No description available.

No description available.

The endogenous opioid system (EOS) including dynorphin opioid peptides and κ-opioid receptor (KOR) plays a critical role in alcohol dependence. Aims of the thesis were to evaluate whether the EOS undergoes molecular adaptations in brain areas involved in cognitive control of addiction in human alcohol dependent subjects, and to analyze the impact of genetic and epigenetic factors on these adaptive changes. The main findings were that (1) the dynorphin/KOR system including PDYN mRNA and dynorphins in the dorsolateral prefrontal cortex (dl-PFC), dynorphins in the hippocampus, and KOR mRNA in the orbitofrontal cortex (OFC), is upregulated in human alcoholics. No other significant changes in the EOS were found. (2) Three PDYN single nucleotide polymorphisms (SNPs), which show the most significant association with alcohol dependence, form CpG sites that are methylated in human brain at different levels. Methylation of the C, non-risk variant of the 3’-untranslated region (3’-UTR) SNP (rs2235749; C>T) was increased in dl-PFC and positively correlated with dynorphins. The DNA-binding factor that differentially targeted the T, risk allele and methylated and unmethylated C allele of this SNP was identified in human brain. We hypothesize that influences of the genetic, epigenetic and environmental factors may be integrated through alterations in methylation of the PDYN 3’jUTR CpG/SNP and, as a consequence, affect PDYN transcription and vulnerability to develop alcohol dependence. (3) The principal component analysis suggested that PDYN expression in the dl-PFC may be related to alcoholism, while in the hippocampus may depend on the genotype of the PDYN promoter SNP (rs1997794; T>C). The T, low risk allele of this SNP resides within non-canonical AP-1-binding element and may be targeted by JUND and FOSB proteins, the dominant AP-1 constituents in the human brain. The T to C transition abrogated AP-1 binding. The impact of genetic variations on PDYN transcription may be relevant for diverse adaptive responses of this gene to alcohol. (4) It was proposed that PDYN transcription may be regulated by intragenic DNA regulatory elements controlling the DNA-protein interactions through formation of non-canonical DNA secondary structures. The dynorphin-encoding sequence in PDYN was found to have potential to form such DNA structure in vitro, and this formation was affected by CpG methylation in this region. This methylation sensitive non-canonical DNA structure formation may be involved in regulation of initiation of PDYN transcription from alternative start sites located within this region, or in splicing of non-canonical mRNA. In conclusion, the dynorphin/KOR system has been identified as the site of robust adaptive changes associated with alcohol dependence in the areas of human brain involved in cognitive control of addiction. Regulation of PDYN was found to be brain area specific, apparently affected by the genetic and epigenetic factors, and possibly dependent on the internal properties of the gene such as its ability to form non-canonical DNA secondary structures.

No description available.

Depriving hormone-dependent breast cancer cells of estrogens has been shown to be a beneficial strategy in the treatment of hormone-dependent breast cancer. The enzymes involved in the biosynthesis of estrogens include: 17[beta]-hydroxysteroid dehydrogenase (17[beta]-HSD), aromatase (AR) and estrone sulfatase (ES). The latter two have been the target of intensive research within our group. Within the current project, we report the synthesis of various types of compounds as potential inhibitors of AR in an effort to determine detailed information regarding the active site of this enzyme; we have also attempted to combine our knowledge of AR and ES in the development of compounds which are able to ihhibit both enzymes. In the first series of compounds to be investigated, compounds were synthesised based on enantiomerically pure forms of the "Evans chiral auxiliary", which is commercially available in both the R and S forms. The oxazolidinone and the phenyl ring systems were derivatised so as to produce N-alkylated compounds which contained a phenylamine moiety for ligating to the Fe of the cytochrome P-450 haem. The compounds were evaluated using a standard literature assay and were, in general, found to be equipotent or more potent than the standard compound aminoglutethimide (AG), with the most potent inhibitor being the pentyi derivative of the R-form (IC50=0.B3±O.05¡..tM). An interesting and different trend was observed 'within the inhibitors based on the S-enantiomer, which were found to be weak inhibitors of AR in comparison to AG - the most potent compound (55) was found to possess an IC50 value of 16.82±0.20¡..tM. The inhibitory activities of both the R- and S-forms were rationalised using the novel substrate-haem complex (SHC) approach. 'From the rationalisation of the inhibitory activity of the oxazolidinone based compounds, the importance of the C(17) position of the steroid backbone was highlighted. As such, we undertook the synthesis and biochemical evaluation of a series of esters based on testosterone. On undertaking the biochemical evaluation of these compounds it was discovered that testosterone possessed potent inhibitory activity (IC50=20.6±O.15¡..tM), and that of the esters, testosterone acetate was shown to have an IC50 value of 4B.7±O.07¡..tM, whereas testosterone decanoate possessed an IC50 value of 164.1±O.5¡..tM. However, the derivatisatian of the alkyl to an aryl carboxylate moiety resulted in one compound which was found to possess highly potent inhibitory activity. That is, testosterone 4-nitrobenzoate was found to possess an IC50 value of B.5±0.05¡..tM. In an effort to synthesise dual inhibitors of AR and ES, we undertook the synthesis of compounds based on the results of our previous studies into ES and the inhibitory activities observed within the testosterone and oxazolidinone based compounds. 'In particular, an attempt was made to incorporate the structural aspects of anastrozole (an aromatase inhibitor) together with a sulfonated phenyl moiety. We synthesised a series of azole based compounds and as expected, these compounds were found to be potent inhibitors of AR, the most potent [1-(4-chloro-benzyl)-1 H-imidazole (BO)] was found to possess an IC50 value of 70±0.5nM. However, the synthesis of the sulfonated compounds proved troublesome and as such no inhibitory data was obtained for these compounds.

In the treatment of hormone-dependent breast cancer, extensive research has been undertaken to produce compounds, which are both potent and selective inhibitors of the cytochrome P-450 enzyme aromatase. However, the use of aromatase inhibitors does not result in the inhibition of all of the biosynthetic processes, which lead to estrogen formation. That is, the enzyme estrone sulfatase converts the stored (sulfated) form of the estrogens to the active (non-sulfated) forms, thereby allowing the stimulation of tumours via a non-aromatase pathway and which, in general, is not blocked by aromatase inhibitors. In an effort to investigate further the structural requirements for estrone sulfatase (and thereby determine the probable mechanism of this enzyme), we designed a range of sulfonated compounds. Here, we report the synthesis and biochemical evaluation of the designed compounds. In general, the results of the study show that the sulfamated compounds are potent inhibitors, possessing greater inhibitory activity than COUMATE, but are weaker than EMATE and the recently reported derivatives of COUMATE, namely 667-COUMATE. However, a small range of compounds (124, 127, 130 and 133) based upon benzoic acid were found to be more potent than the latter compound, in particular, cyclooctyl 4-aminosulfonyl benzoate (133) (IC[sub]50 = 0.17[mu]M) was found to be 1.4 and 2.9 times more potent than 667- COUMATE (IC[sub]50 = 0.23[mu]M) and EMATE (IC[sub]50 = 0.5[mu]M) respectively. The non-sulfamated (for example, 4-nitrophenyl sulfonate based) compounds (52, 58, 61, 103, 131, 146, 170, 173 and 177) were either found to be non-inhibitors or possessed poor inhibitory activity against estrone sulfatase. Consideration of the structure activity relationship of the potent compounds within this study suggests a correlation between two major physicochemical factors [namely, hydrophobicity (logP) and the acid dissociation constant (pK[sub]a) of the parent phenol compound] and the biological activity possessed by sulfamated compounds. However, the present study also suggests another factor, which is presumed to play an important role in stabilising the enzyme-inhibitor complex, namely steric hindrance. That is, consideration of the cyclic ester based compounds against the corresponding straight chain compounds shows that the cyclic ester containing compounds possessed a much greater inhibitory activity. As such, these factors may be combined in an effort to design more potent inhibitors of estrone sulfatase.

Many patients with psychotic symptoms respond poorly to treatment. Various approaches have been made to classify these patients according to treatment response. However, existing classifications have been criticized for various reasons and a new classification system is needed. Further, no satisfactory explanation of the poor treatment response has been apparent. The general aim of this thesis was therefore to develop and validate a new classification method of functional remission in a naturalistic population of patients with psychosis and to utilize this classification to investigate the population from genetic, drug treatment, insight and social network points of view. Data for this cross-sectional study of patients (n=123) attending the Psychosis Outpatient Care clinic in the county of Jönköping, Sweden, were obtained from patient interviews, blood samples and information from patient files. The new classification method CANSEPT, which combines the CAN rating scale (CAN), the UKU side effect rating scale (SE) and the patient’s previous treatment history (PT), showed validity in discriminating the patients and was accepted well by the patients. CANSEPT was used to group the patients in the other studies in this thesis. The results indicated that the gene polymorphism ABCB1 3435T, was related to worse significant social and clinical needs for patients on olanzapine, while the polymorphism DRD2 Taq1 A1 was related to a greater risk of significant side effects; especially if male, or taking strong dopamine D2-receptor antagonistic drugs. Drug treatment factors were also related to treatment response; longer duration of untreated prodromal and early psychosis was seen for patients with current significant social and clinical needs and non-adherence to treatment was associated with worse significant side effects. Worse treatment outcomes also appeared to be associated with smaller social network groups, worse insight into illness, poorer knowledge of warning signs and worse coping strategies. In summary, CANSEPT was shown to be a useful valid, multidimensional tool for classification of treatment response. Gene polymorphisms, duration of untreated illness, non-adherence to treatment, social networks and knowledge should be taken into consideration when investigating inadequate treatment response.

Thesis (Ph. D.)--University of Toronto, 2008. / Includes bibliographical references.

Thesis (M.S.)--University of Wisconsin--Madison, 1965. / eContent provider-neutral record in process. Description based on print version record. Bibliography: l. 186-191.

Thesis (M.S.)--University of Wisconsin--Madison, 1967. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.