The oligomeric state, complex formation, and chaperoning activity of hsp70 and hsp80 of Neurospora crassa.

Molecular chaperones perform vital cellular functions under normal growth conditions and protect cells against stress-induced damage. The stress proteins Hsp70 and Hsp80 of Neurospora crassa were extracted from heat-shocked mycelium, purified to near homogeneity, and examined with respect to their oligomeric state, complex formation, and chaperoning properties. Their oligomeric state was assessed by dynamic light-scattering measurements, and both Hsp70 and Hsp80 were observed to form a range of soluble, high-molecular-mass protein aggregates. Direct interaction between Hsp70 and Hsp80 was studied by partial tryptic digestion and surface plasmon resonance (SPR). Hsp70 was immobilized on the sensor chip surface, and the binding of Hsp80 in solution was followed in real time. Proteolytic digestion revealed that Hsp70-Hsp80 complex formation results in conformational changes in both proteins. The data from SPR studies yielded an equilibrium dissociation constant, KD, of 8.5 x 10(-9) M. The chaperoning ability of Hsp70, Hsp80, and Hsp70-Hsp80 was monitored in vitro by the protection of citrate synthase from thermal aggregation. The binding of nucleotides modulates the oligomeric state, chaperoning function, and hetero-oligomeric complex formation of Hsp70 and Hsp80.

Effects of Glimepiride vs Glibenclamide on Ischaemic Heart Disease Risk Factors and Glycaemic Control in Patients with Type 2 Diabetes Mellitus.

OBJECTIVE: This study aimed to compare the effects of glimepiride and glibenclamide on glycaemic control and a range of risk factors for ischaemic heart disease (IHD), including concentrations of insulin-like molecules. PATIENTS: A double-blind, placebo-controlled, randomised, crossover comparison of 4 weeks of treatment with glibenclamide 2.5 to 20 mg/day and glimepiride 1 to 8 mg/day was undertaken in 29 type 2 (non-insulin-dependent) diabetic patients. The average (mean +/- SD) duration of diabetes was 8.5 (+/- 5.9) years. RESULTS: Compared with placebo, fasting plasma glucose was significantly lower on both drugs [placebo (P): mean (SD) 11.9 (3.3) mmol/L, glibenclamide: 9.5 (3.2); p < 0.0005, glimepiride: 10.6 (3.4); p = 0.01] and lower on glibenclamide than glimepiride (p = 0.003). The integrated, meal-stimulated rise in glucose was lower with glimepiride, but not glibenclamide, compared with placebo [P: 588.1 (372.2) mmol/L.min, glimepiride: 443.0 (346.9) mmol/L.min; p = 0.010, glibenclamide: 586.4 (366.2) mmol/L.min; p = 0.630]. There was no between-drug difference (p = 0.145). Fasting insulin did not differ compared with placebo [P: 92.3 (61.3) pmol/L, glimepiride: 91.8 (60.6) pmol/L; p = 0.787, glibenclamide: 87.8 (51.6) pmol/L; p = 0.379] and there was no between-drug difference (p = 0.601). There were no significant differences in effect upon fasting concentrations of C-peptide, proinsulin, des 31,32 proinsulin or the ratio of proinsulin-like to total insulin-like molecules. The integrated insulin and C-peptide responses to a meal were significantly greater on both drugs than on placebo [insulin: median (25th, 75th percentile), P: 7073 (2430-18296) pmol/L.min, glibenclamide: 18045 (4290-35850) pmol/L.min; p = 0.0005, glimepiride: 14355 (5880-32820) pmol/L.min; p = 0.0001; C-peptide mean (SD): P: 51.89 (49.01) nmol/L.min, glibenclamide: 90.15 (59.44) nmol/L.min; p = 0.006, glimepiride: 89.75 (61.78) nmol/L.min; p = 0.007], but there was no between-drug difference [integrated insulin (p = 0.923), integrated C-peptide (p = 0.680)]. Compared with placebo, plasminogen activator inhibitor (PAI) antigen was significantly lower on glibenclamide but not glimepiride [P: 28.8 (19.7) microg/L, glimepiride: 24.4 (15.2) microg/L; p = 0.300, glibenclamide: 20.0 (10.9) microg/L; p = 0.003]. PAI activity was similar with all agents, as was low density lipoprotein (LDL)-cholesterol [P: 4.4 (1.2) mmol/L, glimepiride: 4.2 (0.9) mmol/L; p = 0.225, glibenclamide: 4.5 (1.4) mmol/L; p = 0.174]. Corrected for fasting plasma glucose, LDL was 0.5 mmol/L lower on glimepiride than on glibenclamide (95% confidence interval: -0.8, -0.2), a clinically significant difference. There were no significant differences in other measured factors. CONCLUSION: Both drugs improved glycaemia without adversely affecting a range of IHD risk factors.

Improving job satisfaction and enlarging the role of technicians.

The process used in preparing this administrative report provided the means of documenting departmental support of technician task expansion, cost benefits, increased availability of professional time, and the need for a dynamic department. Some of the benefits to St. Luke's Regional Medical Center and its Pharmacy Department were an increase in technician job satisfaction, improved quality of patient care attributable to the increased scope of pharmacy service, and up to 8554 hours per year of professional time available for more clinically oriented responsibilities. If technician turnover were decreased by half, technician pricing errors eliminated, and technicians assumed the identified technical tasks, potential monetary benefits could be as much as +116,900 per year. The departmental cost of implementing these recommendations would be an additional 4.16 technician FTEs, and the professional time for training, supervising, and coordinating technicians and functions. These recommendations were well received by pharmacy administration, and the committee was asked to develop a list of priorities and an implementation plan for administrative approval.

Effects of fish oil supplements in NIDDM subjects. Controlled study.

The aim of this study was to evaluate the effects of a fish oil preparation (MaxEPA) on hemostatic function and fasting lipid and glucose levels in non-insulin-dependent diabetic (NIDDM) subjects. Eighty NIDDM outpatients aged 55.9 yr (mean SD 11.5 yr) participated in a prospective double-blind placebo-controlled study of MaxEPA capsules (10 g/day) or olive oil (control) treatment over 6 wk. Patients received either MaxEPA or olive oil in addition to preexisting therapy. Metabolic and hemostatic variables were measured before treatment and after 3 and 6 wk. Platelet membrane eicosapentaenoic acid (EPA) content increased in the treatment group (P less than 0.001). MaxEPA supplementation was associated with a significant fall in total triglycerides (P less than 0.001) but did not affect total cholesterol (P = 0.7) compared with control treatment. Fasting plasma glucose increased after 3 wk (P = 0.01) but not after 6 wk (P = 0.17) treatment with MaxEPA. Spontaneous platelet aggregation in whole blood fell in the MaxEPA group (P less than 0.02) after 6 wk, but there were no changes in agonist-induced platelet aggregation, thromboxane generation in platelet-rich plasma, or plasma beta-thromboglobulin and platelet factor IV levels. An increase in clotting factor VII (P = 0.02), without changes in fibrinogen or factor X levels, occurred in the MaxEPA group. Similar reductions in blood pressure were observed in both groups. Dietary supplementation with MaxEPA capsules (10 g/day) in NIDDM subjects is associated with improvement in hypertriglyceridemia but with deleterious effects in factor VII and blood glucose levels. Most indices of platelet function are unaffected by this therapy.