Cellular localisation of type XIII collagen, and its induced expression in human neoplasias and corneal diseases

Abstract Type XIII collagen belongs to the group of transmembrane collagens. In this thesis the plasma membrane localisation and function of type XIII collagen have been studied using cell biological methods. Type XIII collagen was found to reside in focal adhesions. It appeared in these structures at a very early stage of their assembly and disappeared from them concurrently with focal adhesion proteins talin and vinculin. Insect cells expressing type XIII collagen showed an enhanced adhesion to certain matrix components. These localisation and adhesion data suggested that the function of type XIII collagen is related to cell adhesion. Supporting this, in tissues type XIII collagen was found to localise to cell-matrix and cell-cell adhesion structures. Type XIII collagen was found to be partly present in cholesterol-enriched membrane microdomains. With other membrane proteins this localisation has been shown to be linked to ectodomain shedding. The connection between the membrane microdomain localisation and the ectodomain shedding of type XIII collagen was also characterised, and it was demonstrated that manipulation of the cellular cholesterol level affected the efficiency of the ectodomain shedding. Additionally, insights into intracellular shedding of type XIII collagen in the Golgi apparatus were obtained. The study of type XIII collagen expression in human cancers revealed that it was enhanced especially in the desmoplastic cancer stroma. Since the increased expression of type XIII collagen was detected during the dysplastic stages, type XIII collagen may be involved in the early pathogenesis of cancer. The result indicated that type XIII collagen is involved in the matrix remodelling. In support of this, the cell culture experiments showed that the soluble type XIII collagen ectodomain altered the vitronectin-rich matrix unfavourable for cell adhesion and spreading. This may enhance cancer metastasis. Type XIII collagen expression was also induced in the remodelled stroma of keratoconus and corneal wounds. Data suggested that myofibroblasts were responsible for the increased expression of type XIII collagen in these situations. Therefore both in cancer and in the corneal pathologies studied, type XIII collagen expression was induced by the activated stromal cells

Validation of a new measuring system for water turbidity field measurements

Turbidity is an essential parameter for describing water quality by direct and indirect impacts on fish, invertebrates and aquatic plants. Currently, environmental monitoring measurements are carried out with appropriate quality by accredited testing laboratories, but there is also a need for employing the third sector, i.e. citizens and voluntaries in environmental monitoring. A device called "Secchi3000" was developed as a low-cost and simple-to-operate tool so that water quality measurements can also be carried out by non-experts and citizens. The measurement using the new device is simple: The user fills the container with water and places the measurement structure in the container. The user takes a photograph with the camera on a mobile phone through a hole in the lid. The software sends the photograph to a server, which analyses the photographs automatically. Finally, the results are returned to the user's mobile phone and stored on a database for further analysis. In this study, the measuring system for turbidity measurements in natural waters was validated. Validation included an estimation of the limit of quantification, investigations of the influence of water colour and illumination conditions on turbidity measurement values and the estimation of measurement uncertainty. A comparison of turbidity results obtained with the new device and laboratory instrument in natural water samples was carried out, and turbidity values obtained with different mobile phones were compared. According to the validation results, the new device was appropriate for the measurement of turbidity lower than 7 FNU (Formazine Nephelometric Unit). An algorithm applied for present turbidity calculations is not fully suitable for higher turbidities. For potential routine use, this is not a major problem, since most Finnish natural waters have turbidities lower than 7 FNU. For official monitoring purposes, the limit of the quantification needs to be lower than presently achieved (1.7 FNU). Although the present configuration of the Secchi3000 device is not yet fully suitable for official environmental monitoring, it will already enable the involvement of the third sector in water quality monitoring, and in this way, citizens' observations could then serve at least as supplementary information for reporting and surveys

Partitioning of benthic biodiversity in boreal streams:contributions of spatial, inter-annual, and seasonal variability

Abstract 1. Studies on biodiversity patterns should optimally relate different scales of temporal community variability to spatial variability. Although temporal biodiversity variability is often negligible compared to spatial variation, it may still constitute a substantial source of overall community variability in stream ecosystems. Boreal streams exhibit seasonally recurring environmental periodicity, which can be expected to induce synchronous dynamics of abiotic variables among sites, and consequently, to produce spatial synchrony of deterministically controlled biological communities with higher intra- than inter-annual community variability. 2. We sampled benthic macroinvertebrates in 10 near-pristine boreal streams on three different seasons (spring, summer, autumn) across 4 consecutive years in northern Finland. We aimed to identify the relative contributions of spatial, inter-annual, and seasonal variability to overall benthic biodiversity; and relate variation in benthic invertebrate communities to key environmental factors, particularly in-stream habitat diversity. 3. Among-site spatial variability was clearly the most important source of variation for both species richness and community dissimilarity. Of the two temporal scales, inter-annual variability contributed more to variation in taxonomic richness and seasonal variability slightly more to variability in community composition. 4. Only inter-annual variation differed systematically from random expectation, indicating strong stability (low variability) of stream macroinvertebrate communities across years, with less variation at sites with higher substrate heterogeneity. Considering the distinct seasonality of the boreal stream environment, seasonal variability accounted for an unexpectedly low amount of total community variability. 5. Although differences between seasons were small, autumnal sampling is likely to be the least susceptible to climatic vagaries, thus providing the most consistent and predictable conditions for benthic sampling in boreal streams, particularly for bioassessment purposes. Exceptional climatic conditions are becoming more frequent in northern Europe, probably causing substantial and largely unpredictable changes in benthic community composition. As a result, the importance of temporal (relative to spatial) community variability may increase

Consumer adoption of future MyData-based preventive eHealth services:an acceptance model and survey study

Abstract Background: Constantly increasing health care costs have led countries and health care providers to the point where health care systems must be reinvented. Consequently, electronic health (eHealth) has recently received a great deal of attention in social sciences in the domain of Internet studies. However, only a fraction of these studies focuses on the acceptability of eHealth, making consumers’ subjective evaluation an understudied field. This study will address this gap by focusing on the acceptance of MyData-based preventive eHealth services from the consumer point of view. We are adopting the term "MyData", which according to a White Paper of the Finnish Ministry of Transport and Communication refers to "1) a new approach, a paradigm shift in personal data management and processing that seeks to transform the current organization centric system to a human centric system, 2) to personal data as a resource that the individual can access and control." Objective: The aim of this study was to investigate what factors influence consumers’ intentions to use a MyData-based preventive eHealth service before use. Methods: We applied a new adoption model combining Venkatesh’s unified theory of acceptance and use of technology 2 (UTAUT2) in a consumer context and three constructs from health behavior theories, namely threat appraisals, self-efficacy, and perceived barriers. To test the research model, we applied structural equation modeling (SEM) with Mplus software, version 7.4. A Web-based survey was administered. We collected 855 responses. Results: We first applied traditional SEM for the research model, which was not statistically significant. We then tested for possible heterogeneity in the data by running a mixture analysis. We found that heterogeneity was not the cause for the poor performance of the research model. Thus, we moved on to model-generating SEM and ended up with a statistically significant empirical model (root mean square error of approximation [RMSEA] 0.051, Tucker-Lewis index [TLI] 0.906, comparative fit index [CFI] 0.915, and standardized root mean square residual 0.062). According to our empirical model, the statistically significant drivers for behavioral intention were effort expectancy (beta=.191, P<.001), self-efficacy (beta=.449, P<.001), threat appraisals (beta=.416, P<.001), and perceived barriers (beta=−.212, P=.009). Conclusions: Our research highlighted the importance of health-related factors when it comes to eHealth technology adoption in the consumer context. Emphasis should especially be placed on efforts to increase consumers’ self-efficacy in eHealth technology use and in supporting healthy behavior

Increased mesenchymal podoplanin expression is associated with calcification in aortic valves

Abstract Background and aim of the study: Calcific aortic valve disease (CAVD) is a progressive disease starting from mild valvular sclerosis and progressing to severe aortic stenosis (AS) with calcified valves. The origin of the calcification is proposed to be mesenchymal cells which have differentiated towards an osteoblastic phenotype. Podoplanin is a glycoprotein expressed in the endothelium of lymphatic vessels and in osteoblasts and osteocytes, mesenchymal cells, as well as in many carcinomas and aortic atherosclerotic lesions. In CAVD, its expression has been evaluated only as a marker of the lymphatic vasculature. Materials and methods: We determined podoplanin expression in human aortic valves in four patient groups: control (C, n=7), aortic regurgitation (AR, n=8), aortic regurgitation and fibrosis (AR + f, n=15) and AS (n=49) by immunohistochemistry and quantitative real-time PCR (RT-PCR). Results: Immunohistochemically, podoplanin expression was significantly increased in AR + f and AS groups when compared with the control and AR groups and the level of expression positively correlated with the extent of calcification and vascularity. Podoplanin mRNA levels were 1.7-fold higher in the AS group as compared with the control group (P=.05). Podoplanin-positivity was present not only in lymphatic vessel endothelium but also in osteoblasts, osteocytes, chondrocytes, macrophages and extracellular matrix. The majority of the podoplanin-positivity was in spindle cells with a myofibroblastic phenotype, often associated with calcifications. Tricuspid valves had more calcification-associated podoplanin than bi/unicuspid valves (median 1.52 vs 1.16, P<.001). Conclusions: CAVD is characterized by an increased expression of podoplanin; this is associated with the differentiation of valvular interstitial cells into calcium-producing, myofibroblast-like cells. In addition, tricuspid valves express relatively more podoplanin than bi/unicuspid valves

Notch downregulation and extramedullary erythrocytosis in hypoxia-inducible factor prolyl 4-hydroxylase 2-deficient mice

Abstract Erythrocytosis is driven mainly by erythropoietin, which is regulated by hypoxia-inducible factor (HIF). Mutations in HIF prolyl 4-hydroxylase 2 (HIF-P4H-2) (PHD2/EGLN1), the major downregulator of HIFα subunits, are found in familiar erythrocytosis, and large-spectrum conditional inactivation of HIF-P4H-2 in mice leads to severe erythrocytosis. Although bone marrow is the primary site for erythropoiesis, spleen remains capable of extramedullary erythropoiesis. We studied HIF-P4H-2-deficient (Hif-p4h-2gt/gt) mice, which show slightly induced erythropoiesis upon aging despite nonincreased erythropoietin levels, and identified spleen as the site of extramedullary erythropoiesis. Splenic hematopoietic stem cells (HSCs) of these mice exhibited increased erythroid burst-forming unit (BFU-E) growth, and the mice were protected against anemia. HIF-1α and HIF-2α were stabilized in the spleens, while the Notch ligand genes Jag1, Jag2, and Dll1 and target Hes1 became downregulated upon aging HIF-2α dependently. Inhibition of Notch signaling in wild-type spleen HSCs phenocopied the increased BFU-E growth. HIFα stabilization can thus mediate non-erythropoietin-driven splenic erythropoiesis via altered Notch signaling

The prognostic and predictive roles of plasma C-reactive protein and PD-L1 in non-small cell lung cancer

Abstract Background: Anti-PD-(L)1 agents have revolutionized the treatment paradigms of non-small cell lung cancer (NSCLC), while predictive biomarkers are limited. It has been previously shown that systemic inflammation, indicated by elevated C-reactive protein (CRP) level, is associated with a poor prognosis in anti-PD-(L)1 treated. The aim of the study was to analyze the prognostic and predictive value of CRP in addition to traditional prognostic and predictive markers and tumor PD-L1 score. Methods: We identified all NSCLC patients (n = 329) who had undergone PD-L1 tumor proportion score (TPS) analysis at Oulu University Hospital 2015–22. CRP levels, treatment history, immune checkpoint inhibitor (ICI) therapy details, and survival were collected. The patients were categorized based on CRP levels (≤10 vs. >10) and PD-L1 TPS scores (<50 vs. ≥50). Results: In the whole cohort (n = 329), CRP level of ≤10 mg/L was associated with improved survival in univariate (HR 0.30, Cl 95% 0.22–0.41) and multivariate analyzes (HR 0.44, CI 95% 0.28–0.68). With ICI treated (n = 70), both CRP of ≤10 and PD-L1 TPS of ≥50 were associated with improved progression-free survival (PFS) in univariate (HR 0.51, CI 95% 0.27–0.96; HR 0.54, CI 95% 0.28–1.02) and multivariate (HR 0.48, CI 95% 0.26–0.90; HR 0.50, CI 95% 0.26–0.95) analyzes. The combination (PD-L1 TPS ≥50 and CRP >10) carried a high negative predictive value with a median PFS of 4.11 months (CI 95% 0.00–9.63), which was similar to patients with low PD-L1 (4.11 months, CI 95% 2.61–5.60). Conclusions: Adding plasma CRP levels to PD-L1 TPS significantly increased the predictive value of sole PD-L1. Furthermore, patients with high CRP beard little benefit from anti-PD-(L)1 therapies independent of PD-L1 score. The study highlights the combined evaluation of plasma CRP and PD-L1 TPS as a negative predictive marker for ICI therapies

Targeting collagen XVIII improves the efficiency of ErbB inhibitors in breast cancer models

Abstract The tumor extracellular matrix (ECM) critically regulates cancer progression and treatment response. Expression of the basement membrane component collagen XVIII (ColXVIII) is induced in solid tumors, but its involvement in tumorigenesis has remained elusive. We show here that ColXVIII was markedly upregulated in human breast cancer (BC) and was closely associated with a poor prognosis in high-grade BCs. We discovered a role for ColXVIII as a modulator of epidermal growth factor receptor tyrosine kinase (ErbB) signaling and show that it forms a complex with ErbB1 and -2 (also known as EGFR and human epidermal growth factor receptor 2 [HER2]) and α6-integrin to promote cancer cell proliferation in a pathway involving its N-terminal portion and the MAPK/ERK1/2 and PI3K/AKT cascades. Studies using Col18a1 mouse models crossed with the mouse mammary tumor virus–polyoma virus middle T antigen (MMTV-PyMT) mammary carcinogenesis model showed that ColXVIII promoted BC growth and metastasis in a tumor cell–autonomous manner. Moreover, the number of mammary cancer stem cells was significantly reduced in the MMTV-PyMT and human cell models upon ColXVIII inhibition. Finally, ablation of ColXVIII substantially improved the efficacy of ErbB-targeting therapies in both preclinical models. In summary, ColXVIII was found to sustain the stemness properties of BC cells and tumor progression and metastasis through ErbB signaling, suggesting that targeting ColXVIII in the tumor milieu may have important therapeutic potential

Novel human lymph node-derived matrix supports the adhesion of metastatic oral carcinoma cells

Abstract Background: 3D culture is increasingly used in cancer research, as it allows the growth of cells in an environment that mimics in vivo conditions. Metastases are the primary cause of morbidity and mortality in cancer patients, and solid tumour metastases are mostly located in lymph nodes. Currently, there are no techniques that model the pre-metastatic lymph node microenvironment in vitro. In this study, we prepared a novel extracellular matrix, Lymphogel, which is derived from lymph nodes, mimicking the tumour microenvironment (TME) of metastatic carcinoma cells. We tested the suitability of the new matrix in various functional experiments and compared the results with those obtained using existing matrices. Methods: We used both commercial and patient-derived primary and metastatic oral tongue squamous cell carcinoma (OTSCC) cell lines. We characterized the functional differences of these cells using three different matrices (human uterine leiomyoma-derived Myogel, human pre-metastatic neck lymph node-derived Lymphogel (h-LG), porcine normal neck lymph node-derived Lymphogel (p-LG) in proliferation, adhesion, migration and invasion assays. We also performed proteomic analyses to compare the different matrices in relation to their functional properties. Results: OTSCC cells exhibited different adhesion and invasion patterns depending on the matrix. Metastatic cell lines showed improved ability to adhere to h-LG, but the effects of the matrices on cell invasion fluctuated non-significantly between the cell lines. Proteomic analyses showed that the protein composition between matrices was highly variable; Myogel contained 618, p-LG 1823 and h-LG 1520 different proteins. The comparison of all three matrices revealed only 120 common proteins. Analysis of cellular pathways and processes associated with proteomes of each matrix revealed similarities of Myogel with h-LG but less with p-LG. Similarly, p-LG contained the least adhesion-related proteins compared with Myogel and h-LG. The highest number of unique adhesion-related proteins was present in h-LG. Conclusions: We demonstrated that human pre-metastatic neck lymph node-derived matrix is suitable for studying metastatic OTSCC cells. As a whole-protein extract, h-LG provides new opportunities for in vitro carcinoma cell culture experiments

Haitallisten vieraslajien hallinta ja tietoisuuden lisääminen (HAVINA) : loppuraportti

Liitteet: Liite 1. Tutkimusosapuolet ja yhteistyö (s. 26-27) Liite 2. HAVINA -hankkeeseen liittyvät julkaisut ja esitelmät (s. 28-31) Liite 3. HAVINA – Vieraslajiportaali 4 s. URN:NBN:fi-fe201702011419 (Luonnontieteellinen keskusmuseo – Luomus). Liite 4. Sisällöntuotto-ohje vieraslajiportaaliin. 10 s. URN:NBN:fi-fe201702011418 (Luomus) Liite 5. Haitallisten vieraiden kala-, rapu- ja nisäkäslajien leviäminen, tietoisuuden lisääminen ja hallinta (HAVINA osaraportti) 76 s. URN:ISBN:978-952-303-111-1 (RKTL) Liite 6. Sisävesien vieraslajit – vesikasvit. 15 s. URN:NBN:fi-fe201701311402 (SYKE vesikeskus) Liite 7. Selvitys meriteitse Suomeen kulkeutuvien vieraslajien saapumisreiteistä. 11 s. URN:NBN:fi-fe201701311401 (SYKE merikeskus) Liite 8. Ruohovartisten koristekasvien leviämiskartoitus ja -ennuste olemassa olevien havaintojen perusteella. 16 s. URN:NBN:fi-fe201701311399. (MTT, Kasvintuotannon tutkimus, Piikkiö & Jokioinen) Liite 9. Vieraslajien torjunta Lounais-Suomessa –hyvät käytännöt. 43 s. URN:ISBN:978-952-257-811-2 (Varsinais-Suomen Elykeskus) Liite 10. Kasvintuhoojien leviämisväylät ja kasvintuhoojien uhkaaman tuotannon arvo 76 s. (rinnakkaistallenne) (EVIRA & MTT, taloustutkimus) Liite 11. Ohjeistus vieraslajien torjunnan taloudelliseen arviointiin 33 s. URN:ISBN:978-952-487-528-8 (MTT, taloustutkimus & IEEP) Liite 12. Instruments for financing action on invasive alien species (IAS). 45 s. URN:NBN:fi-fe201701311400. (IEEP & MTT, taloustutkimus) Liite 13. Suomen luonnonsuojeluliiton tehtävät HAVINA-hankkeessa. 5 s. URN:NBN:fi-fe201702011420Sisällysluettelo: Loppuraportin tiivistelmä, s. 2-5 Hankkeen suositukset päättäjille, käytännön toimijoille ja suunnittelijoille sekä kansalaisille s. 6-12 1. Tutkimuksen tavoitteet, s. 14 2. Tutkimusmenetelmät ja aineisto, s. 14-16 3. Tutkimuksen tulokset ja johtopäätökset, s. 16-22. 4. Tulosten arviointi ja tietotarpeet, s. 23-25. 5. Yhteenveto suosituksista, s. 25-26.