Markkinointiviestintäsuunnitelma Helsingin Fillariosa Oy:lle

Opinnäytetyömme tehtiin toimeksiantona Fillariosa Oy:lle, jonka verkkokaupan sekä huoltotoiminnan markkinointiin ei oltu aiemmin kiinnitetty suurempaa huomiota. Opinnäytetyön tarkoituksena on laatia yritykselle käyttökelpoinen ja tarpeen mukaan muokattavissa oleva markkinointiviestintäsuunnitema vuodelle 2013 yrityksen rahalliset resurssit huomioiden. Tarkoituksena on myös kartoittaa yritykselle sopivia, kustannus-tehokkaita markkinointiviestinnän kanavia, kuten Googlen hakusanamainontaa ja Facebook-markkinointia. Työ toteuttettiin produktina. Koska huolto- ja verkkokauppatoiminta ovat vahvasti toisiaan tukevia, päädyttiin ne kummatkin ottamaan huomioon markkinointiviestintäsuunnitelmassa. Työn teoriaosuus avaa yrityksen toimintaympäristöä, markkinoinnin tavoitteita ja muo-toja sekä kilpailustrategiavalintoja. Lisäksi teoriassa avataan markkinointiviestinnän muotoja Nick Burcherin BOE-mallin kautta ja haastetaan perinteinen 4P-malli Sami Salmenkiven ja Niko Nymanin CREF-mallin kautta. Teoriaosuuden jälkeen työssä ava-taan produktia yrityksen esittelyllä, kuvaamalla markkinoinnin nykytilaa BOE-mallin kautta, sekä kuvaamalla markkinoita, joilla yritys toimii. Tämän jälkeen yritystä analy-soidaan SWOTin, ja kilpailija-analyysin keinoin ja markkinoinnin toimenpiteitä pohja-taan CREF-mallin mukaisesti. Teoriaan ja analyysiin pohjautuen yrityksen käyttöön on tuotettu markkinointisuunnitelma joka avataaan kuukausitasolla. Suhteellisen kapeilla markkinoilla Fillariosan tulee panostaa löydettävyyteen, kilpailijoista erottautumiseen ja pyrkiä luomaan pitkiä asiakassuhteita. Fillariosan tulee hahmottaa kohderyhmänsä ja ne markkinointiviestinnän kanavat, joilla nuo kohderyhmät parhaiten tavoitetaan. Hakukonemainonnalla ja Facebook mainonnalla sekä harrastajien aikakaus-lehtien ja tapahtumien kautta yritys pääsee kustannustehokkaasti näkyville medioissa, joista kohderyhmä tavoitetaan.Our thesis was comissioned by Fillariosa Oy, a small Finnish company which is run by an independent entrepreneur. Its webstore and bike service marketing have not been thought through earlier due to limited financial and time resources. The objective of the thesis was to create a useful and malleable marketing communications plan for the year 2013 taking the limited budget of a small business into consideration. The objective was also to scout suitable and cost efficent marketing channels for the company, ie. Google Search and Facebook marketing possibilities. The study and the marketing communications plan take both webstore and services into account as they were strongly affecting one another. The theoretical framework is based on the literature about marketing environment, marketing objectives and forms and the competitive strategy choices that a company has to make. Marketing communication channels were examined through Nick Burcher's BOE-model and the traditional 4P-model is being challenged with CREF-model made by Sami Salmenkivi and Niko Nyman. After the theoretical part of the thesis, the company and the market it works on is introduced, and the current stage of company's marketing communications is examined through BOE-model. The company is then analyzed with SWOT and competitor analysis. The CREF-model is used as the basis of the marketing communications plan. Based on the theoretical frameworks and company analysis, the marketing communications plan is then produced in a form of monthly calendar of marketing activities. As Fillariosa operates on rather narrow markets, it must focus on findability, differentiation from the core competitors and creating long term customer relationships in order to succeed. As a conclusion Fillariosa needs to comprehend its target groups and the marketing channels through which it can reach its target groups the most effectively. Through search engine and Facebook marketing, target group magazines and enthusiast events the company can reach widely and cost efficiently the enthusiastic target groups it wants to sell the products and services to

Metagenomic and metatranscriptomic analysis of the microbial community in Swiss-type Maasdam cheese during ripening

In Swiss-type cheeses, characteristic nut-like and sweet flavor develops during the cheese ripening due to the metabolic activities of cheese microbiota. Temperature changes during warm and cold room ripening, and duration of ripening can significantly change the gene expression of the cheese microbiota, which can affect the flavor formation. In this study, a metagenomic and metatranscriptomic analysis of Swiss-type Maasdam cheese was performed on samples obtained during ripening in the warm and cold rooms. We reconstructed four different bacterial genomes (Lactococcus lactis, Lactobacillus rhamnosus, Lactobacillus helveticus, and Propionibacterium freudenreichii subsp. shermanii strain JS) from the Maasdam cheese to near completeness. Based on the DNA and RNA mean coverage, Lc. lactis strongly dominated (similar to 80-90%) within the cheese microbial community. Genome annotation showed the potential for the presence of several flavor forming pathways in these species, such as production of methanethiol, free fatty acids, acetoin, diacetyl, acetate, ethanol, and propionate. Using the metatranscriptomic data, we showed that, with the exception of Lc. lactis, the central metabolism of the microbiota was downregulated during cold room ripening suggesting that fewer flavor compounds such as acetoin and propionate were produced. In contrast, Lc. lactis genes related to the central metabolism, including the vitamin biosynthesis and homolactic fermentation, were upregulated during cold room ripening.Peer reviewe

IRF2BP2 Mutation Is Associated with Increased STAT1 and STAT5 Activation in Two Family Members with Inflammatory Conditions and Lymphopenia

Interferon regulatory factor 2 binding protein 2 (IRF2BP2) is a transcriptional coregulator that has an important role in the regulation of the immune response. IRF2BP2 has been associated with the Janus kinase (JAK)—signal transducers and activators of transcription (STAT) pathway, but its exact role remains elusive. Here, we identified a novel clinical variant, IRF2BP2 c.625_665del, from two members of a family with inflammatory conditions and investigated the function of IRF2BP2 and c.625_665del mutation in JAK–STAT pathway activation and inflammatory signaling. The levels of constitutive and cytokine-induced phosphorylation of STATs and total STAT1 in peripheral blood monocytes, T cells, and B cells from the patients and four healthy controls were measured by flow cytometry. Inflammation-related gene expression was studied in peripheral blood mononuclear cells using direct digital detection of mRNA (NanoString). Finally, we studied the relationship between IRF2BP2 and STAT1 activation using a luciferase reporter system in a cell model. Our results show that patients having the IRF2BP2 c.625_665del mutation presented overexpression of STAT1 protein and increased constitutive activation of STAT1. In addition, interferon-induced JAK–STAT signaling was upregulated, and several interferon-inducible genes were overexpressed. Constitutive phosphorylation of STAT5 was also found to be upregulated in CD4+ T cells from the patients. Using a cell model, we show that IRF2BP2 was needed to attenuate STAT1 transcriptional activity and that IRF2BP2 c.625_665del mutation failed in this. We conclude that IRF2BP2 has an important role in suppressing immune responses elicited by STAT1 and STAT5 and suggest that aberrations in IRF2BP2 can lead to abnormal function of intrinsic immunity

Syöpä muuttaa solunulkoisten vesikkelien metabolista sormenjälkeä

Cancer alters cell metabolism. How these changes are manifested in the metabolite cargo of cancer-derived extracellular vesicles (EVs) remains poorly understood. To explore these changes, EVs from prostate, cutaneous T-cell lymphoma (CTCL), colon cancer cell lines, and control EVs from their noncancerous counterparts were isolated by differential ultracentrifugation and analyzed by nanoparticle tracking analysis (NTA), electron microscopy (EM), Western blotting, and liquid chromatography-mass spectrometry (LC-MS). Although minor differences between the cancerous and non-cancerous cell-derived EVs were observed by NTA and Western blotting, the largest differences were detected in their metabolite cargo. Compared to EVs from noncancerous cells, cancer EVs contained elevated levels of soluble metabolites, e.g., amino acids and B vitamins. Two metabolites, proline and succinate, were elevated in the EV samples of all three cancer types. In addition, folate and creatinine were elevated in the EVs from prostate and CTCL cancer cell lines. In conclusion, we present the first evidence in vitro that the altered metabolism of different cancer cells is reflected in common metabolite changes in their EVs. These results warrant further studies on the significance and usability of this metabolic fingerprint in cancer.Peer reviewe

IRF2BP2 Mutation Is Associated with Increased STAT1 and STAT5 Activation in Two Family Members with Inflammatory Conditions and Lymphopenia

Interferon regulatory factor 2 binding protein 2 (IRF2BP2) is a transcriptional coregulator that has an important role in the regulation of the immune response. IRF2BP2 has been associated with the Janus kinase (JAK)—signal transducers and activators of transcription (STAT) pathway, but its exact role remains elusive. Here, we identified a novel clinical variant, IRF2BP2 c.625_665del, from two members of a family with inflammatory conditions and investigated the function of IRF2BP2 and c.625_665del mutation in JAK–STAT pathway activation and inflammatory signaling. The levels of constitutive and cytokine-induced phosphorylation of STATs and total STAT1 in peripheral blood monocytes, T cells, and B cells from the patients and four healthy controls were measured by flow cytometry. Inflammation-related gene expression was studied in peripheral blood mononuclear cells using direct digital detection of mRNA (NanoString). Finally, we studied the relationship between IRF2BP2 and STAT1 activation using a luciferase reporter system in a cell model. Our results show that patients having the IRF2BP2 c.625_665del mutation presented overexpression of STAT1 protein and increased constitutive activation of STAT1. In addition, interferon-induced JAK–STAT signaling was upregulated, and several interferon-inducible genes were overexpressed. Constitutive phosphorylation of STAT5 was also found to be upregulated in CD4+ T cells from the patients. Using a cell model, we show that IRF2BP2 was needed to attenuate STAT1 transcriptional activity and that IRF2BP2 c.625_665del mutation failed in this. We conclude that IRF2BP2 has an important role in suppressing immune responses elicited by STAT1 and STAT5 and suggest that aberrations in IRF2BP2 can lead to abnormal function of intrinsic immunity

Serum Inhibin-A and PAPP-A2 in the prediction of pre-eclampsia during the first and second trimesters in high-risk women

Objectives: Maternal serum inhibin-A , pregnancy associated plasma protein-A (PAPP-A) and PAPP-A2 together with placental growth factor (PlGF), maternal risk factors and uterine arter y pulsatility inde x (UtA PI) were analysed to study thei r ability to predict pre-eclampsia (PE). Study design: Serial serum samples for the nested case-control study were collected prospectively at 12-14, 18-20 and 26-28 weeks of gestation from 11 women who later developed early-onset PE (EO PE , diagnosis < 34 + 0 weeks of gestation), 34 women who developed late-onset PE (LO PE , diagnosis 2 34 + 0 weeks) and 89 controls. Main outcome measures: Gestational age-adjusted multiples of the median (MoM) values were calculated for biomarker concentrations. Multivariate regression analyses were performed to combine first trimester bio-markers, previously reported results on PlGF, maternal risk factors and UtA PI. Area under cu r v e (AUC) values and 95% confidence intervals (CIs) for the prediction of PE and its subtypes were calculated . Results: A high first trimester inhibin-A predicted PE (AUC 0.618, 95%CI, 0.513-0.724), whereas PAPP-A and PlGF predicted only EO PE (0.701, 0.562-0.840 and 0.798, 0.686-0.909, respectively). At 26-28 weeks PAPP-A2 and inhibin-A predicted a l l PE subtypes. In the multivariate setting inhibin-A combined with maternal pre-pregnancy body mass index, prior PE and mean UtA PI predicted PE (0.811,0.726-0.896) and LO PE (0.824, 0.733-0.914). Conclusions: At first trimester inhibin-A show potential ability to predict not only EO PE but also LO PE whereas PlGF and PAPP-A predict only EO PE. At late second trimester inhibin-A and PAPP-A2 might be usef u l for short-term prediction of PE.Peer reviewe

Metabolic signature of extracellular vesicles depends on the cell culture conditions

One of the greatest bottlenecks in extracellular vesicle (EV) research is the production of sufficient material in a consistent and effective way using in vitro cell models. Although the production of EVs in bioreactors maximizes EV yield in comparison to conventional cell cultures, the impact of their cell growth conditions on EVs has not yet been established. In this study, we grew two prostate cancer cell lines, PC-3 and VCaP, in conventional cell culture dishes and in two-chamber bioreactors to elucidate how the growth environment affects the EV characteristics. Specifically, we wanted to investigate the growth condition-dependent differences by non-targeted metabolite profiling using liquid chromatography-mass spectrometry (LC-MS) analysis. EVs were also characterized by their morphology, size distribution, and EV protein marker expression, and the EV yields were quantified by NTA. The use of bioreactor increased the EV yield >100 times compared to the conventional cell culture system. Regarding morphology, size distribution and surface markers, only minor differences were observed between the bioreactor-derived EVs (BR-EVs) and the EVs obtained from cells grown in conventional cell cultures (C-EVs). In contrast, metabolomic analysis revealed statistically significant differences in both polar and non-polar metabolites when the BR-EVs were compared to the C-EVs. The results show that the growth conditions markedly affected the EV metabolite profiles and that metabolomics was a sensitive tool to study molecular differences of EVs. We conclude that the cell culture conditions of EV production should be standardized and carefully detailed in publications and care should be taken when EVs from different production platforms are compared with each other for systemic effects.Peer reviewe

gapFinisher: A reliable gap filling pipeline for SSPACE-LongRead scaffolder output

Unknown sequences, or gaps, are present in many published genomes across public databases. Gap filling is an important finishing step in de novo genome assembly, especially in large genomes. The gap filling problem is nontrivial and while there are many computational tools partially solving the problem, several have shortcomings as to the reliability and correctness of the output, i.e. the gap filled draft genome. SSPACE-LongRead is a scaffolding tool that utilizes long reads from multiple third-generation sequencing platforms in finding links between contigs and combining them. The long reads potentially contain sequence information to fill the gaps created in the scaffolding, but SSPACE-LongRead currently lacks this functionality. We present an automated pipeline called gapFinisher to process SSPACE-LongRead output to fill gaps after the scaffolding. gapFinisher is based on the controlled use of a previously published gap filling tool FGAP and works on all standard Linux/UNIX command lines. We compare the performance of gapFinisher against two other published gap filling tools PBJelly and GMcloser. We conclude that gapFinisher can fill gaps in draft genomes quickly and reliably. In addition, the serial design of gapFinisher makes it scale well from prokaryote genomes to larger genomes with no increase in the computational footprint.Peer reviewe

Seudullista kansalaisosallistumista jäljittämässä tiedon yhteistuottamisen keinoin

This article analyses an ongoing process of knowledge co-production concerning the role of citizens and participation in strategic city-regional planning, and discusses the theoretical and practical knowledge that results from the process. In knowledge co-production involving urban planning experts and researchers, three things stand out as key to successful outcomes: an ongoing dialogue between scientific and practical understanding, knowledge production as an accumulative process, and constructive criticism as a dynamism that pushes the frontiers of practical understanding. Based on our initial empirical findings about citizenship and participation on the city-regional level, we suggest that the role of broad value- and issue-based deliberation concerning the long-term aims of urban environment could be a fruitful starting point for discussions between citizens, experts and politicians within emerging city-regions.Peer reviewe

Genome description of Phlebia radiata 79 with comparative genomics analysis on lignocellulose decomposition machinery of phlebioid fungi

Background The white rot fungus Phlebia radiata, a type species of the genus Phlebia, is an efficient decomposer of plant cell wall polysaccharides, modifier of softwood and hardwood lignin, and is able to produce ethanol from various waste lignocellulose substrates. Thus, P. radiata is a promising organism for biotechnological applications aiming at sustainable utilization of plant biomass. Here we report the genome sequence of P. radiata isolate 79 originally isolated from decayed alder wood in South Finland. To better understand the evolution of wood decay mechanisms in this fungus and the Polyporales phlebioid clade, gene content and clustering of genes encoding specific carbohydrate-active enzymes (CAZymes) in seven closely related fungal species was investigated. In addition, other genes encoding proteins reflecting the fungal lifestyle including peptidases, transporters, small secreted proteins and genes involved in secondary metabolism were identified in the genome assembly of P. radiata. Results The PACBio sequenced nuclear genome of P. radiata was assembled to 93 contigs with 72X sequencing coverage and annotated, revealing a dense genome of 40.4 Mbp with approximately 14 082 predicted protein-coding genes. According to functional annotation, the genome harbors 209 glycoside hydrolase, 27 carbohydrate esterase, 8 polysaccharide lyase, and over 70 auxiliary redox enzyme-encoding genes. Comparisons with the genomes of other phlebioid fungi revealed shared and specific properties among the species with seemingly similar saprobic wood-decay lifestyles. Clustering of especially GH10 and AA9 enzyme-encoding genes according to genomic localization was discovered to be conserved among the phlebioid species. In P. radiata genome, a rich repertoire of genes involved in the production of secondary metabolites was recognized. In addition, 49 genes encoding predicted ABC proteins were identified in P. radiata genome together with 336 genes encoding peptidases, and 430 genes encoding small secreted proteins. Conclusions The genome assembly of P. radiata contains wide array of carbohydrate polymer attacking CAZyme and oxidoreductase genes in a composition identifiable for phlebioid white rot lifestyle in wood decomposition, and may thus serve as reference for further studies. Comparative genomics also contributed to enlightening fungal decay mechanisms in conversion and cycling of recalcitrant organic carbon in the forest ecosystems.Peer reviewe