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Experimental design of 31P MRS assessment of human forearm muscle function: restrictions imposed by functional anatomy.

The restrictions imposed by the functional anatomy of the finger flexor muscles on the experimental design of 31P MRS assessment of human forearm muscle function employing surface coil localization and voluntary exercise were investigated. It was found that 31P MRS metabolic data of finger flexor muscle should be correlated with mechanical data of combined flexion of only the ring and little fingers, rather than all four fingers as has been commonly the case in previously reported studies.

Adenosine Triphosphate↗

Experimentally-derived functional form for a population-averaged high-temporal-resolution arterial input function for dynamic contrast-enhanced MRI.

Rapid T(1)-weighted 3D spoiled gradient-echo (GRE) data sets were acquired in the abdomen of 23 cancer patients during a total of 113 separate visits to allow dynamic contrast-enhanced MRI (DCE-MRI) analysis of tumor microvasculature. The arterial input function (AIF) was measured in each patient at each visit using an automated AIF extraction method following a standardized bolus administration of gadodiamide. The AIFs for each patient were combined to obtain a mean AIF that is representative for any individual. The functional form of this general AIF may be useful for studies in which AIF measurements are not possible. Improvements in the reproducibility of DCE-MRI model parameters (K(trans), v(e), and v(p)) were observed when this new, high-temporal-resolution population AIF was used, indicating the potential for increased sensitivity to therapy-induced change.

Abdominal Neoplasms↗

Transfer function analysis of respiratory sinus arrhythmia: a measure of autonomic function in diabetic neuropathy.

Standard techniques measuring heart rate (HR) variability do not account for its dependence on the rate and depth of respiration or measure the time relationship between changes in lung volume and HR. We used transfer function analysis to determine the magnitude and time relationship of the HR response to a known change in lung volume in controls and diabetics. This technique demonstrated significant differences between controls and diabetics with varying degrees of autonomic dysfunction. Specifically, reduced supine vagal and increased supine sympathetic HR modulation was found with progression of the autonomic neuropathy. In response to postural change the normal diabetics displayed impaired sympathetic HR modulation. Transfer function analysis yields new insight into the sequence of changes that occur with diabetic autonomic neuropathy and provides an accurate, easily comprehensible measurement of respiratory induced HR variability.

Adult↗

Is it functional? Report on the first BSPR/EBI meeting on functional proteomics.

This report describes the first scientific meeting of the British Society for Proteome Research (BSPR), which was organised jointly with the European Bioinformatics Institute (EBI) and held in July 2004. The focus of the conference was functional proteomics with an emphasis on possible clinical application. The main subjects described here are: the need to simplify samples, the use of biological fluids versus tissue, consideration of biological and experimental variation and the creation of databases to achieve meaningful functional analysis.

Animals↗

Functional analysis of the slaty gene product (TRP2) as dopachrome tautomerase and the effect of a point mutation on its catalytic function.

Slaty (slt), an autosomal recessive mutation that arose in YZ57/Ch mice, results in the dilution of coat color and premature hair loss. Recently, a gene encoding a homologue of the melanogenic enzyme tyrosinase (termed tyrosinase related protein 2 or TRP2) was cloned and was subsequently mapped to the slaty locus on chromosome 14. TRP2 was shown to function in melanogenesis as DOPAchrome tautomerase by means of the catalytic activity of the immunopurified protein and in slaty mutant skin samples. In this study, we generated an expression vector of a murine TRP2 encoding cDNA and are able to confirm its activity as DOPAchrome tautomerase. We further demonstrate that the slaty mutation dramatically decreases the catalytic function of the protein.

Animals↗

Isolation and characterization of IS10 transposase separation of function mutants: identification of amino acid residues in transposase that are important for active site function and the stability of transposition intermediates.

IS10 transposase mediates end pairing, donor cleavage (or excision) and strand transfer reactions in IS10/Tn10 transposition. Details of how this multi-functional protein mediates these various steps have not been determined. We have used a genetic approach to help define structure-function relationships for IS10 transposase. IS10 transposase mutants that failed to form an excised transposon fragment in vivo and conferred a dominant-negative phenotype were isolated. Mutants with these properties were expected to be defective in catalysis and/or steps preceding donor cleavage. A total of seven mutants were identified, the corresponding proteins were purified and their activities assessed in vitro. Mutant EK292 was found to form wild-type levels of paired end complex, an early intermediate in the reaction, but to be defective in both donor cleavage and strand transfer. This phenotype suggests that E292 comprises part of a catalytic region. Mutants RH119, SF120 and RQ296 formed aberrant paired end complexes, indicating a defect in a step(s) preceding donor cleavage. Mutants WR98, IS101, and AV162 were found to promote donor cleavage in vitro but not strand transfer. For IS101 and AV162 the defect in strand transfer may be related to instability of excision complexes. Finally, the observation that IS101, RH119 and AV162 were greatly suppressed by Mn2+ suggests that these mutations have an effect on Mg2+ interactions with transposase.

Amino Acids↗

Fibroblast growth factor receptor function is required for the orderly projection of ganglion cell axons in the developing mammalian retina.

During the early stages of development various cell adhesion molecules (CAMs) and fibroblast growth factor receptors (FGFR) are expressed throughout the retinal neuroepithelium. The ability of retinal ganglion cells to project their axons to the optic fissure depends, in part, on cell-cell interactions mediated by cell adhesion molecules. In the present study we show that the ability of the firstborn rat retinal ganglion cells to extend axons in vitro can be stimulated by NCAM and L1, but not N-cadherin. Both CAM responses can be fully inhibited by antibodies that block neuronal fibroblast growth factor receptor function and by agents that block defined steps in the FGFR signal transduction cascade. When added to living E13.5 rat retinal whole-mount preparations the same agents induced errors in the orderly establishment of young axon patterns in the retinal periphery and caused axons in the retinal center to defasciculate. These results suggest that the activation of the fibroblast growth factor receptor signal cascade not only promotes survival and proliferation of various cell types but can also mediate intraretinal axon guidance.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Improved detection of event-related functional MRI signals using probability functions.

Selecting an optimal event distribution for experimental use in event-related fMRI studies can require the generation of large numbers of event sequences with characteristics hard to control. The use of known probability distributions offers the possibility to control event timing and constrain the search space for finding optimal event sequences. We investigated different probability distributions in terms of response estimation (estimation efficiency), detectability (detection power, parameter estimation efficiency, sensitivity to true positives), and false-positive activation. Numerous simulated event sequences were generated selecting interevent intervals (IEI) from the uniform, uniform permuted, Latin square, exponential, binomial, Poisson, chi(2), geometric, and bimodal probability distributions and fixed IEI. Event sequences from the bimodal distribution, like block designs, had the best performance for detection and the poorest for estimation, while high estimation and detectability occurred for the long-decay exponential distribution. The uniform distribution also yielded high estimation efficiency, but probability functions with a long tail toward higher IEI, such as the geometric and the chi(2) distributions, had superior detectability. The distributions with the best detection performance also had a relatively high incidence of false positives, in contrast to the ordered distributions (Latin square and uniform permuted). The predictions of improved sensitivities for distributions with long tails were confirmed with empirical data. Moreover, the Latin square design yielded detection of activated voxels similar to the chi(2) distribution. These results indicate that high detection and suitable behavioral designs have compatibility for application of functional MRI methods to experiments requiring complex designs.

Adult↗

TCR-independent T cell development mediated by gain-of-oncogene function or loss-of-tumor-suppressor gene function.

The mechanisms that govern differentiation of T cell precursors during intrathymic development bridge an interdisciplinary research field of immunology, oncology and developmental biology. Critical checkpoints controlling early thymic T cell development and homeostasis are set by the proper signaling function of the IL-7 receptor, c-Kit receptor, and the pre-T cell antigen receptor (pre-TCR). Given the intimate link between cell cycle control and differentiation in T cell development, proto-oncogenes and tumor suppressors participate as physiological effectors downstream of these receptors not only to influence the cell cycle but also to determine differentiation and survival. Gain- or loss-of-function mutations of these downstream effectors uncouples partially or completely T cell precursors from these checkpoints, providing a selective advantage and enabling aberrant development. These effectors can be identified by provirus tagging in normal mice and more readily by complementation tagging in mice with a predefined block in T cell differentiation.

Animals↗

Non-eicosanoid functions of essential fatty acids: regulation of adenosine-related functions in cultured neuroblastoma cells.

Studies have demonstrated that augmenting the omega 6 polyunsaturated-fatty-acid (PUFA) content of N1E-115 neuroblastoma cells by media supplementation with linoleic acid results in greater than or equal to 2-fold increases in basal levels of intracellular cyclic AMP (cAMP). Data suggested some involvement of increased production of adenosine from endogenous metabolites; however, increases in adenosine were not related to increased activity of 5'-nucleotidase or decreased uptake of extracellular adenosine. PUFA-dependent elevations in basal cAMP were evident within 1 min of exposure to a phosphodiesterase inhibitor; this phenomenon did not appear to be due to PUFA-dependent changes in Ca2+ uptake or to increases in sensitivity of adenylate cyclase to Ca2+. Forskolin-stimulated cAMP formation was 3-fold higher in PUFA-enriched cells than in control cells, which suggested a direct effect on the functioning of the catalytic unit. Linoleic acid supplementation resulted in a 2-fold increase in the maximum amounts of cAMP produced in response to the stable adenosine analogue, 5'-N'ethylcarboxy-amidoadenosine (NECA). The altered stimulatory response did not involve eicosanoid formation, but may have been related to an increase in the number of stimulatory adenosine receptors, as judged by binding of [3H]NECA. These studies indicate that membrane PUFA modulate adenosine-related functions in neuroblastoma cells, and suggest that a complex series of mechanisms is involved in this regulation.

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone↗

Restriction in vivo. V. Introduction of SOS functions in Escherichia coli by restricted T4 phage DNA, and alleviation of restriction by SOS functions.

Degradation products of restricted T4 DNA induced filamentation, mutagenesis, and to a lesser extent, synthesis of recA protein in wild type cells but not in recA, lexA or recBC mutants of Escherichia coli. We conclude that the structural damage to the DNA caused by restriction cleavage and exonuclease V degradation can induce SOS functions. Degradation of restricted nonglycosylated T4 DNA by exonuclease V delayed cell division and induced filament formation and mutagenesis in lexA+ but not in lexA- cells. Delay of cell division was also dependent upon recA and recBC functions. Such degradation of DNA also dramatically increased mutagenesis in tif- Sfi- cells at 42 degrees. The synthesis of recA protein continued in the restricting host after infection by the monglucosylated T4 phage, but enhanced synthesis is not induced to the extent seen in SOS induced tif- cells grown at 42 degrees. We also found that restriction of nonglycosylated T4 was alleviated in UV irradiated cells. The UV induced alleviation of rgl and rK restriction depended upon post irradiation protein synthesis and was not observed in recA, lexA or recBC mutants.

DNA Repair↗

Model identification and estimation of organ-function parameters using radioactive tracers and the impulse-response function.

Examination of the input-output events in functioning organs by the use of the impulse-response function (IRF) for a radioactive tracer is gaining more and more ground in nuclear medicine. This study summarizes the development of deconvolution analysis, laying special stress on the 'model-free' approach. System linearity and time invariance are discussed, and means of eliminating noise in IRFs originating from the input and organ-time-activity curves are outlined. Typical IRFs are illustrated by flow diagrams, time-domain curves, and their representation by Laplace transforms. The cases of nondiffusible and diffusible tracers as well as parenchymally extracted and transported substances are discussed. Methods for the derivation of models and for the calculation of physiologically important parameters from the IRFs are suggested.

Heart↗

Assessing levels of adaptive functioning: the Role Functioning Scale.

This paper reports data on reliability and validity of the Role Functioning Scale (RFS) a measure of level of functioning of adults in four domains. Psychometric properties were tested on an inner city sample of 112 psychiatrically disturbed and well, predominantly African-American, low-income mothers of young children. The RFS has good interitem, test-retest, and interrater reliability. The four scales and global RFS Index discriminated accurately between well and disturbed subjects. The Global RFS Index was significantly correlated with self-esteem and degree of disturbance. Individual scales demonstrated predicted relationships with quality of child-rearing and other independent behavioral indices. Results are discussed in terms of the unique information provided by the RFS and its potential contribution to treatment planning.

Activities of Daily Living↗

An ultrastructural comparison of neuromuscular relationships in blood vessels with functional and 'non-functional' neuromuscular transmission.

The neuromuscular relationships of the guinea-pig renal artery, which has previously been shown to be noradrenergically innervated but 'non-functional', i.e. not responsive to nerve stimulation in vitro, is compared with those of two arteries where functional neuromuscular transmission occurs: the carotid artery, where responses to nerve stimulation are slow and of medium amplitude, and the mesenteric artery, where the responses to nerve stimulation are relatively fast and of large amplitude. Substantial differences of ultrastructure were demonstrated. In the renal artery, there were no nerve varicosities closer than 400 nm to the smooth muscle layer, all varicosities had connective tissue interposed between them and the muscle wall, and there were significantly fewer varicosities (6 +/- 3.1 mm-1) compared with the carotid (33 +/- 8.3 mm-1; P less than 0.05) or mesenteric (105 +/- 15.8 mm-1; P less than 0.001) arteries. In the carotid artery, a small group of varicosities (17%), with little interposed connective tissue, had an average neuromuscular gap of 1.5 micron, whilst the remainder were separated by an average of 4.3 micron and by cellular and other connective tissue elements from the nearest smooth muscle cell. In the mesenteric artery, about half of the nerve varicosities were closer than 400 nm to the muscle layer and 25% of the varicosities had no connective tissue except basal lamina interposed in the neuromuscular space. Differences were also shown in the ultrastructural appearance of the nerves: analysis of the neuronal vesicles showed that the majority of varicosities were noradrenergic in all three vessels, with non-adrenergic varicosities forming 6% of the total in the carotid artery and 32% of the total in the mesenteric artery. Noradrenaline content was greater in the mesenteric artery (0.37 +/- 0.052 ng mm-2) compared with the carotid (0.17 +/- 0.017 ng mm-2; P less than 0.01) and renal (0.11 +/- 0.024 ng mm-2; P less than 0.01) arteries. These differences appear to correlate with the differences of neuromuscular activity in the three arteries.

Animals↗

The functional results after colectomy and ileorectal anastomosis for severe constipation (Arbuthnot Lane's disease) as related to rectal sensory function.

Rectoanal manovolumetry during graded isobaric rectal distension was carried out in 12 women with severe constipation classified as slow transit constipation (Arbuthnot Lane's disease). The resting anal sphincter pressure, the rectoanal inhibitory reflex and the rectal capacity were all normal. While the distension volumes required to elicit sensation of rectal filling and an urge to defaecate were within normal limits in all patients the distension pressures required to elicit such sensations fell outside the 95% limits of variation of control subjects in 4 patients. All patients were subsequently subjected to colectomy and ileorectal anastomosis. Patients with normal rectal sensory function had a satisfactory functional result after colectomy, whereas the four patients with blunted sensation did not improve. These findings suggest that rectoanal manovolumetry with determination of the distension pressures required to elicit rectal sensation is an important preoperative measure to be used in patients with severe constipation for selection of patients suitable for colectomy and ileorectal anastomosis.

Adult↗

From morphology to function: new neuro applications in functional magnetic resonance.

Functional Imaging of the brain is one of the most exciting recent developments in Magnetic Resonance Imaging (MRI). It refers to techniques which provide functional information about the human brain which goes beyond morphology. Examples are brain activation studies, perfusion and diffusion imaging. For example, the visualization of activated cortical brain regions during external stimulations or while performing specific tasks opens a completely new perspective for the use of MRI.

Artifacts↗

Functional magnetic resonance imaging of the primary motor cortex in humans: response to increased functional demands.

Functional magnetic resonance imaging (fMRI) studies have been performed on 20 right handed volunteers at 1.5 Tesla using echo planar imaging (EPI) protocol. Index finger tapping invoked localized activation in the primary motor area. Consistent and highly reproducible activation in the primary motor area was observed in six different sessions of a volunteer over a period of one month. Increased tapping rate resulted in increase in the blood oxygenation level dependent (BOLD) signal intensity as well as the volume/area of activation (pixels) in the contralateral primary motor area up to tapping rate of 120 taps/min (2 Hz), beyond which it saturates. Activation in supplementary motor area was also observed. The obtained results are correlated to increased functional demands.

Adult↗

A new liver functional study using Tc-99m DTPA-galactosyl human serum albumin: evaluation of the validity of several functional parameters.

Several parameters calculated with a new functional imaging agent for the liver, Tc-99m DTPA-galactosyl human serum albumin, were evaluated in 9 patients with liver cirrhosis, one with hepatocellular carcinoma, and five with both liver cirrhosis and hepatocellular carcinoma. LU3, which represents the cumulative uptake of the tracer from 3 to 4 minutes after injection, showed a strong correlation (r = 0.858, p = 0.0001) with LHL15, which represents the count ratio for the liver to sum for the liver and heart 15 minutes after injection of the tracer. It also showed a strong correlation (r = -0.896, p = 0.0001) with the indocyanine green retention rate (ICGR15). Regional ICGR15 is therefore calculable from the regional LU3. GSAR15, which represents the radioactivity of the tracer retained in the blood 15 minutes after injection, showed a strong correlation (r = 0.878, p = 0.0001) with HH15, which represents the count ratio for the heart 15 minutes after injection of the tracer divided by the count for the heart 3 minutes after injection. In conclusion, LU3 and GSAR15 are interesting and promising parameters for assessing liver function.

Adult↗