Faculty of Pharmacy
Recent Submissions
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(2024)Mesencephalic Astrocyte-derived Neurotrophic Factor (MANF) is an evolutionary conserved protein vital for regulating various physiological processes, with potential therapeutic applications for many conditions. MANF is primarily localized within the ER lumen; however, its intracellular localization and cellular trafficking during pathological conditions remain unclear. It has been shown that MANF is upregulated by ER stress and plays a crucial role in mitigating the unfolded protein response (UPR) by interacting with ER transmembrane sensors and chaperone proteins; additionally, MANF has demonstrated attenuating effects on oxidative stress and improved mitochondrial function. Therefore, this thesis aims to uncover the cellular intricacies of MANF, examining its potential nuclear translocation and expression dynamics under ER and oxidative stress conditions. Here, we show that MANF can localize into the cell nucleus, and various stress conditions alter the expression dynamics and localization of MANF. We detected MANF and some other ER-resident proteins in the nuclear fractions of cells and rat liver tissue in steady-state conditions in vitro. We found that MANF may translocate into the nucleus under stress-induced conditions. Furthermore, we showed expression dynamics of MANF and some other ER-resident proteins upon ER and oxidative stress in vitro. Our results demonstrate the dynamic localization and expression of MANF in response to ER and oxidative stress, revealing its potential involvement in cellular responses under stress conditions. These findings not only pave the way for further research into the precise roles and mechanisms of MANF but also inspire new areas for investigation, offering potential therapeutic implications for conditions that urgently require novel innovative treatments for patients.
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(2024)The preservation of vaccines through an effective cold chain is critical to ensuring their potency and efficacy. This thesis investigates the utilization of a previously screened biopolymer developed in our laboratory for vaccine preservation under various temperature conditions. The study evaluates the stability of vaccines formulated with this biopolymer at 4°C, 22°C, and 37°C over a period of four weeks. The results, obtained from both in vitro and in vivo settings, demonstrate that the biopolymer effectively maintains vaccine integrity across these conditions, with the preserved vaccines retaining their protein expression even under stress tests. These findings indicate that the biopolymer not only supports the stability of vaccines within the conventional cold chain but also extends their viability outside traditional temperature constraints. This research underscores the potential of the biopolymer as a versatile solution for enhancing vaccine preservation, particularly in low-resource settings where maintaining strict cold chain protocols is challenging. Through a combination of experimental data and analysis, this thesis provides compelling evidence for the adoption of biopolymer-based formulations to improve global vaccine distribution and administration. Ultimately, the study contributes to the global health community's efforts to ensure that vaccines remain effective from production to administration, thereby improving immunization outcomes and public health.
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(2024)Since their discovery, β-lactam antibiotics have been an essential part of antibacterial chemotherapy. However, antibacterial resistance against β-lactam antibiotics via β-lactamases is a major challenge since it reduces the efficiency of current antibiotics. There are clinically available serine-β-lactamase (SBL) inhibitors, yet metallo-β-lactamase (MBL) mediated resistance is an arising research problem due to their ability to hydrolyze SBL-resistant carbapenems, and SBL inhibitors as well. N-sulfamoylpyrrole-2-carboxylates (NSPCs) are one of the MBL inhibitor chemotypes that displayed potent inhibitory activity against various subtypes of MBLs. This thesis focuses on the synthesis of novel NSPCs. Additional purification methods were applied in order to improve the current procedure. A large-scale synthesis of the starting material was conducted first. Thienyl, and halogen and methoxy substituted phenyl attached NSPC analogues were selected to be synthesized. However, the syntheses failed at the Suzuki coupling stage for the three compounds, and for the other two the purification of the final compounds was not successful. The thesis offers valuable insights into how various purification methods affect the yield at various stages of synthesis. Despite unexpected results, it also highlights the challenges in creating new NSPC analogues.
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(2024)Alzheimerin taudin riskiä lisää monet tekijät sekä mahdollisesti tietyt lääkkeet. Näiden riskien monitoroinnilla saattaa olla suotuisa vaikutus ennusteeseen. Näin ollen riskiä lisäävät lääkkeet olisi erityisen tärkeää tunnistaa ja lopettaa tai rajoittaa käyttöä korvaamalla muilla lääkeryhmillä tai lääkkeettömillä keinoilla. Pitkäaikaisesti käytössä olleiden lääkkeiden hallittu lopettaminen eli annosten pienentäminen tai lopettaminen vaatii varovaisuutta ja tarkkaa harkintaa, jotta vältytään lopetusoireilta eikä heikennetä elämänlaatua. Tämän Tutkielman tavoitteena oli tutkia, kuinka kognitiivisia haittoja aiheuttavia lääkeaineita voidaan lopettaa hallitusti, jotta vältytään irreversiibeleiltä kognitioon kohdistuvilta haitoilta. Tämä toteutettiin tutkimalla lääkevalmisteiden lakisääteisistä valmisteyhteenvedoista (SmPC), pakkausselosteista ja muista keskeisistä terveydenhuollon ammattilaisille suunnatuista tietolähteistä̈ löytyvää tietoa trisyklisten masennuslääkkeiden ja bentsodiatsepiinien hallitusta lopettamisesta. Tutkimusta varten tarkasteltiin kaikkia Suomessa huhtikuussa 2024 markkinoilla olleita lääkeaineita, joilla on merkittäviä antikolinergisia vaikutuksia. Lisäksi tarkasteltiin tuolloin markkinoilla olleita unilääkkeinä käytettyjä bentsodiatsepiineja. Näistä tutkimukseen sisällytetyt lääkeaineet valittiin sen perusteella, että niiden lopettamisessa tulisi noudattaa varovaisuutta (valittiin trisykliset masennuslääkkeet ja bentsodiatsepiinit). Tutkimus toteutettiin laadullisena sisällönanalyysinä, jossa aineistona käytettiin tutkimukseen valittujen lääkkeiden (amitriptyliini ja nortriptyliini, diatsepaami, oksatsepaami ja loratsepaami) valmisteyhteenvetoja (n = 5), pakkausselosteita (n=5), kansainvälisesti merkittävää hoitosuositusta: 1) American Psychological Association APA: Practice Guideline for the Treatment of Patients with Major Depressive Disorder, Yhdysvallat, 2) National Institute for Health and Care Excellence NICE: Depression in Adults: Treatment and Management, 3) NICE: Medicines Associated with Dependence or Withdrawal Symptoms: Safe Prescribing and Withdrawal Management for Adults, 4) The Maudsley: Deprescribing Guidelines Antidepressants, Benzodiazepines, Gabapentinoids and Z-drugs, Iso-Britannia sekä yhtä kansallista hoitosuositusta (Suomalainen Lääkäriseura Duodecim: Unettomuus Käypä̈ hoito -suositus). Tiedon määrä, sisältö ja laatu vaihtelivat tietolähteiden välillä. Trisyklisten masennuslääkkeiden hallitusta lopettamisesta löytyi selkeästi vähemmän tietoa kuin bentsodiatsepiinien hallitusta lopettamisesta. NICE:n hoitosuositukset täydensivät toisiaan lääkityksen hallitusta lopettamisesta. APA- ja käypä hoito -ohjeistukset sisälsivät tietoa yleisellä tasolla, mutta eivät sisältäneet yksityiskohtia lääkityksen hallitusta lopettamisesta. Muistin ja kognition kannalta huomattiin, kuinka trisyklisten masennuslääkkeiden valmisteyhteenvedoista ja pakkausselosteista puuttui kokonaan tietoa mahdollisista muistiin tai kognitioon kohdistuvista haitoista. Bentsodiatsepiinien mahdollisista muistiin tai kognitioon kohdistuvista haitoista taas löytyi jonkin verran tietoa, mutta tieto oli suhteellisen rajallista ja epätarkkaa.
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(2024)Parkinson’s disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the nigrostriatal pathway, resulting primarily in motor dysfunctions. The key neuropathological features in PD are intraneuronal inclusions, Lewy bodies (LB), resulting from aggregation and accumulation of misfolded α-synuclein (αSyn). Although it has not yet been undisputedly proven whether the accumulation of αSyn and formation of LBs is the initial cause for the loss of dopaminergic neurons or simply an epiphenomenon, several PD models have been successfully developed by inducing αSyn aggregation. One of the approaches utilizes pre-formed fibrils (PFFs) generated from recombinant αSyn monomers in vitro. The model is based on the discovery that exogenous PFFs induce αSyn aggregation in the affected cells, which eventually leads to formation of LB-like inclusions. In previous studies, neurotrophic factors (NTFs), such as brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), and mesencephalic astrocyte-derived neurotrophic factor (MANF) have been shown to reduce the formation of PFF-induced LB-like aggregates. Although the effects of exogenous NTFs have been studied in PFF-based models, the effects of PFFs on the endogenous levels of NTFs has not received much attention. Therefore, the aim of this study was to assess the effects of PFFs as well as cell maturation on the endogenous levels of BDNF, GDNF, and MANF in primary mouse embryonic midbrain dopaminergic cultures. The midbrain dopaminergic neurons isolated from E13.5 mice were cultured for 15 days, during which αSyn aggregation was induced with PFFs on day 8 in vitro (DIV8). Cell lysate samples were collected at three time points: on DIV0 1 hour after plating, DIV8 1 hour after addition of PFFs, and DIV15. The expression levels of BDNF, GDNF, and MANF mRNA were measured with quantitative PCR (qPCR). An unsuccessful attempt to optimize a TRI Reagent™-based RNA extraction protocol was also conducted during the project. The data suggested that the expression of BDNF and MANF is affected by cell maturation regardless of the absence or presence of PFFs. However, further research is needed to confirm the results. Additionally, GDNF was shown to be expressed at extremely low levels in mouse dopaminergic neurons. Lastly, the study revealed that the reference genes Gapdh and Hprt1 commonly used in qPCR are affected by cell maturation and PFFs, thus not suitable for studying PFF-treated mouse midbrain dopaminergic neurons.