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PubMed1999

Quantitative analysis of the dendritic architectures of cat hypoglossal motoneurons stained intracellularly with horseradish peroxidase.

Little is known about the dendritic architecture of cat hypoglossal motoneurons. Thus, the present study was done to provide quantitative descriptions of hypoglossal motoneurons and to determine correlations between dendritic size parameters by using the intracellular horseradish peroxidase (HRP) injection technique in the cat. Twelve hypoglossal motoneurons stained with HRP were antidromically activated by stimulation applied to the medial branch of hypoglossal nerve. Eight (type I) and four (type II) of the 12 motoneurons were located in the ventral and dorsal parts of the ventromedial subnucleus of hypoglossal nucleus, respectively. The somatodendritic morphology of the two types of neurons was remarkably different, especially in the dendritic arborization pattern. The type I neurons established an egg-shaped dendritic tree that was distributed through the nucleus to the reticular formation; the type II dendritic tree was confined within the nucleus and presented a rostrocaudally oriented, mirror-image, fan-shape appearance. The total dendritic area and length and the number of terminations and branch points were significantly larger for type I than for type II neurons. For the two types of neuron, there was a positive correlation between stem dendritic diameter and several dendritic size parameters. Although the slopes of the regression lines were slightly different between the two, these were not statistically significant. The present study provides evidence that hypoglossal motoneurons located in the ventromedial subnucleus could be divided into two types according to the dendritic arborization pattern and quantitative analysis of the dendritic tree and according to neuronal location and suggests that the two types of hypoglossal motoneurons can be viewed as intrinsically distinct cell types: type I and type II, which innervate extrinsic and intrinsic muscles, respectively. In addition, the morphometric analysis made it possible to estimate the size of the dendritic tree by measuring the stem dendritic diameter.

Animals↗
PubMed1989

[The stereologic-quantitative analysis of gastric antrum G cells in patients with splenic diseases].

In this paper, 89 Spleen deficiency (SD) patients' gastric mucosa specimens obtained during fibrogastroscopy mucosal biopsy were observed with immunocytochemical-stereologic-quantitative analysis method. The number of G cells counted and the content of basic gastrin in serum measured. This study found that the difference of the number of G cells and the content of gastrin in serum possessed statistical significance between two types of SD. The results suggested the hyperplasia and hypersecretion of G cells in gastric antrum might be an important pathological mechanism for different types of SD.

Adolescent↗
PubMed2006

Rapid method for quantitative analysis of the aroma impact compound, 2-acetyl-1-pyrroline, in fragrant rice using automated headspace gas chromatography.

A rapid method employing static headspace gas chromatography (HS-GC) has been developed and validated for quantitative analysis of the impact aroma compound, 2-acetyl-1-pyrroline (2AP), in grains of fragrant rice. This developed method excludes wet extraction, and the rice headspace volatiles are brought directly and automatically to GC analysis. The conditions of the static HS autosampler were optimized to achieve high recovery and sensitivity. The most effective amount of rice sample used was 1 g, which provided 51% recovery and a linear multiple headspace extraction (MHE) plot of the peak area of 2AP. The sensitivity of the method was enhanced by utilizing a megabore fused silica capillary column in conjunction with a nitrogen-phosphorus detector (NPD). Method validations performed for both static HS-GC-FID and HS-GC-NPD demonstrated linear calibration ranges of 20-10 000 (r(2) = 0.9997) and 5-8000 (r(2) = 0.9998) ng of 2AP/g of rice sample, respectively. The limits of detection for both systems were 20 and 5 ng of 2AP, and the limits of quantitation were 0.30 and 0.01 g of brown rice sample, respectively. Reproducibility calculated as intraday and interday coefficients of variation were 3.25% RSD (n = 15) and 3.92% RSD (n = 35), respectively, for SHS-GC-FID and 1.87% RSD (n = 15) and 2.85% RSD (n = 35), respectively, for SHS-GC-NPD. The method was found to be effective when applied to the evaluation of aroma quality, based on 2AP concentrations, of some fragrant rice samples.

Chromatography, Gas↗
PubMed2002

Quantitative analysis of chimerism after allogeneic peripheral blood stem cell transplantation.

BACKGROUND: For peripheral blood stem cell transplantation (PSCT), several engraftment analysis methods have been performed including detection of restriction fragment length polymorphisms and amplification of polymorphic genetic loci. To facilitate monitoring of the engraftment, a quantitative, non-isotopic method using a short tandem repeat (STR) marker has been set up in our laboratory. METHODS: DNAs from pretransplant recipients and donors were amplified with the AmpFISTR Profiler Plus kit that contains 9 STR markers. The fluorescent polymerase chain reaction products were then fractionated on polyacrylamide gels in an ABI PRISM 377 DNA sequencer. Results were analyzed using GeneScan 2.1 software. We selected the best markers as informative alleles which can distinguish donor from recipient. For quantitative analysis of the engraftment, we prepared a mixed chimeric sample by mixing pretransplant recipient and donor DNAs in different ratios to produce a standard curve. After amplifying the posttransplant recipient DNA, we were able to detect the extent of engraftment by interpolating the percent peak area of the informative alleles from this standard curve. RESULTS: We retrospectively analyzed 10 patients who had received allogeneic PSCT. Two of them showed some degree of mixed chimerism indicating leukemic relapse. In case one, 38.7% of the recipient DNA was first detected in the third month after PSCT. In case two, 6.5% of the recipient DNA was first detected in the tenth month after PSCT. CONCLUSION: In summary, this method provides an accurate, quantitative, and early assessment of mixed chimerism in posttransplant patients. Such information may be useful to guide implementation of additional treatment to circumvent graft failure or relapse in the future.

Adolescent↗
PubMed1984

A new model for the quantitative analysis of cell movements in vitro: definition of a shape change factor.

Changes in shape, in addition to translocations, are an important aspect of cell motility. We propose a simple geometrical model for the quantitative analysis of shape changes undergone by cultured cells. The extent to which images of a given cell do not overlap at the beginning and end of a time interval is used as a measure of motility, and a translation step included to eliminate translocation effects. Initial findings suggest that the method is widely applicable.

Animals↗
PubMed1982

Cytochemistry of adenylate cyclase. Quantitative analysis of the effect of cytochemical procedures on adenylate cyclase in heart tissue homogenates.

The effects of different preparative and cytochemical procedures on adenylate cyclase (AC) activity in heart muscle homogenates were studied by quantitative analysis. We were mainly concerned with perfusion prefixation, using glutaraldehyde (GA) with and without DMSO, and with the influence of cytochemical incubation with lead ions as the capture reagent. Furthermore, we measured the direct effect on the AC activity of lead, cobalt, and strontium ions in prefixed heart homogenates. We also studied the influence of phosphatidylinositol and 2',5'-dideoxyadenosine. The following results were obtained: 1. Perfusion fixation using 2% GA buffered with cacodylate reduced the AC activity by about 20%. After the entire cytochemical procedure was finished, 20% of the original AC activity was still present. Stimulation by epinephrine, histamine and fluoride, which increased the activity of AC two or three times in our experiments, was only slightly reduced by the cytochemical treatments. 2. Lead ions (2 mM), added to the biochemical assay without chelating compounds, reduced the AC activity about 90%. 5 muM phosphatidylinositol stabilized the fluoride- and hormone-sensitive AC activity. 3. Co2+ also reduced the AC activity, though less than Pb2+. Sr2+ had no effect on the basic activity of the AC but had a slightly inhibitory effect on the hormone and fluoride stimulation. 4. 5% DMSO added to the fixative had no influence on the basic activity of the AC. However, this solvent definitely reduced the level of stimulation by fluoride and guanine nucleotide plus hormones. 5. A potential inhibitor of enzyme activity and of the hormone- and fluoride-sensitive AC was the adenosine derivative, 2',5'-dideoxyadenosine. This compound, at a concentration of 10(-3) M, inhibited all AC activity in the heart homogenates.

Adenylyl Cyclases↗
PubMed1992

Cystatin activity in gingival crevicular fluid from periodontal disease patients measured by a new quantitative analysis method.

Cystatins are protein inhibitors of cysteine proteinases, which are believed to play an important role in the pathogenesis of periodontal disease. In this study, we report a new sensitive method for the quantitative analysis of cystatin activity in a small amount of crude sample such as gingival crevicular fluid. Cystatin activity in the crude sample was determined by using active site-titrated papain, which is a cysteine proteinase from the plant Carica papaya. Crude samples usually contain endogenous cysteine proteinases. These competed with the added papain for the active sites of the cystatins. The cystatin-cysteine proteinase complex was able to be dissociated by the addition of papain. This competition and dissociation could interfere with the determination of cystatin activity, since some of the cysteine proteinases, such as cathepsin B, hydrolyzed the specific substrate for papain during titration with the papain. In order to exclude this interference and measure total cystatin activity, the crude sample must be alkalinized (pH 11.0) for 5 min at 4 degrees C followed by 10 min at 40 degrees C before titration with papain. The minimum detectable amount of cystatins was 20 fmol/assay when it was calculated per mole of papain inhibitory sites. Using this method, significant levels of cystatin activity were detected in all the samples of gingival crevicular fluid taken from periodontal disease patients. These results suggest that cystatins could regulate the cysteine proteinases in gingival crevicular fluid and that this new method could be useful to clarify the role of cystatins in the pathogenesis of periodontal disease.

Alkalies↗
PubMed2001

Use of FT-NIR transmission spectroscopy for the quantitative analysis of an active ingredient in a translucent pharmaceutical topical gel formulation.

The objective of this study was to demonstrate the use of transmission Fourier transform near-infrared (FT-NIR) spectroscopy for quantitative analysis of an active ingredient in a translucent gel formulation. Gels were prepared using Carbopol 980 with 0%, 1%, 2%, 4%, 6%, and 8% ketoprofen and analyzed with an FT-NIR spectrophotometer operated in the transmission mode. The correlation coefficient of the calibration was 0.9996, and the root mean squared error of calibration was 0.0775%. The percent relative standard deviation for multiple measurements was 0.10%. The results prove that FT-NIR can be a good alternative to other, more time-consuming means of analysis for these types of formulations.

Administration, Topical↗
PubMed1992

[Spatial and quantitative analysis of the QRS and T waves by Frank-lead orthogonal electrocardiography in normal children. Comparative study in an early and late cord-clamping groups in newborns].

We studied a spatial and quantitative analysis of QRS and T waves by Frank-lead orthogonal electrocardiography (afterwards; ECG) in 50 cases of normal newborns, 30 cases of infants, 50 cases of adults, and also the variation in age was investigated. In the newborns, the comparative study was made serially, longitudinally in 21 infants with an early clamping of the cord and 29 infants with a late clamping of the cord. The results are mainly as follows. 1) The parameters reflecting the cardiac electromotive force of the right ventricle such as Sx, Qz and Sx+Qz decreased with aging. 2) Rx, i.e., the maximum leftward projection of QRS vector increased until the children, probably because the left ventricle was dominant in their growth and development and in the increased amount of work. A subsequent reduced value in the adults was considered to be mainly due to an increase of the distance between the heart and electrode by the increase in the thorax, particularly the transverse diameter and due to the growth in subcutaneous fat and muscle. 3) The values of Rx/Sx increased with aging. Rx and Sx in the adults were less than those in the children, but Rx/Sx was larger than that of Rx because Sx was less than Rx. 4) Age-related changes of Tz was most characteristic and was negative up to several days after birth. T vector was in forward direction and then turned positive (in backward). This was greatest at the period of the infants. Subsequently, it turned negative in the childhood, and extent was maximum in the adults.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed2004

Computer-assisted quantitative analysis of bone marrow edema of the knee: initial experience with a new method.

OBJECTIVE: The purpose of this study was to describe a largely observer-independent computer-assisted method for accurate quantitative analysis of bone marrow edema. MATERIALS AND METHODS: Ten patients with bone marrow edema of the knee were included in the study. Coronal STIR images of the affected knees were obtained using a 1.0-T MR scanner. Size and signal intensity of the bone marrow edema were assessed on the basis of gray-scale value analysis and calculation of a threshold value for differentiating normal and edematous bone marrow. All measurements were carried out three times for statistical analysis. RESULTS: The intraobserver coefficient of variation was 0.89% for the volume and 0.94% for the signal intensity of the bone marrow edema, showing the small impact of manual interference on results produced with this method. CONCLUSION: A computer-assisted method for quantification of bone marrow edema has been described. Intraobserver variation was very low, indicating excellent reproducibility of results. Although the method is too time-consuming for clinical use, it is recommended for research purposes.

Bone Marrow Diseases↗
PubMed1979

[Quantitative analysis of L-hydroxyproline in human biological material by gas-chromatography (author's transl)].

Quantitative amino acid analysis by gas chromatography was applied to the determination of 4-hydroxy-L-proline in biological material. After addition of a known amount of allo-4-hydroxy-D-proline, quantitative analysis of 4-hydroxy-L-proline can be performed by gas chromatographic separation of the amino acid enantiomers; since the standard and sample have identical chemical behaviour, they respond identically during derivatization. The accuracy, reproducibility, and sample requirement of this procedure are more advantageous than those of hitherto existing methods. This study is also an example of the application of chiral stationary phases, which permit the simultaneous separation of amino acids and amino acid enantiomers.

Chromatography, Gas↗
PubMed2006

A parallel-gradient microfluidic chamber for quantitative analysis of breast cancer cell chemotaxis.

Growth factor-induced chemotaxis of cancer cells is believed to play a critical role in metastasis, directing the spread of cancer from the primary tumor to secondary sites in the body. Understanding the mechanistic and quantitative behavior of cancer cell migration in growth factor gradients would greatly help in future treatment of metastatic cancers. Using a novel microfluidic chemotaxis chamber capable of simultaneously generating multiple growth factor gradients, we examined the migration of the human metastatic breast cancer cell line MDA-MB-231 in various conditions. First, we quantified and compared the migration in two gradients of epidermal growth factor (EGF) spanning different concentrations: 0-50 ng/ml and 0.1-6 ng/ml. Cells showed a stronger response in the 0-50 ng/ml gradient. However, the fact that even a shallow gradient of EGF can induce chemotaxis, and that EGF can direct migration over a large dynamic range of gradients, confirms the potency of EGF as a chemoattractant. Second, we investigated the effect of antibody against the EGF receptor (EGFR) on MDA-MB-231 chemotaxis. Quantitative analysis indicated that anti-EGFR antibody impaired both motility and directional orientation (CI = 0.03, speed = 0.71 microm/min), indicating that cell motility was induced by the activation of EGFR. The ability to compare, in terms of quantitative parameters, the effects of different pharmaceutical inhibitors, as well as subtle differences in experimental conditions, will aid in our understanding of mechanisms that drive metastasis. The microfluidic chamber described in this work will provide a platform for cell-based assays that can be used to compare the effectiveness of different pharmaceutical compounds targeting cell migration and metastasis.

Breast Neoplasms↗
PubMed1990

Quantitative analysis of 18/FDG-PET in the presurgical evaluation of patients suffering from refractory partial epilepsy. Comparison with CT, MRI, and combined subdural and depth. EEG.

CT, MRI, 18/FDG-PET and Depth. EEG, performed with subdural and depth electrodes were part of the presurgical evaluation in 22 patients. Statistical analysis of 18/FDG-PET was performed to compare cerebral utilization of glucose to that of normal age matched controls. The findings of CT, MRI, and quantitative analysis of PET are compared with those of ictal Depth. EEG. A positive correlation between CT and Depth. EEG was obtained in 23% of the patients and between MRI and Depth. EEG. in 50%. For both imaging techniques a negative correlation was found in 5%. Regional abnormalities were found with quantified PET in 95% of the patients and were concordant with Depth. EEG. for side of onset in 77% of the patients and for lobe of onset in 59%. A possibly false localising PET result for lobe of onset was obtained in 8 patients (36%). Limitations of PET were most apparent in patients with regional mesiolimbic or bilateral seizure onset. A favourable outcome of surgery was associated usually with positive convergence of both methods. PET may be a valuable contribution to the research and management of partial complex epilepsy, but at present cannot be considered a reliable alternative to invasive EEG methods in patients without clear unilateral focus localization on surface EEG.

Adolescent↗
PubMed1986

Quantitative analysis of quadriceps muscle biopsy. Results in 7 definite and 45 possible carriers of Duchenne muscular dystrophy.

Quadriceps muscle biopsy was performed in 7 definite and 45 possible carriers of Duchenne muscular dystrophy. Unequivocal morphological changes were observed in 8 women; all but one of them were already detected as carriers by increased serum CK. Histometric analysis of muscle biopsy was performed on two fibre types (type 1 and type 2) in all 52 women; in 38 of them the two subgroups of type 2 fibres (type 2a and type 2b) were also quantified. Results of histometric analysis were matched with control values obtained from 30 healthy females in the same age as the carriers (20-50 yr). Histometric analysis on two fibre types detected a percentage of carriers (27%) much lower than that detected by serum CK determination (44%). The detection rate of combined serum CK determination and histometric analysis of muscle biopsy was: 100% in definite carriers, 65% in mothers possible carriers, 38% in sisters and 25% in the group of cousins and aunts. The histometric parameters most frequently altered were atrophy factor and variability coefficient of mean diameter. Histometric analysis carried out on three fibre types in 38 women detected 2 more carriers showing atrophy of type 2b fibres. The results indicate that quantitative analysis of muscle biopsy may be used to detect some possible carriers with normal serum CK. However, an adequate number of controls is mandatory; in fact the detection rate of histometric analysis of muscle biopsy decreased by increasing the number of controls, resulting much lower than previously reported.

Adult↗
PubMed1991

Quantitative analysis of the molecular species of monosialogangliosides by continuous-flow fast-atom bombardment mass spectrometry.

Negative-ion continuous-flow fast-atom bombardment mass spectrometry was evaluated as a means for the quantitative analysis of N-acetylneuraminyl-galactosyl-glucosyl-ceramide (NeuAc-GM3) and N-acetylgalactosaminyl-(N-acetylneuraminyl)galactosyl-glucosyl-ceramide (NeuAc-GM2). This study was carried out on a 7070-EQ mass spectrometer (VG Analytical, Manchester, UK) using a home-made continuous-flow fast-atom bombardment probe with a mixture of methanol + water + triethanolamine (70:27:3, v/v/v) as the mobile phase. Utilizing 100 ng of acetyl-lysogalactosyl-N-acetylgalactosaminyl-(N-acetylneuraminyl)g alactosyl-glucosyl-ceramide (acetyl-lysoGM1) as an internal standard, standard curves for NeuAc-GM3 d18:1-16:0, NeuAc-GM3 d18:1-18:0 and Neuac-GM2 d18:1-18:0 were found to be linear over the range 5-250 ng, with associated correlation coefficients of 0.990-0.997. The lower limit of detection was found to be 2.5 ng. Satisfactory results could also be obtained when the calibration curves were derived from the deprotonated molecular ions of a mixture of the NeuAc-GM2 and NeuAc-GM3 classes. Using this approach, quantitative determination of NeuAc-GM3 d18:1-16:0 from rat adrenal gland was performed using N-acetylneuraminic acid assay as a test control. We found 278 +/- 36 ng of this species in 1 mg of tissue (three replicate experiments). The procedure represents a sensitive method for the quantitation of monosialogangliosides and its capability to give molecular species information.

Adrenal Glands↗
PubMed1985

Three-dimensional reconstruction and quantitative analysis of rat lung type II cells: a computer-based study.

The three-dimensional structure of alveolar epithelial type II cells was imaged using a computer-based system designed for reconstruction and quantitative analysis of serially sectioned specimens. Six type II cells were reconstructed from serial ultrathin sections of lungs from two Sprague Dawley male rats and the results were compared to standard morphometric estimates of type II cell composition from five other Sprague Dawley male rats. A minor portion of the type II cell surface was in contact with the alveolar airspace while most of the cell surface was embedded in the alveolar septal interstitium. The type II cells contained multiple Golgi regions located close to the nucleus. Mitochondria formed a few branching filamentous networks extending throughout the cell. The reconstructed cells appeared to represent a homogeneous population having fractional volumes of intracellular organelles very similar to those found by morphometric techniques. The spatial distribution of secretory organelle volume suggests that the organization of this cell type reflects an ordered progression of secretory particle maturation which is consistent with earlier hypotheses of lamellar body assembly.

Animals↗
PubMed2002

Quantitative analysis using gas chromatography/combustion/isotope ratio mass spectrometry and standard addition of intrinsically labelled standards (SAIL)--application to isoflavones in foods.

We have investigated a novel application of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) for the quantitative analysis of the isoflavones in food matrices. Previous methods have been hampered by the lack of analytical standards to introduce early enough in the extraction procedure to allow compensations for losses at all stages of the procedure. In this work we have produced standard materials that can be added at the initial extraction, by intrinsically labelling soya plants by growing them in an atmosphere enriched in the stable isotope of carbon in CO(2). On analysis these plants were shown to contain phytoestrogens at a high (up to 20%) level of enrichment. The dried plant material has been used to estimate the isoflavone concentrations of a set of spiked flours. For daidzein the methodology was shown to produce results comparable to those achieved by GC/MS techniques. The method was less successful for genistein, possibly due to the greater fragility of this compound under the conditions required for the analysis.

Carbon Isotopes↗
PubMed2003

[Quantitative analysis of urban forest structure: a case study on Shenyang arboretum].

The spatial features and attribute data of urban forests in Shenyang arboretum were acquired by using aerial photographs and field investigation. After systemic and quantitative analysis of the species composition and spatial distribution pattern of urban forest community based on geographic information system (GIS), the following results were achieved: 1) the vegetation in the arboretum was rich and in a good condition. Phellodendron amurense, Robinia pseudoacacia, and Ulmus punita were the main species according to their abundant individuals and relative dominance. The spatial distribution pattern of this vegetation indicated that it was in a growing level and in the best state for ecological benefits; 2) the tree species relative dominance based on vegetation quantity had no distinct relationship with the species abundance, and the species having abundant individuals might have a low relative dominance; and 3) the curve displayed in the relationship diagram between tree individuals and tree height showed ascending and protruding. With the increment of tree height class, the individuals were unchanged nearly at the beginning, but when the height was more than 12 m, the individuals reduced distinctly, and the curve displayed a quickly declined trend. The relationship between tree individuals and tree diameter at the breast height (DBH) showed the same result.

City Planning↗