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At least 955 records · Page 53Linked to original sources
PubMed1997

A quantitative analysis of the glial cell reaction in primary sensory termination areas following sciatic nerve injury and treatment with nerve growth factor in the adult rat.

The time course of the astroglial cell reaction in the nucleus gracilis and the spinal cord dorsal horn was examined following sciatic nerve transection in the adult rat with qualitative and quantitative analysis of glial fibrillary acidic protein immunoreactivity and in situ hybridization for its mRNA. In addition, the potential effect of exogenous nerve growth factor (NGF) was examined on the astroglial and microglial cells in the spinal cord dorsal horn at certain time points following sciatic nerve transection. An increase in glial fibrillary acidic protein immunoreactivity as well as mRNA labelling was observed from 1 day after lesioning, with a peak at about 1 week and 2 days after lesioning, respectively, followed by a decline. However, NGF application during 1, 2 and 4 weeks following nerve transection did not result in any significantly reduced astroglial or microglial activity. Our results show that the astroglial cell response in the nucleus gracilis and the spinal cord dorsal horn is rapid in comparison with previously described central degenerative changes following peripheral nerve lesions (transganglionic degeneration), that the astroglial cell reaction develops concomitantly with the microglial cell reaction previously described and that the "signal" from the axotomized neurons which induces these reactions can not be prevented by exogenous NGF applied to the peripheral nerve.

Animals↗
PubMed2001

Quantitative analysis of hypertrophy in cardiac chambers in cyanotic tetralogy of Fallot.

Although early total corrective repair for cyanotic tetralogy of Fallot is now safely performed at many institutions, long-term complications after surgical repair have been demonstrated. Therefore, the optimal procedure and timing for surgical treatment remain controxersial. In the present study, we conducted a quantitative analysis of the hypertrophy of all four chambers of 87 autopsied hearts of cyanotic tetralogy of Fallot and 71 normal control hearts utilizing the myocardial mass index, and evaluated the progression of lesions with advancing age. In cyanotic tetralogy of Fallot, hypertrophy of the right ventricle progresses immediately after birth, with that of the right atrium developing soon after. The left side of the heart is normal or slightly atrophied which could be corrected by sufficient palliative intervention or total corrective repair. The growth curves of both ventricles were parallel to those of normal hearts for the period studied. Pulmonary atresia, palliative operation, and total corrective repair have been shown to have some influence on the morphological characteristics of hearts of cyanotic tetralogy of Fallot.

Child, Preschool↗
PubMed2003

[Quantitative analysis of p53 and related genes mRNA in rat hepatocarcinogenesis induced by 3'-Me-DAB].

BACKGROUND & OBJECTIVE: p53 gene mutations and abnormal expression of p53 in hepatocarcinoma have been reported, but alteration in mRNA level is not yet understood. In order to find out the alteration in mRNA levels of p53, glutathione S-transferase P (GST-P), alpha-fetoprotein (AFP), and albumin in genesis, developing, and prognosis of hepatocarcinoma, quantitative analysis of mRNA levels of p53, GST-P, AFP, and albumin in prehepatocarcinoma and hepatocarcinoma foci was performed. METHODS: During hepatocarcinogenesis of F344 rat induced by 3'-methyl-4-dimethylamino-azobenzene (3'-Me-DAB), the levels of these mRNAs were quantitatively analyzed by LightCycler V3 System real-time RT-PCR after capturing accurately micro-foci in prehepatocarcinoma and hepato-carcinoma of rats with laser capture microdissection (LCM). RESULTS: At the 6th, 12th and 24th experiment weeks, the p53 mRNA levels in all of prehepatocarcinoma foci were markedly higher than those in the adjacent normal tissues (all of P < or = 0.001), and gradually decreased from the 6th week to the 24th week (P < 0.01). The content of p53 mRNA in hepatocarcinoma foci was higher than that in normal tissue (P = 0.028 and 0.013), but lower than that in prehepatocarcinoma foci. At 24th weeks, the sections of livers exhibited intensive immunostaining of p53 protein in prehepatocarcinoma and hepatocarcinoma foci. At any time-point of experiment, GST-P mRNA levels in prehepatocarcinoma foci were significantly higher than those in the adjacent normal liver tissue and hepatocarcinoma foci (all of P < 0.001). The concentration of AFP mRNA was the highest (P < 0.001) and that of albumin mRNA was the lowest (P < 0.01) in hepatocarcinoma foci as compared with adjacent normal tissue and prehepatocarcinoma foci. The GST-P protein and AFP protein were expressed strongly in prehepatocarcinoma and hepatocarcinoma foci, respectively. CONCLUSION: GST-P and AFP mRNA overexpressed in prehepatocarcinoma and hepatocarcinoma foci respectively will be profitable markers for diagnosis during hepatocarcinogenesis. The p53 mRNA highly expressed at early stage of prehepatocarcinoma and hepatocarcinoma, but the increasing concentration of p53 protein was found at later stage.

Actins↗
PubMed1990

Quantitative analysis of Tn10 Tet repressor binding to a complete set of tet operator mutants.

A saturating oligonucleotide-directed mutagenesis of both tet operators in the tet regulatory sequence was performed yielding mutants with four identical base pair exchanges at equivalent positions in the four tet operator half sides. The mutants were cloned between bipolar lacZ and galK indicator genes on a multicopy plasmid allowing the quantitative analysis of their effects in vivo. In the absence of Tet repressor the mutations lead to considerably different expression levels of both genes. They are discussed with respect to the promoter consensus sequences. In particular, the -10 region of the in vivo active tetPR2 promoter is unambiguously defined by these results. In the presence of Tet repressor most of the mutants exhibit a lower affinity for that protein as determined quantitatively by their reduced expression levels. In general, tet operator recognition is most strongly affected by alterations of base pairs near the center of the palindromic sequence. The most important position is the third base pair, followed by base pairs two, four, five and six, the latter showing similar effects as base pair one. At each position, the four possible base pairs show different affinities for Tet repressor. They are discussed according to their exposure of H-bond donors and -acceptors in the major and minor grooves of the B-DNA. The results are in agreement with major groove contacts at positions two, three and five. At position four a low potential correlation of efficiencies with the H-bonding in the minor groove is found, while mutations at position six seem to influence repressor binding by other mechanisms.

Bacterial Proteins↗
PubMed1996

Quantitative analysis of pancreatic glucokinase gene expression in cultured beta cells by competitive polymerase chain reaction.

Regulation of glucokinase (GK) gene expression in pancreatic beta cells has been poorly investigated, both due to low abundance of the gene and to difficulties in cells isolation. The present study describes the establishment of a competitive RT-PCR method for quantitative analysis of GK gene. The method has been applied to the analysis of GK mRNA expression RIN 1046-38 cells. We have monitored modifications of GK mRNA expression after different periods of time in culture and we have studied the effect induced by dexamethasone (DEX) treatment. We show that the method is very sensitive and requires very low amount of RNA. Data demonstrate that GK mRNA expression in RIN cells is reduced as a function of passages in culture and that the reduction is positively correlated with the decrease of insulin responsiveness observed in high passages cells. DEX treatment inhibits GK mRNA expression in RIN cells in a dose-dependent and time-dependent manner.

Animals↗
PubMedDate not supplied

[A quantitative analysis of complex formation between IgM and immobilized ligand using atomic force microscopy].

Specific interaction between human IgM and polyclonal antibodies immobilized on support was studied by atomic force microscopy. Human IgMs are responsible for a number of side effects arising during the xenotransplantation of mammalian organs to man. On the basis of atomic force microscopy, a quantitative analysis of complexes with IgM was performed. The data of the analysis agree well with the results of enzyme immunoassay. It was shown that the method of detection of immune complexes based on atomic force microscopy is able to detect specific antibodies/antigens in serum.

Aluminum Silicates↗
PubMed1992

[Quantitative analysis of urinary organic acid].

We showed the optimum conditions for analysis of urinary organic acids using solvent extraction and GC/MS by the stable isotope dilution technique. Examples of the diagnosis in a case of heterozygote of ornithine transcarbamylase deficiency and a case of multiple carboxylase deficiency were described. The accurate quantitative analysis of urinary organic acids with stable isotope dilution technique is necessary for routine examination conducted in clinical laboratories.

Female↗
PubMed2004

Automated quantitative analysis of HDM2 expression in malignant melanoma shows association with early-stage disease and improved outcome.

The incidence of cutaneous malignant melanoma continues to increase every year, and this disease remains the leading cause of skin cancer death in industrialized countries. Despite the aggressive nature of advanced melanoma, there are no standard biological assays in clinical usage that can predict metastasis. This may be due, in part, to the inadequacy of reproducible assessment of protein expression using traditional immunohistochemistry. We have previously described a novel method of quantitative assessment of protein expression (AQUA) with the continuity and accuracy of an ELISA assay but with maintenance of critical spatial information. Here, we modify this technology for the evaluation of protein expression in melanoma. Using a tissue microarray cohort of 405 melanoma lesions and 17 normal skin samples, we analyzed expression of HDM2, the human homologue of murine double minute 2 with automated quantitative analysis. We show that expression levels in the nucleus are significantly higher in primary melanomas than in metastatic lesions. Furthermore, high levels of expression are predictive of better outcome. This study demonstrates that quantitative assessment of protein expression is useful in melanoma to validate potential tissue biomarkers and suggests that human homologue of murine double minute 2 may be a valuable prognostic tool for management of malignant melanoma.

Biomarkers, Tumor↗
PubMed1982

The role of androgens in follicular development in the ovary. I. A quantitative analysis of oocyte ovulation.

In an attempt to understand more fully processes that control the selection or recruitment of follicles for ovulation, the superovulation paradigm in combination with the androgen, 5 alpha-dihydrotestosterone, or the antiandrogens, cyproterone or cyproterone acetate, was used in the immature mouse to alter the developmental potential of follicles destined to ovulate or to become atretic. Quantitative analysis of the numbers of eggs ovulated after one or more rounds of stimulation by pregnant mare's serum gonadotrophin followed by human chorionic gonadotrophin (PMSG-hCG), revealed a dose-dependent ovulation response to exogenous androgen and antiandrogen. Low dosages of androgen improved the ovulation response significantly. Large dosages of cyproterone and cyproterone acetate (100 mg/kg body weight) generally decreased the ovulation number in gonadotrophin-injected mice, suggesting a role for androgen in preovulatory events that occur within the ripened follicle after the ovulatory stimulus (hCG) has been received. Low dosages of cyproterone, particularly 25 mg/kg, significantly enhanced the ovulatory response, a phenomenon not observed for cyproterone acetate at this dosage. Radioimmunoassays of serum LH suggested that the differential response of the ovary to the two antiandrogens was probably related to endogenous LH release. Experiments in which the time of administration of hCG +/- cyproterone was varied after PMSG priming suggested that cyproterone at a dosage of 25 mg/kg had a "rescuing" effect on follicles destined to become atretic for up to 96 hr after PMSG priming. Cyproterone at a dosage of 100 mg/kg had no such effect, and actually decreased the magnitude of the ovulatory response at all time points tested, suggesting that follicular atresia was accelerated by this treatment. Experiments in which the time of administration of cyproterone (100 mg/kg) was varied after hCG suggested that whatever the important androgen-mediated events preceding ovulation, these events occur within 2 to 3 hr after the hCG signal. By quantitating the numbers of eggs over several superovulation cycles, it could be shown that hormonal treatment in one induced cycle could affect significantly the ovulation response in subsequent cycles, suggesting that androgens influence the development of classes of follicles other than preovulatory follicles. These studies suggest that the process through which follicles are selected for ovulation is extremely sensitive to the androgenic environment and that the developmental pathways leading to ovulation or preovulatory follicular atresia are closely linked.

Animals↗
PubMedDate not supplied

Method for quantitative analysis of glycosaminoglycan distribution in cultured natural and engineered cartilage.

Cartilage tissue engineering can provide a valuable tool for controlled studies of tissue development. As an example, analysis of the spatial distribution of glycosaminoglycans (GAG) in sections of cartilaginous tissues engineered under different culture conditions could be used to correlate the effects of environmental factors with the structure of the regenerated tissue. In this paper we describe a computer-based technique for quantitative analysis of safranin-O stained histological sections, using low magnification light microscopy images. We identified a parameter to quantify the intensity of red color in the sections, which in turn was proportional to the biochemically determined wet weight fraction of GAG in corresponding tissue samples, and to describe the spatial distribution of GAG as a function of depth from the section edge. A broken line regression model was then used to determine the thickness of an external region, with lower GAG fractions, and the spatial rate of change in GAG content. The method was applied to the quantitation of GAG distribution in samples of natural and engineered cartilage, cultured for 6 weeks in three different vessels: static flasks, mixed flasks, and rotating bioreactors.

Animals↗
PubMed1980

A quantitative analysis of ethanol and acetaldehyde expired by inbred mouse strains.

Expired ethanol and acetaldehyde were measured after an oral injection of ethanol in C57BL/6J and DBA/2J mouse strains by a combination of several techniques in a sequence involving a method for trapping expired radioactive compounds, separation of compounds by gas chromatography, isolation of radioactive ethanol and acetaldehyde, and their quantitative analysis by liquid scintillation spectrophotometry. With the specific activities used in evaluation of the technique (0.1 Ci/mole, acetaldehyde; 1.1 Ci/mol, ethanol) the lower limit of sensitivity using 500 microliters from a 10 ml trap is 955 pmoles for acetaldehyde and 101 pmoles for ethanol. However, in the animal experiments, injected ethanol has a specific activity of 1.1 Ci/mol which would make the specific activity of expired metabolically formed acetaldehyde the same. This results in a lower limit of sensitivity for acetaldehyde of 80 pmoles. The two strains were monitored for 80 min following an oral injection of 3.8 g/Kg of (2-14C) ethanol. Comparing the two strains on the expiration of each compound the curves were identical.

Acetaldehyde↗
PubMed2005

A pattern recognition tool for quantitative analysis of in planta hyphal growth of powdery mildew fungi.

The development of fungal pathogens can be quantified easily at the level of spore germination or penetration. However, the exact quantification of hyphal growth rates after initial, successful host invasion is much more difficult. Here, we report on the development of a new pattern recognition software (HyphArea) for automated quantitative analysis of hyphal growth rates of powdery mildew fungi on plant surfaces that usually represent highly irregular and noisy image backgrounds. By using HyphArea, we measured growth rates of colonies of the barley powdery mildew, Blumeria graminis f. sp. hordei, on susceptible and induced-resistant host plants. Hyphal growth was not influenced by the resistance state of the plants up to 48 h postinoculation. At later time points, growth rate increased on susceptible plants, whereas it remained restricted on induced-resistant plants. This difference in hyphal growth rate was accompanied by lack of secondary haustoria formation on induced-resistant plants, suggesting that induced resistance in barley against Blumeria graminis is caused mainly by reduced penetration rates of primary as well as secondary appressoria leading, finally, to fewer and less-developed fungal colonies. No evidence was found for reduced nutrient-uptake efficiency of the primary haustoria in induced-resistant leaves, which would be expected to have resulted in reduced hyphal growth rates during the first 48 h of the interaction.

Ascomycota↗
PubMed1981

Segmental quantitative analysis of digital thallium-201 myocardial scintigrams in diagnosis of coronary artery disease. Comparison with rest and exercise electrocardiography and coronary arteriography.

One hundred and forty-nine patients with suspected ischaemic heart disease were evaluated by exercise thallium-201 myocardial scintigraphy (201Tl SMS), single lead exercise electrocardiography, and coronary arteriography. Myocardial distribution of tracer was assessed semi-quantitatively from digital 201Tl scintigrams and compared with tracer distribution in subjects with normal hearts. Fifty-two of 54 (96%) patients with normal coronary arteries had normal myocardial scintigrams whereas three patients had a positive ischaemic exercise electrocardiogram and were scan normal. Conversely, 36 of 95 (38%) patients with coronary artery disease had a positive ischaemic electrocardiogram compared with 94 of 95 (99%) patients who had a positive myocardial scintigram. Disease in specific coronary arteries could be deduced from the topography of myocardial tracer deficit. Disease was predicted correctly in 76 out of 80 (95%) of left anterior descending coronary stenoses, in 48 out of 64 (75%) of right coronary artery stenoses, and in 55 out of 64 (85%) of left circumflex coronary artery stenosis, despite the presence of infarcted myocardium in other territories. Similarly, single vessel disease was predicted correctly in 14 out of 17 (82%) patients and multiple vessel disease in 64 out of 77 (83%) patients. 201Tl SMS with segmental quantitative analysis is a highly sensitive and specific technique in the diagnosis of coronary artery disease and may be useful screening procedure to select patients for further investigation, particularly those with evidence of life-threatening severe left coronary artery disease.

Adult↗
PubMed1988

[Treadmill exercise echocardiography: quantitative analysis of regional left ventricular wall motion by computer graphics].

To detect significant coronary lesions based on exercise-induced reversible asynergy, two-dimensional echocardiograms were recorded before and immediately after treadmill exercise test in 15 patients with angina pectoris (AP) and six patients with neurocirculatory asthenia (NCA). Short-axis views of the left ventricle were analyzed quantitatively, using the following indices: segmental area change (%A); segmental wall thickness change (%Th); and relative curvature (rC) which was the product of curvature of each segment multiplied by end-diastolic circumference. The results were compared with those of coronary angiography, T1-201 myocardial emission computed tomography, and exercise electrocardiography. 1. The results of comparisons of quantitative analysis by %A, %Th, and rC with qualitative (visual) analysis were as follows: the sensitivities were 57%, 74% and 91%; the specificities were 75%, 75% and 90%; and the accuracies were 68%, 75% and 90%, respectively. 2. To detect coronary stenosis of more than 75%, visual assessments and assessments by rC were superior to assessments by %Th or %A. The accuracies were 93%, 91%, 73% and 61%, respectively. 3. In five cases with three-vessel disease, all diseased coronary arteries were detected by treadmill exercise echocardiography using rC as an index. However, by treadmill exercise T1-201 myocardial emission computed tomography, the diagnosis of three-vessel disease was possible in only one case. 4. The results of treadmill exercise electrocardiography were positive in 11 of 15 the AP patients and in all six NCA patients. The results of exercise echocardiography using rC as an index were normal in all NCA patients and abnormal in 14 of the 15 AP patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1996

Quantitative analysis of oligonucleotides by matrix-assisted laser desorption/ionization mass spectrometry.

Quantitative aspects of oligonucleotide analysis by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry remain largely unexplored relative to the efforts that have been devoted to quantitative peptide and protein analysis. The successful quantitation of these other biopolymers coupled with the potential of rapid nucleic acid analysis by desorption/ionization techniques prompted the present investigation into quantifying mixed base oligonucleotides of intermediate molecular weights. This report describes the concentration-dependent desorption/ionization of a 21-base oligonucleotide (MW 6361) using a 36-base oligonucleotide (MW 11 131) as an internal standard. Peak height and peak area ratios (analyte to internal standard) varied linearly as a function of oligonucleotide concentration (R2 = 0.966 and 0.991, respectively). The linearity of response extended over nearly three orders of magnitude, from 0.125 to 100 pmol of analyte applied. The use of an internal standard improved the linearity of the calibration curve and reduced relative standard deviations. These results demonstrate for the first time the quantitation of medium size oligonucleotides using MALDI.

Calibration↗
PubMed1998

Quantitative analysis of synaptophysin immunoreactivity in human neocortex after cardiac arrest: confocal laser scanning microscopy study.

Transient global ischaemia caused by cardiac arrest results in lesions that involve all brain structures. The aim of this study was to investigate the condition of synapses in patients surviving, but remaining in a persistent vegetative state, following resuscitation after cardiac arrest. We performed a quantitative analysis of the distribution and density of elements containing a synaptic vesicle protein--synaptophysin (p38)--in human neocortex in cases which survived for 1 week, 2 months, and 1 year after the cardiac arrest. Neurologically healthy cases that died following an accident served as control. Dual-channel confocal laser scanning microscopy (CLSM) was used to image p38-immunoreactivity (IR) and lipofuscin autofluorescence. In control cases no statistically significant differences were found for p38-IR between layers II-III and V-VII. However, the temporal cortex had a higher density of p38-immunoreactive structures than the motor cortex. In postischaemic cases a reduction in the density of p38-IR elements was apparent, mainly in the frontal and motor cortices and less pronounced in the temporal cortex. The least decrease compared with controls was observed in the visual cortex. In the 1 week survival case, a maximal decrease in p38-IR (35% below control) was found. In this case, the number of p38-IR elements per visual field was decreased, and big aggregates of p38-IR structures were observed. In general, the amounts of p38-IR structures were higher in all of the control cases compared with the postischaemic cases.

Autopsy↗
PubMed1986

Quantitative analysis of retinal outer segments in ovine ceroid-lipofuscinosis.

Isolated outer segments from the photoreceptors of the eye were markedly reduced in young animals with ovine ceroid-lipofuscinosis. At 5 months of age, 68% of these structures were lost and by 18 months, no segments remained. Our results suggest that loss of these structures can be identified earlier by quantitative analysis, than by electroretinography.

Animals↗
PubMed2002

Quantitative analysis of type IV collagen alpha chains in the basement membrane of human urogenital epithelium.

Type IV collagen is a major component of the basement membrane (BM), which consists of six genetically distinct alpha(IV) chains. In this study the expression of these six alpha(IV) chains was demonstrated immunohistochemically. In addition, the alpha2(IV) and alpha5(IV) chains were analysed quantitatively by confocal laser scanning microscopy in human urogenital epithelial BM. The alpha1/alpha2(IV) and alpha5/alpha6(IV) chains were immunoreactive in the epithelial BM, whereas, alpha3/alpha4(IV) chains were not. The quantitative analysis revealed that the amount of alpha2(IV) and alpha5(IV) chains differed in each urogenital epithelial BM. The content of alpha5(IV) chains in the epithelial BM of the bladder was differentially high, and that of the foreskin was differentially low. It is concluded that the elasticity of epithelial BM of the bladder may be structurally related to the high content of alpha5/alpha6(IV) chains.

Antibodies, Monoclonal↗