Presymptomatic testing for Huntington's disease

Summary Presymptomatic testing for Huntington's disease (HD) is possible through the use of restriction fragment length polymorphisms (RFLPs) at the closely linked D4S10 locus. Recombination between the HD and D4S10 loci will occur in 4%–5% of meioses, and is a well-recognised complication of predictive testing. Recombination between RFLPs within the D4S10 locus is a rare event and can usually be ignored. We report a case where such an intra-locus recombination frustrated attempts to predict the chance of a high-risk individual inheriting the HD gene.     

The cyclin-dependent kinase inhibitor p21 (WAF1) is required for survival of differentiating neuroblastoma cells.

We are employing recent advances in the understanding of the cell cycle to study the inverse relationship between proliferation and neuronal differentiation. Nerve growth factor and aphidicolin, an inhibitor of DNA polymerases, synergistically induce neuronal differentiation of SH-SY5Y neuroblastoma cells and the expression of p21WAF1, an inhibitor of cyclin-dependent kinases. The differentiated cells continue to express p21WAF1, even after removal of aphidicolin from the culture medium. The p21WAF1 protein coimmunoprecipitates with cyclin E and inhibits cyclin E-associated protein kinase activity. Each of three antisense oligonucleotides complementary to p21WAF1 mRNA partially blocks expression of p21WAF1 and promotes programmed cell death. These data indicate that p21WAF1 expression is required for survival of these differentiating neuroblastoma cells. Thus, the problem of neuronal differentiation can now be understood in the context of negative regulators of the cell cycle.     

Salicylic acid-independent plant defence pathways

Salicylic acid is an important signalling molecule involved in both locally and systemically induced disease resistance responses. Recent advances in our understanding of plant defence signalling have revealed that plants employ a network of signal transduction pathways, some of which are independent of salicylic acid. Evidence is emerging that jasmonic acid and ethylene play key roles in these salicylic acid-independent pathways. Cross-talk between the salicylic acid-dependent and the salicylic acid-independent pathways provides great regulatory potential for activating multiple resistance mechanisms in varying combinations.     

NO vascular control in Duchenne muscular dystrophy

A new investigation into Duchenne muscular dystrophy (DMD) pathogenesis suggests that at least part of the muscle degeneration observed in DMD patients may result from the reduced production of muscle membrane-associated neuronal nitric oxide synthase. This reduction may lead to impaired regulation of the vasoconstrictor response and eventual muscle damage.     

Biosynthesis of dystroglycan: processing of a precursor propeptide

Dystroglycan is a cytoskeleton-linked extracellular matrix receptor expressed in many cell types. Dystroglycan is composed of alpha- and beta-subunits which are encoded by a single mRNA. Using a heterologous mammalian expression system, we provide the first biochemical evidence of the alpha/beta-dystroglycan precursor propeptide prior to enzymatic cleavage. This 160 kDa dystroglycan propeptide is processed into alpha- and beta-dystroglycan (120 kDa and 43 kDa, respectively). We also demonstrate that the precursor propeptide is glycosylated and that blockade of asparagine-linked (N-linked) glycosylation did not prevent the cleavage of the dystroglycan precursor peptide. However, inhibition of N-linked glycosylation results in aberrant trafficking of the alpha- and beta-dystroglycan subunits to the plasma membrane. Thus, dystroglycan is synthesized as a precursor propeptide that is post-translationally cleaved and differentially glycosylated to yield alpha- and beta-dystroglycan.     

Molecular systematics of Mesocestoides sPP (cestoda: mesocestoididae) from domestic dogs (Canis familiaris) and coyotes (Canis latrans)

The genus Mesocestoides Vaillant, 1863 includes tapeworms of uncertain phylogenetic affinities and with poorly defined life histories. We previously documented 11 cases of peritoneal cestodiasis in dogs (Canis familiaris L.) in western North America caused by metacestodes of Mesocestoides spp. In the current study, DNA sequences were obtained from metacestodes collected from these dogs (n = 10), as well as proglottids from dogs (n = 3) and coyotes (Canis latrans Say, 1823 [n = 2]), and tetrathyridia representing laboratory isolates of M. corti (n = 3), and these data were analyzed phylogenetically. Two nuclear genetic markers, 18S ribosomal DNA and the second internal-transcribed spacer (ITS 2), were sequenced. Phylogenetic analysis of the 18S rDNA data recovered a monophyletic group composed of all samples of Mesocestoides spp., distinct from closely related outgroup taxa (Amurotaenia Akhmerov, 1941 and Tetrabothrius Rudolphi, 1819). Initial analysis of the ITS 2 data resolved 3 clades within Mesocestoides. Two proglottids from dogs formed a basal clade, a second clade was represented by tetrathyridial isolates, and a third clade included all other samples. Interpretation of these data from an apomorphy-based perspective identified 6 evolutionary lineages. We also assessed whether metacestodes from dogs (n = 4) are capable of asexual proliferation in laboratory mice. One tetrathyridial and 2 acephalic isolates from dogs proliferated asexually. Further investigation is warranted to determine which of the lineages represent distinct species and to determine the life history strategies of Mesocestoides spp.     

Long-term response of the biotic community to fluctuating water levels and changes in water quality in Cootes Paradise Marsh,a degraded coastal wetland of Lake Ontario

During the early 1900s, more than 90% of the surface area of Cootes Paradise Marsh was covered with emergent vegetation; currently, less than 15% of the surface is covered with aquatic vegetation and the remainder is wind-swept, turbid, open water. The loss of emergent cover is significantly correlated with mean annual water levels that increased more than 1.5 m over the past 60 years. Species diversity and the percent cover of the submerged macrophtye community also declined dramatically after the 1940s, coincident with decreased water clarity and increased nutrients from pollution by sewage and stormwater effluent. Phosphorus levels in the marsh dropped ten-fold after the sewage plant was upgraded to a tertiary-treatment facility in 1978; however, there was no measurable improvement in water clarity, in spite of a decrease in chlorophyll concentrations. Long-term changes in the composition of the planktonic, benthic and fish communities accompanied changes in water clarity, nutrient status and macrophyte cover. Phytoplankton changed from a community dominated by diverse taxa of green algae and diatoms during the 1940s, to a less diverse community dominated by a few taxa of green and blue-green algae in the 1970s, then to a much more diverse community recently, including many taxa of green algae, diatoms and chrysophytes; however, because water turbidity continues to be high, and algae tolerant of low light levels are now very abundant. Daphnia, which were prominent during the 1940s (especially in the vegetated sites) were replaced in the 1970s by smaller zooplankton such as the cladoceran, Bosmina, and several rotifer species including Brachionus, Asplanchna and Keratella. In the recent survey conducted in 1993 and 1994, small-bodied forms still dominate the turbid open-water areas, while medium-sized cladocerans such as Moina were common near macrophyte beds. Generally, total herbivorous zooplankton biomass tended to be highest next to Typha beds and declined with increasing distance from the plants. Conversely, biomass of edible algae at these sites increased with distance from the macrophytes. Species diversity of aquatic insects declined dramatically over the past 40 years, from 57 genera (23 families and 6 orders) in 1948, to 9 genera (6 families and 3 orders) in 1978, to only 5 genera (3 families and 2 orders) in 1995. The diverse benthic community present 5 decades ago has now been replaced by a community consisting primarily of chironomid larvae, oligochaetes and other worms associated with low-oxygen environments. These successional changes illustrate the impact of natural (fluctuating water levels) and anthropogenic (deterioration in water quality) stressors on the character of the biotic communities, and reveal the complex interactions among the various trophic levels and the abiotic environment as degradation and remediation proceeded.