Effects of introduced aspartic and glutamic acid residues on the P'1 substrate specificity, pH dependence and stability of carboxypeptidase Y

Carboxypeptidase Y is a serine carboxypeptidase isolated from Saccharomyces cerevisiae with a preference for C-terminal hydrophobic amino acid residues. In order to alter the inherent substrate specificity of CPD-Y into one for basic amino acid residues in P'1, we have introduced Asp and/or Glu residues at a number of selected positions within the S'1 binding site. The effects of these substitutions on the substrate specificity, pH dependence and protein stability have been evaluated. The results presented here demonstrate that it is possible to obtain significant changes in the substrate preference by introducing charged amino acids into the framework provided by an enzyme with a quite different specificity. The introduced acidic amino acid residues provide a marked pH dependence of the (kcat/Km)FA-A-R-OH/(kcat/Km)FA-A-L-OH ratio. The change in stability upon introduction of Asp/Glu residues can be correlated to the difference in the mean buried surface area between the substituted and the substituting amino acid. Thus, the effects of acidic amino acid residues on the protein stability depend upon whether the introduced amino acid protrudes from the solvent accessible surface as defined by the surrounding residues in the wild type enzyme or is submerged below.     

Identification of clinical yeast isolates by using the Microring YT

We investigated predisposing factors for the development of heterotopic ossification in a retrospective study of 637 hip arthroplasties, of which 484 were unilateral, 62 bilateral and 29 revision operations. The frequency of heterotopic ossification after a primary hip arthroplasty was 57 percent. In a univariate analysis, men, patients with hypertrophic arthrosis, and cemented arthroplasty were all at risk of developing heterotopic ossification. After a multivariate analysis, the male sex and the cemented arthroplasty remained as significant factors. In bilateral operations, the contralateral side developed heterotopic ossification in 82 percent when the primary hip operation had already caused ossification. There was no increase in ossifications after the contralateral operation. Half of the revision operations had an increase of heterotopic ossification from 1 to 4 Brooker classes.     

Effect of molecular architecture on microstructural characteristics in some polysiloxaneimide multiblock copolymers

Efforts to correlate molecular characteristics with microstructural dimensions in microphase-separated diblock and triblock copolymers have been very successful, resulting in relationships that can be utilized to design materials with a specific microstructure and, consequently, with particular thermomechanical properties. However, similar efforts in the arena of multiblock copolymers have not been nearly as extensive, despite the increasing interest and diversity of this class of materials. In the present work, energy-filtered electron microscopy (EFEM) and small-angle neutron scattering (SANS) are used in a complementary fashion to probe the phase behavior of a series of three polysiloxaneimide (PSI) multiblock copolymers with different molecular architectures. Despite their relatively short segment lengths, all three materials exhibit signs of microphase separation at ambient temperature. SANS data are obtained from chemically unaltered materials and are subsequently interpreted with the Teubner-Strey model for microemulsions. Resultant microstructural dimensions are in good agreement with those measured from EFEM micrographs. Additional insight into the intramolecular sequencing of each copolymer is obtained from scaling relationships.     

Treatment of chronic cardiac insufficiency with coenzyme Q10, results of meta-analysis in controlled clinical trials

Meta-analysis applied to eight controlled clinical trials of coenzyme Q10 (CoQ10)-treatment of congestive heart failure revealed a significant improvement a several important cardiac parameters such as ejection fraction (EF), stroke volume (SV), cardiac output (CO), cardiac index (CI) and end diastolic volume index (EDVI). Concerning the improvement in SV and CO the average patient in the CoQ10 group had a higher score than respectively 76% and 73% of the patients in the placebo group. The improvement in CO and SV was also significant when considering of homogeneity. Additional controlled clinical trials seem justified which may strengthen the power of the meta-analyses. However, based on available results, it can not be excluded that CoQ10 may have a future role a adjunctive therapy in a dosage of 100-200 mg/day in the treatment of chronic congestive heart failure.     

Epididymo-orchitis and Reiter''s disease. Two infrequent complications after intravesical bacillus Calmette-Guérin therapy

Two cases of adverse reactions to intravesical treatment with bacillus Calmette-Guérin are reported. The first patient presented with a severe suppurating epididymo-orchitis three months after his last treatment. The second patient developed Reiter's disease with arthritis of one knee and conjunctivitis 1 week after the fourth instillation.     

Contrast-medium-induced ventricular fibrillation: arrhythmogenic mechanisms and the role of antiarrhythmic drugs in dogs

RATIONALE AND OBJECTIVES: Small electrolyte additions to a nonionic contrast medium reduce the risk of ventricular fibrillation (VF) during wedged catheter injection of a contrast medium. The current study was designed to further investigate contrast-medium-induced VF by studying the effect of pretreatment with different antiarrhythmic drugs. METHODS: During a simulated wedged catheter situation, iohexol was injected into the anterior descending branch of the left coronary artery in five open-chest, anesthetized dogs pretreated with lidocaine, propranolol, amiodarone, almokalant, or verapamil. RESULTS: Wedging the catheter for 60 sec did not induce VF. However, all 15 wedged catheter injections with iohexol induced VF within 28 sec (19 +/- 1 [mean +/- standard error of the mean]) despite pretreatment with antiarrhythmic drugs. Prior to VF, conduction was slowed and monophasic action potential duration lengthened in the contrast-medium-perfused myocardium, although no significant changes occurred in the control area. CONCLUSION: The combination of catheter wedging and long-lasting contrast medium injection has a high risk of causing VF. Although adding a small amount of electrolytes to nonionic contrast media can reduce the risk of VF, antiarrhythmic drug therapy may not have a protective effect.     

Pressure-infiltration processing of reinforced aluminum

Pressure infiltration of liquid metal is one of the most important processing routes for the production of aluminum-matrix composites having a self-supporting reinforcement phase. This article briefly examines the physical phenomena governing infiltration processes, to present practical guidelines derived from their analysis for optimization of the process and the materials produced. Engineering aspects that are pertinent to infiltration techniques, including preform preparation, process configurations, flow control, and innovative processes, are summarized.     

Real-Time Global Illumination for VR Applications

Real-time global illumination in VR systems enhances scene realism by incorporating soft shadows, reflections of objects in the scene, and color bleeding. The Virtual Light Field (VLF) method enables real-time global illumination rendering in VR. The VLF has been integrated with the Extreme VR system for real-time GPU-based rendering in a Cave Automatic Virtual Environment.     

Evaluation of a type 1 diabetes serum cohort by SELDI-TOF MS protein profiling

Proteomics analysis of serum from patients with type 1 diabetes (T1D) may lead to novel biomarkers for prediction of disease and for patient monitoring. However, the serum proteome is highly sensitive to sample processing and before proteomics biomarker research serum cohorts should preferably be examined for potential bias between sample groups. SELDI‐TOF MS protein profiling was used for preliminary evaluation of a biological‐bank with 766 serum samples from 270 patients with T1D, collected at 18 different paediatric centers representing 15 countries in Europe and Japan over 2 years (2000–2002). Samples collected 1 (n = 270), 6 (n = 248), and 12 (n = 248) months after T1D diagnosis were grouped across centers and compared. The serum protein profiles varied with collection site and day of analysis; however, markers of sample processing were not systematically different between samples collected at different times after diagnosis. Three members of the apolipoprotein family increased with time in patient serum collected 1, 6, and 12 months after diagnosis (ANOVA, p<0.001). These results support the use of this serum cohort for further proteomic studies and illustrate the potential of high‐throughput MALDI/SELDI‐TOF MS protein profiling for evaluation of serum cohorts before proteomics biomarker research.