Effect of amphotericin B on the lipids of five different strains ofCryptococcus neoformans

Cells of five strains ofCryptococcus neoformans were obtained for partial analysis of lipid composition. Quantitative analysis of lipids and sterols were completed, as well as qualitative analysis of sterols by thin-layer chromatography and by the ultraviolet spectra. Such determinations were made on cells cultured in the absence and presence of amphotericin B at sub-MIC (minimal inhibitory concentration) levels. Marked alterations of the lipid and sterol contents were observed in the amphotericin B — treated cells. Moreover, ergosterol disappeared in these antibiotic-exposed cells. It is concluded that amphotericin B altered the lipid profiles, especially sterols ofC. neoformans.     

Carbohydrate and lipid components of hyphae and conidia of human pathogenFonsecaea pedrosoi

The carbohydrate and lipid components of mycelium and conidia ofFonsecaea pedrosoi (Brumpt) were analysed by paper, thin-layer and gas-chromatography, mass spectrometry and ultraviolet spectroscopy. Glucose, mannose, galactofuranose, rhamnose and glucosamine were polysaccharide components identified inF. pedrosoi. Significant changes in the carbohydrate pattern occurred during the conversion of mycelium into conidia. Rhamnose was predominant in conidia whereas galactose was prominent in mycelium. Palmitic, stearic, oleic, linoleic, and arachidonic acids were the fatty acids identified in the total lipid fraction. Palmitic and oleic acids were major fatty acids. Marked alterations in the fatty acid constituents were observed between the cell types ofF. pedrosoi. Arachidonic acid was detected only in conidia and linoleic acid was preferentially identified in mycelium. Differences in the sterol composition was also associated with morphogenesis inF. pedrosoi. Two main sterols, ergosterol and another less polar sterol, not fully characterized, were found in mycelium whereas in conidia only the latter sterol was present.     

Seasonal patterns of total and energy reserve lipids of dominant zooplanktonic crustaceans from a hyper-eutrophic lake

Summary Seasonal patterns of lipid reserves and lipid classes of dominant zooplankton in a hyper-eutrophic lake were examined in relation to algal food resources. Triacylglycerol was the principle lipid energy reserve in all five species examined. During the height of the yearlyAphanizomenon flos-aquae bloom, lipid levels of the principle herbivores (Daphnia pulex andLeptodiaptomus sicilis) and an omnivore (Diacyclops bicuspidatus thomasi), were at their lowest concentration, suggesting that this cyanobacterium is nutritionally inadequate. As the cyanobacterial bloom began to collapse, bacterial numbers increased rapidly. The increase in bacterial numbers coincided with a large increase in areal lipid energy reserves ofDiaphanosoma leuchtenbergianum andChydorus sphaericus. Examination of seasonal patterns in the biomass of different algal species suggested thatRhodomonas minuta andCryptomonas erosa played a key role in nutrition, lipid deposition, and reproduction ofD. pulex andL. sicilis.     

Chloroplast alterations induced by tomato bushy stunt virus inDatura leaves

Summary Peculiar chloroplast alterations were found in mesophyll cells ofDatura stramonium systemically infected with tomato bushy stunt virus. These alterations lead to complete rearrangement of the thylakoids.     

Beeinflussung der Blattzellen vonFontinalis antipyretica durch den Bewuchs vonCocconeis placentula

In a special habitat near Lunz (Mittersee) long time epiphytism ofCocconeis placentula var.tenuistriata and var.klinoraphis onFontinalis antipyretica leaves causes characteristic, strongly localized alterations in the walls of affected cells. They turn brown, may thicken, and may produce cystolithe-like protuberances into the cell lumina. These reactions obviously are brought about by a specific property ofCocconeis, for other epiphytes do not cause any change. The comparison of theCocconeis vegetation on younger and olderFontinalis leaves shows a considerable variability of the reactions mentioned, and in another habitat none could be found. Apparently several factors cause these reactions. Altogether,Cocconeis placentula is not a completely harmless epiphyte under certain circumstances. The alterations of the moss cells may be interpreted partly as a premature aging process, partly as a defensive action corresponding to alterations induced by some parasitic fungi. Furthermore, the reactions ofCocconeis onFontinalis are remarkably parallel to the influence of the dinococcalean algaRaciborskia inermis onSpirogyra grevilleana.

     

Characterization of transgenic plants derived from hairy roots ofHyoscyamus muticus

Mature plants were regenerated via protoplasts fromAgrobacterium rhizogenes-transformed root cultures ofHyoscyamus muticus L., and chemical analyses were performed on 34 individual plants. The regenerated plants showed strong phenotypic differences from clone to clone as well as from the control plants. Polymerase chain reaction studies revealed that the plants exhibiting the strongest phenotypic alterations contained therol (A, B and C) genes, whereas the plants with fewer alterations had lost them. The plants produced hyoscyamine, scopolamine and a range of different calystegins, and considerable somaclonal variation was observed. Alkaloid production in the plants transgenic for therol genes was clearly reduced. The pattern of calystegins was similar within all the regenerated plants lackingrol genes. Among the plants withrol genes, the calystegin B₁ was not detectable. It seems clear that the presence ofrol genes is detrimental to the alkaloid accumulation in the transgenic plants in contrast to hairy root cultures.Abbreviation PCR Polymerase chain reaction     

Effect of amphotericin B on the lipids of yeast cells of Sporothrix schenckii

Yeast cells of five strains of Sporothrix schenckii were obtained for partial analysis of lipid composition. Quantitative analysis of lipids and sterols were completed, as well as qualitative analysis of sterols by thin layer chromatography and by ultraviolet spectra. These determinations were made on cells cultured in the absence and presence of amphotericin B at sub-MIC (minimum inhibitory concentration) levels. Marked alterations in lipid content were observed in the amphotericin B-treated cells. The major alterations were the reduction of total lipid (18.7–57.6%) and sterols (48.5–96.7%) after exposure to the polyenic antibiotic. It is concluded that amphotericin B altered the lipid profiles, especially sterols of S. schenckii.     

Study on the innocuity ofHirsutella thompsonii. I. Infectivity in mice and guinea pigs

The present work concerns the innocuity ofHirsutella thompsonii Fisher isolated in Cuba from a citrus rust mite,Phyllocoptruta oleivora Ashm. This phytopathogenic arachnide is an important pest of different citrus fruit plantations. The infectivity tests were performed in 220 animals (mice and guinea pigs) inoculated by intraperitoneal and subcutaneous injections. Animals showed macroscopic alterations such as nodules in the liver capsule, spleen, subcutaneous tissue; superficial fibrinous adhesions on these organs and hepatosplenomegaly. No fungi were recovered from organs at any time and only disintegrated hyphal elements were observed. The histopathological study disclosed a non-specific tissue reaction to a foreign body. The results obtained suggest the non-infectivity and innocuity ofHirsutella thompsonii Fisher in mice and guinea pigs.      

Distribution of alkanes and fatty acids in theCondalia montana (Rhamnaceae) species complex

The alkane and fatty acid composition of the lipid extracts from leaves of 14 populations ofCondalia: C. microphylla f.xanthocarpa, C. microphylla f.erythrocarpa, C. microphylla f.melanocarpa, C. montana, andC. buxifolia, were determined by gas chromatography. Seventeen alkanes and 14 fatty acids were investigated and a predominance of nonacosane, hentriacontane, palmitic and linoleic acids in all species was found.     

Co-suppression in transgenicPetunia hybrida expressing chalcone synthase A (chsA)

Chalcone synthase A is a key enzyme in the anthocyanin biosynthesis pathway. Expression ofchsA gene in transgenicPetunia hybrida resulted in flower color alterations and co-suppression of transgenes and endogenous genes. We fused the β-glucuronidase (uidA) gene to the C-terminal ofchsA gene, and transferred the fusion gene intoPetunia hybrida viaAgrobacterium tumefaciens. GUS histochemical staining analysis showed that co-suppression occurred specifically during the development of flowers and co-suppression required the mutual interaction of endogenous genes and transgenes. RNAin situ hybridization analysis suggested that co-suppression occurred in the entire plant, and RNA degradation occurred in the cytoplasm.     

Chromosome alterations in tissue culture cells ofAllium fistulosum

Heterochromatin behaviour and structural alterations in chromosomes of cells derived from callus culture ofAllium fistulosum have been studied.The diploid chromosome complement ofAllium fistulosum consists of 16 chromosomes with significant amount of heterochromatin mainly of telomeric nature. In eight collections of callus cells analysed, a high rate of numerical and structural chromosome abnormalities was observed. After 12 months in culture about 20% of metaphase chromosomes possessed distinct signs of mutational events.C-banded preparations revealed that many structural alterations involved regions of heterochromatin. Interchromosomal connections and chromatid fusions occurred at telomeric heterochromatin segments. Also formation of the end-to-end associations and polycentric chromosomes often took place without visible loss of telomeric heterochromatin.     

Cytotaxonomical studies of West Bengal charophyta: Karyotype analysis in Chara braunii

The paper deals with cytotaxonomic studies of three forms ofChara braunii-f.coromandelina (n = 14), f.schweinitzii (n = 14) and f.kurzii (n = 28) from West Bengal (India). An improved technique involving the use of a pretreating agent permitted a detailed analysis of the karyotypes of the three taxa studied. It has been concluded that structural alterations of chromosomes and polyploidy are associated with evolution of phenotypic differences in theChara braunii complex. The phylogenetic status ofChara braunii in the light of the present investigation is discussed.     

Biophysical characterization of OprB,a glucose-inducible porin ofPseudomonas aeruginosa

OprB, a glucose-inducible porin ofP. aeruginosa, was characterized by black lipid bilayer analysis and circular dichroism spectroscopy. Black lipid bilayer analysis of OprB revealed a single-channel conductance of 25 pS, the presence of a glucose binding site with aK _s for glucose of 380 ± 40 mM, and the formation of channels with a strong selection for anions. Analysis ofP. aeruginosa OprB circular dichroism spectra revealed a high sheet content (40%) which is within the range of that determined for other porins. Values obtained from black lipid bilayer analysis were compared to those previously obtained for OprB ofP. putida [Saravolacet al. (1991).J. Bacteriol. 173, 4970–4976] and indicated extensive similarities in the single-channel conductance and glucose-binding properties of these two porins. Immunological and amino terminal sequence analysis revealed a high degree of homology. Of the first 14 amino terminal residues, 12 were identical. A major difference between the two porins was found in their ion selectivity. WhereasP. aeruginosa OprB is anion selective,P. putida OprB and other carbohydrate selective porins are known to be cation selective.     

Phospholipids of ethambutol-susceptible and resistant strains ofMycobacterium smegmatis

The composition, subcellular distribution and rate of synthesis of phospholipids were compared in ethambutol susceptible and resistant strains ofMycobacterium smegmatis. Significant quantitative alterations in phospholipids accompanied the acquisition of resistance, whereas fatty acyl group composition of total phospholipid remained the same in ethambutol resistant and susceptible strains. Cell wall of resistant strain exhibited an accumulation of phospholipids and a decrease in the degree of unsaturation of phospholipid fatty acyl groups. Changes in the cell wall phospholipid composition may contribute to resistance ofMycobacterium smegmatis to ethambutol.     

Uninfected red cells from malaria-infected blood: Alteration of fatty acid composition involving a serum protein: An in vivo and in vitro study

Summary Alteration of uninfected erythrocytes, fromPlasmodium (the malaria parasite)-infected blood remained an open question. In this study we compared the in vivo fatty acid compositions of control and uninfected monkey erythrocytes. A large (40%) increase in the linoleic acid level was observed, which was recovered mostly in neutral lipids. An in vitro system was developed to study medium-mediated alterations of cultured erythrocytes byPlasmodium falciparum. The increase in the linoleate level was reproduced in vitro and was also localized in the neutral lipid fraction, especially in triacylglycerols. Studies using proteolytic digestion and heat denaturation showed that a heat-labile serum protein is indispensable for the increase in the linoleate level of red cells treated with the supernatant, ofP. falciparum cultures. Both the function and the mechanism of this modification of uninfected erythrocytes still remain unknown. This work was supported by the UNDP/World Bank/WHO special program for Research and Training in Tropical Diseases (grant T16-181-M2-15B).     

Phylogenetic relationships of JapaneseAster (Asteraceae, Astereae) sensu lato based on chloroplast-DNA restriction site mutations

RFLPs of cpDNA were examined for 18 species ofAster, six species ofKalimeris, two species ofMiyamayomena and one species and one variety ofHeteropappus from Japan, using 16 restriction endonucleases. Approximately 275 restriction sites were surveyed, and a total of 74 restriction site mutations was detected, and 31 of these were phylogenetically informative. Sixteen most parsimonious trees constructed from Wagner parsimony analysis indicated the polyphyly ofKalimeris andMiyamayomena sensu Kitamura;K. miqueliana belongs to a different clade from the remaining species ofKalimeris, and two species ofMiyamayomena did not make a single clade. This result suggests that the shortening or loss of pappus have happened parallelly in different evolutionary lineages. We must be careful to assess the pappus character in taxonomy and phylogeny, and it is desirable to examine their phylogenetic relationships using a molecular data.     

A comparative biochemical study of conifer pollen allergens

In the order Coniferales, only the family Cupressaceae is regarded as being a significant source of airborne allergens, withJuniperus ashei characterized as the most significat aeroallergen. Pollen of the closely related speciesJ. virginiana has been shown to cross-react withJ. ashei pollen, however,J. virginiana pollen is not considered an important aeroallergen. Although there have been several reports of allergies toPinus pollen, the pollen of this genus is regarded as hypoallergenic. Our previous studies have shown that pollen extracts ofJ. ashei, J. virginiana, J. pinchotii, Cupressus macrocarpa, Pinus echinata andP. taeda all contained several proteins with the same molecular weights including the reported allergen ofJ. ashei. The present study compared the biochemistry ofJ. ashei, J. virginiana andP. echinata pollen. A time course experiment ofJ. ashei, J. virginiana andP. echinata showed thatJ. ashei released a greater quantity of protein within the first minute of moistening. SDS-PAGE analyses showed that the reported allergen ofJ. ashei pollen extracts was released in large quantities within the first minute of extraction. It was also determined that individual pollen grains ofP. echinata contained a greater quantity of protein than the pollen ofJ. ashei andJ. virginiana, but due to the large size of pine pollen there was less protein per gram of pollen. Lipid analysis of these three taxa showed that the pollen ofP. echinata contained more lipid per grain and per gram of pollen. Results indicate that the rapid release of the reported allergen fromJ. ashei pollen contributes to the allergenicity of this species compared to bothJ. virginiana andP. echinata.     

Variation in adult life history and stress resistance across five species of<Emphasis Type="Italic">Drosophila</Emphasis>

Dry weight at eclosion, adult lifespan, lifetime fecundity, lipid and carbohydrate content at eclosion, and starvation and desiccation resistance at eclosion were assayed on a long-term laboratory population ofDrosophila melanogaster, and one recently wild-caught population each of four other species ofDrosophila, two from themelanogaster and two from theimmigrans species group. The relationships among trait means across the five species did not conform to expectations based on correlations among these traits inferred from selection studies onD. melanogaster. In particular, the expected positive relationships between fecundity and size/lipid content, lipid content and starvation resistance, carbohydrate (glycogen) content and desiccation resistance, and the expected negative relationship between lifespan and fecundity were not observed. Most traits were strongly positively correlated between sexes across species, except for fractional lipid content and starvation resistance per microgram lipid. For most traits, there was evidence for significant sexual dimorphism but the degree of dimorphism did not vary across species except in the case of adult lifespan, starvation resistance per microgram lipid, and desiccation resistance per microgram carbohydrate. Overall,D. nasuta nasuta andD. sulfurigaster neonasuta (immigrans group) were heavier at eclosion than themelanogaster group species, and tended to have somewhat higher absolute lipid content and starvation resistance. Yet, these twoimmigrans group species were shorter-lived and had lower average daily fecundity than themelanogaster group species. The smallest species,D. malerkotliana (melanogaster group), had relatively high daily fecundity, intermediate lifespan and high fractional lipid content, especially in females.D. ananassae (melanogaster group) had the highest absolute and fractional carbohydrate content, but its desiccation resistance per microgram carbohydrate was the lowest among the five species. In terms of overall performance, the laboratory population ofD. melanogaster was clearly superior, under laboratory conditions, to the other four species if adult lifespan, lifetime fecundity, average daily fecundity, and absolute starvation and desiccation resistance are considered. This finding is contrary to several recent reports of substantially higher adult lifespan and stress resistance in recently wild-caught flies, relative to flies maintained for a long time in discretegeneration laboratory cultures. Possible explanations for these apparent anomalies are discussed in the context of the differing selection pressures likely to be experienced byDrosophila populations in laboratory versus wild environments. This paper is dedicated to the memory of our friend and former colleague Dr Hans Raj Negi, who tragically passed away at a very young age in a road accident in November 2003.     

A phylogenetic analysis ofDuguetia (Annonaceae) based on morphological data

In this paper we present a phylogeny based on a study of the general morphology of 61 of the ca. 80 species ofDuguetia. Analyses were performed with PAUP. The resulting phylogeny is compared with the classification ofDuguetia as suggested byFries (1934, 1937, 1939, 1959). The results are partly corresponding withFries's classification. SectionAlcmene includingD. rionegrensis, and sects.Geanthemum, Synsepalantha, andCalothrix withoutD. uniflora, are upheld by this analysis. It is concluded that sect.Xylopipetalum should be united with sect.Duguetia. Before other taxonomic decisions at the section level can be made, more solid data must be available. Most dissimilarities found are considered to be not strong enough to propose alterations.Studies inAnnonaceae XXIII. For 22nd part seeZuilen & Maas (1994).