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PubMed1975

[Comparison of conventional methods for the determination of albumin, transferrin and ceruloplasmin in serum with immunological reference methods (author's transl)].

Radial immunodiffusion, which is a presently accepted reference method, was compared with the mechanized immunoprecipitation. Each of these specific methods was compared with two normal clinical-chemical methods for the determination of albumin and transferrin, and an enzymic method for the determination of ceruloplasmin. The correlation coefficients, the mean slope for regression, and the percentage variation of the average values from those of the average values of the reference methods are presented. In the determination of albumin and ceruloplasmin, radial immunodiffusion and mechanized immunoprecipitation give the same results. Surprisingly, these two specific methods give different results in the determination of transferrin. In as far as there is no doubt concerning the accuracy of the mechanized immunoprecipitation, this method is recommended, since it also has the advantage of being less demanding in cost and time.

Autoanalysis↗
PubMed1983

Measurement of functional residual capacity during mechanical ventilation. Comparison of a computerized open nitrogen washout method with a closed helium dilution method.

The functional residual capacity of patients treated with mechanical ventilation was measured with two methods: the closed helium dilution method and the open nitrogen washout method with a computerized system. Measurements of FRC were made with and without PEEP. The results obtained did not show significant differences between the methods. Additionally we checked the accuracy of the open nitrogen washout method to measure FRC of patients and healthy subjects breathing high concentrations of oxygen which was the same as breathing air. Both methods were equally reliable and safe when measuring the FRC of patients on mechanical ventilation but the nitrogen method was easier and quicker.

Computers↗
PubMed1996

Rational experimental design for bioanalytical methods validation. Illustration using an assay method for total captopril in plasma.

Generally, bioanalytical chromographic methods are validated according to a predefined programme and distinguish a pre-validation phase, a main validation phase and a follow-up validation phase. In this paper, a rational, total performance evaluation programme for chromatographic methods is presented. The design was developed in particular for the pre-validation and main validation phases. The entire experimental design can be performed within six analytical runs. The first run (pre-validation phase) is used to assess the validity of the expected concentration-response relationship (lack of fit, goodness of fit), to assess specificity of the method and to assess the stability of processed samples in the autosampler for 30 h (benchtop stability). The latter experiment is performed to justify overnight analyses. Following approval of the method after the pre-validation phase, the next five runs (main validation phase) are performed to evaluate method precision and accuracy, recovery, freezing and thawing stability and over-curve control/dilution. The design is nested, i.e., many experimental results are used for the evaluation of several performance characteristics. Analysis of variance (ANOVA) is used for the evaluation of lack of fit and goodness of fit, precision and accuracy, freezing and thawing stability and over-curve control/dilution. Regression analysis is used to evaluate benchtop stability. For over-curve control/dilution, additional to ANOVA, also a paired comparison is applied. As a consequence, the recommended design combines the performance of as few independent validation experiments as possible with modern statistical methods, resulting in optimum use of information. A demonstration of the entire validation programme is given for an HPLC method for the determination of total captopril in human plasma.

Calibration↗
PubMed1990

Methods for estimating release rates during high frequency quantal secretion and for testing such methods.

The rate of spontaneous quantal release must be estimated in some investigations of synaptic transmission, even when frequencies are so high that individual quanta cannot be distinguished. An obvious method is to measure the time integrals of the summed MEPPs and then dividing this value by the integral of an average MEPP. The method was tested by recording miniature end-plate potentials (MEPPs) at frog neuromuscular junctions, counting the number of MEPPS, and then adding together records from the same junction to simulate high frequencies. The estimates from the integral method agreed well with the actual counts. The method can readily be used with a microcomputer and does not require stationary data. Methods based on fluctuation analysis were also used to estimate quantal frequencies, but they did not always give good estimates. This was not a thorough test of the fluctuation method, but an example of testing with MEPP data. The integral method might be reasonably reliable, but there are further potential complications, like changes in MEPP size and short-circuiting of the end-plate membrane, which may make it difficult to obtain reliable measurements of high frequency quantal secretion without voltage clamping and protocols that permit measurements of individual MEPCs during the course of the frequency measurements.

Animals↗
PubMed1996

Water turnover evaluated by the tritiated water method and by the nutritional method in European hedgehog (Erinaceus europaeus L.).

The total body water (TBW), the fractional water turnover rate (WTR), and the water flux rates (WFR) were studied for the first time by the tritiated-water method in 10 hedgehogs under laboratory conditions during the year. To validate this method, we compared the result to the nutritional method. The TBW varied inversely with body mass (BM) during both the active and the hibernation periods. The WTR varied from 16% to 21% during the hibernation period and after the reproduction. The higher values (from 22% to 26%) were recorded during the reproduction period and during the preparation for hibernation. The same variations in WFR were recorded during the active and hibernation period. The net gain or loss of water followed the gain and loss of BM. The tritiated water (THO) method has been validated by the fact that the same variations in WFR during each month have been measured by the THO method and the nutritional method. Moreover, the THO method can be used to estimate the changes in energy metabolism of hedgehogs in relation with the seasons.

Analysis of Variance↗
PubMed1996

Comparative study of bone mineral density estimated by various methods of single- and dual-energy quantitative computed tomography: the capability of the four-equation four-unknown method.

A dual-energy (DE) quantitative computed tomography (QCT) method, the four-equation four-unknown method (DEQCT 4E-4U), was assessed and compared to single-energy (SE) QCT and standard DEQCT (two-line method). The results of this study indicate that bone mineral density (BMD) was more accurately estimated by the present method than by the SEQCT or standard DEQCT techniques on the basis of a phantom study when a large fat content was present. The results of both the phantom study and a human study also showed that the present method corrected for fat in estimating BMD in the presence of high-fat content. These findings suggest that use of this method for estimating BMD can provide useful information in studies assessing the metabolic state of bone. We propose that CT numbers estimated from excised vertebral bone marrow can serve as a soft-tissue correction for the present method.

Adult↗
PubMed2026

An enhanced multisegment RT-PCR method for influenza A virus sequencing: Improved performance and reduced preparation time over traditional methods.

Influenza A viruses (IAVs) remain a major global health threat, affecting both human and animal populations. Whole-genome sequencing is essential for monitoring viral evolution, zoonotic transmission, and emerging variants. However, conventional RT-PCR methods often result in incomplete gene coverage, amplification biases, and reduced sequencing accuracy, particularly in clinical samples. We developed a robust In-house method for IAV full-genome sequencing using the Oxford Nanopore Technologies (ONT) long-read sequencing platform. This method integrates an in-house multisegment Reverse Transcription PCR (RT-PCR) method with a streamlined 2-pool primer design targeting all eight IAV gene segments. RNA extracted from clinical and stock virus samples was reverse-transcribed and amplified using Superscript IV-based chemistry, followed by magnetic bead purification to ensure high-quality amplicons. Sequencing libraries were prepared with the Native Barcoding Kit 24 (SQK-NBD114.24) and sequenced on R10.4.1 flow cells on the MinION MK1C device. Data analysis using the Iterative Refinement Meta-Assembler (IRMA) confirmed improved read depth, uniform coverage, and complete genome recovery. Compared to conventional methods, our In-House Multisegment 2-Pool (IH-MS2P) RT-PCR method generated higher numbers of matched read counts, minimized chimeric artifacts, and delivered superior genome coverage across human, swine, and avian isolates. This optimized RT-PCR method provides a high-performance, time-efficient, and portable solution for influenza genomics, demonstrating robust applicability even with clinical samples of low RNA yield.

Influenza A virus↗
PubMed1985

Light microscopical localization of enzymes by means of cerium-based methods. I. Detection of acid phosphatase by a new cerium-lead-technique (Ce-Pb-method).

Cerium-III-ions are more and more used as capturing reagent and opaque marker for the electron microscopic localization of a number of H2O2-generating enzymes as well as phosphohydrolases. Contrary to its advantages over common lead methods in the histochemical detection of enzyme activities at the electron microscopic level, cerium-based methods proved to be a failure for light microscopic investigations. Therefore, our cerium-based method for the ultrahistochemical detection of acid phosphatase was developed for further observations at the light microscopic level. The principle of that new Ce-Pb-method is the conversion of light microscopic not visible cerium phosphate into lead phosphate by the secondary capture reagent alkaline lead citrate. Finally, the lead phosphate can be visualized as lead sulfide in the section. The Ce-Pb-method in its finally proposed manner was compared with a common lead method and showed a range of advantages. Because of that fact, the new Ce-Pb-method is recommended for a broader use in histochemistry, e.g. for the light microscopic enzyme investigation parallel to ultrahistochemical preparations.

Acid Phosphatase↗
PubMed1986

The "Ring" method: a semi-empirical method to calculate dose distributions of irregularly shaped photon beams.

The "Ring" method provides a fast dose calculation and isodose presentation for photon beams with blocks. The method takes into account the change in scatter due to the blocks at each calculation point. Firstly, the dose in a point is calculated assuming that no blocks are present. Secondly, the scatter reduction caused by the blocks is calculated and subtracted. To determine the scatter reduction the irradiated surface is divided in concentric rings around a point at the surface at the intersection with a ray line between focus and calculation point. The scatter reduction caused by blocks for each ring is calculated. The effect of scatter for rings with an outer radius greater than 15 cm where the scatter contribution is less than 1.0% is neglected. Results of the method for 4 MV photons using eight rings are presented. Comparison of dose measurements with calculations in an arrow-shaped photon field showed maximum deviations of 4.0%, using the IRREG program of Cunningham, 6.5% using the BLKINP program of Schlegel, which is based on Clarkson's method, 5.0% using the method of Wrede and 2.2% using the "Ring" method. Contrary to the first two calculation programs, the programs using the last two calculation methods provide isodose lines dose values at points.

Radiation Dosage↗
PubMed2002

Improved method for quantifying the avicide 3-chloro-p-toluidine hydrochloride in bird tissues using a deuterated surrogate/GC/MS method.

A method using a deuterated surrogate of the avicide 3-chloro-p-toluidine hydrochloride (CPTH) was developed to quantify the CPTH residues in the gastrointestinal (GI) tract and breast muscle tissues in birds collected in CPTH-baited sunflower and rice fields. This method increased the range of a previous surrogate/gas chromatography/mass spectroscopy method from 0-2 to 0-20 microg/g in tissue samples and greatly simplified the extraction procedure. The modified method also sought to increase recoveries over a range of matrix effects introduced by analyzing tissues from birds collected in the field, where the GI tract contents would be affected by varying diet. The new method was used to determine the CPTH concentration in GI tract samples fortified with CPTH-treated rice bait to simulate the consumption of varying amounts of treated bait by two nontargeted bird species, pigeon (Columbia livia) and house sparrow (Passer domesticus). The new method was then used to examine the CPTH concentrations in the gizzard contents of the targeted bird species, red-winged black bird (Agelaius phoeniceus) and brown-headed cowbird (Molothrus ater), that were collected after feeding at a treated bait site. The method proved sufficiently sensitive to quantify CPTH in the breast muscle tissues and the gizzard contents of red-winged blackbirds and brown-headed cowbirds during an operational baiting program. The levels of CPTH determined for these birds in both tissue samples were determined to be highly correlated. The appearance of CPTH in the breast muscle tissue immediately after feeding was not anticipated. The potential secondary hazard posed by the targeted birds to potential scavengers and predators was also evaluated.

Animals↗
PubMed1998

Comparison of the proximal flow convergence method and the jet area method for the assessment of the severity of tricuspid regurgitation.

AIMS: To compare the value of the proximal flow convergence method and the jet area method for the determination of the severity of tricuspid regurgitation. METHODS AND RESULTS: The proximal isovelocity surface area radius and the jet area/length were measured in 71 consecutive patients with angiographically graded (grade 0/I-III) tricuspid regurgitation. Rank correlation coefficients with the angiographic grade were 0.71 (P < 0.001) for the proximal isovelocity surface area radius (aliasing border of 28 cm.s-1), 0.66 (P < 0.001) for the jet area, and 0.63 (P < 0.001) for the jet length. The proximal isovelocity surface area radius was significantly correlated with the jet area/length (correlation coefficients 0.82/0.77, P < 0.001). Correct differentiation between mild to moderate (grade I-II) and severe (grade III) tricuspid regurgitation was achieved in 62 of 71 patients (87%) by means of the proximal isovelocity surface area radius, in 61 of 71 (86%) by the jet area, and in 62 of 71 (87%) by the jet length. Grade III tricuspid regurgitation was not identified in five of 21 patients (24%) by means of the proximal isovelocity surface area radius, in six of 21 (29%) by the jet area, and in seven of 21 (33%) by the jet length. CONCLUSION: The flow convergence method and the jet area method are of similar value for the determination of the severity of tricuspid regurgitation. Both methods differentiated mild to moderate from severe tricuspid regurgitation in most patients. However, underestimation of severe tricuspid regurgitation in 20-30% of the cases represents a serious limitation of both methods.

Adult↗
PubMedDate not supplied

Estimates of maximum limits of food colours use in Brazil through the Danish Budget Method and the Bär and Würtzen-modified method.

The establishment of the permissible levels for the use of additives in foods must be based on the Acceptable Daily Intake (ADI). A method that may be applied for this purpose is the Danish Budget Method which estimates the maximum amount of the additive that may be added to the food based on the functional properties of the additive, and on the categories of the food in which the additive will be used. Based on the latest information Bär and Würtzen propose some modifications to the original Budget Method, one of which is the addition of a correction factor which takes into account the competition between different food additives with the same functional properties. In the present paper, both the Budget Method and the Bär and Würtzen-modified method were applied to evaluate whether the maximum levels of food colours use exceeded their ADI or not. Applying the original Budget Method, the results showed that the colours Sunset Yellow, Amaranth, Erythrosine, Ponceau 4R and Cochineal possibly exceeded the ADI; while applying the modified method only the colours Erythrosine and Cochineal would exceed the ADI. Brazilian regulatory authorities should be advised to establish maximum limits of use for the following categories of colours: Caramel, Inorganic, Natural and Artificial Colours Identical to the Natural Ones, where ADIs have been evaluated by JECFA.

Brazil↗
PubMed1978

Comparison of the conventional lecture method and the self-instruction method for teaching clinical pathology.

Two separate courses in clinical pathology were presented to two groups of second-year medical students. The first (large) group was taught by the conventional lecture method; the other (small) group, by a self-instructional method for roughly half of the course, followed by the conventional lecture method for the second half of the course. The effectivenesses of the two methods were compared, based on performances on the final written examination. It was found that the self-instructional method was as effective as the conventional method in teaching clinical pathology, at least in teaching basic principles and interpretive clinical chemistry, as judged by the performances of the two groups. Surprisingly, the two groups differed significantly in performances on the portion of the course taught by similar methods.

Education, Medical, Undergraduate↗
PubMed1998

Percutaneous dilational tracheostomy: the Ciaglia method versus the Portex [correction of Rapitrach] method.

UNLABELLED: In a prospective study, we performed percutaneous dilational tracheostomy (PDT) using either the Ciaglia method (gradual dilation) or the Portex [corrected] method (single dilation) in 80 patients. We encountered difficulty in dilating the tracheal stoma of three (7.5%) patients in the Ciaglia group because of tight pretracheal fascia. It was difficult to insert the tracheostomy tube in four (10%) patients in the Portex [corrected] group even after appropriate tracheal dilation. However, the tracheal cannulation was successfully completed in all patients in subsequent attempts. The mean time for completion of the procedures, from skin incision to insertion of the tracheostomy tube, was 14+/-5.5 min with the Ciaglia method and 6.5+/-3.5 min with the Portex [corrected] method. PDT with either method has not been associated with clinically significant hemorrhage, infection at the stoma site, or cosmetic deformity. In a follow-up period of 9 mo, none of our decannulated patients presented with clinical tracheal stenosis. Our results indicate that PDT with both methods is as safe and easy to organize and perform as a bedside procedure, obviating the need to transport critically ill patients from the critical care unit. IMPLICATIONS: The tracheas of 80 patients were cannulated through an artificial opening using either the Ciaglia (gradual dilation) or the Portex [corrected] (single dilation) method. Both techniques were successful with no significant complications. After 9 mo of closure of this opening, none of the survivors had significant scarring or narrowing of the trachea.

Adolescent↗
PubMed2000

Producing a full-scale model from computed tomographic data with the rapid prototyping technique using the binder jet method: a comparison with the laser lithography method using a dry skull.

Rapid prototyping using the binder jet method has recently been established and has already produced excellent results in industrial applications. The authors recently developed a technique for producing a full-scale model from computed tomographic (CT) data with the binder jet method as an approach to overcome the shortcomings of the laser lithography method, which is already widely used in medicine. They conducted a comparative investigation of full-scale models made with both techniques using a dry skull to determine the accuracy of the models. It was clearly demonstrated that the accuracy of the binder jet method was high enough to be used in craniomaxillofacial surgery because it was the same as the laser lithography method. This study employed data from the latest helical volume scan computed tomography device using a multidetector. The study showed that the new rapid prototyping technique was satisfactory in terms of speed, cost, installation environment, and accuracy of models, and that detailed shapes and structures can be reproduced well. Because this technique has many advantages over the laser lithography method, it should play a major role in craniomaxillofacial surgery and in other medical fields in combination with advances in CT devices. Although plaster is a more suitable fixation material when the emphasis is on the reproducibility of detailed structures, the binder jet method using starch is extremely useful for simulating operations and determining implant shapes because it allows for the prompt production of models.

Analog-Digital Conversion↗
PubMed1996

A comparison between a PCR method and a conventional culture method for detecting pathogenic Yersinia enterocolitica in food.

The aim of this study was to develop a polymerase chain reaction (PCR) method for the detection of pathogenic Yersinia enterocolitica and to compare it with an official culture method (NMKL-117). Primers were selected for nested PCR directed at the attachment invasion locus, ail, on the bacterial chromosome, as well as at a sequence on the pathogenic marker plasmid, termed virulence factor, virF. The final results obtained by the two methods were similar. However, while the conventional method yielded contradictory data for some steps the PCR method provided unambiguous results. Considerable advantages, i.e. higher sensitivity and specificity of the PCR method, compared with the conventional method for detecting pathogenic Y. enterocolitica, were demonstrated in this study.

Bacterial Outer Membrane Proteins↗
PubMed1996

Comparison of the Equil2 program and other methods for estimating the ion-activity product of urinary calcium oxalate: a new simplified method is proposed.

BACKGROUND: We aimed to develop a new simplified method to determine the urinary calcium-oxalate saturation. METHODS: The ion-activity product (AP) of urinary calcium oxalate was estimated in 345 clinical urine specimens (all 2.5 hour collections) using the Equil2 computer program and several other methods, and the results were compared. A new index using 4 parameters was created using multiple regression analysis. RESULTS: Our new simplified method (the AP(CaOx) index EQ2) provided a better approximation to the Equil2 method (r = 0.9897) than other conventional methods. CONCLUSION: This new method requiring 4 factors (urinary calcium, magnesium, oxalate, and citrate concentrations) is simple and clinically usable.

Calcium Oxalate↗
PubMed1996

A regression method including chronological and bone age for predicting final height in Turner's syndrome, with a comparison of existing methods.

A total of 235 measurement points of 57 Dutch women with Turner's syndrome (TS), including women with spontaneous menarche and oestrogen treatment, served to develop a new Turner-specific final height (FH) prediction method (PTS). Analogous to the Tanner and Whitehouse mark 2 method (TW) for normal children, smoothed regression coefficients are tabulated for PTS for height (H), chronological age (CA) and bone age (BA), both TW RUS and Greulich and Pyle (GP). Comparison between all methods on 40 measurement points of 21 Danish TS women showed small mean prediction errors (predicted minus observed FH) and corresponding standard deviation (ESD) of both PTSRUS and PTSGP, in particular at the "younger" ages. Comparison between existing methods on the Dutch data indicated a tendency to overpredict FH. Before the CA of 9 years the mean prediction errors of the Bayley and Pinneau and TW methods were markedly higher compared with the other methods. Overall, the simplest methods--projected height (PAH) and its modification (mPAH)--were remarkably good at most ages. Although the validity of PTSRUS and PTSGP remains to be tested below the age of 6 years, both gave small mean prediction errors and a high accuracy. FH prediction in TS is important in the consideration of growth-promoting therapy or in the evaluation of its effects.

Adolescent↗