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Ambient mass spectrometry includes methods where ions are produced outside of the mass spectrometry in atmospheric pressure direct from the surface of the sample without sample preparation. The first and most popular ambient ionization methods are DESI, desorption electrospray ionization and DART, direct analysis in real time. DAPPI, desorption atmospheric pressure photoionization is an ionization method where samples are desorbed with hot vapor from surface and then ionized by photoionization. The aim of this study was to develop desorption atmospheric pressure photoionization method in transmission geometry. In transmission geometry hot vapor for microchip is directed through metal or polymer meshes to mass spectrometer inlet. Liquid samples can be analyzed either by soaking the mesh to liquid sample or apply a sample droplet to the mesh. Hot vapor desorbs analytes from the mesh and analytes are ionized in a gas phase by photoionization using VUV lamp. In this method optimal positioning of the mesh and the microchip was determined. Additionally optimal microchip heating power, dopant flow rate, nebulizer gas flow rate, capillary voltage and drying gas parameters were determined. Optimized method was applied for analyzing standard samples, vitamin juice samples and milk samples. According the analysis with authentic samples, transmission mode DAPPI can be applied for analyzing liquid samples without sample preparation. According the analysis with standard samples, transmission mode DAPPI can be applied for extraction of hydrophobic analytes from water samples. Comparing to conventional DAPPI, in transmission mode DAPPI spectra, intensities of the background ions are lower resulting higher signal-to-noise ratios with transmission mode DAPPI.

Subanesthetic-dose ketamine, an N-methyl-D-aspartate receptor (NMDAR) blocker, exerts rapid antidepressant effects that sustain long after its elimination from the body. The precise mechanism remains unknown, but regulation of TrkB (tropomyosin receptor kinase B), ERK (extracellular-regulated kinase 1 and 2), GSK3β (glycogen synthase kinase 3β) and mTOR (mammalian target of rapamycin) signaling within the prefrontal cortex (PFC) have been deemed important for its antidepressant-like effects in rodents. In addition, activation of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) is thought to be an important step in its mechanism. Nitrous oxide (N2O), another NMDAR antagonist and a putative rapid-acting antidepressant, regulates the same molecular pathways as ketamine in the rodent PFC. The fast pharmacokinetics of N2O have been exploited to show that markers of neuronal excitation, including phosphorylation of ERK, are upregulated in the PFC during its acute pharmacological effects (NMDAR blockade), while regulation of TrkB, GSK3β and P70S6K emerges only upon N2O withdrawal. In the first part of this study, we investigated the N2O-induced biochemical changes associated with neuronal excitation and BDNF-TrkB signaling in the PFC and further, the requirement for AMPAR activation in inducing them. We focused on the effects seen after the acute pharmacological effects of N2O. N2O (65% for 20 min) was administered to adult male C57BL/6 mice with or without pretreatment with AMPAR antagonist (NBQX, 10 mg/kg) and PFC samples were collected 15 minutes after stopping N2O delivery. Within this time N2O is expected to be completely eliminated. The brain samples were analyzed using western blot, enzyme-linked immunosorbent assay and quantitative reverse transcription PCR. We observed that N2O increased levels of phosphorylated TrkB, GSK3β and P70S6K, and these effects were not attenuated by NBQX pretreatment. At the same time, we observed a decrease in the levels of phosphorylated ERK, which was attenuated in mice that received NBQX prior to N2O. Tissue levels of BDNF protein or messenger RNA (exon IV) were not different between control and experimental groups. These results indicate that the mechanism of N2O is associated with TrkB and ERK signaling that are regulated independently of each other. It appears that AMPAR activation is not required for TrkB signaling, although it might play a role in ERK signaling. Further, N2O-induced TrkB phosphorylation in the PFC is not associated with changes in total levels of BDNF. In the second part of the study, we aimed to search for new ketamine-like NMDAR blockers with antidepressant potential. Ketamine was used as a query compound for in silico substructure search to find commercial ketamine analogs. The retrieved ketamine analogs were filtered by their computed ADMET properties and then further screened virtually by docking them to the pore region of NMDAR complex (protein data bank code: 4TLM), around the predicted binding site of ketamine. Finally, we sought to study if selected ketamine analogs could elicit ketamine-like effects on TrkB and ERK signaling in mouse primary cortical neurons. However, we did not proceed to test the analogs since ketamine (positive control) did not show any effects on TrkB or ERK phosphorylation in our culture. Overall, this study advances the understanding of the mechanism of N2O, possibly giving new insight of the antidepressant mechanisms of NMDAR-blocking agents more generally. Additionally, we found promising ketamine analogs that await experimental testing.

Rodent studies indicate that the effects of pharmacological antidepressant treatments depend on the TrkB (tropomyosin-related kinase B) receptor of the neurotrophic factor BDNF (brain-derived neurotrophic factor). However, the mechanism by which TrkB signaling becomes active remains disputed. Our group proposes that the activation of TrkB signaling is a result of an indirect physiological adaptation to the drug treatment, which is supported by observations made with rapid-acting antidepressants ketamine and nitrous oxide. Previous studies indicate that the immediate effects of the drugs are followed by a sedative state resembling deep sleep, during which TrkB signaling becomes active. The sedative state is accompanied with a drop in core body temperature, and preliminary findings indicate that preventing the drug-induced hypothermia blocks TrkB signaling in the cortex.    The purpose of this study was to investigate the effect of ambient temperature on TrkB signaling in the hippocampus following nitrous oxide administration. Nitrous oxide (65 % ad 100 % O2) was administered to adult male mice for 20 minutes. After the drug treatment the animals were kept in different recovery conditions: room temperature or a heightened ambient temperature of approximately 36 °C for 15 minutes. Following the recovery, the animals were euthanised, and hippocampus samples were collected from the animals. Levels of BDNF and TrkB signaling were analysed with ELISA and western blot, respectively.    Nitrous oxide caused a significant drop in core body temperature, but this was not accompanied with increased BDNF levels or TrkB signaling. Evidence suggests that hippocampal atrophy contributes to depression, but the acute effects of antidepressant treatments on TrkB signaling in this brain area appear to be less pronounced than those seen in the prefrontal cortex. The findings indicate that nitrous oxide has a replicable hypothermic effect, but this is not associated with increased TrkB signaling in the hippocampus.

For their good clinical value and supply the use of biological medicines has increased in the treatment of inflammatory bowel disease (IBD). However, biological medicines are often more expensive compared to traditional small molecule medicines. More inexpensive biosimilars, shown to be clinically equal to the corresponding biological original products, can be used to reduce medication costs as part of rational pharmacotherapy. Although patients’ perceptions about biosimilars may affect the treatment adherence and outcomes of the use of these medicines, only a few studies have been published on this area. The primary aim of the study was to study IBD patients’ perceptions of the features of biosimilars and biosimilars’ suitability for their own treatment. The secondary aim was to study biological medicine users’ perceptions of the suitability of biosimilar switch by a physician for their own treatment. In addition, sources of medicines information of the users of biological medicines were studied. The data of the study were based on a cross-sectional survey conducted by the University Pharmacy and University of Helsinki in January 2021. Research Newsletter and response link were delivered per an email to University Pharmacy’s loyal customers and electronically communicated via the Association of Rheumatism and the Association of IBD and other intestinal diseases. The study comprised of the responses of adult outpatients with IBD (n=979) who were using original biological medicines (n=120), biosimilars (n=30) or traditional small molecule medicines (n=829) (medicine user groups). The sum variables constructed by factor analysis based on the study aims were used as outcome variables. Differences between medicine user groups and the effects of other background variables were analyzed by bivariate and multivariate analysis using SPSS Statistics software. Most of the patients (70–97 % depending on the medicine user group) trusted biosimilars’ features to be equal to the corresponding original biological medicines. However, more than half of the patients (53–67%) did not know whether they would like to physician to prescribe biosimilar to them rather than the original biological medicine. Of the users of original biological medicines, 71 % did not want to be switched to a biosimilar if the current medication was working well. Biosimilar users had more positive perceptions about biosimilars and switching compared to other medicine user groups. Physician’s perception played an important role in biologic switching. Several factors that may affect perceptions about the features of biosimilars and their use in patient’s own treatment were identified. These included, in particular, previous user experience with biosimilars and having information about them. Overall, patients had positive perceptions about the features of biosimilars but had uncertainties about the use of biosimilars, especially, for the treatment of their own disease and when switching medication. More research is needed on the perceptions among different patient groups, and on the need and optimal form of medication information on biological medicines, biosimilars and their switching.

The health and social services reform will enter into force in its entirety from the beginning of 2023. With the reform, the responsibility for organizing social and health care will be transferred from municipalities to the responsibility of 21 wellbeing services counties. At the time of writing this thesis, the changes to the medical legislation brought by the reform have not yet been published. Hospital pharmacies and dispensaries take care of Finland's public pharmaceutical services. The tasks of public pharmaceutical services include pharmaceutical logistics tasks, non-industrial pharmaceutical manufacturing, and the preparing of medicines, as well as pharmaceutical expert tasks and services.The aim of this study is to find out the opinions of current hospital district managers, medical directors, and hospital pharmacists about how future pharmaceutical services should be organized in the upcoming wellbeing service counties. The study was conducted using an electronic structured questionnaire, which was sent in October 2021 by e-mail to the heads of all Finnish hospital districts, medical directors, and hospital pharmacists. The survey also included Åland and the Joint Municipal Authority for Social and Healthcare in Central Uusimaa (Keusote). The questionnaire consisted mainly of Likert-scale questions, but the questionnaire also had open answer fields to which respondents were able to add comments and refine their answers. The questionnaire was evaluated by several experts and piloted by two experts. The questionnaire consisted of seven different sections, which addressed the number and concentration of hospital pharmacies and dispensaries, clinical pharmacy services and medication safety, pharmaceutical purchasing and formulary, automation and information systems, co-operation in wellbeing service counties, and pharmaceutical services in a state of emergency and limited resources. The overall response rate to the survey was 50% (n = 34/68). 79 per cent (n = 19/24) of hospital pharmacists and 35 per cent (n = 15/43) of managers and medical directors responded to the survey. Responses were received from all hospital districts, Åland and Keusote. Based on the responses, it is hoped that the activities of hospital pharmacies will be mainly concentrated in wellbeing service counties so that the services would not move too far. Co-operation in individual pharmaceutical service activities could take place in collaborative areas or nationwide. The current number of hospital pharmacies was thought to be sufficient, but the operation of individual dispensaries could be closed or transferred to the administration and coordination of a hospital pharmacy in the area. It is hoped that clinical pharmacy services will be increased, and medication safety officers are desired for at least all wellbeing service counties. It is hoped that purchasing will be centralized nationwide, especially for expensive and rare pharmaceuticals, but the procurement of pharmaceutical formularies could be done by collaborative area, and the formation of the formularies could be done by wellbeing service county or collaborative area. It is hoped that automation and technology will increase in pharmaceutical services and that information systems will become more integrated. Increasing co-operation both between hospital pharmacies and within wellbeing service counties, for example between community pharmacies, was advocated. In the future, resources should be focused on pharmaceutical services personnel and their training, as well as on automation and technological solutions. It is hoped that the crisis preparedness of the pharmaceutical services will be increased in the future.

Annually thousands of Finnish children are placed in foster care as a measure of child welfare. Institutional foster care is provided by child welfare institutions. Social and health care professionals, such as bachelors in social services, youth workers, practical nurses, and registered nurses working in these institutions are responsible for carrying out the children’s medication treatment. Depending on completed basic and additional training and work experience, the personnel have varying competencies in medication treatment. The aim of this study was to provide research-based information on the challenges and development needs related to the safe and rational medication treatment of children in foster care living in child welfare institutions. This was studied from the point of view of institution personnel and social workers responsible for placement decisions. Furthermore, the study investigated the medication use process of children in institutional foster care and, how the child’s health and medication treatment related needs are considered in the placement process and in selecting the foster care place. The study was conducted as a qualitative study using focus group discussions (FGDs) carried out to child welfare social workers (n=1) and child welfare institution personnel (n=10) during November and December 2022. Semi-structured focus group discussions (n=3) were conducted over the video conferencing software Microsoft Teams®. The participants (n=11) were recruited through child welfare services of Central Uusimaa Wellbeing County. Qualitative content analysis was used to analyze the data. According to the focus groups, medication treatment is common among children living in child welfare institutions. In the daily life of child welfare institutions, there were challenges and development needs related to medication treatment and medication use process. The main challenges and development needs were the lack of up-to-date information regarding the children’s health status and medication, challenges in organizing acute medication treatment, the personnel’s up-to-date drug administration permissions and varying competencies in medication treatment. In addition, fragmented overall responsibility of the medication treatments and poor access to healthcare presented challenges for organizing and carrying out medication treatment for the children. Medication use process during foster care appeared to be fragmented and it was seen to be susceptible to errors. According to this study, the possibilities for taking children’s health status and medical needs into account at the point of placement were scarce. These needs often did not guide placement decisions. Considering the medication use process and medication safety, it is essential that child welfare institutions have access to up-to-date health and medication information of the children. To achieve the best interests of the children, organizing acute medical treatment should be facilitated and the overall responsibility health care and medication therapy clarified. In addition, seamless access to the necessary healthcare must be ensured for children in foster care. Considering medication safety, it is important to ensure that the personnel of child welfare institutions have up-to-date competencies in medication treatment that meet the needs of the children.

The aim of this Master's Thesis was to assess experiences of access to medications and follow up services. The aspects studied were: access to medications from the public's perspective, also in relation to availability of follow-up services and support for self-management in long-term medications, and difficulty to buy necessary medicines due to economic reasons. The respondents were also asked to identify needs for developing new customer-oriented services for follow up of treatments. The data were collected during December 2013 and January 2014 by an email survey to those registered in the loyal customer program of University Pharmacy. The data were analyzed by using the statistical programme SPSS. Responses to open-ended questions were analyzed (a preliminary analysis). Respondents' age, gender, area of residency and financial situation were used as background variables. 606 responses were received (84% women, 16% men). The mean age of the respondents was 53.5 years and 91% had at least one disease or symptom diagnosed by a doctor. Almost all (93%) used some medicine or vitamin product. Eleven percent of the respondents reported that they had not been able to purchase a medicine they needed due to poor personal financial situation. A majority (85%) of the respondents perceived their health status as good. The average number of visits at the doctor during a one year period was 5.5. About 22 % of the respondents reported that they were not able to get an appointment when they needed it. About half of the respondents had regular health controls by a doctor. The respondents indicated a wish that getting the appointment regularly should be easier and that there should be time to have a holistic discussion on one's care. About half had a personal doctor and 42% had a medication card. The most common ways to self-monitor one's care were by observing general health status, measuring blood pressure and weight. Almost two-thirds (63%) discussed the monitoring results with their doctor. Many respondents reported in the open comments that they did not have instructions for self-monitoring and there was no healthcare provider to share the results with. The respondents wanted have more information concerning the reasons to use medicines, and the benefits of a long-term medicine use. They also wanted to know more about adverse effects and interactions, as well as about non-pharmacological treatment options. The prescriptions were most commonly renewed at the doctor's office (47%).The respondents also expressed a wish to have more options to contact their healthcare providers, e.g., though electronic services (online doctor, email counselling). There are limitations in the Finnish health care system from the medication management's perspective. Aspects needing improvement include access to regular controls and follow-up services, having more options to contact healthcare providers, also through electronic services, having better access to information on diseases and medication, and finally, improve caring for people's health concerns in a holistic way.

Parkinson's disease is a progressive degenerative brain disease that causes degeneration of dopaminergic neurons in the substantia nigra. Characteristic motor symptoms in Parkinson's disease are caused by dopamine deficiency in striatum. Tyrosine hydroxylase (TH) is the enzyme that catalyzes the rate-limiting step in the dopamine biosynthesis. Because of this TH has a significant role in the function of the dopaminergic system. TH activity is regulated by several mechanisms. The most important regulatory mechanism is phosphorylation of TH protein by spesific protein kinases. Alterations in the function of TH have been associated with Parkinson's disease. The most prominent findings are decreased TH protein and TH mRNA content in the nigrostriatal dopaminergic neurons. A possible pathogenic role of TH in Parkinson's disease has also been suggested. In addition TH might be a potential therapeutic protein for gene therapy. One possible approach is viral vector-mediated gene transfer of TH gene directly into the brain. Simultaneous gene transfer of TH gene and neurotrophic factor gene could both enhance dopamine synthesis and prevent remaining dopaminergic neurons from dying. None of the current treatments of Parkinson's disease can halt or retard dopaminergic neuron degeneration. Novel treatments are being developed and amongst other strategies neurotrophic factors have proven promising candidates for the treatment of Parkinson's disease. Member of CDNF/MANF family of neurotrophic factors, cerebral dopamine neurotrophic factor (CDNF), is currently being studied. Previous studies have demonstrated the neuroprotective and neurorestorative effects of CDNF but more research is needed for optimal administration technique and dose. The aim of this work was to study the neuroprotective effect of AAV vector-mediated delivery of CDNF (AAV-CDNF) in a rat model of Parkinson's disease. Rats' brains were unilaterally lesioned with intrastriatal injection of 6-OHDA two weeks after viral vector injections and amphetamine-induced rotational behavior was monitored for ten weeks. The CDNF protein expression after intrastriatal AAV vector-mediated gene transfer was analyzed with immunohistochemical staining of brain sections. We confirmed that CDNF protein is expressed in rat brain after intrastriatal injection of AAV-CDNF. AAV-CDNF treatment also reduced the amphetamineinduced ipsilateral rotations nearly as much as AAV-GDNF treatment. AAV-CDNF treatment also had an effect on the amount of remaining TH-immunoreactive cells in the substantia nigra pars compacta and the optical density of striatal TH-immunoreactive fibers but these results did not reach statistical significance. The immunohistochemical measures did not correlate completely with the behavioral data and further studies are needed to confirm the results obtained here. The results of this research support the conclusion that AAV-CDNF treatment has a neuroprotective effect on nigrostriatal dopaminergic neurons.

Oxygen has been used as a medicine since the 18th century and is widely used as prescription and over-the-counter (OTC) medicine. Especially with global COVID-19 pandemic, which started in 2020, the demand for medicinal oxygen has increased significantly and the quality of medicinal oxygen has become increasingly important. Only few studies have been published on the critical process and quality parameters of gases and their impact on product quality. Because oxygen is classified as a medical product, it must be manufactured in accordance with good manufacturing practices regulations (GMP). One part of EU and other GMP guidelines is mandatory annual product quality review (PQR) which must assess the critical quality and process parameters as well as their trends. The aim of the study was to define the critical process and quality parameters for the medicinal oxygen filling process and to analyze the process control, stability, and capability for annual PQR using process data. Process stability and the state of control of processes were assessed using statistical quality and process management tools, such as the Shewhart control diagram and process capability index. Studied process parameters included vacuum level and pressure measured by the filling equipment. Evaluated quality parameters included analysis pressure, O2 and H2O contents. The results of the study showed that the process is not stable and there was a lot of variation between the parameters. Most variation was detected between different cylinder volumes and filling and analysis ramps in all parameters and between different weekdays in H2O content. However, all parameters remained within the specification limits and the Cpk values of all critical parameters were good. By analyzing the data, many variables that can affect the parameters and add variation to the process data can be identified. Based on the results, the necessary measures to improve and optimize the process and quality was identified. In order to stabilize processes and improve performance, the demonstrable variation in process data should be reduced, for example by harmonizing operating methods. According to the study, it was also possible to assess the revalidations required for the process.

Plasmidit ovat geneettisiä elementtejä, joita voidaan käyttää esimerkiksi geeninsiirtovektoreina. Transposonit ovat DNA-fragmentteja, joilla on kyky siirtyä genomissa paikasta toiseen. Tutkimuksessa käytettävä transposoni on piggyBac, joka on eristetty tupsumetalliyökkösen (Trichoplusia ni) soluista. Transpositiossa piggyBac tunnistaa ITR-osat (käännetty terminaalinen toistojakso) siirtäen osien välissä olevan DNA:n. Tutkimuksen tavoitteena oli tuottaa kaksi erilaista plasmidia. pAc5.1-piggyBac-plasmidiin sisällytettiin piggyBac ja pMT-In-EGFP-PB-ITR-plasmidiin ITR-osat sekä niiden väliin hygromysiiniresistenssigeeni sekä EGFP-geeni. BTI-Tn-5B1-4-solujen DNA:sta eristettiin piggyBac sekä ITR-osat ja ne siirrettiin plasmidiin pTOPO-piggyBac-R. Tästä plasmidista irrotettiin erilleen piggyBac ja ITR-osat, joista välivaiheiden kautta rakennettiin lopulliset plasmidit. Plasmidit rakennettiin pitkälti pilkkomalla DNA-fragmentteja restriktioentsyymeillä ja yhdistämällä niitä ligaatiolla. Plasmideja tuotettiin suurempia määriä siirtämällä niitä transformaation avulla E.Coli-soluihin lämpöshokkimenetelmällä ja eristämällä tämän jälkeen saadut plasmidit. Tuotettujen plasmidien onnistuminen varmistettiin pilkkomalla ne restriktioentsyymeillä ja tutkimalla DNA-fragmenttien kokoa agaroosigeelielektroforeesilla. Plasmidinäytteet myös sekvensoitiin osittain. Banaanikärpäsen (Drosophila melanogaster) S2-solut transfektoitiin kehitetyillä plasmideilla ja solukonsentraatioita sekä elinkelpoisuutta mitattiin 8 päivän ajan transfektion jälkeen. Tavoitteena oli hyödyntää EGFP-geeniä fluoresenssimittauksiin. Solunäytteisiin lisättiin kokeen aikana hygromysiini, jotta voitiin selvittää, olivatko viljellyt solut saavuttaneet hygromysiiniresistenssin. Tutkimuksen tuloksena plasmidit saatiin kehitettyä, mutta solukokeiden tulokset jäivät epäselviksi. Solunäytteissä ilmeni kasvatuksen aikana kontaminaatioita. Lisäksi EGFP-osia ei voitu luotettavasti mitata käytössä olleella laitteella. Transfektio tulee siis toistaa transposonisysteemin toiminnan tutkimiseksi. Lisäkokeilla voidaan selvittää tarkemmin kehitetyn transposonisysteemin mahdollisuuksia sekä toiminnan yksityiskohtia.