alpha-Amylase Production in Fed BatchCultivation of Bacillus caldolyticus: An Interpretation of FermentationCourseUsing 2-D Gel Electrophoresis

The conditions for increased production of thermostable a-amylase from Bacillus caldolyticus DSM 405 were investigated. Preliminary experiments in batch shake flasks led to an optimized initial cultivation medium. Shake flask experiments in extended-batch and in fed-batch mode of operation indicated that the a-amylase production was enhanced by continuous feeding of starch. The activity of the a-amylase with optimized initial medium in batch-operated shake flasks was 5.7 U mL–1 compared to 15.4U mL–1 in the extended-batch culture and 21 U mL–1 in fed-batch culture. The improvements were achieved by avoiding any excess of starch in medium that led to accumulation of glucose followed by acetate formation. Adding casitone as the second component of the feeding solution in an aerated and agitated fed-batch bioreactor (3-liter working volume) led to an increased -amylase activity of up to 163.7 U mL.–1 All phases of cultivation were analyzed using 2D-gel electrophoresis in combination with nano LC-ESI-MS/MS for identification of altered proteins. Pyruvate kinase, 6-phosphofructokinase, GltC, anti-sigma F factor, glycogen synthase and several important variable proteins were detected. With help of these results, potential improvements of a two-component feeding strategy are discussed

Scaling in a continuous time model for biological aging

In this paper we consider a generalization to the asexual version of the Penna model for biological aging, where we take a continuous time limit. The genotype associated to each individual is an interval of real numbers over which Dirac $\delta$--functions are defined, representing genetically programmed diseases to be switched on at defined ages of the individual life. We discuss two different continuous limits for the evolution equation and two different mutation protocols, to be implemented during reproduction. Exact stationary solutions are obtained and scaling properties are discussed.Comment: 10 pages, 6 figure

Assessing agreement between preclinical magnetic resonance imaging and histology: An evaluation of their image qualities and quantitative results

One consequence of demographic change is the increasing demand for biocompatible materials for use in implants and prostheses. This is accompanied by a growing number of experimental animals because the interactions between new biomaterials and its host tissue have to be investigated. To evaluate novel materials and engineered tissues the use of nondestructive imaging modalities have been identified as a strategic priority. This provides the opportunity for studying interactions repeatedly with individual animals, along with the advantages of reduced biological variability and decreased number of laboratory animals. However, histological techniques are still the golden standard in preclinical biomaterial research. The present article demonstrates a detailed method comparison between histology and magnetic resonance imaging. This includes the presentation of their image qualities as well as the detailed statistical analysis for assessing agreement between quantitative measures. Exemplarily, the bony ingrowth of tissue engineered bone substitutes for treatment of a cleft-like maxillary bone defect has been evaluated. By using a graphical concordance analysis the mean difference between MRI results and histomorphometrical measures has been examined. The analysis revealed a slightly but significant bias in the case of the bone volume ðbiasHisto MRI: Bonevolume = 2: 40 %, p < 0: 005) and a clearly significant deviation for the remaining defect width ðbiasHisto MRI: Defectwidth = 6: 73 %, p 0: 005Þ: But the study although showed a considerable effect of the analyzed section position to the quantitative result. It could be proven, that the bias of the data sets was less originated due to the imaging modalities, but mainly on the evaluation of different slice positions. The article demonstrated that method comparisons not always need the use of an independent animal study, additionally

On the importance of hydrodynamic interactions in polyelectrolyte electrophoresis

The effect of hydrodynamic interactions on the free-solution electrophoresis of polyelectrolytes is investigated with coarse-grained molecular dynamics simulations. By comparing the results to simulations with switched-off hydrodynamic interactions, we demonstrate their importance in modelling the experimentally observed behaviour. In order to quantify the hydrodynamic interactions between the polyelectrolyte and the solution, we present a novel way to estimate its effective charge. We obtain an effective friction that is different from the hydrodynamic friction obtained from diffusion measurements. This effective friction is used to explain the constant electrophoretic mobility for longer chains. To further emphasize the importance of hydrodynamic interactions, we apply the model to end-labeled free-solution electrophoresis.Comment: 15 pages, 7 figures; accepted for publication in J. Phys.: Condens. Matte

Squalene production under oxygen limitation by Schizochytrium sp. S31 in different cultivation systems

Abstract: The triterpene squalene is widely used in the food, cosmetics and pharmaceutical industries due to its antioxidant, antistatic and anti-carcinogenic properties. It is usually obtained from the liver of deep sea sharks, which are facing extinction. Alternative production organisms are marine protists from the family Thraustochytriaceae, which produce and store large quantities of various lipids. Squalene accumulation in thraustochytrids is complex, as it is an intermediate in sterol biosynthesis. Its conversion to squalene 2,3-epoxide is the first step in sterol synthesis and is heavily oxygen dependent. Hence, the oxygen supply during cultivation was investigated in our study. In shake flask cultivations, a reduced oxygen supply led to increased squalene and decreased sterol contents and yields. Oxygen-limited conditions were applied to bioreactor scale, where squalene accumulation and growth of Schizochytrium sp. S31 was determined in batch, fed-batch and continuous cultivation. The highest dry matter (32.03 g/L) was obtained during fed-batch cultivation, whereas batch cultivation yielded the highest biomass productivity (0.2 g/L*h−1). Squalene accumulation benefited from keeping the microorganisms in the growth phase. Therefore, the highest squalene content of 39.67 ± 1.34 mg/g was achieved by continuous cultivation (D = 0.025 h−1) and the highest squalene yield of 1131 mg/L during fed-batch cultivation. Volumetric and specific squalene productivity both reached maxima in the continuous cultivation at D = 0.025 h−1 (6.94 ± 0.27 mg/L*h−1 and 1.00 ± 0.03 mg/g*h−1, respectively). Thus, the choice of a suitable cultivation method under oxygen-limiting conditions depends heavily on the process requirements. Key points: • Measurements of respiratory activity and backscatter light of thraustochytrids • Oxygen limitation increased squalene accumulation in Schizochytrium sp. S31 • Comparison of different cultivation methods under oxygen-limiting conditions

Splitting of Folded Strings in AdS_4*CP^3

We study classically splitting of two kinds of folded string solutions in AdS_4*CP^3. Conserved charges of the produced fragments are computed for each case. We find interesting patterns among these conserved charges.Comment: minor changes, 14 pages, no figure

Transfer Functions for Protein Signal Transduction: Application to a Model of Striatal Neural Plasticity

We present a novel formulation for biochemical reaction networks in the context of signal transduction. The model consists of input-output transfer functions, which are derived from differential equations, using stable equilibria. We select a set of 'source' species, which receive input signals. Signals are transmitted to all other species in the system (the 'target' species) with a specific delay and transmission strength. The delay is computed as the maximal reaction time until a stable equilibrium for the target species is reached, in the context of all other reactions in the system. The transmission strength is the concentration change of the target species. The computed input-output transfer functions can be stored in a matrix, fitted with parameters, and recalled to build discrete dynamical models. By separating reaction time and concentration we can greatly simplify the model, circumventing typical problems of complex dynamical systems. The transfer function transformation can be applied to mass-action kinetic models of signal transduction. The paper shows that this approach yields significant insight, while remaining an executable dynamical model for signal transduction. In particular we can deconstruct the complex system into local transfer functions between individual species. As an example, we examine modularity and signal integration using a published model of striatal neural plasticity. The modules that emerge correspond to a known biological distinction between calcium-dependent and cAMP-dependent pathways. We also found that overall interconnectedness depends on the magnitude of input, with high connectivity at low input and less connectivity at moderate to high input. This general result, which directly follows from the properties of individual transfer functions, contradicts notions of ubiquitous complexity by showing input-dependent signal transmission inactivation.Comment: 13 pages, 5 tables, 15 figure