Showing papers by "University of Córdoba (Spain) published in 2003"

Geant4—a simulation toolkit

Abstract: G eant 4 is a toolkit for simulating the passage of particles through matter. It includes a complete range of functionality including tracking, geometry, physics models and hits. The physics processes offered cover a comprehensive range, including electromagnetic, hadronic and optical processes, a large set of long-lived particles, materials and elements, over a wide energy range starting, in some cases, from 250 eV and extending in others to the TeV energy range. It has been designed and constructed to expose the physics models utilised, to handle complex geometries, and to enable its easy adaptation for optimal use in different sets of applications. The toolkit is the result of a worldwide collaboration of physicists and software engineers. It has been created exploiting software engineering and object-oriented technology and implemented in the C++ programming language. It has been used in applications in particle physics, nuclear physics, accelerator design, space engineering and medical physics.

Engineering increased vitamin C levels in plants by overexpression of a D-galacturonic acid reductase.

Abstract: L-Ascorbic acid (vitamin C) in fruits and vegetables is an essential component of human nutrition. Surprisingly, only limited information is available about the pathway(s) leading to its biosynthesis in plants. Here, we report the isolation and characterization of GalUR, a gene from strawberry that encodes an NADPH-dependent D-galacturonate reductase. We provide evidence that the biosynthesis of L-ascorbic acid in strawberry fruit occurs through D-galacturonic acid, a principal component of cell wall pectins. Expression of GalUR correlated with changing ascorbic acid content in strawberry fruit during ripening and with variations in ascorbic acid content in fruit of different species of the genus Fragaria. Reduced pectin solubilization in cell walls of transgenic strawberry fruit with decreased expression of an endogenous pectate lyase gene resulted in lower ascorbic acid content. Overexpression of GalUR in Arabidopsis thaliana enhanced vitamin C content two- to threefold, demonstrating the feasibility of engineering increased vitamin C levels in plants using this gene.

Ultrasound: a powerful tool for leaching

Abstract: The applications of ultrasound as auxiliary energy for accelerating leaching processes are reviewed to alert analysts to its advantages. The widely used batch systems are compared with dynamic systems, the use of which is still very rare. Comparison of ultrasound-assisted leaching with other sorts of leaching, such as Soxhlet, microwave-assisted and supercritical fluid extraction, shows the potential of ultrasound for leaching in a number of areas.

Fusarium oxysporum: exploring the molecular arsenal of a vascular wilt fungus

Abstract: SUMMARY Taxonomy: Vascular wilt fungus; Ascomycete although sexual stage is yet to be found. The most closely related teleomorphic group, Gibberella, is classified within the Pyrenomycetes. Host range: Very broad at the species level. More than 120 different formae speciales have been identified based on specificity to host species belonging to a wide range of plant families. Disease symptoms: Initial symptoms of vascular wilt include vein clearing and leaf epinasty, followed by stunting, yellowing of the lower leafs, progressive wilting of leaves and stem, defoliation and finally death of the plant. In cross-sections of the stem, a brown ring is evident in the area of the vascular bundles. Some formae speciales are not primarily vascular pathogens but cause foot- and rootrot or bulbrot. Economic importance: Causes severe losses on most vegetables and flowers, several field crops such as cotton and tobacco, plantation crops such as banana, plantain, coffee and sugarcane, and a few shade trees. Control: Use of resistant varieties is the only practical measure for controlling the disease in the field. Under greenhouse conditions, soil sterilization can be performed. Alternative control methods with potential for the future include soil solarization and biological control with antagonistic bacteria or fungi. Useful websites: http://www.fgsc.net/fus.htm, http://www-genome.wi.mit.edu/annotation/fungi/fusarium/, http://www.cbs.knaw.nl/fusarium/database.html

Comparative study of the discriminating capacity of RAPD, AFLP and SSR markers and of their effectiveness in establishing genetic relationships in olive

Abstract: RAPDs, AFLPs and SSRs were compared in terms of their informativeness and efficiency in a study of genetic diversity and relationships among 32 olive cultivars cultivated in Italy and Spain. SSRs presented a higher level of polymorphism and a greater information content, as assessed by the expected heterozygosity, than AFLPs and RAPDs. The lowest values of expected heterozygosity were obtained for AFLPs, which, nevertheless were the most efficient marker system due to their capacity to reveal the highest number of bands per reaction and because of the high values achieved for a considerable number of indexes. All three techniques discriminated the genotypes very effectively, but only SSRs were able to discriminate the cultivars Frantoio and Cellina. The correlation coefficients of similarity were statistically significant for all three marker systems used but were lower for the SSR data than for RAPDs and AFLPs. For all markers a high similarity in dendrogram topologies was obtained although some differences were observed. All the dendrograms, including that obtained by the combined use of all the marker data, reflect some relationships for most of the cultivars according to their geographic diffusion. AMOVA analysis detected greater genetic differentiation among cultivars within each country than it did between the two countries.

Iron (Hydr)Oxide Crystallinity Effects on Soil Aggregation

Abstract: Differences in crystallinity may explain why total Fe (hydr)oxide content has a variable effect on aggregate stability. Therefore, surface soil samples with a range of poorly crystalline Fe (hydr)oxide contents were characterized for water-stable aggregates >0.25 mm (WSA), mean-weighted diameter (MWD), soil organic C (OC), particle-size distribution, pH, exchangeable cations, citrate/bicarbonate/dithionite (subscript d), and acid ammonium oxalate (subscript o) extractable Fe, Al, and Si. The WSA and MWD range from 23 to 95%, and 0.3 to 5.1 mm, respectively. The effects of Fe o (1.1-6.8 g kg -1 ), Fe d (3.2-19.6 g kg -1 ), OC (2.4-24.0 g kg -1 ) and clay (141-467 g kg -1 ) contents on WSA and MWD of both A and B horizons of these soils was studied using linear regression. The poorly crystalline Fe (hydr)oxide (Fe o ) and OC contents are significantly correlated with WSA in the A horizons (r 2 = 0.95, n = 6, p = 0.001, and r 2 = 0.93, n = 6, p = 0.002, respectively) and in the B horizons (r 2 = 0.73, n = 6, p = 0.029, and r 2 = 0.76, n = 6, p = 0.024, respectively). When regressed against MWD, Fe o has an r 2 of 0.89 (n = 6, p = 0.004) in the A, and 0.97 (n = 6, p = 0.000) in the B horizons. The coefficient of determination of MWD vs. OC contents is 0.98 (n = 6, p = 0.000) in the A and 0.79 (n = 6, p = 0.018) in the B horizons. Clay and Fed contents are not significantly correlated to WSA or MWD. Apparently, the Fe o component (poorly crystalline) is more effective than Fed at stabilizing soil aggregates, even though it is present in lower concentrations. The Fe o component appears more important than OC in terms of WSA and MWD for soils with relatively low soil organic matter contents.

Inorganic materials for the negative electrode of lithium-ion batteries: state-of-the-art and future prospects

Abstract: The development of advanced rechargeable batteries for efficient energy storage finds one of its keys in the lithium-ion concept. The optimization of the Li-ion technology urgently needs improvement for the active material of the negative electrode, and many recent papers in the field support this tendency. Moreover, the diversity in the chemical nature of the materials studied so far requires a systematization work to be done to envisage the main guidelines for research and future progress areas. This review starts with an analysis of the major advantages and drawbacks of the lithium metal electrode that led to the development of the Li-ion concept. The successful carbon-based materials, including numerous forms of both natural and anthropogenic origins are then mentioned. Attention is paid to the materials prepared from side products of the petroleum industry. Then, the central part of the report concerns alternative possibilities that have emerged from the use of tin, antimony and other elements that reversibly form alloys and compounds with lithium. The systematic study takes into account the chemical nature of the elements directly involved in the reversible reactions with lithium. Thus, tin and other group 14 elements, pnictides, and oxides are summarized in three consecutive sections. In the first section, particular attention will be paid to tin oxides and phosphates and the “SnSb” compounds serves as a link with the next section. Among the metal pnictides, compounds such as CoSb 3 and CrSb 2 , are described as precursors of the electroactive material in a dispersing matrix. Also phosphides, such as CoP 3 and MnP 4 , are also discussed, the latter having interesting intercalation reactions with lithium. Finally, the recently revised field of using transition metal oxides that allow a reversible Li 2 O formation on cycling is discussed. In each section, several materials evaluated in our laboratory are included.

Social and economic interest in the control of phthalic acid esters

Abstract: Excessive use of phthalate esters in industrial applications, mainly as plasticizers, has given rise to their persistent presence in consumer goods, and has raised numerous questions about their possible health effects or damage to the environment. This article presents an overview of the economic and social interest in the control of phthalate esters and in the availability of analytical methodologies for areas such as environmental and food analyses. It discusses potential contamination of the samples, usefulness of chromatographic techniques, generally after pre-treatment steps, and rare attempts to propose alternative techniques.

Immunolocalization of ghrelin and its functional receptor, the type 1a growth hormone secretagogue receptor, in the cyclic human ovary.

Abstract: Ghrelin is a novel 28-amino acid peptide identified as the endogenous ligand for the GH secretagogue receptor (GHS-R). Besides its hallmark central neuroendocrine effects in the control of GH secretion and food intake, an unexpected reproductive facet of ghrelin has recently emerged because expression of this molecule and its cognate receptor has been demonstrated in rat testis. However, whether this signaling system is present in human gonads remains to be evaluated. In this study, we have assessed the presence and cellular location of ghrelin and its functional receptor, namely the type 1a GHS-R, in the cyclic human ovary by means of immunohistochemistry using specific polyclonal antibodies. Strong ghrelin immunostaining was demonstrated in ovarian hilus interstitial cells. In contrast, ghrelin signal was not detected in ovarian follicles at any developmental stage, nor was it present in newly formed corpora lutea (CL) at very early development. However, specific ghrelin immunoreactivity was clearly observed in young and mature CL, whereas expression of the peptide disappeared in regressing luteal tissue. Concerning the cognate receptor, ovarian expression of GHS-R1a protein showed a wider pattern of tissue distribution, with detectable specific signal in oocytes as well as somatic follicular cells; luteal cells from young, mature, old, and regressing CL; and interstitial hilus cells. Of particular note, follicular GHS-R1a peptide expression paralleled follicle development with stronger immunostaining in granulosa and theca layers of healthy antral follicles. In conclusion, our results are the first to demonstrate that ghrelin and its functional type 1a receptor are expressed in the cyclic human ovary with distinct patterns of cellular location. The presence of both components (ligand and receptor) of the ghrelin signaling system within the human ovary opens up the possibility of a potential regulatory role of this novel molecule in ovarian function under physiological and pathophysiological conditions.

The pH signalling transcription factor PacC controls virulence in the plant pathogen Fusarium oxysporum

Abstract: Gene expression in fungi by ambient pH is regulated via a conserved signalling cascade whose terminal component is the zinc finger transcription factor Pacc/Rim1p We have identified a pacC orthologue in the vascular wilt pathogen Fusarium oxysporum that binds the consensus 5'-GCCAAG-3' sequence and is proteolytically processed in a similar way to PacC from Aspergillus nidulans pacC transcript levels were elevated in F oxysporum grown in alkaline conditions and almost undetectable at extreme acidic growth conditions PacC + / - loss-of-function mutants displayed an acidity-mimicking phenotype resulting in poor growth at alkaline pH, increased acid protease activity and higher transcript levels of acid-expressed polygalacturonase genes Reintroduction of a functional pacC copy into a pacC + / - mutant restored the wild-type phenotype Conversely, F oxysporum merodiploids carrying a dominant activating pacC c allele had Increased pacC transcript and protein levels and displayed an alkalinity-mimicking phenotype with reduced acid phosphatase and increased alkaline protease activities PacC + / - mutants were more virulent than the wild-type strain in root infection assays with tomato plants, whereas pacC c strains were significantly reduced in virulence We propose that F oxysporum PacC acts as a negative regulator of virulence to plants, possibly by preventing transcription of acid-expressed genes important for infection

Expression of ghrelin in the cyclic and pregnant rat ovary.

Abstract: Ghrelin, a 28-amino acid acylated peptide, has been recently identified as the endogenous ligand for the GH secretagogue receptor. Previous studies demonstrated that ghrelin, acting centrally, strongly stimulates GH release and food intake. In this study we provide novel evidence for the expression of ghrelin in the cyclic and pregnant rat ovary. Persistent expression of ghrelin gene was demonstrated in rat ovary throughout the estrous cycle, although its relative mRNA levels varied depending on the stage of the cycle, with the lowest levels in proestrus and peak expression values on diestrous d 1, i.e. during the luteal phase of the cycle. Ghrelin immunoreactivity was predominantly located in the luteal compartment of the ovary; with intense immunostaining being detected in steroidogenic cells from corpus luteum of the current cycle as well as in all generations of regressing corpora lutea. Indeed, predominant expression of ghrelin in the corpus luteum was confirmed using a pseudopregnant rat model, where maximum ghrelin mRNA levels were detected in dissected luteal tissue. To note, the cyclicity in the profile of ovarian expression of ghrelin appeared to be tissue specific, as it was not detected in the stomach, nor was it observed in terms of circulating ghrelin levels. In addition, cyclic expression of ovarian ghrelin mRNA was disrupted by blockade of the preovulatory gonadotropin surge and ovulation by means of administration of a potent GnRH antagonist. Finally, ghrelin mRNA expression was persistently detected in rat ovary throughout pregnancy, with higher levels in early pregnancy and lower expression during the later part of gestation. In conclusion, our data provide novel evidence for the expression of ghrelin in the cyclic and pregnant rat ovary. Dynamic changes in the profile of ghrelin expression were detected during the estrous cycle and throughout pregnancy, thus suggesting a precise regulation of ovarian expression of ghrelin. Overall, our present findings may represent an additional link between body weight homeostasis and female reproductive function.

Changes in protein expression profiles in bivalve molluscs (Chamaelea gallina) exposed to four model environmental pollutants.

Abstract: Proteomics has been used in the clam Chamaelea gallina as a preliminary screening of changes in protein expression caused by pollutants, potentially useful as new biomarkers. Clams were exposed in water for seven days to four model contaminants, Aroclor 1254, copper(II), tributyltin (TBT), and arsenic(III), and cytosolic fractions were initially analyzed by two-dimensional (2-D) electrophoresis in 7 cm IPG strips (pH 4-7). On average, about 1000 spots were resolved and altered expression was qualitatively detected in 9-26 spots per treatment. Aroclor 1254, Cu(II) and As(III) had a mainly upregulating effect, in contrast to TBT. Altered protein expression was confirmed in 18 cm gels (at narrow pH ranges). The 15 spots more drastically altered were excised and analyzed by mass spectrometry (MS), and four proteins were identified. Aroclor 1254 and Cu(II) upregulated putative isoforms of tropomyosin and light chain of myosin. Actin was downregulated by Aroclor and Cu(II) but upregulated by TBT and As(III), while the opposite behavior was shown by a truncated actin form, homologous to the Drosophila act87E gene product. The exclusive identification of cytoskeletal proteins could reflect their relative abundance, their prevalence in databases in molluscs, or their role as major targets of pollutant-related oxidative stress.

Intracellular Signaling Mechanisms Mediating Ghrelin-Stimulated Growth Hormone Release in Somatotropes

Abstract: Ghrelin is a newly discovered peptide that binds the receptor for GH secretagogues (GHS-R). The presence of both ghrelin and GHS-Rs in the hypothalamic-pituitary system, together with the ability of ghrelin to increase GH release, suggests a hypophysiotropic role for this peptide. To ascertain the intracellular mechanisms mediating the action of ghrelin in somatotropes, we evaluated ghrelin-induced GH release from pig pituitary cells both under basal conditions and after specific blockade of key steps of cAMP-, inositol phosphate-, and Ca 2 -dependent signaling routes. Ghrelin stimulated GH release at concentrations ranging from 10 10 to 10 6 M. Its effects were comparable with those exerted by GHRH or the GHS L-163,255. Combined treatment with ghrelin and GHRH or L-163,255 did not cause further increases in GH release, whereas somatostatin abolished the effect of ghrelin. Blockade of phospholipase C or protein kinase C inhibited ghrelininduced GH secretion, suggesting a requisite role for this route in ghrelin action. Unexpectedly, inhibition of either adenylate cyclase or protein kinase A also suppressed ghrelininduced GH release. In addition, ghrelin stimulated cAMP production and also had an additive effect with GHRH on cAMP accumulation. Ghrelin also increased free intracellular Ca 2 levels in somatotropes. Moreover, ghrelin-induced GH release was entirely dependent on extracellular Ca 2 influx through L-type voltage-sensitive channels. These results indicate that ghrelin exerts a direct stimulatory action on porcine GH release that is not additive with that of GHRH and requires the contribution of a multiple, complex set of interdependent intracellular signaling pathways. (Endocrinology 144: 5372–5380, 2003)

Class V chitin synthase determines pathogenesis in the vascular wilt fungus Fusarium oxysporum and mediates resistance to plant defence compounds.

Abstract: Chitin, a beta-1,4-linked polysaccharide of N-acetylglucosamine, is a major structural component of fungal cell walls. Fungi have multiple classes of chitin synthases that catalyse N-acetylglucosamine polymerization. Here, we demonstrate the requirement for a class V chitin synthase during host infection by the vascular wilt pathogen Fusarium oxysporum. The chsV gene was identified in an insertional mutagenesis screen for pathogenicity mutants. ChsV has a putative myosin motor and a chitin synthase domain characteristic of class V chitin synthases. The chsV insertional mutant and a gene replacement mutant of F. oxysporum display morphological abnormalities such as hyphal swellings that are indicative of alterations in cell wall structure and can be partially restored by osmotic stabilizer. The mutants are unable to infect and colonize tomato plants or to grow invasively on tomato fruit tissue. They are also hypersensitive to plant antimicrobial defence compounds such as the tomato phytoanticipin alpha-tomatine or H2O2. Reintroduction of a functional chsV copy into the mutant restored the growth phenotype of the wild-type strain. These data suggest that F. oxysporum requires a specific class V chitin synthase for pathogenesis, most probably to protect itself against plant defence mechanisms.

A first linkage map of olive (Olea europaea L.) cultivars using RAPD, AFLP, RFLP and SSR markers

Abstract: The first linkage map of the olive (Olea europaea L.) genome has been constructed using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphisms (AFLP) as dominant markers and a few restriction fragment length polymorphisms (RFLP) and simple-sequence repeats (SSR) as codominant markers. Ninety-five individuals of a cross progeny derived from two highly heterozygous olive cultivars, Leccino and Dolce Agogia, were used by applying the pseudo test-cross strategy. From 61 RAPD primers 279 markers were obtained - 158 were scored for Leccino and 121 for Dolce Agogia. Twenty-one AFLP primer combinations gave 304 useful markers - 160 heterozygous in Leccino and 144 heterozygous in Dolce Agogia. In the Leccino map 249 markers (110 RAPD, 127 AFLP, 8 RFLP and 3 SSR) were linked. This resulted in 22 major linkage groups and 17 minor groups with fewer than four markers. In the Dolce Agogia map, 236 markers (93 RAPD, 133 AFLP, 6 RFLP and 4 SSR) were linked; 27 major linkage groups and three minor groups were obtained. Codominant RFLPs and SSRs, as well as few RAPDs in heteroduplex configuration, were used to establish homologies between linkage groups of both parents. The total distance covered was 2,765 cM and 2,445 cM in the Leccino and Dolce Agogia maps, respectively. The mean map distance between adjacent markers was 13.2 cM in Leccino and 11.9 cM in Dolce Agogia, respectively. Both AFLP and RAPD markers were homogeneously distributed in all of the linkage groups reported. The stearoyl-ACP desaturase gene was mapped on linkage group 4 of cv. Leccino.

Lead and lead toxicity in domestic and free living birds.

Abstract: At present, domestic and wild fauna are being exposed to aspects and factors which are foreign to the habitat in which they live. One that stands out is the enormous amount and variety of chemical compounds which, in many cases, are highly complex and which are constantly being released into the atmosphere, mainly from agricultural and industrial activity. All these substances affect some species more than others, whether they be plants or animals, from the most insignificant micro-organism to the most evolved species, among them birds. Finally, another cause of mortality in many birds is plumbism, namely death caused by the ingestion of lead. Lead has been one of the main causes of poisoning in man since ancient times due to its use in many activities although it is only recently that this toxicity has been recognized. Moreover, the use of lead pellets for shooting has resulted in the release into the environment of millions of these over many years, with serious repercussions for many bird species populations, which have ingested them either directly or indirectly. Added to this use of lead in cynegetic activities is the fate of the lead weights (sinkers or ballast) used by rod fishers, which sink to the bottom or accumulate on the banks of rivers, lakes, lagoons or reservoirs. The problem arises when these pellets or weights are ingested by birds, mainly Anatidae, which mistake them for the small stones or grit they use to triturate food in their gizzards. Small particles of lead enter the digestive tract, start dissolving in the form of lead salts, are incorporated into the bloodstream and the rest of the body, accumulate in organs like the liver or kidneys, and cause physiological or behavioural changes. When certain concentrations of lead are reached, the birds then die. If lead-poisoned birds are consumed by carrions or predators, the latter also ingest the lead so that they may also be affected or die from plumbism since, being a heavy metal, its degradation and/or elimination is very difficult. There is, therefore, no doubt that millions of birds die annually worldwide from lead poisoning (in the U.S.A., around 3,000,000), this problem being most acute in marshland. The solutions could include the introduction of legislation regulating or banning shooting, in the use of non-toxic ammunition in marshes and protected areas, the substitution of lead pellets for other non-toxic ones, such as steel, bismuth, tungsten or other suitable metals, and to go on studying other possible alternatives to end such a dramatic situation for birds all over the world.

Influence of the preparation method on the structural and surface properties of various magnesium oxides and their catalytic activity in the Meerwein–Ponndorf–Verley reaction

Abstract: The Meerwein–Ponndorf–Verley (MPV) reaction of cyclohexanone with isopropyl alcohol, which yields cyclohexanol, was studied by using basic catalysts consisting of magnesium oxide. The oxides were prepared using various synthetic procedures including calcination of commercially available magnesium hydroxide and carbonate, calcination of magnesium hydroxides obtained from magnesium nitrate and sulphate, the sol–gel technique and precipitation by decomposition of urea. The solids thus obtained were characterized structurally by X-ray diffraction spectroscopy and their surface properties determined using gas adsorption measurements (of nitrogen for textural properties and carbon dioxide for basic properties). Based on the results, the most active catalyst was the solid prepared by rehydration and subsequent calcination of a magnesium oxide that was previously obtained from commercially available magnesium hydroxide.

Anandamide Inhibits Nuclear Factor-κB Activation through a Cannabinoid Receptor-Independent Pathway

Abstract: Anandamide (arachidonoylethanolamine, AEA), an endogenous agonist for both the cannabinoid CB(1) receptor and the vanilloid VR1 receptor, elicits neurobehavioral, anti-inflammatory, immunomodulatory, and proapoptotic effects. Because of the central role of nuclear factor-kappaB (NF-kappaB) in the inflammatory process and the immune response, we postulated that AEA might owe some of its effects to the suppression of NF-kappaB. This study shows that AEA inhibits tumor necrosis factor-alpha (TNFalpha)-induced NF-kappaB activation by direct inhibition of the IkappaB kinase (IKK)beta and, to a lesser extent, the IKKalpha subunits of kappaB inhibitor (IkappaB) kinase complex, and that IKKs inhibition by AEA correlates with inhibition of IkappaBalpha degradation, NF-kappaB binding to DNA, and NF-kappaB-dependent transcription in TNFalpha-stimulated cells. AEA also prevents NF-kappaB-dependent reporter gene expression induced by mitogen-activated protein kinase kinase kinase and NF-kappaB-inducing kinase. The NF-kappaB inhibitory activity of AEA was independent of CB(1) and CB(2) activation in TNFalpha-stimulated 5.1 and A549 cell lines, which do not express vanilloid receptor 1, and was not mediated by hydrolytic products formed through the activity of the enzyme fatty acid amide hydrolase. Chemical modification markedly affected AEA inhibitory activity on NF-kappaB, suggesting rather narrow structure-activity relationships and the specific interaction with a molecular target. Substitution of the alkyl moiety with less saturated fatty acids generally reduced or abolished activity. However, replacement of the ethanolamine "head" with a vanillyl group led to potent inhibition of TNFalpha-induced NF-kappaB-dependent transcription. These findings provide new mechanistic insights into the anti-inflammatory and proapoptotic activities of AEA, and should foster the synthesis of improved analogs amenable to pharmaceutical development as anti-inflammatory agents.

Mixed infections and insect–pathogen interactions

Abstract: Most studies of insect–pathogen interactions consider the direct interaction between one disease agent and one species of host. However, given that hosts are subject to challenge from many pathogen ⁄ parasite species, mixed infections are probably common. In this study, using the desert locust and two species of fungal entomopathogen, we show how mixed infection with a largely avirulent pathogen can alter the virulence and reproduction of a second, highly virulent pathogen. We find that two strains of the avirulent pathogen vary in their interaction with the virulent pathogen, depending on the order of infection and environmental conditions. We propose that avirulent pathogens, which have largely been overlooked to date, could play a significant role in host– pathogen dynamics, with implications for biological control and evolution of virulence.

Cloning and characterization of two ripening‐related strawberry (Fragaria × ananassa cv. Chandler) pectate lyase genes

Abstract: Two genomic clones corresponding to putative pectate lyase genes (plA and plB) were isolated and characterized in strawberry (Fragaria x ananassa cv. Chandler). The corresponding ORFs for the plA and plB genes revealed deduced proteins of 451 and 439 amino acids, respectively, that differ from that of the previously isolated strawberry plC gene. Southern blot analysis has shown that while the plB gene is a single copy gene, the plA gene is probably encoded by a small multigene family. By using specific probes corresponding to the untranslated 3' terminal region of the pl genes, and QRT-PCR methodology, the spatio-temporal expression pattern of both strawberry pl genes have been compared with that of the plC gene. The three transcripts were specifically expressed only in fruit and mainly during the ripening stages. Moreover, the expression of the plA and plB genes was induced in green de-achened fruit, but this increase was reduced by the external application of auxins as was the expression of plC. The expression of both pl genes was also strongly reduced in harvested fruit kept in controlled atmosphere (CA) containing high CO(2) levels. Immunolocalization studies using antibodies raised against the strawberry PL proteins placed the proteins in the cell wall of parenchymatic cells of the fruit receptacle. The role of pl genes in cell-wall disassembly and fruit ripening softening is discussed.

Hydromaghemite, an intermediate in the hydrothermal transformation of 2-line ferrihydrite into hematite

Abstract: The hydrothermal transformation of 2-line ferrihydrite into hematite proceeds slowly if a suffi- cient quantity of some strongly adsorbing ligand, such as phosphate or citrate, is sorbed on the starting product. In this work, we studied such transformation at temperatures ranging from 125 to 200 ∞C, a molar P/Fe ratio of 0-6%, and a molar citrate/Fe ratio of 3%. The products were character- ized by X-ray diffraction (XRD), infrared spectroscopy, Mossbauer spectroscopy (MS) at various temperatures and in an applied field of 60 kOe, magnetic and thermal analysis, and transmission electron microscopy (TEM). At 150 ∞C, pure 2-line ferrihydrite transformed rapidly into hematite. The products of transformation of 2-line ferrihydrite with P/Fe = 2.75% or citrate/Fe = 3% had a magnetic susceptibility of >240 ∞ 10 -6 m 3 /kg and were, according to XRD and MS data, mixtures of hematite with structural P, 6-line ferrihydrite, and a magnetic phase. This phase exhibited most of the characteristic reflections and MS features of maghemite, and occurred as 7-30 nm subrounded particles with lattice fringes corresponding to the maghemite (310) and (220) planes. It was desig- nated "hydromaghemite" because it lost >3% water between 110 and ~350 ∞C. At 150 ∞C, complete transformation into hematite occurred in <120 days.

A Morphologic and Immunohistochemical Study of the Bronchus-associated Lymphoid Tissue of Pigs Naturally Infected with Mycoplasma hyopneumoniae

Abstract: Porcine enzootic pneumonia (PEN), caused by Mycoplasma hyopneumoniae (Mh), has been described in pigs in all geographic areas. The disease is characterized by high morbidity and low mortality rates in intensive swine production systems. A morphologic and immunohistochemical study was done to determine the cellular populations present in lung parenchyma of infected pigs, with special attention to the bronchus-associated lymphoid tissue (BALT). Polyclonal and monoclonal antibodies were used for the detection of antigens of Mh, T lymphocytes (CD3+, CD4+, and CD8+), IgG+ or IgA+ lymphocytes, and cells containing lysozyme, S-100 protein, major histocompatibility complex class II antigen or myeloid-histiocyte antigen. Findings in lung tissues associated with Mh infection were catarrhal bronchointerstitial pneumonia, with infiltration of inflammatory cells in the lamina propria of bronchi and bronchioles and alveolar septa. Hyperplasia of mononuclear cells in the BALT areas was the most significant histologic change. The BALT showed a high morphologic and cellular organization. Macrophages and B lymphocytes were the main cellular components of germinal centers. T lymphocytes were primarily located in perifollicular areas of the BALT, lamina propria and within the airway epithelium, and plasma cells containing IgG or IgA at the periphery of the BALT, in the lamina propria of bronchi and bronchioles, in alveolar septa, and around bronchial submucosal glands. The hyperplastic BALT in PEN cases consisted of macrophages, dendritic cells, T and B lymphocytes, and IgG+ and IgA+ plasma cells. CD4+ cells predominated over CD8+ cells. Local humoral immunity appears to play an important role in the infection.

Characterization of potassium transport in wild-type and isogenic yeast strains carrying all combinations of trk1, trk2 and tok1 null mutations.

Abstract: Saccharomyces cerevisiae cells express three defined potassium-specific transport systems en-coded by TRK1, TRK2 and TOK1. To gain a more complete understanding of the physiological function of these transport proteins, we have constructed a set of isogenic yeast strains carrying all combinations of trk1delta, trk2delta and tok1delta null mutations. The in vivo K+ transport characteristics of each strain have been documented using growth-based assays, and the in vitro biochemical and electrophysiological properties associated with K+ transport have been determined. As has been reported previously, Trk1p and Trk2p facilitate high-affinity potassium uptake and appear to be functionally redundant under a wide range of environmental conditions. In the absence of TRK1 and TRK2, strains lack the ability specifically to take up K+, and trk1deltatrk2delta double mutant cells depend upon poorly understood non-specific cation uptake mechanisms for growth. Under conditions that impair the activity of the non-specific uptake system, termed NSC1, we have found that the presence of functional Tok1p renders cells sensitive to Cs+. Based on this finding, we have established a growth-based assay that monitors the in vivo activity of Tok1p.