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[Quantitative analysis of EEG basic rhythms in epileptics (author's transl)].

Basic rhythms of EEG were quantitatively analyzed in 92 adult idiopathic epileptic patients and in 57 age-matched control subjects. Epileptic patients have been administered one or both of anticonvulsants, phenobarbital (PB) and diphenyl-hydantoin (DPH). Before the EEG study, blood samples were taken from patients and serum concentration of PB and DPH was examined with UV method. Dose and serum level correlation was statistically significant in both drugs but fairly inferior in DPH. Dominant posterior rhythms in awake subjects were restored in a medical computer. These data were transformed with a Fast Fourier method into amplitude spectra, which were expressed in percentage distribution of each frequency band, i.e. delta (1.0--3.5 Hz), theta (4.0--7.5), alpha (8.0--12.5) and beta activity (13.0--29.5 Hz). As the first result, it was found that slow wave activity significantly increased and alpha activity decreased in epileptics compared with control subjects. No difference was found in beta activity. So as the next step, the etiologies of abnormal slowness of epileptic EEG were investigated in relation to serum level of anticonvulsants, age of seizure onset, seizure frequency, duration of medication and seizure types. These analysis confirmed that slowing of epileptic EEG has statistically significant correlation with the following factors, 1) high serum level of anticonvulsants (PB greater than 20 microgram/ml), DPH greater than 15 microgram/ml) 2) high frequency of seizures (more than once per month) 3) seizure onset at young ages (0--5 yrs.) 4) psychomotor seizure type. When these slowing factors are plurally involved in one patient, the degree of EEG slowness increased in proportion to the numbers of these factors. On the other hand, if the comparison was made between epileptics without these factors and control subjects, strikingly no minimum differences was found in basic rhythms. This result induces the very important conclusion that epilepsy itself has no direct effect on slowing of basics rhythms and if there exists slowing, some secondary reasons must be searched. The author also speculated pathogenetic causes of EEG slowing by these factors and insisted that there exist very close interrelation between basic EEG rhythms and prognosis of epilepsy.

Adult↗

Quantitative analysis of gait pattern in hemiparetic patients.

OBJECTIVE: To characterise gait pattern in hemiparetic patients quantitatively using clinical footprint method. DESIGN: A case control study. SUBJECTS: Sixteen hemiparetic patients (12 males and 4 females) aged 16 to 64 years who attended neurological clinic at Queen Elizabeth Central Hospital, Blantyre, Malawi. MAIN OUTCOME MEASURES: Stride length, step width, foot rotation angle measured using footprint method. RESULTS: The difference in mean values of the three variables of gait between affected and unaffected sides in hemiparetic patients was not significant. However, strides were significantly longer in controls than in patients (P<0.001) while patients had significantly broader steps (P<0.02). In the controls, foot was rotated externally in 87.7% of steps and internally in 14.3% of steps. In the hemiparetic patients, intoeing pattern was observed in 41.3% of steps. The difference in variability of stride length and foot angle between affected and non-affected sides of patients was not significant. Stride-to-stride variability in stride length in patients was 1.6 times higher than in controls. Variability of step width and foot progression angle was 1.4 and 1.6 times higher in patients than in controls. CONCLUSION: Footprint method provides fast and inexpensive tool for clinical gait analysis and is suitable for evaluation of hemiparetic patients. Our findings suggest that areas of emphasis for physical therapy of hemiparetic patients should include increasing stride length and decreasing step width and internal rotation of foot, particularly on the affected side.

Adolescent↗

The value of quantitative analysis of glucose utilization in detection of myocardial viability by PET.

To study whether absolute quantitation of regional myocardial glucose utilization (rMGU) enhances detection of myocardial viability, 70 nondiabetic patients with prior myocardial infarction and angiographically confirmed coronary artery disease were studied with [18F]FDG PET after oral glucose loading. Forty-eight patients were also revascularized and underwent echocardiography after revascularization to detect wall motion recovery. The rMGU was calculated in eight myocardial segments in each patient and the results were compared to normalized (relative) [18F]FDG uptake values. In normal segments (n = 225), rMGU was 56 +/- 18 mumole/min/100 g (mean +/- s.d.) and relative [18F]FDG uptake 97% +/- 12%. The interindividual variation of rMGU in normal myocardium was greater than the intraindividual variation (s.d. 31% versus 11%). The respective values for relative [18F]FDG uptake were 9% and 10%. Both rMGU and [18F]FDG uptake were significantly reduced in segments with scarring observed visually during bypass surgery (29 +/- 19 mumole/min/100 g and 45% +/- 22%, n = 26). The rMGU and [18F]FDG uptake were higher in segments that recovered after revascularization (53 +/- 17 mumole/min/100/g and 110% +/- 21%, n = 27) than in those that did not (37 +/- 20 mumole/min/100 g and 65% +/- 24%, n = 63). However, due to larger variability of rMGU values, normalized [18F]FDG uptake was superior to rMGU in separating normal and scar segments as well as in predicting wall motion recovery. We conclude that rMGU variability is notable and is caused mainly by variations between patients. Interindividual variation is reduced by normalization, which results in more accurate assessment of myocardial viability. Thus, static imaging and semiquantitative analysis are sufficient for the clinical assessment of myocardial viability.

Coronary Angiography↗

Quantitative analysis of herpes simplex virus in cranial nerve ganglia.

A susceptible individual exposed to herpes simplex virus (HSV) will develop latent infection in multiple cranial nerve ganglia. There are a few quantitative studies of the viral load within the trigeminal ganglion, but none that investigate other cranial nerve ganglia. In this study, human trigeminal, geniculate, vestibular (Scarpa's) and cochlear (spiral) ganglia were obtained from willed body donors. Real time quantitative polymerase chain reaction (PCR) analysis of the HSV DNA polymerase gene was performed on ipsilateral ganglion sets from the same individual. Viral load, expressed as HSV genomes per 105 cells, was significantly greater in the vestibular ganglion (mean +/- SD, 176705 +/- 255916) than in the geniculate (9948 +/- 22066), cochlear (3527 +/- 9360), or trigeminal (2017 +/- 5578) ganglia. There was not a significant correlation among ganglia from the same individual. The results support the hypothesis that neuronal subpopulations have variable susceptibility to HSV infection.

Base Sequence↗

Quantitative analysis of alpha 1 (I) procollagen transcripts in vivo by competitive polymerase chain reaction.

Due to the limited amount of RNA obtainable from punch biopsies, few data exist on the human alpha 1 (I) procollagen mRNA steady state level in vivo. Therefore, we established a competitive PCR method to quantitate this mRNA in human biopsies. In our approach, the target template and the standard share the same sequence except for a 69 bp deletion, thus competing for the same primer pair and subsequently amplifying at the same rate. Titration of a competitor DNA dilution series against an unknown cDNA sample then allows quantitation of the separated and nonradioactively detected PCR products after densitometrical analysis. Almost identical results were obtained by quantitations of the same total RNA by competitive PCR as well as Northern blot analysis.

Base Sequence↗

Quantitative analysis of the behavior of Dictyostelium discoideum amoebae: stringency of pteridine reception.

A convenient, sensitive, quantitative assay for the measurement of chemotaxis of populations of D. discoideum vegetative amoebae is presented. A strategy for determining the boundary of the bulk of a population of migrating amoebae was devised and is described. This assay employs a dynamic gradient and is independent of deaminase activity. Measurements of chemoattractant capabilities of various pteridines, folates, and mixtures of folate fragments are reported. 2-Amino 4-quinazolinone, a pterin analog without the pyrazine ring nitrogens, is chemotactic. Lumazine, deaminated pterin, inhibits chemotaxis towards pterin but not towards folic acid. Deaminofolic acid is a chemoattractant as are mixtures of lumazine plus aminobenzoylglutamic acid or deaminopteroic acid plus various amino acids. Separately, the components of these mixtures exhibit no ability to stimulate chemotaxis. These mixtures are of fragments that together comprise most of the folate structure. Our results are in accord with separate receptors for pterin vs. folic acid and with a high stringency for pterin reception but a relative tolerance for folate reception. The possibility of using such mixtures to investigate the requirements of various parts of the folate structure for competent signalling is discussed.

Animals↗

Quantitative analysis of XG blood group and CD99 antigens on human red cells.

CD99, the product of the MIC2 gene, exhibits an erythroid-specific quantitative polymorphism co-regulated with the Xga blood group polymorphism. The co-expression of X-linked MIC2 and XG genes is presumably controlled at the transcriptional level by a single XGR locus in the pseudoautosomal region of sexual chromosomes. This locus is composed of two alleles, XGR(low) and XGR(high), which determine low or high CD99 levels (CD99-L, CD99-H) and the Xg(a-)/ Xg(a+) status. To test this hypothesis, the phenotypic relationship between Xga and CD99 antigens on human RBCs was investigated by quantitative flow cytometry using NBL-1 (anti-Xga) and 12E7 (anti-CD99) monoclonal antibodies and semi-quantitative estimate of membrane proteins and RNA by Western blot and Northern blot, respectively. The antibody binding capacity of RBCs, which is an estimation of the antigen density, was determined for 118 blood donors including 60 males and 58 females. Xg(a+) RBCs, which all belong to the group of CD99-H expressors, carry 159+/-13 and 960+/-50 copies of Xga and CD99 molecules/cell, respectively. Xg(a-) RBCs have no Xga antigen, but are subdivided into CD99-H (all male) and CD99-L expressors carrying 747+/-28 and 200+/-22 CD99 copies/cell, respectively, with identical CD99 levels between CD99-L males and females. However, among males, the CD99 expression was higher in Xg(a+) than in Xg(a-)/CD99-H individuals (P<0.01). In addition, CD99-H expressors in Xg(a+) males could be clearly subdivided into two categories, high and super high expressors, which are presumably heterozygous and homozygous for the XGR(high) allele, which fits the above hypothesis. This was not the case for Xg(a+) females where CD99-H subcategories were not found. Quantitative differences were confirmed by Western blot analysis of red cell membrane preparations from individuals of different Xga and CD99 phenotypes and by Northern blot analysis showing that the reticulocytes from CD99-L individuals expressed a reduced level of MIC2 transcripts compared to CD99-H donors. These findings further support the hypothesis of a single genetic control of CD99 and Xga expression by the XGR locus.

12E7 Antigen↗

A quantitative analysis of synaptogenesis in the molecular layer of the dentate gyrus in the rhesus monkey.

Quantitative electron microscopy was used to study synapse formation in the molecular layer of the dentate gyrus in rhesus monkeys ranging in age from embryonic day 62 to adult. Four to eight radial probes, consisting of a series of overlapping electronmicrographs and extending across the full thickness of the molecular layer were made in each specimen. Synaptic density (normalized to volume of neuropil) increased significantly during the last half of gestation, reaching adult levels at the time of birth. However, new synapses were added during infancy, resulting in an apparent peak in density at between 4 and 5 months of age. This increase was followed by a decline in the synaptic density over the next 5 months, to levels comparable to that of the newborn. In addition to synaptic density, synapse type (symmetric, asymmetric), location (on dendritic shafts or spines), and laminar distribution in the developing molecular layer was determined. The decrease in synaptic density is unlikely to be due to 'dilution' caused by an increase in molecular layer volume since no increase in the volume of the dentate gyrus could be detected during this period. Our calculations suggest that a selective overproduction of asymmetrical, axo-spinous synapses occurs during infancy. Finally, synaptic density was significantly greater in the middle third of the molecular layer suggesting that synaptic exuberance may be related to entorhinal input.

Aging↗

Rapid quantitative analysis of paraquat by electron spin resonance spectroscopy.

A very simple and sensitive method for quantitative measurement of paraquat in plasma, urine and some drinks is described. The paraquat radical formed by its reduction can be detected by electron spin resonance spectroscopy (ESR) at room temperature without clean-up or concentration process of samples. The sensitivity limit is 0.1 microgram paraquat/ml and the required amount of sample is 100 microliter. The time needed for the measurement is within 10 min.

Beverages↗

American tegumentary leishmaniasis: a quantitative analysis of Langerhans cells presents important differences between L. (L.) amazonensis and Viannia subgenus.

A quantitative study was conducted on the density of Langerhans cells (LCs) CD1a+ in specimens obtained from patients with American tegumentary leishmaniasis (ATL) lesions without previous treatment, as well as from control healthy individuals. LC density was significantly higher among infected patients when compared to controls and also higher in longer term ones. Regarding parasite quantities, these were proportionally inverse and diminished in chronic patients. Localized cutaneous leishmaniasis (LCL) showed an increase in cell population when compared to diffuse cutaneous leishmaniasis (DCL). A tendency towards density increase was observed in LC Leishmania (Leishmania) amazonensis patients when compared to Leishmania (Viannia) sp. Regarding the delayed hypersensitivity test (DTH, Montenegro skin test), L. (L.) amazonensis demonstrated a peculiar behavior because it is a poor cell immune inducer, presenting--among LCL patients--higher density in negative Montenegro patients than in positive ones. Negative DTH responses are usually poor in LC, although this was not evidenced in this study, possibly due to cell reposition, in order to stimulate immune response. Such results confirm the important role of LC in ATL, while suggesting that L. (L.) amazonensis may be a good model for LC studies as APC in ATL, due to its spectral immunological and clinical behavior.

Adolescent↗

Direct observation and quantitative analysis of spatiotemporal dynamics of individual living monocytes during transendothelial migration.

OBJECTIVE: To visualize and quantitatively analyze spatiotemporal dynamics of individual living monocytes during transendothelial migration (TEM). METHODS AND RESULTS: We developed an in vitro new experimental system using confocal laser scanning microscope with following two improvements: (1) ultra thin collagen gel layer (30-50 microm thick) constructed under human umbilical vein endothelial cell layer for three-dimensional observation with high magnification; (2) appropriate fluorescent labeling of living monocytes and endothelial cells to keep highest cell activity. Individual monocytes behaved quite diversely. Approximately 70% of adhered monocytes directionally crawled to intercellular junction, and started invasion. Time from adhesion to start of invasion was 8.6 +/- 5.4 min (mean +/- S.D., n=61 monocytes). Approximately 80% of such invading monocytes completed TEM, but remaining 20% of once invading monocytes hesitated transmigration, and returned onto the endothelial surface. Time from start to finish of invasion was 6.3 +/- 3.2 min (mean +/- S.D., n=53 monocytes). CONCLUSIONS: Using our collagen gel-based newly-developed system, we visualized and quantitatively analyzed detailed spatiotemporal, three-dimensional dynamics of individual living monocytes during TEM. We revealed that monocytes encountered at least two hurdles, at starting invasion, and leaving endothelium, to achieve complete TEM. Approximately 56% (80% of 70% of adhered monocytes) passed both hurdles.

Cell Movement↗

Application of capillary zone electrophoresis and reversed-phase high-performance liquid chromatography in the biopharmaceutical industry for the quantitative analysis of the monosaccharides released from a highly glycosylated therapeutic protein.

Two assays for the quantitative determination of the neutral and amino-monosaccharides attached to a therapeutic glycoprotein were developed using capillary zone electrophoresis (CZE) and RP-HPLC. These assays meet the strict batch release requirements of the quality control in biopharmaceutical industry. The monosaccharides were released from the glycoprotein by hydrolysis with 2N trifluoroacetic acid. In the CZE assay the monosaccharides were reacetylated prior to derivatization with 8-aminopyrenesulfonic acid (APTS), reacetylation in the glycoprotein matrix was investigated in detail. The RP-HPLC method used pre-column derivatization with anthranilic acid in methanol-acetate-borate reaction medium; reacetylation was not necessary. However, epimerization of the different monosaccharides was observed and studied in detail. For the quantitative assay, separation of the amino-monosaccharide epimers had to be developed. The HPLC assay was validated.

Chromatography, High Pressure Liquid↗

Ion-exchange chromatography followed by ESI-MS for quantitative analysis of sugar monophosphates from glucose catabolism.

The aim of this work is to establish a quantitative method to determine the ratio of [U-(13)C] labeled to unlabeled hexose monophosphates isolated from yeast extracts. This is accomplished by anion exchange chromatography and mobile phase desalting followed by electrospray (ESI) mass spectrometry. We test the method with the analysis of a sample of biological origin. Previously developed analytical techniques are not adequate to accomplish mass spectrometric analysis of these and other small monosaccharide systems because of interference from salt clusters. By lowering the ionic strength of the mobile phase and using a simplified injection system to the mass spectrometer, we were able to obtain data on the relative abundance of the hexose monophosphates.

Carbohydrates↗

Improvement of the inverse-gated-decoupling sequence for a faster quantitative analysis of various samples by 13C NMR spectroscopy.

The inverse-gated-decoupling sequence enables quantitative (1)H decoupled (13)C spectra to be obtained. We modified this sequence so as to obtain the same result in less time for molecules containing carbons with various relaxation properties. For that, we determined the optimal (13)C longitudinal-magnetization initial value for a faster relaxation while (1)H decoupler is stopped. This value can be calculated precisely via the nuclear Overhauser effects, the longitudinal relaxation times, together with the determination of the relaxation rate constants of carbons while (1)H are out of equilibrium. A supplementary delay of (1)H decoupling and/or a series of selective pulses applied at the beginning of the recovery delay allow an acceleration of (13)C longitudinal relaxation. We applied this method to the molecule of vanillin. The simultaneous quantification of all carbons was carried out with a recovery delay divided by two compared to the usual sequence.

Benzaldehydes↗

Immunohistochemical study of the inflammatory infiltrate in villitis of unknown etiology. A qualitative and quantitative analysis.

Villitis is characterized by an inflammatory infiltrate within the substance of the chorionic villi. Quantitative and qualitative analyses of the mononuclear infiltrate in areas of villitis were performed in placentas with villitis of unknown etiology (VUE). We used a panel of monoclonal antibodies and immunoperoxidase technique in paraffin sections from 17 placentas with VUE and 8 without VUE. Macrophages followed by T lymphocytes were the predominant inflammatory cells in areas of villitis in virtually all cases. B lymphocytes were not observed and monocytes were present usually in small number in 58 per cent of the cases. Mononuclear cells which expressed HLA-DR antigens were found in 75 per cent of the cases. In areas of villitis with trophoblastic necrosis, we found monocytes and some T lymphocytes adhered to them. These cells apparently had migrated from the maternal circulation. We suggest that in areas of villitis with destruction of the trophoblast and its basal membrane the inflammatory infiltrate might have a mixture of fetal and maternal cells. The maternal monocytes and T lymphocytes might be attracted to these sites of trophoblastic necrosis and activated due to exposure to fetal MHC antigens of the villous stroma.

Chorionic Villi↗

Quantitative analysis of beta-phenylpyruvic acid by single ion monitoring. Evaluation of isomeric internal standards.

Quantitative single ion monitoring of beta-phenylpyruvic acid at high sensitivity is possible after derivatization first with omicron-phenylenediamine and then with a silylating reagent. The resulting o-trimethyl-silyl-quinoxalinol (O-TMS-Q) has previously been shown to be highly stable during storage and on chromatography. As an internal standard the isomeric omicron-methylphenylglyoxylic (omicron-toluylformic) acid is introduced. The mass spectra of both O-TMS-Q's are characterized by abundant [M]+. at m/e 308. The concept of "class specific metabolic profiling" is discussed in relation to quantitative gas chromatography--mass spectrometry detection of aliphatic and aromatic alpha-ketoacids.

Chromatography, Gas↗

Time-resolved NIR spectroscopy for quantitative analysis of intact pharmaceutical tablets.

Near-infrared (NIR) spectroscopy is a useful technique for quantitative measurements of intact tablets, but it suffers from limitations due to the fact that changes in the physical properties of a sample strongly affect the recorded spectrum. In this work, time-resolved transmission NIR spectroscopy was utilized to conduct quantitative measurements of intact tablets. The technique enables separation of the absorption properties of the sample from the scattering properties and can therefore handle changes of the physical parameters of the samples in a better way than conventional NIR transmission spectroscopy. The experiments were conducted using a pulsed Ti:sapphire laser coupled into a nonlinear photonic crystal fiber as light source. The light transmitted through the sample was measured by a time-resolving streak camera. A comparison of the results from the time-resolved technique with the results from conventional transmission NIR spectroscopy was made using tablets containing different concentrations of iron oxide and manufactured with different thicknesses. A PLS model made with data from the time-resolved technique predicted samples 5 times better than a PLS model made data from the conventional NIR transmission technique. Furthermore, an improvement to predict samples with physical properties outside those included in the calibration set was demonstrated.

Calibration↗