Non-linearities, synergisms and plant extinctions in South African fynbos and Australian kwongan

This paper explores the determinants of extinction of endemic plant taxa in mediterranean-climate regions in South Africa and southwestern Australia. Major threats to biodiversity in these areas include agriculture, deforestation, fragmentation, invasive alien organisms and urbanization. Case studies from the two regions show that synergisms between factors can lead to discontinuous, or non-linear, responses that have increased extinction rates (or threaten to) beyond predictions based on simple deterministic processes.     

Selection for β2-microglobulin mutation in mismatch repair-defective colorectal carcinomas

Novel peptide antigens complexed with human leukocyte antigen (HLA) and β₂-microglobulin (β2M) molecules are presented at the cell surface to cytotoxic T lymphocytes (CTLs), provoking lysis of the antigen-presenting cell [1]. In tumour cells, genetically altered or abnormally expressed proteins provide a source of peptides that can be presented to CTLs; the resulting anti-tumour CTL responses may provide part of the body's defence against cancer. Disabling mutations in the HLA and β2M proteins required for peptide presentation allow a tumour cell to escape destruction by CTLs. Cells with deficient DNA mismatch repair have high spontaneous mutation rates [2] and produce many altered proteins that are a potential source of numerous unique peptides. Mutator tumour cells might therefore be particularly vulnerable to immune surveillance and CTL attack. Mutator phenotypes [3] and [4] and loss of β2M (or HLA) expression [5] and [6] are both relatively common among sporadic colorectal tumours. We have compared the frequency of β2M mutations in sporadic colorectal and other tumours with and without a mutator phenotype. Mutations were more frequent among colorectal tumours with the microsatellite instability indicative of a defect in DNA mismatch repair. The inactivating β2M mutations were predominantly frameshifts, which is consistent with the underlying mismatch repair defects. Evasion of immune surveillance by acquiring β2M mutations therefore occurs at high frequency in tumour cells with a mutator phenotype due to defective DNA mismatch repair.     

Practical Approaches to Low Density Lipoprotein Oxidation: Whys, Wherefores and Pitfalls

The purpose of this review is to bring together the different approaches for studying the oxidation of low density lipoproteins and try to identify some critical factors which will permit greater comparability between laboratories. These issues are discussed both in terms of the variety of exogenous mediators of oxidation applied (transition metal ions, haem proteins, azo initiators, peroxynitrite, cells etc.) and their raisons d'etre, as well as the methodologies (formation of conjugated dienes, hydroperoxides, decomposition products of lipid peroxidation, altered surface charge, macrophage uptake) applicable to the different stages of the oxidation and the factors underlying their accurate execution and interpretation.     

Minimizing growth regulators in shoot culture of an endangered plant,Hackelia venusta (Boraginaceae)

Summary Hackelia venusta (Boraginaceae) is an endangered perennial herb endemic to the interior northwestern United States. Because of seed scarcity, micropropagation (anex situ conservation strategy) could produce true-to-type plantlets suitable for reintroduction. We hypothesized that clones of predetermined size could be rapidly produced by supplementing multiplication and rooting media with minimal levels of cytokinin and auxin. Microshoots derived from shoot expants were cultured on Murashige and Skoog (1962) media supplemented with 1% (wt/vol) agar and 0.0001 to 10 μM benzyladenine. Inverse regression estimates on 3 genotypes predicted that a target of 2.5 axillary microshoots per explant would require a minimal level of 0.04±0.02 μM benzyladenine. Culture of 25 genotypes with 0.04 μM benzyladenine resulted in an average of 2.3±0.1 axillary microshoots per explant. Elongated microshoots were transferred to media supplemented with 0.1 to 25 μM indoleacetic acid. Clones rooted from 36% to 100% success after 4 wk in 2.0 μM indoleacetic acid. Plantlets transplantedex vitro with three or more roots survived at 84% versus 46% of plantlets with fewer roots. Up to 84% of the plantlets survived in a planting trial. The data suggest that shoot culture ofHackelia venusta, with minimal growth regulators, can produce axillary microshoots for reintroduction.     

Inclusion volume solid-phase synthesis

Solid-phase synthesis of peptides was carried out using only the volume of the solvent included in the swollen solid-phase resin beads [inclusion volume synthesis]. This approach enables (i) the use of higher concentrations of activated amino acids, resulting in increased coupling rates, (ii) drastically decreased consumption of solvents, and (iii) the construction of multiple peptide synthesizers having virtually no reaction vessels.     

Low‐dose linearity: The rule or the exception?

In 1976, Crump, Hoel, Langley, and Peto described how almost any dose‐response relationship for carcinogens becomes linear at low doses when background cancers are taken into account. This has been used, by the U.S. Environmental Protection Agency, USEPA, as partial justification for a regulatory posture that assumes low‐dose linearity, as is illustrated by a discussion of regulation of benzene as a carcinogen. The argument depends critically on the assumption that the pollutant and the background proceed by the same biological mechanism. In this paper we show that the same argument applies to noncancer end points also. We discuss the application to a number of situations: reduction in lung function and consequent increase in death rate due to (particulate) air pollution; reduction in IQ and hence (in extreme cases) mental deficiency due to radiation in utero; reduction of sperm count and hence increase in male infertility due to DBCP exposure. We conclude that, although the biological basis for the health effect response is different, in each case low‐dose linearity might arise from the same mathematical effect discussed by Crump et al. (1976). We then examine other situations and toxic end points where low‐dose linearity might apply by the same argument. We urge that biologists and chemists should concentrate efforts on comparing the biological and pharmacokinetic processes that apply to the pollutant and the background. Finally, we discuss some public policy implications of the possibility that low dose linearity may be the rule rather than the exception for environmental exposures.     

La procréation médicalement assistée avec spermatozoïdes immatures: Impératifs cliniques et techniques chirurgicales

The authors report their experience with the use of spermatids in TESE programs where mature spermatozoa could not be isolated from testicular biopsies. The details of the indications for spermatid insemination, the technicity of the procedure and the results are exposed.     

Intraspecific molecular variation inHieracium sect.Alpina (Asteraceae), an apomictic group

Intraspecific variation of four agamospecies ofHieracium sect.Alpina was studied using RAPD and isozyme techniques. No variation in either multiprimer RAPD or multi-enzyme phenotypes was observed withinH. holosericeum, suggesting that this widespread species consists of only a single genotype. A low level of within-population isozyme variation was seen inH. tenuifrons andH. calenduliflorum, the origin of which appears to be consistent with somatic mutation. Most isozyme and all RAPD variation in these two species was partitioned between populations. A strong correlation with geography suggests that its cause may be due to polytopic (-polyphyletic?) origin or perhaps to mutation and dispersal. The most variable species wasH. alpinum, in which isozyme variation occurred mostly within populations rather than between them, suggesting occasional sexual events or that the parents ofH. alpinum were heterozygous. RAPD variation in this species, in contrast, was partitioned between Scottish and Swiss populations, suggesting the existence of geographical races.     

Satellite Spitzenkörper in growing hyphal tips

Summary Growing hyphal tips of higher fungi contain an organized assemblage of secretory vesicles and other cell components collectively known as the Spitzenkörper. Until now, the Spitzenkörper has been portrayed as a single spheroid complex located near the apical cell wall. This study demonstrates the occurrence of multiple Spitzenkörper in growing hyphal apices imaged by video-enhanced phase-contrast microscopy. In addition to the main Spitzenkörper, smaller satellite Spitzenkörper arise a few micrometers behind the apical pole. Four developmental stages were identified: (a) the satellites first appeared as faint phase-dark plaques next to the plasma membrane, (b) gradually increased in size and assumed an ovoid profile, (c) they migrated to the hyphal apex, and (d) finally they merged with the main Spitzenkörper. After the merger, the main Spitzenkörper temporarily increased in size. Satellites were observed in 14 fungi, most of which had relatively large (5–10 m diam.), fast-growing hyphae (2–33 m/min elongation rate). The average frequency of in-focus satellites was 7+/min forFusarium culmorum and 11+/min forTrichoderma viride. As with the main Spitzenkörper, satellites were present only in growing cells. They were transient and remained visible for 3–8 s before merging with the main Spitzenkörper. Within the hyphae, satellites travelled up to six times faster than the average cell elongation rate. Multiple satellites sometimes occurred simultaneously; up to three were seen within a hyphal apex at the same time. Localized cell enlargement occurred next to stationary satellites, suggesting that satellite Spitzenkörper are functional as sources of new cell surface before they reach the main Spitzenkörper; therefore, they account for some variations in the profiles of the growing hyphae. By electron microscopy, satellites consisted of small clusters of apical vesicles surrounding a group of microvesicles located next to the plasma membrane. The identification and behavior of the satellites represent clear evidence of directional mass transport of vesicles toward the hyphal apex. Our observations indicate that satellites are a common phenomenon in growing hyphal apices of septate fungi and that they contribute to growth of the hyphal apex.Abbreviations VSC vesicle supply center     

Developmental regulation of aldoxime formation in seedlings and mature plants of Chinese cabbage (Brassica campestris ssp. pekinensis) and oilseed rape (Brassica napus): Glucosinolate and IAA biosynthetic enzymes

The first steps in the biosynthesis of glucosinolates and indole-3-acetic acid (IAA) in oilseed rape (Brassica napus L.) and Chinese cabbage (Brassica campestris ssp. pekinensis) involve the formation of aldoximes. In rape the formation of aldoximes from chain-extended amino acids, for aromatic and aliphatic glucosinolate biosynthesis, is catalysed by microsomal flavin-containing monooxygenases. The formation of indole-3-aldoxime from l-tryptophan, the potential precursor of both indole-3-acetic acid and indolyl-glucosinolates, is catalysed by several microsomal peroxidases. The biosynthesis of glucosinolates and indole-3-acetic acid was shown to be under developmental control in oilseed rape and Chinese cabbage. No monooxygenase activities were detected in cotyledons or old leaves of either species. The highest monooxygenase activities were found in young expanding leaves; as the leaves reached full expansion and matured the activities decreased rapidly. The indole-aldoxime-forming activity was found in all of the tissues analysed, but there was also a clear decrease in foliar activity with maturity in leaves of rape and Chinese cabbage. Partial characterisation of the Chinese cabbage monooxygenases showed that they have essentially identical properties to the previously characterised rape enzymes; they are not cytochrome P450-type enzymes, but resemble flavin-containing monooxygenases. No monooxygenase inhibitors were detected in microsomes prepared from either cotyledons or old leaves.Abbreviations DHMet dihomomethionine - FMO flavin-containing monooxygenase - HPhe homophenylalanine - IAA indole-3-acetic acid - l-Phe l-phenylalanine - l-Trp l-tryptophan - MO monooxygenase - IAALD indole-3-acetaldehyde - IAOX indole-3-aldoxime - THMet trihomomethionine