Aetiology of the skeletal dysmorphology syndrome campomelic dysplasia: expression of the Sox9 gene during chondrogenesis in mouse embryos.
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Animals↗
Biomedical subjects
J Bowles
Publications and source records attributed to J Bowles.
Molecular genetic approaches to parasite identification: their value in diagnostic parasitology and systematics.
A wide range of approaches is available to parasitologists to aid in specific parasite identification and to formulate phylogenetic relationships. This review emphasises the usefulness of molecular genetic techniques, especially DNA-based procedures, in addressing problems of identification, characterisation and phylogeny of parasites. It should be stressed that an understanding of the various DNA approaches, techniques and target genes most likely to be effective in addressing key issues in diagnostic parasitology and systematics is still developing. Nevertheless, DNA methods clearly have great potential with regard to specificity and sensitivity, and applications will increase further with technological advance. Indeed, because of the minimal requirements for material, PCR-based methods especially should prove of immense value in future studies with parasites.
Animals↗
Fresh and cryopreserved ovarian tissue samples from donors with lymphoma transmit the cancer to graft recipients.
Girls and young women who require ovariectomy or cancer therapy may consider having their own eggs, embryos or ovarian tissue stored (cryopreserved) for their own future use. Ovarian tissue is simple to collect and contains large numbers of germ cells. Transplantation of fresh and frozen-thawed ovarian tissue in healthy sheep and mice has resulted in normal live young. Similar techniques may be effective in the human but it is unclear whether ovarian tissue cryopreservation and grafting is suitable for ovaries from individuals with cancer or infections. If cancer cells were present in an ovary at the time of collection and survived cryopreservation and grafting they could establish cancer in the recipient. We therefore performed ovarian cryopreservation and transplantation trials using a mouse lymphoma model. This established that the lymphoma was transmitted by grafts of both fresh and frozen ovarian tissue. The normal healthy recipient mice died 9-43 days after receiving a small piece (1 mm3) of ovarian tissue from a donor with lymphoma. We conclude that ovarian tissue which is collected, cryopreserved and grafted while it contains cancer cells has the potential to spread the cancer to the graft recipient.
Aging↗
Scintigraphic localisation of steroid injection site in plantar fasciitis.
Plantar fasciitis is a common cause of heel pain. We evaluated scintigraphic localisation of the inflammatory focus in 15 patients with this condition. Technetium-labelled bone scans precisely localised abnormal discrete areas of tracer uptake in 12 (80%) of patients in the medial and posterior aspect below the inferior surface of the calcaneum. Steroid injection at the inflammatory site abolished local tenderness and reduced pain in all 12. Our findings support an injection approach through the medial heel border posterior to the point of heel tenderness. Technetium scintigraphy may be a useful investigation to localise the steroid injection site in resistant cases of plantar fasciitis.
Fasciitis↗
A molecular phylogeny of the human schistosomes.
Members of the genus Schistosoma are generally grouped on the basis of egg morphology, intermediate host specificity, and geographic origin. We have tested hypotheses based on these groupings by phylogenetic analysis of nuclear ribosomal (ITS2) and mitochondrial (COI) nucleotide sequences. Both mitochondrial and nuclear DNA data strongly support "traditional" hypotheses that (a) members of the Schistosoma haematobium group form a monophyletic clade, (b) members of the S. mansoni group form a monophyletic clade, (c) S. japonicum and S. mekongi form a monophyletic group relative to other schistosomes, and (d) the African schistosomes form a clade to the exclusion of the two Asian species.
Animals↗
A molecular phylogeny of the genus Echinococcus.
Three nucleotide data sets, two mitochondrial (COI and ND1) and one nuclear (ribosomal ITS1), have been investigated in order to resolve relationships among species and strains of the genus Echinococcus. The data have some unusual properties in that mitochondrial heteroplasmy was detected in one strain of E. granulosus, and more than one class of ITS1 sequence variant can occur in a single isolate. The data failed to support the hypothesis that E. granulosus, as it is currently viewed, is a single valid species. Rather, the strains of E. granulosus seem to comprise at least three evolutionarily diverse groups, the sheep strain group, bovine strain group and horse strain group. Molecular distances between them are comparable to, or greater than, molecular evolutionary distances observed between recognized species. The affinities of the cervid strain of E. granulosus are unclear because of ambiguous data, but this strain does not appear to be ancestral to others. E. multilocularis may not be distinct from E. granulosus. However, the remaining two species. E. vogeli and E. oligarthrus appear distinct and rather distant from the first two. Based on the results presented here, taxonomic revision of the genus is clearly warranted.
Animals↗
Expression of a linear Sry transcript in the mouse genital ridge.
Sry is the Y-chromosomal gene that is pivotal in the determination of sex in mammals, however the structure of the Sry transcript produced in the embryo has not been determined. We show here that the transcript expressed in the developing mouse gonad at the sex determining stage of development is linear, polyadenylated and encoded by a single exon, in contrast to the circular, apparently untranslated transcript produced in adult testes. The linear transcript was not detected in any other fetal tissue nor in any adult tissue tested, and was expressed only in the genital ridge portion of the urogenital ridge. The spatial and temporal profile of Sry expression suggests that its role in the mouse fetus is limited to initiating Sertoli cell development during testis determination.
Animals↗
A molecular genetic survey indicates the presence of a single, homogeneous strain of Echinococcus granulosus in north-western China.
A combination of DNA methods, including conventional RFLP, PCR-RFLP and mitochondrial sequence analysis did not detect genomic variation in any of 117 E. granulosus isolates examined from a range of intermediate hosts, including humans, in north-western China. The genotype infecting all hosts was indistinguishable from the widespread and well recognised sheep strain of E. granulosus. The public health implication of the study is that most, if not all, livestock infected with E. granulosus in this region will act as reservoirs of human infection.
Animals↗
Molecular genetic characterization of the cervid strain ('northern form') of Echinococcus granulosus.
We have used a number of molecular genetic approaches to characterize the cervid strain ('northern form') of Echinococcus granulosus. PCR-RFLP analysis of the nuclear ITS1 region of the rDNA repeat can readily distinguish the cervid form from other strains of E. granulosus. The complexity of the RFLP patterns obtained suggests that a number of distinct ITS1 types are present in this strain which may represent an inter-strain E. granulosus hybrid. Mitochondrial CO1 sequence of the cervid genotype was ambiguous at 18 positions and closely resembles a cluster of previously characterized E. granulosus genotypes, G1 (common, domestic sheep)/G2 (Tasmanian sheep)/G3 (buffalo). In contrast, mitochondrial ND1 sequence, although unique, suggests that the cervid form is most similar to strains represented by the G6 (camel)/G7 (pig) genotypes. We assume that the CO1 and ND1 sequences obtained for the cervid genotype are linked in a single mitochondrial genome although this is difficult to explain if conventional molecular genetics of mitochondrial DNA are assumed. Based on its unique ND1 sequence and ITS1 PCR-RFLP pattern, the cervid strain appears to represent a distinct genotype (designated G8) of E. granulosus.
Amino Acid Sequence↗
Genetic characterization of the Asian Taenia, a newly described taeniid cestode of humans.
Recent studies on the epidemiologic pattern of taeniasis in Southeast Asia have indicated the existence of a third form of human Taenia, distinguishable from Taenia saginata and T. solium. Originally termed Taiwan Taenia, and first described in Taiwanese aboriginals, this newly recognized taeniid is now generally referred to as Asian Taenia since it has since been recorded in a number of other Asian countries. Here we have used a genetic yardstick approach to determine whether the Asian Taenia should most appropriately be considered as a new, distinct species or as a subspecies, strain, or variant of T. saginata, which previous studies have shown it closely resembles. Sequence variation in the 28S rRNA and mitochondrial cytochrome c oxidase I (COI) genes of a range of taeniid cestodes and the COI and rDNA internal transcribed spacer I polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP) pattern differences in the Asian Taenia, T. saginata, and T. solium were used as markers of genetic identity. The PCR-RFLP approaches proved useful for rapid and unambiguous discrimination of Asian Taenia from the other two human species, whereas the mitochondrial and nuclear sequence comparisons indicate that the Asian Taenia is much more closely related to T. saginata than recognized taeniid species are to each other. The results support earlier conclusions that the Asian Taenia is a genetically distinct entity but is closely related to T. saginata, and suggest that its taxonomic classification as a subspecies or strain of T. saginata is more appropriate than formal designation as a new species. The very close relationship between Asian Taenia and T. saginata has public health implications in that the Asian form is unlikely to be an important cause of human cysticercosis because T. saginata cysticercosis, if it occurs at all in humans, is an extremely rare phenomenon.
Amino Acid Sequence↗
Molecular variation in Echinococcus.
In the past decade, molecular genetic approaches have revolutionised the study of population biology and taxonomy and have provided a vast accumulation of DNA sequence and other genetic data. DNA techniques, however, have only recently been applied to the study of helminth parasite populations yet, already, valuable information about population structure, genetic variation and phylogeny of the Echinococcus organisms is emerging. Some of this recently acquired molecular information is reviewed here, and its implications in terms of our understanding of variation and taxonomy within the genus Echinococcus emphasised. In particular, the current strain status of some geographically isolated populations of E. granulosus is discussed and the issue of inter- and intra-strain variability within E. granulosus examined.
Animals↗
Nuclear and mitochondrial genetic markers highly conserved between Chinese and Philippine Schistosoma japonicum.
Geographical isolates of S. japonicum, and particularly isolates from China and the Philippines, were examined at the molecular level for genetic divergence. Sequences from both nuclear and mitochondrial genomes were selected as markers of evolutionary divergence and S. mekongi and S. mansoni were included in the study for comparison purposes. Restriction fragment length polymorphism (RFLP) and PCR-RFLP analysis of the rDNA repeat unit and sequence analysis of the second internal transcribed spacer region (ITS2) within the rDNA repeat and the cytochrome c oxidase I (COI) gene of the mitochondrial genome were performed. No intra-specific variation in S. japonicum was found in the rDNA repeat and only very slight variation was detected within the COI sequence. A survey of the entire genome, using random amplified polymorphic DNA (RAPD) analysis, again showed that Chinese and Philippine S. japonicum are remarkably similar at the DNA sequence level. We were thus unable to obtain direct molecular evidence in support of previous findings, particularly those based on isoenzyme analysis, that a very high level of intra-specific variation exists in S. japonicum.
Animals↗
NADH dehydrogenase 1 gene sequences compared for species and strains of the genus Echinococcus.
Nucleotide sequences of a 471 bp region of the mitochondrial NADH dehydrogenase 1 gene were obtained for 59 Echinococcus isolates including representatives of each of the 4 recognised species. Ten distinct genotypes were detected among these isolates, including 6 within E. granulosus. This information complements and extends knowledge of inter- and intraspecific variation within Echinococcus and should prove useful in phylogenetic studies.
Animals↗
Rapid discrimination of Echinococcus species and strains using a polymerase chain reaction-based RFLP method.
Echinococcus species and genetically distinct strains of Echinococcus granulosus can be rapidly and reliably identified using a polymerase chain reaction (PCR)-linked restriction fragment length polymorphism (RFLP) method which surveys the sequence of a rapidly evolving region of the ribosomal DNA (rDNA) unit. Internal transcribed spacer 1 (ITS1) of the rDNA repeat was amplified from various isolates and the product was digested with one of a number of 4-base cutting restriction enzymes. Characteristic patterns were produced when samples within various species and strain groups were analysed. This method offers an objective, simple, highly sensitive and rapid approach for the discrimination of Echinococcus isolates and for study of other parasite complexes.
Animals↗
Molecular examination of the sympatry and distribution of sheep and camel strains of Echinococcus granulosus in Kenya.
Previous studies have shown that camel and sheep strains of the cystic hydatid parasite Echinococcus granulosus occur in Kenya. We examined 208 larval isolates and 40 worm samples of E. granulosus from various hosts in Kenya using restriction fragment length polymorphism analysis of a segment of ribosomal DNA amplified by the polymerase chain reaction. This was in an effort to determine whether additional strains of E. granulosus occur in Kenya, to examine the level of genetic heterogeneity within the sheep/dog and camel/dog strains previously identified, and to map out their intermediate host range and geographic distribution in Kenya. We confirmed the existence of the two strains in Kenya and showed that the distribution of the camel strain appears restricted to the Turkana region, where camels are kept as livestock. The intermediate host range for both strains seems to be similar except that humans appear refractory to infection with the camel strain. We have also shown that although the life-cycle patterns of the two strains overlap both geographically and in intermediate and definitive hosts, the strains maintain their homogeneous genetic identity.
Abattoirs↗
Cattle strain of Echinococcus granulosus and human infection.
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Animals↗
A genetic comparison of human and wildlife isolates of Echinococcus granulosus in Queensland: public health implications.
OBJECTIVE: To test the hypothesis that the hydatid parasite infecting macropods and dingoes in Queensland is a sylvatic strain of Echinococcus granulosus, distinct from the domestic strain which produces cysts in sheep and humans. DESIGN: Molecular biological techniques were used to compare DNA isolated from hydatid cysts from humans, local macropods and sheep from New South Wales and the United Kingdom, as well as from adult tapeworms in dingoes. SETTING: The human cysts were surgically resected from two patients seen with hydatidosis in Brisbane teaching hospitals over a one-year period. Neither patient had had previous contact with sheep farms. Macropods and dingoes were shot randomly in the localities where the patients presumably acquired their infections. Sheep liver cysts were obtained from abattoirs. METHODOLOGY: Studies comprised extraction of DNA from cysts, digestion by a series of restriction endonucleases, slab gel electrophoresis. Southern blotting and then hybridisation with defined DNA probes. Polymerase chain reaction, in combination with direct DNA sequencing, was used to compare DNA from cysts and adult worms from dingoes. RESULTS: The restriction fragment length polymorphism (RFLP) patterns of DNA from all cysts and a defined mitochondrial DNA sequence from all sources were indistinguishable. This finding is significant as both techniques can clearly distinguish between genetically distinct, well characterised strains of E. granulosus. CONCLUSIONS: Hydatid cysts are prevalent in some macropod populations and adult worms are common in dingoes. Since there are relatively few sheep-rearing areas in Queensland, contact with wild animals may be the main source of human hydatid infection in this State. The strain of E. granulosus in both patients was genetically indistinguishable from that found in macropods, dingoes and sheep from New South Wales and the United Kingdom. This strongly suggests that the domestic strain of E. granulosus, or a form very close genetically, freely infects Australian wildlife, and argues against the existence of a distinct sylvatic strain. The implications for public health are considerable.
Adult↗
Genetic variants within the genus Echinococcus identified by mitochondrial DNA sequencing.
The pattern of species and strain variation within the genus Echinococcus is complex and controversial. In an attempt to characterise objectively the various species and strains, the sequence of a region of the mitochondrial cytochrome c oxidase subunit I (CO1) gene was determined for 56 Echinococcus isolates. Eleven different genotypes were detected, including 7 within Echinococcus granulosus, and these were used to categorise the isolates. The 4 generally accepted Echinococcus species were clearly distinguishable using this approach. In addition, the consensus view of the strain pattern within E. granulosus, based on a variety of criteria of differentiation, was broadly upheld. Very little variation was detected within Echinococcus multilocularis. Remarkable intra-strain homogeneity was found at the DNA sequence level. This region of the rapidly evolving mitochondrial genome is useful as a marker of species and strain identity and as a preliminary indication of evolutionary divergence within the genus Echinococcus.
Amino Acid Sequence↗