A method for studying histochemical reactions in intact human dentine with a polarizing incident light microscope

Localization and distribution of non-specific esterases has been studied in intact human dentine, by reflected light microscopy. The method of specimen preparation described here permits the visualization of optical sections in depth within the specimen at high optical resolution. Non-specific esterase was found deposited as discrete bands across the tubules. or as droplets, or as a diffuse microsomal variety in the dentinal tubules and in the interglobular spaces. It was possible to distinguish the droplet variety from the microsomal variety, of esterase within the same tubule, by means of a novel optical method using antiflex and differential interference contrast systems of reflected light microscopy. It was found that the coefficient of reflection of dentine diminished gradually from the enamel to the pre-dentine and was inversely related to the scattering of light in dentine. This scattering plays an important role in the formation of the image with reflected light microscopy. The reflected light microscope offers an economically attractive alternative or a supplementary mode of microscopy to the confocal scanning microscopes for studying intact dentine at varying depths.     

Coordinating conservation: global networking for species survival

The Species Survival Commission (SSC) of IUCN—the World Conservation Union is the largest, most comprehensive and diverse professional conservation network in existence today. It brings together over 6000 volunteers from more than 160 countries to contribute to the conservation of biological diversity by developing and executing programmes to study, save, restore, and manage wisely species and their habitats. The SSC is organized primarily along taxonomic lines, with members distributed in 96 Specialist Groups focusing on distinct groups of species. These groups are a source of the most current and expert information on the conservation status and priorities for action for species in the wild. As such, they provide an invaluable resource in setting priorities for captive propagation and in linking ex situ actions. In addition, the SSC has five disciplinary Specialist Groups that provide expertise to both the zoo community and field conservationists in areas such as reintroductions, veterinary medicine, captive breeding as a conservation tool, and the impact of invasive species on native flora and fauna. The effectiveness of the SSC is greatly enhanced by the close connections within its network between zoo professionals, academic scientists, field conservationists, and managers of natural resources. Challenges to the SSC currently include better information management, organization at the national level, and extending coverage in areas such as marine biodiversity.     

Comparison of Techniques for the Production of Sclerotinia trifoliorum Ascospores in the Laboratory for Forage Legumes Resistance Tests

This study aimed to improve the production, under controlled conditions, of Sclerotinia trifoliorum ascospores to be used for selection tests on forage legumes. Sclerotia of this fungus, produced in the laboratory, were buried 1–2 cm deep in permanently soaked vermiculite, at T = 15°C, 12/24 h of fluorescent light, air moisture >80% in order to differentiate apothecia. The ascospores were harvested by aspiration and collected on a membrane filter. Various types of apparatus containing sclerotia have been tested for their ability to maintain live apothecia and for the facility of spore catching. The aspiration method has been found to be much more efficient than the previous method of cutting off apothecia. Moreover, it was observed that using this harvesting technique the best apparatus presented small and independent compartments (truncated plastic bottles). The ascospores can be stored, on the membrane filter at 5°C and in dry conditions in order to preserve their germinating abilityfor, a period of 3 months at least. This spore harvesting method permits avaibility of large quantities of a cheap inoculum for S. trifoliorum resistance test on forage legumes all year round.     

Oxidative stress responses in transgenic tobacco containing altered levels of glutathione reductase activity

A pea glutathione reductase cDNA was expressed in tobacco. Three classes of construct were used which gave a range of elevated levels of glutathione reductase (GR) activity in the cytosol (GR32), chloroplasts (GR36), or in both chloroplasts and mitochondria (GR46). In some transgenic progeny (T₂) from self-fertilized GR32 and GR36 primary transformants, having approximately twofold elevation of GR activity as compared with recessive siblings, there was an amelioration of the effect on leaf discs of up to 15 µM paraquat. However, lines with similarly elevated levels of GR activity showed no decreased sensitivity to the herbicide. None of the GR32 and GR36 lines was less sensitive to ozone. Conversely, T₂ progeny of GR46 lines, with greater than 4.5-fold elevations of GR activity, showed no reduced sensitivity to paraquat but two out of four of these lines were less sensitive to ozone fumigation. The differential response to stress co-segregated with the presence of the transgene but there was no relationship between the degree of stress response and the level of GR activity. There was an elevation in the total glutathione pool in all lines showing increased GR activity but there was no change in the ratio of oxidized to reduced glutathione. These results demonstrate that the mechanisms of protection against ozone and paraquat are different although both can be mediated by elevated GR activity.     

High Resolution Proton NMR Spectroscopy of Multiple Sclerosis Lesions

Abstract: Tissue from postmortem multiple sclerosis and normal control brains was extracted with perchloric acid and analysed using proton NMR spectroscopy. The content of N -acetyl-derived groups (the sum of N -acetylaspartate, acetate, and N -acetylaspartylglutamate) was decreased in multiple sclerosis plaques compared with normal control white matter (mean, 4.36 vs. 6.64 µmol/g wet weight). In normal appearing white matter adjacent to plaques a corresponding decrease was seen, with no change in white matter distant from plaques. A decrease in the content of total creatine was observed in multiple sclerosis plaques in comparison with normal control white matter (mean, 4.64 vs. 6.56 µmol/g wet weight), which correlated strongly with the decrease in N -acetyl-derived groups. No changes in other metabolites such as total choline or myo -inositol were seen. The decreases in content of N -acetyl-derived groups are in agreement with observations from in vivo proton NMR spectroscopy in multiple sclerosis patients. The decrease in total creatine is in contrast to most of the observations made in vivo where total creatine is assumed to be unchanged and metabolite levels are often expressed as a total creatine ratio. The use of a total creatine ratio in vivo could lead to an underestimation of reductions in N -acetylaspartate and an apparent increase in other metabolites in the multiple sclerosis lesion.     

Modulation of K+ channels by hydrogen peroxide.

External application of hydrogen peroxide (H2O2) was found to inhibit the time-dependent fast inactivation process of three cloned voltage-gated K+ channels expressed in Xenopus oocytes: KShIIIC, KShIIID and HukII. As expected from kinetic models where some channels are still opening while a significant fraction of channels is already inactivated there was a large increase in current magnitude concomitant to inactivation block. The channels otherwise functioned normally. The effects of H2O2 were specific (other cloned voltage-gated K+ channels were not affected), and reversible, the currents returned to normal upon removal of the H2O2. H2O2 is produced during normal metabolism; it could act as a modulator of excitability through effects on K+ channels if effective local concentrations are reached in neuronal regions close to the channel. KShIIIC and KShIIID currents are very similar to an O2-sensitive K+ current present in type I cells of the carotid body which is believed to underlie the modulation of excitability of these cells by changes in arterial O2 pressure. H2O2 has been proposed as an intermediary between O2 and cellular response in the carotid body; our results provide support for this model.     

The interaction of decorin core protein fragments with type I collagen.

To further define the molecular interaction between decorin and type I collagen we generated a 20 kD fragment containing the N-terminal half of the core protein by Endoproteinase Arg C digestion and a 40 kD fragment including all leucine-rich repeats in the central part of decorin core by cleavage with 2-nitro-5-thiocyanobenzoate. The fragments did not influence collagen fibril formation, even at high concentration, and radioactive fragments showed little binding to collagen fibrils. Our observations suggest that neither the N-terminal half nor the central leucine-rich repeats of the decorin core protein can, by itself, interact fully with fibrillar collagen.