Increase in rat brain glutathione following intracerebroventricular administration of gamma-glutamylcysteine.

The effects of intracerebroventricularly (i.c.v.) administered gamma-glutamylcysteine (gamma-GC) and glutathione (GSH) monoethyl ester, subcutaneously (s.c.) injected L-2-oxo-4-thiazolidinecarboxylic acid (OTC) and intraperitoneally (i.p.) administered cysteine on the concentration of GSH in rat brain were investigated. The brain content of GSH, cysteine and gamma-GC was determined by HPLC with electrochemical detection (gold/mercury electrode) using N-acetylcysteine as internal standard. A dose-dependent increase in the GSH concentration (145-170% of controls) was found in the substantia nigra (SN) and in the rest of the brain stem after injection of gamma-GC, whereas no significant alterations in GSH were observed in the striatum and in the cerebral cortex. High levels of gamma-GC could be detected in the brain tissue after the administration, and the concentration of cysteine did also increase markedly after gamma-GC injection in all brain regions assessed. I.c.v. administration of L-buthionine sulfoximine (L-BSO) reduced the brain concentration of GSH by 50-70% within 24 hr. Injection of gamma-GC 24 hr after L-BSO resulted in an increase in GSH up to control values within 1-3 hr in the SN and the rest of the brain stem, whereas only a slight increase in GSH was observed in the striatum and the cerebral cortex. The concentration of GSH in the striatum and SN did not change after i.p. injection of cysteine, but a slight increase in the GSH concentration in the limbic region was observed. GSH monoethyl ester (i.c.v.) and OTC (s.c.) did not produce any significant increase in the GSH concentration in the brain. When the GSH concentration had been reduced by administration of L-BSO (i.c.v.; 24 hr) subsequent injection of GSH monoethyl ester led to a slight increase in the striatal and limbic GSH levels. These data show that, of the drugs studied, gamma-GC was the most effective in increasing brain GSH. It could thus serve as a valuable tool in future studies regarding metabolism and function of GSH in the brain. The observed difference in the effects of gamma-GC in different brain regions indicate that the brain tissue is not homogeneous with regard to GSH synthesizing capacity.     

Functional analysis of rabbit anti-peptide antibodies which mimic autoantibodies against the beta 1-adrenergic receptor in patients with idiopathic dilated cardiomyopathy.

A synthetic peptide corresponding to the second extracellular loop of the beta 1-adrenergic receptor was used as an antigen for antibody production in three rabbits. Antibodies of high titers were obtained in all rabbits. Only one rabbit yielded antibodies which decreased radioligand binding on the receptor in a similar way to that described for autoantibodies in patients with dilated cardiomyopathy. These antibodies recognized the receptor protein in immunoblots. Epitope mapping indicated that the N-terminal sequence of the loop used as antigen was the target of the major antigen fraction. Incubation of antibodies with C6 glioma cell membranes or inner membranes of E. coli, which express the human beta 1-adrenergic receptor, resulted in a decrease in number of radioligand binding sites. This decrease was dependent on the concentration of antibody and of Mg++ ions. It was not affected by the GTP analog GppNHp or the beta 1 subtype-specific antagonist metoprolol. The agonist, isoproterenol, also induced a decrease but the effects of antibody and agonist were not additive. These results suggest that the antibodies induce a Mg(++)-dependent, 'active', labile conformation of the receptor, independent from coupling to the GTP regulatory protein, but similar to that induced by the agonist isoproterenol. This interpretation was corroborated by the beta 1-adrenergic receptor agonist-like effect of the antibodies on cardiomyocytes in culture.     

Predictive power of various models for longitudinal attachment level change

Abstract Several statistical models that have been suggested in the periodontal literature for describing longitudinal attachment level changes, such as the gradual loss, single-burst, multiple-burst, and random walk models as well as other models introduced in this paper are compared by their power to predict future attachment loss. The data used in this analysis is from 1061 sites of 8 subjects, with moderate to severe periodontal disease, monitored monthly for about a year. This study found that none of the suggested models could significantly outperform the naive mean predictor, which predicts the future attachment level from the past mean. It was also found that no single model, such as the burst, gradual, or random walk, together with measurement error can fully explain the variation in the data. These results indicate that in the course of one year, the attachment level change may not follow the same model. Consequently, a model that fits well to past data cannot be accurately extended to the future.     

In vitro growth characteristics of human odontogenic keratocysts and dentigerous cysts

Using an in vitro system, the growth characteristics and enzyme histochemical properties of 3 odontogenic keratocysts and 3 dentigerous cysts were studied. It was found that the epithelial cells of the keratocysts hut not of the dentigerous cysts grew in vitro. Furthermore, the epithelial-like cells of the keratocysts showed the same activities of acid phosphatase and NADH-diaphorase in vitro as earlier described in vivo. These enzymatic activities were increased in epithelial-like cells close to proliferating fibroblast-like cells, indicating a close relationship between these two cell types. The results are discussed in the light of the known clinical behaviour of the keratocysts and certain conclusions are also drawn concerning the suggested neo plastic potential of the keratocysts.     

Regimen-related toxicity in patients undergoing BMT with total body irradiation using a sweeping beam technique.

In our institution, total body irradiation (TBI) is performed by means of a sweeping beam technique. Toxicity of the procedure was evaluated according to the only grading system designed for high dose chemoradiotherapy. One hundred patients undergoing TBI and conditioned with a standard cyclophosphamide regimen before BMT were evaluated. Regimen-related toxicity was graded according to the Seattle transplantation toxicity system, from 0 to IV (fatal toxicity), in eight organs on days 0, 7, 14, 28 and 100 for lungs. Eighteen patients did not develop any toxicity. Grades III, IV toxicities were uncommon (9%) and were not influenced by dose of TBI, GVHD prophylaxis, disease status and allogenicity although no grade IV toxicity was observed among autologous marrow recipients. However, grade II toxicity was more common in patients receiving allogeneic vs autologous grafts (p < 0.01) because of increased mucosal (p = 0.002) and liver (p = 0.12) toxicities. Renal toxicity was unevaluable. When cumulative toxicity was equal or higher than 4, day 100 survival was worse (p = 0.05). These data confirm the safety of our TBI procedure and the validity of the grading system except for renal toxicity. We suggest that a more aggressive conditioning regimen may be tolerated by patients receiving autologous grafts.     

Long-Term Clinical Outcomes Following Treatment of Actinic Keratosis with Imiquimod 5% Cream

BACKGROUND: The results from four phase III, randomized, vehicle-controlled studies showed that imiquimod 5% cream (imiquimod) was safe and effective in the treatment of actinic keratosis (AK). Patients applied imiquimod or vehicle cream to AK lesions on the face or balding scalp, dosing three times per week or two times per week for 16 weeks. OBJECTIVE: To obtain long-term safety follow-up data and estimate AK recurrence in patients who completely cleared their AK lesions in the treatment area at the 8-week post-treatment visit in the phase III studies. METHODS: One hundred forty-six patients from 30 study centers in the United States were evaluated for clinical evidence of AK, and safety data were collected. RESULTS: After a median follow-up period of 16 months, 24.7% (19 of 77) of the patients administered imiquimod three times per week and 42.6% (23 of 54) of the patients administered imiquimod two times per week had a recurrence of AK (the appearance of at least one AK lesion) in the original treatment area. The median number of AK lesions present was one lesion for both patients receiving imiquimod three times and those receiving imiquimod two times per week compared with a median of six lesions at baseline in the combined three times per week and two times per week phase III studies. There were no long-term safety issues, and the skin quality seen in the imiquimod-treated patients at the end of the phase III studies was maintained. CONCLUSION: One and a half years following treatment, imiquimod continued to provide a long-term clinical benefit in a majority of patients who experienced complete clearance of their AK lesions.     

Cobas-Bact system for identification of members of the family Enterobacteriaceae in 4 h 20 min

The Cobas-Bact (Roche Diagnostics, Basel, Switzerland) new rotor for the identification (ID) in 4 h 20 min of 33 members of the family Enterobacteriaceae to genus and species level was evaluated by testing 444 strains of which 398 belonged to common species and 46 belonged to rare species of Enterobacteriaceae. Each strain was identified by the API 20E system (Analytab Products, Plainview, N.Y.), and additional discriminating tests were set up if necessary. Only first-choice ID were considered in this study and were classified either as high-confidence ID (normalized likelihood, greater than or equal to 80%) or as low-confidence ID (normalized likelihood, less than 80%) requiring additional tests for confirmation. The data were analyzed by two versions of Cobas-Bact software. With the first version of the software (SW8446), the overall accuracy of Cobas-Bact was 95.5% (424 correct ID of 444). When restricted to high-confidence ID it rose to 99.4% (350 of 352) for the common species and 96.9% (31 of 32) for the rare species. Only three strains of the high-confidence group were misidentified. Sixty ID were considered unacceptable because of their low confidence. Using the first software version (SW8446) they represented 12% (46 of 398) of the common species (17 typical strains, 10 Shigella species, 10 inactive Escherichia coli strains, and 9 rare biotypes) and 30% (14 of 46) of the rare species. The same data analyzed by the new version (SW8524) of the Cobas-Bact software resulted in an overall accuracy of 93.9% (417 correct ID). The number of high-confidence ID rose to 401, of which 392 (97.7%) were accurate. The decrease in low-confidence ID (43 versus 60) was mainly due to the Shigella species. In conclusion the accuracy of Cobas-Bact identification system was very good when restricted to high-confidence ID. The Cobas-Bact performance for rare species ID was poorer, but the small number of strains tested does not allow definitive conclusions.