Biochemical and molecular biological characterization of a lipase produced by the fungus Rhizopus delemar

The results of a comprehensive biochemical and molecular biological investigation of the lipase produced by the mycelial fungus Rhizopus delemar are described. This enzyme cleaves and synthesizes primary esters and related bonds, exhibits 1,3-positional selectivity in its actions on glycerides, and is a member of a family of enzymes that have been widely used in applied biocatalysis. Use of glycerol as main carbon source rather than glucose or lipid supported mycelial growth and lipase production. The enzyme was purified to homogeneity and characterized. Pure lipase was crystallized and its three-dimensional structure determined. The enzyme was found to adopt a configuration similar to those of other members of its homologous family. The structural data also indicated that lipases possess greater configurational mobility than had been previously appreciated. A complementary DNA clone was isolated that contained the full length lipase gene. The nucleic acid sequence of this cDNA indicated that it was initially synthesized as a preproenzyme, and allowed determination of the complete predicted amino acid sequence of the lipase, and its comparison to the sequences of related enzymes. Truncated forms of the cloned cDNA were produced that encoded either mature or prepro-lipase. These DNAs were introduced into a tightly regulated E. coli expression system, overcoming the toxicity of the enzyme while also allowing overproduction of lipase. Molecular modelling was employed to guide the rational mutagenesis of the enzyme, identifying sites within the substrate binding region that regulated substrate selectivity. Mutant lipases were generated with altered substrate specificities, creating novel enzymes and beginning the definition of structure-function relationships in the lipolytic enzymes.     

Hydromorphological consequences of regulating the lower vistula,Poland

Quantitative and qualitative data on channel changes along the lower Vistula arising from regulation works carried out in the 19th and 20th centuries are presented. Channel width was reduced by 50% and the bed lowered by 1.3 m. Changes of the river's hydrodynamic conditions in the segment from Wloclawek to Swiecie during the last 140 years are discussed. Changes of the bed load balance are characterized.     

SIMULATION OF RF-ASSISTED CONVECTCVE DRYING FOR SEED QUALITY BROAD BEAN

Based on experimental data and analysis of the heat and mass transfer mechanism, a semi-empirical model for dielectrically-assisted convective drying of shrinkable and hygroscopic materials with internal resistance to mass flow has been derived and used to simulate radio-frequency drying of the seed quality broad bean (Vicia Faba)¹.     

Limited-lifetime shared-access in mobile systems

In this paper, we propose a simple access protocol to shared information in a mobile environment. The objective of the proposed scheme is to allow a specified set of users access toshared information and protect the confidentiality of the information from users outside this set. The set of users may be updated from time to time. In particular, the duration during which a user is allowed access to the shared information may be restricted. Furthermore, the information itself has limited lifetime and the confidentiality of the information has to be preserved during this lifetime only. The proposed access protocol is in particular suited to the mobile environment, because of the loose binding of the communicating entities.     

TTL Prediction Schemes and the Effects of Inter-Update Time Distribution on Wireless Data Access

Modern mobile networks, such as GPRS and UMTS, support wireless data applications. One successful example is the ever popular i-Mode in Japan. Wireless data services (wireless Internet) become more important as more and more customers of handheld devices enjoy the convenience of the ubiquitous computing. To improve the effective wireless data access, the time-to-live (TTL) management for data entries becomes important due to its use in effective caching design. In this paper, we study three TTL prediction schemes and investigate the effects of the inter-update time distribution on the wireless data access. Performance analysis is carried out via simulations as well as analytical modeling. We expect our results will be useful for the future wireless data access systems, in which transmission power for mobile devices is more limited.     

Characterization of the potassium channels involved in EDHF-mediated relaxation in cerebral arteries

1. In the presence of NG-nitro-L-arginine (L-NOARG, 0.3 mM) and indomethacin (10 microM), the relaxations induced by acetylcholine and the calcium (Ca) ionophore A23187 are considered to be mediated by endothelium-derived hyperpolarizing factor (EDHF) in the guinea-pig basilar artery. 2. Inhibitors of adenosine 5'-triphosphate (ATP)-sensitive potassium (K)-channels (KATP; glibenclamide, 10 microM), voltage-sensitive K-channels (Kv; dendrotoxin-1, 0.1 microM or 4-aminopyridine, 1 mM), small (SKCa; apamin, 0.1 microM) and large (BKCa; iberiotoxin, 0.1 microM) conductance Ca-sensitive K-channels did not affect the L-NOARG/indomethacin-resistant relaxation induced by acetylcholine. 3. Synthetic charybdotoxin (0.1 microM), an inhibitor of BKCa and Kv, caused a rightward shift of the concentration-response curve for acetylcholine and reduced the maximal relaxation in the presence of L-NOARG and indomethacin, whereas the relaxation induced by A23187 was not significantly inhibited. 4. A combination of charybdotoxin (0.1 microM) and apamin (0.1 microM) abolished the L-NOARG/ indomethacin-resistant relaxations induced by acetylcholine and A23187. However, the acetylcholine-induced relaxation was not affected by a combination of iberiotoxin (0.1 microM) and apamin (0.1 microM). 5. Ciclazindol (10 microM), an inhibitor of Kv in rat portal vein smooth muscle, inhibited the L-NOARG/ indomethacin-resistant relaxations induced by acetylcholine and A23187, and the relaxations were abolished when ciclazindol (10 microM) was combined with apamin (0.1 microM). 6. Human pial arteries from two out of four patients displayed an L-NOARG/indomethacin-resistant relaxation in response to substance P. This relaxation was abolished in both cases by pretreatment with the combination of charybdotoxin (0.1 microM) and apamin (0.1 microM), whereas each toxin had little effect alone. 7. The results suggest that Kv, but not KATP and BKCa, is involved in the EDHF-mediated relaxation in the guinea-pig basilar artery. The synergistic action of apamin and charybdotoxin (or ciclazindol) could indicate that both Kv and SKCa are activated by EDHF. However, a single type of K-channel, which may be structurally related to Kv and allosterically regulated by apamin, could also be the target for EDHF.     

Study of Polyurethane Foaming Dynamics Using a Heat Flow Meter

This work presents the results of the study concerning the effects of fillers addition on the heat flux density \(\dot{q}\left( t \right) \) of foaming of polyurethane–polystyrene porous composite (PSUR) and describes the dynamics of this process during the first 600 s. This foaming process resulted in obtaining porous materials that were based on HFC 365/225 blown rigid polyurethane foam (PUR) matrix, which contained thermoplastic expandable polystyrene (EPS) beads as the filler. In PSUR composites, the EPS beads were expanded after being heated to a temperature above the glass transition temperature of EPS and vaporing gas incorporated inside, by using the heat of exothermic reaction of polyol with isocyanate. From the start \((t=0)\) to the end of the PSUR composite foaming process (\(t=t_{k}), \dot{q}\left( t \right) \) was measured with the use of the heat flow meter. For the purpose of the study two PUR systems were selected: one with high and one with low heat density of foaming process q. EPS beads were selected from the same manufacturer with large and small diameter. The mass fraction of EPS in PSUR foam varied during the measurements. Additionally, a study of volume fractions of expanded EPS phase in PSUR foams as a function of mass fractions of EPS was conducted. In order to verify effects of the EPS addition on the heat flux density during PSUR foaming process, the thermal conductivity measurements were taken.