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Severe destruction of the corneal limbus causes conjunctival invasion and subsequent visual loss. Limbal allograft transplantation (LAT) was recently proposed for the treatment of these disorders. However, whether the method functions as a stem cell transplantation of the corneal epithelium remains unclear. This study provided evidence that donor-derived corneal epithelial cells survive long after LAT.Epithelial cells on the paracentral cornea in patients who have undergone LAT were subjected to fluorescence in situ hybridization (FISH) and polymerase chain reaction restriction fragment length polymorphism (RFLP) analysis. X and Y chromosomes were detected using sex chromosome-specific probes in the FISH analysis, and HLA-DPBI antigens were examined in the RFLP analysis. Eyes receiving conventional penetrating keratoplasty (PKP) served as controls.Donor-derived epithelial cells were detected in three of five eyes (60.0%) in the FISH analysis and in seven of nine eyes (77.8%) in the RFLP analysis. Among these eyes, one and three eyes in the FISH and RFLP analysis, respectively, had both donor- and recipient-derived cells. In control PKP eyes, none of the eyes in the FISH analysis and one of eight eyes (12.5%) in the RFLP analysis had donor-derived cells.These results suggest that donor-derived cells survive much longer after LAT than those after PKP, and that LAT may function as stem cell transplantation of the corneal epithelium.

To characterize the type of reactive oxygen species (ROS) produced by excimer photoablation of aqueous solutions and to show the effects of ROS and antioxidants on corneal stromal cells in vitro.Electron spin-resonance spectroscopy was performed using the spin-trapping agent 5,5-dimethyl-1-pyrroline N-oxide (DMPO) for the detection of the superoxide anion and the hydroxyl radical in an acellular DMPO solution irradiated with the excimer laser. Hydroxyl radicals were produced by the Fenton reaction in vitro by the mixture of hydrogen peroxide and ferrous iron (Fe2+), and the effects on cultured corneal fibroblasts were observed by fluorescent microscopy using the cell death marker, propidium iodide (PI) and TdT-mediated dUTP nick-end labeling (TUNEL).Excimer photoablation of a 1% DMPO solution produced a species-specific spin-trapping adduct for the hydroxyl radical ('OH), but not for the superoxide anion or other unidentified free radical. The signals were inhibited dose dependently by the hydroxyl radical scavenger dimethylsulfoxide (DMSO) and an L-ascorbic acid analogue, EPCK-1. The production of *OH in the supernatant of cultured rabbit corneal fibroblasts by the Fenton reaction caused an increase in PI (+) and TUNEL (+) cells by 90 minutes, which was significantly inhibited by the addition of DMSO.Hydroxyl radicals may be partly responsible for stromal fibroblast cell apoptosis after excimer photoablation.

Although IL-10 has been implicated in the pathogenesis of several autoimmune diseases, the mechanisms by which this cytokine mediates inflammatory lesions remain to be elucidated. Exocrine gland destruction is an important early step in the development of Sjögren's syndrome. To better understand the role of IL-10 in Sjögren's syndrome, we made transgenic mice in which the mouse IL-10 gene was regulated by the human salivary amylase promoter. Transgenic expression of IL-10 induced apoptosis of glandular tissue destruction and lymphocyte infiltration consisting primarily of Fas-ligand (FasL)+ CD4+ T cells, as well as in vitro up-regulation of FasL expression on T cells. These data suggest that overexpression of IL-10 in the glands and their subsequent Fas/FasL-mediated bystander tissue destruction is a causal factor in the development of this disease.

To study the cytomorphologic features of bar-shaped chromatin in conjunctival samples and assess whether there are morphologic similarities between nuclear grooves seen in thyroid papillary carcinoma and bar-shaped chromatin.A total of 10 conjunctival samples from five volunteers were studied cytologically. Of the 10 samples, 2 showed barshaped chromatin. This chromatin was studied using a light microscope, scanning electron microscope (SEM) and transmission electron microscope (TEM).On SEM, bar-shaped chromatin was observed as a fissure or shallow cracks. On TEM the barshaped chromatin existed both in the center of the nuclei and in the nuclear membrane and cytoplasm. It was surrounded by outer and inner membranes.It seems that bar-shaped chromatin is formed by the nuclear membrane and that these changes are essentially the same configuration as cytoplasmic invaginations commonly present in papillary carcinoma of the thyroid.

Changes in the ocular surface of patients with Sjögren syndrome (SS) often are more severe than those in patients with dry eye without SS. This study was conducted to investigate the possible involvement of meibomian gland dysfunction in SS-related ocular surface abnormalities.A nonrandomized, prospective, clinical study.Twenty-seven eyes of 27 consecutive patients with SS (SS group) were studied. Twenty-nine eyes of age- and gender-matched non-SS patients with aqueous tear deficiency (non-SS group) were examined as control subjects.Changes in the ocular surface, tear function, and meibomian gland were examined.Tear evaporation rate, meibomian gland expression, and meibography were measured.Fluorescein and rose bengal staining scores were significantly higher in the SS group than in the non-SS group (P = 0.0001). Evaporation of tears was increased significantly in the SS group compared with the non-SS group. There were no significant differences in the rate of tear production between the SS and non-SS groups. Meibography showed that 11 (57.9%) of 19 eyes in the SS group had gland dropout (i.e., histologic destruction of meibomian glands) in more than half of the tarsus. The incidence was significantly higher than that in the non-SS group (5 [18.5%] of 27 eyes; P = 0.005).The results of this study indicate that destruction of meibomian glands and an increase in tear evaporation often are associated with changes in the ocular surface in patients with SS. Severe ocular surface changes in patients with SS may be attributed, in part, to the meibomian gland dysfunction.

A new animal model, the NFS/sld mutant mouse, was used for primary Sjögren's syndrome to investigate the efficacy of topical and systemic cyclosporin A (CyA) in preventing inflammation of the exocrine glands.Cyclosporin A was applied topically (0.01% and 0.1%, three times a day) or administered orally (10 mg/kg and 100 mg/kg, once a day) to mice from 6 to 16 weeks of age, after which the mice were killed.Topical CyA reduced lacrimal gland and submandibular gland inflammation without causing pathologic changes in other organs. Flow cytometry showed that CD44 expression of CD4 T cells from the submandibular lymph nodes was downregulated, whereas that of Mel14+ was upregulated. Using a reverse transcription-polymerase chain reaction assay, we determined that topical CyA significantly decreased the expression of mRNA of IL-2 and T-cell receptor-constant beta-chain.Topical CyA may be clinically useful in reducing the lymphocyte infiltration of lacrimal glands associated with Sjögren's syndrome.

To investigate whether reoxygenation after extended hypoxia causes cellular damage in cultured corneal epithelial cells and to demonstrate the protective effects of lactoferrin.Immortalized human corneal epithelial cells (T-HCECs) were cultured to confluence in 96-well culture plates, subjected to stringent hypoxia (1% O2, 5% CO2, 94% N2 at 37 degrees C) for 24 hours, and returned to normoxic conditions (5% CO2, 95% air at 37 degrees C). Cell viability was observed by 1 microM propidium iodide staining 0, 2, 4, and 6 hours after reoxygenation. Inhibition studies were performed after 2 hours' reoxygenation, using 2 mM iron chelator desferrioxamine and 0.2 mg/ml lactoferrin. Confocal immunocytochemistry for human lactoferrin and western blot analysis for lactoferrin-induced ferritin were performed in cultured T-HCECs to demonstrate the internalization of lactoferrin after application.After 2 hours, reoxygenation of T-HCECs after hypoxia produced an increase in cell death that was significantly greater than that observed in normoxic control cells or in cells subjected to hypoxia for the same time span without reoxygenation. The addition of desferrioxamine and lactoferrin at the time of reoxygenation significantly attenuated cellular damage. Confocal immunocytochemistry revealed that lactoferrin is taken into the cytoplasm of T-HCECs as early as 30 minutes after application. This was also demonstrated in western blot analysis by the upregulation of intracellular ferritin at 18 hours by the addition of iron-bound lactoferrin but not by iron-free lactoferrin.Reoxygenation is responsible for increased cellular damage after extensive hypoxia, which is attenuated by chelators of free iron in the cytosol, including the major tear protein lactoferrin.

Recent clinical trials have indicated that an automated smear apparatus (ThinPrep process) of sample preparation has great diagnostic sensitivity. In this study, conjunctival brush cytology prepared using the ThinPrep method was applied in ocular surface disorders especially for dry eye status. To assess its diagnostic value in cellular samples, 35 patients with keratoconjunctivitis sicca (KCS) and 12 normal volunteer patients were examined using this technique. Conjunctival cells from normal controls revealed fine chromatin and polyhedral cytoplasm without keratinized cytoplasm. However, cellular samples from KCS revealed increased keratinized cells with pyknotic nuclei. They also contained extremely elongated cells. In KCS patients, the mean numbers of keratinized cells were significantly higher (32.3 cells/300 cells) than those of the normal control group (0.2 cells/300 cells). In patients with KCS, inflammatory cell counts were also higher than those of normal controls. Conjunctival cytology by the ThinPrep method obviously deserves additional trials as an adjunct to cytological evaluation of dry eye status, especially for quantitative ocular evaluation of various ocular lesions.

To describe a patient with Terrien's marginal degeneration associated with a chronic skin disease, erythema elevatum diutinum.A 27-year-old woman with peripheral thinning of her left cornea associated with adjacent conjunctival and episcleral inflammation was clinically examined.Episodes of worsening skin lesions were associated with development of inflammation in her left eye. Administration of sulfones was effective for reducing both skin and ocular inflammation.Erythema elevatum diutinum should be considered an underlying systemic disease of Terrien's marginal degeneration.

The adaptive optics system design for the W. M. Keck Telescope incorporates over twenty tracking and movable stages on the optical bench. This paper presents a commercial solution for controlling and positioning these stages. It describes the hardware system and the EPICS software interface used to communicate with the off the shelf hardware controllers. It touches on the positional accuracy and repeatability requirements and the selection of hardware to meet those requirements. It examines the cost and packaging issues and tradeoffs between developing custom hardware and software versus commercially available equipment. A method of synchronizing the stages to absolute time for telescope tracking is also presented.

To evaluate the effects of a lipophilic analog of superoxide dismutase (SOD) in the prevention of polymorphonuclear leukocyte (PMN)-induced damage to corneal epithelial cells in vitro and in bacterial corneal ulcers in vivo.Immortalized human corneal epithelial cells (T-HCEC) were cocultured with human PMNs activated with N-formyl-methionyl-leucyl-phenylalanine for 18 hours, after which lactate dehydrogenase (LDH) activity of the supernatant was measured as a marker of cellular damage. The inhibitory effects of lecithin-bound SOD (PC-SOD) and unmodified SOD, as well as PMNs pretreated with anti-CD 18 monoclonal antibody, were compared with untreated control. The retention of each drug on the ocular surface of healthy volunteers was measured by flow cytometry using brush cytology samples. The protective effects of a 0.1% solution of PC-SOD on Pseudomonas aeruginosa corneal infection in guinea pigs were assessed by inflammatory grading scores and histology.Both PC-SOD and SOD effectively suppressed PMN-induced LDH release in T-HCEC in a dose-dependent manner. LDH release was also attenuated when PMNs were pretreated with anti-CD 18 antibodies, suggesting that adhesion molecules were involved in the process. Brush cytology of conjunctival samples showed that PC-SOD was retained longer on the ocular surface compared with unmodified SOD. PC-SOD significantly prevented excessive tissue damage by infiltrating PMNs in P. aeruginosa corneal infection, whereas in control eyes, perforation of the cornea occurred by 6 days.PC-SOD was effective in attenuating PMN-related tissue damage to corneal tissue both in vitro and in P. aeruginosa infection in guinea pigs.

Previous studies have shown that removal of running sutures after penetrating keratoplasty causes unpredictable changes in astigmatism. The current study was conducted to investigate whether computer-assisted videokeratography is beneficial for predicting visual outcomes after running sutures are removed.The design was that of a prospective clinical study.The authors prospectively studied 29 consecutive eyes undergoing a 10-0 nylon running suture removal after penetrating keratoplasty.Videokeratography was performed before, 1 week, 1 month, and 3 months after removal of sutures.Changes in refractive and topographic astigmatism after suture removal were measured. Topographic patterns and their quantitative descriptors also were analyzed.An asymmetric bowtie was the most common videokeratography pattern both before and after suture removal. After suture removal, the incidence of peripheral corneal steepening increased significantly (2 vs. 21 eyes, P0.0001), and that of focal flattening of the midperipheral cornea decreased (13 vs. 5 eyes, P = 0.046). The mean topographic astigmatism, surface regularity index, and corrected visual acuity were improved significantly by suture removal in eyes that had localized flattening but not in eyes without this finding. Eyes having either skewed axis in astigmatism or topographic astigmatism of more than 9 diopters also showed significant decreases in astigmatism.Suture removal after keratoplasty is advantageous for both reducing astigmatism and normalizing topography, especially in eyes that have localized flattening of the midperipheral cornea. Predictability of visual outcomes of a running suture removal in postkeratoplasty eyes may be improved by the use of videokeratography.

To demonstrate that leukocyte adhesion to cultured corneal endothelial cells is mediated by the CD18 antigen, and to determine whether dexamethasone directly suppresses adhesion by inhibiting activation of nuclear factor kappa B (NFkappaB).Cultured bovine corneal endothelium was stimulated for 6 hours by 40 micron/ml tumor necrosis factor alpha (TNFalpha). Dexamethasone was added 1 hour before TNFalpha stimulation in the dexamethasone group. After stimulation, neutrophils separated from a healthy human volunteer were added with or without anti-CD18 antibody. The culture plate was settled for 15 minutes at 37 degrees C, and then neutrophils were activated by N-formyl-methionyl-leucyl-phenylalanine for 5 minutes. Nonadherent neutrophils were removed by sealing and inverting the culture well. The intracellular localization of NFkappaB after TNFalpha simulation was determined by confocal immunocytochemistry using an anti-p65 antibody.Neutrophil adhesion to cultured corneal endothelial cells increased significantly on exposure to TNFalpha (451.4+/-45.4 cells/mm2, n = 16) compared to control (156.7+/-27.3 cells/mm2, n = 16, P0.01). This increased adhesion was suppressed by the addition of anti-CD18 antibody (157.6+/-25.1 cells/mm2, n = 8, P0.01) and by pretreatment with 10(-7) M dexamethasone (207.9+/-31.5 cells/mm2, n = 10, P0.01). Immunocytochemistry 60 minutes after stimulation revealed that NFkappaB was located in the cytoplasm in unstimulated cells; however, the addition of TNFalpha caused NFkappaB to translocate into the nucleus. Pretreatment with dexamethasone tapered NFkappaB translocation into the nucleus.Leukocyte adhesion to the corneal endothelium was shown to be mediated by CD18 expressed on activated leukocytes. Pretreatment of the endothelium with dexamethasone inhibited leukocyte adhesion; this may be due in part to the suppression of NFkappaB entry into the nucleus.

Recent studies have suggested that the nervous system may participate in inflammatory processes. Substance P (SP) acts as a chemical mediator as well as a neurotransmitter.In order to clarify the pathogenesis of ocular allergic diseases, we assessed the concentration of SP in tears.Using a highly sensitive and specific double-antibody enzyme immunoassay (EIA), we determined the SP concentration in tears of 10 patients with seasonal allergic conjunctivitis, 10 with atopic dermatitis without keratoconjunctivitis (AD), 13 with vernal keratoconjunctivitis (VKC) and 65 normal controls. Giemsa's staining for brush cytology samples and histocytological study by immunocytochemical staining of giant papillary conjunctival cells from VKC and normal controls was conducted.The mean SP level was low in the normal controls and AD, whereas patients with seasonal allergic conjunctivitis and VKC showed significant elevation of SP (P0.01). Brush cytology samples showed conjunctival epithelial cells with lymphocytes, neutrophils and eosinophils that were not seen in normal subjects. Histocytological examination demonstrated SP positive cells in the conjunctiva of patients with VKC, but not in normal controls.This study suggests that the increased level of SP in tears may contribute to the pathogenesis and severity of ocular allergic diseases.

To evaluate the effects of ultraviolet B light (UV-B) on mitochondrial inner membrane function, cell viability, and migration of cultured human corneal epithelial cells.After UV-B exposure in SV-40 transfected human corneal epithelial cells (T-HCEC), mitochondrial function was assessed by digital microfluorography using the mitochondrial marker, rhodamine 123 (Rh 123). The oxygen consumption rate of T-HCEC suspensions (10(7) cells/ml) was measured by an O2 meter, and adenosine triphosphate contents were measured by luciferase-driven chemiluminescence. Cell viability and migration was observed by propidium iodide (PI) staining and migration assays.UV-B exposure caused an immediate drop in O2 consumption by T-HCEC suspensions, whereas exposure of a monolayer culture of T-HCEC to UV-B at radiant exposures of 50 mJ/cm2 caused a reversible decrease in Rh 123 fluorescence (22.4%) and a significant decrease in adenosine triphosphate contents (1.52 +/- 0.05 nmol/10(6) cells) compared to control (2.93 +/- 0.12 nm/10(6) cells) after 10 minutes. The effects on Rh 123 fluorescence were irreversible at 100 mJ/cm2, which approximately corresponded with the threshold dose at which cells positive to PI staining (PI+) appeared. UV-B doses of 50 mJ/cm2 caused detachment of T-HCEC, primarily PI-, whereas higher doses (100 mJ/cm2) resulted in PI+ cells that did not detach from the dish. These PI+ cells hindered the migration of surrounding viable cells; detachment of PI- cells allowed cells to migrate and to cover a trough created by a 500 microns wide beam of UV-B.Threshold levels of UV-B (100 mJ/cm2) are associated with irreversible mitochondrial dysfunction and with the loss of the ability for cultured corneal epithelial cells to detach in vitro.

By exposing a GaAs surface to Cs and O2, a surface which is called negative electron affinity (NEA) surface can be obtained where the vacuum level shifts lower than the bottom of GaAs conduction band. The Cs/GaAs NEA surface has been attracting great attention in recent years, because of its availability for a highly functional electron source which can be controlled by excitation light. In this work, XAFS and X-ray fluorescence measurements were conducted for Cs/GaAs NEA surface prepared on GaAs (001) and (110) substrates. As the result, As and O were considered as near neighbor atoms of center Cs for both of the samples. Most of Cs formed some kind of Cs-oxide and the Cs atoms at interface between Cs-oxide and GaAs were observed as forming Cs-As bond. Taking account of the ratios of coordination numbers and the fluorescence X-ray yield, the Cs-oxide on GaAs (001) surface forms island like structure in which the body/interface ratio is large, and the Cs-oxide on GaAs (110) surface forms layer like structure in which the body/interface ratio is small.

The female subject, 64 years old, subjectively noticed a mild dryness of the eyes and bilateral, submandibular swelling in 1988. The clinical symptom was mild, and such autoantibody and hyper-gammaglobulinemia as often observed in Sjögren's syndrome were not recognized. However, by judging from the results of gum test (9 mL/10 min), salivary gland scan (Grade 2) and the labial biopsy, we diagnosed the case as Sjögren's syndrome. We then administered prednisolone at 40 mg at the onset of treatment and gradually decreasing the dosage over 3.5 years, and the symptoms improved. As an eyelid tumor in the left eye was noticed in 1991 and began to enlarge, the subject was hospitalized in June of 1992. The biopsy of the tumor (2.0 x 1.7 cm) showed marked polyclonal lymphoproliferation with lymphoid follicles which was determined by immunohistological staining. The case was a patient with a lymphoproliferative lesion from a lacrimal gland which is equal to a so-termed pseudolymphoma. Since there are some reports concerning the transition from pseudolymphoma to malignant lymphoma, this patient should be followed carefully.

To investigate whether reflex tearing measured by the Schirmer test with nasal stimulation is better correlated with lymphocytic infiltration of the lacrimal gland and is thus a more appropriate test for Sjögren's syndrome (SS).272 patients with dry eye (age 55.5 +/- 13.5 years) with regular Schirmer test results10 mm were recruited for the study. They were divided into 2 groups depending on the results of the Schirmer test with nasal stimulation. 24 age and sex matched patients from each group were selected for lacrimal gland biopsy and 10 patients for salivary gland biopsy, and the levels of systemic autoantibodies and degrees of ocular surface staining and lymphocytic infiltration of the lacrimal and salivary glands were compared.Of the 272 patients, 187 (68.8%) had good reflex tearing (GT) and 85 (31.2%) had poor reflex tearing (PT). There were no significant differences in the age or sex of the 2 groups. Lymphocytic infiltration of the lacrimal gland was observed in 7 GT cases and 22 PT cases out of 24 cases. Lymphocytic infiltration of the salivary gland was also higher in the PT (6 of 10) than the GT group (2 of 10).Evaluation of reflex tearing by the Schirmer test with nasal stimulation identifies 2 groups of patients with dry eye. Those with poor reflex tearing were more likely to have autoantibodies and lymphocytic infiltration of the exocrine glands consistent with SS.

To determine the diagnostic value in Sjögren's syndrome (SS) of evaluating the presence of focal adenitis in labial salivary gland (LSG) and lacrimal gland (LG) biopsy specimens.We studied 105 cases with suspected SS and performed biopsies prospectively. The diagnostic criteria of Fox, et al were followed. The Chisholm-Mason grading standard and Greenspan focus score were used for evaluation.Epimyoepithelial islands and severe lymphocyte infiltration with germinal centers were observed only in LG biopsy specimens (5.9 and 3.5%, respectively). The lymphocytic focus scores of LG (2.5 +/- 2.1/4 mm2) were significantly higher than those of LSG specimens (1.3 +/- 1.1/4 mm2) in 23 cases with both biopsy results (p0.05). Six of the 32 LSG biopsies were normal while their LG biopsies were significantly higher. Conversely, 5 of the 32 LG biopsies were aided by corresponding LSG biopsies, since in LG biopsies no glandular tissue was obtained. Evaluation of both specimens for the same patient was significantly more effective in diagnosing SS than using either specimen alone (p0.01).LG biopsy specimens have a more evident histopathology than LSG biopsy specimens. We recommend that both LSG and LG biopsies be performed in patients with suspected SS to reduce false negative results and improve diagnostic accuracy.

Allergic conjunctivitis caused by sugi pollen is considered to be strictly a "spring disease". However, a recent report indicated that sugi pollen is scattered not only in spring but in all seasons, especially in the autumn. We retrospectively determined the number of patients with allergic conjunctivitis caused by sugi pollen during each month for 3 consecutive years, and also investigated the growth of sugi male cones in November for four years. Diagnosis of allergic conjunctivitis was based on symptomatic complaints, clinical findings, and serum sugi pollen specific IgE measured by the Multiple Antigen Simultaneous Test 16. The annual incidence of allergic conjunctivitis peaked twice, once in spring and again in autumn. Scattering of sugi pollen occurred mostly in March and October, whilst the growth of sugi male cones was highest in November. Thus, allergy to sugi pollen can cause allergic conjunctivitis both in the spring and late autumn.

DNA-HLA (human leukocyte antigen) typing was performed for ocular tissues in order to determine the usefulness of the method in corneal transplantation. Each type of ocular tissue was dissected from eye bank eyes (n = 3). DNA was extracted, and HLA-DRB1, DQB1, and DPB1 genes were amplified using PCR (polymerase chain reaction) method. DNA can be extracted and amplified from each ocular tissue except for the crystalline lens, but DNA from iris and choroid can be amplified only after diluting the samples 10 to 100 times. HLA class II antigens were successfully determined in these ocular tissues by the RFLP (restriction fragment length polymorphism) method. The method was applied to the corneal tissues of donor and recipient used for the corneal transplantation (n = 7). HLA class II antigens can be determined in both the donor and recipient corneal samples, and the results of recipients' HLA typing were the same as those determined using the blood samples. These results indicate that the DNA-HLA typing can be used for ocular tissues. Since DNA-HLA typing is more accurate than the conventional serological typing, the method is promising for the study of HLA class II typing in corneal transplantation.

In order to investigate the effect of occlusion on the skin, the flexor sides of both upper arms were covered with column-shaped with column-shaped closed chambers, 30 mm in outer diameter, 20 mm in inner diameter, and 5 mm in height, which were made of polyethylene foam. The tops of the chambers were sealed by plastic films with various levels of water vapor permeability to control moisture in each chamber. The raised chamber walls prevented direct contact between the skin and the plastic film. After 24 h of application, morphological changes of the skin surface were observed microscopically by the nitrocellulose-replica method. Although no visual alterations were found on all areas of occluded skin, microscopic evaluation showed that simple occlusion by films induced an increase in the number of deepened skin furrows on the skin surface. this increase was associated with lower water vapor permeability of the films, as well as with higher values of both temperature and humidity of the test day. Thus, since conditions which facilitate perspiration from the skin tend to cause skin irritation, prolonged exposure of the skin to sweat by simple occlusion may act as a primary skin irritant.

We evaluated the effects of stellate ganglion block (SGB) on conjunctival oxygen tension, ocular surface temperature (n = 7) and intraocular pressure (n = 18) in patients with retinal vein occlusion and other retinopathy. The conjunctival oxygen tension (PcjO2) increased from 45.6 +/- 5.0 mmHg to 76.3 +/- 19.9 mmHg in 45 minutes by SGB and then gradually returned to the control level. PcjO2 120 minutes following SGB remained still at a higher level compared with the control value. Ocular surface temperature increased from 35.1 +/- 0.3 degrees C to 35.9 +/- 0.3 degrees C 25 minutes following SGB. Intraocular pressure decreased below the control level from 15 minutes to 120 minutes by SGB following an initial temporary increase. In conclusion, we have found that SGB does increase oxygen tension in ocular tissue and decrease IOP.

The number of the cells and the layers of the conjunctiva which can be obtained by brush cytology was studied on rabbit conjunctiva using cytological and histological methods. About 1,000, 3,500, and 10,000 conjunctival epithelial cells can be obtained by 7, 14, and 20 brush strokes histological observations showed that all varieties of the layers including basal cells were obtained without destroying the basement membrane. This study showed that the brush cytology can offer single cell suspension only from the conjunctival epithelium, and that brush cytology is useful to investigate the conjunctival epithelium by flow cytometry, polymerase chain reaction, and other methods.

We have reported in the past a car crash in which the air bag system might have caused driver's corneal endothelial cell loss. To evaluate the corneal endothelial cell loss caused by air bag systems, we performed several air bag impact tests. We found that the corneal endothelial cell loss was greater at a distance of 240 mm than other distances. We also found that the corneal endothelial cell loss caused by the air bag which has bigger inflater and heavier material was significantly greater than that caused by other systems. This study offers basic data to improve the safety of air bag systems.

The metabolic pathways of glycolysis and mitochondrial respiration in the corneal endothelial cell are the primary sources of the adenosine triphosphate (ATP) necessary to maintain endothelial structure and pump fluid to maintain the corneal stroma in its normally dehydrated and transparent state. The correlation between endothelial metabolism and morphology in rabbits was studied for 7 days after the application of three different agents: (1) iodoacetamide, used to inhibit ATP synthesis from both glycolysis and respiration; (2) potassium cyanide (KCN), used to inhibit ATP synthesis from respiration only; and (3) ouabain, used to inhibit fluid pumping but not ATP synthesis. After application of each of these three drugs to the corneal endothelium, changes in endothelial morphology were measured. The greatest change resulted from the use of iodoacetamide. Specular microscopic examination of the endothelium after the application of iodoacetamide showed progressive degradation of the integrity of the cellular structure; after 6 hr, there were no discernible cell borders. In those corneas treated with either KCN or ouabain, only minor changes in the endothelium were seen during the full 7 days of the investigation. Computer-assisted morphometric analysis showed an increase in the coefficient of variation of both cell area and perimeter in all cases. This increase was greater in the corneas treated with ouabain than those treated with either iodoacetamide or KCN. Redox fluorometry showed that the metabolic ratio (autofluorescence of reduced pyridine nucleotides divided by that of oxidized flavoproteins) decreased significantly in the iodoacetamide-treated corneas, increased significantly in the KCN group, and showed no significant change in the corneas in the ouabain group, all compared with a control group. The results showed that (1) when ATP produced by both glycolysis and respiration was inhibited by 0.1 mmol/l iodoacetamide, the endothelial cells could not survive, but (2) when ATP synthesis produced by respiration alone was inhibited by 1.0 mmol/l KCN, the cells could survive for at least 1 wk on the ATP produced by anaerobic glycolysis. Furthermore, the polymegathism seen after application of ouabain, a drug that is not believed to affect ATP synthesis but inhibits the endothelial pump function, is greater than that seen as a result of reduced pump function caused by inhibited respiration produced by 1.0 mmol/l KCN. Combining specular microscopy, computer-assisted morphometric analysis, redox fluorometry, and corneal pachymetry allowed correlations between corneal endothelial metabolism, pump function, and morphology to be studied.

A simple method was developed to measure tear evaporation. A sensor was inserted into a chamber covering the eye. The humidity inside each chamber then was measured every 10 sec for 2 min with both eyes either closed or open but allowing normal blinking. The difference between these conditions represented evaporation from the ocular surface. Using this method, the tear evaporation rate at 40% ambient humidity (TEROS 40) was calculated. The average TEROS 40 in normal subjects (n = 43) was 15.6 +/- 3.8 x 10(-7) g/sec. It was 9.5 +/- 5.6 x 10(-7) g/sec in patients with dry eye symptoms (n = 72, P less than 0.001). The insertion of lacrimal collagen implants in one group of such patients (n = 10) increased the TEROS 40 from 10.2 +/- 5.5 x 10(-7) g/sec to 18.2 +/- 4.8 x 10(-7) g/sec (P less than 0.01). The instillation of eye drops increased the TEROS 40 significantly in patients with dry eye symptoms for at least 1 min (n = 10, P less than 0.01); a continued effect depended on the type of eye drop. Increased TEROS 40 still was observed 5 min after instillation of viscous eye drops (0.1% and 0.3% sodium hyaluronic acid); the TEROS 40 returned to original levels within 5 min after instillation of artificial tears of normal viscosity with or without 0.05% sodium hyaluronate. In all cases, TEROS 40 returned to original levels within 10 min. This was a quick reliable method for measuring tear evaporation from the ocular surface, and it can be applied to evaluate tear dynamics and subclassifications of dry eyes.

Oxygen free radical formation by conventional phacoemulsification devices has been postulated as a possible mechanism of corneal endothelial damage during surgery. To test this hypothesis, phacoemulsification probe-induced free radical production was visualized using a single photon-counting camera and an O(2-)-sensitive luciferin derivative, 2-methyl-6-[p-methoxyphenyl-3,7-dihydroimidazo [1,2-a]pyrazin-3-one (MCLA), which allows the visualization of spatial and temporal alterations in free radical production. Within 1 min after starting ultrasound emission, MCLA-dependent chemiluminescence was increased significantly, the intensity of which was maximal at the tip of the probe and tapered along a gradient toward distal portions. The chemiluminescence was suppressed significantly by adding either superoxide dismutase (300 U/ml) or sodium azide (20 mmol/l). By adding deuterium to the medium, MCLA-dependent chemiluminescence significantly increased, suggesting the involvement of singlet oxygen in the reaction.

Today, as with western countries, diagnosis and treatment of Sjögren's syndrome (SS) patients in Japan are carried out by a team effort. The clinical manifestations of Japanese SS patients are quite similar also. There are, however, several minor modifications in the diagnosis and treatment of SS patients in Japan, which are presented in this article.

Redox states of the corneal endothelium in 42 recipient corneas obtained at the penetrating keratoplasty were measured non-invasively using ocular redox fluorometry. Autofluorescence from reduced pyridine nucleotides (PN) and oxidized flavoproteins (Fp) were measured, and the PN/Fp ratio was used as an indicator of the redox state. Endothelial damage was graded as normal, mildly damaged, moderately damaged, and severely damaged, based on the histopathological findings. Mildly damaged endothelium showed a significantly higher PN/Fp ratio than the that in normal endothelium whereas the ratio was significantly lower in the severely damaged endothelium. These changes in the redox state may represent compensation and decompensation processes of the endothelial metabolism. Ocular redox fluorometry was shown to be useful for the evaluation of the metabolic state in the human corneal endothelium.

Abnormalities in glucose metabolism are thought to be among the main causes of cataract formation. We have taken noninvasive biochemical measurements of the lens which provides us with information concerning glucose metabolism in the lens epithelium. The autofluorescence of reduced pyridine nucleotides (PN) and oxidized flavoproteins (Fp) in the rat lens epithelium was measured noninvasively as a function of time using redox fluorometry. The oscillations of the metabolic ratio, PN/Fp, were measured in vivo, in situ, and in the organ-cultured lens. The PN/Fp ratio in the organ-cultured lens ranged from 1.05 to 2.57 within a period of 60-90 minutes (mean +/- SD = 1.52 +/- 0.36). This PN/Fp ratio increased by 23% when a respiratory inhibitor (8 mM KCN) was applied to the lens. However, it decreased by 10% in the presence of a complete metabolic inhibitor (8 mM iodoacetamide). The presence of metabolic oscillations in the in vivo, in situ and cultured lens indicates that this oscillation is a local phenomenon. In cell-free extract systems, oscillations of several intermediates in the glycolytic pathway have been previously demonstrated and this PN/Fp oscillation is thought to be a reflection of glycolytic oscillation.

A new method for measuring tear evaporation from the ocular surface is developed. The change of humidity in the closed chamber around the eye was recorded, with eyes open and eyes closed. Theoretical equations were used for data analysis, and the evaporation coefficient K, the tear evaporation volume at the environmental humidity of 40% were calculated. We examined 28 normal eyes and 58 eyes of dry eye patients. Evaporation coefficient K (x 10(-4)/sec) were 32.4 +/- 9.2 in normal and 16.5 +/- 11.3 in dry eyes (p less than 0.001). Tear evaporation volume at the environmental humidity of 40% (x 10(-7) g/sec.eye) were 16.2 +/- 4.7 in normal and 8.4 +/- 5.7 in dry eyes (p less than 0.001). This method is so simple and non-invasive that it could be used as a new examination for dry eye.

A special brush was developed for collection of conjunctival cells with better integrity of the harvested specimens. Using this brush, cells were collected from temporal bulbar conjunctiva of 20 normal eyes and 40 eyes with dry-eye, and were stained by the modified Papanicolau method, Keratinized cells (K) and goblet cells (G) were classified into 5 categories, from (-) to ( ) according to the numbers of the cells observed. Normal eyes were categorized as K (-):16, K(+/-):4, G (+):2, G(++):18, Eyes with dry-eye were K (-):3, K (+/-):8, K (+) 13, K (++):11, K ( ):5, and G (-):7, G (+/-):7, G (+):22, G (++):4. Clinical severity and the rose Bengal score corresponded to these categories. This technique can detect cytological changes in conjunctival cells in terms of shape and quality of staining. Eyes with dry-eye could be evaluated in relation to the numbers of keratinized cells and goblet cells.

Japan is situated at the opposite end of the European continent from Western Europe. Climate, culture and history all differ greatly between the two, as does agriculture. Half of the farmland is devoted to paddy rice, livestock raising is still a new industry and the average farm size is only one hectare. Nevertheless, one would find many similarities in policy. The oversupply of major agricultural products is one of the most serious problems in Japan, whilst slow structural reform is the other, and a declining rural community is undermining the agricultural base. This paper confines itself to an outline of Japanese agricultural policy, without making explicit comparisons with Western Europe.

The biochemistry of contact lens-cornea interaction is not well understood, although previous studies have suggested that corneal metabolic changes may be the underlying factor in morphological alterations. Using a rabbit model, this interaction has been examined with 31P nuclear magnetic resonance (NMR) spectroscopy, which detects signals principally from the epithelium. The examination was supplemented with electron microscopy and histochemistry. Polymethylmethacrylate lenses caused reversible changes, including activation of anaerobic glycolysis and disturbance of membrane metabolite levels. These changes were far more severe than those occurring during prolonged eye closure. There appears to be an association between cellular deterioration and loss of membrane metabolites. On the other hand, oxygen-permeable silicone lenses allowed maintenance of nearly normal metabolic patterns. These results show multifaceted corneal response to hard contact lens wear.