Behavioral Ecology of Elliot’s Laughingthrush (<Emphasis Type="Italic">Trochalopteron</Emphasis> (<Emphasis Type="Italic">Garrulax</Emphasis>) <Emphasis Type="Italic">elliotii</Emphasis>, Timaliidae,Aves): 2. Vocal Repertoire

Our work is the first study on the vocal repertoire of one of the babbler species, Elliot’s laughingthrush. Field studies were carried out in Hupingshan Nature Reserve, Hunan Province, China. There are three types of signals in the repertoire: songs, calls (chattering), and duets. Songs and calls are used by both males and females. The song consists of a quiet introduction (short note) and a louder main part (two or three tonal notes). There are up to four song variants in the repertoire of a given pair. All songs can be classified into three types. Types I and II represent songs with the main part consisting of three notes, while type III consists of two notes. Types I and II differ from each other in certain features of the frequency modulation shape of the first two notes. All song types can be used during the spontaneous vocalization of a single bird. However, their usage in other contexts differs. Songs of types I and II are more often uttered during vocal interactions of neighbor males. Songs of type III appear to be characteristic of male–female duets. Chattering (calls) is a continuous series of broadband notes. There are several (two to five) note variants in each series. Chattering can be used both during a male–female interaction and as an alarm call. We found an inverse correlation between (1) the duration of pauses between notes, and (2) the number of note variants (“repertoire size”) in a series. Both parameters probably reflect the internal state of an individual at a given moment.     

Revision of “<Emphasis Type=Italic>Septonema ochraceum</Emphasis>” revealed three new species of <Emphasis Type=Italic>Venturiaceae</Emphasis> and <Emphasis Type=Italic>Herpotrichiellaceae</Emphasis>

Survey of seven strains determined as Septonema ochraceum (Dothideomycetes, inc. sed.) isolated from pine litter or obtained from public collections revealed three new species, Fusicladium cordae, F. sicilianum (Venturiaceae), Cladophialophora matsushimae (Herpotrichiellaceae) and a cryptic species morphologically identical to Devriesia americana (Teratosphaeriaceae), but phylogenetically distinct. Morphological survey and phylogenetic analysis using nucleotide sequence data from the nuclear ribosomal subunit genes indicate a close relationship within three species colonising pine litter needles, F. cordae, F. pini and F. ramoconidii. F. sicilianum is most related to F. rhodense. C. matsushimae represents a species belonging to one of the lineages of the polyphyletic genus Cladophialophora. None of the strains observed can be classified morphologically as S. ochraceum, of which the type material does not exist.     

Small colony variants of <Emphasis Type=Italic>Staphylococcus aureus</Emphasis> — <Emphasis Type=Italic>review</Emphasis>

Bacterial variants of Staphylococcus aureus called small colony variants (SCVs) originate by mutations in metabolic genes, resulting in emergence of auxotrophic bacterial subpopulations. These variants are not particularly virulent but are able to persist viable inside host cells. SCVs show their characteristic auxotrophic growth deficiency and depressed α-cytotoxin activity. Environmental pressure such as antibiotics, select for isogenic SCV cells that are frequently found coexisting with their parent wild-type strains in a mixed bacterial culture. SCV strains often grow on blood agar as non-pigmented or pinpoint pigmented colonies and their key biochemical tests are often non-reactive. Their altered metabolism or auxotrophism can result in long generation time and thus SCV phenotype, more often than not SCV can be overgrown by their wild-type counterparts and other competitive respiratory flora. This could affect laboratory detection. Thus, molecular methods, such as 16S rRNA partial sequencing or amplification of species-specific DNA targets (e.g. coagulase, nuclease) directly from clinical material or isolated bacterial colonies, become the method of choice. Patients at risk of infection by S. aureus SCVs include cystic fibrosis patients (CF), patients with skin and foreign-body related infections and osteomyelitis, as they suffer from chronic staphylococcal infections and are subject to long-term antibiotic therapy. Molecular evidence of SCV development has not been found except for some random mutations of the thymidylate synthase gene (thyA) described in SCV S. aureus strains of CF patients. These variants are able to bypass the antibiotic effect of folic acid antagonists such as sulfonamides and trimethoprim. Resistance to gentamicin and aminoglycosides in the hemin or menadione auxotrophic SCVs was hypothesized as being due to decreased influx of the drugs into cells as a result of decreased ATP production and decreased electrochemical gradient on cell membranes.     

Decolourisation and dephenolisation potential of selected <Emphasis Type=Italic>Aspergillus</Emphasis> section <Emphasis Type=Italic>Nigri</Emphasis> strains – <Emphasis Type=Italic>Aspergillus tubingensis</Emphasis> in olive mill wastewater

Aspergillus section Nigri strains Aspergillus aculeatus Ege-K 258, A. foeditus var. pallidus Ege-K156, A. niger Ege-K 4 and A. tubingensis Ege-K 265 were used to treat olive mill wastewater (OMW) in an investigation aimed at exploring their dephenolisation and decolourisation ability and, consequently, the economic feasibility of using any or all of these strains in a pre-treatment step in the processing of OMW. Of these strains A. tubingensis Ege-K 265 resulted in an 80% decolourisation of twofold-diluted OMW and a 30% decolourisation of undiluted OMW; in addition, it was able to remove approximately 30% of all phenolic compounds in both twofold-diluted and undiluted OMW. We conclude that A. tubingensis Ege-K 265 could be effectively used in the pre-treatment step of a combined aerobic-anaerobic process to solve the environmental problems caused by OMW in Mediterranean countries.     

Synthesis and use of <Emphasis Type=Italic>N</Emphasis>,<Emphasis Type=Italic>N</Emphasis>-di-Boc-glutamate γ-semialdehydes and related aldehydes

Summary.  This review article focuses on the synthesis and reactions of N,N-di-Boc glutamate and aspartate semialdehydes as well as related aldehydes. These building blocks are prepared according to various strategies from glutamic and aspartic acids and find interesting synthetic applications. In the first part, the methods for the synthesis of N,N-di-Boc-amino aldehydes are summarized. The applications of these chiral synthons for the synthesis of unnatural amino acids and other bioactive compounds are discussed in the second section. Received April 24, 2002 Accepted August 13, 2002 Published online January 30, 2003 Authors' address: Prof. Violetta Constantinou-Kokotou, Chemical Laboratories, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece, E-mail: vikon@aua.gr Abbreviations: AcNH-TEMPO, 4-acetamido-2,2,6,6-tetramethyl-1-piperidinyloxy free radical; AIBN, 2,2′-azobis(2-methylpropionitrile); Aliquat, methyltrioctylammonium chloride; Bn, benzyl; Boc, tert-butoxycarbonyl; Bu^t, tert-butyl; m-CPBA, 3-chloroperoxybenzoic acid; DAST, diethylaminosulfur trifluoride; DBU, 1,8-diazabicyclo[5.4.0]undec-7-ene; (R,R)-(+)-DET, (R,R)-(+)-diethyltartrate; DIBALH, diisobutyl aluminium hydride; DMAP, 4-dimethylaminopyridine; DMF, dimethylformamide; Et₃N, triethylamine; KHMDS, potassium bis(trimethylsilyl)amide; (S)-LLB, lanthanium-lithium-bis-metallic binaphthol catalyst; MsCl, methanesulfonyl chloride; NEM, N-ethylmorpholine; NMO, 4-methylmorpholine N-oxide; PPA, propylphosphonic acid anhydride; TBHP, tert-butyl hydroperoxide; TFA, trifluoroacetic acid; THF, tetrahydrofuran; TMSI, 1-(trimethylsilyl)imidazole; Trt, trityl.     

<Emphasis Type=Italic>Cyperaceae Raddianae</Emphasis>: a nomenclatural and taxonomic study of the Cyperaceae published in G. Raddi’s <Emphasis Type=Italic>Agrostografia brasiliensis</Emphasis>

A study of the 26 Cyperaceae published in G. Raddi’s Agrostografia brasiliensis is presented. Several new synonyms and a re-evaluation of some taxa described by G. Raddi are proposed. The first citation of Fimbristylis pilosa Vahl for Brazil is reported. The new combination, Pleurostachys pilulifera (Bertol.) Longhi-Wagner, Baldini & A. C. Araújo is made.     

Antimutagenic effect of lectin and <Emphasis Type=Italic>N</Emphasis>-methyl-<Emphasis Type=Italic>N’</Emphasis>-nitro-<Emphasis Type=Italic>N</Emphasis>-nitrosoguanidine on induced mutagenesis in mammalian cells in vitro

Factors and ways in which macromolecules influence the mutation process are considered. An antimutagenic effect is demonstrated in a study of the combined influence of lectins and the alkylating agent N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) on mutagenesis in Chinese hamster cells. Thus, in different schemes of treatment of cells with albumin and MNNG, the experimental frequency of mutations induced by the two agents was statistically reliably lower than the theoretically expected level for their independent combined action. The possibility that there exist common targets and mechanisms through which different types of mutagenic agents act on the cell DNA is discussed.     

A monograph of <Emphasis Type=Italic>Octoknema</Emphasis> (Octoknemaceae — Olacaceae <Emphasis Type=Italic>s.l.</Emphasis>)

A revision of Octoknema Pierre is provided, based on morphological data gathered from a study of herbarium specimens and observations in the field. Fourteen species of Octoknema are recognised including six new species: O. bakossiensis Gosline & Malécot, O. belingensis Gosline & Malécot, O. chailluensis Malécot & Gosline, O. kivuensis Gosline & Malécot, O. mokoko Gosline & Malécot and O. ogoouensis Malécot & Gosline. Data are given for four additional poorly known taxa (Octoknema species A, B, C and D).     

High-level Expression of Maize <Emphasis Type=Italic>γ</Emphasis>-zein Protein in Transgenic Soybean (<Emphasis Type=Italic>Glycine max</Emphasis>)

Primers were developed for 118 microsatellites isolated from grape (Vitis vinifera) genomic libraries enriched for (AC)n repeats. Only one microsatellite sequence matched other grape SSR-sequences in the GeneBank database. Genotyping was carried out in the parental lines and four offspring of two pseudo-test-cross populations, Cabernet Sauvignon x Seyval and Chardonnay x Bianca, and a further six other grape genotypes (V. vinifera Sultanina, Merlot, Syrah, Müller-Thurgau, Vitis Regent and V. riparia Gloire de Montpellier). A total of 108 microsatellites showed easily scorable alleles and 100 of them segregated according to a configuration suitable for mapping in either cross. A further 8 SSRs, although unsuitable for mapping in those crosses, showed polymorphism in the other genotypes tested. This set of markers was used, along with 75 microsatellites of other repeat-types, to fingerprint 46 offspring of the cross Chardonnay x Bianca. For each full-sib, individual heterozygosity and distance in repeat units between pairs of alleles at each locus (mean d²) were calculated as a tool for predicting highly outbred recombinant individuals. Six microsatellites with segregation ratios significantly distorted towards the lack of homozygous sibs were identified and mapped to linkage groups LG 3 and LG 5. Estimation of heterozygosity at genome-wide level and genotyping at loci for which homozygous sibs are discriminated against are discussed for marker-assisted background selection in outcrossing grapevines.     

Proteolysis of <Emphasis Type=Italic>α</Emphasis>-amylase from <Emphasis Type=Italic>Saccharomycopsis fibuligera</Emphasis>: characterization of digestion products

α-Amylase from Saccharomycopsis fibuligera R-64 was successfully purified by butyl Toyopearl hydrophobic interaction chromatography, followed by Sephadex G-25 size exclusion and DEAE Toyopearl anion exchange chromatography. The enzyme has a molecular mass of 54 kDa, as judged by SDS PAGE analysis. Upon tryptic digestion, two major fragments with relative molecular masses of 39 kDa and 10 kDa, which resemble the A/B and C-terminal domains in the homologous Taka-amylase, were obtained and were successfully separated with the Sephadex G-50 size exclusion column. The 39-kDa fragment demonstrated a similar amylolytic activity to that of the undigested enzyme. However, it was found that the K _m value of the 39-kDa fragment was about two-times higher than that of the undigested enzyme. Moreover, thermostability studies showed a lower half-life time for the 39-kDa fragment. These findings suggest that the 39-kDa fragment is the catalytic domain, while the 10-kDa fragment is the C-terminal one, which plays a role in thermostability and starch binding. Although the undigested enzyme is able to act on raw starches at room temperature, with maize starches as the best substrate, neither the undigested enzyme nor the fragments adsorb the tested raw starches. These results propose Saccharomycopsis fibuligera α-amylase as a raw starch-digesting but not adsorbing amylase, with a similar domain organization to that of Taka-amylase A.     

Botanical-geographical and forestry aspects of introgressive hybridization of the Gmelin’s larch (<Emphasis Type=Italic>Larix gmelinii</Emphasis> (Rupr.) Rupr.) and Cajander larch (<Emphasis Type=Italic>L. cajanderi</Emphasis> Mayr)

The paper studies the introgressive hybridization between the Gmelin’s larch and Cajander larch. The obtained data confirm the indisputable specific independence of the Cajander larch, which occupies about 48% of the total area of the larch forests in Russia. Its specific independence has been doubted by some experts. Once again more reliable and vast material shows the presence of wide transitional belt of hybrid populations at the juncture of the hybridizing species. Some forestry characteristics of the Gmelin’s larch and Cajanderi larch are studied that play important role in their natural restoration.     

NaCl-induced photoinhibition and recovery of the photosynthetic activity of a <Emphasis Type=Italic>katG</Emphasis><Superscript>−</Superscript> mutant of cyanobacterium <Emphasis Type=Italic>Synechocystis</Emphasis> sp. PCC 6803

The joint effects of 0.5 M NaCl and light of different intensities on the activity of the photosynthetic apparatus and ATP content in cells of the katG ⁻ mutant of cyanobacterium Synechocystis sp. PCC 6803 have been studied. The mutant demonstrated a higher photoinhibition rate and a slower rate of recovery compared with the wild type, as shown by measurements of the CO₂-dependent O₂ production and delayed fluorescence of Chl a. The presence of 0.5 M NaCl in the incubation medium caused equal photoinhibition of the photosynthetic apparatus at I = 1200 μE m⁻² s⁻¹ in the mutant and wild-type cells. At I = 2400 μE m⁻² s⁻¹, we observed stronger inhibition and slower recovery of the photosynthetic apparatus in the katG ⁻ mutant than in wild-type cells. The data obtained evidence an important role of catalase-peroxidase in the system of reparation of the photosynthetic apparatus damaged by high-intensity light, especially at the background of NaCl stress.     

High-level expression,purification and characterization of a recombinant medium-temperature <Emphasis Type=Italic>α</Emphasis>-amylase from <Emphasis Type=Italic>Bacillus subtilis</Emphasis>

Alpha-amylases are important industrial enzymes with a wide range of applications. Although medium-temperature alpha amylase (AmyE) has some practical advantages, its low yield has limited its applications. When an amyE gene from Bacillus subtilis BF768 was cloned into vector pWB980 and over-expressed in B. subtilis WB600, high activities (723 U ml⁻¹) of secreted AmyE were produced. Recombinant AmyE was purified to a specific activity of 36 U mg⁻¹ having optimal activity at pH 6.0 and 60°C.