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D W Taylor

Publications and source records attributed to D W Taylor.

At least 145 records · Page 8Linked to original sources

Genetic engineering and a schistosome vaccine.

Early attempts to develop a vaccine against schistosomiasis have been largely empirical, relying on use of live, attenuated larvae or homogenates and extracts of the various life-cycle stages of the parasite. Although some success has been reported using attenuated larvae, by and large the early attempts at vaccination have proved unsuccessful. Recently, we have begun to apply recombinant DNA technology to the problem of schistosome vaccine development. A majority of the evidence pertaining to immunity against schistosome infection suggests that the schistosomula is the primary target of the immune attack. Therefore, it follows that antigens found on the surface of the young larvae would be prime candidates for inclusion in an experimental vaccine. This lecture summarizes work aimed at the identification of schistosomula surface antigens and molecular cloning of their corresponding genes. cDNA clones have been prepared from both egg and adult mRNA. Relevant clones have been identified by "message" selection. This approach relies on identification of relevant antigen precursors among in vitro translation products. At least 5 schistosomula surface-antigen precursors have been recognised among in vitro translation products directed by adult mRNA. Some of these peptides are also encoded by mRNA isolated from eggs. One egg-specific peptide has been cloned and there is evidence that this is immunodiagnostic for Schistoma mansoni infections. A long-term aim is hoped that such molecules would contribute to a schistosome vaccine as well as providing immunodiagnostic reagents.

Animals↗

Isolation and antigen analysis of surface tegument membranes from schistosomula of Schistosoma mansoni.

The outer tegument membranes of Schistosoma mansoni schistosomula have been removed by mechanical disruption in a hypotonic salt solution and partially purified by differential and sucrose density gradient centrifugation. Fractionation was monitored by measuring increase in specific activity of bound [125I]wheat-germ agglutinin (WGA), alkaline phosphatase and calcium-stimulated ATPase. Two-dimensional IEF/SDS polyacrylamide gel electrophoresis was used to analyse the peptide composition of the isolated membranes and to compare and contrast with lactoperoxidase/glucose oxidase surface labelled peptides. At least 35 surface-labelled peptides were resolved on the two-dimensional maps: all were also present in the membrane material recovered from the sucrose gradient. Western blot analysis demonstrated a marked heterogeneity in the antibody response of infected human patients to individual membrane antigens. The antigenic profile of membranes isolated from cercariae, 18 and 96 h schistosomula were compared using Western blots: some minor differences were observed between the three preparations.

Adenosine Triphosphatases↗

Effect of encouragement on walking test performance.

Walking tests, frequently used to document effects of treatment on exercise capacity, have never been standardised. We studied the effects of encouragement on walking test performance in a randomised study that controlled for the nature of the underlying disease, time of day, and order effects. We randomised 43 patients with chronic airflow limitation or chronic heart failure or both to receive or not receive encouragement as they performed serial two and six minute walks every fortnight for 10 weeks. Simple encouragement improved performance (p less than 0.02 for the six minute walk), and the magnitude of the effect was similar to that reported for patients in studies purporting to show beneficial effects of therapeutic manoeuvres. Age and test repetition also affected performance. These results demonstrate the need for careful standardisation of the performance of walking tests, and suggest caution in interpreting studies in which standardisation is not a major feature of the study design.

Chronic Disease↗

Inspection of computer-supported toxicological data submitted to the FDA.

The FDA's Good Laboratory Practice Regulations (GLP) have been formally amended (once) and two formed advisory opinions have been issued. The FDA is now in the process of reviewing the GLPs to comply with both the Regulatory Flexibility Act of 1980 and Executive Order 12291 of 1981. Inspections since 1979 have revealed compliance progress; however, certain areas of the GLPs have been a problem--the definition of "raw data," the documentation process for the maintenance of "raw data," standard operating procedures, and study protocols. The increasing use of computers for supporting toxicology/pathology studies raises several questions concerning the impact of the GLPs on computerized data collection/reporting. This paper will address the above questions and discuss systems, procedures, interpretations, and some unresolved problems, as well as provide practical approaches for internal review of computer-supported nonclinical laboratory studies.

Data Collection↗

Studies on the enhancement of human eosinophil function by mononuclear cell products in vitro.

Previous studies have shown that human mononuclear cells produce during culture an activity that enhances the functional properties of mature eosinophils, including their helminthotoxic potential (Veith & Butterworth, 1983). We have now found that the enhancing activity in mononuclear cell culture supernatants (MCS) is stable to dialysis for 24 h at pH 2 and sensitive to trypsin (0.1 mg/ml). It has a principal pI of 4.4 and possible secondary peaks with pI values of 5.0 and 5.7. The effect of MCS on eosinophils appears to be a general early stimulation of several functions. In comparison with eosinophils incubated in medium only, the proportion of eosinophils with Fc receptors is increased after incubation with MCS for 3.5 h (39 +/- 11% vs 16 +/- 3%) and 7 h (28 +/- 4% vs 17 +/- 3%). The temperature-dependent phase of eosinophil adherence to antibody coated schistosomula is enhanced after 3-4 h. The uptake of deoxyglucose is enhanced after a 1 h pre-incubation with MCS (24 +/- 1 X 10(3) d/min), compared with that of controls (15 +/- 2 X 10(3) d/min). Whether these effects are due to one mediator in MCS awaits further investigation.

Cell Adhesion↗

Analysis of mRNA coding for blood-stage antigens of a rodent malarial parasite, Plasmodium yoelii: mRNA coding for a possible protective antigen purify as poly A-.

We have analyzed mRNA coding for blood-stage antigens of Plasmodium yoelii by using cellfree translation of poly A+ and poly A- RNA in conjunction with immunoprecipitations. Most of the antigens recognized by mouse hyperimmune serum to P. yoelii were coded by poly A+ mRNA ranging in size from 15S to 28S. However, certain P. yoelii antigens, notably those with m.w. greater than 150 kilodaltons (kd), were coded by mRNA that purified as being poly A-. Antigens recognized by a protective monoclonal antibody (McAb) were coded by such operationally poly A- RNA. Three polypeptides apparently coded by different poly A- RNA were immunoprecipitated by this McAb. With the use of another McAb, a poly A+ mRNA of about 19S was identified as coding for a polypeptide of 46 kd synthesized in cellfree translation reactions. The same McAb recognized a 34 kd polypeptide in metabolically labeled polypeptides of P. yoelii. This antigen appeared to be processed in vivo but not in vitro. The observation that some mRNA of P. yoelii purify as being poly A- has significant implications for the construction of cDNA libraries that employ poly A+ mRNA of malarial parasites: if it applies to other species of plasmodia, some potentially important operationally poly A- mRNA may not be represented in such libraries.

Animals↗

Clone banks of cDNA from the parasite Schistosoma mansoni: isolation of clones containing a potentially immunodiagnostic antigen gene.

We have constructed a small vector specifically for blunt-end cloning of fragments of DNA. Both the PvuII site and the EcoRI site allow the detection of recombinants using a simple and inexpensive colour screen. We have used this vector to construct cDNA clone banks from polyadenylated messenger RNA [poly(A)+mRNA] from several life cycle stages of the human parasite Schistosoma mansoni and have identified clones encoding an immunodiagnostic antigen gene by a combination of Southern blotting and mRNA hybrid-selection and in vitro translation. Antibodies against this antigen are only present in patients infected with S. mansoni.

Animals↗

Is there a trade-off between income and health? The case of hypertensive steelworkers in Canada.

It is often assumed that medical treatment creates a trade-off between good health and income; that is, an individual "pays" for good health by an income loss (at least at the early stages of treatment) compared, for example, with the no-treatment choice. We examined empirical data on hypertensive steelworkers in Canada to evaluate whether such a trade-off exists. We found none. Indeed, those who participated in a treatment program to reduce their blood pressure not only gained improved health but also made small income "gains." Our findings thus challenge the validity of the trade-off assumption and call for further studies of its generalizability.

Adult↗

Trial of labor after previous cesarean section: prognostic indicators of outcome.

A cross-sectional analytic survey of 92 patients permitted to attempt vaginal delivery after previous lower segment cesarean section ("trial of scar") is reported. Variables which may predict mode of delivery were assessed. Fifty patients (54.3%) were delivered vaginally; 42 patients (45.7%) had repeat cesarean sections in labor. There were three cases of scar dehiscence (3.2%). There were no maternal or fetal mortality. When the cervix was less than 3 cm dilated at initial examination in labor, 10 of 37 patients (27%) were delivered vaginally, compared to 38 of 55 patients (69%) who were delivered vaginally when the cervix was greater than 3 cm dilated. Assessment of cervical dilatation on admission in labor proved to be the most significant prognostic factor at the onset of labor, with regard to successful vaginal delivery in a patient with a lower segment cesarean section scar.

Cervix Uteri↗

Monoclonal antibodies against surface antigens of schistosomula of Schistosoma mansoni.

Monoclonal antibodies have been produced after fusion of NS-1 murine myeloma cells with spleen cells from mice immunized either by chronic primary infection or with irradiated cercariae: in both cases, animals were challenged with liver cercariae 7 days before fusion. The initial cultures were screened for anti-schistosomular antibodies both by a radioimmunoassay with whole schistosomulum extracts and by immunofluorescence. There was no correlation between the two techniques and subsequent screening was carried out by immunofluorescence. Cloning was carried out in soft agar and 7 cloned cell lines, from 5 initial cultures, were selected for detailed study. Products of 6 of these 7 lines were monoclonal, as judged by isoelectricfocusing of [35S]methionine-labelled supernatant fluids, and their binding to live schistosomula was specific. None of the antibodies showed detectable activity in mediating eosinophil- or complement-dependent damage to schistosomula in vitro. However, 2 antibodies were successfully used to isolate surface proteins with an apparent molecular weight of 24 000 on SDS-polyacrylamide gel electrophoresis.

Animals↗

Toxoplasmosis in immunoglobulin M-suppressed mice.

Mice challenged with a pathogenic strain of Toxoplasma gondii develop fatal infections. However, if such mice are initially treated with sulfadiazine (SD), they develop immunity and survive with chronic infections. The role of antibody (Ab) in establishing protective immunity against acute parasitemias and in maintaining chronic infections was investigated using B-cell-deficient (immunoglobulin M-suppressed), T-cell-deficient (athymic), and normal BALB/c mice. All mice not receiving SD treatment rapidly died (mean 7.5 days) after infection, but the majority (80%) of intact mice developed immunity during SD treatment and survived for over 5 months with chronic toxoplasmosis. Athymic mice rapidly died (mean 6.0 days) after the removal of SD treatment. Although all SD-treated immunoglobulin M-suppressed mice eventually died, they lived considerably longer (18 to 83 days) in the complete absence of antitoxoplasma Ab than unprotected mice (7 to 9 days). Histopathological sections of liver, lung, brain, and other tissues showed that toxoplasma organisms gave rise to fatal lesions in all nonsurviving animals. The injection of Ab into acutely infected and athymic mice imparted no protection, but transfer of antitoxoplasma Ab (titer greater than 1:8,000) to immunoglobulin M-suppressed mice after SD treatment resulted in elimination of the parasites in 50% of the mice. Results of this study suggest that Ab may not be decisive in acute infections, but may be important in controlling long-term toxoplasmosis.

Animals↗