Identification of yeast genes that confer resistance to chitosan oligosaccharide (COS) using chemogenomics

Background: Chitosan oligosaccharide (COS), a deacetylated derivative of chitin, is an abundant, and renewable natural polymer. COS has higher antimicrobial properties than chitosan and is presumed to act by disrupting/permeabilizing the cell membranes of bacteria, yeast and fungi. COS is relatively non-toxic to mammals. By identifying the molecular and genetic targets of COS, we hope to gain a better understanding of the antifungal mode of action of COS. Results: Three different chemogenomic fitness assays, haploinsufficiency (HIP), homozygous deletion (HOP), and multicopy suppression (MSP) profiling were combined with a transcriptomic analysis to gain insight in to the mode of action and mechanisms of resistance to chitosan oligosaccharides. The fitness assays identified 39 yeast deletion strains sensitive to COS and 21 suppressors of COS sensitivity. The genes identified are involved in processes such as RNA biology (transcription, translation and regulatory mechanisms), membrane functions (e.g. signalling, transport and targeting), membrane structural components, cell division, and proteasome processes. The transcriptomes of control wild type and 5 suppressor strains overexpressing ARL1, BCK2, ERG24, MSG5, or RBA50, were analyzed in the presence and absence of COS. Some of the up-regulated transcripts in the suppressor overexpressing strains exposed to COS included genes involved in transcription, cell cycle, stress response and the Ras signal transduction pathway. Down-regulated transcripts included those encoding protein folding components and respiratory chain proteins. The COS-induced transcriptional response is distinct from previously described environmental stress responses (i.e. thermal, salt, osmotic and oxidative stress) and pre-treatment with these well characterized environmental stressors provided little or any resistance to COS. Conclusions: Overexpression of the ARL1 gene, a member of the Ras superfamily that regulates membrane trafficking, provides protection against COS-induced cell membrane permeability and damage. We found that the ARL1 COS-resistant over-expression strain was as sensitive to Amphotericin B, Fluconazole and Terbinafine as the wild type cells and that when COS and Fluconazole are used in combination they act in a synergistic fashion. The gene targets of COS identified in this study indicate that COS’s mechanism of action is different from other commonly studied fungicides that target membranes, suggesting that COS may be an effective fungicide for drug-resistant fungal pathogens.

Elysia timida (Risso, 1818) three decades of research

During the last 30 years, studies on Elysia timida (Risso, 1818) have addressed various aspects related to food sources, photosynthetic efficiency of kleptoplasts, population genetics, chemical ecology and reproductive biology, both in the Mediterranean Sea and in the Mar Menor coastal lagoon. E. timida shows a strong specific interaction with Acetabularia acetabulum, retaining functional chloroplasts for at least 45 days and obtaining extra energy in periods when food resources are scarce. It shows control of parapodia, avoiding pigment photodestruction under oversaturated light conditions. The chemical ecological relationships established between E. timida and its potential predator fish, Thalassoma pavo, have also been evaluated, and it has been found that that the extracts of the mollusc contain repellent and unpalatable polypropionate compounds. Population genetics has demonstrated the genetic divergence between populations showing high and significant values of FST and genetic distances, and at least six privative alleles that are not shared with Mediterranean populations have been detected in lagoon populations. This sacoglossan is a poecilogonic species, and its lagoon populations show a greater reproductive output than Mediterranean populations; they produce a greater number of egg masses and embyros per individual, and the capsules have a wider diameter.

New Mediterranean Biodiversity Records (June 2012)

The present work reports on the extended distribution of nineteen species in the Mediterranean. These are: Upeneus pori (Fish:Turkey), Bursatella leachii (Mollusca, Opisthobranchia: eastern coast of Spain), Sparisoma cretense (Fish: Ionian coast of Greece), Pseudobryopsis myura (Chlorophyta:Turkey), Aplysia dactylomela (Mollusca, Opisthobranchia: Karpathos island, and Kyklades Archipelago, Greece), Asparagopsis armata and Botryocladia madagascariensis (Rhodophyta: South Peloponnesos, Greece), Oxynotus centrina (Fish: Greece), Caulerpa racemosa var. cylindracea (Chlorophyta ), Stypopodium schimperi (Phaeophyta ) Siganus luridus and Stephanolepis diaspros (Fish) Percnon gibbesi (Decapoda, Brachyura) (Kyklades Archipelago, Greece), Cerithium scabridum (Mollusca, Prosobranchia: Anavissos: Greece) and Cerithium renovatum (Mollusca, Prosobranchia: N. Κriti), Cassiopea andromeda (Scyphomedusa: Rhodos Island, Greece), Abra tenuis (Mollusca Bivalvia: Vouliagmeni Lake, Greece) Lagocephalus lagocephalus (Fish: Calabrian coast, Italy) and Plocamopherus ocellatus (Mollusca, Opisthobranchia: İskenderun Bay, Turkey).

Checking the concurrence among macrobenthic organism distribution patterns at different taxonomic scales in relation to environmental factors

The interaction of both natural conditions and anthropogenic environmental impacts can lead to different soft-bottom macrobenthic distribution patterns. Soft-bottom macrobenthic community was analysed at different taxonomic scales in order to evaluate whether diverse subset of organisms respond to the variability of the environmental pressures (natural and human induced) showing or not similar distribution patterns. Therefore, this long-term survey had been focused on a heterogeneous area, where both anthropogenic and natural stress may affect the community. Three perpendicular transects to the coast were established and stations at 4, 10 and 15 m depths were sampled at each transect twice a year (summer- winter) from 2004 to 2009. Non-parametric multivariate techniques were used to analyse soft-bottom macrobenthic community distribution and its relation to the environmental factors. Similar distribution patterns between investigated taxonomic levels were detected and they were mainly related to depth.

Colour morphotypes of Elysia timida (Sacoglossa, Gastropoda) are determined by light acclimation in food algae

Elysia timida (Risso, 1818) colonizing the shallow waters of the Mar Menor Lagoon (Spain) exhibit a brown and a green morph. It was hypothesised that these morphs were the result of feeding preferentially on brown and green algae, respectively. E. timida and its potential food sources, Acetabularia acetabulum (Chlorophyta) and Halopteris filicina (Heterokontophyta) were collected by snorkelling during April 2010. Photosynthetic pigments were analysed by HPLC, photo-physiological parameters were estimated by PAM fluorometry and body colour was characterized by spectral reflectance. Digital photography was used to count the number and area of red spots (small red dots on the slug’s surface) on the parapodia of the 2 morphs. In the laboratory, green E. timida was fed with A. acetabulum cultured under 2 light treatments (high light, 600 µmol E m−2 s−1 and low light, 40 µmol E m−2 s−1), and digital photography was used to monitor colour alterations in E. timida. Spectral reflectance confirmed the colour differences, but both morphs showed a pigment composition similar to the green alga A. acetabulum and showed none of the pigments present in the brown alga H. filicina, neither immediately after collection of the slugs in situ, nor after the feeding experiment. A. acetabulum grown under high light intensity changed from green to brown colour and E. timida changed to brown colour when fed with high-light acclimated A. acetabulum. Thus, E. timida colour differences could not be attributed to feeding on different algae groups but was likely the result of feeding on A. acetabulum growing under different light intensities.

Fish diversity and traditional fishing methods in semiarid irrigated system in the Bajo Vinalopó region, Southeastern Spain

The aim of this paper is to obtain a comprehensive picture of the importance of the fish fauna in the wetlands and coastal lagoons of the Bajo Vinalopó region (Alicante, South eastern Spain) and the ecological, historical and cultural value of the traditional knowledge on fishing maintained by local people. We have compiled data from different information sources: (1) the revision of historical archives, (2) personal interviews and collaborative research, and (3) ecological and water quality data. The results show that the area sustains important fish fauna diversity (17 species) and that local people have inherited a considerable traditional knowledge on fishing methods (11 modalities), on the maintenance and sound management of the water system and on the ecology and behaviour of fish. We conclude that a comprehensive consideration of all these ecological, historical and socio-cultural aspects related to fish and fishing shows clearly the value of this ecological and cultural heritage and provides a necessary base for a sustainable management of the area.

Using the damage from 2010 Haiti earthquake for calibrating vulnerability models of typical structures in Port-au-Prince (Haiti)

After the 2010 Haiti earthquake, that hits the city of Port-au-Prince, capital city of Haiti, a multidisciplinary working group of specialists (seismologist, geologists, engineers and architects) from different Spanish Universities and also from Haiti, joined effort under the SISMO-HAITI project (financed by the Universidad Politecnica de Madrid), with an objective: Evaluation of seismic hazard and risk in Haiti and its application to the seismic design, urban planning, emergency and resource management. In this paper, as a first step for a structural damage estimation of future earthquakes in the country, a calibration of damage functions has been carried out by means of a two-stage procedure. After compiling a database with observed damage in the city after the earthquake, the exposure model (building stock) has been classified and through an iteratively two-step calibration process, a specific set of damage functions for the country has been proposed. Additionally, Next Generation Attenuation Models (NGA) and Vs30 models have been analysed to choose the most appropriate for the seismic risk estimation in the city. Finally in a next paper, these functions will be used to estimate a seismic risk scenario for a future earthquake.

Evaluation of the pathogenicity of multiple isolates of Beauveria bassiana (Hypocreales: Clavicipitaceae) on Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae) for the assessment of a solid formulation under simulated field conditions

A solid state formulation of Beauveria bassiana (Balsamo) Vuillemin has been developed for biological control of the Red Palm Weevil (RPW), Rhynchophorus ferrugineus (Olivier, 1790). Two kinds of bioassays (dry conidia and dipping) using 10 isolates from several coleopterans in Mediterranean environments, identified 2 RPW derived isolates (193 and 203) as most pathogenic to RPW larvae and adults (zero survival within first 4–5 d for dry conidia, and 14 and 23 d for dipping bioassays). Isolate 203 (5.1 × 108 ± 1.9 × 108 conidia g-1) was formulated with fragmented date seed into solid granules and tested in palms infested with RPW under semi-field conditions in Feb, Apr/May and Jun of both 2007 and 2008. Beauveria bassiana significantly reduced RPW adult survival with respect to controls in May 2007 and in the Apr/Jun 2008 experiments. Total RPW adult mortality was achieved within 30 days for all B. bassiana treatments, and was associated with increasing numbers of insects with signs of mycosis in 2008 experiments. Beauveria bassiana formulation reduced RPW multiplication in artificially infested palms compared to controls, and a positive correlation between numbers of larvae and time post-infestation was recorded. The suppression of RPW adult populations by B. bassiana persisted for at least 3 months under semi-field conditions. The Beauveria bassiana solid formulation, which induces great adult mortality and persistence in the field, could be applied as a preventive as well as a curative treatment for the integrated management of RPW.

Sequencing and functional analysis of the genome of a nematode egg-parasitic fungus, Pochonia chlamydosporia

Pochonia chlamydosporia is a worldwide-distributed soil fungus with a great capacity to infect and destroy the eggs and kill females of plant-parasitic nematodes. Additionally, it has the ability to colonize endophytically roots of economically-important crop plants, thereby promoting their growth and eliciting plant defenses. This multitrophic behavior makes P. chlamydosporia a potentially useful tool for sustainable agriculture approaches. We sequenced and assembled ∼41 Mb of P. chlamydosporia genomic DNA and predicted 12,122 gene models, of which many were homologous to genes of fungal pathogens of invertebrates and fungal plant pathogens. Predicted genes (65%) were functionally annotated according to Gene Ontology, and 16% of them found to share homology with genes in the Pathogen Host Interactions (PHI) database. The genome of this fungus is highly enriched in genes encoding hydrolytic enzymes, such as proteases, glycoside hydrolases and carbohydrate esterases. We used RNA-Seq technology in order to identify the genes expressed during endophytic behavior of P. chlamydosporia when colonizing barley roots. Functional annotation of these genes showed that hydrolytic enzymes and transporters are expressed during endophytism. This structural and functional analysis of the P. chlamydosporia genome provides a starting point for understanding the molecular mechanisms involved in the multitrophic lifestyle of this fungus. The genomic information provided here should also prove useful for enhancing the capabilities of this fungus as a biocontrol agent of plant-parasitic nematodes and as a plant growth-promoting organism.

Inventario de plantas con interés etnobotánico de Carrizales (Elche), SE de la Península Ibérica

Se ha realizado un inventario de flora útil en la Partida de Carrizales en el término municipal de Elche (Alicante). A partir de entrevistas de tipo semiestructurado se han encontrado un total de 246 especies, de las cuales aparece una pequeña descripción botánica, ecológica y aplicación etnobotánica. Así, se pretende hacer una aproximación al conocimiento etnobotánico existente en una zona de regadío colonizada a partir del s. XVIII e intensamente antropizada. En nuestro trabajo hemos prestado atención no sólo a los cultivos que actualmente encontramos, sino también los que han estado presentes en el pasado, ya que conociendo sus características se pueden tratar de recuperar y crear productos con sello propio y dinamizar la economía local.

El Plan Bonet, 1961-64. La condición del paisaje en la ordenación del desarrollo turístico de La Manga del Mar Menor

En el contexto rural de Murcia, la recepción de la modernidad fue un fenómeno episódico y tardío. Hacia 1960 se produjo, sin embargo, un breve e ilusionante periodo cuando algunos maestros de la arquitectura española llegaron para trabajar en el desarrollo turístico de un litoral virgen y lleno de oportunidades. Entre ellos, Antoni Bonet recibió el encargo de urbanizar La Manga, una estrecha franja de tierra de 24km que separa el Mediterráneo del Mar Menor. El Plan retomaba los planteamientos urbanos de Le Corbusier para reelaborar algunas ideas preparatorias: autonomía y jerarquía de los núcleos habitados, eje vertebrador, centro cívico al borde del mar, tratamiento del frente marítimo con islas artificiales. Minimizando su impacto en el ecosistema dunar con una carretera elevada puntualmente para favorecer el aporte de arena, el Plan concentraba la edificación alrededor de torres de 16 plantas, generando unidades compactas de viviendas y equipamientos repetidas cada 2km. Inspirándose en la lectura del territorio de Le Corbusier en Buenos Aires (1929), Bonet respondía a la horizontalidad del plano del mar por contraste topológico, subrayando la presencia vertical de las torres como hitos paisajísticos que, desde la orilla opuesta, marcaban la posición de cada núcleo.

Evolution of Posidonia oceanica seagrass meadows and its implications for management

Results of the monitoring network of the Posidonia oceanica meadows in the Valencia region in Spain are analysed. For spatial comparison the whole data set has been analysed, however, for temporal trends we only selected stations that have been monitored at least 6 years in the period of 2002–2011 (26 stations in 13 localities). At the south of the studied area, meadows are larger, and they have higher density and covering than that in the Valencia Gulf, excluding Oropesa meadow. Monitoring of P. oceanica meadows in the Valencia region in Spain indicates that most of them are stationary or they are increasing their density and covering while no decline was observed in the studied meadows. These results indicate that there is not a general decline of P. oceanica meadows and that the decline of P. oceanica, when it has been observed in other studies, is produced by local causes that may be managed at the local level. This study also reflects the importance of long series of direct data to analyse trends in the population dynamics for slow-growing species.

Análisis y modelización de la actividad pesquera. La flota española en las aguas internacionales de la plataforma patagónica

Esta tesis está enfocada en el programa de observadores científicos en aguas del Atlántico Sudoccidental que el Instituto Español de Oceanografía (IEO) mantiene desde el año 1989 hasta la actualidad. Dicha base de datos ya ha sido empleada previamente por otros autores para estudiar los recursos pesqueros presentes, tanto en las aguas internacionales de la plataforma patagónica, más allá de las 200 millas náuticas de la costa argentina, como en la zona de pesca actualmente incluida dentro de la jurisdicción de las islas Falkland/Malvinas. Aquí nos centramos en el estudio de la actividad pesquera española que faena en la zona de aguas internacionales conocida como “División 46” y de los recursos a los que se dirige. Varios factores justifican la necesidad de esta tesis: (1) La necesidad de desarrollar una descripción detallada de la base de datos, no realizada hasta el momento. (2) Valorar la representatividad espacial del programa de observadores. (3) Establecer una serie de análisis espacio-temporales que describan la actividad pesquera de la flota española en estas aguas, (4) Desarrollar un completo y detallado análisis espacio-temporal de la distribución y abundancia de los recursos pesqueros a los que se dirige la flota. Por otra parte, las limitaciones propias de un programa de observadores científicos en buques pesqueros nos llevan, por necesidad, a explorar soluciones metodológicas que permitan un análisis fiable del esfuerzo pesquero y de la abundancia de los recursos. A este respecto, la disponibilidad de nueva información relacionada con las características físicas del área de estudio obtenida a raíz de las campañas oceanográficas realizadas en las aguas internacionales de la plataforma patagónica por el IEO y la Secretaría General del Mar (SGM) entre los años 2007 y 2011 y la disponibilidad de datos procedentes del Sistema de Localización de Buques de la SGM entre el periodo 2001 y 2007, nos ha permitido avanzar en el conocimiento de la actividad pesquera española en esta región. Como resultados principales, hemos definido algunas características fundamentales que determinan el marco de estudio de esta base de datos, hemos explorado diversas aproximaciones metodológicas que permitan un análisis de la representatividad de un programa de observadores en aguas internacionales, hemos desarrollado una descripción espacio-temporal de la estrategia de pesca en esta aguas y, a raíz del conocimiento adquirido, hemos realizado un análisis de los factores que afectan a la distribución de M. hubbsi y al esfuerzo de pesca invertido en su captura.

Single-cell derived clones from human adipose stem cells present different immunomodulatory properties

Human adipose mesenchymal stem cells are a heterogeneous population, where cell cultures derived from single cell-expanded clones present varying degrees of differential plasticity. This work focuses on the immunomodulatory/anti-inflammatory properties of these cells. To this end, 5 single cell clones were isolated (generally called 1.X and 3.X) from 2 volunteers. Regarding the expression level of the lineage-characteristic surface antigens, clones 1.10 and 1.22 expressed the lowest amounts, while clones 3.10 and 3.5 expressed more CD105 than the rest and clone 1.7 expressed higher amounts of CD73 and CD44. Regarding cytokine secretion, all clones were capable of spontaneously releasing high levels of IL-6 and low to moderate levels of IL-8. These differences can be explained in part by the distinct methylation profile exhibited by the clones. Furthermore and after lipopolysaccharide stimulation, clone 3.X produced the highest amounts of pro-inflammatory cytokines such as IL-1β, while clones 1.10 and 1.22 highly expressed IL-4 and IL-5. In co-culture experiments, clones 1.X are altogether more potent inhibitors than clones 3.X for proliferation of total, CD3+T, CD4+T and CD8+T lymphocytes and NK cells. The results of this work indicates that adipose stem cell population is heterogeneous in cytokine production profile, and that isolation, characterization and selection of the appropriate cell clone is a more exact method for the possible treatment of different patients or pathologies.