An exceptionally high nucleotide and haplotype diversity and a signature of positive selection for the eIF4E resistance gene in barley are revealed by allele mining and phylogenetic analyses of natural populations

In barley, the eukaryotic translation initiation factor 4E (eIF4E) gene situated on chromosome 3H is recognized as an important source of resistance to the bymoviruses Barley yellow mosaic virus and Barley mild mosaic virus. In modern barley cultivars, two recessive eIF4E alleles, rym4 and rym5, confer different isolate-specific resistances. In this study, the sequence of eIF4E was analysed in 1090 barley landraces and noncurrent cultivars originating from 84 countries. An exceptionally high nucleotide diversity was evident in the coding sequence of eIF4E but not in either the adjacent MCT-1 gene or the sequence-related eIF(iso)4E gene situated on chromosome 1H. Surprisingly, all nucleotide polymorphisms detected in the coding sequence of eIF4E resulted in amino acid changes. A total of 47 eIF4E haplotypes were identified, and phylogenetic analysis using maximum likelihood provided evidence of strong positive selection acting on this barley gene. The majority of eIF4E haplotypes were found to be specific to distinct geographic regions. Furthermore, the eI4FE haplotype diversity (uh) was found to be considerably higher in East Asia, whereas SNP genotyping identified a comparatively low degree of genome-wide genetic diversity in 16 of 17 tested accessions (each carrying a different eIF4E haplotype) from this same region. In addition, selection statistic calculations using coalescent simulations showed evidence of non-neutral variation for eIF4E in several geographic regions, including East Asia, the region with a long history of the bymovirus-induced yellow mosaic disease. Together these findings suggest that eIF4E may play a role in barley adaptation to local habitats.     

DNAM-1 mediates epithelial cell-specific cytotoxicity of aberrant intraepithelial lymphocyte lines from refractory celiac disease type II patients

In refractory celiac disease (RCD), intestinal epithelial damage persists despite a gluten-free diet. Characteristic for RCD type II (RCD II) is the presence of aberrant surface TCR-CD3(-) intraepithelial lymphocytes (IELs) that can progressively replace normal IELs and eventually give rise to overt lymphoma. Therefore, RCD II is considered a malignant condition that forms an intermediate stage between celiac disease (CD) and overt lymphoma. We demonstrate in this study that surface TCR-CD3(-) IEL lines isolated from three RCD II patients preferentially lyse epithelial cell lines. FACS analysis revealed that DNAM-1 was strongly expressed on the three RCD cell lines, whereas other activating NK cell receptors were not expressed on all three RCD cell lines. Consistent with this finding, cytotoxicity of the RCD cell lines was mediated mainly by DNAM-1 with only a minor role for other activating NK cell receptors. Furthermore, enterocytes isolated from duodenal biopsies expressed DNAM-1 ligands and were lysed by the RCD cell lines ex vivo. Although DNAM-1 on CD8(+) T cells and NK cells is known to mediate lysis of tumor cells, this study provides, to our knowledge, the first evidence that (pre)malignant cells themselves can acquire the ability to lyse epithelial cells via DNAM-1. This study confirms previous work on epithelial lysis by RCD cell lines and identifies a novel mechanism that potentially contributes to the gluten-independent tissue damage in RCD II and RCD-associated lymphoma.     

In vitro and in vivo evaluation of O-alkyl derivatives of tramadol

Tramadol is a centrally acting opioid analgesic structurally related to codeine and morphine. O-Alkyl, N-desmethyl, and non-phenol containing derivatives of tramadol were synthesized to probe their effect on metabolic stability and both in vitro and in vivo potency.     

Cold hardening reduces photoinhibition of Eucalypts nitens and E. pauciflora at frost temperatures

Photoinhibition of photosynthesis at low temperatures was investigated in two species of subalpine eucalypt, Eucalypts nitens (Deane and Maiden) Maiden and E. pauciflora Sieb. ex Spreng. Imposition of an artificial cold-hardening treatment increased the frost tolerance of leaf tissue and increased tolerance to excess light. Cold-hardened seedlings of both species had a higher photosynthetic capacity than non-hardened seedlings at 6 and 16°C and lower levels of non-photochemical quenching (NPQ) at 20 and 5°C. Furthermore, hardened seedlings had faster rates of NPQ development at 5 and −3.5°C. An increase in minimal fluorescence, which indicates slowly reversible photoinhibition, was evident in all seedlings at −1.5 and −3.5°C but was less pronounced in hardened seedlings, with a threefold faster rate of development of NPQ, at −3.5°C than non-hardened seedlings. Hardened seedlings also recovered faster from photoinhibition at −3.5°C. Thus cold hardening increased tolerance to high light in these species. Differences between E. nitens and E. pauciflora in their response to excess light were small and significant only at −3.5°C. Faster recovery from photoinhibition of E. pauciflora was consistent with its occurrence in colder habitats than E. nitens. Received: 27 April 1997 / Accepted: 9 September 1997     

Development of drug resistance and the cross resistance in Shigella to certain chemotherapeutic preparations in vitro]

A total of 700 Shigella cultures isolated in different regions of the Ukraine in 1974 and occasionally in 1971-1973 were studied. No cultures resistant to furazolidone or enteroseptol were found. Under the experimental conditions resistance to nitrofuran and especially oxycholine preparations in Shigella developed slowly when the cultures were passaged in broths with increasing concentrations of the drugs. Cross resistance in Shigella was observed with respect to (a) various nitrofuran preparations, less pronounced with respect to furacryline and (b) enteroseptol and 5-NOK. No cross resistance was noted between the above groups of the chemotherapeutics and levomycetin and tetracycline. 5-NOK inhibited the resistance development to streptomycin, while enteroseptol inhibited the development of resistance to furazolidone in Shigella. The studies showed that in the near future there will be observed no mass development of resistance to furazolidone and enteroseptol in Shigella.     

Phosphate Glass Fibre Composites for Bone Repair

We investigate high-modulus degradable materials intended to replace metals in biomedical applications.These are typicallycomposites comprising a polylactide(PLA)matrix reinforced with phosphate glass fibres,which provide reinforcementsimilar to E-glass but are entirely degradable in water to produce,principally,calcium phosphate.We have made compositesusing a variety of fibre architectures,from non-woven random mats to unidirectional fibre tapes.Flexural properties in theregion of 30 GPa modulus and 350 MPa strength have been achieved-directly comparable to quoted values for human corticalbone.In collaboration with other groups we have begun to consider the development of foamed systems with structures mimickingcancellous bone and this has shown significant promise.The fibres in these foamed structures provide improved creepresistance and reinforcement of the pore walls.To date the materials have exhibited excellent cellular responses in vitro andfurther studies are due to include consideration of the surface character of the materials and the influence of this on cell interaction,both with the composites and the glass fibres themselves,which show promise as a standalone porous scaffold.