Simulation methods with extended stability for stiff biochemical Kinetics

Background  

With increasing computer power, simulating the dynamics of complex systems in chemistry and biology is becoming increasingly routine. The modelling of individual reactions in (bio)chemical systems involves a large number of random events that can be simulated by the stochastic simulation algorithm (SSA). The key quantity is the step size, or waiting time, τ, whose value inversely depends on the size of the propensities of the different channel reactions and which needs to be re-evaluated after every firing event. Such a discrete event simulation may be extremely expensive, in particular for stiff systems where τ can be very short due to the fast kinetics of some of the channel reactions. Several alternative methods have been put forward to increase the integration step size. The so-called τ-leap approach takes a larger step size by allowing all the reactions to fire, from a Poisson or Binomial distribution, within that step. Although the expected value for the different species in the reactive system is maintained with respect to more precise methods, the variance at steady state can suffer from large errors as τ grows.     

The cell cycle modulated glycoprotein GP115 is one of the major yeast proteins containing glycosylphosphatidylinositol

The cell cycle modulated protein gp115 (115 kDa, isoelectric point about 4.8-5) of Saccharomyces cerevisiae undergoes various post-translational modifications. It is N-glycosylated during its maturation along the secretory pathway where an intermediary precursor of 100 kDa (p100), dynamically related to the mature gp115 protein, is detected at the level of endoplasmic reticulum. Moreover, we have shown by the use of metabolic labeling with [35S]methionine, [3H]palmitic acid and myo-[3H]inositol combined with high resolution two-dimensional gel electrophoresis and immunoprecipitation with a specific antiserum, that gp115 is one of the major palmitate- and inositol-containing proteins in yeast. These results, and the susceptibility of gp115 to phosphatidylinositol-specific phospholipase C treatment strongly indicate that gp115 contains the glycosylphosphatidylinositol (GPI) structure as membrane anchor domain. The two-dimensional analysis of the palmitate- and inositol-labeled proteins has also allowed the characterization of other polypeptides which possibly contain a GPI structure.     

The role of humans in the diversification of a threatened island raptor

Background  

Anthropogenic habitat modifications have led to the extinction of many species and have favoured the expansion of others. Nonetheless, the possible role of humans as a diversifying force in vertebrate evolution has rarely been considered, especially for species with long generation times. We examine the influence that humans have had on the colonization and phenotypic and genetic differentiation of an insular population of a long-lived raptor species, the Egyptian vulture (Neophron percnopterus).     

Temperature cycles induce a bimodal activity pattern in ruin lizards: masking or clock-controlled event? A seasonal problem.

The daily locomotor activity pattern of Ruin lizards in the field is mainly unimodal, except for summer months when soil temperatures exceed 40 degrees C to 42 degrees C around midday. In such a situation, lizards reduce their locomotor activity around midday to avoid overheating, and thus their daily activity pattern becomes bimodal. The bimodal pattern expressed in the field is usually retained in the free-running rhythm under constant temperature and DD for a couple of weeks, after which the bimodal pattern changes into a unimodal pattern. In the present study, the authors examined whether 24-h temperature cycles (TCs) would change lizard activity from a unimodal to a bimodal pattern. Administration of TCs to unimodal lizards free-running in DD is able to entrain locomotor rhythms and to induce a bimodal pattern both in summer and autumn-winter. There are, however, striking seasonal differences in the effectiveness with which TCs achieve bimodality: (a) Numbers of lizards rendered bimodal are significantly higher in summer than in autumn-winter; (b) TCs require less time to achieve bimodality in summer than autumn-winter; (c) bimodality is retained as an aftereffect in the postentrainment free-run in summer, but not in autumn-winter; (d) TCs change activity duration in summer, but not in autumn-winter. All this demonstrates the existence of seasonal changes in responsiveness of the circadian oscillators controlling activity to the external factors inducing bimodality. Oscillators' responsiveness is high in summer, when bimodality is the survival strategy of Ruin lizards to avoid overheating around midday in open fields, and low in autumn-winter, when bimodality has no recognizable adaptive significance.     

A rapid extraction method for detecting aflatoxin producing isolates

A rapid extraction method for screening aflatoxin producing potential ofAspergillus flavus group isolates is described. The method is performed using a moist wheat medium with ca. five infected grains extracted with 2 mL of chloroform, and using thin layer chromatography. This method was proved with 95A. flavus isolates from animal feeds.     

Isolation of apical plasma membrane in rabbit gallbladder epithelium by Percoll density gradient centrifugation

The apical membranes of rabbit gallbladder epithelial cells were isolated by treating the homogenate with Ca2+ or Mg2+ and centrifuging the suspension in Percoll gradient. In this way brush-border membranes were obtained with enrichment factors ranging between 10 and 20 and yields of 15-30%. A second method is described with which membranes were isolated, without any preliminary treatment, first by differential centrifugation, then with Percoll gradient; the final membrane enrichment was over 15, however the yield was very low (3%). Many possible enzymatic markers of the apical plasma membrane were investigated: L-gamma-glutamyltransferase, alkaline phosphatase, leucine aminopeptidase, sucrase. The first appears to be that of choice. Apical membrane fraction could be also evidenced by autofluorescence or by labeling with Lotus tetragonolobus lectin. Preliminary experiments showed that apical plasma membranes isolated in this way form vesicles.     

Observations on the germinability in vitro of pollen from Camellia japonica L. (Theaceae)

Pollen grains from samples of Camellia japonica living in soil (A) and in greenhouse (B) were collected daily from just-opened anthers. The pollen was sown in various liquid media and incubated at 28 C. Higher germinative ability of A in comparison with B was observed and related to the relative humidity which is higher in soil than in greenhouse. This phenomenon recurs even if the composition of the culture medium is changed. However the composition of the medium greatly influences the percentage of germination of both A and B notwithstanding the environmental conditions. The pollen collected from anthers dehiscing in the first day of the anthesis has a high germinative ability which suddenly decreases in the ripe pollen of the following days.