Two cell-wall-associated aminopeptidases from Lactobacillus helveticus and the purification and characterization of APII from strain ITGL1

Lactobacillus helveticus ITGL1 is able to hydrolyse many amino-acyl and dipeptidyl-p-nitroanilides. Analysis of heat inactivation kinetics, metal ion and protease inhibitor effects, and the subcellular location of aminopeptidase activities in both the parental strain and mutants deficient in lysyl-p-nitroanilide hydrolysis, led to the characterization of two cell-wall-associated aminopeptidases, APII and APIV. APII, which catalysed L-lysine p-nitroanilide hydrolysis, was purified about 28-fold to homogeneity from cell-wall extracts of L. helveticus ITGL1 and characterized. The purified enzyme appeared to be monomeric, with a molecular mass of 97 kDa. Aminopeptidase activity was greatest at pH 6.5 and 50 degrees C. APII was completely inhibited by bestatin, chelating agents such as EDTA or 1,10-phenanthroline and the divalent cations Zn2+ and Cu2+. The activity of the EDTA-treated enzyme was restored by Co2+, Ca2+ or Mn2+. Although APII was able to degrade several dipeptides and tripeptides with hydrophobic N-terminal amino acid (Leu, Ala), it was inactive on peptides containing Pro or Gly, and may thus contribute to the development of cheese flavour by processing bitter peptides.     

Applications of a super element model for non-linear analysis of stiffened box structures

Numerical investigations are carried out to verify the new super element model developed in Reference 1 for non-linear analysis of stiffened box structures. The results presented here are obtained from a variety of examples which are chosen to illustrate the accuracy and convergence of the model in various applications. Good agreement is obtained for most of the problems considered and the present formulation is found to be accurate and economical for preliminary engineering design.     

The use of patent titles for identifying the topics of invention and forecasting trends

Co-word analysis applied to patents through WPIL normalized title words appears to give a useful picture of a given field: we obtain both qualitative (themes) and quantitative information (weight of themes). It also gives information about the strategic aspects of the themes. Furthermore, in some cases, it is an indication of the future of certain themes that may help forecasting and management studies. Finally, it provides information about what could be a real technology growth process, in relation to the so-called translation model used in co-word analysis.     

Kinetic analysis of papaya proteinase omega

Papaya proteinase omega (pp omega) has been purified from dried latex both by immunoaffinity and traditional methods. Kinetic analysis revealed that (1), the pp omega-catalysed hydrolysis of N-benzoyl-L-arginine p-nitroanilide (BApNA) has a lower specificity (kcat/Km) than the same reaction catalysed by papain; (2), the pp omega-catalysed hydrolysis of a tripeptide substrate having phenylalanine at the second position (S2-site) showed a more similar specificity to that catalysed by papain; (3), the significant difference between the two enzymes is that steady state kinetics with both L-BApNA and a tripeptide enables the identification in pp omega of other ionizations affecting binding. The active sites of papain and pp omega can therefore be distinguished by pH-dependence of kcat/Km.     

Determination of the radiation sensitivity of the stromal cells in the murine long-term bone marrow culture by measuring the induction and rejoining of interphase chromosome breaks

PURPOSE: To determine the radiosensitivity of bone marrow stromal cells, the rate of interphase chromosome breakage and rejoining of stromal cells in the murine long term bone marrow culture and of human skin fibroblasts were compared. METHODS AND MATERIALS: The cells were irradiated with doses up to 6 Gy and repair times up to 6 hr were investigated. After induction of premature chromosome condensation by fusing the cells with mitotic HeLa cells, the number of interphase chromosome fragments was counted. RESULTS: The number of radiation induced breaks was found to be not significantly different for both cell types with 6.16 +/- 0.26 breaks per Gray for the fibroblasts and 5.96 +/- 0.20 breaks per Gray for the stromal cells. A significant difference was observed in the repair rate. The fibroblasts rejoined 39.6% of the breaks induced initially during the first hour after irradiation and 5.6 +/- 1.84 breaks remained unrejoined after 6 hr, while the stromal cells were able to rejoin 63.2% in 1 hr and had 2.05 +/- 0.07 breaks unrejoined after 6 hr. CONCLUSION: If the well substantiated assumption is made, that the capacity to repair DNA double strand breaks or interphase chromosome breaks is correlated with the cellular radiosensitivity, this finding indicate, that murine bone marrow stromal cells are more radioresistant than human skin fibroblasts.     

Reliability and validity of the measurement of position sense in women with osteoarthritis of the knee

OBJECTIVE: We examined the test-retest reliability and the construct validity of the measurement of knee position sense for describing the functional weightbearing performance of women with osteoarthritis (OA) of the knee. METHODS: For the purpose of this study, position sense was defined as the error occurring when subjects attempted to reproduce a criterion angle in standing with visual cues eliminated. Five such tests were recorded photographically on 3 different occasions. On each occasion the 10 subjects also completed a self-paced walking test over a 13 m indoor walkway. RESULTS: The photographic measurements were reproducible (r = 0.90) and there was no change in positioning accuracy across sessions. There was a significant (p < 0.05) inverse correlation of 0.70 between the standard deviation of the mean individual measurements of position sense (precision of the test) and those of walking speed. CONCLUSION: Our study demonstrates good measurement reliability and a comparable mean angular error with repeated tests. It also suggests the amplitude of the variability of this error is a strong determinant of an individual OA patient's functional performance in walking.     

STRONG METAL-CERAMIC JOINTS

In this paper the technology for joining metals to ceramics is reviewed. The technologies of metal powder sintering, liquid phase joining with activated brazing alloys and with ceramic frit, solid-state joining, friction welding and mechanical fixation are described. In order to put some perspective on the mechanical data given in the literature, some evaluating remarks on mechanical testing are made.