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.     

Making polyurethane foams from microemulsions

A remarkable correlation exists between the degree of expansion of polyurethane foams and the structure of the reacting premixes. Polyurethane foams obtained from reacting premixes containing microemulsions are highly expanded. The expansion rate is proportional to the volume fraction of microemulsion in the premix. The stability of premixes with and without microemulsion is completely different suggesting distinct creaming mechanisms. We apply this idea to synthesize polyurethane foams from microemulsions successfully. This approach can be used to rationalize the design of polyurethane formulations leading to highly expanded foams.     

Comparative study of the treatment of prostatic adenoma: laser photocoagulation versus endoscopic resection

OBJECTIVE: Resection of the prostate using a Nd YAG laser is designed to destroy benign prostatic hyperplasia responsible for bladder neck obstruction. This technique is currently under investigation. PATIENTS AND TECHNIQUE: A total of 56 patients, with a minimal follow-up of 6 months and presenting indications for endoscopic resection of the prostate were treated by TULIP [22] or VLAP [34] laser coagulation. The objective and subjective results were compared to a group of patients [30] treated by endoscopic resection during the same period. Results are expressed as the percentage of patient responding to treatment in terms of objective (peak flow rate > 15 ml/s with improvement > 30% or between 12 and 15 ml, but improvement > 50%) and subjective parameters (IPSS < 7 and improvement > 30% or between 7 and 10, but improvement < 50%). RESULTS: With a minimal follow-up of 6 months, the objective response rates were 55.5% (TULIP), 84.6% (VLAP) and 83.2% (TURP), while the subjective response rates were 55.5% (TULIP), 92.3% (VLAP) and 83.3% (TURP). The hospital stay was significantly shorter in the laser group (2.1 VLAP and 3.3 TULIP; 4.93 TURP). In the TULIP group, 3 patients were incontinent and 4 developed chronic prostatitis. At 6 months, 4 patients in the VLAP group and 6 patients in the TULIP group had to undergo a complementary TURP. CONCLUSION: These results indicate that laser photocoagulation is effective in the treatment of benign prostatic hyperplasia. The VLAP technique appears to be better adapted to this indication. The benefit of VLAP will be more clearly demonstrated by the randomized study currently underway, provided the length of hospital stay, postoperative complications and cost-effectiveness ratio are taken into account. In contrast, the TULIP system is associated with numerous complications which appear to be unacceptable.     

Identification and analysis of genes from Streptomyces pristinaespiralis encoding enzymes involved in the biosynthesis of the 4-dimethylamino-L-phenylalanine precursor of pristinamycin I

Dextran has been used as a carrier molecule for the synthesis of monofunctional peptide-dextran conjugates. The immunodetection of such carrier immobilized peptides on ELISA plates was compared to that of peptides adsorbed directly to immunoplates. The main features observed with peptide-dextran conjugates were as follows: only small amounts of peptide (1-2 mg) were necessary for coupling via alpha- or epsilon-amino groups to NaIO4-activated dextran (4 mg); the coupling yield was up to 68%; an amino acid analysis of the conjugate enabled the amount of carrier immobilized peptide to be calculated; an estimated 15-17 peptides were bound per dextran molecule (MW 73,500); using a carbohydrate as carrier reduces the possibility of non-specific interactions because no hydrophobic or ionic sites and no protein-like epitopes exist on the carrier apart from the peptide ligand. It can be assumed that some peptide ligands provide the forces for an interaction with the plate surface whereas other remain free for the interaction with the antibody. Thus, the detection with monoclonal anti-peptide antibodies allowed peptide-dextran conjugates to be used at coating concentrations of 1-3 nM peptide, corresponding to 0.6-2.6 ng peptide-dextran per well. In contrast, concentrations of 150-500 nM were required for coating with peptides. The applicability of monofunctional peptide-dextran conjugates was demonstrated by investigating the titer and specificity of a polyclonal anti-peptide serum developed against human gastrointestinal glutathione peroxidase. The introduction of biotin as a second ligand of the dextran conjugate permitted its capture on streptavidin coated plates. This synthesis of bifunctional peptide-biotin-dextran conjugates opens up additional possibilities for applications.     

Jitter in ATM networks and its impact on peak rate enforcement

Cells arriving to an ATM network experience random delays due to queueing in upstream multiplexing stages, notably in customer premises. This is the phenomenon of jitter and the aim of the present paper is to study its influence on peak rate enforcement. We first introduce some general characterizations of jitter and then, describe two models of jittered flows based on simple queueing systems. We discuss the objectives of peak rate enforcement and study the impact of jitter on the dimensioning of jumping window and leaky bucket mechanisms. A useful synthetic characterization of jitter appears to be a remote quantile of the cell delay distribution expressed in units of the initial inter-cell interval.     

Shear mode coupling and tilted grain growth of A1N thin films in BAW resonators.

Polycrystalline A1N thin films were deposited by RF reactive magnetron sputtering on Pt(111)/Ti electrode films. The substrates were tilted by an angle ranging from 40 degrees to 70 degrees with respect to the target normal. A low deposition temperature and a high sputter gas pressure were found ideal for tilted growth. The resulting grain tilt angle amounts to about half the substrate tilt angle. For coupling evaluation, 5 GHz solidly mounted resonator structures have been realized. The tilted grain A1N films exhibited a permittivity in the 9.5-10.5 range and loss tangent of 0.3%. Two shear modes as well as the longitudinal mode could be clearly identified. The coupling coefficient k2(eff) of the fundamental thickness shear mode (TS0) was found to be about 0.5%, which is compatible with a c-axis tilt of about 6 degrees.     

Fabrication of a biocompatible flexible electroosmosis micropump

In in-vivo microsystems, one of the components is a biocompatible micropump in order to produce the necessary force to deliver the fluid from the inlet to the outlet. In this contribution, a flexible micropump is fabricated which is aimed to be suitable in drug delivery applications. It provides high degree of biocompatibility, since the only employed materials are implantation grade polydimethylsiloxane elastomer and gold for the electrical interconnects. The working principle of the micropump is based on transverse DC electroosmosis which is a new variant of conventionally applied high voltage DC electroosmosis. This new technique is based on topography irregularities introduced in the channel resulting in a non-uniform charge distribution. The advantage is to drive the micropump using a relatively low DC voltage of 10 V while getting an effective flow speed of 60 μm/s. In order to characterize the flow speed, dyed 3 μm beads are dispersed in the working fluid and their speed is measured by the line scanning technique using a confocal microscope. It is also observed that the flow has a helical profile which is an attractive feature for an efficient micro-mixer in active microfluidics and μ-TAS applications.