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.     

Poly(L-lysyl-L-alanyl-alpha-L-glutamic acid). II. Conformational studies.

The solution characterization of poly(Lys-Ala-Glu) is described. This polytripeptide is zwitterionic at neutral pH and is shown to take on a conformation which is dictated by the state of ionization, molecular weight, temperature, and solvent. The polypeptide is almost entirely α-helical at low pH and temperature for polymers of greater than 25,000 molecular weight. Melting profiles for these conditions show t_m ～ 20°C. Analysis of circular dichroism curves shows the α-helical content to vary in a linear manner with molecular weight in the range 3000–30,000. At neutral pH the charged polypeptide is essentially random, but substantial α-helix could be induced by addition of methanol or trifluoroethanol. At temperatures where the sequential polypeptide is a random coil, addition of trifluoroethanol produces a polymer which is mostly α-helical but also contains an appreciable ammount of β-structure. The infrared spectrum of a low-molecular-weight fraction assumed to be cyclo(Lys-Ala-Glu)₂ was tentatively assigned a β-pleated sheet structure. A comparison of this polytripeptide in various ionization states with other polytripeptides containing L -alanine and L -glutamate or L -lysine shows the α-helix directing properties for the (uncharged) residues to lie in the order Ala > Glu > Lys.     

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.     

Studies on the Genus Vaucheria (Xanthophyceae) in Iraq

Eight species of Vaucheria from southern Iraq, V. dichotoma, V. sessilis, V. erythrospora, V. verticillata, V. hamata, V. woroniniana, V. synandra, and V. longicaulis were studied and described. All but V. sessilis and V. longicaulis are recorded for the first time from Iraq.     

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.     

Macrozooplankton studies in Kuwait Bay (Arabian Gulf). II. Distribution and composition of planktonic penaeidea

Six penaeideans were identified during a 12-month investigationof macrozooplankton in Kuwait Bay. The numerically dominantspecies included two sergestids, Lucifer hanseni and Acetesjaponicus. Of the two, L.hanseni appears to spawn in the bayduring summer months while, A.japonicus may concentrate itsspawning in coastal waters during late spring through fall.The most abundant penaeids were Parapenaeopsis stylifera andMetapenaeus spp., although Kuwait Bay may not be a major spawningarea for these species. However, the Khor al Sabiya, a river-likechannel northeast of Kuwait Bay, may serve as a nursery forat least two of the penaeids (Metapenaeus spp. and Penaeus semisulcatus)as well as for A.japonicus. Both P.stylifera and Metapenaeusspp. larvae were most abundant during late spring. Penaeus semisulcatuslarvae exhibited fall and spring-summer maxima but were neverlocally abundant.