Aluminum toxicity studies in Vaucheria longicaulis var. macounii (Xanthophyta, Tribophyceae). I. Effects on cytoplasmic organization

Using differential interference contrast (DIC) and epifluorescence microscopy, we tested the hypothesis that exposure to environmentally significant levels of aluminum (Al) would cause rapid changes in cytoplasmic organization in vegetative filaments of the coenocytic alga, Vaucheria longicaulis Hoppaugh var. macounii Blum resulting in the loss of cytoplasmic streaming. In untreated cells, DIC microscopy revealed the presence of cortical cytoplasmic strands that were oriented longitudinally to the cell axis as well as sub-cortical cytoplasmic strands that exhibited a reticulate morphology. Organelles such as chloroplasts and mitochondria translocated throughout the cell in close association with the cortical longitudinal cytoplasmic strands. Staining with the lipophilic dye, 3,3-dihexyloloxacarbocyanine, revealed structures that appeared to be endoplasmic reticulum (ER). This organelle closely resembled, in location and appearance, the cytoplasmic strands visualized using DIC microscopy. The addition of Al (80 μM) resulted in the inhibition of cytoplasmic streaming as well as the dissipation of the putative cortical longitudinal ER within one minute. Subsequently, the DIC-visible cortical cytoplasmic strands exhibited progressive degrees of disorganization. Throughout these changes, chloroplasts and mitochondria remained visibly associated with the cortical cytoplasmic strands.     

Organization of the cytoskeleton in the coenocytic algaVaucheria longicaulis var.macounii: an experimental study

Summary The organization of the cytoskeleton of vegetative filaments ofVaucheria longicaulis Hoppaugh var.macounii Blum is investigated by fluorescence microscopy using monoclonal anti -tubulin antibodies and fluorescein (FITC)-labelled phalloidin. Confocal laser scanning microscopy observations give further information on the distribution of the cytoskeletal elements. Phalloidin labelling reveals F-actin bundles in the cortical cytoplasm of both fixed and unfixed vegetative filaments of this alga. In addition a more diffuse fluorescent component, seen at higher magnification to be made up of thinner F-actin bundles, can also be detected in unfixed cells. The distribution of the F-actin bundles resemble that of filamentous structures observed with differential interference contrast (DIC) microscopy in living cells. These structures seem to correspond to the microtubule associated reticulum (MAR) described in literature and overall the evidence suggests that actin and MAR elements are co-distributed. F-actin bundles are always found in association with focal masses (foci) of phalloidin-positive material. Foci are also observed by DIC microscopy associated with the cytoplasmic filamentous structures in living cells.Depolymerization of F-actin with cytochalasin D and the subsequent repolymerization that occurs on transfer ofVaucheria vegetative filaments to cytochalasin-free medium suggest that these foci are involved in the organization of the F-actin array. Immunofluorescence for -tubulin reveals microtubule bundles that are shorter in length and straighter in configuration than microfilament bundles. Microtubule bundles are associated with spot-like focal structures that, in many instances, show a close relationship with respect to nuclei. Oryzalin and cold temperature cause the depolymerization of the microtubule bundles and suggest, in conjunction with repolymerization studies, that these fluorescent spots associated with the ends of the microtubule bundles are involved in their organization; hence, they represent microtubule organizing centres or MTOCs. The importance of both microfilament and microtubule bundle focal regions is discussed with respect to the apical growth exhibited by the vegetative filaments of this alga.     

Effects of chemical speciation in growth media on the toxicity of mercury(II).

The toxicity of metals, including mercury, is expressed differently in different media, and the addition of soluble organics to the growth medium can have a significant impact on bioassay results. Although the effect of medium composition on metal toxicity is generally attributed to its effect on metal speciation (i.e., the chemical form in which the metal occurs), the importance of individual metal-ligand species remains largely unclear. Here, we report the results of a study that investigated, both experimentally and from a modeling perspective, the effects of complex soluble organic supplements on the acute toxicity (i.e., 50% inhibitory concentration [IC50]) of mercury to a Pseudomonas fluorescens isolate in chemically well-defined synthetic growth media (M-IIX). The media consisted of a basal inorganic salts medium supplemented with glycerol (0.1%, vol/vol) and a variety of common protein hydrolysates (0.1%, vol/vol), i.e., Difco beef extract (X = B), Casamino Acids (X = C), peptone (X = P), soytone (X = S), tryptone (X = T), and yeast extract (X = Y). These were analyzed to obtain cation, anion, and amino acid profiles and the results were used to compute the aqueous speciation of Hg(II) in the media. Respirometric bioassays were performed and IC50s were calculated. Medium components varied significantly in their effects on the acute toxicity of Hg(II) to the P. fluorescens isolate. IC50s ranged from 1.48 to 14.54 micrograms of Hg ml-1, and the acute toxicity of Hg(II) in the different media decreased in the order M-IIC >> M-IIP > M-IIB >> M-IIT > M-IIS >>> M-IIY.(ABSTRACT TRUNCATED AT 250 WORDS)     

Observations on the Structure of the Zoospores of Vaucheria, II

Sections of Vaucheria zoospores fixed at various stages havebeen examined in the electron microscope. Changes in the organizationof the cytoplasm and the redistribution of its organelles aredescribed and correlated with observations of living sporesduring their transformation into vegetative cells. In particular,proof is given that the disappearance of the cilia involvesthe withdrawal of the fibrillar axes of these organs into thecytoplasm and that the disappearance of the transparent outerzone of the motile spore involves the inward movement of specialcytoplasmic vesicles which form a main component of this layer. As a result of improved fixation new structural details aregiven, the most important of which concern the cytoplasmic matrix,the boundary membranes of the nuclei, the golgi bodies and theirrelation to mitochondria, and the internal structure of thechloroplasts. With the aid of serial sections it is shown thatthe internal laminae of the chloroplast are connected with eachother in a special manner.     

Ethylene thiourea (ETU). A review of the genetic toxicity studies

Ethylene thiourea (ETU) is a common contaminant, metabolite and degradation product of the fungicide class of ethylene bisdithiocarbamates (EBDCs); as such, they present possible exposure and toxicological concerns to exposed individuals. ETU has been assayed in many different tests to assess genotoxicity activity. While a great number of negative results are found in the data base, there is evidence that demonstrates ETU is capable of inducing genotoxic endpoints. These include responses for gene mutations (e.g. Salmonella), structural chromosomal alterations (e.g. aberrations in cultured mammalian cells as well as a dominant lethal assay) and other genotoxic effects (e.g. bacterial rec assay and several yeast assays).It is important to consider the magnitude of the positive responses as well as the concentrations/doses used when assessing the genotoxicity of ETU. While ETU induces a variety of genotoxic endpoints, it does not appear to be a potent genotoxic agent. For example, it is a weak bacterial mutagen in the Salmonella assay without activation in strain TA1535 at concentrations generally above 1000 μg/plate. Weak genotoxic activity of this sort is usually observed in most of the assays with positive results. Since ETU does not appear very potent and is not extremely toxic to test cells and organisms, it is not surprising to find that ETU does not produce consistent effects in many of the assays reviewed. Consequently, in many instances, mixed results for the same assay type are reported by different investigators, but as reviewed herein, these results may be dependent upon the test conditions in each individual laboratory. A primary shortcoming with many of the reported negative results is that the concentrations or doses used are not high enough for an adequate test for ETU activity. There are also problems with many of the negative assays generally in protocol or reporting, particularly with the in vivo studies (e.g. inappropriate sample number and/or sampling times; inadequate top dose employed).Overall, while ETU does not appear to be a potent genotoxic agent, it is capable of producing genotoxic effects (e.g. gene mutations, structural chromosomal aberrations). This provides a basis for weak genotoxic activity by ETU. Furthermore, based on a suggestive dominant lethal positive result, there may be a concern for heritable effects. Due to the many problems with the conduct and assessment of the in vivo assays, it is worth repeating in vivo     

Physicochemical studies on xylinan (acetan). II. Characterization by static light scattering

Laboratory-made samples of the polysaccharide xylinan, also called acetan, were studied in aqueous solution at various ionic strengths I (0.01 mol/L ≤ I ≤ 0.30 mol/L). The conditions for clarification (ultracentrifugation/membrane filtration) were studied. The Zimm procedure was used to obtain the average molar mass, the second virial coefficient, and the radius of gyration. The interpretation of the angular dependence of scattered light by fitting with “Master Curves” led to double-stranded wormlike chains with persistence lengths between 90 and 120 nm. The ionic strength had a strong effect on the thermodynamic second virial coefficient, but the overall structure remained unchanged. The ambiguity of the light scattering data was discussed assuming alternatively a two-component system instead of the wormlike chain model for the experimental scattering curves. The two-component system can be ruled out on the basis of model calculations. © 1996 John Wiley & Sons, Inc.     

Effects of manganese toxicity on photosynthesis of white birch (Betula platyphylla var. japonica) seedlings

The effects of manganese (Mn) toxicity on photosynthesis in white birch ( Betula platyphylla var. japonica ) leaves were examined by the measurement of gas exchange and chlorophyll fluorescence in hydroponically cultured plants. The net photosynthetic rate at saturating light and ambient CO₂ (C_a) of 35 Pa decreased with increasing leaf Mn concentrations. The carboxylation efficiency, derived from the difference in CO₂ assimilation rate at intercellular CO₂ pressures attained at C_a of 13 Pa and O Pa, decreased with greater leaf Mn accumulation. Net photosynthetic rate at saturating light and saturating CO₂ (5%) also declined with leaf Mn accumulation while the maximum quantum yield of O₂ evolution at saturating CO₂ was not affected. The maximum efficiency of PSII photochemistry (F_v/F_m) was little affected by Mn accumulation in white birch leaves over a wide range of leaf Mn concentrations (2–17 mg g₋₁ dry weight). When measured in the steady state of photosynthesis under ambient air at 430 μmol quanta m₋₂ s₋₁, the levels of photochemical quenching (qP) and the excitation capture efficiency of open PSII (F'^v/F'^m) declined with Mn accumulation in leaves. The present results suggest that excess Mn in leaves affects the activities of the CO² reduction cycle rather than the potential efficiency of photochemistry, leading to increases in Q_A reduction state and thermal energy dissipation, and a decrease in quantum yield of PSII in the steady state.     

Biosystematic studies on the umbellatum-angustifolium complex of the genus Ornithogalum. (Liliaceae). II. Genome characterization and evolution

Chromosome banding techniques (Giemsa and fluorochrome staining) were used to investigate Ornithogalum umbellatum L., O. angustifolium Bor. and 11 related taxa. Nine different basic Q-banding patterns, referred to as genomes, could be recognised. These patterns are indicated by capitals (A–E, P–S). Six of the nine basic genome types can be subdivided into different subtypes. The B genome was found in the two bulbilliferous species, i.e. O. umbellatum and O. angustifolium. It occurs in a number of combinations together with the A, C, D and E genomes. Genome combinations of these five basic types were found in seven species. Six of them share morphological characteristics. These species are: O. monticolum Jord. & Fourr., O. baeticum Boiss., O. angustifolium, O. refractum, Kit., O. umbellatum and O. algeriense Jord. & Fourr. A seventh species, O. platyphyllum Boiss., in which the E genome occurs, can be regarded as an intermediate between this complex and the O. montanum group. Each species is treated extensively; its morphology, geographical distribution, chromosome number and banding patterns are discussed separately. A model of the probable course of the evolution in this complex during the last two glaciation periods is described. In this model attention is paid to ecological differentiation, migration leading to the present distribution patterns, hybridization followed by polyploidization and to fixation, which may be the result of chromosomal rearrangements that prevent regular meiosis. Some cytotypes, formerly regarded as O. umbellatum, are now classified in other species. A second polyploid complex in which these species occur, is probably more widely distributed in south-eastern Europe than is generally thought at present. The species O. exscapum Ten., O. nivale Boiss., O. comosum L., O. collinum Guss. and O. gussonei Ten. p.p. are components of this complex. The delimitation of this complex as a whole and the relationship between its diploid and polyploid components have still to be clarified.     

Further studies of the glucomannan from Aloe vahombe (liliaceae). II. Partial hydrolyses and NMR 13C studies

The polysaccharide from Aloe vahombe (lilaceae) was submitted to partial hydrolyses by sulfuric, oxalic and phosphoric acid. Some oligosaccharides were isolated and investigated by chemical and 13C NMR spectroscopic methods. Their structure was determined. The results prove unambiguously that in the analyzed material the D-mannose is linked to the D-glucose by beta-1----4 linkages to form a carbohydrate heteropolymer.     

Histochemical studies on Raillietina (Raillietina) johri (Cestoda: Davaineidae). II. Nucleoside diphosphatase and thiamine pyrophosphatase.

Thiamine pyrophosphatase and nucleoside diphosphatase have been studied histochemically in Raillietina (Raillietina) johri. Thiamine pyrophosphatase activity has been observed in the tegument, subtegumental muscle, subtegumental cells, medullary parenchyma, excretory canal and various reproductive structures like testes, ovary, vas deferens, spermatozoa and vitellaria. Eggs exhibit moderate enzyme activity. Various nucleoside diphosphates have been found to be hydrolyzed by thiamine pyrophosphatase. CaCl2, MgCl2 and MnCl2 each activated the enzyme at a final concentration of 6 mM whereas cysteine, reduced glutathione and PCMB inhibited the enzyme activity at a final concentration of 10 mM, 10 mM and 20 mM, respectively. KCN and NaF had no effect on the enzyme staining at concentration as high as 50 mM and 30 mM, respectively. Possible roles of the enzyme in the parasite have been discussed.