Influence of pH and temperature on the growth of and toxin production by neurotoxigenic strains of Clostridium butyricum type E

Strains of Clostridium butyricum that produce botulinal toxin type E have been implicated in outbreaks of foodborne botulism in China, India, and Italy, yet the conditions that are favorable for the growth and toxinogenesis of these strains remain to be established. We attempted to determine the temperatures and pH levels that are most conducive to the growth of and toxin production by the six strains of neurotoxigenic C. butyricum that have been implicated in outbreaks of infective and foodborne botulism in Italy. The strains were cultured for 180 days on Trypticase-peptone-glucose-yeast extract broth at various pHs (4.6, 4.8, 5.0, 5.2, 5.4, 5.6, and 5.8) at 30 degrees C and at various temperatures (10, 12, and 15 degrees C) at pH 7.0. Growth was determined by checking for turbidity; toxin production was determined by the mouse bioassay. We also inoculated two foods: mascarpone cheese incubated at 25 and 15 degrees C and pesto sauce incubated at 25 degrees C. The lowest pH at which growth and toxin production occurred was 4.8 at 43 and 44 days of incubation, respectively. The lowest temperature at which growth and toxin production occurred was 12 degrees C, with growth and toxin production first being observed after 15 days. For both foods, toxin production was observed after 5 days at 25 degrees C. Since the strains did not show particularly psychrotrophic behavior, 4 degrees C can be considered a sufficiently low temperature for the inhibition of growth. However, the observation of toxin production in foods at room temperature and at abused refrigeration temperatures demands that these strains be considered a new risk for the food industry.     

Nucleosome dynamics. III. Histone tail-dependent fluctuation of nucleosomes between open and closed DNA conformations. Implications for chromatin dynamics and the linking number paradox. A relaxation study of mononucleosomes on DNA minicircles

The mean linking number () of the topoisomer equilibrium distribution obtained upon relaxation of DNA minicircles with topoisomerase I did not increase linearly, but rather in a step wise fashion, with DNA size between 351 and 366 bp. As a consequence, the corresponding linking number difference () did not remain equal to 0, but rather oscillated between +/-0.3 with the periodicity of the double helix. This oscillation, not observed with plasmid-size DNA, is an expected consequence of the stiffness of short DNA. When minicircles were reconstituted with a nucleosome, the associated oscillated between approximately -1.4 +/-0. 2. This oscillation appears to result from the combined effects of DNA stiffness, and nucleosome ability to thermally fluctuate between three distinct DNA conformational states. Two of these states, a closed approximately 1.75-turn DNA conformation with negatively crossed entering and exiting DNAs, and an open approximately 1.4-turn conformation with uncrossed DNAs, are well known, whereas the third state, with a closed DNA conformation and DNAs tending to cross positively rather than negatively, is less familiar. Access to both closed "negative" and "positive" states appears to be mediated by histone N-terminal tails, as shown by specific alterations to the oscillation caused by histone acetylation and phosphate ions, a potent tail destabilizator. These results extend previous observations of ethidium bromide fluorescence titration in the accompanying article, which have pointed to an histone tail-dependent flexibility of entering and exiting DNAs to positive crossing. They also show that DNA wrapping around the histones occurred without twist alteration compared to the DNA free in solution, and reveal an intriguing new facet of the "linking-number-paradox" problem: the possibility for linkers in chromatin to adopt different crossing status within an overall dynamic equilibrium which may be regulated by histone acetylation.     

Efficient Decomposition Techniques for Symbolic Analysis of Large-Scale Analog Circuits by State Variable Method

New tearing techniques for the systematic formulation of the state equations in symbolic normal-form for linear and nonlinear time-invariant large-scale analog circuits are developed. Some examples are given to illustrate the decomposition procedure and the assignment of the connection sources. The partial symbolic state equations of the opamp A 741 are obtained using the tearing approach, while these equations cannot be written working on the whole circuit.     

Influence of air abrasion preparation on microleakage in glass ionomer cement restorations

The aim of this study was to assess microleakage in class V cavities prepared by air abrasion or high-speed dental bur and restored with different glass ionomer cements. Sixty bovine incisors were equally divided into 6 groups: I, II and III (preparation by high-speed) and IV, V and VI (preparation by air abrasion). Groups I and IV were restored with Fuji IX; groups II and V with Ketac Molar; and groups III and VI with Vitremer. After 24 h (37 °C), specimens were thermocycled, isolated with nail varnish, immersed in a 0.2% Rhodamine B solution for 24 hours, sectioned longitudinally and analyzed for microleakage using an optical microscope connected to a digital camera and a computer. The images were digitized and a software allowed the quantitative evaluation of microleakage in millimeters. Data were analyzed by Wilcoxon and Kruskal-Wallis tests. It was observed that there were significant differences (p ≤ 0.05) between incisal (enamel) and cervical (dentine/cementum) margins, mainly for Ketac Molar; there was no difference (p ≥ 0.05) between preparation methods, except for group II (high-speed/Ketac Molar) that showed higher infiltration; regarding the materials, Ketac Molar demonstrated the highest microleakage values (p ≤ 0.05), and only Vitremer sealed completely both margins of restorations. It was concluded that air abrasion preparation did not influence microleakage in class V restorations with the employed glass ionomer cements.     

Hourly indoor radon measurements in a research house

This paper reports and discusses the behaviour of radon concentration with time in an uninhabited dwelling. The relationship between variations in radon concentrations and indoor-outdoor temperatures and wind intensity has also been discussed. Radon concentration was measured hourly in a house located at a height of 800 m in the Lombard Prealps, at the top of the Valassina valley. The wind velocity and indoor-outdoor temperatures were measured by means of a meteorological station located on the terrace of the house. The data were analysed using the LBL model for indoor-outdoor air exchange and the models for the indoor accumulation of radon due to exhalation from building materials and pressure-driven infiltrations located underground. The role of wind and indoor-outdoor temperatures were analysed. The agreement of measurements with modelling clearly demonstrates the importance of the different sources of indoor radon. As the investigation was conducted in an uninhabited house, the measurements were not affected by the behaviour of people, e.g. opening and closing of windows. Measurements of the outdoor atmospheric concentrations of (222)Rn provide an index of the atmospheric stability, the formation of thermal inversions and convective turbulence.     

Laser-induced breakdown spectroscopy analysis of energetic materials

A number of energetic materials and explosives have been studied by laser-induced breakdown spectroscopy (LIBS). They include black powder, neat explosives such as TNT, PETN, HMX, and RDX (in various forms), propellants such as M43 and JA2, and military explosives such as C4 and LX-14. Each of these materials gives a unique spectrum, and generally the spectra are reproducible shot to shot. We observed that the laser-produced microplasma did not initiate any of the energetic materials studied. Extensive studies of black powder and its ingredients by use of a reference spectral library have demonstrated excellent accuracy for unknown identification. Finally, we observed that these nitrogen- and oxygen-rich materials yield LIBS spectra in air that have correspondingly different O:N peak ratios compared with air. This difference can help in the detection and identification of such energetic materials.     

A homology model of penicillin acylase from Alcaligenes faecalis and in silico evaluation of its selectivity

A three-dimensional model of the relatively unknown penicillin acylase from Alcaligenes faecalis (PA-AF) was built up by means of homology modeling based on three different crystal structures of penicillin acylase from various sources. An in silico selectivity study was performed to compare this homology model to the structure of the Escherichia coli enzyme (PA-EC) in order to find any selectivity differences between the two enzymes. The program GRID was applied in combination with the principal component analysis technique to identify the regions of the active sites where the PAs potentially engage different interactions with ligands. These differences were further analyzed and confirmed by molecular docking simulations. The PA-AF homology model provided the structural basis for the explanation of the different enantioselectivities of the enzymes previously demonstrated experimentally and reported in the literature. Different substrate selectivities were also predicted for PA-AF compared to PA-EC. Since no crystallographic data are available for PA-AF to date, the three-dimensional homology model represents a useful and efficient tool for fully exploiting this attractive and efficient biocatalyst, particularly in enantioselective acylations of amines.     

CSL model checking algorithms for QBDs

We present an in-depth treatment of model checking algorithms for a class of infinite-state continuous-time Markov chains known as quasi-birth death processes. The model class is described in detail, as well as the logic CSL to express properties of interest. Using a new property-independency concept, we provide model checking algorithms for all the CSL operators. Special emphasis is given to the time-bounded until operator for which we present a new and efficient computational procedure named uniformization with representatives. By the use of an application-driven dynamic stopping criterion, the algorithm stops whenever the property to be checked can be certified (or falsified). A comprehensive case study of a connection management system shows the versatility of our new algorithms.     

An application-centric evaluation of OpenCL on multi-core CPUs

Although designed as a cross-platform parallel programming model, OpenCL remains mainly used for GPU programming. Nevertheless, a large amount of applications are parallelized, implemented, and eventually optimized in OpenCL. Thus, in this paper, we focus on the potential that these parallel applications have to exploit the performance of multi-core CPUs. Specifically, we analyze the method to systematically reuse and adapt the OpenCL code from GPUs to CPUs. We claim that this work is a necessary step for enabling inter-platform performance portability in OpenCL.     

Reference spectra to classify Amazon water types

Reference spectra extracted from spectral libraries can distinguish different water types in images when associated with limnological information. In this study, we compiled available databases into a single spectral library, using field water reflectance spectra and limnological data collected by different researchers and campaigns in the Amazonian region. By using an iterative clustering procedure based on the combination of reflectance and optically active components (OACs), reference spectra representative of the major Amazonian water types were defined from this library. Differences between the resultant limnological classes were also evaluated by paired t-tests at significance level 0.05. Finally, reference spectra were tested for Spectral Angle Mapper (SAM) classification of waters in Hyperion/Earth Observing-One (EO-1) and Medium Resolution Imaging Spectrometer (MERIS)/Environment Satellite (Envisat) images acquired simultaneously as the field campaigns. Results showed highly variable concentrations of OACs due to the complexity of the Amazonian aquatic environments. Ten classes were defined to represent this complexity, broadly grouped into four limnological characteristics: clear waters with low concentrations of OACs (class 1); black waters rich in dissolved organic carbon (DOC) (class 2); waters with large concentrations of inorganic suspended solids (ISSs) (classes 3–7); and waters dominated by chlorophyll-a (chl-a) (classes 8–10). Using the ten reference spectra, SAM classification of the field water curves produced an overall accuracy of 86% with the highest values observed for classes 3, 4, 6 and 7 and the lowest accuracy for classes 1 and 2. The results of paired t-tests confirmed the class differences based on the concentrations of OACs. SAM classification of the Hyperion and MERIS images using ground truth information resulted in overall classification accuracies of 48% and 67%, respectively, with the highest errors associated with specific portions of the scenes that were not adequately represented in the spectral library.