T Cell Priming by Activated Nlrc5-Deficient Dendritic Cells Is Unaffected despite Partially Reduced MHC Class I Levels.

NLRC5, a member of the NOD-like receptor (NLR) protein family, has recently been characterized as the master transcriptional regulator of MHCI molecules in lymphocytes, in which it is highly expressed. However, its role in activated dendritic cells (DCs), which are instrumental to initiate T cell responses, remained elusive. We show in this study that, following stimulation of DCs with inflammatory stimuli, not only did NLRC5 level increase, but also its importance in directing MHCI transcription. Despite markedly reduced mRNA and intracellular H2-K levels, we unexpectedly observed nearly normal H2-K surface display in Nlrc5(-/-) DCs. Importantly, this discrepancy between a strong intracellular and a mild surface defect in H2-K levels was observed also in DCs with H2-K transcription defects independent of Nlrc5. Hence, alongside with demonstrating the importance of NLRC5 in MHCI transcription in activated DCs, we uncover a general mechanism counteracting low MHCI surface expression. In agreement with the decreased amount of neosynthesized MHCI, Nlrc5(-/-) DCs exhibited a defective capacity to display endogenous Ags. However, neither T cell priming by endogenous Ags nor cross-priming ability was substantially affected in activated Nlrc5(-/-) DCs. Altogether, these data show that Nlrc5 deficiency, despite significantly affecting MHCI transcription and Ag display, is not sufficient to hinder T cell activation, underlining the robustness of the T cell priming process by activated DCs.

Direct Interrogation of Viral Peptides Presented by the Class I HLA of HIV-Infected T Cells

Identification of CD8+ cytotoxic T lymphocyte (CTL) epitopes has traditionally relied upon testing of overlapping peptide libraries for their reactivity with T cells in vitro. Here, we pursued deep ligand sequencing (DLS) as an alternative method of directly identifying those ligands that are epitopes presented to CTLs by the class I human leukocyte antigens (HLA) of infected cells. Soluble class I HLA-A*11:01 (sHLA) was gathered from HIV-1 NL4-3-infected human CD4+ SUP-T1 cells. HLA-A*11:01 harvested from infected cells was immunoaffinity purified and acid boiled to release heavy and light chains from peptide ligands that were then recovered by size-exclusion filtration. The ligands were first fractionated by high-pH high-pressure liquid chromatography and then subjected to separation by nano-liquid chromatography (nano-LC)–mass spectrometry (MS) at low pH. Approximately 10 million ions were selected for sequencing by tandem mass spectrometry (MS/MS). HLA-A*11:01 ligand sequences were determined with PEAKS software and confirmed by comparison to spectra generated from synthetic peptides. DLS identified 42 viral ligands presented by HLA-A*11:01, and 37 of these were previously undetected. These data demonstrate that (i) HIV-1 Gag and Nef are extensively sampled, (ii) ligand length variants are prevalent, particularly within Gag and Nef hot spots where ligand sequences overlap, (iii) noncanonical ligands are T cell reactive, and (iv) HIV-1 ligands are derived from de novo synthesis rather than endocytic sampling. Next-generation immunotherapies must factor these nascent HIV-1 ligand length variants and the finding that CTL-reactive epitopes may be absent during infection of CD4+ T cells into strategies designed to enhance T cell immunity.

SMRT-mediated co-shuttling enables export of class IIa HDACs independent of their CaM kinase phosphorylation sites

The Class IIa histone deacetylases (HDAC)4 and HDAC5 play a role in neuronal survival and behavioral adaptation in the CNS. Phosphorylation at 2/3 N-terminal sites promote their nuclear export. We investigated whether non-canonical signaling routes to Class IIa HDAC export exist because of their association with the co-repressor Silencing Mediator Of Retinoic And Thyroid Hormone Receptors (SMRT). We found that, while HDAC5 and HDAC4 mutants lacking their N-terminal phosphorylation sites (HDAC4(MUT), HDAC5(MUT)) are constitutively nuclear, co-expression with SMRT renders them exportable by signals that trigger SMRT export, such as synaptic activity, HDAC inhibition, and Brain Derived Neurotrophic Factor (BDNF) signaling. We found that SMRT's repression domain 3 (RD3) is critical for co-shuttling of HDAC5(MUT), consistent with the role for this domain in Class IIa HDAC association. In the context of BDNF signaling, we found that HDAC5(WT), which was more cytoplasmic than HDAC5(MUT), accumulated in the nucleus after BDNF treatment. However, co-expression of SMRT blocked BDNF-induced HDAC5(WT) import in a RD3-dependent manner. In effect, SMRT-mediated HDAC5(WT) export was opposing the BDNF-induced HDAC5 nuclear accumulation observed in SMRT's absence. Thus, SMRT's presence may render Class IIa HDACs exportable by a wider range of signals than those which simply

Semantic domain and grammatical class effects in the picture-word interference paradigm

The role of grammatical class in lexical access and representation is still not well understood. Grammatical effects obtained in picture-word interference experiments have been argued to show the operation of grammatical constraints during lexicalization when syntactic integration is required by the task. Alternative views hold that the ostensibly grammatical effects actually derive from the coincidence of semantic and grammatical differences between lexical candidates. We present three picture-word interference experiments conducted in Spanish. In the first two, the semantic relatedness (related or unrelated) and the grammatical class (nouns or verbs) of the target and the distracter were manipulated in an infinitive form action naming task in order to disentangle their contributions to verb lexical access. In the third experiment, a possible confound between grammatical class and semantic domain (objects or actions) was eliminated by using action-nouns as distracters. A condition in which participants were asked to name the action pictures using an inflected form of the verb was also included to explore whether the need of syntactic integration modulated the appearance of grammatical effects. Whereas action-words (nouns or verbs), but not object-nouns, produced longer reaction times irrespective of their grammatical class in the infinitive condition, only verbs slowed latencies in the inflected form condition. Our results suggest that speech production relies on the exclusion of candidate responses that do not fulfil task-pertinent criteria like membership in the appropriate semantic domain or grammatical class. Taken together, these findings are explained by a response-exclusion account of speech output. This and alternative hypotheses are discussed.

Synthesis of angiotensin-converting enzyme (ACE) inhibitors: an important class of antihypertensive drugs

This report outlines the discovery, the design and development of new compounds, and, structure-activity relationships for this drug category. Updated approaches to planned syntheses of new worthy ACE-inhibitors are also exploited.

Synthesis, characterization, and biological activity of a new class of dialkylphosphorylhydrazone derivatives of isatin

Sixteen dialkylphosphorylhydrazones were synthesized by condensation of phosphorylhydrazines with substituted isatins. Products were characterized by FTIR, ¹H-NMR, 13C-NMR, and 31P-NMR. Fungicidal activities of these compounds against Rhizoctonia solani and Fusarium oxysporum were also evaluated. Some compounds inhibited the growth of Rhizoctonia solani and Fusarium oxysporum by 43% and 51%, respectively. These compounds exhibited no effects on germination of lettuce seeds (Lactuca sativa L).

Clinical evaluation of enalapril maleate and furosemide usage in dogs with degenerative myxomatous mitral valve, CHF functional class Ib

Degenerative myxomatous mitral valve (DMMV) is a heart disease of high incidence in small animal clinical medicine, affecting mainly older dogs and small breeds. Thus, a scientific investigation was performed in order to evaluate the clinical use of the medicines furosemide and enalapril maleate in dogs with this disease in CHF functional class Ib before and after the treatment was established. For this purpose 16 dogs with the given valve disease were used, separated into two groups: the first received furosemide (n=8) and the second received enalapril maleate (n=8) throughout 56 days. The dogs were evaluated in four stages (T0, T14, T28 and T56 day) in relation to clinical signs, hematological, biochemical and serum assessment, which included serum angiotensin converting enzyme (ACE) and aldosterone, as well as radiography, electrocardiography, Doppler-echocardiography and blood pressure. The results regarding the clinical, hematological and serum chemistry evaluations revealed no significant changes in both groups, but significant reductions in the values of ACE and aldosterone in the group receiving enalapril maleate were verified. The radiographic examination revealed reductions of VHS values and variable Pms wave of the electrocardiogram in both groups, but no changes in blood pressure values were identified. The echocardiogram showed a significant decrease of the variables LVDd/s in the studied groups and the FS% in animals that received only enalapril. Therefore, analysis of results showed that monotherapy based on enalapril maleate showed better efficiency of symptoms control in patients with CHF functional class Ib.

Sivistyksestä sivilisaatioon. Kulttuurikäsitys J. V. Snellmanin historiallisessa ajattelussa

From Bildung to Civilisation. Conception of Culture in J. V. Snellman’s Historical Thinking The research explores Johan Vilhelm Snellman’s (1806–1881) conception of culture in the context of his historical thinking. Snellman was a Finnish, Swedish-speaking journalist, teacher and thinker, who held a central position in the Finnish national discourse during the nineteenth century. He has been considered as one of the leading theorists of a Finnish nation, writing widely about the themes such as the advancement of the national education, Finnish language and culture. Snellman is already a widely studied person in Finnish intellectual history, often characterised as a follower of G. W. F. Hegel’s philosophical system. My own research introduces a new kind of approach on Snellman’s texts, emphasising the conceptual level of his thought. With this approach, my aim is to broaden the Finnish research tradition on conceptual history. I consider my study as a cultural history of concepts, belonging also to the field of intellectual history. My focus is on one hand on the close reading of Snellman’s texts and on the other hand on contextualising his texts to the European intellectual tradition of the time. A key concept of Snellman’s theoretical thinking is his concept of bildning, which can be considered as a Swedish counterpart of the German concept of Bildung. The Swedish word incorporated all the main elements of the German concept. It could mean education or the so-called high culture, but most fundamentally it was about the self-formation of the individual. This is also the context in which Snellman’s concept of bildning has often been interpreted. In the study, I use the concept of bildning as a starting point of my research but I broaden my focus on the cognate concepts such as culture (kultur), spirit (anda) and civilisation. The purpose of my study is thus to illustrate how Snellman used and modified these concepts and from these observations to draw a conclusion about the nature of his conception of culture. Snellman was an early Finnish philosopher of history but also interested in the practice of the writing of history. He did not write any historical presentations himself but followed the publications in the field of history and introduced European historical writing to the Finnish, Swedish-speaking reading audience in his newspapers. The primary source material consists of different types of Snellman’s texts, including philosophical writings, lecture material, newspaper articles and private letters. I’m reading Snellman’s texts in the context of other texts produced both by his Finnish predecessors and contemporaries and by Swedish, German and French writers. Snellman’s principal philosophical works, Versuch einer spekulativen Entwicklung der Idee der Persönlichkeit (1841) and Läran om staten (1842), were both written abroad. Both of the works were contributions to contemporary debates on the international level, especially in Germany and Sweden. During the 1840s and 1850s Snellman had two newspapers of his own, Saima and Litteraturblad, which were directed towards the Swedish-speaking educated class. Both of the newspapers were very popular and their circulations were among the largest of their day in Finland. The topics of his articles and reviews covered literature, poetry, philosophy and education as well as issues concerning the economic, industrial and technical development in Finland. In his newspapers Snellman not only brought forth his own ideas but also spread the knowledge of European events and ideas to his readers. He followed very carefully the cultural and political situation in Western Europe. He also followed European magazines and newspapers and was well acquainted with German, French and also English literature – and of course Swedish literature to with which he had the closest ties. In his newspapers Snellman wrote countless number of literary reviews and critics, introducing his readers to European literature. The study consists of three main chapters in which I explore my research question in three different, yet overlapping contexts. In the first of these chapters, I analyse Snellman’n theoretical thinking and his concepts of bildning, kultur, anda and civilisation in the context of earlier cultural discourse in Finland as well as the tradition of German idealistic philosophy and neo-humanism. With the Finnish cultural discourse I refer to the early cultural discussion in Finland, which emerged after the year 1809, when Finland became an autonomous entity of its own as a Grand Duchy of Russia. Scholars of the Academy of Turku opened a discussion on the themes such as the state of national consciousness, the need for national education and the development of the Finnish language as a national language of Finland. Many of these academics were also Snellman’s teachers in the early years of his academic career and Snellman clearly formulated his own ideas in the footsteps of these Finnish predecessors. In his theoretical thinking Snellman was a collectivist; according to him an individual should always be understood in connection with the society, its values and manners, as well as to the traditions of a culture where an individual belongs to. In his philosophy of the human spirit Snellman was in many ways a Hegelian but his notion of education or ‘bildning’ includes also elements that connect him with the wider tradition of German intellectual history, namely the neo-humanist tradition and, at least to some extent, to the terminology of J. G. Herder or J. G. Fichte, for example. In this chapter, I also explore Snellman’s theory of history. In his historical thinking Snellman was an idealist, believing in the historical development of the human spirit (Geist in German language). One can characterise his theory of history by stating that it is a mixture of a Hegelian triumph of the spirit and Herderian emphasis on humanity (Humanität) and the relative nature of ‘Bildung’. For Snellman, the process of ‘bildning’ or ‘Bildung’ is being realised in historical development through the actions of human beings. Snellman believed in the historical development of the human civilization. Still Snellman himself considered that he had abandoned Hegel’s idea about the process of world history. Snellman – rightly or wrongly – criticised Hegel of emphasising the universal end of history (the realisation of the freedom of spirit) at the expense of the historical plurality and the freedom of each historical era. Snellman accused Hegel of neglecting the value and independency of different historical cultures and periods by imposing the abstract norm, the fulfilment of the freedom of spirit, as the ultimate goal of history. The historicist in Snellman believed in the individuality of each historical period; each historical era or culture had values, traditions and modes of thought of its own. This historicist in Snellman could not accept the talk about one measure or the end of history. On the other hand Snellman was also a universalist. He believed that mankind had a common task and that task was the development of ‘Bildung’, freedom or humanity. The second main chapter consists of two parts. In the first part, I explore the Finnish nationalistic discourse from the cultural point of view by analysing the notions such as a nation, national spirit or national language and showing how Snellman formulated his own ideas in a dialogic situation, participating in the Finnish discourse but also reacting to international discussions on the themes of the nation and nationality. For Snellman nationality was to a great extent the collective knowledge and customs or practices of the nation. Snellman stated that nationality is to be considered as a form of ‘bildning’. This could be seen not simply as affection for the fatherland but also for the mental identity of the nation, its ways of thinking, its practices, national language, customs and laws, the history of the nation. The simplest definition of nationality that Snellman gives is that nationality is the social life of the people. In the second part of the chapter I exam Snellman’s historical thinking and his understanding about historical development, interaction between different nations and cultures in the course of history, as well as the question of historical change; how do cultures or civilisations develop and who are the creators of culture? Snellman did not believe in one dominating culture but understood the course of history as a dialogue between different cultures. On the other hand, his views are very Eurocentric – here he follows the ideas of Hegel or for example the French historian François Guizot – for Snellman Europe represented the virtue of pluralism; in Europe one could see the diversity of cultures which, on the other hand, were fundamentally based on a common Christian tradition. In the third main chapter, my focus is on the writing of history, more precisely on Snellman’s ideas on the nature of history as a science and on the proper way of writing historical presentations. Snellman wrote critics on the works of history and introduced his readers to the writing of history especially in France, Sweden and German-speaking area – in some extend also in Britain. Snellman’s collectivistic view becomes evident also in his reviews on historical writing. For Snellman history was not about the actions of the states and their heads, nor about the records of ruling families and battles fought. He repeatedly stressed that history is a discipline that seeks to provide a total view of a phenomenon. A historian should not only collect information on historical events, since this information touches only the surface of a certain epoch or civilisation; he has to understand an epoch as totality. This required an understanding about the major contours in history, connections between civilisations and an awareness of significant turning points in historical development. In addition, it required a holistic understanding about a certain culture or historical era, including also the so-called inner life of a specific nation, a common people and their ways of life. Snellman wrote explicitly about ‘cultural history’ in his texts, referring to this kind of broad understanding of a society. In historical writing Snellman found this kind of broader view from the works of the French historians such as François Guizot and Jules Michelet. In all of these chapters, I elaborate the conceptual dimension of Snellman’s historical thinking. In my study I argue that Snellman not only adopted the German concepts of Bildung or Kultur in his own thinking but also developed the Swedish concepts in a way that include personal and innovative aspects. Snellman’s concept of bildning is not only a translation from ‘Bildung’ but he uses the Swedish concept in a versatile way that includes both the moral aspect of human development and social dimension of a human life. Along with ‘bildning’ Snellman used also the terms ‘kultur’ and ‘civilisation’ when referring to the totality of a certain nation or historical era, including both the so-called high culture (arts, science, religion) and the modes of thought as well as ways of life of the people as a whole. Unlike many of his Finnish contemporaries, Snellman did not use civilisation as a negative concept, lacking the moral essence of German term ‘Bildung’ or ‘Kultur’. Instead, for Snellman civilisation was a neutral term and here he comes close to the French tradition of using the term. In the study I argue that Snellman’s conception of culture in fact includes a synthesis of the German tradition of ‘Bildung’ and the French tradition of ‘civilisation’.

Local Features in Image and Video Processing - Object Class Matching and Video Shot Detection

The usage of digital content, such as video clips and images, has increased dramatically during the last decade. Local image features have been applied increasingly in various image and video retrieval applications. This thesis evaluates local features and applies them to image and video processing tasks. The results of the study show that 1) the performance of different local feature detector and descriptor methods vary significantly in object class matching, 2) local features can be applied in image alignment with superior results against the state-of-the-art, 3) the local feature based shot boundary detection method produces promising results, and 4) the local feature based hierarchical video summarization method shows promising new new research direction. In conclusion, this thesis presents the local features as a powerful tool in many applications and the imminent future work should concentrate on improving the quality of the local features.

The improvement of enclosure protection class of power electronics device

Tässä diplomityössä tutkittiin vaihtoehtoja tehoelektroniikkalaitteiden kotelointiluokan kehittämiseksi. Haasteena paremman suojauksen suunnittelussa on laitteiden tuottama suuri määrä lämpöä, joka vaatii tehokkaan jäähdytyksen. Työn tuloksena saatu prototyyppi IP33 luokkaa varten täyttää standardissa SFS-EN 60529+A1 asetetut vaatimukset kyseiselle kotelointiluokalle. Rakenteessa ja valmistettavuudessa havaittiin muutama ongelma, jotka ovat korjattavissa pienillä muutoksilla. Korkeampia suojausluokkia varten testattiin IP54-luokiteltujen filtterituulettimien vaikutusta laitteen jäähdytykseen. Testien perusteella jäähdytysteho on riittävä ja filtterituulettimet todettiin toimivaksi ratkaisuksi korkeammille suojausluokille. Työn perusteella voidaan todeta, että nykyiset laitteet voidaan muokata vastaamaan IP33 luokan vaatimuksia kohtuullisen pienillä muutoksilla. Tätä korkeammat suojausluokat vaatisivat niin suuria muutoksia designiin, että todennäköisesti täysin uuden laitteen suunnittelu olis kannattavin vaihtoehto.

Regulation of T cell response to leishmania antigens by determinants of histocompatibility leukocyte class I and II molecules

It has been shown that HLA class I molecules play a significant role in the regulation of the proliferation of T cells activated by mitogens and antigens. We evaluated the ability of mAb to a framework determinant of HLA class I molecules to regulate T cell proliferation and interferon gamma (IFN-g) production against leishmania, PPD, C. albicans and tetanus toxoid antigens in patients with tegumentary leishmaniasis and healthy subjects. The anti-major histocompatibility complex (MHC) mAb (W6/32) suppressed lymphocyte proliferation by 90% in cultures stimulated with aCD3, but the suppression was variable in cultures stimulated with leishmania antigen. This suppression ranged from 30-67% and was observed only in 5 of 11 patients. IFN-g production against leishmania antigen was also suppressed by anti-HLA class I mAb. In 3 patients IFN-g levels were suppressed by more than 60%, while in the other 2 cultures IFN-g levels were 36 and 10% lower than controls. The suppression by HLA class I mAb to the proliferative response in leishmaniasis patients and in healthy controls varied with the antigens and the patients or donors tested. To determine whether the suppression is directed at antigen presenting cells (APCs) or at the responding T cells, experiments with antigen-primed non-adherent cells, separately incubated with W6/32, were performed. Suppression of proliferation was only observed when the W6/32 mAb was added in the presence of T cells. These data provide evidence that a mAb directed at HLA class I framework determinants can suppress proliferation and cytokine secretion in response to several antigens.

Sleep-wake pattern of medical students: early versus late class starting time

The sleep-wake cycle of students is characterized by delayed onset, partial sleep deprivation and poor sleep quality. Like other circadian rhythms, the sleep-wake cycle is influenced by endogenous and environmental factors. The aim of the present study was to determine the effects of different class starting times on the sleep-wake pattern of 27 medical students. The data were collected during two medical school semesters having different class starting times. All subjects answered the Portuguese version of the Horne and Östberg Morningness/Eveningness Questionnaire, the Pittsburgh Sleep Quality Index (PSQI) and kept a sleep diary for two weeks during each semester. Better sleep quality (PSQI = 5.3 vs 3.4), delayed sleep onset (23:59 vs 0:54 h) and longer sleep duration (6 h and 55 min vs 7 h and 25 min) were observed with the late schedule. We also found reduced sleep durations during weekdays and extended sleep durations during weekends. This pattern was more pronounced during the semester with the early class schedule, indicating that the students were more sleep deprived when their classes began earlier in the morning. These results require further investigation regarding the temporal organization of our institutions.

Viral membrane fusion: is glycoprotein G of rhabdoviruses a representative of a new class of viral fusion proteins?

Enveloped viruses always gain entry into the cytoplasm by fusion of their lipid envelope with a cell membrane. Some enveloped viruses fuse directly with the host cell plasma membrane after virus binding to the cell receptor. Other enveloped viruses enter the cells by the endocytic pathway, and fusion depends on the acidification of the endosomal compartment. In both cases, virus-induced membrane fusion is triggered by conformational changes in viral envelope glycoproteins. Two different classes of viral fusion proteins have been described on the basis of their molecular architecture. Several structural data permitted the elucidation of the mechanisms of membrane fusion mediated by class I and class II fusion proteins. In this article, we review a number of results obtained by our laboratory and by others that suggest that the mechanisms involved in rhabdovirus fusion are different from those used by the two well-studied classes of viral glycoproteins. We focus our discussion on the electrostatic nature of virus binding and interaction with membranes, especially through phosphatidylserine, and on the reversibility of the conformational changes of the rhabdovirus glycoprotein involved in fusion. Taken together, these data suggest the existence of a third class of fusion proteins and support the idea that new insights should emerge from studies of membrane fusion mediated by the G protein of rhabdoviruses. In particular, the elucidation of the three-dimensional structure of the G protein or even of the fusion peptide at different pH's might provide valuable information for understanding the fusion mechanism of this new class of fusion proteins.