Using protein synthesis inhibitors to establish the phylogenetic relationships of the sulfolobales order

The sensitivity of the cell-free protein synthesis systems from Acidanus brierleyi, Acidianus infernus, and Metallosphaera sedula, members of the archaeal order Sulfolobales, to 40 antibiotics with different specificities has been studied. The sensitivity patterns were compared to those of Sulfolobus solfataricus and other archaeal, bacterial, and eukaryotic systems. The comparative analysis shows that ribosomes from the sulfolobales are the most refractory to inhibitors of protein synthesis described so far. The sensitivity results have been used to ascertain in phylogenetic relationships among the members of the order Sulfolobales. The evolutionary significance of these results are analyzed in the context of the phylogenetic position of this group of extreme thermophilic microorganisms. Correspondence to: R. Amils     

Sound localisation in crickets

Intracellular recordings were made in the brain of the cricket Gryllus bimaculatus from an ascending auditory interneuron (AN1). Acoustic stimuli with calling song temporal pattern were delivered via earphones in a preparation with the acoustic trachea cut (attenuation of crossing sound > 30 dB). The input-output function of this cell was then determined by recording its responses to stimulation of the ipsilateral ear alone, of the contralateral ear alone and to stimulation of both ears simultaneously with the same or different carrier frequencies and intensities.This interneuron was excited by the ear ipsilateral to its axon and dendritic field and unresponsive to stimuli presented to the axon-contralateral ear alone. However, in binaural stimulation experiments, the response to a constant ipsilateral stimulus was progressively reduced as the intensity of a simultaneous contralateral stimulus was increased, above a threshold intensity.Tuning curves for threshold of this inhibition, determined in binaural stimulation experiments, indicated significant inhibition in the range 3–20 kHz with lowest threshold at 4–5 kHz. The inhibition was unaffected by sectioning of the contralateral circumoesophageal or neck connective, indicating that the inhibitory influence crosses the midline at the level of the prothoracic ganglion. Intracellular recordings from AN1 in the prothoracic ganglion confirmed that it was indeed neurally inhibited by inputs from the contralateral ear.Tuning curves for excitation of an omega neuron (ON1) by the ear ipsilateral to its soma and also the tuning of inhibition of ON1 by its contralateral ON1 partner, closely match the tuning of inhibition of AN1 and to a lesser extent, of AN2. This was taken as evidence that each AN1 is inhibited by the contralateral ON1. The significance of this interaction for directional hearing and phonotaxis is discussed.Abbreviations AP/CHP action potentials per chirp - AN1, AN2 ascending auditory interneurons 1, 2 - ON1 omega neuron 1 - ipsi ipsilateral contra contralateral - PTG prothoracic ganglion loc lateral ocellar nerve - On optic nerve an antennal nerve - coc circum-oesophageal connective so sound off     

Cyanobacteria of Rocks and Soils of the Orinoco Lowlands and the Guayana Uplands,Venezuela

The occurrence of 23 cyanobacterial species, belonging to 9 different genera and 5 cyanobacterial lichen species of 5 different genera on exposed, open rock surfaces of inselbergs and on soil in savannas of the Orinoco lowlands and the Guayana uplands is described. Their distribution patterns and frequency within the different habitats are given. The filamentous procaryotic blue-green algae/cyanobacteria Stigonema ocellatum and Scytonema crassum, together with the unicellular cyanobacterium Gloeocapsa sanguinea were the most frequent species on rocks, whereas the filamentous cyanobacterium, Schizothrix telephoroides, dominated in cyanobacterial mats on the savanna soil. All species showed intensively coloured sheaths, either brown or yellow in the case of Stigonema ocellatum and Scytonema crassum, or red in Gloeocapsa sanguinea and Schizothrix telephoroides. In addition, a number of cyanobacterial lichens occurred.     

Male-male pairs in Greylag Geese (Anser anser)

Summary For almost two decades a flock of 130 free-flying Greylag Geese (Anser anser) has been the focus of detailed ethological investigations at the Konrad Lorenz Institut in Grünau im Almtal, Austria. Gander pairs, i. e. male-male pairs, represent a prominent social unit in this flock and were the subject of a detailed behavioral investigation. Analysis of the composition and dynamics of the flock over a 15 year period indicated that the incidence of homosexual pairings closely paralleled the male bias of the sex ratio. The behavior of ganders in gander pairs was investigated and compared to that of gander and goose in heterosexual pairs. The behavior of the two males in a gander pair (1) was comparable in most aspects, (2) was similar to the behavior of the gander in heterosexual pairs, and (3) differed greatly from that of the heterosexually paired goose. Therefore, pseudo-female behavior in one partner cannot account for the formation of a pairbond between two males. As a unit, gander pairs were characterized by a higher frequency of offensive agonistic behavior compared to heterosexual pairs and spent significantly more time peripheral to, and away from the flock than did heterosexual pairs.

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Zusammenfassung Das Sozialgefüge einer Schar Graugänse ist weitaus komplizierter, als es das monogame Fortpflanzungssystem erwarten ließe (Collias &Jahn 1959,Fischer 1965,Kalas 1979,Rutschke 1982). Ganterpaare, die häufig über Jahre hinweg bestehen bleiben, sind für tiersoziologische Untersuchungen interessant, weil ihre Funktion nicht im Rahmen der Fortpflanzung gesehen werden kann. Welche Bedingung begünstigen die Bildung von Ganterpaaren, und welche Verhaltensmechanismen tragen zum Entstehen und zur Aufrechterhaltung dieser Verbindung bei? Die Zusammensetzung der Grünauer Graugansschar 1973–1988 zeigt, daß die Anzahl der Ganterpaare von einem Überschuß von Männchen in der Schar abhängt. Das Verhalten von 6 Ganterpaaren wurde untersucht und mit dem von heterosexuellen Paaren verglichen. Innerhalb eines Ganterpaares entsprachen sich die Partner in der Häufigkeit von agonistischem sowie sozial-bindendem Verhalten. Homo- und heterosexuell verpaarte Ganter zeigten sich im Verhalten vergleichbar. Der Ganter eines Ganterpaares unterschied sich jedoch in der Häufigkeit aller untersuchten Verhaltensweisen von dem der heterosexuell verpaarten Gans. Folgende Schlußfolgerungen und Hypothesen bieten sich an: (1) Pseudo-weibliches Verhalten bei einem der Ganter scheint nicht die Bildung von Ganterpaaren erklären zu können. Beide Ganter verhalten sich rein männlich und behandeln den Partner so, als ob dieser ein Weibchen wäre. (2) Ein Mangel an gegengeschlechtlichen Schargenossen fördert die Bildung von homosexuellen Paaren und aufzuchtsbekannte Vögel werden dabei vorgezogen. (3) Ein Zusammenschluß mit einem gleichgeschlechtlichen Artgenossen sollte, verglichen mit der Möglichkeit alleine zu bleiben, eine überlegene Strategie darstellen, da unverpaarte Gänse geringeren Zugang zu Futterquellen haben und eher Raubtieren zum Opfer fallen. (4) Homosexuelle Paare könnten als ein Puffersystem für Ganter angesehen werden, vor allem zu Zeiten in denen das Geschlechtsverhältnis in Richtung der Männchen verschoben ist. (5) Aggression des Ganters richtet sich generell gegen andere männliche Schargenossen. Die Bildung von Ganterpaaren, also besonders aggressiven Paaren, könnte daher dazu beitragen, Ganter aus der Schar zu vertreiben und ein Übermaß von Männchen in der Schar zu verhindern. (6) Da wir zeigen konnten, daß sich homosexuelle Paare oft am Rande der Schar aufhalten und dabei häufig sichern, könnte solchen Paaren eine Art Wächterfunktion zukommen. (7) Andererseits ist es durchaus möglich, daß Ganterpaare bloß ein Epiphenomen einer Graugansschar mit einem Überschuß an Männchen darstellen.

     

Detection of enteroviruses in groundwater with the polymerase chain reaction.

Standard methods for the detection of enteroviruses in environmental samples involve the use of cell culture, which is expensive and time-consuming. The polymerase chain reaction (PCR) is an attractive method for the detection of enteroviruses in water because primary cell culture is not needed and the increased sensitivity of PCR allows detection of the low numbers of target DNAs and RNAs usually found in environmental samples. However, environmental samples often contain substances that inhibit PCR amplification of target DNA and RNA. Procedures that remove substances that interfere with the amplification process need to be developed if PCR is to be successfully applied to environmental samples. An RNA-PCR assay for the detection of enteroviruses in water was developed and used to test a variety of groundwater concentrates and humic acid solutions seeded with poliovirus type 1. The groundwater samples and humic acid solutions were treated with Sephadex G-50, Sephadex G-100, Sephadex G-200, Chelex-100 resin, and a mixed bed resin to remove PCR-inhibitory material from the samples. Sephadex G-100 in combination with Chelex-100 was found to be very effective in removing inhibitory factors for the detection of enteroviruses in groundwater concentrates by PCR. Viruses were detected in two of the groundwater concentrates by the RNA-PCR assay after treatment with Sephadex G-100 plus Chelex-100. This was confirmed by tissue culture, suggesting that the treatment protocol and, subsequently, the RNA-PCR assay are applicable for the detection of enteroviruses in environmental samples.     

Improved mating efficiency inAcetabularia acetabulum: recovery of 48–100% of the expected zygotes

Summary We have improved zygote recovery 11–1,000 fold by optimizing the physiology of gamete release and mating inAcetabularia acetabulum. Gamete release was affected by agar purity, concentration, and volume/gametangial pair. Cold pre-treatment of gametangia (14–30 d at 10°C in the dark) synchronized subsequent gamete release at 21°C in the light. Cold pre-treatment was nearly twice as effective in synchronizing subsequent gamete release when intact, gametangia-bearing caps rather than isolated gametangia were pretreated. Synchronizing gamete release doubled mating efficiency. In a wild-type laboratory strain ofA. acetabulum, there were 1,561±207 gametes/gametangium which had half-lives of 14.5 d in 0.1% seawater-agar. We recovered 48–93% of the expected numbers of zygotes from a mass mating of 8 to 1,226 gametangia and 11–128% of the expected numbers of zygotes from mating single gametangial pairs: the large range in the calculated mating efficiency may be attributable to the variation in the numbers of gametes made per gametangium. Zygote recovery from single gametangial pairs was highly dependent on the volume of mating matrix. In addition, most zygotes recovered were unattached to any other zygotes in the subsequent generation (> 95% single cells from matings of 1–500 gametangial pairs). Our improvements in mating conditions and zygote recovery (1) have facilitated cell manipulation and culture ofA. acetabulum in the laboratory; and (2) have made controlled crosses for selection and genetic analysis of mutants feasible. These advances have removed a major barrier to genetic analysis of development inAcetabularia.Abbreviations LB Luria-Bertani bacteriological broth - SE standard error of the mean - T_g agar gelling temperatures - DAPI 4,6-diamidino-2-phenylindole     

Small-spored Alternaria spp. (section Alternaria) are common pathogens on wild tomato species

The wild relatives of modern tomato crops are native to South America. These plants occur in habitats as different as the Andes and the Atacama Desert and are, to some degree, all susceptible to fungal pathogens of the genus Alternaria. Alternaria is a large genus. On tomatoes, several species cause early blight, leaf spots and other diseases. We collected Alternaria-like infection lesions from the leaves of eight wild tomato species from Chile and Peru. Using molecular barcoding markers, we characterized the pathogens. The infection lesions were caused predominantly by small-spored species of Alternaria of the section Alternaria, like A. alternata, but also by Stemphylium spp., Alternaria spp. from the section Ulocladioides and other related species. Morphological observations and an infection assay confirmed this. Comparative genetic diversity analyses show a larger diversity in this wild system than in studies of cultivated Solanum species. As A. alternata has been reported to be an increasing problem in cultivated tomatoes, investigating the evolutionary potential of this pathogen is not only interesting to scientists studying wild plant pathosystems. It could also inform crop protection and breeding programs to be aware of potential epidemics caused by species still confined to South America.     

Crystal structure of human annexin I at 2.5 A resolution.

cDNA coding for N-terminally truncated human annexin I, a member of the family of Ca(2+)-dependent phospholipid binding proteins, has been cloned and expressed in Escherichia coli. The expressed protein is biologically active, and has been purified and crystallized in space group P2(1)2(1)2(1) with cell dimensions a = 139.36 A, b = 67.50 A, and c = 42.11 A. The crystal structure has been determined by molecular replacement at 3.0 A resolution using the annexin V core structure as the search model. The average backbone deviation between these two structures is 2.34 A. The structure has been refined to an R-factor of 17.7% at 2.5 A resolution. Six calcium sites have been identified in the annexin I structure. Each is located in the loop region of the helix-loop-helix motif. Two of the six calcium sites in annexin I are not occupied in the annexin V structure. The superpositions of the corresponding loop regions in the four domains show that the calcium binding loops in annexin I can be divided into two classes: type II and type III. Both classes are different from the well-known EF-hand motif (type I).     

Diol sulfonamides: A potent and novel class of inhibitors of human renin

The syntheses and pharmacological activity of a series of diol sulfonamides which function as inhibitors of human renin are described. The most potent compound in this series, compound 20 (SQ 33,800), is a subnanomolar inhibitor of human renin (IC₅₀ = 0.35 × 10⁻⁹ M).