Mucosal addressin cell adhesion molecule-1 (MAdCAM-1). Its binding motif for alpha 4 beta 7 and role in experimental colitis

The integrin alpha 4 beta 7 and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) are molecules involved in the normal recirculation of lymphocytes between the blood and the gastrointestinal tract. These molecules may play a complementary and significant role in animal models of colitis. We have investigated the structural interaction between alpha 4 beta 7 and MAdCAM-1. Site-directed mutagenesis studies of the MAdCAM-1 molecule has led to the identification of the amino acid residue (LDT) in the loop between beta strands C and D of the Ig-superfamily-like folds being involved in the adhesive and cell activation functions of MAdCAM-1 with alpha 4 beta 7.     

Paraoxonase (PON1) gene in mice: sequencing, chromosomal localization and developmental expression

PURPOSE: To retrospectively examine the optic disc photographs of a glaucoma population for optic disc haemorrhages, vascular occlusions and vascular abnormalities. METHODS: The optic disc photographs of 906 eyes of glaucoma and suspect glaucoma patients were examined. Optic disc photographs were taken annually, where possible, with the follow-up period varying between 1 and 14 years duration (mean, 2.89). Glaucoma patients are regularly reviewed every 4-6 months and glaucoma suspects every 1-2 years, depending on the ophthalmologist. Low-tension glaucoma patients were reviewed more frequently (mean, every 2.6 months). The results of the findings were compared to a control group of 39 subjects with a mean follow-up period of 7 years, using Fisher's exact test. RESULTS: It was found that during the period under review, 7.4% (n = 67) of eyes had optic disc haemorrhages. The highest frequency of optic disc haemorrhages (37.5%) was found in the low tension glaucoma group (P = 0.0001) followed by 11% of primary open-angle glaucoma eyes (P = 0.03). In the normal group there were three eyes with optic disc haemorrhages and one with a disc collateral, which constitutes 5.1% vascular changes in this sub-group. Of the study eyes 2.8% had central retinal vein occlusions, 1.3% branch vein occlusion, 1.2% disc vessel abnormalities (loops) and 1.1% disc collaterals. Discrete nerve fibre layer haemorrhages and microaneurysms were found in 0.8% and 1.8% of eyes, respectively. CONCLUSIONS: A total of 16.8% of the eyes observed in this study had either disc haemorrhages or vascular changes. The underlying trend of vascular and haemorrhagic changes in glaucoma are demonstrated in this sample, which is in general agreement with previous studies. The high percentage of optic disc haemorrhages in low tension glaucoma is highlighted. The presence of microaneurysms and nerve fibre layer haemorrhages is interesting but of unknown significance.     

Cocaine: mechanism of inhibition of a muscle acetylcholine receptor studied by a laser-pulse photolysis technique

Effects of cocaine on the muscle nicotinic acetylcholine receptor were investigated by using a chemical kinetic technique with a microsecond time resolution. This membrane-bound receptor regulates signal transmission between nerve and muscle cells, initiates muscle contraction, and is inhibited by cocaine, an abused drug. The inhibition mechanism is not well understood because of the lack of chemical kinetic techniques with the appropriate (microsecond) time resolution. Such a technique, utilizing laser-pulse photolysis, was recently developed; by using it the following results were obtained. (i) The apparent cocaine dissociation constant of the closed-channel receptor form is approximately 50 microM. High carbamoylcholine concentration and, therefore, increased concentrations of the open-channel receptor form, decrease receptor affinity for cocaine approximately 6-fold. (ii) The rate of the receptor reaction with cocaine is at least approximately 30-fold slower than the channel-opening rate, resulting in a cocaine-induced decrease in the concentration of open receptor channels without a concomitant decrease in the channel-opening or -closing rates. (iii) The channel-closing rate increases approximately 1.5-fold as the cocaine concentration is increased from 20 to 60 microM but then remains constant as the concentration is increased further. The results are consistent with a mechanism in which cocaine first binds rapidly to a regulatory site of the receptor, which can still form transmembrane channels. Subsequently, a slow step (t1/2 approximately 70 ms) leads to a receptor form that cannot form transmembrane channels, and acetylcholine receptor-mediated signal transmission is, therefore, blocked. Implications for the search for therapeutic agents that alleviate cocaine poisoning are mentioned.     

Comparison of extraction methods and detection systems in the gas chromatographic analysis of volatile carbonyl compounds

High-resolution gas chromatography (HRGC) with electron-capture detection (ECD), nitrogen-phosphorus detection (NPD), flame ionization detection (FID) or with mass spectrometry-selected ion monitoring (MS-SIM) was used in the analysis of volatile carbonyl compounds. Eighteen carbonyl compounds that are typically produced during lipid peroxidation were derivatized quantitatively with pentafluorophenylhydrazine (PFPH) at room temperature, to afford their corresponding water-insoluble hydrazones. These derivatives were extracted into non-polar phases by means of either liquid-liquid extraction (LLE) (hexane) or solid-phase extraction (SPE) on 3 ml C18 octadecyl-bonded phase cartridges. Detection limits of 10(-14) and 10(-12) mol/ml per aldehyde were achieved with the ECD and MS-SIM systems, respectively. The effects of extraction conditions on sensitivity and recovery were determined by performing parallel HRGC-ECD and HRGC-MS-SIM analyses of pentafluorophenylhydrazones of the eighteen compounds under study. Recoveries of 51.4-78.9 +/- 1.2-4.5 and 80.9-98.3 +/- 1.0-3.5% were obtained with LLE and SPE, respectively. The method was applied to the analysis of the volatile carbonyl compounds in various heated vegetable oils (corn, palm or sunflower) and to the analysis of volatile aldehydes in human urine.     

Mutations in purine nucleoside phosphorylase deficiency

PURPOSE: The aim of this study was to investigate the incidence of unexpected malignant uveal melanoma in the age of ultrasound diagnostics and to highlight the reasons for misdiagnosis. PATIENTS AND METHODS: All eyes were surgically removed and histologic examination was performed between 1981 and 1995. The eyes were investigated for the incidence of uveal melanoma, and the history of the unexpected malignant melanoma of the uvea or ciliary body highlighted. RESULTS: 225 (18.7%) eyes with malignant melanoma out of 2583 enucleated eyes were found. Eight (3.6%) of 225 were clinically unexpected. The clinical misdiagnoses were secondary angle closure or open angle glaucoma (6), retinal detachment (5), iritis (1), scleritis (1), cataract (4) and an intraocular mass that was believed to be a metastasis of a colon carcinoma. Seven of eight eyes were blind, and one eye had light perception only. The longest follow up before enucleation was 13 years. On three eyes diagnostic ultrasound was reportedly performed without specific diagnosis of uveal melanoma. Surgery was performed on four eyes for reasons of uncontrollable intraocular pressure or retinal detachment up to five years before enucleation. Histologic diagnoses were 3 epitheloid-type, 2 spindel-type and 3 necrotic melanoma of the uvea. Four eyes showed scleral invasion by tumor cells and one eye an invasion into the episcleral space. CONCLUSIONS: Even today the rate of unexpected uveal melanoma, according to our study is 3.6%. Therefore, all blind eyes without visualisation of the posterior pole should be examined with ultrasound in order to diagnose an uveal melanoma prior to enucleation.     

Intensive care unit prognostic scoring systems to predict death: a cost-effectiveness analysis

OBJECTIVE: To evaluate the cost-effectiveness, using the technique of decision analysis, of withdrawing care from patients in the intensive care unit (ICU) who are predicted to have a high probability of death (>90%) after 48 hrs using a mortality risk estimate based on daily Acute Physiology and Chronic Health Evaluation (APACHE) III scores. MATERIALS AND METHODS: A decision tree model was constructed to compare the cost-effectiveness of two clinical strategies. In the first strategy, patients receive ICU care until they were discharged, died, or had care withdrawn based on subjective clinical criteria. In the second strategy, patients remained in the ICU until they were either discharged, died, or had life-sustaining care withdrawn based on subjective criteria or if they were predicted to have a >90% risk of mortality after 48 hrs by a prognostic scoring system. Transition probabilities were based on a retrospective data analysis of 4,106 noncardiac ICU patients admitted to a tertiary surgical ICU over a 9-yr period. Cost estimates were based on daily Therapeutic Intervention Scoring System (TISS) scores from our database and using published data on the estimated production cost for a TISS point. The sensitivity (16.6%) and specificity (99.6%) of the mortality risk estimate at 48 hrs (using the >90% decision point) based on daily APACHE III scores were derived from published data. RESULTS: In the base case analysis, we assumed that the sensitivity and specificity of the prognostic risk estimate are unchanged when exported to a new environment. Not using a prognostic scoring system as the basis for withdrawing care resulted in a slightly higher survival rate (87.2% vs. 86.85%) at a cost-per-death prevented (CPDP) of $263,700. Since prognostic scoring systems have not been shown to retain the same predictive power when exported to new databases, we chose to explore the effect of varying the specificity of the scoring system on CPDP. Decreasing the specificity from .996 (baseline) to .98 causes the CPDP to drop to $53,300. Changing the specificity to .95 results in a CPDP prevented of $21,700. Using one-way sensitivity analysis, the CPDP is shown to be relatively insensitive to delaying the decision point from ICU day 3 to day 7. Sensitivity analysis also indicates that CPDP increases rapidly with hospital death rate. For a death rate of 30%, the CPDP increases to $768,600 (in the base case, the death rate is 12.8%); when the specificity is decreased to .95, the CPDP drops to $62,100. CONCLUSION: Unless daily mortality risk estimates based on APACHE III can be shown to retain the same level of predictive power in ICUs outside the development database, it is unlikely that the incremental cost-effectiveness gained by using them as the basis to withdraw care is sufficient to justify their use in this manner.