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  • Jokinen, Birgitta (2010)
    Angiogenesis may be regarded as one of the most important phenomena involved in basic physiology as well as in numerous pathological conditions. Angiogenesis is a multistep process involving the balance of pro- and con-angiogenic factors. Several studies have suggested that angiogenesis is regulated in vitro and in vivo by peptides thymosin ȕ4 (Tȕ4) and tetrapeptide Ac-SDKP (N-acetyl-seryl-aspartyl-lysyl-proline). There are also studies supporting the view that Ac-SDKP, a peptide fragment is released from the proline-containing C-terminus of Tȕ4 (43-mer) by hydrolyzing prolyl oligopeptidase (POP). POP is a widely existing serine protease cleaving oligopeptides of no longer than 30 amino acids. Thus, Tȕ4 should first be cleaved into a shorter peptide by some other, yet unknown peptidase. POP has been mostly studied in memory and learning disorders as well as in neurodegenerative diseases. The true physiological character of POP is still unresolved. In this Master's thesis, the associations of the factors involved in angiogenesis are reviewed in the literature part as well as the character, presence and function of the angiogenic molecules 7ȕ4, Ac-SDKP and POP. In the experimental part attempts were made to find whether POP and Tȕ4 increase Ac-SDKP formation and capillary tube network and consequently, whether the POP activity, tetrapeptide and capillary formation could be inhibited by the proline-spesific POP inhibitor KYP-2047. The study had two phases. The first phase included POP activity and Ac-SDKP measurements(time period 0-180 min) with Wistar rat kidney homogenates. Study groups were 0,1 and 0,5 µM KYP-2047 (+2 µM Tȕ4), 1:20 (0.625 µM) human recombinant POP (+ 2 µM Tȕ4), 2 µM 7ȕ4 (pos. control) and raw homogenate (neg. control). The second phase involved the study of capillary formation (time period 0-180 min) with primary endothelial HUVECs on a 48-well plate seeded with 50 000 cells/well on an extracellular membrane mimicking MatrigelTM Matrix dissolved in DMEM. Study groups treated with fetal bovine serum and antibiotics were 5 and 10 µM KYP-2047 (+4 µM Tȕ4), 1:20 (0.625 µM) human recombinant POP (+4 µM Tȕ4)4 µM Tȕ4 (pos. control) and DMEM (neg. control). The wells were cultured and capillary formation photographed with a light microscope using a digital camera. All experiments were repeated four times, and each study group in wells was measured in triplicate. Enclosed capillaries were counted manually and statistical tests were performed. 7ȕ4 along with POP participated in the formation of AC-SDKP in the kidney homogenates. Cultures of primary endothelial cells on Matrigel resulted in clear capillary formation in Tȕ4 and POP groups. KYP-2047 had a strong POP-inhibitory activity on antiangiogenesis throughout the study resulting. Obviously, underlying mechanisms of angiogenesis and the function of the interaction between POP, Tȕ4 and Ac-SDKP in capillary formation require further studies.
  • Neittaanmäki, Eerika (2015)
    Actinic keratoses are premalignant skin lesions caused by sun UV-radiation. A small portion of these lesions progress into invasive squamous cell carcinoma over the years. Actinic keratoses are a growing problem in the healthcare around the world. P53 mutations are found in actinic keratosis and adjacent areas. Treatment options include surgical removal, cryotherapy, local treatment creams such as immunomodulators, and photodynamic therapy (PDT). Aminolevulenic acid (ALA) is an endogenous light-sensitizer used in PDT and methylaminolevulinate (MAL) and hexylaminolevulinate (HAL) are its esters. Either artificial light or daylight can be used as a light source in PDT. In PDT light activates the photosensitizer, which initiates a photochemical reaction and target cell destruction. The most common side effects of PDT are erythema and pain during treatment. The benefits of PDT are good cosmetic outcome and the possibility to treat large areas. In the present study 3 light-sensitizers (BF-200 ALA, MAL, HAL 0,2 % and 2 %) were tested on healthy volunteers to compare the skin irritation, pain and fluorescence caused by the treatment. Fluorescence intensity reflects PpIX production capacity. The second part of the present study was a clinical study comparing BF-200 ALA and MAL in treatment of actinic keratoses with daylight-PDT. Each patient received both light-sensitizers on opposite sides of the head and the results were evaluated 3 months after treatment. On healthy skin BF-200 ALA, MAL and HAL 2 % caused more irritation compared to HAL 0,2 %. HAL 2 % didn't differ from ALA and MAL-groups in severity of reactions, erythema or fluorescence photobleaching. In HAL-treated areas pain was smaller than in ALA- and MAL-groups. However, in lesional skin there might be differences in absorption, distribution and PpIX formation. Both BF-200 ALA and MAL were effective in daylight-PDT and there were no significant differences between the groups in either efficacy or pain caused by the treatment. Long-term follow up is still required to confirm if the results sustain.
  • Hannula, Mirva (2010)
    Prolyl oligopeptidase (POP, prolyl endopeptidase, EC 3.4.21.26) is a serine-type peptidase (family S9 of clan SC) hydrolyzing peptides shorter than 30 amino acids. POP has been found in various mammalian and bacterial sources and it is widely distributed throughout different organisms. In human and rat, POP enzyme activity has been detected in most tissues, with the highest activity found mostly in the brain. POP has gained scientific interest as being involved in the hydrolyzis of many bioactive peptides connected with learning and memory functions, and also with neurodegenerative disorders. In drug or lesion induced amnesia models and in aged rodents, POP inhibitors have been able to revert memory loss. POP may have a fuction in IP3 signaling and it may be a possible target of mood stabilizing substances. POP may also have a role in protein trafficking, sorting and secretion. The role of POP during ontogeny has not yet been resolved. POP enzyme activity and expression have shown fluctuation during development. Specially high enzyme activities have been measured in the brain during early development. Reduced neuronal proliferation and differentation in presence of POP inhibitor have been reported. Nuclear POP has been observed in proliferating peripheral tissues and in cell cultures at the early stage of development. Also, POP coding mRNA is abundantly expressed during brain ontogeny and the highest levels of expression are associated with proliferative germinal matrices. This observation indicates a special role for POP in the regulation of neurogenesis during development. For the experimental part, the study was undertaken to investigate the expression and distribution of POP protein and enzymatic activity of POP in developing rat brain (from embryonic day 14 to post natal day 7) using immunohistochemistry, POP enzyme activity measurements and western blot-analysis. The aim was also to find in vivo confirmation of the nuclear colocalization of POP during early brain ontogeny. For immunohistochemistry, cryosections from the brains of the fetuses/rats were made and stained using specific antibody for POP and fluorescent markers for POP and nuclei. The enzyme activity assay was based on the fluorescence of 7- amino-4-methylcoumarin (AMC) generated from the fluorogenic substrate succinyl-glycyl-prolyl-7-amino-4-methylcoumarin (Suc-Gly-Pro-AMC) by POP. The amounts of POP protein and the specifity of POP antibody in rat embryos was confirmed by western blot analysis. We observed that enzymatic activity of POP is highest at embryonic day 18 while the protein amounts reach their peak at birth. POP was widely present throughout the developmental stages from embryonic day 14 to parturition day, although the POP-immunoreactivity varied abundantly. At embryonic days 14 and 18 notably amounts of POP was distributed at proliferative germinal zones. Furthermore, POP was located in the nucleus early in the development but is transferred to cytosol before birth. At P0 and P7 the POP-immunoreactivity was also widely observed, but the amount of POP was notably reduced at P7. POP was present in cytosol and in intercellular space, but no nuclear POP was observed. These findings support the idea of POP being involved in specific brain functions, such as neuronal proliferation and differentation. Our results in vivo confirm the previous cell culture results supporting the role of POP in neurogenesis. Moreover, an inconsistency of POP protein amounts and enzymatic activity late in the development suggests a strong regulation of POP activity and a possible non-hydrolytic role at that stage.
  • Puranen, Pinja (2023)
    Medical devices play a crucial role in healthcare, yet their importance is underscored by the potential for adverse events that can lead to serious consequences for patients and users. As a result, manufacturers of medical devices are required to actively monitor the safety and performance of their devices after placing them on the market. In response to incidents involving medical devices and their safety, the European Union Medical Device Regulation (MDR) came into force in May 2021, bringing more emphasis to the post-market surveillance (PMS) of medical devices. In this study, the current state of the post-market surveillance system of medical device manufacturers in Finland was investigated by conducting an online questionnaire in 2023. A total of 30 medical device companies participated in the questionnaire for a return rate of 17%. The dataset included manufacturers of different sizes, producing medical devices of all risk classes. To identify differences in the use of post-market surveillance data sources between different types of companies, the two-sided non-parametric Kruskal-Wallis-Test was used. The rest of the data was analysed using descriptive analysis. The post-market surveillance data sources with the highest reported intensity of use included customer complaints and feedback, production monitoring, and regulatory intelligence monitoring, while Post Market Clinical Follow-up and health services research were used significantly less. Significant differences between manufacturers of different device risk classes were identified for three data sources; manufacturers of higher risk class devices were found to utilize these data sources to a higher extent than manufacturers of low-risk devices. The manufacturers of medical devices seem to utilise reactive post-market surveillance data consistently to a high extent. On the other hand, the results suggest that proactive post-market surveillance methods remain underutilised despite the introduction of the MDR. Medical device manufacturers also use post-market surveillance data sources to different extents, in particular with respect to the medical device risk class. Overall, the results indicate that the MDR is bringing more emphasis on post-market surveillance, which in turn has increased the workload of medical device manufacturers.
  • Pylkkänen, Sarita (2013)
    Harmful drug effects are common among older medicine users. Potentially harmful drugs for older people have been defined by different criteria. Potentially inappropriate medications (PIMs) defined by Beers criteria have been associated with adverse effects, increased costs, need of hospital care and disabilities. Drugs with anticholinergic properties (DAPs) are associated with anticholinergic side effects, cognitive decline and delirium. Psychotropic drugs have been associated with increased risk of falls and mortality. Concurrent use (≥3) of psychotropic drugs has been considered harmful for older people by the Swedish National Board of Health and Welfare. The aim of this study was to examine the prevalence of potentially harmful drug (PHD) use (DAPs, PIMs, or concurrent use of ≥3 psychotropic drugs) and the accumulation of PHDs in aged people living in nursing homes and assisted living facilities. The objective was also to investigate which patient characteristics are associated with PHD use or accumulation of these drugs. The cross-sectional data was collected in 2011-2012 as a part of a larger study "Reducing inappropriate, anticholinergic and psychotropic drug use among older residents in institutional care". The study population (N=326) consisted of ≥ 65-aged residents living in nursing homes or assisted living facilities in Helsinki (n=227) and Kouvola (n=99). The mean age was 83.5 years, 70.0 % were women and mean Charlson comorbidity index was 2.6. Residents were divided into four groups: aged using 1) DAPs, 2) PIMs defined by Beers criteria, 3) concurrent use of ≥3 psychotropic drugs and 4) no PHDs in use. Both those fulfilling any of these criteria and those fulfilling all the three criteria (accumulation of PHD) are described. Users in these groups were compared to the non-users. A majority of the residents, 78,8 % (95 % CI: 74,4 - 83,3) used ≥1 PHDs: 67,8 % (95 % CI: 62,7- 72,9) used ≥1 DAPs, 32,2 % (95 % CI: 27,1- 37,3) ≥1 PIMs and 32,2 % (95 % CI: 27,1-37,3) used ≥3 psychotropics concurrently. Of the residents, 41 (12.6 %) had a medication treatment that fulfilled all the three criteria of PHD use. These residents used significantly more PHDs than others (average mean 4,8 [range 3-7] vs. 1,6 [range 0-6]). The residents having the PHD accumulation were more often males and used more drugs than others. There were no statistical differences among the other characteristics of these groups. The most common PHDs were mirtazapine (n=66), lorazepam (n=64), oxazepam (n=62), ketiapine (n=58) and stimulant laxatives without opioids (n=58). Use of DAPs was associated with multimorbidity, use of PIMs with weaker health related quality of life, and use of psychotropics with younger age. High number of drugs was associated with all these criteria. Use and accumulation of potentially harmful drugs is common among the aged living in nursing homes and assisted living facilities. New means are needed to optimize drug treatments and to educate professionals taking care of these patients. Special attention should be paid on the use of antipsychotics, benzodiazepines, mirtazapine and stimulant laxatives.
  • Lindholm, Anni (2023)
    Biologisten lääkkeiden käyttö on merkittävästi lisääntynyt 2000-luvulla, mikä on hoidollisten hyötyjen ohella lisännyt lääkekustannuksia. Vaihtokelpoisten ja halvempien biosimilaarien käyttöä on edistetty koulutuksella, lääkemääräyskäytäntöjen ohjauksella ja lainsäädännöllä. Vuosina 2024–2025 useat avoterveydenhuollossa käytettävät biologiset lääkkeet tulevat apteekissa tapahtuvan lääkevaihdon (apteekkivaihdon) piiriin. Potilaiden näkemykset biologisista lääkkeistä ovat tärkeä tutkimusaihe hoitotulosten, lääkevaihdon, rationaalisen lääkehoidon edistämisen ja lääkepolitiikan kehittämisen näkökulmista. Tutkimuksen tavoitteena oli tutkia potilaiden näkemyksiä biologisten lääkkeiden hinnoista, kustannuksista ja niiden merkityksestä. Tavoitteiden mukaiset tutkimuskysymykset liittyivät: 1) potilaiden preferenssiin lääkkeiden hoidollisesta arvosta lääkkeen hintaan verrattuna (ensisijainen tutkimuskysymys) ja yhteiskunnan lääkesäästöistä, 2) lääkkeiden hinnan merkitykseen lääkevaihdossa (potilaiden taloudellisten taustatekijöiden vaikutus ja euro-määräinen hyväksymis/maksuhalukkuus lääkevaihdossa) ja 3) potilaan oman lääkehoidon kustannettavuuteen. Tutkimus perustui Yliopiston Apteekin (YA) ja Helsingin yliopiston (HY) tammikuussa 2021 toteuttaman kyselytutkimuksen aineistoon. Kyselyyn vastasivat YA:n kanta-asiakkaat sekä Reumaliiton ja IBD- ja muut suolistosairaudet ry:n viestinnän kautta tavoitetut henkilöt. Kysely oli kohdistettu reuma-, IBD- (tulehduksellinen suolistosairaus) ja ihopsoriasispotilaille, jotka käyttivät alkuperäistä biologista lääkettä (BA), biosimilaaria (BS) tai perinteisiä pienimolekyylisiä lääkeitä (PL). Vastaajia oli yhteensä 1338 (BA-käyttäjiä 226, BS-käyttäjiä 71 ja PL-käyttäjiä 1041). Tulosmuuttujina käytettiin yksittäisiä kysymyksiä ja summamuuttujia. Lääkekäyttäjäryhmän ja muiden taustamuuttujien yhteyttä tulosmuuttujiin tutkittiin kaksi- ja monimuuttuja-analyyseillä. Suurin osa (83 %) potilaista oli sitä mieltä, että lääkkeen hinta ei saisi vaikuttaa lääkkeen valintaan biologista lääkettä määrättäessä, ja 62 %:n mielestä biosimilaarien käyttö auttaisi säästämään terveydenhuollon lääkekustannuksissa ja mahdollistaisi suuremman potilasmäärän hoidon biologisilla lääkkeillä. Potilaan taloudelliset taustatekijät eivät olleet monimuuttuja-analyysin perusteella yhteydessä näkemyksiin biologisten lääkkeiden lääkevaihdosta tai kiinnostukseen lääkevaihdosta. Jos biologisen lääkkeen hypoteettinen omavastuuhinta potilaalle olisi 600 euroa vuodessa, 14 % alkuperäisvalmisteen käyttäjistä olisi valmis vaihtamaan biosimilaariin, jos sen kustannus olisi hänelle 30 % nykyistä pienempi. Biosimilaarien käyttäjistä 38 % olisi valmis maksamaan lisää saadakseen alkuperäisvalmisteen. Biologisten lääkkeiden käyttäjillä (BA 36 % ja BS 44 %) oli ollut enemmän taloudellisia ongelmia lääkkeiden ostossa kuin perinteisten lääkkeiden käyttäjillä (25 %) (p <0,001). Potilaat suhtautuivat yleisesti myönteisesti biosimilaarien käyttöön lääkekustannusten hillitsemiseksi, mutta pitivät hoidollisia perusteita hintaa tärkeämpänä. Potilaan taloudelliset tekijät eivät olleet yhteydessä näkemyksiin lääkevaihdosta tai vaihtohalukkuuteen. Merkittävä osa potilaista on kiinnostunut vaihdosta edullisempaan biosimilaariin. Tulokset korostavat biologisiin lääkkeisiin ja lääkevaihtoon liittyvän lääkeinformaation merkitystä.
  • Kiviluoto, Katrimari (2014)
    In a pharmacotherapy process prescriber, provider, administrator and evaluator of the effects of medication cooperate in a coordinated way to ensure optimum outcomes of the patient's medications. This requires that all professionals involved in the pharmacotherapy process know their functions and responsibilities in an interprofessional team. No previous studies have explored legislative facilitators and barriers that have impact on the interprofessional pharmacotherapy process even though interprofessional collaboration for assuring safe and effective pharmacotherapy is one of the main objectives of Medicines Policy 2020 in Finland (Ministry of Social Affairs and Health 2011). The aim of this study was to examine how legislation prevents or facilitates the interprofessional pharmacotherapy process. The data consisted acts, decrees, regulations and directives concerning pharmacotherapy process in the healthcare. Changes in the Finnish legislation from 1990 until present were studied, taking also into account the European Union legislation since 1995 when Finland became a member. Research method was qualitative content analysis. A modified form of the causal diagram of the impact of law and legal practices on public health system performance by Burris et al (2012) was used as a theoretical framework. Interprofessional collaboration applicable to pharmacotherapy process has been taken into account in the main enactments. These enactments concern rights and duties of healthcare professional, patient’s rights, resources required in healthcare, communication and information transfer, and pharmacotherapy process. Enactments that complicate the interprofessional pharmacotherapy process relate especially to communicational barriers. Facilitating enactments include guidelines ensuring coordinated clinical practice for effective and safe medication use and improved communication, collaboration and patient-centeredness as well as accessibility of cross-border healthcare. A trend was observed that the legislation enacted on the 21st Century put more emphasis on interprofessional collaboration, development of technology and mobility of citizens. According to this study there are more facilitators than barriers for interprofessional pharmacotherapy process. Recent enactments even emphasize more interprofessional cooperation in health care than did those enacted before the 21st Century. Still, enactments are general: they should be made more detailed to give a better understanding of interprofessional cooperation, also related to the pharmacotherapy process for safe, effective and evidence-based medication use.
  • Kaukovuori, Jouni (2021)
    Johdanto: Terveydenhuollon tietojärjestelmiin potilaista kertyvää tieto on hyvä esimerkki massadatasta. Se muodostuu lukuisista yksittäisistä, irrallisista tapahtumista. Potilastiedon toissijaisella hyödyntämisellä tarkoitetaan tiedon käsittelyä muuta tarkoitusta kuin potilaan terveyden edistämistä ja hoitamista varten. Toisiokäytölle on tyypillistä, että pääasiallisena kiinnostuksen kohteena ei ole yksilötason tiedon hyödyntäminen, vaan isommasta potilasjoukosta saatava summatieto, josta yksilön tunnistetiedot on poistettu. Toisiokäyttö mahdollistaa potilastiedon hyödyntämisen esimerkiksi tieteellisessä tutkimuksessa ja tietojohtamisessa. Tavoite: Tutkimuksessa selvitettiin HUSin tietoaltaasta louhitun aineiston avulla, onko tietoaltaaseen tallennetun aineiston avulla mahdollista tutkia lääkehoidon turvallisuutta ja rationaalisuutta sekä selvittää, millaisessa muodossa tietoaltaan data saadaan käyttöön ja millaisia toimenpiteitä datalle tulee tehdä, jotta sitä voidaan hyödyntää potilastiedon analysoinnissa. Aineisto ja menetelmät: Massadatan hyödyntämistä pilotoitiin rekisteritutkimuksessa, jossa esimerkkinä käytettiin opioideja. Rationaalisen lääkehoidon toteutumisen tutkimiseksi määriteltiin lääkeindikaattorit eli tunnusluvut, jotka oli tarkoitettu opioidien lääkehoidon kokonaiskuvan tarkasteluun. Indikaattoreiden avulla luotiin pohja hakuparametreille ja lausekkeille, joita tietoallashaussa käytettiin. Aineisto louhittiin tietoaltaasta maaliskuussa 2020 ja se muodostui opioideja koskevista lääkemääräysmerkinnöistä, jotka oli kirjattu potilastietojärjestelmään 1.1.2015-31.12.2019. Tulokset: Tietoallashausta saatiin 321 000 potilaan opioidimääräysdataa yhteensä noin 1,73 miljoonaa riviä. Kotiutumisen yhteydessä annetut opioidireseptit rajattiin jatkoanalyysin ulkopuolelle, sillä niitä koskeva tieto ei ollut rakenteisessa muodossa. Sairaalassa annettuja säännöllisiä opioidilääkemääräyksiä koskeva aineisto oli noin 258 000 riviä. Dataa siivottiin, järjesteltiin ja validoitiin data-analyysiä varten. Toimenpiteistä huolimatta data ei soveltunut indikaattorien laskentaan. Johtopäätökset: Tietoallasaineiston käytön mahdollisuudet rationaalisen ja turvallisen lääkehoidon tutkimukseen olivat tämän tutkimuksen perusteella rajalliset. Massadata-aineiston saattaminen tutkimuksellisesti hyödynnettävään muotoon vaatii menetelmän, joka pitää sisällään useita työvaiheita ja niiden kehittäminen vaatii tietoteknistä erityisasiantuntemusta. Vaikka dataa saatiin paljon, yksittäisen potilaan opioidilääkehoidosta ei saatu kokonaiskuvaa, koska merkittävä osa datasta oli rakenteettomassa muodossa. Potilastiedon toissijaisen hyödyntämisen kannalta aineiston rakenteisen osan merkittävimmät käytön esteet liittyivät datan laatuun ja luotettavuuteen. Jotta tietoaltaasta saatava aineisto soveltuisi toisiokäyttöön tai tieteelliseen tutkimukseen, pitää sekä potilastietojärjestelmän merkintä- ja kirjaamistapoja yhtenäistää sekä data tulisi tallentaa tietoaltaaseen yhä rakenteisemmassa muodossa.
  • Virolainen, Jenni (2012)
    Patient safety is a part of quality and safety of care. Patient safety is defined as freedom for a patient from unnecessary harm or potential harm associated with healthcare. Patient safety covers safety of care, medication safety and safety of devices. Different authorities have promoted patient safety in Finland. The Ministry of Social Affairs and Health set up the Steering Group for the Promotion of Patient Safety for a term extending from 1 November 2006 to 31 October 2009 to coordinate the work for promoting patient safety and to evaluate related development needs at the national level. It has published a national Patient Safety Strategy for the years 2009-2013. Patient safety is also included in the Health Care Act. The National Institute for Welfare and Health (THL) has a Finnish national programme on patient safety: Patient Safety with Skills. Other authorities promoting patient safety in Finland are Finnish Medicines Agency (Fimea) and National Supervisory Authority for Welfare and Health (Valvira). Many studies are related to Patient Safety. In the Seminar of Patient Safety Research 2011 studies were separated to following categories: Patient Safety Culture as a Challenge for Organisations, Medication Safety, Safeguard of Care, Preparation for Patient Safety, Learning of Patient Safety, Control of Patient Documents and Financing of Patient Safety. The aim of this study is to explore Finnish patient safety studies. This study was conducted by using an electronic survey. The survey was sent to members of Finnish Patient Safety Society and a mailing list of Patient Safety Network. The survey was also sent to attendees of the Seminar of Patient Safety Research 2011. Altogether 81 responses were obtained. A patient safety research had been done in 60 per cent of organizations. A patient safety research will be done in 62 per cent of organizations. 10 per cent of the researches were meant for the internal use of the organization but were also published in Finland and abroad. 21 per cent of the researches were published in Finland and abroad. 18 per cent of the researches were published only in Finland and 12 per cent only abroad. 25 per cent of the researches were meant only for the internal use of organization. 14 per cent of the respondents left this question unanswered. A personal grant from a foundation was the most common way of financing for patient safety research. Many different kinds of sponsors were also mentioned. There was co-operation between organizations in 58 per cent of researches. 86 per cent of respondents were interested in a network of patient safety researchers. Using of Reporting System for Safety Incidents in Health Care Organizations (HaiPro) was asked as a detail of this study. HaiPro was used in 65 per cent of organizations. 89 per cent of respondents said that their organization takes advantage of HaiPro but the level of use varied between respondents.
  • Eronen, Anna-Kaisa (2016)
    Medication safety meaning the safety of using medication is an important part of patient safety. Medication errors are the most common preventable threats of patient safety. Medication errors can occur in all stages of the medication process. Rather than blaming individuals involved in the process, incidents should be evaluated based on system thinking with an aim of identifying system and process-based weaknesses allowing errors to happen. James Reason's human error theory provides a good framework to investigate the topic from this perspective. The objective of this study was to gather information on the medication errors based on the Patient Insurance Centre 2013-2014 data of compensated medication errors. One of the aims was to identify different types of medication errors and gather information on their backgrounds and drugs involved. Another aim was to investigate the causes behind the medication errors and the views of the people involved on the contributin factors of these errors. Additionally the study aimed at identifying situations where interprofessional collaboration could have prevented medication errors from occurring. Finally the results of the study were also compared with the results of the earlier studies done using similar data. The data of the study consisted of 205 cases where medication error had caused compensated patient injury. Factors behind the errors were analyzed using descriptive statistics. The examples of most common cases were investigated more in depth through simplified root-cause analysis. Content analyses were used to gather information on the views of the people involved in errors as well as on the possibilities of preventing errors through interprofessional collaboration. The most frequent error type was omission of medication. The majority of the errors occurred in the early stages of the process when decisions on medication and treatment were done. There were altogether 250 drugs in the data out of which 98 different active ingredients were identified. Antithrombotic agents were the most common therapeutic group causing medication errors. 37% of all drugs included in data were classified as high alert medicines. More efficient use of the interprofessional collaboration could have prevented several medication errors. The number of medication errors had somewhat increased compared to earlier studies but the profiles of errors were very similar. The data of Patient Insurance Centre provides valuable information on medication errors across Finland. More accurate information on factors leading to medication errors could be obtained by improving voluntary nationwide reporting. This would make it easier to develop operating models that improve patient safety.
  • Erkkilä, Outi (2023)
    Physiologically based pharmacokinetic modelling (PBPK) can be used to predict pharmacokinetic behaviour of new drug molecules in human. PBPK model represents the body anatomically and physiologically with compartments connected to each other and combines those to drug specific parameters. PBPK modelling can be used to predict the absorption, disposition, and time-concentration profiles of drug molecules. The purpose of the study was to build a PBPK model for new drug molecule under research (compound A) and predict pharmacokinetics in human, to support the selection of dosing interval, formulation, and sampling time points for the first clinical trial. In this work it is described the building of the model in the ”bottom-up”-approach using in vitro parameters in GastroPlusTM-software. The modelling was done also for preclinical species (mouse, rat, dog) comparing the simulations to the observed in vivo data, which gave the confidence to the methods used in the modelling also for human. The model was first built for systemic kinetics and thereafter it was used for predicting pharmacokinetics after oral dosing. Parameters of systemic kinetics were compared also to the predictions from allometric scaling. Based on the preclinical species the most predictive method for the volume of distribution of compound A was the method by Lukacova, which predicted the volume of distribution to be moderate in human (1.7 l/kg). From the in vitro-to-in vivo -extrapolation methods the most predictive method to predict the clearance was the method by Poulin, which predicted low clearance in human (8.1-14.3 l/h). Empirical scaling factors based on the preclinical data were not needed, as the models predicted well the observed in vivo data. Allometric methods predicted the systemic kinetic parameters to be in the similar range. Advanced compartmental absorption transit -model (ACAT) integrated to GastroPlusTM-software predicted the absorption after oral dosing well in the preclinical species (predicted/observed ratio 0.8-1.3 for systemic exposure) despite the low solubility of the compound A. The model predicted the absorption in human to be sensitive to particle size and absorption rate to be clearly affected by the particle size. The feeding status was also predicted to affect on the absorption with larger particle sizes. The gut metabolism was not predicted to limit the oral exposure notably, whereas moderate bioavailability was predicted to be achievable. Compound A could be given in a capsule if the target particle size distribution could be achieved. The built PBPK-model can be used in the future to predict the first clinical doses by comparing the predicted plasma concentrations to in vitro pharmacodynamic parameters and to the plasma concentrations needed for efficacy in the pharmacodynamic models. The model can also be used to predict the drug-drug interactions.
  • Taivainen, Sanna (2016)
    Suspension is nowadays the most commonly used dosage form in preclinical animal studies. However, suspension can be physically unstable and changes in particle size or crystal form of an active pharmaceutical ingredient (API) can occur during storage. Conventionally suspensions are also prepared in a mortar, and hence the quality of suspensions is operator-dependent. One of the aims in this study was to prepare suspensions using a mortar and pestle and an Ultra-turrax homogenizer to find out how the preparation method affects the particle size of suspension. A solution containing methylcellulose and Tween 80 was used as a vehicle, and five active APIs with different physico-chemical properties as model drugs. Moreover, an aim of the study was to evaluate the stability of the suspensions stored at room temperature and in the refrigerator and freezer by physical (laser diffraction, optical microscopy, X-ray powder diffraction) and chemical (high-performance liquid chromatography) methods of analysis. The aim of the study was also to assess and compare the suitability of laser diffraction and optical microscopy for the determination of partice size during preclinical studies. The suspensions prepared using a mortar and pestle and Ultra-turrax had a similar particle size in almost all cases. The particle size of API that was difficult to grind decreased significantly, also when using Ultra-turrax although the capacity used was minimum. Both prepation methods had the best repeatability of particle size when the API was easy to grind. However, Ultra-turrax could provide better homogeneity of quality than a mortar and pestle if the settings were optimized. The effect of different operators was not studied in this study. The stability of suspensions in different storage conditions was dependent on the properties of API. The particle size of all frozen suspensions decreased after two days based on laser diffraction results. Although the reason was not found from literature or supplementary tests (particle size analysis of the vehicle and pH-measurements), freezing of suspensions should be treated with caution based on this study. The crystal structures of APIs remained stabile with the exception of typical disproportionation of the API salt. Suspensions were mainly chemically stabile in all conditions, but water-solubility of API seemed to decrease stability. The micellar solubilization of drugs was also observed. The best way to determine the particle size of preclinical suspensions proved to be the combination of laser diffraction and optical microscopy images. The microscopy images confirmed the validity of the size distributions measured by laser diffraction and provided information about e.g. particle aggregation. On the other hand, optical microscopy image analysis was not suitable method for particle sizing.
  • Haapalainen, Joonatan (2022)
    Traditional 2D cell cultivating vessels and experimental models cannot often simulate natural chemical and physical environment of different cell types. For example, availability of oxygen, chemical gradients, messaging molecules, fluid pressure, flow and surface topography are factors that may affect significantly in cell differentiation, growth, cellular structure, and metabolism. Modular bioreactors like Quasi-Vivo® -system can be used to simulate these factors. Liposomes are particles of phospholipid bilayer with aqueous space enclosed within. They can be modified in numerous ways, like loading them with hydrophobic and hydrophilic molecules, changing their transition temperature or coating them according to different needs. Doxorubicin is effective and widely used cytostatic agent, but when administered as a free drug it has often severe side-effects, like cardiotoxicity. Goal of this thesis is to determine appropriate manufacturing parameters and verify adequate shelf-life of ICG-Doxorubicin liposomes, that they are applicable for future in vitro experiments. Then survival of HepG2 cell line under flow in Quasi-Vivo®-equipment is determined, after which A549 and HepG2 will be then combined into one two-cell model. Finally, a simple illumination experiment in this cell model with previously made liposomes is conducted, and the effect in whole system is examined. Using protocol presented in this thesis it is possible to produce successfully and repeatedly liposomes with both ICG and doxorubicin encapsulation over 70%. Their shelf-life was at least 14 days when stored in 4°C protected from light. This was determined to be sufficient for in vitro testing. Cultivating A549 and HepG2 cell lines combined in the same system with shared media and fluid flow conditions was successful. Neither of the cell lines show significant difference in viability when compared to static control. When light-activating liposomes are administered to the system and then illuminated, from preliminary results we can see significant difference in drug effect. Both illuminated chambers and off-target chambers connected via Quasi-Vivo® show increased suppression, which shows promise that this in vitro model would be useful for future experiments.
  • Ollinkangas, Joni (2022)
    The problems caused by hypromellose in sterile filtration of ophthalmic products in the pharmaceutical industry were investigated. The research project was performed at NextPharma Oy's ophthalmics manufacturing facility in Tampere during the autumn of 2020. Hypromellose is an excipient commonly used in ophthalmic products as a viscosity enhancer to prolong the contact time of the preparation on the eye surface. In the ophthalmics compounding process, hypromellose is first dispersed by slowly sprinkling it into a hot solution and thoroughly mixing, after which the solution is cooled to room temperature. During cooling, the hypromellose dissolves and gels, increasing the viscosity of the solution. Incomplete dispersion or dissolution of hypromellose during the manufacturing process can slow down the filtration rate or even clog the filter completely due to undissolved hypromellose polymer material. Hypromellose is an industrially produced cellulose derivative that often contains some amounts of unreacted cellulose and other sparingly soluble polymer particles as impurities, which can also cause problems in filtration processes. Sterile filtration is a commonly used sterilization method for ophthalmic products, in which the prepared bulk solution is filtered through a 0.1 to 0.2 µm pore size filter membrane into a sterile receiving vessel. Due to the very small pore size, sterile filters are easily clogged if the solution contains poorly dissolved material. The purpose of this work was to collect additional information on the possible causes of clogging caused by hypromellose and to determine whether the filterability of a solution containing hypromellose can be improved by optimizing the manufacturing process parameters. The design of experiments was prepared, creating a two-level full-factorial test matrix without replicates and with three centre points. Four different process parameters were used (mixing time, mixing speed, dispersion temperature, and cooling temperature). Minimum and maximum levels for the parameters were obtained in the initial tests, after which the test solutions were prepared and filtered in a randomized order according to the test matrix. The aim of the screening was to find out which parameters were affecting the filterability and what would be their optimal combination that would maximize the filtration rate and the yield of filtration. Finally, the optimized parameters were used to test different batches of hypromellose, comparing the results to previous filtration tests. Additionally, an alternative hypromellose dispersion method was tested to minimize the amount of insoluble material remained during the dispersion and cooling steps. Of the parameters tested, mixing speed was the least significant, while cooling temperature had the most effect on the filtration results. The solutions with lower cooling temperature had better filtration results, which may be due to reduced aggregation of hypromellose due to increased hydration of the polymer chains. The temperature behaviour of hypromellose solutions could be an interesting subject for further investigation. Longer mixing times and higher dispersion temperatures produced slightly better filtration results on average, but the differences were not statistically significant. Most challenging in the study was controlling the temperature and mixing of the solutions, and the retention of insoluble hypromellose material at the walls of the compounding vessel. The alternative dispersion method gave promising preliminary results, but the method still requires further testing. It would be important to also find the root cause of the filter clogging mechanism e.g., by further analysing the clogged filter membrane. The study provided additional useful information of the behaviour of hypromellose solutions in solution preparations and during sterile filtration, which has been helpful in solving production problems.
  • Tiilikainen, Saija (2016)
    Prolyl oligopeptidase (PREP) is a serine protease which is extensively present in the mammalian system and especially abundant in the brain. Despite the long research history of PREP its physiological function has remained unclear. PREP has been suggested to regulate the functions of many bioactive peptides by hydrolysis and on the other hand to participate in several intracellular processes probably via direct protein-protein interactions. One of the functions suggested for PREP is the regulation of the brain neurotransmitter systems and based on, for instance, the location in the brain PREP has been connected to both excitatory and inhibitory neurotransmitter systems. The literature review of this thesis first describe the brain neurotransmitter systems associated to PREP in general with some examples of diseases related to their malfunctions. In addition the structure of PREP and its location in the brain, both subcellular and cellular levels, and in distinct neurotransmitter systems, are presented, after which the different proposed functions for PREP are reviewed. The aim of the experimental part of this thesis was to investigate the effects of PREP on the brain neurotransmitter concentrations in the mouse nigrostriatal pathway and also to mouse motor behavior. The main research methods were microdialysis, tissue assays and cylinder test. The study was composed of two sections with five week duration each. The first section was performed with wild-type mice expressing naturally PREP and the second section with PREP-knockout (ko) mice and their wild-type littermates. The mice were injected unilaterally above the substantia nigra with adeno associated (AAV1) hPREP viral vector or with AAV1-eGFP (green fluorescent protein) viral vector as a control treatment. The cylinder test was carried out before the injection, and two and four weeks afterwards. Microdialysis was used to study the actions of PREP on the extracellular concentrations of dopamine (DA) and its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), gamma-aminobutyric acid (GABA) and 5-hydroxyindoleacetic acid (5-HIAA), the major metabolite of serotonin (5-HT). In addition to the baseline assay the concentrations were measured after two amphetamine treatments (10 and 30 µM) administered via the microdialysis probe. The probe guide cannulas were inserted to mice striatums 1-2 weeks before the microdialysis measurement. In the end of the experiment the tissue concentrations of DA, DOPAC, HVA, 5-HT and 5-HIAA were measured from striatum and substantia nigra. Both the microdialysis and tissue sample concentrations were quantified with high performance liquid chromatography. In the first study section the PREP enzyme activity was also determined from striatum. Neither the complete deprivation nor over-expression of PREP in the nigrostriatal pathway had clear or consistent effects on the levels of neurotransmitters studied when compared to naturally occurring PREP expression. When comparing the differences between control treated groups of PREP-ko and littermate mice, a greater amphetamine stimulated DA-levels was seen in the former group proposing negative regulatory influence of PREP. In both study sections the tissue assay results were difficult to interpret due to observed responses also with AAV1-eGFP control treatment in comparison with untreated side of the brain. This was seen as a lower DA- and DOPAC-levels in substantia nigra and thus the meaning of the changes caused by PREP treatment is hard to comprehend. The results of the cylinder test may implicate some protective effect of the PREP-ko-genotype against viral vector injections in general. Then again the existence of compensatory mechanisms is possible when using knockout animals and thus the genotype differences are hardly ever unequivocal. The results of this thesis do not suggest outright regulatory effects of PREP on the neurotransmitter functions in the mouse nigrostriatal pathway although the confirmation of this requires further studies, especially in regard to GABAergic and glutamatergic systems. Studies should include a scale of different behavioral tests of motor activity and repeated microdialysis experiment with some defining method changes. The possible function and mechanisms of PREP as a regulator of neurotransmitter intake or release is rationale to study at molecular level with applicable methods.
  • Penttinen, Anne (2010)
    Prolyl oligopeptidase (POP, E.C. 3.4.21.26) cleaves short peptides, of less than 30 amino acid long, at the C-side of an internal proline. It has been associated with many pathophysiological processes, such as neurodegeneration and inflammation. At the moment there are no studies that have been focused on POP function in multiple sclerosis (MS). A preliminary study in a Spanish cohort reported altered POP activity in plasma samples of patients with relapsing-remitting multiple sclerosis (RR-MS) compared with healthy controls. Also they observed increased levels of the endogenous POP inhibitor in plasma samples of patients with RR-MS. The first objective of this study was to evaluate the POP activity levels in serum and cerebrospinal fluid (CSF) samples from RR-MS patients and healthy controls in a Finnish population using a kinetic fluorescence assay. The seral levels of the endogenous POP inhibitor were also investigated by preincubating recombinant porcine POP (rPOP) with serum and determining the percentual decrease of POP activity compared to basal rPOP values (inhibitory capacity %). The second objective of this study was to purify and characterize the endogenous POP inhibitor in serum. In order to accomplish this goal, different biochemical and biophysical features, such as temperature resistance and filtering cut-off were tested. Also a combination of chromatographic approaches (affinity/anion exchange/hydrophobic interaction chromatography) with polyacrylamide gel electrophoresis and protein staining was used. All the differences observed in POP activity/inhibitor levels (serum, serum with DTT, CSF) between healthy controls and patients with RR-MS in this study did not reach statistical significance due to low values in all the samples. However, the trends in all the measured parameters were similar to the preliminary study in a Spanish cohort. Thus, the data supports further, more comprehensive, studies on the role of POP in MS. After series of chromatographic runs, a mass spectrometry analysis revealed the endogenous POP inhibitor to be α2-macroglobulin, a panprotease inhibitor in serum. α2-Macroglobulin has also been associated with MS, thus this finding substantiate the relationship between POP and MS.
  • Kolu, Anna-Maija (2013)
    Spray drying is one way to dry protein medicines and it has many advantages compared to other drying methods, for example it is a fast process. In spray drying high temperature and mechanical stress can inactivate the protein. Disaccharides are generally used as protective agents of protein in spray drying because they have an ability to protect the structure of the protein during drying and storage. Aim of this research was to study the stability of the protein during spray drying and storage by using β-galactosidace as a model protein. Aim was also to characterize the physical properties of trehalose and melibiose and to study how well they protect the protein. Some of the central matters to be examined were the glass transition temperature, crystallinity, water activity, yield of the spray dried powder and protein activity. Especially studying the properties of melibiose in spray drying was important because it has not been used before. The study also included the optimization of the process parameters to be suitable for the product. Trehalose and melibiose transformed to an amorphous form during spray drying. Both XRPD and DSC showed an amorfous form. Trehalose and melibiose proved to be good protective agents for the protein during spray drying and storatge probably because they remained their amorphous structure. β-galactosidase remained activity very well. Optimizing of the process parameters was successful because protein remained its activity and still the powder was quite dry and yield was good. The changes in the structure of the protein were studied with FT-IR but the amount of the protein was too small. Problems caused by the spray drier may have an effect to the results, but on the other hand the spray dryer was made to work optimally.
  • Hovi, Marianne (2012)
    The burden of diabetes is increasing globally as the number of people with diabetes reaches over 220 million. Over 90 per cent of these people are suffering from type 2 diabetes. This condition is primarily defined by the chronic increase in blood glucose level or hyperglycemia. Type 2 diabetes is characterized by insulin resistance and is usually associated with abnormal insulin secretion. Insulin resistance is a state where normal amount of blood insulin is inadequate to increase glucose uptake in the most important target tissues of insulin. Numerous reports demonstrate that oversupply of lipids leads to loss of insulin activity and the formation of type 2 diabetes. Protein kinase C (PKC) isozymes comprise a family of serine/threoninekinases, which have a regulatory role in a multiple cellular processes. PKC!-isozyme activity is known to play a role in insulin resistance and therefore in type 2 diabetes. Free fatty acid (FFA) induced insulin gene function inhibition is associated with phosphoinositide dependent kinase1 (PDK1) independent phosphorylation of PKC!-isozyme in the most important insulin target tissues. Phosphorylated PKC!-isozyme causes insulinreceptor gene expression inhibition. Present study is part of a VHH-antibodies related research where the goal is to characterize these antibodies and to find out their effects on protein kinase C. VHH-antibodies are Ilama derived antibodies which contain a single heavy-chain variable domain, that is fully capable of antigen binding. In this work, we studied VHH-antibodies binding to PKC!-isozyme and its functional domains. PKC!-isozyme and its domains were produced in Sf9-insect cells. The binding was studied using Western blot and immunoprecipitation assays. In addition, the binding of 368 VHHantibodies to PKCε-isozyme's domain 2 were studied. With Western blot, it was discovered that E7-VHH-antibody binds to PKCε-isozyme full length and to domain 3. Other VHHantibodies tested in Western blot did not bind to PKCε-isozyme. Seven VHH-antibodies bound to PKCε-isozyme in immunoprecipitation. All of these VHH-antibodies bound to the full length and to domain 3, but not to other domains. In radioligand binding assays none of the VHH-antibodies bound to domain 2 that is the binding site to the endogenous PKCε-isozyme activator diacylglycerol (DAG). The results gathered with these three different methods were in line with each other. As the results gained from Western blot and immunoprecipitation show, all the VHH-antibodies, that bind to PKCε-isozyme, bind to its domain 3. With this study, we succeeded to gather new information about the binding of VHH-antibodies to PKCε-isozyme and its domains. The exact binding site has not been studied with so many VHH-antibodies before this study. Moreover, we also exploited methods that have not been used in this context before.
  • Heikkilä, Aki (2015)
    Lead molecule search is the first part of drug design. This process can be done using computerized docking of ligands into target proteins. Usually this requires expensive software and powerful computer systems specifically made for the process. There are however some programs that are available for free and can be run on home computers. The purpose of this Master's Thesis was to see how these free software can be used for the task of docking and also to create a method or a guideline for such work. Protein kinase C (PKC), a popular target for drug design, was chosen as a target of inhibitor design. PKC is part of a larger family of serine/threonine kinases and formed of 10 isoforms all with different effects on cellular functions. The large amount of related kinases and the similarities in their sequences make finding selective inhibitors a difficult process. Homology models of all PKC isoforms in three known conformations solved by x-ray diffraction (pdb: 1XJD, 2I0E and 3A8W) were created using Modeller. Into these models a set of possible ligands from the free database of molecules ZINC was docked using Autodock Vina utilizing a script created for docking multiple ligands into multiple targets. The dockings resulted in some interesting results. Six molecules were recognized as possible lead molecules for further research. None of these molecules had any patents or previous results of protein kinase inhibition connected to them. The most interesting result was the finding that coluracetam, a nootropic drug of the racetam family, might be a protein kinase inhibitor. Racetams are usually considered drugs that lead to PKC activation. It has been proposed that inhibitors may prolong the lifetime of kinases in the cells leading to increased activity in the long term. In our opinion coluracetam might prove to be a good tool for studying the complex way kinase activity is modulated. The methods and scripts used in this work will be released for free use.
  • Itkonen, Jaakko (2014)
    Proteins are endogenous molecules that carry out most biological functions in vivo. They are called as the biological workhorses. Proteins are made up of polypeptide chains that usually fold in the three dimensional space to adopt a native stable conformation. Stability of proteins is dependent on the interplay of environmental factors (pH, temperature, ionic strength). For most proteins, the biological function closely relates to the structural attributes of the protein. Misfolding or unfolding of proteins often result in aggregation. Protein aggregation in vivo is known to cause debilitating and fatal diseases such as Alzheimer's, Huntington's, Parkinson's and age related macular degeneration (AMD). Instability (physical and chemical) of proteins in vitro is believed to result in aggregation. This is a huge concern for the biopharmaceutical industry as it not only limits the effectiveness of the manufacturing process but also poses a great risk of fatality in vivo due to the immunogenic nature of the aggregates. Mechanisms of protein aggregation are complex and not well understood. Regulatory requirements for patient safety in biopharmaceutical products require characterization and analysis of aggregates in protein drug formulations. This review provides an overview of protein aggregation in general and highlights the different analytical methods used to characterize protein aggregates in biopharmaceuticals. Neurotrophic factors influence survival, differentiation, proliferation and death of neuronal cells within the central nervous system. Human ciliary neurotrophic factor (hCNTF) has neuroprotective properties and is also known to influence energy balance. Consequently, hCNTF has potential therapeutic applications in neurodegenerative, obesity and diabetes related disorders. Clinical and biological applications of CNTF necessitate a recombinant expression system to produce large amounts of functional protein. Previous studies have reported that recombinant expression of CNTF in Escherichia coli (E. coli) was limited by low yields and the need to refold the protein from inclusion bodies. In this report, we describe a strategy to effectively screen fusion constructs and expression conditions for soluble hCNTF production in E. coli. Most conditions tested with the codon optimized hCNTF sequence in fusion with soluble tags resulted in soluble expression of the protein. The construct 6-His-CNTF showed soluble expression in all the conditions tested. Our results suggest that codon optimization of the hCNTF sequence is sufficient for soluble expression in E. coli. The recombinant hCNTF was found to bind to CNTFRα with an EC50 = 36 nM.