Changes in Race-Specific Virulence in Pseudomonas syringae pv. phaseolicola Are Associated with a Chimeric Transposable Element and Rare Deletion Events in a Plasmid-Borne Pathogenicity Island

Virulence for bean and soybean is determined by effector genes in a plasmid-borne pathogenicity island (PAI) in race 7 strain 1449B of Pseudomonas syringae pv. phaseolicola. One of the effector genes, avrPphF, confers either pathogenicity, virulence, or avirulence depending on the plant host and is absent from races 2, 3, 4, 6, and 8 of this pathogen. Analysis of cosmid clones and comparison of DNA sequences showed that the absence of avrPphF from strain 1448A is due to deletion of a continuous 9.5-kb fragment. The remainder of the PAI is well conserved in strains 1448A and 1449B. The left junction of the deleted region consists of a chimeric transposable element generated from the fusion of homologs of IS1492 from Pseudomonas putida and IS1090 from Ralstonia eutropha. The borders of the deletion were conserved in 66 P. syringae pv. phaseolicola strains isolated in different countries and representing the five races lacking avrPphF. However, six strains isolated in Spain had a 10.5-kb deletion that extended 1 kb further from the right junction. The perfect conservation of the 28-nucleotide right repeat of the IS1090 homolog in the two deletion types and in the other 47 insertions of the IS1090 homolog in the 1448A genome strongly suggests that the avrPphF deletions were mediated by the activity of the chimeric mobile element. Our data strongly support a clonal origin for the races of P. syringae pv. phaseolicola lacking avrPphF.     

Outdoor pilot-scale production of Botryococcus braunii in panel reactors

In this paper, the outdoor production of Botryococcus braunii in pilot-scale panel reactors (0.4?m³) is studied under uncontrolled conditions at a location close to the Atacama Desert (Chile). Discontinuous experiments were performed on different dates to determine the feasibility of the culture and the influence of environmental conditions on the system yield. Data showed that solar radiation is a major parameter in determining system yield, the average irradiance inside the culture determining both the growth rate and biomass productivity. A maximum specific growth rate of 0.09?day^?1 and biomass productivity of 0.02?g?L^?1?day^?1 (dry weight) were measured in discontinuous mode, at an average irradiance of 60?μE?m^?2?s^?1. With respect to lipids, a productivity of 2.5?mg?L^?1?day^?1 was obtained under favourable growth conditions; no accumulation of lipids at the stationary phase was observed. To confirm this behaviour, a semicontinuous culture was performed at 0.04?day^?1 in a larger reactor (1?m³). In this experiment, the biomass concentration and productivity was 0.3?g?L^?1 and 0.015?g?L^?1?day^?1, respectively. The lipid content and productivity was 15.6% and 2.4?mg?L^?1?day^?1, respectively, the mean average irradiance inside the reactor being 60?μmol photons?m^?2?s^?1. The light path of the reactor determines the light availability, thus determining also the biomass concentration and productivity of the reactor once the dilution rate is fixed. Experimentally, biomass productivity of 0.015?g?L^?1?day^?1 was determined for a light path of 0.15?m, but this can be increased by more than three times for a light path of 0.1?m. These data confirm that this alga can be produced outdoors in a secure form, the culture yield improving when optimal conditions are applied, the data reported here establishing the starting point for the development of the process.     

Close Encounters of the First Kind: Innate Sensors and Multiple Sclerosis

Although autoimmune diseases by definition imply adaptive immune system pathologies, growing evidence points to the relevance of innate receptors in modulating the initiation and progression of the autoreactive response. Multiple sclerosis (MS) is a chronic autoimmune disease characterised by central nervous system (CNS) demyelination, inflammation and axonal damage, in which the role of several pathogens such as herpes viruses have long been described as potential triggers. Encounters of these pathogens with altered innate receptors in susceptible individuals might drive pathological autoreactivity and inflammation, overcoming tolerance and causing subsequent CNS damage. In particular, functional and genetic studies reveal that Toll-like receptor (TLR) 2 and the Nod-like receptor (NLR) P3 could be involved in MS pathogenesis, whereas TLR3, the triggering receptor expressed on myeloid cells (TREM)-2 and the C-type lectin receptors (CLRs) MBL and MASP-3 would have a putative protective role. A better understanding of these interactions will provide important insights into the aetiopathogenesis of MS and could help design potential targets for novel therapies.     

Increased localization of APP‐C99 in mitochondria‐associated ER membranes causes mitochondrial dysfunction in Alzheimer disease

In the amyloidogenic pathway associated with Alzheimer disease (AD), the amyloid precursor protein (APP) is cleaved by β‐secretase to generate a 99‐aa C‐terminal fragment (C99) that is then cleaved by γ‐secretase to generate the β‐amyloid (Aβ) found in senile plaques. In previous reports, we and others have shown that γ‐secretase activity is enriched in mitochondria‐associated endoplasmic reticulum (ER) membranes (MAM) and that ER–mitochondrial connectivity and MAM function are upregulated in AD. We now show that C99, in addition to its localization in endosomes, can also be found in MAM, where it is normally processed rapidly by γ‐secretase. In cell models of AD, however, the concentration of unprocessed C99 increases in MAM regions, resulting in elevated sphingolipid turnover and an altered lipid composition of both MAM and mitochondrial membranes. In turn, this change in mitochondrial membrane composition interferes with the proper assembly and activity of mitochondrial respiratory supercomplexes, thereby likely contributing to the bioenergetic defects characteristic of AD.     

Morphological and morphometric analysis of scutella of six species and two subspecies of Triatoma (Hemiptera: Reduviidae) in Mexico

Morphological characters can be used to distinguish the vast majority of triatomine species, but the existence of high levels of phenotypic plasticity and recently diverged species can lead to erroneous determinations. To approach this problem, we analyzed the male and female morphologies of the scutella of Triatoma barberi, T. dimidiata, T. lecticularia, T. mexicana, T. recurva, T. rubida, and two sub‐species, T. protracta protracta and T. protracta nahuatlae. Scutellum samples were observed by scanning electron microscopy and subjected to morphological analysis and morphometric investigation using a canonical discriminant analysis. The results revealed differences primarily in central depression shape, posterior process, and vestiture. We observed clear dimension‐based differences in scutellum morphometry in all the taxa under study, providing sound evidence for species and subspecies differentiation. On the other hand, there is no difference between sexes in T. lecticularia, T. protracta protracta, and T. protracta nahuatlae. Our methodology can be implemented to differentiate species of the genus Triatoma.     

Characterization of a Chlorophyta microalga isolated from a microbial mat in Salar de Atacama (northern Chile) as a potential source of compounds for biotechnological applications

Microalgae are an important source of unsaturated fatty acids, phospholipids, glycolipids, and carotenes, which are useful compounds for the food and pharmaceutical industries. The Atacama Desert of northern Chile is one of the driest deserts on Earth and, as such, it is a great natural laboratory in which to study new microorganisms adapted to extreme environments. A microalgal strain, referred to here as CH03, was isolated from a microbial mat in salt flat water in Salar de Atacama. Genetic analysis of the 18S ribosomal RNA gene showed that the strain had homology with other known sequences of the species Chlorella sorokiniana. Our results revealed the adaptability of this microalga to freshwater medium under laboratory conditions, despite coming from an extremely high‐salinity environment. The fatty acid profile of CH03(A) newly isolated in Bold's basal medium differed from that of CH03(B) cultured in vitro in modified F/2 medium and from another five strains of C. sorokiniana and three strains of Chlorella vulgaris in that it had a high stearic acid content and had no polyunsaturated fatty acids. The major biochemical components observed in this strain were proteins (64.3–73.6%) and lipids (26.6–32.6%). This study suggests that the strain CH03 could be a protein source and that this oleaginous microalga is easy to grow in vitro as a biological model for future studies.     

Regulation of spiral coiling in the terrestrial gastropod Sphincterochila: An experimental test of the road-holding model

Hutchinson's ('89) road-holding model states that spiral ornaments of the snail shell (keels and low-curvature areas) dictate the growth path of the subsequent whorl, which in turn gives the signal for attachment of the next whorl. Experiments were performed with two species of the terrestrial snail Sphincterochila in order to test the role of the external keel in determining the correct coiling of successive turns. Experiments substituted a ridge made of silicone for the keel. This ridge ran either (1) abapical or (2) adapical of the original keel. In mode (1), subsequent growth continued by taking the false keel as the adapical limit of the whorl. In only very few instances of mode (2) did the whorls extend incipiently slightly adapical of the path of the original keel. Our results confirm that the keel is an important reference for the coiling strategy of the snail, although the keel itself probably does not constitute the reference, but rather the two flat ramps into which the keel divides the outer lip of the aperture. J. Morphol. 235:249–257, 1998. © 1998 Wiley-Liss, Inc.     

Development of polarity in cerebellar granule neurons

Axon formation in developing cerebellar granule neurons in situ is spatially and temporally segregated from subsequent neuronal migration and dendrite formation. To examine the role of local environmental cues on early steps in granule cell differentiation, the sequence of morphologic development and polarized distribution of membrane proteins was determined in granule cells isolated from contact with other cerebellar cell types. Granule cells cultured at low density developed their characteristic axonal and dendritic morphologies in a series of discrete temporal steps highly similar to those observed in situ, first extending a unipolar process, then long, thin bipolar axons, and finally becoming multipolar, forming short dendrites around the cell body. Axonal- and dendritic-specific cytoskeletal markers were segregated to the morphologically distinct domains. The cell surface distribution of a specific class of endogenous glycoproteins, those linked to the membrane by a glycosylphosphatidyl inositol (GPI) anchor, was also examined. The GPI-anchored protein, TAG-1, which is segregated to the parallel fiber axons in situ, was found exclusively on granule cell axons in vitro; however, two other endogenous GPI-anchored proteins were found on both the axonal and somatodendritic domains. These results demonstrate that granule cells develop polarity in a cell type-specific manner in the absence of the spatial cues of the developing cerebellar cortex. © 1997 John Wiley & Sons, Inc. J Neurobiol 32: 223–236, 1997.     

The differential NF-kB modulation by S-adenosyl-L-methionine,N-acetylcysteine and quercetin on the promotion stage of chemical hepatocarcinogenesis

S-adenosylmethionine (SAM), N-acetylcysteine (NAC) and quercetin exhibit a chemoprotective effect. Likely this effect is mediated by counteracting, oxidative stress and NF-kB activation. To test this hypothesis F344 rats were subjected to hepatocarcinogenesis with or without antioxidants. NAC decreased foci in number and area, SAM and quercetin decreased area. Lipid-peroxidation was decreased by antioxidants, but only SAM increased glutathione. SAM, in its regulation from IKK downwards, abolished the NF-kB activation. NAC decreased IKK and IkB-a phosphorylation, and Rel-A/p65 and NF-kB binding, though the last two were affected with less intensity compared to the NF-kB inhibitor. Quercetin decreased Rel-A/p65, without modifying upstream signalling. Although all antioxidants inhibited oxidative stress as shown by reduction of lipid peroxidation, not all exerted the same effect on NF-kB signalling pathway and only SAM increased GSH. The mechanisms exerted by SAM in the reduction of foci makes this compound a potential liver cancer therapeutic agent.