Characterization of two M17 family members in Escherichia coli, Peptidase A and Peptidase B

Escherichia coil encodes two aminopeptidases belonging to the M17 family: Peptidase A (PepA) and Peptidase B (PepB). To gain insights into their substrate specificities, PepA or PepB were overexpressed in Delta pepN, which shows greatly reduced activity against the majority of amino acid substrates. Overexpression of PepA or PepB increases catalytic activity of several aminopeptidase substrates and partially rescues growth of Delta pepN during nutritional downshift and hightemperature stress. Purified PepA and PepB display broad substratespecificity and Leu, Lys, Met and Gly are preferred substrates. However, distinct differences are observed between these two paralogs: PepA is more stable at high temperature whereas PepB displays broader substrate specificity as it cleaves Asp and insulin B chain peptide. Importantly, this strategy, i.e. overexpression of peptidases in Delta pepN and screening a panel of substrates for cleavage, can be used to rapidly identify peptidases with novel substrate specificities encoded in genomes of different organisms. (C) 2010 Elsevier Inc. All rights reserved.

Evolution, Homology Conservation, and Identification of Unique Sequence Signatures in GH19 Family Chitinases

The discovery of GH (Glycoside Hydrolase) 19 chitinases in Streptomyces sp. raises the possibility of the presence of these proteins in other bacterial species, since they were initially thought to be confined to higher plants. The present study mainly concentrates on the phylogenetic distribution and homology conservation in GH19 family chitinases. Extensive database searches are performed to identify the presence of GH19 family chitinases in the three major super kingdoms of life. Multiple sequence alignment of all the identified GH19 chitinase family members resulted in the identification of globally conserved residues. We further identified conserved sequence motifs across the major sub groups within the family. Estimation of evolutionary distance between the various bacterial and plant chitinases are carried out to better understand the pattern of evolution. Our study also supports the horizontal gene transfer theory, which states that GH19 chitinase genes are transferred from higher plants to bacteria. Further, the present study sheds light on the phylogenetic distribution and identifies unique sequence signatures that define GH19 chitinase family of proteins. The identified motifs could be used as markers to delineate uncharacterized GH19 family chitinases. The estimation of evolutionary distance between chitinase identified in plants and bacteria shows that the flowering plants are more related to chitinase in actinobacteria than that of identified in purple bacteria. We propose a model to elucidate the natural history of GH19 family chitinases.

Kuviteltu aatelismies : Aateluus viholliskuvana ja itseymmärryksenä 1800-luvun Suomessa

An imagined nobleman Nobility as an enemy image and in-group identity in nineteenth-century Finland The focal point of this study is the difficult relationship between two seemingly very different 19th-century elite groups, the upwardly mobile bourgeois intelligentsia and the slowly declining traditional nobility. In the thinking of the bourgeois contender the two emerged as exact opposites, styled as conflicting ideal types: an outdated, exclusive, degenerate hereditary aristocracy versus a dynamic and progressive new force in society, recruited solely on the basis of personal merit, originating from the common people and representing the nation. The appearance of an important 19th-century novelty, print publicity, coincided with the emergence of the bourgeois intelligentsia. The institutions of the developing publishing industry were manned by the aspiring new group. The strengthening flow of progressive, democratic, nationalist ideas distributed via the printing presses carried an undercurrent of self-promotion. It transmitted to the developing readership the self-image of the new cultural bourgeoisie as the defender and benevolent educator of the nation. Having won the contest over the media, the intelligentsia was free to present its predecessor and rival as an enemy of the people. In its politics the nobility emerged as an ideal scapegoat, represented as the source for existing social evils, all if which would promptly go away after its disappearance. It also served as a black backcloth, against which the democratic, national, progressive bourgeois intelligentsia would shine more brightly. In order to shed light on the 19th-century process of (re)modelling the image of nobility as a public enemy I have used four different types of source materials. These include three genres of print publicity, ranging from popular historical and contemporary fiction to nonfictional presentations of national history and the news and political commentaries of the daily papers, complemented by another, originally oral type of publicity, the discussion protocols of the Finnish four-estate parliament. To counterpoint these I also analysed the public self-image of the nobility, particularly vis-à-vis the nationalist and democratic ethos of the modernising politics.

Identification of Specific DNA Binding Residues in the TCP Family of Transcription Factors in Arabidopsis

The TCP transcription factors control multiple developmental traits in diverse plant species. Members of this family share an similar to 60-residue-long TCP domain that binds to DNA. The TCP domain is predicted to form a basic helix-loop-helix ( bHLH) structure but shares little sequence similarity with canonical bHLH domain. This classifies the TCP domain as a novel class of DNA binding domain specific to the plant kingdom. Little is known about how the TCP domain interacts with its target DNA. We report biochemical characterization and DNA binding properties of a TCP member in Arabidopsis thaliana, TCP4. We have shown that the 58-residue domain of TCP4 is essential and sufficient for binding to DNA and possesses DNA binding parameters comparable to canonical bHLH proteins. Using a yeast-based random mutagenesis screen and site-directed mutants, we identified the residues important for DNA binding and dimer formation. Mutants defective in binding and dimerization failed to rescue the phenotype of an Arabidopsis line lacking the endogenous TCP4 activity. By combining structure prediction, functional characterization of the mutants, and molecular modeling, we suggest a possible DNA binding mechanism for this class of transcription factors.

Transfers of Family Businesses to Non-Family Buyers - The Selling Business Family Perspective

This study focuses on business families and how they handle transitions such as business transfers. It also tries to shift the balance of research away from successions and towards business transfers as a key topic for family business researchers. In addition, it contributes to the family business research field by further highlighting the importance of the various different contributions in the family business from business family members other than the entrepreneurial founder. Based on interviews with both business family members and business brokers, it appears as important for business families who are selling their family business that it is managed in a similar way in the future regardless of the shift in ownership and management. It is also important that the employees can stay with the business. However, employees are seldom regarded as potential buyers of the family business; most preferably, from the point of view of business family members, this should be somebody who is similar to themselves. Business transfers can be lengthy processes, but once the family business is sold, previous owners most often want to leave the family business. This disengagement can be difficult for business family members if they have not managed to build up some other identity outside the family business environment. Money may compensate for the loss in the short run, but something else is needed in the long run, since the management of money is usually not perceived as that interesting. A family business transfer can have great influence on the members of the business family who is selling, and therefore it is suggested that personal due diligence could be of some help when planning the transfer. That tool can help business family members to analyse their own personal situation, but it may also make it easier to understand how the other business family members feel about the forthcoming change. Everyone is influenced in different ways during a family business transfer, and awareness of this fact may make it easier for the whole business family to adjust to their new environment.

New family of thallium cuprate superconductors not containing calcium or barium: TlSrn+1−xLnxCunO2n+3+δ (Ln=La, Pr, or Nd)

The first two members of the new TlSrn+1−xLnxCunO2n+3+δ (Ln=La, Pr, or Nd) series of superconducting cuprates possessing 1021 and 1122 type structures are described. The n=1 (1021) members with Tcs around 40 K have electrons or holes as the majority charge carriers depending on x. The n=2 (1122) cuprate (Ln=Pr or Nd) shows a Tc in the 80–90 K range.

The Critical Role of N- and C-Terminal Contact in Protein Stability and Folding of a Family 10 Xylanase under Extreme Conditions

Background: Stabilization strategies adopted by proteins under extreme conditions are very complex and involve various kinds of interactions. Recent studies have shown that a large proportion of proteins have their N- and C-terminal elements in close contact and suggested they play a role in protein folding and stability. However, the biological significance of this contact remains elusive. Methodology: In the present study, we investigate the role of N- and C-terminal residue interaction using a family 10 xylanase (BSX) with a TIM-barrel structure that shows stability under high temperature,alkali pH, and protease and SDS treatment. Based on crystal structure,an aromatic cluster was identified that involves Phe4, Trp6 and Tyr343 holding the Nand C-terminus together; this is a unique and important feature of this protein that might be crucial for folding and stabilityunder poly-extreme conditions. Conclusion: A series of mutants was created to disrupt this aromatic cluster formation and study the loss of stability and function under given conditions. While the deletions of Phe4 resulted in loss of stability, removal of Trp6 and Tyr343 affected in vivo folding and activity. Alanine substitution with Phe4, Trp6 and Tyr343 drastically decreased stability under all parameters studied. Importantly,substitution of Phe4 with Trp increased stability in SDS treatment.Mass spectrometry results of limited proteolysis further demonstrated that the Arg344 residue is highly susceptible to trypsin digestion in sensitive mutants such as DF4, W6A and Y343A, suggesting again that disruption of the Phe4-Trp6-Tyr343 (F-W-Y) cluster destabilizes the N-and C-terminal interaction. Our results underscore the importance of N- and C-terminal contact through aromatic interactions in protein folding and stability under extreme conditions, and these results may be useful to improve the stability of other proteins under suboptimal conditions.

Bi2−xPbx(Ca, Sr)n+1CunO2n+4+δ (n = 1, 2, 3, and 4) family of superconductors

Superconducting oxides of the Bi1.5Pb0.5(Ca, Sr)n+1CunO2n+4+δ series with n = 1, 2, 3 and 4 have been characterized. The superconducting transition temperature increases markedly with n up to n = 3, but the Tc of the n = 4 member is not much higher than that of the n = 3 member. The Tc does not change significantly in Bi2−xPbxCaSr2Cu2O8+δ with x (0.1 < x ≤ 0.5).

Köyhyyden perintö : Tutkimus kulttuurisen tiedon sisällöistä ja jatkuvuuksista suomalaisissa elämäkerta- ja sananlaskuaineistoissa

The Legacy of Poverty. A Study of the substance and continuity of cultural knowledge in Finnish biographical and proverbial texts The study focuses on the idea of the cultural knowledge and shared understanding that ordinary people, folk , have of the concepts and ideas about rural based poverty in Finland. Throughout 19th century and well into 20th century, the majority of the population remained agrarian and poor. By the 1950s, most people still lived in rural areas and a majority of them earned their living primarily from agriculture and forestry. Urbanization proceeded rapidly from the 1960s onwards. Even though the Nordic welfare state was firmly established in Finland by the 1970s, old forms of agrarian poverty still remained in the culture. The source material for the study consists of 99 biographies and 502 proverbs. Biographical texts include written autobiographies and interviewed biographies. A primary analyzing concept is called a poverty speech. The poverty speech has been analyzed by providing answers to the following three questions: What connotations do people attach to poverty when they speak about it? What sort of social relations arise when people speak about poverty? How is the past experience of poverty constructed in the present and in the welfare state context? Cultural knowledge is a theoretical and analytical tool that enables people to categorize information. The three questions stated above are crucial in revealing the schematic structure that people use to communicate about agrarian poverty. Categories are analyzed and processed in terms of cultural themes that contain the ideals and stereotypes of spoken motif and sub-themes. The application of theoretical and analytical premises to the poverty speech has shown that there are four cultural themes. The first theme is Power. The social connotations in the poverty speech are mostly in relation to the better-off people. Poverty does not exist without an awareness of welfare, i.e. the understanding of a certain standard of welfare above that of one's own. The second theme is about family ties as a resource and welfare network. In poverty speech, marriage is represented as a means to upgrade one's livelihood. Family members are described as supporting one another, but at the same time as being antagonists. The third theme, Work represents the work ethic that is being connected to the poverty. Hard working as a representation is attached to eligibility for `a good life´ that in Finland was to become an owner-occupier of a cottage or a flat. The fourth theme is Security. The resentment of unfair treatment is expressed by using moral superiority and rational explanations. The ruling classes in the agrarian society are portrayed as being evil and selfish with no social conscience because they did not provide enough assistance to those who needed it. During the period when the welfare benefit system was undeveloped, the poor expected the wealthier people to make a contribution to the distribution of material wealth. In the premises of cultural knowledge, both oral and written traditions are about human thinking: they deal with topics, ideas and evaluations that are relevant to their bearers. Many elements expressed in poverty speech, such as classifications and customs derived from the rural world, have been carried over into the next generation in newer contexts and a different cultural environment. Keywords: cultural knowledge, cognitive categorization, poverty, life stories, proverbs

Suomen suuriruhtinaskunnan virkamiehistön yksityiskirjastot ja sensuurin kieltämä kirjallisuus niissä 1800-luvun alkuvuosikymmeninä

Tutkimus luotaa 1800-luvun alkuvuosikymmenten Suomen suuriruhtinaskunnan virkamiehistön poliittisia asenteita sensuuri-ja painovapauskysymysten valossa. Sadan virkamiehistöön kuuluneen henkilön yksityiskirjastot ja niiden sisältämät mahdolliset sensuurin kieltämät nimikkeet on seulottu Kansalliskirjaston kokoelmissa olevista 1800- luvun kirjahuutokauppaluetteloista vertailemalla niitä Kansallisarkistossa säilytettäviin sensuuriviranomaisten kiellettyjen teosten listoihin. Tutkimuksen keskeisimmiksi teemoiksi nousevat näin poliittis- sekä kirjahistorialliset, kirjallista kulttuuria koskettelevat ulottuvuudet. Virkamiehistön yksityiskirjastot mahdollisine kiellettyine nimikkeineen on esitelty tilastollisen taulukoinnin avulla kirjastojen koon, virkamiehistön hallintoalan, rankiluokan sekä alueellisen identiteetin mukaan jaoteltuina. Vaikka lähdeaineiston perusteella on tämänkaltaisen poliittisväritteisen kirjallisen kulttuurin tavoittaminen osaksi ongelmallista, ovat huutokauppaluettelot systemaattisesta ja säännöllisestä luonteestaan johtuen merkittävää ja satunnaisten inventaarioluetteloiden ohella lähestulkoon ainoaa käsillä olevaa yksityiskirjastoja koskevaa lähdeaineistoa. Suomen 1800-luvun valtiolliseen asemaan liittyviä historiantulkintoja on tutkimuksen edetessä pyritty kuljettamaan rinnakkain; niitä ovat toisaalta suomalaisten käsitykset maansa perustuslaillisesta asemasta ja toisaalta venäläisten viranomaisten tulkinnat Suomen asemasta yleisvaltakunnallisessa viitekehyksessä. Tämän ohella tutkimusaihetta on lähestytty sen ajalliseeen kontekstiin olennaisesti kytkeytyvien yhteiskunta-ja historianfilosofisten teorioiden avulla. Ne ovat olleet myös tutkimuksen kohteena olleille aikalaisille relevantti henkinen ulottuvuus. Tämä on perusteltua, koska filosofista ja valtiotieteellistä kirjallisuutta on ollut kirjastoissa paljon. Etiikasta ja moraalista lähtevä vapauden ongelma on ollut yksi tämänkaltainen yhteiskuntafilosofinen kysymys. Tutkimuksessa aika ajoin esiin nouseva aatteellinen vertailu muiden Euroopan maiden oloihin laajentaa tutkimuksen historiallisen viitekehyksen osaksi 1800-luvun eurooppalaisen henkisen aatemaailman maisemaa. Tutkimuksen varovaiset johtopäätökset juontuvat erilaisista näkökulmista. Kiellettyjä nimikkeitä löytyi hieman yli 30 % kaikista virkamiehistön yksityiskirjastoista, mutta kiellettyjen nimikkeiden määrät osoittautuivat kirjastokohtaisesti laskettuna pienehköiksi. Ilmiön voi näin väittää olleen toisaalta marginaalista, mutta toisaalta konkreettinen osoitus poliittiseen vakaumukseen viittaavasta asennoitumisesta; sensuuria esimerkiksi saatettiin tulkita perustuslaillisuutta ja Aleksanteri I:n antaman hallitsijanvakuutuksen taustaa vasten. Kielletyn kirjallisuuden hankkiminen sensuurilaeista piittaamatta saattoi myös olla kamppailua hyvänä pidetyn yhteiskuntajärjestyksen puolesta järjestelmän epäkohtia vastaan. Keisaria kohtaan tunnettiin lojaalisuutta, mutta hänen hallitsemaansa järjestelmää edustavia sensuuriviranomaisia kohtaan välttämättä ei. Tässä kohdin Suomen tilanteella oli vastaavuutta Saksan oloihin, j o s s a nuorhegeliläiset halusivat muuttaa yhteiskuntaa ideaalimpaan suuntaan. Venäjän keisarikuntaan kuuluminen toi mukanaan maallista hyvää, mutta henkisen kulttuurin puolella orientoiduttiin pääasiassa länteen, ja pitäydyttiin Ruotsin ajan luomassa perinnössä. Yksi keskeinen tästä kummunnut valistusaatteen traditionaalinen arvo oli itsensä vapaa kehittäminen, joten sensuurikysymyksissä ajauduttiin väistämättä törmäyskurssille valtiopoliittisen ja kulttuurisen identiteetin ristipaineessa. Vaikka valtiopoliittinen identiteetti vaihtui vuoden 1809 jälkeen, ei kulttuurisen identiteetin kohdalla näin käynyt. Tätä kautta vahvistuu kokonaiskuva 1700-luvun historian ja Ruotsin ajan perinnön tärkeydestä 1800-luvun henkiselle kehitykselle etenkin kirjallisen kulttuurin saralla.

Jac. Ahrenberg ja historian perintö : Restaurointisuunnitelmat Viipurin ja Turun linnoihin 1800-luvun lopussa

Jac. Ahrenberg and Patrimony Restoration Plans for Viipuri and Turku Castles at the End of the 19th Century This dissertation examines the unrealized restoration plans for two castles in the Grand Duchy of Finland one located at Viipuri (Vyborg, nowadays in Russia), the other at Turku (in Swedish, Åbo) during the last decades of the 19th century. Both castles were used as prisons, barracks and warehouses. From the middle of the 19th century on, their restoration and transformation into museums and "national monuments" were demanded in the newspapers. The prison reform in the 1860s stimulated the documentation and debate concerning their future, but it was only at the beginning of the 1880s when their restoration became an official state-run project. The undertaking was carried out by Johan Jacob (Jac.) Ahrenberg (1847 1914), architect of the National Board of Public Buildings. By combining written sources with drawings and photographs, this dissertation examines the restoration projects, the two castles' significance and the ways in which they were investigated by scholars. The plans are analyzed in connection with restoration practices in France and Sweden and in the context of contemporary discussions concerning national art and patrimony. The thesis argues that these former castles of the Swedish crown were used to manifest the western roots of Finnish law and order, the lineage of power and the capacity of the nation to defend itself. However, because of their symbolism, their restoration became a politically delicate question concerning the role of the Swedish heritage in Finland's nation-building process. According to Jac. Ahrenberg's plans, the two castles were to be restored to their assumed appearance at the time of the Vasa dynasty. Consequently, the structures would have resembled castles in Sweden. It is suggested that one aim of the restoration plans was to transform the two buildings into monuments testifying to the common history of Sweden and Finland. They were meant to consolidate the Swedish basis of Finnish culture and autonomy and thus to secure them against the threatening implications of Russian imperialism. It seems that along with the changing ideals of architectural restoration and the need for an original Finnish architectural heritage, the political connotations associated with the castles were one reason why Jac. Ahrenberg's restoration plans were never realized.