Biomedical subjects

D D Heath

Publications and source records attributed to D D Heath.

At least 19 recordsLinked to original sources

PubMed1998

Mytilus protamine-like sperm-specific protein genes are multicopy, dispersed, and closely associated with hypervariable RFLP regions.

Protamine-like sperm packaging proteins replace somatic histones during spermatogenesis, and although the proteins have been well-characterized in many marine invertebrate species, little is known of the arrangement of the genes. The research described here was designed to determine the sequence and structure of the protamine-like PL-III (or "phi 1") gene in marine mussels (Mytilus spp). The PL-III sequence was found to be extremely variable not only among the closely related Mytilus species, but also within species and populations. The variation observed among eight PL-III sequences from a single individual indicated that PL-III was probably multiple-copied. Southern analysis confirmed that PL-III, and another protamine-like gene, (PL-II), were multicopy and dispersed, as well as associated with a hypervariable element. Some PL-III genes are also arranged in nontandem clusters, and the spacer regions are probably the source of the hypervariable nature of the Southern blots. The arrangement of the protamine-like genes in Mytilus appears to be closer to that reported for histones than protamines; however, their association with a hypervariable element is novel.

Amino Acid Sequence↗

PubMed1997

Vaccination against echinococcus in perspective.

Vaccination of intermediate hosts will effectively reduce the prevalence of human echinococcosis. The rate of progress can be enhanced by concurrent tactical treatment of definitive hosts. The oncosphere of Echinococcus granulosus contains a specific protein which when expressed on the oncosphere surface will combine with specific antibody In the presence of complement, damage is caused to the plasma membrane, resulting in failure of the oncosphere to develop into a metacestode. Immunisation of sheep with oncosphere secretions or with an oncosphere homogenate results in 99% resistance to a challenge infection of the sheep with E. granulosus eggs. Immunisation of sheep with the specific purified protein results in 92% resistance. The gene coding for the protein has been isolated from an E. granulosus oncosphere cDNA library and expressed in E.coli as a fusion protein with glutathione-S-transferase. A monoclonal antibody to an epitope on the E. granulosus portion of the molecule is able to bind to the surface of oncospheres and to prevent their development to metacestodes in vitro. Immunisation of sheep with the fusion protein has given 97 and 98% resistance to a challenge infection in two separate trials. The shelf-life of formulated vaccine is at least 12 months, and the immunity generated by two injections given 1 month apart persists for at least 12 months. A homologue of the gene has been identified in E. multilocularis. The vaccine has the potential to be used as a tool for control of transmission of E. granulosus through its natural intermediate hosts (particularly domestic ungulates) as part of hydatid control programmes. Traditional methods of hydatid control (anthelmintic treatment of dogs) require regular treatment within the prepatent period for prolonged periods (20-30 years) to achieve satisfactory control, and often sociological factors to do with dog-feeding, dog ownership and dog-control prevent such an achievement. By controlling the disease in both the definitive host and the intermediate host, the shortcomings of either procedure can be overcome, resulting in a cost-effective desired result within a practical time period.

Animals↗

PubMed1997

Pathological studies of wobbly possum disease in New Zealand brushtail possums (Trichosurus vulpecula).

Following an outbreak of wobbly possum disease in a colony of brush tail possums (Trichosurus vulpecula), the disease was established experimentally in captive possums by inoculating the animals intraperitoneally with tissue homogenates. Crude tissue homogenates of liver remained infectious after freezing at -75 degrees C or filtration through a 0.22 micron filter. The disease was characterised by docility, incoordination, loss of balance and wasting. Fifteen of 16 infected animals had to be euthanased owing to the severity of clinical signs. Cachexia was the only change observed postmortem. Histology revealed widespread perivascular infiltrations with plasma cells and lymphocytes which were severe in the liver and kidney and moderate to mild in a variety of other tissues, including skeletal and cardiac muscle. Changes in the brain consisted of a mild to moderate mononuclear perivascular cuffing. Most of the animals had small to large numbers of circulating nucleated red blood cells and eosinopenia when they were euthanased. There was a consistent decrease in serum albumin concentration and an increase in serum globulins, which resulted in a decreased albumin:globulin ratio. Virus-like particles were observed in preparations of liver from two animals; they appeared to be spherical or icosahedral and were 45 nm in diameter.

Animals↗

PubMed1997

The early stages of Echinococcus granulosus development.

The paper points out the essential aspects of host/parasite relationship during the development of oncospheres (ONC) of Echinococcus granulosus into an early cyst. Secretions of the penetration glands in the hatched ONC causes lysis of host tissue during penetration of the activated ONC. It also protects the parasite against the host's immune response while developing the laminated layer. The microvilli, compressed under the plasma membrane in the hatched ONC, are uplifted in the activated larvae; they increase in number and size and are substituted by short and truncated microtriches in metacestodes 3 days old. At that time appears the first lamination of the laminated layer that surrounds the metacestodes as an electron-dense matrix composed of fine microfibrillated material and remnants of sloughed microvilli. The second lamination appears by day 6-8 and is more electron-dense than the first one. The laminated layer is of parasite origin, formed of a series that emanates from the germinal membrane. It is suggested that Vg secretory vesicles, that are elaborated in the perikaryon of the germinal zone and continuously carried to the syncytium via the microtubular-cytoskeleton, are responsible for the laminated layer formation and are involved in initial evasion of the immune response of the host. The cyclical production of laminations could be necessary to create layers that can ultimately be sloughed off as the cyst grows and serve to divert the host cellular response to the parasite.

Animals↗

PubMed1996

Identification and cDNA cloning of two novel low molecular weight host-protective antigens from Taenia ovis oncospheres.

Oncosphere antigens of Taenia ovis were solubilised in sodium dodecyl sulphate and separated by electrophoresis in polyacrylamide gels (SDS-PAGE). Antigen-containing gel fractions cut from the region covering 18-12 kDa were shown to be highly immunogenic in sheep challenge experiments. Specific antisera against 2 candidate antigens at 18 and 16 kDa were used to screen a cDNA library prepared from T. ovis oncosphere mRNA. Recombinant proteins selected with antibody to the 16 and 18 kDa native antigens were expressed as GST fusion proteins. Vaccination trials using either of the 2 fusion proteins To16.17-GST and To18-GST, revealed that each was capable of inducing high levels of immunity in sheep against challenge infection with T. ovis eggs. Antibodies induced by vaccination with the recombinant antigens reacted specifically with their respective 18 or 16 kDa native oncosphere antigens. There was no apparent homology between the T. ovis cDNA coding for To18 and To16.17, or with another host-protective antigen, To45W, described previously. These additional host-protective antigens should prove a valuable adjunct to To45W and permit the development of effective vaccination strategies.

Amino Acid Sequence↗

PubMed1996

Neurological complications of ankle arthroscopy.

A retrospective review of the first 612 patients undergoing consecutive ankle arthroscopy in the practices of two experienced arthroscopists was under-taken. All inpatient records, outpatient charts, and operative reports were reviewed. Indications for surgery included pain, swelling, locking, and instability that failed to respond to nonoperative management. The results of our investigation revealed an overall complication rate of 9.0% (55 complications). There were 27 neurological complications (4.4% of all arthroscopies) accounting for 49.1% of the complications noted. Specifically, the superficial peroneal nerve was injured in 15 cases, the sural nerve in 6, the saphenous nerve in 5, and the deep peroneal nerve in 1. All nerve injuries occurred through direct injury by portal or distractor pin placement. No cases of neurological injury caused by tourniquet compression or compartment syndrome were seen. Also, 1 case of reflex sympathetic dystrophy was identified.

Adolescent↗

PubMed1996

Antigenic polypeptides of Echinococcus granulosus oncospheres and definition of protective molecules.

Immunoblotting and in vitro oncosphere-killing were used to identify a putative protective molecule in Echinococcus granulosus mature oncospheres. A range of sera from sheep that had been shown to be protected against E. granulosus, and from those that were not, were tested. The sera used were obtained from sheep hyperimmunized with E. granulosus oncospheres, or immunized with oncosphere non-denatured extract, with immature oncosphere extract or with denatured extracts of oncospheres. Results indicated the involvement of native antigens of 23, 25, 30, 34 and 40 kDa in the protective response to E. granulosus infection. The rapid appearance of antibodies to the 23, 25 kDa antigens, their association with early onset of protection and in vitro oncosphere lysis by affinity-purified antibodies obtained from these fractions, indicated that these antigens contained protective epitopes. Final confirmation was provided by immunization of sheep with fractions prepared by preparative SDS/PAGE, and challenge infection. Only the fraction containing the 23 and 25 kDa molecules was able to stimulate protection. Antisera against this pair of molecules should provide a useful probe for screening an E. granulosus oncosphere cDNA library to identify clones expressing protective molecules.

Animals↗

PubMed1996

Immune responses associated with protection in sheep vaccinated with a recombinant antigen from Taenia ovis.

This paper describes the evaluation of the protective antibody response of sheep to vaccination against Taenia ovis infection with a defined recombinant antigen (45W). Sera from 181 vaccinated sheep, collected prior to experimental challenge with T.ovis, were assessed for 45W specific IgA, IgG, IgG1, IgG2 and IgM levels and these results correlated with protection data. There were significant relationships (P < 0.001) between IgG, IgG1 and IgG2 titres and protection. Serum IgA levels did not correlate with protection and there were no significant levels of 45W specific IgM detected. Killing of several other taeniid cestodes has been shown to be complement mediated and the findings in this study are consistent with the involvement of this immune mechanism in 45W vaccinated sheep. A comparison of the adjuvants used in this study (saponin and oil in water) demonstrated that whereas both adjuvants stimulated the production of similar levels of 45W specific IgG1, the IgG2 response was significantly higher in sheep vaccinated with oil adjuvant.

Adjuvants, Immunologic↗

PubMed1996

Vaccination against hydatidosis using a defined recombinant antigen.

Echinococcus granulosus is the causative agent of hydatid disease in humans and animals. Natural transmission of the parasite occurs between dogs as definitive hosts and animal intermediate hosts. There is an urgent need for improved methods to control the parasite's transmission. Here we describe the development of a vaccine based on a cloned recombinant antigen from the parasite egg (oncosphere). Sheep-vaccinated with the antigen, designated EG95, are protected (mean 96-98%) against hydatidosis developing from an experimental challenge infection with E. granulosus eggs. The vaccine will provide a valuable new tool to aid in control of transmission of this important human pathogen. It also has the potential to prevent hydatid disease directly through vaccination of humans.

Amino Acid Sequence↗

PubMed1996

Ecological interaction between sympatric Mytilus species on the west coast of Canada investigated using PCR markers.

M. californianus is the dominant marine mussel species on exposed rocky shores, while M. trossulus is usually the dominant mussel species in more sheltered waters on the west coast of North America. Since these species are physical indistinguishable when small (< 10.0 mm), we developed two polymerase chain reaction (PCR) -based markers to discriminate between them. Using these markers, we identified mussels taken from an exposed coast (n = 114), a sheltered harbour (n = 80), and an upper-intertidal pool (n = 42) on the west coast of Vancouver Island, British Columbia, Canada. M. californianus were found only on the open coast. Small M. trossulus (< 20.0 mm) were common to all three sample sites, but were extremely rare at larger sizes (> 20.0 mm) on the open coast. Our results indicate that M. californianus are excluded from sheltered waters via early life factors, while M. trossulus are excluded from the open coast due to mortality later in life.

Animals↗

PubMed1995

Selective intra-arterial infusion of high-dose cisplatin in patients with advanced head and neck cancer results in high tumor platinum concentrations and cisplatin-DNA adduct formation.

A group of 23 patients with advanced head and neck cancer were treated with highly selective intra-arterial (IA) cisplatin 150 mg/m2 delivered rapidly through microcatheters. The systemic effects of cisplatin were neutralized by concurrent administration of sodium thiosulfate. Two-to-threefold higher tumor platinum contents were detected in tumor biopsies after selective IA cisplatin administration compared to historical controls (treated with 100 mg/m2 IA). Cisplatin-induced DNA modification in human tumor biopsies was quantitated using the antiserum NKI-A59. High levels of cisplatin DNA adducts were detected which correlated linearly with the tumor platinum content (r2 = 0.62). The addition of radiotherapy to this high dose intensity cisplatin treatment resulted in a 92% complete response (CR) rate (12 of 13 patients achieved a CR). Since no difference in tumor platinum content was detected between patients receiving or not receiving radiotherapy (13 and 10 patients, respectively), but the response rate was substantially different (12 CR and 1 partial response with radiotherapy versus 6 partial and 4 non-responders without radiotherapy), these data suggest that the high platinum levels achieved by selective IA infusion were sufficient to produce enough interaction with radiotherapy to cause a 92% CR rate. Whether this interaction is additive or synergistic is as yet unclear.

Antineoplastic Agents↗

PubMed1995

Taenia ovis recombinant vaccine--'quo vadit'.

Several years have elapsed since the publication by Johnson et al. (1989) of the cloning of a recombinant antigen from the cestode parasite Taenia ovis which stimulated high levels of protective immunity in sheep. A great deal of subsequent research and development was necessary to bring the fledgling vaccine to the point of being a registered commercial product. The results of these subsequent studies are dealt with briefly in this paper, including the results of field trials. The T. ovis vaccine was registered by the New Zealand Animal Remedies Board in February 1994. Where then is the commercial product? This paper gives a background to market problems which have emerged through the politics (and realities) of the NZ T. ovis control campaign. It serves as notice that the best science dedicated to producing vaccines or products for parasitic, or other, diseases often faces significant hurdles in the real world of commerce and politics.

Animals↗

PubMed1995

Cross-resistance between cisplatin, antimony potassium tartrate, and arsenite in human tumor cells.

Cross-resistance between cisplatin (DDP) and metalloid salts in human cells was sought on the basis that mechanisms that mediate metalloid salt cross-resistance in prokaryotes are evolutionarily conserved. Two ovarian and two head and neck carcinoma cell lines selected for DDP resistance were found to be cross-resistant to antimony potassium tartrate, which contains trivalent antimony. The DDP-resistant variant 2008/A was also cross-resistant to arsenite but not to stibogluconate, which contains pentavalent antimony. A variant selected for resistance to antimony potassium tartrate was cross-resistant to DDP and arsenite. Resistance to antimony potassium tartrate and arsenite was of a similar magnitude (3-7-fold), whereas the level of resistance to DDP was greater (17-fold), irrespective of whether the cells were selected by exposure to DDP or to antimony potassium tartrate. In the resistant sublines, uptake of [3H]-dichloro(ethylenediamine) platinum(II) was reduced to 41-52% of control, and a similar deficit was observed in the accumulation of arsenite. We conclude that DDP, antimony potassium tartrate, and arsenite all share a common mechanism of resistance in human cells and that this is due in part to an accumulation defect.

Anions↗

PubMed1995

A defined antigen for the serodiagnosis of Taenia ovis infections in dogs.

The evaginated scolex of Taenia ovis secretes an antigen complex into defined culture medium that has been used to develop a cestode-specific enzyme-linked immunosorbent assay (ELISA). We now describe an immunoblot test for antibodies to T. ovis based on the recognition of a 94-kDa antigen band in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile of T. ovis scolex secretions. The test was specific for cestode infections in dogs and was 82% sensitive for the recognition of T. ovis infections. Affinity-purified antibody uniquely recognizing this 94-kDa band was used to screen a cDNA expression library constructed utilizing mRNA from newly evaginated T. ovis scoleces. The cDNA from putative positive bacteriophage were subcloned into the plasmid pGEX and expressed as fusion proteins with schistosome glutathione-S-transferase (GST). The expressed fusion proteins were purified using glutathione-agarose beads. The recombinant parasite antigen was either eluted as a fusion protein with GST or cleaved from GST using a restriction protease. Some dog sera reacted with the GST molecule. However, the recombinant cleaved antigen from 1 clone, T. ovis 40, showed 42% sensitivity and 100% specificity for cestodes in an ELISA using test sera from dogs monospecifically infected with T. ovis and preabsorbed with bacteria. Some sera from dogs monospecifically infected with other cestode species (Taenia pisiformis, 30%; Taenia hydatigena, 30%; Echinococcus granulosus, 20%) reacted with the cloned antigen.

Animals↗

PubMed1994

Cross-resistance between cisplatin and antimony in a human ovarian carcinoma cell line.

The metal compound cisplatin (DDP) is a widely used anticancer agent but naturally occurring and acquired resistance to DDP limits its effectiveness. Resistance is associated with a decreased accumulation of DDP and increased levels of glutathione and metallothioneins. Such changes also serve as protective and detoxification mechanisms for other metal salts in prokaryotes and lower eukaryotes. The aim of this study was to find metal salts for which the cross-resistance profile was the same as for DDP in sublines of the parental 2008 human ovarian carcinoma cells selected with either DDP (2008/C13*5.25) or CdCl2 and ZnCl2 (2008/MT). Among the metal salts tested the resistance profile of trivalent antimony most closely resembled that of DDP. DDP-selected cells were 15-fold resistant to DDP and 4.4-fold cross-resistant to antimony potassium tartrate, whereas of the cations tested (Cd2+, Zn2+, Ni2+ and Co2+) cross-resistance was observed only for Cd2+ (2.4-fold). When 2008 cells were selected for resistance to antimony (6.6-fold) they were found to be 16-fold cross-resistant to DDP. Accumulation of the DDP analogue cis-[3H]dichloro(ethylenediamine)platinum(II) was 59% lower in the DDP-selected subline and 48% lower in the antimony-selected variant than in the parental cell line. We conclude from the mutual cross-resistance to DDP and antimony potassium tartrate and from the impaired uptake of [3H]DEP in both the DDP and antimony-selected variants that DDP and antimony share a common mechanism of resistance. The significance of this observation lies in the fact that several evolutionarily conserved mechanisms for antimony detoxification are already known in lower organisms which may point the way to identification of additional DDP resistance mechanisms in mammalian cells.

Antimony↗

PubMed1994

Ultrastructure of oncosphere and early stages of metacestode development of Echinococcus granulosus.

The ultrastructure of the oncospheres and developing metacestodes in vitro of Echinococcus granulosus was studied with emphasis on the origin of the laminated layer. The activated oncospheres measured 28 microns (S.D. = 1.83), and by day 5 of in vitro culture the metacestodes attained diameters up to 52 microns (S.D. = 2.66). The oncospheral plasma membrane of the tegument appeared to be formed by a microtubular cytoskeleton arranged in a predetermined pattern. By day 2 there were three main types of vesicles in the perikaryon: Vd, Vg1 and Vg2. Vesicles appeared to be synthesized in the perikaryon and continuously transported to the periphery, where the Vd vesicles could contribute to the ground substance of the syncytial tegument. We suggest that the Vg1 and Vg2 vesicles contribute to the laminated layer where, by day 5, they were seen in increased density and forming aggregations on the outer border.

Animals↗

PubMed1994

Echinococcus granulosus: the mechanism of oncosphere lysis by sheep complement and antibody.

A heat-labile component of normal sheep serum (56 degrees C for 30 min but not 50 degrees C for 30 min) was able to lyse oncospheres in vitro. The degree of effect, and the proportion of oncospheres lysed, was related to the concentration of normal unheated sheep serum complement, or other sources of complement (rabbit, mouse) in the culture. Lower concentrations were required for lysis if the culture serum was obtained from sheep immune to E. granulosus infection. Heat inactivation of normal or immune sheep serum removed any lytic ability. No lysis occurred in any concentration of unheated foetal lamb serum. However, unheated foetal lamb serum was able to restore the lytic effect to heated normal or immune serum. This suggests that lysis in both immune and normal serum is antibody-dependent and complement-mediated. The lysis in normal serum would appear to be due to natural cross-reacting antibodies that can fix complement at the oncosphere surface. The complement lesion resulted in damage to the plasma membrane. This then peeled back, predisposing the oncosphere to osmotic destruction. The use of bleach to dissolve the embryophore caused damage to the plasma membrane similar to that caused by complement. Developing metacestodes at 3 days of age in vitro in immune serum were susceptible to the addition of complement at that time.

Animals↗

PubMed1994

DNA fingerprinting used to test for family effects on precocious sexual maturation in two populations of Oncorhynchus tshawytscha (Chinook salmon).

Two single locus Variable Number Tandem Repeat (VNTR) DNA probes were used to test for differences in allele distribution between precociously mature male and immature chinook salmon, Oncorhynchus tshawytscha. Two populations were examined: Robertson Creek (RC) adult salmon, and Nicola River (NR) freshwater juveniles, or parr. Genomic DNA was extracted from 74 RC precociously mature adult males ('jacks') and 94 RC immature adults of the same age and from 45 NR precociously mature parr and 51 NR nonmaturing parr. The genomic DNA was hybridized with a single locus VNTR probe developed for chinook salmon (OtSL1), as well as one developed for Atlantic salmon, Salmo salar (Ssa1). The allele frequency distributions at both loci were significantly different for the RC jacks and immature fish, indicating a family effect on the incidence of precocious maturation in that population. No difference was found between the allele frequency distribution of the NR precocious and immature parr. A bin width sensitivity analysis showed that the comparisons of the allele frequency distributions were insensitive to the choice of bin size. No differences in heterozygosity were found between mature and immature fish at either locus for both stocks. Preliminary testing for family effects on phenotypes of interest, such as alternative life history strategies, can be performed using hypervariable VNTR DNA probes, prior to implementing costly and involved breeding programmes.

Alleles↗