Polyunsaturated fatty acids can be bioactivated by two families of dioxygenases, which either contain non-heme iron (lipoxygenases) or heme (cyclooxygenases, linoleate diol synthases and α-dioxygenases). Lipoxygenases and their products play important roles in the pathophysiology of plants and fungi. The only known lipoxygenase with catalytic manganese (Mn-lipoxygenase) is secreted by a devastating root pathogen of wheat, the Take-all fungus Gaeumannomyces graminis. Its mycelia also contains linoleate diol synthase (LDS), which can oxidize linoleic acid to sporulation hormones. Mn-lipoxygenase belongs to the lipoxygenase gene family. Recombinant Mn-lipoxygenase was successfully expressed in the yeast Pichia pastoris with an expression level of 30 mg/L in fermentor culture. The tentative metal ligands of Mn-lipoxygenase were studied by site-directed mutagenesis. The results show that four residues His-274, His-278, His-462 and the C-terminal Val-602 likely coordinate manganese, as predicted by sequence alignments with Fe lipoxygenases. Mn-lipoxygenase (~100 kDa) contains an Asp-Pro peptide bond in the N-terminal region, which appears to hydrolyze during storage and in the acidic media during Pichia expression to an active enzyme of smaller size, mini-Mn-lipoxygenase (~70 kDa). The active form of Mn-lipoxygenase can oxygenate fatty acids of variable chain length, suggesting that the fatty acids enter the catalytic site with the ω-end (“tail first”). Mn-lipoxygenase is an R-lipoxygenase with a conserved Gly316 residue known as a determinant of stereospecificity in other R/S lipoxygenases. The Gly316Ala mutant showed an increased hydroperoxide isomerase activity and transformed 18:3n-3 and 17:3n-3 to epoxyalcohols. The genome of the rice blast fungus, Magnaporthe grisea, contains putative genes of lipoxygenases and LDS. Mycelia of M. grisea were found to express LDS activity. This enzyme was cloned and sequenced and showed 65% amino acid identity with LDS from G.graminis. Take-all and the rice blast fungi represent a constant threat to staple foods worldwide. Mn-lipoxygenase and LDS might provide new means to combat these pathogens.

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.

<p>Supratherapeutic doses of anabolic androgenic steroids (AASs) are administered, not only as performance-enhancing drugs in the world of sports, but also in order to modify behaviour. AAS abusers are at risk of developing serious physical and psychological side effects such as dependence and aggressive behaviour. The aim of this thesis was to investigate the impact of supratherapeutic doses of nandrolone decanoate after subchronic administration on dopamine and serotonin pathways involved in drug dependence and aggression, in the male rat brain.</p><p>Adult male Sprague-Dawley rats received intramuscular injections of nandrolone decanoate (3 or 15 mg/kg) or vehicle once daily for 14 days. Nandrolone decanoate pre-exposure abolished the effect of amphetamine on the 3,4-dihydroxyphenylacetic acid (DOPAC) tissue level in the hypothalamus and on the DOPAC/dopamine ratio in the hypothalamus and the hippocampus. A significant decrease of the basal extracellular DOPAC and homovanillic acid (HVA) levels could be detected in the nucleus accumbens, which remained low during the first hour following the amphetamine challenge. Nandrolone decanoate significantly reduced the activity of both monoamine oxidase A and B (MAO-A and -B) in the caudate putamen and amygdala. The gene transcript levels of MAO-B, and the dopamine D1 and D4 receptors were altered in limbic regions. No changes in transcriptional levels could be detected among the serotonin receptor genes examined. However, the density of the serotonin transporter protein was elevated in a range of aggression-related brain regions.</p><p>Taken together, subchronic administration of nandrolone decanoate causes dopaminergic and serotonergic dysregulations in distinct brain regions. These areas of the brain are involved in the development of drug dependence and expression of impulsive and aggressive behaviours. These results may contribute to explain some of the behavioural changes often reported in AAS abusers, such as polydrug use and impaired impulse control.</p>

The persistent and toxic perfluoroalkyl acids (PFAAs) are ubiquitously present in the environment and reach humans predominantly via food and drinking water. The aim of the present study was to investigate the effect of low-grade (&lt;10 ng L-1 of single PFAAs) contaminated PFAAs drinking water on serum PFAA concentrations in a representative Swedish adolescent population, and to examine the influence of potential determinants on the variation of the PFAAs serum concentrations. This was done by using multivariate regression analysis on the possible determinants of blood serum PFAA concentrations in 479 Swedish adolescents, 10 to 21 years of age, who had left complete dietary and life style information in 2016-17 in the nation-wide food consumption survey Riksmaten Ungdom. Raw and drinking water samples (DW) from water treatment plants (WTPs) that delivered DW to participants schools were sampled in 2018, analysed for PFAAs, and used for assessing the participants DW PFAA exposure. Maternal education level and maternal birth country, consumption of fish, as well as age and sex were significantly associated with the participants PFAAs serum concentrations. DW concentrations as low as &lt;1 ng L for PFOA and PFHxS, &lt;0.45 ng L-1 for PFNA and &lt;4 ng L-1 for PFOS were significantly associated with increased adolescent serum concentrations of the PFAAs in question, which suggests that low-grade contaminated drinking water is an important exposure route for Swedish adolescents. For risk assessment purposes, it was investigated whether parts of the adolescent population exceeded the serum PFOS and PFOA concentrations corresponding to the current health-based reference intakes as assessed by the European Food Safety Authority (EFSA) and the U.S. Agency for Toxic Substances and Disease Registry (ATSDR). Around 1.7% and 2.7% of participants had PFOS serum concentrations exceeding serum levels used to derive the tolerable daily intake (TDI) (EFSA) and the minimum risk level (MRL) (ATSDR), respectively and a cause for concern was consequently identified. However, the high serum concentrations of participants exceeding the TDI and MRL serum concentrations belong to participants suspected to have been previously exposed to highly contaminated drinking water and not from consuming foods and beverages containing background concentrations of PFAAs.

Introduktion: Bensodiazepiner har ända sedan de introducerats till marknaden varit mycket populära och är bland de mest förskrivna läkemedlen. Trots populariteten samt den höga användningen är bensodiazepiner en problematisk läkemedelsgrupp med anledning till deras höga beroendepotential. Bensodiazepiner ska enligt behandlingsrekommendationer endast förskrivas som sista alternativ och användas under en begränsad period för att undvika beroendeutveckling, vilket oftast inte blir fallet i primärvården. Syfte: Syftet var att undersöka hur användningen av bensodiazepiner har sett ut mellan 2006 och 2014 och om beroendepotentialen var en möjlig faktor till hypotesen att användningen minskat– för att uppmärksamma problem med användningen av dessa läkemedel. Material och metoder: Artiklar hämtades från databaser som PubMed samt Google Scholar. Statistiken över användningen hämtades från Socialstyrelsens statistikdatabaser och presenterades i form av tabeller och grafer. Resultat: Denna studie undersökte om just beroendepotentialen var en möjlig faktor till hypotesen att användningen minskat – och hypotesen stämmer. Statistiken visade att användningen av många bensodiazepiner har minskat markant under den perioden som undersöktes; som mest troligt beror på de nya riktlinjerna och behandlingsrekommendationerna som tagits fram mellan 2009-2010. Trots att användningen av bensodiazepiner minskat stadigt har istället en ny trend upptäckts; att bensodiazepinbesläktade substanser, som zopiklon, ökar markant. Detta innebär att behoven av farmakologisk behandling fortfarande är densamma då användningen av bensodiazepiner och bensodiazepinbesläktade läkemedel inte har varierat signifikant under den undersökta perioden. Konklusion: Nya selektiva bensodiazepiner med obefintlig risk för beroendeutveckling och smalare biverkningsprofil är under utveckling; dessa kommer med störst sannolikhet lösa många av beroendeutvecklings-problemen som för närvarande uppstår vid användning av bensodiazepiner och besläktade substanser i primärvården. / Introduction: Ever since benzodiazepines were introduced to the market they have been very popular and are among the most prescribed drugs. Despite the popularity and the high usage of benzodiazepines, this group of drugs is problematic due to their high dependence potential. Benzodiazepines should according to treatment guidelines only be prescribed as a last option and be used for a limited period of time to avoid the development of dependency - which is often not the case in primary care. Aim: The aim was to examine the usage of benzodiazepines between 2006 and 2014 and if the dependency potential was a possible factor to the hypothesis that the use has decreased – in order to draw attention to the problems associated with benzodiazepines. Methods: The articles were retrieved from the databases PubMed and Google Scholar. Statistics on the usage were taken from the Board's statistical databases. Results: This study investigated if the dependency potential was a possible factor to the hypothesis that the usage of benzodiazepines have decreased – and the hypothesis is correct. Statistics showed that the usage of many benzodiazepines have declined significantly during the period examined; most likely due to the new guidelines and treatment recommendations developed between 2009-2010. Although the usage of benzodiazepines declined steadily; a new trend was discovered, namely the prominent increased usage of benzodiazepine-like drugs such as zopiclone. This means that the needs of pharmacological treatment is unchanged since the usage of benzodiazepines and benzodiazepine-like drugs have not varied significantly during the examined period. Conclusion: New selective benzodiazepines with non-existent risks of development of dependency as well as a narrower side effect profile is under development. These will most likely resolve many of the dependency issues currently arising from the usage of benzodiazepines and related substances in primary care.

Familial hypercholesterolemia is a genetic disease affecting about 10 million people around the world. Those who carry the disease have a very high risk of developing cardiovascular diseases and commonly encounter myocardial infarction at the early age of 40. Therefore, a diagnosis and immediate treatment are very important for these patients. Despite many combinations of available drugs, there are many patients who still cannot reach the desired cholesterol levels. Mipomersen is a new lipid-lowering drug which inhibits the synthesis of apolipoprotein B, a common component of lipoproteins such as low-density lipoprotein. Inhibition of this protein leads to reduced production of these lipoproteins and reduces the risk of cardiovascular diseases. The drug is currently only indicated for treating patients with homozygous familial hypercholesterolemia.  Unfortunately, there have been many reports of adverse events in patients using mipomersen which has proven problematic.         The aim of this thesis is to analyze the efficacy and safety of mipomersen when treating patients with familial hypercholesterolemia. This has been done by searching for five clinical trials in the database Web of Science. The studies were required to include patients with familial hypercholesterolemia, use mipomersen as the study drug and analyze its effect and safety.   The studies showed that mipomersen has a very good effect in decreasing low-density lipoproteins as well as other lipoproteins in comparison to placebo. Many of the patients who were treated with mipomersen displayed several adverse events and the most common were injection-site reaction and influenza-like symptoms. Elevated levels of aminotransaminase and increased fat deposit in the liver were also common. Based on the five clinical trials analyzed in this thesis, mipomersen is an effective lipid-lowering drug which reduces low density lipoprotein cholesterol, apolipoprotein B and lipoprotein (a) in patients with familial hypercholesterolemia. Elevations in alanine aminotransferase and aspartate aminotransferase are common in patients treated with mipomersen. This could indicate a negative impact on the liver. To be more certain of its safety profile, more research could be needed. There are however, new treatments that combines statins and a proprotein convertase subtilisin/kexin 9 inhibitor, which could be the future of lipid-lowering treatments and mipomersen would then likely be substituted.

<p>Mantle cell lymphoma (MCL) is a non-Hodgkin B-cell lymphoma with very bad prognosis. The genetic hallmark of MCL, is the translocation t(11;14)(q13;q32) which leads to overexpression of cyclin D1, a D-type cyclin that is not usually expressed at high levels in normal B lymphocytes.</p><p> </p><p>Previous studies indicate that cannabinoid receptors are expressed in lymphoma and have shown that lymphoma cell death is induced as a result of exposure to cannabinoids (ligands).</p><p> </p><p>The aim of this diploma work was to combined cytostatics with the cannabinoid receptor ligand R (+)-Methanandmide (R-MA). Our data suggest that combination treatment with cytostatics and R-MA induces synergistic effects in most cases.</p>

<p>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.</p><p>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.</p><p>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.</p><p>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.</p><p>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.</p> / <p>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.</p><p>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.</p><p>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.</p><p>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.</p>

<p>Eukaryotic gene transcription plays a regulatory role in mammalian developmental processes. It has been shown that transcriptional control is an important mechanism for specification of neurotransmitter phenotypes. In the mammalian central nervous system, the transcription factor AP-2 family is one of the critical regulatory factors for neural gene expression and neuronal development. It has been shown that several genes in the monoaminergic systems have AP-2 binding sites in regulatory regions, suggesting a regulatory role of AP-2 also in the adult brain. Brainstem monoamines are implicated in the expression of personality traits and imbalances in these systems may give rise to psychiatric disorders. </p><p>The gene encoding AP-2β includes a polymorphic region consisting of a tetranucleotide repeat of [CAAA]_4-5 in intron 2. Studies on AP-2β genotype in relation to personality and platelet MAO activity, a trait-dependant marker for personality, are presented in this thesis. Furthermore, correlations between brainstem levels of AP-2α and AP-2β and monoamine turnover in projection areas in rat forebrain are reported. These results strengthen the notion that the AP-2 family is important regulators of the monoaminergic systems in the adult brain. Furthermore, two studies are presented in this thesis with analyses indicating a role for AP-2 in the molecular mechanism of antidepressant drugs.</p><p>Altogether, this thesis presents data supporting our notion that the transcription factor AP-2 family is involved in the regulation of the monoaminergic systems both pre- and postnatally, and, therefore, might be involved in the pathophysiology of neuropsychiatric disorders.</p>

<p>Plasma activity of the enzyme semicarbazide-sensitive amine oxidase (SSAO; EC.1.4.3.6) has been reported to be high in disorders such as diabetes mellitus, chronic congestive heart failure and liver cirrhosis. Little is known of how the activity is regulated and, consequently, the cause for these findings is not well understood. Due to the early occurrence of increased enzyme activity in diabetes, in conjunction with the production of highly cytotoxic substances in SSAO-catalysed reactions, it has been speculated that there could be a causal relationship between high SSAO activity and vascular damage. Aminoacetone and methylamine are the best currently known endogenous substrates for human SSAO and the resulting aldehyde-products are methylglyoxal and formaldehyde, respectively. Both of these aldehydes have been shown to be implicated in the formation of advanced glycation end products (AGEs).</p><p>This thesis is based on studies exploring the regulation of SSAO activity and its possible involvement in the development of vascular damage. The results further strengthen the connection between high SSAO activity and the occurrence of vascular damage, since type 2 diabetic patients with retinopathy were found to have higher plasma activities of SSAO and lower urinary concentrations of methylamine than patients with uncomplicated diabetes. From studies on mice, it was also found that an SSAO inhibitor potently reduces the incorporation of methylamine-metabolite in the tissues. By quantifying SSAO-gene expression in alloxan-induced diabetes, increased transcription could be ruled out as a cause for the increased enzyme activity, thereby opening up for the possibility that the activity is regulated post-translationally. In fact, increased enzyme activity in adipose tissue was accompanied by decreased mRNA-levels, suggesting that the gene expression could be negatively controlled by the enzyme activity.</p>