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Extraction Procedure (extraction + procedure)
Kinds of Extraction Procedure Selected AbstractsKINETICS OF SOYBEAN LIPOXYGENASES ARE RELATED TO pH, SUBSTRATE AVAILABILITY AND EXTRACTION PROCEDURESJOURNAL OF FOOD BIOCHEMISTRY, Issue 2 2008VERONICA S. CHEDEA ABSTRACT The kinetic patterns of pure soy lipoxygenase LOX-1 and crude or defatted soybean extracts containing LOX isoenzymes (LOX-1, LOX-2 and LOX-3) were studied by UV spectrometry at 234 and 280 nm, depending on their extraction and measurement conditions. Different pHs (from 6.0 to 9.0), corresponding to specific activation of LOX isoenzymes and the ratios of enzyme protein per substrate were used in order to evaluate the enzyme rates, as indicators of its affinity for substrate in different environments. The crude soy extract contained mainly LOX-1 activity (measured at 234 nm, at pH 9.0) and LOX-3, in an approximate ratio of 3:1. The LOX-2 activity was very low. The defatted extracts buffered at pH 6.8 and 7.1 showed a low LOX-1 and LOX 2 activity, but mostly LOX-3 activity (measured at 280 nm, at pH 7.1), with a mirror-type relation between the enzyme/substrate ratio and their enzymatic specific activity. The results suggest that defatting inhibits specifically the LOX-1 activity and indicate the possibility to modulate LOX activity by modifications of enzyme/substrate ratios and modifications of pH in the enzyme environment. PRACTICAL APPLICATIONS Because of the specific kinetic behaviors of the three different LOXs found in crude soy extracts involved in off-flavor generation, one can modulate the inhibition of these isoenzymes during soybean processing. Our experiments showed that pH variation could be a simple solution to inhibit the LOX isoenzymes, and therefore, the off-flavor generation. From the analytical point of view, the techniques described in this article are designed to be as simple as possible, and easy to use at large-scale level in food industry (food chain control). The idea is to minimize the number of separate chemical manipulations and, thereby, minimize errors. These studies can offer the background of further inhibition experiments in vitro using natural extracts. The LOX inhibition by natural antioxidants is related as well to pH and other factors influencing the enzyme's activity; this idea can be also valorized practically in the future. [source] Quantitative Analysis of Prometrine Herbicide by Liquid,Liquid Extraction Procedures Coupled to Electrochemical MeasurementsELECTROANALYSIS, Issue 6 2009V. Juarez Abstract A sensitive method is proposed for the preconcentration and quantification of the herbicide Prometrine (PROM) at a liquid-liquid interface employing square-wave voltammetry. The preconcentration stage was based on liquid-liquid extraction methodology and the PROM quantification was carried out from the peak current of square-wave voltammograms. Under the experimental conditions employed, linear calibration curves in the concentration range 1.0×10,6,M,5.0×10,5,M, with detection limit equal to 1.5×10,6,M were obtained without pretreatment of the samples. This linear range, as well as detection limit could be extended towards lower concentrations when a pretreatment procedure was employed. In this way, linearity of calibration curves between 8.0×10,8,M and 2.4×10,7,M and detection limit of 1.0×10,7,M, were observed. On the other hand, the standard addition method was also used as an alternative and an appropriated quantification technique for this system. A linear concentration range between 1.0×10,6,M and 2.7×10,5,M, with a correlation coefficient of 0.997, was obtained. This procedure has also a promising application in the separation of herbicides from other interferents, present in real samples, previous to their quantification. [source] Clustering revealed in high-resolution simulations and visualization of multi-resolution features in fluid,particle modelsCONCURRENCY AND COMPUTATION: PRACTICE & EXPERIENCE, Issue 2 2003Krzysztof Boryczko Abstract Simulating natural phenomena at greater accuracy results in an explosive growth of data. Large-scale simulations with particles currently involve ensembles consisting of between 106 and 109 particles, which cover 105,106 time steps. Thus, the data files produced in a single run can reach from tens of gigabytes to hundreds of terabytes. This data bank allows one to reconstruct the spatio-temporal evolution of both the particle system as a whole and each particle separately. Realistically, for one to look at a large data set at full resolution at all times is not possible and, in fact, not necessary. We have developed an agglomerative clustering technique, based on the concept of a mutual nearest neighbor (MNN). This procedure can be easily adapted for efficient visualization of extremely large data sets from simulations with particles at various resolution levels. We present the parallel algorithm for MNN clustering and its timings on the IBM SP and SGI/Origin 3800 multiprocessor systems for up to 16 million fluid particles. The high efficiency obtained is mainly due to the similarity in the algorithmic structure of MNN clustering and particle methods. We show various examples drawn from MNN applications in visualization and analysis of the order of a few hundred gigabytes of data from discrete particle simulations, using dissipative particle dynamics and fluid particle models. Because data clustering is the first step in this concept extraction procedure, we may employ this clustering procedure to many other fields such as data mining, earthquake events and stellar populations in nebula clusters. Copyright © 2003 John Wiley & Sons, Ltd. [source] Determination of Diazepam, Temazepam and Oxazepam at the Lead Film Electrode by Adsorptive Cathodic Stripping VoltammetryELECTROANALYSIS, Issue 17-18 2010Katarzyna Tyszczuk Abstract The determination of psychoactive 1,4-benzodiazepine drugs is of relevant interest in clinical, biomedical areas. Therefore a highly sensitive and simple voltammetric method for the determination of temazepam, diazepam and oxazepam at an in situ plated lead film electrode was developed. The method was successfully applied to the determination of diazepam and temazepam in pharmaceutical formulations with minimum sample manipulation and oxazepam in human urine samples without any separation steps. The determinations of oxazepam in human urine samples were performed in a flow system. Therefore a previous extraction procedure was not necessary to separate the active compound before its determination. [source] Simultaneous determination of six non-polar heterocyclic amines in meat samples by supercritical fluid extraction,capillary electrophoresis under fluorimetric detectionELECTROPHORESIS, Issue 13 2010Fernando De Andrés Abstract A novel, sensitive and selective method for the separation and quantification of a group of non-polar heterocyclic amines (9H-pyrido-[3,4-b] indole, norharmane; 1-methyl-9H-pyrido-[3,4-b] indole, harmane; 2-amino-9H-pyrido-[2,3-b] indole, A,C; 2-amino-3-methyl-9H-pyrido-[2,3-b] indole, MeA,C; 3-amino-1,4-dimethyl-5H-pyrido-[4,3-b] indole, Trp-P-1 and 3-amino-1-methyl-5H-pyrido-[4,3-b] indole, Trp-P-2) in commercial meat samples has been developed. This methodology is faster than others previously described. The method is based on the combination of a supercritical fluid extraction procedure, followed by the analysis of the extracted plug by CE with fluorescence detection. The supercritical fluid extraction procedure was optimized for the clean-up of the samples and the extraction of the analytes. For the electrophoretic separation, the effect of composition, pH and concentration of buffer, organic modifier content, pressure and time of injection, capillary temperature and voltage applied were studied. A 10,mmol/L formic acid,ammonium formate,ACN (10%, v/v) solution at pH 1.5 was selected as the running electrolyte. With 5-s hydrodynamic injection, linear responses in the range from 100 to 1000,ng/mL and detection limits ranging from 15.9 to 28.1,ng/mL were obtained for different amines in less than 13,min. ACN,water (1:1 in volume) was used as a sample solvent. Fluorescence detection enhances the sensitivity and avoids interferences coming from non-fluorescent compounds present in the matrices of the sample extracts. [source] Integrated microdevice for preconcentration and separation of a wide variety of compounds by electrochromatographyELECTROPHORESIS, Issue 3 2009Gaelle Proczek Abstract An integrated microdevice was developed to couple on-chip SPE to separation by channel electrochromatography. An acrylate-based monolith was synthesized within a glass microdevice by photoinitiated polymerization. It was used for both separation and preconcentration by direct injection on the head of the stationary phase or by confining the preconcentration step in a given zone of the stationary phase. The composition of the polymerization mixture was chosen to achieve a monolithic material containing both hydrophobic and charged moieties to ensure an electroosmotic flow for separation. As a consequence the extraction procedure occurs via hydrophobic and ionic interactions. Neutral, ionizable and charged compounds were successfully preconcentrated and separated within the microdevice through electrochromatographic mechanisms, highlighting the versatility of this device. The performance of the integrated microdevice was demonstrated with the preconcentration and separation of a mixture of PAHs for which a signal enhancement factor (SEF) of 270 was achieved within 120,s of preconcentration. In the case of charged and ionizable compounds, according to the electrolyte composition, contributions of both reverse-phase and ion-exchange mechanisms were used to perform effective electrochromatographic preconcentration. A SEF of 250 was obtained for the model-charged compound within 20,s of preconcentration. Finally, the potentials of on-chip preconcentrate and separate both neutral and ionized compounds have been demonstrated using a mixture of model compounds. [source] SPE and large-volume sample stacking in MEKC for determination of doxycycline in biological fluids: Comparison of direct injection to SPE-MEKCELECTROPHORESIS, Issue 21 2008Rade Injac Abstract A novel and simple method has been developed for the determination of doxycycline (DOX) in biological fluids. The method is based on SPE, large-volume sample stacking (LVSS) and MEKC with UV-DAD detection. Six SPE cartridges have been used in investigation for sample clean up and pre-concentration (Supelco® LC-8, LC-18, LC-SCX, and LC-WCX, as well as StrataÔ-X and X-C). DOX was determined on a 56,cm (effective length 50,cm)×50,,m id fused-silica capillary. The BGE was 20,mM borate buffer, pH 9.3, containing 80,mM SDS and 7.5%,v/v of methanol (30,s×50,mbar), and the temperature and voltage were 25°C and 30,kV, respectively. The analytical wavelength was set at 210,nm. Under optimized conditions it is possible to determine DOX in human serum, urine, semen, tears and saliva with recovery of 97.5% (RSD 2.5%). The method was shown to be sensitive (LOD is 1,,g/L) and precise (intra-day RSD 0.2 and 2.4%; inter-days 0.4 and 3.5% for migration time and peak area, respectively). Results for developed SPE-LVSS-MEKC were compared with LVSS-MEKC method with direct sample injection. The new LVSS-MEKC method is presented as a useful technique for rapid determination without extraction procedure of DOX in human urine and serum, using 80,mM of SDS, 10%,v/v of methanol and 40,mM borate buffer (pH 9.3; 30,s×50,mbar; 25°C; 30,kV; 350,nm), but not for the other biological fluids, according to lower sensitivity of the method and because of the sample composition. [source] Determination of flurbiprofen enantiomers in plasma using a single-isomer amino cyclodextrin derivative in nonaqueous capillary electrophoresis,ELECTROPHORESIS, Issue 17 2008Anne Rousseau Abstract A nonaqueous capillary electrophoresis (NACE) assay was developed for the separation and determination of flurbiprofen enantiomers in plasma samples using 6-monodeoxy-6-mono(3-hydroxy)propylamino-,-cyclodextrin as chiral selector. The nonaqueous background electrolyte was made up of 40,mM ammonium acetate in methanol (MeOH), and flufenamic acid was employed as internal standard. Solid-phase extraction was used for sample cleanup prior to the NACE separation. The NACE method reproducibility was optimized by evaluating different capillary washing sequences between runs. After having tested various conditions, trifluoroacetic acid (1,M) in MeOH was finally selected. Concerning the solid-phase extraction procedure, good and reproducible analyte recoveries were obtained using MeOH for protein denaturation and a polymeric phase combining hydrophobic interactions with anion exchange properties (Oasis® MAX) was selected as extraction sorbent. The method selectivity was not only demonstrated toward a blank plasma sample but also toward other non-steroidal anti-inflammatory drugs. The method was then successfully validated with respect to response function, trueness, precision, accuracy, linearity and limit of quantification. [source] Analysis of lamotrigine and its metabolites in human plasma and urine by micellar electrokinetic capillary chromatographyELECTROPHORESIS, Issue 4-5 2005Vincenzo Pucci Abstract A reliable micellar electrokinetic capillary chromatographic method was developed and validated for the determination of lamotrigine and its metabolites in human plasma and urine. The variation of different parameters, such as pH of the background electrolyte (BGE) and Sodium dodecyl sulfate (SDS) concentration, were evaluated in order to find optimal conditions. Best separation of the analytes was achieved using a BGE composed of 10 mM borate and 50 mM SDS, pH 9.5; melatonin was selected as the internal standard. Isolation of lamotrigine and its metabolites from plasma and urine was accomplished with an original solid-phase extraction procedure using hydrophilic-lypophilic balance cartridges. Good absolute recovery data and satisfactory precision values were obtained. The calibration plots for lamotrigine and its metabolites were linear over the 1,20 ,g/mL concentration range. Sensitivity was satisfactory; the limits of detection and quantitation of lamotrigine were 500 ng/mL and 1 ,g/mL, respectively. The application of the method to real plasma samples from epileptic patients under therapy with lamotrigine gave good results in terms of accuracy and selectivity, and in agreement with those obtained with an high-performance liquid chromatography (HPLC) method.* [source] DNA extraction procedure: a critical issue for bacterial diversity assessment in marine sedimentsENVIRONMENTAL MICROBIOLOGY, Issue 2 2006Gian Marco Luna Summary In order to evaluate whether different DNA extraction procedures can affect estimates of benthic bacterial diversity, based on 16S rRNA gene terminal restriction fragment length polymorphism (T-RFLP) fingerprinting technique, we compared two in situ lysis procedures (a SDS-based protocol and a commercial kit for DNA recovery) and one cell-extraction protocol on a variety of marine sediments. Despite the two in situ lysis procedures resulted in significantly different DNA yields (highest with the SDS in situ lysis), estimates of bacterial diversity provided a not significantly different ribotype richness, as well as similar values of the Shannon-Wiener (H,) and Margalef (d) indices of biodiversity and of evenness (Pielou index, J). Conversely, the cell-extraction procedure for DNA extraction resulted always in a significantly lower ribotype richness and diversity. The analysis of similarities (anosim) among the T-RFLP electropherograms allowed concluding that ribotypes composition did not change significantly using different protocols. However, the analysis of ,-diversity (turnover diversity) revealed that a large number of ribotypes was observed exclusively with one of the three protocols utilized. When unshared ribotypes from in situ lysis and cell extraction were pooled together, total ribotype richness resulted much higher (up to 80%). Our results indicate that estimates of ribotype diversity based on a single protocol of DNA extraction can significantly underestimate the total number of bacterial ribotypes present in the benthic domain. We recommend that future studies will not only integrate different DNA extraction procedures, but also will explore the possibility of integrating two or more different genetic markers in order to increase our ability to detect the actual bacterial diversity in environmental samples. [source] Comparison of the effectiveness of five extraction methods for Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum from potato tubersEPPO BULLETIN, Issue 2 2001J. Martin In the EU Control Directives, the recommended extraction procedure for testing potatoes for Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum comprises incubation followed by differential centrifugation. This method can be qualified as complex because of the number of different steps required. This study evaluates five different extraction methods for each bacterium from both a technical point of view and for the quality of the results. Results showed that in the case of C. m. sepedonicus the clarification step should be avoided. The incubation/shaking method with three subsamples gives at least as satisfactory results as the official EU procedure. It also has other advantages, facilitating immunofluorescence readings due to the reduced quantity of plant debris, and improving the speed and the reliability of the analysis. [source] Bacterial formation of phosphatic laminites off PeruGEOBIOLOGY, Issue 3 2009E. T. ARNING Authigenic phosphatic laminites enclosed in phosphorite crusts from the shelf off Peru (10°01, S and 10°24, S) consist of carbonate fluorapatite layers, which contain abundant sulfide minerals including pyrite (FeS2) and sphalerite (ZnS). Low ,34Spyrite values (average ,28.8,) agree with bacterial sulfate reduction and subsequent pyrite formation. Stable sulfur isotopic compositions of sulfate bound in carbonate fluorapatite are lower than that of sulfate from ambient sea water, suggesting bacterial reoxidation of sulfide by sulfide-oxidizing bacteria. The release of phosphorus and subsequent formation of the autochthonous phosphatic laminites are apparently caused by the activity of sulfate-reducing bacteria and associated sulfide-oxidizing bacteria. Following an extraction,phosphorite dissolution,extraction procedure, molecular fossils of sulfate-reducing bacteria (mono- O -alkyl glycerol ethers, di- O -alkyl glycerol ethers, as well as the short-chain branched fatty acids i/ai -C15:0, i/ai -C17:0 and 10MeC16:0) are found to be among the most abundant compounds. The fact that these molecular fossils of sulfate-reducing bacteria are distinctly more abundant after dissolution of the phosphatic laminite reveals that the lipids are tightly bound to the mineral lattice of carbonate fluorapatite. Moreover, compared with the autochthonous laminite, molecular fossils of sulfate-reducing bacteria are: (1) significantly less abundant and (2) not as tightly bound to the mineral lattice in the other, allochthonous facies of the Peruvian crusts consisting of phosphatic coated grains. These observations confirm the importance of sulfate-reducing bacteria in the formation of the phosphatic laminite. Model calculations highlight that organic matter degradation by sulfate-reducing bacteria has the potential to liberate sufficient phosphorus for phosphogenesis. [source] On removing the primary field from fixed-wing time-domain airborne electromagnetic data: some consequences for quantitative modelling, estimating bird position and detecting perfect conductorsGEOPHYSICAL PROSPECTING, Issue 4 2001Richard Smith In the process of removing the primary field from fixed-wing time-domain airborne EM data, the response is decomposed into two parts, which are referred to here as the time-domain ,in-phase' and ,quadrature' components. The time-domain in-phase component is dominated by the primary field, which varies significantly as the transmitter,receiver separation changes. The time-domain quadrature component comes solely from the secondary response associated with currents induced in the ground and this is the component that has traditionally been used in the interpretation of data from fixed-wing towed-bird time-domain EM systems. In the off-time, the quadrature response is very similar to the total secondary response. However, there are large differences in the on-time and even some small differences in the off-time.One consequence of these differences is that when airborne EM data are to be interpreted using a synthetic mathematical model, the synthetic data calculated should also be the quadrature component. A second consequence relates to the time-domain in-phase component which is sometimes used to estimate the receiver-sensor (bird) position. The bird-position estimation process assumes there is no secondary field in the in-phase component. If the ground is resistive, the secondary contained in the in-phase component is small, so the bird-position estimate is accurate to about 30 cm, but in highly conductive areas the secondary contribution can be large and the position estimate can be out by as much as 5 m. A third consequence arises for highly conductive bodies, the response of which is predominantly in-phase. This means that any response from these types of body is lost in the component that has been removed in the primary-field extraction procedure. However, if the bird position is measured very accurately, the actual free-space primary field can be estimated. If this is then subtracted from the estimated primary (actually free-space primary plus secondary in-phase response), then the residual is the secondary in-phase response of the ground. Using this methodology, very conductive ore bodies could be detected. However, a sensitivity analysis shows that detection of a large vertically dipping very conductive body at 150 m depth would require that the bird position be measured to an accuracy of about 1.4 cm and the aircraft attitude to within about 0.01°. Such tolerances are very stringent and not easily attainable with current technology. [source] Simultaneous Determination of Fluorine, Chlorine, Bromine and Iodine in Six Geochemical Reference Materials Using Pyrohydrolysis, Ion Chromatography and Inductively Coupled Plasma-Mass SpectrometryGEOSTANDARDS & GEOANALYTICAL RESEARCH, Issue 4 2009Hélène Balcone-Boissard halogènes; pyrohydrolyse; chromatographie ionique; spectrométrie de masse couplée à une source de plasma induit; matériaux géologiques de référence Concentrations of halogens (fluorine, chlorine, bromine and iodine) were determined in six geochemical reference materials (BHVO-2, GS-N, JG-1, JR-1, JB-1b, JB-2). Halogens were first extracted from powdered samples using a pyrohydrolysis technique, then hydrolysis solutions were analysed by ion chromatography for F and Cl and inductively coupled plasma-mass spectrometry for Br and I. The detection limits in solutions were 100 ,g l,1 for both F and Cl and 10 ng l,1 for Br and I. Considering the extraction procedure, performed on a maximum of 500 mg of sample and producing 100 ml of pyrohydrolysis solution, detection limits in rock samples were 20 mg kg,1 for F and Cl and 2 ,g kg,1 for Br and I. The mean analytical errors on the studied composition ranges were estimated at 10 mg kg,1 for F and Cl, 100 ,g kg,1 for Br and 25 ,g kg,1 for I. The concentration values, based on repeated (generally > 10) sample analysis, were in good agreement generally with published values and narrowed the mean dispersion around mean values. Large dispersions are discussed in terms of samples heterogeneity and contaminations during sample preparation. Basaltic RMs were found to be more suitable for studies of halogen compositions than differentiated rock material, especially granites , the powders of which were heterogeneous in halogens at the 500 mg level. Les concentrations en halogènes (fluor, chlore, brome et iode) on été déterminées dans 6 matériaux géologiques de référence (BHVO-2, GS-N, JG-1, JR-1, JB-1b, JB-2), distribués par le GSJ, l'USGS et le CRPG. Les halogènes étaient d'abord extraits des échantillons, disponibles sous forme de poudre, par pyrohydrolyse. F et Cl sont ensuite analysés par chromatographie ionique, Br et I par spectrométrie de masse couplée à une source de plasma induit. Les limites de détection sont de 100 ,g l,1 pour F et Cl, et de 10 ng l,1 pour Br et I, respectivement. L'extraction des halogènes était réalisée sur 500 mg de poudre de roche, produisant 100 ml de solution d'extraction. Ainsi, pour les échantillons de roche, les limites de détection étaient de 20 mg kg,1 pour F et Cl, et 2 ,g kg,1 pour Br et I. L'erreur analytique moyenne sur la gamme de concentration étudiée est estimée à 10 mg kg,1 pour F et Cl, 100 ,g kg,1 pour Br et 25 ,g kg,1 pour I. Les valeurs de concentrations données, obtenues par l'analyse répétée (> 10) du même échantillon étaient en accord avec les valeurs reportées dans la littérature. Elles présentent en général une plus faible dispersion autour de la valeur moyenne. Dans le cas d'une importante dispersion des résultats, celle-ci est discutée en terme d'hétérogénéité de l'échantillon analysé et de contamination durant la préparation du matériel de référence. Les échantillons de référence de composition basaltique se révèlent être plus appropriés pour étudier les compositions en halogènes que les matériaux correspondant à des roches différenciées, en particulier des granites dont la distribution en halogènes apparaît hétérogène dans les poudres à l'échelle d'un aliquot de 500 mg. [source] Initial evaluation of a field-friendly extraction procedure for the enzymatic assay of cassava cyanogensINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 8 2007Gerard M. O'Brien Summary A novel ,field friendly' extraction procedure has been developed for the enzymatic colorimetric determination of cyanogenic potential (CNP) in fresh cassava root parenchyma. The novel procedure does not require electrical power or vacuum, and employs inexpensive lightweight equipment, making it suitable for remote field sites. Testing of the procedure involved ten fresh roots (24,80 mg kg,1 total CNP, as HCN, fresh basis). From the parenchyma of each root, one extract was made using the novel procedure, and a ,control' extract was made using a traditional laboratory-based procedure. Total CNP assay of the extracts indicated strong (y = mx) or very strong (y = mx + c) correlation of results obtained using the two procedures, while a very strong correlation (y = mx) was obtained for free HCN. Based on this preliminary evidence, the novel procedure is satisfactory at least for total CNP assay of fresh low-CNP cassava roots. [source] Detection of hazelnut DNA traces in chocolate by PCRINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 6 2003Lieve Herman Summary By use of the DneasyTM Plant Tissue kit (Qiagen Inc.) plant DNA could be extracted from chocolate and related matrices. The polymerase chain reaction (PCR) detection of mitochondrial plant DNA is directly correlated with the length of the amplified fragment indicating shearing of DNA during chocolate production. Hazelnut DNA could be specifically detected in chocolate matrices with primers derived from the intron between exon B and C of the mitochondrial gene nad1. Specificity was confirmed towards individual chocolate ingredients and in 20 hazelnut negative chocolates. From taxonomically closely related plant species, only Carpinus turczaninovii, Ostrya carpinifolia and Corylus americana showed cross reaction, this was because of the identical sequence of the nad1 fragment. Application of extra MgCl2 throughout the DNA extraction procedure and of a specially designed Mg2+ buffered PCR, increased the detection sensitivity of co-processed hazelnut in chocolate to 0.001% or 10 ppm. [source] An optimized method to separate reticulocytes from peripheral blood for molecular analysisINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 3 2009R. PETRUZZELLI Summary A method based on immunomagnetic sorting of reticulocytes from peripheral blood was set up and combined to a commercial extraction kit for the isolation of total RNA from whole blood. This procedure resulted in high-quality RNA samples suitable for molecular analysis. We used this procedure to analyse erythroid-specific transcripts, starting from peripheral blood samples, to search for differently expressed mRNAs in patients with hereditary persistence of foetal haemoglobin. After erythrocyte lysis, CD15+and CD45+ peripheral cells were negatively sorted to remove leucocyte populations that could have affected the subsequent screening procedure. The cell sorting and RNA extraction procedure was completed within 1,2 h of erythrocyte lysis, which represents a consistent saving of time compared with other procedures. Moreover, it produced 1 ,g of total RNA per ml of blood samples, which is sufficient for molecular analysis. Therefore, our method is a reliable and efficient tool to isolate RNA from specific cell subpopulations poorly represented in peripheral blood, particularly when accurate detection and characterization of highly unstable and poorly expressed molecules is required. [source] Model explaining the predisposition to donate blood from the social marketing perspectiveINTERNATIONAL JOURNAL OF NONPROFIT & VOLUNTARY SECTOR MARKETING, Issue 3 2009Asunción Beerli-Palacio The purpose of this research is to develop a model of the explanatory factors that determine the predisposition to donate blood in order to improve the effectiveness of donor recruitment and retention programs. A personal survey was conducted on a sample of 303 potential donors between 18 and 60 years old and from both sexes, who are resident in Las Palmas de Gran Canaria (Spain) and have either never donated blood or not donated in the last 3 years. The findings lead us to say that the predisposition to donate blood is positively influenced by the information that the potential donor has about the requirements to become a donor, and by the motivations to donate blood. It is negatively influenced by the inhibiting factor of fear of the extraction procedure and its after-effects. However, prior experience as a donor and links with reference groups who are donors do not have any direct influence on the predisposition. These findings suggest that it is necessary (1) to design communication campaigns in which information and education are the goals, and which aim to make donation a habit; (2) to clarify to society the need for blood donation and to describe the process of donation in order to eliminate those inhibitors linked to fear and the perception of risks; (3) to design advertising campaigns focused on rational messages since information exercises a greater influence on the predisposition to donate than motivations linked to altruism; (4) to recommend that no great efforts be made to recapture previous donors, since experience is not a direct antecedent of the predisposition to donate but an indirect antecedent via information and (5) to stimulate word-of-mouth among reference groups using member-get-member programs whereby current donors bring new donors to the system. Copyright © 2008 John Wiley & Sons, Ltd. [source] New RF extrinsic resistances extraction procedure for deep-submicron MOS transistorsINTERNATIONAL JOURNAL OF NUMERICAL MODELLING: ELECTRONIC NETWORKS, DEVICES AND FIELDS, Issue 2 2010J. C. Tinoco Abstract The modeling of MOS transistors used for RF applications needs the definition of a lumped equivalent circuit where the intrinsic device and series extrinsic resistances are properly evaluated. The model accuracy depends on the extraction precision of each intrinsic lumped element. In order to determine the intrinsic device behavior, it is necessary to first remove the series extrinsic resistances. For this reason their extraction becomes critical for the modeling of MOS transistors in RF circuit design. Several extraction methods have been proposed; nevertheless, the measurement noise strongly affects the obtained results. The method proposed by Bracale and co-workers is the most robust extraction procedure against measurement noise, but fails to predict correctly the series extrinsic resistances for deep-submicron devices. For those reasons, we deeply analyze the method proposed by Bracale in order to understand and then overcome its limitations. Based on those analyses, a robust extraction method for deep-submicron devices is proposed. Copyright © 2009 John Wiley & Sons, Ltd. [source] A novel approach to extract accurate design parameters of PiN diodeINTERNATIONAL JOURNAL OF NUMERICAL MODELLING: ELECTRONIC NETWORKS, DEVICES AND FIELDS, Issue 6 2007Tarek Ben Salah Abstract Accurate modelling of PiN diode transient behaviour is necessary to extract design parameters which are not documented in datasheets. To meet this requirement, this paper introduces a novel approach giving the possibility to identify accurate parameters of a given device. The used technique is based only on two stages. First, the design parameters are initialized and optimized. Second, they are refined by minimizing the cost function which depends on the transient switching parameters (IRM, VRM and trr). With a simple and CPU time-saving approach this technique leads to extract design parameters without necessarily knowing the exact technological architecture of the PiN diode. Moreover, in order to validate the proposed approach and the parameter extraction procedure three commercial diodes are tested. A good agreement between experimental and simulation data is obtained. Copyright © 2007 John Wiley & Sons, Ltd. [source] Accurate substrate modelling of RF CMOSINTERNATIONAL JOURNAL OF NUMERICAL MODELLING: ELECTRONIC NETWORKS, DEVICES AND FIELDS, Issue 3 2006M. S. Alam Abstract The losses within the substrate of an RF IC can have significant effect on performance in a mixed signal application. In order to model substrate coupling accurately, it is represented by an RC network to account for both resistive and dielectric losses at high frequency (> 1 GHz). A small-signal equivalent circuit model of an RF IC inclusive of substrate parasitic effect is analysed in terms of its y -parameters and an extraction procedure for substrate parameters has been developed. By coupling the extracted substrate parameters along with extrinsic resistances associated with gate, source and drain, a standard BSIM3 model has been extended for RF applications. The new model exhibits a significant improvement in prediction of output reflection coefficient S22 in the frequency range from 1 to 10 GHz in device mode of operation and for a low noise amplifier (LNA) at 2.4 GHz. Copyright © 2006 John Wiley & Sons, Ltd. [source] A new procedure for nonlinear statistical model extraction of GaAs FET-integrated circuitsINTERNATIONAL JOURNAL OF RF AND MICROWAVE COMPUTER-AIDED ENGINEERING, Issue 5 2003Francesco Centurelli Abstract A new statistical nonlinear model of GaAs FET MMICs which allows the representation of distance-dependent technological parameter variations by means of equivalent circuit parameters, and an automatic extraction procedure, are presented. The capability to reproduce statistical distribution has been successfully checked on S parameters at different distances in the 1,50 GHz frequency range. © 2003 Wiley Periodicals, Inc. Int J RF and Microwave CAE 13, 348,356, 2003. [source] Reproductive stem cell research and its application to urologyINTERNATIONAL JOURNAL OF UROLOGY, Issue 2 2008Takehiko Ogawa Abstract: Germ cells are defined by their innate potential to transmit genetic information to the next generation through fertilization. Males produce numerous sperm for long periods to maximize chances of fertilization. Key to the continuous production of large numbers of sperm are germline stem cells and their immediate daughter cells, functioning as transit amplifying cells. Recently, it has become possible to expand germline stem cells of rodents in vitro. In addition, multipotent stem cells, which are functionally the same as embryonic stem cells, have been established from neonatal mouse testes. These stem cells derived from the testis should contribute to biological research and technologies. On the other hand, the nature of human spermatogenesis is largely unknown due to the lack of an appropriate experimental system. However, the prevailing testicular sperm extraction procedure unraveled hitherto unknown facets of human spermatogenesis. The establishment of a culturing method for human spermatogonial stem cells in hopefully the near future would be a great benefit for achieving further insight into human spermatogenesis and should lead to more sophisticated diagnostic and therapeutic clinical measures for male infertility. [source] Liquid,liquid extraction of Hg(II) from acidic chloride solutions using bis-2-ethylhexyl sulfoxideJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 7 2001Tania Francis Abstract The liquid,liquid extraction of Hg(II) from acidic chloride solutions has been studied using bis-2-ethylhexyl sulfoxide (B2EHSO) as an extractant. For comparison, extraction studies have also been carried out using di- n -octyl sulfoxide (DOSO) and diphenyl sulfoxide (DPhSO). The extraction data have been analysed by both graphical and theoretical methods taking into account aqueous phase speciation and all plausible complexes extracted into the organic phase. These results demonstrate that Hg(II) is extracted into xylene as HgCl2.3R2SO (where R2SO represents the sulfoxide). The equilibrium constant of the extracted complex has been deduced by non-linear regression analysis. The developed liquid,liquid extraction procedure has been applied for the recovery of mercury from the brine-sludge of a Chlor-Alkali plant. © 2001 Society of Chemical Industry [source] Development of an Antibody Hapten-Chip System for Detecting the Residues of Multiple Antibiotic Drugs,JOURNAL OF FORENSIC SCIENCES, Issue 4 2009Ailiang Chen M.Sc. Abstract:, The abuse of antibiotic drugs during animal production remains a worldwide problem and the subsequent detection of the residues of various drugs present at low concentrations in complex biological matrices poses significant analytical challenges. The present study outlines a practical biochip assay system to identify antibiotic residues in different animal tissue extracts. The system uses a simple but efficient multiresidue sample extraction procedure to isolate the antibiotic residues which were then identified directly using high-affinity monoclonal antibodies presented in a competitive immunoassay with conjugated antibiotic hapten-chips. The hapten-chip can analyze six samples each for eight antibiotics on a single chip within 3 h. The analytical results with both artificial positive standard samples and the incurred samples show that the antibody hapten-chip system has a comparable accuracy and a similar sensitivity to a standard ultra performance liquid chromatography,mass spectrometry (MS)/MS assay. In conclusion, an effective analytical screening system based on antibody hapten-chip was developed for detecting multiple antibiotic residues from multiple samples. [source] An improved independent component regression modeling and quantitative calibration procedureAICHE JOURNAL, Issue 6 2010Chunhui Zhao Abstract An improved independent component regression (M-ICR) algorithm is proposed by constructing joint latent variable (LV) based regressors, and a quantitative statistical analysis procedure is designed using a bootstrap technique for model validation and performance evaluation. First, the drawbacks of the conventional regression modeling algorithms are analyzed. Then the proposed M-ICR algorithm is formulated for regressor design. It constructs a dual-objective optimization criterion function, simultaneously incorporating quality-relevance and independence into the feature extraction procedure. This ties together the ideas of partial-least squares (PLS), and independent component regression (ICR) under the same mathematical umbrella. By adjusting the controllable suboptimization objective weights, it adds insight into the different roles of quality-relevant and independent characteristics in calibration modeling, and, thus, provides possibilities to combine the advantages of PLS and ICR. Furthermore, a quantitative statistical analysis procedure based on a bootstrapping technique is designed to identify the effects of LVs, determine a better model rank and overcome ill-conditioning caused by model over-parameterization. A confidence interval on quality prediction is also approximated. The performance of the proposed method is demonstrated using both numerical and real world data. © 2009 American Institute of Chemical Engineers AIChE J, 2010 [source] MALDI-TOF mass spectrometry of hordeins: rapid approach for identification of malting barley varietiesJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 9 2009alplachta Abstract A procedure for identification of malting barley varieties using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) of ethanol-soluble barley proteins (hordeins) is described. The hordeins were first extracted from milled barley grains by several extraction protocols (using different extraction agents and conditions). Hordein extracts were then analyzed directly via MALDI-TOF MS without any preliminary purification or separation step, and the protein profiles of analyzed hordein extracts were compared in order to find out the most suitable extraction procedure for mass spectrometric analysis. The optimized procedure was successfully applied to identification of 13 malting barley varieties. Our results revealed that the proposed mass spectrometry-based approach provides characteristic mass patterns of extracted hordeins, which can be advantageously used for barley variety identification. Copyright © 2009 John Wiley & Sons, Ltd. [source] In-gel deglycosylation of sodiumdodecyl sulfate polyacrylamide gel electrophoresis-separated glycoproteins for carbohydrate estimation by matrix-assisted laser desorption/ionization time-of-flight mass spectrometryJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 3 2002S. Kilz Abstract Mass determination by mass spectrometric methods (electrospray ionization mass spectrometry (ESI-MS), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS)) of sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)-separated proteins is a well known procedure and reliable protocols are available. In our efforts to use the established methods to determine the molecular mass of the disulfide brigded, heterodimeric glycoprotein GP3 and to determine the carbohydrate content of each protein subunit we developed an in-gel chemical deglycosylation method. For this purpose we established experimental conditions that allow maximum extraction of the high molecular mass protein subunits and developed a routine method to apply the HF,pyridine deglycosylation protocol to proteins isolated from polyacrylamide gel pieces. The novel protocol and extraction procedure described can be used to analyze O -glycosylated proteins up to 150 kDa after SDS-PAGE separation. Copyright © 2002 John Wiley & Sons, Ltd. [source] Novel selective cytotoxicity of wild sarsaparilla rhizome extractJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 10 2006Y. G. Huang Among six fractions, including total extract and fractions of hexane, ethyl acetate, butanol, water and boiling water extracted and separated from wild sarsaparilla rhizome, the hexane fraction (HRW) was the most effective in eliminating four different human cancer cell lines with cellular viability less than 6.8%. HRW exhibited the highest potency against human leukaemia cells with an IC50 (concentration that inhibited the growth rate of cells by 50%) of 3.3 ± 0.3 ,g mL,1, which was 17.6-fold smaller than that against normal human umbilical vein endothelial cells (IC50, 58.0 ± 1.5 ,g mL,1). For its rich natural resources, simple extraction procedure and high yield (3.2%), HRW has the potential to be developed as a selective anti-cancer nutraceutical or pharmaceutical natural health product with low side effects and high economical return. [source] Body Distribution of Poly- DL -lactide-poly(ethylene glycol) Microspheres with Entrapped Leptospira interrogans Antigens Following Intravenous and Oral Administration to Guinea-pigsJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 7 2000XIAOHONG LI Poly- DL -lactide-poly(ethylene glycol) (PELA) microspheres with entrapped antigens were administered intravenously and orally into guinea-pigs to quantitatively determine the in-vivo distribution and release profiles. PELA microspheres containing 125I-labelled outer-membrane protein Leptospira interrogans antigens (125I-OMP) were prepared by double-emulsion solvent extraction procedure, and characterized with respect to size, morphology and in-vitro release profiles. The fractured sections of liver and spleen were inspected by scanning electron microscopy, which indicated that microspheres had successfully been entrapped within the above tissues after intravenous injection and oral administration. At predetermined intervals, the blood and such tissues as the liver, spleen, kidney, thyroid, small intestine and mesentery were collected, and the radioactivity was measured by gamma scintillation counting. Following intravenous administration, 56.7% of administered microspheres were accumulated in immunization-related tissues, and 40.1% of microspheres were located in the liver and spleen. However, there was limited uptake efficiency (8.33%) following oral administration, and 49.5% of the absorbed microspheres were located in the intestinal mucosa. Compared with in-vitro release, the in-vivo release profiles of 125I-OMP from PELA microspheres, determined from the decreasing radioactivity in the above tissues, were much faster and the burst effect was higher. Antigen-loaded PELA microspheres were efficiently entrapped within immunization-related tissues after intravenous administration, but orally administered PELA microspheres showed limited uptake efficiency. Further investigation is needed to improve intestinal absorption. [source] | ||||||||||||||||||||||||||||||||||||||||