Potential use of chemical cues for colony-mate recognition in the big brown bat,Eptesicus fuscus

Bats should benefit from recognition of their roost-mates when colonies form stable social units that persist over time. We used Y-maze experiments and gas chromatography–olfactometry (GC-O) to evaluate whether female big brown bats Eptesicus fuscus (Chiroptera: Vespertilionidae) use chemical cues to distinguish among conspecifics. In dual-choice Y-maze experiments, females chose the scent of another female from their own roost over a conspecific female from a different roost in a majority of trials. Analysis of total body odors using GC-O suggests that individuals from a given colony may share a more common odor signature with roost-mates than with non-roost-mate conspecifics. Using four principle components derived from 15 odor variables, discriminant function analysis correctly assigned most individuals to the correct colony.     

Evaluation of the separate equilibrium processes that dictate the upper detection limit of neutral ionophore-based potentiometric sensors

The upper detection limit of polar ionophore-based ion-selective electrode membranes is predicted by utilizing the coextraction constant of dissociated electrolyte, the stability constant of the ionophore, and the membrane composition. The coextraction constant of dissociated electrolytes into the polar poly(vinyl chloride) membrane plasticized with o-nitrophenyl octyl ether (PVC-NPOE) is here measured by a novel approach. The sandwich membrane technique is utilized, with one membrane segment containing a lipophilic cation exchanger and the other containing an anion exchanger. This yields information about the coextraction constant and the free ion concentrations of the electrolyte in the two segments. Predictions correlate quantitatively with the upper detection limit observed for ion-selective electrodes based on the ionophores valinomycin, tert-butylcalix[4]arene tetraethyl ester, and calcimycin. The difficulties of the prediction of the upper detection limit for nonpolar poly(vinyl chloride) membranes plasticized with bis(2-ethylhexyl sebacate) (PVC-DOS) due to ion association are discussed in detail. A thermodynamic cycle experiment with a series of sandwich membranes shows that the principal processes governing the upper detection limit of PVC-DOS membranes are identical to those for the PVC-NPOE membranes. However, the stability of the ion pairs between the ionophore-metal ion complexes and the extracted anion are different from that of ion pairs formed between the same anion and the lipophilic anion exchanger. This makes it difficult to quantitatively predict the upper detection limit on the basis of simple apparent coextraction and complexation data alone. The approach reported herein is useful not only for mechanistic purposes but also to shed light onto the many cases where coextraction effects need to be understood but are not directly experimentally accessible.     

Testing the robustness of the job demands-resources model.

[Correction Notice: An erratum for this article was reported in Vol 14(2) of International Journal of Stress Management (see record 2007-07240-008). Several corrections of errata in a paper by the authors published in the 2006 issue of the International Journal of Stress Management are presented: (1) The sentence beginning on page 385, line 16 should read "To avoid identification problems, the error variance of Organizational commitment and Emotional load were constrained using the formula (1 - α) × sigma2." (2) The sentence on page 385, line 18, should be deleted. (3) Degrees of freedom for M3 in Table 2 were incorrectly reported. The correct degrees of freedom were 54. The corrected table is provided.] According to the job demands-resources (JD-R) model, job demands and resources evoke two relatively independent processes: health impairment and employee motivation. The robustness of the JD-R model was tested in two different occupational samples, the first of 654 Spanish employees and the second of 477 Dutch employees. Structural equation modeling analyses provided partial evidence for the two processes. Multigroup analyses showed that the structural paths of the model were invariant across countries, although the strength of the relationships differed. We conclude that the basic structure of the JD-R model is maintained, even when applied in different national and occupational contexts, when using different ways of gathering data (computerized versus paper and pencil), and when using slightly different measures to assess the key variables of the model. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Direct potentiometric information on total ionic concentrations

Polymeric membrane ion-selective electrodes exhibit an apparently super-Nernstian response at low sample activities if inner solutions are used that induce strong zero-current fluxes of primary ions toward the inner compartment. This is due to the limited ion fluxes in the aqueous boundary layer near the membrane. In the presence of labile complexes, the effective flux rate is increased and the emf depends on the total concentration of the ions. The concept is illustrated experimentally with calcium-selective electrodes based on the ionophore N,N-dicyclohexyl-N',N'-dioctadecyl-3-oxapentanediamide (ETH 5234) that either respond to total or free ion concentrations. Samples can be distinguished that contain varying levels of total calcium but are all buffered with EDTA to the same free calcium concentration of 5 x 10(-8) M.     

Sampling and quantifying invertebrates from drinking water distribution mains

Water utilities in the Netherlands aim at controlling the multiplication of (micro-) organisms by distributing biologically stable water through biologically stable materials. Disinfectant residuals are absent or very low. To be able to assess invertebrate abundance, methods for sampling and quantifying these animals from distribution mains were optimised and evaluated. The presented method for collecting invertebrates consists of unidirectionally flushing a mains section with a flow rate of 1 ms(-1) and filtering the flushed water in two separate flows with 500 microm and 100 microm mesh plankton gauze filters. Removal efficiency from mains was evaluated in nine experiments by collecting the invertebrates removed from the mains section by intensive cleaning immediately subsequent to sampling. Of 12 taxa distinguished, all except case-building Chironomidae larvae (2%) and Oligochaeta (30%) were removed well (51-75%). Retention of invertebrates in 100 microm filters was evaluated by filtering 39 filtrates using 30 microm filters. Except for flexible and small invertebrates such as Turbellaria (13%), Nematoda (11%) and Copepoda larvae (24%), most taxa were well retained in the 100 microm filters (53-100%). During sample processing, the method for taking sub-samples with a 10 ml pipette from the suspension of samples with high sediment concentrations was found to perform well in 75% of the samples. During a 2-year national survey in the Netherlands and consecutive investigations, the method appeared to be very suitable to assess the abundance of most invertebrate taxa in drinking water distribution systems and to be practicable for relatively inexperienced sampling and lab technicians. Although the numbers of small, less abundant or sessile taxa were not accurately assessed using the method, these taxa probably should not be the primary focus of monitoring by water utilities, as consumer complaints are not likely to be caused by these invertebrates. The accuracy of quantifying small invertebrates was further improved, however, by filtering the 100microm filtrate with a 30microm mesh plankton gauze filter.     

Fluorescence quenching-based assays for hydrolyzing enzymes. Application of time-resolved fluorometry in assays for caspase, helicase, and phosphatase

We have developed assay technologies to measure hydrolyzing enzymes based on homogeneous time-resolved fluorescence quenching (TruPoint). High sensitivity was obtained using fluorescent europium chelates as labels, internally quenched by suitable quenchers and released upon enzymatic reaction. This approach allows robust and sensitive monitoring of low enzyme activities. The assay technology and the choice of donor-acceptor pairs were evaluated in three different enzymatic assays, a protease related to apoptosis, helicase involved in DNA unwinding, and phosphatase having an important role in cellular signaling cascades. All the assays produced an increasing signal, were sensitive, and had a good dynamic range. There were significant differences in optimized quenchers for each of the assays depending on the size, flexibility, and rigidity of the substrates. Also, clear differences in the energy-transfer reactions, their requirements for spectral overlapping, ionic interactions, and energy-transfer distances were found. Each of the enzymatic assays was briefly tested in a high-throughput screening environment by analyzing signal dynamics and statistical relevance as Z' factors.     

Single-step formation of a biorecognition layer for assaying histidine-tagged proteins

The purpose of this work was to develop a simple procedure for the creation of a specific biorecognition layer for histidine-tagged (His-tagged) molecules. Such a layer was prepared by the spontaneous fusion of vesicles containing readily available plain (DOPC) and iminodiacetic acid (DOGS-NTA) phospholipids on a silica surface resulting in the formation of an NTA-containing supported lipid bilayer. The frequency surface acoustic waveguide device which supports Love waves was used to follow the real-time formation of the biorecognition layer. The mole percent of the DOGS-NTA phospholipids in the supported bilayer was optimized by following the kinetics of the fusion for the different NTA-containing lipids. Fluorescently labeled lipids were used with observations of the fluorescence recovery after photobleaching to confirm the presence of lipid bilayers. After saturating all NTA-molecules with Ni(2+), the binding of a His-tagged protein fragment within the concentration range of 0.04 and 0.4 mM to a 5 mol % DOGS-NTA/DOPC was detected; binding curves were used to calculate the apparent association constant k(on) = 2.56 x 10(4) M(-)(1) s(-)(1), dissociation constant k(off) = 1.3 x 10(-)(3) s(-)(1), and equilibrium constant k(eq) = 1.97 x 10(7) M(-)(1). The described method could find significant applications as a generic technique for preparing biorecognition layers for His-tagged proteins. In addition, the acoustic waveguide device, which provides high sensitivity together with flexibility in terms of the substrate material used, is shown to be an attractive alternative to direct optical biosensors.     

Dynamic diffusion model for tracing the real-time potential response of polymeric membrane ion-selective electrodes

A numerical solution for the prediction of the time-dependent potential response of a polymeric-based ion-selective electrode (ISE) is presented. The model addresses short- and middle-term potential drifts that are dependent on changes in concentration gradients in the aqueous sample and organic membrane phase. This work has important implications for the understanding of the real-time response behavior of potentiometric sensors with low detection limits and with nonclassical super-Nernstian response slopes. As a model system, the initial exposure of membranes containing the well-examined silver ionophore O,O' '-bis[2-(methylthio)ethyl]-tert-butylcalix[4]arene was monitored, and the large observed potential drifts were compared to theoretical predictions. The model is based on an approximate solution of the diffusion equation for both aqueous and organic diffusion layers using a numerical scheme (finite difference in time and finite elements in space). The model may be evaluated on the basis of experimentally available parameters and gives time-dependent information previously inaccessible with a simpler steady-state diffusion model. For the cases studied, the model gave a very good correlation with experimental data, albeit with lower than expected diffusion coefficients for the organic phase. This model may address numerous open questions regarding the response time and memory effects of low-detection-limit ion-selective electrodes and for other membrane electrodes where ion fluxes are relevant.     

Elimination of dimer formation in InIIIporphyrin-based anion-selective membranes by covalent attachment of the ionophore

The spontaneous hydroxy-bridged dimer formation of metalloporphyrins in ion-selective membranes gives rise to a short sensor lifetime (typically days), triggered by solubility problems, the occurrence of a super-Nernstian response slope, and a pH cross response. This dimer formation is eliminated here by covalent attachment of the ionophore to the polymer matrix. Specifically, two different indium(III)porphyrins containing polymerizable groups, the chloride-selective chloro(3-[18-(3-acryloyloxypropyl)-7,12-bis(1-methoxyethyl)-3,8,13,17-tetramethylporphyrin-2-yl]propyl ester)indium(III) and the nitrite-selective Chloro(5-(4-acryloyloxyphenyl)-10,15,20-triphenylporphyrinato)indium(III), were synthesized and copolymerized with methyl methacrylate and decyl methacrylate. The covalent attachment of the ionophore to the polymer matrix indeed prevents the metalloporphyrin from forming dimeric species, as confirmed by UV/visible spectroscopy. The ion-selective membranes with grafted indium porphyrin showed Nernstian response slopes to chloride, nitrite, perchlorate, and thiocyanate anions, with a selectivity comparable to membranes with freely dissolved or underivatized metalloporphyrin. The membranes containing grafted ionophores showed a lifetime of at least two months, apparently since crystallization of the poorly soluble dimeric species may no longer occur. This is one of the first examples where the covalent attachment of an ionophore drastically improves on a number of important sensor characteristics.