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Ear oximetry: a noninvasive method for detection of patent foramen ovale: a study comparing dye dilution method and oximetry with contrast transesophageal echocardiography.

BACKGROUND AND PURPOSE: Patent foramen ovale (PFO) may play an important role as a risk factor for ischemic stroke and some other neurological conditions. There is a need for low-cost and noninvasive methods for the detection of PFO. This study evaluates the accuracy of two simple bedside tests, the dye dilution method and ear oximetry, in the detection of PFO. METHODS: Dye dilution curves and ear oximetry recordings with a noninvasive ear densitometer were obtained from consecutive cryptogenic stroke patients referred for contrast transesophageal echocardiography (TEE). All test results were blindly assessed for the presence of PFO. Sensitivity and specificity were calculated with TEE used as a reference method. kappa statistics were used to measure interrater agreement. RESULTS: Dye dilution curves were obtained from 67 patients. Dye dilution correctly diagnosed 35 of the 46 patients who had PFO in TEE and all the 21 patients without PFO. Thus, the sensitivity (95% CI) of the dye dilution method was 76% (61% to 87%) and its specificity 100% (84% to 100%). Ear oximetry was done on 83 patients. Oximetry correctly diagnosed 45 of the 53 patients who had PFO in TEE and all of the 30 patients without PFO. Thus, the sensitivity of ear oximetry was 85% (72% to 93%) and its specificity 100% (88% to 100%). The interrater agreement was excellent (kappa value 0.94 for dye dilution and 0.90 for oximetry). CONCLUSIONS: Dye dilution and oximetry are both sensitive and specific methods for the detection of PFO. Oximetry has the following primary advantages over the currently available diagnostic methods: it is noninvasive, safe, and inexpensive and causes no discomfort for the patient. We suggest that oximetry could be used as a first-line screening method for PFO in patients with cryptogenic stroke. Ear oximetry also has potential use in epidemiological studies.

Adult↗

The unlabeled antibody method: comparison of peroxidase-antiperoxidase with avidin-biotin complex by a new method of quantification.

Upon plotting of areas against optical densities in immunocytochemically stained tissue sections, hyperbolic curves were obtained which could be reduced to two straight lines, one representing variations in stained structures, and the other variations in background. The slopes of the stained structure lines reflected staining intensity independently of total area of stained structure in a section. The ratio of slopes of the stained structure and background lines reflected immunocytochemical sensitivity. A comparison of the peroxidase-antiperoxidase (PAP) method with the avidin-biotin complex (ABC) method showed that at usual antibody dilutions the PAP method was much more sensitive than the ABC method, while at impractically high antibody dilutions it was moderately more sensitive. Once sufficient dilutions of antibodies were reached, staining intensities dropped sharply with the PAP method. On the other hand, the dilution curves were flat with the ABC method. The ABC method consequently appeared unsuitable for estimating variations in concentration of antigen or for distinguishing high or low concentrations of antigen. The ABC method provided a stain for myelin even in the absence of any antibodies.

Animals↗

Comparison of bedside measurement of cardiac output with the thermodilution method and the Fick method in mechanically ventilated patients.

INTRODUCTION: Bedside cardiac output determination is a common preoccupation in the critically ill. All available methods have drawbacks. We wished to re-examine the agreement between cardiac output determined using the thermodilution method (QTTHERM) and cardiac output determined using the metabolic (Fick) method (QTFICK) in patients with extremely severe states, all the more so in the context of changing practices in the management of patients. Indeed, the interchangeability of the methods is a clinically relevant question; for instance, in view of the debate about the risk-benefit balance of right heart catheterization. PATIENTS AND METHODS: Eighteen mechanically ventilated passive patients with a right heart catheter in place were studied (six women, 12 men; age, 39-84 years; simplified acute physiology scoreII, 39-111). QTTHERM was obtained using a standard procedure. QTFICK was measured from oxygen consumption, carbon dioxide production, and arterial and mixed venous oxygen contents. Forty-nine steady-state pairs of measurements were performed. The data were normalized for repeated measurements, and were tested for correlation and agreement. RESULTS: The QTFICK value was 5.2 +/- 2.0 l/min whereas that of QTTHERM was 5.8 +/- 1.9 l/min (R = 0.840, P < 0.0001; mean difference, -0.7 l/min; lower limit of agreement, -2.8 l/min; upper limit of agreement, 1.5 l/min). The agreement was excellent between the two techniques at QTTHERM values <5 l/min but became too loose for clinical interchangeability above this value. Tricuspid regurgitation did not influence the results. DISCUSSION AND CONCLUSIONS: No gold standard is established to measure cardiac output in critically ill patients. The thermodilution method has known limitations that can lead to inaccuracies. The metabolic method also has potential pitfalls in this context, particularly if there is increased oxygen consumption within the lungs. The concordance between the two methods for low cardiac output values suggests that they can both be relied upon for clinical decision making in this context. Conversely, a high cardiac output value is more difficult to rely on in absolute terms.

Adult↗

A near-infrared method for the assay of cineole in eucalyptus oil as an alternative to the official BP method.

Thirty different eucalyptus oil samples were scanned on the FOSS NIRSystems 6500 Rapid Content Sampler using a reflectance vessel as sample presentation method. The cineole content of each sample was determined by the BP method and these reference data were used to construct two calibration equations for cineole content in the oils using Vision software. The mean accuracy for the NIR method differed by 1.01% or less, and the mean bias by +/-0.33% or less, compared with the BP method. Calculation of the 95% confidence intervals for the slope and intercept of plots of NIR predicted values against BP method reference values showed that there was no evidence of fixed or relative systematic errors. Tests for short-term and intermediate repeatability were conducted. The standard deviation was 0.83% w/w or less and the coefficient of variation was 1.11% or less. The confidence intervals for both short-term and intermediate repeatability overlapped with that for the BP method, suggesting that there was no evidence for a difference in values obtained by the BP and NIR methods. The range of cineole contents used in the calibrations was extended by incorporating five samples of eucalyptus oil spiked with cineole, and five samples of two essential oils known to have a lower cineole content than eucalyptus oil, to give a range of 52.5 to 99.0% w/w. The mean accuracy decreased to an error of 1.26% or less and the bias to +/-0.50% or less. Again, confidence intervals suggested there was no evidence for fixed or systematic errors in the NIR calibrations. We propose that NIR spectroscopy could be used as an alternative method for the determination of cineole content in eucalyptus oils.

Cyclohexanols↗

A candidate reference method for the determination of uric acid in serum based on high performance liquid chromatography, compared with an isotope dilution-gas chromatography-mass spectrometer method.

A method based on isocratic high performance liquid chromatography (HPLC) with UV detection at 292 nm is proposed as a candidate reference method for the determination of uric acid. Data obtained by this method are compared with those from an isotope dilution-gas chromatography-mass spectrometric method (ID-GC-MS), using [1,3-15N2]uric acid as internal standard and selected mass detection at m/z = 456 and m/z = 458. The inaccuracy of the ID-GC-MS method is maximally 0.4% for NBS-SRM-909 control sera with a concentration of 483 mumol/l. The coefficient of variation between days is 0.26%-0.80% and 0.37-0.90% for 14 control sera from other suppliers. The maximum bias of the HPLC method is 0.6%, and the coefficient of variation between days is 0.31%-0.65% for NBS-SRM-909 control sera. The coefficient of variation between days for the other 14 control sera tested is 0.35%-0.66%. Comparison of the HPLC method with the reference ID-GC-MS method resulted in a coefficient of correlation of r = 0.9998 (n = 14). The concentration of uric acid in the tested control sera ranged from 160 to 624 mumol/l.

Chromatography, High Pressure Liquid↗

A method for the simultaneous determination of creatinine and uric acid in serum by high-performance-liquid-chromatography evaluated versus reference methods.

A high performance liquid chromatography (HPLC) with isocratic ion-pair-reversed-phase separation and simultaneous UV-detection at 232 nm and 292 nm is proposed as a method for the simultaneous determination of uric acid and creatinine in serum. The only sample preparation required is an appropriate dilution with the eluent and membrane filtration on non-adsorbent 0.2 micron membrane-filtration-devices. The inaccuracy of the method has been determined for NIST-SRM-909 (n = 10) and was + 0.5% for creatinine as well as for uric acid. The imprecision in this case was 0.8% for both analytes. The within-run imprecision for creatinine/uric acid was 0.4-0.5%/0.2-0.4% in the case of standards and 0.6-0.8%/0.4-0.7% in the case of serum-pools. The between-run imprecision for creatinine/uric acid obtained from serum pools was 0.8-1.1%/0.7-1.0%. The results for creatinine have been compared to those from an isotope dilution-gas chromatography-mass spectrometry using [13C, 15N2]creatinine as internal standard and selected mass detection at m/e = 329 and m/e = 332. The results for uric acid have been compared to an HPLC-method published previously (Kock R et al. J Clin Chem Clin Biochem 1989; 27:157-62). The method comparisons (n = 55) for the new combined method presented versus the reference method for creatinine and the candidate reference method for uric acid resulted in coefficients of correlation of r = 1.000 for both analytes. The new combined method presented is useful for the analysis of patient samples where the classical photometric procedures do not give reliable results, as often observed in monitoring after transplantation surgery.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromatography, High Pressure Liquid↗

The empirical bias of estimates by restricted maximum likelihood, Bayesian method, and method R under selection for additive, maternal, and dominance models.

Bayesian analysis via Gibbs sampling, restricted maximum likelihood (REML), and Method R were used to estimate variance components for several models of simulated data. Four simulated data sets that included direct genetic effects and different combinations of maternal, permanent environmental, and dominance effects were used. Parents were selected randomly, on phenotype across or within contemporary groups, or on BLUP of genetic value. Estimates by Bayesian analysis and REML were always empirically unbiased in large data sets. Estimates by Method R were biased only with phenotypic selection across contemporary groups; estimates of the additive variance were biased upward, and all the other estimates were biased downward. No empirical bias was observed for Method R under selection within contemporary groups or in data without contemporary group effects. The bias of Method R estimates in small data sets was evaluated using a simple direct additive model. Method R gave biased estimates in small data sets in all types of selection except BLUP. In populations where the selection is based on BLUP of genetic value or where phenotypic selection is practiced mostly within contemporary groups, estimates by Method R are likely to be unbiased. In this case, Method R is an alternative to single-trait REML and Bayesian analysis for analyses of large data sets when the other methods are too expensive to apply.

Animals↗

[Utility of the paraffin-embedded section method on the detection of estrogen receptor from breast cancer tissues--comparison of the paraffin-embedded section method (6F11 and 1D5) with frozen section (H222) and dextran-coated charcoal (DCC) ones].

From March 1988 to December 1990, we detected estrogen receptor (ER) from breast cancer tissues with the dextran-coated charcoal (DCC) method. From September 1990 to March 1998, we repeated the above with the frozen section method (cloneH222; DAINABOT). With the paraffin-embedded section method using two antibodies of anti-ER (clone6F11; NOVOCASTRA and clone 1D5; MBL), we examined the ER of the same 185 primary breast cancer tissues. We had already detected these tissues with the frozen section method, and we also applied the same procedure for the 43 primary breast cancer tissues which had already been detected with the DCC method. We compared these data. The positive rates of ER with DCC, H222, 6F11 and 1D5 were 49%, 53%, 53% and 52% respectively, which were within the reported range. The accuracy between H222 and 6F11 which was calculated as the percentage that were in positive and negative concordance by the two methods, was 88%. The accuracies between H222 and 1D5, between 6F11 and 1D5, between DCC and 6F11, between DCC and 1D5 were respectively 89%, 96%, 79%, and 77%. Although the accuracy between DCC and the paraffin-embedded section was not necessarily high, we obtained a higher concordance between the frozen and paraffin-embedded sections. The highest concordance existed between 6F11 and 1D5. The 30% of negative cases with DCC were positive with paraffin-embedded section. Among these methods of ER detection from breast cancer tissues, the paraffin-embedded section method seemed to be the most useful.

Breast Neoplasms↗

Transmembrane topology prediction methods: a re-assessment and improvement by a consensus method using a dataset of experimentally-characterized transmembrane topologies.

We selected 10 transmembrane (TM) prediction methods (KKD, TMpred, TopPred II, DAS, TMAP, MEMSAT 2, SOSUI, PRED-TMR2, TMHMM 2.0 and HMMTOP 2.0) and re-assessed its prediction performance using a reliable dataset with 122 entries of experimentally-characterized TM topologies. Then, we improved prediction performance by a consensus prediction method. Prediction performance during re-assessment and consensus prediction were based on four attributes: (i) the number of transmembrane segments (TMSs), (ii) the number of TMSs plus TMS-position, (iii) N-tail location and (iv) TM topology. We noted that hidden Markov model-based methods dominate over other methods by individual prediction performance for all four attributes. In addition, all top-performing methods generally were model-based. Among prokaryotic sequences, HMMTOP 2.0 solely topped among other methods with prediction accuracies ranging from 64% to 86% across all attributes. However, among eukaryotic sequences, prediction performance for all the attributes was relatively poor compared with prokaryotic ones. On the other hand, our results showed that our proposed consensus prediction method significantly improved prediction performance by, at least, an additional nine percentage points particularly among prokaryotic sequences for the number of TMS (84%), number of TMS and position (80%), and TM topology attributes (74%). Although our consensus prediction method improved also the prediction performance among eukaryotic sequences, the obtained accuracies for all attributes were relatively lower than that obtained by prokaryotic counterparts particularly for TM topology.

Cell Membrane↗

The intercept method: a novel method for establishing consistency of M-wave recruitment curves.

The stability of the M-wave is an important component of experimental H-reflex methodology. Despite this importance, there is inconsistency in H-reflex literature on the most valid method of M-wave stability analysis. Further, there is currently no specific method for establishing the stability of an M-wave recruitment curve across various trials within an experiment. Therefore, the aim of this study was to investigate the most appropriate method of M-wave stability analysis for use with the recruitment curve methodology. Twenty-five healthy subjects participated in the study. Four M-wave recruitment curve recordings were made in various static positions that imposed stretch on the posterior structures of the back and leg. Four methods of post-data collection M-wave stability analysis were compared. Although on visual inspection, there was clear evidence of marked alterations to the M-wave recruitment curves between trials in some subject's data, analysis of variance of the Ms/p and Mmax found no significant difference. Evaluation of the percent deviation in Mmax found nine subjects with greater than ten percent deviation in their maximum M-wave across the four trials. The intercept method that utilises analysis of the 95% confidence interval of the intercept of the M-wave recruitment curve slope, excluded eight subjects that demonstrated variation. Comparison of the percent deviation and the intercept method revealed that the intercept method was the most appropriate method for M-wave stability analysis in conjunction with the recruitment curve methodology.

Adult↗

Comparison of two methods for the estimation of urea kinetics and introduction of a third simplified method.

It has been claimed that computed urea kinetic (UK) modelling in hemodialysed patients, for the estimation of protein intake, leads to an overestimation of protein catabolic rate (PCR). In the present study, three different methods of kinetic modelling for the determination of PCR and Kt/V are compared in 24 patients. The first method was the direct quantification method (DDQ) based on the collection of all urea eliminated from the body. The first computed method (ICMI) was the urea kinetic modelling method as described by Sargent. Dialyzer clearances were measured directly and not estimated by theoretical extrapolation. The second computed method (ICMII) is based on the indirect calculation of urea distribution volume (Vu), according to Watson, and of dialyzer clearances from this Vu and from pre- and post-dialysis urea concentrations. All three methods resulted in PCR's that were not significantly different (DDQ: 1.03 +/- 0.19; ICMI: 1.04 +/- 0.22; ICMII: 1.08 +/- 0.25 mg/Kg BW.24 hrs; p greater than 0.05). When the results were correlated, the following results were obtained: ICMI vs ICMII: r = 0.89, p less than 0.001; ICMI vs DDQ: r = 0.68, p less than 0.01; DDQ vs ICMII: r = 0.78, p less than 0.001. Intermutual comparison of Kt/V values resulted in virtually identical results, especially when comparing ICMI and ICMII, where the regression line equalled the identity line. In conclusion, all methods seem equally reliable in determining mean PCR and Kt/V. Our data, obtained with directly measured dialyzer urea clearances, do not confirm the earlier held opinion that computed modelling results in an overestimation of PCR.(ABSTRACT TRUNCATED AT 250 WORDS)

Female↗

Light has a greater effect on direct bilirubin measured by the bilirubin oxidase method than by the diazo method.

We compared the effect of light on direct-reacting bilirubin (DBIL) measurement by the bilirubin oxidase (EC 1.3.3.5; BOX) method and by the Jendrassik-Gróf diazo method. DBIL concentrations determined by the BOX method in the sera of hyperbilirubinemic infants treated with phototherapy yielded falsely higher values than those by the direct diazo method. A similar tendency was noted when DBIL concentrations in infants' sera irradiated with light in vitro were determined by both methods, although by HPLC none of these sera had detectable DBIL (i.e., conjugated plus delta bilirubin). In general, DBIL concentrations after photoirradiation remained unchanged when measured by the diazo method, but significantly increased when the BOX method was used. Indeed, photoirradiation gave rise to material that acted like a photobilirubin product, which was oxidized at pH 3.7 and therefore was measured as DBIL. Such false increases in DBIL values generated by the BOX method may have clinical diagnostic implications in monitoring jaundiced neonates and in differentiating between physiological jaundice and incipient pathological jaundice.

Azo Compounds↗

A new commercial method for the enzymatic determination of creatinine in serum and urine evaluated: comparison with a kinetic Jaffé method and isotope dilution-mass spectrometry.

We evaluated a new, simple, enzymatic kinetic method from Wako Chemicals GmbH in comparison with a kinetic Jaffé method by using isotope dilution-mass spectrometry (ID-MS) as a reference method. An ID-MS-calibrated serum standard was used. Both the enzymatic and the Jaffé method correlated well with ID-MS, except for sera with high concentrations of bilirubin. Ethyl acetoacetate, acetone, and glucose in serum interfered somewhat with the Jaffé method but not with the enzymatic method. We conclude that the present enzymatic method has merit as compared with a Jaffé method for routine work, but is more expensive.

Amidohydrolases↗

[Clinical methods for evaluating infarct size and its anatomic correlations. Study carried out in 193 cases. III. Comparison of data on the evaluation of the infarct size using the QRS score and a method of maximal creatine kinase determination in the serum].

AIMS: To compare two methods concerning the clinical evaluation of infarct size--one using a QRS score, the other based on peak Ck values--applied to the same population. CONCEPT AND PLACE OF THE STUDY: to determine--based on previously established correlations between a QRS score and the anatomic total infarct size on one hand, and between the peak CK values and the anatomic recent infarct size on the other hand--which myocardial infarction subgroup constitutes the best indication for each method. The study took place in a Coronary Care Unit of a Central Hospital. MATERIAL AND METHODS: 193 patients who died successively of acute myocardial infarction through out 4 years were studied. After establishing the exclusion critéria, the QRS score was calculated according to the method of Selvester modified by Wagner, and peak CK values were evaluated. Infarct size, either recent or old, was determined by means of an anatomical method developed by the authors and based on Hackel's and Alonso's previous works. Correlations were established between data from each clinical method and those from the anatomical method. Several myocardial infarction subgroups were considered for comparison of the correlations found in each subset. RESULTS AND CONCLUSIONS: As long as QRS score was regarded, significant correlations were found between the evaluation by QRS score and anatomical infarct size in the subgroups of patients with severe pump failure, prior myocardial infarction, or total loss of ventricular muscle mass of at least 20 percent; however, a significant correlation was missing when the whole myocardial infarction group was taken into consideration. When peak CK value were considered, a weak significant correlation was found between the evaluation by enzyme determination and anatomical infarct size in the whole group of patients, but a stronger correlation was present in the subgroups of patients with survival longer than 24 hours, anterior wall myocardial infarction, free wall cardiac rupture, or first acute myocardial infarction. In conclusion the choice of the method to be used in the clinical evaluation of infarct size should take into account the type of population beeing studied, and follow the results obtained in different myocardial infarction subgroups as mentioned above.

Creatine Kinase↗

[New method for preventing spinal headache (prophylactic blood patch method)].

After spinal anesthesia, early ambulation frequently caused spinal headache. If the patient was discharged on the day of the operation, spinal headache occurred very frequently. With epidural anesthesia, if the dura was erroneously punctured with the needle, headache often occurred. In order to prevent spinal headache, we tried the Blood Patch Method during operation (Prophylactic Blood Patch Method). Epidural catheterization and spinal tap were simultaneously performed. The epidural catheter was threaded 3 cm cephalad in the epidural space. Five ml Venous blood of the patient was injected through the epidural catheter (Blood Patch Method). Epidural puncture was performed one spinal segment cephalad (A Method) or one spinal segment cauded (B Method) from the site of spinal puncture. The prophylactic Blood Patch Method was very effective in preventing spinal headache after the dural puncture. Especially B Method, in which blood was injected near the dural pore, was much more effective than A Method.

Anesthesia, Obstetrical↗

[Evaluation of assay methods for isepamicin sulfate (HAPA-B) in body fluids. Bioassay, HPLC and EIA methods].

Assay methods including microbiological assay (bioassay), high performance liquid chromatography (HPLC) and enzyme immunoassay (EIA) for isepamicin sulfate (HAPA-B), a new aminoglycoside antibiotic, in body fluids were studied. The most suitable bioassay method was double layer agar-well method using Bacillus subtilis ATCC 6633 as the test organism on plate consisting of a seed-layer of nutrient agar at pH 8.0 and a base-layer of nutrient agar supplemented with 0.4% sodium chloride. Sensitivities in bioassay, HPLC and EIA methods for plasma concentration were 0.08 microgram/ml, 0.20 microgram/ml and 0.05 microgram/ml, respectively. Plasma and urinary concentrations after intramuscular administration of HAPA-B at the dose of 200 mg to healthy volunteers were measured with these 3 methods. The HPLC and the EIA methods yielded values which compared favorably to the bioassay method. Using the bioassay method, HAPA-B levels in human plasma and urine samples were found to be stable at least for 15 days at -20 degrees C.

Aminoglycosides↗

Ten electrophoretic methods compared with a selected method for quantifying lactate dehydrogenase isoenzymes in serum.

Using the Selected Method of McKenzie and Henderson (Selected Methods Clin Chem 1983;10:59-67) as a reference method, we compared the performance of 10 commercially available methods for determination of lactate dehydrogenase (LD, EC 1.1.1.27) isoenzymes. Results were expressed as percentage of total LD activity, as determined with two different types of densitometers shown to have an average difference less than 1.4% for each isoenzyme. All methods gave generally comparable results, as judged by Bland-Altman plots and correlation analyses. However, in general, estimates by the commercial methods for LD-1, LD-2, and LD-3 were lower, and for LD-4 and LD-5 were higher than with the Selected Method. The overall CV was less than 20% for all methods and isoenzymes, except for LD-4 and LD-5 by the Beckman Paragon, Helena LD-VIS, Gel LDH, Gel PC, and Iso Dot, Gelman LDH Isozyme, and Sebia Hydragel assays, for which it was greater than 20%. Overall, accuracy was best with the Helena Iso Dot and LD-VIS assays, followed by the Corning LD Flur assay; accuracy was poorest with the Gelman LDH Isozyme, Sebia Hydragel, and Beckman Paragon assays.

Electrophoresis↗

[The evaluation of anti-native dna antibodies. Comparison of two methods of dosage, study of reproducibility, and diagnostic value of each method].

The duplicatibility of dosing anti-native (double-stranded) DNA antibodies using Farr's radioimmunological method (RIE) and indirect immunofluorescence on Crithidia luciliae (IF-CL) has been demonstrated in the literature and from a multicenter study undertaken in 10 immunological laboratories. The duplicatibility of tests run at the same time and also between tests done at different intervals by RIE in our laboratory was better than that by IF-CL. The coefficients of maximum change were respectively 3% (variation in results during one run of the test), and 6% (variation in results between different runs of the test) for the RIE method and 34% (variation during one run) and 38% (variation between different runs) using the IF-CL method. This multicenter study showed that the duplicatibility of Farr's radioimmunological test was good among the different laboratories, only antibody levels near the upper limit of normal giving some discordant results were found in 91%, the discordant results being due to falsely-positive or falsely-negative tests, and sometimes also to the differing affinity of the anti-native DNA antibodies in the two methods. The sensitivity of both methods in making a diagnosis of systemic lupus erythematosus from the literature varies from 60 to 98% for the RIE method and from 44 to 98% for the IF-CL method. The predictive value of a positive test between these two methods varies from 65 to 100% depending on the series, which means that there are other conditions than systemic lupus erythematosus (SLE) (rheumatoid arthritis, Sjögren's syndrome, hepatitis...) where anti-native DNA antibodies can be found.

Adult↗