Browsing by discipline "Pharmaceutical biology"
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(2019)Anisakiasis is a parasitic disease caused by larval nematodes of the genus Anisakis. Humans become infected by consuming contaminated raw or undercooked seafood products. Most human infections are caused by Anisakis simplex (A. simplex) complex. Currently there is no effective drug for this global emerging disease. Novel active compounds against the nematode are needed for drug development purposes. The research with A. simplex requires the isolation of the larvae from fish, which is time-consuming, unecological and uneconomical. Thus, the utilization of the model nematode Caenorhabditis elegans (C. elegans) in the research of A. simplex is considered in this study. Activities of Tea tree, Java citronella and Ho wood essential oils against C. elegans were studied. Aim of the assays was to examine whether C. elegans could be used as a model for A. simplex. Observed effects on C. elegans were compared to the previously reported effects on A. simplex. Activity of Tea tree and Java citronella essential oils against A. simplex was also examined to confirm previously reported activity. In addition, activity of six coumarins against A. simplex was investigated. The aim of the assays was to discover novel active compounds against the pathogenic nematode. Four coumarins were tested against C. elegans to examine possible comparable effects. Toxicity studies were performed in aquatic medium in a 6 well plate format (A. simplex) and in a 96 well plate format or in 1.5 mL Eppendorf tubes (C. elegans). Tea tree essential oil showed dose dependent activity against C. elegans, producing 100% mortality with the concentration 20 μL/mL after 24 hours exposure. Compared to A. simplex, two to three times higher doses were required to produce same degree of mortality in C. elegans. By contrast, Java citronella and Ho wood essential oils showed no significant activity against C. elegans. The activity of Tea tree and Java citronella essential oils against A. simplex was confirmed. The tested coumarins displayed no significant activity against the nematodes. Due to the contradictory results, further investigation about the suitability of C. elegans as a model for A. simplex is needed. Differences between the effective concentrations are probably caused by the differences in the biology of the nematodes, which result from the phylogenetic distance. Based on current results, the tested coumarins were excluded as potential antinematodal compounds against A. simplex, due to the lack of any significant activity on this model.
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(2020)Making the treatment of these infections even harder is the fact, that Chlamydia pneumoniae can produce persistent forms of itself, which are immune to antibiotic treatment. When the bacteria sense a stress factor, for example the presence of a β-lactam antibiotic or interferon γ, they start producing these persistent forms called aberrant bodies. When the stress factor is removed, the bacteria can switch back to their replicating form and start infecting the tissues again. It is also known, that C. pneumoniae bacteria will trigger persistence when the bacteria migrate from lung epithelia into monocytes. Interestingly the onset of this mode of persistence does not require any other triggers besides the invasion of the monocyte. These persistence mechanisms enable latent, quiet, and recurring infections. This master’s thesis aimed to study the coculture of lung epithelial (HL cells) and monocytes (THP-1 cells), and by utilising the magnetic separation method presented by Kortesoja et al, to find a positive control compound in the prevention of Chlamydia pneumoniae internalisation into the THP-1 cells for said protocol. In these cocultures the inhibitory effect of different compound groups such as lignans present in Schisandra chinensis plant, MAPK-inhibitors, and β2,2-amino acid derivatives in C. pneumoniae migration from HL cells to THP-1 cells was assessed. Statistic relevance was observed in JNK inhibitor SP600125, MAPKAP-kinase-2 inhibitor SB203580, and ERK1/2 inhibitor FR180204 compounds. These compounds inhibited the internalisation of Chlamydia pneumoniae into THP-1 cells in the cell coculture by 61,05 ± 16,63 % (p = 0,0001), 54,06 ± 16,02 % (p = 0,0002), and 36,76 ± 10,33 % (p = 0,009) respectively. SP600125 and SB 203580 compounds also had an inhibitory effect on the internalisation of C. pneumoniae into the THP-1 cells in a cell monoculture (39,98 ± 18,92 %, p = 0,026 and 37,89 ± 19,47 %, p = 0,035 respectively), whereas FR180204 had no statistical significance, even though it inhibited the internalisation of C. pneumoniae into the THP-1 cells in cell monoculture by 27,53 ± 21,17 %. From the compounds used in the experiments, only MAPK inhibitors had an effect in inhibiting the C. pneumoniae internalisation into the THP-1 cells. The most potent compound in said inhibition was the JNK inhibitor SP600125. JNK pathway has been thought to take part in chlamydial infections but only little research has been done. The results of this master’s thesis’ experiments support the thought of JNK enzyme taking part in chlamydial infections but determining how exactly it affects the infection cycle of C. pneumoniae bacteria still needs further investigation.
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(2018)Tämä tutkimus esittelee kasviperäisen nanokuituselluloosageelin (NFC; GrowDex®) arvioinnin kolmiulotteisena (3U) kasvualustana rintarauhasen organogeneesin mallinnuksessa. Tutkimuksen tavoitteena oli tarkastella kasviperäisen in vitro -kasvualustan aiheuttamaa solusäätelyä normaalissa rinnan epiteelisessä solulinjassa, sekä selvittää rintakudoksen rauhasrakenteiden muodostumisessa keskeisen laminiini 111:n (LAM-111) alustaan lisäyksen mahdollisia hyötyjä viljelmille. Tutkimuksen koeasetelmassa NFC:n edustamaa kasvunicheä arvioitiin ihmisen rintaepiteelistä eristetyllä -ja tyvikalvon proteiinikontaktien säätelystä riippuvaisella MCF 10A -solulinjalla. Solujen in vitro -nicheympäristön verrokkimallinnuksessa hyödynnettiin epiteelisen tyvikalvon proteiiniympäristöä edustavaa proteiinirikasta Matrigel™-2,5U -kasvualustaa. Viljelynäytteistä tehtiin aikapisteittäin valomikroskooppiset -sekä histologiset hematoksyliini – eosiini (HE) morfologian arvioinnit, e-kadheriinin, vimentiinin ja β4-integriinin ilmentymisten vasta-aine-analyysit, sekä β1-integriinin, Bim:in ja c-FLIP-L:n lähetti-RNA:n reaaliaikaiset PCR-analyysit. Analyyseissä keskityttiin tarkastelemaan rintarauhasen epiteelin polarisoitumistapahtumassa havaittavaa solusäätelyä ja proteiinien eritystä. LAM-111 -lisän havaittiin edistävän jossain määrin NFC:ssä viljeltyjen sferoidien sisämorfologian kavitaatiota sekä eritettyjen proteiinien sijoittumista sferoidien pintarakenteisiin Matrigel™ -kontrollinäytteiden kaltaisesti, muttei yksinään riittänyt tuottamaan Matrigel™ :ssä havaittua viljelmien homogeenisyyttä. Kokeen natiivi-NFC:ssä sekä NFC-LAM-111:ssä kasvaneiden sferoidien PCR-analyyseissä havaittiin polarisaatiotapahtumaan liittyvää solusäätelyä viljelmien loppuvaiheessa päivänä 28, poiketen vastaavan PCR profiilin ilmentymisestä Matrigel™ -viljelmissä jo päivänä kolme. NFC -olosuhteissa havaittiin myös Matrigel™ -viljelmistä puuttuvia ylimääräisiä, epiteelisiltä vaikuttavia rakenteita, joiden määritteleminen vaatii lisätutkimuksia. NFC todettiin jäykkyyden suhteen helposti muokattavaksi sekä mahdollisesti kudoksen mekaanisia ominaisuuksia jäljitteleväksi 3U -kasvualustaksi. Tämän kokeen tuloksien perusteella muokkaamatonta NFC:tä voidaan ehdottaa soveltuvaksi kasvualustaksi tyvikalvoproteiinien säätelystä riippumattomille solutyypeille, sekä solutyypeille, jotka kykenevät tuottamaan ympärilleen oman kudostyypillisen proteiiniympäristönsä. Kliiniseen käyttöön kelpuuttavat standardivaatimukset täyttävä NFC vaikuttaa lupaavalta materiaalilta räätälöitävien in vitro -kasvualustojen suunnitteluun, ja mahdollisesti tarjoaa rakenneosiltaan tarkasti määritellyn, xenovapaan, ja proteiinilisillä eri solutyypeille säädettävän in vitro -kasvunichen tulevaisuuden jatkotutkimuksiin.
Now showing items 1-3 of 3