Isolation and characterization of three Dictyostelium myosin-I isozymes

Using ion exchange chromatography and an ATP-dependent actin precipitation step, we have isolated three myosin-I isozymes that, together, account for most of the K+EDTA-ATPase activity recovered from extracts of Dictyostelium. The two major myosin-I isozymes, present in approximately equal amounts, had apparent molecular masses of 125 kDa on SDS gels and have been identified by amino acid sequence analysis as the products of the Dictyostelium myosin-IB (DMIB) and myosin-ID (DMID) genes. DMIB, with a specific K+EDTA-ATPase activity 10-fold higher than DMID, was responsible for most of the activity in cell extracts. The third isozyme, present in low amounts, had an apparent molecular mass of 137 kDa on SDS gels and is too large to be the product of any of the known myosin-I genes. DMIB eluted from DE53 cellulose columns as two distinct peaks (II and III). Addition of the phosphatase inhibitor okadaic acid to the extraction buffer increased the fraction of DMIB recovered from growth phase cells in peak III from 35 to 70%. DMIB isolated from peak III, but not from peak II, displayed a significant level of actin-activated MgATPase activity. These results indicate that peak III represents a phosphorylated, actin-activatable form of DMIB.     

Survival of Yersinia enterocolitica and Escherichia coli in spring water

Survival of pure and mixed cultures of Yersinia enterocolitica and Escherichia coli in sterile spring water stored at 4 degrees C was evaluated. Y. enterocolitica increased two or three logs during the first 3 weeks of incubation, and at the end of 64 weeks viable cells in numbers corresponding to the initial inoculum level were still detectable. However, after 1 week of incubation, the number of E. coli in water started to decrease and in 13 weeks time no E. coli was detected. More rapid decrease of E. coli was observed in mixed cultures.     

Application of Romero García method to the degradation process of DGEBA/ETDA filled with ultra fine metallic copper

Summary The thermal properties of an epoxy resin (diglycidyl ether Bisphenol A, DGEBA) cured with a primary amine (ethylenediamine, ETDA) and filled with 10, 20 and 30 wt. % of ultra fine copper particles were analyzed. The thermal results were evaluated by means of the Romero-Garcia method, which allowed to obtain the resin degradation kinetic parameters, as well as the possible decomposition mechanism. From the obtained results is possible to infer that higger copper content strongly affects the degradation process of the epoxy resin, which causes a drop off on the composites thermal stability. This behavior could be attributed to water presence, being this effect less pronounced for the resin with 10 wt. % copper and unfilled resin. Regarding to decomposition mechanism, three way transport showed the best correlation for all samples.     

Ecophysiology of ochratoxigenic Aspergillus ochraceus and Penicillium verrucosum isolates. Predictive models for fungal spoilage prevention - a review

Ochratoxin A (OTA) is a secondary metabolite produced by several species of Aspergillus and Penicillium; among them Aspergillus ochraceus and Penicillium verrucosum are two ochratoxigenic species capable of growing in different climates and thus contamination of food crops with OTA can occur worldwide. OTA can be found in a wide range of foods such as cereals, coffee, cocoa, spices, beer, wine, dried vine fruit, grapes and meat products. OTA is toxic to animals, it presents neurotoxic, immunotoxic and nephrotoxic effects. It has been implicated in a human kidney disorder known as Balkan Endemic Nephropathy. This review focuses on the ecophysiology of ochratoxin-producing Aspergillus ochraceus and Penicillium verrucosum, the effect of environmental factors on their germination, mycelial growth, and OTA production. Knowledge of environmental conditions required for sucessive stages of fungal development represent the first step towards preventing mycotoxin formation. Predictive models for different stages of fungal development are presented, which allow prediction of the time before spoilage as a function of the abiotic factors. Finally, the implications of these studies in management of barley, coffee and grapes are described. This can help to identify the critical control points in their production, storage and distribution processes.