Sites of volatile anesthetic action on kainate (Glutamate receptor 6) receptors

Molecular mechanisms of anesthetic action on neurotransmitter receptors are poorly understood. The major excitatory neurotransmitter in the central nervous system is glutamate, and recent studies found that volatile anesthetics inhibit the function of the alpha-amino-3-hydroxyisoxazolepropionic acid subtype of glutamate receptors (e.g. glutamate receptor 3 (GluR3)), but enhance kainate (GluR6) receptor function. We used this dissimilar pharmacology to identify sites of anesthetic action on the kainate GluR6 receptor by constructing chimeric GluR3/GluR6 receptors. Results with chimeric receptors implicated a transmembrane region (TM4) of GluR6 in the action of halothane. Site-directed mutagenesis subsequently showed that a specific amino acid, glycine 819 in TM4, is important for enhancement of receptor function by halothane (0. 2-2 mM). Mutations of Gly-819 also markedly decreased the response to isoflurane (0.2-2 mM), enflurane (0.2-2 mM), and 1-chloro-1,2, 2-trifluorocyclobutane (0.2-2 mM). The nonanesthetics 1, 2-dichlorohexafluorocyclobutane and 2,3-dichlorooctafluorobutane had no effect on the functions of either wild-type GluR6 or receptors mutated at Gly-819. Ethanol and pentobarbital inhibited the function of both wild-type and mutant receptors. These results suggest that a specific amino acid, Gly-819, is critical for the action of volatile anesthetics, but not of ethanol or pentobarbital, on the GluR6 receptor.     

Design of low cost glaucoma screening

In 1991 the Netherlands Glaucoma Patient Association organized a glaucoma screening survey. This survey was designed to evaluate the effectiveness of a low cost screening setting. During a screening period of 8 days, 1259 subjects over the age of 49 years were examined by a team of non-ophthalmologically trained students. The following screening methods were used: visual field analysis (Henson CFS3000 perimeter), retinal nerve fiber layer photography (Canon non-mydriatic camera), intraocular pressure measurement (Pulsair non-contact tonometer) and determination of the peripheral anterior chamber depth (slitlamp biomicroscope, the van Herick method). In a later stage, subjects with glaucomatous abnormalities in the visual field and/or the photograph were re-examined by a glaucoma specialist using applanation tonometry, gonioscopy, ophthalmoscopy and Humphrey 30-2 visual field analysis. The time taken to conduct the individual screening tests in a subject varied from 1 to 5 min: perimetry took 5 min, photography 2 min, tonometry 3 min and angle-width determination 1 min. Fifty-six (4.4%) subjects showed glaucomatous defects in perimetry and/or photography. Thirty-seven could be re-examined and glaucoma was diagnosed in 16 subjects. Visual field defects and glaucomatous abnormalities in the photograph were confirmed by Humphrey perimetry in 72.7% and 35.7% respectively. Sixty-seven (5.3%) subjects had an intraocular pressure above 21 mm Hg, while no cases of angle closure glaucoma were found in this population. The costs of this screening setting were estimated at F1. 48,60 per screen. A future low cost screening survey might be limited to non-contact tonometry and visual field analysis with the Henson CFS3000 perimeter or a similar device, using suprathreshold testing with a limited number of points. Screening might be performed by non-medically trained employees. The costs of such a screening program may be estimated at F1. 16,- per screen and F1. 1.989,- per glaucoma case using a mobile screening unit (addendum).     

Use of urinary cotinine and questionnaires in the evaluation of infant exposure to tobacco smoke in epidemiologic studies

In this study we have investigated whether proteoglycans (aggrecan) are modified by nonenzymatic glycation as in collagen. Purified human aggrecan from osteoarthritic and normal human knee articular cartilage was assayed for pentosidine, a cross-link formed by nonenzymatic glycation, using reverse-phase HPLC. In addition, an in vitro study was done by incubation of purified bovine nasal cartilage aggrecan with ribose. Pentosidine was found in all the purified human aggrecan samples. 2-3% of the total articular cartilage pentosidine was found in aggrecan. Purified link protein also contained penosidine. The in vitro study led to pentosidine formation, but did not appear to increase the molecular size of the aggrecan suggesting that pentosidine was creating intramolecular cross-links. Similar amounts of glycation were found in osteoarthritic and normal cartilage. Like collagen, aggrecan and link proteins are crosslinked by nonenzymatic glycation in normal and osteoarthritic cartilage. Crosslinking could be reproduced, in vitro, by incubating aggrecan with ribose.     

Diltiazem-associated thrombocytopenia

Nine days after starting diltiazem therapy for new-onset atrial fibrillation, a patient's platelet count had diminished to 61 x 10(3)/mm3, and 2 weeks later the nadir was reached at 23 x 10(3)/mm3. No hypersensitivity reaction otherwise was noted, and other hematologic values were unaffected. The patient's platelet counts were not affected by platelet transfusions. Bone marrow examination showed normal to increased numbers of megakaryocytes, suggesting peripheral destruction. After discontinuing diltiazem and administering high-dose immunoglobulin infusions, the platelet count returned to normal. This case suggests an immune-mediated cause for thrombocytopenia after exposure to diltiazem.     

Renal and vascular effects of chronic endothelin receptor antagonism in malignant hypertensive rats

The effect of the combined ETA/ETB endothelin receptor antagonist bosentan on blood pressure, vascular hypertrophy, and pathologic renal changes was investigated in a model of malignant hypertension, severe vascular hypertrophy, and enhanced vascular expression of endothelin-1, the deoxycorticosterone acetate (DOCA), and salt-treated spontaneously hypertensive rat (SHR). DOCA-salt treated SHR received 100 mg bosentan per kilogram weight per day mixed with their food. Systolic blood pressure of untreated DOCA-salt SHR rose to 241 +/- 1.5 mm Hg, whereas that of bosentan-treated rats rose to 221 +/- 5.1 mm Hg (P < .01). Cardiac and conduit artery mass were not affected by treatment. Small arteries from the coronary, renal, and mesenteric circulations showed a smaller media width and cross-sectional area of the media in rats treated with bosentan than in untreated rats. The kidneys showed the presence of fibrinoid necrosis in a high percentage of afferent arterioles and glomeruli of untreated DOCA-SHR. Some kidneys of treated rats exhibited less severe vascular hypertrophy and lesser extent of vascular or glomerular fibrinoid necrosis, but the renal injury score of bosentan-treated DOCA-SHR was only at the limit of significance from that of untreated rats (P = .06). These results suggest a role for endothelin-1 in blood pressure elevation and the severe vascular hypertrophy of small arteries of the coronary, renal, and mesenteric vasculature, but not of the heart or larger conduit vessels in the malignant hypertension that SHR develop after treatment with DOCA and salt. Although some bosentan-treated rats showed fewer renal lesions, a significant effect on renal pathology could not be unambiguously demonstrated. Further studies will be necessary to determine whether endothelin antagonists may indeed offer some degree of renal protection and have therapeutic potential in severe or malignant hypertension.     

Heat-Shocked Lactobacilli for Acceleration of Cheddar Cheese Ripening

The aim of this study was to establish adequate conditions for heat-shocking cells of lactobacilli, to sufficiently suppress lactic acid production without damaging the proteolytic enzyme system important for cheese maturation. Three temperatures, 65, 67 and 70°C were tested, for 22 sec. The best combination for maximum retardation of lactic acid production and minimum damage to the proteolytic system was obtained by treating cells at 67°C for 22 sec. Following such treatment, lactic acid production was retarded by 24 hr, while the proteolytic enzyme system remained scarcely unchanged.