The lysosomotropic agent monodansylcadaverine also acts as a solvent polarity probe.

The autofluorescent substance monodansylcadaverine has recently been reported as a specific in vivo marker for autophagic vacuoles. However, the mechanism for this specific labeling remained unclear. Our results reveal that the common model of ion trapping in acidic compartments cannot completely account for the observed autophagic vacuole staining. Because autophagic vacuoles are characterized by myelin-like membrane inclusions, we tested whether this lipid-rich environment is responsible for the staining properties of monodansylcadaverine. In in vitro experiments using either liposomes or solvents of different polarity, monodansylcadaverine showed an increased relative fluorescence intensity in a hydrophobic environment as well as a Stokes shift dependent on the solvent polarity. To test the effect of autophagic vacuoles or autophagic vacuole lipids on monodansylcadaverine fluorescence, we isolated autophagic vacuoles and purified autophagic vacuole lipids depleted of proteins. Entire autophagic vacuoles and autophagic vacuole lipids had the same effect on monodansylcadaverine fluorescence properties, suggesting lipids as the responsible component. Our results suggest that the in vivo fluorescence properties of monodansylcadaverine do not depend exclusively on accumulation in acidic compartments by ion trapping but also on an effective interaction of this molecule with autophagic vacuole membrane lipids. (J Histochem Cytochem 48:251-258, 2000)     

M-MuLV-induced leukemogenesis: Integration and structure of recombinant proviruses in tumors

M-MuLV-specific DNA probes were used to establish the state of integration and amplification of recombinant proviral sequences in Moloney virus-induced tumors of Balb/Mo, Balb/c and 129 mice. The somatically acquired viral sequences contain both authentic M-MuLV genomes and recombinants of M-MuLV with endogenous viral sequences. All reintegrated genomes carry long terminal repeat (LTR) sequences at both termini of their genome. In the preleukemic stage a large population of cells exhibiting a random distribution of reintegrated M-MuLV genomes are seen, but during outgrowth of the tumor, selection of cells occurs leaving one or a few clonal descendants in the outgrown tumor. In this latter stage recombinant genomes can be detected. Although these recombinants constitute a heterogeneous group of proviruses, characteristic molecular markers are conserved among many individual proviral recombinants, lending credence to the notion that a certain recombinant structure is a prerequisite for the onset of neoplasia. The structure of these recombinants shows close structural similarities to the previously described mink cell focus-inducing (MCF)-type viruses.     

Persistence and recolonisation determine success of bog restoration for aquatic invertebrates: a comment on Mazerolle et al. (2006)

1. Mazerolle et al. (2006) concluded that some aquatic invertebrate species, including bog‐associated species, readily colonise man‐made bog pools. In contrast, in Dutch bog remnants Van Duinen et al. (2003) found that a considerable number of bog‐associated species do not colonise newly created bog pools. 2. The conclusion of Mazerolle et al. (2006) is based on vagile aquatic invertebrates. Here, we question whether their conclusion can be extended to more sedentary species, which were not captured in the Canadian study, but made up an important part of the invertebrate assemblage in the Dutch study. This discrepancy could be caused by sampling artefacts, low colonisation rates of these species or an incomplete restoration of site conditions. 3. In Canada, chances of recolonisation may be higher than in the Netherlands, as natural and near‐natural bogs are more extensive. In the Netherlands, with low chances of recolonisation, persistence of species may be more important. To disentangle the relative importance of persistence and recolonisation, evaluations of the success of restoration projects need to cover the entire invertebrate assemblage, including both vagile and more sedentary species.     

In vivo control of Mycosphaerella pinodes on pea leaves by a crude bulb extract of Eucomis autumnalis

We previously reported on the in vitro antifungal activity of a crude whole plant extract from Eucomis autumnalis against seven economically important plant pathogenic fungi. A crude extract of the bulb showed similar in vitro mycelial growth inhibition of the same plant pathogenic fungi as well as that of an eighth fungus, Mycosphaerella pinodes, the cause of black spot or Ascochyta blight, in peas. Subsequently, fourth internode leaves were removed from 4 wk old pea plants, placed on moist filter paper in Petri dishes and inoculated with an M. pinodes spore suspension before and after treatment with the extract. The control of Ascochyta blight by different concentrations of the crude E. autumnalis extract was followed in vivo by leaf symptoms over a 6 day period at 20°C in a growth cabinet. The crude extract prevented M. pinodes spore infection of the leaves when the leaves were inoculated with spores both before or after treatment with the extract, confirming complete inhibition of spore germination. The crude E. autumnalis extract showed no phytotoxic reaction on the leaves even at the highest concentration applied.     

Measurement of mass flow of water in the stems of herbaceous plants

Abstract. Heat balance methods of stem flow measurement offer the opportunity to measure directly the mass flow rate of water in plants. We have tested one such approach; the constant power heat balance method of Sakuratani (1981). The results supported his statement of an approximate accuracy of 10% when measuring the transpiration rate of herbaceous plants. The response to sudden changes in stem flow rate is not instantaneous, but investigation of the time constant shows that it decreases as stem flow increases, to the extent that, at flow rates typical of daytime conditions the system is capable of accurately tracking changes in stem flow within 5 min or less.
We describe a new gauge design that is relatively rugged, simple to use with an appropriate digital datalogger and suitable for field use over prolonged periods of time. It does not injure or penetrate the stem, is amenable to continuous and direct recording of the mass flow rate of water in the stem and requires no calibration. A further refinement, which should improve both the accuracy and the dynamic response of the system, is proposed.     

Antennal sensory structures in Nepticulidae (Lepidoptera) and their phylogenetic implications

Antennae of representatives of most genera and subgenera of Nepticulidae have been examined in detail with SEM. Flagellar segments are highly uniform and possess each 1–3 s. trichodea, 1–3 long s. chaetica type I, 12 short s. chaetica type II and 0–2 s. coeloconica of a simpified type without “picket fence”. The dominating structure is a pair of s. vesiculoclada, a unique sensillum type, believed to be a synapomorphy for the Nepticulidae. A pair is present on alt flagellomeres in male antennae but not on all flagellomeres of females in two subgenera of Ectoedemia. Principally each s. vesiculocladum is five-branched, with the branches joined tightly to the flagellar cuticle, but reduction of the branches occurs in several genera, in particular in Ectoedemia. This sensillum can be classified as a single-walled wp sensillum, and thus a chemoreceptor. The s. vesiculocladum is considered to be a homologue of the ascoid sensilla in the Opostegidae. The presence of these special setae is considered as an additional synapomorphy for both families. A review of previous Literature on lepidopteran sensilla is presented and the present data are compared and discussed. In the Nepticulidae the s. vesiculocladum is assumed to be the major pheromone detector in contrast to the s. trichodea of most other Lepidoptera.     

Seasonal changes in the pigments, carbohydrates and growth of red spruce as affected by ozone

Two-year-old red spruce seedlings were exposed to various levels ot ozone, from 0.4 to 3 times ambient levels, in open-top chambers in Ithaca, NY, USA. Exposures, which varied with changes in day length, commenced on May 30, 1987 and continued until December 14, 1987. Seedling biomass, carbohydrate contents, pigment contents, and rate of electron transport were assessed twice monthly during the fumigation period. Orthogonal quadratic or cubic polynomials were used to model the response through time each variable measured. A one-way analysis of variance model was fitted to every regression coefficient in each polynomial model to test for ozone effects on seasonal physiological patterns. Ozone did not influence growth, foliar pigment content, foliar starch content, root carbohydrate content, or rate of electron transport. The seasonal change of needle raffinose content differed between exposed to low (0.4 ×, 1×) and high (2×, 3×) ozone levels. There was also a trend towards reduced total soluble sugar content foliage during late autumn in higher ozone treatments.     

Structural comparison of the 68 kDa laminin-binding protein and 5'-nucleotidase from chicken muscular sources: evidence against a gross structural similarity of both proteins

The 68 kDa laminin-binding protein purified from chicken skeletal muscle and the ectoenzyme 5'-nucleotidase from chicken gizzard are both able to interact with laminin. They were both shown to possess a nearly identical amino acid composition. The 79 kDa glycosylated form of 5'-nucleotidase can be transformed into an enzymatically active form by treatment with endoglycosidase F (Endo F). Deglycosylated (Endo F-treated) 5'-nucleotidase exhibits an apparent molecular mass of 68 kDa. Using immunological and finger-printing techniques, both proteins were analysed to determine their structural relatedness. The results obtained indicate that both proteins are not identical but may posses a few common peptides of yet unknown sequence and length.