Biomedical subjects

W D Smith

Publications and source records attributed to W D Smith.

At least 37 records · Page 2Linked to original sources

PubMed2001

Approaches to vaccines for Psoroptes ovis (sheep scab).

A brief outline of the history of sheep scab in the UK is presented together with the current chemical methods used for its control and problems associated with these. Possible alternative approaches to control are discussed, as are selected aspects of the physiology of Psoroptes ovis and the pathogenesis of the scab lesion from the perspective of control through immunisation. Evidence is provided that immunity to the disease can indeed be acquired, both naturally after a previous infection and following inoculation of mite proteins in adjuvant. These results support the view that control by vaccination may be possible, although little is known to date about the antigens involved or the mechanism of protection.

Animal Feed↗

PubMed2001

Dynamic ventilatory response to acute isocapnic hypoxia in septuagenarians.

This study compared the ventilatory response to 20 min of acute isocapnic hypoxia (end-tidal P(O(2)), 50 mmHg) using the technique of dynamic end-tidal forcing in young (Y) and old (O) men. Two groups of non-smoking male subjects (mean +/- s.d. age: Y, 29.8 +/- 6.9 years; O, 73.4 +/- 2.8 years) with similar body size, normal age-predicted spirometry, and normal moderate levels of physical activity were studied. Compared with baseline ventilation in euoxia (10.79 +/- 1.99 and 11.88 +/- 0.91 l min-1) both groups responded to the abrupt onset of isocapnic hypoxia with peak ventilatory responses of 22.58 +/- 2.60 and 24.56 +/- 2.54 l min-1 for Y and O, respectively (not significant, n.s.). Both groups demonstrated a significant increment in neuromuscular drive (i.e. tidal volume (V(T))/inspiratory time (T(I)); 0.46 +/- 0.06 to 0.91 +/- 0.15 and 0.48 +/- 0.06 to 0.91 +/- 0.12 l s-1 for Y and O, respectively) with a small (but also significant) change in central timing (T(I)/total ventilation time (T(tot)); 0.38 +/- 0.02 to 0.41 +/- 0.02 and 0.42 +/- 0.02 to 0.45 +/- 0.02 for Y and O, respectively). Oxygen sensitivity was assessed using Weil's equation, and gave a hyperbolic factor (A) of 282 +/- 75 and 317 +/- 72, and using the linear equation: change in expiratory minute volume (DeltaV.(E))/change in arterial O(2) saturation (DeltaS(a,O(2))) which gave -1.17 +/- 0.57 and -1.17 +/- 0.42 l min-1 %-1 (n.s.) for Y and O, respectively. After 20 min of sustained isocapnic hypoxia, ventilation declined to 14.29 +/- 1.92 and 16.85 +/- 2.34 l min-1 for Y and O, respectively (n.s.). The acute response to hypoxia was characterised by similar time constants (16.0 +/- 5.4 and 18.5 +/- 6.7 s) and time delays (4.8 +/- 2.1 and 4.6 +/- 1.9 s) for Y and O, respectively. Thus, the dynamic ventilatory response to acute isocapnic hypoxia is maintained into the eighth decade in a group of habitually active elderly men. Experimental Physiology (2001) 86.1, 117-126.

Acute Disease↗

PubMed2000

Attempts to control haemonchosis in grazing ewes by vaccination with gut membrane proteins of the parasite.

A vaccination trial was conducted to evaluate the potential benefit of Haemonchus contortus gut membrane proteins as vaccine antigens under field conditions in Louisiana. The trial was conducted in the summer of 1996 in a flock of ewes grazing pasture naturally infected with H. contortus. Ewes were randomly assigned to three treatment groups (vaccine, adjuvant only, and saline) and fecal egg counts (FEC, measured as eggs per gram of feces), packed cell volumes (PCV), and antibody levels were monitored fortnightly for 12 weeks. It was shown by FEC that there were large individual variations in susceptibility to H. contortus in both vaccinated and non-vaccinated sheep, a finding which could have masked differences between treatments when analyzed by conventional statistical methods. Based on their egg counts before the period when the vaccination could have had an effect, all ewes were categorized as 'susceptible' or 'relatively resistant'. The significance of differences between FEC, PCV and antibody responses of vaccinated and control sheep were tested separately for the 'susceptible' and 'relatively resistant' category. The 'susceptible' vaccinates shed 65% fewer worm eggs during the period when the vaccine could have had an effect, but the difference was only significant on Week 6 post-vaccination. In these experiments, it was difficult to completely exclude the confounding effect of having 'relatively resistant' sheep in the control group. More studies are needed to further evaluate H11 and H-gal-GP antigens under field conditions.

Animals↗

PubMed2000

Localisation and characterisation of ovine immunglobulin within the sheep scab mite, Psoroptes ovis.

Ovine IgG was detected in homogenates of repeatedly washed Psoroptes ovis. Some of the immunoglobulin in the homogenates was fragmented although the host IgG present in mite washings was largely intact. The host immunoglobulin was immuno-localised to the surface or cytoplasm of the gut cells of feeding stages of freshly harvested P. ovis examined by cryosectioning. A similar distribution of rabbit IgG was detected in P. cuniculi. The IgG demonstrated in the mite gut represented partially digested as well as intact immunoglobulin. The presence of intact host immunoglobulin suggests that P. ovis may be susceptible to vaccination by the gut antigen approach, a method used successfully for blood-feeding ectoparasites like Boophilus microplus.

Animals↗

PubMed2000

Relative protective properties of three membrane glycoprotein fractions from Haemonchus contortus.

Jacalin lectin was used as a ligand to isolate a fraction containing two distinct protective antigens from detergent extracts of membranes from Haemonchus contortus. The first antigen was identified as a complex which appeared very similar to Haemonchus galactose-containing glycoprotein (H-gal-GP), which is a previously described protective protease complex, except that it was substantially depleted of one of the main H-gal-GP components, a 230 kDa metallopeptidase-containing band. The new complex was termed Haemonchus sialylated galactose-containing glycoprotein (H-sialgal-GP), because it bound to jacalin but not to peanut lectin and only jacalin will bind the sialylated form of galactosyl (beta-1, 3) N-acetylgalactosamine. Two protection trials with sheep showed that H-sialgal-GP and H-gal-GP were equally efficacious, reducing numbers of Haemonchus eggs by between 86% and 93% and worms by between 52% and 75%, respectively. The second jacalin-binding protective antigen fraction was separated from H-sialgal-GP by ion exchange and gel filtration chromatography. It was greatly enriched for two proteins termed p46 and p52 according to their apparent molecular weights. Immunization of sheep with these proteins gave protection values of 78% for eggs and 33% for worms, which are significantly lower than those obtained with either H-gal-GP or H-sialgal-GP. N-terminal amino acid sequence data from p46 and p52 showed that both proteins were closely related to a previously described 45 kDa Haemonchus membrane protein, which had conferred protection against Haemonchus in guinea-pigs.

Amino Acid Sequence↗

PubMed2000

Evaluation of immunization with gut membrane glycoproteins of Ostertagia ostertagi against homologous challenge in calves and against Haemonchus contortus in sheep.

Peanut and ConA lectins were used as ligands to isolate glycoproteins from detergent extracts of adult Ostertagia ostertagi membranes. As judged by their profiles following SDS-PAGE, these fractions closely resembled the equivalents from Haemonchus contortus which are derived from the nematode intestinal cell microvillar membranes and which are highly protective when used as antigens. Groups of calves were immunized with the peanut and ConA binding fractions of Ostertagia, either as separate or pooled antigens mixed with QuilA as adjuvant. All calves, including controls immunized with adjuvant only, were challenged with a single dose of infective Ostertagia larvae and faecal egg counts were monitored for 5 weeks. In two experiments where the antigen fractions were pooled, moderate (30-50%), but statistically significant reductions in egg output were observed, but the number of worms was not diminished. No significant protection was observed in a third trial where groups of calves were immunized with peanut or ConA binding proteins given separately. Two further trials were conducted in sheep immunized with the same Ostertagia fractions but challenged with Haemonchus. Irrespective of whether they were administered separately or together, the Ostertagia antigens cross protected efficiently against Haemonchus reducing egg counts by between 81% and 97% and worm numbers by between 57% and 84%.

Adjuvants, Immunologic↗

PubMed2000

A virtual instrument ergonomics workstation for measuring the mental workload of performing video-endoscopic surgery.

The visual and physical interface imposed on the surgeon by video-endoscopic surgery (VES) increases the surgeon's mental workload. Ergonomic studies are needed to develop ways to reduce this workload. We used virtual instrumentation to devise a portable ergonomic workstation to compare the surgeon's mental workloads during simulated open surgery and VES. The system measures palmar tonic skin conductance level (SCL) and electrooculogram (EOG) and frontalis electrical activity to monitor mental stress and concentration levels. We used the system at a national surgery conference on volunteer subjects during a rest period and as they performed simulated surgery, consisting of typing knots using open and VES techniques. The subjects were asked to self-rate their levels of mental concentration and stress during these activities and reported that both progressively increased from rest to the open surgery task to the VES task. The subjects tied fewer knots during the VES than the open task, consistent with the increased demands of the VES task. The SCL progressively increased from rest to the open task to the VES task, correlating with the subjects' reported increase in mental stress level. Eye blinks and low frequency EOG activity decreased from rest to the open task, consistent with the subjects' reported increase in mental concentration level. From the open to the VES task, eye blinks and EOG activity increased, as expected given the greater demands of the VES task. High frequency frontalis activity merits further study as another indicator of the subjects' levels of mental concentration and stress.

Computer Simulation↗

PubMed1999

Propagating distributions up directed acyclic graphs.

In a previous article, we considered game trees as graphical models. Adopting an evaluation function that returned a probability distribution over values likely to be taken at a given position, we described how to build a model of uncertainty and use it for utility-directed growth of the search tree and for deciding on a move after search was completed. In some games, such as chess and Othello, the same position can occur more than once, collapsing the game tree to a directed acyclic graph (DAG). This induces correlations among the distributions at sibling nodes. This article discusses some issues that arise in extending our algorithms to a DAG. We give a simply described algorithm for correctly propagating distributions up a game DAG, taking account of dependencies induced by the DAG structure. This algorithm is exponential time in the worst case. We prove that it is #P complete to propagate distributions up a game DAG correctly. We suggest how our exact propagation algorithm can yield a fast but inexact heuristic.

Algorithms↗

PubMed1999

Ergonomic problems associated with laparoscopic surgery.

BACKGROUND: The Society of American Gastrointestinal Endoscopic Surgeons (SAGES) Task Force on Ergonomics conducted a subjective and objective assessment of ergonomic problems associated with laparoscopic instrument use. The goal was to assess the prevalence, causes, and consequences of operational difficulties associated with the use of laparoscopic instruments. METHODS: A questionnaire was distributed asking respondents to rate the frequency with which they experienced pain, stiffness, or numbness in several body areas after laparoscopic operations. An ergonomics station was assembled to quantify forearm and thumb muscle workload. Processed electromyogram (EMG) signals were acquired from 27 volunteer surgeon subjects while they completed simulated surgical tasks using a hemostat and an Ethicon laparoscopic grasper, with the aid of an endoscopic trainer and video monitoring system. RESULTS: Of 149 surgeons responding to the questionnaire, 8% to 12% reported frequent pain in the neck and upper extremities associated with laparoscopic surgery. The ergonomics station demonstrated that the peak and total muscle effort of forearm and thumb muscles were significantly greater (p < 0.01) when the grasping task was performed using the laparoscopic instrument rather than the hemostat. CONCLUSION: These findings indicate that laparoscopic surgical technique is more taxing on the surgeon.

Cumulative Trauma Disorders↗

PubMed1999

Prospects for vaccines of helminth parasites of grazing ruminants.

Defined molecular vaccines for several ruminant heliminth parasites are being pursued at several different laboratories. The most fruitful sources of antigen have been oncosphere surface proteins, excretory/secretory products and integral gut membrane proteins. Nematode gut membrane proteins are unconventional in that they do not come into contact with the host immune response during infection, a feature which brings advantages as well as disadvantages. The genes encoding several of the protective antigens have been cloned, but only in the case of the oncosphere surface proteins has substantial protection been reported with recombinant versions. In addition to the problem of identifying suitable expression systems, issues such as choice of adjuvant and/or the possible use of a vaccine vector have to be solved before molecular vaccines for the economically important helminths can be launched. Of the latter, it seems that vaccines for Haemonchus and Fasciola are the brightest prospects.

Animals↗

PubMed1999

Cloning and characterization of glutamate dehydrogenase (GDH) from the gut of Haemonchus contortus.

Vaccination of lambs with the membrane-bound (S3) thiol-Sepharose binding protein (TSBP) fraction derived from the gut of Haemonchus contortus confers significant protection against homologous challenge. The S3 TSBP peptide profile is dominated by a major protein of ca. 60 kDa which is strongly recognized by antisera from sheep demonstrably protected following immunization with S3 TSBP. In an attempt to identify this protein, sera from protected lambs were employed to screen a lambda gt11 cDNA library of the adult parasite and resulted in the isolation of numerous clones encoding a homologue of the mitochondrial enzyme, glutamate dehydrogenase (GDH). GDH enzyme activity was readily demonstrable in S3 TSBP material and immunolocalization studies showed that the enzyme was localized to the cytoplasm of the parasite's gut. Furthermore, the enzyme appeared to be developmentally regulated, with both GDH mRNA and protein expressed almost exclusively during the blood-feeding parasitic stages.

Amino Acid Sequence↗

PubMed1999

Molecular cloning and characterization of gut-derived cysteine proteinases associated with a host protective extract from Haemonchus contortus.

Cysteine proteinases have been implicated in the protection conferred by vaccination with detergent-soluble extracts of Haemonchus contortus. In the present study, antisera from sheep refractory to Haemonchus challenge following vaccination with a 'proteinase-enriched' Haemonchus gut membrane extract, were employed to screen a cDNA expression library of the adult parasite. This resulted in the isolation of 3 cDNAs (designated hmcp1, 4 and 6) encoding cathepsin B-like cysteine proteinases. Immunocytochemical studies specifically localized the products of these genes to the microvillar surface of the parasite's gut and RT-PCR experiments revealed that these were developmentally regulated, being expressed exclusively during the blood-feeding parasitic stages. In addition, a generic PCR approach was adopted in order to identify the predominant cysteine proteinases in a UK strain of Haemonchus. A panel of 5 cDNAs, including hmcp1 and 4, was amplified in this way. Genomic Southern blot analysis indicated that some of these enzymes were encoded by single-copy genes, whereas others were encoded by multi-copy genes. Subsequent sequence analysis revealed that the proteases identified in this study were distinct from those previously reported in USA strains of the parasite.

Animals↗

PubMed1999

Cloning and characterization of a beta-galactoside-binding protein (galectin) from the gut of the gastrointestinal nematode parasite Haemonchus contortus.

A cDNA encoding a beta-galactoside-binding lectin (galectin) was identified by immunoscreening a Haemonchus contortus cDNA library with antisera from lambs vaccinated with a membrane protein complex (H-gal-GP) derived from the parasites' gut. The cDNA sequence, exhibiting a tandem repeat structure and designated Hco-gal-2, showed significant levels of similarity with galectins from several species of nematode as well as mammalian galectin type 4. Native galectin was preferentially extracted from the H-gal-GP complex and also from an insoluble membrane fraction prepared from adult worms using lactose-agarose affinity chromatography. The affinity-purified material had apparent molecular mass of around 35 kDa with 3 distinct bands visible on SDS-PAGE. All 3 bands were identified as galectins by reaction with antiserum raised to recombinant Hco-GAL-2 on Western blot. To determine whether H. contortus galectins have any protective capacity against infection lambs were vaccinated with affinity-purified galectin and subsequently given a single challenge infection. Vaccination did not confer any protection against infection with H. contortus as judged by faecal egg output or worm counts.

Amino Acid Sequence↗

PubMed1999

Further immunization and biochemical studies with a protective antigen complex from the microvillar membrane of the intestine of Haemonchus contortus.

Immunization of sheep with a protective antigen complex from the intestinal cells of Haemonchus contortus in Freund's adjuvant stimulated individually variable antibody responses but still conferred significant protection against parasite infection. Correlation between antibody concentration and degree of protection was suggestive of antibody being the effector mechanism. The antigen is known as Haemonchus galactose-containing glycoprotein complex (H-gal-GP) because it binds to lectins with a specificity for N-acetyl-galactosamine. Polypeptide composition analysis by polyacrylamide gel electrophoresis indicated an apparent molecular weight of about 1000 kDa and SDS gels revealed four major polypeptides, containing between 2 and 5 disulphide linked subunits, nearly all being glycosylated. N-terminal amino acid sequence was obtained from 12 subunits, ten showing homologies with cDNAs from Haemonchus encoding either pepsin, metalloprotease or cysteine protease-like enzymes. pH optima, inhibitor and various substrate studies confirmed that the native complex possessed proteolytic activities in agreement with the sequence data. Although the cDNAs predicted water soluble enzymes, little of the complex was solubilized from worm membranes without the use of a detergent, such as Triton X-100. It is hypothesized that H-gal-GP is a gut membrane associated multiprotease complex which is involved in the digestion of the blood meal and which can be neutralized by specific antibodies with drastic consequences for the parasite.

Amino Acid Sequence↗

PubMed1999

Immunization with an affinity purified protein extract from the adult parasite protects lambs against infection with Haemonchus contortus.

Young lambs were immunized with membrane protein extracts from adult Haemonchus contortus which had been enriched for cysteine protease activity using Thiol-Sepharose chromatography. The resulting extract comprised several peptides including cysteine as well as serine and metallo-protease activities although the former predominated. A limited number of the peptide components were glycosylated. Following a single challenge infection with the same parasite, vaccinated lambs had substantially reduced faecal egg outputs and final worm burdens (overall means of 77% and 47%, respectively, from three trials) compared with challenge controls. Antibody in sera from vaccinated lambs was almost exclusively bound to the surface of the parasite gut indicating that protection may be mediated by the inhibition of parasite digestion.

Animals↗

PubMed1999

Forty-hertz midlatency auditory evoked potential activity predicts wakeful response during desflurane and propofol anesthesia in volunteers.

BACKGROUND: Suppression of response to command commonly indicates unconsciousness and generally occurs at anesthetic concentrations that suppress or eliminate memory formation. The authors sought midlatency auditory evoked potential indices that successfully differentiated wakeful responsiveness and unconsciousness. METHODS: The authors correlated midlatency auditory evoked potential indices with anesthetic concentrations permitting and suppressing response in 22 volunteers anesthetized twice (5 days apart), with desflurane or propofol. They applied stepwise increases of 0.5 vol% end-tidal desflurane or 0.5 microg/ml target plasma concentration of propofol to achieve sedation levels just bracketing wakeful response. Midlatency auditory evoked potentials were recorded, and wakeful response was tested by asking volunteers to squeeze the investigator's hand. The authors measured latencies and amplitudes from raw waveforms and calculated indices from the frequency spectrum and the joint time-frequency spectrogram. They used prediction probability (PK) to rate midlatency auditory evoked potential indices and concentrations of end-tidal desflurane and arterial propofol for prediction of responsiveness. A PK value of 1.00 means perfect prediction and a PK of 0.50 means a correct prediction 50% of the time (e.g., by chance). RESULTS: The approximately 40-Hz power of the frequency spectrum predicted wakefulness better than all latency or amplitude indices, although not all differences were statistically significant. The PK values for approximately 40-Hz power were 0.96 during both desflurane and propofol anesthesia, whereas the PK values for the best-performing latency and amplitude index, latency of the Nb wave, were 0.86 and 0.88 during desflurane and propofol (P = 0.10 for -40-Hz power compared with Nb latency), and for the next highest, latency of the Pb wave, were 0.82 and 0.84 (P < 0.05). The performance of the best combination of amplitude and latency variables was nearly equal to that of approximately 40-Hz power. The approximately 40-Hz power did not provide a significantly better prediction than anesthetic concentration; the PK values for concentrations of desflurane and propofol were 0.91 and 0.94. Changes of 40-Hz power values of 20% (during desflurane) and 16% (during propofol) were associated with a change in probability of nonresponsiveness from 50% to 95%. CONCLUSIONS: The approximately 40-Hz power index and the best combination of amplitude and latency variables perform as well as predictors of response to command during desflurane and propofol anesthesia as the steady-state concentrations of these anesthetic agents. Because clinical conditions may limit measurement of steady-state anesthetic concentrations, or comparable estimates of cerebral concentration, the approximately 40-Hz power could offer advantages for predicting wakeful responsiveness.

Adult↗

PubMed1999

A virtual instrument ergonomics workstation to measure surgeons' physical stress.

Videoendoscopic (VES) instruments have poor force transmission properties and often require surgeons to employ awkward hand and arm positions. In order to compare the physical workload of laparoscopic surgery to open surgery, we collected long-duration EMG records from the thumb (thenar compartment) of six surgeons performing suturing and knot tying in a training box using both open and VES techniques. EMG signals were acquired using a LabVIEW Virtual Instrument and analyzed using a Modified Exposure Variation Analysis (MEVA) algorithm. Standard EMG indices and the MEVA analysis demonstrated significantly greater amplitude and duration of EMG signals using VES technique compared to open technique. Our results suggest that the use VES techniques requires a greater intensity of physical effort than open surgery techniques.

Algorithms↗

PubMed1998

Craniofacial electromyogram activation response: another indicator of anesthetic depth.

OBJECTIVE: After finding that craniofacial EMG preceding a stimulus was a poor predictor of movement response to that stimulus, we evaluated an alternative relation between EMG and movement: the difference in anesthetic depth between the endpoint of EMG responsiveness to a stimulus and endpoint of movement responsiveness to that stimulus. We expressed this relation as the increment of isoflurane between the two endpoints. METHODS: We measured EMG over the frontalis muscle, over the corrugator muscle, and between the Fp2 and the mastoid process as patients emerged from general anesthesia during suture closing of the surgical incision. Anesthesia was decreased by controlled washout of isoflurane while maintaining 70% N2O, and brain isoflurane concentrations ((C)isoBrain) were calculated. We studied a control group of 10 patients who received only surgical stimulation, and 30 experimental patients who intermittently received test stimuli in addition to the surgical stimulation. Patients were observed for movement responses and EMG records were evaluated for EMG activation responses. We defined an EMG activation response to be a rapid voltage increase of at least 1.0 microV RMS above baseline, with a duration of at least 30 s, in at least one of the three EMG channels. Patient responses to stimuli were classified as either an EMG activation response without a move response (EMG+), a move response without an EMG activation response (MV+), both an EMG activation response and a move response (EMG+MV+), or no response. We defined the EMG+ endpoint to be the threshold between EMG+ response and nonresponse to a stimulus, and estimated (C)isoBrain at this endpoint. We similarly defined the move endpoint and estimated the move endpoint (C)isoBrain. We then calculated the increment of (C)isoBrain at the EMG+ endpoint relative to the move endpoint. MAIN RESULTS: For the 30 experimental patients, the initial response to a test stimulus was an EMG+ in 14 patients (47%), an EMG+MV+ in 12 patients (40%), and a MV+ in 1 patient (3%); no response occurred by the time surgery was completed in 3 patients (10%). No response occurred in 7 of the control patients (70%). Of the 14 patients with an initial EMG+ response to a test stimulus, 9 patients later had a move response. For these 9 patients, the increment of (C)isoBrain between the EMG+ endpoint and move endpoint was 0.11 +/- 0.04 vol%, (mean +/- SD). CONCLUSIONS: Our results suggest that, given the circumstances of our study, an EMG activation response by a nonmoving patient indicates that the patient is at an anesthetic level close to that at which movement could occur. However, because the first EMG activation response may occur simultaneously with movement, the EMG activation response cannot be relied upon to always herald a move response before it occurs. Our results also suggest that EMG responsiveness to a test stimulus may be used to estimate the anesthetic depth of an individual patient.

Adolescent↗