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A quasi-linear constitutive relation for arterial wall materials.

In this work, considering that under physiological conditions the arteries are subjected to a large initial static deformation and additional small dynamical displacements are superimposed on this initial field, a Boltzman type of constitutive relation is presented for arterial wall materials. For its simplicity in the analysis, the material under investigation is assumed to be isotropic, incompressible and constitutionally nonlinear in Green deformation tensor but linear in the history of deformation tensor. The numerical values of elastic coefficients are obtained by comparing the analytical results with experiments by Simon et al. [Circ. Res. 30, 491-500 (1972)]. For the determination of viscoelastic coefficients we studied the propagation of torsional waves in a prestressed thin cylindrical tube and proposed a method of determining the viscoelastic coefficients.

Arteries↗

Effect of maturity and processing on total, insoluble and soluble dietary fiber contents of Indian green leafy vegetables.

Vegetables are very important in the diet and provide minerals, vitamins, antioxidants and fiber. A beneficial role of dietary fiber in human nutrition is known but the data on the effect of maturity and processing of Indian vegetables on dietary fiber and its fractions are not available. Hence, the present study was undertaken to generate data on total (TDF), insoluble (IDF) and soluble (SDF) dietary fiber contents of green leafy vegetables (GLV) and to assess the effect of leaf maturity and cooking on these parameters. Sixteen GLV (namely, agathi, alternanthera, amaranth, basella, cabbage, colocasia, coriander, curry leaves, drumstick, fenugreek, hibiscus, mint, portulaca, rumex, spinach and tender tamarind leaves) were analysed for TDF, IDF and SDF by the enzymatic and gravimetric method of the Association of Official Analytical Chemists. Among the GLV analysed, the TDF and IDF contents were the lowest (2.5 g% and 1.6 g%) in basella and were the highest (16.3 g% and 13.4 g%) in curry leaves. The SDF content ranged from 0.7 g% in spinach to 2.9 g% in curry leaves. The SDF as a percentage of the TDF ranged from 11.3% in tender tamarind leaves to 36.0% in basella, but the majority of GLV had around 25% of the TDF as SDF. Significant (P < 0.01) variation was observed between inter-species and intra-species in TDF, IDF and SDF contents of all the GLV. As the leaf matured from tender to mature and to coarse stage, the TDF and IDF contents of amaranth, basella, hibiscus, rumex and spinach increased significantly (P < 0.05), The SDF content significantly (P < 0.05) increased from tender to mature stage in all five GLV, but there was no further increase from mature to coarse stage except in rumex, where a significant (P < 0.01) increase (25%) was observed. Processing/cooking of GLV had no significant effect on their TDF, IDF and SDF contents.

Dietary Fiber↗

Resolution of viable and membrane-compromised bacteria in freshwater and marine waters based on analytical flow cytometry and nucleic acid double staining.

The membrane integrity of a cell is a well-accepted criterion for characterizing viable (active or inactive) cells and distinguishing them from damaged and membrane-compromised cells. This information is of major importance in studies of the function of microbial assemblages in natural environments, in order to assign bulk activities measured by various methods to the very active cells that are effectively responsible for the observations. To achieve this task for bacteria in freshwater and marine waters, we propose a nucleic acid double-staining assay based on analytical flow cytometry, which allows us to distinguish viable from damaged and membrane-compromised bacteria and to sort out noise and detritus. This method is derived from the work of S. Barbesti et al. (Cytometry 40:214-218, 2000) which was conducted on cultured bacteria. The principle of this approach is to use simultaneously a permeant (SYBR Green; Molecular Probes) and an impermeant (propidium iodide) probe and to take advantage of the energy transfer which occurs between them when both probes are staining nucleic acids. A full quenching of the permeant probe fluorescence by the impermeant probe will point to cells with a compromised membrane, a partial quenching will indicate cells with a slightly damaged membrane, and a lack of quenching will characterize intact membrane cells identified as viable. In the present study, this approach has been adapted to bacteria in freshwater and marine waters of the Mediterranean region. It is fast and easy to use and shows that a large fraction of bacteria with low DNA content can be composed of viable cells. Admittedly, limitations stem from the unknown behavior of unidentified species present in natural environments which may depart from the established permeability properties with respect to the fluorescing dyes.

Bacteria↗

Differential rates of gene expression monitored by green fluorescent protein.

The use of green fluorescent protein (GFP) as a reporter gene has made a broad impact in several areas, especially in studies of protein trafficking, localization, and expression analysis. GFP's many advantages are that it is small, autocatalytic, and does not require fixation, cell disruption, or the addition of cofactors or substrates. Two characteristics of GFP, extreme stability and chromophore cyclization lag time, pose a hindrance to the application of GFP as a real-time gene expression reporter in bioprocess applications. In this report, we present analytical methods that overcome these problems and enable the temporal visualization of discrete gene regulatory events. The approach we present measures the rate of change in GFP fluorescence, which in turn reflects the rate of gene expression. We conducted fermentation and microplate experiments using a protein synthesis inhibitor to illustrate the feasibility of this system. Additional experiments using the classic gene regulation of the araBAD operon show the utility of GFP as a near real-time indicator of gene regulation. With repetitive induction and repression of the arabinose promoter, the differential rate of GFP fluorescence emission shows corresponding cyclical changes during the culture.

Arabinose↗

Effect of the roasting method on the content of 5-hydroxytryptamides of carboxylic acids in roasted coffee beans.

Coffee beans of Coffea liberica (robusta) variety were roasted using convection and microwave heating. For roasting we used green coffee beans of 7.5% moisture content, and beans wetted to 10% moisture content and dried to 5% moisture content. The content of 5-hydroxytryptamides of carboxylic acids C-5-HT (determined by TLC) as the index of substances irritating alimentary canal was investigated in the roasted beans, depending on the bean treatment before roasting and applied roasting method. Analytical results show that predrying of the coffee beans caused 15-30% loss of C-5-HT, depending on the applied drying conditions. The content of C-5-HT in the roasted beans depended on the roasting method and preliminary treatment of the beans prior to roasting. A higher C-5-HT loss occurred in the case of beans subjected to two-stage processing, predrying and roasting. Convection roasting caused higher degradation of C-5-HT than microwave roasting.

Carboxylic Acids↗

Analyses of linker histone--chromatin interactions in situ.

Recent studies, using cytometric techniques based on fluorescence microscopy, have provided new information on how linker histones interact with chromatin in vivo or in situ. In particular, the use of green fluorescent proteins (GFPs) has enabled detailed studies of how individual H1 subtypes, and specific motifs in them, interact with chromatin in vivo. Furthermore, the development of cytochemical methods to study the interaction between linker histones and chromatin using DNA-binding fluorochromes as indirect probes for linker histone affinity in situ, in combination with highly sensitive and specific analytical methods, has provided additional information on the interactions between linker histones and chromatin in several cell systems. Such results verified that linker histones have a substantially higher affinity for chromatin in mature chicken erythrocytes than in frog erythrocytes, and they also indicated that the affinity decreased during differentiation of the frog erythrocytes. Furthermore, in cultured human fibroblasts, the linker histones showed a relatively high affinity for chromatin in interphase, whereas it showed a significantly lower affinity in highly condensed metaphase chromosomes. This method also enables the analysis of linker histone affinity for chromatin in H1-depleted fibroblasts reconstituted with purified linker histones. No consistent correlation between linker histone affinity and chromatin condensation has so far been detected.

Cell Nucleus↗

Microfluidic system for dielectrophoretic separation based on a trapezoidal electrode array.

This paper presents a novel microfluidic device for dielectrophoretic separation based on a trapezoidal electrode array (TEA). In this method, particles with different dielectric properties are separated by the device composed of the TEA for the dielectrophoretic deflection of particles under negative dielectrophoresis (DEP) and poly(dimethylsiloxane)(PDMS) microfluidic channel with a sinuous and expanded region. Polystyrene microparticles are exposed to an electric field generated from the TEA in the microfluidic channel and are dielectrophoretically focused to make all of them line up to one sidewall. When these particles arrive at the region of another TEA for dielectrophoretic separation, they are separated having different positions along the perpendicular direction to the fluid flow due to their different dielectrophoretic velocities. To evaluate the separation process and performance, both the effect of the flow rate on dielectrophoretic focusing and the influence of the number of trapezoidal electrodes on dielectrophoretic separation are investigated. Now that this method utilizes the TEA as a source of negative DEP, non-specific particle adhering to the electrode surface can be prevented; conventional separation approaches depending on the positive DEP force suffer from this problem. In addition, since various particle types are continuously separated, this method can be easily applicable to the separation and analysis of various dielectric particles with high particle recovery and selectivity.

Dimethylpolysiloxanes↗

Investigation into the effects of temperature and stirring rate on the solid-phase extraction of diuron from water using a C18 extraction disk.

A novel experimental method for determining the equilibrium constant, Keq, and the uptake rate constant, kup, for the solid-phase extraction (SPE) of diuron from water using a C18 Empore extraction disk is reported. Log Keq and log kup are determined at 7.0, 11.0, 18.0 and 23.0 degrees C and for stirring rates of 100, 200 and 400 rpm. From a Van 't Hoff plot of log Keq versus T-1 the enthalpy of sorption, delta H0, is shown to be negative which indicates that the thermodynamic process of uptake is exothermic. The rate of stirring has no effect on log Keq over the temperature range 7.0-23.0 degrees C. The enthalpy of activation, delta H0, calculated from Arrhenius plots of log kup versus T-1 at 100, 200 and 400 rpm show that the kinetic process of uptake is endothermic. At 100 rpm the rate of uptake is limited by the aqueous diffusion of diuron. At 200 rpm or greater the aqueous diffusion layer around the disk is sufficiently small to prevent diffusion from being a limiting factor. The method described in this paper is limited to the analysis of analytes that contain a significant UV chromophore and are relatively soluble in water, but it can also be used to investigate pH and salinity effects on the SPE of diuron from water.

Chromatography, Liquid↗

Comparison of yeast and mold counts by spiral, pour, and streak plate methods.

Foods with naturally occurring yeast and mold were used in a comparative study of 4 plating techniques. Oats, green beans, and cheese were incubated for 3 and 5 days at 25 degrees C. Although 3 days of incubation would be sufficient for yeast, 5 days were necessary for the mold count. The percent recovery of yeast and molds by the spiral and streak methods ranged from 200 to 357% compared with the pour plating method described in the Bacteriological Analytical manual (BAM). The spiral method had the highest overall recovery and the lowest replicate plating error.

Cheese↗

Improved gas chromatographic analysis of organophosphorus pesticides with pulsed splitless injection.

Gas chromatographic (GC) analysis of 6 organophosphorus pesticides (methamidophos, acephate, omethoate, diazinon, dimethoate, and chlorpyrifos) was performed with cool on-column, splitless, and pulsed splitless injections and with nitrogen-phosphorus or mass-selective detection. The pulsed splitless technique uses a high column flow rate during injection to sweep the sample out of the inlet rapidly, reducing analyte loss due to adsorption or thermal decomposition. After injection, the column flow rate is automatically reduced to normal values for chromatographic analysis. Pesticide recoveries for splitless and pulsed splitless injections were determined by comparison of GC peak areas with those obtained with cool on-column injection. With conventional splitless injection at a column flow rate of 5 mL/min, recoveries of acephate, omethoate, and methamidophos were only 57, 63, and 71%, respectively. Pulsed splitless methods with very fast injection flow rates dramatically improved recoveries, with all 6 pesticides falling in the 97-102% range. Because column flow rates are much less for GC with mass spectral detection (GC/MS), recoveries with splitless injection were lower and improvements with pulsed splitless injection were less dramatic for GC/MS. When splitless injection was used, recoveries of the 6 pesticides spiked into a green bean matrix were better than those of pesticides dissolved in pure solvent, presumably because matrix compounds compete with pesticides for active sites in the inlet. By using pulsed splitless injection of solvent standards with very fast initial column flow rates, systematic analyte losses in the inlet were eliminated, making recoveries of pesticides from solvent and green bean matrix very similar.

Chromatography, Gas↗

Metal dispersion and transportational activities using food crops as biomonitors.

The multielement (Al, Ca, Cd, Ce, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Si, and Zn) levels of various common vegetables (bean, broccoli, cabbage, cauliflower, lettuce, marrow, onion, parsnip, spinach, sprouts, sweet corn, and tomato); fruits (grape and strawberry); herbs (garlic, lemon balm, marjoram, mint, rosemary and tarragon); local pasture species and surface soils collected from a commercial garden centre located within a distance of 30 m of the London Orbital Motorway (M25) is presented. Comparative values are given from a background area, namely a domestic garden located in the North Yorkshire Dales National Park area. Analysis was undertaken by inductively coupled plasma optical emission spectrometry (ICP-OES) and inductively coupled plasma-source mass spectrometry (ICP-MS) with quality control assessment using four international biological reference materials; BCR:CRM 62 Olive Leaves, NIST 1575 Pine Needles, NIST 1573 Tomato Leaves, and NIST 1572 Citrus Leaves. Inter-analytical method comparison is given using two methods of ICP-MS; namely conventional pneumatic nebulisation of sample solution, and direct solids analysis by laser ablation; and neutron activation analysis methods (NAA). For the elements listed there is a good precision obtained by ICP-MS and NAA. In particular levels of < +/- 1-10% (rsd) are obtained. Comparison of data with certified values and other analytical methods are generally of very good agreement. Lead levels in background areas ranged from 0.0008 to 0.340 microgram/g (fresh weight) for plant material; with the lead magnitude greater for grasses > herbs > vegetables > cereals > fruits. Measured values are in good agreement with reported literature values. The lowest Pb values are for marrow, lettuce, tomato and sweet corn samples (approximately 0.001-0.021 microgram/g). 'Green' leaf material levels were approximately 0.02-0.10 microgram/g (i.e. sprouts and cabbage). Root vegetables contain higher levels, approximately 0.02-0.125 microgram/g (especially carrot), reflecting possible metal uptake from soil. The highest vegetable Pb values are for leek and onion (approximately 0.35 microgram/g). Background values are also provided for nineteen elements (Al, As, B, Ba, Br, Cd, Co, Cr, Cu, Fe, Li, Mn, Mo, Ni, Rb, Se, Sr, V, and Zn). Exposure to motor vehicle activities at a site some 30 m from the M25 shows only significant increases in Pb for unwashed plant material and surface soils. Typically Pb levels of 40-80% can be removed by washing plant surfaces resulting in metal levels similar to background areas.(ABSTRACT TRUNCATED AT 400 WORDS)

Air Pollutants↗

The inverse problem of a Gaussian convolution and its application to the finite size of the measurement chambers/detectors in photon and proton dosimetry.

A Gaussian convolution kernel K is deduced as a Green's function of a Lie operator series. The deconvolution of a Gaussian kernel is developed by the inverse Green's function K(-1). A practical application is the deconvolution of measured profiles Dm(x) of photons and protons with finite detector size to determine the profiles Dp(x) of point-detectors or Monte Carlo Bragg curves of protons. The presented algorithms work if Dm(x) is either an analytical function or only given in a numerical form. Some approximation methods of the deconvolution are compared (differential operator expansion to analytical adaptations of 2 x 2 cm2 and 4 x 4 cm2 profiles, Hermite expansions to measured 6 x 6 cm2 and 20 x 20 cm2 profiles and Bragg curves of 80/180 MeV protons, FFT to an analytical 4 x 4 cm2 profile). The inverse problem may imply ill-posed problems, and, in particular, the use of FFT may be susceptible to them.

Algorithms↗

Determination of sulfadoxine concentrations in whole blood using C18 solid-phase extraction, sodium dodecyl sulfate and dimethylaminocinnamaldehyde.

A simple method is described for the extraction and subsequent analysis of sulfadoxine in human whole blood using a solid-phase extraction technique and colorimetric reaction. This procedure utilizes the micellar properties of sodium dodecyl sulfate to: (1) extract sulfadoxine from a C18 solid-phase sample-preparation column; (2) enhance the colorimetric reaction produced by the addition of p-dimethylaminocinnamaldehyde (DMAC); and (3) provide stability to the coloured product generated by the reaction of sulfadoxine with DMAC. The intense, violet-red colour reaction can be conveniently used for qualitative and semiquantitative visual interpretations of sulfadoxine levels. Under the assay conditions, drug concentrations in the blood of subjects receiving sulfadoxine were determined from absorbance measurements. These results correlated well with the sulfadoxine levels determined from high-performance liquid chromatographic analysis. Important advantages of the procedure include the ability to evaluate small samples of whole blood (100 microliters), the minimal use of organic solvents, no sophisticated instrumentation, and formation of a stable, coloured reaction product. The method proved to be a suitable field assay for determining whole-blood levels of sulfonamides in the concentration range from 5 to 100 micrograms ml-1.

Chemistry Techniques, Analytical↗

Improved method to track chlorophyll degradation.

An analytical method capable of identifying >30 chlorophyll-related compounds in plant extracts has been developed. The method employs liquid chromatography coupled to UV-vis, MS, and MS/MS detection. It can be applied without modification to analyze natural chlorophyll degradation products and other metalloporphyrines. It was successfully applied to identify chlorophyll derivatives found in rehydrated spinach powder and conventionally canned and Veri-Green-processed beans. In the Veri-Green-processed beans several degradation products were identified that are zinc-containing analogues to the chlorophyll derivatives found in vegetables after conventional canning. They have been characterized by liquid chromatography and mass spectrometry.

Chlorophyll↗

Polarization state of the optical near field.

The polarization state of the optical electromagnetic field lying several nanometers above complex dielectric-air interfaces reveals the intricate light-matter interaction that occurs in the near-field zone. From the experimental point of view, access to this information is not direct and can only be extracted from an analysis of the polarization state of the detected light. These polarization states can be calculated by different numerical methods, well suited to near-field optics. In this paper, we apply two different techniques (localized Green's function method and differential theory of gratings) to separate each polarization component associated with both electric and magnetic optical near fields produced by nanometer sized objects. A simple dipolar model is used to get an insight into the physical origin of the near-field polarization state. In a second stage, accurate numerical simulations of field maps complete data produced by analytical models. We conclude this study by demonstrating the role played by the near-field polarization in the formation of the local density of states.

Journal Article↗

Analysis of pesticide residues in mixed fruit and vegetable extracts by direct sample introduction/gas chromatography/tandem mass spectrometry.

Direct sample introduction (DSI), or "dirty sample injection," was investigated in the determination of 22 diverse pesticide residues in mixed apple, green bean, and carrot extracts by benchtop gas chromatography/tandem mass spectrometry (DSI/GC/MS-MS). The targeted pesticides, some of which were incurred in the samples, included chlorpyrifos, azinphos-methyl, parathion-methyl, diazinon, terbufos, p,p'-DDE, endosulfan sulfate, carbofuran, carbaryl, propargite, bifenthrin, dacthal, trifluralin, metalaxyl, pendimethalin, atrazine, piperonyl butoxide, diphenylamine, vinclozolin, chlorothalonil, quintozene, and tetrahydrophthalimide (the breakdown product of captan). The analytical DSI method entailed the following steps: (1) blend 30 g sample with 60 mL acetonitrile for 1 min in a centrifuge bottle; (2) add 6 g NaCl and blend 30 s; (3) centrifuge for 1-2 min; (4) add 5 mL upper layer to 1 g anhydrous MgSO4 in a vial; and (5) analyze 11 microL extract, using DSI/GC/MS-MS. Sample cleanup is not needed because GCIMS-MS is exceptionally selective for the targeted analytes, and nonvolatile coextracted matrix components do not contaminate the injector or the GC/MS-MS system. Average recoveries of the pesticides were 103 +/- 7% with relative standard deviations of 14 +/- 5% on average, and limits of detection were <2 ng/g for nearly all pesticides studied. The DSI/GC/ MS-MS approach for targeted pesticides is quantitative, confirmatory, sensitive, selective, rugged, rapid, simple, and inexpensive.

Calibration↗

Electron-impact and glow-discharge ionization LC-MS analysis of green tea tincture.

A liquid chromatography-particle-beam mass spectrometer (LC-PB/MS) with interchangeable electron-impact (EI) and glow-discharge (GD) ion sources was evaluated for future application in analysis of botanical extracts. In this work a green tea tincture was characterized for a series of catechin components (catechin, epicatechin, epigallocatechin, and epigallocatechin gallate (EGCG)) and caffeine. Special emphasis was given to EGCG and caffeine, because they are important in determining the possible health effects of the green tea. The effects of instrument operating conditions were evaluated for the EI and GD ionization sources to determine their effect on analyte intensities and fragmentation patterns. These studies furnished information about the effects of these conditions in determining possible ionization pathways in the two ion sources. The mass spectra of these compounds obtained with the GD ion source are EI-like in appearance, with clearly identified molecular ions and fragmentation patterns that are easily rationalized. The absolute limits of detection for EGCG and caffeine were, respectively, 11 ng and 0.77 ng for the EI source and 3.2 ng and 0.61 ng for the GD source. The PB/EIMS and PB/GDMS combinations can be operated in a flow-injection mode, wherein the analyte is injected directly into the mobile phase, or coupled to high-performance liquid chromatography (HPLC), enabling LC-MS analysis of complex mixtures. A reversed-phase chromatographic separation of the green tea tincture was performed on a commercial C18 column using a gradient of water (containing 0.1% TFA) and ACN. Quantification of EGCG and caffeine was performed by the standard addition method. The amounts of EGCG and caffeine in the tested green tea tincture were each approximately 14 mg mL-1.

Caffeine↗

Comparison among differential pulse voltammetry, amperometric biosensor, and HPLC/DAD analysis for polyphenol determination.

Polyphenols are widespread in vegetables and fruits. They can play an important role in human diet and health, and they influence the sensorial properties of many foods, and act as natural antioxidants. This study was conducted using HPLC/DAD, tyrosinase biosensor, and differential pulse voltammetry (DPV) analyses to detect polyphenolic compounds in natural complex matrices. The analyses were applied to a series of both standards and natural extracts derived from grape, olives, and green tea. The pure compounds include phenolic acids, flavones, flavonols, catechins, tannins, and oleuropein. HPLC/DAD, DPV, and the biosensor approach were used as independent analytical techniques. Bare graphite screen-printed electrodes were employed in DPV and in the biosensor analysis. The most accurate data were obtained by HPLC/DAD analysis, while the DPV approach using screen-printed electrodes could represent a quick screening method for the determination of polyphenols in natural extracts. Use of the biosensor for the analysis of complex matrices needs further study in order to improve its performance.

Biosensing Techniques↗