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Jätevedenpuhdistamon käsittelymenetelmät eivät poista kokonaan lääkeaineita jätevedestä. Lääkeainejäämiä jää käsiteltyyn jäteveteen ja siksi niitä löytyy pintavedestä. Vielä ei ole täysin selvää, miten altistuminen lääkeainejäämille vaikuttaa ihmisten terveyteen ja ympäristön hyvinvointiin. Tämän pro gradu -tutkielman tavoitteena oli ensin tunnistaa vesiympäristölle haitallisimmat lääkeaineet ja sen jälkeen selvittää, mitkä olivat haitallisimpien lääkeaineiden käyttömäärät HUSin hoitoyksiköissä vuonna 2021. Tutkimuksen tuloksia on tarkoitus hyödyntää jatkossa HUSissa, kun arvioidaan mahdollisia toimenpiteitä ympäristöriskien vähentämiseksi. Tässä tutkimuksessa ympäristöriskiä mahdollisesti aiheuttavien aineiden arviointi koski lääkeaineita, joita käytettiin ihmisten hoitoon vuonna 2021 Suomessa. Tutkimukseen otettiin mukaan ne lääkeaineet, joista oli saatavilla riittävästi dataa riskiosamäärän laskemiseksi (kun ei otettu huomioon lääkeaineen poistumaa jätevedenpuhdistamolla). Tutkimuksen ulkopuolelle rajattiin lääkeaineiden yhdistelmävalmisteet, eläinlääkkeet, vitamiinit, elektrolyytit, aminohapot, peptidit, proteiinit, hiilihydraatit, lipidit, kasvirohdosvalmisteet ja rokotteet. Riskiosamäärä laskettiin 521 lääkeaineelle. Lääkeaineiden, joiden riskiosamäärä oli suurempi kuin 1, katsottiin aiheuttavan riskiä vesiympäristössä: mitä suurempi oli aineelle laskettu riskiosamäärä, sitä suurempi oli ympäristöriski. 521 lääkeaineesta 39 lääkeaineen riskiosamäärä ylitti raja-arvon 1. Lääkeaineet, joiden riskiosamäärä oli yli 10, olivat ibuprofeeni, dabigatraanieteksilaatti, sulfasalatsiini, estradioli, lerkanidipiini, sertraliini, abirateroni, amoksisilliini, rifaksimiini ja vankomysiini. Näiden kymmenen vesiympäristölle haitallisimmin lääkeaineen kulutus vuonna 2021 HUSissa laskettiin hoitoyksiköittäin, jotta saatiin selville, missä hoitoyksiköissä käytettiin näitä lääkeaineita eniten. Kulutuksen laskennassa ei otettu huomioon hoitoyksiköiden jo olemassa olleita lääkevarastoja, jotka oli hankittu ennen vuotta 2021, eikä lääkehävikkiä (lääkejätettä). Tämän tutkimuksen perusteella puuttuva data oli merkittävin tulosten luotettavuuteen vaikuttava tekijä, koska ne lääkeaineet, joista ei ollut riittävästi dataa käytettävissä, jouduttiin rajaamaan tutkimuksen ulkopuolelle. Siten vesiympäristölle haitallisimpien lääkeaineiden luettelosta saattaa puuttua sinne kuuluvia lääkeaineita. Dataa olisi tarvittu etenkin lääkeaineiden poistumasta jätevedenpuhdistamolla sekä tutkimuksesta poisjääneiden lääkeaineiden kulutuksesta, teoreettisista vuorokausiannoksista ja ennustetuista vaikutuksettomista pitoisuuksista. Koska kaikkia haitallisimpia lääkeaineita ei ole vielä selvitetty, lisää tutkimusta ja työkaluja tarvitaan ongelman ratkaisemiseksi

Cancer is a significant challenge for human health, causing approximately 9,6 million deaths annually. Critical problem in cancer treatments is chemoresistance, where cancer cells develop a resistance to the drugs used in the treatment. Chemoresistance has been studied for example with fluorescence spectroscopy and mass spectrometry. However, there are better options, as for example autofluorescence and photobleaching are a challenge in fluorescence spectroscopy and sample destruction in mass spectrometry. Vibrational spectroscopy is a label-free global method, which offers data extensively. The aim of this study is to evaluate the applicability of vibrational spectroscopy for studying chemoresistance using cisplatin sensitive TYK-nu cells and cisplatin resistant TYK-nu-CPR cells. Additionally, it is studied if there are difference on the spectra of a sample measured in liquid and dried. Infrared and Raman spectroscopy, which are used, are non-invasive and label free techniques that provide information about the biochemical structure of the samples. Techniques used in this study are infrared spectroscopy, Fourier transform Raman spectroscopy and confocal Raman spectroscopy. All techniques successfully distinguished the cell lines from each other according to the principal component analysis. Ratiometric analysis suggests a higher lipid to protein ratio in TYK-nu-CPR cells, which might be the separating factor between TYK-nu and TYK-nu-CPR cells. Liquid seemed to cause overlapping with peaks originating from the cell, and a large water related peak, which has to be taken into account if measuring cells in liquid.

Many pharmacies sell medicines through digital online pharmacy services but studies have shown that only a small fraction of the population are aware of and are using these. There is a need to investigate online pharmacy services and the obstacles they are facing. What should be considered are e.g. the legislation online pharmacies operate under, how the health competency of the population is taken into account in the design of the services, the skills and attitudes of the professionals toward digitalization and distance communication as well as practical circumstances. The aim of this study is to examine the online pharmacy operators' perceptions of the services they provide and how they are organized in Finland and Sweden. How the national regulations that exist regarding online pharmacies, and especially regarding medicine counseling at online pharmacies, are implemented in practice are also examined. The study was targeted to all pharmacy actors in Finland (72) and Sweden (11) who offer online pharmacy services and have reported this to the authorities. Of these, 20 (28 %) responded in Finland and 4 (36 %) in Sweden. The study was designed as a survey consisting of closed/structured- and open-ended questions. The survey questions were developed from previous literature on the subject and a background analysis of the online pharmacies' web pages to ensure the baseline and the relevance of the questions. The survey was available to respondents in digital format and the link was distributed through e-mail in Winter 2020. Data from the closed-ended questions were compiled and analyzed using Microsoft Office Excel, while data from the open-ended questions were analyzed through qualitative content analysis. In Finland, only a small fraction of the total turnover of pharmacies comes from the online pharmacy business. Many believe in an increased importance of online pharmacies when younger age groups begin to use pharmacy services more frequently and technology enables more versatile solutions. In both countries, the main age group using online pharmacies were 36-45 years. The goods that were purchased were mainly cosmetics, skincare and non-prescription drugs. In Sweden, customers suffering of long-term illness were also reported to be a large customer group. As distance communication methods, telephone and e-mail were mostly offered, but in Sweden also chat. About half of the staff had received special communication training for distance communication. Many respondents stated being unaware of the fraction of customers asking for medication counseling and whether this counseling is appreciated. In most cases, the operators stated that the legislation on online pharmacy services was adequate. There are differences in the possibilities of operating online pharmacies in different markets. In Finland, the online pharmacy business's share of pharmacies' total sales is very limited. In Finland there is hope that online pharmacies will become more important to younger customer groups in the future and that the technology is developed to enable more versatile services. Operators in Finland believe that the services should be more beneficial to customers in the future. In both countries an increase in the sale of prescription medication is expected. Online pharmacies have great opportunities to offer customers versatile means of communication for medication counseling, but there is still a need to investigate employees' skills in distance communication. The counseling situation at online pharmacies should be evaluated to ensure equivalency with the one at physical pharmacies.

Gut inflammation and permeability is speculated to play a major role in the pathophysiology of several human diseases. Signs of a low-grade gut inflammation in patients with type 1 diabetes (T1D) have been found. Focus of this study was to understand the role of gut inflammation and increased gut permeability in the development of diabetic complications, especially nephropathy. Approximately, one-third of Finnish patients with T1D develop kidney disease during their lifetime. Inflammatory mechanisms may have an essential role in the pathophysiology of the disease. Lipopolysaccharide, LPS, is found in the outer membrane of gram-negative bacteria. LPS activates innate immune system and triggers the activation of inflammatory cytokines, neutrophils and macrophages as well as many pathophysiological processes in vivo, for instance fever and endotoxic shock. Aim of this study was to establish a zebrafish gut inflammation model using fluorophore conjugated endotoxin, LPS. We hypothesized that delivery of LPS in addition to EDTA in the gut of zebrafish triggers inflammation and increased gut permeability which may lead to leakage of LPS to blood stream and potentially kidney injury. This novel zebrafish inflammation model could possibly be used for studying the pathophysiological mechanisms of gut inflammation and possible kidney injury as well as for screening new anti-inflammatory drugs. In addition, this animal model can be used for studying intestinal alkaline phosphatase (IAP) in reducing gut permeability and LPS-mediated kidney damage. IAP is an enzyme produced in small-intestinal epithelium. IAP can detoxify several bacterial endotoxins including LPS and thus protect against the induction of intestinal inflammation. LPS and EDTA were delivered in the gut of 6 days old zebrafish larvae using microgavage injection. Fluorescence microscopy imaging of live zebrafish enabled following the same individual at different timepoints after injections. Paraffin sectioning of the small larvae was promising for investigating the morphology and permeability of the gut as well as possible immunostaining for detection of IAP. L-phenylalanine was used for inhibition of IAP enzyme. Using the novel method of microinjection to gut on zebrafish larvae the timing and amount of delivered materials to gut can be controlled well. The anatomy and function of the gut in zebrafish is very similar to small intestine of mammals and the highly developed vertebrate immune system makes zebrafish an interesting model organism for studying gut inflammation and permeability. In addition, inflammatory processes can be visualized in live, intact transparent zebrafish larvae. However, the technique has a lot of challenges including small size of the fish and possible tissue damage of the fish while performing injection. More experiments need to be carried out to establish the model for drug screening. Also, along with microscopy images, a more precise way for quantification the gut permeability is needed. Based on the images it’s not yet possible to conclude whether LPS increased gut permeability or if IAP inhibition with L-phenylalanine worked in zebrafish larvae. Using adult zebrafish in the future will give more information about the chronic gut inflammation and development of possible kidney injury.

The α5 subunit of nicotinic acetylcholine receptors forms functional receptors with other subunits as a structural subunit. It affects the structural and functional properties of the nicotinic receptor by increasing calcium permeability and accelerating desensitization. In the mammalian brain, the α5 mRNA is widely expressed, mostly in substantia nigra pars compacta, ventral tegmental area and interpeduncular nucleus. Its protein has been identified in various distinct brain areas, such as striatum, cortex and medial habenula. In the dorsal striatum partaking in motor functions, the α5 subunit modulates the release of dopamine, thus it is believed to have an impact on motor function. In the experimental part of the thesis mice lacking the α5 subunit were injected unilaterally with neurotoxin 6-hydroxydopamine (6-OHDA) in the striatum. The purpose was to determine the importance of the subunit with regard to the lesion extent and motor function. The motor functions of α5-deficient and wild type control mice were assessed in amphetamine- and apomorphine-induced rotametry. After the tests the mice were euthanized and their substantia nigra and striatal brain samples were collected for further analysis. The number of dopamine cells in the medial and dorsal tier of substantia nigra were determined, so as to quantify the extent of the lesions and to explain the research group's previous finding about the α5-deficient mice spinning less ipsilaterally in amphetamine induced rotametry. The α5-deficient mice were found to turn less ipsilaterally compared to the control mice in the amphetamine-induced rotametry and in the apomorphine-induced rotametry, first less contralaterally and subsequently more contralaterally than the control mice. The results of male mice, that were less in number, were excluded from the results as the difference between genders was significant in the wild type mice in the amphetamine-induced rotametry. There was no significant difference in the number of remaining dopamine cells between the genotypes after the lesioning in either of the areas of interest. However, the wild type mice tended to have less cells remaining in the medial tier of the substantia nigra. The observed differences between the genotypes in the rotametries could be accounted by differences in the amount of dopamine released from striatal neurons or differences in striatal dopamine receptor quantities or function. The results support the hypothesis about the contribution of the α5 subunit containing acetylcholine nicotinic receptors in motor function.

lcohol addiction is a significant public health problem worldwide, and its treatment is extremely challenging. One major problem in the treatment of alcohol addiction are the later relapses to uncontrollable drinking. Approximately 60-70 % of addicts relapse to drinking within a year from the beginning of the treatment. The current treatment of alcohol addiction is based on a combined psychotheraphy and pharmacological treatments, but even at the best the efficacy remains quite modest. This is why further studies on the underlying mechanisms behind alcohol addiction and development of more effective pharmaceuticals to treat it are an important field of research. Chronic exposure to the rewarding effects of alcohol causes neurochemical adaptations in the brain reward system. These adaptations strive to restrain the recurring rewarding signals caused by alcohol and lead eventually to increased reward thresholds in the reward system. As the reward thresholds increase, the individual develops tolerance to the rewarding effects of alcohol, but also craving for the substance and a dysphoric mental state which are highlighted especially during periods of abstinence. It is known that the increase in reward thresholds is an important factor leading to relapses, but the exact nature of the neurochemical adaptations behind it are not known. According to recent studies dynorphin -peptides (DYN) and κ-opioid receptors (KOR) of the endogenous opioid system seem to have an important role in these neurochemical adaptations. It has been shown that chronic alcohol exposure increases the activity of DYN/ KOR -system especially within the nucleus accumbens (NAc), which is an essential structure of the brain reward system. The increased activity of the DYN/ KOR -system in the NAc has been shown to inhibit the development of rewarding signals. Previous studies have shown that inhibiting the increased activity of the DYN/ KOR -system with a selective KOR-antagonist, reduces voluntary alcohol intake and relapse-like alcohol seeking behavior during periods of abstinence, especially in physically addicted animals. In this study we studied the relapse-like alcohol drinking of Long-Evans rats in the alcohol deprivation effect (ADE) model. The effects of selective and long-acting KOR-antagonists, JDTic and nor-BNI, were tested on the ADE-effect which occurs after a period of deprivation. The ADE is defined as a transient increase in alcohol intake after a forced period of abstinence and it has been shown both in rodents with a history of alcohol consumption, and human alcohol addicts. In this study the rats were allowed to consume alcohol (10% ethanol-water solution) voluntarily during 90 minutes for 10 consecutive days after which followed a six days long deprivation period. According to results, both intra-accumbally (15 µg/ 0,3 µl/min/ site) or subcutaneously (10 mg/kg) administered JDTic decreased the ADEeffect significantly compared to vehicle, when administered 24 hours prior the end of the deprivation period. Also intra-accumbally administered nor-BNI (3 µg/0,3 µl/min/site) decreased the ADE-effect significantly compared to vehicle when administered 24 hour prior the end of deprivation. The results are in line with the theory that alcohol induces sensitization of the DYN/ KOR -system within the brain structures involved in reward. In theory it can be speculated that by suppressing the activity of the DYN/ KOR -system, KOR-antagonists can relieve craving for alcohol. This can be seen as a decrease in relapse-like consumption of alcohol. In conclusion, it can be suggested that by suppressing the increased activity of the DYN/ KOR -system induced by chronic alcohol exposure with a selective KOR-antagonist, like JDTic or nor-BNI, it could be possible to reduce the risk of relapse during abstinence and thus improve the efficacy of treatments for alcohol addiction.