Otto Schmut

The organization of tissues of the eye by different collagen types

Bovine cornea, sclera, iris, ciliary body, choroid, zonular fibers, lens capsule, lens nucleus, vitreous body, and retina were investigated for collagen content and type. Cornea, sclera, iris, ciliary body, choroid, lens capsule, and vitreous body contain hydroxyproline, whereas in zonular fibers, lens nucleus, and retina no hydroxyproline was detectable. Preparative isolation of collagen was achieved by digestion of the different eye tissues with pepsin. The pepsin-solubilized collagen was separated by differential salt precipitation into different collagen types. The polyacrylamide gel electrophoresis of the pepsin-solubilized collagens revealed type I collagen in cornea, sclera, iris, ciliary body, and choroid. As well as type I collagen, type III collagen was isolated from cornea, sclera, and uveal tissues. The identification of types I and III collagen was supported by the CNBr-derived peptides of these collagens. Lens capsule collagen consisted mainly of type IV collagen. Zonular fibers contained no hydroxyproline but when examined by polyacrylamide gel electrophoresis, a band migrating in the α-position of collagen was observed. Polyacrylamide gel electrophoresis of both the pepsinsolubilized component and the CNBr-derived peptides of vitreous body protein showed no relation to any of the four common collagen types. Hornhaut, Lederhaut, Regenbogenhaut, Ziliarkörper, Aderhaut, Zonulafasern, Linsenkapsel, Linsenkern, Glaskörper und Netzhaut werden auf ihren Kollagengehalt untersucht. Auch soll festgestellt werden, welche Kollagentypen diese Gewebe aufbauen. Hornhaut, Lederhaut, Regenbogenhaut, Ziliarkörper, Aderhaut, Linsenkapsel und Glaskörper enthalten Hydroxyprolin. Hingegen ist in Zonulafasern, Linsenkern und Netzhaut kein Hydroxyprolin nachzuweisen. Aus den verschiedenen Augengeweben wurde mit Pepsin Kollagen gelöst, das durch Präzipitation mit NaCl-Lösungen verschiedener Molarität in verschiedene Kollagentypen getrennt werden kann. Die Polyacrylamidgel-Elektrophorese des pepsin-gelösten Kollagens zeigte Type I Kollagen in Hornhaut, Lederhaut, Regenbogenhaut, Ziliarkörper und Aderhaut. Neben Typ I Kollagen wurde aus Hornhaut, Lederhaut und den Geweben der Uvea Typ III Kollagen isoliert. Typ I und Typ III Kollagen wurden durch die Peptide, die man mit CNBr-Spaltung erhalten kann, identifiziert. Die Linsenkapsel besteht hauptsächlich aus Typ IV Kollagen. Zonulafasern enthalten kein Hydroxyprolin, zeigen aber in der Polyacrylamidgel-Elektrophorese eine Farblinie, die im α-Bereich von Kollagen wandert. Die Polyacrylamidgel-Elektrophorese des pepsin-gelösten Kollagens des Glaskörpers sowie die CNBr-Spaltprodukte dieses Kollagens zeigten keine Identität mit einem der vier bisher isolierten Kollagentypen.

The redox state of human serum albumin in eye diseases with and without complications

Purpose:u2002 To investigate the redox state of human serum albumin concerning cysteine‐34 as a possible systemic redox marker in patients with different eye diseases with and without complications and with consideration of possible effects of age.

Role of Inflammation-Related Gene Polymorphisms in Patients with Central Retinal Vein Occlusion

OBJECTIVEnCentral retinal vein occlusion (CRVO) is a vision-threatening disease, primarily occurring among patients aged more than 60 years. Several risk factors, including arterial hypertension and diabetes mellitus, have been identified. Compression of the central retinal vein by an atherosclerotic retinal artery at the lamina cribrosa also has been implicated in the pathogenesis of the disease. Functional gene polymorphisms of cytokines or chemokines previously shown to affect atherogenesis or hemostasis are potential risk factors for CRVO. The present study investigates a hypothesized association between inflammation-related gene polymorphisms and the presence of CRVO in a relatively large cohort of patients.nnnDESIGNnCase-control study.nnnPARTICIPANTSnThe study group consisted of 315 patients with CRVO and 335 control subjects.nnnMETHODSnDetermination of genotypes was done by 5 exonuclease assay (TaqMan).nnnMAIN OUTCOME MEASURESnGenotypes of interleukin (IL)1β -511C>T, IL1 receptor antagonist (IL1RN) 1018T>C, IL4 -584C>T, IL6 -174G>C, IL10 -592C>A, IL18 183A>G, tumor necrosis factor (TNF)-α -308G>A, monocyte chemoattractant protein (MCP)-1/CCL2 -2518A>G, IL8 -251A>T, and RANTES (CCL5) -403G>A polymorphisms.nnnRESULTSnGenotype distributions and allele frequencies of the investigated gene polymorphisms did not significantly differ between both groups (P>0.05). Arterial hypertension, diabetes mellitus, and cigarette smoking were significantly more frequent in patients with CRVO than among control subjects (arterial hypertension: 67.0% vs. 52.2%, P<0.001; diabetes mellitus: 16.8% vs. 6.3%, P<0.001, cigarette smoking: 32.1% vs. 23.6%, P = 0.02). In a logistic regression analysis, the presence of arterial hypertension was associated with an odds ratio (OR) of 1.75 (95% confidence interval [CI], 1.26-2.44) in those with CRVO, whereas an OR of 2.52 (95% CI, 1.46-4.35) was found in those with diabetes mellitus. A history of cigarette smoking was associated with an OR of 1.57 (95% CI, 1.09 - 2.25) for CRVO.nnnCONCLUSIONSnOur data suggest that the investigated inflammation-related gene polymorphisms are unlikely major risk factors for CRVO.nnnFINANCIAL DISCLOSURE(S)nThe author(s) have no proprietary or commercial interest in any materials discussed in this article.

[Glaucoma and oxidative stress. Determination of malondialdehyde--a product of lipid peroxidation].

ZusammenfassungHintergrund Das primäre Offenwinkelglaukom (POWG) ist eine Erkrankung multifaktorieller Genese, wobei Altern, Rassenzugehörigkeit, genetische Disposition, vaskulärer Status, möglicherweise Entzündung, aber auch oxidativer Stress eine Rolle spielen können. Wir gingen der Frage nach, ob sich mit Hilfe eines gängigen Tests betreffend eines oxidativen Stressprodukts Unterschiede im Serum und im Kammerwasser von Patienten mit POWG im Vergleich zu jenen ohne POWG finden lassen.Patienten und Methode Untersucht wurde Kammerwasser von 33xa0Patienten mit POWG, 111xa0Patienten ohne POWG mit Katarakt, 39 ohne POWG mit Katarakt und Pseudoexfoliationssyndrom sowie Serum von allen 3 genannten Gruppen und zusätzlich von 43 augengesunden Probanden. Als Methode wurde der Nachweis des Stressprodukts Malondialdehyd, entstehend aus Peroxidation von Lipiden, mit Hilfe der Thiobarbitursäure angewendet.Ergebnisse Thiobarbitursäurereaktive Substanzen (TBARS) werden in µmol/l angegeben: 1) im Serum der Gruppe Katarakt 1.176, der Gruppe Katarakt und Pseudoexfoliation 1.019, der Gruppe mit primärem Offenwinkelglaukom 0.992 und der augengesunden Gruppe 0.983. 2) im Kammerwasser der Gruppe Katarakt 0.279, der Gruppe Katarakt und Pseudoexfoliation 0.274 sowie der Gruppe mit primärem Offenwinkelglaukom 0.298. Es fanden sich weder im Kammerwasser noch im Serum von Patienten mit POWG im Vergleich zu jenen ohne POWG statistisch signifikante Unterschiede im Gehalt von TBARS. Es zeigte sich jedoch eine signifikante positive Korrelation zwischen den Werten im Serum und jenen im Kammerwasser.Schlussfolgerung Erstmalig wurde mit der TBARS-Methode Malondialdehyd im Kammerwasser von Glaukomaugen nachgewiesen. Es ließen sich keine Unterschiede betreffend dieses oxidativen Stressprodukts in Kammerwasser und Serum von Patienten mit POWG bzw. ohne POWG nachweisen. Zu bedenken wäre, dass jene Gruppe ohne POWG an Katarakt erkrankt war, welche per se eine Erkrankung mitbedingt durch oxidativen Stress ist.AbstractBackground Primary open-angle glaucoma (POAG) is a multifactorial disease, in which aging, race, genetic disposition, vascular status, probably inflammation, and maybe oxidative stress may play a specific role. We wanted to know if by using a common test for a specific oxidative stress product differences would be found in the aqueous humor and serum of patients with POAG, cataract without pseudoexfoliation, cataract with pseudoexfoliation, and volunteers without ocular disease.Patients and methods We examined the aqueous humor of 33 patients with POAG, 111 patients with cataract without pseudoexfoliation, 39 patients with cataract and pseudoexfoliation syndrome and the serum of all three groups plus of 43 volunteers without ocular disease. Malondialdehyde as an oxidative stress product of peroxidation of lipids was proven by thiobarbituric acid-reacting substances (TBARS).Results The amount of TBARS is given in µmol/l: (1) in the serum of the group with cataract 1.176, with cataract and pseudoexfoliation 1.019, with POAG 0.992, and with healthy eyes 0.983; (2) in the aqueous humor of the group with cataract 0.279, with cataract and pseudoexfoliation 0.274, and with POAG 0.298. There were no statistically significant differences of TBARS (p<0.05) in either the aqueous humor or in the serum of patients with POAG in comparison to those patients without POAG. However, there was a significantly positive correlation between the values in the serum and the aqueous humor.Conclusion For the first time malondialdehyde as a product of lipid peroxidation was determined in the aqueous humor of glaucomatous eyes. With the TBARS method used in our study, it was not possible to detect statistically significant differences of the lipid peroxidation product malondialdehyde between patients with or without POAG. It has to be taken into account that the cataract of the patients in the control group might be a disease per se caused by at least a certain amount of oxidative stress and that these subjects might therefore not be ideal as a control group.

The effect of iodide iontophoresis on the antioxidative capacity of the tear fluid

BackgroundEnvironmental oxidative stress changing the properties of the tear fluid can lead to keratoconjunctivitis sicca (dry eye syndrome). The aim of this study was to determine whether iodide iontophoresis influences the antioxidative capacity (ACWxa0=xa0water soluble antioxidative capacity) of the tear fluid, and to compare iodide iontophoresis with other balneotherapeutic measures.MethodsThis prospective study evaluated 92 patients in four groups. Twenty-four patients were treated with iodide iontophoresis, 24 with other balneotherapeutic methods. Twenty-five patients received iodide iontophoresis combined with other balneotherapeutic methods and 21 persons received no treatment (control). Unstimulated tear fluid, serum and urine were collected. ACW was determined photochemically in tear fluid and serum; iodine was measured in urine photometrically.ResultsIodide iontophoresis increases the ACW of the tear fluid but not the ACW of the serum. Other iodine therapies increase the ACW in serum but not in tear fluid. Iodine excretion in urine was increased in all treated groups compared to the control.ConclusionThe increase of ACW in tear fluid after iodide iontophoresis can support the defense mechanism of the eye against oxidative influence effects, which may alleviate the symptoms of keratoconjunctivitis sicca.

Determination of uric acid concentrations in human tear fluid, aqueous humour and serum.

Purpose:u2002 The aim of this investigation was to assess and compare levels of uric acid in human tear fluid, aqueous humour and serum in cataract patients in order to obtain information about uric acid‐dependent antioxidative status.

Quantitative Bestimmung der Antiproteinasen in der menschlichen Tränenflüssigkeit

By quantitative determination of antiproteinases, i.e. α 1-antitrypsin, α 1-antichymotrypsin, and inter-α-trypsin inhibitor normal values could be obtained. The importance of antiproteinases is demonstrated in this paper. Especially α 1antitrypsin was very easy to quantitate and might be of diagnostic interest. Durch die quantitative Bestimmung der Antiproteinasen α 1Antitrypsin, α 1-Antichymotrypsin und Inter-α-Trypsin Inhibitor war es möglich, Normalwerte zu erstellen. In der vorliegenden Arbeit wird auf die Bedeutung der Antiproteinasen hingewiesen. Besonders α 1-Antitrypsin war am besten nachweisbar und dürfte von diagnostischem Interesse sein.

Iodid schützt Bindehautzellen vor der Schädigung durch UV-Licht

ZusammenfassungUltraviolette Strahlung ruft an Zellen des vorderen Augenabschnitts Schäden hervor, die mit Fotokeratitis, Carcinomata in situ der Bindehaut, Basaliomen an den Lidern, Melanomen des vorderen Augenabschnitts, dem Entstehen des Pterygiums und Pingueculae in Verbindung gebracht werden.Ein signifikanter, schützender Effekt durch Iodid gegen die Schädigung von Bindehautzellen durch UV-Licht wurde mit einem Zellanalysesystem (CASY-System) nachgewiesen.Eine Anreicherung von Iodid im äußeren Auge, die durch Balneotherapie und Ophthalmoiontophorese mit Iodsole in Bad Hall (Oberösterreich) erreicht wird, könnte einen Schutz des Auges gegen energiereiche Strahlung bewirken.SummaryUltraviolet (UV) radiation provokes damages of cells of the anterior parts of the eye which may be associated with photokeratitis, carcinomata in situ of the conjunctiva, basaliomas of the lids, melanomas of the anterior part of the eye, and the development of pterygia and pingueculae.A significant protective effect of iodide against the damage by UV-light of conjunctival cells was determined using a cell analysis system (CASY-system).Enrichment of iodide in the anterior parts of the eye, as obtained by balneotherapy or iontophoresis with iodine brine in Bad Hall (Upper Austria) may provide protection of the eye against high-energy radiation.

Pollen Enzymes Degrade Human Tear Fluid and Conjunctival Cells: An Approach to Understanding Seasonal Non-Allergic Conjunctivitis

BACKGROUNDnDuring pollen seasons, allergy-like symptoms can be observed in proven non-allergy sufferers. Pollen enzymes are thought to be responsible for conjunctival irritation. We investigated the influence of the well-known aggressive pollen species hazelnut (Corylus avellana) and birch pollen (Betula pendula) on both human tear fluid and conjunctival cell cultures. This study is an approach to seasonal non-allergic conjunctivitis (SNAC) syndrome.nnnMETHODSnZymography was carried out in order to investigate the proteolytic activity of the pollen. Thereafter, human tear fluid was incubated with pollen extract, and the results were studied by polyacrylamide gel electrophoresis. In addition, cultivated conjunctival cells (CHANG cells) were incubated with pollen extracts. Cytomorphological changes were analyzed using the CASY1 Cell Counter. Cell viability was quantified via MTS assay. The viability of the cells which were incubated with pollen extract was compared to the viability of control cells.nnnRESULTSnPollen proteases destroy tear fluid proteins, as observed by polyacrylamide gel electrophoresis. The treatment of CHANG cells with pollen extract induced a statistically significant decrease in cell viability, depending on the pollen extract concentration and the incubation period.nnnCONCLUSIONnEvidence of the destruction of tear fluid proteins and damage to human conjunctival cells by pollen proteases explains conjunctival irritation in proven non-allergic people during the pollen season. One reason why not all people are affected by SNAC syndrome to the same extent could be differences in the concentrations of antiproteases present on the ocular surface.

Eine neue Möglichkeit der Behandlung der Keratokonjunktivitis sicca durch Besprühung mit einer Öl-in-Wasser-Emulsion

ZusammenfassungHintergrundTrotz der vielfältigen Behandlungsmöglichkeiten bei trockenem Auge gibt es bisher keine Therapie, die eine vollständige Heilung dieser Erkrankung ermöglicht. Eine Erweiterung des Therapiespektrums stellt die Besprühung der Augen von Patienten mit Keratokonjunktivitis sicca mit einer Öl-in-Wasser-Emulsion dar. Die Wirkung dieser neuen Behandlungsmethode wurde untersucht.Material und Methode25 Patienten mit trockenem Auge wurden mit einem Aerosol, das Sojabohnenöl, mittelkettige Triglyceride und Phospholipide enthält, besprüht. Die Behandlungszeit betrug jeweils 7 Minuten pro Auge täglich bei insgesamt 14 Anwendungen. Mit einem Fragebogen wurde vor und nach der Behandlung der Symptomscore erhoben. Die Beurteilung des Tränenmeniskus und der lidkantenparallelen Bindehautfalten, die Tränenfilmaufreißzeit, die Fluorescein-Anfärbung der Hornhaut und der Schirmertest in Lokalanästhesie wurden durchgeführt.ResultateEine signifikante Verbesserung der subjektiven Beschwerden, der Tränenfilmaufreißzeit, der Hornhautanfärbung und des Tränenmeniskus wurde beobachtet. Die lidkantenparallelen Falten und der Schirmertest zeigten nach der Behandlung keine signifikante Veränderung.SchlussfolgerungDie neue Methode der Besprühung der Augen von Patienten mit Sicca-Symptomatik mit einer Öl-in-Wasser-Emulsion stellt eine Erweiterung des Therapie-spektrums zur Behandlung dieser Erkrankung des vorderen Augenabschnitts dar.SummaryBackgroundDespite many different therapies of dry eye syndrome a complete healing of this disease is not achievable until now. An enlargement of the therapy spectrum is the spraying therapy for eyes of patients with kerato-conjunctivitis sicca with an oil-in-water-emulsion. The effect of this new treatment was investigated.Materials and methods25 patients with dry eye were sprayed with an aerosol containing soja bean oil, triglycerides of middle chain length and phospholipides. A daily treatment for 7 minutes of each eye was performed for 14 days. Before and after therapy the symptom score was evaluated by a questionnaire. Inspection of the tear meniscus and of the lid-parallel conjunctival folds, the break-up-time, fluorescein-staining of the cornea and the Schirmertest in local anaesthesia were performed.ResultsA significant improvement of subjective symptoms, of the break-up-time, of the corneal staining and of the tear meniscus was observed. The lid-parallel conjunctival folds, and Schirmertest values showed no significant change after treatment.ConclusionsThis new method by spraying the eyes of patients with sicca symptoms with an oil-in-water-emulsion represents an enlargement of the therapies for treatment of this disease of the anterior part of the eye.