STUDIES ON THE MECHANISM OF DEMYELINATION: MYELIN AUTOLYSIS IN NORMAL AND EDEMATOUS CNS TISSUE

The effects on myelin of autolysis in situ after death and after purification were studied in normal brains and spinal cords and in those made edematous as a result of chronic triethyl tin (TET) feeding. Myelin prepared from normal and edematous brains and spinal cords autolyzed for 12 h at 4°C contained only slightly less basic protein than that prepared from freshly killed animals. The amounts of a light lipid-protein fraction (dissociated myelin) usually obtained during purification of myelin from edematous CNS were about the same in tissue from freshly killed rats and those autolyzed for 12 h at 4°C. Autolysis for 12 h at room temperature resulted in formation of large amounts of dissociated myelin and loss of basic protein, but more dissociation and basic protein loss occurred in CNS from edematous brains and spinal cords than from the normal. Purified myelin prepared from freshly-killed normal and TET-fed rats was incubated at 37°C in media of several ionic strengths. In Krebs-Ringer bicarbonate (physiological extracellular fluid) extensive dissociation of myelin occurred with much less in 0.04 M-Tris buffer, pH 7.2, and only small amounts were formed in 0.01 M-Tris. In all cases myelin from edematous CNS formed more dissociated fraction than did the normal myelin. Basic protein loss was also proportional to the ionic strength of the media, but there was no difference in loss between normal and TET-myelin. Two different factors, proteolysis and physical extraction of basic protein by salt solutions, may be contributing to myelin dissociation and loss of basic protein.     

The use of steam/air mixtures for partially sterilizing soils infested with cucumber root rot pathogens

An apparatus for experimentally heating glasshouse soils in situ with steam/ air mixtures is described. Covering soil with a rigid hood enabled mixtures to be introduced at ground level at pressures sufficient to give reasonable rates of heat penetration. Heat-treating soils (66–100° C), in which cucumbers had previously grown poorly, increased yields of subsequent crops from (1) 33 to 39/43 kg/plot of four plants and (2) 95 to 178/191 kg/plot. These increases were associated with the eradication of a dark sclerotial pathogenic fungus causing tap root rot. Increasing partial sterilization temperatures increased concentrations of soluble and exchangeable soil Mn from 4 p.p.m. in unheated soil to 9, 43, 87 and 108 p.p.m. at 66, 77, 88 and 100° C respectively, these differences being paralleled by concentrations of Mn in leaf laminae. Cucumbers grown in soils sterilized at 88° C yielded 8 % more than those in soils treated at 100° C.     

Simultaneous Expression of Two Different Types of Intermediate Sized Filaments in Mouse Keratinocytes Proliferating in vitro

The intermediate-sized filaments present in epidermal keratinocytes derived from mouse skin and in an established cell line (HEL) derived from spontaneous transformation of murine keratinocytes grown in vitro, have been examined by immunofluorescence microscopy, using antibodies directed against subunit proteins of different classes of intermediate-sized filaments, as well as by electron microscopy and gel electrophoresis of cytoskeletal preparations highly enriched in intermediate-sized filaments. The keratinocytes derived from neonatal skin, which are capable of only limited replication in vitro, show only a single type of intermediate-sized filaments, i.e., the tonofibril-like arrays of filaments containing prekeratin. HEL cells, which proliferate indefinitely in vitro, retain the tonofilament-like structures typical of differentiated epidermal cells but in addition display intermediate-sized filaments of the vimentin type, i.e., the filament system typically found in mesenchymal and mesenchyme-derived cells. We discuss the possibility that (i) the advent of vimentin-type filaments in epidermal cells in culture is related either to the transformed state or the in vitro growth conditions as such and (ii) other differentiated epithelial cells proliferating in vitro may have more than one system of intermediate-sized filaments.     

THE MANNER OF TRANSMISSION OF SOME BARLEY YELLOW-DWARF VIRUSES BY DIFFERENT APHID SPECIES

Some barley yellow-dwarf (BYD) viruses isolated from cereal crops in Great Britain were transmitted by Rhopalosiphum padi , L. and others were not. Sitobion fragariae (Walker), S. avenae (Fabricius), and Metopolophium dirhodum (Walker) all transmitted viruses of both types, but they usually transmitted those of which Rhopalosiphum was a vector less readily than did R. padi. The transmissibility of a virus by a given aphid species was not affected by transmission with another, less efficient, vector species. Neomyzus circumflexus (Buckt.) and Rhopalosiphum maidis (Fitch) transmitted the few viruses with which they were tested.
A few R. padi acquired virus from infected leaves during 30 min. feeding and inoculated healthy seedlings during 15 min. feeding, but the minimum total time taken to acquire and transmit was 10 hr. and 32 hr. were needed for about half the aphids that were able to acquire and transmit virus to do so. This may indicate the existence of a short latent period of the virus in the vector, although the evidence is not conclusive. The times spent on infected plants influenced the results more than those spent on healthy ones; many transmissions occurred with short feeding times on healthy plants so long as the time spent on infected leaves was long, but the reverse was not true. Nymphs of R. padi that moulted after they left infected plants on which they fed long enough to become infective, infected slightly fewer plants than adults fed for the same times.     

Degradation of native and exotic riparian plant leaf litter in a floodplain pond

1. A litter‐bag experiment was undertaken in a pond on the margins of a large temperate floodplain in south‐western France to assess the potential influence of the replacement of native by exotic riparian species on organic matter degradation. We determined initial litter chemical composition, breakdown rates and the invertebrate assemblages associated with the litter for five pairs of native dominant and exotic invasive species co‐occurring at different stages along a successional gradient. 2. Litter chemical composition, breakdown rates and abundance and diversity of detritivorous invertebrates were similar for the exotic and native species overall. No overall changes in organic matter degradation can thus be predicted from the replacement of dominant natives by exotic invasives. Breakdown rates were primarily driven by the C/N ratio. 3. One invasive species (Buddleja davidii) showed significantly higher breakdown rates than its native counterpart (Populus nigra), resulting in the disappearance of leaf litter 6 months prior to the next litterfall. In some cases, therefore, invasion by exotic species may result in discontinuity of resource supply for decomposers.     

Inhibition of the development of the reproductive tract in parasitized snails

Summary

Myoblasts, muscle cells with the capacity to divide, have been detected in “Anlagen” of the male copulation organ of Lymnaea stagnalis. They only occur in the apical part of the penis. Here they could be found throughout life. Mitotic activity of these cells can be demonstrated by using an antiserum to a S-phase specific cell cycle marker, PCNA [see, e.g., Baserga (1991)]. The number/percentage of PCNA positive myoblasts is a good parameter for growth of this male copulation organ and hence also for inhibition of its growth and development as occurs in parasitized snails. In transplantation experiments, “Anlagen” of the copulation organ were used from snails 7–9 weeks after being parasitized as they can be excised in this stage and transplanted into either parasitized or nonparasitized snails. These experiments have indicated that humoral, parasitic excretory/secretory factors can be responsible for the inhibition of growth and differentiation of the copulation organ in parasitized snails as reflected by a relatively low number of PCNA positive myoblasts compared to the controls. Data obtained in in vitro experiments showed a significant decrease of the number of myoblasts in “Anlagen” cultured in the presence of parasitic E/S products. The fact that no significant effect was found on the relative low number of PCNA positive myoblasts is discussed. The effect of parasitic E/S products on these myoblasts appeared to be exerted in a direct way, not mediated by CNS-derived factors or by factors from cells in the connective tissue sheath around the CNS. Although it appears possible to use transplantation and/or in vitro culturing of these “Anlagen” as a bioassay for identification of the parasitic factor(s) responsible for the inhibitory effects on myoblasts, the methods are very laborious and do not seem very appropriate for testing many fractions of E/S products.     

Cellular and Humoral Immunity: a Distinction in Antigenic Recognition

THERE is little or no humoral cross reactivity between the native and denatured forms of a protein antigen. Antibodies to native bovine ribonuclease do not bind ribonuclease oxidized with performic acid^1–3, indicating that antibodies can distinguish between two molecular conformations. There is no humoral cross reactivity between rabbit IgG and the product of reduction of all the disulphide bridges^4–6. Also there is essentially no serological cross reactivity between lysozyme and its reduced and S-carboxymethylated (CM-lysozyme) derivative^7–9.     

Different pathotypes of the sunflower downy mildew pathogen Plasmopara halstedii all contain isometric virions†

Eight pathotypes of Plasmopara halstedii were screened to investigate the occurrence of virions and the potential viral influence on the pathogenicity of the sunflower downy mildew pathogen. In 23 of 26 P. halstedii isolates derived from eight countries in Europe, North America and South America, virions were detected by transmission electron microscopy. By contrast, there were no ultrastructural indications of virus‐like particles in eight other related Oomycetes. The virions of representative P. halstedii isolates were morphologically and biochemically characterized and compared among each other. Regardless of their host's pathotypes, the geographical origin of the isolate and the sensitivity towards the fungicide metalaxyl, the viral characters obtained were uniform. The virions were isometric and measured approximately 37 nm in diameter. One polypeptide of c. 36 kDa and two segments of single‐stranded RNA (3.0 and 1.6 kb) were detected. Both viral RNA segments were detected by capillary electrophoresis in the three remaining P. halstedii isolates where virions were undetectable by transmission electron microscopy. Virus‐specific primers for the 1.6 kb‐segment were synthesized and used to determine and compare a partial sequence of the viral coat protein among virions of different P. halstedii pathotypes. In all tested isolates, fragments of 0.7 kb were amplified which were directly sequenced. Sequence variation was insignificant. As both less aggressive and more aggressive P. halstedii isolates contained virions, the presence or absence of virions could not explain the diverse aggressiveness of the downy mildew pathogen towards sunflower. Moreover, the results indicated that pathogenicity of P. halstedii was not related to variation in morphological or biochemical characters of the virions.