PubMed HealthSearch

Biomedical subjects

F M Olthuis

Publications and source records attributed to F M Olthuis.

7 recordsLinked to original sources

Determination of the mean normal prothrombin time for assessment of international normalized ratios. Usefulness of lyophilized plasma.

To report a Prothrombin Time (PT) as International Normalized Ratio in controlling oral anticoagulant therapy, the Mean Normal PT (MNPT) is required. To correct for methodological differences in performing the PT test, each laboratory should determine its own MNPT for each batch of reagent using fresh blood samples from a large number of normal individuals. This would be a laborious procedure. Two models for simplified assessment of MNPT were investigated by two laboratories in a collaborative study. According to the models, the MNPT of a new batch of reagent is calculated, using the PT of a lyophilized control plasma measured with the new batch and a reference batch, as well as the MNPT of the reference batch obtained with fresh samples. Experimental results were obtained with 19 batches of bovine thromboplastin, 4 lyophilized normal control plasmas and fresh blood samples of 40 normal individuals. The PTs of the 4 lyophilized normal control plasmas were not identical to the MNPT of the fresh normal samples and also different from each other. Therefore, the uncorrected PTs of these control plasmas cannot be used as MNPT. In general, there was good agreement between measured and calculated MNPT, although some control plasmas gave better results than others. There were no significant differences between the results obtained by both calculation models.

Freeze Drying

A Multi-centre study to evaluate method dependency of the international sensitivity index of bovine thromboplastin.

In The Netherlands, a particular coagulometer method for prothrombin time (PT) determination with reduced sample and reagent volumes is used by 62% of the laboratories controlling oral anticoagulant therapy. This "micro-method" has been calibrated against the manual tilt-tube technique for PT determination by six Dutch laboratories. Each laboratory tested 20 fresh normal blood samples and 60 fresh patient blood samples using both methods with the same batch of bovine thromboplastin reagent, according to a detailed protocol. Both methods were comparable as to their precision, but PTs measured by the micro-method were significantly prolonged (p less than 0.001, Student's t-test) as compared to the manual method. This effect is stronger for samples of normal subjects than for patients' samples. It was assumed that the International Sensitivity Index (ISI) of the bovine thromboplastin for the manual method was 1.00 in each laboratory. The ISI-values of the bovine thromboplastin for the micro-method determined by the six laboratories ranged from 1.00 to 1.07 (mean 1.03, SD 0.03). Our results indicate that any other laboratory, using this thromboplastin and the micro-method, should obtain accurate assessment of the International Normalized Ratio from their own mean normal PT and an ISI which is 3% higher than the ISI supplied by the thromboplastin manufacturer for the manual tilt-tube method.

Animals

The effect of radiolabeling of human fibrinogen on its adsorption behaviour on a polystyrene surface.

Human fibrinogen (HFB) was labeled with different radioactive labels (Technetium -99m and iodine -125) in various ways. Characterization by chromatographic and electrophoretic methods did not show differences between the labeled and the nonlabeled proteins. The effect of the label and the labeling method on the adsorption behaviour of 99mTc and 125I labeled HFB at a polystyrene surface was investigated. In all cases labeled HFB showed preferential adsorption as compared to nonlabeled HFB. The preferential adsorption was expressed in terms of a factor ø (van der Scheer et al. 1978a), which will be 1, when no preferential adsorption occurs. 99mTc - and 125I - HFB showed ø values from 1.48 - 1.88. It is concluded that only meaningful adsorption experiments with labeled proteins can be performed when the possible occurrence of preferential adsorption has been investigated by appropriate methods. The results of prior work on protein adsorption at biomaterials using radiolabeled proteins have to be reconsidered.

Adsorption

Urinary oxalate estimation.

A method of estimating urinary oxalate is described. Impurities which might otherwise interfere with the estimation are removed by means of an ion-exchange resin. The oxalate is then reduced to glycolic acid, which forms a coloured compound with chromotropic acid. It can then be estimated colorimetrically. No special equipment is required, oxalate recovery is good, and the method is reproducible and precise.

Chromatography, Ion Exchange

Oxalic acid determination in plasma.

A method is described for the estimation of oxalic acid in plasma. After removing interfering substances with an ion-exchange resin, oxalic acid is reduced to glycolic acid, which forms a coloured compound with chromotropic acid. No special equipment is required. A good recovery and precision are found.

Chromatography, Ion Exchange