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Det har forskats mycket på olika signalvägar i det mänskliga genomet, trotts detta finns det många frågetecken som kvarstår. Denna uppsats undersöker några av dem. Syfte: Undersöka förändringar i genuttryck och mRNA-nivåer för hypertrofi- (MRF4) translations- (5.8S & 18S) och proteolysreglerande gener (MuRF1 & GDF-8) efter en 10 veckor lång styrketräningsperiod hos kvinnor och män. Frågeställningar: (1) Finns det en förändring i total mängd RNA före och efter en 10 veckors styrketräningsintervention. (2) Finns det en förändring i uttryck av MRF4, 5.8S, 18S, MuRF1 samt GDF-8 efter en 10 veckors styrketräningsintervention. (3) Finns det en könsskillnad i förändringen av total mängd RNA samt aktivering av MRF4, 5.8S, 18S, MuRF1 och GDF-8 efter en 10 veckors styrketräningsintervention. Metod: Urvalet för analysen bestod av 16 otränade försökspersoner varav 8 var män och 8 var kvinnor. Försökspersonerna utförde unilateral styrketräning av nedre extremiteten under 10 veckor, under 2 av dessa veckor utfördes ocklusionsträning.  Träningsperiodiseringen var vågformig (70-90% av 1RM, 5-12 rep, 3 ggr/vecka). Muskelbiopsier togs i det arbetande benet före träningsperiodens start samt 3-7 dagar efter träningsperiodens avslut. Genuttryck analyserades med qPCR. Resultat: Det fanns ingen signifikant skillnad i förändring mellan män och kvinnors totala RNA eller genuttryck. Total RNA ökade signifikant (p<0,01) med 19,2 %. Kvinnorna hade en signifikant ökning (P<0,05) av RNA på 27,6 % medan männen hade en signifikant ökning (p<0,05) på 14 %. MRF4 hade en signifikant (P>0,05) procentuell ökning i genuttryck med 55,7 % och kvinnor för sig hade en signifikant (P>0,05) ökning på 64 %. GDF-8 ökade signifikant (P>0,05) med 55,5 % medan GAPDH ökade signifikant (P>0,05) för båda könen tillsammans med 70,6 % och för män med 87,8 %. MuRF1 och 5.8S hade inga signifikanta förändringar i genuttryck. Slutsats: Det verkar som att både män och kvinnor får en liknande procentuell förändring av total RNA och mRNA genuttryck 3-7 dagar efter en 10 veckors hypertrofistyrd styrketräningsperiod. För att mäta genuttryck av translationsgenen MRF4 verkar 3-7 dagar efter en 10 veckors styrketräningsperiod vara en tidpunkt då det fortfarande pågår hypertrofi av skelettmuskulaturen.  Av de proteolysreglerande generna GDF-8 och MuRF1 sågs en uppreglering av GDF-8 vilket skulle kunna vara ett tecken på att hypertrofin börjar hämmas. Ett oväntat fynd var att GAPDH visade sig vara olämplig som kontrollgen vid en styrketräningsintervention på 10 veckor och att 18S var väldigt stabil. Detta kan betyda att GAPDH inte skall användas vid längre styrketräningsinterventioner. / There have been much research on signaling pathways in the human genome, but there still remain many questions. This paper examines some of them. Aim: Investigate changes in gene expression and mRNA levels of hypertrophy (MRF4), translation (5.8S & 18S) and proteolysis regulating genes (GDF-8) after a 10-week strength training period in men and women. Research questions: (1) Is there a change in the total amount of RNA before and after a 10-week strength training intervention. (2) Is there a change in the expression of MRF4, 5.8S, 18S, Murf1 and GDF-8 after 10 weeks of strength training. (3) Is there a gender difference in the change of total RNA and the expression of MRF4, 5.8S, Murf1 and GDF-8 after a 10-week long strength training intervention. Method: The sample for analysis consisted of 16 untrained subjects, of whom 8 were men and 8 were women. The subjects performed unilateral resistance training of lower extremities for 10 weeks, during two of these weeks blood flow restriction training were performed. The training was undulating (70-90% of 1RM, 5-12 cord, 3 times / week). Muscle biopsies were taken from the working leg before the start and 3-7 days after the training period. Gene expression was analyzed by qPCR. Results: There was no significant gender difference in total RNA or gene expression. Total RNA was significantly increased (p <0.01) with 19.2 %. The women had a significant increase (P <0.05) of RNA at 27.6 %, while the men had a significant increase (p <0.05) at 14 %. MRF4 had a significant (P> 0.05) percentage increase in gene expression by 55.7 %, and women had a significant (P> 0.05) increase of 64 %. GDF-8 increased significantly (P> 0.05) with 55.5 %, while GAPDH increased significantly (P> 0.05) for both sexes with 70.6 % and for men with 87.8 %. Murf1 and 5.8S had no significant changes in gene expression. Conclusions: It seems that both men and women experience a similar percentage difference of total RNA and mRNA gene expression 3-7 days after a 10 weeks long strength training period. To measure the gene expression of MRF4 3-7 days after a 10-week weight-training period seems to be a time when there still is a anabolic responses in the skeletal muscle. Of the proteolysis regulating genes GDF-8 and Murf1 there was an upregulation of GDF-8, which could be a sign that the inhibition of hypertrophy started. An unexpected finding is that GAPDH was found to be unsuitable as a control gene at a strength training intervention at 10 weeks and rRNA 18S was very stable, which could mean that GAPDH should not be used as control gene in longer strength training studies.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / Purpose: Breast cancer is one of the most frequent and fatal diseases women all around the globe are challenged with today. In women, breast cancer has the highest mortality rate of all cancers and the incidence rate is on the increase. It is estimated that by the year 2025, 19.3 million women will become a victim of this grave health problem. This disease is a result of the formation of malignant tumours caused by genetic alterations that are involved in the proliferation of cells, cellular differentiation and the disturbance in homeostasis which subsequently leads to the abnormal multiplication and growth of cells. Breast cancer is considered a multifactorial disease with various risk factors such as age, radiation exposure, hormone therapy, oral contraceptives, dietary factors, environmental exposure and genetic predispositions. Breast cancers can be subdivided and classified based on their cellular surface receptors such as Estrogen Receptors, Progesterone Receptors and Human Epidermal Growth Factor Receptor 2. Of the various subtypes, the triple-negative breast cancer subtype which is negative for all 3 surface receptors and presents as the most aggressive form of breast cancer with a poor prognosis. Between 10-20% of all breast cancer cases are classified as triple-negative breast cancer. Due to the hormonal status of triple-negative breast cancer, treatment options are limited and thus of great concern. Chemotherapy remains the most common treatment modality, but prognosis is poor with relapse within years ultimately leading to poor survival outcome. Due to this lack of effective treatment plans, an alternative treatment with minimal side effects and better survival remains an imperative area to explore. A wide scope of literature highlights red palm oil and its health benefits, with its growth inhibitory potential drawing great attention. Red palm oil, extracted from the Elaeis guineensis palm tree is red in colour due to the abundance of carotenoids, tocotrienols and tocopherols found in the oil. Various compounds make up the oil such as lycopene, carotenes, vitamin E and coenzyme Q10. Most studies have researched the effects of vitamin E extracted from the oil as a contributor to its growth inhibitory activity. This study focuses on the effects of the commercial red palm oil as a whole with all its compounds on the proliferation of breast cancer cells as well as the effect it has on various genes associated with breast cancer. Method: This study investigated the effect of red palm oil concentrations (1, 10, 100, 500 and 1000 μg/ml) on breast cancer cells—MCF-7 and MDA-MB-231 with comparison to a non-cancerous cell line—MCF-12A for 24-, 48- and 72-hour treatment periods. The parameter investigated was cell proliferation through the CCK-8 cell proliferation assay and the morphology following red palm oil treatment was observed and captured. Additionally, this study also investigated the effect of red palm oil on the expression of Human Mammaglobin (hMAM) and Maspin genes through the PCR assay and results visualised through agarose gel electrophoresis. Data was statistically analysed using GraphPad version 6.0 software. Results: Following treatment of red palm oil, no apparent changes in the cell morphology was observed despite using variable treatment concentrations over variable times for MCF-7, MDA-MB-231 and MCF-12A cells relative to their respective controls. Immortalised MCF-12A cells showed a significant increase in proliferation with the varying treatment concentrations, but more prominently with the highest concentration at 24, 48 and 72 hours. MCF-7 cells showed significant decreases at 24 and 72 hours. Decreased proliferation was observed at all dosages used, particularly at 10, 100, and 500 μg/ml. Furthermore, MDA-MB-231 cells demonstrated a gradual increase in cell proliferation for the 3 selected time periods in the varying concentrations. Additionally, red palm oil did not alter the gene expression of Maspin at any of the varying treatments for MDA-MB-231 nor MCF-7 cells. However, changes in hMAM gene expression were observed at treatment concentration of 100 μg/ml in MDA-MB-231 cells that were incubated for 24 and 48 hours. However, the hMAM expression was not affected in treated MCF-7 cells. Conclusion: Red palm oil, as an alternative dietary oil, seems to have potential growth inhibitory properties as demonstrated by the change in the cell proliferation of the MCF-7 cells. Literature show that various individual compounds extracted from red palm oil have anti-proliferative and inhibitory effects on breast cancer cells making them good candidates for therapy. However, this study concludes that red palm oil as a whole component would not be a suitable therapeutic agent for highly aggressive triple-negative breast cancer.