Understanding the metabolic burden of recombinant antibody production in Saccharomyces cerevisiae using a quantitative metabolomics approach

The cellular changes induced by heterologous protein expression in the yeast Saccharomyces cerevisiae have been analysed on many levels and found to be significant. However, even though high‐level protein production poses a metabolic burden, evaluation of the expression host at the level of the metabolome has often been neglected. We present a comparison of metabolite profiles of a wild‐type strain with those of three strains producing recombinant antibody variants of increasing size and complexity: an scFv fragment, an scFv–Fc fusion protein and a full‐length IgG molecule. Under producing conditions, all three recombinant strains showed a clear decrease in growth rate compared with the wild‐type strain and the severity of the growth phenotype increased with size of the protein. The levels of 76 intracellular metabolites were determined using a targeted (semi) quantitative mass spectrometry based approach. Based on unsupervised and supervised multivariate analysis of metabolite profiles, together with pathway activity profiling, the recombinant strains were found to be significantly different from each other and from the wild‐type strain. We observed the most prominent changes in metabolite levels for metabolites involved in amino acid and redox metabolism. Induction of the unfolded protein response was detected in all producing strains and is considered to be a contributing factor to the overall metabolic burden on the cells.     

Application of classification and regression trees for sensitivity analysis of the Escherichia coli O157:H7 food safety process risk model

Microbial food safety process risk models are simplifications of the real world that help risk managers in their efforts to mitigate food safety risks. An important tool in these risk assessment endeavors is sensitivity analysis, a systematic method used to quantify the effect of changes in input variables on model outputs. In this study, a novel sensitivity analysis method called classification and regression trees was applied to safety risk assessment with the use of portions of the Slaughter Module and Preparation Module of the E. coli O157:H7 microbial food safety process risk as an example. Specifically, the classification and regression trees sensitivity analysis method was evaluated on the basis of its ability to address typical characteristics of microbial food safety process risk models such as nonlinearities, interaction, thresholds, and categorical inputs. Moreover, this method was evaluated with respect to identification of high exposure scenarios and corresponding key inputs and critical limits. The results from the classification and regression trees analysis applied to the Slaughter Module confirmed that the process of chilling carcasses is a critical control point. The method identified a cutoff value of a 2.2-log increase in the number of organisms during chilling as a critical value above which high levels of contamination would be expected. When classification and regression trees analysis was applied to the cooking effects part of the Preparation Module, cooking temperature was found to be the most sensitive input, with precooking treatment (i.e., raw product storage conditions) ranked second in importance. This case study demonstrates the capabilities of classification and regression trees analysis as an alternative to other statistically based sensitivity analysis methods, and one that can readily address specific characteristics that are common in microbial food safety process risk models.     

Risk of Vibrio transmission linked to the consumption of crustaceans in coastal towns of Côte d'Ivoire

The purpose of this study was to assess the risk of Vibrio spp. transmission from crustaceans to humans in two coastal towns of C?te d'Ivoire. Bacteriologic analysis was performed on 322 crustacean samples obtained from six markets in Abidjan and one in Dabou. Suspected Vibrio colonies were identified by morphological, cultural, biochemical, and molecular tests and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. PCR assays were used to further characterize Vibrio strains. A survey on consumption of crustaceans was conducted among 120 randomly selected households in Abidjan. Overall, Vibrio spp. were isolated from 7.8% of the crustacean samples studied, at levels as high as 6.3 log CFU/g. Of the Vibrio strains identified, 40% were V. alginolyticus, 36% were V. parahaemolyticus, and 24% were nontoxigenic V. cholerae; the latter two species can cause mild to severe forms of seafood-associated gastroenteritis. Among interviewed households, 11.7% reported daily consumption of crustaceans, confirming the high probability of exposure of human population to Vibrio spp., and 7.5% reported symptoms of food poisoning after consumption of crustaceans. The absence of genes encoding major virulence factors in the studied strains, i.e., cholera toxin (ctxA and ctxB) in V. cholerae and thermostable direct hemolysin (tdh) and thermostable direct hemolysin-related hemolysin (trh) in V. parahaemolyticus, does not exclude the possibility of exposure to pathogenic strains. However, human infections are not common because most households (96.7%) boil crustaceans, usually for at least 45 min (85.9% of households) before consumption.     

Lycopene from two food sources does not affect antioxidant or cholesterol status of middle-aged adults

Background

Epidemiological studies have reported associations between reduced cardiovascular disease and diets rich in tomato and/or lycopene. Intervention studies have shown that lycopene-containing foods may reduce cholesterol levels and lipid peroxidation, factors implicated in the initiation of cardiovascular disease. The objective of this study was to determine whether consumption of lycopene rich foods conferred cardiovascular protection to middle-aged adults as indicated by plasma lipid concentrations and measures of ex vivo antioxidants.

Methods

Ten healthy men and women consumed a low lycopene diet with no added lycopene (control treatment) or supplemented with watermelon or tomato juice each containing 20 mg lycopene. Subjects consumed each treatment for three weeks in a crossover design. Plasma, collected weekly was analyzed for total cholesterol, high density lipoprotein cholesterol (HDL-C) and triglyceride concentrations and for the antioxidant biomarkers of malondialdehyde formation products (MDA), plasma glutathione peroxidase (GPX) and ferric reducing ability of plasma (FRAP). Data were analyzed using Proc Mixed Procedure and associations between antioxidant and lipid measures were identified by Pearson's product moment correlation analysis.

Results

Compared to the control diet, the lycopene-containing foods did not affect plasma lipid concentrations or antioxidant biomarkers. Women had higher total cholesterol, HDL-C and triglyceride concentrations than did the men. Total cholesterol was positively correlated to MDA and FRAP while HDL-C was positively correlated to MDA and GPX. GPX was negatively correlated to triglyceride concentration.

Conclusions

The inclusion of watermelon or tomato juice containing 20 mg lycopene did not affect plasma lipid concentrations or antioxidant status of healthy subjects. However, plasma cholesterol levels impacted the results of MDA and FRAP antioxidant tests.     

有限变形理论能量释放率增率形式

基于有限变形理论中的能量原理和变分原理,考虑以有裂纹的瞬时位形为参考,建立增量变形引起的裂纹扩展方程能够更真实的描述裂纹尖端的扩展机制,在含有裂纹物体的瞬时变形的基础上,推导了裂纹体的能量释放率和增率的形式,提出了裂纹扩展判据.该判据反映了单位时间内裂纹扩展单位面积可以提供的能量与单位时间内裂纹扩展单位面积所需要的能量...     

In situ FT-IR measurements of competitive vapor adsorption into porous thin films containing silica nanoparticles

Vapor adsorption into porous ultrathin films on a gold surface is investigated with in situ surface plasmon resonance (SPR) and polarization-modulation infrared reflection-absorption spectroscopy (PM-IRRAS). The thin films are prepared by the electrostatic self-assembly of oppositely charged poly(L-lysine) (PL) and silica nanoparticles on a chemically modified gold surface. Characterization with ex situ SPR and PM-IRRAS demonstrates the buildup of multiple PL/SiO2 bilayers as well as an excellent correlation between the quantitative results from these two techniques. In situ vapor adsorption experiments with these thin films show evidence of porosity, reproducibility, and rapid reversibility. Exposure to acetone vapor (P/P0 = 0.032) causes the film to adsorb 9% acetone by volume, which corresponds to coverage of approximately one-half of the silica nanoparticle surface area. In situ PM-IRRAS provides much information about the molecular interactions occurring in the film upon adsorption or desorption of vapors. Dosing with a mixture of vapors leads to a competition for adsorption into the film, and PM-IRRAS results show that acetone slightly outcompetes nitromethane. These experiments with nanoparticle thin films demonstrate the advantages of using in situ PM-IRRAS for studying reversible adsorption in the presence of vapor mixtures.     

Impedance criterion for power modules comparison

It is well known that power modules must exhibit as low as possible stray inductance. However, the total inductance of a power module is not the only parameter to indicate the electrical quality of the packaging. In this paper, mathematical criteria will be given, which will allow the evaluation of power modules interconnects with regard to several objectives: equal current constraints among the different paralleled dies, power-drive, and even drive-drive interactions.     

Analysis of injection-locked gain-guided diode laser arrays

A new model for injection-locked gain guided laser arrays is proposed. Diffraction-limited and single-lobe operation of injection-locked arrays is attributed to coherent summation of several transverse modes that are phase locked by injection. The model predicts far-field and near-field patterns, locking bandwidth, beam-steering properties, and locked output power. The effects of varying the master power, beam shape, position, and incidence angle on the slave array facet are also studied. Theoretical and experimental results are compared