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Detection of simple radially symmetric targets: further evidence for the matched filter processing scheme in human pattern detection.

The detection of small radially symmetric targets was studied using a subthreshold summation paradigm. Small disc and disc-like patterns with diameters up to 0.60 were used for superposition on Bessel functions of zero order, subthreshold contrast and various spatial frequencies. Contrast interrelation functions prove linear over the whole range of contrasts used for the Bessel functions while their slopes show systematic variation with spatial frequency. An extrapolation of sensitivity from the slopes reveals that sensitivity can be predicted by a simple model assuming detection to be mediated by a transfer function made up as a cascade of an even bandpass function and the disc pattern spectrum, as has been found previously using one dimensional luminance distributions. Problems concerning the formation of pattern-specific radial symmetric filters are discussed.

Contrast Sensitivity↗

Rapid immunochromatographic malarial antigen detection unreliable for detecting Plasmodium malariae and Plasmodium ovale.

In order to determine the reliability of two commercial tests for the rapid detection of plasmodial antigen in cases of infection with Plasmodium ovale and Plasmodium malariae, the products were evaluated in four centers and a search of the relevant literature was performed. The results of the present and previous studies were compared. With overall sensitivities ranging between 18.8% and 47.6% for Plasmodium malariae and between 20% and 31.3% for Plasmodium ovale, it is evident that neither test is reliable for the detection of Plasmodium ovale and Plasmodium malariae infections.

Animals↗

Chemiluminescent nucleic acid detection with digoxigenin-labeled probes: a model system with probes for angiotensin converting enzyme which detect less than one attomole of target DNA.

Digoxigenin-labeled nucleic acid probes for angiotensin-converting enzyme were prepared by the polymerase chain reaction. In a model system, these probes were used to optimize conditions for target detection by direct chemiluminescence after dot-blot, Southern, or Northern hybridizations. Digoxigenin-labeled probes detected 23 fg (1.6 X 10(-19) mol or 9 X 10(4) copies) of target consistently.

Animals↗

Anthranilic acid metabolism in the isolated perfused rat liver: detection and determination of anthranilic acid and its related substances using high-performance liquid chromatography with electrochemical detection.

In order to elucidate the anthranilic acid metabolism in animal tissue, the metabolism was studied in the isolated perfused liver of rats. A sensitive and rapid method was devised for determination of anthranilic acid and its related substances using high-performance liquid chromatography with electrochemical detection. 5-Hydroxyanthranilic acid and anthranilamide, which have not been detected in animal tissue, were found in the perfusate and in the bile secreted from the perfused liver, respectively. In addition, a non-enzymatic production of anthranilamide from anthraniloyl glucuronide in the presence of ammonium and bicarbonate ions was also observed. These present results suggest that, apart from undergoing glycine or glucuronide conjugation, anthranilic acid is metabolized to 5-hydroxyanthranilic acid and anthranilamide in the rat liver.

Animals↗

Femtogram detection limits for biogenic amines using microbore HPLC with electrochemical detection.

Detection limits for biogenic amines, serotonin (5-HT), 5-hydroxyindoleacetic acid, 5-hydroxytryptophan, norepinephrine, epinephrine, dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC), of 50-200 fg are reported. This was accomplished using microbore HPLC with electrochemical detection. Reproducibility at the 1-5 pg level of 1.6-8.1% was obtained. The utility of this approach is demonstrated for analysis of brain tissues of 1 microgram or less.

3,4-Dihydroxyphenylacetic Acid↗

Development of a screening system for detection of somatic mutations. I. Enzyme immunoassay for detection of antibodies against specific hemoglobin determinants.

A solid-phase enzyme immunoassay for the detection of antibodies, specific for hemoglobin (Hb) is described. The application of glutaraldehyde resulted in a sensitive assay and allowed the use of urea, which is an important advantage if polypeptides not soluble in aqueous buffers are to be used. Mutation-carrying Hb chains can be purified, solubilized in urea and used in the immunoassay to monitor the purification and selection of antibodies specific for these variants. Specific antibodies are the main tools for the development of a hemoglobin-locus mutation system for detection of potentially mutagenic environmental agents. With erythrocytes as target cells, this system permits in vivo monitoring of subjects under exposure. Conventional antibody production, however, frequently turns out to be unsuccessful. The production of monoclonal antibodies has several advantages over conventional antibody production, but a sensitive antibody screening system is essential. Because of the sensitivity and the ease with which a large panel of antibody fractions against a vast panel of Hb antigens can be examined, the described immunoassay has potential value for the screening of hybridoma cultures.

Adult↗

Detection of antibody to the PreS2 sequence of the hepatitis B virus envelope protein using an immuno-ligand assay with a silicon sensor detection system.

Sensitive immunoassays are essential for establishing the efficacy of recombinant vaccines to hepatitis B virus (HBV). These experimental vaccines include the PreS2 and S domains of the HBV envelope protein. To facilitate measurement of antibody against HBV PreS2, we employed the immuno-ligand assay with silicon sensor-based detection. Labeling of immune reagents with the haptens biotin and fluorescein allows adaptation to the immunofiltration light addressable potentiometric sensor (LAPS) system. A biotinylated monoclonal anti-PreS2 antibody and anti-PreS2 in clinical serum samples competitively bind in liquid phase to a fluorescein labeled PreS2 + S antigen. Streptavidin mediates the immobilization on biotinylated nitrocellulose membranes. Fluorescein mediates binding of an anti-fluorescein urease conjugate to the immune complex. Urease serves as the signal-generating component which subsequently is measured in the LAPS reader. In comparison to a competitive RIA, the immuno-ligand assay demonstrated a four-fold improved sensitivity using a smaller sample volume. The higher sensitivity resulted in earlier detection of seroconversion during a clinical vaccine study.

Antibodies, Monoclonal↗

Influence of visual blur on object-motion detection, self-motion detection and postural balance.

We have compared the influence of visual blurring (either by use of plastic foils or by plus lenses) on detection of object-motion and self-motion, and on visual control of postural balance. Three experiments were performed--in all of which object-motion and self-motion detection latencies (response times) and postural sway (sway path) increased with increasing visual blurring. We conclude that visual blurring either by foils or by glasses affects object- and self-motion perception and postural sway in a similar way.

Attention↗

A comparison of the antigen detection ELISA and parasite detection for diagnosis of Trypanosoma evansi infections in camels.

Two herds of 60 camels each, living in Trypanosoma evansi endemic areas, were selected and studied for a period of 18 months. Animals in one herd were treated prophylactically with quinapyramine prosalt (May and Baker, Dagenham, UK), while those in the other herd were treated individually with quinapyramine dimethylsulphate (May and Baker, Dagenham, UK) when proven parasitaemic. The herd on prophylaxis was sampled for antigen and patent infection monthly. The other herd was sampled weekly for patent infection and fortnightly for antigen. The results obtained could be divided into four categories. The first category comprised cases (52 out of 61) in which the presence of trypanosome antigens could be correlated with parasitological diagnosis. In 80% of these animals the antigens disappeared from the circulation within a period of 30 days following chemotherapy. The second category comprised those animals with parasitologically proven infections but which did not have antigens in their sera. This was observed in nine camels, seven of which were from the herd that was being examined weekly for the presence of trypanosomes. These were considered to be animals in early infection, as the subsequent sera were also negative for anti-trypanosome antibodies and immune complexes. The third category comprised camels which were antigen-positive but aparasitaemic. Sera from these animals were also positive for anti-trypanosome antibodies, indicating that antigen-positivity was a true reflection of trypanosome infections in these animals. The last category comprised pre-weaned camel calves which appeared to have some form of protection against trypanosomiasis, as evidenced by the absence of trypanosomes, antigens and antibodies throughout the early period of their lives. Only occasional antigenaemia was found in a few calves. It is concluded that trypanosome antigen detection may give a more accurate idea of the prevalence of T. evansi infections than does whole parasite detection.

Animals↗

Spatially resolved detection of antibody-antigen reaction on solid/liquid interface using total internal reflection excited antigen fluorescence and charge-coupled device detection.

Spatially-resolved detection of antibody-antigen reactions at the solid/liquid interface was investigated by total internal reflection excited fluorescence from large area flat surfaces. Anti-HSA immunoglobulin G (IgG) antibody was immobilized at four spatially distinct spots. Binding of fluorescein-labeled human serum albumin (HSA) from the solution to immobilized antibody was detected by a cooled charge-coupled device (CCD) as a charge in the fluorescence intensity. A two-dimensional representation of the fluorescence was obtained during the binding reaction time of 25 mins. The contributions from bound and free antigen to the total signal were evaluated. The influence of the scattered excitation light and the normalization of fluorescence signal with respect to the two-dimensional incident light intensity distribution are discussed.

Antigen-Antibody Reactions↗

Comparing the ESA HPLC total homocysteine assay with electrochemical detection to the CDC in-house HPLC assay with fluorescence detection.

BACKGROUND: Environmental science has developed a simple high performance liquid chromatography (HPLC) assay with electrochemical detection for total homocysteine (tHcy) measurement that does not require derivatization of free thiols. We evaluated this method and compared it with the CDC HPLC assay with fluorescence detection (FD). METHODS: tHcy is measured after reduction of disulfides/protein-bound thiols and protein precipitation using four channels of an ESA CoulArray detector. L-homocystine is used as calibrator, penicillamine as internal standard. RESULTS: Aqueous calibration of the ESA assay resulted in overestimation of tHcy by approximately 30% compared to the HPLC-FD method. Calibration in plasma alleviated the matrix effect. The within- (n=3) and between-run (n=20) imprecision was <6%, the linearity up to 100 micromol/l was excellent, and the recovery of tHcy added to plasma was nearly complete (98.7%+/-2.3%). Good correlation was observed between both methods for 266 plasma samples. The ESA assay showed a minimal negative bias of 0.28 micromol/l (3.3%). CONCLUSION: The ESA tHcy assay performed well in terms of accuracy and precision, and showed good agreement with the CDC HPLC-FD assay when calibrated in plasma. The major advantage of this assay is that it does not require sample derivatization. Disadvantages include instability of the prepared samples for prolonged storage and matrix effects.

Calibration↗

Application of high-performance liquid chromatography with ultraviolet diode array detection and refractive index detection to the determination of class composition and to the analysis of gasoline.

A method of effective application of normal-phase high-performance liquid chromatography (NP-HPLC) with ultraviolet diode array detection (DAD) and refractive index detection (RID) for the determination of class composition of gasoline and its components, i.e. for the determination of content of alkenes, aromatic and saturated hydrocarbons in gasoline meeting modern quality standards, has been developed. An aminopropyl-bonded silica stationary phase was used along with n-hexane or n-heptane as the mobile phase. A DAD signal integrated over the 207-240 nm range was used to determine alkenes. This eliminates the necessity of separating alkenes from saturates, because the latter do not absorb UV radiation above 200 nm. The content of aromatic hydrocarbons is determined by means of a refractive index detector. Calibration was based on hydrocarbon type composition determined by the fluorescent indicator adsorption method, ASTM D1319. The results obtained by the developed method were found to be consistent with those obtained by fluorescent indicator adsorption or by a multidimensional GC method (PIONA) (ASTM D5443). The method can be applied to gasoline meeting recent quality standards, irrespective of refining technology used in the production of gasoline components, including gasoline with various contents of oxygenates. The developed method cannot be used to determine the hydrocarbon type composition of gasoline that contains as a component the so-called pyrocondensate, i.e. the fraction with a boiling point up to 220 degrees C, obtained through thermal pyrolysis of distillation residues of crude oil or coal and, consequently, does not meet the quality standards. The paper includes the procedure for identification of this type of gasoline.

Chromatography, High Pressure Liquid↗

Detection and identification of phenazone-type drugs and their microbial metabolites in ground and drinking water applying solid-phase extraction and gas chromatography with mass spectrometric detection.

A new analytical method applying in situ derivatization was developed to enable the extraction of polar drug metabolites from water samples by solid-phase extraction (SPE). An additional derivatization by silylation was used to enhance the sensitivity of analyte detection by gas chromatography-mass spectrometry (GC-MS). Thus, the two metabolites 1,5-di-methyl-1,2-dehydro-3-pyrazolone (DP) and 4-(2-methylethyl)-1,5-dimethyl-1,2-dehydro-3-pyrazolone (PDP), postulated for the degradation of phenazone and propyphenazone, were identified and detected up to the microg/L level in raw and drinking water samples from public water supply.

Acetylation↗

Identification and quantification of (polymeric) hindered-amine light stabilizers in polymers using pyrolysis-gas chromatography-mass spectrometry and liquid chromatography-ultraviolet absorbance detection-evaporative light scattering detection.

Direct analysis of polymers containing polymeric hindered amine light stabilizers (HALS) by using pyrolysis coupled to GC-MS is applied successfully for fast and straightforward identification of these HALS additives. Each of the HALS additives shows different pyrolysis gas chromatograms containing characteristic pyrolysis products. As a result, HALS additives with very similar chemical structures, e.g. Chimassorb 944 and Chimassorb 2020, can be distinguished. A HPLC method with both ultraviolet (UV) and evaporative light scattering detection (ELSD) is developed to quantify the various HALS additives in extracts of polymers. The critical factor of the HPLC method is the use of a basic amine, like n-hexylamine, as a solvent additive to facilitate the elution of HALS additives. The various HALS additives can be distinguished according to retention time and peak shape and by using different detection methods. The suitability of the developed methods is demonstrated by the analytical performance of the HPLC method and the identification and determination of the actual content of HALS additives in polyolefines using pyrolysis GC-MS and HPLC. The HPLC method can also be used for the determination of the specific migration of HALS additives from food contact materials.

Amines↗

Comparison of voltammetric detection assisted by multivariate curve resolution with amperometric detection in liquid chromatographic analysis of cysteine-containing compounds.

A voltammetric detection mode (VD) in conjunction with multivariate curve resolution with alternating least squares (MCR-ALS) method is applied to the analysis of cysteine-containing compounds and compared with a well established amperometric detection (AD) mode in a thin-layer dual Hg/Au cell. VD-MCR-ALS provides an increase in selectivity for cases where satisfactory separation of electroactive compounds is not allowed. However, concentrations needed for a good quantification in VD are higher than in AD due to much large contribution of background in VD.

Cysteine↗

Automatic program for peak detection and deconvolution of multi-overlapped chromatographic signals part I: peak detection.

A series of two papers describing a procedure for automated peak deconvolution is presented. The goal is to develop a package of routines that can be used by non-experienced users. Part I (this paper) concerns peak detection, whereas Part II is dedicated to the deconvolution itself. In this first part, the most interesting features of the peak detection algorithms, which precede the deconvolution step, are outlined. High-order derivatives provide valuable information to assess the number of underlying compounds under a given peak cluster. A smoothing technique was found essential to compute properly the derivatives, since the noise is amplified when differences are calculated. The Savitsky-Golay smoother was applied in combination with the Durbin-Watson criterion to automate the window size selection. This strategy removed the noise without loosing valuable information. In some cases, it was found preferable to split the chromatogram in different elution regions, and apply the Durbin-Watson test and the Savitsky-Golay smoother to each region, separately. The derivatives allowed obtaining estimates of both peak parameters and the corresponding ranges for each eluting compound to be used in the deconvolution. An algorithm oriented to compare peaks from different chromatograms is also presented to perform deconvolution, using information from several related chromatograms.

Algorithms↗

A comparison of a gas chromatographic with electron-capture detection and a gas chromatographic with mass spectrometric detection screening methods for the analysis of famoxadone in grapes and wines.

Famoxadone is a recent oxazolidinedione fungicide widely used in viticulture and in Integrated Pest Management strategies. In this work, after a simple and fast liquid-liquid extraction (LLE), two new gas chromatographic methods were developed to analyze famoxadone residues in grapes and wines, one with electron-capture detection (GC-ECD) and the other with mass spectrometry (GC-MS). Global uncertainties for validation parameters of both methods were compared. Limits of detection (LODs) were 0.06 and 0.02 mg/L, precision was not above 11.7 and 6.8% and recoveries were, on average, 103%+/-12 and 96%+/-12, respectively, for the GC-ECD and GC-MS methods. Similar expanded uncertainties in the range from 0.25 to 1.00 mg/L were below 35%, with increasing values for lower levels of famoxadone. GC-MS method had a lower LOD and a lower uncertainty if compared with the GC-ECD method.

Chromatography, Gas↗

Detection, characterization and identification of crucifer phytoalexins using high-performance liquid chromatography with diode array detection and electrospray ionization mass spectrometry.

We have analyzed 23 crucifer phytoalexins (e.g. brassinin, dioxibrassinin, cyclobrassinin, brassicanals A and C) by HPLC with diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS) using both negative and positive ion modes. Positive ion mode ESI-MS appeared more sensitive than negative ion mode ESI-MS in detecting this group of compounds. A new HPLC separation method, new LC-MS and LC-MS(2) data and proposed fragmentation pathways, LC retention times, and UV spectra for selected compounds are reported.

Brassicaceae↗