Mechanisms of KCC2 upregulation during development

Gamma-aminobutyric acid (GABA) acting through ionotropic GABAA receptors plays a crucial role in the activity of the central nervous system (CNS). It triggers Ca2+ rise providing trophic support in developing neurons and conducts fast inhibitory function in mature neuronal networks. There is a developmental change in the GABAA reversal potential towards more negative levels during the first two postnatal weeks in rodent hippocampus. This change provides the basis for mature GABAergic activity and is attributable to the developmental expression of the neuron-specific potassium chloride cotransporter 2 (KCC2). In this work we have studied the mechanisms responsible for the control of KCC2 developmental expression. As a model system we used hippocampal dissociated cultures plated from embryonic day (E) 17 mice embryos before the onset of KCC2 expression. We showed that KCC2 was significantly up-regulated during the first two weeks of culture development. Interestingly, the level of KCC2 upregulation was not altered by chronic pharmacological blockage of action potentials as well as GABAergic and glutamatergic synaptic transmission. By in silico analysis of the proximal KCC2 promoter region we identified 10 candidate transcription factor binding sites that are highly conserved in mammalian KCC2 genes. One of these transcription factors, namely early growth response factor 4 (Egr4), had similar developmental profile as KCC2 and considerably increased the activity of mouse KCC2 gene in neuronal cells. Next we investigated the involvement of neurotrophic factors in regulation of Egr4 and KCC2 expression. We found that in immature hippocampal cultures Egr4 and KCC2 levels were strongly up-regulated by brain derived neurotrophic factor (BDNF)and neurturin. The effect of neurotrophic factors was dependent on the activation of a mitogen activated protein kinase (MAPK) signal transduction pathway. Intact Egr4-binding site in proximal KCC2 promoter was required for BDNF-induced KCC2 transcription. In vitro data were confirmed by several in vivo experiments where we detected an upregulation of KCC2 protein levels after intrahippocampal administration of BDNF or neurturin. Importantly, a MAPK-dependent rise in Egr4 and KCC2 expression levels was also observed after a period of kainic acid-induced seizure activity in neonatal rats suggesting that neuronal activity might be involved in Egr4-mediated regulation of KCC2 expression. Finally we demonstrated that the mammalian KCC2 gene (alias Slc12a5) generated two neuron-specific isoforms by using alternative promoters and first exons. A novel isoform of KCC2, termed KCC2a, differed from the previously known KCC2b isoform by 40 unique N-terminal amino acid residues. KCC2a expression was restricted to CNS,remained relatively constant during postnatal development, and contributed 20 50% of total KCC2 mRNA expression in the neonatal mouse brainstem and spinal cord. In summary, our data provide insight into the complex regulation of KCC2 expression during early postnatal development. Although basal KCC2 expression seems to be intrinsically regulated, it can be further augmented by neurotrophic factors or by enhanced activity triggering MAPK phosphorylation and Egr4 induction. Additional KCC2a isoform, regulated by another promoter, provides basal KCC2 level in neonatal brainstem and spinal cord required for survival of KCC2b knockout mice.

Talouskäsitysten vaikutus hallituksen suosioon Puolassa 2006–2007

Tutkimuksessa tarkastellaan äänestäjien talouskäsitysten vaikutusta hallituksen suosioon Puolassa vuosina 2006 ja 2007. Tutkimusasetelma juontaa juurensa poliittisten suhdannevaihteluiden teoriaan, jonka mukaan poliitikot pyrkivät manipuloimaan taloutta vaalien alla tullakseen uudelleen valituiksi. Vastuullisuushypoteesin mukaan äänestäjät palkitsevat poliitikot hyvästä taloustilanteesta. Näin ollen mikäli äänestäjät ovat tyytyväisiä talouteen he äänestävät hallitusta. Mikäli eivät, he äänestävät oppositiota. Miten äänestäjät sitten tekevät arvionsa taloudesta? Tutkielmassa selvitetään vaikuttiko äänestäjien mielipiteisiin käsitys nykyisestä vai tulevaisuuden taloustilanteesta. Käsitys nykyisestä taloustilanteesta perustuu havaintoihin jo toteutuneesta talouspolitiikasta. Tätä käsitystä kutsutaan retrospektiiviseksi näkemykseksi. Prospektiiviset talousnäkemykset koostuvat näkemyksestä ja odotuksista talouden tulevasta tilasta ja uskosta hallituksen kykyyn tarjota paras vaihtoehto suotuisalle talouskehitykselle. Aikaulottuvuuden lisäksi tutkielmassa testataan, vaikuttaako äänestäjiin enemmän heidän oma taloustilanteensa vai näkemys yleisestä talouden tilanteesta. Mikäli hallituksen suosio perustuu näkemyksiin henkilökohtaisen talouden tilasta, ovat äänestäjät egotrooppisia. Mikäli kannatuspäätös perustuu näkemykseen kansantalouden tilasta, ovat äänestäjät puolestaan sosiotrooppisia. Aineistona tutkielmassa käytetään uusinta saatavilla olevaa Puolan mielipidetiedustelun (Centrum Badania Opinii Publicznej) keräämää kyselyaineistoa vuosilta 2006 ja 2007. Tutkielmassa on yhdistetty kyselyaineistot 22:n kuukauden ajalta. Aineistoa on analysoitu logistisen regressioanalyysin menetelmin. Tutkielman perusteella talousnäkemyksistä suurin vaikutus hallituksen suosioon on äänestäjien näkemyksillä kansantalouden tulevaisuuden tilasta. Henkilökohtaisen talouden osalta positiiviset tulevaisuudennäkymät vaikuttavat positiivisesti myös hallituksen kannatukseen. Retrospektiivinen arvio henkilökohtaisesta taloudesta toimii kuitenkin teorian vastaisesti; oman taloudellisen tilanteen hyväksi arvioineiden todennäköisyys kannatta hallitusta oli pienempää kuin niillä, jotka arvioivat oman taloustilanteensa huonoksi. Kontrollimuuttujiksi valittiin joukko aiemmassa tutkimuksessa äänestyskäyttäytymiseen Puolassa vaikuttaneita tekijöitä. Näitä olivat ikä, sukupuoli, koulutus, asuinpaikka, näkemys työllisyystilanteesta, ideologinen sijoittuminen oikeisto–vasemmisto-akselilla sekä uskonnollisuus. Kontrollimuuttujista merkitseviksi osoittautuivat uskonnollisuus, oikeisto–vasemmisto-ideologia, koulutus, asuinpaikka sekä ikä. Kun edellä mainitut taustamuuttujat on kontrolloitu, taloustekijät selittävät noin viidesosan hallituksen kannatuksen todennäköisyyden vaihtelusta.

The relationships between work stressors and organizational performance in long-term care for elderly residents

This study investigates the relationships between work stressors and organizational performance in terms of the quality of care provided by the long-term care facilities. Work stressors are first examined in relation to the unit's structural factors, resident characteristics, and to the unit specialization. The study is completed by an investigation into the associations of work stressors such as job demands or time pressure, role ambiguity, resident-related stress, and procedural injustice to organizational performance. Also the moderating effect of job control in the job demands organizational performance relationship is examined. The study was carried out in the National Research and Development Centre for Welfare and Health (STAKES). Survey data were drawn from 1194 nursing employees in 107 residential-home and health-center inpatient units in 1999 and from 977 employees in 91 units in 2002. Information on the unit resident characteristics and the quality of care was provided by the Resident Assessment Instrument (RAI). The results showed that large unit size or lower staffing levels were not consistently related to work stressors, whereas the impairments in residents' physical functioning in particular initiated stressful working conditions for employees. However, unit specialization into dementia and psychiatric residents was found to buffer the effects that the resident characteristics had on employee appraisals of work stressors, in that a high proportion of behavioral problems was related to less time pressure and role conflicts for employees in specialized units. Unit specialization was also related to improved team climates and the organizational commitment of employees. Work stressors associated with problems in care quality. Time pressure explained most of the differences between units in how the employees perceived the quality of physical and psychosocial care they provide for the residents. A high level of job demands in the unit was also found to be related to some increases in all clinical quality problems. High job control buffered the effects of job demands on the quality of care in terms of the use of restraints on elderly residents. Physical restraint and especially antipsychotic drug use were less prevalent in units that combined both high job demands and high control for employees. In contrast, in high strain units where heavy job demands coincided with a lack of control for employees, quality was poor in terms of the frequent use of physical restraints. In addition, procedural injustice was related to the frequent use of antianxiety of hypnotic drugs for elderly residents. The results suggest that both job control and procedural justice may have improved employees' abilities to cope when caring for the elderly residents, resulting in better organizational performance.

Widening Perspectives of Teacher Education : Studies on theory-practice relationship, reflection, research and professional development

This dissertation examined the research-based teacher education at the University of Helsinki from different theoretical and practical perspectives. Five studies focused on these perspectives separately as well as overlappingly. Study I focused on the reflection process of graduating teacher students. The data consisted of essays the students wrote as their last assignment before graduating, where their assignment was to examine their development as researchers during their MA thesis research process. The results indicated that the teacher students had analysed their own development thoroughly during the process and that they had reflected on theoretical as well as practical educational matters. The results also pointed out that, in the students’ opinion, personally conducted research is a significant learning process. -- Study II investigated teacher students’ workplace learning and the integration of theory and practice in teacher education. The students’ interviews focused on their learning of teacher’s work prior to education. The interviewees’ responses concerning their ‘surviving’ in teaching prior to teacher education were categorized into three categories: learning through experiences, school as a teacher learning environment, and case-specific learning. The survey part of the study focused on integration of theory and practice within the education process. The results showed that the students who worked while they studied took advantage of the studies and applied them to work. They set more demanding teaching goals and reflected on their work more theoretically. -- Study III examined practical aspects of the teacher students’ MA thesis research as well as the integration of theory and practice in teacher education. The participants were surveyed using a web-based survey which dealt with the participants’ teacher education experiences. According to the results, most of the students had chosen a practical topic for their MA thesis, one arising from their work environment, and most had chosen a research topic that would develop their own teaching. The results showed that the integration of theory and practice had taken place in much of the course work, but most obviously in the practicum periods, and also in the courses concerning the school subjects. The majority felt that the education had in some way been successful with regards to integration. -- Study IV explored the idea of considering teacher students’ MA thesis research as professional development. Twenty-three teachers were interviewed on the subject of their experiences of conducting research about their own work as teachers. The results of the interviews showed that the reasons for choosing the MA thesis research topic were multiple: practical, theoretical, personal, professional reasons, as well as outside effect. The objectives of the MA thesis research, besides graduating, were actual projects, developing the ability to work as teachers, conducting significant research, and sharing knowledge of the topic. The results indicated that an MA thesis can function as a tool for professional development, for example in finding ways for adjusting teaching, increasing interaction skills, gaining knowledge or improving reflection on theory and/or practice, strengthening self-confidence as a teacher, increasing researching skills or academic writing skills, as well as becoming critical and being able to read scientific and academic literature. -- Study V analysed teachers’ views of the impact of practitioner research. According to the results, the interviewees considered the benefits of practitioner research to be many, affecting teachers, pupils, parents, the working community, and the wider society. Most of the teachers indicated that they intended to continue to conduct research in the future. The results also showed that teachers often reflected personally and collectively, and viewed this as important. -- These five studies point out that MA thesis research is and can be a useful tool for increasing reflection doing with personal and professional development, as well as integrating theory and practice. The studies suggest that more advantage could be taken of the MA thesis research project. More integration of working and studying could and should be made possible for teacher students. This could be done in various ways within teacher education, but the MA thesis should be seen as a pedagogical possibility.

Literaturnaja skazka A.M. Remizova (1900-1920-e gody)

Literary tale of A.M. Remizov (1900’s – 1920’s) The thesis is devoted to a detailed historical-literary description of a tale as a genre tradition in the creative work of Alexei Mikhailovich Remizov (1877-1957), one of the major Russian prose writers of the 20’s century. This very approach allows to specify the place and functional meaning of this genre in literary practice of the writer and to appeal to one of the key problems of the 20-21 century literature history – a specific of modernistic literature composition principle and a role of montage techniques in its formation. Remizov was working on tales during his whole life, though the most productive years of folklore studies fell to 1900’s – 1910’s. During this period he intensively studied folklore materials, narrated several hundreds of folk tales and in 1900’s – 1920’s published eight tale collections which played a significant role in the formation of stylistic and compositional principles of his prose of the 1910’s – 1920’s, especially montage techniques, which in its turn influenced the development of the narrative forms in the Russian post-revolutionary literature. At the same time a tale has specified not only poetics but also problematics of Remizov’s creative work, as when choosing folklore sources the writer always alluded to modern themes and relevant intellectual trends. The current research work, based on various archive materials and a wide spectrum of modern historical-literary data, complies four chapters with a consistent description of creation history, publication and critics’ reviews of Remizov’s tale collections and single tales contributing to his creative evolution characteristic. Furthermore, the work refers to composition and subject of the particular collections. On the whole it enables to follow up genre dynamics. The first chapter of the work is devoted to Posolon’ (Sunwise), the earliest tale collection of Remizov. The main feature of the collection is that its composition is oriented on the agrarian calendar and the subject – on the system of mythological views reflected in the Russian folklore. This very collection to a large extent corresponds to the writer’s views on the myth represented in Pis’mo v redaktsiyu (Letter to the Editor). The history of this manifesto appearing is analyzed in the second chapter. The incident which caused its forthcoming contributed to ‘legitimization’ of Remizov’s narrations as a relevant genre of modern literature and to upgrading the writer in professional hierarchy. The third chapter analyzes Remizov’s collections of 1900’s – early 1920’s, a result of Remizov’s scrupulous work with a specific tale material. He is acting here as a tale repertory researcher and in some cases as a collector as well. The means of such collections’ topical organization is not the myth but the hero of the tale. According to this principle single pieces are grouped into cycles, which then form complicated montage constructs. Texts themselves can be viewed as a sort of hyper-quotations, as they in fact entirely coincide with their original sources. Besides, collections usually have their own ideal patterns. In the fourth chapter a connection of Remizov’s creative work with folk fun culture and a tradition of the folklore noel story is being demonstrated on Zavetnyie skazy (Secret Tales) material. A consistent collections’ history creation analysis convinces us that the tale was a sort of laboratory in which main writer’s prose methods were being worked out.

The Europeanisation of occupational health services: A study of the impact of EU policies

This study concerns Framework Directive 89/391/EEC on health and safety at work, which encouraged improvements in occupational health services (OHS) for workers in EU member states. Framework Directive 89/391/EEC originally aimed at bringing the same level of occupational health and safety to employees in both the public and private sectors in EU member states. However, the implementation of the framework directive and OHS varies widely among EU member states. Occupational health services have generally been considered an important work-related welfare benefit in EU member states. The purpose of this study was to analyse OHS within the EU context and then analyse the impact of EU policies on OHS implementation as part of the welfare state benefit. The focus is on social, health, and industrial policies within welfare state regimes as well as EU policy-making processes affecting these policies in EU member states. The research tasks were divided into four groups related to the policy, functions, targets,and actors of OHS. The questions related to policy tried to discover the role of OHS in other policies, such as health, social, and labour market policies within the EU. The questions about functions sought to describe the changes, as well as the path dependence, of OHS in EU member states after the framework directive. The questions about targets were based on the general aims of WHO and the ILO in relation to equity, solidarity, universality, and access to OHS. The questions on actors were designed to understand the variety of stakeholders interested in OHS. The actors were supranational (EU, ILO, and WHO), national (ministries, institutes, and professional organisations), and social partners (trade unions and employers organisations). The study data were collected by interviewing 92 people in 15 EU member states, including representatives of ministries, institutions, research,trade unions, employers organisations, and occupational health organisations. Other documents were collected from the Internet,databases, libraries, and conference materials for a systematic review of the policies, strategies, organisation, financing, and monitoring of OHS in EU member states. Different analytical methods were used in the data analysis. The main findings of the study can be summarised as follows. First, occupational health services is a context-dependent phenomenon, which therefore varies according to the development of the welfare state in general, and depends on each country s culture, history, economy, and politics. The views of different stakeholders in EU member states concerning the impact and possibilities of OHS to improve health vary from evidence-based opinions to the sporadic impact of OHS on occupational health. OHS as a concept is vaguely defined by the EU, whereas the ILO defines OHS content. The tasks of OHS began as preventive and protective services for workers. However, they have moved towards multidisciplinary and organisational development as well as the workplace health promotion sphere.Since 1989 OHS has developed differently in different EU member states depending on the starting position of those states, but planning and implementation are crucial phases in the process toward better OHS coverage, equity, and access. Nevertheless, the data used for the planning and legitimisation of OHS activities are mainly based on occupational health data rather than on OHS data. This makes decisions on political or policy grounds inaccurate. OHS is still an evolving concept and benefit for workers, but the Europeanisation of OHS reflects contextual changes, such as the impact of the internal market, competition, and commercialisation on OHS. Stronger cooperation and integration with health, social, and employment services would be an asset for workers, because of new epidemics, an epidemiological shift towards new risks, an ageing labour market, and changes in the labour market. Different methods and approaches are needed in order to study the results of integrated services. In the future, more detailed information will be needed about the actual impact of EU policies on OHS and decision-making processes in order to get OHS into different policies in the EU and its member states. Further results and effects of OHS processes on occupational health need to be analysed more carefully. The adoption of a variety of research strategies and a multidisciplinary approach to understand the influence of different policies on OHS in the EU and its member states would highlight the options and opportunities to improve workers occupational health. Key subject headings: Occupational health services, EU policy, policymaking,framework directive 89/391/EEC

Beasts on Fields. Human-Wildlife Conflicts in Nature-Culture Borderlands

Human-wildlife conflicts are today an integral part of the rural development discourse. In this research, the main focus is on the spatial explanation which is not a very common approach in the reviewed literature. My research hypothesis is based on the assumption that human-wildlife conflicts occur when a wild animal crosses a perceived borderline between the nature and culture and enters into the realms of the other. The borderline between nature and culture marks a perceived division of spatial content in our senses of place. The animal subject that crosses this border becomes a subject out of place meaning that the animal is then spatially located in a space where it should not be or where it does not belong according to tradition, custom, rules, law, public opinion, prevailing discourse or some other criteria set by human beings. An appearance of a wild animal in a domesticated space brings an uncontrolled subject into that space where humans have previously commanded total control of all other natural elements. A wild animal out of place may also threaten the biosecurity of the place in question. I carried out a case study in the Liwale district in south-eastern Tanzania to test my hypothesis during June and July 2002. I also collected documents and carried out interviews in Dar es Salaam in 2003. I studied the human-wildlife conflicts in six rural villages, where a total of 183 persons participated in the village meetings. My research methods included semi-structured interviews, participatory mapping, questionnaire survey and Q- methodology. The rural communities in the Liwale district have a long-history of co-existing with wildlife and they still have traditional knowledge of wildlife management and hunting. Wildlife conservation through the establishment of game reserves during the colonial era has escalated human-wildlife conflicts in the Liwale district. This study shows that the villagers perceive some wild animals differently in their images of the African countryside than the district and regional level civil servants do. From the small scale subsistence farmers point of views, wild animals continue to challenge the separation of the wild (the forests) and the domestics spaces (the cultivated fields) by moving across the perceived borders in search of food and shelter. As a result, the farmers may loose their crops, livestock or even their own lives in the confrontations of wild animals. Human-wildlife conflicts in the Liwale district are manifold and cannot be explained simply on the basis of attitudes or perceived images of landscapes. However, the spatial explanation of these conflicts provides us some more understanding of why human-wildlife conflicts are so widely found across the world.

Kansallisen konsensuskulttuurin jäljillä : Globalisaatioajan Suomi Helsingin Sanomissa

The dissertation analyzes Finnish consensual culture in public discussion and journalism in Helsingin Sanomat (HS). The consensual Finnish political culture has evolved and persisted over a long period of time and it has been affected by historical circumstances as well as the dynamics of political and journalistic structures and actors. A historical chronology is drawn in the study regarding the nature and development of consensus culture in 20th century Finland. This political culture is traced by looking at public discussion on globalization at the turn of the millennium. Globalization as a concept has been contested and various societal actors have given different meanings to it. This research looks at how the globalization discussion in HS during the years 1992-2004 constructs consensus. Helsingin Sanomat (and its predecessor Päivälehti) has been an important actor in Finnish journalism and the public sphere almost since its founding 120 years ago. The history of the paper is tightly connected to Finland s general political history and history of the public sphere. Moreover, the paper s connections to the societal elite have always been close. The central question in this research was to see how the globalization discussion in HS evolved in relation to consensus as well as legitimate controversies. As a result it is stated that the globalization question has clearly divided the Finnish societal actors. The most powerful societal elites (government, most civil servants, corporate sector) had a profile of being pro globalization. They communicated their globalization strategy as a national, unified way of thinking. Other elites which have been losing their influence (the president, labor union, part of members of parliament), as well as civil society actors tried to bring forward conflicting views in relation to globalization. The paper did give some room to these elements, but on the other hand it also tried to keep up the consensual discussion culture especially in the editorial section. In line with its traditions Helsingin Sanomat strived to create national unity. At the same time it did not give adequate attention to the changes brought about by globalization to the positions and roles of various elites and civil society actors. In this discussion HS seemed more like a medium of the state than as a critical and independent actor. Journalism has an important role in upholding and also reviving the Finnish political culture and public discussion. From this point of view it is problematic if the area of so called legitimate controversy in broad societal questions like globalization becomes very limited. As the Finnish elites are small and there is no considerable competition between them, journalism should actively bring up controversial issues. This task becomes complicated, however, if the elite circles are closed up and no initiatives come from their ranks. Political decision making as well as democracy can suffer, if issues are not brought to the public agenda.

Working with Identities Promoting Student Teachers Professional Development

The starting point of the study was that good teaching requires a teacher to be aware of the sources of his or her pedagogical decisions; that is, personal values, beliefs and understandings. From this perspective, a teacher s professional development refers to a process of extending one s self-knowledge. The aim of this study was to promote student teachers' professional development with the help of identity work. Identity work refers to reflecting on both personal and professional experiences. Identity work consists of student teachers self-reflection on their life experiences (self-identity) and video diary-based reflection on their classroom practice (professional identity). The research questions of the study were 1) how is identity work manifested by the participants? and 2) what is the potential of identity work in promoting student teachers professional development? The research data was collected from four student teachers in the academic year 2007-2008; the research group had 24 meetings during the research process. Student teachers take part in a multimode teacher education programme, where students work as ordinary teachers in schools during their university studies. The data collection was conducted by using two methods: participants narratives based on autobiographical writings and video diaries based on participants lessons. Narrative research data was analysed by employing qualitative methods and strategies as they were needed in the research. The research results revealed four different ways of working with identities, each of them revealing different aspects of and approaches to identity work. The results also showed that identity work has the potential to promote professional development. As the research progressed, there were visible changes in the participants reflection. However, despite encouraging results, some issues should be critically questioned. Although reflection sounds attractive and fruitful as a tool for promoting professional development, there are also difficulties and obstacles. On the basis of the results, a proposal for promoting student teachers professional development is offered. Keywords: Teacher identity, identity work, reflection, teacher professional development

Information Technology Development Needs in Community Pharmacies : A Strategic Approach

In the context of health care, information technology (IT) has an important role in the operational infrastructure, ranging from business management to patient care. An essential part of the system is medication management in inpatient and outpatient care. Community pharmacists strategy has been to extend practice responsibilities beyond dispensing towards patient care services. Few studies have evaluated the strategic development of IT systems to support this vision. The objectives of this study were to assess and compare independent Finnish community pharmacy owners and staff pharmacists priorities concerning the content and structure of the next generation of community pharmacy IT systems, to explore international experts visions and strategic views on IT development needs in relation to services provided in community pharmacies, to identify IT innovations facilitating patient care services and to evaluate their development and implementation processes, and to assess community pharmacists readiness to adopt innovations. This study applied both qualitative and quantitative methods. A qualitative personal interview of 14 experts in community pharmacy services and related IT from eight countries and a national survey of Finnish community pharmacy owners (mail survey, response rate 53%, n=308), and of a representative sample of staff pharmacists (online survey, response rate 22%, n=373) were conducted. Finnish independent community pharmacy owners gave priority to logistical functions but also to those related to medication information and patient care. The managers and staff pharmacists have different views of the importance of IT features, reflecting their different professional duties in the community pharmacy. This indicates the need for involving different occupation groups in planning the new IT systems for community pharmacies. A majority of the international experts shared the vision of community pharmacy adopting a patient care orientation; supported by IT-based documentation, new technological solutions, access to information, and shared patient data. Community pharmacy IT innovations were rare, which is paradoxical because owners and staff pharmacists perception of their innovativeness was seen as being high. Community pharmacy IT systems development processes usually had not undergone systematic needs assessment research beforehand or evaluation after the implementation and were most often coordinated by national governments without subsequent commercialization. Specifically, community pharmacy IT developments lack research, organization, leadership and user involvement in the process. Those responsible for IT development in the community pharmacy sector should create long-term IT development strategies that are in line with community pharmacy service development strategies. This could provide systematic guidance for future projects to ensure that potential innovations are based on a sufficient understanding of pharmacy practice problems that they are intended to solve, and to encourage strong leadership in research, development of innovations so that community pharmacists potential innovativeness is used, and that professional needs and strategic priorities will be considered even if the development process is led by those outside the profession.

Development of tissue culture and virus-induced gene silencing for Calendula officinalis

Calendula officinalis is grown widely as an ornamental plant across Europe. It belongs to the large. Asteraceae family. In this study, the aim was to explore the possibilities to use Calendula officinalis as a new model organism for flower development and secondary mechanism studies in Asteraceae. Tissue culture of Calendula officinalis was established using nine different cultivars. Murashige & Skoog (MS) medium with four different combinations of plant growth regulators were tested. Of all these combinations, the medium containing 1mg/l BAP, 0.1 mg/l IAA, and 1mg/l Zeatin achieved highest frequency of adventitious shoot regeneration from hypocotyl and cotyledon explants. Virus-induced gene silencing is a recent developed genetic tool for charactering the gene functions in plants, and extends the range of host plants that are not accessible for Agrobacterium transformation. Here, tobacco rattle virus (TRV)-based VIGS technique was tested in calendula (cv. Single Orange). We used TRV carrying Gerbera hybrid phytoene desaturase (PDS) gene fragment to induce PDS silencing in calendula. Vacuum infiltration and syringe infiltration methods both resulted in photo-bleaching phenotypes in leaves, bracts and petals. Loss-of-function phenotypes occurred on calendula 13 days post-infiltration. In conclusion, the data indicates that calendula explants can be regenerated through tissue culture which is a prerequisite for development of stable transformation methods. However, further optimization is still needed to improve the frequency. In addition, VIGS was applied to silence PDS marker gene expression indicating that this method has potential for gene functional studies in future.