Faculty of Pharmacy

 

Recent Submissions

  • Eriksson, Veronica (Helsingin yliopisto, 2020)
    Migrän rankades som den näst största orsaken till funktionshinder år 2016 i Global Burden of Disease (GBD) -studien. Människor med migrän har större funktionshinder och en lägre hälsorelaterad livskvalitet än befolkningen i allmänhet. Många migränpatienter upplever funktions- och känslomässig nedsättning på grund av sin sjukdom. Migrän kan begränsa deras dagliga aktiviteter och påverka deras privata, yrkesmässiga och sociala liv. Migrän påverkar patienten också mellan attackerna och kan påverka deras utbildning, karriär, familj samt nära och kära. Komorbida sjukdomar och misslyckade behandlingslinjer ökar migränbördan. Utöver dessa så medför migrän även en ekonomisk börda. Stigma beskrivs som den dolda sjukdomsbördan. Kroniska migränpatienter har visat sig ha högre stigma än episodiska migränpatienter. Även om migrän är en av de vanligaste funktionshindrande huvudvärkstörningarna, är den fortfarande både underigenkänd, underdiagnostiserad och underbehandlad. Syftet med denna studie var att fastställa omfattningen av migränbördan och migränstigman hos vuxna finländska migränpatienter. Studien siktade på att producera omfattande och aktuell information om migrän och dess svårighetsgrad i Finland, med betoning på den börda det medför för migränpatienterna och samhället. Migrän är vanligast bland befolkningen i arbetsför ålder, vilket ökar den samhälleliga bördan av sjukdomen. Denna studie genomfördes som en tvärsnittsundersökning med hjälp av en elektronisk enkät bland vuxna finländska migränpatienter. Deltagarna kontaktades via Migränföreningen i Finland. Frågeformuläret bestod av den redan existerande och validerade MIDAS-enkäten och av frågor som utvecklats av författaren. De slutliga datan bestod av 608 svar. Av alla respondenter med 8 eller fler huvudvärksdagar i månaden kategoriserades över 90% i gruppen för svår funktionsnedsättning (MIDAS grad IV) och hade därmed liknande funktionshinder som de med 15 eller fler huvudvärksdagar i månaden (dvs.respondenter med trolig kronisk migrän). Andelen respondenter med svår funktionsnedsättning (MIDAS grad IV) var större i denna studie (65,0%) än i en studie som genomfördes i Finland år 2000 (47%), vilket tyder på att funktionshinder på grund av migrän i Finland kan ha blivit svårare under de senaste två decennier. Den genomsnittliga nivån av huvudvärkssmärta i den aktuella studien var 6,2 (på en skala från 0-10) och smärta var den aspekt som de flesta respondenter betraktade som den värsta aspekten av migrän. Detta understryker vikten av korrekt smärtlindring vid migränvård. Många av respondenterna löpte också risk för överanvändning av läkemedel vilket betonar vikten av att övervaka läkemedelsanvändningen och informera patienterna om möjliga risker. Stress rapporterades som den vanligaste migränutlösaren, och att minska på stress på arbetsplatsen rapporterades också som det viktigaste sättet för hur migrän bättre kunde hanteras på arbetsplatsen. Nästan hälften (44,4%) av alla respondenter kände sig stigmatiserade på grund av migrän. Anledningarna till denna stigma och de föreslagna lösningar på hur man kan minska/hantera stigman var ganska lika. Okunnigheten hos andra var den mest rapporterade orsaken till deras migränstigma, och ökad kännedom och korrekt information om migrän var det mest rapporterade sättet att minska stigmatiseringen. Många av de tillfrågade hade, på grund av sin migrän, utsatts för nedvärdering på jobbet, från familj och vänner och från hälsovårdpersonal. Utsättning för nedvärdering från hälsovårdpersonal rapporterades ha hänt ofta av 11,5% och ibland av 34,7% av alla respondenter. Av alla respondenter oroade sig 55,6% ofta och 29,8% oroade sig ibland för början av nästa migränattack. Majoriteten av respondenterna hade svår migränrelaterad funktionsnedsättning baserat på deras MIDAS-grad. Många andra aspekter av bördan rapporterades också, inklusive stigma, vilket rapporterades av nästan hälften av respondenterna. Ytterligare och framtida studier måste genomföras för att få en ännu bättre förståelse för migränens börda och stigma hos vuxna finländska migränpatienter. Detta inkluderar ytterligare och mer invecklade kvantitativa och kvalitativa analyser av datan från denna studie, liksom studier med nya perspektiv baserade på resultaten av denna studie.
  • Parkkinen, Ilmari (Helsingfors universitet, 2018)
    MicroRNAs are ~22 nucleotide long RNA strands which regulate gene expression by binding to the 3’UTRs of messenger RNAs. MicroRNAs are predicted to regulate about a half of all protein-coding genes in the human genome thus affecting many cellular processes. One crucial part of microRNA biogenesis is the cleaving of pre-miRNA strands into mature microRNAs by the type III RNase enzyme, Dicer. Dicer has been shown to be downregulated due to aging and in many disease states. Particularly central nervous system disorders are linked to dysregulated microRNA processing. According to the latest studies, Dicer is crucial to the survival of dopaminergic neurons and conditional Dicer knockout mice show severe nigrostriatal dopaminergic cell loss, which is a hallmark of Parkinson’s disease. By activating Dicer with a small-molecule drug, enoxacin, the survival of dopaminergic cells exposed to stress is significantly improved. However, enoxacin, which is a fluoroquinolone antibiotic, activates Dicer only at high concentrations (10-100 μM) and is polypharmacological, which may cause detrimental side effects. Therefore, enoxacin is not a suitable drug candidate for Dicer deficiencies and better Dicer-activating drug candidates are needed. The aim of this work was to develop a cell-based fluorescent assay to screen for Dicer-activating compounds. Assays which measure Dicer activity have already been developed, but they have some pitfalls which don’t make them optimal to use for high-throughput screening of Dicer-activating compounds. Some are cell-free enzyme-based assays and thus neglect Dicer in its native context. The RNA to be processed by Dicer does not represent a common mammalian RNA type. Most assays do not have internal normalizing factors, such as a second reporter protein to account for e.g. cell death, or the analysis method is not feasible for high-throughput screening data. Considering these disadvantages, the study started by designing a reporter plasmid in silico. The plasmid expresses two fluorescent proteins, mCherry (red) and EGFP (green), and a mCherry transcripttargeting siRNA implemented into a pre-miR155 backbone which is processed by Dicer. Thus, measuring the ratios of red and green fluorescence intensities will give an indication on Dicer activity. The plasmid also has additional regulatory elements for stabilizing expression levels. The plasmid was then produced by molecular cloning methods and its functionality was tested with Dicer-modulating compounds. The assay was optimised by testing it in different cell lines and varying assay parameters, and stable cell lines were created to make large-scale screening more convenient. Finally, a small-scale screen was done with ten pharmacologically active compounds. Transiently transfected, in Chinese hamster ovarian cells, mCherry silencing was too efficient for reliable detection of improvement in silencing efficiency due to floor effect. With an inducible, Tet-On, system in FLP-IN 293 T-Rex cells, the expression could be controlled by administering doxycycline and the improvement in silencing was quantifiable. The assay seemed to be functional after 72 hours and 120 hours of incubation using enoxacin (100 μM) as a positive control. However, the screening found no compounds to significantly reduce mCherry/EGFP fluorescence ratio and, additionally, the effect of enoxacin was abolished. Therefore, a more thorough analysis on the effects of enoxacin was done and, although statistically significant, enoxacin was only marginally effective in reducing mCherry/EGFP fluorescence ratio after 72 hours of treatment. It should be noted from the small-scale screening that metformin and BDNF, compounds previously shown to elevate Dicer levels, showed similar effects to enoxacin. The quality of the assay in terms of high-throughput screening was determined by calculating Zfactors and coefficients of variations for the experiments, which showed that the variability of the assay was acceptable, but the differences between controls was not large enough for reliable screening. In conclusion, the effects of metformin and BDNF should be further studied and regarding the assay, more optimisation is needed for large-scale, high-throughput, screening to be done with minimal resources.
  • Sjöberg, Madeleine (Helsingfors universitet, 2018)
    Cancer afflicts an ever-growing number of people globally each year. In part due to a complex pathophysiology where much is still unknown, the need for new cancer treatments has been persistent, fuelled further by the issue of treatment resistance. An emerging field holding much promise in oncology is immunotherapy, a subgroup of which is oncolytic virus treatments. These treatments utilize the inherent or acquired ability of certain viruses to selectively replicate in tumor cells to fight cancer. One of these viruses is the adenovirus. With these viruses it is possible to modulate the immune response e.g. through the expression of certain genes. The thesis focuses on genetically arming an oncolytic adenovirus in an effort to enhance treatment efficacy. The transgene of choice is the CD40 ligand (CD40L), a costimulatory molecule capable of aiding in the development of systemic antitumor immunity. Adenoviruses have previously been designed expressing the CD40L, however, a novel aspect was introduced with the design and incorporation of a soluble a form of the protein. The main aim of the study was the construction of four functional oncolytic adenoviruses, encoded with either the human or mouse variants of the two CD40L proteins (full-length and soluble). Successful completion required protocols for the cloning, bacterial colony screening, and primary virus production to be established. Insertion of the CD40L transgenes into the E3-gp19k region of the chosen Ad5Δ24 backbone was first attempted with the traditional approach of homologous recombination. The method that ultimately proved successful was a one-step Gibson Assembly® reaction. Screening the bacterial colonies with colony polymerase chain reaction, the potential CD40L positive clones underwent restriction analysis to affirm the presence of the transgene in the viral genome, as well as the retainment of critical elements. Two out of three recombined plasmids carrying the full-length CD40L proceeded to transfection and virus propagation in A549 cells, after which the presence of the adenovirus and CD40L expression was confirmed with immunostaining. Finally, a protocol was successfully established by the construction of one of the intended four viruses. The protocol entails all the main steps from cloning until primary virus production, additionally offering the option of applying it to the genetic arming of the Ad5Δ24 with other transgenes of interest. In terms of future perspectives for the project, following construction of the remaining viruses, the intentions are to validate transgene expression and functionality for all constructs, as well as compare the immunogenicity between the full-length and soluble CD40L. In the event of promising results, the project will hopefully proceed to in vivo studies.
  • Honkasalo, Oona (Helsingfors universitet, 2018)
    Cancer immunotherapies aim to target the immune defence mechanisms of the body specifically and efficiently against the tumour tissue. Cancer vaccines and oncolytic viruses are forms of active immunotherapies, which require patients having a properly functioning immune system. The vaccines are based on the administration of tumour antigens into the body to which the immune system reacts. However, often the response is not robust enough. The oncolytic viruses in turn kill the cancer cells which causes the release of antigens from the tumour tissue. Viruses usually elicit a strong immune response but sometimes it is targeted too much against the virus instead of the tumour. Oncolytic vaccine is a composition of an oncolytic virus and a cancer vaccine. Tumour antigens can be coded to the genome of the virus therefore, when the virus invades tumour cells they start to produce the antigens. Eventually the cancer cells are also destroyed due to viral replication. The antigens can be tumour-associated that is, they are expressed in healthy tissues too. Their usage is not always efficient which is why an interest towards utilizing tumour-specific antigens has been increased. Considering the expression of antigens, tumour tissue is very heterogenous and distinctive between patients. Hence, utilizing mutated patient unique neoantigens would enable the development of personalized tumour-specific oncolytic vaccines. Genetic modification of viruses is complicated thus, an easier way to insert the neoantigens to the virus has been invented. The developed oncolytic vaccine platform is called PeptiENV, and it is designed to use with enveloped viruses. The idea is to fuse tumour-specific antigens onto the envelope of the virus and eliminate the need of gene insertion. The aim of this study is to investigate in vivo the efficacy of PeptiENV in preventing tumour growth and eliciting a tumour-specific immune response. An object is also to observe survival times of the treated animals. Furthermore, the preservation of infectivity is studied in vitro. The research was executed with two potential oncolytic viruses, vaccinia virus (VACV) and herpes simplex virus type 1 (HSV-1). The PeptiENV complex was formed by using an artificial tumour antigen, ovalbumin epitope SIINFEKL, which was attached to the viral envelope with cell penetrating peptide (CPP) or cholesterol anchor. The preservation of infectivity was examined by measuring cell viability of PeptiENV infected cells. Animal experiments instead were performed with a mouse melanoma model created with B16-OVA cells, which express ovalbumin and therefore the antigen epitope SIINFEKL. PeptiENV was compared to control treatments which were virus, SIINFEKL peptide and complexation medium only. Treatments were administered as intratumoural injections. Tumour growth was followed by measuring the size of implanted tumours every other day. With flow cytometry, tumour-specific immune response was assessed by acquiring the relative amount of SIINFEKL-specific CD8+ T cells in the tumour tissue. Euthanizing dates were registered in order to observe the survival of the mice. According to the in vitro results, conjugation of peptides to the virus does not affect infectivity. In addition, the in vivo studies show that PeptiENV VACV CPP prevents tumour growth the most. Difference in tumour growth between PeptiENV VACV CPP and control treatments is significant. Mice injected with the same treatment also lived considerably longer than mice injected with virus, peptide or medium only. Also, PeptiENV HSV-1 hinders tumour growth distinctly more than virus only and slightly more than SIINFEKL only, but unfortunately it did not have an evident impact on the survival time. In both experiments, the PeptiENV treatment elicits the largest proportional amount of SIINFEKL-specific CD8+ T cells. In other words, PeptiENV engenders a tumour-specific immune response. In the PeptiENV VACV study the difference to control treatments is clearer than in the PeptiENV HSV-1 study. At present, the PeptiENV platforms performs better with VACV than HSV-1. With further investigations however, the results can be verified and improved. All in all, the results are encouraging. The PeptiENV platform shows great promise for being a part of personalized cancer immunotherapy developments in the future.
  • Tepsell, Juhani (Helsingfors universitet, 2018)
    During and after myocardial infarction, millions to a billion cells die off. Scar tissue formed by fibroblasts replaces the injured myocardium during recovery. While the newly formed tissue is durable and prevents rupture of the heart, it doesn´t contribute to pump function. Depending on the extent of cardiomyocyte loss, the remaining functional myocardium get strained. Adult mammalian heart has inadequate capacity to regenerate after such injury. In case of sustained substantial increase in workload, the compensatory mechanisms turn into pathological processes including excessive fibrosis and myocyte apoptosis. The progressive decline of hearts contractile function results in heart failure (HF). Current drug treatments for managing HF aim to prevent progression of the disease and relieve symptoms. ACE inhibitors, beta blockers and diuretics are effective along with healthy lifestyle. No practical treatments are available to restore cardiac function yet. Human myocardium normally regenerates, but only 1% or less of myocytes get replaced yearly. Heart’s resident stem/progenitor cells (CPCs) likely play a role in the turnover. The aim of this study was to develop a screening method to identify small molecules that possibly promote differentiation of cardiac progenitor cells to cardiomyocytes. Cell population differentiated from mouse embryonic stem cells (mESCs) was used as a model for CPCs. Directed differentiation protocol of mESCs used here promotes commitment to cells of cardiac mesoderm, part of which will further differentiate to cardiac progenitors. The resulting population at day 6 is heterogenous but many of these cells are progenitors that turn into cardiomyocytes (CMs) by day 8. 10 000 cells per well are plated on 384 well plates at day 5. Test compounds are added at day 6 and removed day 8 for effect in progenitors and day 7-9 for effect in early cardiomyocytes. 0,1% DMSO is used as vehicle and Wnt pathway inhibitor XAV939 as positive control. The effects are quantified with plate reader on day 9. E14 derived mESC reporter line was used. Myl2v-eGFP + SMyHC3-RFP double reporter line allows the specific identification of ventricular CMs with green fluorescence and atrial CMs with red fluorescence. Plate reader measures the total fluorescence of the wells at 485/520nm on day 9, which is used as a readout for ventricular CMs. The fluorescence intensity depends on the amount of GFP+ cells but also on the level of Myl2v expression. Atrial CMs could be quantified similarly but the population doesn´t contain enough RFP+ cells. The assay was shown to reliably point out ‘hits’ that have a strong effect. Any compounds that only produce a moderate effect could be a false negative, however. The effect on cardiac progenitors could likely be increased by simply adding the compounds earlier on day 5. Variability of key reagents causes the main technical troubles through unpredictably affecting cytokine concentrations which decreases the amount of cardiac progenitors. Partially similar screening assays are being used by the big pharma where they cryopreserve progenitors in bulk for later use, thus simplifying and speeding up their method. Same approach could be adopted.
  • Vesterinen, Johanna (Helsingfors universitet, 2012)
    Pyrazoles are five-membered nitrogen containing heterocycles, whose derivatives can be widely used in medicinal chemistry. One of the most common ways to produce pyrazoles is 1,3-dipolar cycloaddition, where the dipole containing heteroatom is reacting with a dipolarophile, and forming a cycloadduct. Among others, mesoionic sydnones have been used as dipoles in 1,3-dipolar cycloadditions of pyrazoles. During the last decades solid phase methods, where either a dipole or dipolariphile is being temporarily bound to solid support, have been exploited in 1,3-dipolar synthesis. With the aid of solid phase methods, the synthesis can be controlled chemically by protecting groups that react easily. Also the isolation and purification can be made easier by using these techniques. The 1,3-dipolar solid phase methods can be combined with microwave techniques, to make the synthesis shorter and more effective. The goal of this work was to synthesize new N-unsubstituted pyrazoles, starting from sydnones bound to solid support and alkynes, with use of 1,3-dipolar cycloaddition reaction and to purify and analyze prepared compounds. The aim was also to develop a new 1,3-dipolar solid phase method so, that the end products could be cleaved easily in a traceless manner, and that there would be minor need for purification. There was also an aim to make the last step of the synthesis faster and easier with microwave reactor, and by using this method to standardize the conditions used in the cycloaddition-dehydration reaction step. The AMEBA-resin was chosen to be the solid support in this synthesis. Its traceless cleavage by trifluoroaceitic acid made possible not only the formation of the desired structure, but also effortless purification of the end product. The amino group of pyrazole was protected by the solid support during the N-nitrosation, so that after the traceless cleavage of the resin, N-unsubstituted pyrazoles were obtained. By changing the amino acids used in this synthesis, it was possible to alter the structure of the synthesized pyrazoles, and to create four structurally new pyrazoles. Microwave method used during the last step of synthesis for heating shortened the reaction step significantly, and also the yields of end products were better compared to conventional heating. During this work a functioning and developable 1,3-dipolar solid phase method was created, that can be utilized to synthesize pyrazoles and other compound of similar nature also the in future.
  • Hynynen, Marko (Helsingfors universitet, 2017)
    Smoking poses a significant threat to public health. Major public health benefits could be attained if we could increase the incidence of smoking cessation on the national level. However, smoking cessation is often difficult, therefore different medical means have been devised to make it easier for people to quit. One of these medical means is nicotine replacement therapy(NRT). Since NRT entered the market it has been contested which kind of role counselling should have when people purchase NRT. NRT was deregulated In Finland in 2006. Prior to this thesis there has not been carried out studies that would have tried to investigate where the retailer shops are located after deregulation. The aim of this thesis is to investigate how the deregulation affected to the distribution of NRT sales in different NRT-outlets and pharmacies in 2006-2015. It is also aim to investigate if the new outlets have tangibly increased the coverage of NRT-outlets network in diverse municipalities, e.g.in more sparsely populated rural municipalities. Finnish Medical Agency‘s (FIMEA) data about NRT-outlets between the years of 2006-2015 was used as source data for this thesis. This data was analysed by comparing the number of NRToutlets and pharmacies in diverse municipalities. The municipalities have been classified regarding the area and population of diverse municipalities, among other parameters. The PDF sheet of FIMEA’s NRT-outlet data was converted into an Excel file, and using that as a basis different diagrams were made. The number of NRT-outlets grew steadily until during the year 2011 there was a slightly bigger increase in the number of NRT-outlets as NRT became available in restaurants too. Afterwards the growth of NRT-outlets diminished. It can be seen from the results that NRT-outlets, more than pharmacies, are concentrated to the crowded municipalities. On the other hand, there has hardly been an increase in the coverage of NRT-outlet network in more sparsely populated rural municipalities since deregulation took place. However, many of the municipalities that have a pharmacy and one or a few other NRT-outlets are located in Southern Finland surrounding bigger city municipalities or in Central Finland.
  • Volotinen, Katariina (Helsingfors universitet, 2012)
    Kolinergiset α6*-nikotiinireseptorit ovat kiinnostavia, koska ne liittyvät mahdollisesti Parkinsonin tautiin ja nikotiiniriippuvuuteen. Ionikanavina toimivat nikotiinireseptorit ovat muodostuneet viidestä alayksiköstä, jotka esiintyvät erilaisina yhdistelminä. α6-alayksikköä sisältävät nikotiinireseptorit sijaitsevat presynaptisesti ja säätelevät dopamiinin vapautumista dopaminergisessä hermopäätteessä. α6*-nikotiinireseptorit ovat keskittyneet vain tietyille aivoalueille ja niitä esiintyy runsaasti dopaminergisissä hermosoluissa. α6*-nikotiinireseptoreita on erityisesti mesolimbisen ja nigrostriataalisen hermoradan dopaminergisissä hermosoluissa. Lisäksi niitä on paljon näkemiseen liittyvillä aivoalueilla. Nikotiini toimii asetyylikoliinin tavoin aktivoimalla α6*-nikotiinireseptoreita, mikä johtaa dopamiinin vapautumiseen hermopäätteessä. α6*-nikotiinireseptoreiden sijainnin, määrän ja toiminnan tutkimisessa on käytetty apuna muun muassa niille selektiivisiä antagonisteja, saalistavista merietanoista peräisin olevia α-konotoksiineja, erityisesti α-konotoksiini MII:ta. Nigrostriataalisella hermoradalla, joka ulottuu substantia nigrasta striatumiin, α6*-nikotiinireseptorit voivat vaikuttaa liikkeen säätelyyn. Nikotiini vapauttaa dopamiinia nigrostriataalisen hermoradan päätepisteessä, striatumissa, mikä voi lisätä liikeaktiivisuutta. Nikotiinilla on havaittu olevan hyödyllisiä vaikutuksia Parkinsonin taudin eläinmalleissa, mutta Parkinsonin tautipotilailla nikotiinihoidosta saadut tutkimustulokset ovat ristiriitaisia ja puutteellisia. α6*-nikotiinireseptoreille voitaisiin kehittää selektiivisiä agonisteja, joiden avulla lääkehoito voitaisiin kohdentaa paremmin ja vältyttäisiin mahdollisilta haittavaikutuksilta. Tupakanvieroitukseen tarvittaisiin lisää uusia selektiivisiä lääkehoitoja, joilla olisi hyvä hoitomyöntyvyys ja mahdollisimman vähän haittavaikutuksia. Mesolimbinen hermorata, joka ulottuu ventraaliselta tegmentaalialueelta nucleus accumbensiin, liittyy riippuvuuden syntyyn. Nikotiinin vaikutukset välittyvät VTA:n kautta nucleus accumbensiin, jossa vapautuu dopamiinia. Osa nikotiinin vaikutuksista välittyy myös presynaptisten α6*-nikotiinireseptorien kautta. Selektiivisistä α6*-nikotiinireseptoreiden antagonisteista voisi olla hyötyä nikotiiniriippuvuuden hoidossa, sillä niiden vaikutus vastaisi osittaisagonistin vaikutusta. α-konotoksiini PIA:n vaikutuksia nikotiinin aiheuttamaan dopamiinin vapautumiseen tutkittiin in vivo mikrodialyysimenetelmällä. α-konotoksiini PIA saalistavasta merietanasta eristetty selektiivinen α6*-nikotiinireseptoreiden antagonisti. Tutkimuksessa käytettiin vapaana liikkuvia urospuoleisia Wistar-rottia. Tutkimuksen kohteena olevat aivoalueet olivat striatum ja nucleus accumbens. Rotille asennettiin anestesiassa ohjauskanyyli joko striatumiin tai nucleus accum-bensiin stereotaktisen laitteen avulla. Mikrodialyysikokeessa koetinten tasapainotuksen jälkeen kerättiin perustason näytteet ja pistettiin saliini tai nomifensiini tai vaihdettiin Ringer-ruiskun tilalle α-konotoksiini PIA-ruisku. Puolen tunnin päästä pistettiin saliini tai nikotiini ja vaihdettiin Ringer-ruisku takaisin. Näytteitä kerättiin 15 minuutin välein yhteensä 5,5 tuntia. Lopuksi aivot otettiin talteen ja niistä tehtyjen aivoleikkeiden avulla tarkastettiin koetinten paikat. Mikrodialyysinäytteistä määritettiin HPLC-menetelmällä dopamiinin ja sen metaboliittien DO-PAC:n ja homovaniliinihapon sekä koejärjestelyssä oletettavasti muuttumattomana pysyvän 5-HIAA:n pitoisuudet. Koejärjestelyssä päätettiin käyttää tutkittavien aivoalueiden dopamiinipitoisuuden nostamiseen nomifensiinia, joka estää dopamiinin takaisinottoa hermopäätteissä. Käsittelyryhminä olivat saliini-saliini (n=striatum ja nucleus accumbens, 8+7), saliini-nomifensiini (n=8+4), saliini-nikotiini (n=3+4), nomifensiini-nikotiini (n=10+13) ja nomifensiini-nikotiini-α-konotoksiini PIA (n=8+5). Rottia jouduttiin hylkäämään eri syistä joko ennen mikrodialyysia, mikrodialyysin aikana tai sen jälkeen. Rottia hylättiin yhteensä 70 kpl. Tilastollisessa analyysissä tutkittujen käsittelyiden tai aivoalueiden välille ei saatu merkitseviä eroja, koska eläinten välinen hajonta oli liian suurta. Silmämääräisesti nomifensiini-nikotiinikäsittely nosti striatumin ja nucleus accumbensin dopamiinipitoisuuksia. α-konotoksiini PIA näytti estävän dopamiinin vapautumista striatumissa ja nucleus accumbensissa, mutta erot nomifensiini-nikotiinikäsittelyyn eivät olleet tilastollisesti merkitseviä. AUC-arvolla mitattuna α-konotoksiini PIA esti dopamiinin vapautumista striatumissa 39,6 % ja nucleus accumbensissa 31,3 %. Aivoalueiden välillä ei ollut tilastollisesti merkitseviä eroja.
  • Hautaniemi, Mikaela (Helsingfors universitet, 2012)
    In pharmaceuticals amorphous state can be obtained either intentionally or unintentionally. Intentional production is used, for example, to improve the dissolution of poorly soluble compounds, to stabilize the structure of proteins, or to improve the mechanical properties of excipients (e.g., lactose). Unintentional introduction of amorphous phases can result from general manufacturing procedures of pharmaceuticals, such as coating, granulation, drying, milling, and compression. The presence of amorphous regions, even in small quantities, can exhibit a significant influence on the physical and chemical stability of pharmaceutical products. Molecular mobility in formulation with amorphous content is believed to be the key factor of their stability. Therefore, evaluating of molecular mobility is an important step in pharmaceutical product development. The aim of this study was to estimate molecular motions in amorphous disaccharides using calorimetric approach at temperatures below the glass transition temperature (Tg), where relaxation process is very slow as compared to the time of experiment. When temperature is low enough, the initial relaxation time parameter (τi) can be used as an estimate for relaxation process on the timescale of pharmaceutical product shelf life. The results of the present study revealed similar trend in stability of amorphous forms for the disaccharides (sucrose experiencing the fastest structural relaxation), which can be assumed on the basis of Tg alone, where higher Tg would result in more stable glassy state (Tg of sucrose is the lowest). Storage temperature of Tg - 55oC or lower would suffice for amorphous trehalose, melibiose and cellobiose to achieve at least 2 year's relaxation time, while for sucrose the temperature is Tg - 70oC. Fragility has been used as a helpful mean for classifying amorphous materials. All the compounds can be classified as fragile. Fragility ranking in the present study contains some degree of uncertainty, while 3 different approaches revealed somewhat different results for ranking the disaccharides. The variation in the results can be attributed to the overall sensitivity of DSC. The method described in the present study is quite difficult to apply without supportive information from other techniques. The results, obtained with the method, are very dependent on the slope in plotting ln q vs. 1/Tg, and even small fluctuations in the estimation can lead to different fragility values and consequently to different relaxation times. However, the final results reveal values for relaxation times well below Tg, which are in reasonable agreement with modern theoretical understanding of glassy state dynamics.
  • Viljemaa, Kati (Helsingfors universitet, 2017)
    The economic burden of adverse events (AEs) is substantial and in direct relation to current increasing drug utilisation. According to previous research, the annual cost of AEs in the U.S. may be as high as 22.9 billion euros. In Europe AEs are considered to contribute to 3.6 percent of hospital admissions, have an impact on 10 percent of inpatients during their hospital admission and are responsible for less than 0.5 percent of inpatient deaths. AEs thus clearly constitute a major clinical issue. Fluoroquinolones have been in clinical use since the 1980s and are globally among the most consumed antimicrobials. Fluoroquinolones are generally well tolerated antimicrobials. The most common AEs are mild and reversible, such as diarrhea, nausea and headaches. Nevertheless, fluoroquinolones are also associated with more serious AEs, including Clostridium difficile associated diarrhea (CDAD), rate-corrected electrocardiographic prolonged QT interval, tendinitis and tendon rupture, dysglycemia, hepatic toxicity, phototoxicity, acute renal failure and serious AEs involving the central nervous system, such as seizures. Health service use and costs specifically associated with fluoroquinolone-related AEs have not been evaluated previously. The theory section of this Marter's thesis considers adverse events and fluoroquinolones. The main principles of conducting a systematic review are also discussed. The empirical section is a systematic review. The aim of this study is to identify health care use and costs associated with ciprofloxacin, levofloxacin, moxifloxacin, norfloxacin and ofloxacin -related AEs. A literature search covering Medline, SCOPUS, Cinahl, Web of Science and Cochrane Library was performed in April 2017. Two independent reviewers systematically extracted the data and assessed the quality of the included studies. All costs were converted to 2016 euro in order to improve comparability. Of the 5,687 references found in the literature search, 19 observational studies, of which 5 were case-controlled, fulfilled the inclusion criteria. Hospitalization was an AE-related health care use outcome in 17 studies. Length of stay associated with AEs varied between <5 - 45 days. The estimated cost of an AE episode ranged between 140 and 18,252 €. CDAD was associated with the longest stays in hospital. However, a mere 10 studies reported AE-related length of stays and only 5 evaluated costs associated with AEs. Although rare, in particular serious fluoroquinolone-related AEs can have substantial economic implications, in addition to imposing potentially devastating health complications for patients. Further measures are required to prevent and reduce health service use and costs associated with fluoroquinolone-related AEs. Equally, better-quality reporting and additional published data on health service use and costs associated with AEs are essential. The strengths of this study are a comprehensive and systematic literature search and transparency of methodologies and reporting. The main weakness is the generalizability of the results.
  • Kallio, Rosanna (Helsingfors universitet, 2017)
    A large part of patient safety incidents in health care is related to medicines and medication treatment. Medication safety is an important part of patient safety. In particular, transitions of care endangers continuity of care and patient safety. A poor flow of information between health care units increases a risk of medication errors. An accurate and up-to-date medication list can improve transfer of correct medical information with the patient. In addition to the medication list other organization-level defences are important to patient and medication safety. This study is part of a larger regional development project concerning reformation of service delivery in health and social care. One of the purposes of the project is to integrate health and social services of municipalities in central Uusimaa (Hyvinkää, Mäntsälä, Pornainen, Järvenpää, Nurmijärvi and Tuusula). The aim of this study was to investigate what kinds of medication lists are available in these municipalities and to design a medication list maintained by the patient which is regionally shared. The second aim of the study was to identify defences used by the municipal healthcare systems and to develop a medication management process model for the municipalities. The material consisted of medication lists from the municipalities participating in the study. Twelve (12) lists were selected for analysis. The medication list of Lääkekortti.fi was also included in the analysis (N=13). Data of the defences was collected by a questionnaire. In Hyvinkää the material was collected from primary health care and Hyvinkää hospital. Both the medication list and the defence data were systematically reviewed and collected in a Microsoft Excel table. The regional medication list for patients was developed based on the medication list data and the existing literature. The process model for medical treatment was developed based on the defence data. Almost every medication list contained space for personal data (n=11) and basic medication information, such as the name of the drug (n=12), strength (n=12) and dosage (n=13). Regular medication was usually separated from as-needed medication (n=8). Only two of the lists had a column for periodic medication. There were only two lists with columns for over-the-counter medicines and herbal products. Less than half of the lists (n=5) had space for indication. According to the survey, most of the defences listed in the questionnaire were used in all or almost all municipalities. According to the responses, medication lists, for example, are regularly reviewed. Some gaps in defences also emerged. Only in two municipalities high-alert medications were identified. Also only in two municipalities attention was paid to the storage of look-alike medicines. Written instructions for managing medication errors was available only in two municipalities. There are many different medication lists available in health care. A regionally shared medication list maintained by the patient can improve the transfer of the up-to-date medical information with the patient. The list must be comprehensive and easy to use. Both the patient and the healthcare professionals should be responsible for maintaining the accurate medication list. Most of the defences asked in the questionnaire have been taken into account in pharmacotherapy plans. In practice, however, the implementation of defences is unclear. For example patients' medication lists often contain errors even though lists should be reviewed regularly. There should be exact instructions as well as clearly defined roles and responsibilities for medication reconciliation. More attention should also be paid to the use of high-alert and look-alike medicines. Since errors occur despite the defenses, every health care unit should have written instructions in case of medication errors.
  • Huovila, Tiina (Helsingfors universitet, 2017)
    Autophagy is a pathway for cells to degrade intracellular components that are no longer needed or are detrimental for the cells. It is essential for cell homeostasis and survival and has been related to various diseases and pathophysiology. Autophagy is a complex process and there are still several unclear und unknown aspects to it. Regulation of autophagy is essential to prevent unwanted and escess activation, and several pathways and molecules, both stimulatory and inhibitory, are included. Different signaling pathways are sensitive to a variety of environmental clues. Two main autophagy pathways are mTOR-dependent pathway and mTOR-independent pathway. Induction of autophagy in the latter pathway is dependent on the interaction of Bcl-2 and Beclin 1. Prolyl oligopeptidase (PREP) is a peptidase enzyme that has several substrates. PREP-inhibition by KYP-2047 can reduce aggregation of α-synuclein in two ways: by increasing rate of autophagy and by decreasing dimerization. The aim of this study was to find out how PREP affects the interaction between Bcl-2 and Beclin 1 and how this affects autophagy. Based on previous studies, PREP-inhibition seems to increase the amount of Beclin 1 and to affect the phosphorylation of Bcl-2 and Beclin 1, leading to dissociation of the complex. Hypothesis was to see differences in colocalization of Bcl-2 and Beclin 1 in cells treated with different PREP-modifications and for PREP-inhibition to decrease the colocalization. Human embryonic kidney cells 293 (HEK-293) and hPREP knockout cell line created from them by using CRISPR/Cas9-silencing were used in the experiments. Two experiments were performed on regular HEK-cells: inhibitor experiment with KYP-2047 (1 or 10 µM) and overexpression experiment (transfection with either active or inactive hPREP plasmid). After immunofluorescence staining, cells were analysed with confocal microscope and colocation analysis of Bcl-2 and Beclin 1 was performed. The intensity of Beclin 1 in the nuclei was stronger than in other parts of the cell in all samples, which could indicate a stronger activity of its nuclear tasks compared to autophagy. However, the antibody used for immunofluorescence has most likely caused this staining pattern. Based on previous knowledge, it was expected to see differences in colocalization of Bcl-2 and Beclin 1 in cells treated with different PREP-modifications. However, there were no significant differences in colocalization of Beclin 1 and Bcl-2 in any of the experiments but it was nearly 100 percent in all treatments. Since rate of autophagy in cells was not detected, it is impossible to determine, if there were changes in autophagy that were not reflected as changes in colocalization of these two proteins. It is possible that even a small change in colocalization can affect the rate of autophagy or there might be subpopulations where the interaction is interrupted and these changes are so small that they are not detectable with the methods used in this experiment. Both Bcl-2 and Beclin 1 also have functions not related to autophagy, which could be one reason behind the results gained in this study.
  • Pihlaja, Tea (Helsingfors universitet, 2017)
    Cytochrome P450 (CYP) enzymes are important catalysers in the first phase of drug metabolism. Roughly two thirds of drugs are oxidized via CYP enzymes, which enable the further modification of drugs, and their excretion. In this thesis, human liver microsomes containing the main hepatic CYP enzymes were immobilized on thiol-ene based micropillar arrays and their stability was evaluated using a CYP2C9 isoenzyme specific luminescent substrate, Luciferin-H. The aim of the study was to develop microfluidic immobilized enzyme reactors (IMERs) for studying enzyme kinetics and drug-drug interactions. For this purpose, the instability issues associated with previously reported CYP-IMERs were carefully addressed. The CYP immobilization protocol used was based on a protocol previously developed in the context of other research projects and relied on biotinylation of human liver microsomes (HLM) with help of fusogenic liposomes. The biotinylated HLMs were then attached to the streptavidin-modified thiol-ene surfaces. The CYP activity was determined by utilizing microfluidics under continuous flow conditions (typically 5 μL/min) in the presence of NADPH. The luminescent metabolite formed by the CYP2C9 enzymes was quantified with a commercial well-plate reader from fractions collected at the microreactor outlet. Half-life was used to compare the differences between enzyme stabilities reached via different immobilization conditions. The effects of flow rate and reaction temperature on the stability of the CYP-IMERs was evaluated together with addition of antioxidative agents and reactive oxygen species (ROS) scavengers. Different functionalization steps as well as storage time and conditions were studied. With Luciferin-H as the model substrate of CYP2C9, the CYP-IMERs showed higher activity and stability at room temperature than at +37 °C. The peak activity could be increased via optimization of the immobilization protocol, though long-term storage diminished the peak activity. The activity of the IMERs typically attenuated within 1-2 hours with little or no improvement achieved via optimization of the immobilization or operation conditions. Only upon addition of the ROS scavengers, the peak activity and stability of the CYP-IMERs could be slightly improved. After functionalization, the IMERs maintained their activity until the time of use when stored in +4 °C for up to 2 weeks, but re-use of IMERs was not possible.
  • Jakola, Janne (Helsingfors universitet, 2017)
    The incidence of wet age-related macular degeneration (AMD) is increasing with ageing. AMD leads to blindness if it is not treated properly. Common treatment is to administrate intravitreal growth factor inhibitors. An ageing population increases the number of patients which overloads the public health services and expands costs. Traditionally, injections have been administered by physicians but because of the limited recourses nurses have been trained to administer injections. In addition, injections can be administered as physician's clinical extra work to alleviate the queue and as an outsourcing service from the private sector. As the resources of the public health service are limited, it is important to evaluate used methods reliably. The target of this research was to investigate the administration costs of intravitreal injections which are administered by physicians, nurses, a physician or a nurse working extra to alleviate the queue or by a private sector. The used method was cost analysis because the effectiveness of the care is the same regardless of the administer of the injection. The source of costs was Ecomed database of HUS, the data of cost accounting and catalogue of billing from the outsourcing service. The costs were examined in perspective of the producer of the service. Based on the cost analysis, the administration costs per injection are following: administered by physicians 51,39 €, nurse-administered 51,19 €, administered by a physician to alleviate the queue 100,46 €, nurse-administered to alleviate the queue 72,87 €, administered by a physician in outsourcing service 276,19 and nurse-administered in outsourcing service 269,85 €. The annual total costs of the producer of the service were 4 563 726 €. By increasing the number of injections administered by a nurse of HUS the need for an outsourcing service can be decreased which may decrease the annual total costs by two million euro. It is important to find cost-effective solutions because the number of patients are increasing. Based on this research it is more profitable to increase the number of injections administered by a nurse of HUS than to train more nurses to administrate injections to alleviate the queue or to work in outsourcing services. The result of this research can be adapted in planning of the public health services.
  • Tilli, Irene (Helsingfors universitet, 2017)
    Melanoma is the most severe case of skin cancer and there is no curative treatment if it has progressed. Despite the recent advances in drug therapy tens of thousands of patients die of melanoma annually. There is still need for new antimelanoma drugs for which marine compounds are a potential source. Halogens are common elements in drug molecules as they enhance their molecular properties. So far most of the halogenated drugs contain fluorine and/or chlorine but the role of bromine and iodine is probably growing in the future due to halogen bonding. Bromotyrosines are originally isolated from Verongiida-order sponges but whether they are truly of bacterial origin is under controversy. All bromotyrosine compounds consist of brominated tyrosine and/or tyramine residues or their derivatives. Purpurealidin I is one of the newest bromotyrosine derivatives extracted from Pseudoceratina purpurea and it has shown activity against melanoma. In this study eight new purpurealidin I derivatives were synthesized following a successful route previously designed. All synthesized derivatives contained the original N-oxime structure which's stereochemistry was determined to be E by X-ray crystallography. Cytotoxicity against A375 melanoma cells was determined for seven compounds synthesized here and for 15 compounds synthesized previously. All seven compounds and one previously synthesized purpurealidin I analog were active with CC50 values between 4,7 and 22,1 µM. The previously synthesized bromotyrosine derivative intermediates and aerophobin-1 analogs were not active. The selectivity of the active compounds was calculated by determining their CC50 value against Hs27 fibroblast cells. None of the compounds showed remarkable selectivity the most selective 2-pyridin containing derivative having four times better selectivity against melanoma. The tyrosine part and N-oxime seem to be important parts to preserve while the tyramine part can be modified more freely and maintain the activity. Still more derivatives need to be synthesized and tested to confirm these observations. More data is also needed considering the selectivity of the compounds.
  • Somersalo, Petter (Helsingfors universitet, 2017)
    Cells release different types of phospholipid bilayer-limited vesicles into the extracellular space. These are commonly referred to as extracellular vesicles (EVs). Exosomes (EXOs), ca 50-100 nm in diameter and microvesicles (MVs), ca 100-1000 nm in diameter, having different intracellular origin, are the two main subpopulations of EVs. EVs have been demonstrated to carry a range of proteins and nucleic acids subsequently delivered to recipient cells, making them attractive as drug delivery vehicles. Several mechanisms for the cellular uptake of EVs have been established. When a nanoparticle is introduced into blood plasma, plasma proteins are adsorbed to its surface, forming a protein corona. The formation of the corona is a dynamic process, governed by individual protein concentrations as well as their respective affinities for the surface. Proteins of the corona interact with surrounding cells, thus being able to influence the cellular uptake of the nanoparticle. In the current study, the uptake of PC-3-derived EVs into PC-3 cells was investigated. Moreover, the impact of a human blood plasma-derived protein corona on said uptake was assessed. EVs were isolated from collected PC-3 cell culture medium using differential centrifugation. Experiments were performed separately for MVs (20000xg EV-fraction) and EXOs (110000xg EVfraction). SDS-PAGE analysis revealed adsorption of plasma proteins to EVs, following their exposure to plasma. Prior to uptake experiments DiO-labelled EVs were either incubated or not incubated in plasma. Plasma incubation lasted overnight. PC-3 cells were then treated with either of the two EV-preparations. Following incubation, EV uptake was assessed using confocal microscopy by determining the percentage of positive fluorescent cells in cell cultures. Pre-study plasma incubation resulted in a reduced or unchanged uptake of MVs and in a reduced uptake of EXOs, when compared to their native counterparts. In conclusion, the plasma-derived protein corona was shown not to improve EV uptake. It is worth noting that the current study limits itself to the use of PC-3-derived EVs and PC-3 cells as recipient cells in uptake experiments.
  • Viskari, Ansa (Helsingfors universitet, 2012)
    The purpose of this study was to investigate how the mixing time of the magnesium stearate affects on the compressibility of partially pregelatinized maize starch. Pregelatinized maize starch is used in pharmaceuticals as a filler, binder and as disintegrant. Because pregelatinized maize starch has lubricant characteristics itself, it is known to be sensitive for the amount and the mixing time of magnesium stearate. The aim is that magnesium stearate is not totally homogenously mixed on the powder surfaces so that even, clean powder surfaces exist. Homogeneous mixing means that particles are coated with magnesium stearate, which as a hydrophobic ingredient prevents bond formation between plastically and elastically behaving particles. Too much magnesium stearate and/or too long mixing time may cause weakening of tablet tensile strength, laminating and capping. The weakening of the tensile strength of the tablet increases friability, which causes problems during packaging process and the transportation. Too much magnesium stearate may also lengthen the disintegration time and slow down the dissolution. The aim of this study was to compare four different brands of pregelatinized maize starch. The purpose was to find differences affecting the compressibility behavior. Also the effect of the mixing time of magnesium stearate for compression behavior of masses were studied. The brands investigated were C*PharmGel DC 93000, Lycatab® C, Starch 1500® and SuperStarch 200®. First mentioned was a reference product which is not manufactured any more. There was only one batch of the reference product but three batches from other products to be able to investigate also batch to batch variation. The characteristics studied from pregelatinized starch samples were bulk density, apparent density and true density, flowability, moisture sorption, moisture content, pH value, swelling volume and particle size. Also NIR, FTIR and Raman spectroscopy and X-ray powder diffraction method were used. Weight, tensile strength, dimensions, friability, disintegration time and moisture sorption were studied for tablets. The compressibility of the mass and elastic behavior of tablets was studied. Pictures of the tablets were also taken by scanning electron microscope. When the mixing time of magnesium stearate was increased from 2 minutes to 5 minutes, the compression pressure needed for pressing tablets for 80 N strength increased 200-700 N depending on the brand of pregelatinized maize starch. Based on the results the best alternative to replace C*PharmGel DC 93000 was chosen to be SuperStarch 200®. Scanning electron microscope pictures showed that C*PharmGel DC 93000 deviates from other qualities studied by being roundish and regular in shape. SuperStarch 200® and Starch 1500® reminded remarkably each other. Lycatab® C was the biggest in particle size and very irregular in shape. The differences found in tabletting followed the expectations based on the SEM-pictures. SuperStarch 200® showed to best compressibility in lowest strain strength and after C*PharmGel DC 93000 it was least sensitive for mixing time of the magnesium stearate. It also has least elastic recovery. The differences between SuperStarch 200® and Starch 1500® in compression properties were moderate but clear. Lycatab® C had clearly the weakest compression properties.
  • Antila, Hanna (Helsingfors universitet, 2012)
    Tissue plasminogen activator (tPA) is a serine protease that cleaves the inactive plasminogen to a broad-spectrum protease plasmin. Plasmin is involved in the degradation of blood clots by breaking down the fibrin network. In addition to it's role in the fibrinolytic system, tPA participates in the functions of the central nervous system. tPA is expressed in several brain areas and has been shown to be involved in neuronal plasticity. tPA's effects on brain plasticity are mediated in part via degradation of extracellular matrix proteins, but mainly via processing of brain-derived neurotrophic factor (BDNF). Plasmin cleaves pro-BDNF into BDNF that serves as primary endogenous ligand for TrkB neurotrophin receptor. TrkB signalling is strongly associated with the regulation of neuronal plasticity such as neurogenesis, synaptogenesis and long-term potentiation (LTP). On the contrary, pro-BDNF binds and activates p75 neurotrophin receptor that regulates many distinct, even opposite, effects on neuronal plasticity such as long-term depression and synapse refraction. Enhancement of brain plasticity is considered to be important for the therapeutic effects of antidepressant drugs and this is at least partially mediated via BDNF. Antidepressants activate TrkB receptors and increase BDNF protein levels in the rodent brain but the mechanism behind this remains obscure. Given that tPA is an important factor in the processing of BDNF, it is a possible mediator for antidepressants' neurotrophic effects. The effects of antidepressants on tPA activity have been previously studied only in the blood circulatory system. The aim of the experimental part of this Master's thesis was to examine the effects of antidepressant fluoxetine on tPA activity and protein levels in mouse hippocampus. Also the effects of fluoxetine on BDNF-TrkB signalling were studied. Fluoxetine was administered to mice acutely (30 mg/kg, i.p., 1 h) and chronically (0,08 mg/ml in drinking water, 3 weeks). tPA activity was studied using SDS-PAGE - and in situzymographies. TrkB activation, tPA and BDNF protein levels were measured using western blot. BDNF protein levels were also examined with ELISA method. No changes in tPA activity were found after acute fluoxetine treatment. In line with this result is the observation that also the BDNF levels remained unchanged. However, TrkB receptor activity was increased in fluoxetine treated mice. It seems possible that BDNF is not involved in the TrkB activation caused by acute fluoxetine treatment. Chronic fluoxetine treatment caused a significant increase in the BDNF protein levels compared to water-drinking control mice. This was not, however, associated with significant changes in TrkB activity. No changes in tPA activity were observed, which suggests that tPA is not involved in the increase of BDNF levels after chronic fluoxetine treatment. Interestingly, tPA antibody detected three distinct proteins in western blot of whose levels acute fluoxetine treatment regulated. However, more studies are needed to identify these proteins and to reveal the significance of such an effect of fluoxetine. According to this study, neither acute nor chronic fluoxetine treatment affects tPA activity in mouse hippocampus. However, environmental enrichment has been shown to enhance tPA activity and produce similar neurotrophic effects as chronic fluoxetine treatment. Therefore the result of this study concerning effect of chronic antidepressant treatment on tPA activity should be verified.
  • Yli-Mannila, Hanna (Helsingfors universitet, 2009)
    Stakes ja Lääkehoidon kehittämiskeskus määrittelevät lääkitysturvallisuuden lääkkeiden käyttöön liittyväksi turvallisuudeksi, joka voi vaarantua missä tahansa lääkehoidon vaiheessa. Yksi lääkitysturvallisuuteen vaikuttava lääkehoidon osa-alue on lääkeneuvonta. Koska lääkkeitä käytetään ammattilaisilta saatujen ohjeiden perusteella, on lääkeneuvonnan kehittäminen tärkeää. Lääketietouden lisäämisen ohella lääkeneuvonnalla voidaan vaikuttaa lääkkeiden käyttäjien asenteisiin. Lapsen lääkitykseen kohdistuva lääkeneuvonta on haasteellista, sillä huomio täytyy kiinnittää sekä lapseen että vanhempiin. Tutkimuksessa selvitettiin sairaalasta kotiutettavan lapsipotilaan lääkeneuvonnan sisältöä ja kehitystarpeita sekä lääkeneuvonnan toteuttamiseen vaikuttavia asioita. Lisäksi tutkimuksen avulla selvitettiin vanhemmille kotiutuksen jälkeen ongelmallisia lääkehoitoon liittyviä asioita. Tutkimusaineistona käytettiin HUS:n Lastenklinikalla työskentelevien sairaanhoitajien teemahaastatteluja. Toisena aineistona käytettiin vanhempien osastolle soittamia, lasten kotiutuksen jälkeistä lääkehoitoa koskevia puheluita. Aineiston keräys tapahtui hoitajien avulla heidän kirjatessaan tarkoitusta varten muodostettuun lomakkeeseen ylös ne lääkehoitoa koskevat kysymykset, joita vanhemmat soittavat osastoille kotiutuksen jälkeen. Keräykseen osallistuivat HUS:n infektio-osaston, lastentautien osaston ja munuais- ja elinsiirto-osaston hoitajat. Syyskuun 2009 aikana suoritettuihin teemahaastatteluihin osallistui 10 infektio-osaston sairaanhoitajaa. Kuten aiemmissakin tutkimuksissa on todettu, sairaanhoitajilla on suuri vastuu sairaalassa annettavasta lääkeneuvonnasta. Tutkimustulosten mukaan sairaanhoitajat pyrkivät kattavaan lääkeneuvontaan. He painottavat erityisesti perheen pärjäämistä kotona havainnollistamalla antamaansa neuvontaa kirjallisen materiaalin ja antotekniikkaan liittyvän opetuksen avulla, sekä kiinnittämällä huomiota perheen asenteisiin lääkehoitoa kohtaan. Esteinä lääkeneuvonnan toteuttamiselle koettiin sekä vanhempien että hoitajien kiire, joidenkin vanhempien vääränlainen asenne, hoitajien ja vanhempien kielimuurista johtuvat kommunikaatio-ongelmat, neuvontatilan rauhattomuus sekä hoitajille suunnattujen yhteisten ohjeistuksien puute. Lääkeneuvontaan liittyviksi kehityskohteiksi nousivat sekä vanhemmille että hoitajille suunnatut kirjalliset ohjeet, lääkkeisiin liittyvä lisäkoulutus, neuvonnan ajoitus, neuvontatila ja osastofarmaseutin osallistuminen neuvontaan. Kahden kuukauden tutkimusjakson aikana loka-marraskuussa 2009 kirjattiin osastoilla yhteensä 26 kotiutetun potilaan lääkehoitoa koskenutta puhelua. Infektio-osastolle tulleet yhteydenotot (n=7) koskivat pääasiassa lääkkeen annosteluaikoja ja kuurin tarkkaa kestoa. Yksi puhelu koski haittavaikutuksen ilmaantumista ja yksi antotekniikkaa. Lastentautien osastolle tulleista puheluista (n=11) kymmenen koski insuliinien annostusten muuttamista verensokeriarvojen korjaamiseksi. Munuais- ja elinsiirto-osastolle vanhemmat soittivat yleensä kysyäkseen annosteluun liittyviä erityisasioita, kuten annostelua oksennuksen, väärän lääkkeenantoajan ja yliannoksen jälkeen.Munuais- ja elinsiirto-osastolla puheluita kirjattiin yhteensä kahdeksan. Tutkimuksen mukaan hoitajien antama lääkeneuvonta on perusteellista. Lääkehoitoon liittyviä asioita jää kuitenkin vanhemmille epäselväksi, vaikka huomiota kiinnitetään niihin asioihin, joissa vanhemmat tarvitsevat eniten tukea. Osastoille kehitetyn lääkeneuvontamallin tarkoitus on auttaa hoitajia lääkeneuvonnan toteutuksessa, mutta huomionarvoista olisi myös osastofarmaseutin mahdollisuus osallistua neuvontatilanteeseen. Jatkotutkimuksissa olisi hyvä selvittää, millä tavalla neuvontamallia hyödynnetään neuvonnan toteuttamisessa. Erityisesti lapsille suunnattua lääkeneuvontaa on kehitettävä.
  • Tyyskä, Miia (Helsingfors universitet, 2009)
    Diabeetikoiden määrä lisääntyy jatkuvasti. Samalla hoitokulut ovat kasvaneet merkittävästi. Paras tapa hillitä kustannusten kasvua on hoitaa diabetesta mahdollisimman hyvin. Näin voidaan ehkäistä myös diabetekseen liittyvien oheissairauksien syntyä. Diabeteksen hoidossa on tärkeää kiinnittää huomiota hoidon jatkuvuuteen ja potilaan hoitoon sitoutumiseen. Apteekin henkilökunnan asema on noussut yhä keskeisemmäksi diabeetikon hoitoon sitoutumisen edistäjänä. Tämän pro gradu -tutkielman tavoitteena oli selvittää, mikä on apteekin farmaseuttisen henkilökunnan rooli diabetespotilaan hoitoon sitouttamisessa, elämäntapamuutosten toteuttamisessa ja niiden pysyvyyden varmistamisessa. Asiaa tarkasteltiin voimaantumisen teorian näkökulmasta. Tarkoituksena oli selvittää, miten voimaantuminen yksilössä tapahtuu, miten sitä voidaan apteekkineuvonnalla edistää ja mikä on apteekin rooli ulkopuolisena voimaannuttajana. Tämän tutkimuksen aineisto on kerätty Mäntyharjun Havu apteekissa ja se on osa laajempaa tutkimusta, jonka päätavoitteena on kehittää ja testata apteekkeihin soveltuva yksilökeskeinen toimintamalli tyypin 2 diabeteksen hoidon tukemiseen. Toimintamalli perustuu säännöllisiin neuvontatapaamisiin apteekissa. Pro gradu -tutkielmaan analysoitavaksi valittiin tutkimusjoukosta (n=19) ne, joilla tapahtui apteekkiohjelman aikana eniten positiivisia muutoksia yksilötasolla sekä elämäntapamittareilla mitatuissa arvoissa että kliinisissä parametreissa (n=4). Kvaliatiivisessa analyysissä käytettiin sekä deduktiivista että induktiivista lähestymistapaa. Vaikka diabeetikoilla oli tietoa sairaudesta ja elämäntapojen merkityksestä, niin käytännön tasolla jokainen henkilö kaipasi hoitoon ja erityisesti muutosten toteuttamiseen tukea ulkopuoliselta taholta. Apteekin rooli ulkopuolisena voimaannuttajana koettiin erityisen keskeiseksi. Tapaamiset loivat oikeanlaisen ympäristön ja ilmapiirin elämäntapamuutosten toteuttamiseen ja voimaantumisprosessin etenemiseen. Voimaantuminen ruokavaliomuutoksiin oli koko intervention aikana melko nousujohteinen prosessi. Sen sijaan voimaantuminen liikunnalliseen elämäntapaan oli aaltoilevaa. Apteekkitapaamiset sosiaalisena tapahtumana paransivat asiakkaan hoitoon sitoutumista. Asiakas koki, että häntä kohdellaan yksilönä kokonaisvaltaisesti. Voimaantuakseen yksilö tarvitsi aikaa. Vuoden mittaisen intervention aikana voitiin saavuttaa pysyviä muutoksia elämäntapoihin, mikäli yksilöllä itsellään oli halu ja motivaatio sitoutua tukiohjelmaan. Tämä tutkimus osoitti, että tämänkaltaista apteekkiohjelmaa tarvitaan. Nykyisessä kiireyhteiskunnassa ihmiset arvostavat, jos jollakin on aikaa paneutua yksilöön itseensä ja hänen sairautensa hoitoon kokonaisvaltaisesti.

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