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Quantitative analysis of intracranial circulation using rapid-sequence DSA.

By using a high-frame-rate technique, intravenous digital subtraction angiography (IVDSA) of the intracranial circulation was performed in patients with transient ischemic attacks and asymptomatic bruits. Twenty patients with normal carotid arteries or mild stenosis of no hemodynamic significance were selected as a control group to evaluate the effect of carotid stenosis on the difference between hemispheres in the peak arrival time (delta TMAX) of the contrast bolus. Data were obtained for the anterior (ACA), middle (MCA), and posterior (PCA) cerebral artery distributions. Raw data of the time-density curve in a region of interest were analyzed by polynomial curve-fitting techniques to obtain the peak arrival time (TMAX). The delta TMAXs for normal middle, anterior, and posterior distributions were 0.140 +/- 0.119 sec, 0.152 +/- 0.146 sec, and 0.189 +/- 0.187 sec, respectively. Eleven patients with tight carotid stenosis or occlusion whose delta TMAX fell outside the normal range as established from the 20 control patients were analyzed with regard to ischemic symptoms. The delta TMAXs of the MCA, ACA distributions in the asymptomatic patients with tight carotid stenosis or occlusion were 0.653 +/- 0.379 sec and 0.118 +/- 0.159 sec, respectively; p less than 0.001 in the MCA when compared with controls. The delta TMAXs of the MCA, ACA distribution in the symptomatic patients with tight carotid stenosis or occlusion were 1.31 +/- 0.13 sec and 0.525 +/- 0.079 sec, respectively; p less than 0.001 in the MCA and ACA distributions when compared with controls. Quantitative analysis of this type of physiologic data may enable the detection of patients with carotid stenosis who are well compensated by collateral flow from those who are poorly compensated and at risk for possible infarction on a hemodynamic basis. Serial followup DSA studies in patients with asymptomatic bruits may help to correlate the progression of extracranial carotid stenosis and the status of intracranial collateral reserves.

Adult↗

Quantitative analysis of protein adsorption on a planar surface by Fourier transform infrared spectroscopy: lysozyme adsorbed on hydrophobic silicon-containing polymer.

The adsorption of hen egg white lysozyme onto a solid polytris(trimethylsiloxy)silylstyrene (pTSS) surface from a D(2)O solution at pD 7 containing 100 mM NaCl and 10 mM sodium deuterated phosphate was monitored at 25 degrees C by Fourier transform infrared spectroscopy using the attenuated total reflection (ATR) method. The infrared spectrum attributed to only the adsorbed lysozyme was derived from the observed spectrum, and the amount of adsorbed lysozyme was determined as a function of time and lysozyme concentration. The kinetics of adsorption could be decomposed into two components, one of which was a process with a time constant of larger than 4 h(-1) and the other was a process with one of about 0.1 h(-1). These spectra showed that the lysozyme adsorbed in the faster process had a higher beta-structure content than the dissolved lysozyme. It was also found that the slower adsorption induced some conformational change in the lysozyme adsorbed in the faster process and/or that adsorbed in the slower process. After adsorption for 24 h, the pTSS surface was rinsed out with lysozyme-free solution. The resultant spectra of the surface indicated that the lysozyme adsorbed in the faster process was bound irreversibly on the surface and was changed to a conformer with a higher beta-structure content during the slower process. The experimental procedures and the theoretical applications for such a quantitative analysis in the ATR spectroscopic method are presented in detail.

Adsorption↗

Real-time quantitative analysis for human telomerase reverse transcriptase mRNA and human telomerase RNA component mRNA expressions as markers for clinicopathologic parameters in urinary bladder cancer.

AIM: The expression of the telomerase subunits such as human telomerase reverse transcriptase (hTERT) and human telomerase RNA component (hTR) may be associated with tumor development and progression. We evaluated the relationship between mRNA quantification of both hTERT and hTR and clinicopathologic parameters in bladder cancer. METHODS: We examined the mRNA expression of hTERT and hTR in 29 specimens with bladder cancer (Grade: Grade I, 9 cases; Grade II, 13 cases and Grade III, 7 cases. Stage: pTa-pT1, 18 cases; pT2-T4, 11 cases). We immediately froze all of specimens obtained during TUR-Bt and isolated the total RNA from each specimen. We measured the quantity of hTERT, hTR and GAPDH mRNA by a real-time reverse transcription-polymerase chain reaction method based on TaqMan technology. RESULTS: The hTERT/GAPDH mRNA ratio and hTERT mRNA/total RNA in superficial bladder tumor was significantly lower than in invasive bladder tumor. The hTR/GAPDH mRNA ratio and hTR mRNA/total RNA in superficial tumor were significantly lower than in invasive bladder tumor. The hTERT mRNA expression significantly correlated with tumor grade, but the hTR mRNA expression did not correlate with tumor grade. There was no significant difference in the hTERT/GAPDH mRNA ratio and hTR mRNA/total RNA according to multiplicity of bladder tumor. CONCLUSIONS: Our results demonstrated that the expression of hTERT mRNA correlated with the progression of stage and grade in bladder cancer. The quantitative analysis of hTERT and hTR mRNA might be a marker for clinicopathologic parameters in bladder cancer.

Adult↗

Neonatal screening for galactosemia by quantitative analysis of hexose monophosphates using tandem mass spectrometry: a retrospective study.

BACKGROUND: Classic galactosemia (OMIM 230400) is an inherited disorder in the metabolism of galactose caused by deficiency of the enzyme galactose 1-phosphate uridyl transferase (EC 2.7.7.12). Galactosemia leads to accumulation of galactose and galactose 1-phosphate (gal-1-P) in blood and tissues and, if untreated, produces neonatal death or severe mental retardation, cirrhosis of the liver, and cataracts. Hence, the disorder is included in many neonatal screening programs. METHODS: We retrospectively analyzed filter-paper blood samples obtained 4-8 days postpartum for routine neonatal screening from 12 galactosemia patients and 2055 random controls. Total hexose monophosphates (HMPs) were used as a marker of gal-1-P and were assayed by negative-ion mode electrospray tandem mass spectrometry (tandem MS) with settings biased toward gal-1-P detection. The predominant precursor/product ion pair m/z 259/79 was used to quantify total HMPs by external standardization. RESULTS: Linear calibration curves were obtained in the range 0-8 mmol/L gal-1-P. The detection limit was 0.1 mmol/L HMP, and total CVs ranged from 13% at the detection limit to <8% at >1 mmol/L HMP. The method was in agreement with an alkaline phosphatase-galactose dehydrogenase method. All samples from galactosemia patients contained increased HMP concentrations (range for patients, 2.6-5.2 mmol/L; range for reference group, <0.10-0.94 mmol/L). The diagnostic sensitivity and specificity were 100% at a cutoff of 1.2 mmol/L HMP. A Duarte/classic galactosemia compound heterozygous sample could be discriminated clearly from both patient and reference samples. CONCLUSION: Quantitative analysis of HMPs by tandem MS can be used in laboratory investigations of galactosemia.

Alkaline Phosphatase↗

Quantitative analysis of analgoantipyretics in dosage form using planar chromatography.

In the therapy of pain of weaker genesis, frequently used drugs usually represent a mix of analgoantipyretics of different chemical structures, mostly derivatives of salicylic acid, pyrazolone and p-aminophenol as well as derivatives of propionic and acetylsalicylic acid. For the determination of these drugs, different chromatographic methods have been applied, mostly HPLC, due to the the lower polarity (pyrazolones derivatives) and thermolability, as well as nonvolatility of compounds investigated. TLC method, considering advantages which include simplicity, reasonable sensitivity, rapidity, excellent resolving power and low cost has been successfully explored for the determination of analgoantipyretic compounds. The aim of this work was to develop a simple and rapid HPTLC method for the determination of acetylsalicylic acid, paracetamol, caffeine and phenobarbitone in dosage form. The determination of analgoantipyretics were performed on pre-coated HPTLC silica gel plates (10 x 20 cm(2)) by development in the mobile phase dichlormethane-ethyl acetate-cyclohexane-isopropanol-0.1 M HCL-formic acid (9:8:3:1.5:0.2:0.2 v/v/v/v/v/v). Migration distances (68.6+0.2 mm, 54.1+0.1 mm, 36.4+0.14 mm and 85.9+0.11 mm for acetylsalicylic acid, paracetamol, caffeine and phenobarbitone, respectively) with low RSD values (0.13--0.39%) showed a satisfactory reproductivity of the chromatographic system. TLC scanner was used for direct evaluation of the chromatograms in the reflectance/absorbance mode. Established calibration curves (r>0.999), precision (0.3--1.02%) and detection limits, as well as recovery values (96.51--98.1%) were validated and found to be satisfactory. The method was found to be reproducible and convenient for the quantitative analysis of compounds investigated in their dosage forms.

Analgesics↗

Seriate histomorphometry of whole rat stomach: an accurate and reliable method for quantitative analysis of mucosal damage.

The evaluation of mucosal damage in experimental models of gastric injury is commonly based on macroscopic detection of gross lesions and/or histological examination of tissue samples and is limited by the subjectivity of the examiner and by the paucity of nonrepresentative samples. This study proposes a novel method for the histomorphometric analysis of gastric damage, based on the examination of seriate parallel strips taken from whole rat stomachs. Strips were cut perpendicular to the lesser curvature, placed on a glass slide, with the side of each strip distal to the pylorus upward, and processed for routine histology. Sections were then observed by light microscopy: the length of damaged mucosa divided by the total length of mucosa, measured on a micrometric scale and expressed in percentage values, was indicated as the lesion index. Furthermore, to evaluate the severity of the damage, three types of lesions were discriminated depending on their depth: type I, lysis of luminal cells; type II, damage involving the cells lying on both surface mucosa and gastric pits; and type III, damage involving the lower part of the lamina propria with injury of glands associated with detachment of whole mucosal layers. Three models of acute gastric damage (ethanol, hemorrhagic shock, and indomethacin) were examined and treatment was also carried out with the antiulcer drugs omeprazole, ranitidine, and misoprostol, to show the advantages of this histomorphometric approach. The results indicate that this method allows an accurate quantitative analysis of gastric damage, and the effects of different antiulcer drugs can be better discriminated.

Animals↗

Quantitative analysis of liposome-cell interactions in vitro: rate constants of binding and endocytosis with suspension and adherent J774 cells and human monocytes.

We have characterized the parameters describing the total association (uptake) of liposomes with murine macrophage-like cell line J774 cells and human peripheral blood monocytes at 4 degrees C and at 37 degrees C with or without inhibitors of endocytosis. The uptake of neutral liposomes composed of phosphatidylcholine (PC)/cholesterol (Chol) (2:1 mole ratio) is about 10-fold lower than that of negatively charged liposomes composed of phosphatidylserine (PS)/PC/Chol (1:1:1). However, the rate of uptake of PC/Chol liposomes at 37 degrees C is still 10-fold higher than that by fluid-phase pinocytosis. The uptake of liposomes, which is mediated by high-affinity binding to the cell surface binding sites and subsequent endocytosis, could be simulated and predicted by model calculations employing mass action kinetics. The number of binding sites, affinity constants of binding at 37 degrees C and 4 degrees C, on- and off-rate constants of binding, and endocytic rate constants for both types of liposomes were determined. The number of binding sites and the binding constants for PS/PC/Chol liposomes binding to J774 cells is severalfold to an order of magnitude higher than that for PC/Chol liposomes, but the rate constants at which they are endocytosed following binding to the cells are similar for both liposome types. The binding of liposomes, especially PS/PC/Chol, to J774 cells and monocytes is greatly enhanced by adherence of cells to plastic substratum and is also increased by maturation/differentiation in the case of monocytes. Our quantitative analysis indicates that the binding and endocytosis of liposomes, especially PS-containing liposomes, is mediated by binding sites that have strong affinity, comparable to or about an order of magnitude smaller than other known particle-cell interactions with specific receptors such as virus and lipoproteins binding to cells.

Animals↗

"Fast moving" and "slow moving" heparins, dermatan sulfate, and chondroitin sulfate: qualitative and quantitative analysis by agarose-gel electrophoresis.

Heparin from beef intestinal mucosa, dermatan sulfate from beef intestinal mucosa, and chondroitin sulfate from bovine trachea were extracted and purified, and their structures and physico-chemical properties were evaluated by different techniques (disaccharide patterns by specific enzymatic cleavage, relative molecular mass by high-performance size-exclusion chromatography, sulfate-to-carboxyl ratio by potentiometric determination). Heparin was fractionated into "slow moving" and "fast moving" fractions by selective precipitation as the barium salt at different temperatures. The "fast moving" and "slow moving" components of heparin, dermatan sulfate, and chondroitin sulfate were utilized to run calibration curves in agarose-gel electrophoresis. Mixtures containing different amounts of these glycosaminoglycans were made and separated by agarose-gel electrophoresis, and these were analyzed quantitatively. For analysis of relative amounts, the area of each individual component of mixtures, obtained by photodensitometric readings, was divided by the sum of the areas of all glycosaminoglycans and expressed as a percentage. For analysis of absolute amounts, the area under the curve for each component of mixtures was fitted to specific calibration curves, and the amount of each glycosaminoglycan was calculated in micrograms. The quantitative procedure performed by analysing absolute amounts was used to obtain an accurate quantitative evaluation of each component in mixtures of glycosaminoglycans utilized for pharmaceutical purposes. A sensitive method was developed for the evaluation of very small amounts (0.2% w/w) of possible glycosaminoglycans as contaminants in preparations of a single species of glycosaminoglycan. This technique requires specific enzymatic degradation by bacterial lyases, separation in agarose-gel electrophoresis, and quantitative analysis by photodensitometric analysis and specific calibration curves.

Animals↗

Insulin antibodies and hypoglycemia in diabetic patients. Can a quantitative analysis of antibody binding predict the risk of hypoglycemia?

We report a noninsulin-dependent diabetes mellitus (NIDDM) patient with spontaneous, severe hypoglycemic reactions and the presence of insulin antibodies. He had a remote antecedent history of beef-pork insulin therapy as well as exposure to hydralazine. Detailed insulin binding kinetic studies were performed in this patient as well as in six other insulin-treated diabetic patients with anti-insulin antibodies (three with and three without an obvious cause of hypoglycemia). Sera from the current patient and five of the six other diabetic patients (one NIDDM, four IDDM) revealed two types of binding sites: high-affinity with low capacity (Kd, 0.4-12.4 x 10(-9) mol/L; binding capacity, 0.6-659 mU/L) and low-affinity with high capacity (Kd, 0.3 to 35.7 x 10(-7) mol/L; binding capacity; 202-113,680 mU/L). One NIDDM patient had only high-affinity antibodies (Kd, 22.9 x 10(-9) mol/L; binding capacity of 78 mU/L). Type of diabetes mellitus, insulin antibody titers or their binding capacities, insulin levels (total, bound, or free), and bioavailable insulin were not related to hypoglycemic reactions. Two calculated values by the method described tended to discriminate patients with and without hypoglycemia. The calculated amount of low-affinity antibody bound insulin ranged from 69.4-2090 mU/L vs < 4-70.6 mU/L in patients with and without hypoglycemia, respectively. The best discrimination was afford by the percent saturation of low-affinity binding sites; values were clearly higher in the patients with hypoglycemia (2.5-34.4%) than in those without hypoglycemia (not detectable, 0.06, 0.15%). Consideration of the possible drug-associated insulin antibody formation in insulin-treated diabetics and the novel quantitative analysis of the antibody binding kinetics should prove helpful in evaluating patients with high insulin antibody titers and assessing the risk of hypoglycemia.

Adult↗

Quantitative analysis of Borrelia burgdorferi gene expression in naturally (tick) infected mouse strains.

Adaptation of Borrelia burgdorferi in the vector and vertebrate host is mediated by mechanisms that regulate differential expression of outer surface lipoproteins (Osps). In this study, real time PCR was applied to quantify tissue-specific expression of four linear plasmid (lp54)-encoded (ospA, zs7.a36, zs7.a66 zs7.a68) and one circular plasmid (cp26)-encoded (ospC) gene from B. burgdorferi sensu stricto, in a natural setting of tick-infected immunodeficient (C.B-17 SCID) and immunocompetent (BALB/c and AKR/OlaHsd) mice for up to 120 days post-infection (p.i.). Early during infection (day 30 p.i.) high numbers of spirochetes were found in the heart and joint, but not the ear and spleen tissues of disease-susceptible SCID mice. In disease-susceptible AKR mice spirochetes colonized the ear and joint tissues, but were undetectable in tissues of disease-resistant BALB/c mice. Later in infection (day 120 p.i.), spirochetes had expanded (approximately 1,000-fold) in all SCID tissues tested but were undetectable in AKR and BALB/c mice. Of the five genes analyzed, only zs7.a36 transcripts were detected in various tissues of all infected mouse strains, though at differing levels, whereas ospC transcripts were only found in tissue specimens of SCID mice. Furthermore, gene expression of ospC and zs7.a36 appears to be differentially regulated in distinct organs of individual mice. In contrast, transcripts for ospA, zs7.a66, and zs7.a68 were not detected in any of the mouse strains, independent of their immune status and/or the severity of their infection/inflammatory responses. Late during infection (day 120 p.i.), transcription of zs7.a36 and ospC was down-regulated in the tissues of SCID mice despite expansion of spirochetes. This type of quantitative analysis may be helpful to further disclose principles of pathogenesis of Lyme borreliosis and to design strategies for its therapeutic treatment.

Animals↗

Transneuronal degeneration in the midbrain central gray following chemical lesions in the ventromedial nucleus: a qualitative and quantitative analysis.

In the preceding experiments with electrolytic lesions of the ventromedial nucleus of the hypothalamus, we showed pre- and postsynaptic degeneration in the midbrain central gray of the rat. The postsynaptic degeneration seen may indicate a transneuronal effect of the ventromedial nucleus on the midbrain central gray. Electrolytic lesions, however, destroy afferent endings and fibers in passage, so that the postsynaptic degeneration seen in the midbrain central gray may be due to retrograde degeneration of midbrain central gray afferents to the ventromedial nucleus or due to degeneration of fibers in passage. In order to distinguish among these possibilities, chemical, i.e. kainic acid and N-methyl aspartate, lesions were made in the ventromedial nucleus and the ultrastructure of the midbrain central gray and cerebral cortex was examined at various intervals following the lesions. Both of these excitotoxins have been shown to destroy neurons, sparing afferent terminals and fibers in passage. Animals receiving kainic acid lesions in the right ventromedial nucleus were allowed to survive for one week, and animals receiving N-methyl aspartate lesions in the right ventromedial nucleus were permitted to survive for four, eight, and 20 days. Midbrain central gray tissue of unlesioned animals served as a control for both kainic acid and N-methyl aspartate lesions. In addition, other control animals received injections of the same amount of N-methyl aspartate in the right parietal cortex and were permitted to survive for four and eight days. For each of the above injection and survival conditions, the left cortex and subdivisions of the midbrain central gray were removed and processed for electron microscopy. Animals receiving ventromedial hypothalamic lesions with both kainic acid and N-methyl aspartate showed signs of pre- and postsynaptic degeneration. A quantitative analysis (General Linear Model Procedure) of degeneration was performed on the cortex and midbrain central gray of animals receiving N-methyl aspartate lesions in the ventromedial nucleus and cortex, and several parameters were measured. Animals receiving ventromedial hypothalamic lesions and surviving for eight and 20 days show significantly higher ratios of degenerating presynaptic elements to total presynaptic elements, degenerating postsynaptic elements to total postsynaptic elements, and degenerating total elements to total elements, in the midbrain central gray than in the cortex. Furthermore, the ratio of degenerating postsynaptic elements to total postsynaptic elements is larger than the other ratios.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Semi-quantitative analysis of telomerase activity in exfoliated human urothelial cells and bladder transitional cell carcinoma.

OBJECTIVE: To determine the utility of detecting telomerase activity in transitional cell bladder cancer, using the telomeric repeat amplification protocol (TRAP) assay, and thus provide a test for the detection and monitoring of bladder cancer, especially low-grade tumours. PATIENTS, SUBJECTS AND METHODS: Telomerase activity was assayed in bladder cancer tissues and the exfoliated urothelial cells from 29 patients with bladder cancer, 10 with benign disease, and in 10 healthy subjects using the TRAP assay. The levels were assessed semiquantitatively by calculating the ratio to an internal telomerase assay standard. RESULTS: Telomerase activity was very low in the exfoliated urothelial cells from all healthy subjects and patients with benign disease, with a mean (SD) ratio of 0.25 (0.03) and 0.33 (0.04), respectively. A threshold ratio of 0.4 was calculated as the mean +/- 2 SD of the telomerase activity level of the exfoliated urothelial cells from patients with benign disease. Using this threshold, telomerase activity was negative in exfoliated urothelial cells from all benign cases (100% specificity) and positive in all 26 bladder cancer tissue samples assayed. In tumour tissue, telomerase activity was not associated with tumour grade, size or stage. Telomerase activity in exfoliated urothelial cells from patients with bladder cancer was positive in 25 of 29 samples (86% sensitivity). The sensitivity of telomerase activity in exfoliated cells was seven of nine in G1, 10 of 12 in G2 and all eight G3 tumours; the corresponding sensitivity for voided urine cytology in G1, G2 and G3 tumours was two of nine, six of 12 and six of eight, respectively. CONCLUSION: These results indicate that telomerase activation occurs as an early step in carcinogenesis and the semi-quantitative analysis of telomerase activity in exfoliated urothelial cells could be a minimally invasive and useful method for detecting bladder cancer, even in low-grade tumours.

Aged↗

Head-direction cells recorded from the postsubiculum in freely moving rats. I. Description and quantitative analysis.

This paper is a study of the behavioral and spatial firing correlates of neurons in the rat postsubiculum. Recordings were made from postsubicular neurons as rats moved freely throughout a cylindrical chamber, where the major cue for orientation was a white card taped to the inside wall. An automatic video/computer system monitored cell discharge while simultaneously tracking the position of 2 colored light emitting diodes (LEDs) secured to the animal's head. The animal's location was calculated from the position of one of the LEDs and head direction in the horizontal plane calculated from the relative positions of the 2 LEDs. Approximately 26% of the cells were classified as head-direction cells because they discharged as a function of the animal's head direction in the horizontal plane, independent of the animal's behavior, location, or trunk position. For each head-direction cell, vectors drawn in the direction of maximal firing were parallel throughout the recording chamber and did not converge toward a single point. Plots of firing rate versus head direction showed that each firing-rate/head-direction function was adequately described by a triangular function. Each cell's maximum firing rate occurred at only one (the preferred) head direction; firing rates at head directions on either side of the preferred direction decreased linearly with angular deviation from the preferred direction. Results from 24 head-direction cells in 7 animals showed an equal distribution of preferred firing directions over a 360 degrees angle. The peak firing rate of head-direction cells varied from 5 to 115 spikes/sec (mean: 35). The range of head-direction angles over which discharge was elevated (directional firing range) was usually about 90 degrees, with little, if any, discharge at head directions outside this range. Quantitative analysis showed the location of the animal within the cylinder had minimal effect on directional cell firing. For each head-direction cell, the preferred direction, peak firing rate, and directional firing range remained stable for days. These results identify a new cell type that signals the animal's head direction in its environment.

Animals↗

Targeted profiling: quantitative analysis of 1H NMR metabolomics data.

Extracting meaningful information from complex spectroscopic data of metabolite mixtures is an area of active research in the emerging field of "metabolomics", which combines metabolism, spectroscopy, and multivariate statistical analysis (pattern recognition) methods. Chemometric analysis and comparison of 1H NMR1 spectra is commonly hampered by intersample peak position and line width variation due to matrix effects (pH, ionic strength, etc.). Here a novel method for mixture analysis is presented, defined as "targeted profiling". Individual NMR resonances of interest are mathematically modeled from pure compound spectra. This database is then interrogated to identify and quantify metabolites in complex spectra of mixtures, such as biofluids. The technique is validated against a traditional "spectral binning" analysis on the basis of sensitivity to water suppression (presaturation, NOESY-presaturation, WET, and CPMG), relaxation effects, and NMR spectral acquisition times (3, 4, 5, and 6 s/scan) using PCA pattern recognition analysis. In addition, a quantitative validation is performed against various metabolites at physiological concentrations (9 microM-8 mM). "Targeted profiling" is highly stable in PCA-based pattern recognition, insensitive to water suppression, relaxation times (within the ranges examined), and scaling factors; hence, direct comparison of data acquired under varying conditions is made possible. In particular, analysis of metabolites at low concentration and overlapping regions are well suited to this analysis. We discuss how targeted profiling can be applied for mixture analysis and examine the effect of various acquisition parameters on the accuracy of quantification.

Metabolism↗

[Focal liver lesions: characterization with quantitative analysis of T2 relaxation time in TSE sequence with double echo time].

PURPOSE: To evaluate the accuracy of calculation of T2 relaxation time with simplified algorithm in characterization of liver focal lesions with Turbo Spin Echo (TSE) T2-weighted sequences with double echo time (TE). MATERIALS AND METHODS: We carried out a retrospective analysis of 113 hyperintense hepatic focal lesions in T2-weighted sequences (52 metastases, 51 hemangiomas and 10 cysts). We have employed a single TSE T2-weighted sequence with effective TE of 83 and 165 msec and turbo factor 9. Signal intensity (IS) of each lesion was measured on different TE. Using these values, we calculated T2 relaxation time by the following simplified algorithm: T2(ms)=(TE2-TE1)/ (lnIS1-lnIS2). T2 relaxation time has been correlated with the final diagnosis, obtained by percutaneous biopsy, surgical resection or three months follow-up. RESULTS: The mean T2 relaxation time for metastases was 108,7 msec (+/-30), for hemangiomas 166,5 msec (+/- 29) and for cysts 312,4 msec (+/-19). The difference between the mean T2 relaxation time for metastases and hemangiomas was statistically significant (p<0.0001). Most metastases were best characterized between the threshold value of 130 and 140 msec. The values of specificity and sensitivity of the diagnosis of malignant hepatic lesion was respective 0,86 and 0,88. All false positive diagnoses were hemangiomas of less than one centimeter. CONCLUSIONS: Introducing double echo time TSE sequences in the protocol of liver study concurs to obtain high diagnostic accuracy in the characterization of the hepatic lesions. The quantitative analysis of T2 represents a useful additional instrument to the morphologic analysis in discrimination between hepatic lesions and can be carried out by TSE sequences with elevated reliability reducing the imaging time compared with conventional sequences. The evaluation of hepatic lesions of less than one centimeter should always include analysis of dynamic gadolinium-contrast images.

Adult↗

Quantitative analysis of EEG effects following experimental marginal magnesium and boron deprivation.

Magnesium (115 and 315 mg/d) and boron (0.23 and 3.23 mg/d) were fed in a double-blind Latin squares design to 13 healthy postmenopausal women (aged 50-78 years) living on a metabolic unit. An eight-channel electroencephalogram (EEG) was recorded during the last week of each of four 6-week dietary periods. Power and coherence measures were determined for each of four EEG frequency bands: delta (1-3 Hz), theta (4-7 Hz), alpha (8-12 Hz), and beta (13-18 Hz). Compared to high dietary magnesium, the low magnesium intake increased total power in the frontal regions and right temporal and parietal regions and resulted in frequency-specific increases in left occipital delta power, theta power in all but the left temporal region, alpha power in the right frontal and right temporal regions, and beta power in the frontal regions. The proportion of theta to total power in the parietal regions also increased with the low magnesium intake. While magnesium effects were observed primarily during eyes-closed conditions, effects of dietary boron on EEG power were found only during eyes-open conditions. Relative to high dietary boron, the low boron intake increased delta power in the left parietal and left occipital regions, increased the proportion of delta to total power in the frontal regions, and decreased relative right frontal theta, right frontal alpha, and left frontal beta power. Additional magnesium and boron effects were evident in the measures of EEG coherence. Thus relatively short periods of marginal magnesium and boron deprivation can affect brain function in healthy older women. The findings extend previous qualitative observations of increased CNS activity following severe magnesium deprivation and deficiency to cases of experimentally induced marginal magnesium deficiency, and verify CNS hyperexcitability by quantitative analysis of the EEG.

Adult↗

[Quantitative analysis of lateral force of floor reactions in normal and above-knee prosthetic gait].

Analysis of the lateral force of floor reactions has only been reported with reference to the vertical and fore- and-aft force because of large individual variations and great differences between the right and left limb of the same individual on the normal pattern of the force curve. In this study, the lateral force in normal and above-knee prosthetic gait was evaluated by calculating the total impulse during stance phase duration as well as by characteristic points of the force curve. The sensitivity of the electronic unit connected to the Kistler 's force plate was enhanced four times. The following results were obtained: In normal gait, the lateral stability during stance phase was shown by the lateral force which was proved to be a regulative element for rhythmical forward movement of the center of gravity of the body. The lateral stability in above-knee prosthetic gait was evaluated not only by the characteristic pattern of the force curve, but also by a decreased difference of the impulse of lateral force between the natural and artificial limb. The initial adduction angle of above-knee prostheses was determined by such a definition of the lateral force. Five degrees proved to be the most appropriate angle, as has been generally accepted. The lateral force of floor reaction can be definitely measured by a quantitative analysis of the total impulse during stance phase duration.

Adult↗

Noradrenergic innervation of vasopressin- and oxytocin-containing neurons in the hypothalamic paraventricular nucleus of the macaque monkey: quantitative analysis using double-label immunohistochemistry and confocal laser microscopy.

Previous reports on the rat and monkey hypothalamus have revealed a dense noradrenergic innervation within the hypothalamic paraventricular nucleus as assessed by dopamine-beta-hydroxylase immunohistochemistry. These single-label analyses were unable to delineate the cellular structures which receive this catecholaminergic innervation. Double-label preparations in the rat hypothalamic paraventricular nucleus have demonstrated synaptic interactions between noradrenergic varicosities and magnocellular neurons. However, the density and distribution of varicosities contacting chemically identified magnocellular neurons have not been assessed at the light or electron microscopic level. In this report, single-label immunohistochemistry was used to assess the morphology and distribution of vasopressin- and oxytocin-immunoreactive neurons within the macaque hypothalamic paraventricular nucleus. In addition, double-label immunohistochemistry was combined with confocal laser scanning microscopy to quantify the number of dopamine-beta-hydroxylase-immunoreactive varicosities in apposition to magnocellular neurons expressing vasopressin or oxytocin immunoreactivity. The morphology of chemically identified neurons was also compared to magnocellular neurons in the monkey hypothalamic paraventricular nucleus which were filled with Lucifer Yellow in order to assess the somatodendritic labeling of the immunohistochemical preparation. Qualitative assessment of immunohistochemically identified magnocellular cells indicated that vasopressin- and oxytocin-containing neurons are observed throughout the rostrocaudal extent of the monkey hypothalamic paraventricular nucleus, demarcating this structure from the surrounding anterior hypothalamus. The distribution of the two nonapeptides is complementary, with vasopressin-immunoreactive neurons having a greater somal volume and located in a more medial aspect of the mid and caudal hypothalamic paraventricular nucleus relative to oxytocin-immunoreactive perikarya. For the double-label preparations, a series of confocal optical sections was assessed through the total somal volume of vasopressin- and oxytocin-immunoreactive neurons along with the corresponding dopamine-beta-hydroxylase-immunoreactive varicosities in the same volume of tissue, generating a varicosity-to-neuron ratio which was further characterized morphologically to assess afferent input to the soma and proximal dendrites. Quantitative analysis revealed that vasopressin-immunoreactive neurons received approximately two thirds of their dopamine-beta-hydroxylase-immunoreactive varicosities in apposition to the proximal dendrites and one third in apposition to the somata. Furthermore, vasopressin-immunoreactive neurons received a greater innervation density than oxytocin-immunoreactive neurons, which did not have a differential distribution of varicosities on the proximal dendrites and somata. The distribution of dopamine-beta-hydroxylase-immunoreactive afferents on magnocellular neurons in the hypothalamic paraventricular nucleus may reflect a physiological role of this circuit in terms of preferential release of vasopressin from magnocellular neurons upon noradrenergic stimulation.

Animals↗