Fler unga människor än någonsin är idag överviktiga och stress och stressrelaterade symptom drabbar idag allt fler unga. Därför är det viktigt att unga människor får kunskaper om hur de på bästa sätt kan ta hand om sig själva.  Syftet med detta arbete är att undersöka hur elever som läser gymnasiets kurs Idrott och hälsa A ser på den hälsoundervisning de får. Detta är relevant för alla som arbetar som idrott och hälsa lärare för att kunna hitta en jämkning mellan kursplan och elevernas tankar och förkunskaper.  Genom intervjuer och fokusintervjuer kom jag fram till att eleverna vill lära sig mer om stress och hur man hanterar stress samt om kost. Eleverna tycker däremot att idrottsläraren inte är rätt person att lära ut kunskaper om tobak.

<p>Fler unga människor än någonsin är idag överviktiga och stress och stressrelaterade symptom drabbar idag allt fler unga. Därför är det viktigt att unga människor får kunskaper om hur de på bästa sätt kan ta hand om sig själva.</p><p> Syftet med detta arbete är att undersöka hur elever som läser gymnasiets kurs Idrott och hälsa A ser på den hälsoundervisning de får.</p><p>Detta är relevant för alla som arbetar som idrott och hälsa lärare för att kunna hitta en jämkning mellan kursplan och elevernas tankar och förkunskaper.</p><p> Genom intervjuer och fokusintervjuer kom jag fram till att eleverna vill lära sig mer om stress och hur man hanterar stress samt om kost. Eleverna tycker däremot att idrottsläraren inte är rätt person att lära ut kunskaper om tobak.</p>

Glucose uptake occurs in skeletal muscle under basal conditions, and increases in response to stimuli such as insulin and exercise. Exercise is known to increase blood flow, and it appears that insulin has similar hemodynamic effects, including increased blood flow and capillary recruitment, which can modify the amount of glucose uptake occurring under each condition. Here we study factors affecting both basal and stimulated myocyte glucose uptake, with a particular focus on vasoactive agents. Insulin stimulates the release of endothelin-1 (ET-1), a potent vasoconstrictor, from endothelial cells in culture. As yet it is unknown whether ET-1 is a type A (causing nutritive perfusion) or a type B (non-nutritive) vasoconstrictor, so here we use the pump-perfused rat hindlimb to characterize the distribution effects of ET-1. We show that ET-1 causes a type A vasoconstriction, stimulating basal metabolism at low doses, while at high doses the distribution of flow changes to become non-nutritive, inhibitory to metabolism. As a general vasodilator prevents both metabolic and hemodynamic effects, the effects on metabolism are due to the redistribution of flow. These redistribution effects are confirmed by the ability of high dose ET-1 to decrease aerobic tension development in the contracting hindlimb, and by the ability of low dose ET-1 to increase the interstitial glucose concentration. Given this understanding of the effects of ET-1 alone, we can investigate the interactions between ET-1 and insulin. In the perfused rat hindlimb, insulin has not been observed to have any vasodilatory effect, whereas here for the first time insulin appears to have vasodilator-like actions against ET-1 mediated vasoconstriction. Also, the redistribution of flow by ET-1 does not appear to alter the metabolic effect of insulin to cause glucose uptake at either dose of ET-1 used. Nitric Oxide (NO) is thought to be the mechanism by which insulin causes vasodilation in muscle. A previous study has shown that methacholine (MC), by increasing NO, was able to augment insulin-mediated glucose uptake and capillary recruitment, while other NO donors were unable to do so. Here we show that, at the dose used to increase glucose uptake in the previous study, MC has only a vasodilatory effect, and no direct effect on glucose uptake, in the perfused rat hindlimb. At higher doses, an effect on glucose uptake can be observed. This means that the increase in capillary recruitment by MC was responsible for the elevated insulin-mediated glucose uptake, and there was no direct effect of MC on glucose uptake. A recent publication suggested that the Na+-D-glucose cotransporter (SGLT1) was essential for insulin-mediated glucose uptake, although not required for basal glucose uptake. The implications of this detract from our proposed role of blood flow redistribution in insulin action. In attempting to reproduce these results in the perfused rat hindlimb we found that SGLT1 is not required for insulin-mediated glucose uptake, and confirmed this using a low sodium buffer, which would also inhibit the transporter. We conclude that SGLT1 is not required for insulin-mediated glucose uptake. Our results therefore suggest that complex interactions are involved in insulin action, some of which involve hemodynamic actions that are capable of altering insulin-mediated glucose uptake, and others in which insulin itself can limit the action of other vasomodulators, such as ET-1. It is apparent, however that SGLT1 in the endothelium may not be necessary for the metabolic effects of insulin, and that blood flow distribution, or capillary recruitment is therefore of great importance in delivering glucose to myocytes.

<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>The aim of this study was to examine the expression of Thrombomodulin, Tissue Factor, Tissue Factor Pathway Inhibitor and Endothelial Protein C Receptor in placenta throughout the three phases of the third trimester in the normal placenta and in IUGR placenta from full term.</p><p>Twenty-five normal placenta samples and twenty-five IUGR placenta samples were obtained and each sample was stained by immunohistochemistry using monoclonal antibodies. Each antibody was optimised for antigen retrieval method and for optimal dilution, before been applied to the test tissue.</p><p>The results showed that each of the antibodies mentioned was expressed in normal placenta and in IUGR placenta.</p><p>No significant difference could be established concerning the expression of each antibody mentioned between normal and IUGR placenta.</p>

Each cell division, the nuclear DNA must be replicated efficiently and with high accuracy to avoid mutations which can have an effect on cell function. There are three replicative DNA polymerases essential for the synthesis of DNA during replication in eukaryotic cells. DNA polymerase α (Pol α) synthesize short primers required for DNA polymerase δ (Pol δ) and DNA polymerase ε (Pol ε) to carry out the bulk synthesis. The role of Pol δ and Pol ε at the replication fork has been unclear. The aim of this thesis was to examine what role Pol ε has at the replication fork, compare the biochemical properties of Pol δ and Pol ε, and to study the function of the second largest and essential subunit of Pol ε, Dpb2. To identify where Pol ε replicates DNA in vivo, a strategy was taken where the active site of Pol ε was altered to create a mutator polymerase leaving a unique error-signature. A series of mutant pol ε proteins were purified and analyzed for enzyme activity and fidelity of DNA synthesis. Two mutants, M644F and M644G, exhibited an increased mutation rate and close to normal polymerase activity. One of these, the M644G gave rise to a specific increase of mismatch mutations resulting from T-dTMP mis-pairing during DNA synthesis in vitro. The M644G mutant was introduced in yeast strains carrying a reporter gene, URA3, on either side of an origin in different orientations. Mutations which inactivated the URA3 gene in the M644G mutant strains were analyzed. A strand specific signature was found demonstrating that Pol ε participates in the synthesis of the leading strand. Pol δ and Pol ε are both stimulated by the processivity clamp, PCNA, in in vitro replication assays. To clarify any differences they were challenged side by side in biochemical assays. Pol ε was found to require that single-stranded template (ssDNA) was entirely coated with RPA, whereas Pol δ was much less sensitive to uncoated ssDNA. The processivity of Pol δ was stimulated to a much higher degree by PCNA than of Pol ε. In presence of PCNA the processivity of Pol δ and Pol ε was comparable. In contrast, Pol ε was approximately four times slower than Pol δ when replicating a single-primed circular template in the presence of all accessory proteins and an excess of polymerase. The biochemical characterization of the system suggests that Pol ε and Pol δ are loaded onto the PCNA-primer-ternary complex by separate mechanisms. A model is proposed where the loading of Pol ε onto the leading strand is independent of the PCNA interaction motif which is required by enzymes acting on the lagging strand. The essential gene DPB2 encodes for the second largest subunit of Pol ε. We carried out a genetic screen in S.cerevisiae and isolated a lethal mutant allele of dpb2 (dpb2-200). When over-expressed together with the remaining three subunits of Polε, Pol2, Dpb3 and Dpb4, the dpb2-201 did not copurify. The biochemical property of Pol2/Dpb3/Dpb4 complex was compared with wild-type four-subunit Pol ε (Pol2/Dpb2/Dpb3/Dpb4) and a Pol2/Dpb2 complex in replication assays. The absence of Dpb2 in the complex did not significantly affect the specific activity or the processivity, but gave a slightly reduced efficiency in holoenzyme assays when compared to wild-type four-subunit Pol ε. We propose that Dpb2 is not essential for the enzyme activity of Pol ε.

The aim of this study was to examine the expression of Thrombomodulin, Tissue Factor, Tissue Factor Pathway Inhibitor and Endothelial Protein C Receptor in placenta throughout the three phases of the third trimester in the normal placenta and in IUGR placenta from full term. Twenty-five normal placenta samples and twenty-five IUGR placenta samples were obtained and each sample was stained by immunohistochemistry using monoclonal antibodies. Each antibody was optimised for antigen retrieval method and for optimal dilution, before been applied to the test tissue. The results showed that each of the antibodies mentioned was expressed in normal placenta and in IUGR placenta. No significant difference could be established concerning the expression of each antibody mentioned between normal and IUGR placenta.

Several methods for the fractionation of transfer RNA have been investigated. These include countercurrent distribution, ion exchange chromatography, chemical methods utilizing the oxidation of transfer RNA with periodate, and hydrogen bonding methods. The effect of temperature on the ion exchange fractionation characteristics of yeast transfer RNA has been studied using the anion exchanger diethylaminoethyl-cellulose (DUE--cellulose).

Several methods for the fractionation of transfer RNA have been investigated. These include countercurrent distribution, ion exchange chromatography, chemical methods utilizing the oxidation of transfer RNA with periodate, and hydrogen bonding methods. The effect of temperature on the ion exchange fractionation characteristics of yeast transfer RNA has been studied using the anion exchanger diethylaminoethyl-cellulose (DUE--cellulose).

Restricted Item. Print thesis available in the University of Auckland Library or available through Inter-Library Loan. 1. Antitumour agents from many different chemical classes including acridines, have been shown to induce topoisomerase II-associated DNA breaks both in cultured mammalian cells and in vitro in the presence of purified mammalian DNA topoisomerase II. However, mechanisms linking these protein-associated DNA breaks with drug cytotoxicity are poorly understood. Whether this type of DNA damage is responsible for drug cytotoxicty was investigated using a number of acridine derivatives belonging to the sane chemical class but with varying potency in vivo. The modulation of topoisomerase II-mediated DNA breaks in whole L1210 cells, isolated nuclei and nuclear extract systems correlated with drug-induced cytotoxicity, at least for the drugs belonging to the amsacrine lineage. In contrast, there was no direct relationship between the inhibition of topoisomerase II strand-passing activity and the cytotoxic action of topoisomerase II-specific anticancer drugs. It was concluded that drug-induced formation and stabilization of the topoisomerase II-DNA cleavable complex rather than inhibition of its formation or of strand-passing functions, is responsible for DNA strand breakage and cell death caused by antitumour acridines such as amsacrine. 2. Two tissue culture systems (CHO-AA8 cells or P815 cell cycle mutants) were used as models to study the involvement of topoisomerase II in the resistance of non-cycling cells to amsacrine. Plateau-phase CHO-AA8 cells with a GO/Gl DNA content are resistant, to amsacrine and contain fewer DNA breaks than log-phase cells after drug treatment. (Robbie et al., 1988). The observations of Robbie et al. (1988) were further investigated in this thesis. The phage P4 DNA unknotting activity in CHO-AA8 cell nuclear extracts decreased 2-fold when the cells entered plateau-phase, but there was no difference in the sensitivity of unknotting to amsacrine between log- and plateau-phase nuclear extracts. Furthermore, drug stimulation of protein-DNA complex formation was similar in whole cells, isolated nuclei and nuclear extracts from either log- or plateau-phase cells. However, stimulation of complex formation in cells, nuclei or nuclear extracts was approximately 4-fo1d lower in plateau-phase than in log-phase. These results suggested that drug-enzyme interaction was altered in plateau-phase and there was a good correlation between the proliferative state of cells, amsacrine sensitivity DNA breakage and topoisomerase II-DNA complex formation. The second system used to study the effect of amsacrine was the cold-sensitive (proliferating at 39.5°C; reversibly arrested in Gl-phase at 33°C) cell-cycle mutant 21-Fb of the murine mastocytoma cell line, P815- The sensitivity of arrested 21-FB cells to amsacrine decreased –less than two-fold in cell survival experiments when compared to proliferating cells. In contrast, DNA breakage and stimulation of protein-DNA complex formation in intact cells, lysed cells or isolated nuclei was reduced approximately 10-fold in amsacrine-treated arrested cells and DNA-topoisomerase II activity in arrested cells was only 5% of the activity in proliferating cells. Thus in contrast to the CHO-AA8 cell system, there was no correlation between cell survival and DNA damage or DNA topisomerase II activity in drug treated 21-Fb cells. The exact reasons for the differences in amsacrine sensitivity in growth arrested CHO and 21-Fb cells were not resolved. However, it was concluded that a complex relationship exists between amsacrine-induced DNA breakage, topoisomerase II-DNA complex formation, topoisomerase activity and drug cytotoxicity. Moreover, a very complex set of parameters can influence drug-induced topoisomerase II-mediated lesions and cytotoxicity in different cells or even a single type of cell under different growth conditions. 3. Extracts of K21 murine mastocytoma cells were found to contain a factor that, enhances formation of amsacrine-induced topoisomerase II-DNA complexes (PDC) when added to isolated K21 nuclei. The PDC enhancing activity was reduced in extracts from 2 or 6 h cycloheximide- or cordycepin-treated cells, implying that continuous protein synthesis is required to maintain the factor. Preliminary characterization showed that the factor was heat labile and proteinase-sensitive suggesting the factor was a labile protein which was distinct from the two known classes of topoisomerase. The protein factor was present in at least four other cell lines and was substantially reduced in cells induced into a G1 state by temperature arrest or serum deprivation, Human Jurkat cells selected for resistance to amsacrine and displaying cross-resistance to other topoisomerase II-targeted drugs also exhibited significantly reduced PDC enhancing activity. These results suggested a contributing role for the PDC enhancing factor in mediation of drug resistance. Fractionation of mouse mastocytoma cell cytoplasmic extracts by salt precipitation, DEAE-cellulose chromatography and SDS-PAGE resulted in a 3571-fold purification of the PDC enhancing activity. The PDC enhancing activity was shown to reside in a 70 kDa protein kinase with specificity for a casein kinase II substrate and sensitive to heparin and anti-casein kinase II antiserum. This appears to be the first direct evidence of a protein factor that modulates amsacrine induced topoisomerase II action possibly by phosphorylating topoisomerase II or proteins associated with topoisomerase II.