Biomedical subjects

David W Stanley

Publications and source records attributed to David W Stanley.

13 recordsLinked to original sources

PubMed2007

Anatomic compartments of the foot: a 3-Tesla magnetic resonance imaging study.

There is neither consensus on the number nor agreement on the location of the anatomic compartments of the foot. This project utilized high-resolution magnetic resonance imaging (MRI) to identify foot compartments. The purpose of this study was to devise a new system using 3-Tesla (3T) MRI that assessed the number and location of these compartments. Six feet from healthy volunteers were imaged. From these, 10 compartments were described: (1) medial, (2) calcaneal, (3) lateral, (4) central superficial, (5) central deep (adductor), (6-9) interossei, and (10) skin. The 3T MRI and foot/ankle coil allowed us to assess the number and location of foot compartments.

Compartment Syndromes↗

PubMed2005

3 Tesla MR imaging provides improved contrast in first-pass myocardial perfusion imaging over a range of gadolinium doses.

PURPOSE: To compare myocardial enhancement during first-pass myocardial perfusion imaging at 3.0 Tesla (T) and 1.5T. MATERIALS AND METHODS: First-pass myocardial perfusion imaging was performed on twelve normal subjects at 3T and 1.5T using an interleaved notched saturation recovery gradient echo pulse sequence. Subjects received either 0.10 mmol/kg for both scans (group 1), 0.075 mmol/kg for both scans (group 2), or 0.075 mmol/kg for the 3T scan and 0.10 mmol/kg for the 1.5T scan (group 3). RESULTS: Contrast enhancement was significantly greater at 3T than at 1.5T for the 12 subjects whether enhancement was normalized to baseline signal intensity (2.58 +/- 0.76 vs. 1.52 +/- 0.37, p < 0.0001) or to noise (57.6 +/- 19.7 vs. 14.7 +/- 7.8, p < 0001). For each of the three groups, contrast enhancement was significantly greater at 3T versus 1.5T (p < 0.0001, p < 0.001, p < 0.008 when normalized to baseline signal; p < 0.0001 for all groups when normalized to noise). CONCLUSION: 3T improves contrast in first-pass myocardial perfusion imaging at either 0.10 mmol/kg or 0.075 mmol/kg.

Adult↗

PubMed2004

Lipopolysaccharide evokes microaggregation reactions in hemocytes isolated from tobacco hornworms, Manduca sexta.

Insect cellular immune reactions to bacterial infection include nodule formation. Eicosanoids mediate several cellular actions in the nodulation process, including formation of hemocyte microaggregates, an early step. In previous work, we reported that isolated hemocytes produce and secrete eicosanoids that influence hemocyte behavior in response to bacterial challenge. We also reported that microaggregate formation in response to challenge was mediated by prostaglandins (PGs), but not by products of the lipoxygenase (LOX) pathways. In this paper we describe experiments designed to test the idea that exposing isolated hemocytes to lipopolysaccharide (LPS) evokes formation of hemocyte microaggregates and this cellular action is mediated by PGs. Results show that isolated hemocyte preparations challenged with LPS formed more hemocyte microaggregates than unchallenged preparations (6.9x10(3) microaggregates/ml hemolymph vs. 2.5x10(3) microaggregates/ml hemolymph). LPS challenge stimulated formation of hemocyte microaggregates in a dose dependent manner. Experimental groups pretreated with cyclooxygenase inhibitors produced fewer hemocyte microaggregates in response to LPS challenge than untreated control groups. The formation of hemocyte microaggregates was not influenced by LOX inhibitors. Furthermore, the influence of dexamethasone was reversed by supplementing the experimental groups with the eicosanoid precursor fatty acid molecule, arachidonic acid and PGH(2). Palmitic acid, which is not substrate for eicosanoid biosynthesis, did not reverse the effects of dexamethasone on the formation of microaggregates. The LOX product 5(S)hydroperoxyeicosa-6E,8Z,11Z,14Z-tetraenoic acid also did not reverse the effects of dexamethasone. These results are consistent with similar investigations performed with bacterial suspensions. We infer that isolated hemocyte preparations recognize and react to LPS by forming microaggregates and this reaction is mediated by PGs, but not products of the LOX pathway.

Animals↗

PubMed2004

Phospholipase A2 in salivary glands isolated from tobacco hornworms, Manduca sexta.

We describe a phospholipase A2 (PLA2) associated with the salivary glands of tobacco hornworms, Manduca sexta. This enzyme is able to hydrolyze arachidonic acid from the sn-2 position of PLs. Addition of the calcium chelator, EGTA, or calcium, to the Tris reaction buffer impaired the PLA2 activity, from which we infer the enzyme requires very low concentrations of calcium or perhaps other ions for optimal activity. PLA2 activity was sensitive to protein concentration (highest activity at 25 microg protein per microl), reaction time (optimal at 30 min), buffer pH (optimal at pH 8-10), and reaction temperature (optimal range 18-38 degrees C). The salivary gland PLA2 was sensitive to the site-specific inhibitor, oleyloxyethylphosphorylcholine and stable to freezing at -80 degrees C, but not -20 degrees C. The biological significance of this enzyme may relate to hydrolysis of fatty acid moieties from dietary PLs for absorption by midgut epithelia. This salivary gland enzyme may also be responsible for killing food-borne bacteria.

Animals↗

PubMed2004

An entomopathogenic bacterium, Xenorhabdus nematophila, inhibits hemocytic phospholipase A2 (PLA2) in tobacco hornworms Manduca sexta.

The entomopathogenic bacterium, Xenorhabdus nematophila, induces immunodepression in target insects and finally leads to lethal septicemia of the infected hosts. A hypothesis has been raised that the bacteria inhibit eicosanoid-biosynthesis pathway to interrupt immune signaling of the infected hosts. Here, we show direct evidence that X. nematophila inhibits the activity of phospholipase A2 (PLA2), the initial step in the eicosanoid-biosynthesis pathway. Inhibition of PLA2 was dependent on both incubation time with X. nematophila and the bacterial concentration in in vitro PLA2 preparations of Manduca sexta hemocytes. While living bacteria inhibited PLA2 activity, heat-killed X. nematophila rather increased PLA2 activity. X. nematophila secreted PLA2 inhibitor(s) which were detected in the organic, but not aqueous, extract of the bacterial culture medium. The PLA2 inhibitory activity of the organic extract was lost after heat treatment. These results clearly indicate that X. nematophila inhibits PLA2 activity, and thereby inhibits eicosanoid biosynthesis which leads to immunodepression of the infected hosts.

Analysis of Variance↗

PubMed2004

Myocardial viability: breath-hold 3D MR imaging of delayed hyperenhancement with variable sampling in time.

A method for visualizing myocardial infarction with a three-dimensional (3D) breath-hold gated acquisition was examined. By using variable sampling in time, whole heart coverage with a single volume acquisition was achieved in 24 heart beats. In a study of 35 patients, in whom 3D volume acquisition was compared with a two-dimensional (2D) acquisition, all regions of myocardial infarction were correctly identified at 3D examination. The mean imaging time for 12 section locations was 8.0 minutes +/- 3.0 with a 2D approach compared with 22 seconds +/- 4 with a 3D approach (P <.001). Advantages were also noted for infarct contrast-to-noise ratio: 60 +/- 37 for 3D versus 33 +/- 20 for 2D imaging (P <.001). No significant differences (P >.05) were noted at qualitative assessment of myocardial suppression, endocardial border visualization, respiratory and cardiac motion artifacts, or confidence of transmurality of the infarct.

Aged↗

PubMed2003

Eicosanoids in insect immunity: bacterial infection stimulates hemocytic phospholipase A2 activity in tobacco hornworms.

Intracellular phospholipase A(2) (PLA(2)) is responsible for releasing arachidonic acid from cellular phospholipids, and is thought to be the first step in eicosanoid biosynthesis. Intracellular PLA(2)s have been characterized in fat body and hemocytes from tobacco hornworms, Manduca sexta. Here we show that bacterial challenge stimulated increased PLA(2) activity in isolated hemocyte preparations, relative to control hemocyte preparations that were challenged with water. The increased activity was detected as early as 15 s post-challenge and lasted for at least 1 h. The increased activity depended on a minimum bacterial challenge dose, and was inhibited in reactions conducted in the presence of oleyoxyethylphosphorylcholine, a site-specific PLA(2) inhibitor. In independent experiments with serum prepared from whole hemolymph, we found no PLA(2) activity was secreted into serum during the first 24 h following bacterial infection. We infer that a hemocytic intracellular PLA(2) activity is increased immediately an infection is detected. The significance of this enzyme lies in its role in launching the biosynthesis of eicosanoids, which mediate cellular immune reactions to bacterial infection.

Animals↗

PubMed2003

The bacterium Xenorhabdus nematophilus depresses nodulation reactions to infection by inhibiting eicosanoid biosynthesis in tobacco hornworms, Manduca sexta.

The bacterium, Xenorhabdus nematophilus, is a virulent insect pathogen. We tested the hypothesis that this bacterium impairs insect cellular immune defense reactions by inhibiting biosynthesis of eicosanoids involved in mediating cellular defense reactions. Fifth instar tobacco hornworms, Manduca sexta, produced melanized nodules in reaction to challenge with living and heat-killed X. nematophilus. However, the nodulation reactions were much attenuated in insects challenged with living bacteria (approximately 20 nodules/larva for living bacteria vs. approximately 80 nodules/larva in insects challenged with heat-killed bacteria). The nodule-inhibiting action of living X. nematophilus was due to a factor that was present in the organic, but not aqueous, fraction of the bacterial cultural medium. The nodule-inhibiting factor in the organic fraction was labile to heat treatments. The immunodepressive influence of the factor in the organic fraction was reversed by treating challenged hornworms with arachidonic acid. The factor also depressed nodulation reactions to challenge with the plant pathogenic bacteria, Pseudomonas putida and Ralstonia solanacearum. These findings indicate that one or more factors from X. nematophilus depress nodulation reactions in tobacco hornworms by inhibiting eicosanoid biosynthesis.

Animals↗

PubMed2003

Prostaglandins, not lipoxygenase products, mediate insect microaggregation reactions to bacterial challenge in isolated hemocyte preparations.

Nodulation is the predominant cellular defense reaction to bacterial challenge in insects. Eicosanoids mediate several steps in the nodulation process, including formation of hemocyte microaggregations. Isolated hemocyte preparations synthesize and secrete eicosanoids, which mediate hemocytic immune reactions. Two major groups of eicosanoids are prostaglandins (products of cyclooxygenase pathways) and various products of lipoxygenase pathways. In this study, we test the hypothesis that prostaglandins, but not lipoxygenase products, mediate hemocyte microaggregation reactions in response to bacterial challenge. Our results indicate that isolated hemocyte preparations pretreated with the cyclooxygenase inhibitors indomethacin and naproxen yielded fewer microaggregates than untreated control groups (3.7 x 10(5) microaggregates/ml hemolymph vs. 11.0 x 10(5) microaggregates/ml hemolymph). These inhibitors influence hemocyte microaggregate formation in a dose-dependent manner in treatments ranging from 0 to 200 microM. The lipoxygenase inhibitors esculetin and caffeic acid did not impact the formation of microaggregates in this system. The influence of the phospholipase A(2) inhibitor dexamethasone was reversed by amending experimental (dexamethasone-treated) preparations with prostaglandin H(2), but not prostaglandin D(2), prostaglandin E(2), nor 5(S)-hydroperoxy-6E,8Z,11Z,14Z-eicosatetraenoic acid, a product of the lipoxygenase pathway. We infer that prostaglandins are the primary mediators of microaggregation reactions to bacterial challenge in insect hemocyte preparations.

Animals↗

PubMed2002

Prostaglandin biosynthesis by fat body from larvae of the beetle Zophobas atratus.

We describe prostaglandin (PG) biosynthesis by microsomal-enriched fractions of fat body prepared from larvae of the tenebrionid beetle, Zophobas atratus. PG biosynthesis was sensitive to incubation time, temperature, pH, substrate and protein concentration. Optimal PG biosynthesis conditions of those we examined included 2 mg of microsomal-enriched protein, incubated at 22 degrees C for 2 min at pH 6. These preparations yielded four major PGs: PGA(2), PGE(2), PGD(2) and PGF(2 alpha). PGA(2) and PGF(2 alpha) were the predominant eicosanoids produced under these conditions. Two non-steroidal anti-inflammatory drugs, indomethacin and naproxen, effectively inhibited PG biosynthesis in low concentrations. In vitro PG biosynthetic reaction conditions, using vertebrate or invertebrate enzyme sources, usually include a cocktail of reaction co-factors. The Z. atratus preparation similarly performs better in the presence of co-factors. Arch.

Animals↗

PubMed2002

Eicosanoids mediate Manduca sexta cellular response to the fungal pathogen Beauveria bassiana: a role for the lipoxygenase pathway.

Many studies have documented the involvement of eicosanoids in insect cellular immune responses to bacteria. The use of the fungal pathogen Beauveria bassiana as a nodulation elicitor, with inhibition of phospholipase A(2) by dexamethasone, extends the principle to fungi. This study also provides the first evidence of involvement of the lipoxygenase (LOX) pathway rather than the cyclooxygenase (COX) pathway in synthesis of the nodulation mediating eicosanoid(s). The LOX product, 5(S)-hydroperoxyeicosa-6E,8Z,11Z,14Z-tetraenoic acid (5-HPETE), substantially reversed nodulation inhibition caused by dexamethasone and the LOX inhibitors, caffeic acid and esculetin. The COX product, prostaglandin H(2) (PGH(2)), did not reverse the nodulation inhibition by dexamethasone or the COX inhibitor, ibuprofen. None of the inhibitors tested had a significant effect on the phagocytosis of B. bassiana blastospores in vitro. Hemocyte phenoloxidase activity was reduced by dexamethasone, esculetin, and the COX inhibitor, indomethacin. The rescue candidates 5-HPETE and PGH(2) did not reverse the inhibition.

Animals↗

PubMed2002

Dexamethasone inhibition of the cellular immune response of Drosophila melanogaster against a parasitoid.

Host larvae of Drosophila melanogaster injected with the eicosanoid biosynthesis inhibitor, dexamethasone, prior to parasitization by the wasp Leptopilina boulardi, exhibited significantly reduced rates of melanotic encapsulation in comparison with control and saline-injected larvae. The results of this investigation suggest that prostaglandins and other eicosanoids are involved as cell-signaling molecules in the hemocytic encapsulation reaction of D. melanogaster larvae.

Animals↗

PubMedDate not supplied

Parallel MR imaging: a user's guide.

Parallel imaging is a recently developed family of techniques that take advantage of the spatial information inherent in phased-array radiofrequency coils to reduce acquisition times in magnetic resonance imaging. In parallel imaging, the number of sampled k-space lines is reduced, often by a factor of two or greater, thereby significantly shortening the acquisition time. Parallel imaging techniques have only recently become commercially available, and the wide range of clinical applications is just beginning to be explored. The potential clinical applications primarily involve reduction in acquisition time, improved spatial resolution, or a combination of the two. Improvements in image quality can be achieved by reducing the echo train lengths of fast spin-echo and single-shot fast spin-echo sequences. Parallel imaging is particularly attractive for cardiac and vascular applications and will likely prove valuable as 3-T body and cardiovascular imaging becomes part of standard clinical practice. Limitations of parallel imaging include reduced signal-to-noise ratio and reconstruction artifacts. It is important to consider these limitations when deciding when to use these techniques.

Artifacts↗