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W D Smith

Publications and source records attributed to W D Smith.

At least 19 recordsLinked to original sources
PubMed2006

Developments and hurdles in generating vaccines for controlling helminth parasites of grazing ruminants.

As a direct consequence of rising drug resistance among common nematodes of grazing animals, efforts toward state-of-the-art vaccine development have clearly intensified in recent years, fuelled primarily by the advent of newer technologies in gene discovery, by advancements in antigen identification, characterisation and production. In this regard, it is appropriate to review progress that has been made in generating helminth vaccines and in particular, vaccines against common nematodes of production animals for consumption. In like manner, it is prudent to evaluate barriers that have hindered progress in the past and continue to present obstacles that must be solved when utilizing and depending on host immunity to attenuate parasitic infections.

Animals↗
PubMed2006

Molecular characterization of a family of metalloendopeptidases from the intestinal brush border of Haemonchus contortus.

Substantial protection against the economically important parasitic nematode Haemonchus contortus has been achieved by immunizing sheep with a glycoprotein fraction isolated from the intestinal membranes of the worm (H-gal-GP). Previous studies showed that one of the major components of H-gal-GP is a family of at least 4 zinc metalloendopeptidases, designated MEPs 1-4. This paper describes aspects of the molecular architecture of this protease family, including the proteomic analysis of the MEP fraction of the H-gal-GP complex. These enzymes belong to the M13 zinc metalloendopeptidase family (EC 3.4.24.11), also known as neutral endopeptidases or neprilysins. The sequences of MEPs 1 and 3 suggested a typical Type II integral membrane protein structure, whilst MEPs 2 and 4 had putative cleavable signal peptides, typical of secreted proteins. Proteomic analysis of H-gal-GP indicated that the extracellular domain of all 4 MEPs had been cleaved close to the transmembrane region/signal peptide with additional cleavage sites mid-way along the polypeptide. MEP3 was present as a homo-dimer in H-gal-GP, whereas MEP1 or MEP2 formed hetero-dimers with MEP4. It was found that expression of MEP3 was confined to developing 4th-stage larvae and to adult worms, the stages of Haemonchus which feed on blood. MEP-like activity was detected in the H-gal-GP complex over a broad pH range (5-9). Since all 4 MEPs must share a similar microenvironment in the complex, this suggests that each might have a different substrate specificity.

Amino Acid Sequence↗
PubMed2005

An immunogenic cathepsin F secreted by the parasitic stages of Teladorsagia circumcincta.

Teladorsagia circumcincta is a common, pathogenic abomasal nematode of sheep. In order to improve disease control in parasite isolates resistant to several anthelmintics, alternative methods must be sought. Sheep develop acquired immunity to T. circumcincta so vaccination is a valid option for control. For this reason, we are investigating parasite excretory/secretory products for molecules, which have potential to invoke protective immunity against T. circumcincta. Here, we describe experiments in which we identified a novel, immunogenic cathepsin F secreted by L4 T. circumcincta. This protease, initially identified by mass spectrometry analysis, is the most abundant molecule in excretory/secretory products released in vitro by T. circumcincta harvested at 5, 6 or 9 days p.i. and is a target of specific, local IgA responses in sheep which are immune to challenge infection. The full-length cDNA encoding this secreted protease was isolated. Sequence and phylogenetic analyses indicated that the protease (designated T. circumcincta cathepsin F-1, Tci-CF-1) belongs to the cathepsin F class and exhibits greatest identity (>60%) to expressed sequence tags present in the Ostertagia ostertagi and Haemonchus contortus expressed sequence tag databases. Tci-CF-1 also displays high identity to hypothetical proteins identified in the genomes of Caenorhabditis elegans and Caenorhabditis briggsae, both proteins having been described as cathepsin F enzymes. Specific inhibitor binding assay of larval excretory/secretory products confirmed the classification of this excretory/secretory component as a cathepsin F. Reverse transcription-PCR studies indicated that Tci-cf-1 is developmentally regulated and is particular to the host parasitic stages of T. circumcincta. The abundance, immunogenicity and temporal expression pattern of Tci-CF-1 make this a potential vaccine candidate for teladorsagiosis.

Amino Acid Sequence↗
PubMed2005

Protection studies with a globin-enriched protein fraction of Ostertagia ostertagi.

The protective capacity of an adult stage Ostertagia ostertagi globin antigen was tested in four vaccination experiments in cattle. In a preliminary experiment, calves were vaccinated three times intraperitoneally with 250 microg globin in Freund's adjuvant and challenged with a trickled infection of 25,000 infective larvae. In three subsequent field studies, calves were vaccinated twice or three times intramuscularly with 80-100 microg globin in Quil A and challenged with a natural gastrointestinal nematode infection on pasture. Higher globin-specific antibody levels were detected in the vaccinated calves than in the control animals in all vaccine trials. In the preliminary experiment, geometric mean cumulative egg counts in the globin group were reduced by 52% and total worm burdens were reduced by 28%, compared to the controls. In the first field trial cumulative faecal egg counts were reduced by 63% in the vaccinated calves. However, the reduction in faecal egg output in these two experiments was not statistically significant and no reduction in faecal egg counts was observed in the vaccinated animals in the two last field trials. In conclusion, vaccination of calves with O. ostertagi globin resulted in highly variable protection levels after challenge infection.

Adjuvants, Immunologic↗
PubMed2005

Protection induced by vaccinating sheep with a thiol-binding extract of Haemonchus contortus membranes is associated with its protease components.

Previous work has shown that a protein extract enriched for cysteine protease activity (TSBP) prepared from adult Haemonchus contortus using thiol sepharose affinity chromatography confers substantial protection against a single challenge infection. TSBP comprised proteases and other proteins. Here, TSBP were further fractionated using anion-exchange chromatography and fractions pooled on the basis of containing (1) protease activity, (2) a prominent non-protease peptide and (3) material which did not bind to the column. A protection trial showed that only the protease-enriched material conferred protective immunity and this was comparable to that observed in a TSBP-immunized positive control group. Immunization stimulated a marked IgG response with the IgG2 isotype predominating.

Animals↗
PubMed2003

Metalloendopeptidases from the intestinal brush border of Haemonchus contortus as protective antigens for sheep.

Substantial protection against the economically important nematode Haemonchus contortus has been achieved by immunizing sheep with a glycoprotein fraction isolated from the intestinal membranes of this parasite. This fraction has been termed Haemonchus galactose-containing glycoprotein complex (H-gal-GP) since it was originally isolated through its selective binding to lectins with a specificity for N-acetylgalactosamine. A major component of this highly protective antigen complex is a family of four zinc metalloendopeptidases, designated MEPs 1-4. Various combinations of these MEPs were evaluated in immunization-challenge trials in sheep. In two experiments a combination of all four MEPs, separated from the rest of the complex by gel filtration in 8 m urea, significantly reduced H. contortus egg counts by 45 and 50%, an effect not significantly different from that conferred by 8 m urea treatment of H-gal-GP itself. Similarly, MEP3 alone or MEPs 1, 2 and 4 in combination, electroeluted from the complex following SDS gel electrophoresis, each reduced egg counts by some 33%. The MEPs are therefore protective components of H-gal-GP and from previously published findings, it appears that MEP3 is the most effective member of this metalloendopeptidase family. However, there was no significant protection when sheep were immunized with fully reduced and denatured H-gal-GP or with bacterially expressed recombinant forms of MEP 1 or the principal domains of MEP3, suggesting that conformational epitopes on the MEPs are required for immunity.

Animals↗
PubMed2002

Development of a protocol for monitoring status and trends in forest soil carbon at a national level.

The national Forest Health Monitoring (FHM) program requires protocols for monitoring soil carbon contents. In a pilot study, 30 FHM plots loblolly shortleaf (Pinus taeda L./Pinus echinata Mill.) pine forests across Georgia were sampled by horizon and by depth increments. For total soil carbon, approximately 40% of the variance was between plots, 40% between subplots and 20% within subplots. Results by depth differed from those obtained by horizon primarily due to the rapid changes in carbon content from the top to the bottom of the A horizon. Published soil survey information overestimated bulk densities for these forest sites. The measurement of forest floor depths as a substitute to sampling did not provide reliable estimates of forest floor carbon. Precision of replicate samples was approximately 10-30% for field duplicates and 5-10% for laboratory duplicates. Based on national indicator evaluation criteria, sampling by depth using bulk density core samplers has been recommended for national implementation. Additional procedures are needed when sampling organic soils or soils with a high percentage of large rock fragments.

Carbon↗
PubMed2002

Attempts to immunize sheep against the scab mite, Psoroptes ovis.

Adult Psoroptes ovis were successively extracted in saline, 1% Tween, 1% Triton and 8 m urea +0.1% CHAPS. The Triton extract was separated into fractions which did or did not bind to ConA lectin. Using QuilA as adjuvant, both Triton fractions and the saline, Tween and urea extracts were tested separately as candidate protective antigens against a P. ovis challenge infestation in sheep. All induced circulating antibody responses, but the saline and Tween extracts also stimulated significant protective effects in those sheep which developed active disease. Compared to control sheep injected with adjuvant only, these immunized animals had more than two and seven-fold reductions in mean lesion areas and mite numbers, respectively.

Animals↗
PubMed2001

An ergonomic study of the optimum operating table height for laparoscopic surgery.

BACKGROUND: Laparoscopic surgery requires the use of longer instruments than open surgery, thus changing the relation between the height of the surgeon's hands and the desirable height of the operating room table. The optimum height of the operating room table for laparoscopic surgery is investigated in this study. METHODS: Twenty-one surgeons performed a two-handed, one-fourth circle cutting task using a laparoscopic video system and laparoscopic instruments positioned at five instrument handle heights relative to subjects' elbow height (-20, -10, 0, +10, and +20 cm) by adjusting the height of the trainer box. Subjects rated the difficulty and discomfort experienced during each task on a visual analog scale. Skin conductance (SC) was measured in Micromhos via paired surface electrodes placed near the ulnar edge of the palm of the right (cutting) hand. The mean electromyographic (EMG) signal from the right deltoid and trapezius muscles was measured. Arm orientation was measured in three dimensions using a magnetometer/accelerometer. Signals were acquired using analog circuitry and digitally sampled using a National Instruments DAQCard 700 connected to a Macintosh PowerBook 5300c running LabVIEW software. Statistical analysis was carried out by analysis of variance and post hoc testing. RESULTS: Statistically significant changes were found in the subjective rating of discomfort (p <0.002), deltoid EMG (p <0.0006), trapezius EMG (p <0.0001), and arm elevation (p <0.0001) between instrument handle heights. SC values and task times did not change significantly. Discomfort and difficulty ratings were lowest when instrument handles were positioned at elbow height. EMG values and arm elevation all decreased with lower instrument height. CONCLUSION: This study suggests that the optimum table height for laparoscopic surgery should position the laparoscopic instrument handles close to surgeons' elbow level to minimize discomfort and upper arm and shoulder muscle work. This corresponds to an approximate table height of 64 to 77 cm above floor level. A redesign of current operating room tables may be required to meet these ergonomic guidelines.

Equipment Design↗
PubMed2001

Vaccination against gastrointestinal nematode parasites of ruminants using gut-expressed antigens.

To date, proteins isolated from the surface of the gut of gastrointestinal nematodes, particularly Haemonchus contortus, have generally proved to be useful protective antigens and several are being progressed towards recombinant protein-based vaccines. This paper describes the properties of some of the most promising antigens and summarises their performance in laboratory and field based trials. The antigens described include contortin, H11, H-gal-GP, GPI and cysteine proteinases. In addition, the discussion addresses the utility of selected antigens to protect against co-infecting nematode species such as Teladorsagia circumcincta and against related nematode infections such as Ostertagia ostertagi in cattle.

Animals↗
PubMed2001

Preliminary observations on the potential of gut membrane proteins of Haemonchus contortus as candidate vaccine antigens in sheep on naturally infected pasture.

In a trial lasting 11 months in South Africa, faecal egg counts and haematocrits of sheep vaccinated with gut membrane proteins of adult Haemonchus contortus were compared with unvaccinated controls grazing pasture contaminated with the parasite. Vaccination reduced egg output by >82% on average during one 4 month period of the trial and simultaneously significantly reduced the degree of anaemia and deaths due to haemonchosis. Although vaccine immunity was not sufficiently long lasting to prevent a surge in egg output which occurred after the onset of a period of irrigation, re-vaccinating the sheep at this point cleared their newly acquired infection and rapidly restored protection to approximately the level observed beforehand. It was clear that a vaccine based on parasite gut membrane proteins could offer substantial benefits in the control of natural haemonchosis.

Animals↗
PubMed2001

The effect of laparoscopic instrument working angle on surgeons' upper extremity workload.

BACKGROUND: Laparoscopic surgery may be kinder to the patient, but it is more demanding on the surgeon. Fixed trocar positions often require the surgeon to work with instruments at awkward angles to their body. We studied the effect of horizontal and vertical laparoscopic instrument working angle on the surgeon's thumb, forearm, and shoulder muscle work. METHODS: Electronyographic (EMG) signals were collected from the thenar compartment (TH), flexor digitorum superficialis (FDS), and deltoid (DEL) muscles of the dominant arm of eighth surgeons while they were closing a standard pistol-grip disposable laparoscopic grasper against a fixed resistance of 3 N. With the aid of a special testing bench, the instruments' position was randomly changed among 15 degrees, 45 degrees, and 75 degrees of horizontal angulation relative to the surgeons' sagital plane, and 15 degrees, 45 degrees, and 75 degrees degrees of vertical angulation relative to a horizontal plane. EMG signals were rectified and smoothed using analogue circuitry and digitally sampled at l0 Hz using a National Instruments DAQCard-700 connected to a Macintosh PowerBook 5300c running LabVIEW software. Statistical analysis was carried out by analysis of variance (ANOVA). RESULTS: The effects of vertical and horizontal working angles on the muscle effort were as follows: TH (horizontal, N.S.; vertical, N.S.), FDS (horizontal, p 45 degrees to the surgeon's sagital plane significantly increases the workload of the flexor digitorium superficialis and deltoid muscles. The deltoid muscle is also adversely affected by vertical angulation of the instrument. The instrument working angle has no effect on the thenar muscles. Whenever possible, laparoscopic surgeons should strive to place their instruments and trocars so as to minimize extreme horizontal or vertical displacement of their hands away from a resting position of comfort.

Arm↗
PubMed2001

Performing laparoscopic surgery is significantly more stressful for the surgeon than open surgery.

BACKGROUND: The awkward visual and physical interface of video-endoscopic surgery (VES) has been shown to increase the physical workload of the surgeon, yet there is a lack of objective information on the mental effort and stress demanded by VES techniques. This study addresses the hypothesis that VES is more stressful than open surgery using an ergonomic analysis of surgical tasks in a laboratory setting. METHODS: A portable ergonomic work station was developed using a software Virtual Instrument (VI) interfaced with electronic hardware to compare the mental workload of 28 surgeons. The task was knot tying. The independent variable was work condition: rest, open technique, or VES technique. Dependent variables were tonic skin conductance level (SCL), electrooculogram (EOG), and subjective reports of concentration (CON) and stress (STR). Statistical analysis used nonparametric methods. RESULTS: Subjects tied fewer knots using the VES technique (p < 0.05). The SCL increased progressively from rest to the open task to the VES task (p < 0.05), correlating with the subjects' reported increase in mental stress level (p < 0.05). Eye blinks decreased from rest to the open task (p < 0.05), consistent with the subjects' reported increase in level of mental concentration. From the open to the VES task, eye blinks increased (p < 0.05), as would be expected given the greater demands of the VES task. Experienced subjects demonstrated less variability in SCL levels across tasks. CONCLUSIONS: VES technique requires greater concentration and places greater mental stress on surgeons than to open surgery. More experience with VES may decrease this effect. Studies are needed to improve the human-technology interface the stress.

Attention↗
PubMed2001

An ergonomic evaluation of surgeons' axial skeletal and upper extremity movements during laparoscopic and open surgery.

BACKGROUND: Many surgeons have complained of fatigue and musculoskeletal pain after laparoscopic surgery. We evaluated differences in surgeons' axial skeletal and upper extremity movements during laparoscopic and open operations. METHODS: Five surgeons were videotaped performing 16 operations (8 laparoscopic and 8 open) to record their neck, trunk, shoulder, elbow, and wrist movements during the first hour of surgery. We also compared postprocedural complaints of pain, stiffness, or numbness between the two groups. RESULTS: Compared with surgeons performing open surgery, surgeons performing laparoscopic surgery exhibited less lateral neck flexion; less trunk flexion; more internal rotation of the shoulders; more elbow flexion; more wrist supination and wrist ulnar and radial deviation. There was a trend of more shoulder stiffness after laparoscopic operations than after open operations. CONCLUSIONS: Laparoscopic surgery involves a more static posture of the neck and trunk, but more frequent awkward movements of the upper extremities than open surgery. Ergonomic changes in the operating room environment and instrument design could ease the physical stress imposed on surgeons during laparoscopic operations.

Adult↗
PubMed2001

Cloning and expression of cystatin, a potent cysteine protease inhibitor from the gut of Haemonchus contortus.

A cDNA encoding a cysteine protease inhibitor (cystatin) was identified by immunoscreening a Haemonchus contortus cDNA library with antisera from lambs vaccinated with a protective membrane protein complex (H-gal-GP) derived from the gut of the parasite. The cDNA sequence, designated Cys-1, showed significant levels of similarity with cystatins from several species of nematode as well as with human cystatin. Recombinant H. contortus cystatin was expressed in Escherichia coli in a soluble and functionally active form, which proved to be a potent inhibitor of both mammalian cathepsin B and native H. contortus cysteine proteases. Immunolocalization studies using antisera raised against recombinant H. contortus cystatin showed that the inhibitor was predominantly expressed in the cytoplasm of intestinal cells. To determine whether H. contortus had any protective capacity against infection, lambs were vaccinated with the recombinant molecule and subsequently given a single challenge infection. Although vaccination did not confer any protection against infection with H. contortus, as judged by faecal egg output or worm counts, cystatin will be a valuable tool in the analysis of the function of the cysteine proteases which are the subject of on-going study as potential vaccine components.

Amino Acid Sequence↗
PubMed2001

Cross-protection studies with gut membrane glycoprotein antigens from Haemonchus contortus and Teladorsagia circumcincta.

Gel profiles of the peanut and ConA lectin binding integral membrane glycoproteins of Teladorsagia circumcincta and Haemonchus contortus were compared and found to be considerably different. However, some of the Teladorsagia polypeptides were recognized by antisera specific for Haemonchus amino-, metallo- or aspartyl peptidases, enzymes which are known to be protective antigens for that parasite. As expected, an experimental vaccine containing these Haemonchus proteases was extremely effective against homologous challenge, reducing egg and worm counts by more than 99% and 92%, respectively, but it did not provide any useful cross-protection against either T. circumcincta, Trichostrongylus axei or Cooperiaoncophora. A reciprocal experiment, where sheep were immunized with the equivalent glycoproteins from T. circumcincta, showed that, while they were not protected against homologous challenge, there was some cross-protection against Haemonchus as measured by a significant reduction in worm egg output.

Animals↗