New Insights into Bacterial Chemoreceptor Array Structure and Assembly from Electron Cryotomography

Bacterial chemoreceptors cluster in highly ordered, cooperative, extended arrays with a conserved architecture, but the principles that govern array assembly remain unclear. Here we show images of cellular arrays as well as selected chemoreceptor complexes reconstituted in vitro that reveal new principles of array structure and assembly. First, in every case, receptors clustered in a trimers-of-dimers configuration, suggesting this is a highly favored fundamental building block. Second, these trimers-of-receptor dimers exhibited great versatility in the kinds of contacts they formed with each other and with other components of the signaling pathway, although only one architectural type occurred in native arrays. Third, the membrane, while it likely accelerates the formation of arrays, was neither necessary nor sufficient for lattice formation. Molecular crowding substituted for the stabilizing effect of the membrane and allowed cytoplasmic receptor fragments to form sandwiched lattices that strongly resemble the cytoplasmic chemoreceptor arrays found in some bacterial species. Finally, the effective determinant of array structure seemed to be CheA and CheW, which formed a “superlattice” of alternating CheA-filled and CheA-empty rings that linked receptor trimers-of-dimer units into their native hexagonal lattice. While concomitant overexpression of receptors, CheA, and CheW yielded arrays with native spacing, the CheA occupancy was lower and less ordered, suggesting that temporal and spatial coordination of gene expression driven by a single transcription factor may be vital for full order, or that array overgrowth may trigger a disassembly process. The results described here provide new insights into the assembly intermediates and assembly mechanism of this massive macromolecular complex

Mesoporous carbon-containing voltammetric biosensor for determination of tyramine in food products

A voltammetric biosensor based on tyrosinase (TYR) was developed for determination of tyramine. Carbon material (multi-walled carbon nanotubes or mesoporous carbon CMK-3-type), polycationic polymer—i.e., poly(diallyldimethylammonium chloride) (PDDA), and Nafion were incorporated into titania dioxide sol (TiO(2)) to create an immobilization matrix. The features of the formed matrix were studied by scanning electron microscopy (SEM) and cyclic voltammetry (CV). The analytical performance of the developed biosensor was evaluated with respect to linear range, sensitivity, limit of detection, long-term stability, repeatability, and reproducibility. The biosensor exhibited electrocatalytic activity toward tyramine oxidation within a linear range from 6 to 130 μM, high sensitivity of 486 μA mM(−1) cm(−2), and limit of detection of 1.5 μM. The apparent Michaelis–Menten constant was calculated to be 66.0 μM indicating a high biological affinity of the developed biosensor for tyramine. Furthermore, its usefulness in determination of tyramine in food product samples was also verified. [Figure: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00216-016-9612-y) contains supplementary material, which is available to authorized users

How portuguese and american teachers plan for literacy instruction

This study explored American and Portuguese elementary teachers' preferences in planning for literacy instruction using the Language Arts Activity Grid (LAAG; Cunningham, Zibulsky, Stanovich, & Stanovich, 2009), on which teachers described their preferred instructional activities for a hypothetical 2-h language arts block. Portuguese teachers (N = 186) completed Portuguese versions of a background questionnaire and LAAG electronically, in Survey Monkey; American teachers (N = 102) completed identical English measures using paper and pencil. Results showed that teachers in both groups usually addressed comprehension and reading fluency on their LAAGs and that they also allocated the most time to these two areas. However, American teachers were more likely to include teacher-directed fluency activities, whereas Portuguese teachers were more likely to include fluency activities that were not teacher directed. Significantly more American than Portuguese teachers addressed phonics in their planning, whereas significantly more Portuguese than American teachers addressed writing processes such as revision. Both groups of educators demonstrated large variability in planning, with many teachers omitting important components of literacy identified by researchers, for writing as well as reading. The study highlights the importance of providing teachers with comprehensive, research-based core literacy curricula as well as professional development on key components of literacy. Study findings also suggest significant relationships between orthographic transparency and teachers' instructional planning.This research was supported by a 2-year grant from the Foundation Francisco Manuel dos Santos in Portugal as well as by a Connecticut State University research grant in the U.S.A. We would like to express our sincere gratitude to these funding agencies as well as to the teachers and school districts who participated in the study and sent messages of interest about our research. In addition, warm thanks to our research assistants for their help with data collection, coding, and analysis, and to Anne Cunningham for providing us with inspiration as well as guidance in this work.info:eu-repo/semantics/publishedVersio

Wpływ domieszki napowietrzającej i rodzaju cementu na właściwości świeżej zaprawy

In order to explain the effect of air entraining admixture (AEA) and the type of cement on the properties of fresh mortar, the following tests were carried: bulk density, flaw and air content of 20 air entrained mortars and 20 nan air entrained mortars made of ordinary Portland cement (OPC) and blended Portland cements: blast furnace slag (BSF) - limestone powder (LP), fly ash - limestone, fly ash (FA) and (BFS) cement. It has been found that the modern anionic AEAs from the group of alkyl sulfates are much more efficient and active than traditional ones i.e. sodium abietate. It is difficult to entrain air into mortars with cement of various types to the same content of air. This requires to carry out a number of tests to determine the appropriate amounts of AEA. The air entrainment of LP cement mortar does not significantly change its flow. The air entrainment of BSF and FA cement mortars increases their workability. The lowest content of air was stated in non-air-entrained mortars with FA cement, the highest air content was observed in BFS cement mortars. According to the results the bulk density of mortars is inversely proportional to the amount of air entrained

The effect of coarse aggregate on the deformation properties of normal concretes

W artykule przedstawiono wyniki badań betonów zwykłych o wartościach w/c wynoszących 0,70 i 0,58. Badano odkształcalność doraźną betonów w zakresie modułu sprężystości, współczynnika odkształcenia poprzecznego oraz wartości odkształceń granicznych przy maksymalnym naprężeniu. Ponadto przedstawiono przebieg krzywych sigma-epsilon w pełnym zakresie obciążenia. Czynnikiem zmiennym w badaniach było kruszywo grube (grys bazaltowy, granitowy, dolomitowy oraz dla porównania kruszywo naturalne otoczakowe). W obydwu grupach betonów (o tym samym w/c) stwierdzono istotny wpływ rodzaju kruszywa grubego na badane cechy odkształceniowe betonów zwykłych.The test results presented concern normal concretes of the w/c-ratio 0.58 and 0.70. There have been examined a modulus of elasticity, transverse strain coefficient and the strain at the highest compressive stress. There are presented the curves sigma-epsilon for the whole range of loading. The variable factor in the studies is the kind of coarse aggregate (crushed basalt, granite and dolomite compared with the natural gravel). There is stated a significant influence of aggregate on the deformation properties for both concrete groups of the same w/c-ratio