Fermentation of fruit juices by the osmotolerant yeast Candida magnoliae

This study focuses on the assessment of the fermentation conditions required to modulate the metabolic flux in the osmotolerant yeast Candida magnoliae and evaluate its potential to produce low-alcoholic and low-caloric fermented beverages. For that purpose, two strains, PYCC 2903 and PYCC 3191, were used and fermentation conditions as oxygenation, sugar concentration and the ratio of glucose to fructose were studied using synthetic culture media. Candida magnoliae PYCC 2903 was subsequently used to ferment real industrial fructose-rich substrates such as fruit juices. Sugar consumption profiles for C.magnoliae PYCC 2903 incubated aerobically in the presence of high fructose and glucose concentrations (15%, 10% and 5%) showed a selective utilization of fructose, denoting a preference for this sugar over glucose. The lower ratio between ethanol and sugar alcohols yield was obtained for both strains incubated under oxygen limitation simulating industrial fructose-rich substrates, confirming the ability of this yeast to direct fermentation towards alternative products. Enzymatic assays for hexokinase activity in terms of capacity and affinity for glucose and fructose were performed, aiming to elucidate its contribution to the fructophilic behaviour of this yeast. Enzymatic assays for both strains showed that the Vmax is two to threefold higher for fructose than for glucose but Km is also 10-20-fold higher for this sugar than for glucose. Hence, hexokinase kinetic properties do not explain fructophily in C.magnoliae. This indicates that fructose transport is probably determining in this respect, as observed for other fructophilic yeasts. Fruit juice fermentations with C.magnoliae PYCC 2903 revealed a potential for the production of beverages with interesting sensorial properties. Pear and peach fermentations exhibited the best results with the lowest ratio between ethanol and sugar alcohols yield and the most pleasant organoleptic features.

Contribution to drug discovery and development for tauopathies using yeast as a model

This work aimed to contribute to drug discovery and development (DDD) for tauopathies, while expanding our knowledge on this group of neurodegenerative disorders, including Alzheimer’s disease (AD). Using yeast, a recognized model for neurodegeneration studies, useful models were produced for the study of tau interaction with beta-amyloid (Aβ), both AD hallmark proteins. The characterization of these models suggests that these proteins co-localize and that Aβ1-42, which is toxic to yeast, is involved in tau40 phosphorylation (Ser396/404) via the GSK-3β yeast orthologue, whereas tau seems to facilitate Aβ1-42 oligomerization. The mapping of tau’s interactome in yeast, achieved with a tau toxicity enhancer screen using the yeast deletion collection, provided a novel framework, composed of 31 genes, to identify new mechanisms associated with tau pathology, as well as to identify new drug targets or biomarkers. This genomic screen also allowed to select the yeast strain mir1Δ-tau40 for development of a new GPSD2TM drug discovery screening system. A library of unique 138 marine bacteria extracts, obtained from the Mid-Atlantic Ridge hydrothermal vents, was screened with mir1Δ-tau40. Three extracts were identified as suppressors of tau toxicity and constitute good starting points for DDD programs. mir1Δ strain was sensitive to tau toxicity, relating tau pathology with mitochondrial function. SLC25A3, the human homologue of MIR1, codes for the mitochondrial phosphate carrier protein (PiC). Resorting to iRNA, SLC25A3 expression was silenced in human neuroglioma cells, as a first step towards the engineering of a neural model for replicating the results obtained in yeast. This model is essential to understand the mechanisms of tau toxicity at the mitochondrial level and to validate PiC as a relevant drug target. The set of DDD tools here presented will foster the development of innovative and efficacious therapies, urgently needed to cope with tau-related disorders of high human and social-economic impact.

The expression in saccharomyces cerevisiae of a glucose/xylose symporter from Candida intermedia is affected by the presence of a glucose/xylose facilitator

Microbiology, 154

Biogeography, host specificity, and molecular phylogeny of the basidiomycetous yeast phaffia rhodozyma and its sexual form, xanthophyllomyces dendrorhous

Applied and Environmental Microbiology, Vol. 73, No.4

Identification of mating type genes in the bipolar basidiomycetous yeast rhodosporidium toruloides: first insight into the MAT Locus structure of the sporidiobolales

Eukaryotic Cell, Vol.7, Nº6

Farysizyma gen. nov., an anamorphic genus in the Ustilaginales to accommodate three novel epiphytic basidiomycetous yeast species from America, Europe and Asia

FEMS Yeast Research, Vol. 8, Nº 3

Paleoclimatic correlations between Western european continental and marine areas Contribution of big villafranchian fauna

It is very difficult to make paleoclimatic correlations between continental and marine areas, but it is possible with biostratigraphic data. Reliable correlations can be made only between broad periods: between 3.5 and 3 Ma, around 2.4 Ma, until 1.6 Ma and after 1.6 Ma. The arid Mediterranean phases led to the disappearance of the European Villafranchian fauna (1.0 Ma).

The marine Neogene of Eastern Venezuela : a preliminary report

Proceedings of tile 1" R.C.A.N.S. Congress, Lisboa, October 1992

The Cristo Rei section (Lower Miocene). Distal fluviatile environments in a marine series, plants, vertebrates and other evidence, age

The section at Cristo Rei shows sandy beds with intercalated clayey lenses (IVb division from the Lisbon Miocene series) that correspond to a major regression event dated from between ca. 17.6 and 17 Ma. They also correspond to a distal position (relatively to the typical fluviatile facies in Lisbon), nearer the basin's axis. Geologic data and paleontological analysis (plant fossils, fishes, crocodilians, land mammals) allow the reconstruction of environments that were represented in the concerned area: estuary with channels and ox-bows; upstream, areas occupied by brackish waters where Gryphaea griphoides banks developped; still farther upstream, freshwaters sided by humid forests and low mountain subtropical forests under warm temperate and rainy conditions, as well as not far away, seasonally dry environments (low density tree or shrub cover, or steppe).

The three main marine depositional cycles of the Neogene of Portugal

The present work follows a stratigraphic model for the marine Neogene of Portugal based on the definition of three main marine sedimentary cycles. Conceptually the I, II and III Neogene Cycles can be defined as 2nd order sedimentary sequences with duration ranging from 5 to 8 Ma. The I Neogene Cycle is fully represented only in the Lower Tagus Basin. Ranging from the Early Aquitanian to the Late Burdigalian the I Neogene Cycle testify a transgressive episode in the region of Lisbon and Setúbal Peninsula. Rapid lateral facies variations suggest a shallowmarine basin. This cycle ends with an important Late Burdigalian tectonic compressive event expressed by uplift of the surrounding areas and deformation affecting the Early Miocene deposits of the Arrábida Chain. The II Neogene Cycle includes thick sedimentary sequences covering Paleozoic and Mesozoic formations in the Algarve and Alvalade-Melides regions and it extends as far north as Santarém in the Lower Tagus Basin. Mainly controlled by global eustasy, it was generated by the important positive eustatic trend that characterized the Middle Miocene worldwide to which the Portuguese continental margin acted more or less passively. This cycle ended with a second and the most important compression event starting after the end of the Serravallian affecting the entire Portuguese onshore and shelf areas. This led to an important depositional hiatus of marine sediments for more than 2.5 Ma. During the Early and the Middle Tortonian occurred the clockwise rotation of the Guadalquivir Basin. The thickmarine units deposited afterwards in this basin produced a litostatic load, which seems to have induced subsidence farther west resuming the Neogene marine sedimentation in the Cacela region (Eastern Algarve), during the Late Tortonian. This marks the beginning of the III Neogene Cycle. To the north, in the Sado Basin (Alvalade-Melides region), a similar depositional sequence starts its sedimentation during the Messinian. Further north, in the Pombal-Caldas da Rainha region, marine sedimentation started during the Late Pliocene (Piacenzian). The migration in time, from south to north for the beginning of the marine sedimentation of this cycle is interpreted as reflecting a visco-elastic propagation of the deformation from the Betic chain northwards.

Late Quaternary warm marine mollusks from Santa Maria (Azores); paleoecologic and paleobiogeographic considerations

The sub-fossil fauna from the Late Quaternary marine deposits of Santa Maria is made of more than 50 species of gastropods and bivalves, 19 of them collected recently and for the first time in the northern coast of the island (Lagoinhas Bay). The sub-fossil shells are found in deposits of beach sands, situated 2-3 meters above the present low tide. The carbonated sands from the basal part of the succession yield an autochthonous association of borers dominated by the bivalve Myoforceps aristata (Dillwin, 1817). Upwards, the marine sands contain concentrations of beach drift shells, including well-preserved supratidal and intertidal gastropods, among them a large number of Rissoidae. The bivalve fauna is dominated by disarticulated valves of Ervilia castanea (Montagu, 1803), a small infaunal coloniser of mobile sandy substrates. The composition of the fauna is made essentially of West European species, many of them common to the West Coast of Portugal. However, a few "warm guests" with West African or Caribbean affinities were also found, suggesting a close relation with some of the "Tyrrhenian" warm associations found in the Western Mediterranean.

Miocene marine mammals from Portugal. Paleogeographical and paleoecological significance

Neogene marine mammals are still incompletely known in Portugal. However, a general overview of the geographic and stratigraphic distribution of marine mammal localities in the Miocene of Portugal is already possible. An attempt of correlation between the trends shown by these distributions and the horizontal and vertical environmental shifts is presented. In general, sirenians occur in deposits representing shallow, warm, low energy aquatic environments; while cetaceans are more frequent in more open, deep and temperate marine environments.

Miocene marine-continental correlations in the Lisbon area and some discussion related to personnal experience

Sandpit exploitation near Lisbon allowed collecting of many Miocene, non marine fossils. These sands are part of the mostly marine Miocene series in the Lower Tagus basin. The particularly favourable situation led several researchers to deal with marine-continental correlations. Difficulties often concern methodologic aspects. Some poorly based interpretations exerced a lasting influence. A critical approach is presented. Analysis requires data. Methods based upon models often lead to the temptation of fitting data in order to confirm a priori conclusions, or of mixing up data as if of equal statistic value while they have not at all the same weight. Erroneous interpretations' uncritical repetition for many years "upgraded" them into absolute truth. Another point is endemism vs. europeism. Miocene mammals from Lisbon compared well with corresponding French, contemporaneous taxa, while this was apparently not true for Spanish ones. Too much accent had been put on the endemic character of Spanish, or even regional, mammalian faunas. Nationalist bias and sensationalism also weigh, albeit negatively. Meanwhile nearly all the more evident examples as the rhinoceros Hispanotherium are discredited as Iberian endemisms. Taxa may appear as endemic just because they have not yet been found elsewhere. At least for the medium to large-sized mammals, with their huge geographic distribution, faunal differences depend much more on ecology, climate and environmental conditions. Emphasis on differences may also result from researchers that are often in a precarious situation and need very much to achieve short-term, preferably sensational results. Overvalued differences may mask real similarities. Unethic and not scientific behaviour are further enhanced by "nomina nuda" tricks that may simply be a way to circunvent or cheat the Priority Rule. On the other hand, access to communication networks may present as sensational novelties items that are not new at all, misleading the audience. A new class of "science people" arose, created by the media and not by the value of their real achievements. Discussion is presented on sedimentation processes and discontinuities that are often regarded as absolute precision dating tools, as well as on some geochemical and paleomagnetic interpretations. A very good chronologie frame has been obtained for the basin under study on the basis of an impressive set of data, providing a rather detailed and accurate frame for Miocene marine-continental correlations.

Communication and choice in yeast mating

Dissertação apresentada para obtenção do grau de doutor em Biologia de Sistemas pelo Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa.