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At least 1,081 records · Page 60Linked to original sources
PubMed2005

Comparison of shikimic acid determination by capillary zone electrophoresis with direct and indirect detection with liquid chromatography for varietal differentiation of red wines.

Two capillary zone electrophoretic (CZE) methods for determination of shikimic acid in Chilean red wine were developed and compared with a HPLC method. Both electrophoretic methods were carried out by using a reversed electroosmotic flow induced by trimethyl(tetradecyl)ammoniumbromide (TTAB) with indirect detection at 260 nm using p-aminobenzoic acid as a UV-absorbing co-ion or by direct detection at 213 nm. In both cases, the separation was carried out in a 50 microm I.D. uncoated capillary with an effective length of 48 cm, a negative power supply of 30 kV, using a buffer based on bis[2-hydroxyethyl]imino-tris[hydroxymethyl]methane (Bis-Tris), pH 7.0 or 7.5 and hydrodynamic injection. The chromatographic separations were carried out on a C-18 reversed phase column followed by a sulfonyl-styrene-divinylbenzene (S-DVB) ion exclusion column at 70 degrees C with H2SO4 0.02 M as isocratic mobile phase and a flow rate of 0.5 mL min(-1). The three methods allowed the quantification of shikimic acid with quantification limits between 1.0 and 12.0 mg L(-1) and precision between 7.3 and 10.1%, however, only the concentrations obtained by CZE with direct detection were statistically similar to those of HPLC. This parameter was evaluated as analytical tool to verify varietal authenticity of red wines. In all cases, the Cabernet Sauvignon wines presented higher concentrations of shikimic acid, compared with Merlot or Carmenère wines.

Chromatography, High Pressure Liquid↗
PubMed2000

[Clinical study of direct digital radiography in caries detection].

OBJECTIVE: Direct digital radiography is a computer-based image technique, the purpose of this paper is to investigate possible usage of direct digital radiography-radio visiography (RVG) in caries detection in clinic. METHODS: A human head model with teeth was used to establish the operative exposure time for RVG in vitro, then RVG images for normal and carious tooth were obtained in vivo and were compared with conventional E-speed film in image quality and diagnostic accuracy. RESULTS: The optimal exposure time for RVG image was obtained, and the exposure time was different in each tooth quarter. The acceptable rate of image quality and diagnostic accuracy of RVG was statistically equivalent to that of E-speed film (P > 0.05), however, the excellent rate of RVG was statistically superior to that of E-speed film (P < 0.05). CONCLUSION: The conventional X-ray will be substituted by RVG in caries detection in clinic.

Adolescent↗
PubMed1992

Impact of specimen handling and storage on detection of hepatitis C virus RNA.

Direct detection of hepatitis C virus (HCV) RNA in serum or plasma is useful for validating the performance of anti-HCV assays and for the discrimination of persons with persistent HCV infections from those with resolved infections. Quantitation of HCV RNA may also be useful for disease prognosis and therapeutic monitoring. Previous studies have reported detection of HCV RNA in 50 to 70 percent of blood donors who were positive on anti-HCV supplemental tests. There is concern that specimen processing and storage conditions might influence the stability, and hence the detectability, of HCV RNA. To address this concern, the rate of detection of HCV RNA by the polymerase chain reaction (PCR) using donor pilot tube sera (PTS) previously subjected to routine donor screening and supplemental testing was compared with HCV PCR results obtained with fresh-frozen plasma (FFP) derived from the same donations. All 16 anti-HCV supplemental test-positive donations evaluated were HCV RNA positive with FFP, whereas only 10 (62.5%) were positive with PTS (p = 0.024). None of 11 FFP or PTS samples from HCV enzyme immunoassay-reactive donations not confirmed by supplemental anti-HCV assays tested positive for HCV RNA. Direct comparison of sample type (serum vs. plasma) and various storage conditions using specimens from two seropositive donors showed that room-temperature storage results in marked reduction in HCV RNA signal, while replicate freezing and thawing caused a moderate reduction. These data indicate that well-controlled sample processing and storage conditions are critical to the sensitive and potentially quantitative analysis of HCV RNA.

Blood Preservation↗
PubMed2002

A dynamically modified microfluidic poly(dimethylsiloxane) chip with electrochemical detection for biological analysis.

Separation and direct detection of amino acids, glucose and peptide in a 3.1 cm separation channel made of poly(dimethylsiloxane) (PDMS) with end-column amperometric detection at a copper microdisk electrode was developed. This system is the integration of a normal sized working electrode with electrochemical detection on a PDMS microfabricated device. The PDMS channels dynamically modified by 2-morpholinoethanesulfonic acid (MES) show less adsorption and more enhanced efficiency than that of unmodified ones when applied to separations of these biological molecules. The migration time is less than 100 s and the reproducibility of migration time is satisfactory with relative standard deviation (RSD) of 2.8% in 19 successive injections. The limits of detection of arginine (Arg), glucose, and methionine-glycine (Met-Gly) are estimated to be 2.0, 8.5, and 64.0 microM at S/N = 3, approximately 0.5-16.0 fmol, respectively. Variances influencing the separation efficiency and amperometric response, including injection, separation voltage, detection potential, or concentration of buffer and additive, are assessed and optimized.

Alkanesulfonic Acids↗
PubMed1999

Biochemical characterization of lucigenin (Bis-N-methylacridinium) as a chemiluminescent probe for detecting intramitochondrial superoxide anion radical production.

Direct detection of intramitochondrial superoxide anion radical (O(-*)(2)) production is of critical importance for investigating the pathophysiological consequences resulting from altered cellular reactive oxygen homeostasis. The purpose of this study with isolated mitochondria was to characterize the biochemical basis for lucigenin as a chemiluminescent probe to detect intramitochondrial O(-*)(2) production. Incubation of isolated mitochondria with lucigenin at non-redox cycling concentration produced lucigenin-derived chemiluminescence (LDCL), which was increased markedly by mitochondrial substrates, pyruvate/malate or succinate. The LDCL was reduced greatly by the membrane permeable superoxide dismutase (SOD) mimetics, 2,2,6,6-tetramethylpiperidine-N-oxyl and Mn(III)tetrakis(1-methyl-4-pyridyl)porphyrin, but not by Cu,Zn-SOD. With an ion-pair HPLC method, a concentration-dependent accumulation of lucigenin was detected within mitochondria. The accumulation of lucigenin by mitochondria was reduced markedly in the presence of carbonyl cyanide p-(trifluoromethoxy)phenyhyldrazone, an uncoupler known to dissipate the mitochondrial membrane potential. With submitochondrial particles, we observed that both complexes I and III of the mitochondrial electron transport chain appear to be able to catalyze the one electron reduction of lucigenin, a critical step involved in LDCL. After incubation of mitochondria with lucigenin at non-redox cycling concentrations, formation of N-methylacridone, the proposed end product of the reaction pathway leading to LDCL, within the mitochondrial fraction was also detected. In addition, a significant linear correlation was observed between the LDCL and either the lucigenin accumulation or the N-methylacridone formation within the mitochondria. Taken together, our results conclusively demonstrate that when properly used LDCL can reliably detect intramitochondrial O(-*)(2) production.

Acridines↗
PubMed2004

A selective protein sensor for heparin detection.

No clinical assays for the direct detection of heparin in blood exist. To create a heparin sensor, the hyaluronan (HA)-binding domain (HABD) of a protein that binds heparin and HA was engineered. GST fusion proteins containing one to three HABD modules were cloned, expressed, and purified. The affinities of each construct for heparin and for HA were determined by a competitive enzyme-linked immunosorbent assay using immobilized HA or heparin. Each of the constructs showed modest affinity for immobilized HA. However, heparin was 100-fold more potent than HA as a competing ligand. With immobilized heparin, affinity increased as the HABD copy number increased. The three-copy construct, GST-HB3, detected unfractionated free heparin (UFH) as low as 39ng/ml (equivalent to approximately 0.1U/ml) with a signal-to-noise ratio of 5.6. GST-HB3 also showed 100-fold selectivity for heparin in preference to other glycosaminoglycans. The plot of logKd vs log [Na+] showed 2.5 ionic interactions per heparin-HB3 interaction. GST-HB3 showed a linear detection of both UFH (15kDa) and low-molecular-weight heparin (LMWH; 6kDa) added to human plasma. For UFH, the range examined was 78 to over 2000ng/ml (equivalent to 0.2 to 5.0U/ml). For LMWH, the useful range was 312 to over 2000ng/ml. The coefficient of variance for the assay was < 9% for six serial heparin dilutions and <12% for three plasma samples. In clinical use, GST-HB3 could accurately measure therapeutic heparin levels in plasma (0.2 to 2U/ml).

Amino Acid Sequence↗
PubMed2001

Infant direction discrimination thresholds.

Although adults can detect direction differences as small as 1 arc degree, the ability of infants to discriminate direction of motion is less clear. This study measures the precision with which 6-, 12-, and 18-week-old infants discriminate direction of motion. Infants viewed random dot kinematograms in which a direction difference between the target and background dots defined a circular target. The target was then placed into continuous motion. An FPL paradigm was used to assess infants' preference for the target as a function of the direction difference between the target and background dots. Direction discrimination thresholds with a moving target were indeterminate at 6 weeks of age, 22 degrees at 12 weeks of age and 17 degrees at 18 weeks of age. This precision was maintained across different testing conditions. However, performance dropped markedly when dot motion was presented within a flickering stationary target. It was concluded that infants can make relatively fine discriminations of motion direction if given an engaging stimulus.

Child Development↗
PubMedDate not supplied

Indocyanine green angiograms of choroidal nevi. comparison between confocal and nonconfocal scanning laser ophthalmoscope and fundus video camera.

PURPOSE: A fundus video camera and a nonconfocal scanning laser ophthalmoscope (SLO) detect direct light and indirect light, whereas a confocal SLO detects mostly direct light. Differences in confocal and nonconfocal SLO images and fundus video camera images are most likely due to their different optical systems. These differences were examined in indocyanine green (ICG) angiograms of a choroidal nevus. METHODS: A confocal SLO, a nonconfocal SLO, and a high resolution digital fundus video camera were used to obtain ICG angiograms of pigmented choroidal nevi in 4 patients for 30 minutes following dye injection. RESULTS: All the angiograms showed a hypofluorescent region in the nevus until 10-14 minutes after dye injection, except in 1 patient in whom no hypofluorescent region was seen in an early confocal-SLO angiogram. From 20 minutes to 30 minutes postinjection, the hypofluorescent regions were still visible in all fundus video camera angiograms and nonconfocal SLO angiograms but not in confocal SLO angiograms. CONCLUSIONS: Early angiograms taken with the three angiography systems showed a similar appearance of the choroidal nevus. However, late ICG angiograms with a confocal SLO showed different images from those taken with a nonconfocal SLO or a fundus video camera. It is suggested that the angiography system and the aperture size of an SLO should be selected according to the aspect of the pigmented choroidal nevus that is of interest in late-phase ICG angiography.

Adult↗
PubMed2003

A comparison of methods for detecting adenovirus type 8 keratoconjunctivitis during a nosocomial outbreak in a Neonatal Intensive Care Unit.

BACKGROUND: An outbreak of epidemic keratoconjunctivitis (EKC) due to adenovirus (Ad) type 8 and involving 14 members of the hospital staff and 33 neonates admitted to the Neonatal Intensive Care Unit of the local University Hospital occurred between September and December 2000 in Pavia, Italy. The outbreak was preceded by an outbreak of EKC within the community. OBJECTIVE: To compare the performance of conventional virus isolation on cell cultures, direct detection of Ad antigens in conjunctival cells by a direct fluorescent assay (DFA) and Ad DNA detection in conjunctival swabs by polymerase chain reaction (PCR) for diagnosis of adenoviral conjunctivitis. STUDY DESIGN: Of conjunctival swabs collected from 47 patients, all were tested by virus isolation, 43 by direct Ad antigen detection, and 37 by Ad DNA detection. Direct Ad antigen detection was carried out by DFA using a group-specific monoclonal antibody. Detection and subgrouping of Ad DNA by nested PCR was performed using two sets of primers complementary to hexon and fiber genes, respectively. RESULTS: Ad was detected in 24/47 (51.1%), 21/43 (48.8%), and 23/37 (62.1%) samples by virus isolation, direct antigen detection and PCR, respectively. Overall, 30/47 (63.8%) samples were Ad-positive. Of 37 specimens tested in parallel by all three methods, Ad was detected by at least one of the three techniques in 26/37 (70.3%). All Ad isolates were identified as serotype 8 by neutralization, while all PCR-positive samples were identified as belonging to subgroup D. No other virus was isolated from any conjunctival swab. Time required for test completion was 9.6 (4-20) days for virus isolation, 1-2 h for DFA and 24 h for PCR. CONCLUSIONS: DFA was a sensitive and rapid assay but results depend on the quality of sample and the expertise of the observer. PCR was the most sensitive assay, although it takes longer to perform and requires dedicated facilities; thus, it could be restricted to DFA-negative samples. Virus isolation is still useful from an epidemiological point of view.

Adenovirus Infections, Human↗
PubMed2006

[Determination of tetracyclines by a new spectrum technique].

Tetracyclines are light-fluorescence substances, which cannot be detected directly by fluorometry. Herein a new spectrum method was proposed to detect tetracyclines directly by fluorometry. Under optimal conditions, the calibration graph is linear over the range 0.10-9.00 microg x mL(-1) for tetracyclines, and the detection limits of tetracycline, oxytetracycline, chlortetracycline and doxycycline are 0.065, 0.067, 0.068 and 0.070 microg x mL(-2), respectively.

Anti-Bacterial Agents↗
PubMed2005

Direct bidirectional angle-insensitive imaging of the flow signal intensity in Doppler optical coherence tomography.

We introduce a new method, to our knowledge, for direct detection of flow signal intensity by stationary target rejection. In our system, two delay lines are constructed with identical scanning speed and ranging depth. One delay line is used for depth ranging as well as phase modulation, and the other one acts as a full-range retroreflector (FRRR). The signal from this FRRR carries the overall features of local phase modulation, and it is used as the local oscillator for coherent demodulation. With this setup, stationary targets can be rejected at a 4-kHz high-pass cutoff frequency of the filter that follows the demodulator, compared with 20 kHz for conventional fixed-frequency demodulation. This technique features angle insensitivity and provides flow direction as well by implementing standard in-phase and quadrature detection. Besides the direct directional detection of flow signal intensity, flow speed information can be acquired with postprocessing.

Hemorheology↗
PubMed1986

Viral findings in children under the age of two years with expiratory difficulties.

Viral findings were prospectively studied in lower respiratory tract infections in small children with and without expiratory difficulties. On first admission, a viral aetiology was found in 71 of 127 children (56%). On re-admission, a viral etiology was found in only two of 31 cases (6%). Respiratory syncytial viruses (RSV) were responsible for 71% of the cases with viral diagnoses. A recently-developed method for the direct detection of viral antigens in nasopharyngeal specimens by radio-immunoassay was more sensitive than complement fixation serology, especially in patients aged less than six months. Viral diagnosis was reached using this new method alone in 43% of infections caused by RSV and in 27% of infections caused by other viruses. In children under six months, RSV were found in 89% by direct antigen detection and in 22% by serology. We suggest that direct antigen detection should be used as the primary virological method in small children with lower respiratory tract infections. The aetiological agents were the same in cases with and without expiratory difficulties, RSV being found in about 40% of children in both instances. It is concluded that host factors are critical to the development of expiratory difficulties.

Adenoviruses, Human↗
PubMed2004

Amino acid analysis in mammalian cell culture media containing serum and high glucose concentrations by anion exchange chromatography and integrated pulsed amperometric detection.

The direct separation detection of amino acids by anion exchange chromatography with integrated pulsed amperometric detection was optimized for the analysis of typical mammalian cell culture broth samples. Existing gradient elution conditions were adapted, considering the additions of peptone (2 g/L) and 10 vol% fetal calf serum to the medium as well as changing concentrations of glucose from 5.5 g/L up to complete consumption. Samples had to be analyzed in two dilutions with water (1:33.3 and 1:200) due to the strongly varying amino acid concentrations in the samples as a result of the medium composition and cell metabolism. The method was validated in a linear working range for the most common amino acids (2.5-7.5 and 1.25-3.75 microM for cystine/cysteine with 15 microl injection volume). The relative standard deviation of the method for all amino acids was less than 5%, with detection limits of less than 0.6 microM and quantitation limits of less than 1.6 microM. As an example, data for the amino acid composition of different media used for the production of inactivated influenza vaccines in cell culture are shown.

Amino Acids↗
PubMed2006

Complementary study of differential absorption lidar optimization in direct and heterodyne detections.

A detailed study using both analytical and numerical calculations of direct and heterodyne differential absorption lidar (DIAL) techniques is conducted to complement previous studies. The DIAL measurement errors depend on key experimental parameters, some of which can be adjusted to minimize the statistical error. Accordingly, the pertinent criteria on optical thickness, the number of photons emitted at the on and off wavelengths, are discussed to reduce the relative error on the total column content or range-resolved measurements that rely on either hard target or atmospheric backscatter returns. In direct detection, the optimal optical thickness decreases from 1.3 to 0.8 when the background increases while the on-line-to-off-line optimal energy ratio decreases from 3.6 to 2.7. In heterodyne detection, the minimum error is obtained for an optical thickness of 1.2 and an energy ratio of 4.3.

Journal Article↗
PubMed2002

Microbiological aspects of the diagnosis of Chlamydia trachomatis.

The available diagnostic methods for Chlamydia trachomatis infection comprise serology (indirect detection) and culture, antigen detection and nucleic acid amplification (direct detection). The rationale, applications, advantages and disadvantages of the methods and diagnostic targets are discussed. Compared to conventional methods, nucleic acid amplification tests have increased sensitivity. This allows samples to be taken at home by the patient herself and mailed directly to the laboratory. Public health strategies implying home sampling for asymptomatic men and women result in a lower prevalence and a lower risk of short-term complications in terms of pelvic inflammatory disease (PID). The importance of predictive values and the association with prevalence are highlighted.

Antigens, Bacterial↗
PubMed1997

A one step motion detection circuitry.

A model of motion and direction detection is presented as well as results from a computer simulation of the model. The model is based on cooperative computations which allows an extremely simple architecture. Furthermore, it involves only one logical step in its computations even though Hubel and others have argued that motion detection requires at least two. This is made possible by not including the initial detection of movement. The model simply updates prior detections which is a simpler problem. New movement can be captured by the system by means of spontaneous activity in the movement selective cells. The cost is that some iterations are required to capture new movement. On the other hand, the model handles noisy input very well. The reason for this is that movement is viewed as a stable phenomenon rather than change.

Computer Simulation↗
PubMed2005

Separation and detection of peroxynitrite and its metabolites by capillary electrophoresis with UV detection.

A method for the separation and direct detection of peroxynitrite (ONOO(-)) and two of its degradation products, nitrite (NO(2)(-)) and nitrate (NO(3)(-)), using capillary electrophoresis with ultraviolet detection is described. The separation parameters were optimized and included electrokinetic injection, a run buffer consisting of 25 mM K(2)HPO(4) 7.5 mM DTAB, pH 12, and a field strength of -323 V/cm. A diode array UV detector was employed in these studies as it allowed the determination of all three species simultaneously. Nitrate and nitrite provided the maximum response at 214 nm while peroxynitrite generated the best response at 302 nm. All three species could be detected at 214 nm, while simultaneous detection at 214 and 302 nm positively identified each peak.

Electrophoresis, Capillary↗
PubMed1997

Efficiency of the polymerase chain reaction amplification of the uid gene for detection of Escherichia coli in contaminated water.

Direct detection of Escherichia coli from polluted river water was achieved using polymerase chain reaction (PCR) amplification of the uid gene. Amplification using DNA from environmental samples resulted in non-specific DNA fragments. Specific amplification was achieved through use of the touch-down PCR procedure. Targeting the uidA structural region of the gene gave reproducibly better amplification than targeting the uidR regulatory region. The data demonstrate conditions for optimal specific detection.

Bacteriological Techniques↗