myo-lnositol (mIns), a natural component of the human diet and essential precursor of several signalling pathways, including that of G protein-coupled receptors, has also been shown to be effective in the treatment of psychiatric disorders such as depression, obsessive compulsive disorder and panic disorder. Most likely since mlns is a simple isomer of glucose, no serious side effects have been reported with its use, even at high oral doses of mlns. Previous studies suggest that the therapeutic action of mlns may include reduced serotonin 5HTzA and muscarinic acetylcholine receptor function. An important signal transduction system that may possibly be involved in the mechanism of action of antidepressants is phosphoinositide (PI) turnover. In this signalling system PI-phospholipase C (PLCpl), that is implicated in the in the mechanism of action of antidepressants and anxiolytics, is activated. The mechanism of action of mlns, however, still remains elusive and needs further investigation. In this study a possible modulatory role of 24-hour pre-treatment of human neuroblastoma cell line (SH-SY5Y) with mlns on mRNA levels and protein expression of phospholipase C-p1 (PLCP1) and glycogen synthase kinase 3P (GSK3p) was investigated. The effects of mlns were also compared to that of other prototype antidepressants, such as fluoxetine (a selective serotonin reuptake inhibitor), imipramine (a tricyclic antidepressant), lithium and another drug with potential antidepressant effects, sildenafil (phosphodiesterase 5-type (PDE5) inhibitor). Real-time reverse transcription Polymerase Chain Reaction (RTPCR) was performed in order to investigate the mRNA levels, while protein expression in membranes and the cytosol fraction of cells were quantified with Western blots. The expression of PLCPl was decreased after pre-treatments with imipramine or myoinositol in combination with fluoxetine. In addition, sildenafil alone or in combination with myo-inositol, also decreased the expression of membrane-bound PLCp1. However, a 24- hour pre-treatment with lithium did not alter PLCPl expression significantly. Determined mRNA levels for the expression of PLCPl were consistent in these findings, except for the inhibition of the mRNA for the expression of PLCPl also after lithium treatment. The reduced PLCpl mRNA levels after lithium pre-treatment may suggest the involvement of posttranscriptional modification (or delayed translational effects) of PLCpl after lithium treatment. The data from the current study suggest that antidepressant action may include downregulation of PLCPl expression and that modulators of the nitric oxidecGMP pathway (e.g. sildenafil as a PDE5 inhibitor) may exhibit similar properties. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2005.

Made available in DSpace on 2016-06-02T20:21:34Z (GMT). No. of bitstreams: 1 dissHRS.pdf: 548212 bytes, checksum: 1c4f402bcc4318a0c91d0661cf16825d (MD5) Previous issue date: 2003-02-25 / Hemoglobin electrophoretic patterns, oxygen equilibrium experiments using unfractionated blood, stripped hemoglobin at 4oC, 21oC, 40oC, as well as the effect of inositol hexaphosphate (IHP) and 2,3-biphosphoglycerate (BPG), and the qualitative and quantitative determination of intraerythrocytic organic phosphate were done, in three species of Aves: Athene cunicularia – owl (Order Strigiformes, Family Strigidae), Coragyps atratus - vulture (Order Ciconiformes, Family Cathartidae) and Mimus saturninus - mocking bird (Order Passariformes Family Mimidae). On starch gel electrophoresis, pH 8.6, A. cunicularia and C. atratus showed one slow anodic major component and one fast minor components, a typical bird hemoglobin pattern. On the same electrophoretic conditions, M. saturninus showed two anodic major components, instead one major and one minor. Stripped hemoglobins at the three temperatures, in the presence of IHP, BPG, and blood showed normal Bohr effect and n values (Hill plots) were close to one, indicating very low or no cooperativity. Oxygen affinities were temperature-dependent (as lower the temperature higher is the affinity), BPG modulation was low, IHP was very effective as negative modulator. Vulture hemoglobin showed the highest P50 value, while the owl and the mocking bird presented lower and similar oxygen affinity to each other. No AMP, GTP and BPG were found, only ADP, ATP and inositols were detected in the erythrocytes of the three species. Inositol pentaphosphate (IP5) was the most abundant intraerythrocytic organic phosphate in the 3 species. A. cunicularia showed also inositol tetraphosphate and C. atratus and M. saturninus showed IHP, besides IP5. Although the three species belong to different Orders, have different habit; the functional properties of their hemoglobins are very similar. One hypothesis to explain such similarity is that the avian system to deliver and discharge oxygen to the tissues are so efficient, that works properly in a wide range of situations / Foram analisados, o padrão eletroforético, a finidade das hemoglobinas (Hbs) em experimentos de equilíbrio de oxigenação em sangue, hemoglobinas stripped à 4oC, 21oC e 40oC, o efeito de fosfatos orgânicos inositol hexafosfato (IHP) e 2,3 - bifosfoglicerato (BPG), bem como a determinação qualitativa e quantitativa de fosfatos orgânicos em três espécies de Aves: Athene cunicularia – coruja-buraqueira (Ordem Strigiformes, Família Strigidae), Coragyps atratus - urubu (Ordem Ciconiformes, Família Cathartidae) e Mimus saturninus - sabiá-do-campo (Ordem Passariformes Família Mimidae). O padrão eletroforético em gel de amido (pH 8,6), para A. cunicularia e C. atratus, mostra a presença de um componente maior lento, e outro menor rápido, padrão típico de hemoglobinas de aves. Para M. saturninus o padrão eletroforético mostra dois componentes maiores, um rápido e outro lento. As hemoglobinas stripped, nas três temperaturas, na presença de IHP, 2,3 - BPG e em sangue mostraram efeito Bohr normal e valores de n (Hill) próximos de um, indicando cooperatividade muito baixa ou ausente. A afinidade das Hbs ao oxigênio mostrou - se maior a medida que se diminuiu a temperatura. O 2,3 - BPG praticamente não modulou as Hbs, já o IHP atuou como modulador negativo efetivo das Hbs. Não foi detectado concentrações de AMP, GDP, GTP e 2,3 - BPG. Foram detectados somente ADP, ATP e inositóis nos eritrócitos das três espécies. O inositol pentafosfato (IP5) foi o fosfato intraeritrocitário mais abundante nas três espécies. A. cunicularia mostrou também inositól tetrafosfato (IP4) e C. atratus e M. saturninus, mostraram IHP. Apesar das três espécies estudadas pertencerem a ordens diferentes, com tipos de vôo e hábitos distintos, as propriedades funcionais de suas hemoglobinas são muito semelhantes. Já que as três espécies estudadas, com hábitos tão diferentes, apresentam características funcionais tão semelhantes, é possível que esses sistemas sejam tão eficientes que não necessitem modificações significativas para cumprir seu papel de fornecer oxigênio aos tecidos, numa gama grande de situações ecológicas

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Breast cancer (BC) is one of the most common malignancies in women. Death results from treatment failure and metastatic disease. Thousands of lives might be saved if it was possible to detect and eliminate occult metastatic cells before they become clinically evident. Therefore, there is a critical need to identify new markers to improve treatment options for these patients. Genomic instability is the earliest indication of breast cancer and the use of genomic methodologies is a progress towards early detection and treatment, through the identification of biomarkers that can be translated into novel therapy targets. The interferon regulatory factor-1(IRF-1) gene, localized on chromosome 5q31.1, is believed to act as a tumor suppressor gene in breast cancer. The IRF-1 was found to be inactivated by single nucleotide polymorphism (SNP) in breast cancer suggesting that the loss of its function might be critical to the development of the disease. The phosphatidylinositol 3-kinase (PIK3) signaling pathway mediates key cellular functions and alterations of genes in this pathway, including PIK3CA, serine-threonine protein kinases (AKT1and AKT2), phosphatase and tensin homolog (PTEN), fibroblast growth factor receptor 2 (FGFR2) and ERBB2, whose expression have been demonstrated to be altered in breast cancer patients. In addition, these genes are linked to treatment resistance. vi In this study, we have investigated allelic loss of IRF-1 gene in primary tumors obtained from patients undergoing mastectomy at Groote Schuur hospital (Cape Town, South Africa). These samples were then further analyzed for the DNA copy number changes of specific genes involved in the PIK3/AKT signaling pathway. Statistical analysis has been performed in order to correlate genomic findings with clinical-histopathological and follow up information from the patients and to establish whether these genes can predict prognosis. Our data analysis has indicated that 46 cases (45.5%) out of 101 cases were informative for the IRF-1 dinucleotide marker used for LOH analysis (Figure 3.1). LOH was detected in 23 of these informative cases (23/46; 50%). No statistical significance was found between LOH at the IRF-1 locus and age (≤50 years or >50 years) (P value = 1.0000) and earlier stage (Stages I and II) (P value= 0.4982) based on Fisher’s exact test. Patients presented a high level of DNA copy number changes in genes involved in the PIK3/AKT pathway. The most frequent changes were observed in the PIK3CA and PTEN genes. PIK3CA presented high copy number in 36.8% of the cases. PTEN was observed with low copy number in 47.5% of the cases. This dissertation shows the effectiveness of genomic methodologies as means for the detection of early breast cancer progression in South African women. The PIK 3/AKT genes can validate the usefulness of breast cancer therapies.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2015-11-10T08:33:00Z No. of bitstreams: 1 texto completo.pdf: 2551787 bytes, checksum: 71ddb0374c66aab0bc01094fd2206080 (MD5) / Made available in DSpace on 2015-11-10T08:33:00Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2551787 bytes, checksum: 71ddb0374c66aab0bc01094fd2206080 (MD5) Previous issue date: 2014-02-25 / Fundação de Amparo à Pesquisa do Estado de Minas Gerais / A enzima mio-inositol-1-fosfato sintase (MIPS1) (E.C.5.5.1.4) catalisa a conversão irreversível de D-Glicose-6-P para 1-L-mio-inositol-1-P. O mio-inositol desempenha papel de destaque no metabolismo vegetal, fornecendo inositol e inositídeos em processos metabólicos essenciais à formação do vegetal. Dentre as várias vias nas quais o mio-inositol está envolvido algumas são alvo do melhoramento vegetal, a exemplo a manipulação do conteúdo de ácido fítico e de oligossacarídeos em sementes e açúcares estruturais na parede celular. Até o presente momento, plantas MIPS silenciadas não foram caracterizadas em detalhes, principalmente em suas estruturas vegetativas. Com isso, o presente trabalho objetivou caracterizar tomateiros ‘Moneymaker’ com diferentes níveis de silenciamento de MIPS1. Para a indução do silenciamento utilizou- se tranformação genética via Agrobacterium tumefaciens contendo plasmídeo pCambia com construção tipo intron hairpin de fragmentos do gene MIPS de Glycine max (∆GmMIPS) sob controle do promotor CaMV 35S. Avaliou-se o efeito do silenciamento de MIPS1 em plantas transgênicas matrizes T0 e progênies T1 e T2 quanto ao desenvolvimento de frutos e sementes, conteúdos de ácido fítico, açúcares, pigmentos fotossintéticos, respostas fisiológicas mediante a avaliação das trocas gasosas, fluorescência da clorofila a, atividade de enzimas do estresse oxidativo e caracterização do perfil metabólico e constituintes de parede celular. Os resultados apontaram eficiência da construção transgênica na indução do silenciamento. O silenciamento ocasionou redução do conteúdo de ácido fítico em sementes, redução da quantidade de sementes por fruto e redução no desenvolvimento do fruto; redução na germinação e desenvolvimento de plântulas provindas de sementes normais nas progênies T0 e T1. O silenciamento também ocasionou alterações no conteúdo de açúcares, em folhas e sementes nas progênies T0 e T1 e reduziu a firmeza da epiderme e do pericarpo em frutos da progênie T1. Tal redução na firmeza pode estar relacionada à composição ou estrutura das paredes celulares; porém não foram identificadas reduções no conteúdo de açúcares de parede celular e celulose em pericarpos e folhas de plantas T2, mas sim acumúlo de ácidos urônicos, tanto em pericarpo quanto em folhas. Em relação ao aparato fotossintético, o silenciamento não causou alterações. O conteúdo de pigmentos fotossintéticos, parâmetros de trocas gasosas e fluorescência da clorofila a foram semelhantes aos apresentados pelas plantas não transformadas, Outra evidência que o silenciamento de MIPS não causa distúrbios no corpo vegetativo foi a atividade de enzimas do estresse oxidativo que, em condições ótimas, não apresentou diferenças. As alterações encontradas no corpo vegetativo tiveram estreito envolvimento com o metabolismo de inositol, como conteúdo de açúcares (glicose-6P, glicose e sacarose) em horários de intensa atividade fotossintética. A metabolômica possibilitou uma análise mais integrativa em que ficou evidenciada a redução de alguns aminoácidos, acúmulo de açúcares e redução de açúcares alcoóis, todos estreitamente relacionados ao metabolismo de inositol em folhas. Em frutos, verificou-se influência mais expressiva da MIPS, com alterações em todas as quatro idades avaliadas (10, 15, 50 e 60 dias após a antese), tanto no pericarpo quando nas sementes. O maior número de alterações foi encontrado nas sementes, caracterizadas pela redução do conteúdo de aminoácidos, de intermediários do ciclo dos ácidos tricarboxílicos (TCA), de ácidos orgânicos e de açúcares alcoóis. Os açúcares, por sua vez, apresentaram comportamento variado, dependendo da sua relação com o metabolismo de inositol, onde a frutose e a trealose apresentam acúmulo, enquanto a glicose e a sacarose apresentam padrão mais variado em decorrência do envolvimento em uma rede mais complexa de regulação. Em pericarpos, o perfil metabólico foi respondente as variações de desenvolvimento, porém apresentando alterações no conteúdo de açúcares. Com esse estudo podemos afirmar que a construção tipo intron hairpin de fragmentos do gene MIPS de Glycine max (∆GmMIPS), sob controle do promotor CaMV 35S, ocasiona silenciamento em tomateiros e as plantas silenciadas tiveram o conteúdo de metabólitos influenciados pelo conteúdo de mio-inositol ou expressão da MIPS alterado, tanto na parte vegetativa quanto na reprodutiva. Registra-se, também, que o silenciamento de MIPS não foi capaz de alterar a atividade fotossintética ou causar direta ou indiretamente estresse oxidativo no corpo vegetativo em plantas MIPS silenciadas. / Myo-inositol phosphate synthase (MIPS1) (E.C.5.5.1.4) catalyzes the irreversible conversion of D-Glucose-6-P to 1-L-myo-inositol-1-P. Myo-inositol plays an important role in plant metabolism, providing inositides and inositol for essential metabolic processes in the formation of the vegetative and reproductive plant body. Among the pathways in which the myo-inositol is closely involved, some are target of plant breeding, such as genetic manipulation of phytic acid and oligosaccharides contents in seeds and structural sugars in the cell wall. To date, MIPS silenced plants have not been characterized in detail, especially in their vegetative structures. Thus, the present study aimed to characterize tomato 'Moneymaker' with different levels of silencing MIPS1. For induction of gene silencing was used transformation via Agrobacterium tumefaciens containing the plasmid pCambia with construct intron-hairpin-RNA type whith MIPS gene fragments of Glycine max (ΔGmMIPS) under control of the CaMV 35S. The effect of MIPS1 silencing in primary transgenic plants T0 and T1 and T2 progenies was evaluated for the development of fruit and seeds, phytic acid and sugar contents, photosynthetic pigments, physiological responses by evaluating gas exchange, chlorophyll a fluorescence, oxidative stress enzyme activity, metabolic profile characterization and cell wall constituents. The results showed the efficiency of the transgenic construct to induce silencing. The silencing resulted in reduction of phytic acid content in seeds, reducing the amount of seeds and a reduction in fruit development, reduction in germination and seedling development stemmed from normal seeds in T0 and T1 progeny. The silencing also induced changes in the content of sugars in leaves and seeds in T0 and T1 progeny and reduced firmness of the epidermis and pericarp in fruits of T1 progeny. This decrease in firmness may be related to the composition or structure of cell walls, but no reductions were identified in the content of sugars and cellulose in cell wall pericarp and leaves of T2 plants, but accumulation of uronic acids, both in pericarp as sheets. In relation to the photosynthetic apparatus, silencing caused no changes. The content of photosynthetic pigments, gas exchange parameters and chlorophyll fluorescence were similar to those presented by the non- transformed plants, Further evidence that silencing of MIPS does not cause disturbances in the vegetative body was the activity of enzymes of the oxidative stress that in optimal conditions, showed no differences. However, the changes found in the vegetative body had close involvement with the metabolism of inositol, as content of sugars (glucose- 6P, glucose and sucrose) in times of intense photosynthetic activity. The metabolomics allowed a more integrative analysis in which there is an evident reduction of some amino acids, accumulation of sugars and sugar alcohols reduction, all closely related to the metabolism of inositol sheets. In fruits, there was more significant influence of MIPS, with changes in all four ages evaluated (10, 15, 50 and 60 days after anthesis), both when the seed pericarp. The largest number of changes were found in the seeds, characterized by the reduction of the content of amino acids, intermediates of the citric acid cycle (TCA), organic acids and sugar alcohols. Sugars, in turn, showed varied behavior depending on their relationship with the metabolism of inositol, where fructose and trehalose accumulation feature, while glucose and sucrose have more varied pattern as a result of being involved in a more complex regulatory network. In pericarp, the metabolic profile was respondent variations in development, but with alterations in sugar content. With this study we can see that the construction type intron hairpin fragment of MIPS Glycine max (ΔGmMIPS) gene under control of the CaMV 35S promoter, causes silencing in tomato plants were silenced and content of metabolites influenced the content of myo-inositol or altered expression of MIPS, both in vegetative and in reproductive part. Join also the silencing of MIPS was not able to change the photosynthetic activity or directly or indirectly cause oxidative stress in the vegetative body MIPS-silenced plants.

Chemical substances that are either produced by nature or by humankind have a consequence on the environment when they are accumulated to a great extent. One of nature’s major problem is that of the eutrophication of surface waters, which is caused by inorganic and organic phosphorus compounds and, in particular, from a group known as Inositol Phosphates. There are six different forms of these inositols and are commonly found in soil and sediment samples. The conducted project was aimed to develop an analytical method that could efficiently analyze and separate all six with repeatable results. As such, soil samples were collected two times from two forest locations and two crop field locations. The first time the soil samples were left overnight to dry in a drying oven while on the second time, the samples left to dry at room temperature. When the samples were considered dry enough, they processed to reach grain size and extracted with a mixture of NaOH and Titriplex® III. The extracted soil samples, the standard solutions containing inositols, and the spiked extracted soil samples with inositol solution were all analyzed with an instrumental combination of a Capillary Electrophoresis instrument coupled manually to an Electrospray mass spectrometer, where the first was operated at reversed mode and the second at negative mode. To achieve the best feasible separation, several background electrolyte solutions were created along with a large number of sheath liquids regulated by an LC pump, two Capillary Electrophoresis methods, and twenty-five distinct MS methods, all tested through extensive screening to obtain the best possible combination of parameters. Out of the obtained results from the runs, four background electrolyte solutions, two MS methods, one sheath liquid controlled by one specific flow rate, and one Capillary Electrophoresis method exhibited promising potentials with a satisfying outcome. However, the intense pulsation of the spray cone observed for many of the runs, the manual protrusion of the Capillary Electrophoresis fused silica capillary, and some random errors, the repeatability of the method is called into question.

The release of chemical transmitter from nerve terminals is critically dependent on a transient increase in intracellular Ca2+.6,25 The increase in Ca2+ may be due to influx of Ca2+ from the extracellular fluid15 or release of Ca2+ from intracellular stores such as mitochondria.1,8,18 Whether Ca2+ utilized in transmitter release is liberated from organelles other than mitochondria is uncertain. Smooth endoplasmic reticulum is known to release Ca2+, e.g., on activation by inositol trisphosphate or cyclic adenosine diphosphate-ribose,2 so the possibility exists that Ca2+ from this source may be involved in the events leading to exocytosis. We examined this hypothesis by testing whether inositol trisphosphate and cyclic adenosine diphosphate-ribose modified transmitter release. We used liposomes to deliver these agents into the cytoplasmic compartment and binomial analysis to determine their effects on the quantal components of transmitter release. Administration of inositol trisphosphate (10-4M) caused a rapid, 25% increase in the number of quanta released. This was due to an increase in the number of functional release sites, as the other quantal parameters were unaffected. The effect was reversed with 40min of wash. Virtually identical results were obtained with cyclic adenosine diphosphate-ribose (10-4M). Inositol trisphosphate caused a 10% increase in quantal size, whereas cyclic adenosine diphosphate-ribose had no effect. The results suggest that quantal transmitter release can be increased by Ca2+ released from smooth endoplasmic reticulum upon stimulation by inositol trisphosphate or cyclic adenosine diphosphate-ribose. This may involve priming of synaptic vesicles at the release sites or mobilization of vesicles to the active zone. Inositol trisphosphate may have an additional action to increase the content of transmitter within the vesicles. These findings raise the possibility of a role of endogenous inositol phosphate and smooth endoplasmic reticulum in the regulation of cytoplasmic Ca2+ and transmitter release.

One of the consequences of G-protein-coupled receptor activation is stimulation of phosphoinositol metabolism, leading to the generation of IP 3 and its metabolites 1,3,4,5-tetrakisphosphate (IP4) and inositol 1,2,3,4,5,6-hexakisphosphate (IP6). Previous reports indicate that high inositol polyphosphates (IP4 and IP6) are involved in clathrin-coated vesicular recycling. In this study, we examined the effects of IP4 and IP6 on spontaneous transmitter release in the form of miniature endplate potentials (MEPP) and on enhanced vesicular recycling by high K+ at frog motor nerve endings. In resting conditions, IP4 and IP6 delivered intracellularly via liposomes, caused concentration-dependent increases in MEPP frequency and amplitude. Pretreatment with the protein kinase A (PKA) inhibitor H-89 or KT 5720 reduced the IP4-mediated MEPP frequency increase by 60% and abolished the IP6-mediated MEPP frequency increases as well as the enhancement in MEPP amplitude. Pretreatment with antibodies against phosphatidylinositol 3-kinase (PI 3-K), enzyme also associated with clathrin-coated vesicular recycling, did not alter the IP4 and IP6-mediated MEPP frequency increases, but reduced the MEPP amplitude increase by 50%. In our previous reports, IP3, but not other second messengers releasing Ca2+ from internal Ca2+ stores, is able to enhance the MEPP amplitude. In order to dissociate the effect of Ca2+ release vs. metabolism to IP4 and IP 6, we evaluated the effects of 3-deoxy-3-fluoro-inositol 1,4,5-trisphosphate (3F-IP3), which is not converted to IP 4 or IP6. 3F-IP3 produced an increase then decrease in MEPP frequency and a decrease in MEPP amplitude. In elevated vesicle recycling induced by high K+-Ringer solution, IP4 and IP6 have similar effects, except decreasing MEPP frequency at a higher concentration (10-4 M). We conclude that (1) high inositol polyphosphates may represent a link between IP3 and cAMP pathways; (2) the IP3-induced increase of MEPP amplitude is likely to be due to its high inositol metabolites; (3) PI 3-K is not involved in the IP 4 and IP6-mediated MEPP frequency increases, but may be involved in MEPP size.

An investigation into the influence of phytates on the in situ absorption of amino acids (lysine, glutamate and leucine) and glucose from the intestinal lumen of 3-week-old chickens was carried out. Birds were anaesthetised and the intestines exteriorised. Uptake of 5 mM of each nutrient over a 4-min period was measured in the presence of four phytate concentrations (0, 50, 250 and 500 mM). Five birds were used for each nutrient at each concentration of phytate tested. Leucine uptake decreased linearly (P < 0.001) and that of glutamate showed a tendency to decrease (P = 0.055) as the phytate concentration increased. Absorption of lysine and glucose were unaffected by the presence of phytate. In conclusion, phytate in the small intestinal lumen exerted a depressive effect on the absorption of specific free amino acids from the lumen. Its depressive effect was greatest for leucine followed by glutamate, and phytate had little effect on the absorption of lysine.

In the nervous system galanin primarily displays a modulatory role. The galaninergic system consists of a number of bioactive peptides with a highly plastic expression pattern and three different receptors. The lack of receptor subtype selective ligands and antibodies have severely hampered the charac-terization of this system. Therefore, most of the knowledge has been drawn from experiments with transgenic animals, which has given some major conclusions, despite the compensatory effects seen in several animal studies. Therefore, the production of subtype selective ligands is of great importance to delineate the galanin system and slowly experimental data from receptor subtype selective ligand trials is emerging. This thesis aims at studying galanin receptor-ligand interactions and to increase and improve the utilized tools in the galanin research field, espe-cially the development of novel galanin receptor subtype selective ligands. Paper I demonstrates the potential to N-terminally extend galanin ana-logues and the successful development of a galanin receptor 2 (GalR2) selec-tive ligand. In addition, a cell line stably expressing galanin receptor 3 (GalR3) was developed, to improve and simplify future evaluations of sub-type selective galanin ligands. Paper II measures the affinities of M617 and M871 to GalR3 and demon-strates that M871 preferentially binds GalR2. Furthermore, the relatively high affinity of M617 was evaluated by assessing the contribution in recep-tor interaction of individual amino acid residues in the C-terminal part of the M617. In conclusion, this thesis has provided a novel design strategy for galanin receptor ligands and increased the understanding of ligand interactions with the GalR3. Furthermore, M1145 has together with new analogues proven to be highly GalR2 specific, holding promises to future delineation of the galaninergic system as a therapeutic target.

Introduktion: Polycystisk ovarial syndrom (PCOS) är ett vanligt förekommande endokrinsjukdom som oftast drabbar kvinnor i fertil ålder. För att bli bekräftad diagnosen PCOS så måstetre kritetrier gällande kvinnans hälsåtillstånd uppfyllas; ovulationsstörningar, hyperandrogenism och polycystiska ovarial morfologi. Förutom de nämnda kriterierna såverkar insulinresistens (IR) ha en viktig roll för PCOS patofysiologi. Därför har ämnen sompåverkar insulinkänsligheten såsom myo-inositol (MI) hamnat i större fokus som potentiellbehandling av PCOS. Därav har syftet med detta arbete varit att undersöka om MI förbättrarovulation och fertilitet för kvinnor med diagnosen PCOS. Metod: För att genomföra detta litteraturstudie har vetenskapliga artiklar sökts via databasen PuBMED med sökorden ”PCOS and myoinositol on infertility” och ”myoinositol for PCOS”. Totalt valdes fem originalstudier vilka svarade mot utvalda urvalskriterier, därefteranalyserades och sammanställdes de fem valda studierna vidare för att undersöka hur MItillskott påverkar ovulationsstörningar och infertilitet hos kvinnor med PCOS. Resultat: Majoriteten av de valda studierna visade en statistisk signifikant förbättring påkvinnans ovulationsstörning, vilket ledde till förbättrad fertilitet och därmed uppnåddesgraviditet hos många deltagare i studierna. Slutsats: Sammanfattningsvis visar MI tillskaott vara en bra behandling motovulationsstörningar och infertilitet hos kvinnor med PCOS. Dock bör fler studier genomförasdär hänsyn till hur MI tillskott påverkar androgennivåer för att förbättra ovulationsstörning ochinfertilitet vid PCOS tas in.