Muscarinic cholinoceptor-stimulated phosphatidyl inositol pathway in corneal epithelial and endothelial cells

Background Muscarinic cholinoceptors are distributed widely in both the central and peripheral nervous system. The presence of muscarinic cholinoceptors in corneal tissue is well established. Previous reports have shown that corneal muscarinic cholinoceptors are of the m2 or m4 subtype. However, recent studies have indicated the presence of the m5 muscarinic cholinoceptor subtype in human corneal epithelium and endothelium. The aim of the study was to confirm the presence of the m5 cholinoceptor subtype in bovine corneal epithelium and endothelium and the activation of phosphatidyl inositol pathway by its stimulation. Methods Muscarinic m5 cholinoceptor sites, phosphatidyl inositol 4,5-biphosphate, inositol 1,4,5-triphosphate and protein kinase C, were studied using immunocytochemistry and immunofluorescence. Activation of protein kinase C after stimulation of the m5 muscarinic cholinoceptor subtype was measured using the HTS protein kinase C assay kit. Results Immunocytochemistry/immunofluorescence revealed the presence of the m5 muscarinic cholinoceptor subtype, phosphatidyl inositol 4,5-biphosphate and protein kinase C in bovine corneal epithelial and endothelial cells. In bovine corneal epithelium and endothelium, protein kinase C activity was stimulated by acetylcholine in a dose-dependent manner (P<0.0001). Conclusions Our findings indicate that acetylcholine-induced stimulation of muscarinic m5 cholinoceptors activates the phosphatidyl inositol pathway in corneal epithelial and endothelial cells, resulting in increased protein kinase C activity. Further work will be needed to clear the physiologic role of this signaling pathway in corneal epithelium and endothelium. The authors have no proprietary interest in any of the products used in the study.     

Trabecular aspiration in pseudoexfoliative glaucoma--surgery to primarily reduce intraocular pressure

BACKGROUND: Trabecular aspiration has been discussed during the past few years as a new surgical method in the treatment of pseudoexfoliative glaucoma. In this procedure PEX-material, pigment and detritus are aspirated from the trabecular meshwork. Trabecular aspiration has been evaluated mainly in combination with cataract extraction. This study reports on our first experiences concerning the IOP-reducing effect of trabecular aspiration as a primary surgical method in the management of pseudoexfoliative glaucoma. MATERIALS: 17 eyes of 14 patients (7 m, 7 f; 12 OD, 5 OS; age 71 +/- 6 years) with pseudoexfoliative glaucoma were included in this study and operated on by standardised trabecular aspiration (vacuum max. 200 mm Hg, 180 - 270 degrees, 5 min). Therapy success was defined as an IOP reduction by more than 20 % and less than 21 mm Hg. RESULTS: Therapy success was 82 % (14 out of 17) on the first postoperative day, 50 % after 30 days (8 out of 16) and 23 % after 180 days (3 out of 13). IOP was 26.8 +/- 8.5 mm Hg before surgical intervention, 18.1 +/- 11.4 mm Hg after 1 day, 19.1 +/- 7.9 mm Hg after 30 days and 19.2 +/- 5.2 mm Hg after 180 days. Mean quantity of antiglaucomatous eye drops application was 3.1 +/- 0.9 preoperatively, 0.9 +/- 1.6 after 1 day, 0.8 +/- 1.2 after 30 days and 1.0 +/- 1.3 after 180 days. CONCLUSIONS: Trabecular aspiration achieves a good short-term effect in the reduction of IOP in patients with pseudoexfoliative glaucoma. However, this effect was limited to a few weeks in most patients. Trabecular aspiration as a primary surgical method in the management of pseudoexfoliative glaucoma does not appear to be suitable for long-term IOP reduction.     

Local application of vancomycin for prophylaxis of graft infection: Release of vancomycin from antibiotic-bonded dacron grafts,toxicity in endothelial cell culture,and efficacy against graft infection in an animal model

Methicillin-resistant strains ofStaphylococcus epidermidis cause an increasing number of prosthetic infections. This prompted us to test the uptake of vancomycin in various graft materials in vitro, its influence on graft healing, and its efficacy against graft infection in pigs. Incubation of six different Dacron graft materials in a vancomycin solution (20 gm/L) was performed. Grafts were then placed in plasma, and samples were taken over 72 hours to determine vancomycin levels. Release of vancomycin ranged from 775 µg/cm² to 3691 µg/cm² after 1 hour of incubation. Gelatin-covered grafts increased release of vancomycin fourfold when incubation time was extended to 24 hours; uncovered grafts or the collagen-covered graft did not. Graft healing was not complicated when a vancomycin-bonded, gelatin-impregnated Dacron graft was implanted to replace the common femoral artery in pigs. Four weeks after implantation, histologic examination revealed normal development of neointima and perigraft scar tissue in the vancomycin-treated (n=4) and untreated (n=5) grafts. To test the efficacy of local vancomycin against graft infection, grafts were implanted in the groin of pigs and contaminated with 2 × 10⁷ colony-forming units ofStaphylococcus aureus. Four weeks after implantation, all grafts were infected in the untreated group (n=6), with abscess, nonincorporated graft, and detection ofS. aureus from the graft. In the treatment group (n=6) vancomycin was added to the contaminated grafts. As a carrier for the vancomycin, we used a resorbable gelatin-glycerol foam. All grafts healed without infection. The difference between the treated and untreated groups is statistically significant (p<0.05). We conclude that it may be effective to prevent graft infection with local application of vancomycin if an in situ replacement of infected graft (infected by gram-positive bacteria) is necessary or if there is a high risk of infection by methicillin-resistant staphylococci.     

The effects of harmine on the transmembrane action potentials of guinea-pig left atria as compared to those of quinidine

Summary The actions of quinidine (7.7 and 25.5 M) and harmine (10,33 and 66 M) on the action potential of isolated guinea-pig atria were studied. Both substances prolonged repolarization time dose-dependently. The depolarization velocity was considerably reduced by quinidine, while it was comparatively little affected by harmine. Antiarrhythmic compounds of the harmine type are rare.The effects on the action potential explain the differences between quinidine and harmine with regard to effects on the ECG and antagonism of aconitine-induced arrhythmia.