Browsing by department "Department of Biosciences"
Now showing items 21-40 of 64
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(2016)Parasitoid host interactions are intimately associated with food web dynamics as well as with evolutionary change. The parasitoid s possibility to expand its host range or switch to different hosts is affected by parasitic strategy. Strong specialization to a narrow host repertoire can arise e.g. if the larva spends considerable time in close contact with an active, developing host (koinobiosis). In contrast, an ectoparasitic larva, growing rapidly on a sessile host, does not have to be equally tightly adapted to host physiology. This may permit a wider host range. However, there are other restricting factors, such as egg production capacity. Extant parasitoids show a staggering species diversity, most notably among the Hymenoptera. Theories concerning speciation in association with host range have been developed, but draw from knowledge of merely a small part of this diversity. This study adds information by showing a number of radical host shifts in the evolutionary history of the ichneumonid wasp genus Gelis. Species of this genus exploit either insect prepupae or spider eggs. These are similar in being rather defenceless, aside from a protective silk covering. Nonetheless, differences are large enough to restrict Gelis species to each utilise only one of the two groups. In order to trace ancient host group shifts, host data was mapped onto a phylogeny constructed through bayesian analysis of sequence data. Sequences from the COI (mitochondrial) and ITS2 (nuclear, noncoding) regions were used. The results suggested at least four host group shifts within the genus. In addition, wing reduction was found to have occurred in two different lineages. Through COI barcode sequence clustering supported by morphological traits, a previously undescribed species from the G. bicolor-species complex was detected. The phylogenetic analyses also indicated the existence of further cryptic species, and the genus Thaumatogelis Schwarz was found to be nested within Gelis.
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(2015)Studies of population structure are motivated by the need to understand population history and to have well-characterised groups of individuals in studies of genetics of diseases and traits. A standard method to analyse genetic population structure is principal component analysis (PCA). A disadvantage of PCA is that it can reliably handle only independent genetic markers. This means that the genetic markers that are correlated with other genetic markers have to be excluded from the data. This leads to a loss of information. In 2012, Lawson et al. published a chromosome painting method that can utilise haplotype information, i.e. information from correlated markers, and thus it can detect more subtle differences in populations than the standard PCA. This thesis studies two questions. The first question is whether the chromosome painting method can provide more precise genetic clustering of geographically defined Finnish groups than the standard PCA method. The second question is whether the chromosome painting method can reveal new details of population structure in Finland. The data used in this study are from the FINRISK Study survey of 1997. This cohort includes the genotype data of about 4,000 individuals and the information about individuals and their parents birthplaces. 345 Individuals were randomly chosen from the cohort in such a way that both of their parents were originated from the same province. Ten provinces of Finland were used as study groups for the method comparison. First, the data were analysed with SmartPCA (a standard PCA method) and ChromoPainter (the chromosome painting method) and the results were compared both visually and quantitatively. Finally, the individuals were assigned to populations based on the ChromoPainter result using FineSTRUCTURE program and these genetic populations were compared to the geographic origin of the individuals. The results showed that the chromosome painting method clustered seven out of ten groups significantly tighter than the standard PCA. Nevertheless, SmartPCA was faster and easier to use than ChromoPainter. The main population genetic division was found between the eastern and western parts of Finland, which was consistent with earlier studies. All in all, 15 populations were detected and the results revealed that they were geographically clustered. The genetic populations correlated well with the borders of Finnish provinces and counties. As the first conclusion, the chromosome painting method was able to give more precise results than the standard PCA but the standard PCA is still more suitable for quick preliminary analyses of genetic data. As the second conclusion, the chromosome painting method was able to detect detailed subpopulation structure in Finland and these populations are geographically clustered. Results provide an excellent basis for the future studies of population structure and genetic diseases in Finland.
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(2013)The next-generation sequencing (NGS) platforms create a large amount of sequence in short amount of time, when compared to first generation sequencers. An overview of the NGS platforms is provided with more in-depth look into Illumina Genome Analyzer II as that is used to create the data for the thesis. There were two main aims in this thesis. First, to create a pipeline which can be used to analyse genomic sequencing. Second, to use the pipeline to compare whole human exome capture methods from two manufacturers, Roche Nimblegen and Agilent. The pipeline is describe in detail in material and methods. All the inputs for the pipeline are described and examples shown. In the pipeline the given sequences are first aligned against the reference genome. Then various separate analysis is performed to retrieve variants and coverage of the sequencing. Supplementary results include paired-end anomalies, larger insertion and deletion polymorphisms and assembly of non-aligned sequences. The two capture methods are also described and changes to the manufacturers' recommended protocols are listed. Finally, the section has the options and various inputs used in the pipeline runs of the exome data. The results of the pipeline is a basic level of analysis of the sequencing as well as various graphs showing the quality of the run. All the output files intended for user are described. By using the results of the pipeline, the user can do more in-depth analysis as required by the project. When comparing the two exome capture methods, the Nimblegen capture was shown to be more efficient in capturing the CCDS exome. While the Agilent capture kit provided better one fold coverage over the exome, higher fold coverage (over 10 fold), which is required for reliable variant calling in nextgeneration sequencing, was better reached using the Nimblegen capture kit. Also, significantly fewer false positive paired-end anomalies were observed in the library created by using the Nimblegen capture.
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(2017)Lactobacilli are gram-positive lactic acid bacteria with wide beneficial properties for human health and food production. Today most of the fermented products and probiotic foods are produced by lactobacilli species. One of the most using area of lactobacilli species is fermented products especially dairy products. Lactobacilli species can be used as starter or adjunct cultures in dairy products and play important role for preservation and quality, texture and flavor formation. Additionally, probiotic properties of lactobacilli species provide several health effect for human by stimulation of immune system and protection against pathogens. Lactobacillus rhamnosus LC705 is a facultatively heterofermentative type lactobacilli which is used in production of dairy products as adjunct starter and protective culture. The complete and annotated genome sequence of L. rhamnosus strain LC705 published on 2009. Known characteristics of L. rhamnosus strain LC705 are food preservation, toxin removal and health benefits when combined with other probiotic strains. However, molecular mechanism behind these characteristics are not known or not clearly understood. To get further insight on these properties and roles in cheese ripening of strain LC705, we re-annotated genome of the LC705 with updated methods and databases, analyzed metabolic pathways of LC705, and performed RNA-seq experiment to determine gene expression changes of LC705 during warm room (25 °C) and cold room (5 °C) cheese ripening process. Several un-characterized proteins of LC705 were annotated (77) and 1197 enzyme commission (EC) numbers are added to annotation file with re-annotation of genes of LC705. More importantly, re-annotation provided us 72 new pathways of LC705 which is 35% of the entire collection of 201 pathways. Analyzes of pathways showed that genome of LC705 has responsible genes for production of flavor compounds such as acetoin and diacetyl which are provide buttery flavor to dairy products, and hydrogen sulfide which is a volatile sulfur compound that cause unlikeable odor. Additionally to flavor compounds, we defined genes that produce anti-fungus compounds and bacteriocin which provide food preservation characteristic to LC705. Determination of gene expression respond of LC705 during warm room and cold room cheese ripening process with RNA-Seq showed that central metabolism genes that responsible for lyase activity, degradation activity, disaccharides and monosaccharides metabolism are warm induced genes. The genes play role in citrate metabolism pathways were significantly down-regulated during cold room, citrate degradation pathways are critical for buttery flavor products, therefore buttery flavor compounds are produced by LC705 during warm room. Finally, during cold room ripening, the genes of LC705 that produces ethanol and acetyl-CoA from pyruvate was up-regulated, so we may say that LC705 uses pyruvate to produce ethanol and acetyl-CoA instead of lactic acid.
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(2015)Archaea are known to thrive in different kinds of extreme habitats. Halophilic archaea are found in environments where the salt concentration is high, like in salt lakes and solar salterns. The habitats of halophilic archaea have salt concentration varying from higher than sea water to the saturation of salt. In high salinity habitats, the biodiversity is typically low and archaea are dominant microorganisms. The cell density of haloarchaea can be up to 107 cell/ml.When there are no predators for archaea and other halophilic microbes, viruses are thought to be the driving agents for their evolution. To this date, 130 archaeal viruses are described and 90 of them are known infect halophilic archaea. Most of the isolated haloarchaeal viruses are head-tailed. These head-tailed viruses can be divided into three different groups by the properties of the tail structure. Those three types of head-tailed viruses are myo-, sipho- and podoviruses. The infection cycles, receptors or lysis mechanisms used by archaeal viruses are still poorly known. Few studied archaeal viruses use seemingly similar strategies to attach to the surface of the host cell, to penetrate the cell surface and to release new virions, as bacteriophages and viruses of eukaryotic cells do. Since archaeas differ so much from eukaryotic or bacterial cells, their viruses must have developed different strategies to carry out their infection cycle. Several filamentous viruses of archaea are known to attach to the pilus structures of their host cells and some of the enveloped archaeal viruses attach straight to the cell membrane of the host. These strategies are seemingly similar to those used by bacteriophages and viruses of eukaryotic cells. Strikingly different mechanism to release virions has been seen on SIRV-2, virus of a hyperthermophilic archaeon. The SIRV-2 infection induces pyramid shaped extrusions on the surface of the host cells. In the end of the infection cycle these pyramids open and release new virions from the cell. This is the first lysis mechanism described for an archaeal virus. In this work, the infection cycle of a head-tailed virus infecting extremely halophilic Haloarcula vallismortis was studied. The Haloarcula vallismortis tailed virus 1 (HVTV-1) is head-tailed siphovirus with a double stranded DNA genome. In previous studies the infection cycle of HVTV-1 is described to be lytic. The genome sequence, 3D-structure of the capsid and the major capsid protein of HVTV-1 is known. HVTV-1 infection has been seen to induce large, roundish structures on the surface of the infected cell. These structures may have a role in the course of the infection cycle. Several animal viruses are known to cause massive rearrangements in the host cell so that replication proteins, viral genome and the replication agents needed from the host are concentrated in specific locations. These rearrangements facilitates and enhances the viral replication. The roundish structures induced by HVTV-1 might play the same role as the virus factories of animal viruses do or these structures may be involved in the release of new virions in somewhat similar manner as those virus induced pyramid structures. In this study two main goals were set: 1.) To explore the adsorption of HVTV-1 in more details and see if the host cell receptor could be solved. 2.) To solve the possible role of the virus induced structure seen in the host cell and to find out, in which point of the infection cycle those structures appear. Based on this study, it can be said that the adsorption of HVTV-1 is efficient and very fast. The virus attaches to the archaella structures of the host cell and the attachment is mediated by the tail of the virus. One archaella may serve as a receptor for several viruses and they are not saturated. The intracellular phase of the infection cycle of HVTV-1 is long and the virus production starts 8 hours after infection. Virions are released when the host cells is lysed 12 hours after the infection. Two hours before the lysis, two virus induced structural changes of the cell are seen. The roundish structures at the surface of the cell and wide areas in the cytoplasm filled by a structure looking similar as lipid membranes. At the same time when the virus production begins, at least two virus induced or coded proteins are produced in the cell. The other one of these proteins was identified and it is predicted to be the virus encoded ribonucleotide reductase. Ribonucleotide reductase is an enzyme known to catalyse the synthesis of deoxyribonucleic acids from ribonucleic acids. If the ribonucleotide reductase is a part of seen structures, the timing of their appearance and the known function of the enzyme, could suggest those structures to play a role in the maturation or release of the virions. Many questions still remains and there would be lot more details to study in the HVTV-1 infection. Especially interesting would be the identification of the other virus induced protein, purifying those roundish structures and analysing them in more details.
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(2013)Tässä pro gradu -tutkielmassa on tarkasteltu aivosähkö- ja aivomagneettikäyrien amplitudien vaihteluiden vastaavuussuhteita koehenkilön suoriutumiseen audiovisuaalisten ärsykkeiden tarkkaavaisuustehtävissä. Aikaisemmista tutkimuksista tiedetään, että koehenkilön osumatarkkuus ei pysy vakiona koko tehtävän ajan, vaan on monesti jaksottunut valppauden ja herpaantumisen jaksoihin. Lisäksi osumatarkkuus koko kokeen ajalta on alhaisempi kuin lyhyen kalibraatiojakson ajalta mitattuna. Tämän intuitiiviseltä tuntuvan keskittymiskyvyn järkkymisen taustalla on esitetty olevan henkilön introspektiiviset ja mielenvaelteluun liittyvät kognitiiviset toiminnot. Ennen tätä tutkimusta on jäänyt kuitenkin osoittamatta osumatarkkuuden ailahtelun yhteys aivokuoren hermostollisen aktiivisuuden pitkällä ajalla autokorreloiviin muutoksiin lähdemallintamisella. Tämän pro gradun tutkimustulokset osoittavat, että näiden kahden lajin välillä on olemassa merkittävä korrelaatioyhteys. Lisäksi lepovaiheen aivotoiminnasta modaliteettispesifeillä tarkkaavaisuus- ja oletustilan verkoston alueilla voidaan ennustaa psykofyysisen suoriutumisen vaihteluja jatkuvan audiovisuaalisen ärsykekynnyksen tarkkaavaisuustehtävän aikana. Keskittymiskyvyn vaihtelun muutoksia hermostollisella tasolla ja näitä mahdollisesti ilmentäviä käyttäytymisen ailahteluja psykofyysisinä parametreinä, kuten osumatarkkuutena ja reaktionopeutena, voidaan luonnehtia skaalauslakianalyysilla. Ilmiön skaalaton käyttäytyminen heijastelee monimutkaisen järjestelmän taipumusta luoda sisäisiä vastaavuussuhteita eli autokorrelaatioita, jotka heikkenevät hitaammin ja ulottuvat kauemmaksi ajassa ja/tai paikassa kuin mitä alla piilevistä mekanismeista voidaan suoraan ennustaa. On havaittu, että osumatarkkuuden jaksottuminen ja spontaani aivotoiminta noudattavat potenssilain skaalauskäyttäytymistä ajan suhteen. Psykofyysisen ja hermostollisen skaalauslain mukaisen käyttäytymisen kvantifioimiseksi tässä opinnäytetyössä on käytetty vaihtelun ikkunallista autokorrelaatioanalyysiä, DFA:ta. DFA paljastaa ilmiön sisällä olevien peräkkäisten tapahtumien autokorrelaatioiden kestävyyden tarkasteluvälin kasvaessa. Skaalausluvut eli DFA-eksponentit on johdettu tässä kokeessa jatkuvan audiovisuaalisen ärsykekynnyksen tarkkaavaisuustehtävän ja levon aikana rekisteröidyistä aivosähkö- ja aivomagneettikäyräsignaalien verhokäyrästä sekä psykofyysisen osuma/huti -binäärisekvenssistä rakennetusta keinotekoisesta satunnaiskulun kaltaisesta käyrästä. Jatkuvat ärsykekynnystehtävät soveltuvat hyvin tarkkaavaisuuden top-down mekanismien tutkimiseen, koska heikoista, vain juuri ja juuri havaintokyvyn säteellä olevista ärsykkeistä seuraa verraten heikko bottom-up hermostovaste. Näin keskittymiskykyyn vaikuttavat top-down säätelymekanismit kuten motivaatio, päämäärät tai mielenvaeltelu eli spontaanilta vaikuttava aivotoiminta edustuu selkeämmin aivosähkö- ja -magneettikäyrissä. Aivokuoren kokonaisvaltaisen skaalautumisen lisäksi ollaan kiinnostuneita psykofyysisten ja hermostollisten vastaavuussuhteiden jakaumamallista tietyille aivoalueille. Mitattujen hermostollisten signaalien paikantaminen tarkalleen tietyille aivokuoren alueille aiheuttaa käänteisen ongelman, joka on ratkaistu tässä MNE -lähdemallintamisella. Lähdemallintamisen algoritmit tuottavat todennäköisimmän mallin aivokuoren alueista, joiden aktiivisuudella voidaan selittää mitatut MEEG signaalit. Mallintaminen on työn kriittinen vaihe, koska sillä yhdistetään neuroanatominen tieto fysiologisen ja psykofyysisen tiedon kanssa. Yksilötason data on käsitelty lopuksi ryhmätasolla tilastollisin menetelmin korrelaatiotulosten merkittävyyksien arvioimiseksi.
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(2013)Kosteikot ovat biodiversiteetiltään yksi maailman rikkaimmista habitaateista. Kosteikkojen ja niillä elävien vesilintujen tuottamien ekosysteemipalvelujen laajuuteen ja tärkeyteen on havahduttu vasta viime aikoina. Kosteikkoja ja niiden linnustoa uhkaavat maailman laajuisesti maankäyttö sekä makeiden vesistöjen pilaantuminen. Viimeisen vuosisadan aikana maailman kosteikkojen määrä on vähentynyt puoleen ja niiden tila on heikentynyt. Kosteikkolinnuston elinolot ovat heikentyneet ja linnusto on taantunut. Kosteikkoja rakentamalla ja kunnostamalla on voitu parantaa linnuston elinolosuhteita. Etenkin keinotekoisten kosteikkojen merkitys on ollut suuri luontaisten kosteikkojen tilan heikkenemisen myötä. Vain vähän tiedetään miten luonnollisten kosteikkojen hoitotoimet vaikuttavat linnustoon paikallisella tasolla. Niukasti tietoa on myös saatavilla päättäjille hoitotoimien kustannustehokkuudesta. Tässä työssä tutkittiin Etelä-Suomen kosteikkojen hoitotoimien vaikutuksia levähtävään ja pesivään linnustoon. Työn tarkoituksena oli selvittää eri hoitotoimien vaikutuksia ravinnonhankinnaltaan erilaisiin lintukiltoihin ja -ryhmiin. Yhtenä tutkimuksen päätavoitteena oli luoda käsitys hoitotoimien kustannustehokkuudesta sekä mihin resurssit kannattaa käyttää. Tutkimusalue käsitti 21 Etelä-Suomen linnustollisesti arvokasta kosteikkoa. Kosteikkoja kunnostettiin vuosina 2004–2012 kahdessa hoitojaksossa. Hoitojaksoja ennen ja niiden jälkeen kosteikkojen pesivä ja levähtävä linnusto selvitettiin. Suomen ympäristökeskus vastasi hoitotoimista ja linnustolaskennoista Uudenmaan ja Kymenlaakson ELY-keskusten kanssa. Kosteikkoja kunnostettiin pääasiassa avoimuutta lisäämällä laidunnuksen, puustonraivauksen, niiton ja äestyksen sekä ruoppauksen avulla. Muutolla levähtävä linnusto selvitettiin läpi muuttokauden noin viiden päivän välein suoritettujen laskentojen avulla ja pesivä linnusto selvitettiin viiden käyntikerran kartoitusmenetelmällä. Kohteiden linnut eriteltiin hoidettuihin osa-alueisiin sekä niiden ulkopuolelle jääviin hoitamattomiin osa-alueisiin. Kontrollialueina toimivat hoitamattomat kohteet ja osa-alueet. Lasketut linnut yhdistettiin ravinnonhankintansa perusteella kymmeneen kiltaan, jotka olivat puolisukeltajasorsat, sukeltajasorsat, kalansyöjävesilinnut, joutsenet, hanhet, kahlaajat, naurulokki, rantakanat ja kaulushaikara, avomaavarpuslinnut sekä pensaston ja ruovikon varpuslinnut. Kolmea viimeksi mainittua kiltaa tarkasteltiin vain pesimäaikana. Erikseen tarkasteltiin kansallisesti uhanalaisia sekä lintudirektiivin liitteen I lajeja. Hoitotoimien sekä niihin käytettyjen kokonaiskustannusten vaikutuksia kiltakohtaisiin lintumääriin tarkasteltiin lineaarisilla sekamalleilla. Syysmuuttoaineistossa kaikkien lintukiltojen määrät kasvoivat niitto- ja äestysalan myötä; laidunnuspinta-ala lisäsi hanhien ja kahlaajien lukumääriä. Kaikkien kiltojen lukumääriä lisäsi laidunala keväällä, lisäksi niitto- ja äestys - sekä ruoppausala lisäsivät kahlaajien lukumääriä. Kalansyöjien lukumäärät vähenivät ruoppausalan kasvun myötä, mutta yhteys oli tilastollisesti vain suuntaa antava. Pesimäaineistossa ruoppausala x kilta-interaktio osoittautui tilastollisesti merkitseväksi, mutta kiltakohtaisesti tarkasteltuna vaikutukset olivat suuntaa antavia; rantakanat ja kaulushaikara runsastuivat ja kalansyöjävesilinnut vähenivät. Sekä uhanalaisten että lintudirektiivin liitteen I lajien lukumääriä kasvatti laidunnus keväällä ja syksyllä. Hoitotoimiin käytettyjen kokonaiskustannusten suhteen runsastuivat syksyllä puolisukeltajasorsat, hanhet ja kahlaajat. Keväällä kokonaiskustannuksilla oli positiivinen yhteys puolisukeltajasorsien, kahlaajien ja naurulokin lukumääriin. Pesimäaineistossa kokonaiskustannukset kasvattivat naurulokin lukumääriä. Suuntaa-antavasti kokonaiskustannukset lisäsivät rantakanojen ja kaulushaikaran määriä sekä vähensivät kalansyöjävesilintujen määriä. Erityisesti huomioitavaa on, että hoitotoimilla havaittiin vain positiivisia merkitseviä tuloksia lintukiltoihin. Laidunnus oli tärkein yksittäinen hoitotoimi, mikä liittynee sen ympäristöä monipuolistavaan vaikutukseen. Hoitotoimista eniten hyötynyt kilta oli kahlaajat, mutta kaikki killat hyötyivät jostain hoitotoimesta. Hoitotoimien positiiviset vaikutukset liittynevät avoimuuden lisääntymiseen sekä karjan läsnäoloon, jotka voivat parantaa lintujen ruokailuolosuhteita sekä lisätä turvallisuutta. Kokonaiskustannukset paljastivat että naurulokki ja puolisukeltajasorsat hyötyivät hoitotoimista kokonaisuutena, sillä ne eivät poikenneet muista killoista yksittäisiä hoitotoimia tarkasteltaessa. Vähiten hyötyivät sukeltajasorsat sekä kalansyöjävesilinnut. Tutkimus osoittaa, että hoitotoimilla voidaan parantaa taantuneiden kosteikkolintujen elinolosuhteita. Koska hoitokohteet ovat Etelä-Suomen parhaimpia lintuvesiä, on hoitotoimien biologinen merkitys suuri. Vaikutukset eivät ole pelkästään kansalliset, vaan kunnostuksesta hyötyvät Suomenlahden muuttoreittiä pitkin pohjoiseen muuttavat kosteikkolinnut –aina Siperiaan asti. Yksi tärkeimmistä huomioista on, että hoidon loputtua kosteikkojen tila heikkenee. Hoitoa tulisi jatkaa, jottei saatuja hyötyjä menetettäisi. Tulevaisuudessa olisi tärkeää suunnitella mittavat hoitotoimet myös tutkimuksen kannalta, jolloin hoitotoimien vaikutuksia voitaisiin tutkia tarkemmin. Tärkeää olisi löytää keinot sukeltajasorsien ja kalansyöjien elinolojen parantamiseksi.
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(2011)The relationship between hosts and parasites is one of the most studied interactions between living organisms, and it is both universal and common in nature. Parasitoids are special type of parasites whose offspring develop attached to or within a single host organism that it ultimately consumes and kills. Hosts are arthropods and most parasitoids belong to the insect order Hymenoptera. For almost two decades metapopulation research on the Glanville fritillary butterfly (Melitaea cinxia) has been conducted in the Åland Islands, Finland. The studies have been concerned with the population dynamics, evolution, genetics, behavior, natural history and life history characteristics of M. cinxia, as well as with species interacting with the butterfly. The parasitoids of M. cinxia have been under long term studies and much has been learned about specific host-parasitoid interactions during the past decade. The research for this Master s thesis was done in the Åland Islands during summer 2010. I conducted a reciprocal transplant style experiment in order to compare the performance of host butterflies (M. cinxia) under attack by different parasitoid wasps (C. melitaearum). I used hosts and parasitoids from five origins around the Baltic Sea: Öland, Uppland, Åland, Saaremaa and Pikku-Tytärsaari. The host-parasitoid relationship was studied in terms of host susceptibility and parasitoid virulence, addressing specifically the possible effects of inbreeding and local adaptation of both parasitoids and their hosts. I compared various factors such as host defence ratio, parasitoid development rate, cocoon production rate etc. I also conducted a small scale C. melitaearum egg development experiment and C. melitaearum external morphology comparison between different parasitoid populations. The results show that host resistance and parasitoid virulence differ between both host and parasitoid populations. For example, Åland hosts were most susceptible to parasitoids and especially vulnerable to Pikku-Tytärsaari wasps. Pikku-Tytärsaari wasps were most successful in terms of parasitoids virulence and efficiency except in Saaremaa hosts, where the wasp did not succeed. Saaremaa hosts were resistant except towards Åland parasitoids. I did not find any simple pattern concerning host resistance and parasitoid virulence between inbred and outbred populations. Also, the effect of local adaptation was not detected, perhaps because metapopulation processes disturb local adaptation of the studied populations. Morphological comparisons showed differences between studied wasp populations and sexual dimorphism was obvious with females being bigger that males. There were also interesting differences among populations in male and female wing shapes. The results raise many further questions. Especially interesting were Pikku-Tytärsaari wasps that did well in terms of efficiency and virulence. Pikku-Tytärsaari is a small, isolated island in the Gulf of Finland and both the host and parasitoids are extremely inbred. For the host and parasitoid to persist in the island, the host has to have some mechanisms to escape the parasitoid. Further research will be done on the subject to discover the mechanisms of Pikku-Tytärsaari host s ability to escape parasitism. Also, genetic analyses will be conducted in the near future to determine the relatedness of used C. melitaearum populations.
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Identification of new protein-protein interactions with NUAK2 by proximity-dependent biotin labeling (2017)In this Master’s project, I have studied a mammalian serine-threonine kinase NUAK2 implicated in human disease but whose molecular functions and interacting proteins are as of yet poorly characterized. The goal was to identify new interacting proteins to increase understanding of the molecular functions and potentially link to human physiology and disease. Recent work from the host lab shows NUAK2 loss in cultured primary cells mimics loss of the tumor suppressor LKB1 which also acts upstream of NUAK2, together suggesting NUAK2 could be involved in tumor suppression. Currently, only two protein-protein interacting proteins with NUAK2 have been identified: NUAK2 is targeted to actin stress fibers by the myosin phosphatase Rho-interacting protein (MRIP), and it is involved in regulating cell contractility by affecting indirectly the phosphorylation cycle of the myosin light chain through inactivation of the myosin phosphatase target subunit 1 (MYPT1). In this project, I utilized a novel protein-protein interaction screening method that utilizes proximity-dependent biotin labeling to identify new interacting proteins with NUAK2 in human embryonic kidney cells (HEK 293). This method is based on fusing an E.Coli promiscuous biotin ligase, BirA*(R118G), to the investigated protein. The BirA*(R118G) ligase biotinylates all the proteins in close proximity of the fusion protein creating a history of protein-protein associations over time. Afterwards, the biotinylated proteins can be isolated by affinity purification methods and identified by mass-spectrometry. The screening identified the previously known interaction partners of NUAK2 indicating it was technically successful. In addition, I also identified in total 108 novel potential protein interaction partners for NUAK2. One of the top hits was Cytospin-A, a cross-linking protein between microtubules and actin cytoskeleton, supporting a role of NUAK2 as regulator of cytoskeleton. Supporting the validity of our finding, Cytospin-A depletion in mammalian cells causes defective actin-cytoskeleton reorganization, a very similar phenotype seen with NUAK2 depletion. In future studies, I will continue to investigate the specific role of NUAK2 and Cytospin-A aiming for detailed information on the function of NUAK2 in regulation of microtubules and actin cytoskeleton. Validation of some of the other identified interactions is expected to provide novel insights to the biology and role of NUAK2 in LKB1 tumor suppressor functions.
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(2011)Ewing sarcoma is an aggressive and poorly differentiated malignancy of bone and soft tissue. It primarily affects children, adolescents, and young adults, with a slight male predominance. It is characterized by a translocation between chromosomes 11 and 22 resulting in the EWSR1-FLI1fusion transcription factor. The aim of this study is to identify putative Ewing sarcoma target genes through an integrative analysis of three microarray data sets. Array comparative genomic hybridization is used to measure changes in DNA copy number, and analyzed to detect common chromosomal aberrations. mRNA and miRNA microarrays are used to measure expression of protein-coding and miRNA genes, and these results integrated with the copy number data. Chromosomal aberrations typically contain also bystanders in addition to the driving tumor suppressor and oncogenes, and integration with expression helps to identify the true targets. Correlation between expression of miRNAs and their predicted target mRNAs is also evaluated to assess the results of post-transcriptional miRNA regulation on mRNA levels. The highest frequencies of copy number gains were identified in chromosome 8, 1q, and X. Losses were most frequent in 9p21.3, which also showed an enrichment of copy number breakpoints relative to the rest of the genome. Copy number losses in 9p21.3 were found have a statistically significant effect on the expression of MTAP, but not on CDKN2A, which is a known tumor-suppressor in the same locus. MTAP was also down-regulated in the Ewing sarcoma cell lines compared to mesenchymal stem cells. Genes exhibiting elevated expression in association with copy number gains and up-regulation compared to the reference samples included DCAF7, ENO2, MTCP1, andSTK40. Differentially expressed miRNAs were detected by comparing Ewing sarcoma cell lines against mesenchymal stem cells. 21 up-regulated and 32 down-regulated miRNAs were identified, includingmiR-145, which has been previously linked to Ewing sarcoma. The EWSR1-FLI1 fusion gene represses miR-145, which in turn targets FLI1 forming a mutually repressive feedback loop. In addition higher expression linked to copy number gains and compared to mesenchymal stem cells, STK40 was also found to be a target of four different miRNAs that were all down-regulated in Ewing sarcoma cell lines compared to the reference samples. SLCO5A1 was identified as the only up-regulated gene within a frequently gained region in chromosome 8. This region was gained in over 90 % of the cell lines, and also with a higher frequency than the neighboring regions. In addition, SLCO5A1 was found to be a target of three miRNAs that were down-regulated compared to the mesenchymal stem cells.
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(2008)Jokirapu (Astacus astacus) on Suomessa alkuperäislaji. Rapuruttoa hyvin sietävää täplärapua (Pacifastacus leniusculus) on istutettu lähinnä Etelä-Suomeen vuodesta 1967 lähtien. Täplärapu on kookkaampi, aggressiivisempi ja lisääntymiskykyisempi kuin jokirapumme. Yleensä täplärapu dominoi jokirapua, kun rapujen elinympäristöt kohtaavat. Täpläravun dominoivaa otetta voidaan selittää tiettyjen käyttäytymispiirteiden ollessa korreloituneena. Tietyt käyttäytymispiirteet voidaan liittää yleisesti tulokaslajien dominoivaan levittäytymiseen uusille alueille. Tulokaslajin täytyy levittäytyä epäröimättä, lisääntyä hyvin pienestä alkupopulaatiosta huolimatta sekä runsastuessaan aiheuttaa suuria vaikutuksia tunkeutumaansa yhteisöön. Tulokaslajiin käyttäytymisessä voisi siis olla korreloituneena aktiivisuus, rohkeus ja eksploratiivisuus. Käyttäytymissyndroomalla viitataan korreloituneisiin yksilön yhdenmukaisiin käyttäytymispiirteiden eroihin, jotka ilmenevät tietyssä tilanteessa ja/tai läpi tiettyjen kontekstien. Kontekstilla tarkoitetaan toiminnallista käyttäytymiseen liittyvää kategoriaa (esim. levittäytyminen). Populaatio tai laji voi ilmentää käyttäytymissyndroomaa. Jokaisella yksilöllä on oma käyttäytymistyyppinsä käyttäytymissyndrooman sisällä. Saman käyttäytymistyypin omaavien yksilöiden käyttäytyminen korreloi muiden yksilöiden käyttäytymisen kanssa järjestysluvullisen arvoasteikon mukaan. Käyttäytymissyndroomat voivat vähentää käyttäytymiseen liittyvää plastisuutta tai ylläpitää yksilöiden käyttäytymisen välistä vaihtelua. Pro gradu –työni tavoitteeni on tutkia joki- ja täplärapujen käyttäytymiseroja. Olen valinnut tutkimuksen kohteeksi aktiivisen, rohkean ja eksploratiivisen käyttäytymisen, joiden korrelaatioita on jo löydetty muutamilla lajeilla. Tutkin käyttäytymiserojen toistuvuutta sekä sukupuolen vaikutusta. Oletin löytäväni positiivisen korrelaation aktiivisessa, rohkeassa ja eksploratiivisessa käyttäytymisessä ravuilla (käyttäytymissyndrooma), ja oletin näiden käyttäytymispiirteiden välisten erojen olevan täpläravuilla suuremmat. Käyttäytymispiirrekokeet suoritin Viikissä kahteen kertaan käyttäytymisen toistuvuuden testaamiseksi. Käyttäytymisen toistuvuus osoittautui melko hyväksi, vaikka jälkimmäisen kokeen (=samaa käyttäytymistä mittaava koe tehty toiseen kertaan) tulokset olivatkin usein merkitsevästi erilaisia ensimmäisen kokeen tuloksiin verrattuna. Aktiivisuus toistui hyvin käyttäytymispiirrekokeiden kesken. Jokirapujen aktiivisena olo aika on suurempi kuin täpläravuilla aktiivisuuskokeessa ja eksploratiivisuuskokeessa. Täplärapujen aktiivisuus yleensä nousi jälkimmäisessä, kun taas jokirapujen aktiivisuus laski. Sukupuoleen sidottuja käyttäytymiseroja ei juurikaan löytynyt. Rohkeaa ja eksploratiivista käyttäytymistä ilmentävät kokeet osoittautuivat epäonnistuneiksi mittaamaan näitä käyttäytymispiirteitä. En voi saamieni tulosten perusteella todeta joki- tai täpläravuilla olevan käyttäytymissyndroomaa, jossa aktiivisuus, rohkeus ja eksploratiivisuus olisivat korreloituneena.
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Kannan, kasvatusympäristön ja ehdollistamisen vaikutus lohenpoikasten pedonvälttämiskäyttäytymiseen (2011)Istutetut kalat selviävät luonnonoloissa usein huomattavasti luonnossa kasvaneita lajikumppaneitaan heikommin. Laitosten kasvatusaltaissa kasvavilla kaloilla ei ole ennen istutusta tarvetta oppia luonnossa välttämättömiä taitoja. Jos istutuskalat ovat sukupolvi sukupolven jälkeen laitoskalojen jälkeläisiä, voi myös kalojen perimä laitostua, kun perimään kertyy luonnossa selviytymisen kannalta haitallisia ja laitosoloissa hyödyllisiä ominaisuuksia. Suomessa hyvä esimerkkilaji istutuskaloista ja niiden ongelmista on lohi (Salmo salar). Yksi istutusten tuloksellisuudelle merkityksellisistä kalojen taidoista on pedonvälttämiskäyttäytyminen eli se, kuinka hyvin kalat välttävät saaliiksi jäämistä. Sekä laitosoloissa kasvamisen että perimän laitostumisen on todettu voivan muuttaa kalojen pedonvälttämiskäyttäytymiseen liittyviä käyttäytymispiirteitä. On myös saatu näyttöä perimältään laitostuneiden kalojen luonnonkantaisia kaloja suuremmasta riskistä jäädä petojen saaliiksi. Luonnonkalojen ja laitoskalojen väliset erot sopeutuvuudessa luonnonoloihin ovat poikineet runsaasti tutkimuksia, joissa on selvitetty, miten laitoskalojen pedonvälttämiskäyttäytyminen saataisiin muistuttamaan enemmän luonnonkalojen käyttäytymistä. Tärkeimpiä samankaltaistamiskeinoja ovat laitosten kasvatusaltaiden muuttaminen virikkeellisiksi eli enemmän luonnonoloja vastaaviksi ja laitoskalojen kouluttaminen ennen istutusta. Tutkin kokeissani perimän laitostumisen, virikkeellisen kasvatusympäristön ja klassisen ehdollistamisen vaikutuksia yksivuotiaiden lohenpoikasten pedonvälttämiskäyttäytymiseen. Käytössäni oli Simojoen populaation lohenpoikasia neljästä eri kanta-kasvatusympäristö-käsittelystä: perinteisissä kasvatusaltaissa kasvatettuja luonnonkannan kaloja, virikkeellisissä kasvatusaltaissa kasvatettuja luonnonkannan kaloja, perinteisissä kasvatusaltaissa kasvatettuja laitostuneen kannan kaloja ja virikkeellisissä kasvatusaltaissa kasvatettuja laitostuneen kannan kaloja. Ensimmäisessä kokeessani vertailin eri käsittelyjen kalojen poistumisnopeutta ns. lähtölaatikosta sekä uintikäyttäytymistä kaukalossa, jossa ne eivät olleet aiemmin olleet. Toisessa kokeessa tarkkailin, miten ns. hälyaine vaikutti rohkeuskokeessa olleiden kalojen käyttäytymiseen, kun niitä uitettiin uudestaan samoissa kaukaloissa. Kolmannessa kokeessa ehdollistin molempien laitoskantaisten käsittelyjen kaloja hauen hajulle hälyaineen avulla. Ehdollistamisen jälkeen tarkkailin, kuinka poikaset käyttäytyvät kaukaloissa, joihin johdettiin hauen hajua. Tein myös selviytymiskokeen, jossa vapautin saman ehdollistamiskäsittelyn läpikäyneitä kaloja altaisiin, joissa hauet saivat saalistaa poikasia ja joihin oli kasattu poikasille suojapaikaksi kiviröykkiö. Odotin luonnonkantaisten, virikkeellisesti kasvatettujen ja ehdollistettujen kalojen olevan muita varovaisempia ja hauilta paremmin selviytyviä. Virikkeellinen kasvatus lisäsi monella käyttäytymismuuttujalla mitattuna kalojen varovaisuutta tai arkuutta. Joillain muuttujilla mitattuna virikkeellisyys vaikutti varovaisuutta tai arkuutta lisäten vain luonnonkantaisiin kaloihin Siten myös kalan kannalla oli merkitystä. Yhdessä käyttäytymismuuttujassa kannalla oli lisäksi kasvatusympäristöstä riippumatonkin vaikutus. Hälyaine vaikutti kaloihin lähinnä niiden liikkumista vähentäen. Ehdollistamisen vaikutus kaukalomuuttujiin riippui kasvatusympäristöstä: ehdollistaminen lisäsi virikkeellisissä oloissa kasvatettujen ja vähensi tavallisissa altaissa kasvatettujen uimista. Ehdollistamisella tai kasvatusympäristöllä ei ollut vaikutusta kalojen selviytymiseen haukien saalistukselta. Tulokset herättivät myös mahdollisia uusia tutkimuskysymyksiä. Tulosten perusteella voin sanoa, että kasvatuksessa käytetyt yksinkertaiset ja edulliset virikkeet ja ehdollistamismenetelmät voivat vaikuttaa kalojen käyttäytymiseen muuttamalla sitä varovaisemmaksi ja sopeutuvammaksi eli mahdollisesti joiltain osin luonnonmukaisemmaksi tai luonnossa selviytymistä auttavaksi. Erot virikkeellisen kasvatuksen vaikutuksissa luonnonkantaisiin ja laitoskantaisiin kaloihin voivat kertoa siitä, kuinka tärkeää oli, että kokeissa oli mukana molempien kantojen kaloja. Tulosten soveltamismahdollisuudet ovat lupaavat, koska tämä oli yksi ensimmäisistä koejärjestelyistä, joissa virikkeellistettyjen kasvatusaltaiden kalatiheydet olivat samaa luokkaa kuin kalankasvatuslaitoksissa tavallisesti käytetyt tiheydet.
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(2015)The International Biology Olympiad is a yearly science competition; hundreds of high school students from over 60 countries take part in it. In Finland, the students are chosen by the national biology competition and the national Olympiad training camp. The research subject of this study is Finnish training for International Biology Olympiad (IBO). About ten high school students who are interested in biology take part at each training camp. Science competitions, such as IBO and training camps, are classified as non-formal or out-of-school science education. Since there is very little scientific knowledge about non-formal biology education, especially about science competitions, the theoretical framework of this study consists of non-formal science education, the relevance of science education and the development of interest. In this study, design research methodology with three research cycles was applied to develop the course. In the study, both theoretical and empirical problem analyses were used. There were two main research questions: 1) What are the needs for the development of the training camp? 2) What kind of training is relevant for the participants? The first research question was elaborated by examining a) what kind of topics of interest the participants have, b) how the participants expect the training to be relevant for them, c) what kind of relevance the previous participants experienced, and d) what kind of effects the previous training camps had on the participants interest in biology and career choice. The features of relevant biology Olympiad training were searched in the second research question. The data was collected from pre-camp and post-camp questionnaires, post-camp interviews and a questionnaire sent for the previous camp participants. The qualitative data was analyzed by content analysis and quantitative data from the questionnaires was analyzed by statistical methods. The main results were following: 1) The biggest needs for development were diversifying the contents and balancing the workload of the camp. The participants of the training camps were interested mostly about medicine and human biology -related topics and cell and molecular biology. In addition, it was found out that the previous participants considered biology education to be individually, vocationally and societally relevant for them. According to them, the training camps were especially individually relevant and had some effects on the career choice. 2) The new training camp for biology Olympiad is especially individually relevant for the participants but it has also some vocational relevance. In the science Olympiad training, special attention should be directed towards developing vocational and societal relevance. A new model for Biology Olympiad training camps was developed based on the collected research data. The individual dimension can be improved by i) diversifying the contents of the camps, ii) including inquiry-based learning modules and by iii) taking a student-centered approach to the development process. The vocational and societal dimensions of relevance can be targeted by iv) allowing the teachers of the participants to take part at camps. The vocational dimension of relevance can be enhanced v) by organizing visits to research laboratories and companies and vi) by enabling the participants to meet university students. The results of this study can be applied to not only to Science Olympiad training but also to non-formal biology education. This research provides models for developing out-of-school biology education and its relevance. This is the first design research study about Biology Olympiad training and it opens up discussion about the relevance of science competitions and Science Olympiad training.
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(2015)Darwinian selection can be measured and investigated from gene sequences. A certain gene form favored by positive selection will become more common in the population. Detecting strong positive selection is rare, but it has been found to affect genes involved in immune defense and perception of odorants. Genes under positive selection have a possible role in speciation or adaptation. This is why chemical communication, being based on the sense of smell, is an interesting topic for measuring natural selection and positive selection in particular. Social insects, such as ants, are model organisms for chemical communication. They use chemical communication not only for finding nutrition and detecting intruders, but also in coordinating the activities of several thousands of colony members. Insects perceive odorant signal molecules with their antennae. Odorant binding proteins (OBP) and chemosensory proteins (CSP) bind and transport odorant molecules through the sensillar lymph. OBPs and CSPs are also suspected to have a role in selecting odorants. This work focuses on two OBP genes and two CSP genes to study natural selection. All of these genes are conserved among all ant species. Three of these genes, OBP1, CSP1 and CSP7, are strongly expressed in the antennae suggesting that they function in chemical communication. CSP7 also has a known function in nest mate recognition in ants and OBP1 is known to bind a queen pheromone in the honeybee. The data includes gene sequences from 7 Formica ant species (270 sequences in total). The main goals of the research were to find out 1) the extent of variation between and within closely related ant species, 2) which evolutionary forces, natural selection or random drift, are behind the variation and 3) are there systematic differences between the two social forms of ants suggesting that these genes would affect the social structure of an ant colony. The variation in the sequence data was visualized by phylogenetic, principal coordinate and fixed differences analyses. Differences between populations were studied by FST values. The evolutionary forces shaping chemical communication genes within the species and populations, were studied by McDonald-Kreitman test, Tajima s D, Fu and Li test and MFDM test. The data shows that two of the species, F. cinerea and F. exsecta, significantly differ from the other five ant species, the F. rufa group species, and that the F. rufa group species don t significantly differ from each other based on these genes and this data. This could be due to their recent speciation or crossing between the species leading to hybrids in the data. The results of the evolutionary analyses are inconsistent. However, CSP7 has the strongest indication of selection based on all of the tests. Possible selection and sequence variation was detected at a predicted transcription factor binding site in F. cinerea. This indicates that selection might affect the regulation of CSP7. In the future it would be interesting to check the true transcription factor binding sites experimentally.
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(2010)During the last century increased emissions of greenhouse gases have caused global climate warming. While the temperatures are still rising, the largest increases have been observed in spring-time temperatures. This may affect the phenology of multiple species. If the increase in temperature affects different species differently, then the lifecycles of various species may become unsynchronized. For interacting species, a disturbance like this may be catastrophic: for example, if the phenology of host plants is drastically altered, then many herbivores may be left without food. Previous research on the effects of climate change on interspecific interactions has focused on bitrophic interactions. My research expands this past emphasis to a tritrophic level: to interactions between plants (penduculate oak, Quercus robur), herbivorous insects (moths: Tischeria ekebladella, Phyllonorycter quercifoliella and P. harrisella) and parasitoids (wasps in the family Eulophidae). Since bud burst in oaks varies significantly among individuals, I also investigated how host genotype may affect the synchrony between species. My hypotheses were that higher spring-time temperatures will disrupt the synchrony between species, and that this disturbance will affect the growth and reproduction of species at higher trophic levels. I also posited that the effects will vary with the genotype of the host. I tested these hypotheses in a field experiment running from April to September 2009. To manipulate temperatures I used a greenhouse, transplanting multiple oak-specific moth and parasitoid species to oaks of different genotypes growing inside and outside of the greenhouse. During the field experiment, the temperature differed by 3.19°C between the interior and the outside of the greenhouse. As a result, the phenology of all species was advanced in the greenhouse interior as compared to ambient conditions. Interspecific synchrony was affected differently in different species pairs: Inside of the greenhouse, the synchrony between oaks and Phyllonorycter moths and between oaks and parasitoids was decreased. The synchrony between oaks and the moth T. ekebladella remained unaffected. Likewise, the synchrony between moths and parasitoids was left intact. The larvae of T. ekebladella grew bigger inside of the greenhouse than outside of it. In addition, a second generation of T. ekebladella occurred inside of the greenhouse, but not under ambient temperatures. Host plant phenology had a significant effect on associated insects: moth larvae of T. ekebladella grew bigger on oaks sprouting leaves early in the season. This increase in performance suggests that moth larvae prefer old leaves. In summary, my study shows that climate change may affect different species and different interspecific interactions in highly different ways. My study also upsets the previous view that mature oak foliage would offer food of inferior quality to moth larvae. In addition, it depicts host plant genotype as a less prominent determinant of insect performance than has been previously assumed.
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(2014)Mobile zinc is involved in the pathogenesis of several fatal neurodegenerative diseases, such as amyotrophic lateral sclerosis, Alzheimer's disease, and Parkinson's disease. Design of novel Zn(II) chelators is a promising research field in the development of new medical treatments for these diseases. However, depletion of zinc using a high affinity chelator can lead to cell death. The Zn(II) chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) can reverse some zinc related pathologies, but its high zinc affinity makes it unsuitable for use as a medical treatment. In this thesis, calculations at the density functional theory (DFT) level have been performed on TPEN and TPEN derivatives. The aim was to suggest modifications in the molecular structure which lower the zinc affinity of the chelator to a less toxic level, and which therefore could potentially lead to new medical therapies for neurodegenerative diseases. A further aim was to develop a computational protocol that is suitable for studies and in silico design of Zn(II) chelators. The results show that DFT methods, which include a correction for dispersion forces and which treat the solvent implicitly, can yield free energies for ligand exchange reactions which agree well with experimental data. The employed computational methodology is also suitable for similar studies involving other metals. The zinc affinity of TPEN can be lowered by substituting hydrogens on its pyridyl rings with electron-attracting groups. Substitution with weakly electron-donating groups can also lower the zinc affinity, provided that it results in a conformational change which stabilises the free chelator. Substitution of carbon atoms with nitrogens on the pyridyl rings also lowers the zinc affinity. The computational methodology needs improvement if one wishes to address more complicated problems, such as studies of complexation energies for chelators with varying denticities, in which solvent molecules may play a more significant role as one of the ligands.
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(2017)Because of their eusociality and diverse adaptations, ants are classic study objects in evolutionary biology. Supercolonies consisting of numerous nests have recently received much attention. Supercoloniality is an ecologically dominant and successful lifestyle, but it may be an evolutionary dead end. A supercolony has extremely many queens, and relatedness between colony members is thus very low. Inclusive fitness theory predicts selfish traits to spread in such low relatedness colonies, and if relatedness is zero, kin selection cannot act on worker traits. In such situations eusociality cannot persist. Dispersal is dangerous to daughter queens, and only a small fraction succeeds in establishing new nests. When relatedness between daughter queens and the rest of a colony is high, kin selection will, however, favor dispersal, because competition against relatives does not benefit daughter queens. But when relatedness is low, daughter queens may maximize their inclusive fitness by staying in their natal colony, although dispersal would be the collective interest of the colony. Non-dispersing by daughter queens is thus selfish behavior and is expected in low relatedness colonies. I studied dispersal between a large nest aggregation and other smaller colonies of the ant Formica pressilabris in Raseborg, Southern Finland. I found a supercolony-like dense aggregation of more than 1 300 nests on a 9 ha large abandoned field, as well as three other nest aggregations a few hundreds of meters away from the assumed supercolony. I studied dispersal between these subpopulations indirectly by estimating gene flow using microsatellite DNA markers. I extracted DNA from 285 nests and studied ten microsatellite loci. In addition to the population genetic study, I performed behavioral experiments, on one hand to determine whether supercolony workers identify intruders at all, and on the other hand to investigate whether the large nest aggregation actually is one supercolony. My results show limited gene flow between the four subpopulations. The two largest subpopulations are viscous, i.e. neighboring nests are genetically more similar to each other than to more distant nests in the same subpopulation. However, my results do not support my hypothesis that supercolony daughter queens disperse less than daughter queens from other colonies in the area. One explanation for this result is that non-dispersal of daughter queens does not show up in microsatellite studies. This might be the case if there is enough male dispersal to even out the gene flow between subpopulations. Another possible explanation is that dispersal is limited from all of the subpopulations, which all seem to be polydomous. Thirdly, it may be that even supercolony daughter queens disperse, which would be against my hypothesis. This possibility is supported by the weakness of the population structuring. These three alternatives are mutually nonexclusive and may all affect my results. In my behavioral experiments I found aggression between nests of the nest aggregation assumed to be a supercolony. Thus, it is not a uniform supercolony, as ants of a colony are by definition not aggressive towards each other. This result is surprising, and such a supercolony-like nest aggregation with aggression between its nests has not been reported earlier. If the nest aggregation actually consists of many smaller polydomous colonies, the result from my population genetic study, which is against my a priori hypothesis, would be expected. My results underline that sufficient attention should be paid to the interactions between individual nests when studying supercolonies.
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(2014)This project was about the molecular mechanisms involved in the generation of eicosanoids in human mast cells with particular emphasis on lipid bodies as a source and/or site of lipid mediator biogenesis. The cells to be used are isolated from human peripheral blood provided by Finnish Red Cross Blood Transfusion Service and collected from healthy donors. Human mast cells are found in connective tissue. They contain granules filled with histamine, heparine and proteases. Human mast cells are potent effector cells in host-defense mechanisms of innate immunity, including inflammatory diseases such as atherosclerosis. Activation of mast cells by different stimuli triggers the release of a huge range of mediators, including de-novo synthesized eicosanoids, which are highly biologically active lipid mediators. The major eicosanoid released by activated mast cells is prostanoid prostaglandin D2 (PGD2). The aim of this project was to find out whether mast cell lipid bodies are the cellular compartments of PGD2 synthesis, what are the enzymes involved in AA liberation from TGs, and whether TG-derived AA is a source for PGD2 production. The enzymes of special interest were hormone sensitive lipase (HSL) and adipose triglyceride lipase (ATGL). We were also interested about hematopoietic prostaglandin D synthase (HPGDS), the key enzyme in the production of D and J series of prostanoids. Methods used in this pro gradu work include siRNA transfections, RNA isolation, cDNA synthesis, qPCR, immunoblotting, ELISA and conventional fluorescence microscopy. Immediate increase in the amount of PGD2 released from mast cells sensitized with human IgE (1 µg/ml) and activated by polyclonal rabbit anti-human IgE (1 µg/ml) was observed. The increase was most prominent after one hour of activation, and slowly decreased to basal levels at 48 h post-activation. siRNA transfection affected the amount of enzyme DNA in mast cells and the amount of PGD2 released. HSL, ATGL and HSL+ATGL double knockdowns all reduced the amount of PGD2 released in acute (5 to 30 minutes) term activation compared to control cells. However, no significant changes were observed in the mRNA expression levels of ATGL, HSL, CGI-58, HPGDS or COX-1 under mast cell activation. The only significant changes in mRNA expression levels were observed with COX-2. However, the relative expression of HPGDS increased in IgE treated mast cells compared to control treated cells and the expression was even greater in mast cells treated with αIgE also. Both ATGL and HPGDS were recognized throughout the cytosolic area in the non-activated Ctrl cells. Although HPGDS located also in the circumference of mast cells, no clear localization of HPGDS was observed in the circumference of mast cell lipid droplets. The experiments carried out at the Wihuri Research Institute, including those presented here, have established that, in addition to phospholipids, the triglycerides present in mast cell lipid droplet core are also an important source of eicosanoids, and that also ATGL and HSL, not just cPLA, can release arachidonic acid for eicosanoid production. The ramifications of this study include the possibility that arachidonic acid release from triglycerides for the formation of eicosanoids could take an indirect or a direct route to supply precursors for cellular eicosanoid biosynthesis. The key is the pathway of AA release. In the direct pathway, AA is released from LD TGs by ATGL or HSL and this free AA is used for the generation of PGs by either COX-1 or COX-2, depending on the status of the cell. In the indirect pathway, AA is liberated from LD TGs by ATGL or HSL and then further re-esterified into phospholipids from where AA is then finally released by cPLA2 for the generation of eicosanoids.
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