Distribution of <Emphasis Type="Italic">Doronicum clusii</Emphasis> and <Emphasis Type="Italic">D. stiriacum</Emphasis> (Asteraceae) in the Alps and Carpathians

In the course of a taxonomic revision of the Doronicum clusii agg. (Asteraceae), we present an updated and commented distribution map of D. clusii (All.) Tausch and D. stiriacum (Vill.) Dalla Torre from the Alps and Carpathians, completed with information on biogeography and taxonomy. We show that D. stiriacum was erroneously indicated for the Carpathians south and southeast of the Rodna Mountains in Romania and for the Western Alps. In the Western Carpathians, it has only been rarely mentioned for the Nízke Tatry (Lower Tatras) and the Belianske Tatry (Belá Tatras) in Slovakia.     

Microwave assisted synthesis of (<Emphasis Type="Italic">E</Emphasis>)-1-(2-((1-benzyl-1<Emphasis Type="Italic">H</Emphasis>-1,2,3-triazol-4-yl)methoxy)phenyl)-3-(9-ethyl-9<Emphasis Type="Italic">H</Emphasis>-carbazol-3-yl)prop-2-en-1-ones and their antimicrobial activity

A novel series of 1,2,3-triazolo-carbazole chalcones were synthesized by the Claisen-Schmidt condensation reaction followed by the copper catalyzed Huisgen (2 + 3) cycloaddition reaction by conventional as well as microwave irradiation methods. The structures of the newly synthesized compounds were established on the basis of IR, ¹H NMR, ¹³C NMR, and mass spectral data. All the compounds were screened in vitro for antibacterial and antifungal activities.     

Synthesis of orthogonally protected (<Emphasis Type="Italic">3R</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)- and (<Emphasis Type="Italic">3S</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)-4,5-diamino-3-hydroxypentanoic acids

Summary.  The paper describes two methods of the synthesis of ethyl (3R,4S)- and (3S,4S)-4-[(benzyloxycarbonyl)amino]-5-[(tert-butyloxycarbonyl)amino]-3-hydroxypentanoates, useful for the syntheses of edeine analogs. Differently N-protected (S)-2,3-diaminopropanoic acid was used as a substrate in both procedures. The absolute configuration of newly generated asymmetric carbon atoms C-3 in β-hydroxy-γ,δ-diamino products was assigned by means of ¹H NMR spectroscopy after their transformation into corresponding piperidin-2-ones. Received May 24, 2002 Accepted October 10, 2002 Published online December 18, 2002 Acknowledgment The authors are indebted to the Faculty of Chemistry, Technical University of Gdańsk for financial support. Authors' address: Zbigniew Czajgucki, M. Sc., Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Technical University of Gdańsk, 11/12 Narutowicza St., 80-952 Gdańsk, Poland, Fax +48 58 347 11 44, E-mail: zmczaj@wp.pl     

Molecular evolution of <Emphasis Type="Italic">Phox</Emphasis>-related regulatory subunits for NADPH oxidase enzymes

Background  

The reactive oxygen-generating N ADPH ox idases (Noxes) function in a variety of biological roles, and can be broadly classified into those that are regulated by subunit interactions and those that are regulated by calcium. The prototypical subunit-regulated Nox, Nox2, is the membrane-associated catalytic subunit of the phagocyte NADPH-oxidase. Nox2 forms a heterodimer with the integral membrane protein, p22 phox, and this heterodimer binds to the regulatory subunits p47 phox, p67 phox, p40 phox and the small GTPase Rac, triggering superoxide generation. Nox-organizer protein 1 (NOXO1) and Nox-activator 1 (NOXA1), respective homologs of p47 phox and p67 phox, together with p22 phox and Rac, activate Nox1, a non-phagocytic homolog of Nox2. NOXO1 and p22 phox also regulate Nox3, whereas Nox4 requires only p22 phox. In this study, we have assembled and analyzed amino acid sequences of Nox regulatory subunit orthologs from vertebrates, a urochordate, an echinoderm, a mollusc, a cnidarian, a choanoflagellate, fungi and a slime mold amoeba to investigate the evolutionary history of these subunits.     

Xanthones and flavonoids from <Emphasis Type="Italic">Gentiana algida</Emphasis> Pall

This article is devoted to phytochemical investigation of xanthones and flavonoids from the Gentiana algida Pall. (cold gentian, the Gentianaceae family). The following xanthones and flavones have been isolated from the top of the gentian: bellidifolin, isobellidifolin, swerchirin, 1,5,8-tirhydroxy-3,4-dimethoxyxanthone, swertisin, and swertianolin (flavone-C-glycoside). The isolated compounds have been identified on the basis of their chemical conversions and by their UV, mass, ¹H, and ¹³C NMR spectra. Derivatives of 1,3,5,8-tetrahydroxyxanthone were mainly found in the top of the cold gentian.     

Duplicate Gene Evolution Toward Multiple Fates at the <Emphasis Type="Italic">Drosophila melanogaster HIP</Emphasis>/<Emphasis Type="Italic">HIP-Replacement</Emphasis> Locus

Hsc/Hsp70-interacting protein (HIP) is a rapidly evolving Hsp70 cofactor. Analyses of multiple Drosophila species indicate that the HIP gene is duplicated only in D. melanogaster. The HIP region, in fact, contains seven distinctly evolving duplicated genes. The regional duplication occurred in two steps, fixed rapidly, and illustrates multiple modes of duplicate gene evolution. HIP and its duplicate HIP-R are adaptively evolving in a manner unique to the region: they exhibit elevated divergence from other drosophilids and low polymorphism within D. melanogaster. HIP and HIP-R are virtually identical, share polymorphisms, and are subject to gene conversion. In contrast, two other duplicate genes in the region, CG33221 and GP-CG32779, are pseudogenes, and the chimeric gene Crg1 is subject to balancing selection. HIP and HIP-R are evolving rapidly and adaptively; however, positive selection is not sufficient to explain the molecular evolution of the region as a whole. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Presence of <Emphasis Type="Italic">C. albidus</Emphasis>, <Emphasis Type="Italic">C. laurentii</Emphasis> and <Emphasis Type="Italic">C. uniguttulatus</Emphasis> in Crop and Droppings of Pigeon Lofts (<Emphasis Type="Italic">Columba livia</Emphasis>)

Columba livia is an important reservoir and carrier of Cryptococcus neoformans, Cryptococcus uniguttulatus, Cryptococcus laurentii and Cryptococcus albidus. Upper digestive tract of this species is also known as a habitat for Cryptococcus neoformans. Given the increasing clinical interest of this microorganism, 331 swabs from crop and 174 dropping samples from pigeon lofts in Grand Canary Island have been studied. The obtained results show an extensive presence samples 81 positive (24.47%) of Cryptococcus spp. in analysed crops: 32 (9.66%) for C. neoformans, 24 (7.2%) for C. uniguttulatus, 23 (6.9%) for C. albidus and 2 (0.6%) for C. laurentii. In the same way, Cryptococcus spp was also isolated in 82 (47.13%), dropping samples: C. neoformans in 59 (33.9%), C. uniguttulatus, in 9 (5.17%), C. laurentii in 8 (4.59%) and C. albidus in 6 (3.44%) of the investigated samples, respectively. The cryptococcosis produced by species of cryptococci other than C. neoformans has become more important during the last decade, supporting the study on the role of pigeon in the epidemiology of this disease.     

Habitat,density, and spatial distribution of <Emphasis Type="Italic">Rivulus giarettai</Emphasis> (<Emphasis Type="Italic">Actinopterygii</Emphasis>, <Emphasis Type="Italic">Cyprinodontiformes</Emphasis>) in southeastern Brazil

We present data on the habitat, density, and spatial distribution of Rivulus giarettai, and discuss some biotic and abiotic variables related to its abundance in Free Flowing Waters (FFW) and Dam Reservoirs (DR) in palm grove (Mauritia flexuosa) marshes (Veredas) in Uberlandia, Minas Gerais, southeastern Brazil. The mean density (individuals/plot) of R. giarettai was about 13 times higher in FFW than in DR. In FFW, the density of R. giarettai was highest at intermediate amounts of substrate (plant mass) and it was positively rank-correlated with the depth, and the number of arthropods. Individuals occurred in an aggregated distribution. The aggregated pattern could be related to a concentration of individuals in microhabitats neither too much exposed nor completely saturated by plants. R. giarettai was relatively abundant and tolerant to slight man-made habitat modifications. Damming appeared to be especially problematic by negatively affecting its density.     

Retrotransposon <Emphasis Type="Italic">gtwin</Emphasis>: Structural analysis and distribution in <Emphasis Type="Italic">Drosophila</Emphasis> strains

A search for noncanonical variants of the gypsy retrotransposon ( MDG4 ) in the genome of the Drosophila melanogaster strain G32 led to the cloning of four copies of the poorly studied 7411-bp gtwin element. Sequence analysis showed that gtwin belongs to a family of endogeneous retroviruses, which are widespread in the Drosophila genome and have recently been termed insect erantiviruses. The gtwin retrotransposon is evolutionarily closest to MDG4, as evident from a good alignment of their nucleotide sequences including ORF2 (the pol gene) and ORF3 (the env gene), as well as the amino acid sequences of their protein products. These regions showed more than 75% homology. The distribution of gtwin was studied in several strains of the genus Drosophila. While strain G32 contained more than 20 copies of the element, ten other D. melanogaster strains carried gtwin in two to six copies per genome. The gtwin element was not detected in D. Hydei or D. Virilis. Comparison of the cloned gtwin sequences with the gtwin sequence available from the D. melanogaster genome database showed that the two variants of the mobile element differ by the presence or absence of a stop codon in the central region of ORF3. Its absence from the gtwin copies cloned from the strain G32 may indicate an association between the functional state of ORF3 and amplification of the element.Translated from Genetika, Vol. 41, No. 1, 2005, pp. 23–29.Original Russian Text Copyright © 2005 by Kotnova, Karpova, Feoktistova, Lyubomirskaya, Kim, Ilyin.     

Defining Structural Domains of an Intrinsically Disordered Protein: Sic1, the Cyclin-Dependent Kinase Inhibitor of <Emphasis Type="Italic">Saccharomyces</Emphasis><Emphasis Type="Italic">cerevisiae</Emphasis>

The cyclin-dependent kinase inhibitor Sic1 is an intrinsically disordered protein (IDP) involved in cell–cycle regulation in the yeast Saccharomyces cerevisiae. Notwithstanding many studies on its biological function, structural characterization has been attempted only recently, fostering the development of production and purification protocols suitable to yield large amounts of this weakly expressed protein. In this study, we describe the identification of protein domains by the heterologous expression, purification, and characterization of Sic1-derived fragment. Four C-terminal fragments (Sic1^C-ter) were produced based on functional studies and limited-proteolysis results. The N-terminal fragment (Sic1^1–186) was complementary to the most stable C-terminal fragments (Sic1^Δ186). Both Sic1^1–186 and Sic1^C-ter fragments were, in general, less susceptible to spontaneous proteolysis than the full-length protein. The boundaries of the C-terminal fragments turned out to be crucial for integrity of the recombinant proteins and required two rounds of design and production. Sic1 fragments were purified by a simple procedure, based on their resistance to heat treatment, at the amount and purity required for structural characterization. Circular dichroism (CD) measurements and nuclear magnetic resonance (NMR) spectra of N- and C-terminal fragments confirm their disordered nature but reveal minor structural differences that may reflect their distinct functional roles.     

Limited functional conservation of a global regulator among related bacterial genera: Lrp in <Emphasis Type="Italic">Escherichia</Emphasis>, <Emphasis Type="Italic">Proteus</Emphasis> and <Emphasis Type="Italic">Vibrio</Emphasis>

Background  

Bacterial genome sequences are being determined rapidly, but few species are physiologically well characterized. Predicting regulation from genome sequences usually involves extrapolation from better-studied bacteria, using the hypothesis that a conserved regulator, conserved target gene, and predicted regulator-binding site in the target promoter imply conserved regulation between the two species. However many compared organisms are ecologically and physiologically diverse, and the limits of extrapolation have not been well tested. In E. coli K-12 the leucine-responsive regulatory protein (Lrp) affects expression of ~400 genes. Proteus mirabilis and Vibrio cholerae have highly-conserved lrp orthologs (98% and 92% identity to E. coli lrp). The functional equivalence of Lrp from these related species was assessed.     

New species of <Emphasis Type="Italic">Diphymyces</Emphasis> (Laboulbeniales,Ascomycota)

Four new species of Diphymyces parasitic on Leiodidae Cholevinae are described. These are D. arnaudii parasitic on Nemadiopsis spp. (Anemadini) from Chile, D. giachinoi parasitic on Nargiotes spp. (Anemadini) from Australia, D. pavicevicii parasitic on Pholeuonopsis magdelainei (Leptodirini) from Serbia, and D. pusillus parasitic on Adelopsis bioforestae (Ptomaphagini) from Ecuador.     

The use of HRMAS NMR spectroscopy to study the <Emphasis Type="Italic">in vivo</Emphasis> intra–cellular carbon/nitrogen ratio of <Emphasis Type="Italic">Solieria chordalis</Emphasis> (Rhodophyta)

The red alga Solieria chordalis (J. Agardh) C. Agardh (Rhodophyta) was used as a model to investigate the effects of changes in seawater salinity on the carbon/nitrogen ratio. Carbohydrates and nitrogenous metabolites are major components of this alga and their metabolisms are intimately linked. Previous publications have provided a solid database for these two primary metabolic pathways from experiments and observations in situ. Storage products (e.g. floridean starch), cell wall polysaccharides (carrageenan) and low molecular weight carbohydrates such as floridoside and digeneaside are major compounds constituting the pool of available carbon. Compounds such as amino acids and peptides, constitute the pool of nitrogen. This study focuses on the intracellular C/N ratio inside the pool of low molecular weight compounds. This C/N ratio can be defined as the balance between carbohydrates and amino acids. High–resolution magic angle spinning nuclear magnetic resonance spectroscopy (HRMAS NMR) provides a powerful approach for in vivo analysis of the pool of intracellular organic compounds. These in vivo results were complimented with quantitative data obtained from high performance liquid chromatography (HPLC). In vivo and in vitro experimental analyses provided a intracellular molecular balance and defined the C/N ratio. In order to study the effect of salt stress on the carbon/nitrogen ratio, S. chordalis was cultured under controlled conditions. Effects of hyposalinity and hypersalinity stresses (low 22 ‰ and high 50 ‰ salinity) were tested. Both HPLC and NMR data, obtained on stressed and unstressed algae, generated insights into variations of carbonated and nitrogenous metabolites, involving changes of the C/N ratios, and demonstrated the adaptive responses of the seaweed. 19th International Seaweed Symposium KOBE JAPAN.     

Isolation and characterization of antibiotics produced by <Emphasis Type="Italic">Streptomyces chrysomallus</Emphasis> R-21 and <Emphasis Type="Italic">S. globisporus</Emphasis> L-242 strains

Chrysomal and globerin complexes containing antibiotics from polyether and heptaene aromatic macrolide groups have been isolated from the mycelium of Streptomyces chrysomallus R-21 and Streptomyces globisporus L-242 strains, respectively. Physicochemical characteristics of these complexes have been investigated, including UV and NMR spectra, chromatographic parameters, etc. The fungicidal and antiviral activity of the studied complexes has been demonstrated on tomato plants. The obtained data allow us to consider the above-mentioned antibiotics to be original preparations.     

Examination of boldness traits in sexual and asexual mollies (<Emphasis Type="Italic">Poecilia latipinna</Emphasis>, <Emphasis Type="Italic">P. formosa</Emphasis>)

Considering the high costs of sexual reproduction (e.g., the production of males), its maintenance and predominance throughout the Animal Kingdom remain elusive. Especially the mechanisms allowing for a stable coexistence of closely related sexual and asexual species are still subject to a lively debate. Asexuals should rapidly outnumber sexuals due to higher population growth rates, unless they face some disadvantages. Here, we investigate potential differences in feeding behavior in a system of sexual (sailfin mollies, Poecilia latipinna) and coexisting gynogenetic fishes (Amazon mollies, Poecilia formosa). In two different experiments, we tested for differences in behavioral traits associated with boldness. Bold individuals take higher risks for gains in resources, so shyer individuals should be less competitive. Our study was motivated by the recent finding that P. formosa are less likely to be preyed upon by piscine predators than P. latipinna. We asked whether this result is indicative of low boldness in P. formosa. However, no differences between the two species were detectible in our behavioral experiments measuring (a) time to emerge from shelter to explore a novel environment, (b) latency time until feeding in a novel environment, and (c) recovery time until feeding restarted after a simulated predator attack. Furthermore, different boldness measures were not correlated with each other within individuals.     

The lens eyes of the box jellyfish <Emphasis Type="Italic">Tripedalia cystophora</Emphasis> and <Emphasis Type="Italic">Chiropsalmus sp.</Emphasis> are slow and color-blind

Box jellyfish, or cubomedusae, possess an impressive total of 24 eyes of four morphologically different types. Compared to other cnidarians they also have an elaborate behavioral repertoire, which for a large part seems to be visually guided. Two of the four types of cubomedusean eyes, called the upper and the lower lens eye, are camera type eyes with spherical fish-like lenses. Here we explore the electroretinograms of the lens eyes of the Caribbean species, Tripedalia cystophora, and the Australian species, Chiropsalmus sp. using suction electrodes. We show that the photoreceptors of the lens eyes of both species have dynamic ranges of about 3 log units and slow responses. The spectral sensitivity curves for all eyes peak in the blue-green region, but the lower lens eye of T. cystophora has a small additional peak in the near UV range. All spectral sensitivity curves agree well with the theoretical absorbance curve of a single opsin, strongly suggesting color-blind vision in box jellyfish with a single receptor type. A single opsin is supported by selective adaptation experiments.     

Seasonal changes in water source of four families of loblolly pine (<Emphasis Type="Italic">Pinus</Emphasis><Emphasis Type="Italic">taeda</Emphasis> L.)

Source water utilized by four families of loblolly pine (Pinus taeda L.) was assessed by comparing the H isotope composition ('D) of xylem sap and of soil water from four depths (0-20 cm, 20-40 cm, 1.2 m, and 2.1 m) across 1 year. Soil water 'D values varied with soil moisture content in the well-drained, sandy site and at each of the four soil depths. In September and November 1997 and May through November 1998, xylem sap 'D values closely matched the soil water 'D values of the upper soil horizons (0-20 and 0-40 cm) and, therefore, reflected significant water uptake from upper regions of the soil profile. In March 1998, xylem sap 'D values closely matched the soil water 'D values of the 1.2 m soil depth and, therefore, indicated that trees during this portion of the growing season were obtaining their water from deep in the soil profile. Analysis of source water use with a two-ended mixing model in the 3 months of collection that exhibited a range of soil water 'D values across the soil profile confirmed that trees utilized different sources of water depending upon season of the year. In September 1997 and November 1998, source water uptake was primarily from the upper soil profile while in March 1998, source water uptake was from deep in the soil profile. With few exceptions, we did not find striking differences in source water use between drought-hardy families and those that were locally adapted.     

Effects of site-directed mutagenesis of <Emphasis Type="Italic">mglA</Emphasis> on motility and swarming of <Emphasis Type="Italic">Myxococcus xanthus</Emphasis>

Background  

The mglA gene from the bacterium Myxococcus xanthus encodes a 22kDa protein related to the Ras superfamily of monomeric GTPases. MglA is required for the normal function of A-motility (adventurous), S-motility (social), fruiting body morphogenesis, and sporulation. MglA and its homologs differ from all eukaryotic and other prokaryotic GTPases because they have a threonine (Thr78) in place of the highly conserved aspartate residue of the consensus PM3 (phosphate-magnesium binding) region. To identify residues critical for MglA function or potential protein interactions, and explore the function of Thr78, the phenotypes of 18 mglA mutants were characterized.