Gravitational microscope in MACSJ1206: unveiling galaxy cluster member mass distribution

Gravitational lensing is a powerful tool to investigate the properties of the distribution of matter, be it barionic or dark. In this work we take advantage of Strong Gravitational Lensing to infer the properties of one of the galaxy-scale substructures that makes up the cluster MACSJ1206. It is relatively easy to model the morphology of the visible components of a galaxy, while the morphology of the dark matter distribution cannot be so easily constrained. Being sensitive to the whole mass, strong lensing provides a way to probe DM distribution, and this is the reason why it is the best tool to study the substructure. The goal of this work consists of performing an analysis of the substructure previously mentioned, an early type galaxy (ETG), by analyzing the highly magnified Einstein ring around it, in order to put stringent constraints on its matter distribution, that, for an ETG, is commonly well described by an isothermal profilele. This turns out to be interesting for three main different reasons. It is well known that galaxies in clusters are subject to interaction processes, both dynamic and hydrodynamic, that can significantly modify the distribution of matter within them. Therefore, finding a different profile from the one usually expected could be a sign that the galaxy has undergone processes that have changed its structure. Studying the mass distribution also means studying the dark matter component, which not only still presents great questions today, but which is also not obviously distributed in the same way as in an isolated galaxy. What emerges from the analysis is that the total mass distribution of the galaxy under examination turns out to have a slope much steeper than the isothermal usually expected.

Variations on a theme: diversification of cuticular hydrocarbons in a clade of cactophilic Drosophila

Background: We characterized variation and chemical composition of epicuticular hydrocarbons (CHCs) in the seven species of the Drosophila buzzatii cluster with gas chromatography/mass spectrometry. Despite the critical role of CHCs in providing resistance to desiccation and involvement in communication, such as courtship behavior, mating, and aggregation, few studies have investigated how CHC profiles evolve within and between species in a phylogenetic context. We analyzed quantitative differences in CHC profiles in populations of the D. buzzatii species cluster in order to assess the concordance of CHC differentiation with species divergence. Results: Thirty-six CHC components were scored in single fly extracts with carbon chain lengths ranging from C(29) to C(39), including methyl-branched alkanes, n alkenes, and alkadienes. Multivariate analysis of variance revealed that CHC amounts were significantly different among all species and canonical discriminant function (CDF) analysis resolved all species into distinct, non-overlapping groups. Significant intraspecific variation was found in different populations of D. serido suggesting that this taxon is comprised of at least two species. We summarized CHC variation using CDF analysis and mapped the first five CHC canonical variates (CVs) onto an independently derived period (per) gene + chromosome inversion + mtDNA COI gene for each sex. We found that the COI sequences were not phylogenetically informative due to introgression between some species, so only per + inversion data were used. Positive phylogenetic signal was observed mainly for CV1 when parsimony methods and the test for serial independence (TFSI) were used. These results changed when no outgroup species were included in the analysis and phylogenetic signal was then observed for female CV3 and/or CV4 and male CV4 and CV5. Finally, removal of divergent populations of D. serido significantly increased the amount of phylogenetic signal as up to four out of five CVs then displayed positive phylogenetic signal. Conclusions: CHCs were conserved among species while quantitative differences in CHC profiles between populations and species were statistically significant. Most CHCs were species-, population-, and sex-specific. Mapping CHCs onto an independently derived phylogeny revealed that a significant portion of CHC variation was explained by species' systematic affinities indicating phylogenetic conservatism in the evolution of these hydrocarbon arrays, presumptive waterproofing compounds and courtship signals as in many other drosophilid species.

Azorean Agriculture Efficiency by PAR

Copyright © 2013 Springer Netherlands.

On clustering interval data with different scales of measures : experimental results

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Clustering an interval data set : are the main partitions similar to a priori partition?

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Fluorescence in nanostructured fulleride films

Applied Physics Letters, 89

Diversidade de culturas organizacionais: obstáculos e desafios para a liderança de um mega agrupamento

Este projeto tem por base um problema identificado num mega agrupamento de um concelho do grande porto, criado em junho de 2012, e que se centra nas dificuldades de gestão e liderança de uma organização resultante da agregação de duas unidades com culturas organizacionais distintas. Na perspetiva de conhecermos e compreendermos o problema e respondermos ao nosso questionamento, optamos por uma metodologia qualitativa seguindo o método de um estudo de caso. No estudo empírico, cuja técnica de recolha foi um focus group, recolheram-se as perceções dos professores das unidades agregadas sobre a criação da nova organização, sobre o impacto nas identidades e culturas organizacionais, sobre a forma da procura de uma coesão no respeito pela diversidade e identificaram-se os quadros de referência das escolas que se agregaram. O estudo realizado permite reconhecer que as dificuldades ao nível da gestão e liderança se confinam mais na escola que perdeu a centralidade e que se sente numa posição periférica. Na elaboração do plano de ação recuperámos e valorizámos a ideia de que um modelo de liderança baseado em valores poderia ser um caminho para a unidade. Neste sentido o plano de ação, com base em alguns princípios de atuação e definição de estratégias tenta dar resposta à questão de partida e em congruência com um modelo de liderança por valores.

Host sex and ectoparasites choice: preference for, and higher survival on female hosts.

1. Sex differences in levels of parasite infection are a common rule in a wide range of mammals, with males usually more susceptible than females. Sex-specific exposure to parasites, e.g. mediated through distinct modes of social aggregation between and within genders, as well as negative relationships between androgen levels and immune defences are thought to play a major role in this pattern. 2. Reproductive female bats live in close association within clusters at maternity roosts, whereas nonbreeding females and males generally occupy solitary roosts. Bats represent therefore an ideal model to study the consequences of sex-specific social and spatial aggregation on parasites' infection strategies. 3. We first compared prevalence and parasite intensities in a host-parasite system comprising closely related species of ectoparasitic mites (Spinturnix spp.) and their hosts, five European bat species. We then compared the level of parasitism between juvenile males and females in mixed colonies of greater and lesser mouse-eared bats Myotis myotis and M. blythii. Prevalence was higher in adult females than in adult males stemming from colonial aggregations in all five studied species. Parasite intensity was significantly higher in females in three of the five species studied. No difference in prevalence and mite numbers was found between male and female juveniles in colonial roosts. 4. To assess whether observed sex-biased parasitism results from differences in host exposure only, or, alternatively, from an active, selected choice made by the parasite, we performed lab experiments on short-term preferences and long-term survival of parasites on male and female Myotis daubentoni. When confronted with adult males and females, parasites preferentially selected female hosts, whereas no choice differences were observed between adult females and subadult males. Finally, we found significantly higher parasite survival on adult females compared with adult males. 5. Our study shows that social and spatial aggregation favours sex-biased parasitism that could be a mere consequence of an active and adaptive parasite choice for the more profitable host.

Characterization of Fas (Apo-1, CD95)-Fas ligand interaction.

The death-inducing receptor Fas is activated when cross-linked by the type II membrane protein Fas ligand (FasL). When human soluble FasL (sFasL, containing the extracellular portion) was expressed in human embryo kidney 293 cells, the three N-linked glycans of each FasL monomer were found to be essential for efficient secretion. Based on the structure of the closely related lymphotoxin alpha-tumor necrosis factor receptor I complex, a molecular model of the FasL homotrimer bound to three Fas molecules was generated using knowledge-based protein modeling methods. Point mutations of amino acid residues predicted to affect the receptor-ligand interaction were introduced at three sites. The F275L mutant, mimicking the loss of function murine gld mutation, exhibited a high propensity for aggregation and was unable to bind to Fas. Mutants P206R, P206D, and P206F displayed reduced cytotoxicity toward Fas-positive cells with a concomitant decrease in the binding affinity for the recombinant Fas-immunoglobulin Fc fusion proteins. Although the cytotoxic activity of mutant Y218D was unaltered, mutant Y218R was inactive, correlating with the prediction that Tyr-218 of FasL interacts with a cluster of three basic amino acid side chains of Fas. Interestingly, mutant Y218F could induce apoptosis in murine, but not human cells.

The Armc10/SVH gene: Genome context, regulation of mitochondrial dynamics and protection against Aβ-induced mitochondrial fragmentation

Mitochondrial function and dynamics are essential for neurotransmission, neural function and neuronal viability. Recently, we showed that the eutherian-specific Armcx gene cluster (Armcx1-6 genes), located in the X chromosome, encodes for a new family of proteins that localise to mitochondria, regulating mitochondrial trafficking. The Armcx gene cluster evolved by retrotransposition of the Armc10 gene mRNA, which is present in all vertebrates and is considered to be the ancestor gene. Here we investigate the genomic organisation, mitochondrial functions and putative neuroprotective role of the Armc10 ancestor gene. The genomic context of the Armc10 locus shows considerable syntenic conservation among vertebrates, and sequence comparisons and CHIP-data suggest the presence of at least three conserved enhancers. We also show that the Armc10 protein localises to mitochondria and that it is highly expressed in the brain. Furthermore, we show that Armc10 levels regulate mitochondrial trafficking in neurons, but not mitochondrial aggregation, by controlling the number of moving mitochondria. We further demonstrate that the Armc10 protein interacts with the KIF5/Miro1-2/Trak2 trafficking complex. Finally, we show that overexpression of Armc10 in neurons prevents A beta-induced mitochondrial fission and neuronal death. Our data suggest both conserved and differential roles of the Armc10/Armcx gene family in regulating mitochondrial dynamics in neurons, and underscore a protective effect of the Armc10 gene against A beta-induced toxicity. Overall, our findings support a further degree of regulation of mitochondrial dynamics in the brain of more evolved mammals.

The Armc10/SVH gene: Genome context, regulation of mitochondrial dynamics and protection against Aβ-induced mitochondrial fragmentation

Mitochondrial function and dynamics are essential for neurotransmission, neural function and neuronal viability. Recently, we showed that the eutherian-specific Armcx gene cluster (Armcx1-6 genes), located in the X chromosome, encodes for a new family of proteins that localise to mitochondria, regulating mitochondrial trafficking. The Armcx gene cluster evolved by retrotransposition of the Armc10 gene mRNA, which is present in all vertebrates and is considered to be the ancestor gene. Here we investigate the genomic organisation, mitochondrial functions and putative neuroprotective role of the Armc10 ancestor gene. The genomic context of the Armc10 locus shows considerable syntenic conservation among vertebrates, and sequence comparisons and CHIP-data suggest the presence of at least three conserved enhancers. We also show that the Armc10 protein localises to mitochondria and that it is highly expressed in the brain. Furthermore, we show that Armc10 levels regulate mitochondrial trafficking in neurons, but not mitochondrial aggregation, by controlling the number of moving mitochondria. We further demonstrate that the Armc10 protein interacts with the KIF5/Miro1-2/Trak2 trafficking complex. Finally, we show that overexpression of Armc10 in neurons prevents A beta-induced mitochondrial fission and neuronal death. Our data suggest both conserved and differential roles of the Armc10/Armcx gene family in regulating mitochondrial dynamics in neurons, and underscore a protective effect of the Armc10 gene against A beta-induced toxicity. Overall, our findings support a further degree of regulation of mitochondrial dynamics in the brain of more evolved mammals.

Role and regulatory mechanisms of heat shock factor 2

Protein homeostasis is essential for cells to prosper and survive. Various forms of stress, such as elevated temperatures, oxidative stress, heavy metals or bacterial infections cause protein damage, which might lead to improper folding and formation of toxic protein aggregates. Protein aggregation is associated with serious pathological conditions such as Alzheimer’s and Huntington’s disease. The heat shock response is a defense mechanism that protects the cell against protein-damaging stress. Its ancient origin and high conservation among eukaryotes suggest that the response is crucial for survival. The main regulator of the heat shock response is the transcription factor heat shock factor 1 (HSF1), which induces transcription of genes encoding protective molecular chaperones. In vertebrates, a family of four HSFs exists (HSF1-4), with versatile functions not only in coping with acute stress, but also in development, longevity and cancer. Thus, knowledge of the HSFs will aid in our understanding on how cells survive suboptimal circumstances, but will also provide insights into normal physiological processes as well as diseaseassociated conditions. In this study, the function and regulation of HSF2 have been investigated. Earlier gene inactivation experiments in mice have revealed roles for HSF2 in development, particularly in corticogenesis and spermatogenesis. Here, we demonstrate that HSF2 holds a role also in the heat shock response and influences stress-induced expression of heat shock proteins. Intriguingly, DNA-binding activity of HSF2 upon stress was dependent on the presence of intact HSF1, suggesting functional interplay between HSF1 and HSF2. The underlying mechanism for this phenomenon could be configuration of heterotrimers between the two factors, a possibility that was experimentally verified. By changing the levels of HSF2, the expression of HSF1-HSF2 heterotrimer target genes was altered, implementing HSF2 as a modulator of HSF-mediated transcription. The results further indicate that HSF2 activity is dependent on its concentration, which led us to ask the question of how accurate HSF2 levels are achieved. Using mouse spermatogenesis as a model system, HSF2 was found to be under direct control of miR-18, a miRNA belonging to the miR-17~92 cluster/Oncomir-1 and whose physiological function had remained unclear. Investigations on spermatogenesis are severely hampered by the lack of cell systems that would mimic the complex differentiation processes that constitute male germ cell development. Therefore, to verify that HSF2 is regulated by miR-18 in spermatogenesis, a novel method named T-GIST (Transfection of Germ cells in Intact Seminiferous Tubules) was developed. Employing this method, the functional consequences of miR-18-mediated regulation in vivo were demonstrated; inhibition of miR- 18 led to increased expression of HSF2 and altered the expression of HSF2 target genes Ssty2 and Speer4a. Consequently, the results link miR-18 to HSF2-mediated processes such as germ cell maturation and quality control and provide miR-18 with a physiological role in gene expression during spermatogenesis.Taken together, this study presents compelling evidence that HSF2 is a transcriptional regulator in the heat shock response and establishes the concept of physical interplay between HSF2 and HSF1 and functional consequences thereof. This is also the first study describing miRNA-mediated regulation of an HSF.

A prospective study of hepatitis B virus markers in patients with chronic HBV infection from Brazilian families of Western and Asian origin

The purpose of the present study was to determine the frequency of hepatitis B virus (HBV) markers in families of HBsAg-positive patients with chronic liver disease. Serum anti-HBc, HBsAg and anti-HBs were determined by enzyme immunoassay and four subpopulations were considered: genetically related (consanguineous) and non-genetically related (non-consanguineous) Asian subjects and genetically related and non-genetically related Western subjects. A total of 165 and 186 relatives of Asian and Western origin were enrolled, respectively. The occurrence of HBsAg and anti-HBs antibodies was significantly higher (P < 0.0001) in family members of Asian origin (81.8%) than in family members of Western origin (36.5%). HBsAg was also more frequent among brothers (79.6 vs 8.5%; P < 0.0001), children (37.9 vs 3.3%; P < 0.0001) and other family members (33.9 vs 16.7%; P < 0.0007) of Asian than Western origin, respectivelly. No difference between groups was found for anti-HBs, which was more frequently observed in fathers, spouses and other non-genetic relatives. HBV infection was significantly higher in children of Asian than Western mothers (P < 0.0004). In both ethnic groups, the mothers contributed more to their children's infection than the fathers (P < 0.0001). Furthermore, HBsAg was more frequent among consanguineous members and anti-HBs among non-consanguineous members. These results suggest the occurrence of vertical transmission of HBV among consanguineous members and probably horizontal sexual transmission among non-consanguineous members of a family cluster. Thus, the high occurrence of dissemination of HBV infection characterizes family members as a high-risk group that calls for immunoprophylaxis. Finally, the study showed a high familial aggregation rate for both ethnic groups, 18/19 (94.7%) and 23/26 (88.5%) of the Asian and Western origin, respectively.

Nuclear organisation and epigenetic regulation of gene expression in Dictyostelium discoideum

A series of vectors for the over-expression of tagged proteins in Dictyostelium were designed, constructed and tested. These vectors allow the addition of an N- or C-terminal tag (GFP, RFP, 3xFLAG, 3xHA, 6xMYC and TAP) with an optimized polylinker sequence and no additional amino acid residues at the N or C terminus. Different selectable markers (Blasticidin and gentamicin) are available as well as an extra chromosomal version; these allow copy number and thus expression level to be controlled, as well as allowing for more options with regard to complementation, co- and super-transformation. Finally, the vectors share standardized cloning sites, allowing a gene of interest to be easily transfered between the different versions of the vectors as experimental requirements evolve. The organisation and dynamics of the Dictyostelium nucleus during the cell cycle was investigated. The centromeric histone H3 (CenH3) variant serves to target the kinetochore to the centromeres and thus ensures correct chromosome segregation during mitosis and meiosis. A number of Dictyostelium histone H3-domain containing proteins as GFP-tagged fusions were expressed and it was found that one of them functions as CenH3 in this species. Like CenH3 from some other species, Dictyostelium CenH3 has an extended N-terminal domain with no similarity to any other known proteins. The targeting domain, comprising α-helix 2 and loop 1 of the histone fold is required for targeting CenH3 to centromeres. Compared to the targeting domain of other known and putative CenH3 species, Dictyostelium CenH3 has a shorter loop 1 region. The localisation of a variety of histone modifications and histone modifying enzymes was examined. Using fluorescence in situ hybridisation (FISH) and CenH3 chromatin-immunoprecipitation (ChIP) it was shown that the six telocentric centromeres contain all of the DIRS-1 and most of the DDT-A and skipper transposons. During interphase the centromeres remain attached to the centrosome resulting in a single CenH3 cluster which also contains the putative histone H3K9 methyltransferase SuvA, H3K9me3 and HP1 (heterochromatin protein 1). Except for the centromere cluster and a number of small foci at the nuclear periphery opposite the centromeres, the rest of the nucleus is largely devoid of transposons and heterochromatin associated histone modifications. At least some of the small foci correspond to the distal telomeres, suggesting that the chromosomes are organised in a Rabl-like manner. It was found that in contrast to metazoans, loading of CenH3 onto Dictyostelium centromeres occurs in late G2 phase. Transformation of Dictyostelium with vectors carrying the G418 resistance cassette typically results in the vector integrating into the genome in one or a few tandem arrays of approximately a hundred copies. In contrast, plasmids containing a Blasticidin resistance cassette integrate as single or a few copies. The behaviour of transgenes in the nucleus was examined by FISH, and it was found that low copy transgenes show apparently random distribution within the nucleus, while transgenes with more than approximately 10 copies cluster at or immediately adjacent to the centromeres in interphase cells regardless of the actual integration site along the chromosome. During mitosis the transgenes show centromere-like behaviour, and ChIP experiments show that transgenes contain the heterochromatin marker H3K9me2 and the centromeric histone variant H3v1. This clustering, and centromere-like behaviour was not observed on extrachromosomal transgenes, nor on a line where the transgene had integrated into the extrachromosomal rDNA palindrome. This suggests that it is the repetitive nature of the transgenes that causes the centromere-like behaviour. A Dictyostelium homolog of DET1, a protein largely restricted to multicellular eukaryotes where it has a role in developmental regulation was identified. As in other species Dictyostelium DET1 is nuclear localised. In ChIP experiments DET1 was found to bind the promoters of a number of developmentally regulated loci. In contrast to other species where it is an essential protein, loss of DET1 is not lethal in Dictyostelium, although viability is greatly reduced. Loss of DET1 results in delayed and abnormal development with enlarged aggregation territories. Mutant slugs displayed apparent cell type patterning with a bias towards pre-stalk cell types.

Adaptive Real-time Anomaly-based Intrusion Detection using Data Mining and Machine Learning Techniques

Die zunehmende Vernetzung der Informations- und Kommunikationssysteme führt zu einer weiteren Erhöhung der Komplexität und damit auch zu einer weiteren Zunahme von Sicherheitslücken. Klassische Schutzmechanismen wie Firewall-Systeme und Anti-Malware-Lösungen bieten schon lange keinen Schutz mehr vor Eindringversuchen in IT-Infrastrukturen. Als ein sehr wirkungsvolles Instrument zum Schutz gegenüber Cyber-Attacken haben sich hierbei die Intrusion Detection Systeme (IDS) etabliert. Solche Systeme sammeln und analysieren Informationen von Netzwerkkomponenten und Rechnern, um ungewöhnliches Verhalten und Sicherheitsverletzungen automatisiert festzustellen. Während signatur-basierte Ansätze nur bereits bekannte Angriffsmuster detektieren können, sind anomalie-basierte IDS auch in der Lage, neue bisher unbekannte Angriffe (Zero-Day-Attacks) frühzeitig zu erkennen. Das Kernproblem von Intrusion Detection Systeme besteht jedoch in der optimalen Verarbeitung der gewaltigen Netzdaten und der Entwicklung eines in Echtzeit arbeitenden adaptiven Erkennungsmodells. Um diese Herausforderungen lösen zu können, stellt diese Dissertation ein Framework bereit, das aus zwei Hauptteilen besteht. Der erste Teil, OptiFilter genannt, verwendet ein dynamisches "Queuing Concept", um die zahlreich anfallenden Netzdaten weiter zu verarbeiten, baut fortlaufend Netzverbindungen auf, und exportiert strukturierte Input-Daten für das IDS. Den zweiten Teil stellt ein adaptiver Klassifikator dar, der ein Klassifikator-Modell basierend auf "Enhanced Growing Hierarchical Self Organizing Map" (EGHSOM), ein Modell für Netzwerk Normalzustand (NNB) und ein "Update Model" umfasst. In dem OptiFilter werden Tcpdump und SNMP traps benutzt, um die Netzwerkpakete und Hostereignisse fortlaufend zu aggregieren. Diese aggregierten Netzwerkpackete und Hostereignisse werden weiter analysiert und in Verbindungsvektoren umgewandelt. Zur Verbesserung der Erkennungsrate des adaptiven Klassifikators wird das künstliche neuronale Netz GHSOM intensiv untersucht und wesentlich weiterentwickelt. In dieser Dissertation werden unterschiedliche Ansätze vorgeschlagen und diskutiert. So wird eine classification-confidence margin threshold definiert, um die unbekannten bösartigen Verbindungen aufzudecken, die Stabilität der Wachstumstopologie durch neuartige Ansätze für die Initialisierung der Gewichtvektoren und durch die Stärkung der Winner Neuronen erhöht, und ein selbst-adaptives Verfahren eingeführt, um das Modell ständig aktualisieren zu können. Darüber hinaus besteht die Hauptaufgabe des NNB-Modells in der weiteren Untersuchung der erkannten unbekannten Verbindungen von der EGHSOM und der Überprüfung, ob sie normal sind. Jedoch, ändern sich die Netzverkehrsdaten wegen des Concept drif Phänomens ständig, was in Echtzeit zur Erzeugung nicht stationärer Netzdaten führt. Dieses Phänomen wird von dem Update-Modell besser kontrolliert. Das EGHSOM-Modell kann die neuen Anomalien effektiv erkennen und das NNB-Model passt die Änderungen in Netzdaten optimal an. Bei den experimentellen Untersuchungen hat das Framework erfolgversprechende Ergebnisse gezeigt. Im ersten Experiment wurde das Framework in Offline-Betriebsmodus evaluiert. Der OptiFilter wurde mit offline-, synthetischen- und realistischen Daten ausgewertet. Der adaptive Klassifikator wurde mit dem 10-Fold Cross Validation Verfahren evaluiert, um dessen Genauigkeit abzuschätzen. Im zweiten Experiment wurde das Framework auf einer 1 bis 10 GB Netzwerkstrecke installiert und im Online-Betriebsmodus in Echtzeit ausgewertet. Der OptiFilter hat erfolgreich die gewaltige Menge von Netzdaten in die strukturierten Verbindungsvektoren umgewandelt und der adaptive Klassifikator hat sie präzise klassifiziert. Die Vergleichsstudie zwischen dem entwickelten Framework und anderen bekannten IDS-Ansätzen zeigt, dass der vorgeschlagene IDSFramework alle anderen Ansätze übertrifft. Dies lässt sich auf folgende Kernpunkte zurückführen: Bearbeitung der gesammelten Netzdaten, Erreichung der besten Performanz (wie die Gesamtgenauigkeit), Detektieren unbekannter Verbindungen und Entwicklung des in Echtzeit arbeitenden Erkennungsmodells von Eindringversuchen.