8 resultados para Culture Media, Conditioned

em Doria (National Library of Finland DSpace Services) - National Library of Finland, Finland


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Monocytes, macrophages and dendritic cells (DCs) are important mediators of innate immune system, whereas T lymphocytes are the effector cells of adaptive immune responses. DCs play a crucial role in bridging innate and adaptive immunity. Naïve CD4+ Th progenitors (Thp) differentiate to functionally distinct effector T cell subsets including Th1, Th2 and Th17 cells, which while being responsible for specific immune functions have also been implicated in pathological responses, such as autoimmunity, asthma and allergy. The main objective of this thesis is to dissect the signalling networks involved in the IL-4 induced differentiation of two important leukocyte subtypes, Th2 cells and DCs. Gene expression profiling lead to identification of over 200 genes which are differentially expressed during cytokine induced differentiation of human monocytes to DCs or macrophages and which are likely to be essential for the proper biological functions of these cell types. Transcriptome analysis demonstrated the dynamic regulation of gene expression by IL-12 and IL-4 during the initiation of Th cell differentiation, which was partly counteracted by an immunosuppressive cytokine, TGFβ, present in the culture media. Results from RNAi mediated gene knockdown experiments and global gene expression analysis elucidated that SATB1 regulates multiple genes important for Th cell polarization or function as well as may compete with GATA3 for the reciprocal regulation of IL-5 transcription. In conclusion, the results obtained have extended our system-level understanding of the immune cell differentiation processes and provide an excellent basis for the further functional studies which could lead to development of improved therapeutic approaches for a range of immunological conditions.

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The number of molecular diagnostic assays has increased tremendously in recent years.Nucleic acid diagnostic assays have been developed, especially for the detection of human pathogenic microbes and genetic markers predisposing to certain diseases. Closed-tube methods are preferred because they are usually faster and easier to perform than heterogenous methods and in addition, target nucleic acids are commonly amplified leading to risk of contamination of the following reactions by the amplification product if the reactions are opened. The present study introduces a new closed-tube switchable complementation probes based PCR assay concept where two non-fluorescent probes form a fluorescent lanthanide chelate complex in the presence of the target DNA. In this dual-probe PCR assay method one oligonucleotide probe carries a non-fluorescent lanthanide chelate and another probe a light absorbing antenna ligand. The fluorescent lanthanide chelate complex is formed only when the non-fluorescent probes are hybridized to adjacent positions into the target DNA bringing the reporter moieties in close proximity. The complex is formed by self-assembled lanthanide chelate complementation where the antenna ligand is coordinated to the lanthanide ion captured in the chelate. The complementation probes based assays with time-resolved fluorescence measurement showed low background signal level and hence, relatively high nucleic acid detection sensitivity (low picomolar target concentration). Different lanthanide chelate structures were explored and a new cyclic seven dentate lanthanide chelate was found suitable for complementation probe method. It was also found to resist relatively high PCR reaction temperatures, which was essential for the PCR assay applications. A seven-dentate chelate with two unoccupied coordination sites must be used instead of a more stable eight- or nine-dentate chelate because the antenna ligand needs to be coordinated to the free coordination sites of the lanthanide ion. The previously used linear seven-dentate lanthanide chelate was found to be unstable in PCR conditions and hence, the new cyclic chelate was needed. The complementation probe PCR assay method showed high signal-to-background ratio up to 300 due to a low background fluorescence level and the results (threshold cycles) in real-time PCR were reached approximately 6 amplification cycles earlier compared to the commonly used FRET-based closed-tube PCR method. The suitability of the complementation probe method for different nucleic acid assay applications was studied. 1) A duplex complementation probe C. trachomatis PCR assay with a simple 10-minute urine sample preparation was developed to study suitability of the method for clinical diagnostics. The performance of the C. trachomatis assay was equal to the commercial C. trachomatis nucleic acid amplification assay containing more complex sample preparation based on DNA extraction. 2) A PCR assay for the detection of HLA-DQA1*05 allele, that is used to predict the risk of type 1 diabetes, was developed to study the performance of the method in genotyping. A simple blood sample preparation was used where the nucleic acids were released from dried blood sample punches using high temperature and alkaline reaction conditions. The complementation probe HLA-DQA1*05 PCR assay showed good genotyping performance correlating 100% with the routinely used heterogenous reference assay. 3) To study the suitability of the complementation probe method for direct measurement of the target organism, e.g., in the culture media, the complementation probes were applied to amplificationfree closed-tube bacteriophage quantification by measuring M13 bacteriophage ssDNA. A low picomolar bacteriophage concentration was detected in a rapid 20- minute assay. The assay provides a quick and reliable alternative to the commonly used and relatively unreliable UV-photometry and time-consuming culture based bacteriophage detection methods and indicates that the method could also be used for direct measurement of other micro-organisms. The complementation probe PCR method has a low background signal level leading to a high signal-to-background ratio and relatively sensitive nucleic acid detection. The method is compatible with simple sample preparation and it was shown to tolerate residues of urine, blood, bacteria and bacterial culture media. The common trend in nucleic acid diagnostics is to create easy-to-use assays suitable for rapid near patient analysis. The complementation probe PCR assays with a brief sample preparation should be relatively easy to automate and hence, would allow the development of highperformance nucleic acid amplification assays with a short overall assay time.

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Sosiaalinen media viittaa verkkopalveluihin, joiden toiminta perustuu käyttäjien ja käyttäjäyhteisöjen aktiiviseen sosiaaliseen vuorovaikutukseen. Matkapuhelimien kautta sosiaalinen media on liikkeellä ja pitää käyttäjät yhdessä ajasta ja paikasta riippumatta. Sosiaalisen median vetovoima perustuu siihen, että ihmiset jakavat tietoja, kuvia, videota ja ääntä sekä kommentoivat ja jatkavat viestiketjuja. Yhdessä ja liikkeellä -kirja tarkastelee sosiaalisen median uusinta ilmiötä, videoiden jakamista kännykkäpalvelun kautta. Mobiilit videot voivat kertoa jaetuista elämyksistä, konserteista, benji-hypyistä tai jazz-festivaalin vessajonosta. Liikkuva kuva kertoo usein enemmän kuin useampi valokuva, ja monen käyttäjän videokuva samasta tapahtumasta voi jo kertoa monipolvisen tarinan. Yhdessä ja liikkeellä esittelee Mobile Social Media -tutkimushankkeen tuloksia. Tavoitteena on ollut tutkia, millaiset tekijät tukevat mobiilin sosiaalisen median käyttöä ja miten sosiaalista mediaa hyödynnetään matkapuhelimilla, miten sosiaalisen median yhteisö voi muodostaa kännykkävideoista yhteisen tarinan, millaisia teknologisia haasteita mobiilin sosiaalisen median käytölle on. Tutkimushanke on CAT - Culture Art and Technology -verkoston yhteistyötä. Verkostoon kuuluvat Aalto-yliopiston, Tampereen teknillisen yliopiston ja Turun yliopiston Porin yksiköt.

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Tutkimuksessa analysoidaan yhden aikakauslehden aineettoman pääoman keskeisiä tekijöitä ja niiden dynamiikkaa laadullisilla menetelmillä. Aineisto perustuu haastattelumateriaaliin. Taustan muodostaa tietojohtamisen teoriakirjallisuus, jossa eri tietopääomatekijät jaotellaan suhde-, rakenne ja inhimillisiin tekijöihin. Aikakauslehdet, kuten koko mediatoimiala, ovat vahvassa muutostilanteessa ja aineettoman pääoman tekijöiden kautta ne voivat jatkossakin ylläpitää markkina-asemaansa. Tutkimuksen kohteena on hyvän markkina-aseman omaava aikakauslehti, jonka arvonmuodostusta ja aineettomia tekijöitä ja niiden välistä dynamiikkaa tutkimuksessa analysoidaan. Tutkimustulokset osoittavat, että tarkasteltavan aikakauslehden arvonmuodostus perustuu vahvaan aineettomien tekijöiden kombinaatioon, vahvaan organisaatiokulttuuriin ja aineettomien tekijöiden väliseen dynamiikkaan, mikä saadaan ulosmitattua printtituotteen muodossa kuluttajamarkkinoilla. Haasteena on näiden pääomatekijöiden hyödyntäminen yhtä tehokkaalla tavalla muuttuvan markkinatilanteen myötä.

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Long-term independent budget travel to countries far away has become increasingly common over the last few decades, and backpacking has now entered the tourism mainstream. Nowadays, backpackers are a very important segment of the global travel market. Backpacking is a type of tourism that involves a lot of information search activities. The Internet has become a major source of information as well as a platform for tourism business transactions. It allows travelers to gain information very effortlessly and to learn about tourist destinations and products directly from other travelers in the form of electronic word-of-mouth (eWOM). Social media has penetrated and changed the backpacker market, as now modern travelers can stay connected to people at home, read online recommendations, and organize and book their trips very independently. In order to create a wider understanding on modern-day backpackers and their information search and share behavior in the Web 2.0 era, this thesis examined contemporary backpackers and their use of social media as an information and communication platform. In order to achieve this goal, three sub-objectives were identified: 1. to describe contemporary backpacker tourism 2. to examine contemporary backpackers’ travel information search and share behavior 3. to explore the impacts of new information and communications technologies and Web 2.0 on backpacker tourism The empirical data was gathered with an online survey, thus the method of analysis was mainly quantitative, and a qualitative method was used for a brief analysis of open questions. The research included both descriptive and analytical approaches, as the goal was to describe modern-day backpackers, and to examine possible interdependencies between information search and share behavior and background variables. The interdependencies were tested for statistical significance with the help of five research hypotheses. The results suggested that backpackers no longer fall under the original backpacker definitions described some decades ago. Now, they are mainly short-term travelers, whose trips resemble more those of mainstream tourists. They use communication technologies very actively, and particularly social media. Traditional information sources, mainly guide books and recommendations from friends, are of great importance to them but also eWOM sources are widely used in travel decision making. The use of each source varies according to the stage of the trip. All in all, Web 2.0 and new ICTs have transformed the backpacker tourism industry in many ways. Although the experience has become less authentic in some travelers’ eyes, the backpacker culture is still recognizable.

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Social media is very current topic in today’s society and organisations. In the times of economic challenges, companies are looking for efficient ways to resource workforce. In addition, there is competition of competent workforce in employment markets. Employer image plays important role in recruitment as people seek to organisations they find interesting and have a reputation as a good employer. This study concerns the discussion on utilising social media in recruitment and employer image in a corporate enterprise. I will find solutions how to utilize social media in recruitment, in which channels and methods that can be done and what these actions require from a company doing social recruitment. I bring up the discussion and challenges that relate to starting to take social media into use in an organization overall and in recruitment. The qualitative material has been gathered with interviews of eighteen persons and the material available about the topic in the enterprise intranet. According to the study, social media is seen both as an opportunity to reach large amount of people quickly and cost-efficiently, but then again it brings news aspects for controlling the employer image and communication towards the audience. Taking social media into use as part of recruiters and managers daily work requires both finding the right channels and attention to the internal communication culture and resourcing. Social recruitment requires a strategy and a proper plan to be able to work in a company. There are several social media channels that enable to reach people, but they don’t do the social recruitment alone.