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A rapid screening test for Cushing's syndrome.

A combination test consisting of two parts, a single dose of dexamethasone and a (1-24) ACTH injection, has been used for the diagnosis of Cushing's syndrome. The test was performed by administering 1 mg dexamethasone orally at 11 p.m. on the first day and 25 IU synthetic ACTH i.v. at 8 a.m. on the second day. Plasma cortisol was determined at 8 a.m. on the first and second day and 1-1/2 and 2 hours after the ACTH injection. The test was performed in 33 patients with Cushing's syndrome and in 114 controls without adrenal disease. After dexamethasone administration alone, cortisol values frequently overlapped between patients with Cushing's syndrome and controls. The validity of the test was considerably improved by adding ACTH stimulation. The test procedure is recommended as a screening test for Cushing's syndrome in ambulatory practice.

Adrenocorticotropic Hormone

Rapid screening and identification of genes involved in bacterial extracellular membrane vesicle production using a curvature-sensing peptide.

Bacteria secrete extracellular membrane vesicles (EMVs). Physiological functions and biotechnological applications of these lipid nanoparticles have been attracting significant attention. However, the details of the molecular basis of EMV biogenesis have not yet been fully elucidated. In our previous work, an N-terminus-substituted FAAV peptide labeled with nitrobenzoxadiazole (NBD; nFAAV5-NBD) was developed. This peptide can sense the curvature of a lipid bilayer and selectively bind to EMVs even in the presence of cells. Here, we applied nFAAV5-NBD to a genome-wide screening of hyper- and hypo-vesiculation transposon mutants of a Gram-negative bacterium, Shewanella vesiculosa HM13, to identify the genes involved in EMV production. We analyzed the transposon insertion sites in hyper- and hypo-vesiculation mutants and identified 16 and six genes, respectively, with a transposon inserted within or near them. Targeted gene-disrupted mutants of the identified genes showed that the lack of putative dipeptidyl carboxypeptidase, glutamate synthase β-subunit, LapG protease, metallohydrolase, RNA polymerase sigma-54 factor, inactive transglutaminase, PepSY domain-containing protein, and Rhs-family protein caused EMV overproduction. On the other hand, disruption of the genes encoding putative phosphoenolpyruvate synthase, d-hexose-6-phosphate epimerase, NAD-specific glutamate dehydrogenase, and sensory box histidine kinase/response regulator decreased EMV production. This study demonstrates the utility of a novel screening method using a curvature-sensing peptide for mutants with altered EMV productivity and provides information on the genes related to EMV production.IMPORTANCEConventional methods for isolation and quantification of extracellular membrane vesicles (EMVs) are generally time-consuming. nFAAV5-NBD can detect EMVs in the culture without separating EMVs from cells. In situ detection of EMVs using this peptide facilitated screening of the genes related to EMV production. We succeeded in identifying various genes associated with EMV production of Shewanella vesiculosa HM13, which would contribute to the elucidation of bacterial EMV formation mechanisms. Additionally, the hyper-vesiculating mutants obtained in this study would be valuable for EMV applications, such as secreting useful substances as EMV cargoes and producing artificially functionalized EMVs.

Shewanella

Rapid screening for significant bacteriuria using a Coulter Counter.

A Coulter Counter was evaluated for detecting significant bacteriuria. Simple urinary particle counts showed agreement in 883 of the 956 (92.4%) urine specimens examined compared to a standard culture procedure. They also provided a semiquantitative estimate of bacterial numbers in specimens diagnosed as infected by culture. Sample preparation before counting was minimal, and results were obtained within two to four minutes.

Bacteriological Techniques

A rapid screening test for reduced fibrinolytic activity of plasma: streptokinase activated lysis time.

A simple screening method for determining the fibrinolytic activity of plasma is described. A streptokinase activated system is used, which measures the result of the interplay of all components of the fibrinolytic system with the exception of activators, which are added in excess. Mean lysis time and standard deviation with this method is 6.9 +/- 1.4 min. The sensitivity of the method in detecting minor delays in clot lysis time is demonstrated.

Blood Coagulation Tests

[The differentiation of clostridial microcultures on nuclepore-filters by fluorescent antibodies as a rapid screening test for spores. I. Method of producing microcultures (author's transl)].

Spores of 32 strains of 11 different clostridial species (Table 1) were allowed to germinate as microcultures on Nuclepore membrane-filters to ascertain whether they could be differentiated according to their patterns of growth. The questions dealt with in the preliminary studies, investigated which pore-size of filters would be best for this purpose and which time necessary for each species to produce microcolonies of 10 to 20 cells from a single spore. Nuclepore-filters with a pore-diameter of 1 mu gave the best results and the times for producing microcolonies of the desired size ranged from 4 hours with Cl. septicum up to 10 hours with Cl. tetani (Table 3). The final result of the study was, that the differences in the patterns of growth were not great enough to allow identification. Only Cl. perfringens formed specific and distinctive compact colonies (Tables 4 and 5).

Bacteriological Techniques

A rapid screening for the specific DNA sequence: analysis of transforming DNA segments in adenovirus-transformed cells.

The viral DNA sequences in cells transformed by adenovirus type-12 (Ad12) DNA fragments were investigated by spot hybridization, a detection by autoradiography of nucleic acid hybrids formed between cell DNAs spotted on a membrane filter and various nick-translated Ad12 DNA fragments. In CY1 cell line, a rat cell line transformed by the EcoRI-C fragment (left hand 16%), all of the HindIII fragments included in the EcoRI-C fragment are shown to be present. In GY1 cell line, a rat cell line transformed by the HindIII-G fragment (left hand 7%), both of the BpaI-H and a part of the BpaI-J, two components consisting of the HindIII-G fragment, were found. A dominant presence of the Ad12 BpaI-H fragment (left hand end 4.5%) of the Ad12 DNA molecule, approximately 60% of the Ad12 transforming DNA sequences (Ad12 HindIII-G), was also shown in GY1. Spot hybridization used in the experiment will be of general use for detecting viral nucleic acid sequences in cells and provides a simple and useful screening method for investigating viral etiology of tumors and transformed cells.

Adenoviruses, Human

Rapid screening for chemical carcinogens: transforming activity of selected nitroso compounds detected in a transplacental host-mediated culture system.

The transplacental host-mediated hamster cell culture system was used to test a variety of solvents and chemicals of unknown and known (positive and negative) activity for their ability to induce morphologic transformation of cells and growth in agar. Examination of approximately 13,000 colonies of cells from untreated animals yielded no transformants, thus demonstrating no spontaneous transformation in the system. Similar negative results were obtained after animals were treated with the solvents acetone, ethanol, dimethyl sulfoxide, dimethylformamide, and trioctanoin oil. Several known carcinogens, including benzo[a]pyrene, methylnitrosourethane, urethan, and diethylnitrosamine, were positive for transforming activity. Three pesticides, carbaryl, methomyl, and landrin, and their N-nitroso derivatives were tested. All the nitrosated forms had transforming activity, but only one of the pesticides, landrin, was positive. In all experiments conducted, results of the agar-growth test correlated well with tests for morphologic transformation. The transplacental hamster embryol cell culture system therefore detected transforming activity of N-nitroso compounds and some known carcinogens.

Agar

Rapid screening method for alkaline phosphatase activity in cheese: collaborative study.

The method developed for developed for determining alkaline phosphatase activity in cheese, in which phenolphthalein monophosphate is used as the substrate, was collaboratively studied. A 7.5% butanol extract of cheese is reacted with phenolphthalein monophosphate; phenolphthalein is released and yields a red solution that is compared visually with a standard (s) prepared from the same extract. Seven collaborators analyzed 8 samples of cheese, in duplicate, by the screening method and Scharer I method. Of the 208 observations returned, only 4 were incorrect. The alkaline phosphatase method has been adopted as official first action.

Alkaline Phosphatase