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[Quantitative analysis of cerebrospinal fluid dynamics in syringomyelia using cine MRI with pre-saturation].

Some reports show the qualitative analysis of cerebrospinal fluid (CSF) pulsation in the subarachnoid space and the syrinx using cine magnetic resonance imaging (MRI). However, few reports studied the quantitative analysis of CSF pulsation. We report here the results of quantitative analysis of CSF pulsation using the cine MRI with pre-saturation pulse. Using flow phantom, we calibrated the correlation between the true velocity and calculated velocity acquired from the movement of pre-saturated low signal. Three cases of syringomyelia with Chiari malformation, two cases of traumatic syringomyelia, and three normal volunteers were examined using this technique, and we could obtain time-velocity curves of CSF in both subarachnoid spaces and syrinx. Although obvious pulsation of CSF was observed in the syrinx of all Chiari malformations, no pulsation of CSF was observed in the syrinx of traumatic syringomyelia. CSF in the syrinx moves upward in the early systolic cardiac cycle, downward in the systolic cycle, and upward again in the diastolic cycle. Patterns of CSF pulsation in syrinx were similar to those in the subarachnoid space, but "phase" of these pulsations differed from case to case. These results suggest the close relationship between CSF pulsation in the syrinx and CSF obstruction at the cranio-vertebral junction. Dissociation of the "phase" of CSF pulsation in the syrinx and subarachnoid space may be significant in the analysis of the pathogenesis of syringomyelia.

Adult↗

Quantitative analysis using steady-state free precession nuclear magnetic resonance

The use of steady-state free precession nuclear magnetic resonance (NMR) for quantitative analysis in low magnetic field is investigated and shown to exhibit substantial advantages compared to more conventional NMR methods. With only minor additional requirements, the technique permits a considerable increase in signal-to-noise ratio for a given acquisition time. The experimental conditions needed for implementation and optimization of the acquisition parameters are explored and shown to be easily accessible with unsophisticated equipment. The applicability of the technique for quantitative analysis of samples with a range of relaxation rates is tested using various examples of practical interest. Highly reproducible results can be obtained much faster and without any special sample-dependent adjustments.

Journal Article↗

[Quantitative analysis of myocardial tracer distribution in patients with ischemic heart disease: comparison of 201T1 and 123I-15-(p-iodophenyl)-3-methylpentadecanoic acid (BMIPP)].

Quantitative assessment of myocardial tracer uptake in stress-delayed thallium and resting BMIPP imagings were performed in 24 patients with coronary artery disease. Each distribution was displayed on the bull's eye polar map and % uptake of each distribution was calculated as a mean value in 9 myocardial segments on the polar map. Redistribution index (% delayed uptake minus % stress uptake on thallium images) and discordance index (% delayed thallium uptake minus % BMIPP uptake) were also calculated. Each parameter was compared to the visual uptake score and wall motion score on contract left ventriculography. Excellent correlations were obtained between % uptake and the uptake score in each tracer. The % thallium and BMIPP uptake also correlated with regional wall motion score. Furthermore, a significant correlation was observed between redistribution and discordance indexes in the mildly hypoperfused segments. These data indicate that the quantitative analysis of thallium and BMIPP distributions seems to be valuable to understand relationship between perfusion and regional wall motion. The discordant BMIPP uptake may represent asynergic but viable segments. However, several important factors, such as attenuation factor should be also taken into consideration for such quantitative analysis.

Aged↗

Indirect UV detection as a non-selective detection method in the qualitative and quantitative analysis of heparin fragments by high-performance capillary electrophoresis.

The application of capillary electrophoresis (CE) in combination with indirect UV detection for the qualitative and quantitative analysis of synthetic low-molecular-mass heparin fragments, at low pH, is described. It is demonstrated that, in contrast to direct UV detection, with indirect UV detection the signal obtained for various synthetic heparin pentasaccharides is nearly independent of their molecular structure. Moreover, the sensitivity of indirect UV detection is at least one order of magnitude higher than that of direct UV detection. CE-indirect UV detection for the qualitative and quantitative analysis of low-molecular-mass glycosaminoglycans was achieved by using 5 mM 5-sulphosalisylic acid, pH 3 or 5 mM 1,2,4-tricarboxybenzoic acid, pH 3.5 as electrophoresis buffer and chromophore. The technique is exemplified by the analysis of three pharmaceutical preparations of synthetic heparin pentasaccharides. The method employing indirect UV detection was validated with respect to repeatability, limit of detection, limit of quantitation, linearity, accuracy and ruggedness. In the indirect detection mode, the limit of detection for synthetic pentasaccharides is below 5 fmol, whereas the limit of quantitation is about 25 fmol. The method shows excellent repeatability and is linear in the femtomole-picomole range. Finally, it is demonstrated that the method is suitable for the analysis of various types of glycosaminoglycans.

Carbohydrate Sequence↗

Quantitative analysis of regional left ventricular function after myocardial infarction in the pig assessed with cine magnetic resonance imaging.

To assess the accuracy of quantitative analysis of global and regional wall motion and wall thickening of the left ventricle with cine magnetic resonance (MR), images obtained in eight pigs before and after myocardial infarction were compared with those obtained using gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA)-enhanced multislice spin-echo MR imaging and determination of pathology. The region with abnormal wall motion and wall thickening, as determined with cine MR imaging, identified the same region of infarction as indicated by Gd-DTPA-enhanced spin-echo MR imaging and pathology. Within the infarcted region wall motion and wall thickening analyzed with the centerline method were significantly reduced. We conclude that the use of quantitative analysis of cine MR images accurately determines localization and extent of regional left ventricular dysfunction in the infarcted heart in vivo. This analysis using dedicated software including the centerline method allows sequential assessment of regional left ventricular function in normal and infarcted hearts.

Animals↗

Temporal and Quantitative Analysis of Chimerism in Liver and Kidney Transplant Patients: An Enhanced Fluorescence Detection System Allows for More Sensitive Quantitative Detection.

Background: Because spontaneous microchimerism has been reported in stable renal and hepatic allografts, the presence of donor-derived cells in recipient tissues was investigated in kidney and liver tranplant recipients. Methods and Results: Human lymphocyte antigen class II markers and Y-chromosome sequences in male donor-to-female recipient transplants were used for chimeric analysis. Human lymphocyte antigen typing was performed by group-specific polymerase chain reaction amplification and restriction fragment length polymorphism analysis, X-chromosome- and Y-chromosome-specific primers were used in a multiplex polymerase chain reaction analysis. Quantitative Y-chromosome analysis was performed using energy-transfer fluorescence from a nested primer system. Patients who had rejected their grafts were also analyzed, as were a group who were analyzed for chimerism at the time of transplant (day 1) and sequentially at various intervals for up to 3 months. Of 23 long-term kidney patients analyzed, 16 were chimeric by human lymphocyte antigen or sex-determination analysis. In 2 patients whose graft had failed no chimerism was observed. Chimerism in liver patients was detectable on the day of transplant and was maintained for 30 to 120 days as measured at 5-day intervals (these patients continue to be monitored). Quantititative analysis suggested that the ratio of donor to recipient cells was variable in a patient and ranged from greater than 1 in 10(4) to less than 1 in 10(5). An enhanced fluorescence energy-transfer detection system was adopted to increase sensitivity of the polymerase chain reaction detection of chimerism and to quantitate the results. Conclusions: The results indicate that cells from the donor organ migrate into recipient tissues early after transplantation. These cells persist in a majority of patients for at least 3 to 4 years. It has been proposed that tolerance is related to the presence of these "passenger" leukocytes and that dendritic cells play the most important role. The data suggest that the establishment of chimerism plays an important role in graft acceptance in a majoritiy of the kidney and liver patients in this study. These findings also suggest that the levels of chimeric cells, "a quantitative chimerism," may be important in establishing tolerance but further studies are required to support this contention.

Journal Article↗

Semi-quantitative analysis of indigo by surface enhanced resonance Raman spectroscopy (SERRS) using silver colloids.

In this paper we report for the first time semi-quantitative analysis of indigo using surface enhanced Raman spectroscopy (SERS) and surface enhance resonance Raman spectroscopy (SERRS). Indigo, a dye widely used today in the textile industry, has been used, historically, both as a dye and as a pigment; the latter in both paintings and in printed material. The molecule is uncharged and largely insoluble in most solvents. The application of SERS/SERRS to the semi-quantitative analysis of indigo has been examined using aggregated citrate-reduced silver colloids with appropriate modifications to experimental protocols to both obtain and maximise SERRS signal intensities. Good linear correlations are observed for the dependence of the intensities of the SERRS band at 1151 cm(-1) using laser exciting wavelengths of 514.5 nm (R=0.9985) and 632.8 nm (R=0.9963) on the indigo concentration over the range 10(-7)-10(-5) and 10(-8)-10(-5) mol dm(-3), respectively. Band intensities were normalised against an internal standard (silver sol band at 243 cm(-1)). Resonance Raman spectra (RRS) of aqueous solutions of indigo could not be collected because of its low solubility and the presence of strong fluorescence. It was, however, possible to obtain RS and RRS spectra of the solid at each laser excitation wavelength. The limits of detection (L.O.D.) of indigo by SERS and SERRS using 514.5 and 632.8 nm were 9 ppm at both exciting wavelengths. Signal enhancement by SERS and SERRS was highly pH dependent due to the formation of singly protonated and possibly doubly protonated forms of the molecule at acidic pH. The SERS and SERRS data provide evidence to suggest that an excess of monolayer coverage of the dye at the surface of silver colloids is observed at concentrations greater than 7.85x10(-6) mol dm(-3) for each exciting wavelength. The data reported herein also strongly suggest the presence of multiple species of the indigo molecule.

Colloids↗

Quantitative analysis of inorganic particulate burden in situ in tissue sections.

Quantitative information on tissue particulate burden is essential for investigation of relationships of particulates to several aspects of disease, such as exposure, physiology, radiology and pathology. We have developed a method for quantitative analysis of inorganic particulate burden in situ in tissue sections using scanning electron microscopy (SEM) with backscattered electron (BSE) imaging and energy dispersive x-ray analysis (EDXA) or the ion microprobe mass analyzer (IMMA). Our data show reproducible results for concentrations above 10(6) particles per cm3 of tissue. Samples of less than one microgram can be analyzed. Standard 5 microns paraffin sections of formalin fixed tissue are mounted on carbon discs, deparaffinized and examined in the SEM under standardized conditions. Fields of view comprising 10,000 microns3 are searched and all particles visible in the BSE image are counted and individually analyzed by EDXA. Location, number, size and types of particles are tabulated. Previous work showed correlation of quantitative SEM/EDXA with bulk analysis for Ni (r = 0.64), and IMMA with bulk analysis for Be (r = 0.999). Analytical results on exogenous and endogenous particles in 40 lungs (greater than 6000 fields searched and greater than 5700 particles analyzed), and the pathology observed in the tissue, may serve as baseline for future studies on all types of inorganic particles in normal and abnormal lungs. Observed concentrations ranged from 3 X 10(6) to 2 X 10(9) exogenous particles per cm3 of tissue.

Asbestosis↗

[Quantitative analysis of mRNA expression of the LDL receptor and HMG-CoA reductase in bovine tissue with competitive RT-PCR].

The present study exemplifies the method of quantitative reverse transcriptase-polymerase chain reaction (cRT-PCR) by quantifying, in bovine tissues, the specific mRNAs for the two key proteins which regulate cellular cholesterol metabolism, the low density lipoprotein (LDL)-receptor and the 3-hydroxy-3-methylglutaryl-Coenzyme A (HMG-CoA) reductase. Our data reveal a broad range of expression for both mRNAs. The LDL-receptor mRNA expression was highest in the adrenal gland (6.7 x 10(4) molecules/micrograms cellular RNA) followed by the corpus luteum of the ovary, intermediate in the liver (8.6 x 10(2) molecules/micrograms cellular RNA) and not detectable in the large intestine. Expression of the HMG-CoA reductase almost paralleled LDL-receptor values. It was found to be highest in the ovary (6.1 x 10(6) molecules/micrograms cellular RNA) followed by the adrenal gland, intermediate in the liver (3.1 x 10(5) molecules/micrograms cellular RNA) and lowest in the large intestine (9.9 x 10(4) molecules/micrograms cellular RNA). These data are consistent with other reports concerning the quantitative analysis of the expression of both proteins as determined by other experimental approaches. Thus cRT-PCR is a valuable tool for the quantitative analysis of the LDL-receptor and the HMG-CoA reductase.

Adrenal Glands↗

Quantitative analysis of polymerase chain reaction products by dot blot.

Quantitative analysis of polymerase chain reaction (PCR) products is usually accomplished by gel electrophoresis and Southern blotting. We have developed an alternative technique that allows PCR products to be directly quantitated from unfractionated samples. The PCR was used to amplify genomic (endogenous) DNA sequences (actin) and exogenous DNA (herpes simplex virus-1 (HSV-1) ribonucleotide reductase) isolated from the trigeminal ganglia of rabbits to demonstrate the dot blot method of PCR product analysis. Two primer pairs (actin and ribonucleotide reductase) were coamplified, resulting in two different PCR products. Duplicate aliquots of the PCR products were applied to separate nylon membranes and hybridized with 32P-labeled oligonucleotide probes. Each radioactive probe was specific for target (HSV-1 DNA) or control (actin DNA) products. Quantitation using a laser scanning PhosphorImager and ImageQuant software demonstrated that the dot blot method can be used to rapidly analyze a large number of PCR samples.

Animals↗

Quantitative analysis of hydroxyl radicals in the anterior optic nerve of the cat following transient ischemia.

PURPOSE: Ischemic insults in the optic nerve are relatively common ocular disorders. This study provides quantitative data on the generation of hydroxyl radicals in the anterior optic nerve of the cat during the early reperfusion phase following 90 minutes of ischemia. MATERIALS AND METHODS: Cats were injected intravenously with sodium salicylate that by acting as an hydroxyl radical trap, forms 2,3-dihydroxybenzoic acid (2,3-DHBA). Ischemia was achieved by intraocular pressure (IOP) elevation via cannulation of the anterior chamber. Ischemia was maintained for 60 minutes in 6 eyes and 90 minutes in 12 eyes, followed by 5 minutes of reperfusion. In an additional 6 eyes, ischemia was maintained for 90 minutes without reperfusion. RESULTS: After 90 minutes of ischemia and reperfusion, mean normalized levels of 2,3-DHBA (ie, ng 2,3-DHBA/microg salicylate/mg protein) that represent the levels of hydroxyl radicals were 2.47 times (at least) higher in the anterior optic nerves than in the fellow sham-operated controls (P = 0.03). These levels were 3.9 times (at least) greater than in the corresponding levels after 90 minutes of ischemia without reperfusion (P= 0.005). CONCLUSION: This study provides a quantitative analysis and evidence for the generation of hydroxyl radicals in the optic nerve of the cat following transient ischemia. A quantitative analysis may provide an important tool to detect even relatively small levels of free radicals in the tissues, and to evaluate the relative efficacy of various therapeutic agents in the inhibition of free radical generation following ischemia and reperfusion.

Animals↗

Quantitative analysis of polymorphic mixtures of ranitidine hydrochloride by Raman spectroscopy and principal components analysis.

Ranitidine hydrochloride exists as two polymorphs, forms I and II, both of which are used to manufacture commercial tablets. Raman spectroscopy can be used to differentiate the two forms but univariate methods of quantitative analysis of one polymorph as an impurity in the other lack sensitivity. We have applied principal components analysis (PCA) of Raman spectra to binary mixtures of the two polymorphs and to binary mixtures prepared by adding one polymorph to powdered tablets of the other. Based on absorption measurements of seven spectral regions, it was found that >97% of the spectral variation was accounted for by three principal components. Quantitative calibration models generated by multiple linear regression predicted a detection limit and quantitation limit for either forms I or II in mixtures of the two of 0.6 and 1.8%, respectively. This study demonstrates that PCA of Raman spectroscopic data provides a sensitive method for the quantitative analysis of polymorphic impurities of drugs in commercial tablets with a quantitation limit of less than 2%.

Drug Evaluation, Preclinical↗

Quantitative analysis of the two macrophage colony-stimulating factor mRNA expressed in a human stromal cell line by reverse transcription-polymerase chain reaction (RT-PCR).

We established a quantitative analysis system for 4.0 kb and 1.6 kb macrophage colony-stimulating factor (M-CSF) mRNA, using reverse transcription-polymerase chain reaction. Using this system, we performed quantitative analysis of the two mRNAs expressed in the human stromal cell line, KM102, in the resting condition and when stimulated by various concentrations of interferon-gamma (IFN-gamma). The expression of 1.6 kb M-CSF mRNA was more efficiently stimulated by IFN-gamma than that of 4.0 kb M-CSF mRNA. The alternative splicing of a single M-CSF gene has been shown to generate several M-CSF proteins with different localization; we believe that molecular analysis of the transcription products by this system is important to better understand the physiological significance of the different species of M-CSF derived from each mRNA.

Base Sequence↗

[Qualitative and semi-quantitative analysis of local wall motion abnormalities by gated-blood-pool in comparison to biplane cineventriculography (author's transl)].

Left ventricular wall motion at rest was assessed by the following three methods in 37 patients of whom 12 patients had coronary artery lesions of > 75% and Ecg-signs of transmural MI: A) qualitative analysis of GBP, B) semi-quantitative analysis of GBP, C) qualitative analysis of a biplane cineventriculogram. Sensitivity and specificity of each method were evaluated based on the cineventriculographic findings. Specificity of method A was 83%, of method B 100%. Sensitivity of method A was 88%, of method B 84%. Concordance of methods B and C was 92.8% (167/180 left ventricular segments). Discordance between the two methods was found only in 1 segment judged to be akinetic and in 12 segments judged to be hypokinetic by one of the two methods.

Adult↗

[A quantitative analysis of the phrenic nerve activities during the cough reflex].

The phrenic nerve activity is an appropriate indicator for the output of the respiratory center as a whole, although a quantitative analysis of each of the frequency band components that constitute the neural activities during the respiratory reflex has not yet been made by other investigators. In the present study, we have made a quantitative analysis of each of the frequency band components of phrenic nerve activity during the cough reflex in anesthetized dogs. The efferent activities of the phrenic nerve were recorded from the central cut end of the phrenic nerve. Each fraction of phrenic nerve activity was separated into bands spanning a range of 100 Hz each by a variable filter and analyzed using a program for the power spectrum. The cough reflex was induced by mechanical stimulation of the tracheal mucosa. In the power spectrum analysis of each frequency band, increase in the power of each of the frequency band components was observed during the cough reflex. Particularly, the power of the 2 approximately 100 Hz band components increased significant as compared with the other frequency band components. Such as increase in the power of the 2 approximately 100 Hz band components toward the other frequency band components during the cough reflex was not observed after an i.v. administration of DMPP and lobeline, but the power increased extended over every frequency band. These findings suggest that the process for integrating the cough reflex was different from that of the other respiratory reflexes and that the present method is useful for investigating the central mechanisms of the cough reflex.

Action Potentials↗

Semi-quantitative analysis of indigo carmine, using silver colloids, by surface enhanced resonance Raman spectroscopy (SERRS).

The application of surface enhanced resonance Raman spectroscopy (SERRS) to the semi-quantitative analysis of the dye, indigo carmine, has been examined using citrate-reduced silver colloids. Good linear correlations are observed for the dye band at 1580 cm(-1) in the concentration range 10(-7)-10(-5) and 10(-9)-10(-5) mol dm(-3), using laser exciting wavelengths of 514.5 [(R=0.9983)] and 632.8 nm [(R=0.9978)], respectively. At concentrations of dye above 10(-6) M the concentration dependence of the SERRS signals is non-linear due to the coverage of the surface of the colloidal particles by the dye being in excess of a full monolayer. At concentrations above 10(-6) M resonance Raman spectroscopy (RRS) can be employed for the quantitative analysis of the dye. An internal standard was used and a good linear correlation (R=0.997) was observed for the dependence of dye signal intensities at 1580 cm(-1) in the concentration range 10(-5)-10(-4) M using a laser exciting wavelength of 514.5 nm. The limits of detection of indigo carmine by SERRS (514.5 nm), SERRS (632.8 nm) and solution RRS (514.5 nm) are found to be 0.9, 1 and 38 ppm, respectively.

Colloids↗

[Quantitative analysis of smooth pursuit eye movement].

Abnormalities of smooth pursuit eye movement (SPEM) have been estimated, mainly using the wave form on an electro-oculogram, in a qualitative way. Many methods for quantitative analysis of SPEM have been designed, though most are still uncommon in present clinical use. Using a personal computer, we developed a method of automatic quantitative analysis of ocular tracking eye movement recorded by electro-oculography (EOG). The design concept of this method is based on the observation that eye movement during ocular tracking consists of two different kinds of eye movements, one is SPEM and the other is saccade. The combination of SPEM and saccade (composite eye movement: CEM) commonly appears during ocular tracking. These two kinds of eye movement are essentially different not only in behavior but also about involved neural pathway in the central nervous system. From this point of view, we believe that the two kinds of eye movements involved in ocular tracking should be evaluated separately. The analysis method is outlined as follows. A horizontal sinusoidally moving visual target was employed to elicit ocular tracking eye movements. The test frequencies were set at 0.1, 0.2, 0.4 and 0.8Hz, and the amplitude of target motion was 15 deg at each frequency. The 20 seconds of eye movement data measured by EOG were fed into the computer through a digital-analog converter for further analysis. Using our original saccade detection algorithm, based on the physiological behavior of saccades, the saccadic components were detected and removed from the eye movement wave. The remaining parts, fragments of SPEM, were connected by means of interpolating defective parts. The reconstructed wave was a slow cumulative eye position curve (SCEP). Sinusoidal target motion, CEM and SCEP were processed by the FFT (Fast Fourier Transformation) method. Bode plots were applied to summarize the gain and phase of responses to SCEP and the target motion wave. These processes enable us to estimate abnormalities of SPEM such as low gain, abnormal phase shift and large trends in tested duration. We conclude that the method described here is useful for quantitative estimation of SPEM in clinical neuro-otological examinations.

Adult↗