Blunt needle revision with viscoelastic materials via the anterior chamber for early failed filtering blebs after trabeculectomy.

PURPOSE: To report a new technique of blunt needle revision with viscoelastic materials via the anterior chamber for the treatment of early failed filtering blebs and elevated intraocular pressure after trabeculectomy, in which digital ocular massage and laser suture lysis have been ineffective. METHODS: A 27-gauge blunt needle attached to a syringe containing viscoelastic material was inserted into the anterior chamber from the inferior paracentesis. The needle tip was inserted into the subscleral flap space from the filtering fistula at the anterior chamber side, and the scleral flap was lifted bluntly. The needle tip was then inserted into the subconjunctival space where the viscoelastic agent was injected and the adhesion between the sclera and conjunctiva was separated bluntly. Blunt needle revision via the anterior chamber was performed 14 times in six eyes of six patients at Saitama Medical Center, Jichi Medical University from January 2007 to May 2009. All procedures were performed within 1 month after trabeculectomy. RESULTS: The intraocular pressure remained 21 mmHg or lower for more than 6 months in three of six eyes. Slight bleeding from the iris occurred in one of the 14 procedures, and hypotony (intraocular pressure below 5 mmHg) occurred in one of the 14 procedures. No serious complications developed. CONCLUSION: Blunt needle revision via the anterior chamber for early failed filtering blebs is a new, simple, and safe procedure.

Clarifying the mechanism of idiopathic macular hole development in fellow eyes using spectral-domain optical coherence tomography.

BACKGROUND: To clarify the mechanism of idiopathic macular hole development, we evaluated the vitreoretinal relationship in fellow eyes of those with a macular hole and normal eyes using spectral-domain optical coherence tomography. Thirty-one fellow eyes and 34 normal volunteer eyes without a posterior vitreous detachment (PVD) were included. RESULTS: WE CLASSIFIED SIX VITREOMACULAR RELATIONSHIPS: type 1, no PVD, five fellow eyes (16.1%) and nine control eyes (26.5%); type 2, shallow PVD with perifoveal vitreous attachment, seven fellow eyes (22.6%) and 19 control eyes (55.9%); type 3, shallow PVD with pinpoint foveal vitreous traction, seven fellow eyes (22.6%) and no control eyes (0%), type 4a; shallow PVD with a round defect in the posterior vitreous cortex over the perifoveal area with vitreous attachment to the perifoveal area, two fellow eyes (6.5%) and one control eye (2.9%); type 4b, shallow PVD with a round defect in the posterior vitreous cortex over the perifoveal area without vitreous attachment to the perifoveal area, no fellow eyes (0%) and one control eye (2.9%); type 5a, shallow PVD with no pseudo-operculum, no fellow eyes (0%) and four control eyes (11.8%); type 5b, shallow PVD with a pseudo-operculum, four fellow eyes (12.9%) and no control eyes (0%); and type 6, biomicroscopically relevant PVD, six fellow eyes (19.4%). CONCLUSION: Types 3 and 5b developed only in fellow eyes. Type 2 developed most often in normal eyes and seemed to cause less foveal stress. Type 3 may show the basic pathogenesis of macular holes. Progression of type 5b after type 3 induces abortion of developing macular holes.

Surgical results of pars plana vitrectomy combined with pars plana lensectomy with anterior capsule preservation, endophotocoagulation, and silicon oil tamponade for neovascular glaucoma.

PURPOSE: To report on pars plana vitrectomy (PPV) combined with pars plana lensectomy (PPL) with a preserved anterior capsule, panretinal endophotocoagulation (EPC) throughout the pars plana, and silicon oil (SO) tamponade (PPV + PPL + EPC + SO tamponade) for neovascular glaucoma (NVG). METHODS: Thirteen eyes with NVG were treated. Ten eyes also underwent SO removal and intraocular lens (IOL) implantation (SO removal + IOL). Intraocular pressure (IOP), number of medications, and visual acuity were evaluated at the first visit, immediately before and 3 months after the procedure, 3 months after SO removal + IOL, and 1 year after the procedure. RESULTS: At the first visit, immediately before and 3 months after the procedure, 3 months after SO removal + IOL, and 1 year after the procedure, the IOPs were 29 ± 19, 23 ± 12, 13 ± 5, 17 ± 10, and 17 ± 6 mmHg; numbers of medications, 0.7 ± 1.4, 2.1 ± 2.0, 0.6 ± 0.7, 1.2 ± 1.2, and 1.6 ± 1.6; and best-corrected visual acuities converted to logarithm of the minimum angle of resolution (BCVA logMAR), 0.96 ± 0.96, 1.27 ± 0.80, 1.67 ± 0.91, 1.37 ± 0.89, and 1.90 ± 1.44, respectively. No severe hypotony or phthisis bulbi developed within 1 year after the procedure. The success rates (IOP ≤ 21 mmHg and sustained light perception) were 92.3% after 3 months and 69.2% after 1 year. CONCLUSION: PPV + PPL + EPC + SO tamponade might have prevented acute increases of vascular endothelial growth factor and inflammatory cytokine production postoperatively and resulted in good vision in patients with NVG.

Successful treatment of young-onset adult T cell leukemia/lymphoma and preceding chronic refractory eczema and corneal injury by allogeneic hematopoietic stem cell transplantation.

Only some carriers of human T cell lymphotropic virus type I (HTLV-1) develop adult T cell leukemia/lymphoma (ATLL) after a long latency period, and an association has been reported between chronic refractory eczema, known as infective dermatitis, and young-onset ATLL. A 25-year-old female developed ATLL and underwent allogeneic hematopoietic stem cell transplantation (HSCT) in non-remission. She had chronic refractory eczema and corneal injury at the onset of ATLL. Remission of ATLL was achieved, and the HTLV-1 proviral load decreased after HSCT. In addition, her pre-existing eczema and corneal injuries almost disappeared. More than a year has passed since the transplantation was performed, and she has had no recurrence of either ATLL or lesions in the skin and eye. Her clinical course suggests a possible association between skin and eye lesions and HTLV-1 infection. Changes in the immunological condition after HSCT might play a key role. Special attention is needed when HTLV-1 carriers develop eye or skin lesions.

Characteristics of diabetic retinopathy in SDT rats.

BACKGROUND: We previously reported a new diabetic strain of the Sprague-Dawley rat, named the Spontaneously Diabetic Torii (SDT) rat. The purpose of the present study was to report the histologic and ultrastructural characteristics of diabetic retinopathy (DR) in a new animal model, the SDT rat. METHODS: Fifty-three eyes of 43 SDT rats of various ages (35-82 weeks) were examined, of which 33 underwent histopathologic examination, 15 eyes fluorescein-dextran microscopy, and five eyes the trypsin digestion method. RESULTS: Of the 33 eyes examined histopathologically, DR was identified in 20 eyes (61%). Large retinal folds mimicking diabetic tractional retinal detachment were observed in 20 eyes (61%). Retinal hemorrhages were seen in four eyes (12%). A neovascular fibrous membrane around the iris developed in five eyes (15%), of which two eyes had a massive anterior chamber hemorrhage. Of the 15 eyes examined by fluorescein-dextran microscopy, an area of nonperfusion and/or extensive hyperfluorescence was observed in 12 eyes (80%). Of the five eyes examined using the trypsin digestion method, acellular capillaries and pericyte loss were observed in four eyes (80%). Of the 53 eyes, the previously mentioned retinal changes of DR were observed in 36 eyes (68%). The rats with DR (49-82 weeks; mean, 60 weeks) were older than the rats without DR (35-55 weeks; mean, 40 weeks) (p < 0.001). CONCLUSION: Large retinal folds mimicking tractional retinal detachment with extensive leakage of fluorescein around the optic disk was the most prominent finding of DR in SDT rats.

Sex-related effect on gene expression in the mouse meibomian gland.

PURPOSE: Sex-related differences have been identified in the anatomy and physiology of the meibomian gland. We hypothesize that these differences are due, at least in part, to variations in gene expression. This study's objective was to determine whether sex-related differences do exist in meibomian gland gene expression. We also sought to elucidate whether such differences, if any, might be (a) analogous to those known to occur in the lacrimal gland and (b) due to the effect of sex steroids. METHODS: Meibomian glands were obtained from young adult male and female BALB/c mice (n=7 to 15 mice per sex per experiment), pooled according to sex and processed for the isolation of RNA. Samples were evaluated for differentially expressed mRNAs by using CodeLink Bioarrays and GEM 1 and 2 gene chips. Bioarray data were analyzed with GeneSifter. Net software and also compared with microarray data in GEO and GeneSifter databases. RESULTS: Our results demonstrate that sex has a significant influence on the expression of 164 genes in the mouse meibomian gland. These genes are involved in a broad spectrum of biological processes, molecular functions, and cellular components, including such activities as metabolism, catalysis, cell growth and maintenance, membrane architecture, nucleic acid binding, transcription, and signal transduction. In addition, the nature of the sex-related variations in meibomian gland gene expression is quite different from those in the lacrimal gland and appear to be mediated in part by the action of androgens, but not estrogens or progestins. CONCLUSIONS: These findings support our hypothesis that sex-related differences exist in gene expression of the meibomian gland.

Influence of sex on gene expression in the mouse lacrimal gland.

Significant, sex-associated differences exist in the physiology and pathophysiology of the lacrimal gland. We hypothesize that many of these differences are due to fundamental variations in gene expression. The purpose of this study was to determine the extent to which sex-related differences in gene expression are present in the lacrimal gland. Lacrimal glands were obtained from adult male and female BALB/c mice (n=5-10mice/sex/experiment), pooled according to sex and processed for the isolation of RNA. Samples were analyzed for differentially expressed mRNAs by using Atlas Mouse cDNA Expression Arrays, cDNA amplification techniques, GEM 1 and 2 gene chips, CodeLink bioarrays and quantitative real-time PCR (qPCR) procedures. Quantitative evaluation of Atlas Array gene expression was performed with an image analysis system developed in our laboratory, whereas gene chip data were analyzed with Rosetta Resolver and GeneSifter.Net software. Statistical significance was determined by using Student's t-test. Our results with CodeLink bioarrays show that sex has a significant influence on the expression of over 490 genes in the mouse lacrimal gland. These genes are involved in a wide range of biological processes, molecular functions and cellular components, including such activities as development, growth, transcription, metabolism, signal transduction, transport, receptor activity and protein and nucleic acid binding. The expression of selected genes was confirmed by the use of GEM gene chips and qPCR. Our findings also demonstrate that certain methodological approaches are less useful in attempting to assess the magnitude of sex-associated differences in the lacrimal gland. These results support our hypothesis that sex-related differences in gene expression play a role in the sexual dimorphism of the lacrimal gland.

Distinct populations of dendritic cells in the normal human donor corneal epithelium.

PURPOSE: To characterize dendritic cells (DC) in normal human corneal epithelium. METHODS: Normal human donor corneal epithelium was examined by fluorescence microscopy with single and double staining for multiple markers. Morphologic studies were also performed by confocal microscopy. HLA-DRa, CD1c, and CD16 mRNA expression in the corneal epithelium was examined by RT-PCR. CD45+ cells were separated from the corneal epithelium with magnetic beads and then were stimulated with TNF-alpha and lipopolysaccharide in vitro. RESULTS: CD45+ cells were mainly located in the basal-cell layer of the corneal epithelium and partly in the wing/surface layers. CD45-positive cell numbers were significantly higher in the peripheral cornea (3-6 mm from the center) than in the central cornea (0-3 mm from the center). All these cells expressed HLA-DR and CD11c but not CD3, CD11b, CD14, CD19, CD56, or CD66, suggesting that these were bone marrow-derived myeloid DC. Some DR+CD11c+ DCs from the periphery expressed CD1c and CD16. HLA-DRa, CD1c, and CD16 mRNAs were detected in normal corneal epithelium. These CD11c+ DCs did not express CD123, CD1a, DC marker (CMRF56), CD40, CD80, or CD86. When CD45+ cells were isolated from the corneal epithelium by magnetic cell sorting, CD40 and CD86 expression were detected after in vitro stimulation with TNF-alpha and lipopolysaccharide. CONCLUSIONS: These findings demonstrate that normal human corneal epithelium contains at least three DC phenotypes, with HLA-DR+ myeloid lineage CD11c+CD16- DCs as the main population plus a small number of CD11c+CD16+ DCs and CD11c+CD1c+ DCs. These cells can be discriminated from bone marrow-derived cells in the human corneal stroma.

Androgen regulation of gene expression in the mouse lacrimal gland.

The objective of this study was to determine the nature and extent of androgen influence on gene expression in the lacrimal gland. Lacrimal glands were obtained from orchiectomized mice that had been treated with testosterone or vehicle for 2 weeks, as well as from testicular feminized mice and their Tabby controls. Samples were pooled according to experiment, processed for the isolation of RNA, and analyzed for differentially expressed mRNAs by using primarily CodeLink Bioarrays, GEM 1 and 2 gene chips and quantitative real-time PCR (qPCR) procedures. Gene chip data were analyzed with GeneSifter.Net software. Our results demonstrate that testosterone regulates the expression of over 2000 genes in the lacrimal gland. Gene ontologies most affected by androgen treatment included those related to cell growth, proliferation and metabolism, cell communication and transport, nucleic acid binding, signal transduction and receptor activities. Our findings also indicate that androgen action may be mediated, at least in part, through classical androgen receptors, and may contribute to the sex-related differences in gene expression of lacrimal tissue. Overall, these results support our working hypothesis that androgen action on the lacrimal gland is mediated primarily through a receptor-associated regulation of gene transcription.

Prospective assessment of proliferative diabetic retinopathy with observations of posterior vitreous detachment.

PURPOSE: To study the relation between posterior vitreous detachment (PVD) and progression of diabetic retinopathy (DR), based on our observation that proliferative DR is rare in patients with complete PVD. METHODS: The medical records of 403 patients with diabetes were reviewed for the relation between progressive DR and the status of PVD and HbA(1c) over 3 years. PVD was classified into none, complete PVD with collapse, complete PVD without collapse, partial PVD with a thickened posterior vitreous cortex, and partial PVD without a thickened posterior vitreous cortex. DR was classified into none, simple, preproliferative, or proliferative. When it became more extensive or when laser treatment or vitreous surgery was performed, the DR was considered progressive. RESULTS: Progression of DR over 3 years occurred in 128/292 (43.8%) eyes with no PVD, 0/14 (0%) eyes with complete PVD with collapse, 2/8 (25%) eyes with complete PVD without collapse, 15/15 (100%) eyes with partial PVD with a thickened posterior vitreous cortex, and 19/74 (25.7%) eyes with partial PVD without a thickened posterior vitreous cortex. Progression of DR occurred significantly more frequently in eyes with partial PVD with a thickened posterior vitreous cortex compared to eyes with complete PVD with collapse (p<0.0001). HbA(1c), did not differ significantly between these two groups (6.9 +/- 0.9% and 7.5 +/- 0.9%, respectively; p = 0.14), although HbA(1c) was significantly higher (p = 0.04) in patients with progressive DR (78 +/- 1.8%) than in patients without progressive DR (7.5 +/- 1.5%). CONCLUSION: Complete PVD is a strong negative risk factor for DR. The PVD status in patients with diabetes should be evaluated.

Are women with Sjögren's syndrome androgen-deficient?

OBJECTIVE: We hypothesize that androgen deficiency is a critical etiologic factor in the pathogenesis of aqueous-deficient and evaporative dry eye in Sjögren's syndrome (SS). We investigated whether women with SS have a deficiency in total androgens. We also examined whether these patients have elevated serum concentrations of estrogens. METHODS: Blood was drawn from women with primary and secondary SS and age matched controls, and analyzed for steroid concentrations by gas and liquid chromatography-mass spectrometry. RESULTS: Our results show that women with SS are androgen-deficient. Concentrations of 5-androstene-3beta,17beta-diol (5-diol), dehydroepiandrosterone (DHEA), dihydrotestosterone (DHT), androsterone-glucuronide (ADT-G), and androstane-3a,17beta-diol-G (3alpha-diol-G) were all significantly reduced in SS sera relative to controls. In contrast, SS was not associated with significant alterations in the serum concentrations of testosterone, androstenedione, estrone, or 17beta-estradiol. These overall findings could not be attributed to the use of oral contraceptives or hormone replacement therapy, because the concentrations of 5-diol, DHEA, DHT, ADT-G and 3a-diol-G were also decreased in patients with SS compared to levels in control women who were not taking exogenous estrogens. CONCLUSION: Our results show that women with SS are androgen-deficient.

The effects of proinflammatory cytokines on cytokine-chemokine gene expression profiles in the human corneal endothelium.

PURPOSE: To determine the effects of proinflammatory cytokines on differential gene expression profiles in the human corneal endothelium (HCE), by using a cDNA expression array. METHODS: A human cDNA expression array technology was used to study the simultaneous expression of HCE incubated with interleukin (IL)-1alpha and tumor necrosis factor-(TNF)-alpha. Gene-specific semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were performed to examine the gene and protein expression patterns revealed by the cDNA expression array, in the presence and absence of proinflammatory cytokines. Moreover, the expression of these genes was studied in ex vivo HCE of donor cornea by RT-PCR. RESULTS: IL-1alpha and TNF-alpha upregulated the expression of 46 of 268 genes for cytokines, chemokines, and their receptors in stimulated HCE. The most upregulated genes in the cDNA expression array, those of monocyte chemotactic protein (MCP)-1 (CCL2), IL-8 (CXCL8), IL-6, and growth-related beta (GRObeta, CXCL2), were studied. Semiquantitative RT-PCR and ELISA analyses revealed the proinflammatory cytokine-mediated changes in the respective gene transcription and protein expression levels. The mRNAs were detected in ex vivo HCE of donor cornea stimulated with proinflammatory cytokines. CONCLUSIONS: HCE can abundantly express cytokines and chemokines through the stimulation of proinflammatory cytokines. The detected genes, those of CCL2, CXCL8, IL-6, and CXCL2, in HCE could facilitate understanding of the inflammatory responses, including the production of keratic precipitates and the correlation between CE and an inflamed cornea or aqueous humor.

Androgen deficiency, Meibomian gland dysfunction, and evaporative dry eye.

OBJECTIVE: We have recently discovered that women with primary and secondary Sjögren's syndrome are androgen-deficient. We hypothesize that this hormone insufficiency contributes to the meibomian gland dysfunction, tear film instability, and evaporative dry eye that are characteristic of this autoimmune disorder. If our hypothesis is correct, we predict: (1) that androgens regulate meibomian gland function, control the quality and/or quantity of lipids produced by this tissue, and promote the formation of the tear film's lipid layer; and (2) that androgen deficiency, due to an attenuation in androgen synthesis (e.g., during Sjögren's syndrome, menopause, aging, complete androgen-insensitivity syndrome [CAIS] and anti-androgen use), will lead to meibomian gland dysfunction and evaporative dry eye. The following studies were designed to test these predictions. METHODS: Experimental procedures included clinical studies, animal models, and histological, biochemical, molecular biological, and biomedical engineering techniques. RESULTS: Our results demonstrate that: (1) androgens regulate the meibomian gland. This tissue contains androgen receptor mRNA, androgen receptor protein within acinar epithelial cell nuclei, and Types 1 and 2 5alpha-reductase mRNAs. Moreover, androgens appear to modulate lipid production and gene expression in mouse and/or rabbit meibomian glands; and (2) androgen deficiency may lead to meibomian gland dysfunction, altered lipid profiles in meibomian gland secretions, tear film instability, and evaporative dry eye. Thus, we have found that anti-androgen therapy in men is associated with meibomian gland disease, a decreased tear film breakup time, and functional dry eye. Furthermore, we have discovered that androgen receptor dysfunction in women with CAIS is associated with meibomian gland changes and a significant increase in the signs and symptoms of dry eye. Of interest, we have also found that androgen deficiency is associated with significant and striking alterations in the neutral and polar lipid patterns of human meibomian gland secretions. CONCLUSIONS: Our findings show that the meibomian gland is an androgen target organ and that androgen deficiency may promote meibomian gland dysfunction and evaporative dry eye. Overall, these results support our hypothesis that androgen deficiency may be an important etiologic factor in the pathogenesis of evaporative dry eye in women with Sjögren's syndrome.