Thermal denaturation and aggregation of Myosin subfragment 1 isoforms with different essential light chains

We compared thermally induced denaturation and aggregation of two isoforms of the isolated myosin head (myosin subfragment 1, S1) containing different "essential" (or "alkali") light chains, A1 or A2. We applied differential scanning calorimetry (DSC) to investigate the domain structure of these two S1 isoforms. For this purpose, a special calorimetric approach was developed to analyze the DSC profiles of irreversibly denaturing multidomain proteins. Using this approach, we revealed two calorimetric domains in the S1 molecule, the more thermostable domain denaturing in two steps. Comparing the DSC data with temperature dependences of intrinsic fluorescence parameters and S1 ATPase inactivation, we have identified these two calorimetric domains as motor domain and regulatory domain of the myosin head, the motor domain being more thermostable. Some difference between the two S1 isoforms was only revealed by DSC in thermal denaturation of the regulatory domain. We also applied dynamic light scattering (DLS) to analyze the aggregation of S1 isoforms induced by their thermal denaturation. We have found no appreciable difference between these S1 isoforms in their aggregation properties under ionic strength conditions close to those in the muscle fiber (in the presence of 100 mM KCl). Under these conditions kinetics of this process was independent of protein concentration, and the aggregation rate was limited by irreversible denaturation of the S1 motor domain.     

High Catalytic Activity in CO Oxidation over MnO x Nanocrystals

Manganese oxides of various stoichiometry were prepared via Mn-oxalate precipitation followed by thermal decomposition in the presence of oxygen. A non-stoichiometric manganese oxide, MnO _x (x = 1.61…1.67) was obtained by annealing at 633 K and demonstrated superior CO oxidation activity, i.e. full CO conversion at room temperature and below. The activity gradually decreased with time-on-stream of the reactants but could be easily recovered by heating at 633 K in the presence of oxygen. CO oxidation over MnO _x in the absence of oxygen proved to be possible with reduced rates and demonstrated a Mars—van Krevelen—type mechanism to be in operation. A TEM structural analysis showed the MnO _x phase to form microrods with large aspect ratio which broke up into nanocrystalline manganese oxide (MnO _x ) particles with diameters below 3 nm and a BET specific surface area of 525 m²/g. Annealing at 798 K rather than 633 K produced well crystalline Mn₂O₃ which showed lower CO oxidation activity, i.e. 100% CO conversion at 335 K. The catalytic performance in CO oxidation of various Mn-oxides either studied in this work or elsewhere was compared on the basis of specific reaction rates.     

Relationships between Bacteroides 16S rRNA genetic markers and presence of bacterial enteric pathogens and conventional fecal indicators

Occurrence and prevalence of different bacterial enteric pathogens as well as their relationships with conventional (total and fecal coliforms) and alternative fecal indicators (host-specific Bacteroides 16S rRNA genetic markers) were investigated for various water samples taken from different sites with different degrees of fecal contamination. The results showed that a wide range of bacterial pathogens could be detected in both municipal wastewater treatment plant samples and in surface water samples. Logistic regression analysis revealed that total and human-specific Bacteroides 16S rRNA genetic markers showed significant predictive values for the presence of Escheriachia coli O-157, Salmonella, heat-labile enterotoxin (LT) of enterotoxigenic E. coli (ETEC), and heat-stable enterotoxin for human (STh) of ETEC. The probability of occurrence of these pathogenic bacteria became significantly high when the concentrations of human-specific and total Bacteroides 16S rRNA genetic markers exceeded 10(3) and 10(4) copies/100 mL. In contrast, Clostridium perfringens was detected at high frequency regardless of sampling sites and levels of Bacteroides 16S rRNA genetic markers. No genes related to Shigella spp., Staphylococcus aureus and Vibrio cholerae were detected in any samples analyzed in this study. Conventional indicator microorganisms had low levels of correlation with the presence of pathogens as compared with the alternative fecal indicators. These results suggested that real-time PCR-based measurement of alternative Bacteroides 16S rRNA genetic markers was a rapid and sensitive tool to identify host-specific fecal pollution and probably associated bacterial pathogens. However, since one fecal indicator might not represent the relative abundance of all pathogenic bacteria, viruses and protozoa, combined application of alternative indicators with conventional ones could provide more comprehensive pictures of fecal contamination, its source and association with pathogenic microorganisms.     

Influence of treatment time and pulse frequency on Salmonella Enteritidis, Escherichia coli and Listeria monocytogenes populations inoculated in melon and watermelon juices treated by pulsed electric fields

Consumption of unpasteurized melon and watermelon juices has caused several disease outbreaks by pathogenic microorganisms worldwide. Pulsed electric field (PEF) has been recognized as a technology that may inactivate those bacteria present in fluid food products at low temperatures. Hence, PEF treatment at 35 kV/cm, 4 mus pulse duration in bipolar mode and square shape were applied on Salmonella Enteritidis, E. coli and L. monocytogenes populations inoculated in melon and watermelon juices without exceeding 40 degrees C outlet temperatures. Different levels of treatment time and pulse frequency were applied to evaluate their effects on these microorganisms. Treatment time was more influential than pulse frequency (P相似文献   

A review on tomato authenticity: quality control methods in conjunction with multivariate analysis (chemometrics)

Authenticity and traceability have been two of the most important issues in the food chain. Authenticity in particular, is closely related with both food quality and safety issues. Vegetables stand for a category of foods heavily affected by adulteration either in terms of geographic origin (national or international level) or production methods (organic or conventional production, fertilizers, pesticides, genetically modified vegetables). This review aims at addressing most of the currently applied methods for ensuring quality control of vegetables; a) instrumental: ion chromatography, high pressure liquid chromatography, atomic absorption spectrophotometry, electronic nose and mass spectroscopy and b) sensory analysis. The results of all the above mentioned methods were analyzed by means of multivariate analysis (principal component analysis, discriminant analysis, cluster analysis, canonical analysis, and factor analysis). All ensuing results and conclusions are summarized in eight comprehensive tables.     

Browning Inhibition in Fresh-cut'Fuji'Apple Slices by Natural Antibrowning Agents

ABSTRACT Response surface methodology was used to study the effect of antibrowning agents (4‐hexylresorci‐nol, glutathione, N‐acetylcysteine, and ascorbic acid) and storage time (14 d) on the color of minimally processed Fuji apples. The selected color parameters were L*,a*, b*, hue angle (h*), and color difference (ΔE*). Storage time had a significant effect on all the studied color parameters (P± 0.05). 4‐hexylresorcinol showed the most effective individual effect on keeping constant a*values (P± 0.0001). Besides, the interaction of N‐acetylcysteine/ glutathione was found to have a significant effect (P± 0.05) on maintaining a* values over time. On the other hand, individual treatment with N‐acetylcysteine in concentrations higher than 0.75% w/v may be used to preserve a*and h*.According to the F‐test, 4‐hexylresorcinol and N‐acetylcysteine (P± 0.05) displayed a significant individual effect on ΔE*, indicating that ΔE* decreased when increasing the concentration of these antibrowning agents. Nevertheless, color difference went down when 4‐hexylresorcinol concentration increased up to 0.5%, but higher concentrations of this agent led to an increase in ΔE* that indicates browning.     

The effect of red pigment on the amyloidization of yeast proteins

The intensity of amyloid-bound thioflavine T fluorescence was studied in crude lysates of yeast strains carrying mutations in the ADE1 or ADE2 genes and accumulating the red pigment (a result of polymerization of aminoimidazoleribotide), and in white isogenic strains--either adenine prototrophs or carrying mutations at the first stages of purine biosynthesis. We found that the red pigment leads to a drop of amyloid content. This result, along with the data on separation of protein polymers of white and red strains in PAGE, suggests that the red pigment inhibits amyloid fibril formation. The differences in transmission of the thioflavine T fluorescence pattern by cytoduction and in blot-hybridization of pellet proteins of red and white [PSI(+) ] strains with Sup35p antibodies confirmed this conclusion. Purified red pigment treatment also led to a decrease of fluorescence intensity of thioflavine T bound to insulin fibrils and to yeast pellet protein aggregates from [PSI(+) ] strains. This suggests red pigment interaction with amyloid fibrils. Comparison of pellet proteins from red and white isogenic strains separated by 2D-electrophoresis followed by MALDI analysis has allowed us to identify 48 pigment-dependent proteins. These proteins mostly belong to functional classes of chaperones and proteins involved in glucose metabolism, closely corresponding to prion-dependent proteins that we characterized previously. Also present were some proteins involved in stress response and proteolysis. We suppose that the red pigment acts by blocking certain sites on amyloid fibrils that, in some cases, can lead in vivo to interfere with their contacts with chaperones and the generation of prion seeds.     

An exact optimization approach for a transfer line reconfiguration problem

The paper deals with a transfer line reconfiguration problem. Such lines are made of machines (workstations) located in sequence and linked by a material handling device. Each machine can be equipped with several multi-spindle heads activated sequentially. Each spindle head executes a set of operations simultaneously. If new products have to be manufactured at the line or existing products are modified, then the line has to be reconfigured in order to meet new production requirements. The objective of such reconfiguration is to reduce the investment cost for new equipment by reusing optimally the existing facilities. A new mathematical model is suggested for this optimization problem. A case study is presented to demonstrate the use of the developed optimization model. The results of numerical experiments for 41 industrial test problems are also analyzed which show that up to 51 % investment savings can be obtained with this model.     

Lessening polygalacturonase activity in a commercial enzyme preparation by exposure to pulsed electric fields

The effect of high-intensity pulsed electric field (HIPEF) treatments on polygalacturonase (PG) activity in an aqueous solution of a commercial enzyme preparation was evaluated. HIPEF treatments reduced PG activity notably. Exponentially decaying pulses of 40 and 160 µs generated by a laboratory scale device were applied in a batch processing and bipolar mode. Electric fields ranged from 5.18 to 19.39 kV/cm. The number of pulses ranged up to 400. Temperature was always below 25 °C. Maximum inactivation of PG activity (98%) required 32.4 ms HIPEF treatment at 10.28 kV/cm. PG activity depleted exponentially because of HIPEF treatments. The first order rate constants ranged from 32 to 590 µs ^-1 and increased exponentially with electric field intensity. PG activity decreased exponentially with input electric energy density.