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Effect of experimental parameters on high b-value q-space MR images of excised rat spinal cord.

The influence of diffusion time (delta), gradient duration (delta), and TE on the appearance of high b-value q-space diffusion MR images of excised rat spinal cord (SC) was evaluated. The water signal decays in the white (WM) and gray matter (GM) were analyzed when the diffusion was measured perpendicular ( perpendicular) and parallel to the fibers of the SC, using three different approaches: single-component q-space analysis, the biexponential model, and the bi-Gaussian fit of the displacement distribution profile. Probability and displacement contrast and anisotropy indices were calculated for the WM and GM. It was found that WM/GM contrast increases as the diffusion time is increased when diffusion is measured perpendicular to the long axis of the SC. At a diffusion time of 50 ms, when diffusion was measured parallel to the fibers of the SC, the displacement was found to be higher for GM as compared to WM. For this direction the WM/GM contrast increased when diffusion time was increased, although here the changes were much less pronounced than for the perpendicular direction. The WM/GM displacement contrast nearly disappears for a diffusion time of 150 ms, when diffusion is measured parallel to the fibers of the SC. As expected, the anisotropy indices were found to be higher in WM than in GM, and increased with the increase in diffusion time. Both delta and TE affected the extracted parameters. It was found that long delta and long TE overemphasizes the apparent slow-diffusing water component of the SC, which is also the more restricted one. It is demonstrated that the single-component q-space analysis best describes diffusion in WM when diffusion is measured perpendicular to the fibers of the SC. In other cases, a more complete description is obtained by using two-component models.

Algorithms↗

SB 239063, a novel p38 inhibitor, attenuates early neuronal injury following ischemia.

The aim of the present study was to evaluate p38 MAPK activation following focal stroke and determine whether SB 239063, a novel second generation p38 inhibitor, would directly attenuate early neuronal injury. Following permanent middle cerebral artery occlusion (MCAO), brains were dissected into ischemic and non-ischemic cortices and Western blots were employed to measure p38 MAPK activation. Neurologic deficit and MR imaging were utilized at various time points following MCAO to monitor the development and resolution of brain injury. Following MCAO, there was an early (15 min) activation of p38 MAPK (2.3-fold) which remained elevated up to 1 h (1.8-fold) post injury compared to non-ischemic and sham operated tissue. Oral SB 239063 (5, 15, 30, 60 mg/kg) administered to each animal 1 h pre- and 6 h post MCAO provided significant (P<0.05) dose-related neuroprotection reducing infarct size by 42, 48, 29 and 14%, respectively. The most effective dose (15 mg/kg) was further evaluated in detail and SB 239063 significantly (P<0.05) reduced neurologic deficit and infarct size by at least 30% from 24 h through at least 1 week. Early (i.e. observed within 2 h) reductions in diffusion weighted imaging (DWI) intensity following treatment with SB 239063 correlated (r=0.74, P<0.01) to neuroprotection seen up to 7 days post stroke. Since increased protein levels for various pro-inflammatory cytokines cannot be detected prior to 2 h in this stroke model, the early improvements due to p38 inhibition, observed using DWI, demonstrate that p38 inhibition can be neuroprotective through direct effects on ischemic brain cells, in addition to effects on inflammation.

Animals↗

Latin American Cancer Research Coalition. Community primary care/academic partnership model for cancer control.

The Latin American Cancer Research Coalition (LACRC) was funded by NCI as a Special Populations Network to 1) provide training to clinic staff in cancer control and foster development of Latino faculty training, 2) conduct a needs assessment with the community clinics, 3) enhance the ability of the clinics to promote healthy lifestyles, 4) collaborate on research projects to improve use of early detection, and 5) explore partnerships to increase access to culturally competent cancer care. The LACRC developed a model for cancer control focused on community-based clinics as the focal point for in-reach and community outreach targeted to Latinos to reduce cancer disparities. This framework was designed to link the community to local hospitals and academic centers, build capacity, and promote diffusion of innovations directly into delivery systems. Eight research projects submitted by junior investigator/clinic teams have been funded by NCI. These research projects range from recruiting for clinical trials to prevention to survivorship. The LACRC has trained 6 cancer control coordinators from partner sites and educated 59 undergraduate minority student interns in aspects of cancer control research. Central to LACRC's success to date has been the creation and maintenance of an infrastructure of trusting relationships, especially those developed between clinician/investigators and individuals within the greater Latino community. Community clinics can be effective agents for cancer control among Latinos. Latinos are likely to participate in research conducted by culturally representative teams of researchers using culturally appropriate recruiting strategies. Cancer 2006. (c) 2006 American Cancer Society.

Central America↗

Review: movement of mRNA from transcription site to nuclear pores.

Pre-mRNA is transcribed primarily from genes located at the interface between chromatin domains and the interchromatin space. After partial or complete processing and complexing with nuclear proteins, the transcripts leave their site of synthesis and travel through the interchromatin space to the nuclear pores for export to the cytoplasm. It is unclear whether transcripts are tethered within the interchromatin space and move toward the nuclear pores using a metabolic energy-requiring, directed mechanism or, alternatively, move randomly by a diffusion-based process. We discuss here recent progress in understanding this step of gene expression, including our experiments tracking the movement of intranuclear poly(A) RNA in living cells. Our results and those of others are most consistent with a model in which newly synthesized mRNAs diffuse throughout the interchromatin space until they randomly encounter and are captured by the export machinery. Because the export machinery appears to preferentially bind transport-competent mRNAs (complexed with the correct complement of nuclear proteins), this diffusion-based model for intranuclear RNA movement potentially allows for a significant level of posttranscriptional control of gene expression.

Animals↗

Spatially varying steady state longitudinal magnetization in distant dipolar field-based sequences.

Sequences based on the distant dipolar field (DDF) have shown great promise for novel spectroscopy and imaging. Unless spatial variation in the longitudinal magnetization, Mz(s), is eliminated by relaxation, diffusion, or spoiling techniques by the end of a single repetition, unexpected results can be obtained due to spatial harmonics in the steady state MzSS(s) profile. This is true even in a homogeneous single-component sample. We have developed an analytical expression for the MzSS(s) profile that occurs in DDF sequences when smearing by diffusion is negligible in the TR period. The expression has been verified by directly imaging the MzSS(s) profile after establishing the steady state.

Magnetic Resonance Imaging↗

Line shapes in CP/MAS (13)C NMR spectra of cellulose I.

The CP/MAS (13)C NMR line shape of cellulose I has been qualitatively analyzed by direct simulations using the Ornstein-Uhlenbeck stochastic process and the Kubo model. Both approaches describe a anhydroglucose C4 carbon as a oscillator with fluctuating Larmor frequency. The NMR resonance frequency is written omega=omega +omega(t), where the fluctuating part with zero mean was modelled as a stationary Markov diffusion process. The simulation results both motivates the use of multiple line shapes when fitting CP/MAS (13)C NMR spectra recorded on cellulose I and gives some insights into why signals from crystalline cellulose I give rise to Lorentzian line shapes.

Carbohydrate Conformation↗

Lateral diffusion coefficients in membranes measured by resonance energy transfer and a new algorithm for diffusion in two dimensions.

We describe measurements of lateral diffusion in membranes using resonance energy transfer. The donor was a rhenium (Re) metal-ligand complex lipid, which displays a donor decay time near 3 micros. The long donor lifetime resulted in an ability to measure lateral diffusion coefficient below 10(-8) cm(2)/s. The donor decay data were analyzed using a new numerical algorithm for calculation of resonance energy transfer for donors and acceptors randomly distributed in two dimensions. An analytical solution to the diffusion equation in two dimensions is not known, so the equation was solved by the relaxation method in Laplace space. This algorithm allows the donor decay in the absence of energy transfer to be multiexponential. The simulations show that mutual lateral diffusion coefficients of the donor and acceptor on the order of 10(-8) cm(2)/s are readily recovered from the frequency-domain data with donor decay times on the microsecond timescale. Importantly, the lateral diffusion coefficients and acceptor concentrations can be recovered independently despite correlation between these parameters. This algorithm was tested and verified using the donor decays of a long lifetime rhenium lipid donor and a Texas red-lipid acceptor. Lateral diffusion coefficients ranged from 4.4 x 10(-9) cm(2)/s in 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DMPG) at 10 degrees C to 1.7 x 10(-7) cm(2)/s in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) at 35 degrees C. These results demonstrated the possibility of direct measurements of lateral diffusion coefficients using microsecond decay time luminophores.

Algorithms↗

The role of IFN-gamma in tumor transplantation immunity and inhibition of chemical carcinogenesis.

IFN-gamma contributes to the rejection of transplantable tumors and the inhibition of methylcholanthrene (MCA)-induced carcinogenesis by different mechanisms. In most tumor transplantation models, tumor rejection requires IFN-gamma receptor expression by host cells, but not by tumor cells. IFN-gamma produced by either CD4+ or CD8+ T cells acts on non-hematopoietic tumor stroma cells and, either directly or indirectly, induces angiostasis. This prevents rapid tumor burden and allows residual tumor cells to be eliminated. In some models, IFN-gamma also contributes to the destruction of existing tumor blood vessels. During MCA-induced tumorigenesis IFN-gamma is involved in the inhibition of MCA diffusion by encapsulation and reduction of DNA damage. This mechanism may primarily protect tissue from damage and simultaneously inhibit tumor development.

Animals↗

Dynamic NMR study of the mechanisms of double, triple, and quadruple proton and deuteron transfer in cyclic hydrogen bonded solids of pyrazole derivatives.

Using dynamic solid state (15)N CPMAS NMR spectroscopy (CP = cross polarization, MAS = magic angle spinning), the kinetics of the degenerate intermolecular double and quadruple proton and deuteron transfers in the cyclic dimer of (15)N labeled polycrystalline 3,5-diphenyl-4-bromopyrazole (DPBrP) and in the cyclic tetramer of (15)N labeled polycrystalline 3,5-diphenylpyrazole (DPP) have been studied in a wide temperature range at different deuterium fractions in the mobile proton sites. Rate constants were measured on a millisecond time scale by line shape analysis of the doubly (15)N labeled compounds, and by magnetization transfer experiments on a second timescale of the singly (15)N labeled compounds in order to minimize the effects of proton-driven (15)N spin diffusion. For DPBrP the multiple kinetic HH/HD/DD isotope effects could be directly obtained. By contrast, four rate constants k(1) to k(4) were obtained for DPP at different deuterium fractions. Whereas k(1) corresponds to the rate constant k(HHHH) of the HHHH isotopolog, an appropriate kinetic reaction model was needed for the kinetic assignment of the other rate constants. Using the model described by Limbach, H. H.; Klein, O.; Lopez Del Amo, J. M.; Elguero, J. Z. Phys. Chem. 2004,218, 17, a concerted quadruple proton-transfer mechanism as well as a stepwise consecutive single transfer mechanism could be excluded. By contrast, using the kinetic assignment k(2) approximately k(3) approximately k(HHHD) approximately k(HDHD) and k(3) approximately k(HDDD) approximately k(DDDD), the results could be explained in terms of a two-step process involving a zwitterionic intermediate. In this mechanism, each reaction step involves the concerted transfer of two hydrons, giving rise to primary kinetic HH/HD/DD isotope effects, whereas the nontransferred hydrons only contribute small secondary effects, which are not resolved experimentally. By contrast, the multiple kinetic isotope effects of the double proton transfer in DPBrP and of the triple proton proton transfer in cyclic pyrazole trimers studied previously indicate concerted transfer processes. Thus, between n = 3 and 4 a switch of the reaction mechanism takes place. This switch is rationalized in terms of hydrogen bond compression effects associated with the multiple proton transfers. The Arrhenius curves of all processes are nonlinear and indicate tunneling processes at low temperatures. In a preliminary analysis, they are modeled in terms of the Bell-Limbach tunneling model.

Journal Article↗

Spontaneous apoptosis in ovarian carcinomas: a positive association with p53 gene mutation is dependent on growth fraction.

Changes in cell survival contribute to tumour development, influence tumour biology and its response to chemotherapy. p53 gene alterations should negatively affect apoptosis by impaired p53-dependent apoptotic response. We looked for associations between spontaneous apoptosis, p53 gene mutation, p53 protein accumulation, growth fraction, bcl-2 expression and histological parameters in 64 ovarian, four tubal and three peritoneal carcinomas. Apoptotic cells were detected with the TUNEL method. p53 gene variants were detected by the single-strand conformation polymorphism and were sequenced directly. P53, Ki-67 and bcl-2 protein expressions were detected immunohistochemically. A weighed multiple logistic regression model was applied. Apoptotic index (AI) ranged 0.02-0.18 (mean 0.11); proliferation index (PI) ranged 3-90% (mean 54%). p53 gene mutations were present in 51, p53 protein accumulation in 46, and diffuse bcl-2 expression in 29 of 71 tumours. The AI was positively associated with the presence of p53 gene mutation (P = 0.011). However, the PI included into the analysis did positively influence the AI (P = 0.02) and diminished the association with p53 gene mutation (P = 0.082). The AI was negatively associated with good histological differentiation (P = 0.0006), the serous tumour type (P = 0.002), and diffuse bcl-2 expression (P = 0.025). Strong bcl-2 expression was associated with endometrioid tumour type (P = 0.002). FIGO stage and p53 protein accumulation were the only parameters that influenced overall survival time. Thus, our results suggest that histological tumour type and grade are major determinants of spontaneous apoptosis in ovarian carcinomas; p53 alterations do not adversely but rather positively affect spontaneous apoptosis by increasing growth fraction. This, in turn, suggests p53-independency of spontaneous apoptosis in ovarian carcinomas.

Adolescent↗

Temporal relation between the ADC and DC potential responses to transient focal ischemia in the rat: a Markov chain Monte Carlo simulation analysis.

Markov chain Monte Carlo simulation was used in a reanalysis of the longitudinal data obtained by Harris et al. (J Cereb Blood Flow Metab 20:28-36) in a study of the direct current (DC) potential and apparent diffusion coefficient (ADC) responses to focal ischemia. The main purpose was to provide a formal analysis of the temporal relationship between the ADC and DC responses, to explore the possible involvement of a common latent (driving) process. A Bayesian nonlinear hierarchical random coefficients model was adopted. DC and ADC transition parameter posterior probability distributions were generated using three parallel Markov chains created using the Metropolis algorithm. Particular attention was paid to the within-subject differences between the DC and ADC time course characteristics. The results show that the DC response is biphasic, whereas the ADC exhibits monophasic behavior, and that the two DC components are each distinguishable from the ADC response in their time dependencies. The DC and ADC changes are not, therefore, driven by a common latent process. This work demonstrates a general analytical approach to the multivariate, longitudinal data-processing problem that commonly arises in stroke and other biomedical research.

Animals↗

Polarization multiplexing and demultiplexing for appearance-based modeling.

Polarization has been used in numerous prior studies for separating diffuse and specular reflectance components, but in this work we show that it also can be used to separate surface reflectance contributions from individual light sources. Our approach is called polarization multiplexing and it has a significant impact in appearance modeling where the image as a function of illumination direction is needed. Multiple unknown light sources can illuminate the scene simultaneously, and the individual contributions to the overall surface reflectance are estimated. Polarization multiplexing relies on the relationship between the light source direction and the intensity modulation. Inverting this transformation enables the individual intensity contributions to be estimated. In addition to polarization multiplexing, we show that phase histograms from the intensity modulations can be used to estimate scene properties including the number of light sources.

Algorithms↗

Nuclear magnetic resonance microscopy of single neurons under hypotonic perturbation.

Nuclear magnetic resonance (NMR) characteristics of water in perfused single neurons undergoing a 20% hypotonic perturbation were examined quantitatively using NMR microscopy. The transverse relaxation times (T2) in the cytoplasm and nucleus increased by 24.0 +/- 8.5% (average +/- SE, n = 8) and 29.7 +/- 5.3% (n = 6), respectively, whereas the apparent diffusion coefficients (ADC) showed no significant change. These findings are consistent with the behaviors of a perfect osmometer and with accepted molecular relaxation and diffusion models and have significant impacts on current views of properties of cellular water. Furthermore, the results suggest that the increase of tissue intracellular-to-extracellular volume ratio during cell swelling is the predominant mechanism underlying the ADC reduction in acute brain ischemia. These data are the first direct quantitative measurements of the NMR characteristics of water in the cytoplasm and nucleus of single cells undergoing physiological perturbations and may lead to an improved diagnostic capability for NMR imaging in a variety of disease states.

Animals↗

A theoretical study of light fractionation and dose-rate effects in photodynamic therapy.

The efficacy of photodynamic therapy is dependent upon the optical dose rate or upon the fractionation schedule on the light. These effects are thought to be limited by the time required for oxygen diffusion from the capillaries, since this therapy can consume oxygen faster than it can be supplied to tissues distant from the blood vessels. Oxygen diffusion and consumption by metabolic and photochemical mechanisms have been modeled here to compare theoretical predictions with experimental results of varying light fractionations and delivered dose rates. The mathematics of the problem have been described in the literature, and the present study extends these calculations to allow a more direct and quantitative comparison with fractionation experiments, using both analytical and numerical arguments. The optimum fraction time was found to depend only on the intercapillary spacing and not on the intensity of irradiation or the concentration of photosensitizer. The calculations indicate that experimentally observed optimum fractionation times of 30 and 60 s correspond to a distance from capillary to cell of approximately 1 mm. These results suggest that the fractionated light irradiation experiments need careful interpretation, and some possible reasons for longer optimum fractionation times are discussed.

Dose-Response Relationship, Radiation↗

Comparison of two glare measurement methods through light scattering modeling.

Two methods used for evaluation of glare in patients with opacities of the ocular media are compared. One is a low contrast letter test and the other is a direct light scattering meter. Theoretical expressions for the measures obtained from the two different glare tests are derived in terms of the point spread function. Measurements on healthy test persons wearing diffusive glasses with known light scattering properties were in good agreement with the theory. Data from 26 cataractous eyes are presented and discussed in light of the theory.

Adult↗

Bioadhesive tablets for controlled transdermal delivery of drugs.

Transdermal bioadhesive tablets were formulated by the direct compression method and evaluated. Carboxyfluorescein (CF) was used as a model permeant. The compression parameters were kept consistent and the composition of the formulation was varied to alter the release rate of the drug from the formulations. The dosage form was evaluated for physicochemical, adherence, and in vitro diffusion parameters. The transdermal flux of the drug decreased with the increasing ratio of Ethyl cellulose (EC)/hydroxypropyl cellulose (HPC). The test formulations were subjected to pharmacokinetic studies in mice. The formulations were able to maintain a steady state plasma concentration up to 12 hours. They were found to be safe for transdermal use as interpreted from skin irritation studies carried out on rabbits.

Administration, Cutaneous↗

Revised estimates for continuous shoreline fumigation: a PDF approach.

A probability density function (PDF) fumigation model is presented here to study the dispersion of air pollutants emitted from a tall stack on the shoreline. This work considers dispersion of the pollutants in the stable layer and within the thermal internal boundary layer (TIBL) proceeds independently. The growth of TIBL is considered parabolic with distance inland. Turbulence is taken as homogeneous and stationary. Dispersion of particles (contaminant) in lateral and vertical directions is assumed independent of each other. This assumption allows us to consider the position of particles in both directions as independent random variables. The lateral dispersion distribution within the TIBL is considered as Gaussian and independent of height. A skewed bi-Gaussian vertical velocity PDF is used to account for the physics of dispersion due to different characteristics of updrafts and downdrafts within the TIBL. We have used Weil (J.C. Weil, A diagnosis of the asymmetry in top-down and bottom-up diffusion using a Lagrangian stochastic model, J. Atmos. Sci., 47 (1990) 501-515) solutions to find out the parameters of this PDF. Incorporating finite Lagrangian integral time scale for the vertical velocity component, it is observed that it reduces the vertical dispersion in the beginning and moves the point of maximum concentration further downwind. Due to little dispersion in the beginning, there is more plume to be dispersed causing higher concentrations at large distances. The model has considered Weil and Brower's (J.C. Weil, P.R. Brower, Estimating convective boundary layer parameters for diffusion applications, Maryland Power Plant Siting Program Rep. PPSP-MP-48, Department of Natural Resources, Annapolis, MD, 1985, 37 pp.) convective limit to analyze dispersion characteristics within TIBL. The revised model discussed here is evaluated with the data available from the Nanticoke field experiment on fumigation conducted in summer of 1978 in Ontario, Canada. The results of revised model are in good agreement with the observed data.

Air Pollutants↗

Serial magnetic resonance imaging in experimental primate stroke: validation of MRI for pre-clinical cerebroprotective trials.

Precise assessment of stroke outcome is critical for pre-clinical testing of cerebroprotective strategies. Differences in stroke volume measured by various magnetic resonance imaging (MRI) techniques are documented in humans, but not well described in experimental primate stroke. This study characterizes changes in stroke volume using serial MRI scans in a baboon model of reperfused cerebral ischemia. The location/area of hyperintensity on MRI corresponded with the TTC-stained infarct region. T2-weighted fast spin echo (T2W FSE), fluid attenuated inversion recovery (FLAIR), and diffusion weighted imaging (DWI) showed a decrease in infarct volume between 72 h and nine days post-ischemia (p = ns, p = 0.029, and p = 0.006). T2W FSE and FLAIR demonstrated an increase in infarct volume from 24 h to nine days post-ischemia, while DWI displayed a decrease over the same period. While early T2W FSE, FLAIR, and DWI all correlated with late infarct volume (p < 0.001), 72 h T2W FSE was the best direct measure (2.39% +/- 1.40% unity deviation). Serial MRI in a nonhuman primate model of focal cerebral ischemia recapitulates findings in clinical stroke. MRI at 72 h accurately predicts late infarct volume.

Animals↗