Biomedical subjects

David H Evans

Publications and source records attributed to David H Evans.

45 records · Page 3Linked to original sources

PubMed2003

Limits of uncertainty in measured values of embolus-to-blood ratio due to Doppler sample volume shape and location.

Measurements of the relative Doppler power scattered by an embolus and the surrounding blood are widely used to infer the composition of the embolus. For a given embolus, measured embolus-to-blood ratio (MEBR) is affected by the Doppler sample volume shape, the geometrical relationship between the sample volume and the vessel, and the embolus trajectory through the blood vessel. The likely magnitudes of such effects were quantified using a model that allowed calculation of theoretical values of MEBR as a function of the geometrical relationship between a blood vessel and a defined sample volume. Overall, the effects of embolus trajectory, likely insonation angles, and plausible vessel misalignments introduced uncertainties in MEBR values of approximately 10 to 12 dB for a given vessel size. In practice, the only operator-controlled factors are the position and orientation of the transcranial Doppler probe on the patients' heads. Probe positioning can significantly affect MEBR and suboptimal positioning may result in the reduced detection of emboli.

Embolism↗

PubMed2003

High-frequency genetic recombination and reactivation of orthopoxviruses from DNA fragments transfected into leporipoxvirus-infected cells.

Poxvirus DNA is not infectious because establishing an infection requires the activities of enzymes packaged in the virion. This barrier can be overcome by transfecting virus DNA into cells previously infected with another poxvirus, since the resident virus can provide the trans-acting systems needed to reactivate transfected DNA. In this study we show that cells infected with a leporipoxvirus, Shope fibroma virus (SFV), can reactivate vaccinia virus DNA. Similar heterologous packaging systems which used fowlpox-infected cells to reactivate vaccinia virus have been described, but SFV-infected cells promoted a far more efficient reaction that can produce virus titers exceeding 10(6) PFU/ micro g of transfected DNA. SFV-promoted reactions also exploit the hyperrecombinogenic systems previously characterized in SFV-infected cells, and these coupled recombination and reactivation reactions could be used to delete nonessential regions of the vaccinia virus genome and to reconstruct vaccinia virus from overlapping DNA fragments. SFV-catalyzed recombination reactions need only two 18- to 20-bp homologies to target PCR amplicons to restriction enzyme-cut vaccinia virus vectors, and this reaction feature was used to rapidly clone and express a gene encoding fluorescent green protein without the need for plaque purification or selectable markers. The ability of SFV-infected cells to reactivate fragments of vaccinia virus was ultimately limited by the number of recombinational exchanges required and one cannot reconstruct vaccinia virus from multiple PCR fragments spanning essential portions of the genome. These observations suggest that recombination is an integral part of poxvirus reactivation reactions and provide a useful new technique for altering the structure of poxvirus genomes.

Animals↗

PubMed2003

Postembolic spectral patterns of Doppler microembolic signals.

OBJECTIVES: This study was performed to examine the prevalence of postembolic spectral patterns in patients with prosthetic heart valves and evaluate their potential relation to the underlying embolic material. METHODS: The prevalence of the tail (TS) and lambda (LS) signs was examined in artifact signals and microembolic signals (MES) detected in 10 patients with prosthetic heart valves and 13 further patients with artificial heart valves during normobaria as well as during hyperbaric exposure (2.5 and 1.75 bar). RESULTS: A total of 1,721 MES were detected in the 10 patients; prevalence of TS and LS were 11 and 9.8%, respectively. Hyperbaric exposure in the second group of patients resulted in a significant increase of the prevalence of LS (13.7% at 2.5 bar and 9.2% at 1.75 bar, as compared to 7.7 and 8.2% during normobaria), while the prevalence of TS remained unchanged, ranging between 5 and 6.7%. Neither TS nor LS was observed following artifact signals. CONCLUSIONS: TS and LS showed a high specificity (100%) but low sensitivity (ranging between 7.7 and 11 and 5 and 9.8%, respectively) for the detection of MES. LS appear to be--at least partially--associated with gaseous embolic material.

Adult↗

PubMed2002

Cell signaling and ion transport across the fish gill epithelium.

A large array of circulating and local signaling agents modulate transport of ions across the gill epithelium of fishes by either affecting transport directly or by altering the size and distribution of transporting cells in the epithelium. In some cases, these transport effects are in addition to cardiovascular effects of the same agents, which may affect the perfusion pathways in the gill vasculature and, in turn, affect epithelial transport indirectly. Prolactin is generally considered to function in freshwater, because it is the only agent that allows survival of some hypophysectomized fish species in freshwater. It appears to function by either reducing branchial permeability, Na,K-activated ATPase activity, or reducing the density of chloride cells. Cortisol was initially considered to produce virtually opposite effects (e.g., stimulation of Na,K-activated ATPase and of chloride cell size and density), but more recent studies have found that this steroid stimulates ionic uptake in freshwater fishes, as well as the activity of H-ATPase, an enzyme thought to be central to ionic uptake. Thus, cortisol may function in both high and low salinities. Growth hormone and insulin-like growth factor appear to act synergistically to affect ion regulation in seawater fishes, stimulating both Na,K-activated ATPase and Na-K-2Cl co-transporter activity, and chloride cell size, independent of their effects on growth. Some of the effects of the GH-IGF axis may be via stimulation of the number of cortisol receptors. Thyroid hormones appear to affect seawater ion regulation indirectly, by stimulating the GH-IGF axis. Natriuretic peptides were initially thought to stimulate gill ionic extrusion, but recent studies have not corroborated this finding, so it appears that the major mode of action of these peptides may be reduction of salt loading by inhibition of oral ingestion and intestinal ionic uptake. Receptors for both arginine vasotocin and angiotensin have been described in the gill epithelium, but their respective roles and importance in fish ion regulation remains unknown. The gill epithelium may be affected by both circulating and local adrenergic agents, and a variety of studies have demonstrated that stimulation of alpha-adrenergic versus beta-adrenergic receptors produces inhibition or stimulation of active salt extrusion, respectively. Local effectors, such as prostaglandins, nitric oxide, and endothelin, may affect active salt extrusion as well as gill perfusion. Recent studies have suggested that the endothelin inhibition of salt extrusion is actually mediated by the release of both NO and prostaglandins. It is hoped that modern molecular techniques, combined with physiological measurements, will allow the dissection of the relative roles in ion transport across the fish gill epithelium of this surprisingly large array of putative signaling agents.

Adaptation, Physiological↗

PubMed2002

Leporipoxvirus Cu-Zn superoxide dismutase homologs inhibit cellular superoxide dismutase, but are not essential for virus replication or virulence.

Vertebrate poxviruses encode homologs of cellular cupro-zinc superoxide dismutases (Cu-Zn SOD). In this study we have examined the molecular genetic properties of two Cu-Zn SOD homologs encoded by the Shope fibroma virus (SFV) and myxoma virus. These Leporipoxvirus proteins should be catalytically inactive as judged by the point mutations which alter a key catalytic arginine and restructure the predicted Cu-binding domain. This prediction was confirmed using in situ gel assays and recombinant proteins produced both in bacteria and in mammalian cells. Western blot analysis showed that these proteins are produced in abundance late in infection and can, upon exposure to oxidizing conditions, form disulfide cross-linked dimers. They are also virion components and not essential for growth in culture or virulence. Leporipoxvirus Cu-Zn SOD homologs affected two phenotypes. First, deletion of the myxoma M131R gene caused the mutant virus to grow better ( approximately 10-fold) in culture than does the wild-type parent. Second, expression of either native or recombinant Leporipoxvirus proteins is accompanied by a decline in cellular Cu-Zn SOD activity. We concluded that these gene products can somehow modulate the activity of host Cu-Zn SODs, but what advantage is thus gained by the virus remains to be established.

Amino Acid Sequence↗

PubMed2002

Pendrin immunoreactivity in the gill epithelium of a euryhaline elasmobranch.

Pendrin is an anion exchanger in the cortical collecting duct of the mammalian nephron that appears to mediate apical Cl(-)/HCO3(-) exchange in bicarbonate-secreting intercalated cells. The goals of this study were to determine 1) if pendrin immunoreactivity was present in the gills of a euryhaline elasmobranch (Atlantic stingray, Dasyatis sabina), and 2) if branchial pendrin immunoreactivity was influenced by environmental salinity. Immunoblots detected pendrin immunoreactivity in Atlantic stingray gills; pendrin immunoreactivity was greatest in freshwater stingrays compared with freshwater stingrays acclimated to seawater (seawater acclimated) and marine stingrays. Using immunohistochemistry, pendrin-positive cells were detected on both gill lamellae and interlamellar regions of freshwater stingrays but were more restricted to interlamellar regions in seawater-acclimated and marine stingray gills. Pendrin immunolabeling in freshwater stingray gills was more apical, discrete, and intense compared with seawater-acclimated and marine stingrays. Regardless of salinity, pendrin immunoreactivity occurred on the apical region of cells rich with basolateral vacuolar-proton-ATPase, and not in Na(+)-K(+)-ATPase-rich cells. We suggest that a pendrin-like transporter may contribute to apical Cl(-)/HCO3(-) exchange in gills of Atlantic stingrays from both freshwater and marine environments.

Animals↗

PubMed2001

Advances in ultrasound.

Significant advances have been recently introduced into various fields of technology, taking advantage of the use of new piezoelectric materials and the large diffusion of broadband transducers. Various types of modulation may be applied to the pulse characteristics, using single pulse, multipulse or multiline techniques, and resulting in improved spatial resolution and better penetration. Non-linear imaging uses the harmonics component, which is generated by tissues or by contrast agents. Different modalities can be used to separate harmonics from fundamental bands from the received signal. New Doppler modes have been developed, whereas grey-scale flow imaging allows the simultaneous imaging of blood flow and tissues. Compounding techniques improve the contrast resolution of tissues and reduce artefacts. If 3D techniques are now currently available, real-time 4D imaging has been recently introduced. Elastographic imaging is still under evaluation, but promising clinical results have been shown. Recent release of the DICOM specification has made the full integration of ultrasound to the PACS systems easier. All these advances indicate that the contribution and potential of ultrasound in patient management is still growing.

Equipment Design↗

PubMedDate not supplied

Doppler ultrasound tracking instrument for monitoring blood flow velocity.

Doppler ultrasound (US) is potentially a valuable method for monitoring changes of blood flow velocity over a period of many minutes or even hours, but is seldom used in this way. One difficulty that may have contributed to this is the problem of maintaining a fixed geometry between the US beam and the blood vessel. A method of improving the success of monitoring might be to actively steer the US beam so as to maintain an adequate signal even when small displacements of the transducer occur. We have designed and built a prototype system for this purpose. The system comprises a continuous-wave phased-array transducer controlled by a purpose-built Doppler unit. The system constantly evaluates the quality of the returning Doppler signal in terms of total power and signal-to-noise ratio (SNR) (evaluated by assessing the quality of derived envelope signals), and steers the ultrasonic beam in a manner so as to improve the signal, should this be necessary. The system was tested in vitro, where the automatic tracking of the Doppler signal doubled the effective beam width of the transducer. Further developments that increase sensitivity and steering range should result in US Doppler systems that are better suited to long-term monitoring.

Blood Flow Velocity↗

PubMedDate not supplied

Developmental origin of shark electrosensory organs.

Vertebrates have evolved electrosensory receptors that detect electrical stimuli on the surface of the skin and transmit them somatotopically to the brain. In chondrichthyans, the electrosensory system is composed of a cephalic network of ampullary organs, known as the ampullae of Lorenzini, that can detect extremely weak electric fields during hunting and navigation. Each ampullary organ consists of a gel-filled epidermal pit containing sensory hair cells, and synaptic connections with primary afferent neurons at the base of the pit that facilitate detection of voltage gradients over large regions of the body. The developmental origin of electroreceptors and the mechanisms that determine their spatial arrangement in the vertebrate head are not well understood. We have analyzed electroreceptor development in the lesser spotted catshark (Scyliorhinus canicula) and show that Sox8 and HNK1, two markers of the neural crest lineage, selectively mark sensory cells in ampullary organs. This represents the first evidence that the neural crest gives rise to electrosensory cells. We also show that pathfinding by cephalic mechanosensory and electrosensory axons follows the expression pattern of EphA4, a well-known guidance cue for axons and neural crest cells in osteichthyans. Expression of EphrinB2, which encodes a ligand for EphA4, marks the positions at which ampullary placodes are initiated in the epidermis, and EphA4 is expressed in surrounding mesenchyme. These results suggest that Eph-Ephrin signaling may establish an early molecular map for neural crest migration, axon guidance and placodal morphogenesis during development of the shark electrosensory system.

Animals↗