Chronopharmacological study of bunazosin hydrochloride in healthy subjects

Bunazosin, an alpha-1 adrenoreceptor blocking agent, was given orally to six healthy subjects in the morning or in the evening in order to examine the time-dependent variations in the pharmacological effects of the drug. The study was carried out on four occasions in a placebo controlled cross-over design. No significant change was observed in blood pressure either after the morning or the evening dosage. Finger skin blood flow (FSBF) assessed by laser Doppler flowmetry increased significantly after the morning dosage, but not after the evening trial. The plasma concentration of bunazosin in the morning in general had a tendency to be greater than that in the evening. A significant correlation was observed between the plasma concentrations and the increments in the FSBF. These results indicate that the pharmacological effects of bunazosin are greater following morning dosage. The present study supports the concept that the time-dependent differences in the effect of bunazosin are, at least, caused by the time-dependent changes in plasma drug concentration.     

Interaction of bile acids, phospholipids, cholesterol and triglyceride with dietary fibers in the small intestine of rats

Certain dietary fibers have been reported to lower plasma cholesterol by binding bile acids and reducing their recycling through the enterohepatic circulation. In addition, certain fibers may delay the digestion and absorption of fat. In the present study, the interaction of bile acids with guar gum (GG), konjac mannan (KM) and chitosan (CH) was determined. Rats were fed during a 20-min period a test meal containing either 5% cellulose (CE), GG, KM or CH and also containing 14C-labeled triolein and 3H-labeled cholesterol. The group fed CE served as control, since CE does not bind bile acids or phospholipids in vivo. Two hours after presentation of the test meal, rats were killed and the stomach and small and large intestine removed. All four groups ate the same amount of the test meal, about 1.9 g. The aqueous phase of the small intestinal contents was separated by ultracentrifugation, and the amount (mumol) of bile acids and phospholipids in the total intestinal contents and in the aqueous phase was estimated. The ratio of bile acids in the aqueous phase to that in total intestinal contents was significantly higher in the GG and KM groups and significantly lower in the CH group than that in the CE group, demonstrating that the bile acids are bound or trapped by each of these fiber sources. Only CH appeared to bind phospholipids, reducing the proportion in the aqueous phase compared to that in the CE group.(ABSTRACT TRUNCATED AT 250 WORDS)     

CD8+ T cell subsets of cytotoxic T lymphocytes induced by Epstein-Barr virus infection in infectious mononucleosis

Mononuclear peripheral blood lymphocytes (PBL) from patient with infectious mononucleosis (IM) were tested in a 51Cr-release assay for cytotoxicity against autologous and allogeneic lymphoblastoid cell line (LCL), or Epstein-Barr virus (EBV)-genome positive and negative cell line. In acute phase, PBL lyse an autologous LCL as well as allogeneic LCL (Wa cells). High levels of cytotoxicity were observed in the combinations between effector and target cells sharing HLA-Class 1 product. EBV-genome positive Daudi and Raji cells which lack HLA-Class 1 antigen and have mismactched HLA-Class 1 antigen, respectively showed resistance to killing. EBV-genome negative tumor cells except NK sensitive K562 cells were not killed by IM lymphocytes. However, the IM lymphocytes without atypical form in convalescent phase failed to show killing activity against autologous and allogeneic LCL. These findings suggest that cell surface membrane antigen structure on EBV-infected LCL may be able to explain the recognition and triggering of lysis of target cells by HLA-Class 1 restricted cytotoxic T cells (CTL) from acute IM. Phenotypic analysis of PBL with atypical form from IM was made by two-color flow cytometry. The data demonstrate that CD8+ T cells quantitatively represent the major population of lymphocytes expanded during acute IM. Furthermore, approximately 70% of these CD8+ T cells express HLA-DR on these surface, suggesting that they have undergone activation. However, IL 2R (CD25 antigen) expression was not significantly elevated on activated T cells. The salient profile on cytofluorographs of an acute IM was the increased number of CD3+CD19-, CD8+CD11b-, CD8+CD28+ and CD8+S6F1+ cells. However, CD3-CD19+, CD8+CD11b+, CD8+S6F1-, CD4+Leu8- and CD25+HLA-DR+ antigens were little expressed. Increased number of CD8+CD11b-, CD8+CD28+ and CD8+S6F1+ cells, which are regarded as CTL were reduced according to the improvement of the clinical symptoms and laboratory findings. These results together with HLA typing analysis suggested a possibility HLA-Class 1 restriction of the CTL with surface phenotype of CD8+CD11b-, CD8+CD28+, and CD8+S6F1+.     

Human autoantibodies against the 230-kD bullous pemphigoid antigen (BPAG1) bind only to the intracellular domain of the hemidesmosome, whereas those against the 180-kD bullous pemphigoid antigen (BPAG2) bind along the plasma membrane of the hemidesmosome in normal human and swine skin.

Bullous pemphigoid (BP) is a blistering skin disease in which autoantibodies develop to hemidesmosomal components of the epidermal basement membrane zone, including two major antigenic proteins of the 230-kD antigen (BPAG1) and the 180-kD antigen (BPAG2). The present study demonstrated the precise ultrastructural localization of the epitopes for autoantibodies against BPAG1 and BPAG2 in normal skin. Autoantibodies against either BPAG1 or BPAG2 were affinity-purified using nitrocellulose membrane, which was blotted with SDS-PAGE-fractionated antigens from human epidermal extract as the immunoabsorbent. Postembedding, immunogold electron microscopy was performed after skin was processed by rapid freezing and freeze substitution fixation without chemical fixatives. Purified autoantibodies against BPAG1 bound only to the intracellular domain of the hemidesmosome, and 80% of the gold labeling was within 40-140 nm from the plasma membrane (mean distance 91 nm inside). In contrast, the autoantibodies against BPAG2 bound along the plasma membrane of the hemidesmosome, and 80% of the gold labeling was within 10 nm outside to 50 nm inside the cells (mean distance 12 nm inside). These results suggest that the autoantibodies against BPAG1 and BPAG2 react with the epitopes localizing in distinct regions of the hemidesmosome complex, and may play different roles in the blister formation in patients with BP.     

Endoscopic diagnosis and treatment of a case of respiratory papillomatosis

Respiratory papilloma is a rare lesion that arises in the larynx, trachea and bronchus. We describe a patient with laryngeal papilloma that spread to the trachea and which was effectively treated by Nd-YAG laser. Two years after the initial treatments of the laryngeal and tracheal papillomas, a recurrent lesion (solitary papilloma) was observed on the membranous portion of the trachea. We examined the recurrent lesion by bronchoscopy including bronchoscopic ultrasound (US), helical computed tomography (CT) and tracheal biopsies. Respiratory papillomatosis sometimes shows either malignant transformation or invasion to tracheal wall without displaying cytohistological atypia. Therefore, we concluded that bronchoscopic US and helical CT were useful for deciding on therapeutic strategy in cases of recurrence of tracheal papilloma.     

Population Pharmacokinetics of Digoxin in Duchenne Muscular Dystrophy (DMD) Patients

Routine clinical pharmacokinetic data collected from Duchenne muscular dystrophy (DMD) patients receiving digoxin have been analyzed to evaluate the role of patients' characteristics for estimating dosing regimens. The data were analyzed using NONMEM, a computer program designed for population pharmacokinetic analysis that allows pooling of data. The pharmacokinetic model of digoxin was described using a one-compartment steady-state model. The effect of factors on digoxin clearance was investigated. NONMEM estimates indicate that digoxin clearance was influenced by the variables of body weight and combination with diuretics. The interindividual variability in digoxin clearance was modeled with proportional error with an estimated coefficient of variation of 25%, and the intraindividual variability in digoxin concentration was modeled with equal error with an estimated standard deviation of 0.0828 ng ml(minus sign1). In order to determine whether the population parameters obtained in this study were accurate, we administered digoxin according to individual dosage regimens in four DMD patients, using these values obtained by the Bayesian method. As a result, we found that mean prediction error, which indicates the deviation of prediction accuracy for digoxin concentration in plasma, was small, as were mean absolute prediction error and root mean squared error, showing the accuracy of this prediction method. The dosing method based on clearance values obtained by NONMEM analysis allowed the prediction of the steady-state concentration as a function of maintenance dose with acceptable error for therapeutic drug monitoring.