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Calculation of standard transformed Gibbs energies and standard transformed enthalpies of biochemical reactants.

The standard Gibbs energies of formation and standard enthalpies of formation of species involved in biochemical reactions are used to calculate standard transformed Gibbs energies of formation and standard transformed enthalpies of formation of 53 reactants (sums of species) at 298.15 K, pH 7, and ionic strengths of 0, 0.1, and 0. 25 M. The standard transformed Gibbs energies of formation are used to calculate apparent equilibrium constants K' for 22 biochemical reactions for which apparent equilibrium constants have been determined close to these conditions. This comparison is generally satisfactory given the differences in experimental conditions. The transformed formation properties for the 53 reactants make it possible to calculate transformed formation properties for other reactants involved in biochemical reactions with some of these reactants. This is illustrated by calculating standard transformed Gibbs energies of formation for 11 more reactants without information on the standard Gibbs energies of formation of the species. The list of 64 reactants for which standard transformed Gibbs energies of formation are presented can be considerably extended. The use of tables of standard transformed Gibbs energies of formation to store information on apparent equilibrium constants is more efficient than simply storing apparent equilibrium constants because a reactant can be looked up in a table and may be involved in hundreds of reactions. The effects of magnesium ions on several reactions involving ATP are calculated. The advantages of using enzyme-catalyzed reactions for determining thermodynamic properties of complicated molecules in aqueous solution are discussed.

Biochemistry

Setting standards and defining quality of performance in the validation of a standardized-patient examination format.

PURPOSE: To evaluate whether written standards increase the reproducibility of a physician-facilitated station in an objective structured clinical examination (OSCE) designed to assess history, physical-examination, and communication skills. METHOD: The OSCE examination at the University of Texas Medical Branch-Galveston consists of ten eight-minute stations. Six of these stations consist of three History, Physical-examination, Problem-solving, and Plan (HPPP) station pairs. Each existing clinical-problem HPPP station was given to two content experts to develop standards for faculty rating scales appropriate for the evaluation of third-year medical students. Three pairs of faculty members were used to determine interrater reliability by scoring videotapes of three HPPP stations' presentation and problem-solving components. Faculty pairs scored tapes of 15 students without using standards and tapes of 15 students using the standards developed. Differences between the reliabilities without and with the standards were tested for significance using Fisher's R to Z transformation. The reproducibility and standard error of measurement (SEM) were extrapolated for increasing amounts of testing time. The HPPP component scores were also correlated with the written examination scores and preceptors' ratings. Data were obtained from the three HPPP stations used in the 1995-96 internal medicine clerkship SP examination. RESULTS: In all, 196 students completed the OSCE examination. The standards developed improved interrater reliability and reached statistical significance (p < .01) for one HPPP station. Reproducibility for the presentation and problem-solving components of the HPPP stations were > .80 after five hours of testing. The problem-solving component correlated at .37 and .19 with written examinations and with ward grades, respectively. CONCLUSION: The data from this study suggest that standards increase the reproducibility of presentation and problem-solving components of an OSCE to a level as high as, or higher than, that associated with the history, physical-examination, and communication components of traditional standardized-patient examinations.

Clinical Clerkship

Standardization of Factor VIII. I. Calibration of British Standards for Factor VIII clotting activity.

Calibration of successive British Standards for Factor VIII clotting activity against the International Standard (concentrate) has brought to light substantial discrepancies among laboratories and between assay methods. These discrepancies were less in assays of concentrate standards than in calibration of plasma standards. Standardization of reagents in the two-stage assays substantially improved agreement among laboratories using this method. Standardization of the phospholipid reagent and haemophilic substrate separately had little effect on variation among laboratories performing one-stage assays. Standardization of reagents did not alter the basic discrepancy (approximately 20%) between the one-stage and two-stage assay methods. Omission of the aluminium hydroxide adsorption step from the two-stage method reduced the discrepancy between the two methods to less than 10%. Discrepancies did not occur when one plasma standard was assayed against another. Improvements in the stability of VIII:C in freeze-dried plasma now make it feasible to establish a long-term plasma reference standard.

Adsorption

Standardization of Factor VIII-IV. Establishment of the 3rd International Standard for Factor VIII: C concentrate.

An international collaborative study was carried out to establish a replacement for the current (2nd) international standard for Factor VIII:C, concentrate. Twenty-six laboratories took part, of which 17 performed one-stage assays, three performed two-stage assays and six used both methods. The proposed new standard, an intermediate purity concentrate, was assayed against the current standard, against a high-purity concentrate and against an International Reference Plasma, coded 80/511, previously calibrated against fresh normal plasma. Assays of the proposed new standard against the current standard gave a mean potency of 3.89 iu/ampoule, with good agreement between laboratories and between one-stage and two-stage assays. There was also no difference between assay methods in the comparison of high-purity and intermediate purity concentrates. In the comparison of the proposed standard with the plasma reference preparation, the overall mean potency was 4.03 iu/ampoule, but there were substantial differences between laboratories, and the two-stage method gave significantly higher results than the one stage method. Of the technical variables in the one-stage method, only the activation time with one reagent appeared to have any influence on the results of this comparison of concentrate against plasma. Accelerated degradation studies showed that the proposed standard is very stable. With the agreement of the participants, the material, in ampoules coded 80/556, has been established by the World Health Organization as the 3rd International Standard for Factor VIII:C, Concentrate, with an assigned potency of 3.9 iu/ampoule.

Antigens

Analysis of medical screening and surveillance in 21 Occupational Safety and Health Administration standards: support for a generic medical surveillance standard.

Twenty-one Occupational Safety and Health Act (OSHA) standards were identified which contain medical service provisions intended to help in the identification and control of harmful health effects of workplace exposures. The utility and effectiveness of these provisions have not previously been evaluated. All 21 standards were reviewed and assigned numerical scores for each of 24 potential medical program elements. Several of these elements were combined to calculate Quality Control, Screening Utility, and Surveillance Utility scores for each standard. Total scores varied greatly, suggesting a lack of consistency and uniformity which was even more obvious when the actual regulatory language was examined. The mean Quality score was only 26% of potential points. Seventeen of 21 standards received less than half the total possible Quality score. When arrayed on a two by two matrix only two standards scored above 50% for both Screening and Surveillance Utility. It was concluded that the medical service provisions in OSHA standards are lacking in consistency and coherence. Two major shortcomings are the lack of quality control elements and the absence of surveillance features which would permit medical program results to be utilized for prevention activities including the identification and control of workplace hazards. A generic occupational medical surveillance standard could address these current weaknesses. Elements of such a generic standard are proposed.

Data Collection

A new twist in US health care data standards development: adoption of electronic health care transactions standards for administrative simplification.

To reduce the costs of common administrative transactions, health plans, payers and providers encouraged the US Congress to legislate administrative simplification of specific electronic health data transactions. The Health Insurance Portability and Accountability Act (HIPAA) of 1996 created incentives for a public-private partnership to develop and implement standards for the uniformity of health care data used in electronic administrative health transactions and standards for the privacy and security of individually identifiable health information. The standards' requirements of HIPAA and how they have been met by the US government hold promise for accelerating the uniform adoption of standards developed by accredited standards developing organizations. The transactions designated by Congress, the process of choosing the standards in the Department of Health and Human Services, the principles that guided these choices, and the actual choices, are presented here. A successful partnership for administrative health data standards can pave the way for success in clinical health data standards and their application in computer-based patient record systems.

Computer Security

The effect of 'standard drink' labelling on the ability of drinkers to pour a 'standard drink'.

Australia's National Health Policy on Alcohol has recommended that beverage containers be labelled so that alcohol content is 'readily understandable by the public'. Health promotion to increase the responsible use of alcohol now relies extensively on the concept of a standard drink--usually defined as 10 g of ethyl alcohol. Numerous difficulties confront a drinker who wishes to apply the standard drink system to monitor alcohol intake. This report describes a series of experimental tests of the proposal that these difficulties are minimised if alcohol containers have their alcohol content indicated in terms of standard drinks in addition to the usual percentage alcohol by volume method. Subjects were drinkers recruited from a Perth shopping mall and were tested only on beverage types they had consumed within the previous week. They were required to pour what they judged to be a single standard drink from a 750 ml bottle of either wine or beer. Beer drinkers achieved greater accuracy in this task when the bottles had standard drink labels, even when the glass size and beverage strength were varied. Wine drinkers had equal difficulty with this task whether standard drink or percentage labels were used. The addition of a 'ladder' up the side of a wine bottle with graduations in standard drinks would be necessary for wine drinkers to achieve a high level of accuracy. We conclude that labelling drink containers with their alcohol content in terms of standard drinks would better equip all drinkers to follow the advice of health educators.

Adolescent

Standardization of protein C in plasma: establishment of an international standard.

An international collaborative study, involving 18 laboratories, was carried out to establish an international standard for protein C in plasma. The proposed standard, which consisted of a freeze-dried ampouled plasma preparation coded 86/622, was assayed against fresh normal plasma and the participants' local standards. Protein C activity assays were placed in four groups, depending on the method of activation and detection of protein C. The combined potencies (units per ampoule) for the proposed international standard were: thrombin activation/clotting assays, 0.86; thrombin activation/chromogenic assays, 0.81; snake venom activation/clotting assays, 0.81 and snake venom activation/chromogenic assays, 0.82. Measurement of protein C antigen gave potency estimates of 0.81 and 0.82 unites per ampoule for the Laurell electroimmunoassay and ELISA techniques, respectively. The good agreement in potency estimates between the different methods indicates that the overall combined figure (226 assays) for the international standard of 0.82 international units per ampoule should serve for all methods. Accelerated degradation studies have indicated that the standard should be suitably stable when stored at -20 degrees C. The freeze-dried plasma 86/622 has been established by the WHO Expert Committee on Biological Standardization as the 1st International Standard for Protein C in Plasma; with an assigned unitage of 0.82 international units per ampoule.

Blood Coagulation Tests

Reporting standards for endovascular surgery: should existing standards be modified for newer procedures?

Appropriate standards for reports on endovascular procedures are discussed in terms of whether existing standards suffice or should be revised. SVS/ISCVS and SCVIR reporting standards form the basis for recommendations. The recently revised SVS/ISCVS "recommended standards for reports dealing with lower extremity ischemia" serves appropriately for most outcome comparisons for occlusive disease, that is, where open surgical and endovascular procedures should be assessed by the same standards. Existing endovascular reporting standards are used to augment these where specific considerations pertinent only to endovascular procedures, such as description of lesion, are needed. In some aspects, such as the reporting of complications that are specific to endovascular procedures (eg, contrast-related, medication-related, device-related, and some entry site and deployment-related complications), recent SCVIR recommendations are endorsed. Although some of these apply, additional reporting standards are required for reporting the results of endograft treatment of aneurysmal disease. The former are identified, and the latter are taken from recent recommendations for reports on the endograft repair of infrarenal AAAs and experiences from recent endograft trials. Public disclosures, at professional meetings or press releases, of data lacking the objectivity and uniformity of these reporting standards, because of the time lag before firm data can reach peer-reviewed journals, may mislead and impede progress.

Forecasting

Standardization of high-resolution flow cytometric DNA analysis by the simultaneous use of chicken and trout red blood cells as internal reference standards.

Determination of nuclear DNA content by flow cytometry requires comparison with a reference standard. The use of external standards such as lymphocytes or granulocytes is time-consuming and inaccurate. Chicken red blood cells (CRBC) have a DNA content of 35% of the human diploid value and have been widely used as internal standard. The ratio calculated on the basis of the peak channel numbers of the standard and the sample and used to indicate the DNA content (DNA ratio) is, however, very sensitive to changes in the zero level adjustment of the flow cytometer. If two internal standards are used the DNA ratio becomes independent of the zero level. Rainbow trout red blood cells (TRBC) have a DNA content of 80% of human diploid cells. A mixture of CRBC and TRBC was prepared and stored in small aliquots at -80 degrees C. This mixture was added to the sample before staining. The day-to-day variation of the DNA ratio obtained by use of the two standards was smaller than that obtained by CRBC alone. The possibility of sex related differences in DNA content of CRBC and TRBC was examined. The results indicated that a new batch of standards should be tested against the old batch to avoid the introduction of a systematic error.

Animals

Rabies vaccine standardization: International Collaborative Study for the Characterization of the fifth International Standard for Rabies Vaccine.

A collaborative study was carried out to establish a replacement for the International Standard for Rabies Vaccine, the stocks of which are exhausted. Three rabies vaccines for human use derived from different rabies virus strains and prepared on different cell culture substrates were compared with the International Standard for Rabies Vaccine using in vivo and in vitro assay methods in a collaborative study involving 14 participants. The proposed fifth International Standard (PISRAV) which was derived from the same virus strain as the present international standard preparation, the Pitman Moore (PM) strain, was found to be approximately twice as potent relative to the International Standard in immunogenicity assays as in antigenicity assays. On the other hand another vaccine, derived from the LEP strain, was considerably more potent in antigenicity assays than in immunogenicity assays. The glycoprotein of the proposed replacement standard measured in antigenicity assays appeared to be stable at +37 degrees C for 245 days, whereas the immunogenicity of the proposed replacement vaccine was sensitive to this heat treatment and the vaccine lost 66% of its immunogenic potency. The results of this study indicate that the NIH protection test should continue to form the primary basis for potency assay of rabies vaccine as glycoprotein content does not appear to correlate with immunogenic potency for different types of vaccine. The vaccine coded PISRAV has been established as the fifth International Standard for Rabies Vaccine and a potency of 16 International Units of Rabies Vaccine (based on the immunogenicity assays) assigned to the contents of each ampoule. Each ampoule has also been assigned a unitage of 10 IU of PM Rabies Virus Glycoprotein and 135 IU of PM Rabies Virus Ribonucleoprotein.

Animals

Gene differences between the sex ratio and standard gene arrangements of the X chromosome and linkage disequilibrium between loci in the standard gene Drosophila pseudoobscura.

The Standard and Sex Ratio gene arrangements of the X chromosome of D. pseudobscura differ from each other in allele frequencies at the four X chromosome loci, esterase-5, adult acid phosphatase-6, phosphoglucomutase-1 and octanol dehydrogenase-3. The Standard arrangement which is the common arrangement in all populations is polymorphic at these loci in varying degrees, the geographically less widespread Sex Ratio arrangement has little polymorphism and is genically predominantly E-5(1.04) AP-6(-) Pgm1(1.0) ODH-3(1.0). The Sex Ratio arrangement from different populations is alike at all of the four loci, the Standard arrangement shows some gene frequency differences among populations. The Standard and Sex Ratio arrangements differ from each other by three inversions which suggests that the two arrangements are "old". Gene differences between these two chromosome arrangements can be explained due to differential natural selection of alleles in the Standard and Sex Ratio arrangments.-The order and percent recombination among these four loci in the Standard arrangement are: E-5-.294-AP-6-.335-Pgm-1-.024-ODH-3. The Standard X chromosomes from four different wild populations were analyzed for evidence of linkage disequilibrium between pairs of loci at these four loci. No evidence of linkage disequilibrium between pairs of loci was obtained. However, when linkages involving simultaneously three loci, E-5, AP-6 and Pgm-1 are considered, then significant departure from linkage equilibrium is observed.

Acid Phosphatase

[Contact eczema and "standard test"--do "International Standard Test Series" coincide with Austrian local characteristics?].

Testing with a "standard" group of substances has proved of great value when allergic contact dermatitis is suspected and the offending agent cannot be pinpointed by careful history-taking. But internationally standardized series do not take into account locally important and individual factors, changing consumer habits or economic aspects. We have, thus, compared the suitability of two standard series, the ICDRG-European and the DKG-German standard, for Austrian purposes. 1,478 patients with a clinical diagnosis of contact dermatitis were patch tested between 11/88 and 4/90 with the ICDRG allergens; the second collective of 505 patients was tested between 5/90 and 1/91 with the DKG series. Only 12 out of the 27 substances of the ICDRG series exceeded the critical incidence of 1%; 6 were below this limit but positive in more than 0.5% and 9 hardly ever aroused positive reactions. In the DKG series, 17 out of 23 substances surpassed the limit, 2 were positive in more than 0.5%, and 4 caused virtually no reactions. However, 4 allergens with frequent sensitization rates were included neither in the ICDRG nor in the DKG series and 2 others were missing in the over-dimensioned ICDRG standard. Our results demonstrate that internationally established standard series have to adapted for the local situation in Austria, and that a constant review is essential with replacement of allergens as necessary according to changing production techniques and consumer habits. Suggestions for an "Austrian standard patch test series" are proposed on the basis of our results but in spite of being helpful in general terms interpretation of such a test in the individual patient with eczema remains problematic.

Allergens

Replacement of the international standard for tetanus antitoxin and the use of the standard in the flocculation test.

Since 1935 the International Unit for Tetanus Antitoxin has been defined as the activity contained in a certain weight of the first International Standard for Tetanus Antitoxin. As stocks of this standard had become depleted, 11 laboratories in 8 countries were requested to participate in a collaborative assay of a preparation proposed as a replacement.The assay results were analysed and presented to the WHO Expert Committee on Biological Standardization in 1969 which established the preparation studied as the second International Standard for Tetanus Antitoxin and defined the International Unit for Tetanus Antitoxin as the activity contained in 0.03384 mg of the second International Standard for Tetanus Antitoxin. This definition would ensure the continuity of the size of this international unit.The analysis of the collaborative studies also showed that the second International Standard for Tetanus Antitoxin has suitable properties for use in the flocculation test for the determination of the antigen content of tetanus toxoids in Lf values. The designation Lf-equivalent is described and the problems relating to the use of this term for the expression of results of in vitro assays are analysed in relation to the use of international units for expressing results of in vivo assays. As the second International Standard for Tetanus Antitoxin has an in vivo/in vitro ratio of 1.4, the Lf-equivalent of this antitoxin is 1.4 times less than its unitage.

Animals