Computerized topographical analysis of functionally homogeneous neuronal sets.

A computer-assisted analysis of the spatial distribution of neurons having homogeneous characteristics is described in this paper. The camera lucida drawings of sections of a brain nucleus and the points representing the neurons labeled on the basis of a specific behavior of discharge rates were digitized on a personal computer Amiga 2000 or IBM compatible. Our software provided: a) the computerized, stereotaxically oriented reconstruction of the stored sections and of the plotted neurons; b) the identification within each section of the mass center (MC) of the units sharing a given behavior and of the area where the density of such neurons was maximal (MDA). The routine was tested on the spatial distribution of neuronal responses to serotonin in the lateral vestibular nucleus.     

Inbreeding reveals interpopulation differences in inversion polymorphism of Drosophila subobscura

The adaptive significance of inversion polymorphism of Drosophila subobscura is well established. However, little is known about gene combinations within inversions which are coadapted because of population-specific effects. We studied this aspect of Dobzhansky's coadaptation hypothesis, using the systematic inbreeding method. Differences in magnitude and quality of inversion polymorphism in two ecologically and topologically distinct habitats were compared after several generations of continuous full-sib inbreeding. Populations from the two habitats differ in frequency of homokaryotypes after third and fifth generation of inbreeding and in the levels of homozygosity of different gene arrangements. The effect of homozygosity appears population and chromosome specific.     

Carotenoid synthesis in Streptomyces setonii ISP5395 is induced by the gene crtS, whose product is similar to a sigma factor

In many species of actinomycetes, carotenogenesis can be photoinduced. The capacity to respond to photoinduction is, however unstable and, in various strains of Streptomyces, is lost at a relatively high frequency. In Streptomyces setonii ISP5395, which normally produces no carotenoids, carotenoid-producing mutants can be obtained following protoplast regeneration. We report here the characterization of a gene, crtS, which was isolated from one such mutant and can confer on wild-type S. setonii ISP5395 cells the capacity to synthesize carotenoids. Sequence analysis of crtS reveals an open reading frame, which shows homology to genes that encode alternative sigma factors in Bacillus subtilis. We propose that crtS encodes a sigma factor which is necessary for the expression of a cryptic gene(s) for carotenoid biosynthesis in S. setonii ISP5395.     

Assay of 1L-myo-inositol-1-phosphatase using a fluorometric method.

1L-myo-Inositol-1-phosphatase, an enzyme purified from brain tissues, catalyzes the dephosphorylation of 1L-myo-inositol 1-phosphate. This enzyme has become the subject of intense research interest, since myo-inositol is needed for the resynthesis of phosphatidylinositol. We have developed a sensitive fluorometric assay for detecting the activity of 1L-myo-inositol-1-phosphatase. The assay is based on o-aminobenzoyl beta-glycerophosphate fluorescence, according to the following principles: (I) The fluorescence yield of o-aminobenzoyl beta-glycerophosphate is increased by 2.75-fold in the presence of saturating concentrations of bovine serum albumin. (II) o-Aminobenzoyl beta-glycerophosphate has the same fluorescence yield as o-aminobenzoyl glycerol, but the latter does not bind to bovine serum albumin. (III) Dephosphorylation of the substrate, catalyzed by the monophosphatase, makes less o-aminobenzoyl beta-glycerophosphate available for binding to bovine serum albumin, thereby producing a decrease in the fluorescence intensity.     

Vegetation fluctuation in mediterranean dune ponds in relation to rainfall variation and water extraction

Abstract. The structure of dune ponds hygrophytic vegetation has a spatial organisation in belts around the pond basin, closely related to water level and flooding regime. Doñana National Park has an important representation of temporal dune ponds, which are subjected to rainfall fluctuations and may be suffering the impact of water extraction from the neighbouring tourist resort. Permanent transects in a vegetation complex of five dune ponds have been monitored during a eight year period (1990-1997). This period was characterised by a number of dry years (annual rainfall around 300 mm), located between two wet cycles (800-900 mm). Transects were analysed in 1990 (wet period), 1994 (dry) and 1997 (wet) by hierarchical agglomera-tive clustering. During the dry period hygrophytic species showed regression, with a high mortality of some species like Ulex minor, while the xerophytic species advanced to lower areas. Seedlings of some xerophytic species colonised the dry surface of the pond basin. The lowering of the water table varied in the different ponds, ranging from 312 to 190 cm depending on topography and the distance to the pumping area. The new period of flooding during 1995-96 and 1996-97 cycles provided the opportunity for hygrophytic spe cies to re-establish themselves in their original places. This study suggest that changes in vegetation are caused by the interaction between weather conditions and human disturbance (water extractions). In our example man-made disturbance is more marked during the dry periods while wet periods tend to obscure the effects of water extractions. From a management perspective, long-term monitoring of water table and vegetation structure is revealed as a key procedure to the management of land-water ecotones on pressured areas and threatened habitats.     

Transposition-mediated transcriptional overexpression as a mechanism of insecticide resistance

It has been proposed that amplification of genes for esterase that provide resistance to insecticides may originate from transposition events. To test this hypothesis, we have constructed a minigene coding for a soluble acetylcholinesterase under the control of a nontissue-specific promoter (hsp70). When introduced into Drosophila, the gene is expressed in all tissues and the extra acetylcholinesterase produced confers a low level of insecticide resistance (twofold). The minigene was mobilized by crossing the initial transformant with a strain providing a source of P-element transposase. After 34 generations of exposure to the organophosphate parathion, we obtained a strain with a higher resistance (fivefold). This strain had only one extra Ace gene, which overexpressed acetylcholinesterase. Thus, following transposition, resistance resulted from the overexpression of a single copy of the gene and not from gene amplification. Received: 9 August 1996 / Accepted: 27 May 1997     

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Transient scrotal hyperthermia and levonorgestrel enhance testosterone-induced spermatogenesis suppression in men through increased germ cell apoptosis.     

Cathepsin B from human renal cortex.

Cysteine-proteinase activity was observed in homogenates of human-cadaver renal cortex. This activity co-purified with renin enzymic activity until separation by aminohexyl-Sepharose--pepstatin affinity chromatography. The cysteine proteinase was purified 1780-fold after the following successive chromatographic procedures: Sephadex G-75, DEAE-cellulose DE-52, and an organomercurial affinity resin. The proteinase activity was dependent upon activation by thiol-containing compounds such as dithiothreitol, as well as by EDTA, and was inhibited by the thiol-group-specific alkylating reagents iodoacetic acid and N-ethylmaleimide. DE-52 cellulose chromatography resolved the cysteine proteinase into two components. On the basis of molecular size (26 000 daltons), activity as a function of pH, stability as a function of pH, substrate specificity and thermal lability, the major component (95%) has been identified as cathepsin B. The DE-52 cellulose elution pattern of the minor component (5%) is suggestive of cathepsin H [Schwartz & Barrett (1980) Biochem. J. 191, 487-497] Enzymic activity was determined with synthetic substrates, in particular alpha-N-benzoyl-DL-arginine 2-naphthylamide (Bz-Arg-NNap), thus precluding the detection of cathepsin L [Kirschke, Langner, Wiederanders, Ansorge, Bohley & Broghammer (1976) Acta Biol. Med. Germ. 35, 285-299]. Inhibition by dimethyl sulphoxide was observed in the determination of Km = 7.0 +/- 0.4 mM for the substrate Bz-Arg-NNap, and care must therefore be taken in the preparation of substrate solutions.