Cell detection and counting through cell lysate impedance spectroscopy in microfluidic devices

Cell-based microfluidic devices have attracted interest for a wide range of applications. While optical cell counting and flow cytometry-type devices have been reported extensively, sensitive and efficient non-optical methods to detect and quantify cells attached over large surface areas within microdevices are generally lacking. We describe an electrical method for counting cells based on the measurement of changes in conductivity of the surrounding medium due to ions released from surface-immobilized cells within a microfluidic channel. Immobilized cells are lysed using a low conductivity, hypotonic media and the resulting change in impedance is measured using surface patterned electrodes to detect and quantify the number of cells. We found that the bulk solution conductance increases linearly with the number of isolated cells contributing to solution ion concentration. The method of cell lysate impedance spectroscopy is sensitive enough to detect 20 cells microL(-1), and offers a simple and efficient method for detecting and enumerating cells within microfluidic devices for many applications including measurement of CD4 cell counts in HIV patients in resource-limited settings. To our knowledge, this is the most sensitive approach using non-optical setups to enumerate immobilized cells. The microfluidic device, capable of isolating specific cell types from a complex bio-fluidic and quantifying cell number, can serve as a single use cartridge for a hand-held instrument to provide simple, fast and affordable cell counting in point-of-care settings.     

Monitoring conformational changes in protein complexes using chemical cross-linking and Fourier transform ion cyclotron resonance mass spectrometry: the effect of calcium binding on the calmodulin-melittin complex

Calmodulin is an EF hand calcium binding protein. Its binding affinities to various protein/peptide targets often depend on the conformational changes induced by the binding of calcium. One such target is melittin, which binds tightly to calmodulin in the presence of calcium, and inhibits its function. Chemical cross-linking combined with Fourier transform ion cyclotron resonance mass spectrometry has been employed to investigate the coordination of calmodulin and melittin in the complex at different concentrations of calcium. This methodology can be used to monitor structural changes of proteins induced by ligand binding, and study the effects these changes have on non- covalent interactions between proteins. Cross-linking results indicate that the binding place of the first melittin in the calcium free calmodulin form is the same as in the calcium loaded calmodulin/melittin complex.     

Sage,Salvia officinalis L., Constituents,Hepatoprotective Activity,and Cytotoxicity Evaluations of the Essential Oils Obtained from Fresh and Differently Timed Dried Herbs: A Comparative Analysis

Sage, Salvia officinalis L., is used worldwide as an aromatic herb for culinary purposes as well as a traditional medicinal agent for various ailments. Current investigations exhibited the effects of extended dryings of the herb on the yields, composition, oil quality, and hepatoprotective as well as anti-cancer biological activities of the hydrodistillation-obtained essential oils from the aerial parts of the plant. The essential oils’ yields, compositions, and biological activities levels of the fresh and differently timed and room-temperature dried herbs differed significantly. The lowest yields of the essential oil were obtained from the fresh herbs (FH, 631 mg, 0.16%), while the highest yield was obtained from the two-week dried herbs (2WDH, 1102 mg, 0.28%). A notable decrease in monoterpenes, with increment in the sesquiterpene constituents, was observed for the FH-based essential oil as compared to all the other batches of the essential oils obtained from the different-timed dried herbs. Additionally, characteristic chemotypic constituents of sage, i.e., α-pinene, camphene, β-pinene, myrcene, 1, 8-cineole, α-thujone, and camphor, were present in significantly higher proportions in all the dried herbs’ essential oils as compared to the FH-based essential oil. The in vivo hepatoprotective activity demonstrated significant reductions in the levels of AST, ALT, and ALP, as well as a significant increase in the total protein (p < 0.05) contents level, as compared to the acetaminophen (AAP) administered experimental group of rats. A significant reduction (p < 0.05) in the ALT level was demonstrated by the 4WDH-based essential oil in comparison to the FH-based essential oil. The levels of creatinine, cholesterol, and triglycerides were reduced (p < 0.05) in the pre-treated rats by the essential oil batches, with non-significant differences found among them as a result of the herbs dryings based oils. A notable increase in the viability of the cells, and total antioxidant capacity (TAOxC) levels, together with the reduction in malondialdehyde (MDA) levels were observed by the essential oils obtained from all the batches as compared with the AAP-treated cell-lines, HepG-2, HeLa, and MCF-7, that indicated the in vitro hepatoprotective effects of the sage essential oils. However, significant improvements in the in vivo and in vitro hepatoprotective activities with the 4WDH-based oil, as compared to all other essential oil-batches and silymarin standard demonstrated the beneficial effects of the drying protocol for the herb for its medicinal purposes.     

Gauge Invariance of a Critical Number of Flavours in QED3

The fermion propagator in an arbitrary covariant gauge can be obtained from the Landau gauge result via a Landau–Khalatnikov–Fradkin transformation. This transformation can be written in a practically useful form in both configuration and momentum space. It is therefore possible to anticipate effects of a gauge transformation on the propagator’s analytic properties. These facts enable one to establish that if a critical number of flavours for chiral symmetry restoration and deconfinement exists in noncompact QED3, then its value is independent of the gauge parameter. This is explicated using simple forms for the fermion–photon vertex and the photon vacuum polarisation. The illustration highlights pitfalls that must be avoided in order to arrive at valid conclusions. Landau gauge is seen to be the covariant gauge in which the propagator avoids modification by a non-dynamical gauge-dependent exponential factor, whose presence can obscure truly observable features of the theory.     

Towards a classification of energy aware MAC protocols for wireless sensor networks

Power management is an important issue in wireless sensor networks (WSNs) because wireless sensor nodes are usually battery powered, and an efficient use of the available battery power becomes an important concern specially for those applications where the system is expected to operate for long durations. This necessity for energy efficient operation of a WSN has prompted the development of new protocols in all layers of the communication stack. Provided that, the radio transceiver is the most power consuming component of a typical sensor node, large gains can be achieved at the link layer where the medium access control (MAC) protocol controls the usage of the radio transceiver unit. MAC protocols for sensor networks differ greatly from typical wireless networks access protocols in many issues. MAC protocols for sensor networks must have built‐in power conservation, mobility management, and failure recovery strategies. Furthermore, sensor MAC protocols should make performance trade‐off between latency and throughput for a reduction in energy consumption to maximize the lifetime of the network. This is in general achieved through duty cycling the radio transceiver. Many MAC protocols with different objectives were proposed for wireless sensor networks in the literature. Most of these protocols take into account the energy efficiency as a main objective. There is much more innovative work should be done at the MAC layer to address the hard unsolved problems. In this paper, we first outline and discuss the specific requirements and design trade‐offs of a typical wireless sensor MAC protocol by describing the properties of WSN that affect the design of MAC layer protocols. Then, a typical collection of wireless sensor MAC protocols presented in the literature are surveyed, classified, and described emphasizing their advantages and disadvantages whenever possible. Finally, we present research directions and identify open issues for future medium access research. Copyright © 2009 John Wiley & Sons, Ltd.     

Interactions between cationic gemini/conventional surfactants with polyvinylpyrrolidone: Specific conductivity and dynamic light scattering studies

The interaction between bis(hexadecyldimethylammonium)hexane dibromide (16-6-16), bis(tetradecyldimethylammonium)hexane dibromide (14-6-14), their conventional counterparts cetyltrimethylammonium bromide (CTAB) and tetradecyltrimethylammonium bromide (TTAB) with polyvinylpyrrolidone (PVP) was investigated using the conductivity technique. The results show that gemini surfactants interact strongly with PVP as compared to conventional surfactants. The results also reveal that the surfactants with shorter hydrocarbon chain interact weakly as those of longer hydrocarbon chain. The interactions of 16-6-16 and 14-6-14 and their conventional counterparts with PVP were also studied using dynamic light scattering (DLS) measurements. We have also highlighted the effect of surfactant–polymer interactions on the dispersion force in the solution. Critical aggregation concentration (cac) and critical micelle concentration (cmc) were obtained using the conductivity data. The degrees of micelle ionization and free energies associated with aggregation, micellization, and transfer have also been evaluated and discussed.     

Green synthesis and characterization of polymer-stabilized silver nanoparticles

Silver nanoparticles (Ag-NPs) were synthesized using a facile green chemistry synthetic route. The reaction occurred at ambient temperature with four reducing agents introduced to obtain nanoscale Ag-NPs. The variables of the green synthetic route, such as acidity, concentration of starting materials, and molar ratio of reactants were optimized. Dispersing agents were employed to prevent Ag-NPs from aggregating. Advanced instrumentation techniques, such as X-ray powder diffraction (XRD), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), ultraviolet–visible spectroscopy (UV–vis), and phase analysis light scattering technique (ZetaPALS) were applied to characterize the morphology, particle size distribution, elemental composition, and electrokinetic behavior of the Ag-NPs. UV–vis spectra detected the characteristic plasmon at approximately 395–410 nm; and XRD results were indicative of face-centered cubic phase structure of Ag. These particles were found to be monodispersed and highly crystalline, displaying near-spherical appearance, with average particle size of 10.2 nm using citrate or 13.7 nm using ascorbic acid as reductants from particle size analysis by ZetaPALS, respectively. The rapid electrokinetic behavior of the Ag was evaluated using zetapotential (from −40 to −42 mV), which was highly dependant on nanoparticle acidity and particle size. The current research opens a new avenue for the green fabrication of nanomaterials (including variables optimization and aggregation prevention), and functionalization in the field of nanocatalysis, disinfection, and electronics.     

Spectrophotometric Determination of Piroxicam via Chelation with Fe(III) in Commercial Dosage Forms

The objective of this work is to develop and validate spectrophotometric method for the determination of piroxicam in commercial dosage forms. The method is based on the chelation of the drug with Fe(III) to form pink coloured metal chelate at room temperature which absorbs maximally at 504 nm. Beer's law is obeyed over the concentration range of 8–160 μg mL^?1 (A = 1.07 × 10^?3 + 7.75 × 10^?3 C). Under the optimized experimental conditions, proposed method is validated as per the International Conference on Harmonisation guidelines. The limits of detection and quantitation for the proposed method are 0.775 and 2.348 μg mL^?1, respectively. The proposed method has been successfully applied to the determination of piroxicam in commercial dosage forms. The results are compared with the reference El‐Ries et al. spectrophotometric method.     

Preconcentration of Rare Earth Elements in Seawater with Chelating Resin Having Fluorinated β‐Diketone Immobilized on Styrene Divinyl Benzene for their Determination by ICP‐OES

An analytical method has been developed for the preconcentration of rare earth elements (REEs) in seawater for their determination by inductively coupled plasma optical emission spectrometry (ICP‐OES). An indigenously synthesized chelating resin was used for the preconcentration of (REEs) which was based on immobilization of fluorinated β‐diketone group on solid support styrene divinyl benzene. Sample solutions (adjusted to optimized pH) were passed through a polyethylene column packed with 250 mg of the resin. Experimental conditions consisting of pH, sample flow rate, sample volume and eluent concentration were optimized. The established method has been applied for the preconcentration of light, medium and heavy REEs in coastal sea water samples for their subsequent determination by (ICP‐OES). Percentage recoveries of La, Ce, Nd, Sm, Eu, Gd, Dy, Er, Yb and Lu were ≥ 95%, a preconcentration factor of 200 times, and relative standard deviations < 5% were achieved.