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  • Niskanen, Julia (2016)
    Epidermolysis bullosa (EB) is a group of hereditary skin disorders caused by mutations in the genes that code for adhesion molecules in keratinocytes. The symptoms of the disease include blisters and erosions in the skin as well as abnormalities in the mucosal membranes, nails and tooth enamel. Depending on the causative mutation the severity of the disease ranges from mild to lethal. This master s thesis was carried out in professor Hannes Lohi s canine genetics research group. It is a case study aiming to identify the genetic cause of EB present in Central Asian shepherd dogs. Furthermore, the mode of inheritance, frequency of the mutation in the breed and the effect of the mutation on the tissue were also examined. Primary research material included blood and tissue samples from a family of Central Asian shepherd dogs. Additional samples were obtained from other Central Asian shepherd dogs as well as from dogs of closely related breeds. All dogs included in this study are owned by private persons, and participation in this study was voluntary. The research methods used in this study included both wet laboratory experiments and bioinformatic in silico procedures. The genome of one affected dog was sequenced in order to identify the gene causing EB, and data from the sequencing was filtered with multiple programs according to recessive model. The model was decided after analyzing the pedigree of the affected dogs. After finding the likely causative gene the mutation was validated in a larger cohort with Sanger sequencing. Protein expression in the tissue of affected dogs was also studied using immunofluorescence staining. As a result of this study, a new mutation causing recessive dystrophic EB was identified. The mutation is specific to Central Asian shepherd dogs. Affected dogs have a homozygous mutation in the COL7A1 gene, which codes for collagen VII α1 protein. The mutation causes a premature stop codon in the mRNA sequence, which results in abnormal protein production and separation of skin layers. The frequency of the mutation allele in the sample is approximately 18 % and more than a fourth of the dogs in the sample are carriers. Based on the pedigree analysis, the mutation is relatively new and it is only found in a small population. The disease can be prevented from becoming more common in the breed with the help of a gene test, and the test will be available in the commercial MyDogDNA gene test panel.
  • elDandashi, Rahaf (2021)
    Epigenetics is the study of changes in gene function without affecting the DNA sequence. Epigenetics studies the effects of the environment and behavior on the genome. Researchers have been able to detect several epigenetic modifications such as –DNA methylation, histone acetylation, and microRNA-associated gene silencing. Changes in the epigenome are essential for proper cell function and normal development and can also be induced by environmental factors. Stress is defined as a biological response to physiological and psychological demands which can affect cellular homeostasis. Factors such as prenatal life stress can affect gene function without directly altering the DNA nucleotide sequence. Elevated levels of stress can immobilize with the ability to impair cognitive function. There is evidence that suggests the involvement of epigenetic regulation in disorders such as addiction, depression, schizophrenia, and cognitive dysfunction. Therefore, this systematic review discusses recent findings of the role of epigenetics in prenatal exposure to stress. To achieve this, the thesis will cover different subtopics from genetics, neurobiology, and diseases, neuroscience, biological psychiatry, life sciences, medicine, behavioral brain research, biochemistry & molecular biology, as well as neuroendocrinology. Research questions are 1) Is there an association between epigenetics and prenatal stress? 2) What kind of mechanisms have been found? 3) What kind of techniques have been used in the identification of potential epigenetic mechanisms? What genes are associated with these epigenetic changes?. This study followed the "The Preferred Reporting Items for Systematic Reviews and Meta-Analyses" (PRISMA) guideline checklist. Eligibility criteria and search terms where be selected and documented to offer the widest range of articles covering the subjects of this study. A literature search was done using PubMed/Medline, Google scholar, and gray literature. The last sample comprised 59 articles. Data were extracted so that the participants, intervention, comparisons, and outcomes were included. The literature search conducted in this systematic review identified a few findings. First is that the majority of animal and human studies found a significant or moderate association between epigenetics and prenatal stress. Second, DNA methylation is the most studied epigenetic mechanism in maternal exposure to stress Third, genome-wide studies were more common in human studies than in animals and the most widely used method used is Infinium HumanMethylation450 Bead Chip. However, the common methods used in human and animal studies are most likely because of the small sample size and causation cannot be determined. Finally, NR3C1 and FKBP5 genes were the most studied in human studies where they showed the strongest association between prenatal stress and epigenetic modifications. While in animal studies, the most studied genes were Bdnf and Dnmt1 as they showed a significant methylation level after maternal prenatal stress exposure. In conclusion, maternal prenatal stress could trigger epigenetic alterations in neonates in both animals and humans. This holistic review detailed and evaluated locus-specific and studies exploring current knowledge about associations between maternal prenatal stress and epigenetic changes.
  • Matikainen, Riikka (2019)
    In this master’s theses, Finnish biology teachers’ needs for material for continuing education and educational material for upper secondary school in epigenetics were studied. Two sets of educational materials, continuing educational material for teachers and educational material for upper secondary school teaching was produced accordant with the results. Epigenetics is used to describe stable alterations in gene expression which do not consider mutations in DNA sequence. DNA methylation, histone modifications and chromatin remodelling are the main epigenetic mechanisms to affect gene expression. Epigenetic modification patterns can alter de novo, or they can be originated by some environmental factor. Epigenetic regulation was introduced as a new subject matter in National Core Curriculum for General Upper Secondary Schools 2015 in Finland. Epigenetics is a relatively new branch in biology, and as a result many teachers have not studied the subject matter at the university. Continuing educational material is needed to update their knowledge. Biology teachers’ needs for material in epigenetics were studied with a survey which was distributed to the Biology and Geography Teachers’ Union’s e-mail list subscribers. Data was analysed qualitatively and quantitatively. In order to produce continuing educational material for teachers, literature regarding teacher competence, adult education and Finnish biology teachers’ subject expertise was examined. The concept of constructivist learning, and conceptual change were applied in the production of educational material for upper secondary school teaching. Relevant scientific literature in epigenetics was gathered and used to produce both sets of materials. In the study survey, the teachers reported specific individual educational needs which were acknowledged in the production of both sets of materials, alongside literature listed earlier. The survey showed that one of 33 biology teachers had studied epigenetics at the university, 20 of 33 teachers independently and 13 of 33 hadn’t studied epigenetics at all. The extent of the teachers’ studies in epigenetics was most often elementary and the main motivation to study epigenetics independently was a desire to handle the basics. The most common resource teachers used to study epigenetics was non-scientific magazines. Among the teachers who had not studied epigenetics at all, lack of time was the most common reason mentioned. However, 14 teachers described epigenetics as an important subject matter. Three teachers reported that they lack the expertise in teaching epigenetics and three felt that textbooks don’t offer support in teaching epigenetics. Online material and expert lectures were the most common continuing education material forms requested. Regarding the content of the continuing educational material for teachers, the most common requests were that the material should include the basics of epigenetics and practical examples. The form of educational material for upper secondary school teaching was most commonly requested to be online text or educational video. Regarding the content of the educational material for upper secondary school teaching, the most common requests among teachers were a summary of the theory of epigenetics and practical examples. The continuing educational material for teachers produced in this thesis consists of an introduction part and four parts about different subjects in epigenetics. The titles of the parts are: 1) What is epigenetics? 2) Molecular mechanisms of epigenetic gene regulation 3) Epigenetic gene regulation and 4) Epigenetic inheritance. The material was designed in a way that texts 3) and 4) are possible to comprehend without studying text 2) about molecular mechanisms of epigenetic regulation. The educational material for upper secondary school teaching consists of seven parts. At the beginning there is an introduction to teachers which is followed by six separate parts for students. Each part has a “Rehearse before reading”-box which introduces students to the subject and encourages rehearsal of biological concepts and vocabulary which are necessary for comprehending each part. After each part there are questions which test students’ learning as well as encourage to apply freshly acquired knowledge about epigenetics to other biological contexts. Titles for different parts are: 1) What is epigenetics? 2) Epigenetics and nutrition 3) Epigenetics and exercise 4) Epigenetics and mental health 5) Epigenetics and tortoiseshell cats and 6) Epigenetic inheritance. The material has been designed in such a way that the parts can be taught and learned separately. References are provided at the end of both material sets. Both materials produced in this thesis meet the teachers’ requests revealed in the survey. The form of the continuing educational material for teachers is online material, which was one of the most common requests among the teachers who answered the survey. The contents of the material correspond to the teachers requests as well, since most requested contents were the basics of epigenetics and practical examples. The educational material for upper secondary school teaching was requested as online material with a summary of the theory of epigenetics and practical examples, and all these requests were met. Both materials were produced considering relevant theories on pedagogy and adult education. Results of this study cannot be applied nationally in Finland since the sample size was small. Therefore, national relevance of the materials cannot be predicted. Predictions about the impact of the materials on teachers’ and students’ understanding about epigenetics cannot be made either, since learning is an active process which requires effort from the learner. However, a strong case can be made for the produced material, because materials include relevant information and their pedagogic choices can be justified by the literature. This thesis uncovered many questions for future research. For example, the efficacy of the materials could be studied in a practical classroom situation. Other possible questions for research or study designs could be about biology teachers’ expertise and continuing education in Finland.
  • Nieminen, Martta (2013)
    The trend of energy policy in European Union as well as in international context has lately been to increase the share of renewable biofuels. The causes for this are global warming, shrinking reserves of fossil fuels and governments' aspiration for energy independence. Microalgae have shown to be a potential source of biofuels. Though cultivation of microalgae has a long history, has production for fuel yet been unprofitable. Production has become more effective as cultivation has shifted from open ponds to controlled photobioreactors but to achieve effective cultivation methods substantially more understanding on the ecophysiology of microalgae is needed. The aim of my thesis was to research the optimal light intensity and temperature of photosynthesis for three microalgae (Chlorella pyrenoidosa, Euglena gracilis and Selenastrum sp.), which are the main parameters limiting the level of photosynthesis in nutrient rich environments such as photobioreactor. The research strains were incubated in eight light intensities (0,15-250 µmol m-2 s-2) and in 5-6 temperatures (10-35 °C). Photosynthetic activity was determined with radiocarbon method which is based on the stoichiometry of photosynthesis. The purpose of radiocarbon method is to estimate how much dissolved carbon dioxide do the algae assimilate when photosynthesizing. In the method the algae are incubated in light and dark bottles where certain amount of radiocarbon (14C) has been added as a tracer. The algae fix 14C in the proportion to available 12C. 14C method has become the most common way to measure the photosynthesis of microalgae. All of the algal strains grew in 10-30 °C but C. pyrenoidosa was the only one which grew also in 35 °C. The data was analyzed by fitting them with two photosynthesis-light intensity relationship models and one photosynthesis-temperature relationship model and as a result values of essential parameters, i.e. optimal light intensity (Iopt) and temperature (Topt) for photosynthesis, could be estimated. The model which gave the best fit was chosen to describe the photosynthesis-light intensity relationship. The optimal light intensity for C. pyrenoidosa ranged between 121–242 µmol m-2 s-2 and optimal temperature was 15 °C. Corresponding values for E. gracilis were 117-161 µmol m-2 s-2 and 24,1 °C, and for Selenastrum sp. 126-175 µmol m-2 s-2 and 16,7 °C. Q10-values were also determined. With all research strains, the level of photosynthesis increased as light intensity and temperature grew until optimal values were reached. The strains tolerated higher light intensities in warmer temperatures but after reaching the optimal temperature, the level of photosynthesis did not increase any more with elevating temperature. Robust algal strains, i.e. strains, that are most adaptable in terms of light intensity and temperature, are the most prominent ones for biofuel production. From these research strains the most adaptable strain in terms of light intensity was C. pyrenoidosa and in terms of temperature Selenastrum sp. C. pyrenoidosa had superior carbon fixation rate in relation to cell size. Therefore it can be concluded that C. pyrenoidosa is the most suitable algal strains for biofuel applications of the strains assessed here.
  • Saarinen, Eeva (2014)
    Mikrolevien käyttöä biodieselin raaka-aineena on viime vuosina tutkittu paljon niiden ollessa hiilineutraali vaihtoehto fossiilista alkuperää oleville uusiutumattomille polttoaineille. Tämän lisäksi ne ovat myös ekologisesti ja eettisesti kannattava uusiutuvan energianlähde ja tukevat pyrkimystä kohti energiaomavaraisuuden saavuttamista. Mikrolevien todellista mikso- ja heterotrofiaa on tutkittu toistaiseksi vähän eikä niiden hyödyntämän hiilen määrästä ole tarjolla tutkimustietoa. Mikroleväkantojen todellisen mikso- ja heterotrofiakyvyn selvittäminen mahdollistaa kantojen kustannustehokkaamman kasvatuksen, koska mikso- ja heterotrofiaan kykenevät kannat voivat hyödyntää kasvussaan heikompia valaistusolosuhteita korvaamalla ne osittain tai täysin orgaanisilla hiilen lähteillä. Miksotrofisten leväkantojen käytöllä voidaan myös saavuttaa kasvatuksissa korkeampi solusaanto sekä solujen korkeampi lipidipitoisuus. Heterotrofisten kantojen käytöllä voidaan taas kiertää monia mikrolevien kasvatukseen liittyviä ongelmia. Toteutin tutkimukseni kolmella mikroleväkannalla: Selenastrum sp., Chlorella pyrenoidosa ja Euglena gracilis. Tein aluksi kasvatuskokeita, joissa kasvatin kaikkia leväkantoja auto-, mikso- ja heterotrofisissa olosuhteissa. Selenastrum sp. – ja Chlorella pyrenoidosa – kannan kasvatuksissa käytin epäorgaanista COMBO – kasvatusliuosta. Euglena gracilis – kannalla käytin epäorgaanista AF6 – kasvatusliuosta. Tutkin leväkantojen potentiaalista kykyä mikso- ja heterotrofiaan glukoosin ja asetaatin lisäyksen avulla. Seurasin leväkantojen kasvua solumäärän kehityksen kautta. Kasvatuskokeiden perusteella tein päätöksen ottaa kaikki käyttämäni leväkannat mukaan varsinaisiin mikso- ja heterotrofiakyvyn mittauksiin sekä käyttää ainoastaan glukoosia orgaanisen hiilen lähteenä, sillä leväkannat menestyivät heikosti asetaattikasvatuksissa. Mikso- ja heterotrofiakyvyn mittaamiseen käytin 14C-menetelmää. Kaikkien leväkantojen kyky hyödyntää asetaattia (pitoisuus 5 g l-1) oli yleisesti heikompi verrattuna niiden kykyyn hyödyntää glukoosia (pitoisuus 5 g l-1). Kasvatuskokeissa Selenastrum sp. – kannalla vaihtelu oli eri käsittelyiden välillä solumäärän kehityksen ja kasvunopeuden suhteen suurinta. Euglena gracilis – kannalla vaihtelu solusaannon kehityksen ja kasvunopeuden välillä oli pienintä kasvatusten välillä. Selenastrum sp. – kannan miksotrofisesti ottama hiilen määrä oli suurin muihin miksotrofisesti kasvatettuihin leväkantoihin verrattuna. Heterotrofisesti otetun hiilen määrä oli suurin Euglena gracilis – kannalla. Tutkimukseni merkittävin tulos on käyttämieni leväkantojen todellisen mikso- ja heterotrofiakyvyn todistaminen. Laajemmat lisätutkimukset leväkantojen todellisesta mikso- ja heterotrofiasta ovat kuitenkin tarpeen muun muassa mikroleväbiomassan käyttösovellusten kehittämisen ja käyttöönoton sekä vesiekosysteemien hiilenkierron tuntemisen kannalta.
  • Niemistö, Emilia (2019)
    Diesel, which belong to the fossil fuels, have been exposed recently because of global warming and because of decrease of known fossil fuel reserves. It has been estimated that reserves of fossil fuels will last next 40-50 years. Burning of fossil diesels will cause not only carbon dioxide emissions but also particle and metal emissions. There have been attempt to develop alternative energy sources. Growing alternative, which have came to the market, is renewable diesels, which can be produced from many sources such as reeds and biowaste. Biodiesels differ from fossil diesels by their chemical structure. Contrary to fossil diesel, completely renewable diesels do not contain aromatic compounds and they burn more purely. Diesel might end up in soil in accidents. Behaviour of fossil diesels in a ground is well known, however, the behaviour of renewable diesels is not known so well. Even though solubility of renewable diesels is similar with fossil diesels, renewable diesels may have different behavior due to different chemical structure compared with fossil diesels. Soil removal and taking the soil to be cleaned up is the most popular way to clean diesel contaminated soil resulted from diesel leaking. Alternative way to clean diesel contaminated soil is to use bioremediation where the microbes are used. In bioremediation microbes are added to the soil or activity of the microbes is boosted by improving living conditions of microbes by adding nutrients and oxygen. In this master thesis spreading of three different diesels qualities in soil and with water in laboratory were viewed. Results of the laboratory experiment were compared to the result of Katariina Lahti-Leikas’ lysimeter experiment in Jokimaa. Diesel qualities used in experiment were completely fossil EN590, partly renewable ProDiesel and completely renewable NEXBTL. Experiment were executed in soil columns in laboratory conditions where pure lysimeter sand were exposed to diesel. Biostimulation were made to half of the soil columns using urea that every diesel had natural decomposition and biostimulation. Soil columns were watered and hydrocarbons C10-C40 from collected water below the column were analysed. In addition, soil samples were collected and hydrocarbons C10-C40 were analysed. The pH of the soil and water samples were followed throught the experiment. Biostimulation decreased concentration of hydrocarbons in the water in every diesel quality. Biggest concentration were in water from NEXBTL columns which move in soil mostly in NAPL. In soil samples there was not the same kind of decrease in hydrocarbon concentration as in water samples. Diesel moved from the upper part of the soil columns to the lower part of soil columns during the experiment. The reason why biostimulation did not decreased the hydrocarbon concentration in soil is probably because the experiment period was too short. Other things to affect the result are probably wrong comparison of nutrients or lack of oxygen, which have slowed down decomposition of hydrocarbons. The results of this study are consistent with the results of Katariina Lahti-Leikas’ lysimeter experiment in Jokimaa.
  • Hämäläinen, Maria (2020)
    Haitalliset vieraslajit aiheuttavat muutoksia luonnon monimuotoisuuteen, ekosysteemien toimintaan sekä niiden tarjoamiin ekosysteemipalveluihin vaikuttamalla esimerkiksi elinympäristöihin, ravintoketjuihin ja kilpailuasetelmiin. Haitalliset vieraskasvit voivat syrjäyttää alkuperäisiä kasveja kilpaillessaan niiden kanssa tilasta ja resursseista. Lisäksi vieraskasvit voivat heikentää paikallisten lajien elinmahdollisuuksia muokkaamalla ekosysteemejä niille sopimattomiksi. Komealupiini (Lupinus polyphyllus) on yksi yleisimmistä vieraskasveista Suomessa. Tämä kansallisesti haitalliseksi vieraslajiksi luokiteltu kasvi on levinnyt jo vuosikymmenien ajan erityisesti tienpientareilla ja joutomailla. Kasvustojen ikääntyminen voi muuttaa vieraskasvin vaikutuksia elinympäristönsä alkuperäiseen kasvillisuuteen, mutta komealupiinin kohdalla ikääntymisen vaikutuksista ei ole tietoa. Tämän pro gradu -tutkielman tavoitteena oli selvittää eri-ikäisten komealupiinikasvustojen vaikutuksia alkuperäiseen kasvillisuuteen sekä yhteisötasolla että eri lajiryhmissä. Tutkin myös kasvuston ikääntymisen vaikutusta komealupiinin peittävyyteen. Kartoitin putkilokasvilajistoa Päijät-Hämeen alueella 19 tutkimuskohteessa, jotka olivat maaperältään hiekkaisia tai soraisia. Tutkimuskohteet jaettiin komealupiinikasvuston iän mukaan kolmeen luokkaan: 1) nuoret (alle 6 v vanhat), 2) keski-ikäiset (noin 10 v) ja 3) vanhat (yli 13 v). Eri-ikäisten kohteiden kasviyhteisöjä vertaamalla tutkittiin komealupiinikasvuston ikääntymistä ja sen vaikutuksia muuhun kasviyhteisöön. Komealupiinin vaikutusten yleistämiseksi esiintyneet putkilokasvilajit luokiteltiin kymmeneen ryhmään suotuisten elinympäristöjen sekä elinympäristöjen ravinteikkuuden ja kosteuden mukaan. Aineiston analysointiin käytettiin ei-metristä moniulotteista skaalausta (NMDS) sekä yleistettyjä lineaarisia sekamalleja (GLMM). Komealupiinilla ei ollut huomattavaa vaikutusta kasviyhteisöjen rakenteeseen tai lajimääriin. Komealupiini vähensi kuitenkin kuivien elinympäristöjen lajien sekä generalistien peittävyyksiä. Komealupiinin peittävyys ei vähentynyt kasvuston ikääntyessä. Komealupiini ei vaikuttanut kovin voimakkaasti elinympäristöönsä, mikä johtui todennäköisesti siitä, että komealupiinin peittävyydet olivat melko vähäisiä (keskimäärin 35 % ± SD 12). Tämän tutkimuksen perusteella komealupiinin vaikutukset tälläisessä kuivassa elinympäristössä eivät ole kovin voimakkaat, mikäli sen peittävyys on vähäinen. Luultavasti komealupiinin haitallisuus liittyykin juuri sen kykyyn muodostaa tiheitä kasvustoja. Komealupiinin torjunnassa olisikin tehokkainta keskittyä niihin alueisiin, joissa komealupiini muodostaa tiheitä kasvustoja. Toisaalta huomiota tulisi kiinnittää myös paikkoihin, joissa kasvaa etenkin uhanalaisia kuivien elinympäristöjen lajeja sekä lajeja, jotka pärjäävät heikosti lajienvälisessä kilpailussa.
  • Uimonen, Konsta (2022)
    Finnish matriculation examination in biology transitioned into a digital examination from traditional pen-and-paper examination in spring of 2018 as a part of larger process aiming to digitalize the entire matriculation examination. Tthe requirements for the exam have also changed: in addition to subject knowledge, the digital examination also requires the candidate to also be master the digital skills outlined in the Finnish high school curriculum, for example the use of image- and data processing programs. It has been suggested hat candidates from higher socioeconomic background benefit more from digital skill teaching. It has also been shown that some teachers overestimate the digital skills of the students, omitting parts of the teaching process that are integral for learning. Tthis study examines whether the matriculation examination answers requiring digital skills differ from questions that can be answered without advanced digital skills. Statistically significantly worse performance in questions requiring digital skills might indicate digital skill teaching that is insufficient compared to the skill level required in matriculation examination. Insufficient teaching may disproportionately affect those from lower socioeconomic background. The data of this study consists of 30 full-length answers from matriculation examinations of spring 2018, autumn 2018 and spring 2019, for a total of 90 full-length answers. The data was a randomized sample provided by Ylioppilastutkintolautakunta (Finnish Matriculation Examination Board). The differences were analysed with Mann-Whitney U-test and Fisher’s z-transformation. The study found that questions requiring digital skills were statistically significantly more popular than questions that didn’t require digital skills. Although the level of cognitive processing outlined by Bloom’s Taxonomy was lower in questions requiring digital skills, there was no statistically significant difference in the success (measured by attained score) between the answers. According to results, the digital proficiency of the candidates was at a sufficient level to answer to the questions.. The study also speculates on factors that could explain the observed differences in answer popularity, cognitive depth and difficulty level of the questions.
  • Bergman, Nora (2021)
    Rapid environmental changes over the last 100 years have led to substantial range shifts across taxonomic groups. Understanding what facilitates successful shifts is important for predicting ecological consequences and planning efficient conservation actions. Interestingly, the very process of range expansion can affect the success of the shift by causing genetic changes in the expanding populations. Theory predicts that without sufficient gene flow, repeated founder events and strong genetic drift can result in allele frequency gradients and loss of genetic diversity along the expansion axis. Empirical studies testing these expectations in environment-driven range shifts are still relatively scarce, and how range expansions affect genetics in highly mobile species remains unclear. In this study, I investigated the genetic consequences of a recent range expansion in a long-distance migratory passerine, the reed warbler (Acrocephalus scirpaceus). Utilizing genome-wide data from restriction site-associated DNA sequencing (RAD-seq), I studied whether the expansion was reflected in either population structure or genetic diversity of the recently established Finnish range edge population. Despite philopatry and genetic differentiation to the range core populations, principal component analysis (PCA) and a model-based Bayesian clustering approach (fineRADstructure) revealed a lack of spatial population structure along a putative colonization route. Levels of genetic diversity, based on expected heterozygosity, nucleotide diversity, and private allele count, were found to be very similar between range edge (Finland) and range core (Central Europe). The results likely indicate high levels of gene flow both within the new population and across greater spatial distances during or after the range expansion. Due to a detected sequencing batch effect, however, the exact diversity estimates must be considered preliminary. These findings suggest that species with high enough dispersal propensity may escape the predicted genetic costs of range expansions, retaining high levels of genetic variation at range margins. This study provides valuable insights for understanding range shifts in mobile taxa, and highlights the need to investigate further the traits of species that enable the preservation of evolutionary potential during range shifts.
  • Karmila, Nelli (2022)
    Schizophrenia is a debilitating psychiatric disorder associated with reduced life expectancy. The biological mechanism of schizophrenia is nebulous; however, many findings point to the central nervous system and neurons, where a reduction in dendritic spines has been indicated by previous research. The genetic findings support the involvement of synapses in the pathogenesis of schizophrenia. To study the biological properties stemming from genetics, relevant model systems and efficient methods are needed. Induced pluripotent stem cell (iPSC) technology offers a robust method for modeling the biological processes underlying schizophrenia. Somatic cells, e.g. fibroblasts, can be reprogrammed back to a pluripotent state resembling embryonic stem cells, and further differentiated into any cell type of the body, which might not be otherwise accessible. This allows establishing and characterizing neuronal cultures from patient and control cell lines, potentially revealing biological differences associated to the disease phenotype. The field of schizophrenia research has adopted iPSC technology and multiple studies have been conducted. These include assessments of synaptic density in the produced neuronal cultures, many of which reported decreased density associated with schizophrenia. In this thesis, a modified version of Nehme et al. (2018) protocol was used to differentiate iPSCs into neurons in co-cultures with human iPSC-derived astrocytes. The overarching aim was to construct an immunocytochemistry (ICC) -based assay to measure synaptic density in the produced co-cultures. First, suitable markers for characterization by ICC were tested and selected. The markers were selected to inform about neuronal identity, maturity, and synapses of the differentiated neurons. Next, the culturing conditions were optimized regarding the cell density and coating of the culturing wells. Finally, to estimate the utility of the assay, a pilot study was performed with three cell lines derived from a healthy control and a monozygotic twin pair discordant for schizophrenia. iPSCs from these cell lines were differentiated into neurons in co-cultures with astrocytes, and then characterized with ICC using selected markers and image analysis software. The synaptic density was quantified for each cell line. The performance of the assay was evaluated with analysis of variance (ANOVA) and restricted maximum likelihood model (RELM). An assay to quantify synaptic structures in mature neurons was established. The average synaptic density for all cell lines was approximately 1 synapse per 100μm of neurite. Analysis of the data produced with the assay revealed a notable batch effect and technical variation. This suggests that further optimization is needed to reduce variance from undesired sources. The pilot data suggests that the differences in synaptic density between cases and controls may be modest, further highlighting the need for minimizing noise in the assay to improve signal to noise ratio. However, indicated by power analysis, large sample sizes are needed to identify meaningful differences between cases and controls. In light of these results, more attention should be drawn to the methodology in the field of iPSC-based studies, as the principals of the assay constructed here were similar to other synaptic assays used in previous publications.
  • Piki, Emilia (2021)
    Ovarian cancers (OCs) are gynecological malignancies that cause the most gynecological cancer related deaths due to asymptomatic early-stage development and late diagnosis. The treatment of OC has not improved significantly during the last decades, and challenges are often caused by chemoresistance and the heterogeneity of cancer cell populations. Therefore, there is an urgent need to improve OC treatment outcome and implement new targeted therapies that could address the subtype specific characteristics. The most common type of OC is epithelial ovarian cancer (EOC), that can be further divided into five subtypes with distinct molecular and histological characteristics. High-grade serous subtype represents majority of cases with up to 75% of EOC patients, while other subtypes such as low-grade serous, mucinous, clear cell and endometrioid OC being less common. Considerable progress has been made in cancer treatment via precision oncology, in which individual cancer biology and tumor molecular features are investigated and used to improve treatment decisions. For this purpose, the development of patient-derived cancer cells (PDCs) offers a good opportunity to study cancer biology in vitro and to build models for preclinical molecular profiling and functional testing. PDCs can be used to establish 2D and 3D models, and most recently, wide interest has been focused on patient-derived organoids (PDOs), that offer a better model of tumor and its microenvironment, while allowing long-term culture, cryopreservation, modification and high-throughput opportunities. In this study, the aim was to establish PDO cultures using tumor cells from low-grade serous OC patients for molecular profiling and functional drug testing. PDOs were generated from both fresh and frozen tumor tissue or ascitic samples resulting to successful development of long-term PDOs from three of the five models. In order to identify optimal culturing conditions for low-grade serous OC PDOs, two previously unpublished growth mediums were tested in parallel. The more complex of the mediums showed slightly better PDO growth in general. The immunohistochemistry staining with pan-cytokeratin and PAX8 was used to confirm the epithelial and ovarian origin of PDOs. In addition, cancer panel sequencing was performed to identify mutation profiles. Importantly, the small-scale drug testing, which was performed using conventional chemotherapeutics cisplatin and paclitaxel and targeted drugs gedatolisib and trametinib, showed sample-specific responses. In conclusion, the results from this project show that PDOs are good models for ex vivo precision medicine functional studies. Importantly, we managed to establish PDOs from frozen tumor cells, suggesting that PDOs could be initiated from living biobank samples. However, the challenges related to culturing of PDOs for functional assays included slower growth rate compared to 2D cancer cell cultures and technical challenges related to Matrigel, limiting the possibilities of high-throughput drug testing. By improving these factors, PDOs will offer an efficient 3D model for preclinical use.
  • Lyytikäinen, Veera (2020)
    It is widely acknowledged that previous efforts to communicate the severity and rate of climate change have failed. Science communication has for decades relied on the presumption that more information leads to more informed decisions, thus so far, the scientific consensus about human-caused climate change has not resulted in required changes in behaviour. Previous communication efforts have, for the most part, attributed inaction to the lack of information, but in doing so, have excluded many social and psychological elements of communication. Although raising the level of awareness about climate change has been successful, climate change remains to be perceived to be a distal threat. Recently, more sophisticated approaches have been developed to meaningfully communicate climate change, drawing attention to the framing of the communication. In this study, a new approach to science-based environmental communication is evaluated. The case study seeks to address how immersive Virtual Reality (VR) can be used as a tool in science-based environmental communication for policymakers in a locally relevant context. Via immersive VR, information about forests’ role in climate change is mediated to forest policymakers. In the science communication, climate change is framed as an experiential, local, and present risk, promoting a problem definition that focuses on the climate effects of forest utilisation. I evaluate the success of the science-based environmental communication by measuring participants’ personal responses. I focus on measuring enjoyment, interest, trust, and usability. The study participants are members of the Parliament of Finland and governmental officials from the Ministry of Agriculture and Forestry; and the Ministry of the Environment. The study material consists of feedback forms from participants (N=65) and interviews of the key actors (N=7). To consider the historical background and many conflicting interests in Finnish forest and climate politics, I focus on the comparison between the natural resource position and the environmental position. The results of this study offer compelling evidence for how differently policymakers representing these two positions perceive the usage of immersive VR in science-based environmental communication. The environmental position indicated significantly higher levels of success on all measured components. Considering that the science communication framed forest utilisation as an environmental issue, it is not surprising that participants holding the environmental position perceived the science communication to be more enjoyable, interesting, trustworthy, and usable. Accordingly, the study results provide additional support for the idea of Finnish forest policy as a polarised field of policy with two main positions. With the means of immersive VR, I was able to induce strong personal responses to the science communication. Participants holding the natural resource position were more likely to challenge the legitimacy of the information and the use of VR in science communication than participants holding the environmental position. The results point to the likelihood that communicating climate change via immersive VR can induce strong negative emotions in the participants, but when the communication is comparable with the policymaker's policy preferences, they respond more positively. The study results also suggest, that to communicate climate change more meaningfully, immersive VR should be further explored as a supplementary tool in science communication.
  • Lehtonen, Jussi (2006)
    The Seychelles magpie robin Copsychus sechellarum is among the most threatened of the twelve avian species endemic to Seychelles. Originally relatively abundant and widespread in Seychelles, its population has severely diminished, mainly on account of introduced predators and habitat destruction. The lowest count was recorded in 1965, when the number of birds was between eight and fifteen, all confined to just one island. In 1990 a successful recovery program was implemented, and the population has now recovered to approximately 150 birds on four islands. Translocations to new islands are a major part of the recovery program. So far Seychelles magpie robins have been translocated to three islands, and three more target islands still need to be chosen if the long-term goals of the project are to be reached. Determining habitat suitability at the translocation target area is essential before attempting a translocation. Cockroaches are a major part of the bird's diet. Cockroach abundance has been shown to be a good indicator of magpie robin habitat quality, but it is very laborious and time consuming to measure. The objective of this study is to build a model that can be used to predict cockroach abundance, and thereby Seychelles magpie robin habitat quality, with relatively simple and inexpensive measurements. The data for this project was collected on Cousin, Cousine and Aride islands in Seychelles during October and November 2004. Several potential predictors of cockroach abundance were measured using simple and low-cost methods. The measured variables were Seychelles skink Mabuya sechellensis abundance, altitude from sea level, slope gradient, rock type, canopy cover, shrub cover, litter depth, soil depth, the relative canopy cover of different tree species, and percentage of ground covered by litter, rock, grass, woody vegetation and bare ground. These variables were measured in several 25 × 25 m plots on each island. Cockroach abundance had been measured in the same plots previously. Several multiple linear regression models were built using these variables, with cockroach abundance as the dependent variable. The Akaike Information Criterion was used to rank the models and to find the most important explanatory variables. Stepwise regression was used as a secondary method in order to see if two different methods point to the same result. Both methods indicated that slope gradient and granitic rock type are the two most important predictors of those tested. These are features that are invariably associated with granitic terrain, and both are positively correlated with cockroach abundance. A simple comparison of average cockroach abundance between granitic and coralline rock types also showed a significant difference, with granitic terrain providing higher cockroach abundance. The islands in Seychelles can be divided into two main categories: granitic and coralline islands. Steep slopes and granitic rock are both features associated with granitic terrain and granitic islands only. Therefore, based on the results of this study, granitic islands are likely to provide higher cockroach abundance and thus better Seychelles magpie robin habitat quality. Hence it is recommended that Seychelles magpie robin translocation efforts are focused preferentially on granitic islands.
  • Suonpää, Pinja (2022)
    Each year many new-borns are at risk for long-term developmental deficits due to adverse perinatal events. Early gross motor abilities have been shown to link with cognitive development and studying infant motor behaviour may provide means to assess global neurodevelopment. This thesis aims to explore a potential association between early gross motor abilities recorded at infancy with a multi-sensor wearable jumpsuit MAIJU and later neurocognitive development assessed at two years of age. The study sample (N=26) consisted of healthy full-term infants and those with prematurity or perinatal asphyxia. Spontaneous motor activity was recorded at home with the jumpsuit. Machine learning methods were used to quantitate the time infants spent in different postures and estimate the maturity of their motor abilities, which were compared to cognitive development at two years of age with correlational- and regression analyses. There was a positive trend between early motor abilities and later cognitive development. Specifically, standing posture explained the association, such that infants who spent more time standing had better cognitive abilities at two years of age. Standing may support cognitive development by increasing opportunities for visual and manual exploration and learning. Shared neuronal circuitries for motor and cognitive functions and faster neuronal maturation may also underlie the association. The current study supports the creation of future studies with larger sample sizes to establish the potential for the use of postural and movement information obtained from wearable jumpsuit MAIJU to assess the variability of neurocognitive development of at risk and typical infants with potential goal to identify future cognitive deficits at early stage.
  • Seppälä, Sini (2019)
    Not much attention is paid on the conservation of invertebrates despite their importance to the ecosystems in general and their benefits and ecosystem services to us, humans. This study is part of a project aiming to start the Sampled Red List Index (SRLI) for spiders. The IUCN Red List Index (RLI) is used for measuring the overall extinction risk of groups of species and the sampled approach is a way to evaluate the trajectory towards extinction of megadiverse groups without the need to assess every species of the whole group of interest. A random sample of 200 spider species were selected from the global checklist and assessed according to IUCN Red List Categories and Criteria. Given the availability of data, I was able to calculate the extent of occurrence and area of occupancy for 80 species, of which 70 species had an EOO above 20000 km² and 75 species an AOO above 2000 km², the thresholds below which species can be considered threatened. The trends in EOO and AOO were assumed to be stable for most species (49 species) given the inexistence of monitoring data for any taxon. Evidence of decline was found for only 10 species, usually inferred from habitat loss. Habitat data was collected for 118 species. The most common habitat type was forest (73 species), followed by grasslands (24 species) and artificial habitats (22 species). For 44 species the habitat trend was inferred to be stable, only declining, according to available knowledge, for 14 species and increasing for one species. For the remaining 141 species the habitat trend could not be inferred and was thus assumed to be unknown. The most commonly mentioned threat types were agriculture (11 species), fires (7 species) and logging (6 species). For 39 species there were no known threats and for the rest of the 132 species the threats were unknown. Conservation actions in place were observed for 104 species, most commonly site and area protection (100 species) and resource and habitat protection (88 species). Conservation actions such as education and awareness (8 species), resource and habitat protection (7 species) and site and area management (6 species) were to take into consideration. All the 200 species were estimated to be in need of further basic research especially on threats (143 species and distribution (140 species), but also on life history and ecology (135 species). Due to several knowledge shortfalls, including the Wallacean (distribution of species), Prestonian (population trends) and Hutchinsonian (response to environmental change), no threat category could be reached for the vast majority of the species. The results show that an IUCN category could be reached for only 59 species, of which 55 were assessed as Least Concern and a threatened category was reached for only 4 species (t as Critically Endangered and one as Vulnerable). The baseline SRLI at this first point in time was 0.95 (in a 0-1 scale, where 0 means all species are extinct and 1 for all species are Least Concern). We hypothesize however that among the 141 Data Deficient species there should be a higher proportion of threatened species than among the 59 evaluated. This would be due to two reasons. First, the scarcity of information on many species might in part be due to their rarity. Second, widespread species were often the only for which an assessment could be reached, creating a bias in the dataset towards a large base SRLI value. The strategy currently imposed by IUCN is therefore clearly inadequate for taxa with scarce information, which represent the vast majority of species. I propose the future use of a different, non-random, approach to the selection of species in the SRLI and its adoption for other taxa which represent in fact most extant and threatened species.
  • Maran, Suveda Vignesh (2019)
    Extracellular vesicles (EVs) are a diverse set of cell-derived membranous vesicles which have exosomes or small EVs (40 -200nm) ,large EVs (>200 nm) and larger apoptotic bodies/oncosomes (>1000 nm). They are present in most biological fluids, such as blood, sweat, and urine etc. EVs are expressed by almost all cells.Recent evidence suggests that EVs are a form of cellular communication, resulting in an intensive investigation of their role in different biological processes and also due to their biomarker potential. Ultracentrifugation (UC) is the standard method used for the isolation of EVs from different biological fluids. UC has some limitations related to laborious operation and scalability issues. Hence, there is a need for new methods for the isolation of EVs.Nanocellulose(NC) is a biopolymer obtained from wood biomass and has the ability to be functionalised with various chemical groups on its surface. The chemical groups alter the surface properties of NC for usability in different applications. Currently, It finds extensive use in different biomedical applications due to its unique physicochemical properties. The aim of the thesis is to evaluate the use of functionalised nanocellulose (FNC) as a novel alternative compared to the standard method of ultracentrifugation in the isolation of EVs from conditioned cell culture media. Cancer cell lines and mesenchymal stromal cells (MSCs) were grown in EV-depleted cell culture media after expansion in cell culture media. The EV-depleted media was collected from the cells after 48 hours and further processed by filtration to obtain the necessary conditioned cell culture media for EV isolation. Extracellular vesicles were isolated from the conditioned cell culture media by using UC and FNC. The isolated EVs from both methods were characterised and evaluated based on their different parameters using Bicinchoninic Acid (BCA) assay, Transmission Electron Microscopy (TEM), Nanoparticle Tracking Analysis (NTA), and Western Blot. The EVs isolated by FNC were smaller in size with few vesicle-like structures around 20-40 nm compared to EVs isolated by UC 60-200nm. Most EVs isolated by UC had a higher particle-to-protein ratio which indicated more purity and reduced levels of non-vesicular contaminants in the EV isolations. The EVs isolated by FNC for many samples had a low particle-to-protein ratio indicating potentially higher levels of non-vesicular contaminants being present. Hsp70, a surface marker of EVs and soluble stress protein was observed in EVs obtained from a primary MSC and another cancer cell line among all the isolation methods. No contamination from Endoplasmic reticulum(ER) components was observed in any of the samples. The EVs isolated by FNC could not be analysed more thoroughly by some of the characterisation methods due to issues such as lack of standardised sample preparation protocols, possible interference from isolation components and sample variations. The isolation method will really benefit from further optimisation of sample preparation steps and the adoption of alternative characterisation methods for evaluation. A narrower research question with focus on FNC-EDA isolation of EVs from HOS-143B and AT-MSC will be an excellent starting point for more detailed investigations.The availability of different surface modifications, faster processing and more research on the isolation mechanism will make this novel method a great alternative to UC for isolation of EVs.
  • Järvinen, Riikka (2022)
    Urban greenspaces are valuable resources for humans and are an integral part of the urban landscape. They are also popular sites for various social and cultural activities, such as dog walking. Dog ownership rates are increasing globally at a rapid rate, hence increasing the dog populations within urban cities. The increasing number of dogs could potentially have negative consequences to the quality of urban parks. Furthermore, there is a lack of research on the impacts of dogs on urban environments. The aim of this study is to gain a better understanding of the impacts of dogs on path-side soils in urban greenspaces. Soil samples were collected from both Helsinki and Lahti. The impact of dog urine was studied by comparing path-side soil samples between areas with high dog activity (i.e., parks) and similar areas with low to no dog activity (i.e., cemeteries). Soil samples were collected three times during the growing season in 2021 at three different treatments: close by 1) an evergreen tree, 2) a lamppost, and 3) a lawn area. The samples were analyzed for soil pH, electrical conductivity, organic matter, soil moisture, NO3--N, and NH4+-N. Net N mineralization and net nitrification rates were also calculated. The results indicated that dog urine is a significant contributor to the input of nitrogen in urban parks. Soil NO3-N and NH4+-N concentrations were significantly higher in parks than in cemeteries and exhibited clear seasonal trends. Soil pH was generally lower and electrical conductivity higher in parks than in cemeteries. Poles and trees exhibited the greatest concentrations, suggesting that the impact of dog urine is highly localized around path-side objects. None of the treatments in cemeteries showed little to no changes throughout the growing season. Lastly, the net nitrification rates were significantly higher in parks during spring and summer. In contrast, net mineralization rates were generally higher in cemeteries than parks. These results confirm that urban planners need to take into consideration the impact of dogs to mitigate any potential negative consequences to the quality of urban parks.
  • Järvi, Vilja (2019)
    The insular cortex has been implicated in the neurocircuitry underlying alcohol addiction. The role of the insular cortex and its projections in regulating ethanol intake in AA (Alko-Alcohol) rats has been studied using chemogenetic tools. Chemogenetic activation of the anterior agranular insula (aAI) in AA rats through excitatory DREADDs expressed in the aAI has been found to decrease ethanol consumption. The aAI projects to the central nucleus of the amygdala (CeA), another brain region involved in the development of addiction, particularly in the withdrawal/negative affect stage. In the current study, we sought to further investigate the role of the aAI and the CeA in regulating voluntary ethanol consumption in AA rats. First, we characterized the efferent projections of the aAI in AA rats by chemogenetically activating the aAI with DREADDs and then measuring c-Fos expression in various regions of interest throughout the brain. Next, we investigated the role of the aAI --> CeA projection in ethanol intake by chemogenetically activating or inhibiting the aAI --> CeA projection using the dual viral Cre-dependent DREADD approach. We examined the effects of this manipulation on voluntary ethanol consumption in AA rats in a two-bottle choice paradigm. Finally, we examined the roles of CeA D1Rs (dopamine receptors) and 5-HT2ARs (serotonin receptors) in regulating ethanol intake by examining the effects of pharmacological agonism or antagonism of these receptors on voluntary ethanol consumption in AA rats. Our results from the first experiment reveal significant activation of brain regions including the posterior agranular insula, the mediodorsal nucleus of the thalamus, and the posterior piriform cortex following chemogenetic activation of the aAI. The projections from the aAI to these regions are potentially important in the aAI circuitry in AA rats and are therefore of interest in future studies on the role of aAI circuitry in ethanol intake. In the second experiment, we found no significant effects of aAI --> CeA projection activation or inhibition on ethanol consumption in AA rats, indicating that this projection may not be a key component in regulating ethanol intake in these rats. Finally, we found no significant effects of pharmacological D1R antagonism, 5-HT2AR antagonism, or 5-HT2AR agonism in the CeA on ethanol intake in AA rats, although there was a non-significant trend towards a dose-dependent decrease in ethanol consumption with increasing dose of the D1R antagonist. Our results reveal new neural projections that should be investigated in future research on the role of the aAI in regulating ethanol intake. Studies on the neurobiology underlying alcoholism may reveal new pharmacological or anatomical targets for treatments of alcoholism in humans.
  • Partti, Edvard (2018)
    Kaurapohjaiset elintarvikkeet ovat terveellisiä. Monet niiden terveyshyödyt johtuvat kauran liukoisen ravintokuidun suuresta β-glukaanipitoisuudesta. β-glukaanin terveysvaikutukset ovat riippuvaisia sen molekyylipainosta ja viskositeetista. Viskositeetilla on myös muuta merkitystä kaurapohjaisissa elintarvikkeissa kuten kaurajugurteissa ja kauramaidoissa. Aiemmassa Folafibre-tutkimusprojektissa oli tutkittu kaurakuidun folaattipitoisuuden (B9 vitamiini) kasvattamista fermentoimalla sitä eri mikrobeilla. Hyvin folaattia tuottaneet mikrobit myös alensivat kaurakuituvalmisteen viskositeettia, ja erittivät glykosyylihydrolaaseja ja/tai proteaasia. Oli kuitenkin jäänyt epäselväksi, olivatko entsyymit ainut syy viskositeetin alenemiseen, ja kuinka paljon kullakin niistä oli vaikutusta viskositeettiin. Lisäksi tämän alan kirjallisuudessa on pidetty epäselvänä, onko esim. tärkkelyksen ja β-glukaanin välillä jotain interaktioita jotka mm. nostavat niiden viskositeettia yli yksittäisten polymeerien viskositeetin summan. Näitä voitaisiin selvittää puhdistamalla viskositeettia alentaneista mikrobikannoista kyseiset entsyymit ja tutkimalla niiden vaikutusta kaurakuidun viskositeettiin. Yhdeksi mikrobiksi valittiin Exiguobacterium sp. RB3 kanta, koska em. syiden lisäksi tässä bakteerisuvussa esiintyy psykrofiilejä ja mikrobeja jotka voivat kasvaa korkeassa pH:ssa, jolloin sen erittämät entsyymit saattaisivat olla aktiivisia matalissa lämpötiloissa ja korkeassa pH:ssa, ja olla siten teollisesti kiinnostavia. Toiseksi mikrobiksi valittiin Bacillus sp. ABM5119, koska sitä oli käytetty monissa Folafibre-projektin tutkimuksissa. Työn tavoitteena oli puhdistaa Bacillus sp. ABM5119:n endo-β-1,4-glukanaasi, ja Exiguobacterium sp. RB3:n α-amylaasi ainakin siinä määrin, että ne ovat muista endoaktiivisista glykosyylihydrolaaseista ja proteaaseista puhtaita, ja sitten mitata puhdistettuja entsyymejä ja proteaasia käyttäen niiden vaikutus yhdessä ja erikseen keitetyn kaurakuituvalmisteen viskositeettiin. Lisäksi tavoitteena oli karakterisoida Exiguobacterium sp. RB3 amylaasin olennaisimmat biokemialliset ominaisuudet, sekvensoida sen geenin amylaasia koodaava alue, ja selittää sen ominaisuuksia myös sekvenssistä johdettavissa olevan tiedon perusteella. Keitetyn kaurakuitupreparaatin viskositeettia alentavat eniten α-amylaasi ja endo-β-1,4-glukanaasi. Proteaasi ei vaikuta kaurakuitupreparaatin viskositeettiin, kun kuitupreparaatti on keitetty. Endo-β-1,4-glukanaasi alentaa viskositeettia yhtä hyvin kuin β-1,3-1,4-glukanaasi. Synergiaa viskositeetin alentamisessa α-amylaasilla ja endo-β-1,4-glukanaasilla ei havaittu, mutta havaittiin että β-glukaani saattaa estää tärkkelyksen retrogradaatiota. Hyvin pienetkin entsyymiaktiivisuudet vaikuttavat viskositeettiin. Jos halutaan β-glukaanin viskositeetin ja siten terveysvaikutusten säilyvän, täytyy esim. kauramaidon valmistuksessa käytettävien entsyymivalmisteiden olla β-glukanaaseista hyvin puhtaita. Mikrobifermentaatioissa tapahtuvat viskositeetin alenemat johtuvat lähinnä mikrobien erittämistä glykosyylihydrolaaseista, ei niinkään esim. niiden metabolian sivutuotteista kuten happiradikaaleista. Exiguobacterium sp. RB3 α-amylaasi on rakenteeltaan Bacillus licheniformis α-amylaasin kaltainen glykosyylihydrolaasiryhmän 13 entsyymi. Se sitoo rakenteeseensa kolme kalsiumiatomia, ja kalsiumpitoisuus vaikuttaa sen aktiivisuuteen. Se on aktiivisimmillan pH alueella 5,0 – 7,5. Se sietää detergenttejä, toisin kuin eräs aiemmin karakterisoitu Exiguobacterium-α-amylaasi. RB3 α-amylaasin turnover number oli korkea, 29000 1/s. Exiguobacterium-suvun psykrofiilisessä haarassa esiintyy kahta eri α-amylaasia, joista yksi on tässä karakterisoitu, ja toinen on selvästi erilainen rakenteeltaan ja biokemiallisilta ominaisuuksiltaan.