Auto Poisoning of the Respiratory Chain by a Quorum-Sensing-Regulated Molecule Favors Biofilm Formation and Antibiotic Tolerance.

Bacterial programmed cell death and quorum sensing are direct examples of prokaryote group behaviors, wherein cells coordinate their actions to function cooperatively like one organism for the benefit of the whole culture. We demonstrate here that 2-n-heptyl-4-hydroxyquinoline-N-oxide (HQNO), a Pseudomonas aeruginosa quorum-sensing-regulated low-molecular-weight excreted molecule, triggers autolysis by self-perturbing the electron transfer reactions of the cytochrome bc1 complex. HQNO induces specific self-poisoning by disrupting the flow of electrons through the respiratory chain at the cytochrome bc1 complex, causing a leak of reducing equivalents to O2 whereby electrons that would normally be passed to cytochrome c are donated directly to O2. The subsequent mass production of reactive oxygen species (ROS) reduces membrane potential and disrupts membrane integrity, causing bacterial cell autolysis and DNA release. DNA subsequently promotes biofilm formation and increases antibiotic tolerance to beta-lactams, suggesting that HQNO-dependent cell autolysis is advantageous to the bacterial populations. These data identify both a new programmed cell death system and a novel role for HQNO as a critical inducer of biofilm formation and antibiotic tolerance. This newly identified pathway suggests intriguing mechanistic similarities with the initial mitochondrial-mediated steps of eukaryotic apoptosis.

Ager Tarraconensis 5. Paisatge, poblament, cultura material i història = Landscape, Settlement, Material Culture and History Proceedings of the International Symposium

Aquesta obra recull 28 comunicacions presentades al Simposi internacional «L’ager Tarraconensis. Paisatge, poblament, cultura material i història», celebrat a Tarragona el 27 i 28 d’octubre del 2010, organitzat per l’Institut Català d’Arqueologia Clàssica a la seu de l’Institut i a la Diputació de Tarragona. Les aportacions al simposi se centren en l'evolució del paisatge al Camp de Tarragona i en altres regions del món romà, en la varietat de les primeres estratègies de control romà del territori, en les tipologies i evolució dels establiments, en aspectes econòmics i en la dinàmica del poblament, sense faltar-hi alguna aportació de l'epigrafia. Tot plegat fa referència molt especialment al territori de Tarraco, tot i que el simposi també va comptar amb comunicacions sobre altres territoris de l'àrea catalana i saguntina i d'altres províncies romanes, com la Lusitània, les Gàl·lies i la Transpadana; les comparacions entre diferents regions han donat una riquesa notable al debat. Entre els resultats més suggerents comptem amb la constatació de la similitud en els models de romanització del territori en les diferents àrees d'àmbit català i valencià, que contrasten significativament amb els models de territoris d'Andalusia, França i Portugal.

¿Crisis de la cultura? = Crisis of the culture?

Culture has several meanings: civilization, creation, knowledge, life… We analyze how each one suffers the so-called crisis of culture. Civilization lives the crisis as «liquidity», according to Bauman’s diagnosis, with significant negative impact on education. The creation and enjoyment of cultural products, such as museums, suffer its crisis as fragility resulting from the civilization crisis, notably in the form of consumerist banality or pure entertainment. Culture as knowledge and as life is analyzed under the joint notion «humanistic culture». This, which in turn is creation, knowledge and life has its specific corruption in the elitist knowledge, merely theoretical. The hallmark of genuine humanism contains an essential component, the dimension of ethical and political commitment. The crisis in the humanistic studies, noted by Nussbaum, threats the values of humanistic culture; in particular is a political risk, because democracy is a system that needs to sustain and improve the values of humanistic culture. In the background, and beyond the differences between the cultural meanings, there is a unique cultural crisis, a crisis of ethics and politics at a time

From "being to the self" to "become to be": on development of autonomy construction in nursing students

Objective: To understand nursing student's self-consciousness and his/her autonomy in the discipline of fundamentals of professional care in the context of a liberating pedagogical proposal. Methodology. This qualitative, case-based research in the model of Ludke and André involved 14 students participating in the discipline. Data were collected by non-participatory observation and analysis of documents. Field observation was conducted from March to July 2010 and data were collected according to the proposal of Minayo: pre-analysis, exploration of material and treatment of results. Results. We constructed two thematic units of analysis: from "being to the self" and exercise of "become to be". Conclusion. When nursing students feel more liberty, they have the opportunity to substitute the scary prospect of learning something new material to something that motivates their curiosity and leads them to become more autonomous.

Chemotaxonomy of three genera of the annonaceae family using self-organizing maps and 13C NMR data of diterpenes

The Annonaceae family is distributed throughout Neotropical regions of the world. In Brazil, it covers nearly all natural formations particularly Annona, Xylopia and Polyalthia and is characterized chemically by the production of sources of terpenoids (mainly diterpenes), alkaloids, steroids, polyphenols and, flavonoids. Studies from 13C NMR data of diterpenes related with their botanical occurrence were used to generate self-organizing maps (SOM). Results corroborate those in the literature obtained from morphological and molecular data for three genera and the model can be used to project other diterpenes. Therefore, the model produced can predict which genera are likely to contain a compound.

Methods for construction and analysis of computational models in systems biology : applications to the modelling of the heat shock response and the self-assembly of intermediate filaments

Systems biology is a new, emerging and rapidly developing, multidisciplinary research field that aims to study biochemical and biological systems from a holistic perspective, with the goal of providing a comprehensive, system- level understanding of cellular behaviour. In this way, it addresses one of the greatest challenges faced by contemporary biology, which is to compre- hend the function of complex biological systems. Systems biology combines various methods that originate from scientific disciplines such as molecu- lar biology, chemistry, engineering sciences, mathematics, computer science and systems theory. Systems biology, unlike “traditional” biology, focuses on high-level concepts such as: network, component, robustness, efficiency, control, regulation, hierarchical design, synchronization, concurrency, and many others. The very terminology of systems biology is “foreign” to “tra- ditional” biology, marks its drastic shift in the research paradigm and it indicates close linkage of systems biology to computer science. One of the basic tools utilized in systems biology is the mathematical modelling of life processes tightly linked to experimental practice. The stud- ies contained in this thesis revolve around a number of challenges commonly encountered in the computational modelling in systems biology. The re- search comprises of the development and application of a broad range of methods originating in the fields of computer science and mathematics for construction and analysis of computational models in systems biology. In particular, the performed research is setup in the context of two biolog- ical phenomena chosen as modelling case studies: 1) the eukaryotic heat shock response and 2) the in vitro self-assembly of intermediate filaments, one of the main constituents of the cytoskeleton. The range of presented approaches spans from heuristic, through numerical and statistical to ana- lytical methods applied in the effort to formally describe and analyse the two biological processes. We notice however, that although applied to cer- tain case studies, the presented methods are not limited to them and can be utilized in the analysis of other biological mechanisms as well as com- plex systems in general. The full range of developed and applied modelling techniques as well as model analysis methodologies constitutes a rich mod- elling framework. Moreover, the presentation of the developed methods, their application to the two case studies and the discussions concerning their potentials and limitations point to the difficulties and challenges one encounters in computational modelling of biological systems. The problems of model identifiability, model comparison, model refinement, model inte- gration and extension, choice of the proper modelling framework and level of abstraction, or the choice of the proper scope of the model run through this thesis.

Bordering with Culture(s): Europeanization and Cultural Agency at the External Border of the European Union.

Rajallistamisen kulttuuri(t): Eurooppalaistuminen ja kulttuurinen toimijuus Euroopan unionin ulkorajalla Euroopan integraation ja Euroopan unionin laajentumisen myötä EU:n sisärajat ovat avautuneet kun taas sen ulkorajoilla lisääntyvää rajan ylitysten valvontaa on pyritty kompensoimaan yhteistyön merkitystä ja verkostoitumista painottamalla. Tämä tutkimus pyrkii ymmärtämään EU:n ulkorajan muutosten merkitystä paikalliselle hyvinvoinnille sekä laajemmin ylirajaisuuden merkitystä identiteettien rakentamiselle raja-alueilla. EU:n ulkorajalla Puolassa ja Suomessa toteutettavat rajat ylittävät, kulttuuriin ja kulttuuriperintöön liittyvät, projektit kertovat eurooppalaistumisesta ja sen vaikutuksista kulttuurisen horisontin muutokselle. Voidaan nähdä miten Eurooppa kulttuurisena konstruktiona tulee paikallisesti merkittäväksi tavoilla jotka kertovat myös paikallisten toimijoiden mahdollisuuksista osallistua EU-rajan muutoksia ja paikallisuutta määritteleviin prosesseihin. Tällöin erityisesti raja-alueiden materiaalisen perinnön, ja sen mahdollistamien rajaan liittyvien neuvottelujen, voidaan nähdä kertovat Eurooppalaistumisesta myös ns. alhaaltapäin muotoutuvana prosessina. Artikkeliväitöskirjan taustalla on Puolan ja Suomen toisen maailmansodan seurauksena luovuttamien raja-alueiden (Kresy ja Karjala) asema nykyisellä Euroopan unionin ulkorajalla. Tutkimusidea perustuu tutkijan omiin kokemuksiin projekteista Puolan ja Ukrainan raja-alueella vuonna 2003, ennen Puolan liittymistä EU:hun vuonna 2004. Tutkimusaineisto on peräisin vuosien 2005-2009 aikana tehdyistä ns. monipaikkaisista (multi-sited) kenttätöistä EU:n ulkorajalla, pääosin Puolassa ja Suomessa, joissa kohteena olivat kulttuuria ja kulttuuriperintöä hyödyntävät, pääosin EU-rahoitetut, rajat ylittävät projektit. Materiaalit koostuvat 34 projektitoimijan haastatteluista, projektien materiaaleista, kenttätöiden havainnoista, paikallisten sanomalehtien artikkeleista sekä eri tasoilla (EU, kansallinen, alueellinen) tuotetuista ohjelmadokumenteista. Huomio kiinnittyy projektitoimijoiden tapoihin tehdä rajanylityksiä, sekä heidän tapaansa kokea ja hyödyntää raja-alueiden kulttuuriperintöä sekä ymmärtää niiden nykyistä kulttuurista moninaisuutta. Tällöin havaitaan miten erilaiset eurooppalaiset ideat, representaatiot ja käytänteet tulevat osaksi erilaisia translokaaleja, rajat ylittäviä ja paikallis-eurooppalaisia, suhteita. Vertailun kohteeksi eivät tällöin asetu projektit, toimijat tai raja-alueet sinänsä, vaan näihin suhteisiin liittyvä kulttuurinen toimijuus. Keskeinen käsite tutkimuksessa on ’rajallistaminen’, eli sen havaitseminen, miten jokainen rajan ylitys tarkoittaa myös neuvottelua rajasta. Rajan ylitys voi siis tarkoittaa myös sen vahvistamista. Myös itse raja voi asettua toiminnan kohteeksi, jolloin nousee esiin se, miten rajat ylittäviä ”kulttuureja” käytetään ja mitkä ovat niiden rajaan liittyvät paikalliset merkitykset. Kysymys on siitä kuka, ja kenelle, rajan merkityksiä neuvottelee? Projektitoimijoiden voidaan nähdä neuvottelevan näitä erilaisia ”kulttuureja” jotka tuottavat rajaa neuvottelevia suhteita, kuten esimerkiksi yhteistyön verkostojen tapaa ohittaa rajan paikallinen merkitys. Tämä rajallistaminen voi kuitenkin tarkoittaa myös paikallisten kulttuuristen identifikaatioiden huomioimista. Tällöin kyse on myös sen luovuuden havaitsemisesta, jota yksilöillä on kun he neuvottelevat näitä erilaisia rajallistamisen kulttuureja. Erityisesti toisen maailmansodan seurauksena valtiorajoista tuli vahvasti kansallisia kulttuureja erottavia, mutta nyt kulttuurisista rajoista neuvotellaan ja rajojen yli tapahtuva vuorovaikutus, sekä paikallisen ja Eurooppalaisen tason väliset suhteet, ja niiden moniäänisyys, nousevat tutkimuksen keskiöön. Tutkimuksen yhteenvedon kannalta keskeinen on kysymys raja/alueen kestävyydestä. Tyypillisesti verkostoitumista painottavan rajat ylittävän yhteistyön suhde paikalliseen yhteisöön voi jäädä häilyväksi. Tavoite paikallisen kulttuuriperinnön suojeluun ei itsessään vielä kerro sen merkityksestä paikalliselle hyvinvoinnille. Arvioinnin kannalta on hyödyllistä nähdä miten myös materiaalisella perinnöllä on toimijuutta osana paikallisuutta muokkaavia suhteita. Paikallisten asukkaiden kokemus rajasta voi edelleen olla että se ei ole muuttunut Neuvostoliiton ajoista, toisaalta EU:nkin voidaan toivoa määrittelemään rajansa vielä tarkemmin, jotta sen kansallinen luonne muuttuisi. Tutkimus nostaa esiin miten eurooppalaiset yhteistyötä ja kulttuurista moninaisuutta korostavat ideat ja käytänteet vaikuttavat erityisesti puolalaisten toimijoiden mahdollisuuksiin määritellä EU-rajaan liittyviä prosesseja osana paikallisia kulttuuriperinnön määrittelyjä. Paikallisten rajaan liittyvien kulttuuristen identifikaatioiden liittäminen osaksi projekteja ei kuitenkaan ole helppoa. Toisaalta rajan merkitys on sisäistetty osana arkea, toisaalta taas rajaan liittyvät suuret kertomukset kansallisena ja EU-rajana voivat olla etäännyttäviä tekijöitä. EU-raja, projektit ja monikulttuurisen perinnön autenttisuus ovat kuitenkin raja-alueen toimijoille ja yhteisöille mahdollisuus osallistua rajallistamiseen. Toiminnan kestävyyden kannalta kyse on pitkälti siitä avaako rajallistaminen paikallisen perinnön merkityksiä osana paikallis-eurooppalaisia suhteita.

Language of the Future, Language of the PRC –Representations of Putonghua in South China Morning Post

As the national language of the PRC, the world's growing economic power and the sovereign of Hong Kong, Putonghua is a language with multiple facets of relevance for the current Special Administrative Region. This paper seeks to explore and explain different representations of Putonghua in Hong Kong's leading English-language newspaper South China Morning Post in articles published between January 2012 and February 2013. The representations are studied in the context of the different discourses in which they appear, some of which feature language(s) as a central theme and some more marginally. An overview is first presented of the scholarly research on the most important developments in Hong Kong's complex language scene from the beginnings of the colony until present day, with the aim of detecting developments and attitudes with potential relevance or parallels to the context of Putonghua today. The paper then reflects on the media and its role in producing and perpetuating discourses in the society, before turning to more practical considerations on Hong Kong's English and Chinese language media and the role of South China Morning Post in it. The methods used in analysing the discourses are those of discourse analysis, with textual analysis as its starting point, in which close attention is paid to linguistic forms as the concrete representations of meanings in a text. Particularly the immediate contexts of the appearances of the word “Putonghua” in the articles were studied carefully to detect vocabulary, grammar and semantical choices as signs of different discourses, potentially also revealing fundamental underlying assumptions and other “hidden meanings” in the text. Some of the most distinctive discourses in which different representations of Putonghua appeared were the Instrumental value for the individual (in which Putonghua was represented as a form of social capital); Othering of the mainlanders (in which Putonghua served as a concrete marker of distinction); Belonging to China (Putonghua as a symbol of unity); and Cultural distinctiveness of Hong Kong (Putonghua as a threat to Hong Kong's history and culture, as embodied in Cantonese). Some of these discourses were more prominent than others, and for example the discourse of Belonging to China was relatively rarely enacted in Hongkongers' voices. In general, the findings were not surprising in the light of the history, but showed a fair degree of consistency with what has been written earlier about the languages and attitudes towards them in Hong Kong. It has often been noted that Putonghua and its relation with Cantonese is a matter linked with the social identity of the colony and its citizens. While it appeared that there were no strict taboos in the representations of Putonghua in the societal context, the possibility of self-censorship cannot be ruled out as a factor toning down political discourses in the representations.

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

Epithelium, a highly dynamic system, plays a key role in the homeostasis of the intestine. However, thus far a human intestinal epithelial cell line has not been established in many countries. Fetal tissue was selected to generate viable cell cultures for its sterile condition, effective generation, and differentiated character. The purpose of the present study was to culture human intestinal epithelial cells by a relatively simple method. Thermolysin was added to improve the yield of epithelial cells, while endothelin-3 was added to stimulate their growth. By adding endothelin-3, the achievement ratio (viable cell cultures/total cultures) was enhanced to 60% of a total of 10 cultures (initiated from 8 distinct fetal small intestines), allowing the generation of viable epithelial cell cultures. Western blot, real-time PCR and immunofluorescent staining showed that cytokeratins 8, 18 and mouse intestinal mucosa-1/39 had high expression levels in human intestinal epithelial cells. Differentiated markers such as sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV also showed high expression levels in human intestinal epithelial cells. Differentiated human intestinal epithelial cells, with the expression of surface markers (cytokeratins 8, 18 and mouse intestinal mucosa-1/39) and secretion of cytokines (sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV), may be cultured by the thermolysin and endothelin-3 method and maintained for at least 20 passages. This is relatively simple, requiring no sophisticated techniques or instruments, and may have a number of varied applications.

Co-culture of chondrocytes and bone marrow mesenchymal stem cells in vitro enhances the expression of cartilaginous extracellular matrix components

Chondrocytes and bone marrow mesenchymal stem cells (BMSCs) are frequently used as seed cells in cartilage tissue engineering. In the present study, we determined if the co-culture of rabbit articular chondrocytes and BMSCs in vitro promotes the expression of cartilaginous extracellular matrix and, if so, what is the optimal ratio of the two cell types. Cultures of rabbit articular chondrocytes and BMSCs were expanded in vitro and then cultured individually or at a chondrocyte:BMSC ratio of 4:1, 2:1, 1:1, 1:2, 1:4 for 21 days and cultured in DMEM/F12. BMSCs were cultured in chondrogenic induction medium. Quantitative real-time RT-PCR and Western blot were used to evaluate gene expression. In the co-cultures, type II collagen and aggrecan expression increased on days 14 and 21. At the mRNA level, the expression of type II collagen and aggrecan on day 21 was much higher in the 4:1, 2:1, and 1:1 groups than in either the articular chondrocyte group or the induced BMSC group, and the best ratio of co-culture groups seems to be 2:1. Also on day 21, the expression of type II collagen and aggrecan proteins in the 2:1 group was much higher than in all other groups. The results demonstrate that the co-culture of rabbit chondrocytes and rabbit BMSCs at defined ratios can promote the expression of cartilaginous extracellular matrix. The optimal cell ratio appears to be 2:1 (chondrocytes:BMSCs). This approach has potential applications in cartilage tissue engineering since it provides a protocol for maintaining and promoting seed-cell differentiation and function.

Culture of bovine ovarian follicle wall sections maintained the highly estrogenic profile under basal and chemically defined conditions

Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17β-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures.