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PubMed1993

A rapid screening method for artemisinin and its congeners using ms/ms: search for new analogues in Artemisia annua.

A rapid screening method based on tandem mass spectrometry (ms/ms) is described for artemisinin-related compounds present in complex matrices. These compounds produce abundant ammonium adducts, [M + NH4]+, using ammonia desorption chemical ionization (dci), and dissociation of the mass-selected adducts yields the protonated molecules, [M + H]+, which subsequently eliminate characteristic neutral molecules (H2O, CO, HCO2H, HOAc). Neutral loss ms/ms scans which are selective for different elimination reactions were used in order to screen for groups of related analogues present in a crude hexane extract of Artemisia annua. Comparison of ms/ms product spectra of known Artemisia compounds with those of the new analogues provided information on the functional groups and the molecular weights of the new compounds present in the plant, and tentative structures are suggested.

Antimalarials↗

PubMed2005

Test-retest reliability and practice effects of a rapid screen of mild traumatic brain injury.

Test-retest reliabilities and practice effects of measures from the Rapid Screen of Concussion (RSC), in addition to the Digit Symbol Substitution Test (Digit Symbol), were examined. Twenty five male participants were tested three times; each testing session scheduled a week apart. The test-retest reliability estimates for most measures were reasonably good, ranging from .79 to .97. An exception was the delayed word recall test, which has had a reliability estimate of .66 for the first retest, and .59 for the second retest. Practice effects were evident from Times 1 to 2 on the sentence comprehension and delayed recall subtests of the RSC, Digit Symbol and a composite score. There was also a practice effect of the same magnitude found from Time 2 to Time 3 on Digit Symbol, delayed recall and the composite score. Statistics on measures for both the first and second retest intervals, with associated practice effects, are presented to enable the calculation of reliable change indices (RCI). The RCI may be used to assess any improvement in cognitive functioning after mild Traumatic Brain Injury.

Adolescent↗

PubMed1981

Rapid screening for bacteriuria using a particle counter, pulse-height analyser, and computer.

A system for the rapid screening of urines for the presence of bacteriuria has been devised using a Coulter Counter Model ZBI linked to a multichannel pulse-height analyser (Coulter "Channelyser") with computer analysis of the output. In a series of 215 urines containing growth of a single pathogen of more than 100 x 10(6)/l (greater than 100 000/ml) satisfactory level of sensitivity (99.1%) was obtained using only two different amplification settings by means of a brief treatment (5-10 seconds) of the undiluted specimen with low intensity ultrasound; 85-90% of mixed growths of 10 x 10(6)/l (greater than 10 000/ml) were detected. Sonication did not improve the results in this group. Specimens showing abnormal cyturia of more than 10 x 10(6)/l (greater than 10 000/ml) but no growth on culture were positive in 33% of cases without the use of ultrasound but in 72% after sonication.

Bacteriological Techniques↗

PubMed2003

Rapid screening for Streptococcus agalactiae in vaginal specimens of pregnant women by fluorescent in situ hybridization.

Group B streptococci (GBS) are the most frequent pathogens in neonates with sepsis. A rapid screening method is required to identify carriage of GBS in pregnant women at the time of delivery. In order to detect GBS in vaginal specimens, the efficiency of the standard culture versus fluorescent in situ hybridization (FISH) was investigated. In 258 examined vaginal specimens, FISH identified 58 of the 59 GBS-positive samples (98.3%), whereas by means of standard culture only 38 specimens were positive (64.4%). We recommend FISH as a rapid, specific, highly sensitive screening technique for the detection of GBS in pregnant women at delivery.

Base Sequence↗

PubMed1996

The comparison of a rapid screening MR protocol with a conventional MR protocol for lumbar spondylosis.

PURPOSE: To assess the degree of agreement between a rapid T2-weighted fast spin-echo protocol and a detailed protocol for the detection of lumbar spondylosis and to determine the sensitivity and specificity of the rapid protocol with respect to the detailed protocol. MATERIALS AND METHODS: Forty-eight patients with low back pain were studied with both a detailed four-sequence protocol and a two-sequence rapid screening protocol (acquisition time of 28 and 2.5 min for the detailed and screening protocols, respectively). The two protocol groups were numerically graded for the findings of lumbar spondylosis. RESULTS: Agreement for presence or absence of disc abnormality was 87% for the two techniques. The screening protocol showed a disc abnormality in 91% of all disc herniations and 84% of all disc bulges seen on the detailed study. All moderate and severe (grades two and three in this protocol) bulges and herniations were detected by the screening protocol. Foraminal narrowing, annular tear of the disc, degenerative changes of the facet joints, and nerve root swelling were all better shown on the detailed protocol. CONCLUSION: A rapid, two-plane, single-echo fast spin-echo sequence protocol is adequate to detect potentially significant degenerative disease of the lumbar spine. Its 2.5-min acquisition time allows a complete patient study (including patient preparation) to be performed in less than 10-15 min.

Humans↗

PubMed1994

Rapid screening of the antimicrobial activity of extracts and natural products.

A spectrophotometric method has been developed for the rapid measurement of the antimicrobial activity of natural products, including crude extracts or pure materials. The assay depends on the measurement of non-specific esterase activity using fluorescein diacetate (FDA) hydrolysis in broth cultures of microbes after they have been treated with test compounds. The assay is accurate, reproducible and economical in both time and materials. The speed and economy of the method make it suitable for the rapid screening of many samples and the bioassay directed purification of antimicrobial substances. The assay can also be used with a wide variety of micro-organisms since most micro-organisms are FDA positive. Applications are described in the fields of marine natural products chemistry and essential oils research.

Fluoresceins↗

PubMedDate not supplied

Rapid screening of pharmaceuticals by thin-layer chromatography: analysis of essential drugs by visual methods.

A method for rapidly screening pharmaceuticals by thin-layer chromatography has been designed for use in areas with limited resources and by operators with limited training. An apparatus for performing the analysis in a plastic bag under equilibrium conditions was designed. Results can be reproduced by different operators and in different locations. The analysis can be performed without electricity or in a remote area, away from a laboratory. It is especially suited for field use in developing countries. The method is low cost, maintenance-free, fast, and reliable; it also uses limited volumes of solvents. The analyses can be performed without weighing if reference materials can be supplied in tablet form, provided the drug content is listed and only one unit is required for each analysis. All procedures were developed for the analysis of drugs from a partial list of essential drugs established by the World Health Organization. Three drugs were selected and prepared in the form of reference tablets. Comparisons with the analyses of the drugs in standard dosage forms were made by using reference tablets and primary USP standards. Comparable results were obtained, proving that the screening process can be conducted by using reference tablets and without weighing either the sample or the reference. The method has been successfully demonstrated and used in Swaziland, by high school teachers in the United States, and by personnel from the Ministry of Health in Saudi Arabia. Personnel can be trained in a short time to perform screening analysis of drugs.

Capsules↗

PubMed2003

Rapid screening of binding constants by calibrated competitive 1H NMR spectroscopy.

A calibrated competitive NMR method has been developed that is appropriate for the rapid screening of binding constants. This method involves the initial characterisation of a receptor-substrate binding event for which the (1)H NMR spectrum of a given receptor (calibrant) is modified by the substrate of interest at a range of concentrations. For all subsequent "unknown" receptors, K(a) values are then determined by using a competition assay (in the presence of the calibrant receptor) by measuring a single standard (1)H NMR spectrum. This enables a rapid assessment of the recognition properties of a library of potential receptors. Only the calibrant receptor needs to be NMR active, while the library of putative receptors, as well as the substrate, can be NMR silent. This method assumes the formation of complexes of 1:1 stoichiometry. To demonstrate this methodology, the binding of a number of crown ether type compounds with K+ ions has been studied. Comparison of the binding strengths obtained by using this approach with those in the literature shows excellent agreement. A range of new compounds that have recently been synthesised within our group has also been screened in order to illustrate how this approach can rapidly assess binding ability. This method has significance for chemists working in the fields of combinatorial receptor/substrate design and supramolecular chemistry as a means of rapid optimisation of binding strength.

Journal Article↗

PubMed1995

Rapid screening for resistance to ganciclovir and foscarnet of primary isolates of human cytomegalovirus from culture-positive blood samples.

A rapid screening assay for the detection of resistance to ganciclovir and foscarnet of primary isolates of human cytomegalovirus from culture-positive blood samples was developed by using single doses of both drugs and an immediate-early antigen plaque reduction assay. Results of the rapid assay with peripheral blood leukocytes as the viral inoculum were compared with those of a conventional assay with cell-free virus from the relevant viral isolates. Both assays gave overlapping results. The rapid assay offers the following major advantages: (i) it provides results within 4 to 6 days, (ii) it can be performed with peripheral blood leukocytes, and (iii) it reliably detects drug-resistant human cytomegalovirus strains.

Antigens, Viral↗

PubMed1984

A rapid screening assay for detecting individual RNA species in field isolates of rotaviruses.

A method is described that allows the rapid screening of field isolates of rotavirus for the detection of specific viral RNA segments. Cloned cDNA copies of viral genomic RNAs are employed for detection in the assay which makes use of the dot-hybridization technique of Thomas (1980). The assay developed was shown to be sufficiently sensitive and specific to allow the genetic composition of virions to be determined over a wide range of concentrations. The feasibility of using the method developed for both the analysis of genetic reassortants prepared in vitro and for screening field isolates to detect putative genome reassortment events is demonstrated.

DNA, Recombinant↗

PubMed1996

A rapid screening procedure to identify mini-Tn10 insertion mutants of Escherichia coli K-12 with altered adhesion properties.

A rapid screening procedure was developed for detection of Escherichia coli mutants with altered adhesion abilities using polystyrene 96-well microtiter plates as attachment surfaces. During this assay, bacterial strains grew and adhered simultaneously, and attached cells were measured after crystal violet staining. Starting with a total of 7000 W3110::Tn10 insertion mutants of E. coli K-12 W3110, 50 adhesion-deficient mutants were isolated which showed less than 40% attachment, and 22 mutants were found with an attachment of 40-75%. Motility assays were performed on these 72 mutants, and 34 displayed altered motility.

Bacterial Adhesion↗

PubMed1980

Counter immunoelectrophoresis as a rapid screening test for amoebic liver abscess.

Counter immunoelectrophoresis using cellulose acetate as the supporting medium was used as a rapid screening test for amoebic abscess. All the sera from 40 cases gave positive results. No false positives were obtained, but the results in intestinal amoebiasis were less reliable. An attempt was made to account for discrepancies in previous reports.

Counterimmunoelectrophoresis↗

PubMed1982

Clinical and laboratory efficiency of rapid screening urine cultures by nephelometry.

Urine cultures represent 40-50% of specimens processed by hospital clinical microbiology laboratories. An accurate rapid screening method to detect positive urine cultures could greatly improve laboratory efficiency. Three hundred eighty-two unselected urines were screened by nephelometry at 2, 4, and 5 hours of shaking and incubation in Autobac cuvettes. Results were compared in parallel with standard quantitative-loop plate cultures of the same specimens. Fifty-eight of the 382 specimens showed significant positive (greater than or equal to 10(5) CFU/ml) standard plate cultures. Fifty-four of the 58 were positive at 4 hours by the nephelometric method (sensitivity 93%, 7% false negatives). Plate cultures of the four false negative specimens grew greater than 10(5) CFU/ml Candida (two specimens), Staphylococcus epidermidis or diphtheroids (one specimen each) and represented colonization or contamination without evidence of infection. For 324 specimens negative by plate culture, nephelometry was negative in 287 (89% specificity or 11% false positives). Overall predictive value of a negative test was 99%. Results were comparable for clean-voided or catheterized specimens. The rapid nephelometric urine culture method gave significant savings of material and time.

Catheterization↗

PubMed1998

Rapid screening and comparison of human microsatellite markers in baboons: allele size is conserved, but allele number is not.

We have developed a rapid screening method for cross-species amplification of homologous microsatellite loci and studied some of the characteristics of those loci in the species of origin (human) and the test species (baboon). We tested a matrix of 64 combinations of magnesium concentration and annealing temperature, and after screening over 750 markers with that system, we found that 32 of those combinations account for >99% of the loci that could be amplified across species. This number of test conditions can be further reduced to 6 conditions when a touchdown PCR profile is used instead of traditional 3-step PCR. When the test primers are used to amplify batched DNA samples, and fluorescently labeled dNTPs are incorporated into the PCR product, the products from the best set of conditions can be loaded directly onto an automated sequencer, and the number of alleles, allele size range, and approximate allele frequencies can be estimated from one lane per locus. We were able to optimize conditions for automated microsatellite genotyping in baboons for 24.3% of the loci screened, resulting in a panel of over 280 genetic markers suitable for genetic analysis of baboons. Analysis of 139 markers for which we had completed human and baboon genotyping indicated that there was no correlation between numbers of alleles in humans and in baboons, but a high correlation between median allele sizes. There is no indication of directional selection for increased human microsatellite size.

Alleles↗

PubMed1997

A rapid screening procedure for the identification of high-titer retrovirus packaging clones.

We have developed a viral RNA (vRNA) dot blot assay for rapid identification of high-titer retrovirus vector production by packaging cell clones. The procedure employs Trizol LS reagent to purify vRNA from packaging cell supernatants, a sensitive dot blot assay, and Phosphorlmager technology to quantify packaged viral genomes in 2 days. Experiments performed on viral supernatants of known biological titer demonstrated that the vRNA dot blot assay was extremely sensitive and that dot intensity correlated directly with viral titer. It is often necessary to analyze approximately 100 virus producing cell clones, making this method useful as a rapid screen to identify the highest virus producing clones. The vRNA dot blot assay consistently identified a subset of candidate high-titer producer cell clones. In three independent screens the supernatant with the highest biological titer was produced by one of the previously defined candidate high-titer producer clones. Our procedure greatly facilitates virus titration by: (1) rapidly eliminating the vast majority of low-titer producer cell clones; (2) accurately identifying the subset of candidate high-titer producer clones for further biological titration and assessment of the proviral genomic structure; and (3) reducing laborious tissue culture manipulations to a minimum. Furthermore, the reliance of this method on molecular detection makes it ideally suited for the isolation of high-titer clones lacking a drug selection marker.

Gene Transfer Techniques↗

PubMed1985

Rapid screening of monoclonal antibodies by spin adherence double immunosorbent test (SADIST).

The spin adherence double immunosorbent test (SADIST) is a simple, rapid immunoassay with sensitivity similar to the enzyme-linked immunosorbent assay (ELISA). A 1-step SADIST has been found suitable for rapid screening of hybridomas for antigen-specific monoclonal antibodies (MAb). In this procedure hybridoma supernatants are added to antigen coated microplates followed by commercially available antiglobulin beads. The microplate is immediately centrifuged. Wells containing antigen-specific MAb produce a mat of beads whilst wells without antigen-specific MAb produce a button of beads. No washing or incubation steps are necessary and results are read within minutes of adding beads to test supernatants. By comparison, ELISA tests require several hours to perform with multiple wash steps and further reagent additions. A 2-step SADIST was also assessed. Supernatants are incubated in the microplate as for an ELISA and a wash step precedes the addition of antiglobulin beads. A panel of 117 hybridoma supernatants was selected to assess the suitability of the SADIST techniques for hybridoma screening. The supernatants were added to antigen-coated microplates and SADIST and ELISA tests performed. The SADIST correctly discriminated most hybridoma supernatants that were clearly positive or negative by ELISA. It was also found possible to perform SADIST followed by ELISA tests on the same microplate well without significantly affecting ELISA values.

Antibodies, Monoclonal↗

PubMed2003

A rapid screening assay for inhibitors of 11beta-hydroxysteroid dehydrogenases (11beta-HSD): flavanone selectively inhibits 11beta-HSD1 reductase activity.

A rapid screening assay for chemicals inhibiting 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1 or type 2 using lysates from stably transfected cells was developed. Here, we tested a series of environmental chemicals for anti-11beta-HSD activities. Inhibition of 11beta-HSD2, which may cause cortisol-dependent activation of the mineralocorticoid receptor with sodium retention and hypertension, was observed for several compounds, with diethylcarbamate being the most potent inhibitor (IC50 6.3 microM). Abietic acid inhibited both 11beta-HSD1 (IC50 27 microM for reduction and 2.8 microM for oxidation) and 11beta-HSD2 (IC50 12 microM). Our results demonstrate for the first time that flavanone selectively inhibits 11beta-HSD1 reductase activity: this enzyme being considered as essential for the local activation of glucocorticoids and representing a potential target for the therapeutic treatment of diabetes type 2. Flavanone and 2'-hydroxyflavanone efficiently inhibited reductive (IC50 18 and 10 microM) but not oxidative activity. We observed a reduced inhibitory effect of hydroxylated flavanone derivatives and of flavones containing a double-bond between atom C2 and C3. Flavanone was specific for 11beta-HSD1 and did not inhibit 11beta-HSD2. Our results reveal that a variety of environmental compounds exert distinct inhibitory effects on 11beta-HSD1 and 11beta-HSD2, opening the possibility for selectively modulating local cortisone/cortisol availability in vivo.

11-beta-Hydroxysteroid Dehydrogenase Type 1↗

PubMed1997

A molecular strategy designed for the rapid screening of gene traps based on sequence identity and gene expression pattern in adult mice.

We have devised a strategy to rapidly screen gene traps in mouse embryonic stem (ES) cells based on DNA sequence information and an in vitro analysis of gene expression. After the initial identification of ES cell clones expressing beta-galactosidase, tagged RNA transcripts were immediately cloned and sequenced in order to determine their identities. Novel gene sequences found were used to probe northern blots to examine the expression patterns of their cognate genes. Our initial characterization of 30 cDNA clones indicated that more than half of the tagged sequences were novel mouse genes and of these 40% showed a restricted pattern of expression in adult mouse tissues. This molecular characterization of gene traps is quick, reliable and well suited for the large-scale screening of mammalian developmental genes. Furthermore, since gene trap insertion frequently disrupts the tagged host gene, the ES cells can be used to produce transgenic animals for a genetic analysis of gene function.

Animals↗