Model experiments for selection on resistance to infection in mice and pigs

International audienc

Krefelderski test - novi eksperimentalni postupak za proučavanje toksičnosti kemikalija

During the last years, skin compatibility of textiles has gained importance for consumers. Therefore, the development of suitable test methods for the evaluation of skin compatibility is a vital challenge. The fundamental requirements for these test methods are low experimental complexity combined with minor costs. The Cilitatetest has been developed for the cosmetic and pharmaceutics industry. Due to the optical measurement of the number of living microorganisms, this method is not suitable for colored or heterogeneous samples. However, using a calorimetric detection, these problems are resolved. This method allows very sensitive detection of the metabolism of the microorganism. The presence of any toxic substance results in a reduction of the heat of metabolism.Kompatibilnost tekstila prema koži postajala je tijekom posljednjih godina sve važnija potrošačima. Zato je razvoj prikladnih postupaka ispitivanja za prosudbu kompatibilnosti s kožom primjeran izazov. Osnovni zahtjev postavljen za ove postupke ispitivanja jest eksperimentalna jednostavnost povezana s nižim troškovima. Ispitivanje trepetljikašima razvijeno je za kozmetičku i farmaceutsku industriju. Zbog optičkog mjerenja broja živih mikroorganizama taj postupak nije prikladan za obojene ili heterogene uzorke. Međutim, ti problemi su riješeni primjenom kalorimetijskog otkrivanja. Taj postupak otkrivanja omogućuje vrlo osjetljivo prepoznavanje metabolizma mikroorganizama. Postojanje bilo koje otrovne tvari znači smanjenje topline metabolizm

Short term effects of irradiance on the growth of Pterocladiella capillacea (Gelidiales, Rhodophyta)

Pterocladiella capillacea has been economically exploited for agar extraction in the Azores for many years. Harvesting dropped to a full stop in the early 1990s due to a population collapse, but restarted in 2013. Since then it has been intensively harvested and overexploitation must be prevented, with both sustainable harvesting and effective cultivation practices. This study represents the first attempt to determine optimal conditions for P. capillacea production in the Azores, and evaluates its vegetative growth in two experiments using von Stosch’s medium designed to test entire thallus and tips portions response to different irradiances (30, 70 and 150 μmol photons m¯² s¯¹). The best relative growth rate (RGR) was recorded at 150 μmol photons m¯² s¯¹ for the entire thalli and tips after two-weeks and three-weeks, respectively, indicating that an acclimation period is necessary to assure the growth of this alga under experimental conditions. Higher RGR was obtained at higher irradiance (3.98 ± 2.10% fm day¯¹), but overall, growth rates were low or negative. Epiphytes were a serious problem towards the end of the entire thallus experiments, where Feldmannia irregularis proliferate at all irradiances. Future cultivation approaches complemented with other relevant environmental factors (e.g. pH, photoperiod, salinity), are recommended.FCT – Fundação para a Ciência e Tecnologia projects UID/BIA/00329/2013, 2015 - 2018 and UID/BIA/00329/2019, CIRN (Centro de Investigação de Recursos Naturais, University of the Azores), and CIIMAR (Interdisciplinary Centre of Marine and Environmental Research, Porto, Portugal). RFP was supported by a doctoral grant M3.1.2/F/024/2011, Fundo Regional para a Ciência e Tecnologia.info:eu-repo/semantics/publishedVersio

Dissipationsintegralverfahren für turbulente Grenzschichten

Mit dieser Arbeit liegt eine ausführliche Studie zu den Integralverfahren der Grenzschichttheorie vor. Der Schwerpunkt liegt dabei auf den Dissipationsintegralverfahren. Im Vergleich zu anderen Grenzschichtverfahren bestehen die generellen Vorteile der Integralverfahren in ihrer Robustheit, ihrer hohen Praktikabilität sowie den im Sinne der aufzuwendenden Computerkapazität geringen Kosten. Ein spezieller Vorteil der Dissipationsintegralverfahren ist es zudem, das komplette Schubspannungsprofil der Grenzschicht zu berücksichtigen. Über eine entsprechende Ableitung der Dissipationsintegrale kann die Vorgeschichte der Grenzschicht erfassen werden. Ausgangspunkt der Arbeit ist eine Literaturanalyse, welche aufzeigt, dass Integralverfahren weit verbreitet sind und für vielfältige Typen von Grenzschichten Anwendung finden. Ausgehend von den Navier-Stokes-Gleichungen wird über die Grenzschichtgleichungen die allgemeine Form der Dissipationsintegralgleichungen hergeleitet. Auf der Basis dieser Gleichungen werden Berechnungsalgorithmen für zwei- und dreidimensionale Grenzschichten entworfen und ausführlich diskutiert. Die zur Komplettierung der Berechnungsalgorithmen benötigten Parameterzusammenhänge werden aus expliziten Geschwindigkeitsprofilen hergeleitet. Hierzu werden beruhend auf dem Zweischichtenmodell turbulenter Grenzschichten Geschwindigkeitsprofile turbulenter Grenzschichten diskutiert. Drei Kombinationen von Haupt- und Querströmungsprofil werden für den dreidimensionalen Fall ausgewählt und algorithmisch umgesetzt. Die Algorithmen für zweidimensionale Grenzschichten beruhen auf einem Geschwindigkeitsprofil. Dem Aufbau der Algorithmen schließen sich ausführliche Testrechnungen sowie eine Bewertung der Verfahren an. Es wird festgestellt, dass Dissipationsintegralverfahren für zwei- und dreidimensionale Grenzschichten mit gutem Erfolg angewandt werden können. Vergleiche der Rechenergebnisse für zweidimensionale Grenzschichten zeigen die zumindest Gleichwertigkeit mit Zweigleichungsmodellen an.Calculation and prediction of turbulent boundary layers are among the most challenging tasks of present fluid mechanics. A strong demand exists for robust, easy-to-handle and in terms of computing effort cheap algorithms which can be used for technical applications. From an engineering point of view zonal methods and RANS are the most useful tools for solution of fluid mechanical problems. It is known that zonal methods which use integral approaches for the description of the boundary layer can be used successfully in manifold forms. One way to improve zonal methods further is the Basically three different types of integral algorithms - entrainment, momentum of momentum and dissipation integral method - can be derived from the three-dimensional boundary layer equations. If one compares the usual entrainment integral method with the dissipation integral method it turns out that the latter has the following physical advantages. While the entrainment method considers information about the shear stress distribution only at the outer edge of the boundary layer, the dissipation integral method uses the whole distribution. This work gives an overview over dissipation integral method and extends them to three-dimensional boundary layers. The general integral equations for the three-dimensional case are derived. Using two different sets of mean velocity profiles the hyperbolical character of a dissipation integral method is shown. Apart from the integral momentum balance, the dissipation integral method satisfies a second major balance with the integral balance of mechanical energy. It is found that for a practical calculation the integral momentum equation and the integral energy equation are most useful. Sixteen two-dimensional experimental test cases with none-zero pressure gradients were computed. It was found that the averaged relative deviation between measured and computed values for the skin friction coefficient is about 5 % and about 3 % for the shape parameter. Two three-dimensional fully turbulent boundary layers approaching an obstacle where computed. The agreement between experimental results and the calculation is reasonably good. The calculation allows the prediction of the velocity distributions. Flow angle and flow gradient angle distributions being additional results

Structural and functional investigation of ABC transporter STE6-2p from Pichia pastoris reveals unexpected interaction with sterol molecules

Adenosine triphosphate (ATP)-binding cassette (ABC) transporters are multidomain transmembrane proteins, which facilitate the transport of various substances across cell membranes using energy derived from ATP hydrolysis. They are important drug targets since they mediate decreased drug susceptibility during pharmacological treatments. For the methylotrophic yeast Pichia pastoris, a model organism that is a widely used host for protein expression, the role and function of its ABC transporters is unexplored. In this work, we investigated the Pichia ABC-B transporter STE6-2p. Functional investigations revealed that STE6-2p is capable of transporting rhodamines in vivo and is active in the presence of verapamil and triazoles in vitro. A phylogenetic analysis displays homology among multidrug resistance (MDR) transporters from pathogenic fungi to human ABC-B transporters. Further, we present high-resolution single-particle electron cryomicroscopy structures of an ABC transporter from P. pastoris in the apo conformation (3.1 Å) and in complex with verapamil and adenylyl imidodiphosphate (AMP-PNP) (3.2 Å). An unknown density between transmembrane helices 4, 5, and 6 in both structures suggests the presence of a sterol-binding site of unknown function

Opportunities for Seaweed Aquaculture Development in the Azores

Jornadas "Ciência nos Açores – que futuro? Tema Ciências Naturais e Ambiente", Ponta Delgada, 7-8 de Junho de 2013.As macroalgas marinhas têm sido largamente usadas para diversos fins. Nos Açores, várias espécies têm sido usadas tradicionalmente na alimentação humana (e.g. Fucus spiralis, Porphyra spp., Laurencia spp. e Osmundea spp.) e para extracção de compostos com interesse na indústria dos ficocolóides (Pterocladiella capilacea e Gelidium spp.). As exigências no controlo da qualidade e as práticas actuais de colheita de macroalgas marinhas selvagens na Europa levantam preocupações ambientais sérias que tornam premente a necessidade se implementarem métodos de produção de biomassa controlados, como é o caso da aquacultura de macroalgas marinhas. Apesar da importância da exploração sustentável dos recursos marinhos existentes nos Açores, não existe informação sobre a viabilidade do cultivo de macroalgas marinhas no Arquipélago. O conhecimento sobre os requisitos básicos para o cultivo em grande escala das espécies nativas seleccionadas e os locais mais apropriados para a sua implementação está em falta. O objectivo principal do presente projecto é avaliar o potencial de cultivo de espécies de macroalgas marinhas seleccionadas, bem como identificar os métodos de cultivo mais adequados. Os resultados do programa de doutoramento serão de extrema importância quer em termos científicos quer em termos empresariais. Permitirão a transferência de tecnologia para o tecido empresarial regional e para a implementação de empresas de base tecnológica indo ao encontro das futuras políticas de financiamento europeias no âmbito do Programa Europeu Horizonte 2020.ABSTRACT: Seaweeds have a wide range of applications. In the Azores, several species of seaweeds were traditionally used either as food (e.g. Fucus spiralis, Porphyra spp., Laurencia spp. and Osmundea spp.) or for extraction of chemical products (Pterocladiella capilacea e Gelidium spp.). The product quality control requirements and concerns regarding the environmental sustainability of current wild seaweed biomass harvesting practices in Europe demand for controlled seaweed aquaculture. Despite the interest in exploiting Azorean seaweed resources, there is no information on the feasibility of cultivating seaweed in the Azores. Basic knowledge on large scale cultivation requirements of the selected native species is missing. The present project it’s aimed at evaluating the culture potential of selected Azorean species. The resulting outputs will be extremely important for both academic and economic purposes, bringing together the research and the market. Innovative enterprise will benefit from this project and develop technological breakthroughs into viable products with real commercial potential. This main objective is in according with the principal strategy of the Horizon 2020 that will tackle societal challenges after the end of FP7.Fundação para a Ciência e Tecnologi

Efficient microbial bioconversion of brown macroalgae obtained through profitable high-density sea cultivation using modified microbial strains to produce commodity and specialty chemicals: A developing blue chemical industry in Chile

Plant biomass is considered a promising feedstock for large scale sustainable bio-based green chemistry. However, only the use of agricultural or forestry residues is viable, since they do not compete for land with feed crops and have competitive costs. Moreover, carbohydrate recovery from these sources is always difficult due to their high lignin content. Alternatively, macroalgae are competitive sources of carbohydrate-rich biomass not requiring land or fresh water for its production. Macrocystis pyrifera is one of the fastest-growing macroalgal species with high CO2 fixation efficiency, highly-abundant and accessible carbohydrates. We demonstrated that it can be cultured in temperate seas, yielding 124 ton/Ha/yr, and can be economically profitable at a 10-hectare scale 1,2. Microbial and enzymatic algal biomass bioprocessing has been also undertaken by our group. We demonstrated the technical feasibility of producing ethanol at a pilot industrial scale by fermenting algal carbohydrates with a genetically modified Escherichia coli 3. However, ethanol production, even with high productivities, was not commercially viable. To make algal biomass bioconversion profitable, we performed a large metabolic engineering and synthetic biology project to discover combinations of metabolic pathways, regulation, carbohydrate sources –algal or not– and alternative bioproducts that maximize microbial efficiency and commercial viability. Using a genome-scale reconstruction of Saccharomyces cerevisiae’s metabolism, we demonstrated that redox ratio constraints and the preferential use of NADH or NADPH for alginate metabolism were key for S. cerevisiae conversion of alginate:mannitol carbohydrate sources 4. However, yeast use makes chemical processes technically and economically unfeasible for low value products due to their inability to produce extracellular enzymes for alginate lysis. By means of dynamic metabolic models developed for E. coli, we demonstrated that the main metabolic process bottleneck is microbial carbohydrate metabolization and that algal carbohydrate composition is a key determinant of fermentation efficiency. Using a multi-objective optimization strategy focused on microorganism growth, energy levels and redox ratio conservation, we also showed that ethanol production from algal biomass is incompatible with E. coli’s metabolism, due to low energetic and redox efficiencies obtained from alginate using host microorganism metabolic pathways. We then used high-performance parallel computing to develop a metabolic potentiality map for E. coli in which we explored more than 10.000 possible combinations of metabolic pathways that could be built in our strain to convert brown macroalgae carbohydrates with high efficiency, considering the best combinations of knock-outs and overexpressions to be introduced in E. coli’s central metabolic pathways. With this technique, we identified other valuable chemicals, such as succinic, aspartic, gluconic and levulinic acids, and complex aromatic and aliphatic biomolecules can be efficiently produced from Macrocystis with specifically modified strains for each product. The bulk of our research fostering algal feedstock production and industrial bioconversion in Chile will be presented in this work. 1. Buschmann, A. H. et al. The Status of Kelp Exploitation and Marine Agronomy, with Emphasis on Macrocystis pyrifera, in Chile. Advances in Botanical Research 71, 161–188 (2014). 2. Camus, C., Infante, J. & Buschmann, A. H. Overview of 3 year precommercial seafarming of Macrocystis pyrifera along the Chilean coast. Reviews in Aquaculture 10, 543–559 (2018). 3. Camus, C. et al. Scaling up bioethanol production from the farmed brown macroalga Macrocystis pyrifera in Chile. Biofuels, Bioproducts and Biorefining 10, 673–685 (2016). 4. Contador, C. A. et al. Analyzing redox balance in a synthetic yeast platform to improve utilization of brown macroalgae as feedstock. Metabolic Engineering Communications 2, 76–84 (2015)