Adipokines as atherothrombotic risk factors in obese subjects: Associations with haemostatic markers and common carotid wall thickness.

BACKGROUND AND AIMS: Some crucial associations between obesity-related altered adipokine levels and the main factors of atherosclerotic, atherothrombotic processes are not fully known. We analysed the relationships of classic adipokines, namely leptin, resistin, adiponectin, tumour necrosis factor-alpha (TNF-α), interleukin 6 (IL-6) with the markers of platelet activation, including mean platelet volume (MPV), platelet surface/soluble P-selectin, platelet-derived microparticles (PMPs), the parameters of coagulation abnormalities and common carotid intima-media thickness (IMT) in obese patients with or without atherosclerotic comorbidities in comparison to age- and sex-matched controls. METHODS AND RESULTS: We enrolled 154 obese individuals, including 98 suffering from atherosclerotic concomitant conditions, 56 free of atherosclerotic comorbidities and 62 healthy controls. Plasma levels of leptin, resistin, adiponectin, TNF-α, IL-6, soluble P-selectin, and plasminogen activator inhibitor-1 antigen (PAI-1 ag) were analysed by ELISA. Platelet surface P-selectin and PMPs were measured by flow cytometry. IMT was detected by ultrasonography. Adipokines were closely associated with markers of platelet hyperactivity, hypercoagulability, hypofibrinolysis and IMT. Significant independent associations were found between leptin and platelet count (p < 0.0001), MPV (p = 0.019), PMPs (p < 0.0001), fibrinogen (p = 0.001), factor VIII (FVIII) activity (p = 0.035); adiponectin and PAI-1 ag (p = 0.035); resistin and soluble P-selectin (p = 0.002); TNF-α and PAI-1 ag (p < 0.0001); and IL-6 and fibrinogen (p = 0.011). Finally, leptin (p = 0.0005), adiponectin (p = 0.019), IL-6 (p = 0.001), MPV (p = 0.0003), PMP (p = 0.008), and FVIII activity (p = 0.043) were independent predictors of IMT. CONCLUSION: Overall, we suggest that in obese subjects altered adipokine levels play a key role in common carotid atherosclerosis both directly and through haemostatic parameters.

Clearance of autophagy-associated dying retinal pigment epithelial cells - a possible source for inflammation in age-related macular degeneration.

Retinal pigment epithelial (RPE) cells can undergo different forms of cell death, including autophagy-associated cell death during age-related macular degeneration (AMD). Failure of macrophages or dendritic cells (DCs) to engulf the different dying cells in the retina may result in the accumulation of debris and progression of AMD. ARPE-19 and primary human RPE cells undergo autophagy-associated cell death upon serum depletion and oxidative stress induced by hydrogen peroxide (H2O2). Autophagy was revealed by elevated light-chain-3 II (LC3-II) expression and electron microscopy, while autophagic flux was confirmed by blocking the autophago-lysosomal fusion using chloroquine (CQ) in these cells. The autophagy-associated dying RPE cells were engulfed by human macrophages, DCs and living RPE cells in an increasing and time-dependent manner. Inhibition of autophagy by 3-methyladenine (3-MA) decreased the engulfment of the autophagy-associated dying cells by macrophages, whereas sorting out the GFP-LC3-positive/autophagic cell population or treatment by the glucocorticoid triamcinolone (TC) enhanced it. Increased amounts of IL-6 and IL-8 were released when autophagy-associated dying RPEs were engulfed by macrophages. Our data suggest that cells undergoing autophagy-associated cell death engage in clearance mechanisms guided by professional and non-professional phagocytes, which is accompanied by inflammation as part of an in vitro modeling of AMD pathogenesis.

Structure-activity relationships of vanilloid receptor agonists for arteriolar TRPV1.

BACKGROUND AND PURPOSE: The transient receptor potential vanilloid 1 (TRPV1) plays a role in the activation of sensory neurons by various painful stimuli and is a therapeutic target. However, functional TRPV1 that affect microvascular diameter are also expressed in peripheral arteries and we attempted to characterize this receptor. EXPERIMENTAL APPROACH: Sensory TRPV1 activation was measured in rats by use of an eye wiping assay. Arteriolar TRPV1-mediated smooth muscle specific responses (arteriolar diameter, changes in intracellular Ca(2+)) were determined in isolated, pressurized skeletal muscle arterioles obtained from the rat and wild-type or TRPV1(-/-) mice and in canine isolated smooth muscle cells. The vascular pharmacology of the TRPV1 agonists (potency, efficacy, kinetics of action and receptor desensitization) was determined in rat isolated skeletal muscle arteries. KEY RESULTS: Capsaicin evoked a constrictor response in isolated arteries similar to that mediated by noradrenaline, this was absent in arteries from TRPV1 knockout mice and competitively inhibited by TRPV1 antagonist AMG9810. Capsaicin increased intracellular Ca(2+) in the arteriolar wall and in isolated smooth muscle cells. The TRPV1 agonists evoked similar vascular constrictions (MSK-195 and JYL-79) or were without effect (resiniferatoxin and JYL-273), although all increased the number of responses (sensory activation) in the eye wiping assay. Maximal doses of all agonists induced complete desensitization (tachyphylaxis) of arteriolar TRPV1 (with the exception of capsaicin). Responses to the partial agonist JYL-1511 suggested 10% TRPV1 activation is sufficient to evoke vascular tachyphylaxis without sensory activation. CONCLUSIONS AND IMPLICATIONS: Arteriolar TRPV1 have different pharmacological properties from those located on sensory neurons in the rat.

Antiproliferative effect of 4-thiouridylate on OCM-1 uveal melanoma cells.

PURPOSE: Brachytherapy is a well-established, effective treatment for uveal melanoma with a failure rate of 15%. The fatal consequence of unsuccessful treatments offers reason for improvement of the method. The authors propose using an apoptosis inducing agent locally, concomitantly with the well-established therapy, to sensitize the tumor cells. The authors propose a new nontoxic moderately active apoptosis inducing agent, 4-thio-uridylate (s4UMP), for this purpose. METHODS: OCM-1 uveal melanoma cells were treated with various concentrations of s4UMP and its effect was monitored by measuring the cell viability (MTT assay). The following apoptosis detecting methods were performed to reveal the mechanism of decreased cell viability: light microscopy, DNA fragmentation assay, determination of caspase 9 activity, and FACS analysis. RESULTS: The viability of uveal melanoma cells was decreased by 32%, 40%, and 9% after 24, 48, and 72 hours of treatment with 10 microg/mL (30 microM) s4UMP. The effect was not dose dependent; it rather followed a saturation-type inhibition and the cells at lower drug concentration recovered after 72 hours. Characteristic apoptotic cell morphology and DNA fragmentation was detected in treated cells. The caspase-9 was activated upon treatment showing maximal activity at 48 hours suggesting the induction of apoptosis. The annexin binding activity further verified the apoptogenic activity of s4UMP. CONCLUSIONS: Uveal melanoma, more than other solid tumors, is resistant to most of the chemotherapeutic protocols as indicated by the high mortality rate of metastatic disease. The authors showed that s4UMP, a naturally occurring nucleotide, could induce apoptosis in uveal melanoma cells, suggesting a potential supplementary therapeutic application of the compound.

Antiproliferative effect of 4-thiouridylate on OCM-1 uveal melanoma cells.

PURPOSE: Brachytherapy is a well-established, effective treatment for uveal melanoma with a failure rate of 15%. The fatal consequence of unsuccessful treatments offers reason for improvement of the method. The authors propose using an apoptosis inducing agent locally, concomitantly with the well-established therapy, to sensitize the tumor cells. The authors propose a new nontoxic moderately active apoptosis inducing agent, 4-thio-uridylate (s4UMP), for this purpose. METHODS: OCM-1 uveal melanoma cells were treated with various concentrations of s4UMP and its effect was monitored by measuring the cell viability (MTT assay). The following apoptosis detecting methods were performed to reveal the mechanism of decreased cell viability: light microscopy, DNA fragmentation assay, determination of caspase 9 activity, and FACS analysis. RESULTS: The viability of uveal melanoma cells was decreased by 32%, 40%, and 9% after 24, 48, and 72 hours of treatment with 10 g/mL (30 M) s4UMP. The effect was not dose dependent; it rather followed a saturation-type inhibition and the cells at lower drug concentration recovered after 72 hours. Characteristic apoptotic cell morphology and DNA fragmentation was detected in treated cells. The caspase-9 was activated upon treatment showing maximal activity at 48 hours suggesting the induction of apoptosis. The annexin binding activity further verified the apoptogenic activity of s4UMP. CONCLUSIONS: Uveal melanoma, more than other solid tumors, is resistant to most of the chemotherapeutic protocols as indicated by the high mortality rate of metastatic disease. The authors showed that s4UMP, a naturally occurring nucleotide, could induce apoptosis in uveal melanoma cells, suggesting a potential supplementary therapeutic application of the compound.

[Corneal transplantation in Hungary. Data of the National Keratoplasty Registry 1992-1996]. / Szaruhártya-átültetés Magyarországon. Az Országos Keratoplasztika Regiszter 1992 és 1996 közötti idöszakra vonatkozó adatai.

Upon the request of the National Transplantation Committee a nationwide keratoplasty register is functioning at the Department of Ophtalmology, University Medical School of Debrecen. Since then at the end of each year we collected data from the University Departments and Hospitals performing corneal transplantation concerning their keratoplasties and related activities. According to the acquired data the number of corneal transplantations performed in Hungary was 299 in 1992, 298 in 1993, 320 in 1994, 426 in 1995, and 479 in 1996. Keratoplasties in Hungary were performed in 6 University Departments and in 13 Hospitals, all together in 19 institutions in the past five years. The overwhelming majority of corneal transplantations (in 1995 95.8% and in 1996 96.4%) were penetrating keratoplasties. The ratios of lamellar keratoplasties in 1995 and in 1996 were 3.0% and 2.5%, respectively. The ratios of sclerokeratoplasties were slightly above 1% in both years. The distributions of implanted fresh and preserved (eye bank) corneas were around 50-50% in these two years so, that in 1995 a somewhat more and in 1996 somewhat less preserved corneas were used. This ratio for preserved corneas was less than 20% in the period between 1992 and 1994. The law regulating the procurement and the transplantation of corneas is based on presumed consent and explicit objection which is the better type of regulation for the performance of keratoplasties in large numbers. The number of corneal transplantations in Hungary in the past years was close to the figures of industrial countries (England, Germany) calculated for similar populations, inspite of the fact that financing was in many respects unacceptable.

Changing techniques and indications for lamellar keratoplasty.

In recent decades, the number of lamellar keratoplasties (LKs) being performed worldwide has been gradually decreasing. However, new technical procedures have been developed and microsurgeons must consider modified indications for lamellar grafting. In the period 1946-1995, in the Department of Ophthalmology University Medical School of Debrecen, Hungary 3889 consecutive keratoplasties were performed of which 608 were LKs. In the period 1946-1960 special surgical material (biosutures) was used without operating microscopes and the leading indication for LK was corneal scar (leukomas). Between 1961 and 1979 with the development of microsurgical procedures (10/0 nylon sutures, operating microscope), the LK technique became more advanced. In the period 1979-1995 precision lamellar grafting could be performed with the widely available improved modern techniques. Changing indications for LK were noted in the second period and this tendency has subsequently continued. Currently the main indications for LK are "immediate" keratoplasty after chemical burns, and sclerokeratoplasty against recurrent pterygia. Their observations lead the authors to conclude that the possible reasons for the changing indications of LK are the new technical procedures, the increasing knowledge, the better immunological tests and the improved postoperative therapy.

New indications for and technics in vitreoretinal surgery.

Vitreoretinal surgery has become a widely used ophthalmic surgical procedure since the late sixties. However, it remained an intensively developing field of surgical interventions, and indications still become wider and wider. The procedure starts with the removal of vitreous body, blood or inflammatory material from the vitreous cavity. After this, the given retinal disease, such as retinal detachment, macular hole, proliferative diabetic retinopathy, intraocular foreign body, etc. can be properly treated in situ. For that purpose a number of fine intraocular instruments, fluids, gases can be used. While the anatomical success rate can be up to 95%, the final visual outcome, due to the fact that the retina is not rarely seriously damaged, is less enthusiastic. However, a great number of previously untreatable vitreoretinal diseases can be managed in this way, nowadays.

Effects of chronic alcohol ingestion on myocardial lipid and fatty acid composition in adult turkeys.

STUDY OBJECTIVE: The aim was determine whether alcohol induced left ventricular dilatation and myocardial lesions in turkeys are associated with changes in the phospholipid and fatty acid compositions of the cardiac tissue. DESIGN: Triglyceride, phospholipid, and cholesterol concentrations and the fatty acid compositions of phospholipids and triglycerides were measured in the myocardium of control and alcohol fed turkeys. Tissue samples were taken from the anteroapical wall of the left ventricle. SUBJECTS: Seven week old domestic Nicholas turkeys were used (10 controls and 10 ethanol fed); the alcohol, comprising more than 20% of the dietary energy, was given in the drinking water for 16 weeks. MEASUREMENTS AND MAIN RESULTS: Left ventricular function was assessed by echocardiography at the end of the treatment. Different phospholipid fractions were isolated by two dimensional thin layer chromatography, and individual fatty acids were identified by gas chromatography. Non-invasive evaluation of the cardiac function revealed a dilatation of the left ventricle and an impaired contractile function in the alcoholic birds. Increased amounts of triglyceride, phosphatidylinositol, sphingomyelin, lysophosphatidylcholine, and lysophosphatidylethanolamine were found in the left ventricle of the alcoholic turkeys. Examination of the fatty acid compositions of different phospholipids and the triglyceride fraction revealed an increase in the saturated/unsaturated ratio following ethanol treatment. In the phospholipid fractions of the alcoholic birds the arachidonic acid (20:4n-6) content was significantly decreased as were the linolic (18:1) and linoleic (18:2n-6) acid contents in the triglycerides. CONCLUSIONS: These alterations in the myocardial phospholipid and fatty acid compositions may contribute to the electrophysiological and functional derangements of the left ventricle in alcoholic cardiomyopathy.

[Unilateral infiltration of the optic papilla in chronic lymphatic leukemia]. / Einseitige Papilleninfiltration bei chronisch lymphatischer Leukämie.

This paper reports the clinical history of a 72-year-old woman in whom unilateral papilledema was the only organic manifestation of chronic lymphatic leukemia. In the light of the laboratory parameters direct leukemic infiltration of the optic disk was diagnosed by fundus examination and fluorescein angiography. Combined cytostatic therapy bed to a considerable improvement in the patient's clinical condition.