Sensitivity and specificity of photopic negative response of focal electoretinogram to detect glaucomatous eyes.

AIMS: To determine the sensitivity and specificity of the photopic negative response (PhNR) of the focal electroretinograms (ERG; focal PhNR) to detect glaucomatous eyes with different degrees of visual field defects. METHODS: One-hundred and fourteen eyes of 114 patients with open angle glaucoma and 42 eyes of 42 normal controls were studied. The focal ERGs were elicited by a 15 degrees stimulus spot centred on the macula, and on the supero-temporal and on the infero-temporal areas of the macula. The receiver operating characteristic curves were determined to obtain optimal cut-off values. Eyes were classified as being glaucomatous when their focal PhNRs were less than the cut-off values in either retinal area (combined criterion). RESULTS: The focal PhNR amplitudes were significantly reduced with an advance in the stage of glaucoma. In early glaucoma, the sensitivities of the PhNR measured for each retinal area ranged from 58.1% to 80.7%. The sensitivities were significantly increased to 90.6% and 96.9% for the focal PhNR amplitude and the focal PhNR/b-wave amplitude ratio, respectively, when the combined criterion was employed. The specificity was >90%. CONCLUSIONS: Focal PhNRs have diagnostic ability in detecting early glaucoma with high sensitivity and specificity, especially when the combined criterion is used.

The effect of endostatin evaluated in an experimental animal model of collagen-induced arthritis.

OBJECTIVE: To investigate the arthritis-inhibiting effect of endostatin, a potent angiogenesis inhibitor, on type II collagen-induced arthritis (CIA). METHODS: In an experimental system of prophylactic administration, endostatin was administered once daily at 1 mg/kg/day or 10 mg/kg/day for 2 weeks from before the onset of arthritis. In the experimental system of therapeutic administration, mice with an arthritis score of 1 to 3 were administered endostatin once daily at 10 mg/kg. In the experimental system of continuous administration, endostatin was administered using an osmotic pump capable of continuously administering a calculated dose of 1 mg/kg/day for 2 weeks. RESULTS: Arthritis scores were lower in a dose-dependent manner in the prophylactic administration group than in the control group. Arthritis scores were lower in the therapeutic administration group than in the control group. Compared with the once-daily dosage regimen, the administration of endostatin by an osmotic pump achieved a similar arthritis-inhibiting effect at one-tenth of the dose. CONCLUSION: Both prophylactic and therapeutic administration of endostatin inhibited type II CIA in mice. The administration method using an osmotic pump is useful.

Inhibition of arthritis by systemic administration of endostatin in passive murine collagen induced arthritis.

OBJECTIVE: To investigate the arthritis inhibiting effect of endostatin, known to have potent antiangiogenic activity, systemically given to animal models of rheumatoid arthritis (RA). METHODS: Four kinds of monoclonal anti-type II collagen antibody followed by lipopolysaccharide (LPS) three days later were given to 6 week old, female Balb/c mice to induce arthritis. Three groups of mice received 0.2 mg/kg/day, 2 mg/kg/day, and 10 mg/kg/day of endostatin, respectively, whereas a control group received phosphate buffered saline (PBS). Endostatin or PBS was given for 13 days, starting before the development of arthritis. Arthritis was evaluated by arthritis scores and hind paw thicknesses. Mice were killed for histological examination on the 22nd day after the administration of monoclonal anti-type II collagen antibody. RESULTS: Arthritis developed within three days after LPS administration in both the control and endostatin treatment groups. No difference in the development rate of arthritis was noted between the control and endostatin treatment groups. Arthritis scores remained significantly lower in the endostatin 10 mg/kg/day group than in the control group. Hind paw thicknesses also remained significantly smaller in the endostatin 10 mg/kg/day group than in the control group. Histopathological examination showed that synovial thickening and subchondral bone erosion improved more in the endostatin treatment groups than in the control group. CONCLUSION: The systemic administration of endostatin had an arthritis inhibiting effect in RA animal models. Endostatin inhibited, in particular, pannus formation and bone destruction.

Membranous proliferation of lens epithelial cells on acrylic, silicone, and poly(methyl methacrylate) lenses.

PURPOSE: To determine whether intraocular lens (IOL) material influences the membranous proliferation of lens epithelial cells (LECs) on the anterior surface of the IOL. SETTING: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan. METHODS: This prospective study included 87 eyes of 87 consecutive patients who had cataract surgery with IOL implantation. Patients were randomly assigned to receive a single-piece poly(methyl methacrylate) (PMMA) IOL, a 3-piece silicone IOL, or a 3-piece acrylic IOL. Postoperatively, the IOL optic was examined by slitlamp microscopy at 1 and 10 days and 1, 2, and 3 months to determine the incidence, duration, and extent of membranous LEC proliferation. The optic was also examined at 6, 9, and 12 months if proliferation persisted. RESULTS: Thirteen eyes were excluded from analysis because of incomplete coverage of the IOL rim by the anterior capsule margin; 74 remaining eyes were studied. All patients obtained good postoperative visual acuity (20/25 or better). Incidence (P =.0024) and duration (P =.0002) of membranous LEC proliferation with the acrylic IOL was greater than with the other 2 types. Proliferation on the acrylic IOL was more extensive than on the PMMA IOL 10 days postoperatively (P <.05) and on the silicone IOL at 10 days (P <.01) and 1 month (P <.01). However, LEC proliferation on the 3 types of IOLs usually decreased by the third postoperative month, and proliferation did not disturb visual acuity. CONCLUSIONS: The material of an IOL influenced membranous proliferation of LECs on its anterior surface. Proliferation on the acrylic IOL persisted longer than on the other 2 types but did not cause visual symptoms.

Teaching continuous curvilinear capsulorhexis using a postmortem pig eye with simulated cataract(2)(2).

We modified an inexpensive, easily prepared model using a postmortem pig eye for training ophthalmology residents to perform continuous curvilinear capsulorhexis. To reduce the tension and elasticity of the porcine anterior lens capsule, 0.05 mL of formalin mixed with hydroxyethylcellulose or a viscoelastic material is injected into the anterior chamber via the corneal limbus to fix the surface of the central anterior lens capsule of the pig eye in situ. One or 2 minutes after the injection, only the anterior lens capsule is fixed and corneal transparency is maintained. The reduction in tension and elasticity of the anterior lens capsule caused by its fixation increases the resemblance of the simulated cataract to the human cataract.

[Presence of alpha 5 beta 1 integrin and fibronectin in the anterior subcapsular cataract].

PURPOSE: We investigated whether alpha 5 beta 1 integrin and fibronectin were present in myofibroblast-like lens epithelial cells in anterior subcapsular cataract (ASC). METHODS: Nine anterior capsule specimens were obtained from the patients during cataract surgery and frozen for cryostat sections. Six specimens were anterior capsule obtained from cataract with ASC. As a control, three specimens were obtained from cataract without ASC. Alpha-smooth muscle actin (alpha-SMA), alpha 5 beta 1 integrin, and fibronectin were detected by immunohistochemical observation. RESULTS: In all 6 specimens from patients with ASC, the lens epithelial cells around fibrosis tissue included myofibroblast-like lens epithelial cells which were positive for alpha-SMA. alpha 5 beta 1 integrin was detected in these lens epithelial cells. Fibronectin was also detected around these myofibroblast-like lens epithelial cells. Three control specimens showed no immunoreactivity against alpha-SMA, alpha 5 beta 1 integrin, or fibronectin. CONCLUSIONS: Alpha 5 beta 1 integrin and fibronectin may play an important role in myodifferentiation of lens epithelial cells.

Intact corneal epithelium is essential for the prevention of stromal haze after laser assisted in situ keratomileusis.

AIMS: To determine the effect of intact corneal epithelium on stromal haze and myofibroblast cell formation after excimer laser surgery. METHODS: Denuded epithelium alone, photorefractive keratectomy (PRK), laser in situ keratomileusis (LASIK), or LASIK with denuded epithelium was performed in rabbit eyes. Postoperative anterior stromal haze was assessed employing a standard scale. Immunohistochemical methods were used to detect alpha smooth muscle actin (alpha-SMA), a marker for myofibroblastic cells, and type III collagen in subepithelial corneal tissue. RESULTS: Three weeks after surgery, the presence of alpha-SMA positive long extended and spindle-shaped stromal cells, and synthesis of type III collagen were observed in the subepithelial stromal layer corresponding to corneal haze in PRK and LASIK with denuded epithelium, but not in denuded epithelium alone and LASIK. CONCLUSION: The intact corneal epithelium may play an important part curbing subepithelial haze and differentiation of myofibroblasts in corneal wound healing.

Intraocular lens subluxation in a patient with facial atopic dermatitis.

A 66-year-old Japanese man presented with subluxation of a posterior chamber intraocular lens (IOL) caused by a rupture of part of Zinn's zonule but no retinal break 2 years after phacoemulsification with IOL implantation. He had a history of atopic dermatitis since infancy. This case presents a rare ocular complication of scratching and rubbing the face and eyelids because of itching related to atopic dermatitis.

[A case of rheumatoid arthritis associated with autoimmune hemolytic anemia due to weekly low-dose methotrexate therapy].

A 57-year-old woman was found to have rheumatoid arthritis (RA) in 1996. Treatment with different immunoregulatory agents, including actarit and bucillamine, produced no improvement. Therefore, combined therapy with methotrexate (MTX) (5 mg/week oral) and low-dose prednisolone (PSL) (5 mg/day) was started in April 1997. Sulindac and famotidine were also administered. In August 1997, she was admitted to our hospital because of palpitations and shortness of breath due to severe anemia. Results of laboratory studies were hemoglobin, 2.9 g/dl; reticulocyte count, 225/1000; and haptoglobin, less than 10 mg/dl. The direct and indirect Coombs'tests were positive. A diagnosis of autoimmune hemolytic anemia (AIHA) was made on the basis of the laboratory findings. Treatment with high-dose PSL (50 mg/day) was started, and the anemia improved. The hemoglobin level increased to 6.0 g/dl within the 1st week and to 12.6 g/dl 6 weeks later. We believe that the most likely explanation for this anemia was the low-dose MTX because the anemia appeared soon after treatment was started.

[A case of systemic lupus erythematosus associated with superior vena cava syndrome].

We report on a case of systemic lupus erythematosus associated with superior vena cava syndrome. A 46-year-old woman developed polyarthralgia in December 1994. She was treated with nonsteroidal anti-inflammatory drugs. In February 1995, she was admitted to our hospital with systemic convulsion and disturbance of consciousness (III-300/Japan coma scale). Severe facial edema was also present. Laboratory studies revealed the presence of anti-nuclear antibody, anti-DNA antibody, anti-Sm antibody, and proteinuria. An X-ray film of the chest showed pericardial effusion and bilateral pleural effusions. Computed tomography of the chest showed a severe swelling of mediastinal lymph nodes. A diagnosis of systemic lupus erythematosus was made according to the American Rheumatism Association criteria. Initial treatment with intravenous dexamethasone improved the level of consciousness and decreased the facial edema, mediastinal lymphadenopathy, and the effusions on computed tomography of the chest. We believe that the most likely explanation for the facial edema is superior vena cava syndrome due to severe mediastinal lymphadenopathy.

Tranilast inhibits cell proliferation and collagen synthesis by rabbit corneal and Tenon's capsule fibroblasts.

PURPOSE: To determine whether tranilast, N-(3,4-dimethoxycinnamoyl) anthranilic acid, influences cell proliferation and collagen synthesis by rabbit Tenon's capsule fibroblasts (TFs) and corneal stromal fibroblasts (CFs). METHODS: Rabbit TFs and CFs (7000 cells/well) were cultured in F-12 nutrient mixture supplemented with 1% FBS, plus 0, 3, 30, or 300 microM tranilast, and the number of cells was counted 72 hrs later. To determine the effect of tranilast on collagen synthesis, cells at confluence were cultured in a medium containing 0, 3, 30, or 300 microM tranilast and labeled with 3H-proline, and the amount of radioactivity incorporated into collagenase-sensitive proteins was measured. RESULTS: At 300 microM, tranilast decreased the number of TFs by about 27% and the number of CFs by about 45%, but had no effect on cell viability. The same concentration of tranilast reduced TFs collagen synthesis and CFs collagen synthesis. CONCLUSIONS: Tranilast may inhibit scar formation after trabeculectomy for glaucoma and after excimer laser photorefractive keratectomy.

Effect of rabbit aqueous humor obtained after cataract surgery on collagen gel contraction induced by bovine lens epithelial cells.

We evaluated the effect of specimens of pre- and postoperative aqueous humor on the contraction of collagen gels, and the effect of transforming growth factor-beta(2) (TGF-beta(2)) in postoperative aqueous humor. Rabbit aqueous humor was collected preoperatively and on postoperative day 7. Bovine lens epithelial cells (LECs) were cultured in collagen gel in F-12 nutrient mixture supplemented with 5% fetal bovine serum that contained 10% aqueous humor obtained under various conditions. Gel area was determined on day 4. Gels cultured with the medium that contained phosphate-buffered saline showed a statistically significant contraction after 4 days. Aqueous humor from aphakic or pseudophakic eyes significantly increased contraction, with both specimens having a similar effect. Approximately 60% of the contractile effect of the postoperative aqueous humor was neutralized by anti-TGF-beta(2) antibody. However, the promoting effect of the aqueous humor sampled postoperatively was less than that sampled preoperatively. Although the aqueous humor obtained postoperatively increased the contractility of the LECs, with the level of TGF-beta(2) apparently responsible for much of its effect, the effect was less than that observed in the aqueous humor obtained preoperatively.

[Therapeutic efficacy of cyclosporin A in four cases of amyopathic dermatomyositis with rapidly progressive interstitial pneumonia].

OBJECTIVES: We investigated the clinical features of amyopathic dermatomyositis (ADM) with rapidly progressive interstitial pneumonia (IP), and the effectiveness of cyclosporin A (CsA) for IP. METHODS: In our four patients who were accorded with the notion of amyopathic dermatomyositis proposed by Euwer and treated with CsA for IP, and eight reported patients for whom we could obtain details of the clinical course, we investigated the relationship between the clinical features of them at the start of CsA administration and the clinical effectiveness of CsA. RESULTS: When we investigated the effectiveness of CsA in our four patients and eight reported patients, we could judge that effective cases were four and ineffective cases were eight. We compared the PaO2 at the start of CsA administration between effective and ineffective cases, the average PaO2 at the start of CsA administration was 65.7 mmHg in four effective cases, while it was 51.0 mmHg in eight ineffective cases. There was a statistically significant difference between the groups. CONCLUSIONS: Because our results suggested that many ineffective cases were treated with CsA after they had developed severe respiratory failure, we think that CsA should be given to ADM patients with rapidly progressive IP before the progression of respiratory failure.

Extracellular matrixes influence alpha-smooth muscle actin expression in cultured porcine lens epithelial cells.

PURPOSE: To determine whether extracellular matrix (ECM) influences the expression of alpha-smooth muscle actin (alpha-SMA), a marker for myofibroblasts, in cultured porcine lens epithelial cells (LECs). METHODS: Porcine LECs were cultured for 6 days in an F-12 nutrient mixture supplemented with 5% fetal bovine serum on wells that were coated with laminin, fibronectin, type I collagen, or type IV collagen. LECs cultured on uncoated wells served as a control. Alpha-SMA was detected immunocytochemically with a mouse monoclonal antibody, and the ratio of the number of alpha-SMA-positive cells to the total number of cells, the P/T ratio, was calculated. RESULTS: The P/T ratio of the LECs on the uncoated dishes was about 5%. LECs cultured on wells coated with laminin or type IV collagen significantly reduced the ratio, whereas fibronectin or type I collagen had no effect. CONCLUSIONS: The ECM influences alpha-SMA expression in cultured porcine LECs.

Fibrous membrane formation at the capsular margin in capsule contraction syndrome.

PURPOSE: To determine whether the pathogenesis of capsule contraction syndrome involves the outgrowth of the fibrous membrane from the anterior capsule margin. SETTING: Department of Ophthalmology, Keio University School of Medicine, Tokyo, and the Ando Eye Clinic, Kanagawa, Japan. METHODS: A retrospective review of medical records and slitlamp photographs was conducted in 12 eyes (10 patients) that had required treatment for a narrowed anterior capsule opening after cataract surgery. All patients had had continuous curvilinear capsulorhexis and phacoemulsification with implantation of an intraocular lens in the capsular bag. Specimens of surgically removed fibrous membrane were examined by histopathological methods. RESULTS: Fibrous membrane on the inner surface of the anterior capsule and the linear folds of the anterior capsule were present in each eye. In 10 eyes of 8 patients, the fibrous membrane was on the outer surface of the anterior capsule and covered the capsular folds at its margin. Pathological study showed that this fibrous membrane consisted of the flattened lens epithelial cells that proliferated on the inner and outer surfaces of the shrunken anterior capsule. The outgrowth of this membrane from the margin of the anterior capsule to the center of the opening of the anterior capsule was noted. CONCLUSION: In this study, capsule contraction syndrome involved contraction of the fibrous membrane as well as its outgrowth from the capsule margin.

Comparative studies of different stromal cell microenvironments in support of human B-cell development.

This study compared human murine stromal cells for their capacity to support human hematopoietic stem cell (HSC) development into the B lineage. FACS sorted human fetal bone marrow (BM) HSC (CD34+CD19- or CD34+/CD10-/CD19-/CD45RA) were cultured on human fetal BM stromal cells, human skin fibroblasts, or murine S17 stromal cells and analyzed by flow cytometry or reverse transcriptase polymerase chain reaction. CD34+CD19- HSC on human BM stromal cells or fibroblasts differentiated into B-lineage cells with a continuum in density of surface CD19 expression, and some cells expressing micro/kappa or micro/lambda B-cell receptors. In contrast, CD19+ cells from S17 cultures had two- to fourfold higher levels of CD19, but no cells expressing B-cell receptors. The number and percentage of CD19+ cells was high, intermediate, or low in the human BM, human fibroblast, or murine S17 stromal cell cultures, respectively. Reverse transcriptase polymerase chain reaction analysis showed that TdT, CD19, and DHQ52-J(H) rearrangements were expressed at comparable levels when CD34+/CD19- HSC were plated on human or murine stromal cells. In contrast, CD34+/CD10-/CD19-/CD45RA HSC plated on human or murine stromal cells expressed CD19 in both cultures, but TdT was only expressed in human stromal cell cultures. We conclude that human BM stromal cell, human skin fibroblasts, and murine S17 stromal cell cultures can provide complementary and comparative tools for identification of stromal cell ligands with potentially unique functions in regulating human B-cell development.

[Extracellular matrix influencees proliferation of cultured porcine lens epithelial cells].

PURPOSE: To investigate whether extracellular matrix (ECM) influences the proliferation of cultured lens epithelial cells (LECs). MATERIAL AND METHODS: Porcine LECs were cultured in F-12 nutrient mixture supplemented with 5% fetal bovine serum (FBS) for 24 or 96 hours on the dishes coated with laminin, fibronectin, type I collagen, or type IV collagen. As a control, LECs were cultured on uncoated dishes. Twenty-four or ninety-six hours later, the number of cells was determined. We determined the proliferation ratio of the number of cells 96 hours after plating to the number of cells 24 hours after plating. This ratio was used to assess the cell proliferation. RESULTS: The ratio of the LECs on the uncoated dishes was 2.3. Dish coating with type I or type IV collagen, and fibronectin significantly increased this ratio (4.0, 3.5, and 3.0, respectively), whereas coating with laminin did not affect this ratio (2.5). CONCLUSION: These findings suggest that ECM influences cultured porcine LEC proliferation.

Creating cataract in a pig eye.

The purpose of this study was to create a uniform, reproducible hardened lens nucleus and an anterior capsule in a pig eye that approximate those in the human eye for use in cataract surgery training. The lens nuclei was hardened and anterior capsule elasticity was reduced in pig eyes by injecting the lens with mixtures of formalin and alcohol (ethanol, 2-propanol, or both). A 30 gauge needle was used to pierce the lens 3.0 mm from the limbus. Then, 0.2 mL of the formalin and alcohol mixture was injected. The most effective mixture was formalin, ethanol, and 2-propanol in the ratio 4:3:3. Fifteen minutes after this mixture was injected, the pig eye had nuclear cataract with grade III hardness and an anterior capsule that approximated the capsule in a human cataractous eye. The cataract created by this method will be useful in teaching continuous curvilinear capsulorhexis, divide and conquer and phaco chop phacoemulsification, and nuclear hydrodelineation and rotation.