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Absolute quantitation in neurological PET: do we need it?

This article addresses the question posed in the title by examining the effects of parameters traditionally associated with improved absolute quantitation, on the analysis of 12 acquired immune deficiency syndrome dementia complex (ADC) patients compared to a normal control group. Results are discussed within the framework of the subprofile scaling model (SSM) for analyzing patterns of regional covariation. It is demonstrated that the ability to extract measures of group discrimination and disease progression are unaffected by (1) limited improvements in image resolution, (2) the use of transmission scan smoothing, (3) the application of a scatter deconvolution correction, and (4) converting region-of-interest measurements of counts per voxel to measurements of regional CMRglc. This "robustness" of the SSM approach is partly due to the extraction of disease-related subject weights, independent of any subject's global scaling effects. It is argued that other analysis techniques that initially reduce intersubject variation (e.g., using regional ratios or normalizing by global metabolic rates before applying traditional multivariate procedures) lack analytic features that may be important to identify multidimensional, disease-related image patterns. Based on the ADC patient data, it is concluded that measures of group discrimination and disease progression will not necessarily benefit from the organization of parameters traditionally associated with improved absolute quantitation.

AIDS Dementia Complex↗

Microsample preparation by dielectrophoresis: isolation of malaria.

An important enabling factor for realising integrated micro fluidic analysis instruments for medical diagnostics purposes is front-end sample preparation. Dielectrophoresis is a method that offers great potential for cell discrimination and isolation for sample processing, and here we have applied it to the problem of isolating malaria-infected cells from blood. During development of the malarial pathogen, Plasmodium falciparum, increases occur in the ionic permeability of the plasma membrane of infected erythrocytes. When challenged by suspension in a low conductivity medium, infected cells lose internal ions while uninfected cells retain them. The resultant dielectric differences between infected and uninfected cells were exploited by dielectrophoretic manipulation in spatially inhomogeneous, travelling electrical fields produced by two types of microelectrode arrays. Parasitised cells of ring form or later stage from cultures and clinical specimens were isolated by steric dielectric field-flow-fractionation, focused at the centre of a spiral electrode array, and identified and counted. The dielectrophoretic methods require only a few micro litres of blood, and should be applicable to the production of small, low-cost automated devices for assessing parasite concentrations with potential applicability to drug sensitivity studies and the diagnosis of malaria. By simple adjustment of the electrical field parameters, other cell subpopulations that characterise disease, such as residual cancer cells in blood, can be similarly isolated and analysed.

Animals↗

Force magnitude effects upon osteoprogenitor cells during premaxillary expansion in rats.

To study the effects of force magnitude on osteoprogenitor cell activity during premaxillary expansion, stainless steel helical springs were attached to the maxillary central incisors of 45 3-month-old male rats. The animals were randomly divided into force levels (0, 50, 100, 150, 200 gm) and were injected intraperitoneally with tritiated thymidine (1.0 uc/g wt.) 1 hour prior to sacrifice which occurred at 27, 40, and 60 hours. In order to examine cell activity within different regions of the suture, each premaxilla was divided into three geographic areas. Quantitative results were obtained by comparing the percent of labeled cells observed at different force levels, geographic areas, and observation times. The greatest number of labeled cells at each force level was found at 27 hours. Increased forces were correlated with increased numbers of labeled cells up to 100 gm, with decreased cell numbers at higher forces. The numbers of labeled cells at 200 gm were not significantly different from the controls. Histological observations of early bone formation at 60 hours supported the quantitative labeling results at 27 hours. The results also demonstrate a significant correlation between the geographic location of the labeled cells and force magnitude, with maximal cell stimulation occurring more superiorly in the suture as forces increased. The results suggest that early bone formation within the expanded suture can be maximized by varying force magnitude and distance from the point of force application.

Animals↗

Apheresis and the thrombotic thrombocytopenic purpura syndrome: current advances in diagnosis, pathophysiology, and management.

Endeavors to optimize the management of thrombotic thrombocytopenic purpura (TTP) syndrome and improve mortality and relapse rates are hindered by its poorly understood pathophysiology. Variability in the application of therapeutic plasma exchange (TPE), including replacement fluid strategies, desirable endpoints in the platelet count, serum lactate dehydrogenase concentration, and the use of a TPE taper, limit comparisons among published studies. The diversity of adjunctive therapies such as antiplatelet agents, steroids, and splenectomy further clouds comparisons. Recent progress in the diagnosis, pathophysiology, and management of TTP syndrome are summarized. The possible role of occult infection and newly emerging associations such as ticlopidine therapy are discussed. Advances in possible pathogenic mechanisms, the rationale for different replacement fluids including the recently licensed solvent-detergent treated plasma, and progress in the apheresis management of TTP syndrome are presented.

Blood Component Removal↗

Validity of a physical activity computer questionnaire in 12- to 18-year-old boys and girls.

The reliability and validity of a physical activity computer questionnaire of a usual week were studied in 33 adolescents between 12 and 18 years of age. Intraclass correlation coefficients and Kappa values were calculated to verify test-retest reliability. Validity was investigated by calculating Pearson correlation coefficients between the questionnaire and the Computer Science and Applications uniaxial accelerometer (CSA). Accelerometer data were obtained during seven successive days (sum and mean counts, estimated MET). Intraclass coefficients generally exceeded 0.70 and all Kappa values but one varied between 0.44 and 1.00. Transport variables (active transport from and to school, and during leisure time) showed no relationship with CSA. Sport participation during leisure time, sport participation summed with total transport, and the frequencies of moderate and hard activity were significantly correlated with CSA (r between 0.48 and 0.78). These data indicate that the physical activity computer variables provide reliable information. Moreover, sport participation (and summed with total transport) and the frequencies of moderate and hard activity provide valid data about adolescents' usual week physical activity, based on CSA comparison.

Adolescent↗

Single-target molecule detection with nonbleaching multicolor optical immunolabels.

We introduce and demonstrate the use of colloidal silver plasmon-resonant particles (PRPs) as optical reporters in typical biological assays. PRPs are ultrabright, nanosized optical scatterers, which scatter light elastically and can be prepared with a scattering peak at any color in the visible spectrum. PRPs are readily observed individually with a microscope configured for dark-field microscopy, with white-light illumination of typical power. Here we illustrate the use of PRPs, surface coated with standard ligands, as target-specific labels in an in situ hybridization and an immunocytology assay. We propose that PRPs can replace or complement established labels, such as those based on radioactivity, fluorescence, chemiluminescence, or enzymatic/colorimetric detection that are used routinely in biochemistry, cell biology, and medical diagnostic applications. Moreover, because PRP labels are nonbleaching and bright enough to be rapidly identified and counted, an ultrasensitive assay format based on single-target molecule detection is now practical. We also present the results of a model sandwich immunoassay for goat anti-biotin antibody, in which the number of PRP labels counted in an image constitutes the measured signal.

ATP-Binding Cassette Transporters↗

Single dose recombinant human erythropoietin reduces transforming growth factor beta in patients on chronic haemodialysis.

Short-term effects of recombinant human erythropoietin on serum levels of transforming growth factor beta-1, interleukin 1-alpha, interleukin 3, interferon gamma, and tumour necrosis factor alpha in patients with chronic renal failure on chronic haemodialysis were investigated. Recombinant human erythropoietin was applied subcutaneously in a dose of 75 IU/kg on 19 patients. Serum levels of transforming growth factor beta-1, interleukin 1-alpha, interleukin 3, interferon gamma, tumour necrosis factor alpha and erythropoietin, red blood cell parameters: red blood cell count, haemoglobin, haematocrit, and erythrocyte indices were determined before and after recombinant human erythropoietin single application. Transforming growth factor beta-1 serum levels were decreased after recombinant human erythropoietin (22.70 +/- 1.51 ng/ml versus 18.77 +/- 1.70 ng/ml (p < 0.01). None of the other investigated parameters was influenced significantly by recombinant human erythropoietin. Recombinant human erythropoietin in patients with chronic renal failure on chronic haemodialysis may influence anaemia not only through its stimulating effect on erythropoiesis, but also by direct oxygen-independent decrease of at least one of the negative regulators of erythropoiesis--the transforming growth factor beta.

Adult↗

Lipoteichoic acid from Staphylococcus aureus is a potent stimulus for neutrophil recruitment.

Lipoteichoic acid (LTA) is a major immunostimulatory principle of Gram-positive bacteria. Intranasal application of LTA from S. aureus to mice resulted in greatly increased neutrophil and macrophage counts in the bronchoalveolar lavage as well as increased levels of the chemokine KC. The potential of highly pure, bioactive LTA from S. aureus to induce neutrophil recruitment and activation was investigated further in the human system. Although neutrophils expressed the key known receptors, CD14, TLR2 and TLR6, LTA did not induce or prime neutrophils for oxidative burst, or release of chemokines, bactericidal permeability-increasing protein or myeloperoxidase. However, LTA induced a strong release of the chemoattractants LTB4, IL-8, C5a, MCP-1 and the colony-stimulating factor G-CSF in whole blood comparable to stimulation with the same concentration of LPS (S. abortus equi). Further, the cytokine and chemoattractant pattern induced by LTA correlated well with that induced by live S. aureus of the same strain. LTA does not appear to activate neutrophils directly, but is a strong stimulus for the recruitment of phagocytes to the site of infection.

Animals↗

Studies on composition and stability of a large membered bacterial consortium degrading phenol.

A ten member microbial consortium (AS) consisting of eight phenol-degrading and two non-phenol-degrading strains of bacteria was developed and maintained in a fed-batch reactor by feeding 500 mg l(-1) phenol for four years at 28 +/- 3 degrees C. The consortium could degrade 99% of 500 mg l(-1) phenol after 24 hours incubation with a biomass increase of 2.6 x 10(7) to 4 x 10(12) CFU ml(-1). Characterization of the members revealed that it consisted of 4 principal genera, Bacillus, Pseudomonas, Rhodococcus, Streptomyces and an unidentified bacterium. Phenol degradation by the mixed culture and Bacillus subtilis, an isolate from the consortium was compared using a range of phenol concentrations (400 to 700 mg l(-1)) and by mixing with either 160 mg l(-1) glucose or 50 mg l(-1) of 2,4-dichlorophenol in the medium. Simultaneous utilization of unrelated mixed substrates (glucose/2,4-dichlorophenol) by the consortium and Bacillus subtilis, indicated the diauxic growth pattern of the organisms. A unique characteristic of the members of the consortia was their ability to oxidize chloro aromatic compounds via meta pathway and methyl aromatic compounds via ortho cleavage pathway. The ability of a large membered microbial consortia to maintain its stability with respect to its composition and effectiveness in phenol degradation indicated its suitability for bioremediation applications.

Aerobiosis↗

Enumerative and binomial sequential sampling plans for soybean aphid (Homoptera: Aphididae) in soybean.

Since the discovery of the soybean aphid, Aphis glycines Matsumura, in midwestern U.S. soybean, Glycine max L., in 2000, the aphid has become a significant economic pest. Basic information about estimating population density within fields is unknown. Therefore, we developed two sampling plans to efficiently characterize A. glycines densities. Enumerative and binomial sequential plans were developed using 89 data sets collected from 10 commercial fields sampled during 2001-2003. Re-sampling software was used to validate the enumerative plan on whole plant counts, based on Taylor's power law parameters (a = 9.157 and b = 1.543). For research applications, the enumerative plan was modified to provide an actual precision level of 0.10 (SE/mean), which resulted in an average sample number of 310 individual plants. For integrated pest management (IPM) purposes, we developed an enumerative plan with an actual precision of 0.25, which resulted in an average sample number of 38 individual plants. For IPM applications, the binomial plan will likely be more practical. Binomial plans were developed using two tally thresholds at five action thresholds. Final analysis of the operating characteristic curve for each plan indicated that the tally threshold of > or = 40 aphids per plant, and an action threshold of 0.837 (84% of the plants infested) provided the most correct treat (4%) and no-treat (95%) decisions, with very low incorrect treat (0.5%) and no-treat (0.5%) decisions. A tally threshold of > or = 40 aphids per plant and action thresholds of 84% of plants infested is equivalent to a mean density of 250 aphids per plant, a recently recommended economic threshold. Using this threshold, the minimum required sample number for the binomial plan was 11 plants.

Agriculture↗

Wide dynamic range germanium detector for perturbation crystallography.

An IR detector based on a cooled germanium photodetector has been tested for applications in X-ray diffraction. The detector can be used simultaneously in photon-counting mode and current mode giving a dynamic range from < 1 to 10(9) photons s(-1). Since germanium is used as the photodetector, its efficiency at energies above 25 keV is much better than the silicon equivalents. The detector proved to be highly linear both in the low-flux region (< 10(5) photons s(-1)) where photon counting is used and in the high-flux region (> 10(5) photons s(-1)) where the detector is run in current mode. The response time of the detector is of the order of 1 mus, making it suitable for studies in perturbation crystallography, especially when coupled to a lock-in amplifier. As an example, the shift of a reflection of LiNbO(3) induced by an external electric field was determined with the germanium detector and lock-in amplifier.

Journal Article↗

Inhibition of Bacillus licheniformis LMG 19409 from ropy cider by enterocin AS-48.

AIMS: To determine the activity of enterocin AS-48 against ropy-forming Bacillus licheniformis from cider. METHODS AND RESULTS: Enterocin AS-48 was tested on B. licheniformis LMG 19409 from ropy cider in MRS-G broth, fresh-made apple juice and in two commercial apple ciders (A and B). Bacillus licheniformis was rapidly inactivated in MRS-G by 0.5 microg ml(-1)AS-48 and in fresh-made apple juice by 3 microg ml(-1). Concentration-dependent inactivation of this bacterium in two commercial apple ciders (A and B) stored at 4, 15 and 30 degrees C for 15 days was also demonstrated. Counts from heat-activated endospores in cider A plus AS-48 decreased very slowly. Application of combined treatments of heat (95 degrees C) and enterocin AS-48 reduced the time required to achieved complete inactivation of intact spores in cider A to 4 min for 6 microg ml(-1) and to 1 min for 12 microg ml(-1). D and z values also decreased as the bacteriocin concentration increased. CONCLUSION: Enterocin AS-48 can inhibit ropy-forming B. licheniformis in apple cider and increase the heat sensitivity of spores. SIGNIFICANCE AND IMPACT OF THE STUDY: Results from this study support the potential use of enterocin AS-48 to control B. licheniformis in apple cider.

Alcoholic Beverages↗

Feasibility of white-light time-resolved optical mammography.

We demonstrate the feasibility of white-light time-resolved optical mammography. The instrumentation is based on supercontinuum light generated in photonic crystal fiber and 32-channel parallel time-correlated single-photon-counting detection. Total measurement time is of the order of 10 min for typical clinical applications. Preliminary measurements performed on volunteers show the ability of the system to determine tissue constituent concentrations and structure over the entire breast area. Furthermore, measurements on a tissue-like sample demonstrate detection and characterization of inclusions.

Adult↗

Quantitative residual cortical activity measurement: appropriate test for diagnosis of renal artery stenosis?

OBJECTIVES: To evaluate the diagnostic validity of quantitative measurement of residual cortical activity (RCA) in renal artery stenosis (RAS). METHODS: In 45 patients with a high clinical likelihood of renovascular hypertension (RVH) and unimpaired renal function, dynamic imaging was performed after an intravenous bolus injection of 148 MBq (99m)Tc MAG3 for both baseline renoscintigraphy and captopril renography following oral application of 50 mg captopril. RCA was measured according to the Sfakianakis method: RCA = cortical counts at 20 min/counts at peak x100%. An increase in RCA of >/=5% from baseline was considered indicative of RAS. After renography, all patients underwent selective transfemoral angiography with the digital subtraction technique. A luminal reduction of >/=50% was considered as proof of RAS. RESULTS: The number of kidneys that had a change of >/=5% in RCA values was 12 (27.2%) in normal kidneys, 7 (58.3%) in the patients with bilateral RAS, 14 (82.3%) in the patients with unilateral RAS, and 21 (72.4%) in overall kidneys with RAS. The positive test ratio in pathologic groups was significantly higher than normal (p < 0.05). The sensitivity and specificity of the RCA test were 72.4 and 72.7%, respectively; the positive and negative predictive values were 63.6 and 80%, respectively. CONCLUSION: Quantitative measurements of RCA can be used as a diagnostic parameter of renal artery stenosis and may contribute of the diagnostic accuracy of visual interpretation and other renographic diagnostic criteria.

Angiography↗

Time-resolved Hadamard fluorescence imaging.

We present a new concept for fluorescence lifetime imaging (FLIM) based on time-resolved Hadamard imaging (HI). HI allows image acquisition by use of one single-point detector without requiring a moving scanning stage. Moreover, it reduces the influence of detector noise compared with raster scanning. By use of Monte Carlo simulations it could be confirmed that Hadamard transformation may decrease the error in lifetime estimation and in general in fluorescence parameter estimation when the signal-to-noise ratio is low and detector dark noise is high. This concept may find applications whenever the performance of FLIM or similar methods is limited by high dark-count rates and when the use of a single-point detector is preferable.

Algorithms↗

Comparative studies on the oxidative processes during phagocytosis measured by luminol-dependent chemiluminescence.

Four different methods, using luminol-enhanced chemiluminescence, are described for assessment of the oxidative processes associated with the phagocytosis of opsonized zymosan by granulocytes. The techniques described in the literature using whole blood as a substrate gave unreliable results due to quench effects of contaminating red blood cells, unpredictable serum interferences, or varying ratios of granulocytes and zymosan. In order to avoid these side effects, rapid and simple methods for the isolation of the granulocytes were applied, e.g. lysis of erythrocytes by ammonium chloride, or the isolation of granulocytes on a percoll gradient. The latter procedure proved to be reliable, simple and sufficiently quick for a routine application.

Ammonium Chloride↗

The concentrations of calcium buffering proteins in mammalian cochlear hair cells.

Calcium buffers are important for shaping and localizing cytoplasmic Ca2+ transients in neurons. We measured the concentrations of the four main calcium-buffering proteins (calbindin-D28k, calretinin, parvalbumin-alpha, and parvalbumin-beta) in rat cochlear hair cells in which Ca2+ signaling is a central element of fast transduction and synaptic transmission. The proteins were quantified by calibrating immunogold tissue counts against gels containing known amounts of each protein, and the method was verified by application to Purkinje cells in which independent estimates exist for some of the protein concentrations. The results showed that, in animals with fully developed hearing, inner hair cells had 110 of the proteinaceous calcium buffer of outer hair cells in which the cell body contained parvalbumin-beta (oncomodulin) and calbindin-D28k at levels equivalent to 5 mm calcium-binding sites. Both proteins were partially excluded from the hair bundles, which may permit fast unbuffered Ca2+ regulation of the mechanotransducer channels. The sum of the calcium buffer concentrations decreased in inner hair cells and increased in outer hair cells as the cells developed their adult properties during cochlear maturation. The results suggest that Ca2+ has distinct roles in the two types of hair cell, reflecting their different functions in auditory transduction. Ca2+ is used in inner hair cells primarily for fast phase-locked synaptic transmission, whereas Ca2+ may be involved in regulating the motor capability underlying cochlear amplification of the outer hair cell. The high concentration of calcium buffer in outer hair cells, similar only to skeletal muscle, may protect against deleterious consequences of Ca2+ loading after acoustic overstimulation.

Age Factors↗

Comparative study of methods for the quantification of biofilm on complete dentures.

This study compared the application and reliability of 4 methods for biofilm quantification (computerized, paper-weighing, point-counting, and planimetric) in complete dentures, verifying the correlation between them. The internal surfaces of 62 complete dentures were stained (5% erythrosine) and photographed. The slides were projected on paper, and the areas (total and biofilm-covered) were outlined with a pencil. These areas were measured with an equidistant point grid (point-counting method), a digital planimeter (planimetric method), and for the paper weighing method they were cut and weighed with a precision scale. For the computerized method, ImageTool software was used. In order to perform a validation test of the methods, all of them were applied to slide projections of geometric figures with known dimensions. The correlation tests showed high correlation values (r = 0.82 to 0.99) among the methods. The validation test (ANOVA) showed no statistically significant differences among the values obtained from the measurement of figures using all four quantitative methods and the real dimensions of these geometric figures. Quantitative methods were efficient and reliable for measuring quantity of biofilm in complete dentures, and may be useful in experimental studies on the efficacy of hygiene products. The computerized method was fast and easy to perform.

Aged↗