Epidemiological and functional characterization of Streptococcus pneumoniae pili

Streptococcus pneumoniae is an important life threatening human pathogen causing agent of invasive diseases such as otitis media, pneumonia, sepsis and meningitis, but is also a common inhabitant of the respiratory tract of children and healthy adults. Likewise most streptococci, S. pneumoniae decorates its surface with adhesive pili, composed of covalently linked subunits and involved in the attachment to epithelial cells and virulence. The pneumococcal pili are encoded by two genomic regions, pilus islet 1 (PI-1), and pilus islet-2 (PI-2), which are present in about 30% and 16% of the pneumococcal strains, respectively. PI-1 exists in three clonally related variants, whereas PI-2 is highly conserved. The presence of the islets does not correlate with the serotype of the strains, but with the genotype (as determined by Multi Locus Sequence Typing). The prevalence of PI-1 and PI-2 positive strains is similar in isolates from invasive disease and carriage. To better dissect a possible association between PIs presence and disease we evaluated the distribution of the two PIs in a panel of 113 acute otitis media (AOM) clinical isolates from Israel. PI-1 was present in 30.1% (N=34) of the isolates tested, and PI-2 in 7% (N=8). We found that 50% of the PI-1 positive isolates belonged to the international clones Spain9V-3 (ST156) and Taiwan19F-14 (ST236), and that PI-2 was not present in the absence of Pl-1. In conclusion, there was no correlation between PIs presence and AOM, and, in general, the observed differences in PIs prevalence are strictly dependent upon regional differences in the distribution of the clones. Finally, in the AOM collection the prevalence of PI-1 was higher among antibiotic resistant isolates, confirming previous indications obtained by the in silico analysis of the MLST database collection. Since the pilus-1 subunits were shown to confer protection in mouse models of infection both in active and passive immunization studies, and were regarded as potential candidates for a new generation of protein-based vaccines, the functional characterization was mainly focused on S. pneumoniae pilus -1 components. The pneumococcal pilus-1 is composed of three subunits, RrgA, RrgB and RrgC, each stabilized by intra-molecular isopeptide bonds and covalently polymerized by means of inter-molecular isopeptide bonds to form an extended fibre. The pilus shaft is a multimeric structure mainly composed by the RrgB backbone subunit. The minor ancillary proteins are located at the tip and at the base of the pilus, where they have been proposed to act as the major adhesin (RrgA) and as the pilus anchor (RrgC), respectively. RrgA is protective in in vivo mouse models, and exists in two variants (clades I and II). Mapping of the sequence variability onto the RrgA structure predicted from X-ray data showed that the diversity was restricted to the “head” of the protein, which contains the putative binding domains, whereas the elongated “stalk” was mostly conserved. To investigate whether this variability could influence the adhesive capacity of RrgA and to map the regions important for binding, two full-length protein variants and three recombinant RrgA portions were tested for adhesion to lung epithelial cells and to purified extracellular matrix (ECM) components. The two RrgA variants displayed similar binding abilities, whereas none of the recombinant fragments adhered at levels comparable to those of the full-length protein, suggesting that proper folding and structural arrangement are crucial to retain protein functionality. Furthermore, the two RrgA variants were shown to be cross-reactive in vitro and cross-protective in vivo in a murine model of passive immunization. Taken together, these data indicate that the region implicated in adhesion and the functional epitopes responsible for the protective ability of RrgA may be conserved and that the considerable level of variation found within the “head” domain of RrgA may have been generated by immunologic pressure without impairing the functional integrity of the pilus.

L'attitudine alla comunicazione. Uno studio nel Corso di Laurea in Infermieristica

It is believed that the way of being and the communicative-relational skills of every individual have multifactorial origins, including the quality of primary relationships with caregivers. For some time, the need for health care professionals to possess specific communicative and interpersonal skills has been highlighted. To the degree course in Nursing, like to all other degree programs related to health, access is granted to students who have large individual differences, both in terms of personality, and in terms of relational skills. Each academic year, therefore, the people responsible for the didactic organization of every course, are faced with having to prepare a training plan capable of addressing communicative-relational aspects and, at the same time, of being adequate to the real attitudes of incoming students. Thus, the need for appropriate tools for measuring the personological and vocational traits considered specific to health professions was born. This study has a twofold objective. On one hand, it aims at selecting a battery of psychological tests to detect psychological and attitudinal patterns, to facilitate the coordinators of graduate courses in their didactic organization and planning of educational training; on the other hand, it seeks to assess the correlations between communicative-relational skills (Relational-Communicative style, according to the model of patient-centered medicine-TRS) (Mucchielli’s Test of Spontaneous Attitudes – usual kind of attitude in dual relationships), personality traits (Alexithymia), styles of attachment to parental figures (PBI), and the capability of recognizing facial emotions, in a sample of students enrolled in the first year of a degree in Nursing.

Design, synthesis and application of ultrastable rylene dyes for fluorescent labeling of biomolecules

Studies of organic fluorescent dyes are experiencing a renaissance related to the increasing demands posed by new microscopy techniques for high resolution and high sensitivity. While in the last decade single molecule equipment and methodology has significantly advanced and in some cases reached theoretical limits (e.g. detectors approaching unity quantum yields) unstable emission from chromophores and photobleaching become more and more the bottleneck of the advancement and spreading of single-molecule fluorescence studies. The main goal of this work was the synthesis of fluorophores that are water-soluble, highly fluorescent in an aqueous environment, have a reactive group for attachment to a biomolecule and posses exceptional photostability. An approach towards highly fluorescent, water-soluble and monofunctional perylene-3,4,9,10-tetracarboxdiimide and terrylene-3,4:11,12-tetra carboxidiimide chromophores was presented. A new synthetic strategy for the desymmetrization of perylenetetracarboximides was elaborated; water-solubility was accomplished by introducing sulfonyl substituents in the phenoxy ring. Two strategies have been followed relying on either non-specific or site specific labeling. For this purpose a series of new water-soluble monofunctional perylene and terrylene dyes, bearing amine or carboxy group were prepared. The reactivity and photophysical properties of these new chromophores were studied in aqueous medium. The most suitable chromophores were further derivatized with amine or thiol reactive groups, suitable for chemical modification of proteins. The performance of the new fluorescent probes was assessed by single molecule enzyme tracking, in this case phospholipase acting on phospholipid supported layers. Phospholipase-1 (PLA-1) was labeled with N-hydroxysuccinimide ester functionalized perylene and terrylene derivatives. The purification of the conjugates was accomplished by novel convenient procedure for the removal of unreacted dye from labeled enzymes, which involves capturing excess dye with a solid support. This novel strategy for purification of bioconjugates allows convenient and fast separation of labeled proteins without the need for performing time consuming chromatographic or electrophoretic purification steps. The outstanding photostability of the dyes and, associated therewith, the extended survival times under strong illumination conditions allow a complete characterization of enzyme action on its natural substrates and even connecting enzyme mobility to catalytic activity. For site-specific attachment of the rylene dyes to proteins the chromophores were functionalized with thioesters or nitrilotriacetic acid groups. This allowed attachment of the emitters to the N-terminus of proteins by native chemical ligation or complexation with His-tagged polypeptides at the N- or C-termini, respectively. The synthesis of a water-soluble perylenebis (dicarboximide) functionalized with a thioester group was presented. This chromophore exhibits an exceptional photostability and a functional unit for site-specific labeling of proteins. The suitability of the fluorophore as a covalent label was demonstrated via native chemical ligation with protein containing N-terminal cystein residue. We exploited also oligohisitidine sequences as recognition elements for site-selective labeling. The synthesis of a new water-soluble perylene chromophore, containing a nitrilotriacetic acid functional group was demonstrated, using solution-phase and solid-phase approaches. This chromophore combines the exceptional photophysical properties of the rylene dyes and a recognition unit for site-specific labeling of proteins. An important feature of the label is the unchanged emission of the dye upon complexation with nickel ions.

Patient-Therapeut-Bindung bei Drogenabhängigen: Instrumentenvalidierung und Therapieevaluation

In der vorliegenden Arbeit wird zum einen ein Instrument zur Erfassung der Patient-Therapeut-Bindung validiert (Client Attachment to Therapist Scale, CATS; Mallinckrodt, Coble & Gantt, 1995), zum anderen werden Hypothesen zu den Zusammenhängen zwischen Selbstwirksamkeitserwartung, allgemeinem Bindungsstil, therapeutischer Beziehung (bzw. Therapiezufriedenheit), Patient-Therapeut-Bindung und Therapieerfolg bei Drogen-abhängigen in stationärer Postakutbehandlung überprüft. In die Instrumentenvalidierung (einwöchiger Retest) wurden 119 Patienten aus 2 Kliniken und 13 Experten einbezogen. Die Gütekriterien des Instrumentes fallen sehr zufriedenstellend aus. An der naturalistischen Therapieevaluationsstudie (Prä-, Prozess-, Post-Messung: T0, T1, T2) nahmen 365 Patienten und 27 Therapeuten aus 4 Kliniken teil. Insgesamt beendeten 44,1% der Patienten ihren stationären Aufenthalt planmäßig. Auf Patientenseite erweisen sich Alter und Hauptdiagnose, auf Therapeutenseite die praktizierte Therapierichtung als Therapieerfolgsprädiktoren. Selbstwirksamkeitserwartung, allgemeiner Bindungsstil, Patient-Therapeut-Bindung und Therapiezufriedenheit eignen sich nicht zur Prognose des Therapieerfolgs. Die zu T0 stark unterdurchschnittlich ausgeprägte Selbstwirksamkeits-erwartung steigert sich über den Interventionszeitraum, wobei sich ein Moderatoreffekt der Patient-Therapeut-Bindung beobachten lässt. Es liegt eine hohe Prävalenz unsicherer allgemeiner Bindungsstile vor, welche sich über den Therapiezeitraum nicht verändern. Die patientenseitige Zufriedenheit mit der Therapie steigt von T1 zu T2 an. Die Interrater-Konkordanz (Patient/Therapeut) zur Einschätzung der Patient-Therapeut-Bindung erhöht sich leicht von T1 zu T2. Im Gegensatz dazu wird die Therapiezufriedenheit von Patienten und Therapeuten zu beiden Messzeitpunkten sehr unterschiedlich beurteilt. Die guten Testgütekriterien der CATS sprechen für eine Überlegenheit dieses Instrumentes gegenüber der Skala zur Erfassung der Therapiezufriedenheit. Deshalb sollte die Patient-Therapeut-Bindung anhand dieses Instrumentes in weiteren Forschungsarbeiten an anderen Patientenkollektiven untersucht werden, um generalisierbare Aussagen zur Validität treffen zu können.

Valutazione affettiva del setting odontoiatrico in età evolutiva

Lo studio si pone l'obiettivo di approfondire il tema della valutazione affettiva del setting odontoiatrico in età evolutiva indagando se possa essere modulata da fattori quali: differenze di genere, specifici quadri patologici (Displasia Ectodermica e Patologie Sistemiche Croniche) responsabili di pregresse esperienze odontoiatriche e/o ospedalizzazioni; stato affettivo del bambino, stato affettivo del genitore. Materiali e metodi Studio 1. 85 soggetti [39 (19 maschi e 20 femmine) affetti da PSC e 46 (26 maschi e 20 femmine) soggetti sani] (range di età: 5-14 anni). Sono stati somministrati: un compito di valutazione immagini con 36 immagini (12 a contenuto piacevole, 12 neutro e 12 spiacevole) dallo IAPS e 12 immagini a contenuto odontoiatrico e dei questionari (MCDASf, CFSS-DS, TAD). Studio 2. 45 soggetti (19 maschi affetti da EDs e 26 maschi sani) (range di età: 5-14 anni). Sono stati somministrati: il compito di valutazione immagini precedentemente descritto e i questionari (MCDASf, CFSS-DS, TAD). Studio 3. 104 bambini (64 maschi and 40 femmine) (range di età: 5-14 anni) e uno dei genitori (19 padri and 69 madri). Sono stati somministrati i questionari MCDASf, CFSS-DS, TAD ai bambini e FDPQ, STAI-Y1, Y2-BDI-II ai genitori. Risultati: il contesto odontoiatrico ha una caratterizzazione affettiva distinta rispetto a contesti piacevoli, spiacevoli o neutri; specifici quadri patologici (EDs e PSC) non sembrano modulare il tipo di valutazione affettiva del contesto odontoiatrico; le femmine, attribuiscono al contesto odontoiatrico una valenza significativamente più spiacevole e una maggiore attivazione emotiva rispetto ai maschi; la paura del dolore odontoiatrico del genitore ha una correlazione con l’ansia dei bambini. Conclusione: è importante che l’odontoiatra consideri che la risposta emotiva delle bambine può essere caratterizzata da vissuti di paura con tendenza a comportamenti di allontanamento e valuti se la figura genitoriale rappresenta una risorsa o un elemento disturbante nella corso della seduta stessa.

Yoruba nationalist movements, ethnic politics and violence : a creation from historical consciousness and socio-political space in South-western Nigeria /// Arbeitspapiere / Institut für Ethnologie und Afrikastudien ; 105

Since 1900, the Yoruba people of South-western Nigeria have put its ethnic history at work in the construction of its identity in Nigeria. The exercise resulted in the creation of ethno-nationalist movements and the practice of ethnic politics, often expressed through violent attacks on the Nigerian State and some ethnic groups in Nigeria. Relying on mythological attachment to its traditions and subjective creation of cultural pride, the people created a sense of history that established a common interest among different Yoruba sub-groups in form of pan-Yoruba interest which forms the basis for the people’s imagination of nation. Through this, historical consciousness and socio-political space in which Yoruba people are located acted as instrumental forces employed by Yoruba political elites, both at colonial and post-colonial periods to demand for increasing access to political and economic resources in Nigeria. In form of nationalism, nationalist movements and ethnic politics continued in South-western Nigeria since 1900, yet without resulting to actual creation of an independent Yoruba State up to 2009. Through ethnographic data, the part played by history, tradition and modernity is examined in this paper. While it is concluded that ethno-nationalist movement and ethnic politics in Yoruba society are constructive agenda dated back to pre-colonial period, it continues to transform both in structure and function. Thus, Yoruba ethno-nationalist movement and ethnic politics is ambiguous, dynamic and complex, to the extent that it remains a challenge to State actions in Nigeria.

La ville picaresque

Cet extrait (le début du projet) peut être essentiellement considéré comme un travail de synthèse, de traduction et d’argumentation concernant un des textes de référence parmi les plus importants au sujet de la littérature picaresque : “La literatura picaresca desde la historia social” écrit par Jose Antonio Maravall. Mon but en développant ce projet était de me focaliser sur les idées principales contenues dans ce volume et en particulier sur la relation existante entre la littérature picaresque et le concept de ville, en termes urbanistiques. Je tiens à souligner le fait que ce travail a été longuement approfondi dans mes nombreuses activités d’étude et de recherche et en me référant, volontairement, au texte lui-même. La raison fondamentale de mon désir de rester près du texte se justifie par le fait que je venais de commence ce qui seraient mes futures activités de recherche de doctorat. J’ai utilisé ces idées à propos de la littérature picaresque, idées qui sont aujourd’hui universellement acceptées, afin de m’orienter moi-même dans ce genre littéraire. Les autres idées que j’ai étudiées et décrites au cours de mes activités de recherche successives sont plus personnelles, car j’avais eu l’occasion de les analyser auparavant. Ce travail n’inclut pas seulement des activités de recherche , mais il s’est agi aussi d’un projet de traduction puisque j’ai dû traduire de l’espagnol(texte original) en italien, pour ensuite concevoir et puis rédiger ma synthèse et mon argumentation en langue française. En conclusion, je peux affirmer que j’ai été capable de construire de solides bases pour élaborer et planifier ma thèse de Doctorat, avec les typiques thèmes de la littérature picaresque, pour exemple la recherche de la nourriture dans la ville.

Studio longitudinale sulle capacità cognitive del cane: discriminazione di quantità ed apprendimento per imitazione in relazione all'attaccamento

Il presente studio ha indagato e valutato alcune abilità cognitive del cane: la capacità di discriminare quantità e le capacità di apprendimento mediante imitazione; quest’ultima è poi stata messa in relazione con l’attaccamento nei confronti del proprietario. Per l’esecuzione della prima indagine sono stati messi appunto due test: il primo si è basato esclusivamente sulla presentazione di uno stimolo visivo: diversi quantitativi di cibo, differenti tra loro del 50%, sono stati presentati al cane; la scelta effettuata dai soggetti testati è stata premiata con differenti tipi di rinforzo differenziale o non differenziale. Il secondo test è stato diviso in due parti: sono stati presentati al cane diversi quantitativi di cibo sempre differenti tra loro del 50% ma nella prima parte del test l’input sensoriale per il cane è stato esclusivamente uditivo mentre nella seconda parte è stato sia uditivo che visivo. Ove è stato possibile è stato applicato ai cani un cardiofrequenzimetro al fine di eseguire una valutazione delle variazioni della frequenza cardiaca nel corso del test. Lo scopo è stato quello di valutare se i soggetti testati erano in grado di discriminare la quantità maggiore. La seconda indagine ha analizzato le capacità di apprendimento di 36 soggetti che sono stati suddivisi in cani da lavoro e pet. I soggetti protagonisti dello studio hanno eseguito il Mirror Test per la valutazione dell’apprendimento per imitazione. I soggetti presi in considerazione, sono stati sottoposti a scansione termografica all’inizio ed al termine del test ed è stata rilevata la loro frequenza respiratoria nella fase iniziale e finale del test. In 11 soggetti che hanno eseguito il precedente test è stato possibile eseguire anche il Strange Situation Test per la valutazione dell’attaccamento al proprietario; i test in questione sono stati videoregistrati ed analizzati per mezzo di un software preposto (OBSERVER XT 10).

Lipid remodelling of glycosylphosphatidylinositol (GPI) glycoconjugates in procyclic-form trypanosomes: biosynthesis and processing of GPIs revisited

The African trypanosome, Trypanosoma brucei, has been used as a model to study the biosynthesis of GPI (glycosylphosphatidylinositol) anchors. In mammalian (bloodstream)-form parasites, diacyl-type GPI precursors are remodelled in their lipid moieties before attachment to variant surface glycoproteins. In contrast, the GPI precursors of insect (procyclic)-form parasites, consisting of lyso-(acyl)PI (inositol-acylated acyl-lyso-phosphatidylinositol) species, remain unaltered before protein attachment. By using a combination of metabolic labelling, cell-free assays and complementary MS analyses, we show in the present study that GPI-anchored glycoconjugates in T. congolense procyclic forms initially receive tri-acylated GPI precursors, which are subsequently de-acylated either at the glycerol backbone or on the inositol ring. Chemical and enzymatic treatments of [3H]myristate-labelled lipids in combination with ESI-MS/MS (electrospray ionization-tandem MS) and MALDI-QIT-TOF-MS3 (matrix-assisted laser-desorption ionization-quadrupole ion trap-time-of-flight MS) analyses indicate that the structure of the lipid moieties of steady-state GPI lipids from T. congolense procyclic forms consist of a mixture of lyso-(acyl)PI, diacyl-PI and diacyl-(acyl)PI species. Interestingly, some of these species are myristoylated at the sn-2 position. To our knowledge, this is the first demonstration of lipid remodelling at the level of protein- or polysaccharide-linked GPI anchors in procyclic-form trypanosomes.

Eukaryotic translation elongation factor 1A (eEF1A) domain I from S. cerevisiae is required but not sufficient for inter-species complementation

Ethanolamine phosphoglycerol (EPG) is a protein modification attached exclusively to eukaryotic elongation factor 1A (eEF1A). In mammals and plants, EPG is linked to conserved glutamate residues located in eEF1A domains II and III, whereas in the unicellular eukaryote Trypanosoma brucei, only domain III is modified by a single EPG. A biosynthetic precursor of EPG and structural requirements for EPG attachment to T. brucei eEF1A have been reported, but nothing is known about the EPG modifying enzyme(s). By expressing human eEF1A in T. brucei, we now show that EPG attachment to eEF1A is evolutionarily conserved between T. brucei and Homo sapiens. In contrast, S. cerevisiae eEF1A, which has been shown to lack EPG is not modified in T. brucei. Furthermore, we show that eEF1A cannot functionally complement across species when using T. brucei and S. cerevisiae as model organisms. However, functional complementation in yeast can be obtained using eEF1A chimera containing domains II or III from other species. In contrast, yeast domain I is strictly required for functional complementation in S. cerevisiae.

The Sociolinguistics of Hungarian Outside Hungary

This study describes the sociolinguistic situation of the indigenous Hungarian national minorities in Slovakia (c. 600,000), Ukraine (c. 180,000), Romania (c. 2,000,000), Yugoslavia (c. 300,000), Slovenia (c. 8,000) and Austria (c. 6,000). Following the guidelines of Hans Goebl et al, the historical sociolinguistic portrait of each minority is presented from 1920 through to the mid-1990s. Each country's report includes sections on geography and demography, history, politics, economy, culture and religion, language policy and planning, and language use (domains of minority and/or majority language use, proficiency, attitudes, etc.). The team's findings were presented in the form of 374 pages of manuscripts, articles and tables, written in Hungarian and English. The core of the team's research results lies in the results of an empirical survey designed to study the social characteristics of Hungarian-minority bilingualism in the six project countries, and the linguistic similarities and differences between the six contact varieties of Hungarian and Hungarian in Hungary. The respondents were divided by age, education, and settlement group - city vs. village and local majority vs. local minority. The first thing to be observed is that Hungarian is tending to be spoken less to children than to parents and grandparents, a familiar pattern of language shift. In contact varieties of Hungarian, analytic constructions may be used where monolingual Hungarians would use a more synthetic form. Mr Kontra gives as an example the compound tagdij, which in Standard Hungarian means "membership fee" but which is replaced in contact Hungarian by the two-word phrase tagsagi dij. Another similar example concerns the synthetic verb hegedult "played the violin" and the analytic expression hegedun jatszott. The contrast is especially striking between the Hungarians in the northern Slavic countries, who use the synthetic form frequently, and those in the southern Slavic countries, who mainly use the analytic form. Mr. Kontra notes that from a structural point of view, there is no immediate explanation for this, since Slovak or Ukrainian are as likely to cause interference as is Serbian. He postulates instead that the difference may be attributable to some sociohistoric cause, and points out that the Turkish occupation of what is today Voivodina caused a discontinuity of the Hungarian presence in the region, with the result that Hungarians were resettled in the area only two and a half centuries ago. However, the Hungarians in today's Slovakia and Ukraine have lived together with Slavic peoples continuously for over a millennium. It may be, he suggests, that 250 years of interethnic coexistence is less than is needed for such a contact-induced change to run its course. Next Mr. Kontra moved on to what he terms "mental maps and morphology". In Hungarian, the names of cities and villages take the surface case (eg. Budapest-en "in Budapest") whereas some names denoting Hungarian settlements and all names of foreign cities take the interior case (eg. Tihany-ban "in Tihany" and Boston-ban "in Boston). The role of the semantic feature "foreign" in suffix-choice can be illustrated by such minimal pairs as Velence-n "in Velence, a village in Hungary" versus Velence-ben "in Velence [=Venice], a city in Italy", and Pecs-en "in Pecs, a city in Hungary" vs. Becs-ben "in Becs, ie. Vienna". This Hungarian vs. foreign distinction is often interpreted as "belonging to historical (pre-1920) Hungary" vs. "outside historical Hungary". The distinction is also expressed in the dichotomy "home" vs. "abroad'. The 1920 border changes have had an impact on both majority and minority Hungarians' mental maps, the maps which govern the choice of surface vs. interior cases with placenames. As there is a growing divergence between the mental maps of majority and minority Hungarians, so there will be a growing divergence in their use of the placename suffixes. Two placenames were chosen to scratch the surface of this complex problem: Craiova (a city in Oltenia, Romania) and Kosovo (Hungarian Koszovo) an autonomous region in southeast Yugoslavia. The assumption to be tested was that both placenames would be used with the inessive (interior) suffixes categorically by Hungarians in Hungary, but that the superessive suffix (showing "home") would be used near-categorically by Hungarians in Romania and Yugoslavia (Voivodina). Minority Hungarians in countries other than Romania and Yugoslavia would show no difference from majority Hungarians in Hungary. In fact, the data show that, contrary to expectation, there is considerable variation within Hungary. And although Koszovo is used, as expected, with the "home" suffix by 61% of the informants in Yugoslavia, the same suffix is used by an even higher percentage of the subjects in Slovenia. Mr. Kontra's team suggests that one factor playing a role in this might be the continuance of the former Yugoslav mentality in the Hungarians of Slovenia, at least from the geographical point of view. The contact varieties of Hungarian show important grammatical differences from Hungarian in Hungary. One of these concerns the variable use of Null subjects (the inclusion or exclusion of the subject of the verb). When informants were asked to insert either megkertem or megkertem ot - "I asked her" - into a test sentence, 54.9% of the respondents in the Ukraine inserted the second phrase as opposed to only 27.4% in Hungary. Although Mr. Kontra and his team concentrated more on the differences between Contact Hungarian and Standard Hungarian, they also discovered a number of similarities. One such similarity is demonstrable in the distribution of what Mr. Kontra calls an ongoing syntactic merger in Hungarian in Hungary. This change means effectively that two possibilities merge to form a third. For instance, the two sentences Valoszinuleg kulfoldre fognak koltozni and Valoszinu, hogy kulfoldre fognak koltozni merge to form the new construction Valszinuleg, hogy kulfoldre fognak koltozni ("Probably they will move abroad."). When asked to choose "the most natural" of the sentences, one in four chose the new construction, and a chi-square test shows homogeneity in the sample. In other words, this syntactic change is spreading across the entire Hungarian-speaking region in the Carpathian Basin Mr. Kontra believes that politicians, educators, and other interested parties now have reliable and up-to-date information about each Hungarian minority. An awareness of Hungarian as a pluricentric language is being developed which elevates the status of contact varieties of Hungarian used by the minorities, an essential process, he believes, if minority languages are to be maintained.

DOCK2 is required for chemokine-promoted human T lymphocyte adhesion under shear stress mediated by the integrin alpha4beta1

The alpha4beta1 integrin is an essential adhesion molecule for recruitment of circulating lymphocytes into lymphoid organs and peripheral sites of inflammation. Chemokines stimulate alpha4beta1 adhesive activity allowing lymphocyte arrest on endothelium and subsequent diapedesis. Activation of the GTPase Rac by the guanine-nucleotide exchange factor Vav1 promoted by CXCL12 controls T lymphocyte adhesion mediated by alpha4beta1. In this study, we investigated the role of DOCK2, a lymphocyte guanine-nucleotide exchange factor also involved in Rac activation, in CXCL12-stimulated human T lymphocyte adhesion mediated by alpha4beta1. Using T cells transfected with DOCK2 mutant forms defective in Rac activation or with DOCK2 small interfering RNA, we demonstrate that DOCK2 is needed for efficient chemokine-stimulated lymphocyte attachment to VCAM-1 under shear stress. Flow chamber, soluble binding, and cell spreading assays identified the strengthening of alpha4beta1-VCAM-1 interaction, involving high affinity alpha4beta1 conformations, as the adhesion step mainly controlled by DOCK2 activity. The comparison of DOCK2 and Vav1 involvement in CXCL12-promoted Rac activation and alpha4beta1-dependent human T cell adhesion indicated a more prominent role of Vav1 than DOCK2. These results suggest that DOCK2-mediated signaling regulates chemokine-stimulated human T lymphocyte alpha4beta1 adhesive activity, and that cooperation with Vav1 might be required to induce sufficient Rac activation for efficient adhesion. In contrast, flow chamber experiments using lymph node and spleen T cells from DOCK2(-/-) mice revealed no significant alterations in CXCL12-promoted adhesion mediated by alpha4beta1, indicating that DOCK2 activity is dispensable for triggering of this adhesion in mouse T cells, and suggesting that Rac activation plays minor roles in this process.

Barn owl (Tyto alba) siblings vocally negotiate resources

Current theory proposes that nestlings beg to signal hunger level to parents honestly, or that siblings compete by escalating begging to attract the attention of parents. Although begging is assumed to be directed at parents, barn owl (Tyto alba) nestlings vocalize in the presence but also in the absence of the parents. Applying the theory of asymmetrical contests we experimentally tested three predictions of the novel hypothesis that in the absence of the parents siblings vocally settle contests over prey items to be delivered next by a parent. This 'sibling negotiation hypothesis' proposes that offspring use each others begging vocalization as a source of information about their relative willingness to contest the next prey item delivered. In line with the hypothesis we found that (i) a nestling barn owl refrains from vocalization when a rival is more hungry, but (ii) escalates once the rival has been fed by a parent, and (iii) nestlings refrain from and escalate vocalization in experimentally enlarged and reduced broods, respectively. Thus, when parents are not at the nest a nestling vocally refrains when the value of the next delivered prey item will be higher for its nest-mates. These findings are the exact opposite of what current models predict for begging calls produced in the presence of the parents. [References: 20]

A parent-completed respiratory questionnaire for 1-year-old children: repeatability

BACKGROUND AND AIMS: There are few standardised questionnaires for the assessment of respiratory symptoms in preschool children. We have developed and tested the short-term repeatability of a postal questionnaire on respiratory symptoms for 1-year-old children. METHODS: A newly developed postal questionnaire for the assessment of wheeze and other respiratory symptoms was sent to parents of a population-based random sample of 4300 children aged 12-24 months. After an interval of 3 months, a random sample of 800 respondents received the questionnaire a second time. The responses were compared using Cohen's kappa (kappa) to assess agreement corrected for chance. RESULTS: The first questionnaire was returned by 3194 (74%) families, the second one by 460/800 (58%). Repeatability was excellent (kappa 0.80-0.96) for questions on household characteristics, environmental exposures and family history, good (kappa 0.61-0.80) for questions on prevalence, severity and treatment of wheeze, and moderate (kappa 0.39-0.66) for chronic cough and upper respiratory symptoms. CONCLUSIONS: This short postal questionnaire designed for use in population-based studies has excellent repeatability for family and household characteristics and good repeatability for questions on wheeze. Short-term changes in symptom status might be responsible for variable answers on recent chronic cough and upper respiratory symptoms. Overall, the questionnaire is a valuable instrument for community-based research on respiratory symptoms in 1 to 2-year-old children.

Morphogenesis of the peri-implant mucosa: an experimental study in dogs

PURPOSE: The objective of the present experiment was to study the morphogenesis of the mucosal attachment to implants made of c.p. titanium. MATERIAL AND METHODS: All mandibular premolars were extracted in 20 Labrador dogs. After a healing period of 3 months, four implants (ITI Dental Implant System) were placed in the right and left sides of the mandible. A non-submerged implant installation technique was used and the mucosal tissues were secured to the conical marginal portion of the implants with interrupted sutures. The sutures were removed after 2 weeks and a plaque control program including daily cleaning of the remaining teeth and the implants was initiated. The animals were sacrificed and biopsies were obtained at various intervals to provide healing periods extending from Day 0 (2 h) to 12 weeks. The mandibles were removed and placed in the fixative. The implant sites were dissected using a diamond saw and processed for histological analysis. RESULTS: Large numbers of neutrophils infiltrated and degraded the coagulum that occupied the compartment between the mucosa and the implant during the initial phase of healing. At 2 weeks after surgery, fibroblasts were the dominating cell population in the connective tissue interface but at 4 weeks the density of fibroblasts had decreased. Furthermore, the first signs of epithelial proliferation were observed in specimens representing 1-2 weeks of healing and a mature barrier epithelium occurred after 6-8 weeks of healing. The collagen fibers of the mucosa were organized after 4-6 weeks of healing. CONCLUSION: It is suggested that the soft-tissue attachment to implants placed using a non-submerged installation procedure is properly established after several weeks following surgery.