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PubMed1990

Quantitative analysis of walking in patients with knee osteoarthritis: a method of assessing the effectiveness of non-steroidal anti-inflammatory treatment.

Most therapeutic tests of osteoarthritis treatments are assessed by criteria based either on fundamental data or on clinical data, which is often subjective. A quantified analysis of locomotion can be used to determine the spatiotemporal indices (stride length and duration), kinematic indices (walking speed, velocity peak), and symmetry criteria that are relevant to the assessment of locomotor handicaps in patients with osteoarthritis. This study examined the progression of locomotor abilities in 11 subjects aged 49-69 (mean 60.9) years with knee osteoarthritis before and after treatment with a non-steroidal anti-inflammatory drug. Naproxen sodium (1100 mg) was given once a day for seven days. The condition before and after treatment was evaluated by quantitative analysis of locomotion, estimation of pain on a visual analogue scale, and assessment of the degree of functional disability. Significant improvement in locomotor indices (proportional increase in walking speed 17.8%) and in degree of pain (proportional decrease 27%) as estimated on the analogue scale was found after non-steroidal anti-inflammatory drug treatment. The lack of a significant correlation between the decrease in pain experienced by the patients and the objective improvement of their functional capabilities emphasises the need in further studies of new treatments to accompany the patients' own assessments of self improvement with a quantitative analysis of the way in which they walk.

Aged↗
PubMed2003

A fast Western blot procedure improved for quantitative analysis by direct fluorescence labeling of primary antibodies.

The procedures for Western blots have been around for a long time and recent developments have increased the sensitivity for luminescent techniques so that the need for radioactive probes has been limited to only a few applications. Nevertheless, most protocols require more than 6 h and are often performed over more than a day. The majority of techniques require a secondary antibody conjugated to an enzyme that catalyzes a color reaction in order to amplify a detectable signal. However, both processes, the binding of a secondary antibody and the catalyzed reaction with the dye, are sources for errors and the latter is disadvantageous for a signal that is linear over a larger range of detected antigen. In order to improve the procedure most commonly used for quantitative analysis and convenience, we investigated the use of fluorescence labeling of primary monoclonal antibodies against Escherichia coli RNA polymerase subunits (beta', sigmaE and sigmaFecI) and their use in Western blots. We achieved a sensitivity (<1 ng detectable protein) comparable to most luminescent techniques. Additionally, we reduced the procedure time significantly to less than 1 h after SDS-PAGE and transfer to a membrane. Above all, we obtained a linear signal over the range of 30 ng to 1 microg of protein (dependent on protein size) making quantitative analysis of Western blots easier and more reliable.

Antibodies, Monoclonal↗
PubMed1996

Quantitative analysis of numerical chromosome aberrations in various morphological types of colorectal carcinomas.

Quantitative analysis by fluorescence in situ hybridization (FISH) on thin paraffin-embedded tissue sections, using specific probes for chromosomes 11, 17, and 18 was employed in various morphological types of early and advanced colorectal cancer to clarify tumor cytogenetics. The chromosome index (CI) was calculated as a quantitative measure of the chromosome copy number. Compared with the CI of normal epithelium, the CI of chromosome 11 in villous components of adenomas or polypoid early cancers was decreased, while the CI in flat type or advanced colorectal cancers, conversely, was increased (P < 0.05). The CI of chromosome 17 in villous components of adenomas and all cancers was higher than that of normal epithelium (P < 0.05), but the differences were not significant. In protruding advanced cancers, the CI of chromosome 18 was significantly decreased (P < 0.01) compared to the CI of normal epithelium. There was no significant chromosomal heterogeneity between the superficial and the deepest layer in each cancer. In mucosa adjacent to sessile and flat type cancers, the CI of chromosome 17 was significantly higher than the CI in normal epithelium or adenomas (P < 0.05). These results suggest that numerical chromosome aberrations are associated with the histological type of adenoma and the morphological diversity of cancer in the colorectum, and that chromosome 17 abnormality occurs in mucosa adjacent to sessile and flat cancers.

Adenocarcinoma↗
PubMed2001

Quantitative analysis of 99Tcm-sestamibi myocardial perfusion SPECT using a three-dimensional reference heart: a comparison with experienced observers.

BACKGROUND: Quantification of myocardial perfusion single photon emission computed tomography (SPECT) may improve scintigraphic analysis. Recently, a fully operator independent technique for the quantification of myocardial perfusion SPECT was described, based on a normal three-dimensional averaged reference heart. The purpose of this study was to compare the automated SPECT quantification technique with experienced observers. METHODS: A total of 43 patients, 36 with one-vessel coronary artery disease (CAD) and seven with a low likelihood of CAD, underwent 99Tcm-sestamibi SPECT (99Tcm-MIBI SPECT). Three experienced observers and a panel (composed of the three observers), blinded to the clinical and angiographic data, analysed the size and severity of perfusion defects and the relation to the distribution areas of the coronary arteries. Inter-observer agreement was calculated by using kappa (kappa) statistics. RESULTS: The inter-observer agreement between the human observers and the automated quantitative analysis, for severity and size of perfusion abnormality, was moderate (kappa range 0.38-0.68), while this was fair between three individual observers (kappa range 0.36-0.87) and good between the individual observers and the panel (kappa range 0.63-0.89). There were no differences between the quantitative analysis and the panel in the allocation of perfusion abnormalities to the affected coronary artery. CONCLUSIONS: The operator independent quantification method showed a moderate agreement with individual observers and a panel analysis for size and severity of perfusion abnormalities. The automatic quantification has a similar ability to assign perfusion abnormalities to the diseased coronary artery as compared to an expert panel.

Adolescent↗
PubMed1984

Serial thallium-201 imaging after dipyridamole for coronary disease detection: quantitative analysis using myocardial clearance.

After dipyridamole, canine studies have demonstrated a slower rate of myocardial thallium-201 clearance from zones distal to a coronary artery stenosis compared to normal zones. To determine if criteria based on canine myocardial thallium-201 clearance rates could be applied clinically, 40 patients with and 26 patients without coronary artery disease (CAD) had serial thallium-201 images obtained for 2 to 5 hours after dipyridamole. Regions of interest were manually placed over six left ventricular segments in two projections for each of three imaging times. The myocardial thallium-201 clearance rate was calculated for each of the six segments and, using the clearance rate criterion found in canine studies, was considered abnormal if less than 6.5%/hr. Using this criterion alone, 22 of 26 patients (85%) without CAD had normal and 30 of 40 patients (75%) with CAD had abnormal myocardial thallium-201 clearance rates. A quantitative analysis of regional inhomogeneity in tracer distribution (normal was greater than or equal to 25% difference between segments) was negative in 24 of 26 patients (92%) without CAD and positive in 20 of 40 patients (50%) with CAD. When both clearance rate and regional inhomogeneity were considered, 21 of 26 patients (81%) without CAD had negative and 36 of 40 patients (90%) with CAD had positive results. Thus, post-dipyridamole myocardial clearance rate criteria derived from canine studies can be applied to clinical thallium imaging. Quantitative analysis of serial thallium-201 images after dipyridamole is optimized by using myocardial thallium-201 clearance rates. Such an approach is independent of regional inhomogeneities in tracer distribution.

Adult↗
PubMed1998

A simplified method for the quantitative analysis of 99Tc(m)-GSA liver scintigraphy using spectral analysis.

The aim of this study was to develop a simplified method for quantitative analysis of liver scintigraphy with 99Tc(m)-diethylenetriamine pentaacetic acid-galactosyl-human serum albumin (GSA) using spectral analysis. Dynamic liver scintigraphy using GSA was performed in three normal volunteers and 19 patients with chronic liver disease. Dynamic data were obtained with a gamma camera for 30 min after the injection of approximately 185 MBq GSA. The rate constant for the liver uptake of GSA from the blood (Ku, min(-1)), total excretion rate (Ke, min(-1)) and non-specific volume of distribution (Vh) were obtained by spectral analysis. Vh was defined as the volume in the liver region of interest (ROI) occupied by GSA which was in equilibrium with that in the blood. It should be noted that Vh had no units, since the counts in both the liver and heart ROIs were normalized by scan length to obtain counts pixel(-1) min(-). For comparison, compartmental analysis was also performed. A receptor index (LHL15) was calculated by dividing the radioactivity of the liver ROI by that of the liver plus heart ROIs 15 min post-injection. The Ku values obtained by spectral analysis (y) agreed well with those obtained by compartmental analysis (x) (y = 0.953x - 0.013, r = 0.992, S.E.E. = 0.016 min(-1)). The Ke and Vh values obtained by spectral analysis (y) correlated significantly with those obtained by compartmental analysis (x) (y = 1.149x - 0.016, r = 0.826, S.E.E. = 0.017 min(-1) for Ke; y = 1.191x + 0.044, r = 0.975, S.E.E. = 0.021 for Vh). The Ku values obtained by spectral analysis decreased as the severity of liver disease progressed, and were non-linearly related to the LHL15 values, suggesting that Ku is more sensitive to liver damage than LHL15, especially in the early stages of liver damage. These results suggest that spectral analysis applied to dynamic liver scintigraphy with GSA provides a simple, non-invasive and useful tool for the quantitative evaluation of liver function.

Hepatitis↗
PubMed1998

Quantitative analysis of sugar constituents of glycoproteins by capillary electrophoresis.

A method for quantitative analysis of monosaccharides including N-acetylneuraminic acid derived from sialic acid-containing oligosaccharides and glycoproteins is presented. The analysis is based on the combination of chemical and enzymatic methods coupled with capillary electrophoretic (CE) separation and laser-induced fluorescence (LIF) detection. The present method utilizes a simplified acid hydrolysis procedure consisting of mild hydrolysis (0.1 M TFA) to release sialic acid and strong acid hydrolysis (2.0 N TFA) to produce amino and neutral sugars. Amino sugars released from strong acid hydrolysis of oligosaccharides and glycoproteins were reacetylated and derivatized with 8-aminopyrene-1,3,6-trisulfonate (APTS) along with neutral sugars in the presence of sodium cyanoborohydride to yield quantitatively the highly stable fluorescent APTS adducts. N-acetylneuraminic acid (Neu5Ac), a major component of most mammalian glycoproteins, was converted in a fast specific reaction by the action of neuraminic acid aldolase (N-acylneuraminate pyruvate-lyase EC 4.1.3.3) to N-acetylmannosamine (ManNAc) and pyruvate. ManNAc was then derivatized with APTS in the same manner as the other monosaccharides. This method was demonstrated for the quantitation of pure Neu5Ac and the species derived from mild acid hydrolysis of 6'-sialyl-N-acetyllactosamine and bovine fetuin glycan. Quantitative recovery of the N-acetylmannosamine was obtained from a known amount of Neu5Ac in a mixture of seven other monosaccharides or from the sialylated oligosaccharides occurring in glycoproteins. The sequence of procedures consists of acid hydrolysis, enzymatic conversion and APTS derivatization which produced quantitative recovery of APTS-monosaccharide adducts. The detection limits for sugars derivatized with APTS and detected by CE-LIF are 100 pmol for Neu5Ac and 50 pmol for the other sugars.

Amino Sugars↗
PubMed2002

"Soap-Bubble" visualization and quantitative analysis of 3D coronary magnetic resonance angiograms.

In order to compare coronary magnetic resonance angiography (MRA) data obtained with different scanning methodologies, adequate visualization and presentation of the coronary MRA data need to be ensured. Furthermore, an objective quantitative comparison between images acquired with different scanning methods is desirable. To address this need, a software tool ("Soap-Bubble") that facilitates visualization and quantitative comparison of 3D volume targeted coronary MRA data was developed. In the present implementation, the user interactively specifies a curved subvolume (enclosed in the 3D coronary MRA data set) that closely encompasses the coronary arterial segments. With a 3D Delaunay triangulation and a parallel projection, this enables the simultaneous display of multiple coronary segments in one 2D representation. For objective quantitative analysis, frequently explored quantitative parameters such as signal-to-noise ratio (SNR); contrast-to-noise ratio (CNR); and vessel length, sharpness, and diameter can be assessed. The present tool supports visualization and objective, quantitative comparisons of coronary MRA data obtained with different scanning methods. The first results obtained in healthy adults and in patients with coronary artery disease are presented.

Adult↗
PubMed2005

Calibration-free quantitative analysis of mRNA.

Here we introduce a method for accurate and sensitive quantitative analysis of mRNA, which does not require calibration with mRNA. The method uses a fluorescently labeled hybridization probe as a reference standard. It involves the following: (i) annealing mRNA to the excess of the fluorescently labeled ssDNA hybridization probe, (ii) separation of the mRNA-probe hybrid from the excess of the probe by gel-free capillary electrophoresis mediated by ssDNA-binding protein, (iii) fluorescence detection of the hybrid and the excess probe, and (iv) quantification of mRNA using a simple algebraic formula. The method also overcomes a number of other limitations of conventional methods: the entire procedure currently takes only 2 h and accurately quantifies 10(5) copies of mRNA. With further improvements to the method, the procedure can be potentially shortened to 10 min, and the limit of quantification can be decreased to as few as 100 copies of mRNA. In this work, we prove the principle of the method by quantifying mRNA of green fluorescent protein in the matrix of total cellular RNA. The developed method is quantitative, simple, fast, and highly sensitive. It requires commercially available instrumentation only. The method will be an indispensable tool for molecular and cell biology studies.

Electrophoresis, Capillary↗
PubMed2001

Quantitative analysis of the elemental composition and the mass of bacterial polyphosphate bodies using STEM EDX.

The quantitative analysis of laboratory grown organisms (Plectonema boryanum and Staphylococcus aureus) revealed that a typical in vivo polyphosphate body (PPB) contains O (4.3 x 10(-8) microg), C (1.2 x 10(-8) microg), P (6.7 x 10(-9) microg), Mg (1.3 x 10(-9) microg), Ca (6.7 x 10(-10) microg), K (6.7 x 10(-10) microg), Fe (6.0 x 10(-10) microg), S (5.4 x 10(-10) microg) and Al (5.9 x 10(-10) microg). Quantitative X-ray analysis of samples from nature showed PPB contain O (1.63 x 10(-8) microg), C (4.75 x 10(-9) microg), P (2.50 x 10(-9) microg), Mg (5.0 x 10(-10) microg), Ca (2.50 x 10(-10) microg), K (2.50 x 10(-10) microg), Fe (2.25 x 10(-10) microg) and S (2.0 x 10(-10) microg). The mass of an average polyphosphate body was 6.7 x 10(-8) microg for P. boryanum, 2.5 x 10(-8) microg for S. aureus and for microbes from the natural environment 6.3 x 10(-8) microg. The results indicate that the PPB may have other unknown functions in addition to essential element storage, acting as a detoxification method by sequestering heavy metals and providing a homeostasis system in the cell.

Bacteria↗
PubMed1989

[Quantitative analysis of the lung CT in normal subjects and in patients with noncardiac pulmonary edema].

A quantitative analysis was performed of the CT images relative to 7 normal human subjects and to 22 patients affected with ARF (Acute Respiratory Failure). The CT scanner image quality was investigated, and the unit was checked with quality control procedures, in order to assure reliability and reproducibility. In every patient the lung was scanned at 3 different levels--basis, hilum and apex. The frequency distribution of CT numbers was studied. The lung areas were calculated for the different conditions, and the data correlated with functional lung data. A method was then tested to evaluate lung weight; the data obtained did not differ from literature data. All the patients were treated with different positive and expiratory pressure (PEEP). For different PEEP values, the quantitative data from CT analysis were correlated with morphofunctional indices.

Adult↗
PubMed1993

Repeated unilateral carotid occlusion in Mongolian gerbils: quantitative analysis of cortical neuronal loss.

To develop an experimental model which enables quantitative analysis of chronic neuronal loss in the cerebral cortex, repeated ischemic insult was performed using unilateral carotid artery occlusion in Mongolian gerbils. The effect of the time interval between the repeated ischemic insult on the survival rate of the animals and the amount of cortical neuronal loss were examined. The time course of the cortical neuronal damage in repeated ischemic insult was also studied. We repeated the occlusion four times; i.e., one 10-min and three 7-min occlusions (total 31 min of ischemia). The number of animals surviving for 3 weeks after the last ischemic insult was minimum (15.4%) for animals undergoing occlusions at 1-h intervals and maximum (100%) at 24- and 48-h intervals. The number of ischemic neuronal deaths was also dependent upon the time interval, and it was so pronounced as to allow analysis at intervals of 12 hr or 24 hr in the absence of infarction in the cortex. The number of neuronal deaths could not be determined for animals with occlusion at 1-h intervals due to the production of a large infarction, with which the 3-week survival rate was minimum. The temporal profile of cortical neuronal loss in the repeated ischemic insult at 24-h intervals indicated that the number of cortical neurons significantly decreased until 7 days after the start of the ischemic procedure. This model is useful for clarifying the pathophysiology of chronically developing ischemic neuronal death.

Animals↗
PubMed1998

Quantitative analysis of hyaluronan in the synovial tissues of patients with joint disorders.

Quantitative analysis of hyaluronan (hyaluronic acid; HA) in the synovial tissues of patients with joint disorders were performed. HA was found not only in the synovial intimal cells and matrices, but also especially in the alveolar lymphoid follicles and connective tissues surrounding blood vessels in the inflammatory granular synovium which formed the pannus. HA levels in the synovium of patients with rheumatoid arthritis (RA) (459.0+/-66.2 microg/g) where shown to be higher than those in patients with osteoarthritis (246.9+/-34.8 microg/g) and traumatic injury (227.7+/-35.4 microg/g). It follows from the present findings, HA in the synovium might contribute to the high amounts of serum levels of HA in patients with rheumatoid arthritis.

Adult↗
PubMed1986

Quantitative analysis of polyenoic phospholipid molecular species by high performance liquid chromatography.

The quantitative analysis of phospholipid molecular species containing polyenoic fatty acids is described. Dinitrobenzoyl derivatives of diacylglycerols prepared from phospholipids were separated into individual molecular species by reversed-phase high performance liquid chromatography (HPLC) using a combination of two solvent systems and were quantified at 254 nm. Thirty-six molecular species were resolved from the phosphatidylcholines of rat hearts, human platelets and Chinese hamster V79-R cells. The derivatives of alkenylacyl molecular species from platelet phosphatidylethanolamine were resolved concomitantly with diacyl molecular species.

Chromatography, High Pressure Liquid↗
PubMed1988

A method for the quantitative analysis of nerve growth in vitro.

A method is described for the quantitative analysis of the nerve-growth-promoting activity of biological molecules in tissue culture. The criteria used for the evaluation of this activity is based on the neurite length as well as the total number of neurites produced by the explant of whole dorsal root ganglia from 12-d-old chick embryos. A nerve growth index (NGI) is given to each ganglion during each of a 5-d culture period. The NGI is defined as the product of average neurite length in millimeters and the total number of neurites. We report that with increasing concentrations of fetal bovine serum, there was a proportional increase in NGI due to increased neurite density while the neurite length was not greatly affected. The NGI of several proteins with known nerve growth promoting activity, namely nerve growth factor, insulin, transferrin, and fibronectin were investigated for their activity and compared with that of fetal bovine serum.

Animals↗
PubMed1983

Quantitative analysis of the lateral geniculate nucleus in the mutant microphthalmic rat.

A quantitative analysis of the lateral geniculate nucleus was carried out in the mutant microphthalmic rat. In the dorsal lateral geniculate nucleus (LGNd) of the microphthalmic rat we found the total volume and neuronal population were reduced by 45 and 68% of normal values, respectively. The size of normal LGNd neurons was 8 to 20 microns and that of mutant LGNd cells from 6 to 16 microns. Neurons of the normal LGNd were medium-size and round or oval, and their cell bodies were filled with Nissl substance. Microphthalmic LGNd neurons, on the other hand, had narrow cytoplasmic spaces with few Nissl granules, and pale cell nuclei. In the microphthalmic rat, the lateral part of the ventral lateral geniculate nucleus (LGNvl) also showed a marked reduction in the total volume and neuronal population which were 42 and 76% of normal values, respectively. The size of normal LGNvl neurons was 8 to 20 microns and that of the microphthalmic neurons from 6 to 16 microns. These findings suggested that a marked reduction in the size of the LGNd and LGNvl in the mutant can be attributed to a decrease in neuronal population to a diminution of cell size.

Animals↗
PubMed1995

Quantitative analysis of synthetic human calcitonin by liquid chromatography-mass spectrometry.

The quantitative analysis of synthetic human calcitonin (hCT) by micro-liquid chromatography-electrospray ionization mass spectrometry (micro-LC-ESI-MS) is reported. hCT was extracted from plasma by an immobilized antibody column and separated by a micro-LC system. The molecular mass of hCT is 3417, and m/z 1140, corresponding to the [M + 3H]3+ ion, was observed by ESI-MS. This ion was monitored in the selected-ion monitoring mode; rat calcitonin, which is highly homologous to hCT, was used as an internal standard. The calibration curve for the quantification of hCT was linear in the range 10 ng/ml to 1 microgram/ml of plasma.

Amino Acid Sequence↗