Farmasian tiedekunta

 

Recent Submissions

  • Kanerva, Meeri (Helsingin yliopisto, 2019)
    Breast cancer is the most common cancer among women world wide and it´s incidence is constantly growing. The prognosis of local breast cancer is good and patients with metastatic breast cancer are living longer with their disease. The growing survivorship and population of chronically ill breast cancer patients has made quality of life one of the most important aspects in the treatment of breast cancer. Cytotoxic chemotherapy is a widely used treatment for breast cancer. Chemotherapy can cause difficult adverse events, which can affect the patients’ quality of life. Chemotherapy can also relieve the symptoms caused by cancer when used to treat metastatic breast cancer. The aim of this systematic review was to collect the currently available literature about breast cancer patients´ health related quality of life as comprehensively as possible, review the quality of the literature and the effects of chemotherapy on breast cancer patients ‘quality of life. The literature search produced 1666 references. According to the inclusion and exclusion criteria, 107 full text articles were accepted to the final systematic review, 53 of which reported the health related quality of life during adjuvant treatment of breast cancer, and 51 of which reported it during the treatment of advanced or metastatic breast cancer. In addition 3 previous systematic reviews were found. The basic information about the articles was extracted into a table. Articles were heterogeneous regarding their study settings, used quality of life instruments and reporting. Most studies used a disease specific quality of life instrument. The collected literature gave a strong indication of quality of life worsening during adjuvant chemotherapy of breast cancer. This observation was further supported by the previous systematic reviews. Most of the studies reporting the quality of life during chemotherapy for metastatic breast cancer, reported less than clinically important changes during the treatment. A few studies reported clinically important worsening or improvement in quality of life. 11 studies, which were made during or after 21: st century, which reported numerical data from quality of life, which reported predominantly quality of life and which had sample size of at least 100 patients in baseline, were accepted to further assessment of quality of the studies and closer observation. The quality of the studies was assessed with STROBE and CONSORT checklists. The quality of studies was heterogeneous as the studies fulfilled 44.8 % to 86.1 % of the scoring items. Only one randomized controlled trial reported quality of life as their primary end point. The data from these studies supported the previous observation of quality of life worsening during adjuvant chemotherapy of breast cancer. The effect of chemotherapy during metastatic breast cancer on quality of life was not unambiguous. Both clinically meaningful worsening and improvement of quality of life was reported. Breast cancer patients´ health related quality of life has been assessed in multiple publications, but the existing literature is heterogeneous and it´s use in decision making and economic evaluation is not easily feasible. Breast cancer patients´ health related quality of life worsened during adjuvant chemotherapy. Significant improvement in breast cancer patients´ health related quality of life was not observed during chemotherapy for metastatic breast cancer.
  • Karasti, Eveliina (Helsingin yliopisto, 2019)
    The amount of informal caregiving has increased in Finland, with a growing emphasis on the older adults. Although the medication management process in informal caregiving has been studied and is known to have significant risks, research data focusing on the older adults is still limited. The aim of this study was to describe the medication management process of informal carers and care recipients of at least 65 years old. The aim was to identify medication errors and medication risks in the medication management process and to find out how the caregivers manage them. In addition, the study examined the informal carers and care recipients own development proposals to improve medication management process. A total of 21 volunteer informal carers and care recipients living in the Helsinki metropolitan area were recruited to this study. The study was conducted as a qualitative interview survey in the homes of the participants. The interviews were a combination of semi-structured interviews and narrative approach. The material to this study was collected during spring and summer 2018. The study was analysed with abductive content analysis combining both deductive and inductive approach. The aim was to find repetitive elements by encoding and grouping expressions. The results of this study were compared with previous theory and the results were supplemented with a picture of the medication management process and a fishbone diagram was drawn from the risk factors and contributing factors of the medication management process. The families had medication errors in all stages of the medication management process. The most frequent medication errors were found in counselling, medication administration and in medication treatment monitoring. As a contributing factor, the healthcare professionals’ haste and the responsibility of the caregiver recurred in the background of the medication errors. Family caregivers and care recipients wish to have more counselling, more monitoring of medication and better interaction with health care. Carers often felt left alone to take care of another person's medications and felt they lack support from healthcare. By facilitating access to a physician, improving the availability of a physician, increasing the amount of counseling and support provided by healthcare, including pharmacies, could the safety of medication management at home be improved.
  • Äijö, Nelli (Helsingin yliopisto, 2019)
    As the population becomes older and the amount of multimorbid patients increases, also health care spending increases. New care models are needed where patients’ needs are taken into consideration by providing preventive and patient-centred care. In Finland and internationally, new ways to treat elderly, chronically ill patients have been developed. One of the new models is the health and care plan model. This longitudinal, randomised, controlled trial studied the health and care plan model’s impact on healthcare costs, patients’ physical functioning and patients’ quality of life. The aim of the study was to study the cost-effectiveness of the health and care plan model compared to standard care practice. The goal of this study was to study if rational pharmacotherapy and self-management support can prevent the decline in physical functioning, the decline in quality of life and the increase in health service use and costs among elderly population. This study was conducted between 2014-2018 as a multi-disciplinary cooperation between Tornio health station, University of Helsinki Faculty of Pharmacy and Alatornio pharmacy. The patients in this study were over 75-year old, multimorbid, community dwelling, polypharmacy patients. The patients in the intervention group received an interview based clinical medication review and were formed a medication plan. Furthermore, the patients in the intervention group were planned a health and care plan that was combined with the medication plan into a comprehensive self-management plan in a multi-disciplinary meeting. A case manager was appointed for the patients in the intervention group and the case manager could be contacted by the patients in the intervention group at any point of the study if there arose a non-acute concern with the patient’s health. The patients in the control group were conducted a prescription review based on the information available in the electronic health records system and continued receiving the standard care practice. Cost and effectiveness data were gathered from the patients over the period of two years. The effectiveness data were gathered at Tornio health station where the intervention and control group’s quality of life was measured with the SF-36 generic quality of life measure and physical functioning was measured with Short Physical Performance Battery (SPPB). The quality of life data gathered with SF-36 were transformed into one preference based single index score SF-6D to calculate the quality of life and quality adjusted life years (QALY). Data about the use of health services were extracted from the electronic health records system and transformed into costs by using the national reference costs. At the beginning of the study, the intervention and control group were statistically significantly similar. During the two-year follow-up, no statistically or clinically significant differences were observed between the intervention and control group in their quality of life, in their physical functioning or in the costs of used health services. However, in the intervention group, the cost of used health services was on average 2 406 euros smaller than those of the control group’s during the two-year follow-up. The health and care plan model was cost-effective compared to standard care practice. The incremental cost-effectiveness ratio was -64 504 € per one QALY. Based on this study, it is recommended to support the self-management and physical functioning of the elderly with an intervention like health and care plan model to decrease the health care spending. The results of this study can be applied to Finnish healthcare system to decrease the health care spending of multimorbid, community dwelling and polypharmacy elderly patients. The use of real-world evidence increases the reliability of this study.
  • Vainio, Sanna (Helsingin yliopisto, 2019)
    Despite the long history of skin grafting, there is no standardized treatment for split-thickness skin graft donor sites. These sites cause a notable amount of pain and discomfort to the patients and open wounds also introduce a risk for infection. There is an extensive need for treatment options promoting the fastest and least painful healing possible while also being infection-free. The treatment of split-thickness skin graft donor sites is constantly studied and there is plenty of scientific literature available about this topic. In the theory section of this Master’s thesis, the structure of skin, the process of wound healing, skin grafting surgery and wound care products for split-thickness skin graft donor sites are briefly introduced. Additionally, the method of systematic review is described. In the empirical section, a systematic review is performed to compare animal- and non-animal-based wound care products in the treatment of split skin graft donor sites. The methodological quality of the included studies is reviewed. In the literature search, 3552 references were found. In this systematic review a total of 23 articles were included comprising of 21 comparative clinical studies and two previous literature reviews. Of the original studies, 20 reviewed healing, 14 infection and 17 pain of the split-thickness skin graft donor sites. Based on the results of the systematic review, animal-based wound care products might promote healing and reduce pain experienced by patients in the treatment of split-thickness skin graft donor sites when compared with non-animal-based wound care products. The results concerning infection were inconsistent. Generally, the reporting of the clinical original studies was not comprehensive enough for proper evaluation of methodological quality. Some defects, mostly in the blinding of the patients, study personnel and the assessors of outcomes, were also found. Moreover, the studies were heterogeneous in their definitions and measuring of the reported outcomes. Therefore, there is substantial uncertainty in the results of this systematic review. The systematic and transparent way of conducting the literature search, the review of the methodological quality and the reporting of the outcomes can be considered as a strength of this thesis. The main weakness is, that only one person performed the critical steps of this study, which might increase the risk of bias and reduce the repeatability of the study.
  • Aalto, Aura (Helsingin yliopisto, 2019)
    Lääkevalmisteiden, lääkinnällisten laitteiden ja sekä lääke- että laiteosan sisältävien yhdistelmätuotteiden tulee täyttää niiden tehoa, turvallisuutta ja laatua koskevat viranomaisvaatimukset ennen markkinoille tuloa. Lääkkeiden ja lääkinnällisten laitteiden rekisteröinnissä dokumentaatiovaatimukset, hakemusprosessit ja viranomaiskäsittelyt eroavat kuitenkin toisistaan. Yhdistelmätuotteet rekisteröidään Euroopan unionissa joko lääkkeinä tai laitteina niiden pääasiallisen vaikutusmekanismin mukaan. Laitteiden rekisteröintivaatimukset ovat parhaillaan muuttumassa, sillä uusi lääkinnällisiä laitteita koskeva EU-asetus 2017/745 (medical device regulation, MDR) astui voimaan toukokuussa 2017. Asetuksen käyttöönoton siirtymäkausi jatkuu toukokuulle 2020 asti. Tutkimus on jatkoa aiemmin toteutetulle tutkimukselle (Nuolimo 2016), jossa selvitettiin lääkkeiden ja lääkinnällisten laitteiden rekisteröintiprosessien eroja. Aiemman tutkimuksen johtopäätöksenä oli, että rekisteröintiprosessien erot vaikuttaisivat suosivan valmisteen rekisteröintiä CE-merkityksi terveydenhuollon laitteeksi sellaisissa rajatapaustilanteissa, joissa rekisteröinti sekä lääkevalmisteeksi että lääkinnälliseksi laitteeksi on mahdollista. Nyt tehdyn teemahaastattelututkimuksen tavoitteena oli edelleen selvittää rekisteröintiprosessien eroja ja erityisesti tuotteen rekisteröintistatuksen valintaan vaikuttavia taustatekijöitä. Tutkimuksen tavoitteina oli myös selvittää, miten yhdistelmätuotteiden rekisteröintiprosessi eroaa muista rekisteröintiprosesseista ja miten uusi lääkinnällisiä laitteita koskeva EU-asetus 2017/745 vaikuttaa käytännön rekisteröintityöhön. Teemahaastattelututkimuksessa haastateltiin kunkin tuoteryhmän rekisteröinnin asiantuntijoita. Tutkimukseen osallistui yhteensä kahdeksan haastateltavaa. Rekisteröinnin asiantuntijoiden työtoimenkuvat vaihtelivat niin paljon, että rekisteröintiprosessien vertailu oli vaikeaa. Tutkimuksessa todettiin, että laitteiden rekisteröintiprosessi on kevyempi kuin lääkkeiden. Rekisteröintistatus valitaan tuotekohtaisesti. Yhdistelmätuotteiden rekisteröintiprosessin raskaus riippuu siitä, onko tuote rekisteröity lääkkeeksi vai laitteeksi. Yhdistelmätuotteiden kohdalla rajanvetotilanteet, joissa täytyy määritellä, rekisteröidäänkö tuote lääkkeeksi vai laitteeksi, voivat olla haastavia. Uuden laiteasetuksen implementointi oli tutkimusajankohtana vielä kesken, mutta asetus oli jo lisännyt työtä laite- ja yhdistelmätuotteiden rekisteröinnissä ja tuonut uusia liiketoimintamahdollisuuksia CRO-yrityksille.
  • Häggman, Verner (Helsingin yliopisto, 2019)
    If pharmaceutical quality system fails it causes a hazard to the patient’s health, but also to the manufacturer’s economy. For this reason, the manufacturer’s must make sure their products comply with the quality requirements placed by authorities. To ensure the compliance, the authorities perform inspections at the manufacturing sites. If the site does not comply with the quality requirements, the authority will take necessary measures. The goal of this study was to find what type of quality issues FDA and the authorities within EU have observed while inspecting manufacturing facilities, which of these issues are most common, in which countries the sites companies with issues have been located. The results were assessed from European pharmaceutical company’s point of view. The data for the study was collected from Eudra GMDP database and from FDA Warning letters sent by FDA headquarters from years 2015-2017. Qualitative analysis of content was chosen as the method of analysis. The collected data was classified into main classes and subclasses based on reoccurring topics. The classes were transferred in tables to compare how which of the classes were most common. Most often the facilities with quality issues were located in China and India. The authorities also perform a lot of quality inspections in these countries, but that alone doesn’t explain the large number of quality issues in these countries. The number of sites with quality issues per inspection was also high. Both the authorities of EU countries and FDA had mainly observed similar issues. Often quality issues were related to data integrity. Other common themes were quality management system, cleaning of equipment and facilities and analytical methods. There were also some differences in the observed issues. E.g. FDA had rarely observed issues related to personnel while EU authorities had observed such issues frequently. Quality issues which had led to measures by authorities were often related to larger problems with the quality management or to very basic quality actions. If company doesn’t have well-functioning quality organization, the quality system is often inadequate also in other ways. By comparing their own activities with the issues observed at other companies, it is easier for a company to improve their quality and avoid major quality issuer before they occur.
  • Niklander, Johanna (Helsingin yliopisto, 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.
  • Viljakainen, Tuulikki (Helsingin yliopisto, 2019)
    Parkinson’s disease is a progressive neurodegenerative disease, in which dopamine neurons are dying in the nigrostriatal dopaminergic pathway. This causes motor symptoms such as slowness of movement, tremor, and rigidity. In addition, various non-motor symptoms appear. All currently used medicines are symptomatic, and there are no disease modifying treatment available for Parkinson’s disease. Several neurotrophic factors have shown promise in animal models of Parkinson’s disease. One of those is cerebral dopamine neurotrophic factor (CDNF) which has been studied in different animal models, including rodents and non-human primates. CDNF is a secreted protein but it is also localized in endoplasmic reticulum (ER). CDNF has two domains, N-terminal and C-terminal, which may have distinct functions. CDNF can be retained in the ER by the ER retention sequence at the end of the C-terminal domain. The C-terminal domain also has an evolutionarily conserved disulfide bridge which is crucial for the biological activity of CDNF. The exact mechanism of CDNF is still unknown. However, it has been shown that CDNF affects the unfolded protein response (UPR) in the presence of ER stress. Neurotrophic factors do not penetrate blood-brain barrier (BBB), for this reason, they need to be injected directly to the brain. Penetration of the BBB is also a problem in the treatment of many other diseases. Various methods for enhancing the BBB penetration of drugs have been studied. For example, permeability of the BBB can be temporarily increased by focused ultrasound combined with microbubbles. Another possibility is the use of a carrier molecule, which can be transported through BBB via specific transport mechanisms. Furthermore, molecule modification offers many applications to achieve enhanced BBB penetration. In view of peripheral administration, a next generation variant of CDNF (ngCDNF) has been developed. The efficacy of this novel variant after intrastriatal injection is equal to that of CDNF in a 6-hydroxydopamine (6-OHDA) rat model of Parkinson’s disease. Systemic administration could also enable treatment of non-motor symptoms of Parkinson’s disease. The aim of this experiment was to study the effects of subcutaneously injected ngCDNF on rotation behaviour, and nigrostriatal TH-positive cells in rats with 6-OHDA lesions. 6-OHDA was injected unilaterally to three different sites in the striatum. Two weeks later, the lesion size was estimated, via amphetamine- induced rotation test. ngCDNF, at two dose levels, was injected twice weekly for three weeks. Amphetamine-induced rotation test was assessed every other week, until week 12. At the end, optical density of tyrosine hydroxylase (TH) was measured from sections of the striatum, and TH positive cells in the substantia nigra were counted. In addition, the effect of ngCDNF on anxiety and depression like behaviour, learning, and locomotor activity were studied at three different levels in naïve mice. Behaviour was analyzed by open field test, forced swim test, and fear conditioning test. The ngCDNF did not seem to have clear effect on rats’ behaviour or TH positive cells and fibers compared to the control group, but positive tendency was found in the group with lower dose. The reduced efficacy of ngCDNF,via subcutaneous administration, is likely due to rapid metabolism and insufficient entry of the active form to the brain. In naïve mice, ngCDNF did not reduce anxiety-like behaviour and did not affect locomotor activity after subcutaneous injections. This result supports previous findings, which suggest that the effects of CDNF are specific to the toxin treated cells and CDNF has no effect in naïve animals.
  • Skullbacka, Simone (Helsingin yliopisto, 2019)
    Many drugs are associated with the risk of QT prolongation and torsades de pointes (TdP). The risk increases with other risks factors for QT prolongation. Recognizing risk factors and QT prolonging drugs is critical in the management of this drug-related problem. The aim of this master’s thesis was to study the prevalence of use of QT prolonging drugs in older adults receiving home care. Additionally, the aim was to study concomitant use of QT prolonging drugs as well as clinically significant QT prolonging drug-drug interactions in the participants. The secondary objective was to study the most commonly used QT prolonging in the participants. The material used in this master’s thesis originated from a randomized controlled trial in City of Lohja, Finland, which enhanced a coordination in medication risk management for older home care clients. The analysis of the baseline data collected in fall 2015 was only deepened regarding QT prolonging drugs. The participants (n=188) were older adults (≥65 years) receiving regular home care from City of Lohja, randomized into an intervention group (n=101) and a control group (n=87). The majority of the participants were women (69%). The mean age of the participants was 83 years. Data on the participants’ drugs were collected from their medication lists. Clinically significant drug-drug interactions were identified using the SFINX database. The QTDrugs Lists of CredibleMeds were used for identifying drugs associated with QT prolongation and TdP. On average, the participants (n=188) used 2.3 drugs (SD 1.3, median 2.0) associated with QT prolongation and TdP. Of the participants, 36% (n=67) used drugs with known risk of TdP (QTDrugs List 1). The most commonly used drugs with known risk of TdP were donepezil and citalopram. The prevalence of QTDrugs List 2 drugs (possible risk of TdP) was 36% (n=67). Most of the participants (n=156, 83%) used drugs which under certain circumstances are associated with TdP (QTDrugs List 3). One fifth (21%) of the participants used concomitantly 2-3 drugs associated with QT prolongation and TdP. QT prolonging drugdrug interactions (SFINX-D interactions) were found in 3% of the participants. The drugs involved in the drug-drug interactions were donepezil, (es)citalopram and haloperidol. The prevalence of use of clinically relevant QT prolonging drugs (QTDrugs Lists 1-2) was higher in this study compared with the prevalence in outpatients in previous studies. Concomitant use of QT prolonging drugs is common in outpatients. Health care professionals need to be educated on the risks of QT prolongation, TdP and the risks of using QT prolonging drugs concomitantly. Risk assessment tools considering patient-specific risk factors could be more widely used, as they may reduce modifiable risk factors, and actual events of QT prolongation and TdP may be avoided. There is a need for systematic procedures for assessing and managing the risks of QT prolongation and TdP in the Finnish health care system.
  • Andersson, Ville (Helsingin yliopisto, 2019)
    The Finnish Medicines Agency, Fimea, is the authority responsible for supervision pharmacies in Finland. Recently, there has been more interest in Fimea to improve its supervision of community pharmacies. For this purpose, a questionnaire was made. Prior to the making of the questionnaire, community pharmacy supervision practices were studied in Nordic countries and in the UK. Additionally, faults found in Finnish community pharmacy inspections in 2016–2018 were classified by analysing anonymized fault lists (n=94) separated from inspection reports. When the most common faults were identified, it was possible to include questions concerning these faults into the questionnaire. A modified version of the Delphi method was used when developing the questionnaire. Comments on the applicability of the questions were given by a panel of experts consisting of inspectors of Fimea. The questionnaire was subsequently edited in accordance with the given comments. Separate versions of the questionnaire form were developed for community pharmacies and for their subsidiary pharmacies. At the end of this study, the questionnaire was sent to seven pharmacies and to three subsidiary pharmacies. After the results of the questionnaire were collected, Fimea gave feedback on the questionnaire. 25 categories were created by classifying faults found from pharmacy inspections. The most common inspection observations were faults in storage condition monitoring (97 % of pharmacies), narcotics (86 %), implementation of code of conduct (86 %), product errors (86 %) and preparation of medicines ready for use (81 %). The questionnaire begins by asking basic information about the pharmacy. Following questions concern the personnel and their further adequacy training. The questionnaire also includes several questions on the code of conduct within the pharmacy. Additionally, there are questions about storage condition monitoring, dispensary and accounting of narcotics. At the end of the questionnaire, there are also a few questions about the European Medicines Verification System (EMVS) which will be implemented by February 2019. Support from the inspectors of Fimea and studying regulations of pharmacies helped identify appropriate questions for the questionnaire. However, the perspective of the questionnaire may be limited due to the questionnaire being developed based up on faults found from inspections. The faults observed from inspections across pharmacies in Finland have been very similar with some of them being also alarmingly common. Because many of the observed faults are relatively easy to fix, simple corrective measures could be implemented to improve the situation across several pharmacies. Thus, usage of questionnaires, such as one made in this study, could be considered a feasible way of improving supervision of pharmacies.
  • Neuvonen, Janina (Helsingin yliopisto, 2019)
    Flowability of powders is in critical role when manufacturing the most popular dosage forms, tablets and capsules, of pharmaceutical industry. Re-formulation is expensive and time-consuming, so it is important to determine powder flow properties at the initial stage of drug development prior to tabletting and encapsulation processes. There are many different methods, like shear cell, flow through an orifice and bulk and tapped density, to examine powder flowability. Despite the methods, the most reliable means of examining powder flowability is often empirical. In early stages of drug development, it would be good to have faster, more reliable and cheaper methods to examine powder flowability. FT4 Powder Rheometer is a relatively new flowability characterization technique. The aim of this study is to find out whether the library created using the FT4 Powder Rheometer methods makes it possible to characterize the rheological properties of solids in the early stages of drug development. In addition, the aim is to investigate whether FT4 Powder Rheometer methods can predict the success of masses in tableting and encapsulation processes. The information gained from the research can be used in the future, for example, in continuous processes, because flowability plays an important role, especially in the supply of raw materials to the process, which is the most important division of continuous processes. To the library were selected for particle size and shape 15 different types of material. These materials were subjected to five different FT4 Powder Rheometer basic test methods. In addition, the particle size and shape of the materials and the flow through an orifice and the bulk and tapped density were determined to support the results of the powder rheometer. The principal component analysis was used to process the results. As the tablet and capsule masses examined, the masses of a previous study were utilized. Those masses were tableted and encapsulated in that previous study. These tablet and capsule masses contain a variable amount of cohesive drug substance. FT4 Powder Rheometer methods provide more complex information about materials and their behaviour than conventional flowability test methods. From the powder rheometer parameters pressure drop, compressibility and specific energy distinguish the cohesive and the non-cohesive materials, because the cohesive materials with these parameters obtain clearly higher values than non-cohesive materials. Additionally, the cohesion of FT4 Powder Rheometer shear cell test mainly distinguishes highly cohesive materials from other materials. The flow rate index makes it possible to separate the materials to which the change in flow rate particularly affects. Fluidizing materials, due to the air flow, are distinguished by the aeration test. Avicel PH-102 could be used as a rough limit value for well and poorly flowing materials in the created library (excluding the aeration and shear cell test). Stability index -, flow rate index -, specific energy -, pressure drop -, and compressibility-results of the FT4 Powder Rheometer correlated to the proportional proportion of the cohesive drug in the mixture. These parameters could possibly be used to distinguish mixtures containing the cohesive material. Additionally, specific energy, compressibility, pressure drop, basic flowability energy, stability index and flow rate index correlated with the weight variation of the tablets. With these parameters one could possibly assess the tabletability of the mixtures. A much larger library is needed to evaluate and predict the rheological properties of new materials. FT4 Powder Rheometer can possibly be used to predict the tableting success of tablet and capsule masses. This would be interesting to look more extensively, for example as part of a library. Additionally, it would be good to investigate whether the results of powder rheometer correlate to continuous production.
  • Mäkinen, Arttu (Helsingin yliopisto, 2018)
    This is a systematic review aiming to investigate the efficacy, effectiveness, and safety of biosimilars in the treatment of inflammatory bowel diseases. Biosimilar drugs used to treat inflammatory bowel diseases include biosimilar infliximab and biosimilar adalimumab. Biosimilar infliximab has been authorized by the European Medicines Agency (EMA) in 2013 and by the US Food and Drug Administration (FDA) in 2016. Biosimilar adalimumab has been authorized by EMA and FDA in 2017 and, at the time the literary search for this systematic review was conducted no studies were found regarding the treatment of adalimumab biosimilar for inflammatory bowel diseases. To acquire marketing authorization for biosimilars, it must be proven that the biosimilar is biologically similar to the original medicinal product. Bioequivalence is demonstrated through physicochemical trials and clinical trials. However, clinical trials do not have to be performed with all of the indications for which the original medical product is registered. After proving bioequivalence with one or more indication it is possible to extrapolate the biosimilar to be used in all of the original medical products indications. This has raised the question of whether biosimilars are really comparable to the originator in indications for which no clinical trials have been conducted. This systematic review was implemented using the Cochrane Handbook for Systematic Reviews and Interventions. Systematic literature searches were made in Cochrane, Medline (Ovid®), PubMed and Scopus databases on 12.05.2017. 14 observational studies, one systematic review and a randomized clinical trial that met the inclusion criteria were included in the systematic review. The quality of the publications was evaluated using the STROBE-, NOS- and CONSORT-checklists and information regarding the efficacy, effectiveness and safety of biosimilars was extracted. CD-patients receiving tumor necrosis factor alpha inhibitors for the first time, the clinical response was achieved in 50.0 % to 97.2 % of patients depending on patient population and the duration of treatment. Similarly, for UC-patients, the clinical response was achieved in 62.2 % to 100.0 %. The clinical remission was achieved among 28.9 % to 84.4 % of CD-patients and among 28.9 % to 84.4 % of UC-patients, depending on patient population and treatment follow-up. After the switch from original infliximab to biosimilar, the proportion of patients in clinical remission during follow-up ranged from 62.3 % to 100.0 % in CD-patients and from 45.5 % to 100.0 % in UC-patients. Clinical remission was sustained throughout the whole follow-up in 70 % to 100 % of CD-patients and 66.7 % to 92.0 % of UC-patients. The incidence of adverse events leading to the discontinuation of drug treatment was between 0.0 % and 25.0 %, and the incidence of all adverse events ranged from 0.0 % to 93.6 % in CD- and UC-patients. Biosimilar infliximab seems to be comparable to the original product regarding the efficacy, effectiveness and safety. This result is supported by the systematic literature review published earlier. Conducting a meta-analysis of the information contained in this systematic literature review could have led to a more final decision considering efficacy, effectiveness and safety of biosimilar-infliximab in the treatment of inflammatory bowel diseases.
  • Karhunen, Emilia (Helsingin yliopisto, 2018)
    Functional in vitro cultured human hepatocytes are needed in different applications in biomedical research. Treatment for liver diseases is usually liver transplantation, but due to the lack of healthy donors, cell therapy using hepatocytes is considered as a better option. Drug industry will also need representative liver models to test metabolic profiles of drug molecules. Primary human hepatocytes are studied in cell therapy and disease modelling, but they have also drawbacks. In vitro they do not proliferate efficiently, and they are short-lived. In vitro differentiated human pluripotent stem cells (hPSCs) to hepatic fate are an alternative for the primary human hepatocytes. Especially human induced pluripotent stem cells (hiPSCs) are widely studied because they are easily available, and they even make personalized therapy possible without problems with ethical issues related to the human embryonic stem cells (hESCs). Differentiation to hepatic fate includes several steps before mature functional hepatocyte-like cells are formed. Hepatocytes are derived from the definitive endoderm (DE) which is one of the germ layers formed in the gastrulation process. Efficient induction of hPSCs into DE lineage would be a good starting point for generating mature hepatocyte-like cells in further hepatic differentiation. Different protocols to differentiate hPSCs in vitro into DE have been published. In vitro cell culture systems should well represent the environment of the target tissue because signals from the environment guide the differentiation. Three-dimensional (3D) cell culture systems are widely studied, because they better mimic the in vivo microenvironment of cells than two-dimensional (2D) cell culture. The aim of the thesis was to study the efficacy of the 3D differentiation of hiPSCs into DE. Before starting the 3D differentiation, differentiation protocol was optimized and the effect of ROCK inhibitor Y-27632 was investigated. Differentiation medium was supplemented with Y-27632 during the whole 6 days differentiation, because survival of the cells and formation of the spheroids were improved, and gene expression studies of pluripotency markers and several DE markers did not show evident effect of Y-27632 on the gene expression of hiPSCs. The main objective in the studies was also to investigate possible differences between different 3D culture conditions on hiPSCs differentiation into DE. Also, the effect of the spheroid size on differentiation was examined. Two different hydrogels were used as a matrix material in the experiments: basement membrane extract (BME) and nanofibrillar cellulose (NFC) hydrogels. Suspension culture was used as a biomaterial-free 3D culture system. Experiments were performed with three spheroid sizes: 200 cells/spheroid, 500 cells/spheroid and 1000 cells/spheroid. Efficacy of differentiation to DE lineage was estimated by studying protein and mRNA expression of some of the DE markers (HNF3B, SOX17, CXCR4, CER1), pluripotency marker OCT4, mesendoderm marker Brachyury and hepatoblast marker HNF4A in the cells. Spheroids differentiated in suspension and NFC were analysed by flow cytometry to get the number of DE positive live cells and dead cells using CXCR4 and 7-AAD double staining. Besides flow cytometry, protein expression of some of the key markers were studied by immunofluorescent staining and further confocal imaging. Viability of the spheroids in BME hydrogel culture were investigated using live/dead staining followed by confocal imaging. BME hydrogel culture was left out from the further experiments due to the morphology of the spheroids and results from viability and protein expression studies. Spheroids in suspension started DE differentiation faster compared to NFC culture. Suspension and NFC cultures yielded high number of double positive cells in flow cytometry and bright fluorescence of other DE markers was seen in the confocal images. NFC hydrogel proved to be a promising 3D culture system by supporting the differentiation of hiPSCs. Flow cytometry results and gene expression studies propose that four days long 3D differentiation would be efficient to produce sufficient number of DE cells. Smaller spheroids showed higher number of DE positive cells than bigger spheroids on day 2 but gene expression studies showed difference in DE marker expression between size conditions rather in later days in differentiation and it was the opposite. Experiments showed signs of more efficient differentiation of the smaller sized spheroids in the beginning of differentiation. But further studies are needed to verify the obtained results and both draw conclusions about the possible differences between different 3D culture systems and explore the best size of the spheroid for hepatic differentiation. However, results obtained from the studies are useful for designing further experiments.
  • Bruun, Tanja (Helsingin yliopisto, 2018)
    Marine organisms can be regarded as a diverse source of bioactive compounds with the possibility to discover novel drug lead molecules. Sea sponges produce bromine containing alkaloids, bromotyrosines, from which several are active against cancer. Some bromotyrosines have spirocyclic structure and the innate three-dimensionality and structural novelty of spirocycles make them an interesting option in drug design. Clavatadine C, extracted from sponge Suberea clavata, is a bromine containing spirocyclohexa-dienylisoxazoline alkaloid. It’s symmetric spirocyclic core can be viewed as a restricted derivative of open chain oximes, such as purpurealidin I, a bromotyrosine extracted from Pseudoceratina purpurea. Earlier work with purpurealidin I derivatives against melanoma cell line has had some promising results. Inspired by these earlier results, eight spirocyclic clavatadine C derivatives were synthesized according the published synthesis route. The activities of seven synthesized clavatadine C derivatives were tested on A375 melanoma cell line. All spiro derivatives were active with CC50 values ranging between 1.0 μM and 3.4 μM. Also, the activities of 10 earlier synthesized bromotyrosine derivatives were tested, from which four open chain oximes had CC50 values between 13.5 μM and 27.8 μM. Interestingly, the most active compounds were chlorinated and unhalogenated spirocyclic derivatives. In general, the spirocyclic compounds were 2- to 8-fold more active than the corresponding open chain oximes. The selectivity of active compounds was determined as cytotoxicity against Hs27 fibroblasts and by comparing the CC50 values of these two cell lines. The most selective compound was brominated derivative which had three times better selectivity against melanoma cells. The weak selectivity was consistent with the trend with open chain oxime analogs. Despite the selectivity issue, the improved activity of spirocyclic derivatives are promising and support for further investigation of marine-based spirocyclic bromotyrosine derivatives against melanoma.
  • Savolainen Roosa (Helsingin yliopisto, 2018)
    Liposomes are nano-sized vesicles in which the aqueous phase is surrounded by lipid-derived bilayer. They are excellent drug vehicles for example in ocular drug delivery because they can, among other things, increase the bioavailability and stability of the drug molecules and reduce their toxicity. Liposomes are known to be safe to use, because they degrade within a certain period of time and they are biocompatible with the cells and tissues of the body. Owing to its structure, the surface of liposomes can also be easily modified and functionalized. Light-activated ICG liposomes allow drug release in a controlled manner at a given time and specific site. Their function is based on a small molecule called indocyanine green (ICG) which, after being exposed to laser light, absorbs light energy and thereby locally elevates the temperature of the lipid bilayer. As a result, the drug inside is released into the surroundings. The blood circulation time of liposomes has often been prolonged by coating the liposomes with polyethylene glycol (PEG). Although PEG is generally regarded as a safe and biocompatible polymer, it has been found to increase immunological reactions and PEG-specific antibodies upon repeated dosing. Conversely, hyaluronic acid (HA), is an endogenous polysaccharide, which is present in abundance for instance in vitreous. Thus, it could serve as a stealth coating material which extends the otherwise short half-life of liposomes. One of the main objectives of this thesis was to find out whether HA could be used to coat liposomes instead of PEG. In order to prepare HA-coated liposomes, one of the lipid bilayer phospholipids, DSPE, had to be first conjugated with HA. For the conjugation, potential synthesis protocols were sought from the literature. Ultimately two different reductive amination-based protocols were tested. Consequently, the protocol in which the conjugation was achieved via the aldehyde group of HA, proved to be working. Thereafter, HA-coated liposomes were prepared by thin film hydration from the newly synthesised conjugate as well as DPPC, DSPC and 18:0 Lyso PC. Calcein was encapsulated in the liposomes. HA-covered liposomes were then compared with uncoated and PEGylated liposomes by examining their phase transition temperatures, ICG absorbances, sizes, polydispersities, and both light and heat-induced drug releases. The aforementioned tests were also conducted when the effects of the HA and ICG doubling were examined and the possibility to manufacture HA liposomes with small size was assessed. HA-liposomes showed similar results as PEG-coated liposomes. In addition, successful extrusion of HA-liposomes through a 30 nm membrane was also demonstrated in the results. Doubling of HA did not significantly affect the results. In contrast, increasing the molar amount of ICG by double caused spontaneous calcein leakage even before any heat or light exposure. Based on these findings, HA could work as a coating material instead of PEG, yet further studies are required for ensuring this conclusion. The other key objective was to evaluate the stability of four different formulations, named as AL, AL18, AL16 and AL14, in storage and biological conditions. Based on the differences in the formulation phospholipid composition, the assumption was that AL would be the most stable of the group and that the stability would decrease so that AL18 and AL16 would be the next most stable and eventually AL14 would be the least stable formulation. As in the previous study, the liposomes were prepared by thin film hydration with calcein being encapsulated inside the liposomes. In the storage stability test, liposomes were stored in HEPES buffer at either 4 °C or at room temperature for one month. In the test conducted in physiological conditions, the liposomes were added either to porcine vitreous or fetal bovine serum (FBS) and the samples were incubated at 37 ºC for five days. Regardless of the experiment, phase transition temperatures as well as light and heat-induced drug releases were initially measured. As the test progressed, calcein release, ICG absorbance, size, and polydispersity were measured at each time point. The initial measurements confirmed the hypothesis about the stability differences of tested formulations. In the storage stability test, all formulations, except AL14, appeared to be stable throughout the study and no apparent differences between the formulations or temperatures were observed. On the other hand, the stability of liposomes stored in biological matrices varied so that the liposomes were more stable in vitreous than in FBS and the stability decreased in both media as expected.
  • Tamminen, Matti (Helsingin yliopisto, 2018)
    Membrane-bound pyrophosphatases (mPPases) are a potential target for drugs against many neglected protozoan diseases, such as malaria, leishmaniasis, toxoplasmosis and trypanosomiasis. New drugs against these diseases are urgently needed, as the clinically used ones are either not effective, suffer from side effects, or resistance against them is developing. The mPPases of these protozoans are genetically conserved, while mammalian DNA does not encode them. A drug development project to find mPPase inhibitors was started, based on mPPase structures solved through X-Ray crystallography. Four hit compounds were identified. The aim of this study was to investigate the binding of these hit compounds at the mPPase binding site, and based on these results, to develop and synthesize novel compounds with higher affinity. A hit compound with an isoxazole ring was chosen as the model compound to be developed further. These novel compounds were evaluated by docking them into the binding site. Eight compounds were chosen to be synthesized and four to be purchased. The Suzuki-Miyaura cross-coupling reaction was used to couple the isoxazole core to different aromatic substituents, producing 3,5-disubstituted isoxazoles. The reactions mostly succeeded, but the yields were uniformly low. Developing the reaction using different solvents and reaction conditions did not produce clear results. Thirteen compounds were tested for activity, including an intermediate product of the synthesis. Two of the compounds showed increased inhibition activity compared to the hit compound, with approximated IC50 values of 10 and 40 μM, respectively. The knowledge gained from these studies can be used to further develop more efficient inhibitors.
  • Laustio, Netta (Helsingin yliopisto, 2018)
    During the past few decades, the explosion of discovery in cancer and immunological research has led to the increased understanding of the interactions between the immune system and tumors. These developments have provided vital information about the immune system’s role in cancer development. It is evidenced that the immunity system is capable to distinguish tumor cells from normal tissue by recognizing tumor antigens that are exclusively expressed on tumor cells or are presented in greater amounts on tumor cells than normal cells. Consequently, the immune cells start to attack tumors for protecting the host. The possibility to use the immune system as a weapon against cancer cells leaded to the promising innovation – cancer immunotherapy – which aims to activate the body’s own immune system and its components to mount antitumor immune responses for eliminating cancer cells. The antitumor efficacy and high safety profile of several immunotherapeutic strategies have already been demonstrated thereby resulting in their integration into clinical practice. However, most patients have not benefited from cancer immunotherapy as a single treatment. In this regard, new innovative methods are clearly needed to overcome the obstacles hindering the clinical success of this field. Therapeutic cancer vaccines are emerging as attractive immunotherapies currently being evaluated in both pre-clinical and clinical studies. The purpose of cancer vaccines is to eradicate tumor cells by eliciting antitumor CD8+ T cell responses against the injected tumor antigens. Due to the ability to specifically kill tumor cells and simultaneously trigger immune responses against tumor antigens via direct oncolysis and by encoding transferred tumor antigens, oncolytic viruses are of significant interest for being used as in situ cancer vaccines. Despite these unique properties, several factors such as tumor immunosuppression and immune tolerance to targeted tumor antigens resembling antigens of normal tissues hamper the use of oncolytic vaccines in clinic. Instead of focusing only on CD8+ T cells, it has been suggested that giving more attention to CD4+ T helper cells, which are required for priming and expansion of CD8+ T cell responses, could be the key to improve the efficacy of cancer vaccines. Researchers have also demonstrated that an ongoing antigen-specific CD4+ T cell response can lead to the bystander activation of surrounding T cells with unrelated antigen specificities. Based on this theory, the hypothesis of this study was to employ the pre-existing immunological CD4+ memory against infectious pathogens in generating bystander CD8+ immunity against solid tumors. In this study, mice transplanted with poorly immunogenic B16-OVA tumors were pre-immunized with the chosen vaccine to induce immunological CD4+ memory against an infectious pathogen. Tumors were then treated with already developed cancer vaccine, which was peptide-coated conditionally replicating adenovirus (PeptiCRAd) complex. PeptiCRAd was constructed by electrostatically coating adenovirus with both pathogen-derived and tumor-derived peptide. The intratumorally injected double-coated PeptiCRAd complex was assumed to activate peptide-specific T cells and thus, result in anti-pathogen CD4+ T cell recall responses and the following bystander activation of antitumor CD8+ T cells, which can then mount an effective immune response to destroy cancer cells. The efficacy of this treatment was observed in pre-immunized mice by measuring the growth of injected tumors. The experiment was repeated identically with non-immunized naïve mice to see the difference in the results. The immunological background of this treatment approach was investigated by analyzing mouse tissue samples with standard immunological techniques including ELISA, IFN-γ ELISPOT and flow cytometry. This study showed that long-term immunological memory against the pathogen was successfully accomplished and the strongest inhibition of tumor growth in pre-immunized mice was achieved with double-coated PeptiCRAd, whereas the antitumor efficacy was not seen in naïve mice. Additionally, a new ex vivo method to detect pathogen-specific CD4+ T cells from spleen was developed and the stimulation of cell-mediated immunity by this treatment was supported by finding the highest levels of pathogen-specific CD4+ Th1 cells from mice treated with double-coated PeptiCRAd. Some encouraging results concerning the beneficial immune cell composition of tumors and tumor draining lymph nodes were also obtained from other performed experiments. Though further immunological analyses are required for understanding the precise mechanisms of action behind the treatment, the increased immunogenicity and antitumor efficacy of double-coated PeptiCRAd can still be considered as a consequence of the bystander effect, which can possibly be utilized for developing improved strategies to win the fight against cancer.
  • Honkasalo, Oona (Helsingin yliopisto, 2018)
    Cancer immunotherapies aim to target the immune defence mechanisms of the body specifically and efficiently against the tumour tissue. Cancer vaccines and oncolytic viruses are forms of active immunotherapies, which require patients having a properly functioning immune system. The vaccines are based on the administration of tumour antigens into the body to which the immune system reacts. However, often the response is not robust enough. The oncolytic viruses in turn kill the cancer cells which causes the release of antigens from the tumour tissue. Viruses usually elicit a strong immune response but sometimes it is targeted too much against the virus instead of the tumour. Oncolytic vaccine is a composition of an oncolytic virus and a cancer vaccine. Tumour antigens can be coded to the genome of the virus therefore, when the virus invades tumour cells they start to produce the antigens. Eventually the cancer cells are also destroyed due to viral replication. The antigens can be tumour-associated that is, they are expressed in healthy tissues too. Their usage is not always efficient which is why an interest towards utilizing tumour-specific antigens has been increased. Considering the expression of antigens, tumour tissue is very heterogenous and distinctive between patients. Hence, utilizing mutated patient unique neoantigens would enable the development of personalized tumour-specific oncolytic vaccines. Genetic modification of viruses is complicated thus, an easier way to insert the neoantigens to the virus has been invented. The developed oncolytic vaccine platform is called PeptiENV, and it is designed to use with enveloped viruses. The idea is to fuse tumour-specific antigens onto the envelope of the virus and eliminate the need of gene insertion. The aim of this study is to investigate in vivo the efficacy of PeptiENV in preventing tumour growth and eliciting a tumour-specific immune response. An object is also to observe survival times of the treated animals. Furthermore, the preservation of infectivity is studied in vitro. The research was executed with two potential oncolytic viruses, vaccinia virus (VACV) and herpes simplex virus type 1 (HSV-1). The PeptiENV complex was formed by using an artificial tumour antigen, ovalbumin epitope SIINFEKL, which was attached to the viral envelope with cell penetrating peptide (CPP) or cholesterol anchor. The preservation of infectivity was examined by measuring cell viability of PeptiENV infected cells. Animal experiments instead were performed with a mouse melanoma model created with B16-OVA cells, which express ovalbumin and therefore the antigen epitope SIINFEKL. PeptiENV was compared to control treatments which were virus, SIINFEKL peptide and complexation medium only. Treatments were administered as intratumoural injections. Tumour growth was followed by measuring the size of implanted tumours every other day. With flow cytometry, tumour-specific immune response was assessed by acquiring the relative amount of SIINFEKL-specific CD8+ T cells in the tumour tissue. Euthanizing dates were registered in order to observe the survival of the mice. According to the in vitro results, conjugation of peptides to the virus does not affect infectivity. In addition, the in vivo studies show that PeptiENV VACV CPP prevents tumour growth the most. Difference in tumour growth between PeptiENV VACV CPP and control treatments is significant. Mice injected with the same treatment also lived considerably longer than mice injected with virus, peptide or medium only. Also, PeptiENV HSV-1 hinders tumour growth distinctly more than virus only and slightly more than SIINFEKL only, but unfortunately it did not have an evident impact on the survival time. In both experiments, the PeptiENV treatment elicits the largest proportional amount of SIINFEKL-specific CD8+ T cells. In other words, PeptiENV engenders a tumour-specific immune response. In the PeptiENV VACV study the difference to control treatments is clearer than in the PeptiENV HSV-1 study. At present, the PeptiENV platforms performs better with VACV than HSV-1. With further investigations however, the results can be verified and improved. All in all, the results are encouraging. The PeptiENV platform shows great promise for being a part of personalized cancer immunotherapy developments in the future.
  • Asikainen, Arsi (Helsingin yliopisto, 2018)
    Breast cancer is the most common cancer in women worldwide and the number of new events is on the increase. Like many other serious diseases, breast cancer reduces patient’s health related quality of life (HRQoL) and breast cancer treatment burdens our society. Examination of breast cancer patient’s HRQoL makes it possible to calculate how effective breast cancer treatments are. Nevertheless, only cost-effectiveness analysis would further help us allocate the resources of our society in the best way possible. The aim of this study was to produce research about breast cancer treatment’s effects on patient’s HRQoL and to compare generic 15D- and EQ-5D-5L-instruments. The results can be used in the future research and the study might be useful, when it’s time to develop international protocol for measuring HRQoL. The study population included 152 breast cancer patients who were treated in HUCH and whose HRQoL were measured by 15D-, EQ-5D-5L- and VAS-instruments. All measurements were done twice, first before the treatments and then six months after the beginning of the treatments. 89 (58.6 %) patients answered both 15D-questionnaires and 81 (53.3 %) patients answered to both EQ-5D-questionnaires. 57 (37.5 %) patients didn’t respond to any questionnaire. Only some background information was available of this population. The average HRQoL for breast cancer patients’ was 0.92 before the treatments and 0.88 six months after the beginning of the treatments when measured by 15D. The same average HRQoL was 0.86 before the treatments and 0.80 six months after the beginning of the treatments when measured by EQ-5D-5L. During six months’ period, patients HRQoL reduced (-0.04) when it was measured by 15D and (-0.06) when it was measured by EQ-5D. The changes of HRQoL were clinically important (The minimum important change, MIC > ± 0,015) when measured by 15D. HRQoL reduced more with patients who received a mastectomy than with patients, who received a breast conserving surgery according to both instruments. According to the results, the chosen instrument has an effect of breast cancer patients’ HRQoL. It means that the chosen instrument also has an effect of treatment’s effectiveness. 15D offers higher HRQoL values, but EQ-5D offers a greater change in patient’s HRQoL. HRQoL was measured by two different generic instruments in two different times, which was assumed to be the strength of this study. The new 5L-version of EQ-5D-instrument was also used. This is possibly the first time, when 5L is used in this type of study.
  • Sjöberg, Madeleine (Helsingin yliopisto, 2018)
    Cancer afflicts an ever-growing number of people globally each year. In part due to a complex pathophysiology where much is still unknown, the need for new cancer treatments has been persistent, fuelled further by the issue of treatment resistance. An emerging field holding much promise in oncology is immunotherapy, a subgroup of which is oncolytic virus treatments. These treatments utilize the inherent or acquired ability of certain viruses to selectively replicate in tumor cells to fight cancer. One of these viruses is the adenovirus. With these viruses it is possible to modulate the immune response e.g. through the expression of certain genes. The thesis focuses on genetically arming an oncolytic adenovirus in an effort to enhance treatment efficacy. The transgene of choice is the CD40 ligand (CD40L), a costimulatory molecule capable of aiding in the development of systemic antitumor immunity. Adenoviruses have previously been designed expressing the CD40L, however, a novel aspect was introduced with the design and incorporation of a soluble a form of the protein. The main aim of the study was the construction of four functional oncolytic adenoviruses, encoded with either the human or mouse variants of the two CD40L proteins (full-length and soluble). Successful completion required protocols for the cloning, bacterial colony screening, and primary virus production to be established. Insertion of the CD40L transgenes into the E3-gp19k region of the chosen Ad5Δ24 backbone was first attempted with the traditional approach of homologous recombination. The method that ultimately proved successful was a one-step Gibson Assembly® reaction. Screening the bacterial colonies with colony polymerase chain reaction, the potential CD40L positive clones underwent restriction analysis to affirm the presence of the transgene in the viral genome, as well as the retainment of critical elements. Two out of three recombined plasmids carrying the full-length CD40L proceeded to transfection and virus propagation in A549 cells, after which the presence of the adenovirus and CD40L expression was confirmed with immunostaining. Finally, a protocol was successfully established by the construction of one of the intended four viruses. The protocol entails all the main steps from cloning until primary virus production, additionally offering the option of applying it to the genetic arming of the Ad5Δ24 with other transgenes of interest. In terms of future perspectives for the project, following construction of the remaining viruses, the intentions are to validate transgene expression and functionality for all constructs, as well as compare the immunogenicity between the full-length and soluble CD40L. In the event of promising results, the project will hopefully proceed to in vivo studies.

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