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At least 703 records · Page 39Linked to original sources
PubMed2006

[Quantitative analysis of epidermal growth factor receptor mRNA in laryngeal carcinoma by reverse transcription-real time polymerase chain reaction with TagMan probe].

OBJECTIVE: To establish a method of quantitative analysis of epidermal growth factor receptor(EGFR) mRNA in laryngeal carcinoma by reverse transcription-real time polymerase chain reaction with TagMan probe and evaluate its applicable value. METHODS: The technique of reverse transcription-real time polymerase chain reaction with highly specific primers and probe was applied. And with GAPDH as internal standard, EGFR mRNA in 32 laryngeal carcinoma tissues, compared with their own macroscopically normal laryngeal mucosa tissues adjacent to the tumors, were quantitatively examined. The recombinant plasmid of EGFR was constructed, and then sequenced. RESULTS: The recombinant plasmid of EGFR was successfully constructed. And the mRNA index of EGFR in 32 laryngeal carcinoma tissues in terms of M (QR) was 0.025 (0.076), which was higher than 0.008 (0.027) in their own macroscopically normal laryngeal mucosa tissues adjacent to the tumors, P value is 0. 007. CONCLUSIONS: The method of reverse transcription-real time polymerase chain reaction with TagMan probe is quite objective, precise and high throughput in the application for the mensuration of EGFR mRNA in laryngeal carcinoma.

Adult↗
PubMed2006

Augmented vessels for quantitative analysis of vascular abnormalities and endovascular treatment planning.

Endovascular treatment plays an important role in the minimally invasive treatment of patients with vascular diseases, a major cause of morbidity and mortality worldwide. Given a segmentation of an angiography, quantitative analysis of abnormal structures can aid radiologists in choosing appropriate treatments and apparatuses. However, effective quantitation is only attainable if the abnormalities are identified from the vasculature. To achieve this, a novel method is developed, which works on the simpler shape of normal vessels to identify different vascular abnormalities (viz. stenotic atherosclerotic plaque, and saccular and fusiform aneurysmal lumens) in an indirect fashion, instead of directly manipulating the complex-shaped abnormalities. The proposed method has been tested on three synthetic and 17 clinical data sets. Comparisons with two related works are also conducted. Experimental results show that our method can produce satisfactory identification of the abnormalities and approximations of the ideal post-treatment vessel lumens. In addition, it can help increase the repeatability of the measurement of clinical parameters significantly.

Algorithms↗
PubMed1999

Technical note: a new model for quantitative analysis of brain oedema resolution into the ventricles and the subarachnoid space.

OBJECTIVE: The aim of the current study was to develop an experimental animal model for quantitative analysis of oedema resolution via the subarachnoid space and the ventricular system using fluorescent oedema markers. METHODS: Artificial cerebrospinal fluid (CSF) containing TRITC-albumin (MW 67.000D) and Na(+)-fluorescein (MW 376D) was continuously infused into the white matter of the left frontal lobe of New Zealand white rabbits (n = 6) at a rate of 100 microliters/h for 3 hrs. A closed cranial window for superfusion of the brain surface with artificial CSF fluid (3 ml/h) was implanted above the left parietal cortex for measurement of the fluorescence markers in the subarachnoid space. Uptake of the fluorescence indicators into the ventricles was quantified by ventriculo-cisternal perfusion (3 ml/h). The effluates were collected at 30 min intervals for 3 hrs after the start of infusion. Clearance of the oedema fluid into the perfusates was measured by fluorescence spectrophotometry. RESULTS: At an intracranial pressure of 15.0 +/- 1.7 mm Hg (mean +/- SEM) both indicators started to accumulate in the subarachnoid and ventricular perfusates at 90 min following onset of oedema fluid infusion. The concentrations of the indicators in the ventricular system increased to 7.7 +/- 5.1% of Na(+)-fluorescein and 16.1 +/- 13.0% of TRITC-albumin of the total amount infused were recovered in the ventricular system at 3 hours after start of the oedema infusion, while 3.4 +/- 3.2% of Na(+)-fluorescein and 3.7% +/- 3.2 of TRITC-albumin, respectively, were found in the effluates of the subarachnoid space. CONCLUSION: The present study demonstrates that resolution of vasogenic brain oedema into the cerebral ventricular system and the subarachnoid space following its entry into cerebral white matter can be quantitatively analysed using fluorescence markers, which serve as oedema fluid indicators. The results indicate that the oedema fluid is cleared not only into the ventricular system but also via the subarachnoid space.

Animals↗
PubMed1995

[Sex differences in EEG background activity: a study with quantitative analysis in normal adults].

The present study performed quantitative EEG analysis in normal 20 males and 20 females, aged 18-26 years, in order to examine the sex differences in EEG activity at rest. The females had more absolute EEG power than the males, with significant differences observed for delta, theta and beta bands. In contrast, no significant difference was found in peak alpha frequency or relative EEG power for any frequency band. The present findings provide further evidence that females have more absolute power in resting EEG, and emphasize the importance of matching patients and control populations for sex in future investigations dealing with the relationship between resting EEG and various pathological conditions.

Adolescent↗
PubMed1995

Quantitative analysis of THC, 11-OH-THC, and THCCOOH in human hair by negative ion chemical ionization mass spectrometry.

A sensitive and specific method was developed for the quantitative analysis of delta9-tetrahydrocannabinol (THC), 11 -hydroxy-THC (11-OH-THC), and 11-nor-9-carboxy-THC (THCCOOH) in human hair. Deuterated internal standards were added to hair samples, and samples were digested overnight in 1 N NaOH at 37 degrees C. Digest solutions were extracted with a liquid-liquid extraction procedure, which was previously developed in our laboratory for the analysis of plasma and whole blood. Derivatized extracts were analyzed on a Finnigan 4500" mass spectrometer in negative ion chemical ionization mode using methane as the reagent gas, hydrogen as the carrier gas, and a Restek Rtx 200-15M-0.25-microm capillary column. The assay was linear up to 50 ng/mg hair (r, 0.99) for all three compounds and was capable of detecting 10 pg THC and THCCOOH and 100 pg 11-OH-THC on column. The intra-assay precision ranged from 2.1 to 11.2% for the three analytes; the interassay precision ranged from 4.4 to 13.0%. The method was used to detect and quantitate the presence of THC, 11-OH-THC, and THCCOOH in human hair obtained from eight regular users of cannabis. THC, but not 11-OH-THC or THCCOOH, was detectable in the hair shaft above the assay limit of quantitation. Four laboratory wash procedures were also evaluated for their effect on the measured concentration of THC in hair. in seven of eight subjects, a methylene chloride wash procedure substantially reduced the measured THC concentration by up to 50%. The gas chromatographic-mass spectrometric assay is currently being used to support pharmacokinetic studies of drug disposition into the hair of humans and animals.

Boranes↗
PubMed2003

Flow injection-capillary electrophoresis system with contactless conductivity detection and hydrostatic pressure generated flow. Application to the quantitative analysis of inorganic anions in water samples.

A simple and inexpensive flow injection-capillary electrophoresis (FI-CE) system with contactless conductivity detection (CCD) for automated quantitative analysis of chloride, nitrate, and sulfate in various water samples is demonstrated. A glass bottle containing the background electrolyte that is raised above the FI-CE interface generates a pulse-free, highly reproducible flow of the electrolyte through the FI-CE interface. The system operates at a flow rate of 300 microLmin(-1) with an injection volume of only 4 microL. The repeatability of peak areas (n = 18) was better than 0.81% RSD and the sample throughput was 90 samples per hour using the background electrolyte containing 12 mM L-histidine adjusted to pH 4.00 with acetic acid. The limits of detection were better than 125 microgL(-1) and were comparable to those obtained by conventional CE systems with CCD. Various calibration methods for FI-CE system with electrokinetic injection were tested and their suitability for the analysis of anions in real samples was evaluated.

Anions↗
PubMed2004

Calibrationless quantitative analysis by indirect UV absorbance detection in capillary zone electrophoresis: the concept of the conversion factor.

A new, fast and efficient procedure is described for the simultaneous quantitative analysis of various non-UV absorbing species in a sample, by capillary zone electrophoresis with indirect UV absorbance detection. The procedure is based on the concept of the conversion factor (CF). The CF of an analyte is defined as the ratio of the measured temporal peak area and the product of its migration time and transfer ratio (TR). Thus defined, the CF is of general validity for all analytes separated and detected in a given background electrolyte (BGE), since it has the same value for the same amounts of various analytes. If a sample is enriched with a known concentration of a standard component and analyzed by CZE, the CF of this standard component can be calculated and then the concentrations of all other analytes can be determined, without the use of any calibration graph. The individual TRs can be determined a priori from tabulated ionic mobilities and pK values of the analytes and of the constituents of the BGE or, for strong analytes, by using experimental data from the electropherogram of the analysis itself. The practical procedure of the analysis includes enrichment of the sample with a known quantity of a suitable standard and a single CZE run of the resulting mixture. The injected volume does not need to be known and thus the procedure also eliminates the injection error. The proposed procedure has been verified experimentally and reproducible and accurate values were obtained by using four different CZE apparatus for the analyses of standard mixtures of cations in three different BGEs.

Calibration↗
PubMed2002

Description and validation of a staggered parallel high performance liquid chromatography system for good laboratory practice level quantitative analysis by liquid chromatography/tandem mass spectrometry.

The Aria LX4 staggered parallel high performance liquid chromatography (HPLC) system is evaluated for application to good laboratory practice (GLP) level quantitative analysis by liquid chromatography/tandem mass spectrometry (LC/MS/MS). This system consists of four fully independent binary HPLC pumps, a modified autosampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Validation results for four different compounds, each analyzed on a separate channel of the Aria system, show precision and accuracy equivalent to that required of a single-channel system. The results show that sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The flexibility and ease of use of the Aria system suggest that it should be possible to quickly implement it in any analytical LC/MS/MS environment.

Animals↗
PubMed2005

Quantitative analysis of the microbial metabolome by isotope dilution mass spectrometry using uniformly 13C-labeled cell extracts as internal standards.

A novel method was developed for the quantitative analysis of the microbial metabolome using a mixture of fully uniformly (U) (13)C-labeled metabolites as internal standard (IS) in the metabolite extraction procedure the subsequent liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis. This mixture of fully U (13)C-labeled metabolites was extracted from biomass of Saccharomyces cerevisiae cultivated in a fed-batch fermentation on fully U (13)C-labeled substrates. The obtained labeled cell extract contained, in principle, the whole yeast metabolome, allowing the quantification of any intracellular metabolite of interest in S. cerevisiae. We have applied the labeled cell extract as IS in the analysis of glycolytic and tricarboxylic acid (TCA) cycle intermediates in S. cerevisiae sampled in both steady-state and transient conditions following a glucose pulse. The use of labeled IS effectively reduced errors due to variations occurring in the analysis and sample processing. As a result, the linearity of calibration lines and the precision of measurements were significantly improved. Coextraction of the labeled cell extract with the samples also eliminates the need to perform elaborate recovery checks for each metabolite to be analyzed. In conclusion, the method presented leads to less workload, more robustness, and a higher precision in metabolome analysis.

Carbon Isotopes↗
PubMed2005

Multiple solid-phase microextraction in a non-equilibrium situation. Application in quantitative analysis of chlorophenols and chloroanisoles related to cork taint in wine.

Multiple HS-solid-phase microextraction (MHS-SPME) is a modification of SPME developed for quantitative analysis that avoids possible matrix effects based on an exhaustive analyte extraction from the sample. In this paper, the theory of this process associated with a non-equilibrium situation has been presented. The application of an optimised HS-SPME-based method in the analysis of chloroanisoles and chlorophenols, previously acetylated, associated with the occurrence of cork taint in different red, white and rosé wine samples, has revealed the existence of matrix effects. This fact determines the choice of standard addition as the adequate technique for the quantification of these compounds in real samples. MHS-SPME is proposed as a good alternative technique with respect to HS-SPME because it avoids matrix effects, simplifies the quantification of these compounds in real samples and reduces analysis time, providing sensitivity below chloroanisole sensory threshold with acceptable precision.

Anisoles↗
PubMed1997

Single point dilution method for the quantitative analysis of antibodies to the gag24 protein of HIV-1.

In the present work a concept proposed in 1992 by Dopotka and Giesendorf was applied to the quantitative analysis of antibodies to the p24 protein of HIV-1 in infected asymptomatic individuals and AIDS patients. Two approaches were analyzed, a linear model OD = b0 + b1.log(titer) and a nonlinear log(titer) = alpha.OD beta, similar to the Dopotka-Giesendorf's model. The above two proposed models adequately fit the dependence of the optical density values at a single point dilution, and titers achieved by the end point dilution method (EPDM). Nevertheless, the nonlinear model better fits the experimental data, according to residuals analysis. Classical EPDM was compared with the new single point dilution method (SPDM) using both models. The best correlation between titers calculated using both models and titers achieved by EPDM was obtained with the nonlinear model. The correlation coefficients for the nonlinear and linear models were r = 0.85 and r = 0.77, respectively. A new correction factor was introduced into the nonlinear model and this reduced the day-to-day variation of titer values. In general, SPDM saves time, reagents and is more precise and sensitive to changes in antibody levels, and therefore has a higher resolution than EPDM.

Enzyme-Linked Immunosorbent Assay↗
PubMed2000

Quantitative analysis of flavan-3-ols in Spanish foodstuffs and beverages.

An HPLC method, using detection after postcolumn derivatization with p-dimethylaminocynnamaldehyde (DMACA), was developed for the quantitative analysis of individual flavanols in food. This method was applied to flavanol determination in 56 different kinds of Spanish food products, including fruit, vegetables, legumes, beverages (cider, coffee, beer, tea, and wine), and chocolate. The determined compounds corresponded to the catechins and proanthocyanidin dimers and trimers usually present in food and, therefore, they were representative of the flavanols of low degree of polymerization consumed with the diet. The data generated could be used for calculation of the dietary intake of either individual or total flavanols, which would allow the further establishment of epidemiological correlations with the incidence of chronic diseases. Similar flavanol profiles were found in the different samples of a similar type of product, even though important variations could exist in the concentrations of total and individual flavanols among them. This was attributed to factors such as sample origin, stage of ripeness, post-harvesting conservation, and processing. Total flavanol contents varied from nondetectable in most of the vegetables to 184 mg/100 g found in a sample of broad bean. Substantial amounts were also found in some fruits, such as plum and apple, as well as in tea and red wine. Epicatechin was the most abundant flavanol, followed by catechin and procyanidin B2. In general, catechins were found in all the flavanol-containing products, but the presence of gallocatechins was only relevant in pomegranate, broad bean, lentil, grape, wine, beer, and tea, and most of the berries. Galloyled flavanols were only detected in strawberry, medlar, grape, and tea.

Beverages↗
PubMed1989

Quantitative analysis of systolic blood pressure tracking during childhood and adolescence using a tracking index: the Shimane Heart Study.

We used a tracking index to carry out a quantitative analysis of blood pressure tracking in children and adolescents. The index was calculated according to changes in blood pressure quintiles for the whole population during the observation period. The study population consisted of 463 Japanese children living in Izumo, who were examined every 3 years from 6 to 12 years of age in cohort 1, and from 9 to 15 years of age in cohort 2. In both sexes, the tracking index decreased transiently during the period from 9 to 12 years of age and increased again thereafter. The index was higher in girls than in boys except for the period from 12 to 15 years. Correlation coefficients between blood pressure at the first and that at the second examination increased with age in both sexes. We conclude that the tracking index could quantify the degree of tracking. Systolic blood pressure tracked well during childhood and adolescence, and the degree of tracking increased after the age of 12 years in both sexes.

Adolescent↗
PubMed2001

Quantitative analysis of estrogen receptor mRNA in human endometrium throughout the menstrual cycle using a real-time reverse transcription-polymerase chain reaction assay.

We conducted a quantitative analysis of ERalpha and ERbeta mRNA expression in normal human endometrium throughout the menstrual cycle in regular menstruating premenopausal women, taking advantage of this real-time PCR assay. Endometrial dating was determined from the histology of the endometrium and classified into: proliferative endometrium and secretory endometrium. Both ERalpha and ERbeta mRNA expression were detected in all endometrial samples at both proliferative and secretion phase. However ERalpha mRNA expression level was higher than that of ERbeta specially during proliferative phase. These results suggest that estrogenic effects occur predominantly through ERalpha than ERbeta.

Adult↗
PubMed2002

Quantitative analysis of the structure of the human extraocular muscle pulley system.

PURPOSE: Extraocular muscle (EOM) paths are constrained by connective tissue pulleys serving as functional origins. The quantitative structural features of pulleys and their intercouplings and orbital suspensions remain undetermined. This study was designed to quantify the composition of EOM pulleys and suspensory tissues. METHODS: Five human orbits, ages 33 weeks gestation to 93 years, were imaged intact by magnetic resonance (MRI), serially sectioned at 10 micro m thickness, and stained for collagen, elastin, and smooth muscle (SM). With MRI used as a reference, digital images of sections were geometrically corrected for shrinkage and processing deformations, and normalized to standard normal adult globe diameter. EOM pulleys, interconnections, suspensory tissues, and entheses were quantitatively analyzed for collagen, elastin, and SM thickness and density. RESULTS: Rectus and inferior oblique pulleys had uniform structural features in all specimens, comprising a dense EOM encirclement by collagen 1 to 2 mm thick. Elastin distribution varied, but was greatest in the orbital suspension of the medial rectus pulley and in a band from it to the inferior rectus pulley. This region corresponded to maximum SM density. Structural features of pulleys, intercouplings, and entheses were similar among specimens. The major mechanical couplings to the osseous orbit were near the medial and lateral rectus pulleys. CONCLUSIONS: Quantitative analysis of structure and composition of EOM pulleys and their suspensions is consistent with in vivo MRI observations showing discrete inflections in EOM paths that shift predictably with gaze. Focal SM distributions in the suspensions suggest distinct roles in stiffening as well as shifting rectus pulleys.

Aged↗
PubMed1998

Simultaneous quantitative analysis of prostaglandins and thromboxane after low-dose X irradiation.

The appearance of prostaglandins and thromboxane in mouse serum after X irradiation was observed by simultaneous quantitative analysis using gas chromatography/mass spectrometry/selected ion monitoring with stable isotope dilution methods. Mice of two strains (C57BL/CN Jcl and BALB/c) showed similar responses to X irradiation. In C57BL/6N Jcl mice, 0.2 Gy irradiation elicited a significant increase in generation of prostanoids: Immediately after irradiation, the 6-keto PGF1 alpha:TXB2 ratio and the level of PGE2 increased, after 20 min 6-keto PGF1 alpha and PGE2 increased, and after 4 h PGE1 and PGE2 increased. In BALB/c mice, generation of prostanoids was increased significantly immediately after irradiation (6-keto PGF1 alpha, 6-keto PGF1 alpha:TXB2 ratio, PGE2), and the increase was maintained from 20 min to 4 h (PGE1, PGE2) after 0.2 Gy irradiation. In C57BL/6N Jcl mice, a significant increase in production of 9alpha,11beta-PGF2 was observed at 20 min after irradiation. In BALB/c mice, a significant increase in 9alpha,11beta-PGF2 was seen immediately after irradiation and was maintained for 20 min. In C57BL/6N Jcl mice, the level of 8-epi PGF2 alpha was clearly increased 4 h after 4 Gy irradiation. A slight and slow increase was also seen after 0.2 Gy irradiation. In BALB/c mice, 8-epi PGF2 alpha was increased significantly at 20 min and 4 h after 4 Gy irradiation. These results show that 0.2 Gy irradiation stimulates production of prostanoids related to the inflammatory response in mice.

Animals↗
PubMed2001

Quantitative analysis of estrogen receptor-alpha and -beta messenger RNA expression in normal and malignant thyroid tissues by real-time polymerase chain reaction.

OBJECTIVES: We have conducted a quantitative analysis of estrogen receptor-alpha (ER-alpha) and -beta (ER-beta) mRNA expression in normal thyroid and tumor tissues. METHODS: Normal thyroid tissues (n = 10) and tumor tissues [(follicular adenoma (n = 14), follicular carcinoma (n = 8), papillary carcinoma (n = 14), anaplastic carcinoma (n = 3) and medullary carcinoma (n = 6)] were obtained at surgery from 45 female patients. ER-alpha and ER-beta mRNA expression has been studied by a quantitative polymerase chain reaction. RESULTS: ER-alpha mRNA levels in the normal thyroid were not significantly different from those in follicular adenomas, papillary carcinomas and medullary carcinomas but were marginally (p = 0.08) higher than those in follicular and anaplastic carcinomas. ER-beta mRNA levels in the normal thyroid tissues were not significantly different from those in any other tumor tissues. ER-beta to ER-alpha mRNA ratios were significantly (p < 0.05) higher in the normal thyroid tissues than in follicular adenomas. Proportions of ER-beta mRNA variants (ER-beta 1, 2, 5, and 5') did not significantly differ among the normal and tumor tissues. CONCLUSIONS: A downregulation of ER-alpha mRNA in follicular and anaplastic carcinomas seems to suggest that estrogens are unlikely to play an important role in the carcinogenesis and progression of these carcinomas. On the other hand, a significant decrease in ER-beta to ER-alpha mRNA ratios in follicular adenomas suggests a possible involvement of estrogens in the pathogenesis of this disease since the same phenomenon has been reported on estrogen-dependent breast cancers.

Adenocarcinoma, Follicular↗
PubMed1994

Quantitative analysis of bencynonate in human plasma using a deuterated internal standard by gas chromatography-mass spectrometry with selected-ion monitoring.

A gas chromatographic-mass spectrometric method is described for the quantitative analysis of bencynonate in human plasma. Deuterated bencynonate served as the internal standard and selected-ion monitoring of the fragments of bencynonate and internal standard permitted the quantitation of bencynonate down to 25 pg/ml of plasma. The assay is linear for plasma bencynonate concentrations in the range 25 pg/ml-3 ng/ml. At 0.25 ng/ml the recovery and coefficient of variation are 54.3% and 19.1%, respectively. Application of the method to clinical studies gave data for the pharmacokinetics and relative bioavailability of bencynonate in man.

Animals↗