Antenatal genetic diagnosis: current status and future prospects.

The current status of antenatal genetic diagnosis is reviewed and the limitations of present techniques are discussed. It is suggested that multidisciplinary clinics are the most efficient means of providing this aspect of health care. Advances in cell culture techniques, in ultrasonography and in fetoscopy will extend the services available, and the impact of this will be felt by the community. Education of the medical profession and the public in this area is necessary so that informed decision-making can take place.     

The biogeography of AustralianDaphnia: clues of an ancient (>70 m.y.) origin for the genus

The AustralianDaphnia (D. occidentalis, D. jollyi, D. lumholtzi, D. carinata, D. nivalis, andD. cephalata) include representatives from three distinct groups within the genus. The worldwide distribution of species within these three groups is consistent with a Gondwanan or pre-Gondwanan origin for each group. These data suggest an even more ancient origin for the genusDaphnia. The finding thatD. cephalata must have been separated fromD. carinata since Gondwanan times is consistent with recent biochemical data on the phylogenetic relations of AustralianDaphnia, but contrasts with electrophoretic evidence of continuing hybridization between these two taxa. It is suggested that continued gene exchange between otherwise discrete species, or clonal complexes, ofDaphnia may be maintained for millions of years, partly as a result of the varying degrees of cyclical parthenogenesis, and partly as a result of the highly fragmented population structures, found in these taxa.     

Indo-West Pacific patterns of genetic differentiation in the high-dispersal starfish Linckia laevigata

Genetic variation in four natural populations of the starfish Linckia laevigata from the Indo-West Pacific was examined using restriction fragment analysis of a portion of the mtDNA including the control region. Digestion with seven restriction enzymes identified 47 haplotypes in a sample of 326 individuals. Samples collected from reef sites within each location were not significantly differentiated based on Φ_ST or spatial distribution of haplotypes, indicating that dispersal is high over short to moderate distances. Evidence of gene flow is further supported by the low divergence among haplotypes and the lack of any clear geographical structuring among different haplotypes in the gene phylogeny. However, analysis of molecular variance ( AMOVA ), Φ_ST and contingency χ² analyses of the spatial distribution of haplotypes demonstrate the presence of significant broad scale population genetic structure among the four widespread locations examined. RFLP data are consistent with high gene flow between the Philippines and Western Australia and moderate gene flow between the Great Barrier Reef (GBR) and Fiji, but only limited gene flow between either the Philippines or Western Australia and either the GBR or Fiji. The presence of mtDNA structure contrasts with previous allozyme data which suggest that dispersal among widely separated locations is equivalent to dispersal among populations within the highly connected GBR studies. This discordance between patterns of gene flow inferred from these two markers cannot be fully accounted for by differences in effective population size for mtDNA. This might suggest that while mtDNA variation may represent contemporary patterns of gene flow, allozyme variation among populations is yet to reach equilibrium between drift and migration over the range surveyed.     

Initiation reactions in the mRNA-dependent reticulocyte lysate

Reticulocyte lysates depleted of mRNA by digestion with micrococcal nuclease still show an unexpectedly high rate of formation of 80 S initiation complexes. Formation of these complexes is sensitive to all inhibitors of the normal protein synthesis initiation process tested. Such lysates contain high concentrations of mRNA fragments which can be utilized for initiation, with which exogenous mRNA must compete. As a consequence of this competition, mRNAs that are weak initiators may be translated poorly by this system even at low exogenous mRNA concentrations.     

Simultaneous automated measurement of total ‘antioxidant’ (reducing) capacity and ascorbic acid concentration

Summary

Biochemical tests of antioxidant status may be useful in measuring oxidative stress, however the time, skills and resources required for most such tests have limited their application and evaluation. In this study, newly available tests for two indices of antioxidant status, ferric reducing ability of plasma (FRAP) and ascorbic acid, were combined in one simple, rapid, automated test, referred to as FRASC.

Results are available within 5 min of sample/reagent mixing. Linear dose-response is seen over a wide concentration range for both parameters. In- and between- run CVs were <1.0% and <3.0% respectively at 900–1800 μmol/l FRAP, and <5.0% for both at 20–400 μmol/l ascorbic acid. Mean (SD) FRAP values and ascorbic acid concentrations of fasting plasma from 130 healthy adults were 1018 (198) and 51 (18) μmol/l respectively.

This combined test is straightforward and inexpensive, and lends itself to the measurement of ‘antioxidant’ (reducing) capacity and ascorbic acid concentrations of various biological fluids, as well as extracts of drugs and foodstuffs. FRASC will, therefore, facilitate experimental and clinical studies of pro-oxidant: antioxidant balance and oxidative stress-related disease.     

Assessment of membrane protection by traditional Chinese medicines using a flow cytometric technique: preliminary findings

In this preliminary study, we used a 'living cell' flow-cytometric approach to membrane protection by four traditional Chinese medicines (TCMs). Cells were incubated, separately, for 30 min with aqueous extracts (1.5% w/v) of lingzhi (Ganoderma lucidum), ginger (Zingiber officianale), ginseng (Panax ginseng), and green tea (Camellia sinensis). Membranes were labelled with a fluorescent probe, cells were then incubated with cumene hydroperoxide, and site-specific oxidation induced by iron/ascorbate. Oxidation of membrane lipids quenches fluorescence. Forward-scatter fluorescence was measured at timed intervals after initiation of oxidation. Results indicate that lingzhi and ginger contain antioxidant component(s) that act within the cell membrane and slow lipid peroxidation in situ. Results demonstrate also that this living cell model is a useful biomonitoring tool to help determine molecular aspects of putative health effects of TCMs.     

The complete sequence of the mitochondrial genome of the crustacean Penaeus monodon: are malacostracan crustaceans more closely related to insects than to branchiopods?

The complete sequence of the mitochondrial genome of the giant tiger prawn, Penaeus monodon (Arthropoda, Crustacea, Malacostraca), is presented. The gene content and gene order are identical to those observed in Drosophila yakuba. The overall AT composition is lower than that observed in the known insect mitochondrial genomes, but higher than that observed in the other two crustaceans for which complete mitochondrial sequence is available. Analysis of the effect of nucleotide bias on codon composition across the Arthropoda reveals a trend with the crustaceans represented showing the lowest proportion of AT-rich codons in mitochondrial protein genes. Phylogenetic analysis among arthropods using concatenated protein-coding sequences provides further support for the possibility that Crustacea are paraphyletic. Furthermore, in contrast to data from the nuclear gene EF1alpha, the first complete sequence of a malacostracan mitochondrial genome supports the possibility that Malacostraca are more closely related to Insecta than to Branchiopoda.     

GENETIC STRUCTURE OF GIANT CLAM (TRIDACNA MAXIMA) POPULATIONS IN THE WEST PACIFIC IS NOT CONSISTENT WITH DISPERSAL BY PRESENT-DAY OCEAN CURRENTS

The Pacific marine biota, particularly species with long planktonic larval stages, are thought to disperse widely throughout the Pacific via ocean currents. The little genetic data available to date has supported this view in that little or no significant regional differentiation of populations has been found over large geographical distances. However, recent data from giant clams has demonstrated not only significant regional differentiation of populations, but routes of gene flow that run perpendicular to the main present-day ocean currents. Extensive surveys of genetic variation at eight polymorphic loci in 19 populations of the giant clam Tridacna maxima, sampled throughout the West and Central Pacific, confirmed that the patterns of variation seen so far in T. gigas were not unique to that species, and may reflect a fundamental genetic structuring of shallow-water marine taxa. Populations of T. maxima within highly connected reef systems like the Great Barrier Reef were panmictic (average F_ST < 0.003), but highly significant genetic differences between reef groups on different archipelagos (average F_ST = 0.084) and between West and Central Pacific regions (average F_ST = 0.156) were found. Inferred gene flow was high (N_em usually > 5) between the Philippines and the Great Barrier Reef, between the Philippines and Melanesia (the Solomon Islands and Fiji), and between the Philippines and the Central Pacific island groups (Marshall Islands, Kiribati, Tuvalu and Cook Islands). Gene flow was low between these three sets of island chains (N_em < 2). These routes of gene flow are perpendicular to present-day ocean currents. It is suggested that the spatial patterns of gene frequencies reflect past episodes of dispersal at times of lower sea levels which have not been erased by subsequent dispersal by present-day circulation. The patterns are consistent with extensive dispersal of marine species in the Pacific, and with traditional views of dispersal from the Indo-Malay region. However, they demonstrate that dispersal along present-day ocean surface currents cannot be assumed, that other mechanisms may operate today or that major dispersal events are intermittent (perhaps separated by several thousands of years), and that the nature and timing of dispersal of Pacific marine species is more complex than has been thought.