5,661 research outputs found

    Studying the interplay between ageing and Parkinson's disease using the zebrafish model

    Get PDF
    Parkinson’s disease (PD) is a neurodegenerative disorder characterised by the loss of dopaminergic neurons in the substantia nigra. Ageing is the major risk factor for developing PD but the interplay between ageing and PD remains elusive. To investigate the effect of ageing on PD-relevant pathological mechanisms, zebrafish mutant lines harbouring mutations in ageing-associated genes (klotho-/-, sirt1-/-, satb1a-/-, satb1b-/- and satb1a-/-;satb1b-/-) were generated, using CRISPR/Cas9 gene editing. Likewise, a chemical model for SIRT1 deficiency was utilised. klotho-/- zebrafish displayed an accelerated ageing phenotype at 3mpf and reduced survival to 6mpf. Dopaminergic neuron number, MPP+ susceptibility and microglial number were unaffected in klotho-/- larvae. NAD+ levels were decreased in 6mpf klotho-/- brains. However, ATP levels and DNA damage were unaffected. sirt1-/- zebrafish did not display a phenotype through adulthood. il-1β and il-6 were not upregulated in sirt1-/- larvae, and chemical inhibition of sirt1 did not increase microglial number. cdkn1a, il-1β and il-6 were not upregulated in satb1a-/- and satb1b-/- larvae. Dopaminergic neuron number and MPP+ susceptibility were unaffected in satb1a-/- larvae. However, satb1b-/- larvae demonstrated a moderate decrease in dopaminergic neuron number but equal susceptibility to MPP+ as satb1b+/+ larvae. Adult satb1a-/- but not adult satb1b-/- zebrafish were emaciated. satb1a-/-;satb1b-/- zebrafish did not display a phenotype through adulthood. Transgenic zebrafish expressing human wildtype α-Synuclein (Tg(eno2:hsa.SNCA-ires-EGFP)) were crossed with klotho-/- and sirt1-/- zebrafish, and treated with a sirt1-specific inhibitor. Neither genetic cross affected survival. The klotho mutation did not increase microglial number in Tg(eno2:hsa.SNCA-ires-EGFP) larvae. Likewise, sirt1 inhibition did not induce motor impairment or cell death in Tg(eno2:hsa.SNCA-ires-EGFP) larvae. In conclusion, the suitability of zebrafish for studying ageing remains elusive, as only 1 ageing-associated mutant line displayed accelerated ageing. However, zebrafish remain an effective model for studying PD-relevant pathological mechanisms due to the availability of CRISPR/Cas9 gene editing, neuropathological and neurobehavioral tools

    Beam scanning by liquid-crystal biasing in a modified SIW structure

    Get PDF
    A fixed-frequency beam-scanning 1D antenna based on Liquid Crystals (LCs) is designed for application in 2D scanning with lateral alignment. The 2D array environment imposes full decoupling of adjacent 1D antennas, which often conflicts with the LC requirement of DC biasing: the proposed design accommodates both. The LC medium is placed inside a Substrate Integrated Waveguide (SIW) modified to work as a Groove Gap Waveguide, with radiating slots etched on the upper broad wall, that radiates as a Leaky-Wave Antenna (LWA). This allows effective application of the DC bias voltage needed for tuning the LCs. At the same time, the RF field remains laterally confined, enabling the possibility to lay several antennas in parallel and achieve 2D beam scanning. The design is validated by simulation employing the actual properties of a commercial LC medium

    KYT2022 Finnish Research Programme on Nuclear Waste Management 2019–2022 : Final Report

    Get PDF
    KYT2022 (Finnish Research Programme on Nuclear Waste Management 2019–2022), organised by the Ministry of Economic Affairs and Employment, was a national research programme with the objective to ensure that the authorities have sufficient levels of nuclear expertise and preparedness that are needed for safety of nuclear waste management. The starting point for public research programs on nuclear safety is that they create the conditions for maintaining the knowledge required for the continued safe and economic use of nuclear energy, developing new know-how and participating in international collaboration. The content of the KYT2022 research programme was composed of nationally important research topics, which are the safety, feasibility and acceptability of nuclear waste management. KYT2022 research programme also functioned as a discussion and information-sharing forum for the authorities, those responsible for nuclear waste management and the research organizations, which helped to make use of the limited research resources. The programme aimed to develop national research infrastructure, ensure the continuing availability of expertise, produce high-level scientific research and increase general knowledge of nuclear waste management

    Early Neanderthal social and behavioural complexity during the Purfleet Interglacial: handaxes in the latest Lower Palaeolithic.

    Get PDF
    Only a handful of ‘flagship’ sites from the Purfleet Interglacial (Marine Isotope Stage 9, c. 350-290,000 years ago) have been properly examined, but the archaeological succession at the proposed type-site at Purfleet suggests a period of complexity and transition, with three techno-cultural groups represented in Britain. The first was a simple toolkit lacking handaxes (the Clactonian), and the last a more sophisticated technology presaging the coming Middle Palaeolithic (simple prepared core or proto-Levallois technology). Sandwiched between were Acheulean groups, whose handaxes comprise the great majority of the extant archaeological record of the period – these are the focus of this study. It has previously been suggested that some features of the Acheulean in the Purfleet Interglacial were chronologically restricted, particularly the co-occurrence of ficrons and cleavers. These distinctive forms may have exceeded pure functionality and were perhaps imbued with a deeper social and cultural meaning. This study supports both the previously suggested preference for narrow, pointed morphologies, and the chronologically restricted pairing of ficrons and cleavers. By drawing on a wide spatial and temporal range of sites these patterns could be identified beyond the handful of ‘flagship’ sites previously studied. Hypertrophic ‘giants’ have now also been identified as a chronologically restricted form. Greater metrical variability was found than had been anticipated, leading to the creation of two new sub-groups (IA and IB) which are tentatively suggested to represent spatial and perhaps temporal patterning. The picture in the far west of Britain remains unclear, but the possibility of different Acheulean groups operating in the Solent area, and a late survival of the Acheulean, are both suggested. Handaxes with backing and macroscopic asymmetry may represent prehensile or ergonomic considerations not commonly found on handaxes from earlier interglacial periods. It is argued that these forms anticipate similar developments in the Late Middle Palaeolithic in an example of convergent evolution

    Development of in-vitro in-silico technologies for modelling and analysis of haematological malignancies

    Get PDF
    Worldwide, haematological malignancies are responsible for roughly 6% of all the cancer-related deaths. Leukaemias are one of the most severe types of cancer, as only about 40% of the patients have an overall survival of 10 years or more. Myelodysplastic Syndrome (MDS), a pre-leukaemic condition, is a blood disorder characterized by the presence of dysplastic, irregular, immature cells, or blasts, in the peripheral blood (PB) and in the bone marrow (BM), as well as multi-lineage cytopenias. We have created a detailed, lineage-specific, high-fidelity in-silico erythroid model that incorporates known biological stimuli (cytokines and hormones) and a competing diseased haematopoietic population, correctly capturing crucial biological checkpoints (EPO-dependent CFU-E differentiation) and replicating the in-vivo erythroid differentiation dynamics. In parallel, we have also proposed a long-term, cytokine-free 3D cell culture system for primary MDS cells, which was firstly optimized using easily-accessible healthy controls. This system enabled long-term (24-day) maintenance in culture with high (>75%) cell viability, promoting spontaneous expansion of erythroid phenotypes (CD71+/CD235a+) without the addition of any exogenous cytokines. Lastly, we have proposed a novel in-vitro in-silico framework using GC-MS metabolomics for the metabolic profiling of BM and PB plasma, aiming not only to discretize between haematological conditions but also to sub-classify MDS patients, potentially based on candidate biomarkers. Unsupervised multivariate statistical analysis showed clear intra- and inter-disease separation of samples of 5 distinct haematological malignancies, demonstrating the potential of this approach for disease characterization. The work herein presented paves the way for the development of in-vitro in-silico technologies to better, characterize, diagnose, model and target haematological malignancies such as MDS and AML.Open Acces

    Varastest embrüotest pärit ekstratsellulaarsed vesiikulid: potentsiaal embrüokvaliteedi markeritena ja roll embrüo-emaka suhtluses

    Get PDF
    Väitekirja elektrooniline versioon ei sisalda publikatsiooneViljatus on globaalne rahvatervise probleem, mis mõjutab miljoneid inimesi. Abistav reproduktiivtehnoloogia, sealhulgas in vitro viljastamine, on aidanud mitmeid viljatuid inimesi. Küll on sellel metoodikal üheks kitsaskohaks implantatsiooni ebaõnnestumine isegi morfoloogiliselt parimate embrüotega. Seetõttu toimuvad jätkuvalt uuringud tuvastamaks paremaid meetodeid, mis hindavad embrüo kvaliteeti ja ennustavad siirdamise edukust, olles peamiselt embrüokasvusöötme baasil. Rakuvälised ehk ekstratsellulaarsed vesiikulid (EV) on membraaniga ümbritsetud nanoosakesed, mida toodavad peaaegu kõik rakutüübid erinevates füsioloogilistes ja patoloogilistes konditsioonides. Nende kaudu toimub rakuvaheline suhtlus. Mitmed uuringud, eriti vähi korral, on uurinud EVde potentsiaali biomarkerina ja ravimkandursüsteemina. Antud doktoritöö uuris implantatsiooni-eelse perioodi embrüost vabanenud EVde potentsiaali embrüokvaliteedi markerina ja embrüo-emaka suhtluse vahendajana. Katsed viidi läbi kasutades veise-embrüoid ja inimrakukultuuride põhiseid eksperimentaalmudeleid. Esimene uuring tõestas, et individuaalselt kasvatatud implantatsiooni-eelse perioodi veise-embrüod eritavad EVsid kasvusöötmesse ning nende kontsentratsiooni- ja suurusprofiil sõltub embrüo kvaliteedist ja arengustaadiumist. Järgnevalt katsetati munajuharakkudel implantatsiooni-eelse perioodi embrüost pärit EVde funktsionaalsust. Katse käigus selgus, et EVd kõrge kvaliteediga embrüotest muutsid munajuharakkude geeniekspressiooni, mida aga ei teinud halva kvaliteediga embrüote EVd. Suurenenud ekspressiooniga geenide hulgas olid mitmed interferoon-τ raja interferooni stimuleerivad geenid. Interferoon-τ peetakse mäletsejaliste tiinuse tuvastusmolekuliks. See leid viitab, et munajuha tunneb ära kvaliteetse embrüo. Viimaseks uuriti embrüo EVde funktsionaalsuse spetsiifilisust. Leiti, et endomeetrium reageerib vaid embrüo päritolu EVdele. Uuringute käigus tuvastati embrüost vabanenud EVde potentsiaal ja spetsiifilisus embrüokvaliteedi biomarkerina.Infertility is a global public health problem that affects millions of people in their reproductive life. Assisted reproductive technologies (ARTs) such as in-vitro fertilization have enabled many patients to overcome this issue. However, a bottleneck in ART success is the implantation failure even after the transfer of morphologically best embryos. Hence, investigations continue to identify better or complementary methods of assessing embryo quality and predicting transfer success, mainly based on the embryo culture media. Extracellular vesicles (EVs) are membrane-bound nanoparticles released by almost all types of cells under different physiological and pathological conditions. They mediate intercellular communication. Many studies, especially related to cancer, have investigated EVs' potential as biomarkers and therapeutic drug delivery systems. This project investigated preimplantation embryo-derived extracellular vesicles as a potential embryo quality marker and a mediator of embryo-maternal communication. Experiments were performed using bovine embryos and human cell-culture based experimental models. The first study showed that individually cultured preimplantation bovine embryos release EVs to their culture media, and their concentration and size profile are dependent on the quality and development stage of embryos. Subsequently, the functionality of preimplantation embryo-derived EVs were tested in the oviduct. It was observed that EVs from good quality embryos, but not the EVs from embryos of low developmental potential quality, could alter the gene expression of the oviduct. Among the up-regulated genes, many were interferon-stimulated genes of the interferon-τ pathway. Interferon-τ is considered the pregnancy recognition molecule in ruminant pregnancy. This finding suggests that the oviduct can serve as a biosensor of embryo quality. Finally, the functional specificity of embryonic EVs were investigated. It was observed that endometrium only react to embryonic EVs but not to the non-embryonic EVs. All these studies support the potential and specificity of embryo-derived EVs as a biomarker of embryo quality.https://www.ester.ee/record=b548409

    Influence of newly-synthesized chalcone derivatives on growth, biofilm production, and virulence factors expression of multiresistant Acinetobacter baumannii strains

    Get PDF
    Acinetobacter baumannii je nozokomijalni, multirezistentni patogen, koga karakterišesposobnost perzistencije na neživim površinama i mogućnost veoma brzog sticanja rezistencije naantibiotike. Danas su u svetu rasprostranjeni izrazito rezistentni sojevi protiv kojih u mnogimzdravstvenim ustanovama ne postoji efikasna terapija, a pronalazak alternativnih terapijskihpristupa je od izuzetne važnosti. Halkoni su jedinjenja sa potvrđenim antimikrobnim svojstvima ipokazanim različitim antivirulentnim aktivnostima. Ciljevi istraživanja ovog rada bili suodređivanje profila rezistencije, ispitivanje mogućnosti kontaminacije antiseptika i ispitivanjeprodukcije biofilma identifikovanih kliničkih izolata A. baumannii, kao i sinteza derivatahidroksihalkona i ispitivanje njihovih antimikrobnih i antivirulentnih aktivnosti protiv ovih izolata.Osetljivost izolata na antibiotike ispitana je kombinacijom difuzionih, dilucionih iautomatizovanih metoda, a identifikovani kolistin-rezistentni izolati dodatno su podvrgnutisekvenciranju celog genoma (WGS) i genetički su okarakterisani. Takođe, mehanizmi rezistencijena kolistin ispitani su primenom komparativne analize genoma i Real-Time kvantitativne lančanereakcije polimeraze (RT-qPCR). Time-kill test je primenjen za ispitivanje perzistencije uantisepticima, a nivo produkcije biofilma ispitan je pod različitim uslovima kultivacije in vitrostatičkom metodom uz bojenje safraninom. Derivati hidroksihalkona sintetisani su pomoću Claisen-Schmidt kondenzacije i njihove antimikrobne aktivnosti, samih i u kombinaciji sa antibioticima,ispitane su bujon-mikrodilucionom, Time-kill i Checkerboard analizom. Antivirulentne aktivnostiodabranih halkona procenjene su posredstvom uticaja na produkciju biofilma (monomikrobnog ipolimikrobnog), vijabilnost biofilmskih ćelija, ekspresiju motiliteta, gensku ekspresiju faktoravirulencije (OmpA, Bap i AbaI), adheziju A. baumannii na komponente ekstracelularnog matriksa(ECM), kao što su fibronektin i kolagen, i aktivnost sistema međućelijske komunikacije (Quorum-Sensing, QS).Klinički izolati A. baumannii gotovo uniformno bili su rezistentni na karbapeneme, a čakskoro 19% izolata bilo je rezistentno na kolistin, pripadajući tako ekstenzivno rezistentnom ilipanrezistentnom fenotipu. Izolati su pokazali sposobnost kontaminacije antiseptika i produkcijevelikih količina biofilma. Nutritivni sastav hranljivih medijuma značajno je uticao na nivoprodukcije biofilma, dok se visok nivo produkcije održao pri širokom opsegu različitih temperaturainkubacije i u prisustvu subinhibitornih koncentracija antibiotika. Sintetisani halkoni ispoljili suumerenu antimikrobnu aktivnost, pri čemu su metoksi-supstituisani derivati u proseku najjačeinhibirali rast. Takođe, zabeleženo je nekoliko sinergističkih interakcija halkona sa meropenemom,a inhibicija efluksnih pumpi predložena je kao potencijalni mehanizam. Halkoni su pokazalisposobnost značajne inhibicije motiliteta i produkcije biofilma, a metoksi-supstituisani derivat (o-OCH3) ispoljio je značajnu antivirulentnu aktivnost posredstvom nishodne regulacije ekspresijeompA, bap i abaI gena i inhibicije adhezije na komponente ECM. Na osnovu ovih rezultata, o-OCH3 halkon je identifikovan kao potentni antivirulentni agens protiv A. baumannii.Acinetobacter baumannii is a nosocomial, multiresistant pathogen, able to persist on abioticsurfaces and to rapidly acquire antibiotic resistance. Nowadays, highly resistant strains are widelydisseminated throughout the world, and the discovery of alternative therapeutic strategies is of utterimportance. Chalcones are compounds whose antimicrobial properties are well-known and forwhich different antivirulence activities have been demonstrated. The aims of this research were todetermine resistance profiles, to evaluate the possibility of antiseptic contamination, and to analyzethe biofilm production of identified A. baumannii clinical isolates, as well as to synthesizehydroxychalcone derivatives and to investigate their antimicrobial and antivirulence activitiesagainst these isolates.Antibiotic susceptibility of the isolates was tested by combination of diffusion, dilution, andautomated methods, and additionally, identified colistin-resistant isolates were subjected to wholegenome sequencing (WGS) and were genetically characterized. Also, colistin resistancemechanisms were explored by using comparative genome analysis and Real-Time quantitativepolymerase chain reaction (RT-qPCR). Time-kill test was used for the measurement of bacterialsurvival in antiseptics, whereas the level of biofilm production under different cultivationconditions was quantified by in vitro static method using safranin stain. Hydroxychalconederivatives were synthesized by Claisen-Schmidt condensation, and their antimicrobial activities,alone and in combination with antibiotics, were investigated using broth-microdilution, Time-kill,and Checkerboard analyses. Antivirulence activities of selected chalcones were evaluated based onthe impact on biofilm production (monomicrobial and polymicrobial), biofilm cell viability,motility, virulence factors (OmpA, Bap, and AbaI) gene expression, fibronectin- and collagen-mediated adhesion, and quorum-sensing (QS) activity.A. baumannii clinical isolates expressed extensive drug-resistant or pan-drug resistantphenotypes, being nearly uniformly resistant to carbapenems. Almost 19% of isolates were resistantto colistin as well. The isolates proved to be able to contaminate the antiseptic solutions and toproduce large quantities of biofilms. Nutritional composition of growth media significantly affectedthe level of biofilm production. In contrast, wide range of different incubation temperatures and thepresence of antibiotics at subinhibitory concentrations had little effect, and the bacteria managed tomaintain high level of biofilm production. Moderate antimicrobial activity was displayed bysynthesized chalcones, among which methoxy-substituted derivatives achieved greatest growthinhibition in average. Also, synergistic activity of chalcones and meropenem was present in severalcases, for which efflux pump inhibition was proposed as the potential mechanism. The chalconessignificantly inhibited motility and biofilm production, whereas methoxy-substituted derivative (o-OCH3) also displayed significant antivirulence activity, by downregulating the ompA, bap, and abaIgene expression and by inhibiting fibronectin- and collagen-mediated adhesion. It can be concludedthat o-OCH3 has been identified as a potent antivirulence agent against A. baumannii

    Investigating novel aspects of the blood-brain barrier using high resolution electron microscopy

    Get PDF
    Doctor ScientiaeThe blood-brain barrier (BBB) is a restrictive interface located between the blood circulation and the central nervous system (CNS), regulating the homeostatic environment of the neuronal milieu, by controlling the permeability of the cerebrovasculature. Currently, we cannot fully comprehend the regulatory features and the complexity of BBB morphology to allow for intervention clinically. The thesis consists of four publications. The methodology paper proposes a novel experimental design to visualize the morphological architecture of immortalized mouse brain endothelial cell lines (bEnd3/bEnd5). The brain endothelial cells (BECs) were grown on cellulose matrices and fixed in 2.5 % glutaraldehyde in preparation for visualization of the paracellular (PC) spaces between adjacent BECs, employing high-resolution electron microscopy (HREM), with vested interest in the morphological profile of the developing BEC. The second publication addresses and reports on the nanosized detail of BEC monolayer morphology utilizing high-resolution scanning electron microscopy (HR-SEM) and published the first descriptions of the extrusion of a basement membrane from developing in vitro BECs. Moreover, we categorized and discussed two types of nanotubule (NT) development specific for the establishment of the BEC monolayers. NTs can occur via nanovesicle extrusion onto the BEC membrane surfaces, which fuse, forming tunneling NTs (TUNTs) between adjacent BECs. Furthermore, cytoplasmic extensions of BEC membrane leading edges give rise to tethering NT (TENTs), which result in overlapping regions across the PC spaces, resulting in PC occlusion. BEC NT communication is illuminated in a third publication utilizing immunofluorescence microscopy, which reports on the molecular, cytoskeletal elements governing NT formation. This study shows, for the first time, f-actin and α-tubulin cytoskeletal proteins extending between the soma of the cells and NT cytoskeletal structures within an in vitro BBB model. Thereafter, the effects depolymerizing agents, Cytochalasin D and Nocodazole, were investigated on f-actin and α-tubulin cytoskeletal protein generation,functionality of NT morphology, cell division and permeability. For the first time, we show that f-actin possesses an additional function, key to tight junction, plaque protein organization. Moreover, it facilitates TENT formation, essential for cytoplasmic projection across PC spaces. Conversely, α-tubulin facilitates known functions: (i) transportation, (ii) cytokinesis, (iii) cellular division, and (iv) possesses a novel function as the molecular cytoskeletal backbone of TENTs, which facilitates BBB impermeability. A critical review evaluates past literature, in light of the current findings emanating from this study. The review critiques the concept of BEC cilia, which have been reported in the literature, comprised of tubulin and actin, but at low-resolution. In the light of our novel observations, nowhere in transmission electron microscopy do we observe cilia on the BECs, we postulate that NTs have been misnamed and mischaracterized as cilia. The thesis endeavors to elucidate the complexity of BEC nanostructures by examining the emerging role of the nanoscopic landscape of BBB development and the changing nature of BEC morphology, NT formation and associated cytoarchitectural underpinnings governing NT morphology. The research study attempts to, with a view to create new avenues for treating brain pathology, revolutionize our interpretation of barrier-genesis on a nanoscale

    Exploration of an alginate lyase targeting the biofilm in Pseudomonas aeruginosa

    Get PDF
    Pseudomonas aeruginosa is an opportunistic gram-negative bacterium capable of colonizing immunosuppressed patients such as those with cystic fibrosis. A P. aeruginosa infection could lead to mortality since some strains of the bacterium can change morphology by overproducing the exopolysaccharide alginate, becoming a mucoid variant that is more resistant to host defense and medical treatment. Many of the enzymes connected to the alginate machinery have already been characterized. However, the putative polysaccharide lyase (PL) family 7 alginate lyase PA1784 is poorly described in the literature, and new treatment methods for mucoid biofilm might be discovered by testing enzymes associated with alginate. Therefore, PA1784 (renamed PaAly7a) was first purified, and then activity assays were conducted to test the activity against substrates like sodium alginate, Psl and Pet in biofilm, and purified alginate from P. aeruginosa. In addition, the sequence to PaAly7a was analyzed by bioinformatics tools, and the predicted structure was compared to the PL7 alginate lyase, PA1167. The results from the activity assays showed no signs of activity. Nevertheless, a STRING analysis showed a connection between PaAly7a and a DNase, so DNA was tested as a substrate in a DNA-degradation assay. The assay showed signs of DNA degradation, but a DNase was identified in the sample after a proteomics analysis. In addition, the results revealed that PaAly7a formed inclusion bodies when overexpressed in E. coli and that the lack of activity could have been due to improper folding of the protein. However, the melting point analysis of PaAly7a-P2 showed a melting point, indicating that it might be correctly folded. Furthermore, the predicted structure of PaAly7a showed that the active site and the area around it were tighter and more negatively charged than the active site in the PL7 alginate lyase PA1167. Despite the lack of activity, these results contribute to further knowledge about PaAly7a and the goal of finding an alternative treatment for P. aeruginosa infections in patients with cystic fibrosis.Pseudomonas aeruginosa er en gramnegativ opportunistisk bakterie som er i stand til å kolonisere immunsupprimerte pasienter, slik som de med cystisk fibrose. En P. aeruginosainfeksjon kan føre til dødsfall siden denne bakterien kan endre morfologi ved å overprodusere eksopolysakkaridet alginat. Dermed blir en mucoid variant mer motstandsdyktig mot immunforsvaret og medisinsk behandling. Mange av enzymene knyttet til alginat i P. aeruginosa har allerede blitt karakterisert, men PA1784 (referert til som PaAly7a), som er antatt å være en alginat lyase i polysakkarid lyase familien 7, er dårlig beskrevet i litteraturen. Siden nye behandlingsmetoder for P. aeruginosa-infeksjoner muligens kan knyttes opp mot enzymer assosiert med alginat, har PaAly7a blitt renset og testet. PaAly7a ble testet ved bruk av forskjellige aktivitetsanalyser og ulik substrater slik som natriumalginat, Pel og Psl i biofilmer og renset alginat fra P. aeruginosa ble brukt. I tillegg ble sekvensen til PaAly7a analysert med forskjellige bioinformatiskeverktøy, og den antatte strukturen sammenlignet med en PL7 alginat lyase, PA1167. Resultatene fra aktivitetsanalysene viste ingen tegn til aktivitet. En STRING-analyse viste at PaAly7a hadde en forbindelse til DNase, DNA ble derfor testet som et substrat i en DNA-nedbrytningsanalyse. Analysen viste tegn på nedbrytning av DNA, men DNase ble identifisert i prøven ved en proteomikk analyse. I tillegg viste det seg at inclusion bodies ble dannet når E. coli overuttrykket PaAly7a. Feil foldet protein kunne derfor forklare hvorfor proteinet ikke viste aktivitet. Imidlertid indikerte smeltepunktanalysen til PaAly7a-P2 at proteinet var riktig foldet. Videre viste den predikerte strukturen til PaAly7a at det aktive setet og området rundt var trangere og mer negativt ladet, enn det aktive setet til PA1167. Til tross for manglende resultat av PaAly7a aktivitet, bidrar disse resultatene til en økt kunnskap om PaAly7a og målet om å finne en alternativ behandling for P. aeruginosainfeksjoner hos pasienter med cystisk fibrose.M-BIOTE
    corecore