Rational design of point mutation-selective antisense DNA targeted to codon 12 of Ha-ras mRNA in human cells.

Antisense oligodeoxynucleotides targeted to Ha-ras mRNA have been designed to discriminate between the codon 12-mutated oncogene and the normal proto-oncogene. An in vitro assay using two different sources of RNase H (rabbit reticulocyte lysates and nuclear extract from HeLa cells) was used to characterize oligonucleotide binding to normal and mutated Ha-ras mRNA. Short oligonucleotides (12- or 13mers) centered on the mutation had a very high discriminatory efficiency. Longer oligonucleotides (16mers) did not discriminate efficiently between the mutated and the normal mRNA. We have tested the efficacy of dodecanucleotides to induce RNase H cleavage of the full-length mRNA, moving the target sequence from the loop to the stem region which is formed in the vicinity of mutated codon 12. The most selective oligonucleotides were centered on the mutation which is located near the junction between the loop and stem regions even though they were less efficient at inducing RNase H cleavage than those targeted to the loop region. The 12mer antisense oligonucleotide with the highest discriminatory power was selected for cell culture studies. This oligonucleotide inhibited the proliferation of a human cell line which had been transformed with the mutated Ha-ras gene (HBL100ras1) but had no effect on the parental cell line which was transfected with the vector DNA (HBL 100neo) and expressed only the normal Ha-ras gene. Growth inhibition of HBL100ras1 cells was associated with specific ablation of targeted Ha-ras mRNA as shown by RT-PCR. These results show that 'in vitro' evaluation using an RNase H assay allowed us to select an antisense oligonucleotide which elicited a selectivity towards point-mutated Ha-ras mRNA when added at 10 microM concentration to the culture medium of cells expressing wild type and mutated Ha-ras mRNA.     

Report of the International Equine Gene Mapping Workshop: male linkage map

The goal of the First International Equine Gene Mapping Workshop, held in 1995, was the construction of a low density, male linkage map for the horse. For this purpose, the International Horse Reference Family Panel (IHRFP) was established, consisting of 12 paternal half-sib families with 448 half-sib offspring provided by 10 laboratories. Blood samples were collected and DNA extracted in each laboratory and sent to the Lexington laboratory (KY, USA) for dispatch in aliquots to 14 typing laboratories. In total, 161 markers (144 microsatellites, seven blood groups and 10 proteins) were tested for all families for which the sire was heterozygous. Genealogies and typing data were sent for analysis to the INRA laboratory (Jouy-en-Josas, France) according to a specific format and entered into a database with input verification and output processes. Linkage analysis was performed with the CRIMAP program. Significant linkage was detected for 124 loci, of which 95 were unambiguously ordered using a multipoint analysis with an average spacing of 14.2 CM. These loci were distributed among 29 linkage groups. A more comprehensive analysis including synteny group data and FISH data suggested that 26 autosomes out of 31 are covered. The complete map spans 936 CM.     

Occupancy frequency distributions: patterns,artefacts and mechanisms

Numerous hypotheses have been proposed to explain the shape of occupancy frequency distributions (distributions of the numbers of species occupying different numbers of areas). Artefactual effects include sampling characteristics, whereas biological mechanisms include organismal, niche-based and meta-population models. To date, there has been little testing of these models. In addition, although empirically derived occupancy distributions encompass an array of taxa and spatial scales, comparisons between them are often not possible because of differences in sampling protocol and method of construction. In this paper, the effects of sampling protocol (grain, sample number, extent, sampling coverage and intensity) on the shape of occupancy distributions are examined, and approaches for minimising artefactual effects recommended. Evidence for proposed biological determinants of the shape of occupancy distributions is then examined. Good support exists for some mechanisms (habitat and environmental heterogeneity), little for others (dispersal ability), while some hypotheses remain untested (landscape productivity, position in geographic range, range size frequency distributions), or are unlikely to be useful explanations for the shape of occupancy distributions 'species specificity and adaptation to habitat, extinction-colonization dynamics). The presence of a core (class containing species with the highest occupancy) mode in occupancy distributions is most likely to be associated with larger sample units, and small homogenous sampling areas positioned well within and towards the range centers of a sufficient proportion of the species in the assemblage. Satellite (class with species with the lowest occupancy) modes are associated with sampling large, heterogeneous areas that incorporate a large proportion of the assemblage range. However, satellite modes commonly also occur in the presence of a core mode, and rare species effects are likely to contribute to the presence of a satellite mode at most sampling scales. In most proposed hypotheses, spatial scale is an important determinant of the shape of the observed occupancy distribution. Because the attributes of the mechanisms associated with these hypotheses change with spatial scale, their predictions for the shape of occupancy distributions also change. To understand occupancy distributions and the mechanisms underlying them, a synthesis of pattern documentation and model testing across scales is thus needed. The development of null models, comparisons of occupancy distributions across spatial scales and taxa, documentation of the movement of individual species between occupancy classes with changes in spatial scale, as well as further testing of biological mechanisms are all necessary for an improved understanding of the distribution of species and assemblages within their geographic ranges.     

Environmental tolerances of an invasive terrestrial amphipod, Arcitalitrus dorrieni (Hunt) in Britain

We investigated key physiological tolerances of the invasive euterrestrial talitrid amphipod (or landhopper) Arcitalitrus dorrieni; desiccation, salt, high and low temperatures. The critical relative humidity below which, A. dorrieni experiences desiccation stress is very high (95-100%), making it completely reliant on the leaflitter/soil microhabitat. It is tolerant of a wide range of (sea) salt concentrations (5-750 mOsmol l(-1)) but is extremely vulnerable below 5 mOsmol l(-1). A. dorrieni does not tolerate low temperatures with a mean lower limit of 1.4 degrees C, but with no individual surviving <0 degrees C. The range of upper thermal tolerance (30-37.3 degrees C) was similar to that found for other landhopper and beachflea species. Based on its tolerance to these environmental factors it is suggested that A. dorrieni has a limited potential to invade further into Britain, being restricted to areas with sufficiently high ion concentrations and mild winters.     

Plant genomics in Africa: present and prospects

Plants are the world’s most consumed goods. They are of high economic value and bring many health benefits. In most countries in Africa, the supply and quality of food will rise to meet the growing population’s increasing demand. Genomics and other biotechnology tools offer the opportunity to improve subsistence crops and medicinal herbs in the continent. Significant advances have been made in plant genomics, which have enhanced our knowledge of the molecular processes underlying both plant quality and yield. The sequencing of complex genomes of African plant species, facilitated by the continuously evolving next-generation sequencing technologies and advanced bioinformatics approaches, has provided new opportunities for crop improvement. This review summarizes the achievements of genome sequencing projects of endemic African plants in the last two decades. We also present perspectives and challenges for future plant genomic studies that will accelerate important plant breeding programs for African communities. These challenges include a lack of basic facilities, a lack of sequencing and bioinformatics facilities, and a lack of skills to design genomics studies. However, it is imperative to state that African countries have become key players in the plant genome revolution and genome derived-biotechnology. Therefore, African governments should invest in public plant genomics research and applications, establish bioinformatics platforms and training programs, and stimulate university and industry partnerships to fully deploy plant genomics, particularly in the fields of agriculture and medicine.     

Accounting for soil organic carbon role in land use contribution to climate change in agricultural LCA: which methods? Which impacts?

The International Journal of Life Cycle Assessment - Soil organic carbon (SOC) plays a key role in soil functioning and in greenhouse gas exchange with the atmosphere. Land use and land use changes...     

Un cytochrome de type P450 dans le spadice d'arum

A cytochrome of the P₄₅₀ type has been shown to exist in the microsomal fraction derived from Arum spadices. It appears to be present in greater quantities than the cytochrome b₅. It is transformed, partially at least, into a pigment of type P₄₅₀ under the action of salts and detergents.     

Abundance-range size relationships of breeding and wintering birds in Britain: a comparative analysis

Both breeding and wintering assemblages of birds in Britain exhibit positive interspecific relationships between population size and geographic range size, such that the average density of species is greater if they are more widely distributed Species in common to both assemblages, that is resident species, had greater population sizes, geographic range sizes, and densities in winter In contrast, whilst winter migrants had higher abundances than summer migrants, the range sizes of the former were disproportionately larger still, resulting in a lower density for species that only winter in Britain than for those that only breed Such differences aside, the overall form of the abundance-range size relationship is remarkably similar between the two assemblages and their constituent subsets of species     

Reproductive consequences of natal dispersal in a highly philopatric seabird

Natal and breeding dispersal have a major impact on gene flowand population structure. We examined the consequences of nataldispersal on the reproductive success (proportion of pairs rearingchicks) of colonial-breeding Thick-billed murres (Uria lomvia).Reproductive success increased with distance dispersed for thefirst and second breeding attempt. The increase in breedingsuccess leveled off at natal dispersal distances above 7 m.Our results were consistent with the idea that the relationshipbetween dispersal and reproductive success is caused by siteavailability and mate choice as birds willing to disperse fartherhad a greater choice of potential sites and mates. This hypothesiswas supported by the fact that birds dispersing farther weremore likely to pair with an experienced breeder, which increasesthe likelihood of breeding success for young breeders. Explanationsfor increasing breeding success with increased dispersal basedon inbreeding effects were unlikely because most breeding failureswere caused by egg loss rather than infertility or nestlingdeath. However, we could not explain why >50% of birds returnwithin 3 m of the natal site, despite having an up to 50% lowerreproductive success than birds dispersing 7 m or more.