Purification and characterization of a novel ceramidase from Pseudomonas aeruginosa

We report here a novel type of ceramidase of Pseudomonas aeruginosa AN17 isolated from the skin of a patient with atopic dermatitis. The enzyme was purified 83,400-fold with an overall yield of 21.1% from a culture supernatant of strain AN17. After being stained with a silver staining solution, the purified enzyme showed a single protein band, and its molecular mass was estimated to be 70 kDa on SDS-polyacrylamide gel electrophoresis. The enzyme showed quite wide specificity for various ceramides, i.e. it hydrolyzed ceramides containing C12:0-C18:0 fatty acids and 7-nitrobenz-2-oxa-1, 3-diazole-labeled dodecanoic acid, and not only ceramide containing sphingosine (d18:1) or sphinganine (d18:0) but also phytosphingosine (t18:0) as the long-chain base. However, the enzyme did not hydrolyze galactosylceramide, sulfatide, GM1, or sphingomyelin, and thus was clearly distinguished from a Pseudomonas sphingolipid ceramide N-deacylase (Ito, M., Kurita, T., and Kita, K. (1995) J. Biol. Chem. 270, 24370-24374). This bacterial ceramidase had a pH optimum of 8.0-9.0, an apparent Km of 139 microM, and a Vmax of 5.3 micromol/min/mg using N-palmitoylsphingosine as the substrate. The enzyme appears to require Ca2+ for expression of the activity. Interestingly, the 70-kDa protein catalyzed a reversible reaction in which the N-acyl linkage of ceramide was either cleaved or synthesized. Our study demonstrated that ceramidase is widely distributed from bacteria to mammals.     

Production of fungal lipases using wheat bran and soybean bran and incorporation of sugarcane bagasse as a co-substrate in solid-state fermentation

Fungal strains were screened for lipase producing activities and 10 strains were classified as good producers. Aspergillus sp., Fusarium sp., and Penicillium sp. exhibited the highest activities when fermented in wheat bran (WB) and soybean bran (SB). No fungal growth was observed using sugarcane bagasse (CB). An experimental design was applied to incorporate CB into the fermentation process for lipase production by Aspergillus sp. and Penicillium sp., and to evaluate the best moisture content for the substrate. Strains studied achieved maximum lipase activities with 25% CB combined with 75% WB or SB at 40% moisture content. The highest lipase activities were observed for WB and SB, and for SB combined with CB using Aspergillus sp. Fermentation of 96 h was the optimum period for enzyme production.     

Properties of oriented strandboard made of wood species from Brazilian planted forests: Part 1: 80 mm-long strands of Pinus taeda L.

This research is part of a general study on the properties of oriented strandboard (OSB) using wood strands of species from Brazilian planted forests. The OSB industry is the latest wood related activity established in Brazil. In this particular part of the study, 80 mm long strands of Pinus taeda L. were bonded using two resin types (urea-formaldehyde and phenol-formaldehyde) at two levels of resin content (5% and 8%) to produce three-layer cross-aligned OSB to a face to core layer ratio of 1:2 and target density of 0.75 g/cm³. Physical and mechanical properties of the boards were evaluated according to ASTM standard D 1037-96a (1997) and the results compared to standards available as requirements for commercial structural panels. The results indicate that all the mechanical properties evaluated were above the requirements set forth by the Canadian standard CSA O437.0 (1993) for structural panels. The results of Janka hardness were in average 4 folds higher than the minimal requirements for Grade R-1 waferboard. Screw withdrawal values were also above the minimum required by grade M-3 of ANSI A208.1 standard (1993). Nevertheless, values of thickness swelling and water absorption were very high. The low dimensional stability may be related to the high density of the boards (“springback” effect) and also to the fact that no wax was used.     

Probabilistic models considering dependent relation in reasoning for decision-making

This paper proposes a fuzzy dependence-index for construction of the probabilistic models considering dependent relation for solving the reasoning problem. It is important for constructing the joint probability-distribution to consider the dependency of events. We consider that some vagueness is included in the dependency. Because causal relationship of among events is uncertain, it is difficult to express dependency as definite value. In this paper, we classify the dependent relations, and apply the fuzzy probability to calculation of the dependence-index. Then, the fuzzy dependence-index is defined to consider dependency with fuzziness. Using the fuzzy dependence-index, we calculate the joint probability of multi-events for constructing the probabilistic model. This work was presented in part at the 13th International Symposium on Artificial Life and Robotics, Oita, Japan, January 31–February 2, 2008     

cDNA cloning, tissue distribution, and subcellular localization of horseshoe crab big defensin

A full-length cDNA for horseshoe crab big defensin with a strong antimicrobial activity was obtained from a hemocyte cDNA library. The open reading frame of the cDNA coded for an NH2-terminal signal sequence followed by a propeptide and the mature big defensin. The propeptide is linked to the mature protein through an -Arg-X-Lys/Arg-Arg- motif, the processing site for Kex2-like proteases. Northern blot analysis revealed that big defensin is expressed in all the tissues tested, suggesting that big defensin plays an important role not only in hemocytes but also in other tissues for host defense. The subcellular localization, determined by immunocytochemistry at ultrastructural level, confirmed the previous findings obtained by biochemical analysis that big defensin locates in both small and large granules in hemocytes. Big defensin is the first example to demonstrate the existence of broad tissue distribution in horseshoe crab.     

Optimal service control of a station connected with two parallelsubstations

The optimization problem is studied for a queueing network consisting of one station and two parallel subordinate stations. After service completion at the first station, the job is divided into two subjobs and each of them enters one of the subordinate stations. The service rate at the first station is controlled in order to minimize the expected total discounted cost or the long-run average expected cost     

Tensile stress in journal bearings

The precise pressure distribution in the range of negative pressure of journal bearings was measured dynamically by a semiconductor pressure transducer fitted in the journal. Characteristics of the pressure distribution in the range of negative pressure were investigated and tensile stress was detected. A special apparatus was constructed to investigate the characteristics of tensile stress generation in lubricating oils. Currently available mineral oils of low viscosity, within a short standing time, were in a condition under which tensile stress could be generated.     

Direct synthesis of L10 type Fe-Pt nanoparticles using microwave-polyol method

We report the synthesis of Fe-Pt nanoparticles with microwave irradiation during polyol-reduction reaction. Chemically ordered Fe-Pt nanoparticles with L1₀ structure are fabricated at 250 °C using a microwave-polyol method without any post-synthesis treatments. Mössbauer analyses reveal the nanoparticles have partially ordered L1₀ structure. The partially ordered Fe-Pt nanoparticles exhibit coercivity of 3.4 kOe, saturation magnetization of 49 emu/g, and anisotropy field of 83 kOe at room temperature.     

Four-point probe resistance measurements using PtIr-coated carbon nanotube tips

We performed four-terminal conductivity measurements on a CoSi2 nanowire (NW) at room temperature by using PtIr-coated carbon nanotube (CNT) tips in a four-tip scanning tunneling microscope. The physical stability and high aspect ratio of the CNT tips made it possible to reduce the probe spacing down to ca. 30 nm. The probe-spacing dependence of resistance showed diffusive transport even at 30 nm and no current leakage to the Si substrate.