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  • Nevala, Laura (2010)
    The objective of the research was to study the effects of different polymers, sugars, and cell handlings on the viabilities of ARPE-19 and ARPE-19-SEAP-2-neo cells after freeze-drying. Also the residual moisture content of the freeze-dried samples and the amount of intracellular sugar after incubation in trehalose medium were measured. The mixtures used in the study contained different polymers (e.g. polyvinylpyrrolidone, alginate, polyethylene glycol) and sugars (sucrose, trehalose and mannitol). Some cells were incubated or heated in trehalose medium before freeze-drying in order to increase the amount of intracellular sugar. The aim of the heating was also to increase the heat shock protein formation in the cells. The samples were mainly freeze-dried in 24 well plates. The same freeze-drying parameters were used in all freeze-drying runs. The freeze-drying cycle lasted 38.5 hours (freezing 2.5 h, primary drying 32 h, and secondary drying 4 h). In the freezing stage the samples were froze to -40 °C and the freezing rate was 1 °C/minute. In the primary drying stage the shelf temperature was mainly -35 °C and the pressure was 150 mTorr. The viabilities of the samples after freeze-drying were determined by measuring the fluorescence after 3 and 6 hours from addition of the Alamar Blue indicator dye. The residual moisture contents were measured by thermogravimeter. According to the results, mixture containing glycerol (9%) and PEG 10 000 (18%) was the best lyoprotectant in the study (70% viability after 3 hours). However, the viability decreased significantly (24% viability) in the measurement after 6 hours. Similar viability decrease was observed among all lyoprotectant mixtures used in the study. Extracellular sugars rarely had positive effect on the viability results. The 12 days incubation in 150 mM trehalose medium before freeze-drying affected positively to the post freeze-drying viability. Shorter incubation time or heating did not induce the same effect. Intracellular sugar measurements revealed, that the amount of intracellular trehalose was multiple after 12 days of incubation in 150 mM trehalose medium compared to the cells that were not incubated. The residual moisture contents of the samples varied between 0-7%. The residual moisture content of the sample containing glycerol 9% and PEG 10 000 18% was 1,5%.
  • Sihvola, Otto (2024)
    Finland has long traditions on the use and development of digital healthcare systems. This infrastructure, covering the entire population, has enabled the utilization of electronic healthcare records (EHR) collected in various registers for secondary purposes, such as research, development, and innovation (RDI) activities. Following the emergence of this activity, an operational environment supporting the reuse of data developed between these registers and data users, enabling diverse research activities. The Secondary Use Act, prepared on the basis of the General Data Protection Regulation (GDPR) and which came into force in May 2019, triggered many reforms for this operating environment. According to experts from various stakeholder groups, the reforms were seen as both beneficial and challenging, as observed in a small number of related studies already conducted. The purpose of this study was to investigate the current state of the impacts brought by the Secondary Use Act by mapping the views and opinions of representatives from different expert groups on key changes in the operating environment from their perspective and identifying potential areas for development. The study was conducted in early 2024 as a semi-structured interview study. The interviewees consisted of experts (n=8) from various stakeholder groups familiar with the operating environment for the secondary use of real-world data. The interviewees had extensive and significant experience with either the Secondary Use Act or its operating environment. Six distinct themes emerged from the material: data management in national registers, data utilization in research, the national process and guidance, the preparation process and culture of the Secondary Use Act, internationality, and actors and their equality. To identify new and significant observations, a comparison with previous research findings was conducted, ultimately identifying key factors that still challenge stakeholders. These factors related to inequalities among stakeholders, restrictions due to data protection requirements, lack of guidelines, and data harmonization. The results confirm the previous studies’ observations and highlight the particularly challenging areas for stakeholder groups in the current time. Overall, opinions about the data permit authority were mostly positive, while there was relatively more distrust towards register holders and legislators. Based on the study, there are clear grounds for developing the Secondary Use Act, but its timing is influenced by the progress of international projects, such as the EHDS model. Future research should increasingly focus on joint European projects to integrate Finland into the international operating environment.
  • Hautaniemi, Mikaela (2012)
    In pharmaceuticals amorphous state can be obtained either intentionally or unintentionally. Intentional production is used, for example, to improve the dissolution of poorly soluble compounds, to stabilize the structure of proteins, or to improve the mechanical properties of excipients (e.g., lactose). Unintentional introduction of amorphous phases can result from general manufacturing procedures of pharmaceuticals, such as coating, granulation, drying, milling, and compression. The presence of amorphous regions, even in small quantities, can exhibit a significant influence on the physical and chemical stability of pharmaceutical products. Molecular mobility in formulation with amorphous content is believed to be the key factor of their stability. Therefore, evaluating of molecular mobility is an important step in pharmaceutical product development. The aim of this study was to estimate molecular motions in amorphous disaccharides using calorimetric approach at temperatures below the glass transition temperature (Tg), where relaxation process is very slow as compared to the time of experiment. When temperature is low enough, the initial relaxation time parameter (τi) can be used as an estimate for relaxation process on the timescale of pharmaceutical product shelf life. The results of the present study revealed similar trend in stability of amorphous forms for the disaccharides (sucrose experiencing the fastest structural relaxation), which can be assumed on the basis of Tg alone, where higher Tg would result in more stable glassy state (Tg of sucrose is the lowest). Storage temperature of Tg - 55oC or lower would suffice for amorphous trehalose, melibiose and cellobiose to achieve at least 2 year's relaxation time, while for sucrose the temperature is Tg - 70oC. Fragility has been used as a helpful mean for classifying amorphous materials. All the compounds can be classified as fragile. Fragility ranking in the present study contains some degree of uncertainty, while 3 different approaches revealed somewhat different results for ranking the disaccharides. The variation in the results can be attributed to the overall sensitivity of DSC. The method described in the present study is quite difficult to apply without supportive information from other techniques. The results, obtained with the method, are very dependent on the slope in plotting ln q vs. 1/Tg, and even small fluctuations in the estimation can lead to different fragility values and consequently to different relaxation times. However, the final results reveal values for relaxation times well below Tg, which are in reasonable agreement with modern theoretical understanding of glassy state dynamics.
  • Korventausta, Susanna (2022)
    Etätyö yleistyi maaliskuussa 2020 äkillisesti COVID-19 pandemian seurauksena maailman terveysjärjestö WHO:n suosituksesta. Etätyö on ollut ennen koronapandemiaa harvinaista lääketeollisuudessa, joten etätyötä lääketeollisuudessa on tutkittu hyvin vähän. Etätyön on arvioitu jäävän pysyväksi ratkaisuksi, joten on ajankohtaista tutkia etätyön soveltuvuutta ja tehokkuutta lääketeollisuudessa. Etätyöntekijöiden tuottavuus kasvaa yleensä huomattavasti. Työpaikalla koetaan jatkuvasti keskeytyksiä, melua ja muita häiriötekijöitä, joiden lisäksi työmatkat kuormittavat työntekijöitä. Etätyöntekijät säästyvät suurimmalta osalta näistä ongelmista, jolloin suurempi osa heidän työpäivästään kuluu varsinaiseen työntekoon. Valtaosa etätyöntekijöistä tekee etätyötä osan työajastaan. Tutkimuksen tavoitteena oli selvittää kokemuksia etätyöstä, etätyön soveltuvuutta ja etätyön tehokkuuteen vaikuttavia tekijöitä lääketeollisuudessa. Tutkimus on toteutettu Orion Oyj:n Suomen toimipisteissä. Tutkimuksen toteuttamistapa oli kvantitatiivisen ja kvalitatiivisen kyselytutkimuksen yhdistelmä. Yhdistämällä kvantitatiivisia ja kvalitatiivisia kysymyksiä pyrittiin saamaan tarkempia tietoja kuin pelkällä kvantitatiivisella tutkimuksella voitaisiin saada. Kysely oli avoinna 15.11.–26.11.2021. Vastausprosentiksi saatiin 34,9 %. Etätyön merkittävimmiksi hyödyiksi havaittiin työ- ja vapaa-ajan joustavampi yhteensovittaminen ja se, että etätyössä keskittyminen on parempaa. Kommunikaation koetaan onnistuvan hyvin etätyössä, mutta kasvokkain tapahtuvaa kommunikaatiota pidetään myös tärkeänä. Esimerkiksi kehitys- ja ideointipalaverit olisi hyvä järjestää mahdollisuuksien mukaan kasvokkain. Lisäksi hiljaisen tiedon siirtyminen on vähäisempää etätyössä. Etätyö soveltuu hyvin tutkimus- ja tuotekehitystyöhön, eikä sen koeta heikentävän merkittävästi kykyä innovoida. Tulosten perusteella etätyötä haluttaisiin tehdä enemmän kuin 40 % työajasta ja etätyötä pidetään tehokkaana työskentelytapana. Kyselyssä ei selvitetty, minkälaisia etätyömääriä vastaajat ovat tehneet vahvan etätyösuosituksen aikana. Osa vastaajista on saattanut olla muita enemmän lähitöissä, mikä voi vaikuttaa tuloksiin. Saadut tulokset olivat kuitenkin samansuuntaisia kuin aikaisemmissa tutkimuksissa. Suurin osa tähän kyselyyn osallistuneista oli erittäin kokeneita ja työnsä hyvin osaavia työntekijöitä, mikä voi lisätä etätyömyönteisyyttä tuloksissa.
  • Mäkinen, Heljä (2022)
    Municipal case management is an activity that assesses various functional capacity indicators, utilizing the elderly’s state of health and coping in everyday life. The goal of case management is to refer clients to suitable services, such as home care or a doctor's visit. The problems related to drug treatments are only superficially reviewed. The involvement of a pharmacist in the assessment of case management would provide an opportunity to address the problems of pharmacotherapy and to provide adequate support for the implementation of pharmacotherapy. In this thesis, a remote service of a pharmacist was piloted for new clients over the age of 65 living at home as part of case management. Pharmacist reviewed medications remotely using medication risk management checklist LOTTA. The study examined the suitability of the LOTTA for medication reviews and the problems associated with medications of the elderly participating in case management. In addition, the suitability of pharmacovigilance assessments as a remote service as part of a comprehensive assessment of functional capacity and coping with everyday life was examined. The research material was collected at the case management unit of the city of Turku. The study involved 50 volunteer Finnish-speaking customers over the age of 65, for whom were assessed for a case management at Turku's case management unit. In addition to the assessment of normal case management, two pharmacists with comprehensive medication review qualifications reviewed medications using the medication risk management checklist LOTTA. Subjects were interviewed by telephone. If the pharmacist estimates that the subject will benefit from a multi-professional comprehensive medication review, the physician and pharmacist collaborated to conduct a review using a videophone application. Subjects background information, responses, observations made by pharmacists, and actions taken by physicians were recorded on an electronic form and analyzed. The mean age of the study participants (n = 50) was 82 years (range 67–98). Of these, 36 were women (72%) and 14 were men (28%). Most subjects were multidrug-treated (average medication 10.3, range 3–28). Each subject had at least one drug can be used with consideration for use in the elderly, as defined in the Fimea Drug 75 + database (Class C). 30% of subjects did not have a medication list and 34% reported lack of regular medication monitoring. 96% of the subjects had experienced a symptom on the LOTTA list that repeatedly interferes with their lives. The most common of these were problems such as constipation (54%). Pharmacists proposed changes for medication for 96 % of subjects. The most common proposed change was a change in the time of dosing (46%). Pharmacists estimated that 14 (28%) subjects would benefit from a multi-professional comprehensive medication review. In these cases, pharmacists made an average of 8.1 proposed changes for the physician, and the physician made an average of 6.9 changes for each subject. The most common challenges in coping with medication were symptom, which may be due to adverse drug reactions, a lack of follow-up to medication, and the absence of a treating physician. The results suggest that medication should be reviewed during the case management. The LOTTA list made it possible to identify and address the pharmacological problems of the elderly. The participation of a pharmacist in the assessment of the need for a multi-professional service remotely was possible, but it must be further developed. More research is needed on the benefits of multi-professional case management with a larger sample size.
  • Hautala, Jonna (2018)
    Appetite regulation is a complex process involving regulation of energy homeostasis and the rewarding nature of food. Abnormalities in appetite regulation lead to obesity and eating disorders which are challenging to treat with medicines. Especially obesity is an increasing public health problem and drug development against it is a current subject in research. Hypothalamus is the most important brain area related to appetite regulation. Also, the basal forebrain and the amygdala which are part of the reward system in the brain, contribute to the appetite regulation. There is cholinergic innervation from the basal forebrain to the amygdala and most of the cholinergic activity in the amygdala is originating from the basal forebrain. It is known that the cholinergic system inhibits appetite but there is still no research about impact of cholinergic projections between these two brain areas. Aim of this study was to find out if the cholinergic projections from the basal forebrain to the amygdala effect on appetite regulation. The study included two stereotactic surgery. In the first surgery the mice (n=14) received injections to the basal forebrain that contained genetic materials for DREADDs in AAV. DREADDs appeared in the cholinergic cells of the basal forebrain and emanated within their axons to the amygdala. In the second surgery cannulas were placed to the amygdala. CNO was injected through the cannulas to the amygdala to cause the DREADDs activate or inhibit the cholinergic cells. As a control, mice received vehicle. The feeding experiments were performed in normal conditions or after food restriction and there were either food or sugar pellets available. The pellet dispenser monitored how many pellets mice ate during the six hours after the CNO or vehicle treatment. The success of virus injections was checked after the feeding experiments by antibody dyeing. In any conditions there was no significant differences in the results due to DREADDs and treatment. Because of the small group sizes and dispersion of the results no final conclusions can be made but the additional research about this topic is needed.
  • Tenhola, Heli (2013)
    Prostate cancer is the most common cancer among men in Finland. Today, new prostate cancers are diagnosed in an early phase of the disease when the cancer is still local and effectiveness of the treatments good. There are many effective treatment options for localised prostate cancer but all of them cause multiple side effects. No comprehensive information about prostate cancer patients' experiences, treatment consequences and outcomes has been available. National Institute for Health and Welfare (THL) carried out a nationwide survey to prostate cancer patients diagnosed in 2004. The questionnaire was sent by mail in 2009. Patients were asked to report their experiences during the diagnosis and treatment selection (prostatectomy, hormone therapy, external beam radiation, brachytherapy, surveillance). They were asked about amount and harmfulness of side effects and satisfaction with outcome of the treatment. An association between the side effects and satisfaction with the treatment outcome was also studied. A total of 1239 responses were accepted for the study (response rate 73%). All treatments caused several side effects, and up to half of the patients had some adverse effect still present at the time of the survey. Most of the side effects concerned urinary or sexual dysfunction, in external beam radiation also bowel dysfunction. All treatments caused sexual dysfunction, but radiation therapy less than the other treatments. Side effects caused by hormone therapy were specific for this treatment, like hot flashes and mood disturbances. Patients treated with prostatectomy were least satisfied with the outcome of the treatment and dissatisfaction was mostly associated with sexual and urinary dysfunction. Patients treated with radiation therapy were most satisfied with the outcome. Urinary and bowel dysfunction impaired the satisfaction in patients treated with external beam radiation. All prostate cancer treatments cause plenty of potentially harmful side effects that may be challenging for psychological and psychosocial well-being of the patients. Thus, both treatment modalities and means to support well-being of the patients should be developed further. Actions that enhance well-being and prevent and relieve side effects should be an essential part of standard clinical procedures for every prostate cancer patient.
  • Nieminen, Jonna (2010)
    Prostate cancer is one of the most common cancers in the developed countries. Prostate cancer is slowly progressing cancer but can transform into aggressive disease and metastasize. Metastases are the major cause of mortality. Androgens play an important role in the pathogenesis of prostate cancer and prostate tumors are usually dependent on androgens. Thus the aim of the treatment is to eliminate testicular androgens by surgical or medical castration and/or block the effect of androgens on the prostate with antiandrogens. Prostate cancer and new therapies to treat the disease are being investigated vigorously. Numerous in vivo models of prostate cancer have been developed. Androgen responsive animal models mimic prostate cancer more closely. There are many animal species that may be used to model prostate cancer but mouse is no doubt the most useful. Tumor models can be created by inoculating human cancer cells or solid parts of tumors into immune deficient mice. Orthotopic prostate tumor models reflect the abnormal cancer cell-stroma interactions occuring in prostate cancer. Transgenic mouse models are becoming more and more common in the research of prostate cancer. Transgenic models are able to model the initiation and progression of the disease more realistically. Growth of the orthotopic tumor is difficult to monitor without measuring serum prostate specific antigen (PSA) concentrations or using specific imaging methods. Imaging techniques, such as optical imaging, are being utilized in different in vivo models of prostate cancer. The objective of the experimental part of this thesis was to optimize bioluminescence imaging method in androgen responsive cell line LNCaP-luc2 in orthotopic model of prostate cancer. Bioluminescence imaging is based on a reaction catalyzed by a luciferase which is expressed by the tumor cells. In the ATP-dependent reaction luciferase enzyme oxidizes its substrate, luciferin, and produces light. In addition, the purpose of this study was to examine the effects of medical therapy and castration on tumor growth. Bioluminescence imaging enabled noninvasive, real-time and longitudinal monitoring of the growth of prostate tumors in this model. Quantification of the tumors with bioluminescence measurement was faster than with ultrasound sonography. It was also possible to monitor the growth of the tumors more often with bioluminescence imaging than with PSA measurements. Bioluminescence imaging was found to correlate better with serum PSA concentrations than with the actual size of the tumor. However prostate tumor size was noted to correlate better with PSA concentrations than with bioluminescence imaging in this study. Medical treatment or castration was found to have no effect on the size of the tumors when measured with bioluminescence imaging. The larger size of the tumors than expected was the probable reason for this. Bioluminescence imaging is not suitable for large or necrotic tumors because this imaging method can only be applied in living cells. In addition, a successful luciferin injection is essential for the proper utilization of bioluminescence imaging in this model. More studies are needed to validate the model for example in proving the effects of the medical therapies.
  • Nguyen, Thuy (2023)
    Microcrystalline cellulose (MCC) is a purified, partially depolymerized cellulose, which is obtained by treating α-cellulose with mineral acids. Ever since the first microcrystalline cellulose was commercialized, different grades of microcrystalline cellulose have widely been used in the manufacture of solid dosage forms, such as tablets. MCC obtained from different sources will exhibit different physico-chemical properties, including moisture content, degree of polymerization, crystallinity, and particle morphology. In wet granulation, microcrystalline cellulose can be used as a filler, binder, and disintegrant. Recently, Aalto University has introduced a novel microcrystalline cellulose obtained from renewable raw materials by an integrated process, which has a short retention time, low energy and chemical consumption. However, very few studies have evaluated the use of AaltoCellTM as an excipient in solid dosage forms. The objective of this study was to evaluate the filler properties of three grades of AaltoCellTM to prepare paracetamol tablets with 50% (w/w) drug load and compare AaltoCellTM with a commercial microcrystalline cellulose, Vivapur 101. Due to the poor flowability of paracetamol and the experimental microcrystalline celluloses, it is challenging to direct compress tablets from paracetamol and microcrystalline mixtures. Thus, the powder mixtures were granulated by high-shear wet granulation method to improve the flowability. After the granulation, the formulations were characterized for particle size distribution, morphology and powder flow. Carr’s index Hausner ratio and angle of repose were calculated to evaluate the flowability of the formulations. In addition, an image-based analysis of powder flow was performed. A rotary tablet press equipped with single punches of 9 mm diameter was used to compress tablets. To evaluate the quality of tablets, European Pharmacopoeia tests of friability, disintegration, uniformity of mass, uniformity of content and dissolution were conducted. The AaltoCellTM A and Vivapur 101 formulations had the smallest particle size, whereas the AaltoCellTM B had the largest particle size. According to Carr’s index and Hausner ratio, the flowability of AaltoCellTM powders and Vivapur 101 varied from poor to very, very poor. After the granulation, the flowability of AaltoCellTM B and AaltoCellTM C were classified as good, while AaltoCellTM A and Vivapur 101 formulations had fair flowability. However, the results were conflicting with the flowability index values obtained in the image-based analysis. According to the results, the AaltoCellTM tablets complied with all criteria of European Pharmacopoeia and were comparable with Vivapur 101 tablets. The average tablet weight deviated ± 3.2% from the target weight. The variations in weight and drug content were small, as indicated by low RSD values. The disintegration time of the AaltoCellTM tablets was between 1-8.5 minutes. In addition, the AaltoCellTM tablets had fast dissolution with 78-84% of paracetamol released within 1 minute. Overall, AaltoCellTM is a promising excipient for use as a filler in tablets. In further studies, characterizing the powder properties, such as morphology, surface properties and hygroscopicity, would provide a better understanding of the properties of AaltoCellTM.
  • Mörtengren, Ariel (2016)
    The field of stem cell research is hotter than ever, because still today, the goal for easily achievable stem cells for the use of tissue engineering and stem cell therapies, is yet to be achieved. Also, human stem cell based test systems are potential replacements of present animal test models. The ongoing obesity epidemic creates pressure for scientists to resolve the causes behind it. One way of approaching the problem, is the study of adipogenesis with the use of a in-vitro cell model. This have already been done for a while, with rodent based cell models, but the present study took the human obesity research a bit closer to its subject by using humane adipose tissue derived mesenchymal stem cells (hASC). Also, the adipogenic induction is executed with a human adipose tissue extract (ATE). Epidemiologically, the rise in obesity rates correlates at some level, with the occurrence of known endocrine disrupting chemicals in our environment. These include e.g. some pesticides and plasticizers, such as tributyltin (TBT), bis(2-ethylhexyl)phthalate (DEHP) and bisphenol A (BPA). In the present study, the effects of a variety of concentrations, ranging from 50nM to 100µM of BPA, on ATE induced adipogenesis of hASCs, was studied. The accumulation of triglycerides - a key parameter for adipogenesis - is evaluated with the use of oil-red-o (ORO) staining and photometric measurements. A set of tests was executed to find out if BPA possesses adipogenic, synergistic or antiadipogenic properties in this particular test system. No significant antiadipogenic, nor synergistic effects were seen. Some antiadipogenic effects were seen throughout the study, but without any dose-dependence. This study also showed need for further development of the test. ORO staining needs to be further standardized to increase accuracy, different batches of ATE may cause variation in the results. All and all the test system is relatively easily modified and when fully functional, it is a great tool for screening for substances affecting our adipose tissue, and also for enhancing our knowledge on human adipogenesis in whole.
  • Dang, Thu Ha (2023)
    Immune checkpoint inhibitor (ICI) therapy aims to enhance the endogenous immune response against tumour cells, and it has become a potent treatment option for various types of cancers. Despite the promise of ICIs, most patients do not respond to the treatment. The primary limitation of ICI therapy is the immunosuppressive tumour microenvironment (TME), which is characterised by the lack of tumour- infiltrating cytotoxic T cells (CTLs) and the presence of immunosuppressive cells, such as tumour- associated macrophages (TAMs). A promising immunotherapeutic strategy that can promote antitumor immunity is oncolytic virus (OV) therapy. OVs can selectively replicate in and kill cancer cells, leading to the release of immunostimulatory molecules. These molecules can induce local inflammation and prime and recruit CTLs to the tumour site. In addition, OVs can also be used as a delivery platform for immunostimulatory transgenes that can further enhance the activation of anti-tumour immune response and help to overcome the immunosuppressive TME. Another strategy used to support anti-tumour immune responses and overcome immunosuppressive TME is epigenetic therapy. Epigenetic therapy can reprogram both cancer and immune cells towards a less immunosuppressive phenotype, thus helping to overcome the limitation of immune checkpoint therapy. The aim of this study was to generate a novel oncolytic adenovirus armed with epigenetic modifying transgene (EpiCRAd) to overcome the immunosuppressive TME and enhance the anti-tumour immune response. We tested its efficacy and immunogenicity in vitro and in vivo using a murine triple-negative breast cancer model. We demonstrated that EpiCRAd was able to modulate the epigenome of cancer cells without affecting viruses’ infectivity. Upon examining the potential effect of EpiCRAd on cancer cells, we observed that epigenetic regulation did not notably influence the expression of MHC class I and PD- L1 proteins, both of which play a role in the immune evasion mechanism of tumour cells. In addition, the in vivo experiments show that EpiCRAd controls tumour growth the best, especially together with an immune checkpoint inhibitor, suggesting that the virus was able to create an immune microenvironment more favourable for anti-tumour response. Interestingly, the TAM infiltration in the TME seems to reduce after treatment with EpiCRAd. Overall, the combination of epigenetic therapy with oncolytic virotherapy has shown promising results in converting immunotherapy-resistant tumours into immunotherapy-responsive tumours. Our findings provide valuable insights into the effect of EpiCRAd on cancer and immune cells. This study encourages exploring the use of epigenetic cancer remodelling and oncolytic viruses for cancer immunotherapy.
  • Jalonen, Milla (2020)
    There are significant inter-individual differences in the effects of drugs. These differences can be caused by, for example, other diseases, adherence to treatment, or drug-drug interactions. A drug-drug interaction can lead to an increase in the concentration of the active substance in the circulation (pharmacokinetic interactions) or a change in the effect of the drug without changes in plasma concentration (pharmacodynamic interactions). A drug-drug interaction can change the efficacy of a drug or affect the adverse drug reaction profile. The individual’s genetic background, such as diversity in drug-modifying enzymes (polymorphism), also has an effect on the efficacy and the risk for adverse drug reactions of some drugs. A pharmacogenetic test can be used to study how genetic factors affect drug treatments. The aim of this master's thesis was to examine the possibilities of personalized migraine pharmacotherapy from the perspective of pharmacogenomics and drug-drug interactions. Four online drug-drug interaction databases available in Finland were compared. Inxbase is the most widely used interaction database by physicians in Finland and it is also integrated into Finnish pharmacy systems. Other databases used in this study were the international professional database Micromedex as well as Medscape Drug Interaction Checker and Drugs.com Drug Interactions Checker. The latter two are open-access databases available for healthcare professionals and patients. Interaction searches were conducted in the selected databases between acute and prophylactic drugs used for the treatment of migraine (e.g. bisoprolol-sumatriptan). Fourteen acute and 12 prophylactic drugs were selected for this study based on the Current Care Guidelines in Finland (Käypä hoito), and the data were collected in Excel spreadsheets. The first search was completed in December 2019 and the second search in March 2020. In this study, many potential interactions were found between acute and prophylactic drugs used to treat migraine in Finland. For more than half of the drug pairs studied, a potential interaction was found in at least one of the databases. There were significant differences between the interaction databases regarding which interactions the database contains and how the severity of the interactions was classified. Of the interactions found, only 45% were found in all four databases, and each database contained interactions that were not found in the other databases. Even very serious interactions or drug pairs classified as contraindicated were not found to be consistently presented across all four databases. When selecting drug treatment for a migraine patient, potential drug-drug interactions between acute and prophylactic drugs as well as the patient's genetic background should be considered. Individualizing migraine treatment to achieve the best efficacy and to reduce the risk for adverse drug reactions is important because migraine as a disease causes a heavy burden on individuals, healthcare, and society. Pharmacogenetic tests particularly developed to help choosing migraine treatment are not yet available, but tests are available for few other indications in both public and private healthcare. The use of these tests in clinical practice will increase as physicians’ pharmacogenetic knowledge and scientific evidence on pharmacogenetic tests increase. Utilization of pharmacogenetic data requires that test results are stored in electronic health records so that they are available in the future, when changes are made to drug treatment of individuals. More studies are warranted to better understand the clinical impact of pharmacogenomics and drug-drug interactions in migraine care.
  • Paakkunainen, Jonna (2023)
    Parkinson’s disease is a progressive neurodegenerative disorder which is commonly treated with Levodopa (L-dopa) and Dopa Decarboxylase (DDC)/ Catechol-O-methyltransferase (COMT) inhibitors. The main problem with this treatment is the intestinal conversion of L-dopa to dopamine despite DDC and COMT inhibition which probably occurs by the Tyrosine Decarboxylase (TyrDC) of intestinal bacteria. This study aims to find new inhibitor molecules that would have dual inhibitory effects towards both DDC and TyrDC enzymes. Currently, available DDC inhibitors cannot inhibit the bacterial TyrDC enzyme. A recently found TyrDC inhibitor (S)-α-Fluoromethyltyrosine (AFMT) is not able to inhibit the human DDC enzyme, respectively. The dual inhibition of both decarboxylases could reduce the dosing frequency and side effects related to L-dopa. In addition, the object of this study is to produce the human DDC enzyme by DNA recombinant technique as well as develop and optimize a biochemical DDC inhibition assay to study the effect of selected small molecule compounds towards inhibition of DDC and L-dopa conversion in E. faecalis model by previously developed cell-based assay. The human DDC was successfully produced in a TB medium with a yield of 1.8 mg/mL. The Km value of DDC for L-dopa was found to be 34 μM which indicates a high affinity for L-dopa. In the optimization of the DDC inhibition assay, the sample volume of 80 μL and incubation time of 3 h with detection reagent was found to give the highest fluorometric signal with sufficient robustness. In the initial screening of test compounds, 14 % of the compounds (n=59) were classified as active towards human DDC, while 31 % of the compounds were active towards L-dopa conversion in the E. faecalis model. Of those compounds, five were having dose-dependent dual inhibitory effects, but the IC50 values of them were higher compared to either carbidopa or AFMT. The most effective compounds were 8009-2501 (IC50 37 μM in E. faecalis model and 19 % inhibition at 1000 μM towards DDC enzyme) and 8012-3386 (IC50 248 μM in E. faecalis model and 37 % inhibition at 1000 μM towards DDC enzyme). However, this study confirms the possibility to find dual decarboxylase inhibitors. By optimizing the structures as well as investigating the mechanism of action, selectivity, and structure-activity relationships of the most active compounds, it is possible to find more effective dual inhibitors in the future.
  • Vainionpää, Teea (2016)
    Oncolytic viruses have been extensively studied for the treatment of cancer. They are genetically engineered viruses, which are able to selectively infect and kill the cancer cells causing no harm to normal cells. Adenoviruses are the most commonly used viruses in the gene therapy field and their oncolytic variants are currently under evaluation in many clinical trials. The cell killing properties of oncolytic adenoviruses against the cancer cells have been known for a long time. In addition, it is known that they can activate immune system. To achieve more selective and effective antitumor effects several modifications of oncolytic adenoviruses have been studied. During my internship I worked on the development of a new cancer vaccine platform based on peptide-coated conditionally replicating adenovirus (PeptiCRAd). The PeptiCRAd technology consists of a serotype 5 adenoviruses which are coated with tumorderived peptides. The aim of the thesis was to evaluate the antitumor efficacy of the PeptiCRAd. The cytopathic effects of the PeptiCRAd were studied in vitro using human adenocarcinoma cell line, A549. In this experiment three different treatments were used to study the cytopathic effects of the PeptiCRAd and Ad5Δ24-CpG- virus or polyK-SIINFEKL- peptide alone. The cell viability was assayed using MTS reagent and quantified by spectrophotometer. The antitumor effects were also studied in vivo using immunocompetent C57BL/6 mice bearing B16-OVA melanoma tumors. Tumor-bearing mice were treated with Ad5Δ24-CpG- virus, SIINFEKL- peptide or the PeptiCRAd. To evaluate the antitumor effects, tumor volume was observed after the treatments. In this study, I show that PeptiCRAd and Ad5Δ24-CpG- virus both have oncolytic effects in vitro against A549 cells. In vitro Ad5Δ24-CpG- virus showed significantly better cytopathic effects at high concentration compared to PeptiCRAd. In vivo the PeptiCRAd showed strongest antitumor effect on the growth of established tumors. At the end of the experiment the volume of the tumor was significantly smallest in the PeptiCRAd group.
  • Li, Mingchen (2021)
    Parkinson’s disease (PD) is a progressive chronic neurodegenerative disorder, which results in the selective loss of dopaminergic neurons in the substantia nigra (SN). The loss of these neurons results in the dysfunction of the nigrostriatal pathway bringing forth the characteristic motor symptoms seen in PD: postural instability, rigidity, slowness of movement and resting tremors. Non-motor symptoms, such as cognitive deficits, depression and impaired olfaction, typically emerge before motor symptoms. Currently available treatments only provide symptomatic relief with diminishing returns over time and no improvements on the overall outcome of the disease. Neurotrophic factors (NTF) have been of particular interest as a possible curative treatment for PD due to their potential for neuroprotection and neurorestoration. Mesencephalic astrocyte-derived neurotrophic factor (MANF) is an NTF that has shown promising results in numerous in vitro and in vivo studies of PD. However, therapy with MANF and other NTFs involves surgical intervention for local administration, as NTFs cannot cross the blood-brain barrier (BBB). Therefore, the therapeutic potential of a systemically administered NTF would be tremendous, as it would lead to a significantly more favorable risk-benefit ratio for the patient. The aim of the current investigation is to evaluate the efficacy of a next generation variant of MANF in the 6-hydroxydopamine toxin-induced unilateral lesion rat model of PD. Prior in vivo results suggested that subcutaneously injected MANF variant is able to penetrate the BBB. Amphetamine-induced rotational behavior (AMPH-ROTO) was used to evaluate the severity of the unilateral lesions during the experiment every other week until the end of the experiment at week eight. Animals were divided into treatment groups during week two based on their AMPH-ROTO results. Animals received MANF variant either subcutaneously through an implanted osmotic minipump at two different dosages or as a single dose divided into three separate intrastriatal injections. Tyrosine hydroxylase (TH) immunohistochemical staining was performed on brain sections collected from the striatum and SN for data analysis. In addition to AMPH-ROTO results, the efficacy of treatment was determined via the optical density of TH-positive striatal fibers and the number of TH-positive cells in the SN. Statistically significant differences (defined by p < 0.05 and a non-zero mean difference at a 95 % confidence interval) were observed only in the number of TH-positive cells in the SN favoring intrastriatal MANF variant treatment over both intrastriatal MANF and the vehicle treatment. The main concern regarding the validity of the results was related to the heterogeneous lesion sizes in different treatment groups possibly resulting in unsuccessful randomization due to excessive baseline differences. The inadvertent negative effects of this was further exacerbated by low a priori statistical power, which in the end had likely caused inflated effect sizes. Thus, assessment of the definitions of the used statistical parameters and the limitations of the experimental design suggest that presently, the efficacy of the MANF variant could not be evaluated reliably, in spite of the statistically significant result.
  • Pihlaja, Tea (2017)
    Cytochrome P450 (CYP) enzymes are important catalysers in the first phase of drug metabolism. Roughly two thirds of drugs are oxidized via CYP enzymes, which enable the further modification of drugs, and their excretion. In this thesis, human liver microsomes containing the main hepatic CYP enzymes were immobilized on thiol-ene based micropillar arrays and their stability was evaluated using a CYP2C9 isoenzyme specific luminescent substrate, Luciferin-H. The aim of the study was to develop microfluidic immobilized enzyme reactors (IMERs) for studying enzyme kinetics and drug-drug interactions. For this purpose, the instability issues associated with previously reported CYP-IMERs were carefully addressed. The CYP immobilization protocol used was based on a protocol previously developed in the context of other research projects and relied on biotinylation of human liver microsomes (HLM) with help of fusogenic liposomes. The biotinylated HLMs were then attached to the streptavidin-modified thiol-ene surfaces. The CYP activity was determined by utilizing microfluidics under continuous flow conditions (typically 5 μL/min) in the presence of NADPH. The luminescent metabolite formed by the CYP2C9 enzymes was quantified with a commercial well-plate reader from fractions collected at the microreactor outlet. Half-life was used to compare the differences between enzyme stabilities reached via different immobilization conditions. The effects of flow rate and reaction temperature on the stability of the CYP-IMERs was evaluated together with addition of antioxidative agents and reactive oxygen species (ROS) scavengers. Different functionalization steps as well as storage time and conditions were studied. With Luciferin-H as the model substrate of CYP2C9, the CYP-IMERs showed higher activity and stability at room temperature than at +37 °C. The peak activity could be increased via optimization of the immobilization protocol, though long-term storage diminished the peak activity. The activity of the IMERs typically attenuated within 1-2 hours with little or no improvement achieved via optimization of the immobilization or operation conditions. Only upon addition of the ROS scavengers, the peak activity and stability of the CYP-IMERs could be slightly improved. After functionalization, the IMERs maintained their activity until the time of use when stored in +4 °C for up to 2 weeks, but re-use of IMERs was not possible.
  • Nykänen, Tina (2013)
    Rhazya stricta Decne. is a small evergreen shrub belonging to the Apocynaceae family. The plant grows in South Asia and the Middle East, and in these areas it is used in traditional medicine. All parts of the plant are used in different preparations for a variety of purposes such as infections, bowel diseases, itching and diabetes. R. stricta synthesizes about a hundred different alkaloids, of which only a fraction has been studied closer. Some of the analyzed alkaloids have showed some interesting pharmacological properties such as antibacterial and cytotoxic properties. Because it is often both economically and ecologically unsustainable to cultivate or to collect large amounts of medicinal plants from nature, cell cultures have been developed from plants. The properties and synthesized substances of the cell cultures can be analysed and modified in laboratories. In the experimental part of this work, a system was developed for alkaloid extraction, fractionation and isolation from dried cell material from cultured R. stricta hairy root-cells. The goal was to develop a functioning system that eventually enables identification of the alkaloids synthesized by the cultured cells under given conditions. Alkaloids were extracted from 26 g of dried and ground cell mass. The fractionation of the alkaloids was performed with medium pressure liquid chromatography (MPLC) and the fractions were analyzed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The alkaloids were purified by horizontal TLC and preparative HPLC. Ion-pair chromatography was used for analyzing the extract, fractions and purified alkaloids. Five components from two fractions were eventually isolated. One of the components was tentatively identified as vincanine, but further analyzes have to be performed to identify all components reliably. In total, hairy root-cells seem to synthesize approximately 20 alkaloids with variable polarity.
  • Muurman, Tuulikki (2021)
    Background: Poor health literacy (HL) is associated to increased hospitalization and decreased seeking for screenings. Shared decision making can increase patient knowledge, decrease anxiety over the care process, improve health outcomes and reduce health care costs. Little is known about factors influencing health literacy and participation in treatment decision making in different population groups. Objectives: To investigate factors predicting HL and participation in the treatment decision making. Methods: A cross-sectional population online survey conducted in Finland in 2019 by Finnish Medicines Agency. Both health literacy and participation in the decision making were assessed by three statements that sum variables were created with score 1-5 (Cronbach’s alpha value 0.584 and 0.810). Age, gender, education, household income and most common chronic diseases were chosen as possible predicting factors. Two-variable Pearson’s chi-squared test was first used to find significant factors followed by logistic regression analysis to take into account several variables. Results: Of all the respondents (n=2104) 76.5% had good HL and 73.4% had willingness to participate in the treatment decision making. In the two-variable test older age (p<0.001), lower education (p<0.001), lower household income (p=0.001), higher number of chronic diseases (p=0.003), having cardiovascular diseases (p=0.003), diabetes (p=0.029) and cancer (p=0.001) predicted poorer health literacy. Male gender (p=0.001), not having chronic diseases (p=0.001), not having a musculoskeletal disorder (p=0.050) or mental health disorders (p<0.001) predicted poorer participation in the treatment decision making. In the logistic regression analysis older age and having cancer predicted poorer health literacy. Male gender and not having mental health disorders predicted less willingness to participate in the decision making. Conclusions: Older age and cancer predicts poorer health literacy and male gender poorer willingness to participate in the decision making. Further research should focus on investigating more in detail the contributing factors to these findings, and how health literacy in elderly and men’s involvement to the decision making could be improved.
  • Ylinen, Varpu (2016)
    Chronic myeloid leukemia (CML) is a malignant hematologic disorder, which is fatal without a treatment. Oral tyrosine kinase inhibitors (TKIs) have revolutionized the treatment of CML and transformed the disease to a chronic condition that can be treated at patient's home. The common problem in the treatment of CML is patient's poor adherence to TKIs. The regular, consistent use of TKIs is crucial to keep disease under control. For this reason and to obtain an optimal treatment outcome, adherence to TKIs is extremely important. The aim of the study was to assess reasons for poor adherence to TKI-medications in Finnish CMLpatients, including patient characteristics, treatment related factors, comorbidities and concomitant medications. In addition, patients' experiences, beliefs, knowledge and perception about CML and its treatment were explored and how these could contribute to nonadherent behaviour. This study is part of the larger study, assessing adherence to TKI treatment among Finnish CML population. The data was obtained by using patient questionnaires and semi-structured theme-interview during patient meetings in 2012. Study population consisted of Finnish adult CML patients who had been on TKI -medication (imatinib, nilotinib or dasatinib) for more than six months prior to the study baseline. Patients' adherence was measured using Morisky Medication Adherence 8-Item Scale (MMAS-8) and based on their score, patients were divided into three groups: high, medium and low adherence. Both quantitative and qualitative methods were used in data analysis. Study findings show that 21% (n=18) of the patients were low adherent and 23% (n=20) were high adherent to their treatment. Patient sociodemographic characteristics or experienced adverse drug reactions (ADRs) did not predict adherence, while more concomitant medications and comorbidities were associated with high adherence. However, ADRs had negative effect on the quality of life of several nonadherent patients. All nonadherent patients reported unintentional nonadherence and the most common reason was forgetting. Two-thirds of the patients (n=12) reported intentional nonadherence, which often was a result of experienced ADRs. The knowledge of CML and its treatment was poor among all patients while over half of the nonadherent patients (n=11) thought that they received enough information received. Overall, patients were very satisfied with care provided by the hospitals, physicians and other healthcare professionals. Managing TKI-treatment regimen is challenging for many patients and ADRs can have a negative impact on the quality of life. Healthcare professionals should regularly assess patient adherence and provide information and support for the patients to help them to succeed in medication management. Reasons for poor adherence are complex and have to be identified from each individual patient so that adherence can be improved.
  • Ritamäki, Kaisu (2019)
    Pharmaceutical companies are required to comply with fair market guidelines and regulations. However, definition of fair market value (FMV) in clinical trial is not unambiguous. In literature are some suggestions how to determine the phenomenon of FMV in clinical trial. Understanding the FMV and how it should be applied into practice when conducting clinical research is challenging. This study provides more focused information on FMV in clinical trials and its determination. FMV should be determined for research-related activities in clinical drug research. FMV of research related activities can be consistent if similar sites are performing similarly conducted studies for similar sponsors. Therapeutic area and geographical location of the trial site can also influence for the FMV. This study was performed in co-operation with Roche. The aim of the study was create a consistent and transparent method to assist in the determination of FMV in medical drug research in relation to the payments paid by the sponsor to the sites. Clinical trial agreements (CTA) and associated agreements were analysed to investigate FMV of research-related activities by study site, study type, therapeutic area and geographical area. Average price and price range of each research-related activity from previous CTAs and associated agreements of Roche Finland was calculated. Based on available data from literature and study results research-related activities and factors affecting to the FMV of clinical trials were discussed to create comprehensive understanding of FMV in clinical drug research. Based on this study average price of the specific research-related activities can be different by therapy area, site, study type and geographical area. All these factors are relevant when assessing FMV of specific research-related activity. Studied therapy area and site seems to have the most important impact when evaluating FMV. For some research-related activities such as national coordinator investigator (NCI) fee price ranges could be very big whereas in other research-related activities such as pharmacy fees prices could be quite similar. Some research-related activities were very study specific which affected evaluation of those activities. CTAs and associated agreements are valid documents to gather information assessing FMV of research-related activities in medical drug research. Average price and price range of the research related activity can be used when assessing FMV in medical drug research. However, price of the specific research-related activity need to be evaluated considering the studied therapy area, site, study type and geographical area.