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PubMed1985

A potential microneurosurgical method for the manipulation of the developing nervous system of the postimplantation mouse embryo in utero. I. Injection methods.

A microsurgical method for the injection of cells into the postimplantation mouse embryo whilst it is in utero is described. Specific regions of the 9.5-day-old mouse embryo brain have been injected with marker substances, such regions including the intraventricular cavity of the telencephalon and the trigeminal facialis ganglion bulge of the forebrain. Histological study of the injected embryos shows that the injection method is precise and able to avoid damage to other nearly embryonic tissues. The method should have wide application in the study of questions of normal cell lineage, neoplasia, teratology, and toxicology.

Animals↗

PubMed1992

Comparison of a novel thin-layer chromatographic-fluorescence detection method with a spectrofluorometric method for the determination of 7-hydroxycoumarin in human urine.

A novel method for the determination of 7-hydroxycoumarin in human urine which combines thin-layer chromatography (TLC) with fluorescence detection (FD) has been devised. The limit of detection (1 ng/ml) enables determination of 7-hydroxycoumarin after both administration of coumarin and environmental exposure to this fragrance material. When compared to a spectrofluorometric method of analysis, the TLC-FD method proved to be more selective for the analysis of 7-hydroxycoumarin in human urine.

Administration, Oral↗

PubMed1994

Comparison of assay methods by second-derivative spectroscopy, colorimetry and fluorescence spectroscopy of salicylic acid in aspirin preparations with a high-performance liquid chromatographic method.

Second-derivative spectroscopy, colorimetry and fluorescence spectroscopy have been compared with a high-performance liquid chromatographic (HPLC) method for the assay of salicylic acid in preparations of aspirin. Results are presented for the linearity, sensitivity and reproducibility of these methods. The second-derivative spectroscopic and the HPLC methods were acceptable in terms of linearity, sensitivity and inter-day reproducibility and were convenient for the routine analysis of salicylic acid in aspirin preparations.

Aspirin↗

PubMed1985

Rapid methods in clinical microbiology. Clinical situations in which rapid methods may best be applied.

Utilization of rapid microbiologic methods could improve both the quality of patient care and help to reduce patient health care costs through more efficient patient management. Medically urgent situations such as meningitis, endocarditis, bacteremia, some soft tissue infections, ocular infections, and pneumonia in immunocompromised hosts are obvious areas for application of rapid methods. However, efficient management of outpatient infections such as group A streptococcal pharyngitis, bacteriuria, diarrhea, and sexually transmitted diseases could also benefit from rapid test results. Rapid methods for detection of slow growing or unculturable microorganisms represent a promising area for development of new test methodologies. Realization of the potential of various rapid microbiology tests, however, requires that several difficult questions be addressed. Acceptable test rapidity and accuracy must be established by concensus of microbiologists and clinicians. Moreover, new approaches to reduce total test turn-around time and novel means to facilitate physician use of rapid test results must be sought in the immediate future.

Bacterial Infections↗

PubMed1992

Methods of monitoring menstrual function in field studies: efficacy of methods.

Efficacy of methods for monitoring female reproductive potential under field study conditions was evaluated. Women (n = 10) were recruited to participate for two menstrual cycles on the bases, in part, of not seeking fertility assistance, working full-time but not in the medical field, and having less than one year of college education. Luteinizing hormone (LH), estrone-3-glucuronide, and pregnanediol-3-glucuronide were measured in daily morning urine and normalized to creatinine concentrations. These urinary measures were parallel to serum LH, estradiol, and progesterone profiles. Based on these urinary measures, 6 of 19 cycles were judged to be atypical. Transvaginal ultrasonography provided insights into ovarian activity during the atypical cycles. Of 13 LH surges detected by radioimmunoassay, 7 were not detected by a semiquantitative dipstick (OvuSTICK), perhaps due to that method's sensitivity to loss of LH immunoactivity caused by sample freezing. While intervals from salivary and vaginal mucous electrical resistance signals to the LH surge during typical cycles were similar to those reported previously, they were not predictive of ovulatory status during atypical cycles. Fifty-three percent of the cycles were misclassified on the basis of the basal body temperature rise. Cervical mucous color, amount, and consistency were not predictive of ovulation under these study conditions. The results from these 19 menstrual cycles provide information about the efficacy of various methods for characterizing menstrual function under field study conditions. In this regard, urinary endocrine measures are the most informative or practical.

Adult↗

PubMed2005

[Quantification of bladder-outlet obstruction in males: standard method vc VBN method].

GOAL: To compare the merits of two methods, standard (Abrams-Griffiths number, Schafer's classification, etc.) and VBN, to study bladder-outlet obstruction in men with benign prostatic hyperplasia (BPH). MATERIAL AND METHODS: The parameters deduced from both methods and from repeated pressure-flow studies were computed for a population of patients with BPH. Correlation coefficients between standard and VBN parameters were systematically evaluated. Test-retest reliability and inter-rater reliability of the VBN parameters were investigated. RESULTS: The VBN obstruction parameter was linearly corelated to the A-G number (R = 0.992) and thus is related to the provisional ICS obstruction nomogram. A simple modification to the standard index projected isometric pressure (PIP) yields an mPIP parameter strongly correlated with the VBN detrusor contraction-strength parameter (R = 0.962). VBN analysis reveals minor phenomena such as premature fading of the detrusor contraction to be responsible for much of the void-to-void variability of pressure-flow studies. Consequently, the primary VBN obstruction and contraction-strength parameters exhibit better test-retest and inter-rater reliability than the standard parameters and are less sensitive to changes in testing circumstances (bladder volume, urethral catheter size and psychological factors). CONCLUSION: With the standard approach to test bladder obstruction, two values: A-G and mPIP, derived from pressure-flow studies, best qualify the patient's voiding status. These parameters are simple to calculate but are sensitive to testing variations and minor phenomena. The VBN approach is more complicated, but its parameters for obstruction and detrusor contraction strength are less dependent on testing variations.

Aged↗

PubMed2005

Generic method for systematic phase selection and method development of biochromatographic processes. Part I. Selection of a suitable cation-exchanger for the purification of a pharmaceutical protein.

Even if the first protein therapeutics are now for more than 20 years on the market the selection of suitable adsorbents for the preparative downstream processing (DSP) of these biomolecules as well as the method development towards process conditions are still based mainly on 'trial and error'. Therefore, theses processes are not perfectly efficient, but indeed very time consuming and laborious. In this study a novel systematic method is introduced to find a suitable adsorbent (not necessarily the best one) with appropriate separation parameters for a specific separation with reduced effort. Following this strategy, the adsorbents must first be packed into columns under preparative conditions and then characterized completely with regard to, e.g. pressure drop, k'-values, plate heights (HETP curves), selectivity and capacity by using test substances, which are similar in their characteristics (molecular mass, size, charge distribution, hydrophobicity) to the target proteins. With the database once determined, a preselection of most suitable adsorbents including separation parameters is made regarding chromatographic and also economical properties. After this, preparative experiments must be conducted with a reduced number of adsorbents to figure out the individual influence of side components. This approach is demonstrated for the separation of an exemplary industrial protein mixture using cation-exchange chromatography (CEX). Characterization of different weak CEX-adsorbents is illustrated. After comparing these phases with each other, a first preselection and a prediction of suitable adsorbents is made. In the following preparative separation conditions (load, velocity, gradient) are determined for the preparative separations using the database and results of some additional experiments. The final comparison of separation performance in preparative scale confirms this selection and so the applicability of the new method.

Cation Exchange Resins↗

PubMed2005

Development of a solid-phase microextraction method for direct determination of pentachlorophenol in paper and board samples: Comparison with conventional extraction method.

A solid-phase microextraction (SPME) method has been developed for the determination of pentachlorophenol (PCP) in paper and board samples. The analytical procedure involves direct extraction of PCP from paper and board samples and determination by gas chromatography with electron capture detection (GC-ECD). Two kinds of commercially available fibres; 100 microm polydimethylsiloxane (PDMS), apolar, and 85 microm polyacrylate (PA), quite polar, were evaluated to determine the extraction efficiency of pentachlorophenol. Parameters affecting the extraction process, such as temperature and time, were studied. Moreover, time of desorption and the effect of addition of salt were also investigated. The optimized procedure was applied to the analysis of pentachlorophenol (PCP) in five samples of virgin and recycled paper and board. The PCP content was determined by GC-ECD. To evaluate the effectiveness of the proposed method, it was compared with conventional extraction method with liquid-liquid extraction and derivatization. Detection limit of 0.015 microg/g for PCP in paper was achieved with a RSD of 14%.

Chemical Fractionation↗

PubMed2004

Reference methods for the measurement of free thyroid hormones in blood: evaluation of potential reference methods for free thyroxine.

In this paper, a review of the literature on the methods for the detection of FT(4) is provided. Furthermore, an overview of the most important uncertainties and interferences of the potential reference methods for determination of free thyroxine (free triiodothyronine) are also presented. Especially, the separation step-dialysis, or ultrafiltration-implies important technical and theoretical uncertainties and interferences as to the methods.Furthermore, a short review of methods based on isotope dilution/mass spectrometry (ID/MS) to quantify thyroxine is provided. There has so far been no attempt to quantify the free fraction of thyroxine by ID/MS, probably due to lack of sensitivity, but if quantification of the free fraction of thyroxine can be done with ID/MS, then this step seems the least compromised step only including minor uncertainties in comparison to the separation step.

Calibration↗

PubMed2006

Susceptibility testing of Mycobacterium tuberculosis by broth microdilution method: a rapid alternative method.

A total of 50 clinical isolates of Mycobacterium tuberculosis were tested by the proportion method and the broth microdilution method (BMM) for primary antitubercular drugs. The results were obtained after 14 days of incubation. There was a 100% agreement for all drugs except streptomycin, which showed a 96% agreement. Thus, the BMM was found to be a rapid, reliable, and less labor-intensive method to determine MIC.

Antitubercular Agents↗

PubMed2006

Development of a harmonised method for the profiling of amphetamines VI: Evaluation of methods for comparison of amphetamine.

Amphetamine samples were analysed by gas chromatography-mass spectrometry (GC-MS), and the peak areas of 33 target compounds were transformed by applying various pretreatment techniques. The objective was to optimise the ability of a number of distance metrics to establish links between samples of amphetamine originating from the same batch (henceforth referred to as linked distances). Furthermore, partial least squares discriminant analysis (PLS-DA) was used to evaluate the effects of various pretreatment methods on separation of amphetamine batches synthesised by the Leuckart reaction, reductive amination of benzyl methyl ketone, and the nitrostyrene route. The most efficient way to pretreat GC-MS data varied for the different distance metrics, although best results were obtained when data were normalised to the sum of peak areas, and either the fourth root or a logarithm was applied to the normalised data. When pretreating normalised data by fourth root transformation, Pearson correlation was the distance metric that was most successful at finding linked samples. Normalisation and the use of fourth root also represented the best method of pretreating data when employing PLS-DA to separate samples synthesised by different routes. To achieve a faster and more user-friendly procedure for evaluating chromatograms, experiments were performed in which the number of target compounds used to compare samples was reduced. The effect of each compound that was removed was studied by applying PLS-DA and by using Pearson correlation to calculate linked distances as well as unlinked distances (between samples from different batches of amphetamine). Considering both links between samples from the same batch and separation of samples synthesised by different routes, the best results were obtained with the data set comprising 26 compounds. Finally, it was found that the profiling method developed in this work was superior to an existing technique with respect to separating linked and unlinked distances.

Journal Article↗

PubMed2004

Method for producing coded micro-carrier and test method by using a novel type biochip.

This paper provides a method for producing a novel type coded micro-carrier. A simple and cost effective solution for bio-molecule applications was developed. Application relevant items such as manufacture process, biospecific interaction, and analysis method are discussed. For low cost fabrication, the use of LIGA-like process is suggested. LIGA-like process is used as a dry patterning process in which an intense beam of light from an excimer laser is used to pattern a material directly. This process has found extensive application in the microelectronics industry for patterning of polymer materials. The use of LIGA-like techniques offers two attractive features: first, we can cut the polymer into many tiny micro-carriers with micrometer precision. Second, LIGA-like process allows to encode with high precision spatial information onto the micro-carrier that can be used in the identification of the bio-molecule. This paper gives a description of the basic idea, describes the fabrication of the novel micro-carrier that we called "coded micro-carrier," and of the image processing algorithms used for the analysis of bio-molecules. This study also provides a test method for identifying a bio-molecule, which includes mixing several coded micro-carriers with the hybridized unknown bio-molecules; and identifying the codes on the micro-carrier via image recognition system. The numbers and types of the known micro-carrier can be flexibly adjusted according to the number of tested bio-molecules.

Animals↗

PubMed1979

Application of a high-performance liquid chromatographic fluorescence method for the rapid determination of alpha-tocopherol in the plasma of cattle and pigs and its comparison with direct fluorescence and high-performance liquid chromatography-ultraviolet detection methods.

High-performance liquid chromatography is used to develop a sensitive method for the determination of tocopherol levels in the plasma of cattle and pigs. This method is compared with a similar method using UV detection and one using direct fluorescence determination of tocopherol. Finally a double injection technique used in conjunction with fluorescence detection is shown to enhance the rate of analysis of the tocopherol levels in bovine plasma extracts.

Animals↗

PubMed1989

Selection of high-performance liquid chromatographic methods in pharmaceutical analysis. III. Method validation.

The most important steps in the validation of high-performance liquid chromatographic (HPLC) methods are discussed. The establishment of system suitability data and the assessment of peak purity are demonstrated on the example of bisquaternary amino steroids. For the recognition of incomplete resolution of adjacent peak pairs, the absorbance-ratio method in which the ratio of absorbances at two preselected wavelengths are plotted as a function of time in combination with the separation of sample components subjected to various chemical and physico-chemical treatments (stress conditions) is applied. The separation power and performance of the HPLC systems are characterized by the system resolution (SR) and system selectivity (SS). The special demands of stability-indicating methods are summarized.

Androstane-3,17-diol↗

PubMed1997

Development of a supercritical fluid extraction method for determination of lipid classes and total fat in meats and its comparison with conventional methods.

A method has been developed for the determination of total fat and lipid classes in meat using supercritical fluid extraction (SFE). The results agreed well with results from the conventional Bligh and Dyer and Schmid, Bondzynski and Ratzlaff extraction methods. SFE has advantages compared to the latter methods of a low consumption of hazardous organic solvents and shorter extraction time. After investigation of several different conditions, the most rapid extraction was achieved by adding 1 ml of cyclohexane to a 0.5-g sample mixed with 1 g Hydromatrix in a 7-ml thimble. The optimized SFE parameters were: 370 bar, 50 degrees C, 8% ethanol modifier, 4 ml/min dynamic flow for 30 min and collection with a tube leading to a vial.

Chromatography, Gas↗

PubMed2000

Method development for the determination of anthocyanins in red wines by high-performance liquid chromatography and classification of German red wines by means of multivariate statistical methods.

A simple and fast HPLC method without sample pretreatment is described for the separation of anthocyanins in red wines using a new pH-stable stationary phase. The linearity between peak area and concentration and ruggedness of the method were checked. Investigations were made about the safekeeping of red wine samples concerning anthocyanins. Classification of 52 different wine samples was performed by multivariate statistical methods.

Anthocyanins↗

PubMed1991

A new method for the analysis of the dynamics of the molecular genetic control systems. II. Application of the method of generalized threshold models in the investigation of concrete genetic systems.

Mathematical models of the prokaryotic control systems of tryptophan biosynthesis (both normal and with cloned blocks) and arabinose catabolism have been built using the method of generalized threshold models. Kinetic curves for molecular components (mRNAs, proteins, metabolites) of the systems considered are obtained. It has been shown that the method of generalized threshold models gives a more detailed qualitative picture of the dynamics of the molecular genetic control systems in comparison with the heuristic method of threshold models. The qualitative analysis of the functioning of the following mechanisms of control of the tryptophan biosynthesis: (1) inhibition of the activity of anthranilate synthetase by tryptophan, (2) repression and (3) attenuation of transcription of the tryptophan operon on the basis of the mathematical model of the control system of the tryptophan biosynthesis demonstrates that feedback inhibition is the most operative of the considered mechanisms while repression allows the bacterium to economize intracellular resources. As regards the control system of the arabinose catabolism the results of modelling enable us to state the following. The induction by arabinose within a wide range of parameter values causes two subsystems (araBAD and transport operons) of the arabinose regulon with a low rate of arabinose utilization to pass into a stationary regime and one subsystem (araC operon) to pass into a stable periodical regime. A study of the system characterized by the effective utilization of arabinose has shown that under induction by arabinose stable oscillations with small amplitudes of the concentration of regulatory protein and oscillations with large amplitudes of the concentrations of arabinose-isomerase and transport protein may occur. The period of the oscillation depends on the mean lifetime of the "activator-DNA" complex and on the rate constant of arabinoseisomerase degradation.

Animals↗

PubMed1983

Enzyme cytochemical staining of individual cells with the use of a polyacrylamide carrier. Studies on the synthetizing reaction technique, the indigogenic method, the metal salt method, the post-azo-coupling technique, and the tetrazolium salt technique.

A recently developed method for the (quantitative) demonstration of glucose-6-phosphate dehydrogenase activity in individual cells with the use of a polyacrylamide carrier has been extended for other enzyme cytochemical techniques. Isolated hepatocytes have been incorporated in the matrix of a thin transparent polyacrylamide gel prior to incubation in a cytochemical medium. The techniques which have been applied are the synthetizing reaction technique for glycogen phosphorylase, the indigogenic method for nonspecific esterase, the metal salt method for glucose-6-phosphatase, the post-azo-coupling technique for acid phosphatase, and the tetrazolium salt technique for succinate and lactate dehydrogenase activities. In all cases a few major problems which occur in the cytochemistry on single cells seem to be solved. The morphology is very well preserved, the final reaction product seems to be precipitated at the expected site of enzyme activity and the coloured end-product is highly specific for the enzyme activity to be studied, as has been demonstrated well with control experiments. The conclusion is reached, therefore, that this relatively simple device can be used routinely for the optimalization of enzyme cytochemistry of single cells.

Acrylic Resins↗