Dissertations / Theses on the topic 'Farmakoloji'

Poslednjih godina je prisutan trend povratka prirodi i upotrebi biljnih lekova, kako u prevenciji tako i u lecenju razlicitih bolesti. Timijan (Thymus vulgaris L.) se u narodnoj medicini koristio u lecenju respiratornih oboljenja kao što su kašalj, bronhitis i astma. Rezultati novijih istraživanja pokazuju da timijan poseduje i druga potencijalno korisna farmakološka svojstva (antimikrobna, antiinflamatorna, antioksidativna, spazmoliticka, antidijabetesna i anksioliticka). Ciljevi ovog istraživanja su bili da se ispitaju farmakodinamske osobine preparata timijana, njihove interakcije sa lekovima koji deluju na centralni nervni sistem, uticaj na funkciju jetre i parametrem oksidativnog stresa kod životinja izloženih ugljentetrahloridu, kao sadržaj karvakrola i timola u sirupu timijna, pri razlicitim uslovima cuvanja. U farmakodinamskim ispitivanjima kao eksperimentalne životinje korišceni su miševi soja NMRI, a u svim drugim ispitivanjima pacovi soja Wistar. Tinktura timijana je primenjena u dozi od 0,4mk/kg, a sirup u dozi od 12,08 ml/kg, na miševima. Primenjene doze na pacovima su bile 0,18 ml/kg za tinkturu i 5,6 ml/kg za sirup timijana. Za ispitivanje analgetickog dejstva korišceni su metod vrele ploce i test sircetne kiseline. Za procenu motorne koordinacije korišcen je test rotirajuceg štapa, a za procenu hipnotickog delovanja mereno je vreme spavanja. Prilikom ispitivanja uticaja preparata timijana na farmakokinetiku paracetamola, odre_ivana je koncentracija ovog leka HPLC metodom, a nakon toga su odreeni farmakokinetski parametri paracetamola. Antioksidantna aktivnost preparata timijana odre_ivana je pomocu in vitro i in vivo testova. Nakon žrtvovanja životinja ra_ena je histopatološka analiza jetrenog tkiva, a u serumu su odre_ivani biohemijski parametri, kao i pokazatelji bubrežene i jetrene funkcije. Sadržaj timola i karvakrola i sirupu timijana odre_en je GC/MS metodom. Sirup i tinktura timijana su pokazali analgeticki efekat u testu vrele ploce, kao i smanjenje broja grceva izazvano primenom sircetne kiseline. Sedmodnevna primena preparata timijana smanjila je analgeticko dejstvo kodeina, a pojacala analgeticki efekat paracetamola. Sirup timijana je potencirao diazepamom izazvan poremecaj motorne koordinacije. Ispitivanjem uticaja preparata timijana na hipnoticko delovanje pentobarbitala, postignuti su razliciti rezultati u zavisnosti od dužine trajanja pretremana. Sedmodnevna primena timijana je produžila vreme trajanja spavanja, dok je jednokratna primena timijana skratila vreme trajanja spavanja. Nakon i intravenske i peroralne primene paracetamola, grupe životinja koje su bile pretretirane preparatima timijana imale su krace poluvreme eliminacije i vecu konstantu eliminacije. Upotreba samo preparata timijana nije imala uticaj na biohemijske i histološke promene jetrene funkcije. S druge strane, upotreba tincture timijana u kombinaciji sa ugljen-tetrahloridom dovela je do porasta vrednosti AST i ALT enzima u serumu, dok je sirup timijana u kombinaciji sa ugljentetrahloridom smanjio aktivnost aminotransferaza. Najvece odstupanje u koncentracijama aktivnih komponenti timola i karavkrola, pokazali su sirupi cuvani na sobnoj temperaturi (20°C), u sekundarnoj ambalaži i na svetlom mestu. Rezultati dobijeni u toku ovog istraživanja ukazuju da preparati timijana uticu na farmakodinamske osobine kodeina, paracetamola, diazepama i pentobarbitala, kao i na farmakokinetiku paracetamola. Upotreba preparata timijana ispoljila je analgeticki efekat i umanjila posledice izloženosti oksidativnom stresu. Uslovi cuvanja sirupa timijana uticali su na njegovu stabilnost.
In recent years is present trend of return to nature and the use of herbal medicines in prevention and treatment of different diseases. Thyme (Thymus vulgaris L.) was used in folk medicine in the treatment of respiratory diseases such as cough, bronchitis and asthma. The new research results have demonstrated that thyme has many others potentially useful pharmacological properties (antimicrobial, antiinflammatory, antioxidant, antispasmodic, antidiabetic and anxiolytic). The aims of this research were to determine the pharmacodynamic properties of thyme preparations and their interactions with central nervous system drugs, influence on liver function and oxidative stress parameters of animals exposed to carbon tetrachloride, as well as concentration of thymol and carvacrol in thyme syrup, at different storage conditions. In pharmacodynamics examination as experimental animals were used NMRI mice, while in all other test were used Wistar rats. Applied dose of thyme tincture was 0.4 ml/kg and of syrup 12.08 ml/kg, for mice. For rats, applied doses of tincture and syrup were 0.18 ml/kg and 5.6 ml/kg, respectively. The analgesic activity was examined by the hot plate test and acetic acid test. The Rotarod test was used to evaluate the motor coordination and to evaluate hypnotic activity sleeping time was mesaured. In order to examine the influence of thyme preparations on pharmacokinetics of paracetamol, the concentracion of this drug was measured by HPLC metods, and after that pharmocokinetic parameters of paracetamol were determined.The antioxidant acivity of thyme preparations was determined by using in vitro and in vivo tests. After animals sacrificing, histopathological analysis of liver tissue were peroformed, in serum were determined biochemical parameters and renal and hepatic function parameters. Quantification of thymol and carvacrol in syrup was carried out by GC/MS method. Thyme syrup and thyme tincture exhibited analgesic activity in hot plate test and reduced the number of writhes induced by acetic acid. Seven-day pretreatment with thyme preparations reduced analgesic activity of codeine and increased analgesic effect of paracetamol. Thyme syrup potentiated diazepam induced motor coordination impairment. Examining the impact of thyme preparations on hypnotic effect induced by pentobarbital, different results were achieved depending on the duration of pretreatment. Seven-day pretreatment with thyme had prolonged the sleeping time, while after single dose of thyme the sleeping time was decreased. After intravenous and after oral administration of paracetamol, groups pretreated with thyme preparations had decreased elimination half-life and increased elimination constant rate. Administration of thyme preparations alone did not change biochemical nor histological markers of hepatic function. On the other hand, co-administration of thyme tincture and carbon tetrachloride resulted in exacerbation of AST and ALT values in serum, while thyme syrup in coadministration with carbon tetrachloride managed to reduce activities of aminotransferases. The concentration of major active compounds, thymol and carvacrol, was mostly changed when syrups were stored at room temperature (20°C), in secondary containers and in light place. Results obtained in this study demonstrated that thyme preparations do affect pharmacodynamic properties of codeine, paracetamol, diazepam and pentobarbital and pharmacokinetics of paracetamol. Administration of thyme preparations exhibited analgesic activity and reduced the effects of exposure to oxidative stress. Storage conditions of thyme syrup did affect its stability.

Upotrebom 50% etanola i 50% acetona kao ekstragensa izvršena je ekstrakcija delova ploda kestena: srž ploda, braon spoljna kora ploda, crvena unutrašnja kora ploda, ceo plod (bez ježevica), kao i drveta: lišće, resa, ježevice, stara i mlada kora stabla. Ispitivani su pitomi kesten, lovranski marun i kalemljeni italijanski marun. Nakon određivanja prinosa suvog ekstrakta, primenom standardnih spektrofotometrijskih metoda određen je sadržaj fenolnih jedinjenja, flavonoida i kondenzovanih tanina. Iako je primenom 50% acetona kao ekstragensa dobijen veći prinos fenolnih materija, flavonoida i kondenzovanih tanina, za proizvodnju ekstrakata se preporučuje 50% etanol kao ekstragens, jer obezbeđuje sasvim zadovoljavajuće rezultate, a prihvatljiviji je sa aspekta znatno niže toksičnosti.Ekstrakt lista lovranskog maruna i rese pitomog kestena roda 2007. godine štite eritrocite od hemolize izazvane H₂O₂.Glavne komponente hidrolizata nakon metanolizacije ekstrakata su dimetil estar dehidrodigalne kiseline i metil estar dilaktona elaginske i valoneinske kiseline. Kvantitativnom LC/MS i HPLC/DAD analizom najveći sadržaj elagitanina je utvrđen za ekstrakt ježevica (170,6 mg/g ekstrakta).Izvšeno je ispitivanje antioksidativne aktivnosti ekstrakata u odnosu na 1,1-difenil-2-pikrilhidrazil (DPPH), hidroksi (^•OH) i superoksidni anjon (^•O₂^-) radikale. Kapacitet ekstrakata za transformaciju organskih hidrofilnih radikala, ispitan kao sposobnost redukcije spin probe Tempon je najveći za ekstrakt rese pitomog kestena (A = 18,1%), dok ekstrakti lista pitomog kestena i mlade kore drveta ispoljavaju slabu aktivnost. Ispitivanje protektivnog delovanja ekstrakata u odnosu na UV zračenje određeno je kao sposobnost uklanjanja ^•OH i ^•O₂^- radikala nastalih nakon zračenja. Ekstrakti koji ispoljavaju pozitivne, ali relativno niske RI vrednosti za proizvodnju obe vrste radikala ^•OH i ^•O₂^- su spoljna braon kora lovranskog maruna, resa sa kalemljenog italijanskog maruna i list lovranskog maruna. Negativne RI vrednosti dobijene za ostale ekstrakte ukazuju na prooksidativnu aktivnost u vodenom rastvoru izloženom UV zračenju. Ekstrakti rese, lista i ježevica ispoljavaju aktivnost u cilju preveniranja/otklanjanja lipidne peroksidacije membrane eritrocita.Ispitivanjem in vitro antioksidativne aktivnosti primenom MTT testa je utvrđeno da ekstrakti rese i ježevica pitomog kestena i lista lovranskog maruna imaju izuzetno visoku antioksidativnu aktivnost u ćeliji. Naročito je povoljna činjenica da deluju u niskim koncentracijama (0,02 mg/ml). Antimikrobna aktivnost ekstrakata C. sativa određena je u odnosu na (G+) bakterije: S. aureus, S. lutea, B. cereus, L. lactis ssp. lactis, M. pyrogenes var. albus, kao i na (G-) bakterije: P. mirabilis i S. typhimurium. Značajnu antimikrobnu aktivnost daju ekstrakti kore drveta, ježevica i spoljne braon kore ploda. Ekstrakti srži ploda i celog ploda nisu ispoljili antimikrobnu aktivnost. Postoji značajna i jako značajna korelacija između antimikrobne aktivnosti ekstrakata, kao i antimikrobne i antioksidativne aktivnosti u odnosu na superoksid anjon radikale. Ekstrakti lista, ježevica, spoljne braon i unutrašnje crvene kore ploda, kao i kore drveta C. sativa Mill. dobijeni primenom 50% etanola kao ekstragensa predstavljaju značajan izvor komponenata sa farmakološkim delovanjem u cilju smanjenja nivoa oksidativnog stresa, poseduju visok kapacitet sprečavanja lipidne peroksidacije, deluju u pravcu preveniranja/otklanjanja lipidne peroksidacije i zaštite membrane eritrocita, imaju visoku in vitro antioksidativnu aktivnost, a ispoljavaju i značajnu antimikrobnu aktivnost.
Parts of chestnut such as: seeds (without spiny burs), peeled chestnut, red internal seed coat and brown seed coat, as well as parts of the trees: leaf, catkin, spiny burs, young and old chestnut bark have been extracted by 50% ethanol and 50% acetone as an extragent. Three cultivars of Castanea sativa Mill.: sweet chestnut, Lovran's marrone and grafted Italian marrone were examined. After determination of the yield of dry extract, the content of total phenolic compounds, flavonoids and condensed tannins are determined by application of standard spectrophotometrics methods. Although, the highest content of total phenolics, flavonoids and condensed tannins are obtained by 50% acetone as extragents, for production of extracts 50% ethanol is more suitable, regards much lower toxicity.Extracts of the leaf of Lovran's marrone and catkin of sweet chestnut native in 2007, protect erythrocytes from hemolysis provoked by H₂O₂.Dehydrodigallic acid dimethyl ester, ellagic acid and valoneic acid dilactone methyl ester are the main compounds in all hydrolysates after methanolisation. The highest content of ellagitannin was detected in extract of spiny burs (170.6 mg/g extract), by application of quantitative LC/MS and HPLC/DAD analysis.The examination of antioxidant activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (^•OH) and superoxide (^•O2^-) radicals have been done. Capacity of extracts for removal of organic, hydrophilic radicals, exanimate as potential of reduction of spin probe Tempon is highest in extract of catkin of sweet chestnut (A = 18.1%), while extracts of catkin, leaf and spiny burs almost have no antioxidative activity. The evaluation of UV-protective activity of extracts is determinated as capacity for removal of ^•OH and O₂^- radicals generated after irradiation. Extracts which showed positive, but relative low RI values for production of both radical species, OH and ^•O₂ radicals, are brown seed coat of Lovran's marrone, catkin of grafted Italian marrone and leaf of Lovran's marrone. Negative RI values obtained for other extracts show that these have prooxidative activity in aqueous solution exposed to UV radiation. Extracts of catkin, leaf and spiny burs expressed activity to prevent/remove lipid peroxidation in the membrane of erythrocytes.Examination of antioxidant activity in vitro by application of MTT test have been detected especially high antioxidant activity of extracts of catkin, spiny bur of sweet chestnut and leaf of Lovran’s marrone in the cell. Particularly is favorable that extracts acting in low concentration (0.02 mg/ml). Antimicrobial activity of extracts of C. sativa was determinated against Gram-positive bacteria: S. aureus, S. lutea, B. cereus, L. lactis ssp. lactis, M. pyrogenes var. albus, as well as Gram-negative bacteria: P. mirabilis and S. typhimurium. The significant antimicrobial activity shows extracts of bark, spiny burs and brown seed coat. Extracts of peeled chestnut and seeds didn’t show any antimicrobial activity. The very significant and significant correlation existed between antimicrobial activity of extracts, as well as antimicrobial activity and scavenging of ^•O2^- radical. Extracts of leaf, spiny burs, brown seed coat and red internal seed coat, as well as bark of C. sativa Mill. produced by 50% ethanol as extragent represent important resource of components with pharmacological activity in reducing level of oxidative stress, possess high activity to prevent/remove lipid peroxidation and protection of the membrane of erythrocytes, have high in vitro antioxidant activity, and also express significant antimicrobial activity.

Background: Statins are among the most used drugs in Sweden. There are currently four statins available on the Swedish market; atorvastatin, simvastatin, pravastatin and rosuvastatin. Statins act by inhibiting 3-hydroxy-3-methylglutaryl-coenzyme-A reductase, also known as HMG-CoA-reductase. HMG-CoA-reductase is the rate-limiting enzyme of the cholesterol synthesis. Decreased hepatic cholesterol leads to increased low-density lipoprotein (LDL) clearance from plasma to liver cells. Having a high level of LDL cholesterol is potentially dangerous as it can lead to atherosclerosis and cardiovascular disease. Statins significantly reduce cardiovascular morbidity and mortality in patients with and without coronary heart disease. Purpose: The aim of this work was to investigate if there are any differences between the different statins according to contemporary evidence. Method: This is a literary analysis. Studies included were searched from PubMed. A total of five studies were included. Results: The result of this study indicates rosuvastatin to be most efficacious in lowering LDL cholesterol (LDL-C), triglycerides and total cholesterol. It also improved the high-density lipoprotein cholesterol (HDL-C) better than atorvastatin, simvastatin and pravastatin. Study 1 showed rosuvastatin to lower LDL-C with statistical significance (P<0.001) across dose ranges (10-40 mg) after 12 weeks. Study 2 compared a dose ratio of 1:2 between rosuvastatin and simvastatin in lowering LDL-C with a 3.24% (95% CI 4.10 to 2.38) favor of rosuvastatin. Study 4 compared effects of atorvastatin 80 mg and rosuvastatin 20 mg in patients with ST elevation myocardial infarction in a 4-week therapy. Rosuvastatin 20 resulted in a 35% compared with atorvastatin 80 mg 34% (P=0.59) reduction in LDL-C levels. Study 1 demonstrated rosuvastatin to improve HDL-C levels in daily doses of 40 mg with statistical significance compared with atorvastatin, simvastatin and pravastatin. Study 1 demonstrated rosuvastatin to lower total cholesterol with statistical significance (P<0.002) across doses compared with atorvastatin, simvastatin and pravastatin. Study 1 also demonstrated rosuvastatin to lower triglycerides more. In daily doses of 40 mg rosuvastatin had a statistical significance (P<0.002) versus simvastatin and pravastatin, but not atorvastatin. The P value between rosuvastatin and atorvastatin was not mentioned, neither the P-value between atorvastatin, simvastatin and pravastatin. Conclusion: Statins are not equipotent. Rosuvastatin showed greater results in reducing LDL-C, triglycerides and total cholesterol with no increased risk of adverse events compared with atorvastatin, simvastatin and pravastatin. Rosuvastatin still lacks in clinical experience which makes needs for further studies on this topic.

Introduktion: ADHD är en uppmärksamhet -och hyperaktiv störning, där 90 % i Sverige behandlas med det farmakologiska läkemedlet Metylfenidat (MPH). En stor andel med ADHD utvecklar psykiatrisk komorbiditet i form av psykisk ohälsa (frånvaro av mentalt välbefinnande). Patofysiologin hur MPH påverkar hjärnan är okänd och behandlingsformen har diskuterats som bristfällig på grund av förekommande bieffekter. Fysisk aktivitet har föreslagits som ett hälsosammare behandlingsalternativ, då evidens visat att fysisk aktivitet kan förbättra ADHD-symptom och samsjuklighet av psykisk ohälsa. Det finns inga studier som undersökt hur individer med ADHD upplever nuvarande- och önskar behandling. Syftet med denna studie blev därför att undersöka upplevda erfarenheter av ADHD och hur det farmakologiska läkemedlet MPH och fysisk aktivitet upplevs ha påverkat diagnosens symptom och individernas psykiska hälsa samt att ta reda på vilken behandling individerna önskar utifrån deras livserfarenheter. Metoden utgick från ett fenomenologiskt ramverk med hjälp av en deskriptiv kvalitativ tvärsnittsstudie, där data extraherades genom en innehållsanalys med en induktiv ansats. Urvalet selekterades genom snöbollsurval, där inklusionskriteriet var att deltagarna blivit diagnostiserad med ADHD och har erfarenheter av MPH och fysisk aktivitet. Resultatet visade att symptom för ADHD var problematik med koncentration-, hyperaktivitet-, uppmärksamhet- och systematiska svårigheter, där symptom försämrat den psykiska hälsan. Det framgick att både MPH och fysisk aktivitet effektiviserade symptom mot koncentration, hyperaktivitet och uppmärksamhet, varav KBT (kognitiv beteendeterapi) förbättrade systematiska svårigheter. Vidare bidrog MPH med fysiska, mentala och beteendeföränderliga bieffekter som försämrade den psykiska hälsan. Fysisk aktivitet visade däremot förbättra den psykiska hälsan, där inaktivitet framgick som en central nackdel för förvärring av symptom och psykisk hälsa. Deltagarnas önskan var att behandling bör uppföljas och baseras på en överenskommelse som anpassas efter individuella svårigheter och hälsotillstånd. Slutsatsen med studien kom fram till att behandlas med fysisk aktivitet i kombination med KBT skulle vara hälsosammare som första behandling mot samtliga ADHD-symptom och psykisk hälsa. I andra hand var det deltagarnas önskan att applicera MPH vid akut behov, och är därav förslag att forska fram MPH som engångsbehandling som kan användas vid inaktiva tillstånd. Studien kom fram till att inkludering av fler idrottsvetenskapliga tjänster till vård och skola behövs för mer kompetens om vikten av fysisk aktivitet för individer som har ADHD.
Introduction: ADHD is an attention-deficit/hyperactivity disorder, where 90% in Sweden is treated with the pharmacological drug Methylphenidate (MPH). People diagnosed with ADHD develop often psychiatric comorbidity in form of mental illness. The pathophysiology behind how MPH influence the brain is unknown and the treatment have been discussed as deficient due to side effects. Physical activity has been suggested as a healthier treatment option, as evidence shows that physical activity can improve ADHD-symptoms and comorbidity of mental illness. There are no studies that have invastigate how individuals with ADHD experience current- and desired treatment. The aim of this study was therefore to investigate the experiences of having ADHD and how the pharmacological treatment MPH and physical activity has affected the symptoms and mental health, and also find out what treatment the individuals want based on their life experiences. The method was derived from a phenomenological framework using a descriptive qualitative cross-sectional study. Data was analyzed with an inductive approach using an interpretive analysis. Participants was recruited through a snowball selection, where the inclusion criteria stated that participants been diagnosed with ADHD and have experience of MPH and physical activity. The results showed that the frequent occurring symptoms of ADHD were difficulties with concentration-, hyperactivity-, attention- and systematic symptoms, where symptoms had shown a negative effect on the mental health. It appeared that both MPH and physical activity made symptom improvement against concentration-, hyperactivity- and attention difficulties, while CBT (Cognitive Behavioral Therapy) improved systematic difficulties. MPH contributed to side effects that affected physiological-, psychological- and behavioral aspects, which all were perceived to have a negative influence on mental health. In contrast, physical activity improved mental health and showed no evidence of side effects, except when physical activity was absent from daily life. Inactivity appeard to be a central disadvantage for worsening symptoms and mental health. The participants desired that their treatment should be tailored to individual difficulties and health condition in a collaborative way and with follow-up from the health care worker. The study concluded that physical activity in combination with CBT would be a healthier as first treatment option to reduce symptoms of ADHD and improve mental heatlh. Additionally, the participants desired treatment with MPH in form of single doses for urgent need, and it is therefore to suggest development of such treatment option. Furthermore, the study proposes more services for sport science within healthcare and schools for containing applicable knowledge in physical activity for dose with ADHD.

Many blood cell mechanisms in the human body are working all the time to maintain haemostasis in the blood vessels. Once a wound arises platelets are alerted via different substances to cover the wound and prevent loss of blood. Most of the times these mechanisms do stop the blood, and further heal the wound. During other circumstances the platelet-covering continues to form a thrombus, preventing the blood to flow and instead causes myocardial infarction or stroke. There are several risk factors triggering development of circulatory diseases such as obesity, lack of exercise, smoking, infection and stress.

This thesis describes the interaction between the two platelet thrombin receptors PAR1 and PAR4, together with the interaction of the oral pathogen Porphyromonas gingivalis (with thrombin-like gingipains), and the cross talk with the stress hormone epinephrine and its α_2A adrenergic receptor. Until now PAR1 is thought to be the most important thrombin receptor due to its high affinity for thrombin. From a phylogenetical and patophysiological point of view there must be a reason why platelets express two different thrombin receptors. Today PAR4 is considered less important, but this thesis implies that PAR4 plays an important role in platelet signaling and haemostasis.

The results show that bacteria pre-stimulated platelets, followed by epinephrine gives a strong and full aggregation and calcium mobilization, in both aspirinated and non-aspirinated human platelets. The amount of bacteria does not itself, or epinephrine alone give aggregation or calcium mobilization. This mechanism is dependent on both Rgp type gingipain released from P. gingivalis, and PARs in an interaction with the α_2A adrenergic receptor.

Further, results reveal that PAR4 interacts and cross talks with the platelet α_2A-adrenergic receptor in aspirinated platelets. Neither of the two platelet purinergic P2Y-receptors (P2Y₁₂ and P2Y₁) contribute to this action, but the purinergic P2X₁ does. In aggregation studies a low dose of PAR4 activating peptide (AP), but not PAR1-AP, followed by epinephrine results in a strong aggregation and in a calcium mobilization. ATP secretion measurements did reveal that ATP was released during epinephrine stimulation, which indicate that ATP and P2X₁ have a key role in this event. By blocking P2X₁ both aggregation and calcium mobilization were abolished, but not by blocking P2Y₁₂ and P2Y₁. Inhibition of PI3-kinase, both epinephrine-induced calcium mobilization and aggregation were significant reduced. In non-aspirinated platelets PAR1 synergizes with the α_2A adrenergic receptor and P2X₁.

In conclusion, this thesis suggests that PAR4 plays an intriguing and important role in platelets with inactived cyclooxygenase 1.  The results described in this thesis contribute to an increased knowledge of the platelet thrombin receptors.

The aim of this thesis was to determine the role of nucleoside analog activating and deactivating enzymes in nucleoside analog metabolism and resistance development. Nucleoside analogs are anti-cancer drogs and are often used to treat different leukemias, attributably to presence of high levels of nucleoside analog activating enzymes in hematopoietic cells. More recently some of the newer analogs have been used  successfully to treat solid tumors as well. We have used human leukemic cell lines, and isolated cells from patients with leukemia, to investigate the nucleoside analog activating enzymes deoxycytidine kinase (dCK) and deoxyguanosine kinase (dGK) and some of the deactivating enzymes called 5'nucleotidases (5'-NTs). We have measured mRNA expressions and enzymatic activities and correlated them with the cytotoxic response to nuc1eoside analogs and changes in cell cycle progression. We optimized and evaluated a siRNA-transfection method and decreased the activities of dCK and dGK in two different cell lines in order to find out more about their respective contribution to activation of these drogs. An expression microarray analysis of a nucleoside analog resistant cell line was also performed in order to clarify which genes are involved in development of resistance. We found that expressions and activities of dCK and dGK were not correlated. The enzyme activities of activating and deactivating enzymes changed during cell cycle progression, giving actively proliferating cells a more favorable enzymatic profile with regard to nucleoside analog cytotoxicity. The activities of dCK and dGK could be reduced transiently in leukemic and solid tumor cell lines, thereby confer either resistance or increased sensitivity to nucleoside analogs to variable degrees. Expression microarray analysis was used to evaluate the effect of the transfection method and the specificity of siRNA. We concluded that cells tolerated the transfection weIl without major effects on gene expression, and considered the siRNA used to be specific to its target. An expression microarray experiment on a nucleoside analog-induced resistant cell line revealed a hypomethylating capacity of the drog and induction of fetal hemoglobin and a multidrog resistance efflux pump as a result of the hypomethylation. This pump should not be affected by nucleoside analoges since they are not a substrate of it, and upregulation of the pump unfortunately renders the cells highly cross-resistant to different types of drogs. Our preliminary data supports our theory that it may be upregulated in order to help excrete hemoglobin that otherwise would be toxic to the cells.

Ispitani su hemijski sastav i biološka aktivnost šest samoniklih vrsta iz tribusa Scandiceae(familija Apiaceae): Anthriscus sylvestris, Anthriscus cerefolium, Chaerophyllum bulbosum,Chaerophyllum hirsutum, Chaerophyllum temulentum i Scandix pecten-veneris. LC-MS-MSanalizom ekstrakata identifikovano je više desetina sekundarnih biomolekula iz klasa flavonoida,fenilpropenskih kiselina, lignana i kumarina. GC-MS analiza pružila je uvid u sastav volatilnihkomponenti i njihov hemosistematski značaj. Potvrđeno je da sve ispitivane vrste imaju umerenoantioksidantno, antiinflamatorno i antiproliferativno dejstvo.
Chemical composition and biological activity of six wild-growing species from Scandiceae tribe (Apiaceae family) – Anthriscus sylvestris, Anthriscus cerefolium, Chaerophyllum bulbosum, Chaerophyllum hirsutum, Chaerophyllum temulentum and Scandix pecten-veneris – was examined. By LC-MS-MS analysis, a large number of secondary biomolecules was identified in extracts, including flavonoids, phenylpropenic acids, lignans and coumarins. GC-MS analysis provided insight into volatile components composition and chemosystematic significance. All investigated species exhibited moderate antioxidant, anti-inflammatory and antiproliferative activity.

Ruzmarin Rosmarinus officinalis L. (Lamiaceae) je biljka koja se u tradicionalnoj medicini na našem području koristi za postizanje analgetičkog, holeretičkog i hepatoprotektivnog delovanja. Prema Evropskoj agenciji za lekove (2010 godine), indikacije za sistemsku primenu etarskog ulja ruzmarina su lečenje dispepsije i spazama gastrointestinalnog trakta, a za spoljašnju primenu se preporučuje u lečenju umereno jakih bolova u zglobovima i mišićima i u lečenju poremećaja periferne cirkulacije. Imajući u vidu da komponente etarskog ulja ruzmarina ispoljavaju i druga, potencijalno korisna farmakološka svojstva, postoji potreba da se ova delovanja detaljnije ispitaju. Ciljevi ispitivanja su bili da se utvrdi: 1) analgetički efekat etarskog ulja ruzmarina i njegov uticaj na farmakodinamske osobine paracetamola, kodeina, diazepama i pentobarbitala kao i na farmakokinetske osobine paracetamola; 2) antioksidativni i hepatoprotektivni efekat u uslovima hemijski izazvanog oksidativnog stresa. Metodom gasne hromatografije (GC/MS i GC/FID) utvrđen je kvantitativni sastav etarskog ulja. Najzastupljenije komponente ulja koje je korišćeno u našem ispitivanju su oksidovani monoterpeni 1,8-cineol (43.77%) i kamfor (12.53%) i monoterpenski ugljovodonik α-pinen (11.51%). Suspenzija etarskog ulja ruzmarina primenjivana je miševima u dozama 10 i 20 mg/kg tm tokom sedam dana i jednokratno u farmakodinamskim testovima: test vrele ploče, test „uvijanja“ (posle intraperitonealne primene sirćetne kiseline), test za procenu motorne koordinacije životinja na rotirajućem štapu i test merenja vremena trajanja spavanja. Za ispitivanje uticaja etarskog ulja ruzmarina na farmakokinetske osobine paracetamola i za biohemijska i toksikološka ispitivanja, korišćeni su pacovi koji su tokom sedam dana tretirani suspenzijom etarskog ulja ruzmarina u dozi 5 i 10 mg/kg tm, a sedmog dana su primili paracetamol i.v. ili p.o.. Za praćenje farmakokinetskih parametara korišćeni su uzorci krvi dobijeni iz repne vene pacova u kojima su HPLC metodom merene koncentracije paracetamola, na osnovu kojih su potom određeni farmakokinetski parametri ovog leka. Antioksidativna aktivnost etarskog ulja ruzmarina je određivana in vitro (DPPH i Folin-Ciocaulteu testovima) i in vivo. Nakon žrtvovanja životinja iz prikupljenih uzoraka krvi određivani su iz seruma biohemijski parametri, pokazatelji bubrežne i jetrene funkcije, a u homogenatu tkiva jetre određivani su parametri oksidativnog stresa. Samo etarsko ulje ruzmarina ispoljava analgetičko delovanje i smanjuje visceralnu bol izazvanu sirćetnom kiselinom. Pored toga, potencira analgetički efekat kodeina i paracetamola. Etarsko ulje ruzmarina značajno smanjuje hipnotičko delovanje pentobarbitala i sprečava poremećaj motorne koordinacije nakon primene diazepama. Etarsko ulje ruzmarina ne utiče značajnije na oralnu biološku raspoloživost paracetamola. Višekratna primena različitih doza etarskog ulja ruzmarina ne izaziva toksične promene u krvi i jetri ispitivanih životinja. Primena etarskog ulja ruzmarina štiti životinje od reaktivnih kiseoničnih vrsta, umanjuje posledice izloženosti oksidativnom stresu i ispoljava značajno hepatoprotektivno delovanje.
Rosemary Rosmarinus officinalis, L.(Lamiaceae) is traditionally used in folk medicine for its analgetic, choleretic and hepatoprotective properties. According to the recommendation of European Medicines Agency from 2010, rosemary essential oil can be used for treating dyspepsia and mild spasmodic disorders of the gastrointestinal tract, and also externally as an adjuvant in the relief of minor muscular and articular pain and minor peripheral circulatory disorders. Different studies conducted with rosemary essential oil show other pharmacological effects of main components of the oil. The aim of this study was to examine: 1) analgetic effects of rosemary essential oil and its influence on the pharmacodynamic properties of paracetamol, codeine, diazepam and pentobarbital, and also its influence on the pharmacokinetic properties of paracetamol; 2) antioxidant and hepatoprotective effects on the parameters of chemicaly induced oxidative stress. The quantification of chemical constituents of the essential oil was carried out by gas chromatography (GC/FID and GC/MS). The major compounds that were identified and quantitated by GC-FID and GC-MS were oxygenated monoterpens 1,8-cineole (43.77%), camphor (12.53%) and monoterpene hydrocarbon α-pinene (11.51%). The suspension of rosemary essential oil was applied to mice orally (doses: 10 and 20 mg/kg b.w.) for seven days and in single dose for the pharmacodynamic tests: hot plate, writhing, rotharod and sleeping time. Rats treated with suspension of rosemary essential oil for seven days orally (doses: 5 and 10 mg/kg b.w.) were used for the examination of influence of essential oil on the pharmacokinetic properties of paracetamol. Then on the 7th day the paracetamol was applied to them p.o. or i.v.. The parameters of pharmacokinetic were analyzed in blood samples obtained from rats tail veins. The HPLC method was used for measurement of concentration of paracetamol in blood samples. Those concentrations were used for calculation of the pharmacokinetic parameters. The antioxidant activity of the rosemary essential oil was evaluated in vitro (with DPPH and Folin-Ciocaulteu tests) and in vivo. The animals were sacrificed and the samples of blood and liver were taken. The obtained serum was used for determination of standard biochemical parameters and the parameters of oxidative stress were analyzed in obtained liver homogenates. The essential oil of rosemary shows analgetic properties and it decreases visceral pain induced with intraperitoneally injected acetic acid. The rosemary essential oil increases pharmacological effects of codeine and paracetamol. Also, this oil reduces pentobarbital-induced sleeping time and diminishes diazepam-induced disorder of psychomotor coordination. The essential oil of rosemary does not change paracetamol bioavailability. The rosemary essential oil applied in multiple doses does not induce toxic changes in blood and liver samples obtained from animals. The use of rosemary essential oil protects animals from reactive oxygen species, decreases the effects caused by oxidative stress and shows significant hepatoprotective effect.

The neuropeptide Y (NPY) family of structurally related peptides includes NPY, peptide YY (PYY) and pancreatic polypeptide (PP). They bind to G-protein coupled receptors named Y receptors, and include in mammals Y1, Y2, Y4, Y5, Y6 and in non-mammalian vertebrates also Y7, Yb and Yc. Subtypes Y1 and Y5 stimulate appetite, while Y2 and Y4 have the opposite effect in mammals. The studies described here concern human Y1 and Y4, chicken Y6 and Y7, and zebrafish Y2.

Site-directed mutagenesis of human Y1 identified sites important for binding of NPY and PYY as well as Y1 antagonists. The results clarify contradictory findings previously reported by others and identify new sites of interaction. A three-dimensional structural model of the Y1 receptor based upon the high-resolution structure of bovine rhodopsin was generated that increases our understanding of ligand-receptor interactions and hopefully will facilitate the design of novel subtype-selective agonists and antagonists.

Two naturally occurring variants of human Y4 with substitutions R240C and V276M have been found in a sample of obese children. Functional studies in vitro showed that the cellular response to PP was greatly decreased for R240C and may provide a causative link to juvenile obesity.

The genes for chicken Y6 and Y7 were found to be located ~1 megabase apart on chromosome 13 syntenic to the human Y6 pseudogene. Y6 mRNA has widespread expression whereas Y7 mRNA was found only in adrenal gland. Truncated fragments of PYY had lower affinity to chicken Y7 and zebrafish Y2 than to Y2 from mammals and chicken. The results suggest that Y2 in mammals acquired the ability to bind truncated PYY, i. e. PYY_3-36, fairly recently, which has implications for its role in appetite inhibition.

These differences between receptor subtypes in sequences and pharmacological properties will be useful to further elucidate the structure and activation of Y receptors by site-directed mutagenesis.

Botulinumtoxin: För och emot dess användning i skönhetsbranschen Bakgrund: Botulinumtoxin (BTX) är ett kraftfullt neurotoxin som produceras av den grampositiva och anaeroba bakterien Clostridium botulinum. BTX används i medicinskt syfte vid behandling av till exempel kronisk migrän, spasticitet efter stroke, och urinläckage vid nervskada. Utöver det används BTX inom skönhetsbranschens som då injiceras i mycket låga doser under huden i muskler för att minska rynkor i olika ansiktsregioner Syfte: Användningen och lämpligheten inom skönhetsbranschen har ifrågasatts , och syftet med detta arbeta har därför varit att utvärdera och ta ställning till om BTX fortsättningsvis bör användas i kosmetiskt syfte. Metod: Litteraturstudie där vetenskapliga artiklar söktes i databasen PubMed med kombinationer av sökord, artiklar som sedan kunde användas för att besvara den aktuella frågeställningen. Resultat: Olika studier har visats att BTX är ett effektivt och säkert toxin vid skönhetsbehandling, men flera studier visade också att BTX-relaterade biverkningar uppkom efter skönhetsbehandling. Slutsats: BTX bör inte fortsätta användas för skönhetsbehandling trots positiva effekter mot rynkor i ansiktet som visades i studierna. Injektion med BTX kan bidra till att kunder i skönhetssalonger blir svårbedömda patienter vid besök hos läkare i primärvården som inte förstår att de problem patienten söker för beror på BTX-inducerade biverkningar efter skönhetsbehandling med BTX.

Background: Contraceptives for men has existed for centuries. However, the only contraceptives available for men are condoms, withdrawal during sexual intercourse or undergoing Vas occlusion. The issues currently with these methods are that many couples are not comfortable with using condoms and vasectomy is a non-reversible contraceptive as it requires surgery. Currently, only research has been made with regards to hormonal contraceptives for men. The basis for hormonal contraceptives for men is that it disturbs hypothalamic-pituitary–gonadal axis, by suppressing GnRH, LH and FSH in order to suppress spermatogenesis. The substances that have been clinically tested as a potential hormonal contraceptive for men are androgens, androgens together progestins and synthetic androgens. Aim: The aim of this thesis was to examine the effectiveness of the substances that have been used in past and recent clinical trials and the adverse drug reactions/effects. Method: Results from the clinical trials were collected through PubMed with regards to preordained criteria, which resulted in 18 scientific articles being used in the results. Results: Overall, the results showed that the majority of substances used in previous and recent clinical trials are effective in suppressing spermatogenesis. The results also showed that a majority of substances used in previous and recent clinical trials had severe adverse drug effects severe amongst the participants. Discussion and Conclusion:  In summary, the research shows that amongst the substances used in recent and current clinical trials, that synthetic androgens have the best potential as a hormonal contraceptive for men with regards to effectiveness and adverse drug effects.

Depression är den näst mest kostsamma sjukdomen för samhället efter hjärt-kärlsjukdom, främst på grund av långa sjukskrivningsperioder. Sjukdomen kan uppstå när som helst från sex månaders ålder, men prevalensen ökar med åldern. Det finns ett antal stressrelaterade faktorer som skulle kunna leda till depression, så som stor sorg, verbala eller fysiska övergrepp samt en svår barndom. Vad som orsakar sjukdomen är ännu inte helt känt, men det finns teorier att halterna av serotonin och noradrenalin är lägre hos deprimerade personer. Behandling som används är olika former av samtalsterapi, men även läkemedel så som selektiva serotoninåterupptagshämmare (SSRI). Det finns teorier som sammankopplar användandet av SSRI med självmord, framförallt hos personer ≤19 år. Syftet med detta litteraturarbete var att undersöka om SSRI preparat har någon effekt på depression hos barn och ungdomar och om de är säkra eller kan få allvarliga konsekvenser så som självmord. Sökningar i PubMed gjordes för att hitta relevanta artiklar. Fem av de åtta inkluderade studierna rapporterade olika effekter och säkerhet hos olika SSRI preparat bland barn och ungdomar, jämfört med placebo. Två andra studier undersökte förekomsten av suicidalitet till följd av läkemedlen. Den sista studien jämförde toxikologiska data från Rättsmedicinalverket med receptregistret på antidepressiva läkemedel från  Socialstyrelsen. Endast två av de fem studerade preparaten (fluoxetin och citalopram) hade en bättre effekt än placebo i hela populationen och ytterligare ett (sertralin) hade bättre effekt hos ungdomar. Det begicks inga självmord i studierna.    De studier som har granskats i detta arbete tyder på att olika SSRI preparat har olika bra effekt samt olika säkerhetsprofiler. Det sågs inget tydligt samband mellan behandlingen och självmord, men en något förhöjd risk för suicidalitet.

Mental disorders, like anxiety and depression, are a major health problem and can appear early in life. Developing new compounds for anxiety and other mental disorders is desirable and requires good animal models. Two widely used tests to evaluate fear and anxious behavior in rodents are the open field (OF) and elevated plus maze (EPM) tests. The models are simple and it has been argued that different environments give different behavior profiles and measure different kind of behavior. The multivariate concentric square fieldTM (MCSF) is a new test that gives the rodent opportunity to visit different environments and to evaluate more parameters in one test. Validation of the tests is mainly done with adult animals. The aim with this study is to study adolescent rat behavior in the OF, OF with start box, EPM and MCSF tests and to see if there is any difference in running trials in the morning or in the afternoon. A total of 48 adolescent male Wistar rats were divided into 12 rats per group. The groups were OF and MCSF, OF with start box and MCSF, EPM and MCSF, and repeated testing in MCSF. Each session was 20 minutes and there was one week between the two tests. Three rats where run in the morning and three in the afternoon. Statistical analysis did show significant difference in some parameters in the comparison between running trials in the morning or in the afternoon. Repeated testing in the MCSF yielded differences in trial two compared to trial one. No significant difference was found in the trend analysis. The results show individual differences and with larger groups results may have been more liable. In this study some differences were found between morning and afternoon groups. Lots of data have been generated and there are many opportunities to use the same data for additional analyses.

Epilepsi är ingen speciell sjukdom utan ett symtom på en hjärnskada eller störd nervcellsfunktion i hjärnan. Epileptiska anfall beror på abnorm urladdning i hjärnans nervceller. Idag lever omkring 60 000 d.v.s. 0,5-1 % av Sveriges befolkning med epilepsi. Risken att drabbas är störst under det första levnadsåret och efter 65-årsålder då risken att drabbas av stroke är som störst. Behandling av epilepsi används i syfte att hindra uppkomst av anfall och göra det möjligt för den drabbade att leva ett relativt normalt liv. Antiepileptika dämpar aktiviteten i hjärnan och reducerar därmed risken för anfall. Under flera år har man försökt utveckla nya antiepileptika mot andra möjliga targets än de som finns idag, bl.a. GABA-transporthämmare. Det enda förekommande läkemedlet med GABA transporthämmande effekt är tiagabin men detta är inte registrerat som läkemedel i Sverige. Syftet med denna studie var att undersöka om GABA-transporthämmare skulle kunna användas som läkemedel mot epilepsi. Metoden som användes var en litteraturstudie där vetenskapliga artiklar hämtades från PubMed, ELIN, Cochrane och Google Scholar. Arbetet baseras på 4 experimentella originalartiklar och en metaanalys. Artiklarna beskriver antiepileptiska effekter och/eller relaterade egenskaper för olika substanser med hämmande effekter på olika GABA- transportörer. Dessa hämmare, ensamma eller i kombination, visades ge kramplösande effekt i olika djurmodeller av epilepsi. Hämmare av olika GABA-transportörer, till exempel tiagabin och EF1502, gav synergistisk effekt, medan hämmare av samma GABA-transportör, till exempel tiagabin och LU-32-176B, resulterade i additiv effekt. Hämning av olika GABA-transportörer i olika celltyper i och runt synapsklyftan verkar därför kunna ge synergistisk effekt. Ingen synergistisk effekt observerades för toxiska effekter. Det finns anledning att tro att ytterligare läkemedel med effekter på GABA-transportörer kan komma att finnas i framtiden för behandling av epilepsi.

Epilepsy is not a specific disease but a symptom of brain injury or impaired nerve cell function in the brain. Epileptic seizures are symptoms of abnormal activity in the brain neurons. Today, about 60 000 i.e. 0.5-1% of the Swedish population live with epilepsy. The risk of being affected is greatest during the first year of life and after the age of 65 years when the risk for stroke is greatest. The treatment of epilepsy is used in order to prevent the onset of seizures and to allow the patient to live a relatively normal life. Anticonvulsants dampen the activity in the brain and thus reduce the risk of seizures.

During many years, attempts have been made to develop new anticonvulsants against other potential targets than those that exist today, for example GABA-transporter inhibitors. The only presently used medicine with GABA-transporter inhibiting effect is tiagabine, but this is not licensed as a pharmaceutical drug in Sweden.

The aim of this study was to investigate whether GABA-transport inhibitors could be used as medication for epilepsy. The method that was used was a literature study in which scientific articles were chosen from PubMed, ELIN, Cochrane and Google Scholar. The work is based on 4 original research articles and one meta-analysis. The articles describe antiepileptic effects and/or related properties of various substances with inhibitory actions on different GABA-transporters. These inhibitors, alone or in combination, were shown to have anticonvulsant effects in several different animal models of epilepsy. Inhibitors of different GABA transporters, such as tiagabine and EF1502, resulted in synergistic effects, while inhibitors of the same GABA transporter, such as tiagabine and LU-32-176 B, resulted in additive effects. Inhibition of various GABA transporters in different cell types in and around synapses therefore seems to provide synergistic effects. No synergistic effect was observed for toxic effects. There is reason to believe that additional drugs with effects on GABA transporters may be used in the future for the treatment of epilepsy.

J1 (L-melphalanyl-L-p-fluorophenylalanyl ethyl ester) is a dipeptide derivative of the alkylating agent melphalan with increased cytotoxicity. In this thesis the preclinical pharmacology of J1 has been characterized.

Our results show that J1 rapidly enters the cells, where melphalan is released by hydrolysis. The maximum concentration (C_max) of melphalan was detected 15 min after exposure to J1 in human cancer cell lines. In comparison, melphalan exposure resulted in a 10-fold lower C_max that was shifted to later time points. J1 induced more DNA damage and apoptosis than melphalan. The cytotoxic activity and release of melphalan from J1 were inhibited by preincubating cells with the aminopeptidase inhibitor bestatin. In accordance with these results, we showed that J1 is a substrate for aminopeptidase N (APN), which may result in increased tumor selectivity.

J1 effectively inhibited cell growth in a set of neuroblastoma cell lines. Athymic mice carrying neuroblastoma xenografts were treated either with equimolar doses of melphalan or J1. J1 inhibited the tumor growth more effectively than melphalan and the untreated control, and was associated with higher caspase-3 activation, fewer proliferating tumor cells and decreased mean vascular density.

J1 and melphalan showed similar activity profiles when tested in 176 primary tumor cell cultures from patients, but J1 exhibited 50- to 100-fold higher potency. The difference was greater in some diagnoses (e.g. breast cancer, NHL and AML), and was exceptionally large in some breast cancer samples with aggressive phenotypes. A combination screening of J1 and standard chemotherapeutics yielded mostly additive interactions, except for etoposide which induced synergy in all tested cell lines.

In conclusion, the melphalan prodrug J1 is effectively transported into the cells, where aminopeptidases (for example APN) catalyze the formation of melphalan. J1 shows promising preclinical potential in the diagnoses neuroblastoma and breast cancer.

Cancer is a common disease and due to problems with resistance against cancer drugs and the limited benefit from chemotherapy in many diagnoses, there is a need to develop new cancer drugs. In this thesis new methods to screen for cancer drugs and to evaluate their mechanism of action are discussed.

In Paper I, it was found that by studying the gene expression of a cell line panel and combining the data with sensitivity data of a number of cytotoxic drugs, it was possible to cluster compounds according to mechanism of action as well as identifying genes associated with chemosensitivity.

In Paper II, studies of compounds with selective activity in drug-resistant cell lines revealed the glucocorticoids as a group of interesting compounds. The glucocorticoid receptor was overexpressed in 8226/Dox40 and the difference in sensitivity was abolished when the cells were treated with a glucocorticoid receptor antagonist.

In Paper III, an image-based screening method for new proteasome inhibitors was successfully developed and the compounds disulfiram, PDTC and NSC 95397 were identified as inhibitors of the proteasome.

In Paper IV, disulfiram and PDTC were shown to induce cytotoxic activity, to inhibit the activation of the transcription factor NFkappaB and to inhibit the degradation of proteins normally degraded by the proteasome.

In Paper V, NSC 95397 was shown to be cytotoxic to all cells in the resistance-based cell line panel as well as to patient samples from a variety of cancer diagnoses. Connectivity Map was successfully used as a tool to propose a new mechanism of action of NSC 95397. The gene expression induced by NSC 95397-treatment was similar to that induced by several proteasome inhibitors not present in the Connectivity Map.

Bakgrund: Alzheimers sjukdom (AD) är neurodegenerativ och kännetecknas framför allt av det tidiga symtomet minnesförlust. De patologiska skadorna i hjärnan utmärks av senila plack och neurofibrillära nystan (NFTs). Senila plack består av peptiden amyloid-b (Ab) och NFTs innehåller proteinet tau. Ab har länge stått i fokus när det kommer till patogenesen vid AD och forskning kring behandlingsstrategier mot sjukdomen har länge riktat sig mot denna peptid, men än så länge utan lyckade resultat. Mer fokus har riktats till tau och i nuläget finns det ett antal tau-riktade behandlingsstrategier under utveckling och tau-immunoterapi är en av dessa. Syfte: Idag finns det inte någon behandling som botar AD vilket betyder att det är viktigt att hitta nya behandlingsstrategier mot sjukdomen. Tidigare studier tyder på att tau är ett potentiellt mål för behandling mot AD och syftet med detta arbete var därför att undersöka om tau-immunoterapi är en möjlig behandlingsstrategi vid AD. Metod: Sekundärdata samlades in från databasen PubMed. Datan granskades och bedömdes som användbar eller inte. Detta resulterade i att sju orginalartiklar valdes till arbetet, sex studier i djurmodeller och en klinisk studie.  Resultat: Studierna i djurmodellerna visade att tau-immunoterapi reducerade patologiskt tau och i hälften av studierna observerades även en förbättring av de kliniska symtomen. Den kliniska studien visade att AADvac1 är säkert och ansågs vara en lovande kandidat för vidare studier. Slutsats: Resultaten från studierna visar att tau-immunoterapi är en lovande behandlingsstrategi vid AD. Trots detta krävs det fler kliniska studier som undersöker tau-immunoterapi för att säkerställa om behandlingen har någon klinisk effekt på patienter med AD eller inte.

The period of early life is important for the development of individual brain function and behavior. Human studies have shown altered vulnerability to develop psychopathology and/or excessive drug intake, possibly leading to dependence, as a consequence of early life experiences. In the present thesis, maternal separation (MS), an experimental model for studies of early environmental influences, was used to investigate long-term effects on neurochemistry, voluntary ethanol intake and exploration and risk assessment behavior in rats. Rat pups were assigned to one of three different rearing conditions: daily 15 min (MS15) or 360 min (MS360) of MS and normal animal facility rearing (AFR) during the first three weeks of life. Measurements of adult endogenous opioid peptide levels, opioid- and dopamine receptor density revealed minor MS-induced effects on the opioid system whereas interesting alterations were found in dopamine receptor density. Long-term effects on voluntary ethanol intake showed distinct MS-induced alterations in male Wistar and ethanol-preferring AA (Alko, Alcohol) rats. Female Wistar rats were unaffected, indicating sex differences in the effects of MS on ethanol intake. Male MS15 rats generally had a slower acquisition phase and a low subsequent ethanol intake whereas male MS360 rats had a high ethanol intake. MS15 is therefore suggested to protect against a high voluntary ethanol intake in male rats whereas MS360 may serve as a risk factor. The recently established concentric square field test indicated alterations in risk assessment as well as an increased exploratory drive and somewhat higher risk-taking behavior in adult MS360 rats, while minor effects were seen in MS15 rats. Altogether, these results demonstrate that environmental influences during the period of early life can have long-term effects on neurochemistry and behavior. Of special interest is the finding that MS altered the inherited high ethanol intake in adult ethanol-preferring AA rats.

Antalet äldre har ökat i hela världen och fortsätter att öka. I takt med stigande ålder ökar risken för sjukdomar och därmed även läkemedelsanvändningen. Personer över 80 år konsumerar i genomsnitt 5,8 läkemedel per person och äldre på sjukhem tio läkemedel per person vilket är en ökning med 60 % sedan slutet av 1980-talet. Anledningen till denna ökning är att det idag finns stora möjligheter att förebygga och behandla många sjukdomar eftersom det hela tiden utvecklas nya läkemedel och behandlingsmetoder. Polyfarmaci och olämplig förskrivning av läkemedel till äldre över 65 år enligt Beers kriterier (se bilaga I) är ett växande och världsomspännande hälsoproblem. Vid polyfarmaci används i genomsnitt fem eller flera olika läkemedel. Polyfarmaci ökar i takt med stigande ålder och är vanligast hos kvinnor samt hos lågutbildade individer. Med ökat antal läkemedel ökar också risken för biverkningar samt interaktioner eftersom äldre är känsligare på grund av åldersförändringar och sjukdom. Syftet med studien var att belysa förekomsten av polyfarmaci och olämplig förskrivning av läkemedel till äldre samt dess konsekvenser. Studien utfördes som en litteraturstudie där 17 vetenskapliga artiklar analyserades. Resultatet visar att en tredjedel av de äldre patienterna konsumerar olämpliga läkemedel, enligt Beers kriterier, vilket leder till onödigt lidande och ökad sjukhusvistelse. Det är därför viktigt att all vårdpersonal är väl insatta i de åldersförändringarna som sker hos den äldre individen samt har god kunskap inom farmakologi. Sjuksköterskor och läkare bör även förbättra samarbetet med farmaceuterna för att öka patientsäkerheten.

 

 

The elderly population is increasing all over the world. Aging is associated with diseases resulting in increased medical consumption. Elderly over 80 years consume in average 5,8 different drugs. Nursing home residents consume ten drugs which represents an increase of 60 % within the last two decades. This development is based on the increasing progress within the field medical treatment. Polypharmacy and inappropriate prescribing in elderly over 65 years, according to Beer´s criteria (annex I), results in a growing and worldwide health problem. Polypharmacy comprises use of multiple drugs (mostly five or more per day). Polypharmacy is associated with increased age and is most common in women and low educated individuals. Multiple medications increase the risk of adverse drug reactions and drug-drug interactions especially in old and frail persons with comorbidity. The aim of the study was to elucidate the prevalence and the consequences of polypharmacy and inappropriate prescribing in elderly. The study was based on a literature study in which 17 articles were analyzed. The result shows that one third of the elderly patients consume inappropriate medications, according to Beer´s criteria, which are associated with unnecessary suffering and increased hospital admission. It´s important that health care personnel gains understanding about the pharmacological consequences of body composition changes in older adults. Nurses and physicians should also improve their cooperation with pharmacists to increase knowledge leading to better patient safety.

The superfamily of G protein-coupled receptors (GPCRs) is one of the largest protein families of mammalian genomes and can be divided into five main families; Glutamate, Rhodopsin, Adhesion, Frizzled, and Secretin. GPCRs participate in most major physiological functions, contributing to the fact that they are important targets in drug discovery. In paper I we mined the human and mouse genomes for new Adhesion GPCR genes. We found two new human genes (GPR133 and GPR144) and 17 mouse Adhesion genes, bringing the number up to 33 human and 31 mouse genes. In paper II we describe 53 new splice variants for human Adhesion receptors supported by expressed sequence tags (EST) data. 29 of these variants seem to code for functional proteins, several of which lack one or more functional domains in the N-termini. Lack of certain domains is likely to affect ligand binding or interaction with other proteins. Paper III describes the Glutamate GPCR in human, mouse, Fugu, and zebrafish. We gathered a total of 22 human, 79 mouse, 30 Fugu, and 32 zebrafish sequences and grouped these into eight clans using phylogenetic methods. The report provides an overview of the expansion or deletions among the different branches of the Glutamate receptor family. Paper IV focuses on the trace amine (TA) clan of Rhodopsin GPCRs. We identified 18 new rodent genes, 57 zebrafish genes, and eight Fugu genes belonging to the clan. Chromosomal mapping together with phylogenetic relationships suggests that the family arose through several mechanisms involving tetraploidisation, block duplications, and local duplication events. Paper V provides a comprehensive dataset of the GPCR superfamily of human and mouse containing 495 mouse and 400 human non-olfactory GPCRs. Phylogenetic analyses showed that 329 of the receptors are found in one-to-one orthologous pairs, whereas other receptors may have originated from species-specific expansions.

The sequencing of genomes has caused a growing demand for functional analysis of gene products. This research field named proteomics is derived from the term proteome, which by analogy to genome is defined as all proteins expressed by a cell or a tissue. Proteomics is however methodologically restricted to the analysis of proteins with higher molecular weights. The development of a technology which includes peptides with low molecular weight and small proteins is needed, since peptides play a central role in many biological processes.

To study endogenous peptides and hormones, the peptidome, an improved method comprising rapid deactivation in combination with nano-flow liquid chromatography (LC) and mass spectrometry (MS) was developed. The method has been used to investigate endogenous peptides in brains of mouse and rat. Several novel peptides have been discovered together with known neuropeptides.

To elucidate the post mortem time influence on peptides and proteins, a time course study was performed using peptidomics and proteomics technologies. Already after three minutes a substantial amount of protein fragments emerged in the peptidomics study and some endogenous peptides were drastically reduced with increasing post mortem time. Of about 1500 proteins investigated, 53 were found to be significantly changed at 10 minutes post mortem as compared to control. Moreover, using western blot the level of MAPK phosphorylation was shown to decrease by 95% in the 10 minutes post mortem sample.

A database, SwePep (a repository of endogenous peptides, hormones and small proteins), was constructed to facilitate identification using MS. The database also contains additional information concerning the peptides such as physical properties. A method for analysis of LC-MS data, including scanning for, and further profiling of, biologically significant peptides was developed. We show that peptides present in different amounts in groups of samples can be automatically detected.

The peptidome approach was used to investigate levels of peptides in two animal models of Parkinson’s disease. PEP-19, was found to be significantly decreased in the striatum of MPTP lesioned parkinsonian mice. The localization and expression was further investigated by imaging MALDI MS and by in situ hybridization. The brain peptidome of reserpine treated mice was investigated and displayed a number of significantly altered peptides. This thesis demonstrates that the peptidomics approach allows for the study of complex biochemical processes.

Obesity is a complex and a highly individualized disease and the molecular mechanisms behind this disorder need to be better elucidated. Identification of genes and genetic variants that are involved provide opportunities to establish a genetic understanding of the disease. These findings may also provide more rational approaches to therapy, either by identifying underlying causes or point out the need for different treatments. In addition, the timing and severity of obesity may provide insights into the aetiology of obesity and also identify age-specific determinants of weight gain. Recently, genome-wide association studies have led to a rapid progress in our understanding of the genetic basis of various diseases and candidate genes for obesity have been identified. The overall aim of this thesis was to investigate the genetic impact on severity of childhood obesity and the associations between obesity and genetic variants in the fat mass and obesity associated gene, FTO, and MGAT1, the gene encoding mannosyl (α-1,3-)-glycoprotein β-1,2-N-acetyl-glucosaminyltransferase. We show that the impact of parental body mass index (BMI) on the severity of obesity in children is strengthened as the child grows older, whereas the age at obesity onset is of limited importance. By association studies, we show that single nucleotide polymorphisms downstream MGAT1 influence susceptibility to obesity. Moreover, these variants affect the levels of unsaturated fatty acids and desaturase indices, variables previously shown to correlate with obesity. Furthermore, one variant in the first intronic region of FTO is associated with obesity among children but not with BMI or other measures of adiposity at older ages. However, this variant shows a weight-dependent association with cognitive function among elderly men. By direct sequencing, we identified novel variants in FTO, affecting glucose homeostasis in a BMI-independent manner. Furthermore, we found gender specific effects for FTO, both regarding obesity susceptibility and related phenotypes.

Abstrakt Bakgrund: Alzheimers sjukdom (AD) är en av de vanligaste neurodegenerativa sjukdomarna och anses vara den främsta orsaken till nedsatt eller försämring av minnet hos äldre. De vanligaste symptomen på AD är förlust av intellektuell förmåga som så småningom resulterar i dysfunktion i det dagliga livet. Forskningar har visat att amyloida β-peptider kan ha en avgörande roll i den patologiska processen för neurodegenerativ sjukdom inklusive AD. Amyloid β-peptider är en nyckelmolekyl vid AD där en ansamling av detta peptid ansågs initiera den patologiska kaskaden av AD, inklusive bildandet av senila plack och neurofibrillära trassel vilket leder till neuronal förlust och demens. Med tanke på att AD patienter uppvisar ökad nivå av Aβ40-42 i både blodet och cerebrospinalvätskan, anses detta vara en biomarkör för sjukdomen. Därav används dessa biomarkörer för prognos och behandling av Alzheimer sjukdom. β-sekretashämmare blockerar den första hastighetsbegränsande proteolytiska klyvningen av amyloida prekursorpeptider. Detta bör resultera i produktionsreducering av aggregerande Aβ-peptider. Forskning pågår kring användandet av β-sekretashämmare för behandling av AD och dess kliniska relevans. Syfte: Syftet med detta fördjupningsprojekt är att undersöka om β-sekretashämmare påverkar Aβ-koncentrationen i cerebrospinalvätska och plasma. Samt att diskutera ifall reduceringen medför en inbromsning av de degenerativa processer som AD orsakar. Metod: Metoden som användes är en systematisk litteraturöversikt där fem studier granskades. Resultat: Baserat på de vetenskapliga originalartiklarna som använts har β-sekretashämmare minskat koncentrationen på β-amyloid i både cerebrospinalvätska och plasma med mer än 50 % hos alla män och kvinnor mellan 18–75 år. Slutsats: β-sekretashämmare reducerar Aβ-nivåerna. Flera kliniska studier behövs för att bekräfta effekten av denna minskning för att se om den har en klinisk betydelse.

Bipolar disorder is a severe, affective illness which causes a person to alternate between episodes of pathologically elevated mood (mania) and depression – in between these episodes the mood is normal. The length and intervals between the episodes are highly variable among the ill, as are the severity which is divided into two major subgroups: bipolar I and II. Bipolar I disorder is the most severe because of the patient’s inability to realize that he or she is currently in a manic phase, in the case of bipolar II disorder, the patient is aware that he or she has an abnormally raised mood, a state which is called hypomanic state instead of manic. The cause of bipolar disorder is yet to be discovered, although both genetic and environmental aspects probably weigh in. Bipolar disorder is treated with a combination of psychotherapy and pharmacology, the most common and oldest treatment regimen being that of lithium, an alkali metal which has been used as a mood stabilizer for over a century. Although being proven effective, lithium can cause thyroid malfunction and has a complicated renal elimination process that can be affected by diet and heritage. This makes regular blood testing for determining concentration of lithium necessary. Lithium can also be toxic in relatively low doses. Lamotrigine is an anticonvulsive drug used for treating epilepsy, but has been proven to also have mood stabilizing properties. To find out if lamotrigine can be used as a treatment for bipolar disorder, both as maintenance treatment as well as acute treatment in manic/hypomanic/depressive episodes, papers from clinical trials where lamotrigine has been tested with lithium and/or placebo have been read. The results show that lamotrigine is as effective as lithium in maintenance and acute treatment of depression, although some papers show conflicting results on the effectiveness compared to placebo. The conclusion is that lamotrigine seems to be a promising drug that can be helpful in the treatment of bipolar disorder.

Fler än 350 miljoner människor i världen hade en depression 2012 och sjukdomen tillhör tillsammans med hjärt-kärlsjukdom de mest kostsamma sjukdomarna i västvärlden. 2007 behandlades över 700 000 personer i Sverige med antidepressiva läkemedel till en kostnad av 990 miljoner kronor. Depression kan vara unipolär (endast nedstämdhet) eller bipolär (även med maniska inslag). En primär ”egentlig” depression, major depression, MDD, har ofta en biologisk orsak. Bland annat har man funnit att halterna av signalsubstanserna serotonin och noradrenalin är lägre hos deprimerade. Behandling av depression idag inkluderar antidepressiva läkemedel, elektrokonvulsiv behandling (eng. electroconvulsive therapy (ECT)) och psykoterapi. Syftet med denna litteraturstudie var att jämföra venlafaxin, som är en selektiv serotonin- och noradrenalinåterupptagshämmare (SNRI), med traditionella selektiva serotoninåterupptagshämmare (SSRI) i effekt och säkerhet vid behandling av unipolär depression hos vuxna.  Sju studier, som hämtades från Pubmed och Medline, granskades i detta arbete. De utgjordes av två meta-analyser och fem randomiserade kliniska prövningar. Resultatet visade att i fyra studier hittades ingen statistiskt signifikant skillnad i respons mellan de patienter som fick venlafaxin och de som fick SSRI. I de tre studier som uppvisade en statistiskt signifikant skillnad var venlafaxin effektivare än SSRI. I studie 2 hittades skillnaden i en subgrupp med de sjukaste patienterna (p=0.0121),  i studie 6: RR=1.06, 95% KI 1.01-1.12 och i studie 7: OR=1.15, 95% KI 1.02-1.29.  Endast en av studierna påvisade en statistiskt signifikant skillnad i remission, studie 7, och även här till fördel för venlafaxin  (OR=1.19, 95% KI 1.06-1.34). Dock orsakade venlafaxin fler bortfall p.g.a. biverkningar än SSRI i sex av studierna. Sammanfattningsvis kan venlafaxin anses något effektivare än SSRI, men detta har troligtvis ingen stor klinisk betydelse förutom eventuellt vid behandling av svårare depressioner. Biverkningarna bör vägas in i valet av behandling. Det vore intressant att se fler studier som jämför venlafaxin med SSRI vid svårare depressioner.

Neuroblastoma, a neuroendocrine tumor, is the most common cancer in infancy. 75 % of those affected are under the age of 5. The disease is heterogeneous and survival rate is low.   Current treatment of neuroblastoma consists of surgery, radiation and chemotherapy, where the targets for the treatment are the malign cells. Due to the cancer cells instable genome there is a risk for resistance development. This negatively impacts the treatments goal of hindering tumor growth and spread.  Tumor growth is not only determined by malign cells but also the interactions of those tumor cells with tumor vessels and different types of cells in the tumor stroma.   The aim of this paper is to develop a relevant histological method to study the properties of tumor stroma in tumor sections retrieved from human NB tumor xenografts in mice treated with angiogenesis inhibitors SU11657 and Zoledronic acid. The study is a continuation of previous studies with the inhibitors which have shown good effect on tumor growth and angiogenesis on neuroblastoma.   In the short term treatment with SU11657 and Zoledron acid showed that tumor growth declined. In the longer treatment with SU11657 the growth didn’t decline with the same rate compared to the short term treatment. Angiogenesis on the other hand decreased in all the treatments independent of treatment duration. The histological staining with Sirius red revealed that treated tumors had an increased amount of stroma compared to the untreated tumors.   In conclusion the relative increase of tumor volume, decreased number of vessels and expansion of tumor stroma in the longer treatment with SU11657 indicated that tumors might survive the angiogenesis inhibitor treatment through expansion/activation of its stroma. The histological staining with Sirius red in saturated picric acid marked the collagen, i.e. stroma, well and enabled quantification of the stroma.

The practise of microdosing certain psychedelic drugs, that is consuming doses below thethreshold of experiencing psychedelic side effects, is increasing in popularity. Microdosing ismainly practised to reach the positive side effects of psychedelic drugs, mainly for themood-enhancing and antidepressant effects without having to experience the psychedelic sideof the drug. In this systematic literature review we searched the literature to explore howmicrodosing can be used as an alternative treatment for treatment of depression. While theliterature indicates that microdosing psychedelics could be useful in treatment of depression,there are also clear limitations in the published research. The main limitations include bias,uneven distribution of gender, excludment of participants with a history of mental illnesses,heterogeneity of dosing schedules. This results in contradictory findings. Experimentalstudies show no effects on depression, whereas qualitative and observational studies showpositive effects regarding depression and positive lifestyle changes. This review provides anoverview of the field today and provides a useful base for future studies that are necessary tofully answer whether microdosing is a useful way to treat depression or not.
Övningen av mikrodosering av vissa psykedeliska droger vilket är att ta doser under tröskelnför att uppleva psykedeliska effekter ökar i popularitet. Mikrodosering praktiseras för att nåde positiva biverkningarna av psykedeliska drogerna främst för de humörhöjande ochantidepressiva effekterna utan att behöva uppleva den psykedeliska sidan av läkemedlet. Idenna systematiska litteraturöversikt sökte vi i litteraturen för att undersöka hurmikrodosering kan användas som en alternativ behandling för behandling av depression.Medan litteraturen indikerar att mikrodosering av psykedelika kan vara användbart vidbehandling av depression finns det också tydliga begränsningar i den publiceradeforskningen. De främsta begränsningarna inkluderar partiskhet, ojämn könsfördelning,uteslutning av deltagare med tidigare psykiska sjukdomar och heterogenitet i doseringsscheman. Detta resulterar i motsägelsefulla fynd. Experimentella studier visar inga effekter pådepression, medan kvalitativa och observationsstudier visar positiva effekter när det gällerdepression och positiva livsstilsförändringar. Denna översyn ger en översikt över områdetidag och ger en användbar grund för framtida studier som är nödvändiga för att helt svara påom mikrodosering är ett användbart sätt att behandla depression eller inte.

The antitumoural cyanoguanidine CHS 828 has shown promising activity in a number of preclinical and clinical studies. However, the mechanisms underlying the cell death induced by CHS 828 has not been clarified. This thesis describes in vitro studies of the cellular pharmacology of CHS 828.

CHS 828 induced cell death with necrosis like features in the lymphoma cell line U-937 GTB. Addition of 3-aminobenzamide, an inhibitor of ADP-ribosylation, resulted in a decreased sensitivity to CHS 828 and a shift in the mode of cell death towards apoptosis.

Mouse fibroblasts lacking the enzyme PARP-1 were more sensitive to CHS 828 compared to normal fibroblasts. CHS 828 was able to induce p53 in normal fibroblasts but this effect does not seem to be necessary to induce cell death.

Characterization of two CHS 828 resistant cell lines indicated that they were selectively resistant to cyanoguanidines. Known mechanisms of anticancer drug resistance did not seem to account for the cyanoguanidine resistance. One possible resistance mediating protein, which was upregulated in the resistant cells, was epidermal fatty acid binding protein.

A novel high content screening assay was also developed. The assay was shown to be suitable both for screening of potential novel antitumoural substances as well for mechanistic studies. In the assay, CHS 828 induced caspase-3 activity and reduction in mitochondrial membrane potential, both signs of apoptosis, in U-937 GTB cells. However, nuclei in exposed cells did not show nuclear fragmentation, one of the hallmarks of apoptosis.

CHS 828 was also shown to indirectly inhibit the proteasome activity in U-937 GTB cells.

In conclusion, the results presented provide new insights into the metabolic and molecular events involved in cell death induced by CHS 828.

Hypertension is defined as a systolic blood pressure of ≥140 mmHg and/or a diastolic blood pressure of  ≥90 mmHg. The incidence in the Swedish population is 27%. Hypertension becomes one of the leading causes of morbidity and mortality as it increases the risk of developing cardiovascular disease. The cause of hypertension isn’t known yet, but risk factors such as age and obesity have been identified. Hypertension is treated by lifestyle change or by pharmacological treatment. Angiotensen converting enzyme (ACE) inhibitors, angiotensin II receptor blockers (ARBs), calcium channel blockers and thiazide diuretics are the different drugs that are primarily used for treating of hypertension. The most common definition of polypharmacy is  the use of five or more drugs within the same period of time and it’ss associated with an increased risk of interactions and side effects.  Patients with hypertension are characterized by old age, polypharmacy and increased number of hospital stays, making them particularly vulnerable to drug interactions. The blood pressure can be affected by drug-drug interactions between antihypertensive drugs and other medications may increase or decrease the lowering effect of the blood pressure.  The aim of this literature study is to evaluate the common interactions that may occur during treatment with antihypertensive drugs, and the prevalence of these interactions. Five studies were retrieved from the PubMed database and were then analyzed.  The prevalence of drug-drug interactions in each study was 48%, 71.5%, 21.14%, 90.6% and 83.42%. The majority of patients were between 40 and 60 years old and the number of prescriptions per patient was around 5. The results of this literature study show that the prevalence of potential drug interactions is high and that interactions involving atenolol, metoprolol, amlodipine, NSAIDs and insulin are commonly occurring. The results also show that polypharmacy, age and comorbidity significantly increases the risk of drug-drug interactions.
Hypertoni definieras som ett systoliskt tryck som är ≥140 mmHg och/eller ett diastoliskt tryck som är ≥90 mmHg. Förekomsten i Sverige är cirka 27 %. Hypertoni utgör en av de främsta orsakerna till sjuklighet och dödlighet då det innebär en kraftig förhöjd risk att drabbas av kardiovaskulära sjukdomar. Orsaken till hypertoni är ännu inte känd, men riskfaktorer som hög ålder och övervikt har identifierats. Behandling av hypertoni sker genom livsstilsförändringar eller farmakologisk behandling. Angiotensin converting enzyme (ACE)-hämmare, angiotensin-receptorblockerare (ARB), kalciumantagonister och tiaziddiuretika används i första hand. Den vanligaste defintionen av polyfarmaci är  att en patient behandlas med fem eller fler läkemedel samtidigt och detta innebär en ökad risk för interaktioner och biverkningar. Patienter med hypertoni kännetecknas av hög ålder, polyfarmaci och ökad sjukhusvistelse, vilket gör att de är särskilt utsatta för läkemedelsinteraktioner. Kontrollering av blodtrycket kan påverkas genom läkemedelsinteraktioner mellan antihypertensiva läkemedel och andra läkemedel, men också vid samtidig behandling med läkemedel som höjer blodtrycket. Interaktioner mellan antihypertensiva läkemedel och andra läkemedel kan innebära en ökning eller minskning av den blodtryckssänkande effekten. Syftet med denna litteraturstudie är att undersöka vilka vanliga interaktioner som kan förekomma vid behandling med antihypertensiva läkemedel, samt vad prevalensen är för dessa interaktioner. Sju studier hämtades från databasen PubMed och analyserades. Prevalensen av läkemedelsinteraktioner i respektive studie var 48%, 71,5%, 21,14%, 90,6%, 83,42% 55% till 84%, samt 74%.  Majoriteten av patienterna var mellan 40 och 60 år och antalet förskrivna läkemedel per patient var runt 5. Resultatet från den här litteraturstudien visar att prevalensen att potentiellt drabbas av en läkemedelsinteraktion är hög och att interaktioner involverande atenolol, metoprolol, amlodipin, NSAID och insulin är vanligt förekommande. Resultat visar också att polyfarmaci, hög ålder och komborditet signifikant ökar risken att drabbas av läkemedelsinteraktioner.

Bakgrund: Myom är den vanligaste gynekologiska benigna tumören hos kvinnor i fertil ålder. Det finns idag läkemedelsbehandlingar, invasiva ingrepp och icke-invasiva ingrepp vid behandling av myom. Utvecklingen av läkemedel mot myom är lågprioriterat, eftersom de främst är benigna och snarare leder till sjuklighet än dödlighet. De läkemedel som idag används vid behandling av myom är ulipristal, GnRH-agonister, NSAID, tranexamsyra, p-piller eller hormonspiral. Ulipristal och GnRH-agonister har utöver förbättring av symtom som alla de senast nämnda, en effekt på reducering av myomstorlek. Icke-invasiva ingrepp innefattar idag myolys, embolisering av arteria uterina och fokuserad ultraljudskirurgi under MRI-vägledning, framtagna för att slippa operativa ingrepp. Myomektomi och hysterektomi är två operativa ingrepp som genomförs om inga andra behandlingar fungerar, eftersom de medför en längre återhämtningstid och en större risk för komplikationer. Mifepriston är en antiprogesteron, vilket innebär att den hämmar progesterons effekt. Progesteron i sin tur spelar en stor roll i utvecklingen av ett myom. Behandling med läkemedlet har i många olika studier visat goda resultat både i avseende på myomstorlek, symtom, livskvalité och biverkningar. Syfte: Syftet med examensarbetet var att undersöka om mifepriston är ett säkert och effektivt läkemedel vid behandling av myom hos kvinnor. Metod: Examensarbetet är en litteraturstudie och baseras på 6 olika randomiserade kontrollerade vetenskapliga studier som undersökte mifepristons effekt och säkerhet vid behandling av myom. Studierna hämtades från PubMed. Resultat: Samtliga studier visade en reducering i myomvolym, en förbättring av symtom och milda biverkningar med olika doseringar av mifepriston vid behandling av myom. I 2 av studierna undersöktes livskvalitén vilket ökade hos de personer som behandlades med mifepriston. Slutsats: Det är svårt att dra generella slutsatser vilken dos och behandlingstid som är optimala på grund av de olika behandlingstiderna, doseringarna och studieuppläggen som användes i samtliga studier. Dock har mifepriston en god effekt avseende på reducering i myomstorlek, symtom, biverkningar och livskvalité. Det krävs dock fler studier för att säkerställa dosering och behandlingstid samt fler jämförelser med andra behandlingsalternativ som idag finns för behandling av myom.

Niacin som upptäcktes för 70 år sedan är en substans som än i dag studeras mycket kring. Men fortfarande är man inte överens om niacin verkligen har en signifikant effekt på kardiovaskulära sjukdomar. Verkningsmekanismen för niacin är ännu oklar, men forskning pekar på att hämmad syntes av triglycerider resulterar i en minskad LDL-koncentration. HDL-ökningen tros bero på en hämmad nedbrytning av HDL-partiklar, och därmed blir en större mängd HDL kvar i blodet. Nuvarande lipidbehandling består först och främst av en balanserad kost och ökad fysisk aktivitet. Vid otillräcklig effekt sätts läkemedelsbehandling in, som består av statiner, resiner, fibrater och hämmare av kolesterolupptag i tarmen. Syftet med detta arbete är att studera om niacin, både som monoterapi och i kombination med andra lipidsänkande preparat, har en påverkan på blodlipidvärdena HDL, LDL och triglycerider, hos människor. Genom systematisk litteratursökning har artiklar samlats in för att kunna sammanställa resultaten. Alla tio studierna som inkluderades kom fram till att niacin, antingen i kombination med andra lipidsänkande preparat eller som monoterapi, har en signifikant effekt på lipoproteinerna. Resultaten visade en LDL-sänkning upp till 58,5% (i kombination), den högsta HDL-ökning var 50% (monoterapi) och högsta triglycerid-sänkningen var 51,7% (i kombination). Dock är niacins effekt på LDL och triglycerider likartad eller sämre än dagens lipidsänkande preparat. Niacin har däremot en större effekt när det kommer till HDL-ökningen. Därför bör det göras flera och större studier gällande niacins effekt på HDL. Om studierna visar positiva resultat bör det övervägas för användning hos patienter med låg HDL. Dock orsakar biverkningarna en sämre följsamhet, och eventuella analoger till niacin bör utvecklas, detta för att inte gå miste om en eventuellt potent HDL-ökande behandling. Slutsatsen för detta arbete är att i människa har niacin en signifikant effekt på lipoproteinerna LDL och HDL samt lipiden triglycerider. Behandling med niacin leder till en sänkning av LDL och triglycerider samt en markant ökning av HDL.

Introduktion: Läkemedelsbiverkningar delas in i två grupper; typ A och typ B. Typ A-biverkningar är dosberoende medan typ B-biverkningar är idiosynkrasiska och beroende av immunsystemet. Levern är kroppens huvudsakliga metabola organ, och drabbas ofta av läkemedelinducerad toxicitet. Ibland inducerar läkemedelsmetaboliter levertoxicitet, vilket kan medieras av immunsystemet. Karbamazepin är ett antiepileptikum och orsakar levertoxicitet, men den exakta mekanismen är inte klarlagd. Syfte: Syftet med detta arbete är att undersöka om karbamazepins levertoxicitet är beroende av metabolismen av karbamazepin och/eller immunsystemet. Material och metoder: En strukturerad litteraturundersökning utfördes med hjälp av databasen PubMed. 7 artiklar inkluderades i sammanställningen. Resultat: Resultat från in vivo-studier identifierade metaboliter producerade av cytokrom P450-monooxygenaser (CYP450) hos de individer som utvecklade levertoxicitet inducerad av karbamazepin. Samtidigt noterades en ökad nivå CYP3A. Expressionen av en rad immunsystemsmarkörer ökade också vid karbamazepin-inducerad levertoxicitet, exempelvis TNF-α, som leder till apoptos. Slutsats: Utifrån inkluderade studier kan slutsatsen dras att karbamazepins levertoxicitet induceras av dess metaboliter via immunsystemet. Undersökningarna var huvudsakligen associationsstudier, vilket försvårar slutsatser kring kausalitet. Därför behöver ytterligare studier göras så att mekanismen helt kan klargöras.

The purpose of this study was to investigate whether preschool staff who participated in a previous health project retains the good behavior change, and to explore their perceived barriers and opportunities for this. The study was conducted on a "feel good" preschool and the selected population was preschool teachers and/or child minders. The selected method was qualitative, descriptive, and through the interviews the respondents got questions about their behavior changes and their experiences of this. The result of the survey showed that nearly all of the participants in the study had maintained the good behavior changes and most of them had implemented them in their lives. The obstacles that individuals perceived were mainly time constraints and the opportunities they saw were planning, motivation and positive experience. The conclusion was that almost all respondents maintained the previous behavior changes and they have also understood the importance to be physically active and eat a healthy diet. One of the obstacles that individuals considered to be difficult is when there is insufficient time and all of the participants then usually remove the workout. Opportunities that individuals perceived were that planning is important for the success of change unhealthy behaviorsandthat there is a strong motivation. The individuals who participated in the study are driven and well aware of the health benefits they have gained by maintaining their good habits. During the interview it has shown that these people have a very strong will and faith in themselves. They are also motivated to continue their work so that the healthy lifestyle one day will be permanent and a natural part of their lives.

Platelets are important for the healing of damaged blood vessels since they have an importantpart to play in the coagulation process. At the same time, the blood must be kept fluid and notcoagulate at the wrong time. Therefore there are factors that effect the aggregation of plateletsin a positive or a negative way.

Previous investigations have shown that platelets during stirring conditions produce reactiveoxygen species (ROS) that weaken the inhibiting effect of nitric oxides (NO) on platelets andthat the drug Dexamethasone (Dex) can reduce the ROS-production.

The aim of this project was to investigate if glucocorticoids, in this case Dexamethasone,could restore the inhibiting effect of NO on platelets and if there was any decrease in ROS-production.

The result of the ROS-measurements showed a great variance and it was difficult to draw anyconclusions from them, but a clear decrease in ROS, as previous reported, was not shown. In the aggregation experiments the inhibiting effect of NO was observed through the drug S-nitroso-N-acetylpenicillamine (SNAP), a NO-donator.

From the aggregation experiments, the result seemed to be that SNAP during longerincubation time lost its inhibiting effect, probably because the cells become desensitized.With superoxid dismutase (SOD), the effect of SNAP increased, both in the experiment withlonger and shorter incubation times. Dex seemed to reinforce the aggregation in relation toboth SOD and SNAP. To understand this relation further, more investigations must be done.Another interesting experiment would be to do combinations of experiments monitoring bothaggregation and ROS-production at the same time.

Trombocyterna, blodplättarna i blodet, är livsviktiga för att människor inte ska förblöda vid enskada. Samtidigt måste blodet hållas flytande och inte koagulera i onödan och därför finns deti kroppen en mängd faktorer som verkar pro- eller antiaggregerande.

Tidigare undersökningar har visat på att trombocyter har en omrörningsberoendesyreradikalproduktion (ROS) som försvagar kväveoxids (NO) antiaggregerande effekt och attläkemedlet Dexametason (Dex) kan minska denna produktion.

Detta projekt syftade till att ytterligare studera om glukokortikoider, i detta fall Dexametason,kunde återställa NO:s effekt på trombocyterna och om de i någon grad minskaderadikalproduktionen.

Resultatet av ROS-mätningarna blev väldigt varierande och svårtolkade och några säkraslutsatser kunde inte dras, men en tydlig minskning i produktionen som tidigare observeratskunde inte upptäckas. I aggregationsförsöken observerades NO:s inhibitoriska verkan genomS-nitroso-N-acetylpenicillamine (SNAP), en NO-donator.Resultaten tyder på att SNAP under en längre inkuberingstid tappar sin inhiberande förmågapå trombocyterna, vilket förmodligen beror på att cellerna desensibiliseras.Superoxiddismutas (SOD) verkar ha en förstärkande effekt på SNAP oavsett ominkuberingstiden innan dosresponstillsats av trombin är lång eller kort, medan Dex tenderaratt förstärka aggregeringen både i förhållande till SNAP och SOD. För att få mer klarhet omdessa resultat är korrekta måste fler upprepningar göras och dessutom borde man genomförakombinationsförsök där man samtidigt övervakar ROS-produktion och aggregering.

Biological functions within cells and organisms are mainly carried out by the translational products; proteins and peptides. The analysis and characterization of these biomolecules are of great importance for the progress in disease research and biomarker and drug discovery. The term peptidomics was introduced to describe the comprehensive analysis of peptides (e.g. neuropeptides) in biological tissues. In this thesis, a peptidomics approach using nanoflow liquid chromatography coupled to electrospray mass spectrometry (MS) has been developed for detection, identification, and quantification of neuropeptides in different disease models. A thoroughly controlled sample preparation technique and targeted neuropeptide sequence collections have been used to improve sample quality and to increase the number of identified neuropeptides. In particular, neuropeptide changes in experimental models of Parkinson’s disease (PD), with or without L-DOPA treatment, and the effect of antidepressant treatment on neuropeptide expression have been investigated. Several novel, potentially bioactive, neuropeptides have been identified and a number of peptides derived from precursors such as secretogranin-1, preproenkephalin-B, and somatostatin have been found differentially expressed. Some of them represent novel findings, not previously associated with PD or treatment with antidepressants. In addition, MALDI imaging MS (IMS), a technology that permits detection and spatial distribution determination of endogenous compounds and/or administered drugs directly on tissue sections, has been used in both small protein and drug applications. MALDI IMS on tissue samples from experimental models of PD revealed differential expression patterns of two small proteins involved in calcium regulation, PEP-19 and FKBP-12. Biomolecular interaction analysis was performed on FKBP-12 using surface plasmon resonance together with MS and several potential binding partners were identified. In a second approach, MALDI IMS was used to study the distribution of the anticholinergic bronchodilator tiotropium in rat lung following inhalation of the drug. The distribution of the drug was monitored in both MS and MS/MS mode and the levels where linearly quantifiable in the range of 80 fmol – 5 pmol. Conclusively, in this thesis mass spectrometry based technologies have successfully been developed to detect, identify, and characterize small proteins, peptides, and drugs in various tissue samples.

Anti-inflammatory drugs are a widely used class of therapeutic agents, but the use of non-steroidal anti-inflammatory drugs (NSAID) is hampered by their gastrointestinal side-effects. Recent reports that cyclooxygenase-2 inhibitors may cause cardiovascular events underline the importance of identifying new therapeutic strategies for the treatment of inflammation. One such target could be agents modifying the endogenous cannabinoid (endocannabinoid) system, since there is evidence that this system plays a role in our natural defence against inflammation. The levels of the endocannabinoid anandamide (arachidonoyl ethanolamide, AEA) are low under normal conditions, and stand under strict regulatory control of synthesising and degrading enzymes. Fatty acid amide hydrolase (FAAH) is the main enzyme degrading AEA, hydrolysing it to ethanolamine and arachidonic acid. The focus of this thesis lies in exploring the pharmacology of FAAH to evaluate its possibilities as a target for new anti-inflammatory drugs. In Papers I and II, the effects of the ambient pH on the properties of FAAH were investigated, since tissue pH is known to decrease under inflammatory conditions. In homogenates, it was found that the activity of FAAH decreased as the assay pH was decreased, consistent with the known pH profile of the enzyme. More importantly, the sensitivity of the enzyme to inhibition by FAAH inhibitors changed. In particular, the sensitivity of the enzyme to inhibition by the NSAID ibuprofen increased seventeen-fold as the assay pH decreased from 8.37 to 5.28. A similar pattern was found using intact C6 glioma cells when the extracellular, but not the intracellular pH was reduced. Thus, at an extracellular pH value of 6.2, (R)-ibuprofen, (S)-flurbiprofen and (R,S)-flurbiprofen inhibited the metabolism of AEA with IC50 values of 26, 14 and 15 µM, respectively. These values are in theory reachable upon normal dosing of the compounds. In Paper III, the effect of the selective FAAH inhibitor URB597 and the NSAID indomethacin were investigated in vivo upon the oedema response to carrageenan administration in the paw of anaesthetised mice. Both compounds reduced the oedema in a manner completely blocked by the CB2 receptor antagonist SR144528. In Paper IV, the effect of inflammation upon endocannabinoid synthesis was investigated in mice. Lipopolysaccharide-induced pulmonary inflammation was found not to affect the release of AEA to any obvious extent, and did not change the activities of the AEA synthesising enzymes N-acyl transferase or N-acyl phosphatidylethanolamine phospholipase D, or of FAAH in lung tissue. The results of this thesis would suggest that FAAH inhibitors can produce anti-inflammatory effects, and that the endocannabinoid system contributes to the actions of the NSAID indomethacin in the carrageenan model of inflammation, but that an increased endocannabinoid synthesis (a prerequisite for FAAH inhibition as a therapeutic strategy) is not an obligatory response to an inflammatory stimulus.

Background and objectives: Several new psychoactive drugs for the treatment of psychiatric disorders have been introduced onto the market since the late 1980s. Basic aspects of pharmacodynamics and pharmacokinetics (PK) are investigated before approval for general prescription. Thus, a limited number of subjects are exposed to the drug before it is marketed and only sparse measurements of drug concentration are performed during phases II and III of drug development. The objective of this thesis was to provide further descriptive PK and linked patients data in naturalistic clinical settings. The PK of psychoactive drugs was also studied in the elderly and the young, major risk groups that are exposed in normal everyday clinical practice but that are underrepresented in the phases of drug development. The PK-data were to be assessed by samples sent to the Therapeutic Drug Monitoring (TDM) laboratory service. In a subset of individuals, the genotypes of the cytochrome P450 (CYP) enzymes were described. Results: Serum concentration of the parent compound and its metabolites was provided from TDM-data on antidepressant escitalopram (Paper I) and antipsychotic ziprasidone (Paper II). A large interindividual PK variability was found. The daily dose of the drug was higher than the defined daily dose (DDD) for both escitalopram and ziprasidone (median dose 20 mg and 120 mg, respectively). The median number of drugs per patient, apart from the studied drug, was 4 and 3, respectively (range 1-18). If repeated eligible TDM-data were available, change in treatment strategies could be seen between the first and second sample for the patient, and the metabolite/parent compound (M/P) ratio had lower intraindividual than interindividual variation in the escitalopram study but opposite results were found in the ziprasidone study. The prescription of antidepressant drugs (ADs) in the nursing homes studied was 38 % (Paper III). The concentration of the ADs was higher, or much higher, than could be expected from the dose administered in 73 %. The majority of the elderly people were treated with citalopram. No clear time schedule for how long the drug treatment should continue was found in the patients’ current medical record. The median number of drugs per patient apart from the AD was 11 (range 4-19), no monotherapy was found in these patients. The genetically impaired metabolic activity of CYP enzymes correlated to higher drug concentration as expected, in patients medicated with an AD that is substrate for the CYP enzyme genotype. The concentrations of ADs were as expected from the dose administered in 63 % of the children/adolescents evaluated (Paper IV). The majority of TDM samples requested sertraline. PK outcome of sertraline was similar to the results in adult populations. Monotherapy was documented in 49 % (median number of drugs apart from AD was 1 per patient, range 1-7). Changes in treatment strategies were also shown, if repeated TDM-samples were available. The median variation of the M/P ratio for sertraline between the first and the last samples within the same patient was 20 % (the interindividual variation was 37 %). The poor metabolizers (PM) for CYP2D6 medicated with a CYP2D6 substrate had a lower dose than did non-PM for the same drug. Conclusion: These studies provide reference data for the evaluation of the therapeutic response, i.e. a reference range of what is to be expected in a normal clinical setting, as well as the toxicological information concerning the psychoactive drugs studied. When available, the M/P ratio between two patients’ samples may assess patient compliance, as well as drug-drug interactions. Thus, the use of TDM can be beneficial for individual dose optimisation and drug safety, above all in the studied populations, elderly people and children/adolescents, when the selection of doses requires a consideration of PK parameters. TDM may be a tool for research, increasing knowledge of the psychoactive drug in TDM service, as well as toxicology. A more frequent clinical use of TDM and pharmacogenetic testing in clinical practice would contribute to a better quality when treating with psychoactive drugs.

Anandamide (arachidonoyl ethanolamide, AEA) and 2-arachidonoyl glycerol (2-AG) exert most of their actions by binding to cannabinoid receptors. The effects of the endocannabinoids are short-lived due to rapid cellular accumulation and metabolism, for AEA, primarily by the enzymes fatty acid amide hydrolase (FAAH). This has led to the hypothesis that by inhibition of the cellular processing of AEA, beneficial effects in conditions such as pain and inflammation can be enhanced. The overall aim of the present thesis has been to examine the mechanisms involved in the cellular processing of AEA and how they can be influenced pharmacologically by both synthetic natural compounds. Liposomes, artificial membranes, were used in paper I to study the membrane retention of AEA. The AEA retention mimicked the early properties of AEA accumulation, such as temperature-dependency and saturability. In paper II, FAAH was blocked by a selective inhibitor, URB597, and reduced the accumulation of AEA into RBL2H3 basophilic leukaemia cells by approximately half. Treating intact cells with the tyrosine kinase inhibitor genistein, an isoflavone found in soy plants and known to disrupt caveolae-related endocytosis, reduced the AEA accumulation by half, but in combination with URB597 no further decrease was seen. Further on, the effects of genistein upon uptake were secondary to inhibition of FAAH. The ability to inhibit the accumulation and metabolism of AEA was shared by several flavonoids (shown in paper III). In paper IV, the isoflavone biochanin A and URB597 had effects in vivo, in a model of persistent pain, effects decreased by the cannabinoid receptor 1 antagonist AM251. In paper VI, the cellular processing of the endocannabinoid metabolites following degradation was examined, a mechanism poorly understood. It was found that nitric oxide (NO) donors significantly increased the retention of tritium in cell membranes following incubation with either tritiated AEA or 2-AG. Further experiments revealed that the effect of NO donors mainly involves the arachidonate part of the molecules. Inhibition of FAAH completely reduced the effect of NO donors in cells with a large FAAH component, indicating that the effects were downstream of the enzyme. These results suggest that the cellular processing of endocannabinoids can be affected in a manner of different ways by pharmacological manipulation in vitro and that naturally occurring flavonoid compounds can interact with the endocannabinoid system.