Fluorescence polarization immunoassay of gentamicin or netilmicin in blood spotted on filter paper.
(25/32)In this improved simple method for determination of aminoglycoside antibiotics in dried-blood spots on filter paper, gentamicin or netilmicin is recovered from the blood spot most effectively by incubation for 60 min in an ultrafiltration tube containing 500 microL of 0.5 mol/L Na2HPO4 buffer. The eluates from the paper are centrifuged, then transferred to an Abbott TDx cartridge for measurement of gentamicin or netilmicin by fluorescence polarization immunoassay. The dried sample on paper is stable for about eight days at ambient temperature. Intra-assay CVs for gentamicin and netilmicin are less than 8.5% and less than 6.1%, respectively. Analytical recovery of gentamicin and netilmicin from the paper exceeded 90%. This method permits simple blood collection and monitoring of the therapeutic concentration of gentamicin or netilmicin in serum, particularly that of newborn infants and small children. (+info)
Acute and chronic pharmacokinetics of asymmetrical doses of slow release choline theophyllinate in asthma.
(26/32)The day and night pharmacokinetics of assymetrical doses of slow release choline theophyllinate (Sabidal SR 270) were compared at day 1 and at day 4 of treatment when steady state had been achieved. Ten patients with chronic asthma were given oral choline theophyllinate 424 mg at 09.00 h and 848 mg at 21.00 h for 4 days. At regular intervals during day 1 and day 4 of treatment theophylline concentrations were measured in plasma and dried blood spots by fluorimmunoassay. Theophylline concentrations measured from dried blood spots were slightly lower than those in plasma, the difference remaining constant at all time points during day 1 and day 4 of treatment. On day 1 the mean peak plasma theophylline concentration was 5.4 +/- 1.0 (+/- s.e. mean) micrograms ml-1 4 h after the morning dose and 11.2 +/- 1.6 micrograms ml-1 4 h after the evening dose which were significantly (P less than 0.01) different. Similarly the areas under the plasma theophylline concentration-time curves at night were significantly (P less than 0.001) greater than those observed during the day. During day 4 mean peak plasma concentrations of theophylline after the morning and larger evening dose were 13.2 +/- 1.3 and 12.1 +/- 1.4 micrograms ml-1 respectively, which were not significantly different. No significant difference was observed between the areas under the plasma theophylline concentration-time curves during the day and at night. However the post-dose time to peak was significantly delayed at night (6 h) compared to the morning (2 h, P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) (+info)
Identification from crossed electro-immunodiffusion patterns of serum.
(27/32)An attempt to identify the individuality from crossed electro-immunodiffusion (CEID) patterns of serum was made by using coefficient of variation (CV), the ratio of standard deviation to mean of ratios of area in couples of corresponding peaks in the CEID patterns. The values of CV in most cases of sera from identical subject were less than 10 except only one case of 10.6. On the contrary, those in most cases of sera from different subjects were more than 20 except a few cases of 14.9 to 20. No overlap of CV values was observed between the two groups. No significant differences were demonstrated in three groups of sex pairing, i.e., male-male, female-female and female-male. From these results, analysis of CEID patterns by employing CV was found to be available for fairly definite identification as well as differentiation but useless for distinction of sexes in cases of different subjects. (+info)
Immunoelectrophoretic analyses of antigenic and human specific proteins of red cell membrane.
(28/32)Rabbit antisera were raised for red cell stroma extracted with each of the following solutions; 5 mM phosphate buffer pH 8.0 (A antigen). 5 mM EDTA containing 5 mM 2-mercaptoethanol (B), 1/15 M phosphate buffered saline (C), 0.05% SDS (D), 50 mM Tris-HCl buffer pH 8.0 containing 1% SDS, 1 mM EDTA and 1% 2-mercaptoethanol (E) and 1% Triton X-100 (F). Crossed immunoelectrophoresis showed that E antigen produced relatively numerous precipitation lines, 7-9, against each of the antisera, suggesting that E solution was superior to the others for membrane solubilization. However, the Ouchterlony test demonstrated potent precipitin activity in anti-C, and the antiserum absorbed with monkey stroma which was extracted with F solution was proved to have strict human specificity. By adopting the antiserum to affinity chromatography, human specific antigens were isolated. The antigens gave four bands moving faster than Band 5 at SDS-polyacrylamide gel electrophoresis none of which was stained by Schiff's reagent. Using absorbed anti-C, human specificity of blood stains kept us to for two years could be identified. (+info)
A simple method to detect AB antigen in blood grouping of body fluid stains.
(29/32)A modified mixed agglutination method is described for detecting AB antigen from body fluid stain. Saline extract of body fluid stain was added to A.Se saliva stain which was beforehand sensitized with anti-A. AB antigen reacting with the stain was detected with the usual mixed agglutination method using anti-B and B red cells. (+info)
Fragmentation of human albumin with proteolytic enzymes and its antigenicity with special reference to human-specificity.
(30/32)Human albumin prepared by the trichloroacetic acid method was treated with bromelin, ficin, papain, pronase or trypsin. Pronase- or trypsin-treated albumin showed four fragments on polyacrylamide disc electrophoresis. When rabbit antiserum to pronase-treated albumin was adsorbed with monkey albumin, antibody with human specificity was completely abolished, whereas anti-albumin serum adsorbed with pronase-treated albumin retained the antibody to human albumin. It was considered that antigenic site with human specificity in albumin structure was inactivated by pronase. (+info)
Preparation of anti-alpha 2-macroglobulin using Canavalia lineata DC lectin for differentiating species-specificity of blood stains.
(31/32)To differentiate species-specificity of blood stains, anti-alpha 2-macroglobulin was raised in rabbits against Canavalia lineata DC lectin-serum complex (LSC). Adsorption of anti-LSC with human lipoprotein resulted in antiserum specific for alpha 2-macroglobulin. It was confirmed by Ouchterlony test that the antiserum adsorbed successively with monkey serum or anti-LSC adsorbed directly with monkey serum reacted with only human serum but not with mammalian ones. Immunoelectrosyneresis and anti-LSC consumption test could identify species-specificity of blood stains kept for up to two years and for up to several years, respectively. It is indicated that anti-LSC is quite effective for differentiating species-specificity of blood stains. (+info)
The analysis of EDTA in dried bloodstains by electrospray LC-MS-MS and ion chromatography.
(32/32)Analytical methods were developed to determine the presence of ethylenediaminetetraacetic acid (EDTA) in dried bloodstains to provide probative information when allegations of evidence tampering have been made in criminal cases. A simple screening method using ion chromatography to analyze stains was found to be quantitative to the 5 ppm level. The presence of EDTA was then confirmed using negative and positive ion mode liquid chromatography-tandem mass spectrometry (LC-MS-MS) methods. A blind trial of these methods on 42 samples correctly determined the bloodstains that did and did not contain the preservative EDTA. One interesting observation in these results was the adsorption and postanalysis release of EDTA in the chromatographic system. In order to avoid cross contamination of samples resulting from this phenomena, it was found to be necessary to use EDTA-free blood extracts as blanks in the LC-MS analysis of bloodstains. (+info)