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At least 865 records · Page 48Linked to original sources

Altered isozyme patterns of leucocyte alkaline phosphatase in disease states.

Leucocyte alkaline phosphatase (LAP) is a granulocyte enzyme whose concentration varies in disease states. In order to determine whether the pattern of expression is altered in leukaemic granulocytes, we have analysed the LAP isozyme pattern of a series of normal subjects and patients with various haematological diseases. Electrophoretic patterns of partially purified LAP samples were determined by polyacrylamide gel electrophoresis in the presence of Triton X-100. These patterns were reproducible on repeated samples from the same patient. Presence of the LAPf and LAPs isozymes were determined after staining with the dye Fast Blue BB. Granulocytes were obtained from 15 normal subjects. Thirteen of these samples had only the LAPs isozyme. The other two had LAPs, plus a small amount (less than or equal to 10% of total) of LAPf activity. Eight patients with stable phase chronic myelogenous leukaemia (CML) had only small amounts of the normal LAPs isozyme and no evidence of LAPf . Of 11 patients with CGL who clinically had blast crisis. 10 had both LAPs and LAPf . The eleventh patient who was Ph1 negative had only LAPs. Three of five patients with polycythaemia vera had only the LAPs isozyme while two had both isozymes. Six patients with non-malignant leucocytosis had only LAPs. We interpret this data to indicate that the increased levels of LAP activity in some CGL blast crisis patients are primarily related to synthesis of the LAPf isozyme.

Alkaline Phosphatase↗

[Glucocorticoid receptors in normal human lymphocytes].

Glucocorticoid (GC) receptors were studied in intact lymphocytes from 11 donors. GC binding parameters were found to be highly reproducible in repeated experiments with lymphocytes. It was shown that GC receptors in donors' lymphocytes could be distributed into two different classes similarly to the pattern seen in skin fibroblasts. Human lymphocytes are an adequate object for studying genetically determined variability of GC receptors and its clinical importance.

Adult↗

[Adaptation of IgG, IgA, and IgM determination on the Isamat: a turbidimetric method].

We have adapted the turbidimetric assay of immunoglobulins IgG, IgA and IgM to a transfer analyser: Isamat. This reaction takes place in the presence of PEG and the reading is made at 340 nm. The repeatability and reproducibility are satisfactory with a variation coefficient of 2.9% to 7.2%. The correlation with the kinetic method (ICS Beckman) is excellent for IgG and IgA and good for IgM. The protocol established enables eleven assays of three different proteins in one hour. The reduced requirement for pure antiserum and the low sample volume (5 to 10 microliters) make this turbidimetric technique useful in pediatrics.

Autoanalysis↗

Protein efficiency ratio: AACC/ASTM collaborative study.

Eight laboratories (7 of the laboratories conducted animal experiments) participated in a collaborative study to standardize some of the methodology associated with animal bioassays for determining protein efficiency ratios and to suggest improvements which would reduce the variation among laboratories. One-, 2-, 3-, and 4-week protein efficiency ratios (PER) with 0-, 2-, or 4-day adaptation periods were obtained from each laboratory, respectively, for 6 protein sources: casein, lean beef, lactalbumin, textured vegetable protein, peanut flour, and wheat flour. Analyses were computed for PER and adjusted PER (APER). From the analysis of variance for PER and APER, significant (P less than 0.05) effects were observed due to laboratories, adaptation length, protein sources, and/or interactions among these variables. In general, APER values show much less variation among laboratories than PER values. The reproducibility and repeatability variances were significantly (P less than 0.05) greater for an assay length of 2 weeks than they were for 3- or 4-week assays. Two protein sources, casein and textured vegetable protein, were fed at both high (10%) and low (6%) levels of protein. Analysis of variance of PER values shows a significant (P less than 0.05) laboratory by protein level by assay length interaction.

Animals↗

Determination of apoprotein B in the skin.

To determine the origin of cutaneous cholesterol a method has been developed for extracting and determining by electro-immuno-assay apoprotein B in the skin as already performed by Hoff at the arterial level. This study demonstrated the presence of apoprotein B in the skin and its quantification. The easy and reliable technique employed required only conventional laboratory instruments. Repeatability and reproducibility studies give encouraging results comparable with those obtained on arterial tissue. They show that at least one part of skin cholesterol originates in low density lipoproteins.

Apolipoproteins↗

Determination of N-nitrosodimethylamine in nonfat dry milk: collaborative study.

Ten laboratories participated in a collaborative study of a method for the determination of N-nitrosodimethylamine (NDMA) in nonfat dry milk. NDMA is eluted with dichloromethane from a mixture of Celite, acidic sulfamic acid, and nonfat dry milk (all packed in a chromatography column), concentrated in a Kuderna -Danish concentrator, and finally analyzed by a GC-thermal energy analyzer technique. Ten samples were studied: 6 were naturally contaminated (NDMA levels 0.38-3.56 ppb) and 4 were spiked with known levels (0.96 and 3.2 ppb) of NDMA. The coefficients of variation (CV) of the complete data for the naturally contaminated samples (excluding the 2 samples containing the lowest levels) were 8.5% and 22.5% for repeatability and reproducibility, respectively. The corresponding CVs for the spiked samples were 14.4% and 20.4%, respectively. The percent recoveries of the added NDMA in the spiked samples (at the 2 levels indicated above) were 101.6 +/- 3.2 (omitting 1 outlier) and 95 +/- 2.1, respectively. The method has been adopted official first action.

Animals↗

Dry column-thermal energy analyzer method for determining N-nitrosopyrrolidine in fried bacon: collaborative study.

A dry column method for isolating N-nitrosopyrrolidine (NPYR) from fried, cure-pumped bacon and detection by gas chromatography-thermal energy analyzer (TEA) was studied collaboratively. Testing the results obtained from 11 collaborators for homogeneous variances among samples resulted in splitting the nonzero samples into 2 groups of sample levels, each with similar variances. Outlying results were identified by AOAC-recommended procedures, and laboratories having outliers within a group were excluded. Results from the 9 collaborators remaining in the low group yielded coefficients of variation (CV) of 6.00% and 7.47% for repeatability and reproducibility, respectively, and the 8 collaborators remaining in the high group yielded CV values of 5.64% and 13.72%, respectively. An 85.2% overall average recovery of the N-nitrosoazetidine internal standard was obtained with an average laboratory CV of 10.5%. The method has been adopted official first action as an alternative to the mineral oil distillation-TEA screening procedure.

Animals↗

[Comparative analysis of the virion structural proteins of 3 nuclear polyhedrosis viruses].

The composition of structural proteins of virions of nuclear polyhedrosis viruses (NPV) of Barathra brassicae L., Lymantria dispor L. and Aporia crataegi L. was studied by polyacrylamide gel-SDS electrophoresis (PAGE-SDS). The object of the study included fractions of virion bundles in the envelope. All the baculoviruses under study were shown to have in virion structural proteins 23-24 polypeptides with molecular weights ranging from 12500 to 95000 daltons. Alongside with polypeptides of similar molecular weight, each baculovirus was shown to have a specific set of structural proteins allowing them to be readily identified. Specific distribution of major polypeptides of the virions was found. All the protein fractions were readily reproducible in repeated analyses with the exception of two minor polypeptides of NPV of Barathra brassicae L. The study showed that the composition of structural proteins of virions may be an important criterion for identification of baculoviruses under identical electrophoresis conditions.

Electrophoresis, Polyacrylamide Gel↗

Enumeration and confirmation of Bacillus cereus in foods: collaborative study.

A collaborative study was conducted in 15 laboratories to evaluate 2 different techniques for enumerating Bacillus cereus in foods. A direct plating technique using mannitol-egg yolk-poly-myxin agar and a most probable number (MPN) technique using trypticase-soy-polymyxin broth were compared for the enumeration of high and low populations of B. cereus in mashed potatoes. The collaborative results showed that the overall mean recovery obtained with the low population level was essentially the same by both techniques. However, the overall mean recovery was significantly higher by the direct plating technique at the high population level. A statistical evaluation of the data also showed that the direct plating technique had better repeatability and reproducibility than did the MPN technique at both the high and low population levels. These results suggest that the MPN technique is suitable for examining foods containing low populations of B. cereus, but that the direct plating technique is preferable for foods that contain a high population of this organism. The confirmatory technique used in the proposed method is reliable for presumptive identification of isolates as B. cereus. The method has been adopted as official first action.

Bacillus cereus↗

Analysis of fat-soluble vitamins. XXVIII. High performance liquid chromatographic determination of vitamin D in pet foods and feeds: collaborative study.

A high performance liquid chromatographic (HPLC) method for vitamin D in pet foods and feeds at low concentrations (2-8 IU/g = 50-200 ppb) was studied collaboratively. The procedure consists of the following purification steps: saponification, extraction of the unsaponifiable fraction, chromatography on alumina, cleanup on reverse phase HPLC, and quantitation with straight phase HPLC. The original method, developed by Knapstein, was simplified by deleting the quantitative TLC step. Six coded samples were distributed to 31 laboratories, along with a known sample containing 15 IU/g to allow practice of the rather complicated procedure. Eighteen collaborators returned their results. Results for the spiked samples show good recovery. The estimates of repeatability and reproducibility are 0.96 and 2.2 IU/g for spiked samples and 1.5 and 3.1 IU/g for commercial samples, respectively, which are considered acceptable for these low concentrations. The method has been adopted official first action.

Animal Feed↗

Quantitative thin layer chromatographic multi-sulfonamide screening procedure: collaborative study.

A thin layer chromatographic procedure suitable for detection of multiple sulfonamides at 0.1 ppm was studied in an interlaboratory collaborative study. Sulfamethazine, sulfadimethoxine, and sulfaquinoxaline were variously analyzed in liver and muscle tissues from swine, turkey, and duck. The average recovery for all drugs across all tissues was 95%. The corresponding repeatability and reproducibility were 7.7% and 10.5%, respectively.

Animals↗

[A contribution to stepless antibody determination by the "enzyme linked immunosorbent assay" (ELISA) (author's transl)].

A simple method for the mathematical calculation of ELISA results is described. Using the formula (formula: see text) (Eu, En, Ep = extinction values of the unknown serum, the negative, and positive standard serum, respectively) an index value Iu is calculated for the testserum, whereby the optical density of the testserum is put in relation to the optical density of a negative and positive standardserum. Using three different ELISA test systems, the Iu values of a number of human, bovine and rabbit testsera were relatively uniform and reproducible in repeated investigations. A statistically calculated tolerance limit for the index values of negative sera can be introduced into the formula described, thus facilitating the decision "negative" or "positive" for unknown sera.

Animals↗

Gas-liquid chromatographic-thermal energy analyzer determination of N-nitrosodimethylamine in beer: collaborative study.

The GLC/TEA method for N-nitrosodimethylamine (NDMA) in beer was studied collaboratively by 13 laboratories from 7 countries. Collaborators were asked to analyze a total of 10 randomly labeled samples of beer consisting of the following duplicates: a naturally contaminated commercial beer; a beer extremely low (ca 0.1 ppb) in NDMA; and the low NDMA beer spiked with 0.5, 1.9, and 5.0 ppb NDMA. The pooled repeatability and reproducibility coefficients of variation (CV) for all samples were 17% and 27%, respectively. However, when data from 2 laboratories (outliers) were omitted, the corresponding CV values improved considerably (11% and 15%, respectively). Variance analysis showed the presence of a significant laboratory-sample interaction when all data were used for analysis, but this interaction disappeared when data from the 2 outlying laboratories were excluded. The pooled percent recovery of the overall method (omitting outliers) was 101.4 +/- 3.5. All the laboratories detected NDMA in the low NDMA beer. The method was adopted official first action.

Beer↗

Differential pulse polarographic determination of iodine in thyroid tablets: collaborative study.

A differential pulse polarographic (DPP) method for the determination of iodine in thyroid and thyroid preparations was collaboratively studied by 8 laboratories. The overall concentration of iodine in commercial thyroid tablets containing 1/4, 1,2, and 5 gr. of declared thyroid was 0.196%. The overall repeatability and reproducibility standard deviations were 0.0043 and 0.0067, respectively, and the corresponding coefficients of variation were 2.18 and 3.41%, respectively. The results obtained by the DPP method agreed with those obtained by the U.S. Pharmacopeia XX method at the various concentrations of declared thyroid studied. The DPP method has been adopted official first action.

Iodine↗

Comparison of three methods for determination of crude protein in meat: collaborative study.

A study was designed to compare the Kjel-Foss automated macro-Kjeldahl method and a block digestion-steam distillation method. The official AOAC Kjeldahl method was used as a reference procedure. Six products with a crude protein range of 10-30% were analyzed by 23 laboratories. Five laboratories analyzed the samples by the official AOAC method, 8 laboratories used the automated Kjel-Foss method, and 11 laboratories used the block digestion with steam distillation method. Standard deviations for each product and each method for both repeatability and reproducibility are given. The block digestion-steam distillation method has been adopted official first action.

Animals↗

Dual column chromatographic method for determination of N-nitrosothiazolidine in fried bacon.

A dual column chromatographic procedure is described that permits the determination of N-nitrosothiazolidine (NTHZ), a newly identified nitrosamine, and N-nitrosopyrrolidine (NPYR), the principal volatile nitrosamine, in the same fried bacon sample. The procedure used an acid-Celite, Celite, sodium sulfate column followed by a partially deactivated basic alumina column to isolate NTHZ from bacon for subsequent quantitation by gas chromatography with a thermal energy analyzer (GC-TEA). A limited intralaboratory study gave standard deviations for repeatability and reproducibility of 1.20 and 1.55 ppb, respectively. The recovery of N-nitrosothiomorpholine, the internal standard added at the 10 ppb level, was 93.3 +/- 6.03%. The method is also free from artifactual nitrosamine formation.

Animals↗

[Variability of the heterochromatic regions of human chromosomes 1, 9, 16, and Y].

The variability of heterochromatic regions on chromosomes 1, 9, 16 and Y has been studied in a group of unrelated subjects (50 boys and 50 girls) using quantitative methods. Analysis of intercellular (with one subject) variability of chromosome C-segments lengths has shown that their absolute lengths vary considerably with the cell sample examined, while the relative lengths (a part of the total length of C-segment of the chromosomes being studied) are more stable. The relative lengths may be helpful in individual characteristics, reproducible in repeated analysis, which may be used in zygosity testing in twins, family analysis, etc. Absolute lengths, however, may be successfully used in population studies, as the difference between the cell samples peculiarities on which the C-segment length is based in particular karyotypes, is obliterated. Comparative analysis of mean lengths and variability of C-segments on chromosomes 1, 9, 16 in the groups of boys and girls has revealed no differences between the sexes. The mean lengths and the standard deviations of C-segments' lengths on chromosomes 1, 9, 16 and Y obtained in this study are 1.26 +/- 0.23; 1.13 +/- 0.20; 0.85 +/- 0.14; 1.13 +/- 0.21 mcm respectively. Identical data on both sexes, as well as the normal distribution of C-segments' lengths confirm the idea that the length of the C-segment is a neutral characteristic in normal population.

Adolescent↗