Embryonic gene expression of Coregonus palaea (whitefish) under pathogen stress as analyzed by high-throughput RNA-sequencing.

Most fishes produce free-living embryos that are exposed to environmental stressors immediately following fertilization, including pathogenic microorganisms. Initial immune protection of embryos involves the chorion, as a protective barrier, and maternally-allocated antimicrobial compounds. At later developmental stages, host-genetic effects influence susceptibility and tolerance, suggesting a direct interaction between embryo genes and pathogens. So far, only a few host genes could be identified that correlate with embryonic survival under pathogen stress in salmonids. Here, we utilized high-throughput RNA-sequencing in order to describe the transcriptional response of a non-model fish, the Alpine whitefish Coregonus palaea, to infection, both in terms of host genes that are likely manipulated by the pathogen, and those involved in an early putative immune response. Embryos were produced in vitro, raised individually, and exposed at the late-eyed stage to a virulent strain of the opportunistic fish pathogen Pseudomonas fluorescens. The pseudomonad increased embryonic mortality and affected gene expression substantially. For example, essential, upregulated metabolic pathways in embryos under pathogen stress included ion binding pathways, aminoacyl-tRNA-biosynthesis, and the production of arginine and proline, most probably mediated by the pathogen for its proliferation. Most prominently downregulated transcripts comprised the biosynthesis of unsaturated fatty acids, the citrate cycle, and various isoforms of b-cell transcription factors. These factors have been shown to play a significant role in host blood cell differentiation and renewal. With regard to specific immune functions, differentially expressed transcripts mapped to the complement cascade, MHC class I and II, TNF-alpha, and T-cell differentiation proteins. The results of this study reveal insights into how P. fluorescens impairs the development of whitefish embryos and set a foundation for future studies investigating host pathogen interactions in fish embryos.

Long-term persistence of immunity induced by OVA-coupled gas-filled microbubble vaccination partially protects mice against infection by OVA-expressing Listeria.

Vaccination aims at generating memory immune responses able to protect individuals against pathogenic challenges over long periods of time. Subunit vaccine formulations based on safe, but poorly immunogenic, antigenic entities must be combined with adjuvant molecules to make them efficient against infections. We have previously shown that gas-filled microbubbles (MB) are potent antigen-delivery systems. This study compares the ability of various ovalbumin-associated MB (OVA-MB) formulations to induce antigen-specific memory immune responses and evaluates long-term protection toward bacterial infections. When initially testing dendritic cells reactivity to MB constituents, palmitic acid exhibited the highest degree of activation. Subcutaneous immunization of naïve wild-type mice with the OVA-MB formulation comprising the highest palmitic acid content and devoid of PEG2000 was found to trigger the more pronounced Th1-type response, as reflected by robust IFN-γ and IL-2 production. Both T cell and antibody responses persisted for at least 6 months after immunization. At that time, systemic infection with OVA-expressing Listeria monocytgenes was performed. Partial protection of vaccinated mice was demonstrated by reduction of the bacterial load in both the spleen and liver. We conclude that antigen-bound MB exhibit promising properties as a vaccine candidate ensuring prolonged maintenance of protective immunity.

Emergence and development of a financial cluster: the evolution of Andorra’s banking deposits in the long-term, 1931-2007

This paper explores the origins of Andorra’s financial cluster. It shows that the free movement of currency, the protection of infant industry, and geographical concentration lie at the foundation of the cluster’s competitive advantage. Drawing on a new set of data, the paper also provides for the first time an estimate of the total deposits held by Andorra’s banks between 1931 and 2007. Based on this new information, the paper reaches the conclusion that the development of the cluster went through four distinct phases in which large companies, acting as leaders, played an important role in enhancing the cluster’s business capabilities.

Effects of artisanal fisheries on the scorpaenids (Scorpaena spp.) reproduction in the marine protected area of Cap de Creus (NW Mediterranean)

Rockfish species are considered important to the ecology of rocky-reef communities which constitute a key habitat in many coastal marine protected areas (MPAs). These species, which are commercially valuable for artisanal fisheries in the Mediterranean, display particular biological traits that could render them vulnerable to fishing. In this study we monitored the artisanal fisheries of scorpaenids (Scorpaena spp.) in the MPA of Cap de Creus (northwestern Mediterranean) in order to estimate the status of their populations, to link captures with the reproduction of these species, and to evaluate the potential impact that artisanal fishing may have on them. Data from onboard sampling with artisanal fishermen and from fisheries statistics (total landings) were used. Total landings and catch per unit of effort (CPUE) follow a clearly seasonal cycle, with a prominent peak during the summer months coinciding with their spawning season, which may be due to mating behavior prior to fertilization. Although maximum sizes are bigger inside the MPA than in non-protected areas situated close by, a significant percentage of individuals caught inside the MPA are below their size at sexual maturity. Although rockfish seem to be favored by the partial protection of the MPA, the allowed artisanal fisheries are probably impacting the reproduction of these species

Thylakoid Protein Phosphorylation in Regulation of Photosynthesis

Once the seed has germinated, the plant is forced to face all the environmental changes in its habitat. In order to survive, plants have evolved a number of different acclimation systems. The primary reaction behind plant growth and development is photosynthesis. Photosynthesis captures solar energy and converts it into chemical form. Photosynthesis in turn functions under the control of environmental cues, but is also affected by the growth, development, and metabolic state of a plant. The availability of solar energy fluctuates continuously, requiring non-stop adjustment of photosynthetic efficiency in order to maintain the balance between photosynthesis and the requirements and restrictions of plant metabolism. Tight regulation is required, not only to provide sufficient energy supply but also to prevent the damage caused by excess energy. The very first reaction of photosynthesis is splitting of water into the form of oxygen, hydrogen, and electrons. This most fundamental reaction of life is run by photosystem II (PSII), and the energy required for the reaction is collected by the light harvesting complex II (LHCII). Several proteins of the PSII-LHCII complex are reversibly phosphorylated according to the energy balance between photosynthesis and metabolism. Thylakoid protein phosphorylation has been under extensive investigation for over 30 years, yet the physiological role of phosphorylation remains elusive. Recently, the kinases behind the phosphorylation of PSII-LHCII proteins (STN7 and STN8) were identified and the knockout mutants of these kinases became available, providing powerful tools to elucidate the physiological role of PSII-LHCII phosphorylation. In my work I have used the stn7 and stn8 mutants in order to clarify the role of PSII-LHCII phosphorylation in regulation and protection of the photosynthetic machinery according to environmental cues. I show that STN7- dependent PSII-LHCII protein phosphorylation is required to balance the excitation energy distribution between PSII and PSI especially under low light intensities when the excitation energy transfer from LHC to PSII and PSI is efficient. This mechanism differs from traditional light quality-induced “state 1” – “state 2” transition and ensures fluent electron transfer from PSII to PSI under low light, yet having highest physiological relevance under fluctuating light intensity. STN8-dependent phosphorylation of PSII proteins, in turn, is required for fluent turn-over of photodamaged PSII complexes and has the highest importance upon prolonged exposure of the photosynthetic apparatus to excess light.

SYNTHESIS AND ANTIFUNGAL ACTIVITY OF PALMITIC ACID-BASED NEOGLYCOLIPIDS RELATED TO PAPULACANDIN D

A series of six new palmitic acid-based neoglycolipids related to Papulacandin D were synthesized in five steps, resulting in good yields, and they were evaluated against Candida spp. All twelve synthetic intermediates were also evaluated. The synthesis involved the initial glycosylation of two phenols (4-hydroxy-3-methoxybenzaldehyde and 3-hydroxybenzaldehyde) via their reaction with peracetylated glucosyl bromide. This was followed by deacetylation with potassium methoxide/metanol solution and the protection of two hydroxyls (C4 and C6 positions) of the saccharide unit as benzilidene acetals (10-11). The next step involved the acylation of the acetal derivatives with palmitic acid, thereby affording a mixture of two isomers mono-acylated at the C2 and C3 positions and a di-acylated product (12-17). After being isolated, each compound was subjected to the removal of the acetal protecting group to yield the papulacandin D analogues 18-23. Three compounds showed low antifungal activity against two species: C. albicans (compounds 7 and 23) and C. tropicalis (compound 17) at 200 µg mL−1.

SYNTHESIS AND BIOLOGICAL ACTIVITY OF NEW SERIES OF ORGANOTIN(IV) ESTERS WITH N,N-DIACETYLGLYCINE

A bioactive N,N-diacetylglycine (NNDAG) and new organotin(IV) complexes (OTCs) (1-7) were synthesized. Spectroscopic techniques were employed to characterize NNDAG and OTCs. FTIR was employed to verify N,N protection of glycine by acetyl groups. The disappearance of υ(OH) at 3000-2600 cm-1 showed de-protonation of free ligand. The Δυ 150<200 cm-1 of OTCs 4-7 verified bidentate coordination with tetrahedral geometry. The Δυ of OTCs1 and 3 was <200 cm-1 exhibitingtrans -octahedral geometry while OTC 2 dimer was assigned a unique sinusoidal view. The 1H NMR spectra of OTCs verified their synthesis by de-protonation of NNDAG and no chemical shift was found downfield for carboxylic acid proton. The 13C, 119Sn NMR and Mass spectrometric data also supported FTIR and 1H NMR descriptions. The OTCs 4, 5, 6 and7 (500 ppm) proved twice as active against Escherichia coli as the standard antibiotic enoxacin (1000 ppm). The promising property of the OTCs (4, 5, 6 and7) is clearly due to their tetrahedral. The OTCs 4and 5 exhibited excellent activity against M. minimum and good activity against T. castaneum.LD50 of all the compounds were determined and OTCs4, 5 and 7 were found to be active.

Susceptibility of fruits of the 'Valência' and 'Natal' sweet orange varieties to Guignardia citricarpa and the influence of the coexistence of healthy and symptomatic fruits

The importance of the presence of symptomatic fruits with citrus black symptoms (CBS) on the disease severity level in subsequent crop production was assessed in an orchard planted with Valência and Natal sweet orange varieties. Additionally, the period of susceptibility of the fruits of these varieties was evaluated. Fruits were covered with paper bags at the stage of 75% fallen petals and were then exposed to natural infection at weekly intervals, from October 2000 to April 2001. This process was carried out in plants where the fruits from the previous harvest had been picked as well as in plants where fruits remained until natural drop. The evaluation of disease severity used a scale that varied from 0 (absence of symptoms) to 6 (severe symptoms). It was observed that, for the Valência and Natal varieties, conidia of Phyllosticta citricarpa that had formed on the lesions of fruits from the previous harvest did not significantly increase the severity of disease on the fruits of the subsequent harvest period. In this study, the protection of the fruits until 10 weeks after petal drop did not affect the number of lesions, indicating that ascospore discharges after that date were, probably, responsible for disease severity. Fruits exposed between the 20th and 24th week after 75% of the petals had fallen were symptomatic, indicating that, at this stage, the fruits were still susceptible to the pathogen.

Den vårdande och helande bilden: möten med bildkonst i vårdandets värld

Denna vårdvetenskapliga avhandling syftar till att avtäcka och belysa en vårdande och helande dimension vid existentiellt lidande patienters möten med bildkonst inom vårdkontext. Kunskapssökandet sker i två studier. Den första (studie I) är en ikonografi sk tolkning av konstnären Matthias Grünewalds (ca 1460–1528) senmedeltida altarskåpsmålningar. I studien uttolkas lidandets uttryck och narrativa budskap samt symboliska gestaltningar av vårdande och helande i valda delar av detta s.k. Isenheimaltares bildprogram. Tolkningen utgår från rekonstruktionen av altarskåpets ursprungskontext, det medeltida Isenheimklostret, där svårt sjuka och döende patienter vårdades. I studie två (II) fortsätter sökandet i den moderna hospicevårdens kontext med hjälp av en kvalitativ intervjustudie som utforskar patienters meningsskapande vid möten med självvald bildkonst (oljemålningar och akvareller av fi nländska konstnärer som donerats till det sjukhus där intervjustudien gjordes). Forskningsansatsen är inspirerad av Hans-Georg Gadamers (1901–2002) hermeneutik. Vidare används några nyare tolkningsteoretiska ansatser inom bildkonstens område. Forskningens tolkningsresultat visar att bildkonsten har potentialer såväl på ett miljöestetiskt plan som på en djupare individuell symbolnivå. Som designkomponent i vårdmiljöns rumsliga gestaltning bygger bildkonsten in estetiska, etiska och andliga kvaliteter utifrån tidsmässiga och kulturella koder. I den medeltida klostervårdens kontext sammanföll bildkonstens dekorativa betydelse med andliga och helande syften. När det gäller självvalda konstverk i den moderna vårdkontexten bidrar de till det enskilda patientrummets atmosfär på ett unikt sätt utifrån patientens personlighet och behov. På en fördjupad mötesnivå, i samspel med bildens symboliska funktion, sker en inlevelsemässig förfl yttning in i bildens värld. Betraktarens inlevelse aktiveras till en transcenderande rörelse som går bortom det faktiska rummets och den reella tidens gränser. Vid resor i konstens bildvärld spelas minnesvärda händelser upp från det förgångna, men även framtiden kommer betraktaren till mötes. I en existentiell livssituation söker människan i konstverkets bildinnehåll efter symbolisk mening som kan ge svar på lidandets frågor. Bilderna iscensätter då helande motbilder som utgör korrektiv i symboliska former när olika existentiella förluster hotar. När livet förbleknats av sjukdom besvarar bildvärlden den lidandes blick med lysande violer som blommar upp, ger livskraft och bekräftar personens värdighet mitt i det förvissnande människolivet. När ångest och otrygghet nalkas inbjuds betraktaren till besök i landskap som utvidgar sjukhusrummets väggar mot hemgårdens trygghet. Där livet hotas av förgänglighet tar bildvärlden människan med sig till naturens eviga återfödelse. Upplevelsen av att vara delaktig i ett större och heligt sammanhang öppnar vägen ut ur lidandets avskurenhet. I medeltidens vårdkontext erbjöd den sakrala bilden en kollektiv och helande Symbolon som genom sin representationskraft synliggjorde det osynliga. Vid bildmöten i den moderna hospicevårdens kontext var det naturteman som gläntade på dörren till ”det hemliga rummet i djupet av hjärtat”. Forskningen antyder att även om meningsskapandet i ett bildmöte är avhängigt tidsepok, betraktarens förförståelse och kulturella kontext samt typen av bilder kan bildsymboliken, generellt förstådd som den saknade formen eller det saknade livssammanhanget, framvisa en helande och hoppingivande ordning i lidandets kaos.

Efficiency of a yeast-based formulation for the biocontrol of postharvest anthracnose of papayas

To identify formulations of biological agents that enable survival, stability and a good surface distribution of the antagonistic agent, studies that test different application vehicles are necessary. The efficiency of two killer yeasts, Wickerhamomyces anomalus (strain 422) and Meyerozyma guilliermondii (strain 443), associated with five different application vehicles, was assessed for the protection of postharvest papayas. In this study, after 90 days of incubation at 4ºC, W. anomalus (strain 422) and M. guilliermondii (strain 443) were viable with all application vehicles tested. Fruits treated with different formulations (yeasts + application vehicles) had a decreased severity of disease (by at least 30%) compared with untreated fruits. The treatment with W. anomalus (strain 422) + 2% starch lowered disease occurrence by 48.3%. The most efficient treatments using M. guilliermondii (strain 443) were those with 2% gelatin or 2% liquid carnauba wax, both of which reduced anthracnose by 50% in postharvest papayas. Electron micrographs of the surface tissues of the treated fruits showed that all application vehicles provided excellent adhesion of the yeast to the surface. Formulations based on starch (2%), gelatin (2%) and carnauba wax (2%) were the most efficient at controlling fungal diseases in postharvest papayas.

Formal and informal mechanisms for knowledge protection and sharing

The most important knowledge in firms is mostly tacit and embedded in individuals within the organization. This background knowledge that firms possess is used for creation of new knowledge and innovations. As firms today greatly concentrate on their core competencies, they need external knowledge from various collaboration partners. Thus, collaborative relationship governance, as well as control (use of appropriability mechanisms) over background (the input from each firm in innovative activities) and foreground knowledge (the output of collaboration activities) is needed in order to successfully create and capture value from innovative activities without losing core knowledge and competitiveness. Even though research has concentrated on knowledge protection and knowledge sharing, studies that combine both of these views and examine the effects of sharing and protection on value creation and capture have been rather limited. Studies have mainly focused on the protection of the output of innovation while forgetting the protection of the input of innovation. On the other hand, as the research concentrating on the output of innovation tends to favor formal mechanisms, informal mechanisms have remained more unknown to researchers as well as managers. This research aims to combine the perspectives of knowledge sharing and knowledge protection and their relationship with value creation and value capture. The sharing and protection are viewed from two points of view: the use of appropriability mechanisms, as well as governance of the collaborative relationship. The study consists of two parts. The first part introduces the research topic and discusses the overall results. The second part comprises six complementary research publications. Both qualitative and quantitative research methods are used in the study. In terms of results, the findings enhance understanding of the combined use of formal and informal mechanisms for knowledge protection and sharing. Informal mechanisms appear to be emphasized in the protection of background knowledge, and thus are prerequisites for innovation, whereas formal mechanisms are relied on more for protecting the results of innovative activities. However, the simultaneous use of the formal and informal mechanisms that are relevant to the particular industry and innovation context is recommendedthroughout the collaborative innovation process. Further, the study adds to the current knowledge on HRM as an appropriability mechanism: on the firm level its uses include assessing and hedging against employee-related risks such as knowledge leaking and knowledge leaving. A further contribution is to the research on HRM protection and its interrelations with other appropriability mechanisms, its constituents, and its potential use in the area of knowledge protection.

Application of Open Innovation Paradigm in Small and Medium Enterprises in Russia

The objective of this study is to explore how the Open Innovation paradigm is applied in by small and medium-size enterprises in Russia. The focus of the study is to understand how the processes of research and development and commercialization proceed in these kind of companies and to which extent they apply open innovation principles. Russian leadership makes certain steps for transition from the export of raw materials to an innovative model of economic growth. The research aims to disclose actual impact of these attempts. The closed innovation model and the erosion factors which lead to the destruction of an old one and emergence of new model are described. Features of open innovation implementation and intellectual property rights protection in small and medium enterprises are presented. To achieve the objective, a qualitative case study approach was chosen. Research includes facts and figures, views and opinions of management of studied companies related to innovation process in the company and in Russia in general. The research depicts the features of Open Innovation implementation by SMEs in Russia. A large number of research centers with necessary equipment and qualified personnel allow case companies to use external R&D effectively. They cooperate actively with research institutes, universities and laboratories. Thus, they apply inbound Open Innovation. On the contrary, lack of venture capital, low demand for technologies within the domestic market and weak protection of intellectual property limit the external paths to new markets. Licensing-out and creation of spin-off are isolated cases. Therefore, outbound Open Innovation is not a regular practice.

Reglering och förvaltning av produktions- och slaktdjurs välbefinnande : en offentligrättslig undersökning

Djur och djurskyddet hör till ett rättsligt regleringsområde som inte varit föremål för omfattande rättsvetenskaplig forskning i vårt land även om intresset för djurs välbefinnande ökat både i samhället och inom EU. Avhandlingen har avfattats som en artikelavhandling och är en offentligrättslig studie om djurskyddslagstiftningen och förvaltningen av djurskyddsärenden i Finland. Tematiken har behandlats både ur djurens och djurens ägares eller innehavares synvinkel med utgångspunkt i djurskyddslagstiftningen och förvaltningen av djurskyddsärenden i Finland. Forskningen är fokuserad huvudsakligen på skyddet av och välbefinnandet hos produktions- och slaktdjur även om bland annat de begrepp som granskas också berör andra djurkategorier. De övergripande frågeställningarna i avhandlingen är två. För det första, vad är det som avses med djurs välbefinnande och skydd i regleringen av djurskyddet och för det andra, hur realiseras dessa i djurskyddsmyndigheternas förvaltningsverksamhet? I forskningen presenteras och diskuteras bland annat en ny begreppskonstruktion: djurs rättsliga välbefinnande. Den empiriska delen i avhandlingen omfattar förvaltningsverksamheten inom området av djurskydd under åren 1996–2006. Sammanlagt 10468 dokument som upprättats av djurskyddsmyndigheten i samband med verkställandet av djurskyddsövervakning ingår i undersökningen. Forskningen utmynnar i en åtgärdsförteckning med förslag till utvecklingen av området för djurskydd.

Granulocyte-macrophage colony-stimulating factor does not increase the potency or efficacy of a foot-and-mouth disease virus subunit vaccine

Foot-and-mouth disease (FMD) is one of the most feared diseases of livestock worldwide. Vaccination has been a very effective weapon in controlling the disease, however a number of concerns with the current vaccine including the inability of approved diagnostic tests to reliably distinguish vaccinated from infected animals and the need for high containment facilities for vaccine production, have limited its use during outbreaks in countries previously free of the disease. A number of FMD vaccine candidates have been tested and a replication-defective human adenovirus type 5 (Ad5) vector containing the FMDV capsid (P1-2A) and 3C protease coding regions has been shown to completely protect pigs against challenge with the homologous virus (FMDV A12 and A24). An Ad5-P1-2A+3C vaccine for FMDV O1 Campos (Ad5-O1C), however, only induced a low FMDV-specific neutralizing antibody response in swine potency tests. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been successfully used to stimulate the immune response in vaccine formulations against a number of diseases, including HIV, hepatitis C and B. To attempt to improve the FMDV-specific immune response induced by Ad5-O1C, we inoculated swine with Ad5-O1C and an Ad5 vector containing the gene for porcine GM-CSF (pGM-CSF). However, in the conditions used in this trial, pGM-CSF did not improve the immune response to Ad5-O1C and adversely affected the level of protection of swine challenged with homologous FMDV.