Mechanism of simultaneous solutes separation and concentration by size exclusion cyclic separation

The semicontinuous separation/concentration of two solutes with different molecular size by the size exclusion cyclic separation method is based on the opposite swelling responses of two gels to a temperature change. Experimental results of separation and concentration of solutes are verified qualitatively by the theoretical models based on the local equilibrium assumption. Separation of two solutes is shown by the breakthrough curves in coupled gel columns. In closed coupled columns, the volumetric space for the large molecules which are totally excluded from the gels becomes smaller, creating a concentrating effect as the gels swell when temperature changes. A mechanistic model is suggested to predict the large molecule concentration to increase to its solubility limit as cycle repeats.     

Role of conserved Asn-Tyr-Asp-Tyr sequence in bacterial copper/2,4, 5-trihydroxyphenylalanyl quinone-containing histamine oxidase

Copper amine oxidase contains a covalently bound quinonoid cofactor, 2,4,5-trihydroxyphenylalanyl quinone (TPQ), which is synthesized by post-translational modification of a specific tyrosyl residue occurring in the highly conserved sequence, Asn-Tyr-(Asp/Glu)-Tyr. To elucidate the role(s) of the conserved sequence in the biogenesis of TPQ, each of the corresponding residues at positions 401-404 in the recombinant histamine oxidase from Arthrobacter globiformis has been replaced with other amino acids by site-directed mutagenesis. When Asn-401 was changed to Asp or Gln, the rate of TPQ formation by copper-dependent self-processing was 10(3)- to 10(4)-fold slower than in the wild-type enzyme. When Tyr-402 was replaced by Phe, TPQ was not formed at all, showing that Tyr-402 is essential as the precursor to TPQ. In contrast, Asp-403 could be replaced by Glu without changes in the rate of TPQ formation, whereas its replacement by Asn led to a marked decrease. Furthermore, when Tyr-404 was changed to Phe, TPQ was formed swiftly on incubation with copper ions, but the TPQ enzyme exhibited very low activity with altered substrate specificity. These results collectively indicate that a very rigorous structural motif is required for efficient formation of TPQ and for the catalytic activity in the active site of copper amine oxidases.     

Quantitative methods for scoring cell migration and invasion in filter-based assays

The ability of cells to traverse pores in a biocompatible filter provides means for examining cell chemoattraction. Filter-based assays also permit rapid, quantitative assessment of the in vitro migratory and invasive potential of tumor cells. Scoring migration has relied on visual counting of stained cells which appear on the underside of the filter and determining a true percentage score involves arduous counting of cells on both filter surfaces. Visual counting of random fields may be unreliable, and counting all fields is laborious. In the present study we developed and compared two alternative methods for scoring cell numbers in filter-based assays, a colorimetric assay of toluidine blue binding, and a radioassay of cells prelabeled with [3H]thymidine. Each method was evaluated for sensitivity, variability, ease of use and efficiency, and suitability for use in assays of cell migration and invasion. The radiolabeling method proved to be sensitive and reliable and was the most efficient technique. Although less sensitive and specific, the colorimetric dye method offered a rapid and reliable, nonradioactive alternative with the distinct advantage of preserving intact cultures for follow-up visual assessments. We conclude that colorimetric and radiolabel scoring of filter-based assays are reliable and efficient semiautomated methods which provide means to obtain more complete assessments of cell migration and invasion.     

Reduction of loading effect by tungsten etchback in a magneticallyenhanced reactive ion etcher

The plug loading effect occurring during the etchback of tungsten was investigated in a magnetically enhanced reactive ion etcher using SF ₆/Ar mixtures. It was found that while the plug loading effect is independent of varying SF₆/Ar flow rate ratio and magnetic field intensity, it is reduced under the condition of high selectivity of tungsten relative to TiN which was achieved at high chamber pressure and low RF power. It is proposed that when TiN is used as a glue layer, the W etch rate enhancement in the plug is mainly controlled by a local loading effect. Under the optimized etchback conditions the plug loss was successfully controlled without the tungsten residue left on severe topology     

Nursing diagnoses and construct validity of pain, self-care deficit and impaired mobility

Construct validity of the concepts of pain, self-care deficit and impaired mobility was examined in a study of 120 medical-surgical patients. Factors derived from a factor analysis of the empirical signs and symptoms were: pain-factor and self-care deficit/impaired mobility-factor. The pain-factor clearly showed discriminant validity by differentiating between those with and without Clinical Nurse Specialist (CNS)-identified diagnosis of pain. An overlap between the defining characteristics of the diagnoses of self-care deficit and impaired mobility has been suggested in the present sample of patients. Limitations of the study and recommendations for further research are described.     

Analysis of culture fluorescence by a fiber-optic sensor inNicotiana tabacum plant cell culture

The on-line sensing of viable cell weight in plant cell culture process is applied to analysis and control of process. The fiber-optic fluorescence sensor was constructed to measure the NADH-dependent fluorescence inNicotiana tabacum plant cell culture and the analysis of fluorescence signal was done to be correlated with the viable cell weight. The structured kinetic model for cell growth was proposed to estimate the theoretical viable cell weight. The dimensional analysis was proposed for the interpretation of fluorescence signal, in which the path length, the inner filter effect and the hydrodynamic conditions were considered as the key factors on fluorescence signal. The dimensional analysis and empirical correlation of fluorescence signal to viable cell weight was applied to the interpretation of the detected fluorescence signal during cultivation. The proposed interpretation of fluorescence signal using dimensional analysis was well correlated with the viable cell weight estimated by the structured kinetic model as well as by empirical correlation.     

Multiresolution segmentation of respiratory electromyographicsignals

Analysis of respiratory electromyographic (EMG) signals in the study of respiratory control requires the detection of burst activity from background (signal segmentation), and focuses upon the determination of onset and cessation points of the burst activity (boundary estimation). The authors describe a new automated multiresolution technique for signal segmentation and boundary estimation. During signal segmentation, a new transitional segment is defined which contains the boundary between background a burst activity. Boundary estimation is then performed within this transitional segment. Boundary candidates are selected and a probability is attributed to each candidate, using an artificial neural network. The final boundary for a given transitional segment is the boundary estimate with the maximum a posteriori probability. This new method has proved accurate when compared to boundaries chosen by two investigators     

The cue-familiarity heuristic in metacognition

Four experiments contrasted the cue-familiarity hypothesis of feeling-of-knowing judgments (FKJs) and tip-of-the-tongue feelings (TOTs) to the target-retrievability hypothesis. Familiarity of the cues was contrasted to memorability of the targets in a paired-associate design (e.g., A-B A-B, A-B A-B', A-B A-D, A-B C-D), in which the number of repetitions of the cue A terms was dissociated from the memorability of the target B terms. Little support was found for the target-retrievability hypothesis, because in none of the 4 experiments were FKJs related to target memorability. In one experiment, an omnibus retrieval hypothesis (which implicates total retrieval rather than just correct retrieval) and the cue-familiarity hypothesis produced isomorphic predictions that were borne out by the FKJ and TOT results. All 4 experiments supported the cue-familiarity hypothesis, because FKJs and TOTs were directly related to the number of presentations (and thereby the familiarity) of the cues.     

Strength,toughness and R-curve behaviour of SiC whisker-reinforced composite Si3N4 with reference to monolithic Si3N4

The flexural strength and fracture toughness of 30 vol% SiC whisker-reinforced Si₃N₄ material were determined as a function of temperature from 25 to 1400°C in an air environment. It was found that both strength and toughness of the composite material were almost the same as those of the monolithic counterpart. The room-temperature strength was retained up to 1100°C; however, appreciable strength degradation started at 1200°C and reached a maximum at 1400°C due to stable crack growth. In contrast, the fracture toughness of the two materials was independent of temperature with an average value of 5.66 MPam^1/2. It was also observed that the composite material exhibited no rising R-curve behaviour at room temperature, as was the case for the monolithic material. These results indicate that SiC whisker addition to the Si₃N₄ matrix did not provide any favourable effects on strength, toughness and R-curve behaviour.