Purification and characterization of an extracellular lipase from Penicillium candidum

Penicillium candidum produces and secretes a single extracellular lipase with a monomer molecular weight of 29 kDa. However, this enzyme forms dimers and higher molecular weight aggregates under nondenaturing conditions. The lipase from P. candidum was purified 37-fold using Octyl-Sepharose CL-4B and DEAE-Sephadex columns. The optimal assay conditions for lipase activity were 35°C and pH 9. The lipase was stable in the pH range of 5–6 with a pl of 5.5, but rapid loss of the enzyme activity was observed above 25°C. Tributyrin was found to be the best substrate for the P. candidum lipase, among those tested. Metal ions such as Fe²⁺ and Cu²⁺ inhibited enzymatic activity and only Ca²⁺ was able to slightly enhance lipase activity. Ionic detergents inhibited the activity of the enzyme, whereas nonionic detergents stimulated lipase activity.     

Impaired IL-1beta-induced neutrophil accumulation in tachykinin NK1 receptor knockout mice

Tachykinin NK1 receptors play an important role in the development of neurogenic inflammatory responses. We have used the murine air-pouch model to investigate whether the neurogenic component of the cellular inflammatory response to interleukin-1beta (IL-1beta, 10 ng into the air-pouch) is altered in NK1 receptor knockout mice compared to wild type controls. Air-pouches were washed following a 4 h IL-1beta treatment, the wash collected and neutrophil number estimated using a Neubauer haemocytometer. The response to IL-1beta was significantly attenuated in NK1 receptor +/- (40% reduction) and -/- mice (62% reduction) compared to wild type controls (+/+), whilst the response to cytokine-induced neutrophil chemoattractant (CINC, 0.3 microg) was unaffected. The response to substance P (7.5 nmol) was attenuated by approximately 50% in both NK1 receptor +/- and -/- mice compared to wild type controls. In conclusion NK1 receptors play a significant role in the cellular response to IL-1beta in a model of inflammation.     

Evaluation of the Dacos 3.0 analyser

The selective multitest Coulter Dacos 3.0 analyser was evaluated according to the guidelines of the Comisión de Instrumentación de la Sociedad Española de Química Clínica and of the European Committee for Clinical Laboratory Standards.The evaluation was performed in four steps: examination of the analytical units; evaluation of routine working; study of interferences; and assessment of practicability.The evaluation included a photometric study. The inaccuracy is acceptable for 340 nm and 420 nm, and the imprecision at absorbances from 0.05 to 2.00 ranged from 0.06 to 0.28% at 340 nm and from 0.06 to 0.08% at 420 nm. The linearity showed some dispersion at low absorbance for PNP at 420 nm and the drift was negligible.The imprecision of the pipette delivery system, the temperature control system and the washing system were satisfactory.In routine work conditions, seven analytical methods were studied: glucose, creatinine, iron, total protein, AST, ALP and calcium. Within-run imprecision ranged, at low concentrations, from 0.9% (CV) for glucose, to 7.6% (CV) for iron; at medium concentrations, from 0.7% (CV) for total protein to 5.2% (CV) to creatinine; and at high concentrations, it ranged from 0.6% (CV) for glucose to 3.9% (CV) for ALP.Between-run imprecision at low concentrations ranged from 1.4% (CV) for glucose to 15.1% (CV) for iron; at medium concentrations it ranged from 1.2% (CV) for protein to 6.7% (CV) for iron; and at high concentrations the range is from l.2for AST to 5.7% (CV) for iron.No contamination was found in the sample carry-over study. Some contamination was found in the reagent carry-over study (total protein due to iron and calcium reagents). Relative inaccuracy is good for all the constituents assayed. Only LDH (high and low levels) and urate (low level) showed weak and negative interference caused by turbidity, and γ-GT (high level) and amylase, bilirubin and ALP (two levels) showed a negative interference caused by haemolysis.     

Cross-linkable polyester imides

Aromatic polyesters containing imide pendant groups have been prepared from diphenols and 5-imidoisophthaloyl chlorides by the interfacial polycondensation method. Linear polyesters without substituents have also been synthesized by the same method for comparative purposes. The presence of side imide groups enhances the solubility of aromatic polyesters in organic solvents, and the thermal properties are greatly affected, with increases in the glass transition temperatures of up to 60°C, while the thermal resistance (t.g.a.) seems not to be so greatly affected. When the imide pendant groups are unsaturated, as derived from maleimide, nadimide and tetrahydrophthalimide, the polyesters can be converted into crosslinked materials by means of the appropriate thermal treatment, either as a polymeric powder or as films. Properties such as solubility, thermal resistance and mechanical strength were investigated before and after the cross-linking reaction.     

Approach to the analysis of the dynamics of industrial FCC units

Although it is known that FCC units are operated at a controllable steady state, it is commonly assumed that this state is pseudostable. An ignited steady state, theoretically stable, outside the operable domain has also been reported for the same set of operating parameters. The aim of this work is to introduce a theoretical method for the preliminary analysis of the controllability of an industrial FCC unit, at any set of conditions, using a nonlinear model directly. An approach for the analysis of the stability of the system zero dynamics is used to investigate the dynamic behaviour of two common operating policies when the unit is operating either in the standard or in the ignited steady state. It is used to predict some particular characteristics of the process, such as the presence of inverse response after the change in catalyst circulation rate, a common control action. The results derived from the analysis developed are discussed in terms of physical responses and verified by performing open loop dynamic simulations of an industrial FCC unit. The operation of the unit is simulated for both the standard and the ignited steady states.     

Evaluation of the Hitachi 717 analyser

The selective multitest Boehringer Mannheim Hitachi 717 analyser was evaluated according to the guidelines of the Comisión de Instrumentación de la Sociedad Española de Química Clinica and the European Committee for Clinical Laboratory Standards. The evaluation was performed in two steps: examination of the analytical units and evaluation in routine operation.The evaluation of the analytical units included a photometric study: the inaccuracy is acceptable for 340 and 405 nm; the imprecision ranges from 0.12 to 0.95% at 340 nm and from 0.30 to 0.73 at 405 nm, the linearity shows some dispersion at low absorbance for NADH at 340 nm, the drift is negligible, the imprecision of the pipette delivery system increases when the sample pipette operates with 3 μl, the reagent pipette imprecision is acceptable and the temperature control system is good.Under routine working conditions, seven determinations were studied: glucose, creatinine, iron, total protein, AST, ALP and calcium. The within-run imprecision (CV) ranged from 0.6% for total protein and AST to 6.9% for iron. The between run imprecision ranged from 2.4% for glucose to 9.7% for iron. Some contamination was found in the carry-over study. The relative inaccuracy is good for all the constituents assayed.     

Functional expression of the Erwinia uredovora carotenoid biosynthesis gene crtl in transgenic plants showing an increase of beta-carotene biosynthesis activity and resistance to the bleaching herbicide norflurazon

The possibility that extracellular matrix-cell surface interaction might lead to signalling was examined in human mononuclear cells. In mononuclear cells loaded with Indo-1 acetoxymethylester, collagen binding elicited a 2-3 fold increase in cytosolic free calcium concentration within seconds. Plasma fibronectin, serum albumin and heparin did not cause any significant change in intracellular calcium levels. Collagen I, collagen IV and glucosylated collagen I evoked an increase in intracellular Ca++ both in the presence and absence of extracellular calcium. However, in the presence of 2mM EDTA, the increase in cytosolic free calcium concentration caused by collagen I and collagen IV was partly abolished, suggesting the requirement of a cation-dependent interaction of collagen with mononuclear cells. Glucosylated collagen induced intracellular calcium mobilization even in the presence of EDTA suggesting a cation-independent interaction. These results indicate that collagen binding induces rapid mobilization of calcium in human mononuclear cells, apparently from intracellular sources.     

Nutrient transfers by leaching in a no-tillage system through soil treated with repeated pig slurry applications

Repeated pig slurry applications cause accumulation and leaching of soil nutrients and, subsequently, groundwater contamination. The purpose of this study was to evaluate ammonium (NH₄ ⁺–N), nitrate (NO₃ ^?–N), phosphorus (P) and potassium (K) leaching in a sandy soil with a 5-year history of repeated pig slurry applications. The study was carried out in the experimental field of the Universidade Federal de Santa Maria (UFSM) (Federal University of Santa Maria), Santa Maria, Rio Grande do Sul, in the South of Brazil, from 2002 to 2007, in no-tillage system in a Typic Hapludalf soil. Slurry was applied at doses of 0, 20, 40 and 80 m³ ha^?1, which over the 5 years amounted to the addition of 594, 1,188 and 2,376 kg N ha^?1; 508, 1,016 and 2,032 kg P ha^?1 and 216, 432 and 864 kg K ha^?1, respectively. Leachate solutions were collected throughout the period, prepared and then subjected to analysis of NH₄ ⁺–N, NO₃ ^?–N, P and K available in the solution. Repeated applications of pig slurry in a no-tillage system in sandy soil led to the transfer of elements through the leachate solution according to the sequence: nitrate > potassium > ammonium > phosphorus. The transfers of these elements were positively related to the increased volume of leachate solution over the years in addition to the nutrient amounts added by the pig slurry applied over the 60-month period.