Browsing by department "Biovetenskapliga institutionen"
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(2012)In my master's thesis I have studied the characteristics of pedagogically well-designed web-based learning environment, where the contents are presented according to the principles of biology didactics. I have created a website called Virtuaalimetsä (Virtual Forest). Virtuaalimetsä is a web-based learning environment of Finnish forests for the pupils of classes 5 and 6 in primary school. Virtuaalimetsä is made to support both teaching and learning of biology. It offers lots of information and a variety of tools to develop thinking skills. The information of Virtuaalimetsä has its basis in biological research, but the academic nature of this information has been modified to be understandable to children. In this modification the didactics of biology and natural sciences has had a great role. The contents of Virtuaalimetsä fulfill the requirements of National Core Curriculum for Basic Education. With the information from research of blended learning Virtuaalimetsä website has become an effective learning environment. In planning and creating Virtuaalimetsä I had four objectives: 1. Objectives concerning blended learning: to create a good web-based learning environment 2. Objectives concerning learning materials: to increase learners' knowledge and develop their thinking skills, and to create proper tools to achieve this 3. Objectives concerning teaching: to support traditional teaching with a meaningful web-based learning environment 4. Objectives concerning environmental education: to awake a sense of environmental responsibility especially in relation to forests. To achieve these objectives I have studied research about blended learning and didactics of biology. Characteristics of a good web-based learning environment appears to be a proper pedagogical goal, clear research-based information, tools to develop thinking skills, information structuring and intense contemplation, as also logical and pedagogically operational website with good navigation tools. As the leading rules of teaching biology can be mentioned emphasizing the wholeness of nature, system thinking, proper use of concepts, holistic examination of ecological phenomena, using children's previous ideas as a basis for learning and developing observation skills. Virtuaalimetsä has six parts: Metsäkartta (Forest Map), Metsäpolku (Forest Path), Metsäsanasto (Forest Vocabulary), Testaa taitosi (Test your skills), Metsän kasvit (Forest Plants) and Opettajalle (For teacher). This division has been made consistently with the learning tools, and most parts offer a specific way to enhance learning. Virtuaalimetsä is further divided into sub-parts according to different themes: Metsä elinympäristönä (Forest as a habitat), Metsätyypit (Forest types), Metsän kerrokset (Forest layers) and Metsäluonnon monimuotoisuus (Biodiversity in forests). Forest Map concentrates on mind maps and concept maps. These help children to create ideas of wholes and concepts. Concept maps enhance active and meaningful learning and develop learners' meta-cognitive skills. Forest Path has the biological information of Virtuaalimetsä. The information is so presented that learners can get a good picture of forest nature as a whole and as a systemic structure. Information is presented at a concrete level, and new information is continuously connected to the context of forest. Thus the information is kept together, and no single detail is left detached. Many biological concepts are presented, and they help to compose a meaningful image of forests and of the nature of biological phenomena. In Virtuaalimetsä forest environment also gives a familiar ground to examine matter cycle, energy flow and population dynamics. These phenomena represent different kinds of interaction patterns and causal structures in nature, and that is why they improve development of biological thinking and understanding. To awake environmentally responsible attitudes Virtuaalimetsä offers a great deal of necessary information to contemplate environmental problems. Test you skills is a place for exercises. Exercises help in repetition, they give a possibility to test skills and most importantly they activate learned information. Many of the exercises are describing, comparing and classifying exercises, which help to understand the concepts. Exercises that involve explaining help to organize and analyze information. Forest Plants -part has a link to Helsinki University's Pinkka -learning environment. In Pinkka there is a section made especially for Virtuaalimetsä with 30 forest plant species. Plant identification has its own part in Virtuaalimetsä, because knowledge about species has a crucial role in understanding nature and ecosystems. In Forest vocabulary all the concepts presented in Virtuaalimetsä are explained in alphabetical order. In For teachers the idea of Virtuaalimetsä is explained in nutshell. There are also tips for teachers of how to use Virtuaalimetsä in teaching, and how to use information and cognitive tools for example in teamwork, whole class discussions or outside the classroom. The structure of Virtuaalimetsä website has been made into a logical and easily navigated whole by dividing the site according to the different cognitive tools and forest themes. Links are clearly presented, and studying is guided by explaining the contents of each part in advance and by giving tips of how to move in the site. There are also left many possibilities for learners to plan their own course of study. Multiple presentations are present: the information is presented both as pictures and text, and to minimize cognitive load all the information of one subject is presented on the same page.
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(2016)Biotopes and biodiversity within the Metropolitan Area of Helsinki (cities of Helsinki, Espoo, Vantaa and Kauniainen) were prioritized with a spatial conservation prioritization software called Zonation. The aim of the thesis was to examine how different, largely urban or semi-urban, areas supported biodiversity and thus, indirectly, ecosystem services provisioning. Furthermore, the effects of the new City of Helsinki Strategic Plan 2050 on the prioritization were examined. The analyses were based on expert elicitations, in which different urban biotopes were evaluated in terms of how well they supported species richness and the occurrence of specialist species of 8 taxonomic groups. Following the biotope classification, an urban biotope map was compiled from various GIS sources. Based on the expert answers, these biotope maps were then converted to suitability maps for each of the eight taxonomic groups. The urban biotope map, as well as the suitability maps, were made according to two land-use versions: current land-use in the research area, and a scenario describing the actual City of Helsinki Strategic Plan 2050. Separate Zonation prioritizations were then made for both scenarios. Urban biotopes a-priori evaluated as important for biodiversity, such as lakes and their shores, ruderal areas, dry meadows, manor yards, shoreline meadows and herb-rich forests, became emphasized in Zonation prioritizations as well, whereas the currently heavily built areas received lowest conservation priority in the analysis. Thus, according to this study, densification of the current residential and built-up areas does not threaten the urban biodiversity or ecosystem services of the Metropolitan Area of Helsinki. According to the present analysis, the new Helsinki Strategic Plan would have some relatively small effects on biodiversity in the research area. While the plan proposes development in some top-value biodiversity areas, the estimated effects of the plan were on average only a 3.3% loss of biotopes across taxonomic groups. Nevertheless, the plans of Helsinki could influence biodiversity priority areas in neighboring cities, and I recommend that the broader Metropolitan Area should be treated as a whole from the perspective of urban biodiversity. Rather than preserving sets of defined species, multi-functionality and ecosystem services provided by urban biodiversity should be emphasized when planning for urban nature conservation. As biodiversity ultimately underlies and supports both of these objectives, it should be central in the urban planning as well. Spatial conservation prioritization can well be used for ecologically sustainable urban planning, as long as the quality of the input data is taken care of and different uncertainties and limitations of the analyses are recognized in interpretation.
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Phytoplankton biomass and its relation to phosphorus loading and thermal conditions in Lake Pääjärvi (2015)The purpose of this thesis was to explore the relationships between phytoplankton biomass in lake Pääjärvi and environmental factors. These factors were divided into two categories: external phosphorus loading (total phosphorus, PO4 and organic phosphorus) and thermal conditions (Schmidt stability, heat volume and parent thermocline depth). The data sets at my disposal spanned the years 1994-2010 and contained phytoplankton observations, measurements of lake water temperature and measurements of phosphorus loading from the main rivers and streams flowing into lake Pääjärvi. Heat volume and stability were calculated based on the provided temperature data using a macro for Microsoft Office. The same data were used to calculate parent thermocline depth using LakeAnalyzer. The main statistical analysis performed on these data sets was Redundancy Analysis (RDA) in Canoco for Windows. The data were divided into three different periods: June, July-August and September-October. Each period was examined separately. In addition to total phytoplankton biomass, phytoplankton main groups and selected diatom and flagellate taxa were also examined in relation to the environmental factors. Heat volume proved statistically significant in June in the analysis of total biomass and the main groups and in September-October in the analysis of selected diatom taxa. A statistically significant relationship between PO4 loading and phytoplankton biomass was only detected for Peridinium sp, Rhodomonas lacustris and Mallomonas caudata for July-August. The lack of significance for phosphorus loading in the other contexts may in part be attributed to the distances between the phytoplankton sampling location and the loading measurement sites. The loading would have to drift some time in the lake to traverse these distances before it would come into contact with the measured phytoplankton. During that time, the loading would be susceptible to variations in weather or hydrological conditions as well as susceptible to uptake by other organisms. This may be the reason, or a contributing reason, why phosphorus loading was not significant for most of the contexts explored. However, since phosphorus loading was significant in at least one context, it clearly exerts some influence on the phytoplankton in Pääjärvi. The greater significance for heat volume could be attributed to the cumulative nature of the variable as well as the lack of spatial distance between the temperature sampling distance and phytoplankton sampling location.
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(2010)Biologisesti aktiivisilla oligosakkarideilla on vaikutuksia kasvin kasvuun ja kehittymiseen. Tietyn tyyppiset oligosakkaridit voivat myös indusoida puolustusreaktion valikoivasti oligosakkaridista riippuen. Useat biologisesti aktiiviset oligosakkaridit on löydetty kasvien soluseinää keinotekoisesti hajottamalla. Pro gradu -tutkielman tarkoituksena oli karakterisoida kuusen (Picea abies) solukkolinjan A3/85 solususpensiokasvatukseen erittämiä oligosakkarideja. Kuusisolukko A3/85 on otollinen kandidaatti tutkimukseen, sillä sen on todettu erittävän solunulkoista ligniiniä suspensioliuokseen. Oligosakkarideja karakterisoitiin yhden ja neljän vuorokauden kasvatuksista. Alustan sokeripitoisuutta alennettiin neljän vuorokauden kasvatuksissa karakterisoinnin helpottamiseksi. Oligosakkaridien pitoisuudet voivat olla hyvin alhaisia, joten solujen tuottamia yhdisteitä seurattiin myös radioaktiivisen D-[U-14C]-glukoosin avulla. Kasvien soluseinässä yleiset glukuronihappo, galakturonihappo, ksyloosi, arabinoosi ja apioosi valmistetaan glukoosi-6-fosfaatista, joko myo-inositolihapetusreitin tai sokerihapetusreitin kautta. Lisäksi tutkittiin, muuttuuko radioaktiivisen leiman jakautuminen näytteissä, kun kasvatusliuoksessa on tai ei ole myo-inositolia. Kasvatusliuos fraktioitiin geelisuodatuskromatografialla. Fraktioiden sisältämät yhdisteet eroteltiin paperikromatografialla ja värjättiin hopeanitraatilla, aniliinivetyftalaatilla tai ninhydriinillä. Hopeanitraatti on hyödyllinen monosakkaridien, oligosakkaridien ja alditolien värjäyksessä. Radioaktiivisuuden kertymistä yhdisteisiin seurattiin nestetuikelaskimella ja autoradiografialla. Paperikromatografialla erotelluista yhdisteistä valittiin mielenkiintoiseksi koetut yhdisteet, jotka eristettiin preparatiivisella paperikromatografialla. Eristetyille yhdisteille tehtiin happohydrolysointi, borohydridikäsittely tai entsymaattinen Driselaasi -käsittely. Happohydrolysointi avaa sokeriyksiköiden väliset glykosidiset sidokset. Natriumborohydridipelkistys muuttaa oligosakkaridiketjun pelkistävän sokerin sokerialkoholiksi ja Driselaasi -käsittely avaa isoprimeveroosin Xyl-α-(1 –> 6)-Glc -sidosta lukuun ottamatta muut glykosidiset sidokset. 14C-leima on jakautunut myo-inositolin kanssa kasvatetun näytteen fraktioinnissa vahvemmin suurimolekyylisiin yhdisteisiin, kun taas ilman myo-inositolia kasvatetussa näytteessä suurin aktiivisuus D-[U-14C]-glukoosin jälkeen on trisakkaridien alueella. Suspensioliuoksista analysoitiin useita oligosakkarideja polymerisaatioasteella 1-4. Analysoiduista yhdisteistä kolme sisälsivät ksyloosia, jota solut voivat syntetoida joko myo-inositolin hapetusreitin tai glukoosin hapetusreitin kautta. Myo-inositolin puuttuminen alustasta lisäsi näiden leimattujen yhdisteiden pitoisuutta. Alustan myo-inositoli ei ole radioaktiivista, joten myo-inositolihapetusreitin kautta valmistetut monosakkaridit eivät näy autoradiografiassa. Vaikuttaisi siis siltä, että myo-inositolihapetusreitti on aktiivinen ainakin, jos solukolle tarjotaan myo-inositolia. Lisätty myo-inositoli vähentää sokerihapetusreitin aktiivisuutta. Työn aikana onnistuttiin eristämään ja osittain tunnistamaan useita kuusen suspensioliuoksen yhdisteitä. Myo-inositolihapetusreitti todettiin aktiiviseksi solukkokasvatuksessa, kun ravintoalustassa on myo-inositolia.
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(2012)Valtaosa maapallon eliölajeista odottaa yhä tieteellistä kuvausta. Perinteisten menetelmien soveltaminen miljoonien nimeämättömien lajien kuvaamiseen on kuitenkin hidasta ja edellyttää enemmän taksonomista työvoimaa, kuin mitä todellisuudessa lienee koskaan saatavilla. Uutena ongelmana morfologisin menetelmin kuvattujen lajien sisältä on yhä enenevässä määrin paljastunut jopa useita piileviä eli kryptisiä lajeja, jotka eroavat merkittävästi muilta kuin ulkoisilta ominaisuuksiltaan. Ratkaisuksi näihin haasteisiin on ehdotettu DNA-taksonomiaa; lyhyiden, vakioitujen DNA-sekvenssien käyttöä uusien lajien tunnistamiseksi ja kryptisten lajien erottamiseksi. Tähän tarkoitukseen soveltuvaksi alueeksi eläimille on osoittautunut osa sytokromi c oksidaasi I (COI) -geeniä. Tutkielmassani tarkastelen DNA-taksonomian soveltuvuutta vastikään kuvatun, ulkoisin tuntomerkein vaikeasti eroteltavan lajiparin Aphodius fimetarius (pihalantiainen) ja Aphodius pedellus (salalantiainen) lajinmääritykseen. Kysyn: Onko näiden kromosomituntomerkein erotettujen lajien tunnistaminen mahdollista COI -geenin sekvenssierojen avulla? Voiko lajit tunnistaa aiemmin esitetyin ulkoisin tuntomerkein? Mikä on tutkimuslajien maailmanlaajuinen levinneisyys? Kumpi lajeista esiintyy Suomessa, ja millainen on tämän lajin geneettinen populaatiorakenne? Selvittääkseni tutkimieni lantakuoriaisyksilöiden lajin käytin kahta geneettistä menetelmää: kromosomipreparaatteja ja COI-geenin sekvenssejä, sekä vähäisiin ulkoisiin tuntomerkkeihin perustuvaa määrityskaavaa. Selvitykseni salalantiaisen esiintymisestä Suomessa perustui kahteen koko maan kattavaan lantakuoriaisaineistoon, sekä itse Etelä- ja Länsi-Suomesta keräämääni aineistoon. Käytössäni oli myös museonäytteitä maailmalta, sekä GenBankissa julkaistuja COIsekvenssejä. COI-sekvensseissä havaitsemani erot jakoivat aineiston kahdeksi toisistaan merkittävästi eroavaksi lajiksi: lajien välillä oli 8,1 % ero emäsjärjestyksessä, mutta lajien sisältä löytyi vain vähän muuntelua (0,2–0,7 %). Sekvenssituntomerkit erottelivat salalantiaisen ja pihalantiaisen samoiksi lajeiksi kuin aiemmin kuvatut kromosomituntomerkit. Määrityskaava sijoitti yli puolet yksilöistä samoihin ryhmiin kuin sekvenssitkin, mutta monilla yksilöillä oli kummankin lajin tuntomerkkejä. Kehittämäni DNA-lajitunnisteet osoittivat, että Suomessa esiintyy tutkituista lajeista vain salalantiainen, ei pihalantiainen. COI-tuntomerkein tunnistin salalantiaisen ensimmäisenä täysin varmana havaintona myös Pohjois-Amerikasta sekä Nepalista. Suomessa tavatuista salalantiaisen 27 COI-haplotyypistä yksi on yleinen ja laajalle levinnyt, muut harvinaisempia jalevinneisyydeltään paikallisia. Suomalaisen haplotyyppiverkoston tähtimäinen muoto viittaa siihen, että salalantiainen olisi vastikään nopeasti levittäytynyt uusille alueille. Kokonaisuudessaan tulokset osoittivat DNA-lajitunnisteet tarkaksi työkaluksi piha- ja salalantiaisen erottelemiseksi. DNA-tuntomerkkien toimivuus perustuu siihen, että lajien välinen sekvenssivaihtelu on merkittävästi lajinsisäistä vaihtelua runsaampaa. Soveltamalla uutta tunnistustyökalua laajaan aineistoon osoitin yhden maamme yleisimmän kovakuoriaisen kuuluvan eri lajiin kuin aiemmin on oletettu, ja tämän lajin olevan maailmalla laajalle levinnyt. Ilman COIsekvenssien tarjoamaa erottelukykyä morfologiset tuntomerkit eivät olisi riittäneet asian selvittämiseen. Tulevaisuudessa olisi kiinnostavaa solveltaa kehittämiäni tuntomerkkejä näiden lajien maailmanlaajuisen levinneisyyden entistä tarkempaan selvittämiseen. Vertailu kotimaiseen museoaineistoon voisi valaista, onko pihalantiainen kuitenkin joskus asuttanut maatamme ja kenties joutunut väistymään nopeasti levittäytyvän sisarlajinsa tieltä. Tähän lajitunnisteet tarjoavat nopean ja varman keinon.
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(2015)Ihmistoiminta muuttaa elinympäristöjä maailmanlaajuisesti. Vaikutukset näkyvät niin yksittäisten lajien populaatioissa, lajien välisissä vuorovaikutuksissa kuin kokonaisten ekosysteemien toiminnassakin. Monimutkaisuutensa ja vaikean havainnoitavuutensa takia lajien väliset vuorovaikutukset ja erityisesti niiden yhteys ekosysteemien toimintaan tunnetaan osin huonosti. Ympäristömuutoksista ilmaston lämpeneminen vaikuttaa erityisen voimakkaasti arktisella alueella. Muuttuva ja yksinkertainen arktinen eliöyhteisö tarjoaakin loistavan malliekosysteemin lajien välisten vuorovaikutusten havainnoimiselle ja niiden rakenteen mallintamiselle. Tässä tutkielmassa selvitän pölyttäjäyhteisön rakennetta ja toimintaa yhden kasvilajin, lapinvuokon Dryas octopetala × integrifolia, pölytyksessä. Lapinvuokko on runsain kukkakasvi monilla pohjoisilla alueilla, ja sitä voidaankin pitää avainlajina paikallisissa pölytysyhteisöissä. Liittääkseni pölyttäjäyhteisön rakenteen lapinvuokon siementuotannon onnistumiseen havainnoin lapinvuokon pölyttäjäyhteisön vaihtelua tilassa ja ajassa. Tähän käytin kukanmuotoisia liimapahvista valmistettuja pyydyksiä. Tein liimakukkapyyntejä 21 koealalla Zackenbergin laaksossa Grönlannissa. Tutkin koealoilla lisäksi lapinvuokon siementuotantoa. Tutkimuksessani havaitsin, että lapinvuokon kukinnan ajoittumisessa on suurta alueellista vaihtelua. Ensimmäiset kukat puhkeavat heti pälvien ilmestyessä ja viimeisten yksilöiden kukinta alkaa vasta juuri ennen ensilumia. Myös lapinvuokon siementuotannon onnistuminen vaihtelee paljon (12,7 - 52,7 % kukista) alueellisesti. Alueellinen vaihtelu siementuotannon onnistumisessa johtuu osittain ympäristöolosuhteista, mutta myös pölyttäjäyhteisöllä on selvä vaikutuksensa. Lapinvuokolla tapahtuu itse- ja tuulipölytystä (8,6 % onnistuneesta siementuotannosta), mutta pölyttäjien läsnä ollessa siementuotanto on moninkertainen (33,7 %). Tutkimusalueellani Zackenbergissa maailmanlaajuisesti tärkeimmät pölyttäjät, mesipistiäiset, puuttuvat lähes tyystin. Runsaimpien pölyttäjien paikan ovat vallanneet kärpäset erityisen runsaslukuisten sukaskärpästen johdolla. Havaintojeni perusteella sukaskärpästen runsaus selittääkin parhaiten lapinvuokon siementuotannon onnistumista. Kesän edetessä havaitsin pölyttäjäyhteisön kasvavan huomattavasti sekä yksilö- että lajimäärältään. Pölyttäjäyhteisön runsastuminen kasvukauden edetessä antaa olettaa, että lapinvuokon pölytys voi parantua ainakin lyhyellä aikavälillä. Ilmaston lämmetessä pölyttäjien esiintymisen on havaittu liukuvan aikaisemmaksi kasvien kukintaa nopeammin. Näin ollen aikaisin kukkivat kasvit kohtaavat yhä enemmän pölyttäjiä ja myöhäisimmätkin kasvit onnistuvat tuottamaan siemeniä ennen syksyä, kun niiden kukinta siirtyy absoluuttisesti aikaisemmaksi. Toisaalta sukaskärpästen määrän on havaittu vähenevän ilmaston lämmetessä. Tutkimukseni pohjalta sukaskärpäset ovat lapinvuokon tärkeimpiä pölyttäjiä, ja jos niiden väheneminen jatkuu, on lapinvuokon pölytys Zackenbergissa uhattuna. Kaiken kaikkiaan tutkimukseni viittaa siihen, että lapinvuokon pölytys voi lyhyellä aikavälillä parantua, mutta pitkällä aikavälillä muutokset pölyttäjäyhteisössä uhkaavat lapinvuokon pölytyksen onnistumista. Lapinvuokko on arktisen alueen tärkeimpiä kasveja niin pölyttäjien kuin monien muidenkin hyönteisten ravintona. Siten muutokset sen menestyksessä heijastunevat koko pölyttäjien ja kukkakasvien muodostamaan yhteisöön.
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(2018)Mitochondrial dysfunction is involved in many neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS) and Huntington's disease (HD). Most of the mitochondrial disorders lack effective treatments so far. One of the important defects of mitochondrial dysfunction is the blockade of mitochondrial respiratory chain. An alternative oxidase (AOX), originated from Ciona intestinalis, has been found to counteract this defect by providing a bypath of the mitochondrial respiratory chain in plants, drosophila, mice and human cultured cells. Therefore, we suggest that AOX might be a promising tool for curing neurodegenerative diseases from the aspect of alleviating mitochondrial defects. An engineered mouse model has been created to express the Ciona intestinalis AOX ubiquitously in the whole animal. Our study aims at investigating the expression and function of AOX in the transgenic mouse brain and exploring therapeutic potential of AOX in human diseases. AFG3l2 gene encodes a subunit of m-AAA metalloprotease, which is involved in mitochondrial protein quality control process. Mutations in the AFG3L2 is known to be related to spinocerebellar ataxias 28 (SCA28), a neurodegenerative disease resulting from cerebellar damage. Loss of AFG3L2 gene causes OPA1 protein cleavage and mitochondrial fragmentation which are easily detected. Thus, we propose an in vitro neurodegenerative disease model generated by knocking down of AFG3L2 in target cells. We used western blotting to detect the expression level of AOX in the brain and primary neuro-glia cells of the transgenic mice, finding that the AOX protein reduces in the brain along with age. Antimycin A treatment on neuro-glia cells demonstrated the function of AOX in transgenic pups. However, the respiratory activity caused by AOX also decreases with the mice age. In addition, we constructed a lentiviral vector with DNA recombinant technology to express AOX in U2OS cells. To examine the function of AOX under disease condition, we established an in vitro neurodegenerative disease model by knocking down the AFG3L2 gene of U2OS cells with siRNA, and infected the cells with AOX lentivirus. By comparing the immunostaining images of wild-type and AOX infected cells, we saw a significant difference in the degree of mitochondrial fragmentation between these cells. Nevertheless, the cleavage pattern of OPA1 protein, which is mainly responsible for mitochondrial fragmentation, remains unchanged whether AOX is present in the cell or not. In conclusion, our study first investigated the expression and function of AOX in the brain of this transgenic mouse model. And we established a lentivirus vector to apply AOX gene to human cells, testing the therapeutic function of AOX in a neurodegenerative disease model in vitro. Moreover, the mitochondrial fragmentation of the disease model is alleviated by AOX despite of OPA1 cleavage, indicating that the mitochondrial phenotype caused by loss of AFG3L2 is not only due to OPA1 cleavage, but also related to respiratory defects. This highlights the possibility to rescue the mitochondrial morphological changes from new aspects.
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(2012)In multicellular organisms, complex signalling mechanisms have evolved to guide the behaviour of individual cells. Growth factors are secreted proteins that can stimulate the proliferation and/or differentiation of cells. Vascular endothelial growth factor D (VEGF-D) is a ligand for VEGF receptor 2 (VEGFR-2) and for VEGFR-3, which are predominantly expressed on blood vascular endothelial cells and on lymphatic endothelial cells, respectively. Thus VEGF-D can contribute to growth of both blood vessels (angiogenesis) and lymphatic vessels (lymphangiogenesis). Although there have been many reports showing the angiogenic and lymphangiogenic effects of VEGF-D, its physiological role is still largely unknown. Most of these reports are severely hampered by incomplete characterization of the specific form of VEGF-D that was used. During or after secretion, VEGF-D undergoes complicated proteolytic processing. Alternative Nterminal cleavage results in two different fully processed forms, VEGF-D major and VEGF-D minor. Processing significantly increases the activity of VEGF-D towards its receptors. Surprisingly, it is still unknown whether the differential N-terminal cleavage of VEGF-D has any effect on receptor binding activity or on receptor activation. The goal of this study was to produce and purify high quality biologically active VEGF-D which is needed for studying the physiological role of this growth factor. Several different forms of recombinant human VEGF-D were produced using the Drosophila Schneider 2 insect cell system. A bioassay utilizing the Ba/F3 cells expressing chimeric VEGFR/EpoR receptors was used to determine the receptor binding activities of recombinant VEGF-Ds. Two constructs producing biologically active VEGF-Ds were chosen for chromatographic purification (untagged major and his-tagged major forms). During purification, the activity of both VEGF-D forms towards their receptors decreased significantly. In case of the untagged form, this was presumably due to some residual proteolytic activity during purifications. The results might indicate that only the major form is responsible for the activation of VEGFR-3. The fact that no activity of the minor forms was detected when screening the cell supernatants with Ba/F3-VEGFR-3-EpoR-bioassay, supports this explanation. If this explanation can be verified, the role of the alternative N-terminal cleavage becomes obvious: By proteolysis the activity of VEGF-D can be redirected from the lymphatics towards the blood vessels.
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(2015)Autophagy is a eukaryotic cellular process where intracellular material is recycled by transporting it in newly formed vesicles to lysosomes for degradation. In normal conditions autophagy supports cellular homeostasis. Different stress conditions can induce autophagy and then it helps the cell to avoid an unnecessary or uncontrolled cell death. RAB proteins are small GTPases that regulate vesicle traffic and fusion events in endocytic and exocytic pathways. RAB24 has recently been shown to participate in autophagy, but there is very little information about how it works at the molecular level. GOSR1 is a Golgi SNARE protein that regulates membrane fusion events, and it has been observed to interact indirectly with RAB24. The participation of GOSR1 in autophagy has not been studied yet. The aim of the study was to find out if RAB24 and GOSR1 colocalize into the same vesicle structures and if they interact within each other. HeLa cells were used as a model organism, and to induce autophagy amino acid starvation was used. For GOSR1 detection a DNA construct was created where GOSR1 was tagged with green fluorescent protein GFP-sequence. Localization was studied with immunofluorescence staining where in addition to RAB24 and GOSR1 also the autophagosomal marker protein LC3 was labeled. The labeled cells were photographed with a confocal microscope. The pictures were analyzed with ImagePro software. Interaction between the proteins was studied using immunoprecipitation. GOSR1 and RAB24 were not observed to colocalize into same structures in significant amount. Instead it was found that GOSR1 colocalized into LC3-positive autophagic vesicles. In immunoprecipitation studies no interaction between RAB24 and GOSR1 could be shown. In order to ensure the results more immunofluorescence stainings should be done using several time points and GFP-tagged GOSR1. Also GOSR1 silencing with siRNA should be used in order to find out if GOSR1 is necessary for autophagy. The immunoprecipitation protocol should be optimized, and the possible interaction could be studied by using other methods, for example yeast-two hybride technology.
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(2013)Työssä tutkittiin rasvahappojen kerrostumista itämerennorpan (Phoca hispida botnica) traanissa ja tämän merkitystä rasvahappokoostumukseen perustuvassa ravintokohteiden arvioinnissa. Vertaamalla sisätraanin rasvahappokoostumusta plasman ja maksan rasvahappokoostumuksiin pyrittiin selvittämään siirtyvätkö jotkut tietyt ravinnon rasvahapot toisia tehokkaammin traanin sisäosiin, eli heijastelevatko jotkut sisätraanin rasvahapoista ravinnon rasvahappokoostumusta toisia paremmin. Itämerennorpan traanin eri kerrosten rasvahappokoostumusta verrattiin myös sen tärkeimmän ravintokalan, Itämeren silakan rasvahappokoostumukseen, jotta nähtäisiin minkä kerroksen koostumus muistuttaa eniten ravinnon rasvahappokoostumusta. Vertailun vuoksi työssä tutkittiin myös makeassa vedessä elävän saimaannorpan (Phoca hispida saimensis) traanin ja maksan rasvahappokoostumuksia. Lisäksi määritettiin plasma- ja maksanäytteiden kuljettaman tai lyhytaikaisesti varastoiman varastorasvan määrät. Tutkimuksessa käytettiin Perämerellä ammutuista itämerennorpista kerättyjä traani-, maksa- ja plasmanäytteitä, sekä kuolleina löydetyistä saimaannorpista kerättyjä traani- ja maksanäytteitä. Kudosnäytteiden rasvahappokoostumusten määritys tehtiin analysoimalla niistä valmistettuja rasvahappojen metyyliesteriseoksia kaasukromatografisesti (GC). Lipidiluokkakoostumukset puolestaan määritettiin korkean erotuskyvyn ohutlevykromatografialla (HPTLC). Analyysien tuloksia käsiteltiin tilastollisesti pääkomponenttianalyysin (PCA) ja sen tuloksia ohjatusti luokittelevan menetelmän (SIMCA) avulla, regressioanalyysillä, sekä laskemalla koostumusten euklidisia etäisyyksiä eri näytteiden välillä. Jokaisen yksilön traanille luotiin vertikaalinen rasvahappoprofiili toisiaan nahasta lihakseen seuraavien osanäytteiden rasvahappokoostumuksen perusteella. Itämerennorpan traanin kerrostuneisuutta tutkittiin nyt ensimmäistä kertaa ja sen havaittiin olevan rakenteeltaan kerrostunut, kuten on havaittu myös aiemmin tutkituilla kahdella norpan alalajilla. Aiemmista tutkimustuloksista poiketen keskitraani ei kuitenkaan eronnut rasvahappokoostumukseltaan merkitsevästi muista traanikerroksista. Sisä- ja ulkotraanin väliset rasvahappokoostumuksen erot olivat sen sijaan merkitseviä. Traanikerroksista sisätraani muistutti eniten itämerennorpan tärkeän saalislajin, silakan, rasvahappokoostumusta. Itämerennorpan kudosten rasvahappokoostumus erosi selvästi saimaannorpan kudosten rasvahappokoostumuksista. Näiden kahden alalajin ulkotraanit kuitenkin muistuttivat toisiaan rasvahappokoostumukseltaan sisätraaneja enemmän, mikä viittaa siihen, että niiden ulkotraanin koostumusta säätelevät samankaltaiset lämmönsäätelyyn liittyvät geneettisesti määräytyvät tekijät. Rasvahappokoostumusten alalajikohtaisista eroista huolimatta traanin rasvahappojen kerrostumistapa oli samanlainen molemmilla tutkituista alalajeista. Traanin vertikaaliset rasvahappoprofiilit olivat kuitenkin hyvin yksilöllisiä. Useiden rasvahappojen suhteelliset määrät plasmassa ja sisätraanissa korreloivat tilastollisesti merkitsevästi keskenään. Tämä tulos vahvistaa oletuksen, että viimeaikaisella ravinnolla on vaikutusta erityisesti sisätraanin rasvahappokoostumukseen ja, että sen perusteella voidaan saada tietoa eläimen ravinnosta. On kuitenkin huomattava, että tietyt sisätraanin rasvahapot ilmentävät ravinnon rasvahappokoostumusta toisia paremmin. Vaikka traanien vertikaaliset rasvahappoprofiilit antavat mitä ilmeisimmin yksilökohtaista tietoa eläinten ravinnosta ja aineenvaihdunnasta, niitä ei ole pystytty tulkitsemaan aiemmissa tutkimuksissa kovinkaan syvällisesti. Nämä tulokset voivatkin osaltaan auttaa tulkitsemaan traanin vertikaalisia rasvahappoprofiileja tulevaisuudessa. Saatuja tuloksia voidaan lisäksi hyödyntää myös hylkeiden ravintokohteiden arvioinnissa käytettävien näytteenottoprotokollien suunnitteluun ja kehittämiseen.
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(2015)Microbiota, the microbial communities living in the gut, skin and glands of vertebrates, is the functional link between a physiological individual and it s surrounding ecosystem. Mutualistic microbes play a role in social systems as well, since they are transmitted through social behaviors and claimed to affect host sociality and behavior. Furthermore, individual stress physiology can affect both behavior and microbiota. I took first steps to reveal the role of gut microbiota in the social dynamics of a group-living cooperative primate, the red-bellied lemur (Eulemur rubriventer). Gut microbiota was affected by seasonal change, stress hormonal profile and pregnancy, and individuals were found to have a strongly group-specific gut microbial composition, highlighting the role of social environment in determining gut microbial composition. Unlike expected, individual sociality was negatively associated with gut microbial diversity. Gut microbiota seems to be largely determined by social factors in this species, indicating that social transmission of beneficial microbiota might have played a role in the evolution of sociality.
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(2017)Chitinolytic enzymes belong to a group of pathogenesis-related proteins, which are induced in plants by a pathogenic attack. They have also been shown to function in abiotic stresses and related to signalling. Chitinases catalyze the degradation of β-1,4-N-acetyl glucosamine units of chitin. Chitin is not found in plants, but for example the shells of invertebrates, cell walls of certain fungi, algae and bacteria contain it abundantly. Lately chitinases have gathered attention because of their potential utilization possibilities in medicine, agriculture, food industry and biofuels. Strong chitinase gene expression and also chitinolytic activity has been found in a spruce (Picea abies L. Karst.) cell culture, which produces apoplastic lignin into the medium. Plant chitinases are known to be induced also in developmental processes. Chitinase-like genes have been shown to be essential in cellulose synthesis and in development of cell walls and tracheary elements and also affected the localisation of lignin in tissues. Therefore, the secretion of chitinases into the culture medium has not necessarily been caused by stress and chitinases might use also some endogenic substrate in addition to chitin. Because chitin is a structural component of plant pathogens but not present in plants and the growing circumstances of the studied aceptic spruce cell culture were stable, the research hypothesis was, that the secreted chitinases in the culture medium were not a stress reaction but related to development and besides chitin of the pathogens they utilize some own polysaccharide substrate of spruce. Additional hypothesis was connection of the strong expression of chitinases and chitinase-like genes in the culture medium and the production of apoplastic lignin by the spruce cell line. The aim of the study was to characterize the chitinases and chitinase-like enzymes of the spruce cell suspension culture medium and developing xylem by isolating, purifying and cloning them and by producing them heterologously. The aim was also to identify the putative endogenous substrates for the chitinases and chitinase-like proteins to reach better understanding about the meaning of the chitinases in the development of the spruce tissues. Two chitinases of the spruce cell suspension culture were isolated and purified by chromatographic methods. One chitinase gene in spruce cell culture and one chitinase-like gene in spruce xylem were cloned, after which the chitinases and chitinase-like proteins were produced in Pichia pastoris yeast. The total proteins of the culture medium, the purified chitinases isolated from the medium and the chitinases and chitinase-like proteins produced in P. pastoris were used to examine their substrate alternatives with the carbohydrate microarray method. The connection between apoplastic lignin and the chitinolytic enzymes in culture medium was not found and obvious candidates for the endogenous substrates were not detected. However, the mannan degradation in the presence of the chitinases was strong enough to give reason to further analyses.
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(2014)Havsekosystemen runt om i världen lider av antropogen inverkan såsom näringsutsläpp och utfiske. Speciellt utfiske av högre trofiska nivåer dvs. stora rovfiskar, anses ha ändrat dynamiken i flera näringsvävar. Ändringarna har lett till kraftiga trofiska kaskader som ökat eller minskat primärproduktionen speciellt vid kustliga vattenområden. Speciellt mesopredatorer har ökat i antal, och i Östersjön är storspiggen (Gasterosteus aculeatus) en art som börjat dominera samhället. Storspiggen konsumerar växtavbetare av trådalger, och kan därmed ha en effekt på trådalgstillväxten i Östersjön. Undersökningens syfte är att granska vilken roll storspiggens predation har i trådalgsbältet, och hur ändringarna syns i näringsväven. Undersökningen bestod av ett mesokosmexperiment och en fältundersökning vid Tvärminne zoologiska station. Mesokosmexperimentet bestod av två behandlingar: 10 mesokosm med två trofinivåer (utan storspigg), och 10 mesokosmer med tre trofinivåer (med storspigg). Trofinivåerna bestod av trådalger (Cladophora sp.), märlkräftor (Gammarus sp.) och storspigg. Experimentet pågick 4 veckor, vartefter mesokosmerna tömdes och innehållet analyserades. Med en veckas mellanrum gjordes fältbesök till den närbelägna ön Långskär, för trådalgsprovtagning, observation av storspiggstäthet och magsäcksanalysering av fångade storspiggar. Trådalgsproverna analyserades under mikroskop. Fältundersökningens resultat visade att mängden storspiggshanar och fångade storspiggar minskade längs med säsongen, medan trådalgsbiomassan ökade. Antalet vuxna storspiggar korrelerade negativt med märlkräftor. Mesokosmexperimentets resultat visade att mesokosmen med storspigg hade i medeltal högre trådalgs-biomassa (24%), högre epifytmängder (72%), färre märlkräftor (79%) och lägre färgintensitet än mesokosmerna utan storspigg. I Mesokosmerna utan storspigg hade trådalgsbiomassan inte ökat nämnvärt, men trådalgen var tydligt grönare i färg och saknade epifyter. Av resultaten framgår att storspiggen påverkar primärproduktionen i ekosystemet, genom en trofisk kaskad, då växtavbetarna regleras kraftigt. Primärproduktionen ökar mest i form av epifyter och mikroalger, vilket leder till att trådalgen kvävs och börjar föråldras. Trådalgen och växtavbetarna har därmed ett mutualistiskt förhållande. För att minska mängden skadliga algblomningar i Östersjön, borde näringsvävarnas stabilitet och mångfald tas i beaktande vid fiskeriförvaltningen för att effektivera Östersjöns rehabilitering.
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(1998)Cells in tissues have only three serious options in life; they can grow and divide, remain static, or die by apoptosis. Upon growth factor stimulation a cell enters the so called cell cycle which will eventually lead to the division of the cell. Cell cycle can be divided into four phases; G1, S, G2 and M. The current model of the cell cycle control holds that the transitions between different cell cycle states are regulated by cyclin dependent kinases (CDK) with their activator subunits, the cyclins. CDK regulation can be separated into four distinct mechanisms, one of which being phosphorylation on the so called T-loop leading to complete activation. This phosphorylating activity is mediated by apparently a single enzymatic activity termed the CDK activating kinase, CAK. CAK activity was originally isolated as a biochemical purification extract and the enzyme was surprisingly noticed to be structurally related to CDKs. Since a novel cyclin was identified to be associated to it, the enzyme exercising CAK activity was named CDK7 and the cyclin was designated cyclin H. An entirely new perspective on CDK7 function was opened when CDK7 was identified as a subunit of transcription factor IIH (TFIIH) and shown to phosphorylate the carboxy-terminal domain (CTD) of RNA polymerase II (RNAPII). CDK7 has also been suggested to be involved in irradiation sensitivity pathways and nucleotide excision repair functions. To elucidate the intriguing in vivo role of CDK7, proteins interacting with CDK7 were screened for using the yeast two-hybrid method as part of previous studies of the laboratory. The results showed that 15 out of 144 (10,4%) positive clones were identified to encode a peptide sequence of a protein previously known as the inhibitor/interactor of protein kinase C (PKCI). These yeast colonies had an unexpected phenotype; contradictory to a dark blue color of the colonies, indicating strong interaction, the size of the PKCI colonies was small compared to others, indicating a possible growth inhibition effect. Several DNA open reading frames (ORF) coding for proteins related to human PKCI have been identified in a broad range of species representing mammalian, plant, fungal and bacterial kingdoms, all these forming a HIT (conserved triad of histidines) protein family. Another human member, part of this now super family, named FHIT (fragile triad of histidines) was identified with a dinucleoside 5’,5’’’-P1,P3-triphosphate hydrolase activity. These molecules; substrates of FHIT and related enzymes have been proposed to have various intracellular functions, including signalling stress responses. The aim of this study was to extend the investigation of the interaction between CDK7 and PKCI observed in yeast two-hybrid by means of several genetic and biochemical approaches to determine if this observed interaction and growth phenotype has any physiological significance. Investigations included performing yeast two-hybrid screening for PKCI, developing yeast three-hybrid system and carrying out growth rate assays for yeast liquid cultures. These studies also included performing biochemical purifications of over-expressed proteins, immunoprecipitations, western blot analysis and kinase activity assays. Protein extracts originated from transformed yeast cells, transfected mammalian cells or from in vitro transcription and translation reactions. On basis of growth rate assays it can be concluded that PKCI has an inhibitory growth effect in yeast. The preliminary finding of a specific PKCI-CDK7 interaction in yeast two-hybrid, however could not be conclusively verified by the other methods that were used in this study. Studies of PKCI characterisation also included examination of the subcellular localisation of PKCI in mammalian cells by immunofluorescence labelling of HA-PKCI. Results showed PKCI to localize both in the nucleus and in the cytoplasm. Also, studies to elucidate the function of PKCI were performed; whether it possesses enzymatic activity related to that of FHIT. By NMR spectroscopy using bacterially produced GST-PKCI, hydrolase activity towards ADP was indeed observed. Future studies will include elucidation of possible links between growth inhibition and hydrolase activity, in the form of stress signalling functions. The main focus of our future studies will be the generation of mice with targeted PKCI alleles offering powerful means to reveal the function of PKCI through observing phenotypes and through detailed analysis of these mice harbouring wild type, hypomorphic or null alleles.
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(2012)The purpose of this master's thesis is to study nature values within the Finnish national Natura 2000 network on state owned land. The six goals of this work were achieved: 1) Areas with most nature values were identified by prioritizing habitats of Natura 2000 directive (92/43/EU) within. Areas with high nature value were usually in very natural state and had good connectivity to other similar places, or they were spots of some very rare nature types. 2) It was found out that data used was suitable for identifying conservation values, 3) find out the suitability of Zonation software in conservation area management and maintenance planning and 4) find out how results will change if conservation status is taken into account. As an addition to these 5) the most considerable areas with high conservation value were identified and 6) "Zonation software in a nutshell" was produced in Finnish to assist Finnish state officials to use the software for conservation purposes. These results will help Metsähallitus (The Finnish Forest and Park Service) - Natural Heritage Service - to target resourcing of habitat management and restoration in and around the areas with most considerable nature values. It is essential to sustain these areas and their values so that their nationwide importance can be maintained into the future. Data used in this study covered areas that were classified as Natura 2000 habitats according to European Union Council Directive 92/43/EEC. Analyses were done by using Zonation software, a tool for spatial conservation prioritization. Data consisted of 68 Natura 2000 habitat types and their state of naturalness and representativeness. Zonation took into account the rarity, quality, importance, threat status, biodiversity value, congruity and connectivity of these habitat types. As a result software produces a map of conservation priorities and associated quantitative information, which facilitate identification of areas with most considerable nature values. These were identified both ocularly and with Zonation software. Analyses were done at two levels: all habitat types together and in subgroups following division to major habitat types, such as coastal environments, inland waters, meadows, alpine habitats, peat lands, rocky areas and forests. Results showed that connectivity increased aggregation of areas with high nature values and weighting spread them. Hierarchical analysis was used to find out how nature values changed when the conservation status of the areas were taken into account. The results of hierarchical analysis show that conservation status changed the results a lot. Difference between main analysis and hierarchical analysis was much greater than when taken into account connectivity of feature weights. Hierarchical comparison revealed that many areas with considerable high nature values areas are not presently strictly protected.
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(2010)Elinympäristön vaikutusta eliölajien esiintymiseen voidaan tutkia habitaattimallinnuksen keinoin. Havainnot lajin esiintymisestä ja puuttumisesta eri kohdissa maisemaa suhteutetaan tilastollisen mallin avulla ympäristötekijöihin, jolloin saadaan kuva lajin elinympäristövaatimuksista. Tällöin oletetaan, että laji esiintyy kaikkialla siellä, missä sen elinympäristövaatimukset täyttyvät. Metapopulaatioteorian valossa näin ei kuitenkaan aina ole: maisemassa voi olla runsaasti lajille soveltuvia mutta asuttamattomia kasvupaikkoja, koska uusien yksilöiden leviäminen ja vanhojen häviäminen eivät välttämättä ole yksittäisten kasvupaikkojen tasolla tasapainossa. Siten myös lajin leviämiskyky vaikuttaa siihen, millaiseksi sen levinneisyyskuvio maisematasolla muodostuu. Tässä työssä keskityn tammen (Quercus robur) alueellisen levinneisyyden mallintamiseen. Tammen suhteellinen harvinaisuus Suomessa sekä sen rooli luonnon monimuotoisuuden merkittävänä tukipilarina tekevät siitä mielenkiintoisen kohdelajin ekologiselle tutkimukselle. Tutkimuskohteekseni valitsin Wattkastin saaren, n. 5 km2:n alueen Länsi-Turunmaalta. Wattkastissa on tutkittu kahdeksan vuoden ajan tammella elävien hyönteisyhteisöjen rakennetta ja kannanvaihteluita, ja saaressa kasvaa yli 1800 luonnonvaraista tammea, joiden tarkat sijainnit tiedetään. Tässä ympäristössä tutkin, rajoittaako tammen alueellista levinneisyyttä ensisijaisesti sopivien elinympäristöjen tilajakauma vai pikemminkin sen leviämiskyky. Yhdistin tammen esiintymiskuviota kuvaavaan habitaattimalliin kokeellisen aineiston, jonka avulla arvioin tammen paikallisen esiintymiskuvion ja tammelle soveltuvien elinympäristöjen tilajakauman vastaavuutta. Kokeellisen aineiston muodostivat 104 Wattkastiin vuonna 2004 istutettua pikkutammea, joiden selviytymisen ja kasvupaikkaolot kartoitin syksyllä 2009. Tutkin yleistetyillä lineaarisilla malleilla puiden menestymiseen - siis selviytymiseen ja kuntoon - vaikuttavia tekijöitä. Etsin potentiaalisia selittäjiä tammen menestymiselle kasvupaikan ympäristötekijöistä sekä istutetun puun sijainnista suhteessa luontaisiin tammikasvustoihin. Lisäksi tutkin habitaattimallin avulla, selittävätkö ympäristötekijät tammen nykyisen esiintymiskuvion Wattkastissa. Havaitsin, että istutetut puut olivat selviytyneet keskimäärin hyvin ja ettei niiden menestyminen riippunut sijainnista suhteessa luontaisiin tammikasvustoihin. Habitaattimallin selitysaste oli vain 19 %, eli kasvupaikkatekijät selittivät heikosti tammen nykyisen esiintymiskuvion Wattkastissa. Tulosten perusteella tammen paikallinen esiintymiskuvio ei vastaa sille soveltuvien elinympäristöjen jakaumaa maisemassa, joten tammen levinneisyyttä Wattkastissa rajoittaa ilmeisesti sen leviämiskyky. Tulokseni viittaavat siihen, ettei tammen elinympäristön laadussa ole suuria vaihteluita Wattkastin sisällä, koska sopivia kasvupaikkoja on tarjolla tammen nykyesiintymiseen nähden runsaasti. Tämä on tammihyönteistutkimusten kannalta kiinnostava tulos, koska se tarkoittaa, että aiemmat havainnot isäntäpuun sijainnin ja hyönteisten kannanvaihteluiden välisestä yhteydestä edustavat todellisia, tilaan sidottuja populaatioprosesseja eivätkä isäntäpuun välittämiä eroja elinympäristön laadussa. Lisäksi tutkimukseni osoittaa, että yhdistämällä habitaattimallinnukseen kokeellinen lähestymistapa saadaan realistisempi kuva lajin levinneisyyttä rajoittavista tekijöistä kuin tutkimalla pelkästään ympäristötekijöiden vaikutusta lajin esiintymiseen. Jos lajin rajoittunut leviämiskyky on vaikuttanut sen esiintymiskuvion syntyyn, pelkästään ympäristötekijöihin perustuva levinneisyysmalli liioittelee levinneisyyttä. Kokeellisen tutkimuksen avulla on tällaisessa tapauksessa mahdollista paljastaa myös leviämiskyvyn rooli esiintymiskuvion taustalla.
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(2012)The non-lethal effects of predation, i.e. predation risk, can significantly affect the prey population by inducing changes in behavior to reduce the risk of predation. Vigilance, hiding, and fleeing are common responses in order to lower predation risk while changes in habitat selection, habitat use, and changes in activity patterns are more severe changes and can profoundly affect prey fitness. Prey may begin to avoid habitats frequented by predators and may begin to reduce their activity during the time of day when predators are active. Human disturbance can be comparable to predation risk as it may induce similar changes in behavior. Therefore, human activity can be compared to predation risk even though the risk posed by humans may rarely be lethal. For many large vertebrates, however, humans do pose a direct and lethal threat. This is especially true for large predators whose severe decline has been attributed to centuries of persecution and habitat loss and whose populations have only recently began to increase following more favourable management plans and conservation efforts. Similarly, brown bear (Ursus arctos) populations have only recently began to increase in many parts of Europe and North America. In Finland brown bears survived extinction only in the wilderness areas in the north and in the east but have recolonized much of the country in the past decades. These solitary, opportunistic omnivores prefer forested habitats and usually try to avoid humans who they may view as predators. The limited availability of ideal habitats and extensive human activity means that bears may have to use spatio-temporal avoidance of humans rather than large scale spatial avoidance in order to reduce the risk of encountering humans. This should be evident in bear daybed selection whereby bears should select daybed sites away from human activity and select sites that provide good cover against humans while the bear rests during the day i.e. peak period of human activity. Additionally the daybed concealment should be higher closer to human activity. I studied bear daybed selection using GPS location data from collared bears from Central Finland and North Karelia regions and identifying possible daybed sites. Habitat of the daybeds was studied both by visiting daybeds in the field to assess the tree height, tree species composition, and concealment (visibility and canopy cover) and by using GIS software to identify large scale habitat preference. Effects of human activity (house, small roads, large roads) were studied by comparing the concealment of the daybed to a nearby random site and by comparing the bears' early morning locations' distance to human activity with subsequent daybed locations. Further tests were performed to test if season, bear sex, or the study area affected bear daybed selection. The results indicate that bears strongly preferred forested habitats in daybed selection and preferred mixed forests and woodland shrub habitat over the dominant coniferous forests possibly due to their higher proportion and availability of spruce (Picea abies) and deciduous trees (e.g. Betula spp. and Populus tremula) that can offer better cover than pine (Pinus sylvestris) which is dominant in coniferous forests. Mixed forests and woodland shrub may also be preferred as they provide bears with more nutritious vegetation as a food source. Both males and females preferred sites with shorter trees for better concealment while females with cubs preferred sites with taller trees with less concealment. Thus females with cubs may have to trade high concealment for better escape chances for the cubs to avoid potentially infanticidal males. Human activity was shown to affect daybed selection as daybed sites were more concealed than nearby random sites and bears were also closer to human activity during early morning but selected their daybeds farther away. Daybeds were considerably farther away from houses and large roads than from small roads which were often scarcely used forestry roads. Concealment did not change with distance to human activity indicating that bears may not be able to avoid human activity at a large spatial scale even in areas of low human population density. Due to low sample sizes and individual bias the results of this study should be considered more indicative than of high probability. Nevertheless they provide largely new information on bear daybed selection that may be of public interest in reducing human bear conflicts and of use in bear conservation and management.
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(2014)Sexual signals, that are part of animal signalling systems, are important in attracting and competing for mates. Costly sexual signals can vary in honesty. Honest sexual signals convey truthful information about the signaller's condition and quality, reflecting reliably the direct and/or indirect benefits. Eutrophication of the Baltic Sea, which arises from an increased input of nutrients into an ecosystem by human, alters the water turbidity, especially in coastal areas, which affects visual sexual signalling in the three-spined stickleback (Gasterosteus aculeatus). Boosted algal growth increases water turbidity and decreases the amount of light that penetrates the water column, and thus reduces visibility. This change in visibility can affect the honesty of sexual signals. My aim was to study if the three-spined stickleback sexual signals honestly reflect a direct benefit of mate choice, male parenting ability, when the environment changes due to increased water turbidity and if the maintenance of honesty depends on the social setting in the presence of competing males. I studied the maintenance of signal honesty by having two experimental treatments; turbid water treatment and clear water treatment. I had two males interacting in a same experimental aquarium. I showed the males a female and recorded courtship activity twice for 15 minutes. The female was then released and the male followed as well as the time it took to follow the male to the nest were recorded. I photographed the males in days one, two, three and six of the experiment to be able to observe the change in the nuptial colouration. The males were let to spawn and the egg mass was weighted for the eggs received and the eggs hatched. I found that the courtship activity tended to be reduced in turbid water. Increased water turbidity relaxed the association between female mate choice and courtship activity. Increased water turbidity had no significant effect on male red nuptial colouration or on female preference for redness but it reduced the difference between the two males in redness during courtship. Hatching success was found to be higher in clear water. The difference between the chosen and the not chosen male in hatching success tended to be reduced in turbid water. According to my results the honesty of courtship seemed to be maintained by adjusting courtship activity to reflect the lowered hatching success. On the other hand nuptial colouration was not adjusted and was not honest. The hierarchy between males was also weakened in turbid water which gave males an advantage to cheat. I showed that, as in many other species, such as in different bird and whale species, environmental change influences three-spined stickleback signalling and reproduction outcome. It remains to be seen whether sticklebacks will adopt an alternative signalling method, such as size or olfaction, or adjust the present visual signalling system to turbid conditions. Future will show if the changed signalling system can maintain sexual selection of good parental ability. More research is needed to understand the effect of turbidity on reproduction and population viability and uncover the broader influence of changes in the stickleback ecology to the Baltic Sea habitats.
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(2018)Host factors play crucial roles in virus infections. Viruses exploit various cellular processes and are counteracted by an arsenal of host antiviral defenses. Characterization of these interactions is crucial for understanding the viral life cycle and developing novel antiviral treatments. Semliki Forest virus (SFV) is a positive-strand RNA alphavirus that has been used as a model virus for multiple clinically significant diseases such as lethal encephalitis. The aim of this thesis was to identify host factors that affect SFV infection to better understand the biology of SFV, and to provide candidate targets for therapies against more serious alphavirus infections. Here I have conducted follow up studies on a previously performed genome-wide siRNA screen that hinted that a number of genes have novel functions in SFV infection. I used an automated high-throughput imaging-based approach to confirm the roles of these host factors in SFV infection. For comparison, I also used a similar strategy to test if these genes affect negative-strand RNA virus infections, using vesicular stomatitis virus (VSV). Additionally, I studied whether the host factors affecting SFV infections perform their roles in the entry and penetration, or post-penetration steps using a previously developed endocytic bypass assay. I identified the γ-aminobutyric acid (GABA) transporter, SLC6A13, as a potential receptor for SFV. I also describe other novel genes that have roles in SFV or VSV infections. In addition, I show that TNP01, RPL18, ETF1, DMN2, and GNDPA1 promote, and HDAC6 counteracts SFV infection in the entry and membrane penetration steps. Furthermore, I report that in the later stages of the infection DDX54 boosts and EIF2B3, EIF4G1, PHB2, EDF1, DDX47, and DHX57 hinder SFV.
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(2015)Neurotrophic factors are essential for the development of the central nervous system. By signalling through Trk receptors, they have multiple effects on for example the survival of neurons, growth of axons and dendrites and stability of synapses. BDNF, which signals through TrkB receptor, is a neurotrophic factor with an important role in the formation and stabilization of glutamatergic synapses. It is also known to be released in an activity dependent manner. The mechanisms by which BDNF and TrkB signalling regulate synaptic transmission depend on the type and developmental stage of the synapse, and they are not well known. The electrical activity of immature networks consists of intrinsic activity with intermittent bursts of synchronous activity, which is believed to fine tune the synaptic connectivity through Hebbian plasticity mechanisms, which are stabilized by homeostatic mechanisms. Homeostatic regulation can be especially important during the development of the neural network while the glutamatergic transmisson is very labile. Little is known of the signalling routes that participate in the homeostatic plasticity during the development of the neural network. The aim of the thesis was to investigate how the long-term inhibition of TrkB signalling affects the glutamatergic transmission and the homeostatic regulation in area CA3 of neonatal hippocampus by using gene manipulated TrkBF616A mouse strain. The TrkB receptors in the TrkBF616A strain are modified so that they can be blocked with a kinase inhibitor (1NMPP1). In part of the work C57BL/6 -mice were used as control. In addition to the acute measurements the hippocampal slices were incubated in control conditions and with inhibitory drugs (TTX and 1NMPP1) for 15 to 20 hours, after which miniature excitatory postsynaptic currents (mEPSCs) were recorded with whole-cell patch clamp from area CA3 pyramidal cells. The research shows that continuous TrkB signalling is essential for the maintenance of AMPA receptor mediated synaptic transmission in CA3 area of neonatal hippocampus in TrkBF616A mice. Long-term inhibition of TrkB signalling decreases the amplitude of mEPSCs. TrkB-signalling seems to be needed also for the homeostatic response caused by network activity deprivation. The results also indicate that the inhibition of TrkB signalling increases the frequency of mEPSCs, possibly by a homeostatic mechanism. However, the results also show that the TrkBF616A mice strain might differ from wild type mice and the kinase inhibitor 1NMPP1 might have non-specific effects that are not currently known, so more extensive research on the matter is still needed to confirm the results.
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