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PubMed2002

Quantitative analysis of two-component samples using in-line hard X-ray images.

Methods for rapid quantitative phase-sensitive X-ray imaging of non-crystalline samples consisting of two distinct components are investigated. The transverse spatial distribution of the projected thickness of each component is reconstructed by computer processing of in-line images collected using synchrotron-generated hard X-rays and a position-sensitive detector with submicrometre spatial resolution. Different imaging techniques and associated image-processing algorithms are considered, with relative advantages and difficulties of each approach compared. A possible generalization of the method for the case of n-component samples is briefly discussed.

Journal Article↗
PubMed1983

Quantitative analysis of eumelanin and pheomelanin in hair and melanomas.

In this study, a method is provided for analyzing quantitatively the content and the class of melanin pigments in the tissues, e.g., hair and melanoma. The method is simple and rapid because it does not require the isolation of melanins from the tissues. The rationale was that permanganate oxidation of eumelanin yields pyrrole-2,3,5-tricarboxylic acid (PTCA) as its major pyrrolic product, which may serve as a quantitatively significant indicator of eumelanin, while hydriodic acid hydrolysis of pheomelanin yields amino-hydroxyphenylalanine (AHP) as a specific indicator of pheomelanin. The degradation products, PTCA and AHP, were determined by high-performance liquid chromatography. Sepia melanosome-melanin and synthetic 5-S-cysteinyldopa-melanin served as reference standards of eumelanin and pheomelanin, respectively. Our method provided data that corresponded well to the content and class of melanins in normal hair. Based on this control study, it was found that the melanins in the melanosomes of both B16 and Harding-Passey (HP) melanomas were eumelanic and that the melanin content in B16 melanosomes was more than 10 times higher than that in HP melanosomes, though these two melanosomes revealed distinct colors and ultrastructures, i.e., brown-black, eumelanosome-like granules in B16 and reddish- or light-brown, pheomelanosome-like granules in HP.

Animals↗
PubMed2000

Quantitative analysis of human herpesvirus 8 viral load using a real-time PCR assay.

We have developed a quantitative real-time PCR (TaqMan) assay aimed at measuring the cellular human herpesvirus 8 (HHV-8) DNA load in various clinical samples. Standard curves were obtained by serial dilutions of a control plasmid containing both HHV-8 (ORF73 gene) and the cellular target (human albumin gene). The assay appeared to be very sensitive (100% detection rate for at least 10 copies per well) and specific and was easily reproducible (less than 3% intra-assay variability, 5% interassay variability). This method allowed us to quantify precisely the average HHV-8 copy number per cell in various persistently HHV-8-infected cell lines (BBG-1 cells, n = 200; BC-1 cells, n = 59; BCBL-1 cells, n = 70). A retrospective study was also conducted to assess the HHV-8 DNA load in 12 human immunodeficiency virus-infected patients with either Kaposi's sarcoma (KS; seven patients monitored over a 3-month period) or multicentric Castleman's disease (MCD; five patients). The HHV-8 DNA load ranged from 0 to 9,171 copies/10(6) cells in low-risk KS patients (T0, I0, S0 according to the classification of the AIDS Clinical Trials group). We also measured the viral loads in MCD patients either during symptomatic periods or during remission. The results are in agreement with previously published data, with high viral loads correlating with clinical symptoms (1.3 x 10(6) copies/10(6) cells) and low viral loads correlating with asymptomatic periods (less than 5,000 copies/10(6) cells).

AIDS-Related Opportunistic Infections↗
PubMed1976

Sphincterotomy and micturition parameters: quantitative analysis of sphincterotomy in 24 patients.

The effect of transurethral external sphincterotomy, performed in 24 patients, was evaluated quantitatively on micturition parameters and blood loss. The total urethral resistance, residual urine, and the maximum and average urinary flow rates were all improved with a statistically significant difference (p less than 0.01). Of 7 patients with indwelling catheter, all but one became catheter free. Erythrocytes and hemoglobin decreased 6% in number and 9% in weight, respectively. Moderate degree of stress incontinence were noted in 15 patients (63%).

Adult↗
PubMed1981

Evaluation and prediction of regenerative processes by quantitative analysis of the electromyographic response and biofeedback in peripheral nerve injuries.

The purpose of this study was to quantitatively and objectively evaluate and monitor regenerative processes in peripheral nerve lesions using a new electrophysiological method, and to develop and evaluate an electromyographic (EMG) biofeedback training programme for the rehabilitation of patients with such injuries. Five patients with traumatic unilateral brachial plexus injuries participated in the study longitudinally on a twice-weekly basis. The electrophysiological method was based on quantitative on-line analyses of the myoelectric signal from affected muscles of the upper limb. The biofeedback programme was developed and used to: (1) increase maximum voluntary centrally mediated neuromuscular activity; (2) dissociate desired and interfering neuromuscular activity; and (3) improve patient control over the neuromuscular activity. The electrophysiological results have differentiated clearly between muscles where regeneration can be assumed to be in progress and muscles where it is not in progress. This differentiation can be made prior to the occurrence of any observable contraction in muscle under study. The biofeedback training resulted in improved neuromuscular function in all patients.

Brachial Plexus↗
PubMed1992

[Use of Hoechst fluorescent probe 33258 for quantitative analysis of intracellular binding of anthracycline antibiotics to DNA].

The paper deals with development of a procedure for quantitative determination of the share of anthracycline antibiotics bound in cells directly to DNA. A DNA-specific Hoechst fluorescence dye 33258 was used for the purpose. The level of its quenching on DNA correlated with the quantity of the antibiotic bound to it. It was shown that the quenching of the Hoechst fluorescence dye bound to DNA was not due to the dye competition with the antibiotic for the site of bounding on DNA, as was suggested earlier. It was likely to be defined by reabsorption of the radiation by antibiotic molecules.

Animals↗
PubMed1991

[A method of quantitative analysis of bendiocarb in the air of the work area].

Proposed was a highly sensitive technique for a quantitative assessment of the newly developed insecticide bendiocarb++ in the working zone air (0.1 hg in the given air sample, or 0.025 mg/m3). The technique was tested in toxicological studies of bendiocarb++ contents in the working zone air.

Air Pollutants, Occupational↗
PubMed1991

Slipped capital femoral epiphysis. A quantitative analysis of motion, gait, and femoral remodeling after in situ fixation.

A prospective clinical study was done for quantitative examination of motion of the hip, gait, and proximal femoral remodeling after in situ fixation of a slipped capital femoral epiphysis. In situ fixation was performed in forty-five consecutively treated patients (fifty-six hips). Thirty-nine (87 per cent) of the patients returned for examination two years after treatment. The greatest percentage of motion of the hip returned within six months after treatment. Despite loss of internal rotation of the hip, the mean foot-progression angle was 10.8 degrees. Radiography and computerized tomographic scanning revealed minimum change in the relationship of the femoral head to the femoral shaft and no change in the neck-shaft angle. Motion returned despite minimum osseous remodeling. The early return of motion (in the first three months) may have been due to relief of pain, spasm, and synovitis, while soft-tissue stretching and resorption of bone in the anterolateral part of the femoral neck may have accounted for the remainder of the increase in internal rotation.

Adolescent↗
PubMed1986

Quantitative analysis of phospholipids by 31P-NMR.

High-field 31P nuclear magnetic resonance spectroscopy was used to quantitate phospholipids in mixtures in organic solvents. The sample is dissolved in chloroform-methanol and analyzed at 161.7 MHz with decoupling of the protons. Signals were identified using authentic compounds, and their relative distribution was measured in mole percent. The method has good accuracy and reproducibility, and was used to analyze phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, lysophosphatidylcholine, lysophosphatidylethanolamine, phosphatidylinositol, cardiolipin, and phosphatidic acid in egg lecithin. Four commercial egg phospholipids and the phospholipids from a total lipid extract of rat liver were analyzed. The method could be utilized to analyze phospholipids from other sources.

Animals↗
PubMed1994

[Quantitative analysis for histochemical grouping of muscle fibers after skeletal muscle transplantation by microneurovascular anastomoses].

A poly-histochemical quantitative assay was carried out on fibers of grafted rectus femoris muscles of rabbit which had undergone simulated free muscle transplantation by microneurovascular anastomoses in speciments at 3, 6, 12 months postoperatively. At the same time, the same assay was done for two experimental control groups: either the motor nerve or the patella tendon was simply severed and immediately sutured. It was found that the contractile characteristics of whole muscle were depended on relative number of different muscle fibers. That meant it was depended on the relative number of different motor fibers which had run through the anastomosed site. The caliber change among different types of muscle fibers mainly represented there was cross reneurotization during nerve regeneration.

Anastomosis, Surgical↗
PubMed1993

Quantitative analysis of alternative splicing options of human plasma membrane calcium pump genes.

The alternative splicing options and the quantitative tissue distribution of the transcripts of the four currently known human plasma membrane calcium pump (PMCA) genes have been analyzed in seven tissues (cerebral cortex, skeletal and heart muscle, stomach, liver, lung, and kidney) by quantitative polymerase chain reaction on reverse transcribed mRNA with glyceraldehyde-3-phosphate dehydrogenase as the internal standard. The mRNAs of genes 1 and 4 were found to be present in similar amounts in all tissues, whereas the transcripts of genes 2 and 3 were expressed in a tissue-specific manner, i.e. their amounts were highest in fetal skeletal muscle and brain. Alternative splicing was found to occur in the PMCA transcripts at two major regulatory sites (sites A and C), adjacent to the amino-terminal phospholipid-responsive region and within the carboxyl-terminal calmodulin binding domain, respectively. Novel splicing variants not described previously for human genes were detected for hPMCA3 and 4 at site A and for hPMCA1, 2, and 3 at site C. For all genes a common splice variant was found at both splice sites. The common splice variant at site A was characterized by the inclusion of a small exon (hPMCA1, 39 base pairs (bp); hPMCA2, 42 bp; hPMCA3, 42 bp; hPMCA4, 36 bp). In the common splice variant at site C, an exon (hPMCA1, 154 bp; hPMCA2, 227 bp; hPMCA3, 154 bp; hPMCA4, 178 bp) was excluded in the mRNA. All genes normally express these main splice variants in all tissues in which the corresponding isoform is present. The splicing complexity at site C was found to be augmented in the transcripts of PMCA2 and PMCA3 through the use of additional exons, and in PMCA1 and 3 through the use of additional internal splice sites in the single alternatively spliced 154-base pair exon.

Aged↗
PubMed2002

Starch biosynthesis and intermediary metabolism in maize kernels. Quantitative analysis of metabolite flux by nuclear magnetic resonance.

The seeds of cereals represent an important sink for metabolites during the accumulation of storage products, and seeds are an essential component of human and animal nutrition. Understanding the metabolic interconversions (networks) underpinning storage product formation could provide the foundation for effective metabolic engineering of these primary nutritional sources. In this paper, we describe the use of retrobiosynthetic nuclear magnetic resonance analysis to establish the metabolic history of the glucose (Glc) units of starch in maize (Zea mays) kernels. Maize kernel cultures were grown with [U-(13)C(6)]Glc, [U-(13)C(12)]sucrose, or [1,2-(13)C(2)]acetate as supplements. After 19 d, starch was hydrolyzed, and the isotopomer composition of the resulting Glc was determined by quantitative nuclear magnetic resonance analysis. [1,2-(13)C(2)]Acetate was not incorporated into starch. [U-(13)C(6)]Glc or [U-(13)C(12)]sucrose gave similar labeling patterns of polysaccharide Glc units, which were dominated by [1,2,3-(13)C(3)]- and [4,5,6-(13)C(3)]-isotopomers, whereas the [U-(13)C(6)]-, [3,4,5,6-(13)C(4)]-, [1,2-(13)C(2)]-, [5,6-(13)C(2)], [3-(13)C(1)], and [4-(13)C(1)]-isotopomers were present at lower levels. These isotopomer compositions indicate that there is extensive recycling of Glc before its incorporation into starch, via the enzymes of glycolytic, glucogenic, and pentose phosphate pathways. The relatively high abundance of the [5,6-(13)C(2)]-isotopomer can be explained by the joint operation of glycolysis/glucogenesis and the pentose phosphate pathway.

Carbon Isotopes↗
PubMed1982

Complexes of cytochrome P450 with metyrapone. A convenient method for the quantitative analysis of phenobarbital-inducible cytochrome P450 in rat liver microsomes.

Phenobarbital-inducible cytochrome P450 variants have a higher affinity (app. Ks = 0.6 microM). The difference in affinity has enabled us to quantitate, by metyrapone-induced difference spectral analysis, phenobarbital-inducible cytochromes P450 in liver microsomal membranes. The result obtained by this simple, rapid technique compare well with those found by more sophisticated techniques such as radioimmunoassay and immunoprecipitation.

Animals↗
PubMed1998

Quantitative analysis of HBY 097 and its metabolites in human serum and urine by HPLC.

Two HPLC methods were developed: one for the quantitation of HBY 097 reverse transcriptase inhibitor and its metabolites M2 and M3 in human serum, and one for the quantitation of metabolite M5 in urine. The HPLC procedure for the quantitation of HBY 097 and its metabolites M2 and M3 in human serum involved protein precipitation with acetonitrile followed by automated on-line trace enrichment. The HPLC procedure for the analysis of metabolite M5 in urine involved enzymatic hydrolysis of urine with beta-glucuronidase to convert metabolite M5 (glucuronide of M3) to M3. Reverse phase chromatographic separation with gradient elution. UV detection at 335 nm, and internal standard were used to quantitate analytes in both procedures. The lower quantitation limits were 25 ng ml-1 for HBY 097 and metabolites M2 and M3 in serum, and 0.5 microgram ml-1 for the metabolite M5 in urine measured as metabolite M3 after hydrolysis. The HBY 097 and metabolite M3 concentrations were specific but metabolite M2 was semi-specific because the two diastereomers of M2 were not resolved by the present chromatographic procedure. Both procedures were applied to the quantitation of HBY 097 and its metabolites in serum and urine of HIV positive patients who were enrolled in a clinical study of drug safety and pharmacokinetics.

Antiviral Agents↗
PubMed2005

Quantitative analysis of genetic and neuronal multi-perturbation experiments.

Perturbation studies, in which functional performance is measured after deletion, mutation, or lesion of elements of a biological system, have been traditionally employed in many fields in biology. The vast majority of these studies have been qualitative and have employed single perturbations, often resulting in little phenotypic effect. Recently, newly emerging experimental techniques have allowed researchers to carry out concomitant multi-perturbations and to uncover the causal functional contributions of system elements. This study presents a rigorous and quantitative multi-perturbation analysis of gene knockout and neuronal ablation experiments. In both cases, a quantification of the elements' contributions, and new insights and predictions, are provided. Multi-perturbation analysis has a potentially wide range of applications and is gradually becoming an essential tool in biology.

Algorithms↗
PubMed1998

The quantitative analysis of crystallinity using FT-Raman spectroscopy.

PURPOSE: To establish if FT-Raman spectroscopy can be used to quantitate the degree of crystallinity in a model compound. METHODS: Mixtures containing different proportions of amorphous and crystalline indomethacin were prepared. Using the peak intensity ratio 1698 cm(-1) (crystalline) to 1680 cm(-1) (amorphous), a correlation curve was prepared. This correlation curve was validated by testing further samples of known composition. Partially crystalline indomethacin was prepared by milling crystalline indomethacin. RESULTS: A linear correlation curve was obtained across the entire range of 0-100% crystallinity. Using this method, it was possible to detect down to either 1% amorphous or crystalline content. The largest errors were found to result from inhomogeneities in the mixing of the calibration and validation samples. The spectra of the mechanically processed samples were similar to the spectra of the calibration samples, and the degree of crystallinity could be estimated in these samples. CONCLUSIONS: FT-Raman spectroscopy is a potentially useful method to complement existing techniques for the quantitative determination of crystallinity.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2004

[Spatial variability and quantitative analysis of field factors based on GIS].

The objective of this research was to investigate the variability and the quantitative relationships among soil nutrients and crop growth status and yield. All data were analyzed by both classical statistics and geostatistics based on GIS. Soil properties included soil pH, total N, organic matter, available P and available K, while crop growth status was indicated by SPAD, LAI and SPAD x LAI. All parameters except soil pH exhibited spatial correlation. Soil total N and organic matter, SPAD, LAI and SPAD x LAI were all correlated to rice yield. Kriged interpolation maps provided good indication of the spatial variability in crop yield and growth status. Spatial interpolation and correlation analysis proved that SPAD x LAI was more indicative of crop growth status than individual variables, and useful for implementing growth season and topdressing as needed.

Biomass↗
PubMed1982

Inhibition of axoplasmic transport in the developing visual system of the rat-II, Quantitative analysis of alterations in transport of tritiated proline or fucose.

Developmental alterations in the amount of tritiated proline and fucose incorporated by retinal neurons and transported within axons of the optic nerve to the dorsal lateral geniculate nucleus and superior colliculus were measured at 1, 5, 10 and 15 days postnatal using quantitative autoradiography and liquid scintillation analysis. The amount of axon transport inhibition induced by introacular injections of colchicine (10-4 M-5 X 10-3M) and xylocaine (10-3 M-10-1 M) was also determined by this methodology. Grain counts of retinal autoradiograms obtained from animals of all ages employed in this study demonstrated that [3H] proline is rapidly incorporated by all retinal neurons but becomes increasingly concentrated within the inner nuclear, ganglion cell and optic fiber layers between 2h and 2 days after injection. [3H] fucose is preferentially taken up and concentrated within the plexiform and sensory element layers. Intraocular colchicine administered 24 h prior to isotope injection exhibited no significant effect on incorporation but depressed the amount of activity in the layer of optic fibers. Comparison of the effects of colchicine and xylocaine on axon transport of [3H] proline injected at 3 days of age revealed dose-dependent suppression of transport occurring up to six hr after isotope injection; however, with longer survival periods the effects of xylocaine were no longer significant whereas colchicine maintained suppression of axon transport to 20% of normal for periods of up to 10 days. Additionally, the rate and quantity of 3H] proline transported to the dorsal lateral geniculate nucleus and tectum of 1-15 day old animals was found to be inversely proportional to the age of the animal. Maximally-effective concentrations of colchicine, determined for each age level examined by the previous study to be highest compatible with the viability of the retino-fugal projection, 34 also reduced transport to from 20-40% of normal, being most effective in animals of 1-10 days of age. Autoradiographs of the optic disc demonstrated a colchicine-induced suppression of the amount of label in the exiting optic fibers and similar preparations of the tectum revealed an elimination of activity in the stratum griseum superficialis and stratum opticum which are normally heavily labeled following intraocular [3H] proline or fucose. Following [3H] fucose injection at 10 days of age, isotope appears in the contralateral visual cortex between 3 and 10 days later, indicating a substantial amount of transneuronal transfer of label which appeared to occur in concert with periods of maximal synaptogenesis. Injection of colchicine reduced the amount of label in the cortex to background levels. Concomitantly, autoradiographs of the dorsal lateral geniculate nucleus revealed cytoplasmic labeling of geniculate neurons in control animals but this was eliminated following colchicine injection...

Animals↗