Application of direct plaque assay for detection and enumeration of bacteriophages of Bacteroides fragilis from contaminated-water samples

The direct double-agar-layer plaque assay for the detection and enumeration of specific bacteriophages of Bacteroides fragilis from contaminated-water samples was performed. Several factors that affect the methods, such as conditions of the bacterial culture, composition of the assay medium, addition of divalent cations, and decontamination techniques applied to the sample, were evaluated. The results obtained show that the direct assay technique proved to be more efficient than the most-probable-number technique. A higher recovery of bacteriophages was obtained from 17 of 24 samples with the direct assay. The two methods only showed similar results from samples with a low degree of pollution.     

Characterization of the Pre-meiotic S Phase through Incorporation of BrdU during Spermatogenesis in the Rat

Seminiferous tubules in mammals have histological arrangements defined by the associations between somatic cells and germ cells. The processes of DNA synthesis in meiotic and mitotic cells have different features that are not easily distinguishable through morphological means. In order to characterize the pre-meiotic S phase, 5-bromo-2’-deoxyuridine (BrdU) was injected intraperitoneally into Wistar rats, which were sacrificed 30 min, 2 hr, and 24 hr after injection. We found three different labeling patterns. One of these patterns was characterized by a distribution of the label in the form of speckles, most of which were associated with the nuclear envelope (labeling type I). We suggest that this pattern is due to mitotic DNA synthesis of type B spermatogonia. Labeling type II consisted of labeled foci scattered throughout the nuclear volume, which can be correlated with preleptotenic cells in pre-meiotic DNA synthesis. After 24 hr of incorporation, a third type of labeling, characterized by large speckles, was found to be related to cells in the “bouquet” stage; that is, cells in transition between the leptotene and zygotene phases. Our results indicate that BrdU incorporation induces different labeling patterns in the mitotic and pre-meiotic S phases and thus makes it possible to identify somatic and germinal cells.     

Species formation and geographical range evolution in a genus of Central American cloud forest salamanders (Dendrotriton)

Aim Montane Central America offers an ideal system for testing geographical hypotheses of species diversification. We examined how the complex geological history of Nuclear Central America has shaped the diversification of a genus of cloud‐forest‐inhabiting salamanders (Dendrotriton). We applied parametric models of geographical range evolution to determine the predominant mode of species formation within the genus and to test existing hypotheses of geographical species formation in the region. Location Montane cloud forests of Nuclear Central America. Methods We estimated a species tree for Dendrotriton using a multi‐locus DNA sequence data set and several coalescent methods, and performed molecular dating for divergence events within the genus. We then applied the species‐tree estimate to a likelihood‐based time‐stratified model of geographical range evolution, based on current species distributions and available geological information for Central America. Results Species trees from all methods contain two groups, one corresponding to species from the Sierra de los Cuchumatanes and the other containing all remaining species. In most cases, species formation within the genus involved an even division of the geographical range of the ancestral species between descendant species. The ancestor of extant Dendrotriton species was estimated to have occurred in either the Sierra de los Cuchumatanes or the Sierra Madre de Chiapas, and both of these areas appear to have been important for diversification within the genus. The single species found in the Quaternary‐age Guatemalan volcanic cordillera dispersed to the volcanoes from an older highland area. Main conclusions Models of geographical range evolution, when combined with robust species‐tree estimates, provide insight into the historical biogeography of taxa not available from phylogenies or distributional data alone. Vicariant species formation, rather than peripatric or gradient speciation, appears to have been the dominant process of diversification, with most divergence events occurring within or between ancient highland areas. The apparent dispersal of Dendrotriton to the Quaternary‐age volcanoes raises the possibility that the rich salamander community there is composed of species that dispersed from geologically older areas. The Motagua Valley appears not to have been as important in vicariant species formation within Dendrotriton as it is within other groups.     

The solvation of bromide anion in acetonitrile: a structural study based on the combination of theoretical calculations and X-ray absorption spectroscopy

This work studies the solvation of bromide in acetonitrile by combining quantum mechanics, computer simulations and X-ray absorption near edge structure (XANES) spectroscopy. Three different sets of interaction potentials were tested, one of them derived from literature and the other two are simple modifications of the previous one to include specificities of the bromide–acetonitrile interactions. Results for microsolvation of bromide were obtained by quantum mechanical optimization and classical minimization of small clusters [Br(ACN) _n ]^?  (n = 9, 20). Analysis of molecular dynamics (MD) simulations has provided structural, dynamic and energetic aspects of the solvation phenomenon. The theoretical computation of Br K-edge XANES spectrum in solution using the structural information obtained from the different simulations has allowed the comparison among the three different potentials, as well as the examination of the main structural and dynamic factors determining the shape of the experimental spectrum.     

Simultaneous Detection of Fenitrothion and Chlorpyrifos-Methyl with a Photonic Suspension Array

A technique was developed for simultaneous detection of fenitrothion (FNT) and chlorpyrifos-methyl (CLT) using a photonic suspension array based on silica colloidal crystal beads (SCCBs). The SCCBs were encoded with the characteristic reflection peak originating from the stop-band of colloidal crystal. This approach avoids the bleaching, fading or potential interference seen when encoding by fluorescence. SCCBs with a nanopatterned surface had increased biomolecule binding capacity and improved stability. Under optimal conditions, the proposed suspension array allowed simultaneous detection of the selected pesticides in the ranges of 0.25 to 1024 ng/mL and 0.40 to 735.37 ng/mL, with the limits of detection (LODs) of 0.25 and 0.40 ng/mL, respectively. The suspension array was specific and had no significant cross-reactivity with other chemicals. The mean recoveries in tests in which samples were spiked with target standards were 82.35% to 109.90% with a standard deviation within 9.93% for CLT and 81.64% to 108.10% with a standard deviation within 8.82% for FNT. The proposed method shows a potentially powerful capability for fast quantitative analysis of pesticide residues.     

Modular synthesis of Petri nets for modeling flexible manufacturing systems

This paper proposes a modular Petri net synthesis method for modeling flexible manufacturing systems based on synchronization among control processes of the manufacturing resources (such as robots and machines). In the method, the target system is modeled in a bottom-up and uniform manner by first describing the system's control processes using strongly connected state machines (SCSMs) as the basic modules. Each SCSM may contain multiple tokens to represent resources from the same type such as spaces in a buffer. Next, the common transitions and common transition subnets of the modules are merged to represent their synchronization. The system model constructed is proven to be conservative and thus bounded. Moreover, a restricted class of merged nets is proven to be live and reversible. For general classes of merged nets, this paper shows theorems that easily calculateP-invariants of the final net without solving the linear system equations. TheseP-invariants can be used to help in verifying the model's qualitative properties such as liveness.     

An immunoenzymatic method for improving the sensitivity of antigen measurement by electroimmunodiffusion techniques. Application to the quantification of human alpha-fetoprotein.

A double antibody technique of electroimmunodiffusion, which uses glucose oxidase-labelled sheep antibodies to rabbit immunoglobulins as second antibody, is described. Primary antigen-antibody reaction is carried out with a rabbit antiserum by electroimmundiffusion. The glucose oxidase-labelled immunoreagent, being of general application, can serve for the quantification of different antigens and is here used for measurement of low levels of human alpha-fetoprotein. Reproducible results in the range of 50-800 ng/ml were obtained with a variation coefficient of 5 to 10%.