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PubMed1999

A quantitative analysis of oligodendrocytes in multiple sclerosis lesions. A study of 113 cases.

We studied quantitatively the fate of oligodendrocytes (OGs) during lesion formation in 395 lesion areas from biopsy and autopsy tissue of 113 multiple sclerosis cases. The density of OGs in multiple sclerosis lesions was variable at all stages of demyelinating activity, ranging from nearly complete loss to values exceeding those in the periplaque white matter (range 0-970 OGs/mm2) determine whether there were distinct patterns of OG pathology in different patients, we restricted our analysis to the 56 cases in which the longitudinal extent of the lesion extended from periplaque white matter into the active demyelinating edge and inactive plaque centre. Two major groups of OG pathology were defined by the presence or absence of increased OGs within the lesion. In 70% (39 out of 56) of the cases, OGs were variably reduced during active stages of myelin destruction, but reappeared within inactive or remyelinating areas. In inactive areas, an increased number of OGs expressing proteolipid protein (PLP) mRNA compared with those expressing myelin oligodendrocyte glycoprotein (MOG) suggested these cells may have been derived from the progenitor pool. In the remaining 30% (17 out of 56) of the cases, extensive destruction of myelinating cells at active sites of demyelination was observed, but OGs were absent in inactive plaque areas without remyelination. In all lesions from a given patient the pattern of OG pathology remained consistent. A highly significant negative correlation was observed between number of macrophages in lesions and number of MOG- and PLP mRNA-labelled OGs (MOG: r = -0.32, P < 0.0000118; PLP mRNA: r = -0.23, P < 0.00238). OG density did not correlate with T-cell and plasma cell inflammation, or axonal loss. The profound heterogeneity in extent and topography of OG destruction in active demyelinating lesions suggests that in subsets of multiple sclerosis patients, myelin, mature OGs and possibly OG progenitors are differentially affected.

Adult↗

PubMed1998

Quantitative analysis of ultrasound B-mode images of carotid atherosclerotic plaque: correlation with visual classification and histological examination.

This paper presents a quantitative comparison of three types of information available for 52 patients scheduled for carotid endarterectomy: subjective classification of the ultrasound images obtained during scanning before operation, first-and second-order statistical features extracted from regions of the plaque in still ultrasound images from three orthogonal scan planes and finally a histological analysis of the surgically removed plaque. The quantitative comparison was made with the linear model and with separation of the available data into training and test sets. The comparison of subjective classification with features from still ultrasound images revealed an overall agreement of 60% for classification of echogenicity and 70% for classification of structure. Comparison of the histologically determined relative volume of soft materials with features from the still images revealed a correlation coefficient of r = -0.42(p = 0.002). for mean echogenicity of the plaque region. The best performing feature was of second order and denoted Contrast (r = -0.5). Though significant, the latter correlation is probably not strong enough to be useful for clinical prediction of relative volume of soft materials for individual patients. Reasons for this is discussed in the paper, together with suggestions for improvements.

Arteriosclerosis↗

PubMed1993

[Quantitative analysis of brain perfusion using radionuclide angiography with 99mTc-HMPAO].

A noninvasive simple method for quantitative radionuclide angiography with 99mTc-dl-hexamethyl propylene amine oxime (99mTc-HMPAO). The method of graphical analysis was employed for the evaluation of the unidirectional influx constant (Ku) from the blood to the brain for the tracer. The Ku values were standardized to represent objective and comparable values, brain perfusion indices (BPI), among studied subjects by setting the ratio of ROIbrain size to ROIaorta size to 10. The mean +/- SD of the whole-brain BPI values for the 41 normal controls was 11.15 +/- 2.61, and biphasic reduction with advancing age was observed. The mean of the whole-brain BPI of 7.95 with SD of 2.35 in 103 patients with cerebrovascular disorders was significantly lower than that in normal controls. The right to left ratio of BPI showed wider dynamic range than that of the total counts of SPECT. This technique is quite easy to apply as an adjunct to SPECT and may be helpful in the quantitative evaluation of brain perfusion in routine clinical studies.

Adult↗

PubMed1995

Quantitative analysis of electrotonic structure and membrane properties of NMDA-activated lamprey spinal neurons.

Parameter optimization methods were used to quantitatively analyze frequency-domain-voltage-clamp data of NMDA-activated lamprey spinal neurons simultaneously over a wide range of membrane potentials. A neuronal cable model was used to explicitly take into account receptors located on the dendritic trees. The driving point membrane admittance was measured from the cell soma in response to a Fourier synthesized point voltage clamp stimulus. The data were fitted to an equivalent cable model consisting of a single lumped soma compartment coupled resistively to a series of equal dendritic compartments. The model contains voltage-dependent NMDA sensitive (INMDA), slow potassium (IK), and leakage (IL) currents. Both the passive cable properties and the voltage dependence of ion channel kinetics were estimated, including the electronic structure of the cell, the steady-state gating characteristics, and the time constants for particular voltage- and time-dependent ionic conductances. An alternate kinetic formulation was developed that consisted of steady-state values for the gating parameters and their time constants at half-activation values as well as slopes of these parameters at half-activation. This procedure allowed independent restrictions on the magnitude and slope of both the steady-state gating variable and its associated time constant. Quantitative estimates of the voltage-dependent membrane ion conductances and their kinetic parameters were used to solve the nonlinear equations describing dynamic responses. The model accurately predicts current clamp responses and is consistent with experimentally measured TTX-resistant NMDA-induced patterned activity. In summary, an analysis method is developed that provides a pragmatic approach to quantitatively describe a nonlinear neuronal system.

Animals↗

PubMed1998

Quantitative analysis of collagen fibrils of human cruciate and meniscofemoral ligaments.

The ultrastructural anatomy of collagen fibril diameters in the cruciate and meniscofemoral ligaments, from four young human cadaver knees (mean age, 20 years, range, 17-22 years) was studied using transmission electron microscopy. Samples were harvested from the proximal, middle, and distal regions of the anterior and posterior cruciate ligaments, and the meniscofemoral ligament. Photomicrographs were taken and assessed quantitatively using image analysis software to determine the collagen fibril diameters and eccentricities, and the percentage of total cross sectional area occupied by collagen. The collagen fibril diameter for the anterior cruciate ligament was found to be largest in the distal region but it decreased as it moved proximally. The posterior cruciate ligament had an opposite trend because it decreased from the proximal to the distal region. For the meniscofemoral ligament, the fibrils of the middle region were larger than those of the proximal and distal regions. The percentage of total cross sectional area occupied by collagen, however, did not vary significantly between regions. Fibril eccentricity also varied little between ligament or location. The variability observed in fibril diameters may account for the different mechanical properties of the ligaments.

Adolescent↗

PubMed2005

Optical metal ion sensor based on diffusion followed by an immobilizing reaction. Quantitative analysis by a mesoporous monolith containing functional groups.

A new optical metal ion sensor based on diffusion followed by an immobilizing reaction has been developed. The current sensor is based on a model that unifies two fundamental processes which a metal analyte undergoes when it is exposed to a porous, ligand-grafted monolith: (a) diffusion of metal ions to the binding sites and (b) metal-ligand (ML(n)) complexation. A slow diffusion of the metal ions is followed by their fast immobilizing reaction with the ligands in the monolith to give a complex. Inside the region where the ligands have been saturated, the diffusion of the metal ions reaches a steady state with a constant external metal ion concentration (C(0)). If the complex ML(n) could be observed spectroscopically, the absorbance of the product A(p) follows: A(p) = Kt(1/2), K = 2epsilon(p)(L(0)C(0)D)(1/2). D = diffusion constant of the metal ions inside the porous solid; L(0) = concentration of the ligands grafted in the monolith; and t = time. This equation is straightforward to use, and the K vs C(0)(1/2) plot provides the correlations with the concentrations (C(0)) of the metal ions. This is a rare optical sensor for quantitative metal ion analysis. The use of the model in a mesoporous sol-gel monolith containing grafted amine ligands for quantitative Cu(2+) sensing is demonstrated. This model may also be used in other chemical sensors that depend on diffusion of analytes followed by immobilizing reactions in porous sensors containing grafted/encapsulated functional groups/molecules.

Journal Article↗

PubMed2005

Quantitative analysis of gene regulation by seven clinically relevant progestins suggests a highly similar mechanism of action through progesterone receptors in T47D breast cancer cells.

Progesterone (P4) is an essential reproductive steroid hormone required for many aspects of female reproductive physiology. Progestins are compounds that demonstrate progesterone-like activity and are used in oral contraception, hormone therapy, and treatment of some reproductive disorders, but differ widely in their chemical structures, potency, and pharmacokinetics. While numerous studies have assessed progestins on specific endpoints, little is known about the activation of global gene expression by progestins. We used Affymetrix GeneChip U133A expression arrays to examine the action of P4 and six clinically relevant synthetic progestins (3-ketodesogestrel, drospirenone, levonorgestrel, medroxyprogesterone acetate, norethindrone acetate, and trimegestone) on the progesterone receptor (PR)-positive T47Dco and the PR-negative T47D-Y breast cancer cell lines. Excluding drospirenone, one or more of the progestins-regulated 329 genes, with 30 genes regulated by at least 2.0-fold by all progestins in the T47Dco cells. The synthetic progestins show a high degree of similarity in their transcriptional responses, and each progestin regulates between 77 and 91% of the genes regulated by P4. Independent quantitative RT-PCR analysis confirmed a similar regulation for S100P, PPL, IL20RA, NET1, ATP1A1, HIG2, and CXCL12 (SDF-1) by all seven progestins. Attempts to find differentially regulated genes by any progestin compared to all other treatments failed, suggesting any differences are quantitative, not qualitative. This analysis demonstrates a high degree of similarity among these progestins on PR-regulated gene expression in T47D cells, suggesting a similar and fairly specific mode of action.

Alkaline Phosphatase↗

PubMed2001

Quantitative analysis of chemokine expression by dendritic cell subsets in vitro and in vivo.

Upon maturation, dendritic cells (DCs) have to adjust their chemokine expression to sequentially attract different leukocyte subsets. We used real-time quantitative polymerase chain reaction analysis to study in detail the expression of 12 chemokines involved in the recruitment of leukocytes into and inside secondary lymphoid organs, by DCs in distinct differentiation stages, both in vitro and in vivo. Monocyte-derived immature DCs expressed high levels of DC chemokine 1 (DC-CK1), EBI1-ligand chemokine (ELC), macrophage-derived chemokine (MDC), macrophage-inflammatory protein (MIP)-1alpha, and thymus and activation-regulated chemokine (TARC). Upon maturation, DCs up-regulated the expression of DC-CK1 (60-fold), ELC (7-fold), and TARC (10-fold). Activation of DCs by CD40 ligand further up-regulated the expression of ELC (25-fold). We found that freshly isolated blood DCs expressed only low levels of interleukin-8, lymphotactin, and MIP-1alpha. It is interesting that the chemokine profile expressed by activated CD11c(-) lymphoid-like as well as CD11c(+) myeloid blood DCs mimics that of monocyte-derived DCS: Additionally, purified Langerhans cells that had migrated out of the epidermis expressed a similar chemokine pattern. These data indicate that different DC subsets in vitro and in vivo can express the same chemokines to attract leukocytes.

CD40 Ligand↗

PubMed1976

[Quantitative analysis of ovalbumin by thin-layer isoelectric focusing (author's transl)].

A new method for quantitation of ovalbumin by thin layer isoelectric focusing in polyacrylamide gel is described. This technique separates ovalbumin from avian-magnum homogenates or egg white and allows the quantitative determination of each ovalbumin fraction. Considering the very small amount (7 mul) of sample needed for an analysis, the sensitivity and the reproducibility of the proposed method are good. The application of this technique to quail egg white ovalbumin analysis leads to the characterization of four fractions A3, A2, A1 and A0 containing, respectively, 0, 1, 2 and 3 atoms of phosphorus per mole and with isoelectric points that range from 4.95 (A3) to 4.65 (A0).

Animals↗

PubMed2004

Qualitative and quantitative analysis of bacterial aerosols.

The objective of this study was to investigate qualitatively and quantitatively the bacterial aerosols before, during, and after clinical work sessions in different areas within a multichair dental clinic, an isolation clinic, the sterilization center, and the prosthetic laboratory in the College of Dentistry, King Saud University. Also, the contributions of aerosols generated by different types of dental procedures were investigated. Air sampling using blood and heart infusion agar plates at four selected areas was performed three times per day over a 2-week period before, during, and after clinical sessions. The concentration of total bacterial aerosols was 5 times higher in the multichair clinic, 3.6 times higher in the prosthetic laboratory, 2 times higher in the sterilization center and isolation clinic during working sessions as compared to before the working sessions. At the end of the working day, aerosols decreased 50-70% in all areas. Staphylococcus epidermidis had the highest prevalence (37.12q%) of colony composition of bacteria examined. This study demonstrates that aerosols increase during and after work sessions and, therefore, increase the chance for infectious agent transmission. Preventive measures should be instituted to reduce or disrupt aerosols as a transmission route in the multichair dental clinic, sterilization center, prosthetic laboratory, and isolation clinic.

Aerosols↗

PubMed2006

Quantitative analysis of chimerism after allogeneic stem cell transplantation by real-time polymerase chain reaction with single nucleotide polymorphisms, standard tandem repeats, and Y-chromosome-specific sequences.

We compared the results of chimerism analyses with real-time SNP-PCR to those obtained by the classical STR-PCR method in 135 hematopoietic stem cell transplantation recipients. Using 10 different SNP gene loci, the SNP-PCR method was able to discriminate patient from donor cells in 125 of 135 cases (93%), whereas the use of 11 different STR gene loci with the STR-PCR analysis using agarose or polyacrylamide gel resolution resulted in accurate donor-host discrimination in all patients. Of the 470 analyzed samples we found in 74% concordant results for both chimerism methods. In all 26% discordant cases the SNP-chimerism method showed mixed chimerism (MC), whereas the STR-method found complete chimerism (CC). As a consequence, the SNP-PCR chimerism analysis method detected a MC prior to the occurrence of relapse significantly earlier than the STR-PCR chimerism method (120 vs. 30 days, P < 0.007). The probability of relapses was significantly higher in patients with increasing MC (70%) compared to 30% in patients with CC (P < 0.00001) associated with a significantly shorter overall survival in patients with increasing MC. The multivariate Cox model showed that chimerism analsis with SNP-PCR was the only significant risk factor predicting relapse (RR 6.08, P < 0.0001).Furthermore, we analyzed the chimerism status in male recipients with a female donor in 580 samples of 134 patients using quantitative real-time PCR of Y-chromosome-specific sequences and compared the results with interphase XY-fluorescent in situ hybridization (FISH). MC without signs of relapse was detected in 35% of samples using quantitative real-time PCR of Y-chromosome-specific sequences. The detected Y-DNA amounts were low compared to the amounts detected in 104 samples of 42 patients with leukemic relapse at the time of analysis (P < 0.0001). Quantitative real-time PCR of Y-chromosome-specific sequences detected therefore an increasing MC with high residual host DNA amounts approximately 143 days (mean) prior to the occurrence of relapse. By comparing the results of Y-chromosome PCR with the XY-FISH analysis we found concordant results in 73% in patients with myeloablative regimens. The XY-FISH could detect 12 relapses, whereas the Y-chromosome PCR detect 36 relapses by MC (P < 0.005). Residual host cells gradually decreased during the posttransplant period from a mean of 5.4 ng (first months) to 0.5 ng (above 5 years) without evidence of relapses. The probability of relapses was significantly higher in patients with increasing MC (100%) compared to 8% in patients with CC (P < 0.00001) associated with a significantly shorter overall survival in patients with increasing MC. The multivariate Cox model showed that chimerism analysis of Y-chromosome-specific sequences is an important risk factor for relapse (RR 17.0, P < 0.0001). We conclude that the use of real-time SNP or Y-PCR may be superior to the STR-PCR or interphase XY-FISH methods in detecting patients who are at high risk for relapse after transplant.

Adolescent↗

PubMed1978

Quantitative analysis of propranolol and metabolites by a gas chromatograph mass spectrometer computer technique.

A quantitative assay for urinary propranolol and six of its unconjugated metabolites by a gas chromatography mass spectrometry computer system has been developed. Conjugated metabolites are also determined following enzyme hydrolysis. These compounds are separated from murine by Amberlite XAD-2 resin column chromatography and are converted to methyl esters and trifluoroacetyl derivatives. Each substance is quantitated from reconstructed ion chromatograms using a specific mass or sum of masses for each compound by comparing its area with that of an appropriate internal standard. The minimum detectable concentrations are in the range of 50--100 ng ml-1. The calibrations are linear from 0.10--15.0 microgram ml-1 with recoveries ranging from 62--91%. This procedure yields a high degree of specificity, selectivity and reproducibility, as demonstrated by the analyses of urine from dogs receiving single 40 mg doses of propranolol orally.

Animals↗

PubMed1984

Torpedo electromotor system development: a quantitative analysis of synaptogenesis.

Synaptogenesis in the electric organ of Torpedo marmorato has been studied quantitatively at the ultrastructural level of observation. In addition to establishing the normal developmental time course for this event we were interested in determining whether a gradient of synaptogenesis might be present because the electric organ produces several morphologically recognizable spatiotemporal gradients during its early ontogeny. These gradients genesis of electrocyte columns, both gradients of which are operative for periods of weeks. No gradient of synaptogenesis was found, indicating this to be a synchronous process. The idea is advanced that synaptogenesis in the electric organ is modulated by extrinsic influences, many of which may originate from the target electrocytes which, by this time, have become synchronized in their development.

Acetylcholinesterase↗

PubMed1991

A method for the quantitative analysis and standardization of interleukin-1 bioactivity.

A method is presented for the reproducible quantitation of the biological activity of interleukin 1 (IL-1). This method provides diagnostic tools which give insights into the qualitative aspects of the binding of IL-1 and of the resulting activation of the responder thymocytes; for example, whether the lymphokine and/or the responder population is heterogeneous, or whether a threshold level exists. It establishes under what circumstances the assumptions on which it is based are reasonably adhered to and, consequently, quantitative estimation in the manner it prescribes is justified. It also gives a simple way to calculate both the maximal response attainable for each preparation in an assay and the dilution of a particular preparation that would produce a half-maximal response, the accepted unit of activity of IL-1. This empirical technique provides an improved means of comparing the activities of various preparations of IL-1 in bioassays using various stocks of responder cells and reagents. It should also be applicable to the evaluation of the biological activity of lymphokines in general.

Animals↗

PubMed1983

A computerized method of quantitative analysis of ventricular phase distributions.

Phase images from gated equilibrium cardiac studies, were analysed quantitatively by a computerized automatic method. The study included 17 normal subjects and 33 patients with an acute myocardial infarction (AMI). Phase histograms were constructed for each ventricle and their peaks were identified by a convolution algorithm for smoothing and differentiation. Intra- and interventricular phase shifts as well as the percentage of normal contracting pixels (NCP) were calculated. The results showed that the left ventricular ejection fraction (LVEF) tends to decrease when the intraventricular phase shift increases. Interventricular phase shifts out of the normal range (+/- 10 degrees) were observed predominantly in patients with an extensive anterior or anteroseptal AMI. The closest relationship was found between LVEF and the percentage of NCP. This parameter decreases when akinetic and dyskinetic areas increase and could be considered as a quantitative index of the synchronism in myocardial contraction useful in the prognosis and follow-up of patients.

Adolescent↗

PubMed2004

Quantitative analysis of immobilized metalloenzymes by atomic absorption spectroscopy.

A new, sensitive assay for the quantitative determination of immobilized metal containing enzymes has been developed using atomic absorption spectroscopy (AAS). In contrast with conventionally used indirect methods the described quantitative AAS assay for metalloenzymes allows more exact analyses, because the carrier material with the enzyme is investigated directly. As an example, the validity and reliability of the method was examined by fixing the iron-containing enzyme catalase on cotton fabrics using different immobilization techniques. Sample preparation was carried out by dissolving the loaded fabrics in sulfuric acid before oxidising the residues with hydrogen peroxide. The iron concentrations were determined by flame atomic absorption spectrometry after calibration of the spectrometer with solutions of the free enzyme at different concentrations.

Catalase↗

PubMed2005

Quantitative analysis of protein-protein interactions by native page/fluorimaging.

We have developed a new quantitative native PAGE mobility shift assay, which allows for the measurement of binding affinities for interacting protein pairs, one of which is fluorescently labelled. We have used it to examine recognition of the Simian virus 40 (SV40) large tumour T-antigen (T-ag) nuclear localisation sequence (NLS) by members of the importin (Imp) superfamily of nuclear transport proteins. We demonstrate that the T-ag NLS binds to the Imp alpha/beta heterodimer in NLS-dependent manner, determining that it binds with eight-fold higher affinity (340 nM), when compared to Imp alpha alone, consistent with autoinhibition of Imp alpha when not complexed with Imp beta. The mobility shift assay is able to detect nM binding affinities, making it a sensitive and useful tool to analyse protein-protein interactions in solution.

Animals↗

PubMed1988

Quantitative analysis of internalization of glucagon by isolated hepatocytes.

Biochemical methods have been used to quantitate total, acid-stable and acid-labile association of (mono[125I]iodoTyr10) glucagon with rat hepatocytes in suspension to evaluate internalization of glucagon and its receptors. Internalization is inhibited by low temperature, phenylarsine oxide, and by blocking receptor binding, consistent with receptor-mediated endocytosis. Approximately 30% of the total cell-associated hormone is internalized at 30 min of incubation. The rate declines until 90 min when the internalization of glucagon ceases, although the cells remain competent to internalize asialofetuin. From 90 min to 4 h, 27% of the maximum label internalized at 30 min remains within cells. The number of cell surface receptors decreases but the affinity of those remaining is unchanged. However, 1.7-2.7 surface receptors are lost to binding for each molecule of radiolabeled glucagon internalized. Uptake occurs according to a rate constant of 0.183 min-1 (t1/2 = 3.8 min). We conclude that (i) hepatocytes internalize a finite quantity of glucagon, implying the existence of undefined regulatory mechanisms; (ii) hormone is retained for greater than 2 h within cells and may play a physiological role within cells; and (iii) both occupied and unoccupied receptors become inaccessible to extracellular hormone as internalization proceeds; rapid recycling of receptors does not occur.

Animals↗