Haplotype mining panel for genetic dissection and breeding in Eucalyptus

To improve our understanding of genetic mechanisms underlying complex traits in plants, a comprehensive analysis of gene variants is required. Eucalyptus is an important forest plantation genus that is highly outbred. Trait dissection and molecular breeding in eucalypts currently relies on biallelic single-nucleotide polymorphism (SNP) markers. These markers fail to capture the large amount of haplotype diversity in these species, and thus multi-allelic markers are required. We aimed to develop a gene-based haplotype mining panel for Eucalyptus species. We generated 17 999 oligonucleotide probe sets for targeted sequencing of selected regions of 6293 genes implicated in growth and wood properties, pest and disease resistance, and abiotic stress responses. We identified and phased 195 834 SNPs using a read-based phasing approach to reveal SNP-based haplotypes. A total of 8915 target regions (at 4637 gene loci) passed tests for Mendelian inheritance. We evaluated the haplotype panel in four Eucalyptus species (E. grandis, E. urophylla, E. dunnii and E. nitens) to determine its ability to capture diversity across eucalypt species. This revealed an average of 3.13–4.52 haplotypes per target region in each species, and 33.36% of the identified haplotypes were shared by at least two species. This haplotype mining panel will enable the analysis of haplotype diversity within and between species, and provide multi-allelic markers that can be used for genome-wide association studies and gene-based breeding approaches.     

EATB Donor Case Workshop 2007

The European Association of Tissue Banks (EATB) Donor Case Workshop is a forum held within the programme of the EATB annual Congress since 2003. This workshop has been used to discuss clinical donor cases with peer review of practice. It was agreed in advance that the experience of the 2007 workshop should be shared by publication as an example of participative learning which can be extended to other fields within tissue banking and which may be applicable in other disciplines. The EATB Congress in 2008 will extend the idea of participative open workshops with two additional workshops, one on Quality System cases and another on heart valve cases.     

Possible role of acrolein in oxazaphosphorine-induced enhancement of immunological reactivity

Summary The aim of the present study was to analyze further the immunopotentiating effects of low doses of oxazaphosphorines. We examined 4-hydroperoxycyclophosphamide (4-HC) and mafosfamide, which degrade spontaneously in water without requiring liver enzymes to become active. Both drugs, at concentrations ranging from 0.01 µM to 1 µM, enhanced mitogenic responses of human lymphocytes. Higher concentrations were toxic. Acrolein, which is one of the degradation products of oxazaphosphorines, had similar effects. Immunopotentiation was not monocyte-dependent. Attempts to inactivate released acrolein with human serum reduced toxicity but the immunostimulating property of the drugs remained Similar effects were noted when lymphocytes were exposed to acrolein dissolved in serum. 2-Mercaptoethanesulfonate (mesna), which is highly reactive with acrolein, reduced the toxicity of solutions of both oxazaphosphorines and acrolein. Immunopotentiation was not clearly demonstrable since mesna itself enhanced the responses. Pretreatment of lymphocytes with 4-HC or mafosfamide did not reduce the capacity of concanavalin A to induce suppressor cells. It is speculated that acrolein may play a role in oxazaphosphorine-induced enhancements of immune responses.