Browsing by master's degree program "Master's Programme in Pharmacy"

The package leaflet (“leaflet”) is a technical document included in medicine packages to provide information about the medicinal product to the user. With the EU now encouraging the adoption of eHealth, it can be assumed that written medicine information would be included in the digitalisation process. Medicine users’ views on electronic forms of medicine information should be assessed before any changes can be made, but so far there is very little data on this. The aim of this study was to find out what kind of leaflet medicine users would prefer and how they would feel about an electronic leaflet. The main aim was to find out if there is a difference in preferences between different types of medicine users and between medicine users of different ages in the provision of a package leaflet. The study also sought to find out if the current leaflet is being read by medicine users. This study was conducted by carrying out a survey to pharmacy customers over the age of 16 collecting prescription medication(s) for themselves (n = 110). The data was collected at one retail pharmacy in Helsinki, Finland during July 2020. The data was analysed quantitatively. This study found that medicine users generally feel positively about an electronic leaflet (liked by 63%) and many are open to idea of an electronic leaflet (75%). The majority (88%) could see positives in using an electronic leaflet, regardless of leaflet preferences. The study did not find a difference between new and repeat medicine users in the preference for a particular leaflet format, but age is correlated with the preference for a particular leaflet type, with younger medicine users wanting an electronic leaflet as often as older medicine users want a printed leaflet. Having the leaflet appear in My Kanta pages after the medication has been dispensed was found to be the most popular way to receive an electronic leaflet. This study also found that there is a difference between new and repeat medicine users when it comes to reading the leaflet after a medication has been dispensed. With the current printed leaflet 81% of repeat medicine users and 38% of new medicine users do not read it. The most common reason given for not reading the leaflet was that the participant had read it before and did not feel the need to read it again. According to this study, medicine users, especially younger medicine users, feel positively about the idea of an electronic leaflet, which is encouraging for the future of an electronic leaflet. The results are in line with prior research, but also suggest that more medicine users feel positively about the idea of an electronic leaflet than before. The leaflet reading behaviours of medicine users also highlight the need for a system, where a medicine user can be alerted to any changes in the leaflet, which is something only an electronic system could do.

Heart failure is a global health issue that can result from various factors, one of which is myocardial infarction. The adult human heart has limited regenerative capacity to cover the loss of cardiomyocytes after myocardial infarction with new cardiomyocytes. The main responses to the loss of cardiomyocytes are fibrotic scar formation and the hypertrophy of remaining cardiomyocytes. Prolonged hypertrophy eventually leads to heart failure. Current treatments for heart failure only relieve the symptoms. Inducing cardiac regeneration could be one possible way to prevent and treat heart failure. Thus, to develop medical treatments that enhance the regenerative capacity, a comprehensive understanding of precise cellular mechanisms behind heart regeneration is crucial. The objective of this study was to establish a high-content analysis method for human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) utilizing the Cell Painting assay to identify and categorize morphological alterations induced by various compounds in hiPSC-CMs. To evaluate the morphological impacts, dozens or even hundreds of cell features were measured at the same time. hiPSC-CMs were exposed to two hypertrophy inducers, endothelin-1 and angiotensin II, and to doxorubicin, which is known to be a cardiotoxic compound. In addition, the effects of a GATA4- targeting compound, C-2021, on hiPSC-CMs were examined. C-2021, was expected to have antihypertrophic effect on the cells. Previously used methods, proBNP staining and qPCR, were used to validate the novel method. According to proBNP staining and qPCR, endothelin-1 induced cardiomyocyte hypertrophy greater than angiotensin II. Compound C-2021 did not show statistically significant antihypertrophic properties after hypertrophic stimuli, but some tendency the alleviate the hypertrophy was noticed. Moreover, by utilizing different data processing programs a novel analysis method was developed. With this method, the different treatment groups were clustered based on the morphological alterations caused by compounds exposures. The hiPSC-CMs exposed to endothelin-1, angiotensin II or doxorubicin showed a different morphological profile compared to the control group hiPSC-CMs. Compound C-2021 was also observed to affect cell morphology. However, the data analysis still needs improvements in order to detect which cell features these compounds affect.

Objectives To investigate distribution and causes of drug shortages in five selected countries with data from public shortage notification registers. Design Statistical retrospective analysis of national drug shortage registers Data Public shortage notification register data from the first nine months of 2020 in Finland, Sweden, Norway, Spain, and the United States, partly combined with national drug registers. Results Altogether 5132 shortage reports from Finland (n=1522), Sweden (n=890), Norway (n=800), Spain (n=814), and the United States (n=1106) published during the first nine months of 2020 were found in the study. More than half (54%) of the active ingredient level shortages (classified by ATC code) found occurred in only one country, and only 1% occurred in all five countries. On a country level, 19-41% of the shortages were found only in a given country. The distribution of shortages by ATC category and drug form was significantly different between countries, especially between the US and European countries. Injectables were found to have an especially high shortage risk in the US, much less so in European countries. On the other hand, some drug classes were in shortage almost exclusively in Europe but not in the US. Conclusions Although drug shortages are a growing global problem, drug shortages are rarely global, but typically occur only in some countries, while other countries have an uninterrupted supply of the same drug. Drug shortages should be seen not just as a problem of manufacturing disruptions, but as a question of equitable and effective distribution of drug supply on an international level. Price differences and other commercial issues could be factors behind variation of shortages found between countries.Considering the limited and probing nature of the study, further research of shortage register data is certainly warranted. International comparative register study is a meaningful and valid method for further understanding in this field.

The OATP1B3 belongs to the organic anion transporting polypeptides (OATPs) encoded by the SLCO (solute carrier organic anion) genes which belongs to the SLC (solute carriers) gene superfamily. It is an influx transporter which is primarily expressed on the basolateral membrane of the hepatocytes. It transports many endogenous substrates as well as clinically important drugs such as thyroid hormones and statins into hepatocytes and thus participates in the first step of hepatic metabolism. Single nucleotide polymorphisms (SNPs) of the SLCO1B3 gene can affect the pharmacokinetics and pharmacodynamics of its substrates. The aim of this study was to set up and optimize an in vitro method to study the function and expression of the OATP1B3 transporter and its genetic variants. SNPs 334T> G (Ser112Ala), 699G> A (Met233Ile) and 767G> C (Gly256Ala) and stop codon TAA were introduced into the SLCO1B3 gene by site-directed mutagenesis. Recombinant baculovirus vectors containing the genetic information of OATP1B3 and its variants were used to transiently transfect the HEK293 cells. After optimizing the substrate incubation time and concentration, as well as the viral load and selecting the fluorescent substrate (8-FcA), the uptake assay was used to determine the transport activity of the OATP1B3 variants in HEK293 cells. The transport activity of the artificial WTP variant was also investigated in this study. The transport activities of the Ser112Ala, Met233Ile and Gly256Ala variants did not change significantly from the wild type although the transport activity of the Met233Ile variant appears to be slightly impaired. In turn the WTP variant was unable to transport 8-FcA. Based on this study the function of OATP1B3 variants can be studied using recombinant baculovirus to transiently transfect the HEK293 cells. 8-FcA can be used as a probe substrate in these studies. The results of this study confirm previous knowledge of the functioning of Ser112Ala, Met233Ile and Gly256Ala variants. More studies are needed about the effects of these variants on the transport of OATP1B3 drug substrates. Also studies about the location, cell membrane and total cell expression of the WTP variant are needed to evaluate reliably the reasons of its inactivity.

Evidence based medicines alongside with age-appropriate dosage forms are essential in enabling appropriate treatment for any patient group. Pediatric pharmacotherapy, however, is lacking age-appropriate dosage forms and research-based evidence regarding the dosing, efficacy, and safety of medicines. Orally administered drugs require manipulation to enable administration and are often used against the indications approved in the marketing authorization and summary of product characteristics (SmPC). This off-label use puts pediatric patients at risk for medicational errors and adverse drug reactions. The aim of this study was to investigate recent trends in oral dosage forms used in pediatric randomized controlled trials (RCTs), with emphasis on age appropriateness. The results could be utilized in developing evidence-based dosage forms, suitable for all pediatric patients aged 0-17 years, and manufacturable in a small scale in a hospital pharmacy. This study was conducted as a systematic review following the PRISMA Statement. The literature search was carried out from Pubmed and Scopus databases and it covered a five-year period of 2015-2020. References from the databases were entered to the Covidence systematic review platform. After removing duplicates 3333 articles were left for screening. Two independent researchers selected the articles first screening by title and abstract, and then by full text review. A qualitative content analysis was conducted on the included articles. Altogether 77 articles met the inclusion criteria. Dosage forms included were tablets (n=37), liquids (n=21), capsules (n=18) and multiparticulates (n=6). Majority of the dosage forms were conventional (n=49) compared to more advanced novel modified release and fixed-dose combination formulations (n=33). Based on our results, orally administered dosage forms used in the recent pediatric RCTs are still limited by poor acceptability, palatability, and the need to manipulate dosage forms to enable administration. These issues are similar to the ones related to the off-label use of medicine that compromise patient safety. Majority of the dosage forms included in our study were tablets, indicating a positive shift towards more safe and acceptable dosage form. Formulations were also evolving towards dispersible, extended-release and fixed-dose combinations that offer additional benefits for pediatric patients. The low number of children < 2 years old included in study populations and the poor acceptability profile reported by most studies limit our conclusions on an ideal age-appropriate dosage form. Further research is needed on unifying the guidelines used in pediatric drug development.

P-glycoprotein is an efflux transporter of the ABC family. It is expressed mainly in tissues that have a role in limiting the absorption and distribution of xenobiotics in the body or their elimination. P-glycoprotein is known to have an important role for example in the blood-brain barrier and in protecting the fetus from xenobiotics in the mother’s blood stream. Genetic polymorphisms in transporter proteins can cause individual differences in the pharmacokinetics of drug substances, which can lead to differences in drug efficacy or side effects. In the ABCB1 gene, which codes for p-glycoprotein, several polymorphisms have been discovered. The frequencies of these polymorphisms vary in different ethnic populations. Previous studies have shown that the effects of these polymorphisms are often substrate-dependent. Since there are several confounding factors usually present in clinical association studies, in vitro studies are needed to clarify the effects of individual polymorphisms. Polymorphisms can be studied in vitro by making intentional mutations to the gene sequence and expressing the variant gene in a suitable cell line. In this study four variant p-glycoprotein genes (c.781A>G, c.1199G>T, c.2005C>T and c.3421T>A) were created by site-directed mutagenesis, and expressed in HEK293 cells using a baculovirus recombinant protein expression method. The effects of the polymorphisms were studied by determining the expression level and the transport acitivity of the variant proteins compared to the wild-type. Western blot was used to determine the expression level and a calcein accumulation assay in HEK293 cells was used to compare the transport activities. Also a membrane vesicle transport assay with n-methyl quinidine was set up and optimized, but the variants were not yet studied with this method during this study. In this study no statistically significant differences were found in the transport activities of any of the four variants compared to the wild-type p-glycoprotein. Also the differences in protein expression level between wild-type and variant proteins were small. However, because of the previously reported substrate dependency of polymorphism effects, it would be beneficial to study the variants with at least one other substrate and one other assay method, and thus the membrane vesicle transport assay would be useful to further compare the transport activities of variant proteins to the wild-type p-glycoprotein.

Cardiomyocyte oxygen deprivation followed by apoptosis and cardiomyocytes being replaced with fibrotic tissue can lead to heart failure. Cardiovascular diseases are the most common cause of death world-wide, contributing to 17.8 million deaths in 2017. Treatments currently available aim to maintain cardiac function but are unable to repair the damage, resulting in a poor prognosis for heart failure. Cardiomyocytes are able to proliferate but the endogenous mechanisms of cardiac repair are insufficient to replace the damaged cardiomyocytes, as only an estimated 0.3-1 % of adult cardiomyocytes are regenerated annually. It is known that before birth and up to seven days after birth mice can maintain ability to regenerate cardiomyocytes even after large damage, but this capability is lost within seven days following birth. After this, cardiomyocytes exit cell cycle and will not re-enter it sufficiently to enable cardiac repair. In adults the growth of heart muscle results mainly from hypertrophic growth meaning that the cells grow in width and length. Cardiomyocyte regeneration is an important therapeutic target to which there are no effective pharmacological therapies available yet. The aim of this study was to investigate the effect of 14 novel compounds on cardiomyocyte viability, phenotype and cell cycle activation. Novel compounds were synthesized at the Faculty of Pharmacy, Division of Pharmaceutical Chemistry and Technology, University of Helsinki in Finland. Initial toxicity and cell viability screening was conducted with lactate dehydrogenase assay (LDH assay) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT assay) using COS-1 cells. Based on these assays tolerable concentrations of compounds were determined. Activity analysis was conducted using human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) and immunocytochemistry staining in conjunction with high-content analysis (HCA). Stress response was measured by imaging and analyzing expression of pro-B-type natriuretic peptide (proBNP) and cell cycle activation was imaged and analyzed by using 5- bromo-2’-deoxyuridine (BrdU) as a marker of active cell cycle. In addition, the DNA content of the cardiomyocytes was measured using 4’,6-diamidino-2-phenylindole (DAPI) staining as well as cardiomyocyte morphology investigated with cardiac troponin T (cTnT) staining. One of the compounds, K6, decreased proBNP expression, which can be considered as a sign of decreased stress response. However, K6 also decreased the number of BrdU positive cardiomyocytes that can be considered as a sign of decreased cell cycle activity. Together these markers indicate that the decreased activity may not be due to a stress response caused by the compound. Another compound, K12, increased proBNP expression in all tested concentrations and it also decreased the number of BrdU positive cardiomyocytes. Together these could be considered as an indication of cardiotoxicity. The rest of the compounds did not exhibit remarkable biological activity or there was too great variance between the results of the independent experiments (n=3) to draw definite conclusions. Compounds for this study were chosen for the sole reason of not been tested for biological effects before. Using a defined compound library or screening a larger number of compounds could deliver more predictable results. Early toxicity and viability screenings were a good approach allowing to define toxic compounds and concentrations before continuing with further studies. Pharmacological therapies to induce cardiac regeneration will continue to be an important area of interest in cardiovascular drug research. Phenotypic screening in conjunction with high-content analysis offers variable and statistically significant data on cardiomyocyte proliferation and stress response. The results of the screening could be improved with careful selections of test molecules based on their structure and biological activity. Early toxicity and viability screening further improve the predictability of results. As a result of this study a compound that would induce cardiomyocyte proliferation was not found, however, one compound that seemed to decrease cardiomyocyte stress response was detected and this compound could be of interest for further studies.

Solunulkoiset vesikkelit eli EV:t ovat nanokokoisia solujen tuottamia lipidikaksoiskalvon peittämiä kalvorakkuloita. Solut vapauttavat EV:itä solunulkoiseen tilaan ja niitä on kaikissa kehon nesteissä. Aiemmin niiden uskottiin olevan vain solujen tapa päästä eroon tarpeettomasta materiaalista, mutta nykyisin tiedetään, että EV:illä on tärkeä merkitys solujenvälisessä viestinnässä. Sitä mukaa kun ymmärrys EV:iden merkityksestä on kasvanut, on kasvanut myös kiinnostus niiden tutkimiseen. EV:itä voidaan eristää lähes kaikista kehon nesteistä, mutta veressä niitä on erityisen runsaasti. Plasman EV:t ovat pääosin peräisin punasoluista ja verihiutaleista. Kun nanopartikkelit ovat kosketuksissa veren kaltaisten biologisten nesteiden kanssa, niiden ympärille muodostuu proteiinirakenne, jota kutsutaan proteiinikoronaksi. Proteiinikoronan koostumus vaikuttaa nanopartikkeleiden pintaominaisuuksiin. Se voi vaikuttaa myös esimerkiksi niiden soluinteraktioihin ja signalointiominaisuuksiin. Tämän pro gradutyön tarkoituksena oli tutkia punasolujen ja niistä tuotettujen nanoerytrosomien EV tyypin proteiinikoronan määrää ja vertailla näitä määriä toisiinsa. Mittaukset suoritettiin ihmisen veri plasmasta, joka oli pitoisuudeltaan 100 %:sta, 50 %:sta sekä 25 %:sta. Verestä peräisin olevien EV:iden etu sekä mahdollisina lääkekuljettimina, että tutkimuskäytössä on se, että ne ovat myrkyttömiä, heikosti immunogeenisiä, helposti saatavissa olevia, helppokäyttöisiä sekä varastoitavia. Tutkimustulosten perusteella proteiinikoronan määrä on EryEV:illä ja NanoEry:illä samaa suuruusluokkaa. Havaittavaa eroa ei ainakaan näin pienellä otoskoolla ollut havaittavissa.

Background. Critical thinking skills, which are an aspect of generic skills, are related to the success students have in their university studies and working life after graduation. When it is considered whether the current degree programs and education systems support sufficiently the development of the skillset needed in the future working life of a university student, one can focus on examining the development of critical thinking skills of a student. Due to the lack of scientific studies on critical thinking, research should be done on the field of pharmacy to gather information on the critical thinking skills of master of pharmacy students and to determine, if the master’s degree program in pharmacy is generating the kind of professionals needed in the working life. Aim. The main goal is to study the level of critical thinking skills possessed by students in the beginning of their Master of Pharmacy studies. Combining the theoretical background and empirical study is considered in choosing a suitable method for the study. The method chosen for the study has to be applicable to determining a student’s ability to interpret, analyse and evaluate information, based on which a student consciously arrives on a conclusion while taking into consideration the context and different point of views of students. In addition to studying critical thinking skills, the aim is to analyse the students’ quality of argumentation and the use of references in comprising quality arguments. Methods. The studied set of students comprised of 17 students on their first year in the Master of Pharmacy program in University of Helsinki. The study was performed using a constructed-response task which was based on a real-life problem and was composed of three statements the students had to address utilizing the provided materials. When answering an open question, a student has to be able to analyse, evaluate and synthesize information from different sources. Based on this process, the student must demonstrate in their own words the relevance of the processed information and to construct a coherent essay that also utilizes the provided sources appropriately. The critical thinking skills of the students were evaluated from three aspects: the relevance of the content, processing of information and augmentation. A table for evaluation and classification of the data was created based on the literature study on critical thinking. According to the table, the studied material was divided into three categories. Category 1 entailed students with essays showing the lowest level of critical thinking skills while category 3 entailed students with essays showing the highest level of critical thinking skills. Results and conclusions. A deficiency in the level of critical thinking and argumentation skills possessed by the students was found, which has been encountered in previous studies, as well. All in all, only three out of the studied 17 students, showed impressive critical thinking skills. Most of the student (82%) showed defective argumentation, and more than half (65%) had problems with referencing. In addition, all but one student struggled in utilizing the information gathered from the reference materials. Based on the results, it is important to focus on developing the critical thinking skills of the students in the master’s program in pharmacy. To further study the matter, the same study should be performed on students at the end of their master’s studies to determine if the teaching and evaluation methods in the master’s program in pharmacy are sufficient in developing adequate critical thinking skills

Light-sensitive liposomes have gained attention for their ability to deliver cargo to tissues, offering spatiotemporal control over drug release. Red-light wavelengths have been utilized as an external trigger in light-sensitive reactive oxygen species (ROS)-mediated drug delivery, due to their favorable properties, such as the low light absorption by tissue chromophores. The ROS-sensitive drug delivery systems use photosensitizers (PS), which upon light exposure generate ROS in the presence of molecular oxygen. Palladium(II)phthalocyanine (Pd(II)PC), a new second-generation photosensitizer, can upon light irradiation generate relatively high singlet oxygen concentrations, enabling the efficient oxidation of the unsaturated lipids. The oxidation of the lipids leads to the disruption of the liposome bilayer and eventually, the release of the encapsulated cargo. To gain deeper insight on the phthalocyanine-labeled liposomes in drug delivery, a red light-triggered cationic liposome formulation encapsulating Pd(II)PC was formulated. The characteristics of the liposomes, the release mechanisms, and the release quantities of calcein (623 Da) and fluorescent-conjugated dextrans (4 000-70 000 Da) were studied following red-light exposer with 630 nm, 450 mW/cm2 laser while utilizing varying Pd(II)PC-loading quantities. Following oxygen removal and temperature-induced release studies, the mechanism of release of the liposomes was principally observed to be light-triggered reactive oxygen species-mediated. In the light-induced release studies an effective release of the calcein, and a relatively effective release of the Rhodamine B dextrans (10 kDa, 70 kDa) were observed from the liposomes via the Pd(II)PC-generated and reactive oxygen species-mediated oxidation of the unsaturated lipids. The release of the biomacromolecules from the liposomes was observed to require longer irradiation times than that of calcein. The longer irradiation times likely lead to deeper oxidation of the unsaturated phospholipids, resulting in a comprehensive eruption of the liposome bilayer. The comprehensive eruption of the liposome bilayer eventually enables the sufficient release of biomacromolecules from the liposomes.