Novel Approaches for Fungal Transcriptomics from Host Samples.

Candida albicans adaptation to the host requires a profound reprogramming of the fungal transcriptome as compared to in vitro laboratory conditions. A detailed knowledge of the C. albicans transcriptome during the infection process is necessary in order to understand which of the fungal genes are important for host adaptation. Such genes could be thought of as potential targets for antifungal therapy. The acquisition of the C. albicans transcriptome is, however, technically challenging due to the low proportion of fungal RNA in host tissues. Two emerging technologies were used recently to circumvent this problem. One consists of the detection of low abundance fungal RNA using capture and reporter gene probes which is followed by emission and quantification of resulting fluorescent signals (nanoString). The other is based first on the capture of fungal RNA by short biotinylated oligonucleotide baits covering the C. albicans ORFome permitting fungal RNA purification. Next, the enriched fungal RNA is amplified and subjected to RNA sequencing (RNA-seq). Here we detail these two transcriptome approaches and discuss their advantages and limitations and future perspectives in microbial transcriptomics from host material.

Romanisation in Gaul: new methodological approaches for the study of Gaulish fine wares (200 B.C.-50 A.D.).

The twenty-second Theoretical Roman Archaeology Conference (TRAC) was held at the Goethe-University Frankfurt am Main in spring 2012. During the three-day conference fifty papers were delivered, discussing issues from a wide range of geographical regions of the Roman Empire, and applying various theoretical and methodological approaches. An equally wide selection of subjects was presented: sessions looked at Greek art and philhellenism in the Roman world, the validity of the concept of 'Romanisation', change and continuity in Roman religion, urban neighbourhood relations in Pompeii and Ostia, the transformation of objects in and from the Roman world, frontier markets and Roman archaeology in the Provinces. In addition, two general sessions covered single topics such as the 'transvestite of Catterick', metal recycling or Egyptian funeral practice in the Roman period. This volume contains a selection of papers from all these sessions.

Personalized approaches to active immunotherapy in cancer

Immunotherapy is emerging as a promising anti-cancer curative modality. However, in contrast to recent advances obtained employing checkpoint blockade agents and T cell therapies, clinical efficacy of therapeutic cancer vaccines is still limited. Most vaccination attempts in the clinic represent "off-the shelf" approaches since they target common "self" tumor antigens, shared among different patients. In contrast, personalized approaches of vaccination are tailor-made for each patient and in spite being laborious, hold great potential. Recent technical advancement enabled the first steps in the clinic of personalized vaccines that target patient-specific mutated neo-antigens. Such vaccines could induce enhanced tumor-specific immune response since neo-antigens are mutation-derived antigens that can be recognized by high affinity T cells, not limited by central tolerance. Alternatively, the use of personalized vaccines based on whole autologous tumor cells, overcome the need for the identification of specific tumor antigens. Whole autologous tumor cells could be administered alone, pulsed on dendritic cells as lysate, DNA, RNA or delivered to dendritic cells in-vivo through encapsulation in nanoparticle vehicles. Such vaccines may provide a source for the full repertoire of the patient-specific tumor antigens, including its private neo-antigens. Furthermore, combining next-generation personalized vaccination with other immunotherapy modalities might be the key for achieving significant therapeutic outcome.

Estimating extreme value cumulative distribution functions using bias-corrected kernel approaches

We propose a new kernel estimation of the cumulative distribution function based on transformation and on bias reducing techniques. We derive the optimal bandwidth that minimises the asymptotic integrated mean squared error. The simulation results show that our proposed kernel estimation improves alternative approaches when the variable has an extreme value distribution with heavy tail and the sample size is small.

Convergent Approaches for the Synthesis of the Anti-tumoral Peptide, Kahalalide F. Study of Orthogonal Protecting Groups

Kahalalide compounds are peptides that are isolated from a Hawaiian herbivorous marine species of mollusc, Elysia rufescens, and its diet, the green alga Bryopsis sp. Kahalalide F and its synthetic analogues are the most promising compounds of the Kahalalide family because they show anti-tumoral activity. Linear solid-phase syntheses of Kahalalide F have been reported. Here we describe several new improved synthetic routes based on convergent approaches with distinct orthogonal protection schemes for the preparation of Kahaladide analogues. These strategies allow a better control and characterization of the intermediates because more reactions are performed in solution. Five derivatives of Kahalalide F were synthesized using several convergent approaches.

Spiders on a Hot Volcanic Roof: Colonisation Pathways and Phylogeography of the Canary Islands Endemic Trap-Door Spider Titanidiops canariensis (Araneae, Idiopidae)

Studies conducted on volcanic islands have greatly contributed to our current understanding of how organisms diversify. The Canary Islands archipelago, located northwest of the coast of northern Africa, harbours a large number of endemic taxa. Because of their low vagility, mygalomorph spiders are usually absent from oceanic islands. The spider Titanidiops canariensis, which inhabits the easternmost islands of the archipelago, constitutes an exception to this rule. Here, we use a multi-locus approach that combines three mitochondrial and four nuclear genes to investigate the origins and phylogeography of this remarkable trap-door spider. We provide a timeframe for the colonisation of the Canary Islands using two alternative approaches: concatenation and species tree inference in a Bayesian relaxed clock framework. Additionally, we investigate the existence of cryptic species on the islands by means of a Bayesian multi-locus species delimitation method. Our results indicate that T. canariensis colonised the Canary Islands once, most likely during the Miocene, although discrepancies between the timeframes from different approaches make the exact timing uncertain. A complex evolutionary history for the species in the archipelago is revealed, which involves two independent colonisations of Fuerteventura from the ancestral range of T. canariensis in northern Lanzarote and a possible back colonisation of southern Lanzarote. The data further corroborate a previously proposed volcanic refugium, highlighting the impact of the dynamic volcanic history of the island on the phylogeographic patterns of the endemic taxa. T. canariensis includes at least two different species, one inhabiting the Jandia peninsula and central Fuerteventura and one spanning from central Fuerteventura to Lanzarote. Our data suggest that the extant northern African Titanidiops lineages may have expanded to the region after the islands were colonised and, hence, are not the source of colonisation. In addition, T. maroccanus may harbour several cryptic species.

Synthetic approaches towards the Lycopodium alkaloids

The Lycopodium alkaloids are a structurally diverse group of natural products isolated from Lycopodium with important biological effects for the potential treatment of cancer and severe neurodegenerative diseases. To date, full biological studies have been hampered by lack of material from natural sources. Total synthesis represents a possible solution to meet this demand as well as the most effective way to design new compounds to determine structural activity relationships and obtain more potent compounds. The aim of this chapter is to summarise the work carried out in this field so far by presenting an overview of the synthetic strategies used to access each of the four key Lycopodium alkaloid types. Particular emphasis has been placed on methods that rapidly construct each nucleus utilizing tandem reactions.

Synthetic approaches towards the Lycopodium alkaloids

The Lycopodium alkaloids are a structurally diverse group of natural products isolated from Lycopodium with important biological effects for the potential treatment of cancer and severe neurodegenerative diseases. To date, full biological studies have been hampered by lack of material from natural sources. Total synthesis represents a possible solution to meet this demand as well as the most effective way to design new compounds to determine structural activity relationships and obtain more potent compounds. The aim of this chapter is to summarise the work carried out in this field so far by presenting an overview of the synthetic strategies used to access each of the four key Lycopodium alkaloid types. Particular emphasis has been placed on methods that rapidly construct each nucleus utilizing tandem reactions.

Late Roman amphorae in the eastern Tarraconensis. Some chronological and quantitative approaches

Since the classic study of Simon J. Keay published in 1984, knowledge of late Roman amphorae has progressed markedly, thanks to scholars such as Michel Bonifay and Paul Reynolds, amongst others. The area studied by Keay was Catalonia, the ancient Eastern Tarraconensis. The overview here offered for this same region reveals the central role played by African imports in late Antique times, with a minor presence of the Eastern-Mediterranean and South-Hispanic (both Baetican and Lusitanian) productions. Progress in research in the last 25 years has been centred on a series of new and well-dated contexts: the data they have yielded has clarified more precisely the chronology and the proportions of the different imports. On occasion a quantitative approach may even be applied. At the same time the relationship between town and country with respect to the late Roman amphorae is proving of interest and providing results of significance.

Advanced chemometric methods: Applicability on industrial data

Recent years have produced great advances in the instrumentation technology. The amount of available data has been increasing due to the simplicity, speed and accuracy of current spectroscopic instruments. Most of these data are, however, meaningless without a proper analysis. This has been one of the reasons for the overgrowing success of multivariate handling of such data. Industrial data is commonly not designed data; in other words, there is no exact experimental design, but rather the data have been collected as a routine procedure during an industrial process. This makes certain demands on the multivariate modeling, as the selection of samples and variables can have an enormous effect. Common approaches in the modeling of industrial data are PCA (principal component analysis) and PLS (projection to latent structures or partial least squares) but there are also other methods that should be considered. The more advanced methods include multi block modeling and nonlinear modeling. In this thesis it is shown that the results of data analysis vary according to the modeling approach used, thus making the selection of the modeling approach dependent on the purpose of the model. If the model is intended to provide accurate predictions, the approach should be different than in the case where the purpose of modeling is mostly to obtain information about the variables and the process. For industrial applicability it is essential that the methods are robust and sufficiently simple to apply. In this way the methods and the results can be compared and an approach selected that is suitable for the intended purpose. Differences in data analysis methods are compared with data from different fields of industry in this thesis. In the first two papers, the multi block method is considered for data originating from the oil and fertilizer industries. The results are compared to those from PLS and priority PLS. The third paper considers applicability of multivariate models to process control for a reactive crystallization process. In the fourth paper, nonlinear modeling is examined with a data set from the oil industry. The response has a nonlinear relation to the descriptor matrix, and the results are compared between linear modeling, polynomial PLS and nonlinear modeling using nonlinear score vectors.

Lo que no es nuevo puede ser actual: hacia distintos puntos de mira de la didáctica del arte What is not new can be current - towards different approaches of the didactics of art

With this article we want to present a state of affairs of the didactics of art in our context, and at the same time we want todiscuss on its pluri- and interdisciplinary construction. We check the different disciplines that configure it and we analysethe paradigm of artistic education as a discipline (DBAE) and its passage to post-modernity. This example focuses thediscussion about the opportunity of adapting holistic educative models and the transition of the current educativeinnovation towards skill models

Carbon sequestration rates in Swedish forest soils : a comparison of three approaches

Selective papers of the workshop on "Development of models and forest soil surveys for monitoring of soil carbon", Koli, Finland, April 5-9 2006.

Delineamentos experimentais eficientes para estudos de cinética química

In this paper we show how to obtain efficient designs of experiments for fitting Michaelis-Menten and Hill equations useful in chemical studies. The search of exact D-optimal designs by using local and pseudo-Bayesian approaches is considered. Optimal designs were compared to those commonly used in practice using an efficiency measure and theoretical standard errors of the kinetic parameter estimates. In conclusion, the D-optimal designs based on the Hill equation proved efficient for estimating the parameters of both models. Furthermore, these are promising with respect to practical issues, allowing efficient estimation as well as goodness-of-fit tests and comparisons between some kinetic models.

Developing a Comprehensive Model for Marketing Planning — Combining the Major Approaches

Mix marketing and relationships marketing are two major approaches that often form a basis for organizational marketing planning. The superiority of these approaches has been debated for long without any rational conclusion. Lately there have been studies indicating that both of the major approaches are many times used side by side in marketing planning. There have been also studies suggesting that even combining the mix marketing and relationship marketing approaches might be possible. The aim of this thesis is to provide knowledge about the usage of mix marketing and relationship marketing approaches in organizations and possibilities in combining the approaches. Also a settlement of strengths, weaknesses and risks of combining is intended to provide. The objectives were met through the literature and a case study research. In the case study, interviews were conducted in order to gain a deeper knowledge about marketing planning in various organizations. Based on this study, the combining of the major marketing approaches will be possible and even recommended when keeping in mind few aspects which might cause some troubles in the combining process.

Lipase-catalyzed Approaches towards Secondary Alcohols: Intermediates for Enantiopure Drugs

The use of enantiopure intermediates for drug synthesis is a trend in pharmaceutical industry. Different physiological effects are associated with the enantiomers of chiral molecules. Thus, the safety profile of a drug based on an enantiopure active pharmaceutical ingredient is more reliable. Biocatalysis is an important tool to access enantiopure molecules. In biocatalysis, the advantage of selectivity (chemo-, regio- and stereoselectivity) is combined with the benefits of a green synthesis strategy. Chemoenzymatic syntheses of drug molecules, obtained by combining biocatalysis with modern chemical synthesis steps usually consists of fewer reaction steps, reduced waste production and improved overall synthetic efficiency both in yields and enantio- and/or diastereoselectivities compared with classical chemical synthesis. The experimental work together with the literature review clearly indicates that lipase catalysis is highly applicable in the synthesis of enantiopure intermediates of drug molecules as the basis to infer the correct stereochemistry. By lipase catalysis, enantiopure secondary alcohols used as intermediates in the synthesis of Dorzolamide, an antiglaucoma drug, were obtained. Enantiopure _-hydroxy nitriles as potential intermediates for the synthesis of antidepressant drugs with 1-aryl-3- methylaminopropan-1-ol structure were also obtained with lipases. Kinetic resolution of racemates was the main biocatalytic approach applied. Candida Antarctica lipase B, Burkholderia cepacia lipase and Thermomyces lanuginosus lipase were applied for the acylation of alcohols and the alcoholysis of their esters in organic solvents, such as in diisopropyl ether and tert-butyl methyl ether. Candida Antarctica lipase B was used under solvent free conditions for the acylation of ethyl 3-hydroxybutanoate.