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Biomedical subjects

Yuzuru Hayashi

Publications and source records attributed to Yuzuru Hayashi.

At least 19 recordsLinked to original sources

Prediction of measurement uncertainty in isotope dilution gas chromatography/mass spectrometry.

An equation is theoretically derived which describes the relative standard deviation (RSD) of the amount ratios of analyte to its isotope-labeled variant in gas chromatography/mass spectrometry (GC/MS) using the stable isotope dilution method. The determination of methyltestosterone is taken as an example. The uncertainty equation proposed is justified by comparing the theoretical RSD values with the experimental RSD values obtained by replication over a wide range of analyte amount. The detection limit and quantitation limit are estimated from the continuous plot (precision profile) of the theoretical RSD against analyte amount.

Deuterium↗

Method for estimating infection route and speed of influenza.

This paper puts forward a method for estimating the infection route and speed of influenza from the daily variations in the amount of influenza formulations supplied at distant city pharmacies. The cross-correlation function between the time variations at the pharmacies indicates as for the drug sales, how many days a pharmacy lags behind another pharmacy. The comparison of the time lags between the pharmacies can lead to the estimation of the infection route of influenza. Taking into account the distance between the locations of the pharmacies, we can calculate the infection speed of influenza. Three pharmacies located in Tokyo and its vicinity (Saitama and Kanagawa) are taken as an example. The thrust of this paper is to introduce the new strategy that can take full advantage of the information every pharmacy has in possession.

Antiviral Agents↗

Method for detecting health injury from prescriptions at a pharmacy.

A method is proposed to detect the abnormal situations of people's health in case of the unexpected outbreak of a disease by monitoring the daily variations in the prescriptions at a pharmacy. The abnormal situations are defined as the situations which are not included in the majority (99.9%) of the normal situations. An epidemic probably caused by infectious micro-organisms in a terrorist attack is taken as an example. The drugs for the typical symptoms are monitored: influenza anti-viral agent and common cold drug. This paper demonstrates that the border between the normal and abnormal situations corresponds to the detection limit which is a fundamental concept in analytical chemistry.

Antiviral Agents↗

Method for estimating the order of influenza infection between adults and children.

This paper puts forward a method for determining the infection order of influenza between adults and children from the daily variations in the amount of influenza anti-viral agents treated at a pharmacy. The time series of Tamiflu Capsule for adults and Tamiflu Dry Syrup for children are compared by means of the cross-correlation function. The results from pharmacies located in Tokyo and Kanagawa show that the influenza infection period of adults is earlier than that of children, indicating the infection order: first adults and second children. However, a pharmacy in Saitama yields no clear result.

Acetamides↗

An expression of uncertainty in calibration using stepwise or separate dilution of a stock solution.

The traditional method for linear calibration can estimate the confidence intervals of calibration lines from a set of experimental data for a single calibration line. However, the following situations, often encountered in laboratories, are out of reach of the method, since the concentrations of the standard solutions are not independent of each other: (A) a standard solution is diluted from a more concentrated one in a stepwise way (stepwise dilution); (B) every standard solution for a calibration experiment is prepared from a stock solution, but the stock solution is newly prepared for each calibration (separate dilution with the variable concentration of the stock solution). This paper puts forward a theory to calculate the confidence intervals of calibration lines in the above situations. Analyses made up of sample weighing, dilution, HPLC measurement and calibration with the linear least-squares fitting are taken as examples. The proposed theory is numerically compared to the traditional method.

Algorithms↗

[Estimation of detection limit from slope of B/B0 in competitive ELISA].

A method has been proposed for determining the detection limit (L(D)) from the slope of a semilogarithmic plot of a B/B(0) curve in competitive enzyme linked immunosorbent assay (ELISA). As an application, this paper describes a graphic determination of L(D) from analogue data in the literature. The L(D) obtained corresponds to the concentration at which the relative standard deviation of concentration estimates is 30%.

Calibration↗

Detection limit estimated from slope of calibration curve: an application to competitive ELISA.

This paper theoretically derives a general rule that while the slope of the semi-logarithmic plot (Y vs. log X) of a calibration curve varies depending on analyte concentration, X, the slope takes a specific value at the detection limit (L(D)). This rule holds good irrespective of the shape of the calibration curve (linear or non-linear) and in this paper, is applied to competitive ELISA (enzyme linked immunosorbent assay). The following relationship is deduced: slope of log-dose B/B0 at L(D) = [relative standard deviation (RSD) of blank responses] / 0.13. The L(D) obtained from the above-mentioned slope corresponds to the dose at which the RSD of dose estimates is 0.3 (= 30%). A commercial kit for 17alpha-hydroxyprogesterone is taken as an example.

17-alpha-Hydroxyprogesterone↗

New approaches to mechanism analysis for drug discovery using DNA microarray data combined with KeyMolnet.

We have developed a comprehensive information platform, named KeyMolnet, for drug discovery and life science research in the post-genome era. Using KeyMolnet, we show new approaches to research into the biological mechanism in DNA microarray analysis. Thanks to the DNA microarray technology, it is now possible to obtain very large quantities of gene expression data at a time. However, it is still difficult to extract meaningful information from such large quantities of data and to analyze the relationship between gene expression data and biological function. We therefore developed an advanced tool that can generate molecular networks upon demand, and beyond signaling "cross-talks," can connect them to physiological phenomena and medical and drug information. Here we show the methods of mechanism analysis using the DNA microarray data and KeyMolnet, as well as the possible mechanism of inducing apoptosis in the human promyelocytic leukemia cell line, HL-60, treated with 12-O-tetradecanoylphorbol 13-acetate (TPA), using the time series of gene expression data from DNA microarray experiments. KeyMolnet enables practical approaches to research into biological mechanisms, which in turn contribute to new discoveries in the medical, pharmaceutical and life sciences.

Apoptosis↗

Precision and detection limit of quality test for amorphous drug in powder X-ray diffractometry.

This report puts forward a method of powder X-ray diffractometry to estimate the precision and detection limit of the crystalline component in an amorphous drug. Cefditoren pivoxil (CP) was employed as a model drug. The major error source of the measurement at low crystal contents is shown to be the random noise in a diffraction pattern (halo pattern) of the amorphous material. For the analysis of the noise, the obstructive halo pattern should be eliminated from the observed pattern. The subtraction of the observed halo pattern from another one derived from the same material, extracts the random noise alone, although the noise is amplified by square root 2 times. The noise in the powder X-ray diffractometry was identified as the white noise. On the basis of the stochastic properties of the extracted noise and signal parameters (peak area) of CP, the relative standard deviations (R.S.D.) of the area measurements of the crystalline diffraction peaks were estimated over a wide range of crystal contents without repeated experiments. The detection limit was determined such that the crystal content at detection limit produced 30% R.S.D. of the measurements. The R.S.D. and detection limit obtained from FUMI theory were in good agreement with the results from the repeated measurements.

Anti-Bacterial Agents↗

An expression of within-plate uncertainty in sandwich ELISA.

This paper puts forward a method to describe an equation of the within-plate uncertainty (relative standard deviation (R.S.D.) of measurements) as a function of analyte concentration in sandwich enzyme-linked immunosorbent assay (ELISA). A kit for thyroid stimulating hormone is taken as an example. The pipetting procedures of analyte solution and chromogen-substrate solution and absorbance inherent to the wells of a microplate are identified as major error sources and their variability is included as parameters in the uncertainty equation. These parameters can be determined by the experiments with distilled water. The theoretical R.S.D. is shown to be in good agreement with the results of the repeated experiments using the real samples. Since the theory gives a continuous plot of R.S.D. against concentration, the uncertainty structure of the ELISA kit can be recognized over a wide concentration range and the detection limit and quantitation range can easily be determined on the plot.

Enzyme-Linked Immunosorbent Assay↗

Risk assessment of di(2-ethylhexyl)phthalate released from PVC blood circuits during hemodialysis and pump-oxygenation therapy.

This study deals with in vitro investigation of the release of di(2-ethylhexyl)phthalate (DEHP) during hemodialysis and pump-oxygenation therapy using medical grade PVC tubing. High resolution GC-MS analysis showed that the release of DEHP was time-dependently increased by circulation of bovine blood into a major system for the hemodialysis that is used in Japan, and the amount of DEHP released into the blood had reached 7.3 mg by 4 h of circulation. No significant difference was observed in the release patterns of DEHP under the conditions with and without fluid removal treatment during hemodialysis, indicating that the treatment seems not to be effective for eliminating DEHP from the blood through the hemodialysis membrane. Mono(2-ethylhexyl)phthalate (MEHP) analysis revealed that a small amount of DEHP (3-4%) was converted to MEHP by hydrolysis during the circulation of blood. A considerable amount of DEHP was also released from the PVC circuit mimicking the pump-oxygenation system, and 7.5-12.1 mg of DEHP had migrated into bovine blood from the circuit by 6 h. It was noticed, however, that the release was obviously suppressed by covalently coating the inner surface of the PVC tubing with heparin, though this effect was not observed with ionic bond type-heparin coating. Covalent bond type-heparin coating of PVC tubing seems to offer the advantage of decreasing the amount of DEHP exposure to patients during treatment using a PVC circuit.

Adult↗

Precision, limit of detection and range of quantitation in competitive ELISA.

This paper develops a mathematical model for describing the within-plate variation as the RSD (relative standard deviation) of absorbance measurements in a wide concentration range in competitive ELISA and proposes a method for determining the limit of detection (LOD) and range of quantitation (ROQ). The ELISA for 17 alpha-hydroxyprogesterone is taken as an example. The theoretical RSD description involves analyte concentration as an independent variable and error sources as parameters which concern the pipetting and absorbance measurement. Our model can dispense with repeated experiments of real samples, but the error parameters should be determined experimentally. The theory is in good agreement with the experiments. The most influential error sources at low and high sample concentrations are shown to be the pipetting of a viscous solution of antiserum and the absorbance inherent to the wells of a plate, respectively. The LOD and ROQ are defined as the concentration with 30% RSD and the region with <10% RSD, respectively, and are found in the theoretical plot of the RSD of concentration estimates vs concentration.

17-alpha-Hydroxyprogesterone↗

Improvement in reliability of probabilistic test of significant differences in GeneChip experiments.

A probabilistic test (FUMI theory) for GeneChip experiments has been proposed for selecting the genes which show significant differences in the gene expression levels between a single pair of treatment and control. This paper describes that the reliability of the judgment by the FUMI theory can be enhanced, when the selected genes are referred to biomolecular-functional networks of a commercial database. The genes judged as being differently expressed are grouped into a cluster in the biomolecular networks. It is also demonstrated that false positive genes have a trend in the networks to be isolated from each other, and also away from the clustered genes, since the false positive genes are randomly selected.

Algorithms↗

[Status of NIHS Computer Network System (NIHS-NET)].

We described the development of National Institute of Health Sciences Computer Network System (NIHS-NET), which was named NIHS Information and Computing Infrastructure (NICI) previously. In the system, the main server machines and common machines were replaced and the network lines were upgraded from 100 Mbps to 1Gbps. The connection nodes were changed from Inter Ministry Network (IMnet) to Science Information Network (SINET), and the dedicated lines between NIHS (yoga, osaka, tsukuba) and SINET were constructed. The Internet connection speed from each campus to SINET was upgraded. We also performed security audit in this system.

Computer Communication Networks↗

Estimation of uncertainty in kinetic-colorimetric assay of bacterial endotoxins.

The relative standard deviation (R.S.D.) of measurements is estimated in the kinetic-colorimetric assay of bacterial endotoxins without recourse to the usual repeated experiments. The measurements are the slopes of kinetic curves and two major factors are considered to cause the uncertainty of the measurements: (1) the pipetting of the sample and color development reagent; and (2) noise in the detection unit. The measurement R.S.D. is formulated as a function of endotoxin concentration. Two parameters (S.D. of the pipetted volumes and S.D. of the detector noise) are also required in the uncertainty equation, but no arbitrary coefficients are included. Since the S.D. values for pipettes and detector noise can be determined independently of the endotoxin assays, the measurement R.S.D. can be estimated by the above equation without repeating the assays. However, the calibration curve is necessary. The theoretical estimation is shown to be in good agreement with the experimental R.S.D. (n = 12) over a wide concentration range.

Algorithms↗

Prediction of measurement precision of apparatus using a chemometric tool in electrochemical detection of high-performance liquid chromatography.

The relative standard deviation (RSD) of measurements in high-performance liquid chromatography with electrochemical detection (HPLC-ECD) was predicted by a chemometric tool based on the 1/f fluctuation model which is made up of white noise and a Markov process, called the Function of Mutual Information (FUMI) theory. FUMI theory can provide aprecise and reliable detection limit from a single measurement of noise and signal in HPLC-ECD. To obtain RSD (n = 5) for determination of (-)-epicatechin at five concentrations required 12.5 h, while the predicted RSD by FUMI theory required only 0.5 h (one measurement). Moreover, to trace the source of instrumental noise, power spectra of chromatographic baseline were used. Selection of a suitable apparatus in HPLC-ECD system, acquisition of RSD, and detection limits for determination of catechins by HPLC-ECD were simply and easily made by this chemometric tool within a very short time. The use of the FUMI theory for the prediction of measuring precision was more efficient and the optimization was less time-consuming to be suited for determination.

Catechin↗

[Precision of internal standard method in HPLC analysis].

The internal standard methods are known to compensate for the errors from sample preparation and injection into an analytical instrument. However, recent HPLC apparatuses have injectors of excellent repeatability and it is dubious whether the cancellation of injection error can lead to substantial improvement in the precision of analysis. This paper answers the above question experimentally and theoretically. The HPLC analysis of butylscopolamine bromide is taken as an example. The relative standard deviations (RSD) of measurements in the internal standard method and absolute calibration curve method are compared and the advantages of these methods are discussed. The measurement RSD is shown to be well estimated by the (function of mutual information) (FUMI) theory without repeating measurements. This report also demonstrates simple equations for calculating the measurement RSD at an arbitrary concentration of analyte and for selecting the better method between the internal standard method and absolute calibration curve method under specific experimental conditions.

Calibration↗

Test of significant differences with a priori probability in microarray experiments.

A traditional method for comparing two expression levels of genes in microarray experiments is the two-sample t-test. Because of the difficulty in using a large number of microarrays, an alternative method is required which can provide a reliable judgment of the comparison from a small number of replicates, even from a single pair of control and treatment. We present a method for detecting the changes in the gene expression levels under two different conditions in microarray experiments. Our method targets a single experiment for each condition, while retaining the statistical advantages of the t-test. The new proposals are: 1) standard deviation (SD) estimates of the expression levels which are an indicator for significant differences are given a priori as a function of the expression levels; 2) the limit of detection (LOD) for the expression levels is used to eliminate the majority of genes expressed at extremely low levels. The a priori SD estimates are obtained from six replicates under a fixed condition and are shown to be the approximate, but proper description of the expression uncertainty covering diverse conditions (e.g., different samples (human and rat) and different DNA chips). The LOD is defined as three times blank SD according to the IUPAC recommendation. A cell line (HL60) which will undergo macrophage differentiation on treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA) is taken as an example. Our method is compared with the t-test for the data on duplicate TPA experiments and the former alone is evaluated with the data on a single TPA experiment. The errors from sample preparation and instrumental analysis are discussed.

Animals↗