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Electron microscopical demonstration of horseradish peroxidase by use of tetramethylbenzidine as chromogen and sodium tungstate as stabilizer (TMB-ST method): a tracing method with high sensitivity and well preserved ultrastructural tissue.

Until now methods using tetramethylbenzidine (TMB) for electron microscopy (TMB-EM methods) are all unable to provide a maximum demonstration of transported horseradish peroxidase (HRP) while maintaining good ultrastructural tissue preservation. In order to solve this problem, we have attempted to adapt a newly developed, highly sensitive TMB method using sodium tungstate (ST) as the stabilizer (TMB-ST method) for HRP electron microscopic retrograde and anterograde fiber tracing. The present study shows that the TMB-ST method combined with diaminobenzidine-cobalt (DAB-Co) is more sensitive than existing TMB-EM methods and that ultrastructural details are well preserved with this combined method. The resultant reaction product complex after osmication is stable and is observed as characteristic crystal-like structures which are extremely electron dense and often aggregated into clumps. In contrast, the TMB-ST method without the DAB-Co step frequently produces a moderate electron-dense reaction product. Therefore, we recommend the TMB-ST method combined with DAB-Co for HRP electron microscopy.

3,3'-Diaminobenzidine↗

Drug susceptibility testing in tuberculosis: a comparison of the proportion methods using Lowenstein-Jensen, Middlebrook 7H10 and 7H11 agar media and a radiometric method.

In the present investigation, the conventional drug susceptibility testing procedures using the 1% proportion method on Löwenstein-Jensen (LJ) and Middlebrook 7H10/7H11 agar media were compared with a recent radiometric method. The latter using "Bactec 460-TB" apparatus and a special broth medium (Becton and Dickinson Diagnostic Instrument Systems, Towson, MD, USA) gave drug susceptibility results within a week as compared to 4-5 weeks with the LJ method, and 3-4 weeks using 7H10/7H11 agar media. This report describes the critical concentrations retained for each of the methods and the methodology used. Also, the compatibility of the results obtained with the radiometric method with those obtained with conventional 7H11 agar method was perfect. In this report, we justify the choice of the 7H11 agar method instead of the LJ method as our conventional drug susceptibility procedure, and suggest also that radiometric drug susceptibility determination by Bactec 460-TB apparatus may be the method of choice for routine, rapid drug susceptibility determinations.

Colony Count, Microbial↗

Comparison of standard methods and gas chromatography method in determination of formaldehyde emission from MDF bonded with formaldehyde-based resins.

Formaldehyde emissions from MDF bonded with urea-formaldehyde resin (UF), melamine-formaldehyde resin (MF) and the co-polycondensation resin of urea-melamine-formaldehyde (UMF) and melamine-formaldehyde, measured by the Japanese standard method of determining formaldehyde emission with a desiccator (JIS A 5908) and the DIN EN 120 (European Committee For Standardization, 1991) method using the perforator value, were used as the typical standard methods. While the UF resin showed a desiccator value of 7.05 ppm and a perforator value of 12.1 mg/100 g panel, the MF resin exhibited a desiccator value of 0.6 ppm and a perforator value of 2.88 mg/100 g panel. According to the Japanese industrial standard and the European standard, the formaldehyde emission level of the MDF panels made with UF resin in this study was E(2) grade. The formaldehyde emission level was dramatically reduced by the addition of MF resin. This is because the addition of formaldehyde to melamine occurs more easily and completely than its addition to urea, even though the condensation reaction of melamine with formaldehyde is similar to that between urea and formaldehyde. These two methods, the desiccator method and the perforator method, produced proportionally equivalent results. Gas chromatography, a more sensitive and advanced method, was also used. The samples used for gas chromatography were gathered during the experiment involving the perforator method. The formaldehyde emission levels obtained from gas chromatography were similar to those obtained from the perforator method. The formaldehyde contents measured by gas chromatography were directly proportional to the perforator values.

Chemistry Techniques, Analytical↗

Development and evaluation of an in vitro method for prediction of human drug absorption II. Demonstration of the method suitability.

The aim of the present work was to assess the actual suitability and general applicability of a new in vitro permeation method based on an appositely developed artificial membrane to evaluate and predict drug absorption potential. The proposed method was employed to evaluate the apparent permeability of a set of 21 structurally diverse drugs having very different solubility and permeability properties, covering the whole range of fraction absorbed in humans (F(a) from 13 to 100%); 13 of the drugs in this study were part of the list suggested by FDA for validation of in vitro permeation methods. An excellent linear correlation (R(2)=0.957) was obtained between artificial membrane apparent permeability and human absorption data in the whole range of F(a) values examined (including all the drugs belonging to the above FDA list), indicating the good predictive ability of the proposed method not only for highly absorbed hydrophobic compounds but also, differently from other in vitro permeation methods, for poorly or middling permeable drugs. The predictive ability of the new method was greater than those obtained for the same set of drugs with PAMPA and Caco-2 permeability literature data, probably due to the poor sensitivity of these methods towards hydrophilic drugs. The better performance of our artificial membrane was attributed to the hydrophilic nature of the support that, differently from the commonly used hydrophobic supports, offers less resistance to permeation of hydrophilic compounds. A comparison of permeation data of theophylline, ketoprofen, aciclovir and furosemide (selected, respectively, as models of I-IV BCS classes) obtained using a Caco-2 cell based dynamic method and the developed artificial membrane and the corresponding F(a) values in humans further confirmed the suitability of the proposed permeation method as predictor of the oral absorption of passively absorbed drugs.

Acyclovir↗

Development of a harmonised method for the profiling of amphetamines V: Determination of the variability of the optimised method.

This paper is the fifth in a series of six in relation to the development of a harmonised method for the profiling of amphetamine [L. Aalberg, K. Andersson, C. Bertler, H. Borén, M.D. Cole, J. Dahlén, Y. Finnon, H. Huizer, K. Jalava, E. Kaa, E. Lock, A. Lopes, A. Poortman-van der Meer, E. Sippola, Development of a harmonised method for the profiling of amphetamines I. Synthesis of standards and compilation of analytical data, Forensic Sci. Int. 149 (2005) 219-229; L. Aalberg, K. Andersson, C. Bertler, M.D. Cole, Y. Finnon, H. Huizer, K. Jalava, E. Kaa, E. Lock, A. Lopes, A. Poortman-van der Meer, E. Sippola, J. Dahlén, Development of a harmonised method for the profiling of amphetamines II. Stability of impurities in organic solvents, Forensic Sci. Int. 149 (2005) 231-241]. The third paper [K. Andersson, K. Jalava, E. Lock, L. Aalberg, Y. Finnon, H. Huizer, E. Kaa, A. Lopes, A. Poortman-van der Meer, M.D. Cole, J. Dahlén, E. Sippola, Development of a harmonised method for the profiling of amphetamines III. Development of the gas chromatographic method, Forensic Sci. Int., in press] dealt with the optimisation of the gas chromatographic and detection methods whereas the fourth paper [K. Andersson, K. Jalava, E. Lock, Y. Finnon, S. Stevenson, L. Aalberg, H. Huizer, E. Kaa, A. Lopes, A. Poortman-van der Meer, M.D. Cole, J. Dahlén, E. Sippola, Development of a harmonised method for the profiling of amphetamines IV. Optimisation of sample preparation, Forensic Sci. Int., in press] concerned the optimisation of the extraction method prior to GC analysis. This paper is a study of the optimised method in order to determine its stability. Investigations of within and between day variations were carried out in four laboratories. Moreover, variations between laboratories were also determined. Both flame ionisation detector (FID) and MS detection were used. One laboratory studied nitrogen-phosphorous detector (NPD) detection as well. For this task, 12 batches of amphetamine were prepared. Six of them were synthesised via the Leuckart route, three via the nitrostyrene route and three via the reductive amination route [A.M.A. Verweij, Impurities in illicit drug preparations: amphetamine and methamphetamine, Forensic Sci. Rev. 1 (1989) 2-11]. Taking into account all studied target compounds and the average results from four laboratories, the within day variation was around 6% for FID and 5% for MS, the between days variation was around 10% for FID and 8% for MS. For NPD detection, within day variation was 5% and between days variation 9% (only one laboratory). Finally, the inter-laboratory variation was about 12% for FID (four laboratories) and 10% for MS (three laboratories).

Journal Article↗

Merits of HPLC-based method over spectrophotometric method for assessing the kinetics and inhibition of mammalian adenosine deaminase.

Measurement of adenosine deaminase (ADA) activity using spectrophotometric method presents problem, regarding the quantitative estimation of the substrate degradation and product formation, due to the closely apposed lambda(max) of the substrates, product and the inhibitor. The feasibility of applying reverse-phase HPLC technique, for studying adenosine deaminase-catalyzed reaction product and inhibition study was examined. We have drawn a comparison between the HPLC-based method over the corresponding spectrophotometric method. A gradient elution pattern was used to separate substrate (adenosine and deoxyadenosine), product (inosine and deoxyinosine) and standard adenosine deaminase inhibitor (erythro-9-(3-nonyl-p-aminobenzyl)-adenine) in the HPLC method. The product formation was quantitated by monitoring the absorbance at 260 nm with the progress of time. The limit of detection as well as the limit of quantification of the respective enzymatic product were found to be in nano molar (nM) range in the HPLC method. This study was also extended to monitor adenosine deaminase activity in different cancer cells of hematological origin. The HPLC-based method is found to be suitable for the quantitative estimation of adenosine deaminase-catalyzed reaction product and for studying inhibition mechanism of different inhibitors. The HPLC-based method has specific advantages over the spectrophotometric method. Moreover, the concentration of different nucleotides in cell lysate and body fluid can be measured using this HPLC method.

Adenosine Deaminase↗

Preparation of human frozen-thawed seminal specimens using the SpermPrep filtration method: improvements over the conventional swim-up method.

OBJECTIVE: To examine the qualitative and quantitative characteristics of frozen-thawed spermatozoa recovered through the SpermPrep (ZBL, Inc., Lexington, KY) filtration method or the swim-up technique for the purpose of intrauterine insemination (IUI) or other techniques for assisted reproduction. DESIGN, SETTING, PATIENTS: Thirty pairs of frozen specimens purchased from three commercial semen suppliers were used in this study. Each pair consisted of two aliquots from the same semen specimen. MAIN OUTCOME MEASURES: Spermatozoa recovered via the SpermPrep filtration and swim-up processes were evaluated for sperm numbers recovered, sperm motility, grade of sperm motility, percentage of morphologically normal spermatozoa, the response of spermatozoa to a hypo-osmotic environment (hypo-osmotic swelling test), and the amount of debris present. RESULTS: Application of the SpermPrep filtration method resulted in recovery of significantly greater numbers of spermatozoa (P less than 0.01) than were recovered with the swim-up method (31.1 +/- 3.2 x 10(6) versus 10.2 +/- 1.8 x 10(6) spermatozoa, respectively). This represents a mean recovery of approximately one half (49%) of all spermatozoa applied to the filter, whereas for the swim-up method, it was only 15%. The overall quality of recovered spermatozoa was virtually identical between the two methods (P greater than 0.05). The percent motile sperm improved by a mean of 18% to 20%, the grade of motility improved by a mean of 0.4 points (scale 0 to 4), the percent of morphologically normal spermatozoa increased by a mean of approximately 10%, the percent of spermatozoa reactive to a hypo-osmotic medium test increased by a mean of approximately 9%, and the debris score decreased by a mean of 0.2 to 0.3 points (scale 0 to 4). Most importantly, the mean total number of motile, morphologically normal spermatozoa after filtration through the SpermPrep column was 20.2 +/- 1.1 x 10(6), representing a mean recovery of 73% of the normal spermatozoa originally applied to the column. This was 316% greater than the yield obtained with the swim-up method (6.4 +/- 0.8 x 10(6)), which was significantly greater (P less than 0.01) than that recovered via the swim-up method. Also, the time required to harvest sperm through SpermPrep filtration was 20 to 25 minutes versus 80 minutes required for the swim-up method (P less than 0.05). CONCLUSION: Considering that the effectiveness of frozen-thawed semen is already limited when compared with fresh semen, SpermPrep filtration is the method of choice over the swim-up technique of sperm selection because the former provides significantly greater numbers of high quality sperm. It should be considered as an adjunct in semen preparation for IUI or other forms of assisted reproduction.

Evaluation Studies as Topic↗

Comparison of two stereological methods for quantitative renal morphology: a modified fractionator and modified Weibel-Gomez method.

To determine the best method for quantitative analysis of renal structure, we compared two stereological techniques: the unbiased fractionator method and the model-based method described by Weibel-Gomez. Kidneys of 20 week old pregnant female Sprague Dawley rats were investigated. Using these two stereological techniques, the mean glomerular volume and the total glomerular number per kidney were determined in 16 kidneys of 8 rats. Both methods were used in a modified way to correct for variations in section thickness (fractionator) and to reduce the length of the measuring process (Weibel-Gomez), respectively. The mean glomerular volume was comparable in both methods (Fractionator: 5.49 +/- 0.56 x 10(4) mm3, Weibel-Gomez: 5.35 +/- 0.34 x 10(4) mm3). In contrast, using the Weibel-Gomez method (43,774 +/- 2338), the total number of glomeruli per kidney was significantly higher than that obtained by the fractionator technique (39,359 +/- 4250). The results as well as the time necessary for each method and the practicability of the techniques were compared. In our hands, apart from the respective advantages and disadvantages, the Weibel-Gomez technique is easier to perform and much more efficient than the probably more elegant fractionator method. The bias problem of the Weibel-Gomez method does not play an important role with respect to the most common biological problems.

Animals↗

Dissolution testing of acetylsalicylic acid by a channel flow method-correlation to USP basket and intrinsic dissolution methods.

A new modification of the channel flow dissolution method is introduced together with the theoretical basis to extract the solubility and mass transfer parameters from the dissolution experiments. Correlation of drug dissolution profiles in the channel flow apparatus was evaluated with respect to USP basket and intrinsic dissolution methods at pH 1.2 or 6.8. Acetylsalicylic acid (ASA) was studied as a pure drug substance and as three simple tablet compositions with microcrystalline cellulose (MCC) and/or lactose as excipients. The channel flow measurements of 100% ASA tablets correlated well with the results of intrinsic dissolution tests. In the channel flow method as well as in the USP basket method the release of ASA was fastest from the tablet compositions containing lactose, while the slowest dissolution rate was observed with the composition containing MCC as the only excipient. As presumed, the dissolution rate of the weak acid was decreased as the pH of the medium was lowered, which was clearly confirmed also by the three dissolution methods. MCC forms matrix tablets and in the USP basket method the dissolution profiles followed square root of time kinetics indicating that diffusion was the rate-controlling step of ASA dissolution. Also the channel flow results indicated that the dissolution of ASA was controlled by mass transfer. The swelling behaviour of the tablets is different in the channel flow method as compared to the basket method: only one tablet surface is exposed to the dissolution medium in the channel flow system. The contact between the tablet surface and the dissolution medium is more similar between the channel flow and intrinsic dissolution methods.

Algorithms↗

A method for the quantitative analysis of iron speciation in meat by using a combination of spectrophotometric methods and high-performance liquid chromatography coupled to sector field inductively coupled plasma mass spectrometry.

The determination of the heme and non-heme iron fractions in raw and cooked beef steak by using spectrophotometric methods and high-performance liquid chromatography coupled to a double-focusing sector field inductively coupled plasma mass spectrometer (HPLC-SF-ICPMS) is reported. Size exclusion chromatography coupled to SF-ICPMS was used to measure the iron-containing biomolecules in the samples. This approach allowed for the direct on-line detection of the most abundant iron isotope 56Fe without interference from 40Ar16O. The HPLC-ICPMS results for the iron speciation analysis of a raw beef steak, used as an analytical quality control (AQC) sample, showed that the main iron biomolecule present was the heme iron-containing protein myoglobin. For the AQC sample, the agreement among the HPLC-ICPMS method, the non-heme iron spectrophotometric method, and the total iron concentration showed 100% recovery of iron. The sum of the different iron-containing compounds determined using the developed HPLC-ICPMS method accounted for all the iron-containing compounds extracted. The analysis of myoglobin in steak by HPLC-ICPMS showed that on cooking the concentration was reduced by 85%. However, a spectrophotometric method specific for heme iron showed that it was still intact, even after heating to 80 degrees C. The measurement of the total iron in the cooked steak and the HPLC extracts by inductively coupled plasma optical emission spectroscopy (ICP-OES) indicated that the extraction method for the HPLC analysis was no longer applicable and that loss of the heme group from the protein rendered it incompatible with the size exclusion separation. The detection limit (concentration equivalent to 3 times the baseline for a blank injection) of the HPLC-ICPMS method was 2.4 ng as iron. The results demonstrate that a combination of analytical methods can be used to provide valuable information about dietary levels of nutritionally important metal-containing compounds as well as the efficiency of established extraction methods for raw and cooked meat samples.

Animals↗

Assessment of experimental bond dissociation energies using composite ab initio methods and evaluation of the performances of density functional methods in the calculation of bond dissociation energies.

Composite ab initio CBS-Q and G3 methods were used to calculate the bond dissociation energies (BDEs) of over 200 compounds listed in CRC Handbook of Chemistry and Physics (2002 ed.). It was found that these two methods agree with each other excellently in the calculation of BDEs, and they can predict BDEs within 10 kJ/mol of the experimental values. Using these two methods, it was found that among the examined compounds 161 experimental BDEs are valid because the standard deviation between the experimental and theoretical values for them is only 8.6 kJ/mol. Nevertheless, 40 BDEs listed in the Handbook may be highly inaccurate as the experimental and theoretical values for them differ by over 20 kJ/mol. Furthermore, 11 BDEs listed in the Handbook may be seriously flawed as the experimental and theoretical values for them differ by over 40 kJ/mol. Using the 161 cautiously validated experimental BDEs, we then assessed the performances of the standard density functional (DFT) methods including B3LYP, B3P86, B3PW91, and BH&HLYP in the calculation of BDEs. It was found that the BH&HLYP method performed poorly for the BDE calculations. B3LYP, B3P86, and B3PW91, however, performed reasonably well for the calculation of BDEs with standard deviations of about 12.1-18.0 kJ/mol. Nonetheless, all the DFT methods underestimated the BDEs by 4-17 kJ/mol in average. Sometimes, the underestimation by the DFT methods could be as high as 40-60 kJ/mol. Therefore, the DFT methods were more reliable for relative BDE calculations than for absolute BDE calculations. Finally, it was observed that the basis set effects on the BDEs calculated by the DFT methods were usually small except for the heteroatom-hydrogen BDEs.

Journal Article↗

Non-invasive assessment of cardiac output during exercise in healthy young humans: comparison between Modelflow method and Doppler echocardiography method.

AIMS: The Modelflow method can estimate cardiac output from arterial blood pressure waveforms using a three-element model of aortic input impedance (aortic characteristic impedance, arterial compliance, and systemic vascular resistance). We tested the reliability of a non-invasive cardiac output estimation during submaximal exercise using the Modelflow method from finger arterial pressure waveforms collected by Portapres in healthy young humans. METHODS: The Doppler echocardiography method was used as a reference method. Sixteen healthy young subjects (nine males and seven females) performed a multi-stage cycle ergometer exercise at an intensity corresponding to 70, 90, 110 and 130% of their individual ventilatory threshold for 2 min each. The simultaneous estimation of cardiac output (15 s averaged data) using the Modelflow and Doppler echocardiography methods was performed at rest and during exercise. RESULTS AND CONCLUSION: The Modelflow-estimated cardiac output correlated significantly with the simultaneous estimates by the Doppler method in all subjects (r = 0.87, P < 0.0001) and the SE of estimation was 1.93 L min-1. Correlation coefficients in each subject ranged from 0.91 to 0.98. Although the Modelflow method overestimated cardiac output, the errors between two estimates were not significantly different among the exercise levels. These results suggest that the Modelflow method using Portapres could provide a reliable estimation of the relative change in cardiac output non-invasively and continuously during submaximal exercise in healthy young humans, at least in terms of the relative changes in cardiac output.

Adult↗

New hybrid method for reactive systems from integrating molecular orbital or molecular mechanics methods with analytical potential energy surfaces.

A computational approach to calculating potential energy surfaces for reactive systems is presented and tested. This hybrid approach is based on integrated methods where calculations for a small model system are performed by using analytical potential energy surfaces, and for the real system by using molecular orbital or molecular mechanics methods. The method is tested on a hydrogen abstraction reaction by using the variational transition-state theory with multidimensional tunneling corrections. The agreement between the calculated and experimental information depends on the quality of the method chosen for the real system. When the real system is treated by accurate quantum mechanics methods, the rate constants are in excellent agreement with the experimental measurements over a wide temperature range. When the real system is treated by molecular mechanics methods, the results are still good, which is very encouraging since molecular mechanics itself is not at all capable of describing this reactive system. Since no experimental information or additional fits are required to apply this method, it can be used to improve the accuracy of molecular orbital methods or to extend the molecular mechanics method to treat any reactive system with the single constraint of the availability of an analytical potential energy surface that describes the model system.

Journal Article↗

Comparison of three homogeneous methods for measuring high-density lipoprotein cholesterol with a precipitation method in diabetic and non-diabetic subjects.

Three homogenous methods for the automated measurement of high-density lipoprotein-cholesterol (HDL-C) are studied and compared with a precipitation method in diabetic and non-diabetic subjects. All three methods meet the precision criteria of the National Cholesterol Education Programme (NCEP). Triglycerides did not cause significant interference up to 10 mmol/L in the immuno-inhibition method (Wako) and up to 40 mmol/L in the Randox and Roche methods. Haemoglobin, up to a concentration of 5 g/L, had only negligible effect on the performance of all three homogeneous methods. Bilirubin caused an increasing positive bias in all methods above a concentration of 50 micromol/L. In comparison with the precipitation method, the new homogeneous methods agreed for type 1 diabetic patients but showed a positive bias for the control subjects and patients with type 2 diabetes. The bias of HDL-C levels in type 2 diabetes may be sufficient to affect the calculation of cardiovascular risk, and may therefore influence the decision to prescribe lipid-lowering medication.

Bilirubin↗

Comparison of six respirator fit-test methods with an actual measurement of exposure in a simulated health care environment: Part II--Method comparison testing.

This article, the second in a series of three, describes the method comparison testing portion of a study conducted to compare the fit factors from six quantitative fit-tests (QNFT) with a measure of a respirator wearer's actual exposure assessed by end-exhaled air analysis for 1,1,2-trichloro-1,2,2-trifluoroethane (Freon-113) under the same conditions. The six QNFT methods were (1) continuous low flow, flush probe; (2) continuous high flow, deep probe (CHD); (3) exhalation valve discharge (EVD); (4) controlled negative pressure; (5) 10-minute Ambient Aerosol 1 (AA1); and (6) 30-minute Ambient Aerosol 2. The first three methods utilized corn oil and a forward light scattering photometer. The last two methods used the TSI Portacount. Respirators used in the study were both disposable and elastomeric organic vapor/high efficiency half-masks. The characterization equations from the preliminary research (described previously) were used to determine the actual exposure to Freon-113 during the method comparison testing. The fit factors resulting from the QNFT methods were then individually correlated with the Freon-113 exposures using the coefficient of determination, R2. The lowest R2 value, 0.20, was found with the EVD method. The highest R2 values, 0.81 and 0.78, were associated, respectively, with the CHD and AA1 methods. This study suggests that some QNFT methods may be used to estimate actual respirator performance under laboratory conditions.

Adult↗

A new scanning method for thallium-201 myocardial SPECT: semidecubital position method.

The authors have developed a new scanning method for Tl-201 myocardial SPECT using a rotating gamma camera with 180-degree data collection. This requires a new type of imaging table on which the patient is placed in a 50-degree right semidecubital position to allow close proximity to the camera-collimator system. Using this method, attenuation by the imaging table is completely avoidable because the projected views in the left oblique directions are not obstructed by the table. In addition, the radius of camera rotation is about 25% shorter than in the conventional method. Phantom and 16 clinical studies were performed to compare the performance of conventional and semidecubital position methods. In phantom studies, tomographic images produced using the new method showed better lesion contrast and more counts in the posteroinferior wall of the heart than those produced using the conventional method. Clinically, 12 of the 16 studies showed more counts in the posteroinferior wall of the heart with the semidecubital position method than with the conventional method. The semidecubital position method is useful for Tl-201 myocardial SPECT, especially for evaluation of posteroinferior wall hypoperfusion.

Gamma Cameras↗

Method to resolve microphone and sample location errors in the two-microphone duct measurement method

Utilizing the two-microphone impedance tube method, the normal incidence acoustic absorption and acoustic impedance can be measured for a given sample. This method relies on the measured transfer function between two microphones, and the knowledge of their precise location relative to each other and the sample material. In this article, a method is proposed to accurately determine these locations. A third sensor is added at the end of the tube to simplify the measurement. First, a justification and investigation of the method is presented. Second, reference terminations are measured to evaluate the accuracy of the apparatus. Finally, comparisons are made between the new method and current methods for determining these distances and the variations are discussed. From this, conclusions are drawn with regards to the applicability and need for the new method and under which circumstances it is applicable. Results show that the method provides a reliable determination of both microphone locations, which is not possible using the current techniques. Errors due to inaccurate determinination of these parameters between methods were on the order of 3% for R and 12% for Re Z.

Journal Article↗

Krylov subspace iterative methods for boundary element method based near-field acoustic holography.

The reconstruction of the acoustic field for general surfaces is obtained from the solution of a matrix system that results from a boundary integral equation discretized using boundary element methods. The solution to the resultant matrix system is obtained using iterative regularization methods that counteract the effect of noise on the measurements. These methods will not require the calculation of the singular value decomposition, which can be expensive when the matrix system is considerably large. Krylov subspace methods are iterative methods that have the phenomena known as "semi-convergence," i.e., the optimal regularization solution is obtained after a few iterations. If the iteration is not stopped, the method converges to a solution that generally is totally corrupted by errors on the measurements. For these methods the number of iterations play the role of the regularization parameter. We will focus our attention to the study of the regularizing properties from the Krylov subspace methods like conjugate gradients, least squares QR and the recently proposed Hybrid method. A discussion and comparison of the available stopping rules will be included. A vibrating plate is considered as an example to validate our results.

Journal Article↗