but also in the inner nuclear and outer plexiform layers. Western blot analysis showed that the expression of RhoA increased significantly in the retina after optic nerve injury in comparison with normal retina. These results indicate that the distribution and expression of RhoA were extended and enhanced after optic nerve injury, and that RhoA plays an important role in optic nerve regeneration.
    Ophthalmic Research 02/2007; 39(3):174-8.
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    ABSTRACT: To report detection of leprosy in ocular tissue by histopathology and its confirmation by genetic analysis. Excised tissue from a clinically-suspected ocular leprosy patient was processed and analyzed histopathologically. The DNA from the paraffin-embedded tissue was extracted, an 85 A-C intergenic region of Mycobacterium leprae was amplified using specific primers and analyzed by conventional as well as real-time polymerase chain reaction (RT-PCR). With periodic acid-Schiff-hematoxylin (PAS-H) staining the specimen showed presence of a thin fibrinous layer of inflammatory cells. The majority of the tissue was fibrovascular with extensive infiltration by histiocytes having reticulated cytoplasm. Modified PAS-H and acid-fast staining (AFS) showed the presence of several acid-fast organisms within the cytoplasm of histiocytes and mast cells. Conventional PCR showed a 250-bp DNA from excised conjunctival tissue, which was in agreement with the positive controls for M. leprae. Through RT-PCR, it was calculated that the suspected tissue had 44.68 pg of M. leprae DNA, which is 8937.06 genome copies of M. leprae. Presence of inflammatory cells and AFS bacilli in tissue presented a typical picture of leprosy. M. leprae DNA can be detected using RT-PCR in ocular tissues when acid-fast bacteria are seen in histopathological sections. And when the diagnosis of leprosy is inconclusive and acid-fast bacteria are seen, RT-PCR for M. leprae DNA could be used as a rapid confirmatory test to identify the presence of M. leprae and, therefore, the diagnosis of leprosy.
    Ophthalmic Research 02/2007; 39(2):63-8.
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    ABSTRACT: Retinal pigment epithelial cells (RPE cells) are key players in the first-line defense against invading organisms such as viruses and bacteria. The interaction between RPE cells and viral or bacterial components is very important for clearance of these organisms. Toll-like receptors are a family of recognition receptors involved in innate immunity. Each TLR acts as a primary sensor of conserved microbial components and drives the induction of specific biological responses. TLR 3 is involved in the recognition of viral components, such as double-stranded RNA (dsRNA) and poly(I:C), while TLR 9 recognizes viral or bacterial DNA without methylation at CpG motifs. In the present study, we investigated the expression and function of TLR 3 and 9 in RPE cells. PCR analysis revealed expression of genes for TLR 3 and 9 in RPE cells. Expression of TLR 3 and 9 protein was detected in RPE cells by flow cytometry. TLR 3 and 9 showed strong intracellular expression. To detect angiogenetic factors produced by RPE cells, culture supernatant was examined with the Human Angiogenesis Antibody Array, which can simultaneously detect 20 different angiogenetic factors including cytokines, chemokines, soluble cytokine receptors, and growth factors. RPE cells showed high production of interleukin-8 (IL-8) and monocyte chemotactic protein-I (MCP-I). Furthermore, stimulation of RPE cells with the dsRNA analogue poly(I:C) enhanced the secretion of IL-8 and MCP-I, as well as enhancing the expression of junctional adhesion molecule-I (Jam-I) and intracellular adhesion molecule-I (ICAM-I), and promoted the adhesion of monocyte to these cells. In contrast, stimulation with the CpG-DNA motif only enhanced the secretion of IL-8. However, CpG-DNA motif enhanced phagocytosis in RPE cells. These results may indicate that TLR 3 and 9 play a distinct role in the inflammatory response that clears viruses from the retina.
    Ophthalmic Research 02/2007; 39(3):155-63.
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    ABSTRACT: To evaluate the histological subtypes of basal cell carcinoma (BCC) of the eyelid and to determine their effect on the size, depth of invasion and need of retreatment of a nonselected patient material seen in south-western Finland. We studied the case records and the histological characteristics of BCC of the eyelid treated at the Turku University Eye Clinic during the years 1988 through 1997. The material consisted 103 patients (103 BCC tumors of the eyelid). All tumors were surgically excised. Histological slides were reviewed by a pathologist and the material was divided into histopathological subtypes. In 78.3% of the cases, the diameter of the lesion was smaller than 10 mm. The most frequent histological subtype was nodular (84.5%) followed by sclerosing (5.8%), micronodular (4.9%), keratotic (2.9%) and superficial (1.9%) types of BCC of the eyelid. Only patients of the nodular subtype showed recurrences (11 cases). The size of the tumor and the depth of invasion correlated directly with each other. However, some nodular types of BCC tumors smaller than 10 mm in diameter extended to a depth of more than 4.0 mm. The nodular subtype of BCC should be regarded as a potentially invasive and recurrent tumor. Histopathological examination and subtyping of all BCC tumors is recommended.
    Ophthalmic Research 02/2007; 39(1):45-8.
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    ABSTRACT: It was the aim of this study to report on the intravitreal use of bevacizumab as antiproliferative agent in combination with filtering surgery. The clinical interventional case series study included 2 patients (2 eyes) who underwent standard antiglaucomatous penetrating filtering surgery combined with an intravitreal application of 1.5 mg bevacizumab. The intraocular pressure was elevated due to an intravitreal triamcinolone injection as treatment of exudative age-related macular degeneration (patient No. 1) or due to neovascular glaucoma (patient No. 2) after an ischemic retinal branch vein occlusion. At 4 and 12 weeks after surgery, intraocular pressure was reduced in both patients to 10 and 14 mm Hg with functioning filtering blebs. Intravitreal bevacizumab may potentially be helpful as addition to antiglaucomatous filtering surgery, particularly in neovascular glaucoma.
    Ophthalmic Research 02/2007; 39(2):121-2.
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    ABSTRACT: To evaluate the concentrations of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in neovascular or edematous retinal diseases. In the clinical comparative interventional study, VEGF and bFGF concentrations in aqueous humor samples of 35 patients with exudative age-related macular degeneration (AMD), 21 patients with diabetic macular edema and 24 patients of a control group were measured using a solid-phase chemiluminescence immunoassay. Concentrations of VEGF and bFGF, respectively, were significantly higher in the diabetic group (184.7 +/- 107.0 and 5.0 +/- 10.2 pg/l) than in the AMD group (107.7 +/- 73.0 pg/l, p = 0.002; 2.2 +/- 7.4 pg/l, p = 0.002) and the control group (71.5 +/- 94.7 pg/l, p = 0.001; 0.00 pg/l, p = 0.001). The two latter groups did not vary significantly (p = 0.10). VEGF and bFGF are present in considerably higher concentrations in eyes with diabetic macular edema than in eyes with exudative AMD or normal eyes. The differences were more marked for VEGF than for bFGF.
    Ophthalmic Research 02/2007; 39(3):139-42.
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    ABSTRACT: To perform lamellar keratolimbal allograft transplantation in a one-step procedure with a single graft, we investigated the feasibility of harvesting eccentric lamellar keratolimbal grafts from conventionally processed corneoscleral buttons using a manually guided microkeratome in conjunction with an artificial anterior chamber system. We used the Moria LSK-One microkeratome and the automated lamellar therapeutic keratoplasty (ALTK) system (Antony, France). Ten human donor eyes were used to obtain single-piece lamellar keratolimbal grafts. Specimens were processed for light and electron microscopy. Eccentric keratolimbal grafts could be obtained from all human donor buttons. Grafts include a crescent-shaped limbal and a large corneal portion. No visible damage to the limbal region was discernible. Our data show that the LSK-One microkeratome in conjunction with the ALTK system allows harvesting eccentric keratolimbal grafts from donor corneoscleral buttons.
    Ophthalmic Research 02/2007; 39(3):179-83.
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    ABSTRACT: To study the role of CD4+ natural killer T (NKT) cells in staphylococcal-enterotoxin-B (SEB)-treated rats after high-risk corneal transplantation. Fisher 344 donor corneas were transplanted into Lewis recipients. Corneal neovascularization was induced by sutures. All the recipients were randomly divided into 3 groups. The SEB group was intraperitoneally injected with SEB at a concentration of 75 microg/kg. The drug combination group received SEB and dexamethasone at a concentration of 5 mg/ml. The control group received saline buffer. All transplants were evaluated for 30 days. Ten days after transplantation, 3 recipients in each group were sacrificed for immunological study. The survival time of the allografts in the SEB group was 12.50 +/- 1.41 days, much longer than in the control group (7.30 +/- 0.67 days) and the drug combination group (10.38 +/- 3.07 days). The lymphocyte proliferation ability was the weakest and the percentage of CD4+ NKT cells in both the spleen and the mandibular lymph nodes was the highest in the SEB group, while the percentage of CD4+ and CD8+ cells was the lowest in the drug combination group. IL-2 in the aqueous humor and the serum was lower while IL-10 was higher in the SEB group than in the other 2 groups. SEB prolongs allograft survival in rat high-risk corneal transplantation. This effect seems to be mediated by the upregulation of CD4+ NKT cells.
    Ophthalmic Research 02/2007; 39(3):130-8.
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    ABSTRACT: In glaucoma, retinal ganglion cell (RGC) death is induced by many risk factors, including ocular hypertension. It has been proposed that glutamate-mediated oxidative stress may also contribute to this RGC death. Cannabinoids are known to possess therapeutic properties including ocular hypotension and antioxidation. In this study, we test the hypothesis that (-)Delta(9)-tetrahydrocannabinol (THC) lowers intraocular pressure (IOP) and prevents RGC death in a rat model of glaucoma. Arat model of experimental glaucoma with chronic, moderately elevated IOP was produced unilaterally by cauterization of episcleral vessels. Rats received weekly injections of THC at a level of 5 mg/kg or vehicle for 20 weeks. IOP of both eyes was measured weekly on anesthetized animals immediately before THC treatment. RGCs were labeled in a retrograde fashion and counted in whole-mounted retinas. IOP was elevated in all operated eyes 1 day after the operation and remained elevated in the vehicle-treated rats throughout 20 weeks. In THC-treated rats, IOP elevation in operated eyes was diminished 2 weeks after operation and remained reduced. IOP in the contralateral control eyes was not affected by THC. In the operated eyes of vehicle-treated animals, there was a loss of approximately 50 and 40% of the RGCs in the peripheral and central retina, respectively. The RGC loss in the operated eyes of the THC-treated animals was reduced to 10-20%. These results demonstrate that THC is a neuroprotectant that preserves RGCs in an experimental model of glaucoma, possibly through a reduction in IOP.
    Ophthalmic Research 02/2007; 39(2):69-75.
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    ABSTRACT: To compare the efficacy ofa combination therapy of triamcinolone acetonide (TA) administration with and without vitrectomy in eyes with macular edema associated with branch retinal vein occlusion over a 1-year period. A retrospective, case-control study was conducted in 15 eyes of 15 patients with macular edema associated with branch retinal vein occlusion. Eight eyes underwent simultaneous intravitreal and posterior sub-Tenon capsule injections of TA (TA-injected group). Seven eyes underwent vitrectomy with intravitreal or simultaneous posterior sub-Tenon capsule injection of TA (vitrectomy with TA group). Macular thickness and visual acuity were measured before and at 1, 3, 6 and 12 months after the therapy. Twelve months after the therapy, mean visual acuity improved significantly from baseline in both the TA-injected (p = 0.0069) and vitrectomy with TA groups (p = 0.0145). Macular thickness also improved significantly in both the TA-injected (p = 0.0065) and vitrectomy with TA groups (p = 0.0058). At 12 months after the therapy, there was no significant difference in visual acuity and macular thickness between the two groups (p = 0.3308 and 0.3711, respectively). At the early postoperative stage (1 and 3 months after the therapy), the central macular thickness in the TA-injected group was significantly less than that in the vitrectomy with TA group (p = 0.0140 and 0.0275, respectively); there was no significant difference in visual acuity between the two groups (p = 0.0796 and 0.3753, respectively). TA injection without vitrectomy was as effective as a combination therapy of TA injection with vitrectomy.
    Ophthalmic Research 02/2007; 39(4):207-12.
  • Ophthalmic Research 02/2007; 39(2):62.
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    ABSTRACT: Mutations and polymorphisms have been identified in the CYP1B1 gene; while mutations that affect the conserved core structures of cytochrome P4501B1 result in primary congenital glaucoma (PCG), mutations in other regions hold the potential to define differences in estrogen metabolism. In the present study, we analyzed the CYP1B1 gene in Mexican patients with PCG and described four novel mutations. The sample included 12 nonrelated cases with PCG. Analysis of coding regions of the CYP1B1 gene was performed through PCR and DNA sequencing analysis from genomic DNA. Molecular analysis of the CYP1B1 gene showed the following molecular defects: (1) a novel single-base pair deletion within codon 370 (1454delC) that produces a substitution of leucine instead of proline and a premature stop codon 57 amino acids after the last original amino acid; this family also harbored a novel polymorphic variant of the cytochrome P4501B1 with six single-nucleotide polymorphisms (142C-->G; 355G-->T; 729G-->C; 4326C-->G; 4360C-->G and 4379C-->T); (2) a novel single-base pair deletion within codon 277 (1176delT) that results in a premature stop codon; (3) a novel single-base pair deletion within codon 179 (880delG) that produces a substitution of arginine instead of alanine and a premature stop codon 17 amino acids downstream from the last original amino acid, and (4) a duplication (or insertion) of ten base pairs within codon 404 (1556dupATGCCACCAC) that results in a premature stop codon 26 amino acids after the last original amino acid. We also observed in 2 nonrelated patients a deletion of 13 bp (1410_1422delGAGTGCAGGCAGA) previously reported for other populations. We reported four novel mutations and a novel polymorphic variant in the CYP1B1 gene in PCG in the Mexican population; it has important implications in diagnosis and genetic counseling.
    Ophthalmic Research 02/2007; 39(1):17-23.
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    ABSTRACT: The immunohistochemical expressions of two multidrug resistance proteins, P-glycoprotein (P-gp) and multidrug resistance-related protein-1 (MRP-1), were studied in retinoblastoma and the correlations with the clinicopathological parameters were assessed. Sixty-five enucleated eyes containing retinoblastoma were included in the study. Following hematoxylin-eosin staining, tumor differentiation, presence of choroidal invasion, optic nerve invasion, retinal invasion, necrosis and presence of calcification were evaluated with the light microscope. P-gp and MRP-1 expressions were evaluated immunohistochemically. Fifty-three eyes were enucleated primarily and 12 eyes were operated after failure of chemotherapy. P-gp and MRP-1 expressions were positive in 69.3 and 73.4% of specimens, respectively. There was no statistically significant relationship between the expressions of P-gp and MRP-1, and tumor differentiation, presence of tumor invasion or treatment with chemotherapy. Retinoblastoma intrinsically expresses both P-gp and MRP-1 and their expressions are not related to tumor differentiation. The expressions of P-gp and MRP-1 do not seem to be induced by chemotherapy and are not related to the degree of tumor invasion.
    Ophthalmic Research 02/2007; 39(4):191-7.
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    ABSTRACT: To study the interaction between latanoprost and pilocarpine on cultured rabbit ciliary muscle (RCM) cells, and investigate the time courses of the two drugs, when given alone or in combination. Cultured RCM cells were treated for 24 h with different concentrations of latanoprost acid, pilocarpine and mixtures of latanoprost acid and pilocarpine. RNA was extracted, expressions of matrix metalloproteinase 1 (MMP-1), tissue inhibitor of metalloproteinase 1 (TIMP-1) and TIMP-2 were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the optimum concentrations of those drugs were found. Then the cells were treated with the optimum concentrations of those drugs for various periods. RNA was extracted after the treatment and expressions of MMP-1, TIMP-1 and TIMP-2 were detected by RT-PCR again. Changes in [Ca(2+)](i) were estimated by fluorescence measurement using the Ca(2+) indicator Fluo-3 AM with a laser scanning confocal microscope. [Ca(2+)](i) of each cell was monitored continually after administration of the drugs. Gray values at 5 s and 2, 4, 6, 8 and 10 min were chosen for statistical analysis, and the influence and time-effect relationship of those drugs on [Ca(2+)](i) of the cultured cells were evaluated. Exposure of the cells to increasing concentrations of latanoprost acid induced increased MMP-1 mRNA and decreased TIMP-1 and TIMP-2 mRNA in a dose-dependent manner. After 24 h of treatment, the optimum concentration of latanoprost acid for maximal changes in MMP-1 and TIMP-2 expression was 2 x 10(-7)M, and for maximal changes in TIMP-1 expression, the optimum concentration was 5 x 10(-7)M. When the optimum concentrations of latanoprost acid were chosen to treat the cells for various periods, the optimum time of the peak MMP-1 expression and trough TIMP-1 expression was 24 h, and of the trough TIMP-2 expression, it was 36 h after initiation of treatment. No significant expression changes of MMP-1, TIMP-1 and TIMP-2 mRNA were found when the cells were treated with pilocarpine at any concentration or at any time. Exposure of the cells to the mixtures of latanoprost acid and pilocarpine induced the same changes and time course of MMP-1, TIMP-1, and TIMP-2 mRNA expression as exposure of the cells to latanoprost acid alone. Exposure of ciliary muscle cells to pilocarpine induced an increase in [Ca(2+)](i), with the peak of increase observed at 5 s after initiation of treatment; then [Ca(2+)](i) gradually decreased near to baseline level within 10 min. Exposure of the cells to latanoprost acid did not significantly change [Ca(2+)](i). Exposure of the cells to the mixtures of latanoprost acid and pilocarpine induced the same [Ca(2+)](i) change as exposure to pilocarpine alone. Latanoprost and pilocarpine have no interaction in their various effects on the cultured RCM cells.
    Ophthalmic Research 02/2007; 39(4):232-40.
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    ABSTRACT: In order to characterize the biological effects and molecular mechanism underlying indocyanine-green (ICG)-mediated photo-oxidative cell damage, human cultured retinal pigmented epithelium (RPE) cells preloaded with ICG were exposed to 810-nm laser irradiation. Cell viability and death induction were examined, as well as the modulation of proteins involved in cell death and DNA repair. ARPE-19 cells preloaded with 100 microM ICG were irradiated using continuous and micropulsed 810-nm laser for the dye photoactivation, and cell viability and apoptosis were evaluated. The expression and subcellular localization of Bax, Ku70, Ku80 and clusterin/ApoJ were analyzed by immunocytochemistry and Western blot. ICG photoactivation induced apoptosis in RPE cells. The micropulsed laser irradiation induced a higher percentage of cell killing as compared to continuous wave. Cell killing was inhibited by sodium azide, suggesting the involvement of reactive oxygen species in the laser-induced cell damage. Bax was strongly induced after 4 and up to 24 h of treatment. The nuclear proapoptotic isoform of clusterin/ApoJ was selectively upregulated after 24 h of treatment. The DNA repair machinery was upregulated after 4 and up to 24 h. These data elucidate some molecular mechanisms involved in cell death induced by ICG photosensitization. The increase and relocalization of Bax into the mitochondria and the upregulation and translocation of the proapoptotic isoform of clusterin/ApoJ in the nucleus demonstrated the involvement of these proteins in the photo-oxidative cell death pathway. These data point out new molecular targets and suggest potential applications in the therapy of the retinal diseases that could benefit by selective RPE treatment.
    Ophthalmic Research 02/2007; 39(3):164-73.
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    ABSTRACT: Cyclic 3',5'-guanosine monophosphate (cGMP), a central molecule in the phototransduction cascade, is also involved in a number of other physiological processes in the retina, like stimulating the absorption of subretinal fluid by activating the retinal pigment epithelium (RPE) cell pump. The aim of this study was to quantify cGMP synthesis by RPE cells and to investigate the role of two separate enzymatic pathways (soluble versus particulate guanylyl cyclase) in its production. cGMP expression was evaluated by immunochemistry and radioimmunoassay following culture of the D407 RPE cell line in the presence of a nonselective phosphodiesterase inhibitor (IBMX), in combination with the particulate guanylyl cyclase stimulator atrial natriuretic peptide (ANP) or the soluble guanylyl cyclase stimulator sodium nitroprusside (SNP). Stimulation of the particulate guanylyl cyclase in RPE cells with ANP resulted in high intra- and extracellular cGMP levels. Stimulation of the soluble guanylyl cyclase by SNP resulted in a slight elevation of cGMP levels compared to controls. These results show that cultured human RPE cells are capable of producing cGMP and that most cGMP is generated following stimulation of the particulate guanylyl cyclase pathway.
    Ophthalmic Research 02/2007; 39(1):55-9.