Showing papers in "Current Eye Research in 2000"
TL;DR: Preserved human AM expresses mRNAs for a number of growth factors and contains several growth factor proteins that might benefit epithelialization after AM transplantation, suggesting an epithelial origin for these growth factors.
Abstract: Purpose. To investigate the expression of growth factor mRNA and the level of growth factor protein in preserved human amniotic membrane (AM). Methods. RT-PCR was used to examine the expression of ...
596 citations
TL;DR: Amniotic membrane matrix uniquely suppresses TGF-ß signaling in both types of fibroblasts and may also suppress signaling via CD44, ß1 integrin and FGFR1/ flg, which means the phenotype may become less mitogenic, contractile and fibrogenic.
Abstract: Purpose. When used as an alternative substrate following bare sclera removal of pterygium and other ocular surface diseases, amniotic membrane transplantation can reduce scarring on the reconstruct...
289 citations
TL;DR: Compared with an oxidative stress model of cells treated with 0.001–0.000001% hydrogen peroxide, timolol-BAC(+) induced a rapid decrease in cell viability ranging from 40% immediately after treatment to 85% 24h later and a small initial decrease in Cell viability was also observed with all tested concentrations of timolo...
Abstract: Purpose. To compare the toxicity of a short-time application of timolol with benzalkonium chloride (timolol-BAC+) and unpreserved timolol (timolol-BAC-) in a human conjunctival cell line. Methods. ...
159 citations
TL;DR: The new equation for the pressure-volume relation derived from all the currently available ocular rigidity data on the living human eye gives a larger volume increment for a given increment of pressure than Friedenwald's equation based on measurements performed on cadaver eyes.
Abstract: Purpose. The pressure-volume relation for an eye is the mathematical equation that relates changes in intraocular pressure to changes in intraocular volume. This relation is useful for calculating outflow facility from tonography and pulsatile ocular blood flow from intraocular pressure pulsations. The present work develops a new relation by culling together all the published direct manometric rigidity measurements on living human eyes. Methods. A total of 182 data items taken from 21 eyes are available in the 1958–62 literature of Ytteborg, Prijot, Eisenlohr, Langham and Maumenee. The approach was (i) to perform an error analysis based on the various experimental conditions, (ii) to assume general mathematical forms for the relation, (iii) to use least-squares analysis and statistical measures to find the optimal data representation, and (iv) to introduce the total volume of the eye into the formulation. Results. A new formula for the pressure-volume relation for the living human eye is derived relating ...
158 citations
TL;DR: On-line analysis of the pupillogram using the suggested parameters can be performed easily to produce a real-time assessment of an individual's state of alertness or fatigue that correlates with his/her subjective assessment of this state.
Abstract: PURPOSE: To determine whether parameters calculated from pupillary activity can identify subjects with sleep deprivation, and whether the objective values correlate with a subjective feeling of fatigue. METHODS: pupil size in the dark was recorded continuously for 10 minutes in 12 healthy volunteers using an infrared video camera. Two recordings were made for each subject: after a full night's sleep, and after 24 hours of sleep deprivation. Several parameters calculated from pupil size and activity were analyzed and compared with a subjective rating of the state of alertness provided by the participants in each test. RESULTS: All pupillary parameters differed significantly between alertness and fatigue (p = 0.0076-0. 0186). Changes in one of the parameters - average pupillary diameter - correlated with changes in the subjective level of sleepiness (r = -0.51, p = 0.028). Although the values of most parameters differed among subjects, an absolute value of more than 25 in one parameter, cumulative pupillary variability ratio, was always associated with sleep deprivation. CONCLUSION: On-line analysis of the pupillogram using the suggested parameters can be performed easily to produce a real-time assessment of an individual's state of alertness or fatigue that correlates with his/her subjective assessment of this state. Language: en
128 citations
TL;DR: This study demonstrates that, in healthy subjects, pCO(2) is an important determinant of foveal choroidal blood flow, whereas pO (2) has little impact on it.
Abstract: Purpose. Arterial carbon dioxide tension and arterial oxygen tension are important determinants of retinal and cerebral blood flow. In the present study the hypothesis that changes in arterial blood gases also influence choroidal blood flow was tested. Methods. The effect of breathing different mixtures of oxygen (O 2) and carbon dioxide (CO 2) on choroidal blood flow in the foveal region was investigated in healthy subjects. The study was performed in a randomized, double-masked four way cross-over design in 16 subjects. Using a compact laser Doppler flowmeter, red blood cell velocity (ChBVel), volume (ChBVol), and flow (ChBF) in the choroidal vasculature were measured during the breathing of various mixtures of O 2 and CO 2 (hyperoxia-hypercapnia): 100% O 2, 97% O 2 + 3% CO 2, 95% O 2 + 5% CO 2 (carbogen) and 92% O 2 + 8% CO 2. Arterial oxygen tension (pO 2) and carbon dioxide tension (pCO 2) were measured from arterialized blood samples from the earlobe. Results. Breathing 100% O 2 had no significant e...
119 citations
TL;DR: The ability of aminoguanidine to normalize the hyperglycemia-induced increases in retinal oxidative stress, NO and PKC in diabetic rats and galactose-fed rats suggests that these abnormalities may be inter-related in the retina, and that the biochemical mechanism by which aminogsuanidine inhibits retinal microvascular disease in diabetes may be complex.
Abstract: Purpose. Aminoguanidine has been found to inhibit the development of some retinal lesions in diabetic rats and diabetic dogs, thereby raising a possibility that the formation of glycation end products (AGEs) may be an essential step in the pathogenesis of the retinopathy. The purpose of this study is to investigate the effect of aminoguanidine administration on other metabolic abnormalities which might be involved in the development of retinopathy in two models of the retinopathy, alloxan diabetes and experimental galactosemia. Methods. Oxidative stress, nitric oxide (NO) and the activity of protein kinase C (PKC, total activity) were measured in the retina of the rats experimentally diabetic or galactosemic for 2 months. Effect of aminoguanidine administration on the inhibition of hyperglycemia-induced retinal dysmetabolism was investigated. Results. Two months of diabetes or experimental galactosemia in rats resulted in elevation of retinal oxidative stress (increase in thiobarbituric acid reactive subs...
114 citations
TL;DR: Increased vascular permeability for plasma proteins induced by VEGF in blood-retinal barrier endothelium is predominantly caused by a mechanism involving active trans-endothelial transport via pinocytotic vesicles and not by formation of endothelial fenestrations or VVOs.
Abstract: Purpose. In tissues outside the brain, vascular endothelial growth factor-A (VEGF) causes vascular hyper-permeability by opening of inter-endothelial junctions and induction of fenestrations and vesiculo-vacuolar organelles (VVOs). In preliminary studies, we observed that in blood-retinal barrier endothelium, other cellular mechanisms may underlie increased permeability caused by VEGF. This was further investigated in material of an in vivo experimental model of VEGF-induced retinopathy. Methods. Two monkeys received 4 intravitreal injections of 0.5 µg VEGF in one eye and PBS in the other eye prior to sacrifice at day 9. One monkey received 12 injections of 1.25 µg VEGF in one eye and PBS in the other eye prior to sacrifice at day 24. As a control, an untreated eye of a fourth monkey was studied. Results. In the high-dose VEGF-injected eye, fluorescein angiography showed intense retinal micro-vascular leakage. This leakage was also demonstrated by immunohistochemistry demonstrating extravasation of endoge...
110 citations
TL;DR: It is suggested that diabetes affects the retrograde axonal transport in large- and medium-sized retinal ganglion cells despite the absence of morphological changes in the perikaryon and decrease in total cell population.
Abstract: Purpose. Several abnormalities in visual pathway functions in diabetic humans and animals have been reported. We demonstrated retrograde axonal transport impairment in retinal ganglion cells of streptozotocin-diabetic rats. Methods. Diabetes was induced in male Wistar albino rats by intraperitoneal injection of streptozotocin. Three months after the induction of diabetes, fluoro-gold was injected into the dorsal lateral geniculate nucleus. Percentages of fluoro-gold-labeled large-, medium- and small-sized retinal ganglion cells per total population were calculated in wholemount retinas of diabetic and control rats. The same sections were stained with cresyl violet and each retinal ganglion cell type evaluated by light microscopy. Results. Although a quantitative decrease in the population of each retinal ganglion cell type was not observed, mean percentages of fluoro-gold-labeled large- and medium-sized retinal ganglion cells per total population were significantly decreased in diabetic rats compared with...
77 citations
TL;DR: The results indicate that cataract formation in diabetes involves both glycation and oxidative stress processes and the positive effect of probucol on cysteine/GSH metabolism imbalance indicates that derangements of one of the retinal defense systems against oxidative stress can be normalized by antioxidants.
Abstract: Purpose. To study effects of inhibition of glycation, and oxidative stress on the development of cataract and retinal vessel abnormalities in diabetic rats. Methods. Diabetes was induced in male Wistar rats with streptozocin (STZ; 60 mg/kg BW, i.p.). Diabetic as well as strain matched control rats were fed 1) a normal diet, 2) addition of aminoguanidine in the drinking water (0.5 g/l for diabetic rats and 1.0 g/l for control rats) or 3) probucol in the pellets (1% w/w). After 6 months, the number of acellular vessels, endothelial cells and pericytes were counted in trypsin digested retinal vessel preparations, and the total retinal tissue amount of glutathione (GSH) and cysteine was measured with HPLC. Results. Cataract formation occurred after 13 weeks in diabetic animals compared with 17 weeks for those treated with aminoguanidine, and 16 weeks for those treated with probucol (p < 0.001 in both cases). Aminoguanidine inhibited the formation of acellular collapsed capillary strands, 9 (3–14) vs. 18 (1...
76 citations
TL;DR: The results suggest that calpain isoforms may play an important role in neuronal cell death induced by retinal ischemia-reperfusion injury in rat.
Abstract: Purpose. Much evidence has accumulated suggesting that activation of calpain causes neuronal cell death in ischemic brain. However, little is known about the involvement of calpain in retinal cell death in ischemic injury. Thus, the purpose of present study was to investigate the involvement of calpain isoforms (m- and µ-calpain) in ischemia-reperfusion injury in retina from rat. Methods. Retinal ischemia was produced by occlusion of the central retinal artery for one hour, and this was followed by reperfusion for seven days. Calpain mRNAs, calpain activities, total calcium content and proteolysis of a-spectrin were determined in retina. Effect of a calpain inhibitor SJA6017 was histologically tested in retinal injury after ischemia-reperfusion. Results. Following retinal ischemia, most of cells in the ganglion cell layer were sloughed off by day 1 after reperfusion, followed by loss of cells in the inner plexiform layer on day 3 and loss of cells in the inner nuclear layer by day 5. These morphologic cha...
TL;DR: Support is shown for a role for MMP-2 in the development of CNV in age-related macular degeneration through increased mRNA expression in experimental choroidal neovascularization.
Abstract: Purpose. Matrix metalloproteinases (MMP) are a family of proteolytic enzymes that degrade basement membrane and extracellular matrix proteins. To gain information on the possible role of MMPs in ch...
TL;DR: The data demonstrate an influence of macrophage depletion on the course of HSV-1 keratitis in mice and influence the viral replication in the cornea and the immune-mediated process of HSK.
Abstract: Purpose. Herpes simplex virus type 1 infection of the cornea induces an immune-mediated disease termed “herpes stromal keratitis” (HSK) that is a major cause of blindness. In this study we investigated the influence of macrophage depletion by Cl 2 MDP encapsulated in liposomes (Cl 2 MDP-LIP) on the course of HSV-1 keratitis. Methods. The corneas of BALB/c mice were infected with 10 5 PFU of HSV-1 (KOS strain). Mice groups received sub-conjunctival PBS or Cl 2 MDP-LIP injections 7 and 2 days prior to infection. The eyes were studied clinically, histologically and immunohistochemically with F4/80 antibody at various time points after treatment. Clearance of the virus from the HSV-infected eyes was measured with a standard plaque assay. Results. After subconjunctival Cl 2 MDP-LIP treatment, the HSV-1-induced epithelial keratitis was more severe (P < 0.05). The virus titers were significantly higher after macrophage depletion (day 7, P < 0.005). Stromal keratitis developed in 78.6% of HSV-1 infected PBS...
TL;DR: In comparison to healthy controls, patients with vasospastic NTG seem to over- as well as under-express certain genes in their lymphocytes, particularly those coding for p53-protein, NTP and 20 S proteasome subunit XAPC7.
Abstract: Background. In glaucoma there is a loss of retinal ganglion cells. There is evidence that this loss can occur by apoptosis. The signal transduction leading to retinal ganglion cell apoptosis in gla...
TL;DR: The observation that MT1-MMP expression may be up-regulated in keratoconus corneas, taken together with the demonstration that human corneal cells can express this enzyme, which in turn can activate latent MMP-2, provide evidence for a possible role for MT1/MMP in the pathogenesis of keratconus.
Abstract: Purpose. To determine whether MT1-MMP and MMP-2 are expressed in normal and keratoconic corneas, and to investigate the ability of MT1-MMP, expressed on cultured keratocytes after stimulation with concanavalin A, to activate pro-gelatinase A (pro-MMP 2). Methods. Specimens of keratoconus corneas (n = 20), removed at corneal transplantation, were obtained from pathology archives, sections were cut, and were stained with an antibody to MT1-MMP, using peroxidase immunohistochemistry. Eye banked corneas served as controls (n = 14). Normal human keratocyte cultures were initiated from eye bank corneas, and after stimulation with con A, MMPs in the media were examined using gelatin zymography and immunoblotting, and MT1-MMP expression was analysed by flow cytometry and immunoblotting. Results. All corneas showed some expression of MT1-MMP and MMP-2, although the degree of staining varied greatly. The MMPs were present in the epithelium, endothelium and stroma. Expression of MT1-MMP, but not MMP-2, in the epithelium and stroma, was significantly elevated in keratoconus, compared to normal corneas. In vitro, keratocytes stimulated with con A expressed MT1-MMP and produced active MMP-2, detected by zymography. These responses to con A were concentration-dependent and MT1-MMP expression and MMP-2 activation correlated significantly (p = 0.0003) In addition, MMP inhibitors abolished MMP-2 activation, providing further evidence that MT1-MMP activated MMP-2. Conclusion. The observation that MT1-MMP expression may be up-regulated in keratoconus corneas, taken together with the demonstration that human corneal cells can express this enzyme, which in turn can activate latent MMP-2, provide evidence for a possible role for MT1-MMP in the pathogenesis of keratoconus.
TL;DR: The clearest image with the retraced method for endothelial cell analysis with the IMAGEnet system is suggested, as the variability in hexagonality results between the first and the clearest images can be clinically significant.
Abstract: Purpose. Corneal endothelium plays an important role in the maintenance of corneal transparency and stability. The purpose of this study was to evaluate the performance of the TOPCON SP-2000P and IMAGEnet system in terms of (a) the difference in results between automated endothelial cell analysis and retraced cell analysis, (b) the differences in the endothelial cell analysis when using the first image and the clearest of three images taken with the SP-2000P, (c) the repeatability and reproducibility of the IMAGEnet system in the determination of corneal endothelial cell variables, and (d) the repeatability and reproducibility of the SP-2000P in capturing endothelial cell images for endothelial cell analysis. Methods. Two experiments were performed. Twenty male subjects participated in the first experiment in which endothelial images were captured by examiner SWC and endothelial cell analysis was performed by two examiners, SWC (twice on two different days) and PC (once only). Nineteen male subjects parti...
TL;DR: The predominant expression of CTGF mRNA in human ASC and human PCO membranes suggests a significant role ofCTGF in the pathological course of these ocular disorders.
Abstract: Purpose. To investigate the correlation between connective tissue growth factor (CTGF) mRNA expression and immunohistochemical characteristics of anterior subcapsular cataract (ASC) formation as well as posterior capsule opacification (PCO) development (expression of type I collagen, a-smooth muscle actin and tenascin) under in vivo and under in vitro conditions in human and porcine lens epithelial cells. Methods. CTGF mRNA expression was investigated using in situ hybridization and RT-PCR. Expression of type I collagen, a-smooth muscle actin and tenascin was detected by immunohistochemical staining. Results. CTGF mRNA was expressed in human cataractous plaques of ASC and human PCO membranes, and appeared simultanously with the expression of type I collagen, a-smooth muscle actin and tenascin. Conclusion. The predominant expression of CTGF mRNA in human ASC and human PCO membranes suggests a significant role of CTGF in the pathological course of these ocular disorders.
TL;DR: Melatonin may protect against the UVB-induced cataract development by directly quenching lipid peroxides and indirectly by enhancing the production of the endogenous antioxidant GSH.
Abstract: Purpose. To investigate the defensive effect of melatonin against oxidative stresses in ultraviolet-B (UVB) radiation induced cataract development. Methods. Young rats received 8 kJ/m 2 UVB for 15 min. For the intervention of cataract development intraperitoneal injection of melatonin (4 mg/kg daily for 1 week) following UVB exposure was performed. Lenticular glutathione peroxidase (GSHPx), catalase (CAT), superoxide dismutase (SOD) activities and glutathione (GSH), malondialdehyde (MDA) levels were determined in UVB-melatonin, UVB, and control groups. Results. One week after exposure in the UVB group, lens opacities were observed and CAT, SOD, and GSHPx activities, and GSH level were lower than control and MDA level was higher than control (p < 0.05). In the UVB-melatonin group CAT and SOD activities were lower than control (p < 0.05), and the MDA level was lower than the UVB group (p < 0.05). Conclusions. These results suggest that melatonin may protect against the UVB-induced cataract developm...
TL;DR: This study provides the first report on the expression and function of an organic cation transporter, OCT3, in the eye and in particular the RPE and it is likely that OCT3 participates in the clearance of dopamine and histamine from the subretinal space and may also play a key role in the disposition of the retinal neurotoxin MPP +.
Abstract: Purpose. Dopamine has several important functions in the retina including a possible role in controlling photoreceptor disk shedding to the RPE. While some cells express a transporter for dopamine, the RPE cell does not, leading us to ask whether the newly described catecholamine transport system, the extraneuronal monoamine transporter (uptake 2) (also known as organic cation transporter 3 (OCT3), is present in RPE and might function as a transporter for dopamine. OCT3 also accepts histamine as a transportable substrate and so we investigated the interaction of this retinal neurotransmitter with OCT3. Methods. OCT3 expression in the mouse eye was analyzed by in situ hybridization, Northern blot analysis and RT-PCR. OCT3 function was analyzed in cultured human ARPE-19 cells by monitoring the uptake of 1-methyl-4-phenyl pyridinium (MPP +) , a neurotoxin, which is a known substrate for OCT3. Results. In situ hybridization analysis showed that OCT3 is expressed in mouse RPE and in several cell types of the n...
TL;DR: The data suggests that the high levels of MMPs found in patients with chronic uveitis might contribute to the damage often seen in these eyes.
Abstract: Purpose. Matrix metalloproteinases have been shown to play a role in active uveitis. Transcription of MMPs is induced by a number of growth factors and cytokines. This study investigates the role of MMPs in chronic uveitis and correlates the amounts of MMP-2 and -9 in intraocular samples to the intraocular levels of proinflammatory cytokines (Interleukin-1 [IL-1], Interleukin-12 [IL-12] and Interleukin-1 receptor antagonist [IL-1ra]). Methods. Aqueous humor of 16 patients was collected during surgical or diagnostic procedures (control group: cataract patients). MMP-2 and -9 were measured using zymography. IL-1s, IL-12 and IL-1ra were evaluated by ELISA. Results. We found MMP-2 and -9 in all of our uveitis patients. In the control group only MMP-2 was seen. Higher levels of MMP-2 and -9 were found in patients with higher activity of uveitis (p < 0.014 for MMP-2, p < 0.0054 for MMP-9). The amounts of IL-1s, IL-12 and IL-1ra detected in our uveitis patients correlated with levels of MMP-2 (p < 0.07,...
TL;DR: This model of retinal changes with age and diet in high-fat-fed C57BL/6 mice, a mouse model of human atherosclerosis, is suggested to prove useful for investigations into mechanisms on the effects of diet on the RPE and Bruch’s membrane.
Abstract: Purpose. Due to a possible link in humans between atherosclerosis, a high-fat diet, and the development of age-related retinal degenerations, we investigated retinal changes with age and diet in high-fat-fed C57BL/6 mice, a mouse model of human atherosclerosis. Methods. We fed C57BL/6J mice either a normal chow diet or an atherogenic diet containing 15% total fat for either 15, 30 or 45 weeks. We sacrificed the animals and examined the eyes using fluorescence microscopy, light microscopy and transmission electron microscopy. Results. At 15 weeks, in high-fat-fed mice, there was an increase in the number of autofluorescent granules in the retinal pigment epithelium (RPE) compared to chow-fed mice. By light microscopy, we noted no remarkable differences in the inner retinal layers between mice fed either diet for 15 or 30 weeks. In contrast, we noted major changes in transmission electron micrographs from the 30 week high-fat group. In the RPE, these included: (1) an increase in the number and size of autop...
TL;DR: Differences in CT measurements obtained from the first and clearest images captured by the SP-2000P were not statistically significant but can be clinically significant for peripheral CT, which has relevance to the measurement of the cornea before and after refractive surgery.
Abstract: Purpose. Modern refractive surgery and follow up relies on a knowledge of corneal thickness (CT) and shape, and the reliability of modern instrumentation providing such data is important. This study sought to determine the performance of the TOPCON SP-2000P specular microscope in measuring CT. The aims of this study were: (a) to determine if there is any difference between the CT results obtained from the first image, the clearest of three images and the mean of measurements from three images; (b) to determine the correlation between central and peripheral CT and (c) to investigate the reliability (repeatability and reproducibility) of the SP-2000P in the determination of central and peripheral CT. Methods. The central and peripheral CT measurements shown on the first (f), the clearest (c) of three images and the mean (m) CT shown in three images captured with the SP-2000P of 43 subjects were compared. All images were captured by the same examiner. Nineteen of the subjects (male) returned on another day a...
TL;DR: BBR measured under different conditions produced different results so it is important for investigators to describe the baseline condition very clearly in studies where BBR are measured, and results indicated that mean blink rate was affected by the position of gaze and not the level of task difficulty.
Abstract: Purpose. To compare the baseline blink rates (BBR) measured under different (baseline) conditions, and to compare the blink rates measured when performing two visual tasks of different levels of difficulty at two positions of gaze. Methods. In the first single masked experiment, BBR were measured under three different conditions – (i) conversation with the subjects (ii) taking the visual acuity and (iii) keeping the subjects waiting in an empty examination room. In the second single masked experiment, the subjects were required to perform an easy task (reading normal English words) and a difficult task (reading mirror-image English words) at primary gaze and down gaze. The orders of the tasks performed were randomly allocated. Results. BBR taken under the three different conditions were significantly different from each other. No significant difference was found between the blink rates measured when performing the two reading tasks if they were performed at the same position of gaze. The mean blink rates ...
TL;DR: Results indicate that conjunctival inflammation influences the corneal immunological environment, and may affect the fate of orthotopicCorneal allografts.
Abstract: Purpose. The virtual absence of Langerhans cells (LC) in donor or recipient corneal epithelium is known to be an important factor in the acceptance of orthotopic corneal allografts. Though it is well known that various types of stimulation to the cornea induce LC migration into the corneal epithelium, resulting in poor graft survival, the influence of conjunctival inflammation on LC migration into the cornea has not been elucidated. Therefore we examined whether LCs migrate into the cornea in the presence of conjunctival inflammation. Methods. Sixteen BALB/c mice were divided into four groups. Group A: 4 mice with corneal inflammation induced by two 9–0 silk interrupted sutures in the central cornea (positive control); Group B: 4 mice with conjunctival inflammation induced by two 9–0 silk interrupted sutures in the temporal and nasal bulbar conjunctiva 1 mm from the limbus; Group C: 4 mice with conjunctival inflammation by two 10–0 nylon interrupted sutures in the temporal and nasal bulbar conjunctiva 1 m...
TL;DR: Data indicate that activation of P2Y 2 receptor increases tear fluid secretion accompanied with some proteins in normal rabbits, indicating that the purinergic agonists, UTP and ATP, have potential therapeutic value in the treatment of dry eye.
Abstract: Purpose. The purinergic P2Y 2 receptor agonists stimulate active Cl - transport across the excised rabbit conjunctival tissue in vitro. We determined whether UTP or ATP could increase the tear volume and change tear fluid composition in normal rabbits in vivo. Methods. Fifty µL was applied to rabbit eyes of UTP, ATP at concentrations of 0.1, 0.3, 1, 3, 8.5% (1.8–154 mM) or saline. A modified Schirmer test with topical anesthesia was performed 5, 15, 30 and 60 min after the instillation. In studies lasting 30 days, 50 µL of 0.5% UTP was applied 6 times a day for 4 weeks. Tear samples were collected from the conjunctival sac with a glass microcapillary. The protein profile of the tear fluid was analyzed by SDS-polyacrylamide gel electrophoresis and total protein was measured with the Bradford assay. The Easy-Titer rabbit IgG assay kit was used for the determination of immunoglobulin G (IgG). Results. UTP had dose-dependent stimulatory effects on tear secretion. It maximally increased tear secretion about 4-...
TL;DR: In this article, the authors used specific immunoblot assay (IBA) to determine any correlation between activity of ocular cicatricial pemphigoid and titer of anti-s4 antibodies, and any effect of intravenous immunoglobulin (IVIg) therapy on serum levels of anti s4 antibodies followed over a 12 month period.
Abstract: Purpose. To determine any correlation between activity of ocular cicatricial pemphigoid and titer of anti-s4 antibodies, and any effect of intravenous immunoglobulin (IVIg) therapy on serum levels of anti-s4 antibodies followed over a 12 month period, using the specific immunoblot assay (IBA). Patients and methods. Eight patients diagnosed with OCP and treated with IVIg as monotherapy were included in the study. Each patient was treated with at least two immunosuppressive agents prior to the institution of IVIg. The presence of anti-s4 antibodies in the patients’ sera was detected by IBA using bovine gingival lysate (BGL) or tumor cell line lysate (TCL) as substrates. The activity of OCP was graded based on the conjunctival injection using a scale of zero to four in increments of 0.5 at monthly intervals. To determine the correlation between serum levels of circulating autoantibody and the patients’ conjunctival disease activity, the titer of anti-s4 antibodies was determined at monthly intervals during t...
TL;DR: The therapeutic effect of panretinal photocoagulation on PDR might be partly exerted by reduction of the levels of VEGF and HGF in ocular fluid, since these growth factors might influence the progression of retinopathy in different ways.
Abstract: Purpose To investigate clinical factors related to the aqueous humor levels of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) in patients with proliferative diabetic
TL;DR: An N-terminal sequence of KE may be related to formation of amyloid associated with R124 mutations in three corneal dystrophies harboring mutations at Arg-124 in the BIGH3 gene.
Abstract: Purpose. To investigate corneal deposits associated with kerato-epithelin (KE) in three corneal dystrophies harboring mutations at Arg-124 in the BIGH3 gene. Methods. Six patients with Avellino corneal dystrophy (ACD) associated with R124H, one patient with superficial granular corneal dystrophy (SGCD) associated with R124L, and seven patients with lattice corneal dystrophy type 1 (CDL1) associated with R124C were examined. Corneal buttons obtained during keratoplasties were stained with Masson's trichrome and with Congo red, and immunostained with antibodies specific for N-terminal and C-terminal portions of KE (KE-15 and KE-2, respectively). Results. In all corneas with ACD, subepithelial to midstromal deposits of granular material stained with KE-2 and KE-15. However, deep stromal deposits containing amyloid reacted with KE-2, but not KE-15. Granular deposits in the subepithelial layer observed in SGCD stained intensely with KE-2 and KE-15. In all corneas with CDL1, subepithelial and midstromal amyloid...
TL;DR: The results indicate that the isolated cells were vascular endothelial cells showing both morphologic and functional characteristics of retinal vascular endothelium, indicating that the magnetic-bead technique was useful for isolating high purity RECs that can be cultured to study the physiological, immunological and biochemical role of the endothelia in various ocular diseases.
Abstract: Purpose. To isolate retinal endothelial cells (RECs) from Lewis rats using magnetic beads coated with antibodies to rat platelet-endothelial cell adhesion molecule-1 (PECAM-1) and to characterize the cultured RECs. Methods. Magnetic beads were coated with anti-rat PECAM-1 antibodies. Retinas were obtained from Lewis rat eyes. After the retinas were digested with collagenase, they were incubated with the antibody-coated beads with agitation. RECs that stuck to the beads were collected with a magnetic particle concentrator and cultured in fibronectin coated wells. The characteristics of the RECs were examined by immunohistochemical study utilizing von Willebrand’s Factor, acetylated low-density lipoprotein uptake and transmission electron microscopy. Results. The cells isolated using the PECAM-1-coated magnetic-bead technique formed a contact-inhibited cobblestone monolayer that stained positive for von Willebrand’s Factor. These cells revealed low-density lipoprotein uptake. Ultrastructurally, the isolated...
TL;DR: A clear advantage to digesting the extracellular matrix of glaucomatous TM tissue to obtain sufficient numbers of healthy cells for use in experiments is demonstrated.
Abstract: Purpose. Isolation and culture of human trabecular meshwork (TM) cells from primary open-angle glaucomatous (POAG) tissue has proven difficult. The objective of this study was to directly compare t...