<p>Prostate cancer is common in the Western society and current treatments are often associated with side effects, therefore improved therapeutic strategies are desired. 2-methoxyestradiol (2-ME), an endogenous metabolite of estradiol-17β inhibits tumor growth <i>in vivo</i> as it prevents angiogenesis. 2-ME has also direct cytotoxic effects on tumor cells. In this study, we have investigated the potential use of PET to record effects 2-ME on prostate cancer cell (PC3) aggregates. The anti-proliferative and pro-apoptotic effects of 2-ME on PC3 cell aggregates <i>in vitro</i> were correlated with the uptake of deoxy-D-glucose, FMAU and choline labeled with ¹⁸F, ¹¹C or ³H. 2-ME clearly reduced growth of PC3 aggregates and induced apoptosis in a dose-dependent manner. However, the PET tracers failed to record the cytotoxicity of 2-ME on PC3 aggregates. </p><p>Further, the signaling events responsible for 2-ME induced prostate cancer cell death were investigated. We found that Smad7, previously implicated in TGF-β-induced responses, is required for 2-ME-induced p38 MAPK activation and subsequent apoptosis in PC-3U cells, as shown by the use of antisense or siRNA techniques and a specific inhibitor of p38 MAPK (SB203580). Interestingly, Smad7 also regulated the expression of the pro-apoptotic Bim protein. </p><p>Shb is a Src Homology 2 domain adapter protein with pro-apoptotic effects. PC3 clones overexpressing Shb exhibited increased rates of apoptosis, both in the presence or absence of 2-ME, as they failed to activate survival mechanisms through ERK and Akt in response to 2-ME. Notably, Shb cells displayed increased activity of the pro-apoptotic kinase c-Abl. Pre-treatment with SB203580 or c-Abl (STI-571) inhibitors completely blocked the apoptotic response to 2-ME. </p><p>In conclusion, Smad7 and Shb appear to be crucial for 2-ME-induced PC3 cell apoptosis via their activation of p38 MAPK and c-Abl. Future therapies exploring these pathways can be envisaged as treatment of prostate cancer.</p>

Prostate cancer is common in the Western society and current treatments are often associated with side effects, therefore improved therapeutic strategies are desired. 2-methoxyestradiol (2-ME), an endogenous metabolite of estradiol-17β inhibits tumor growth in vivo as it prevents angiogenesis. 2-ME has also direct cytotoxic effects on tumor cells. In this study, we have investigated the potential use of PET to record effects 2-ME on prostate cancer cell (PC3) aggregates. The anti-proliferative and pro-apoptotic effects of 2-ME on PC3 cell aggregates in vitro were correlated with the uptake of deoxy-D-glucose, FMAU and choline labeled with 18F, 11C or 3H. 2-ME clearly reduced growth of PC3 aggregates and induced apoptosis in a dose-dependent manner. However, the PET tracers failed to record the cytotoxicity of 2-ME on PC3 aggregates. Further, the signaling events responsible for 2-ME induced prostate cancer cell death were investigated. We found that Smad7, previously implicated in TGF-β-induced responses, is required for 2-ME-induced p38 MAPK activation and subsequent apoptosis in PC-3U cells, as shown by the use of antisense or siRNA techniques and a specific inhibitor of p38 MAPK (SB203580). Interestingly, Smad7 also regulated the expression of the pro-apoptotic Bim protein. Shb is a Src Homology 2 domain adapter protein with pro-apoptotic effects. PC3 clones overexpressing Shb exhibited increased rates of apoptosis, both in the presence or absence of 2-ME, as they failed to activate survival mechanisms through ERK and Akt in response to 2-ME. Notably, Shb cells displayed increased activity of the pro-apoptotic kinase c-Abl. Pre-treatment with SB203580 or c-Abl (STI-571) inhibitors completely blocked the apoptotic response to 2-ME. In conclusion, Smad7 and Shb appear to be crucial for 2-ME-induced PC3 cell apoptosis via their activation of p38 MAPK and c-Abl. Future therapies exploring these pathways can be envisaged as treatment of prostate cancer.

Är de svenska storbankernas presentation av finansiella nyckeltal jämförbara? Det är frågan jag ställt mig i syfte att skapa förståelse för omfattningen av skillnaden mellan bankernas jämförande information via finansiella nyckeltal. Analysen har utförts genom en fallstudie, där ett systemsynsätt tillämpats, och har visat att vare sig bankernas presentation av finansiella nyckeltal, eller teorier kring nyckeltal, kan anses vara jämförbara. Stora skillnader existerar, både kring vilka nyckeltal som presenteras, vad de kallas och hur de beräknas. Finansiella nyckeltal kan således inte anses vara användbara som analysverktyg, eller bidra till den inre marknadens funktion. Resultatet av studien baserar sig på analyser av bankernas presenterade finansiella nyckeltal, resultat- och balansräkningar, ägarstrukturer och börskurser. Grunddata har hämtats från bankernas årsredovisningar under perioden 2004 – 2008, från Finansinspektionen och Stockholmsbörsen. Jag har även intervjuat representanter från bankerna och Finansinspektionen, en analytiker och en ekonomijournalist. Den övergripande bilden och djupa förståelsen har jag fått av Jacob Palmstierna, som haft godheten att dela med sig av sin fyrtioåriga erfarenhet och ovärderliga kunskap om den svenska bankbranschen. Uppsatsen innefattar även metodutveckling i form av en konceptuell omvärldsmodell.

Many rural communities in South Africa do not have running water or electricity. The fetching and heating of water is therefore a time consuming and expensive daily ritual. The use of energy sources such as wood or coal are not readily available and cause environmental pollution. Although solar water heaters are commonly available in South Africa, they are very seldom used in rural areas. Whilst this can mostly be attributed to a high system cost, current designs also do not cater for specific rural problems such as the transporting or purification of water. A prototype model, designed with such an approach in mind, has already been constructed by TEMM International (Pty.) Ltd. The Solar Heat Barrow (SHB) was developed in the 1992 to 2003 period with the intention of combining a number of functions at low cost. Specific requirements were: low cost, a suitable design and materials for manufacture in large volume, sufficiently durable taking into account the harsh conditions of use. suitability for cases where no in-house piped water supply was available, the use of appropriate technology and the improvement of quality of life. The unit combines the absorption of solar radiation, the heating of a relatively small volume of water, the transport of the water from the point of supply and the storage of the hot water until it is used. Untreated water sources such as surface waters (streams, rivers, lakes, etc.) or unprotected open wells are the vehicles for waterborne bacterial diseases such as cholera and typhoid fevers. In the case where water is collected from these sources, the SHB has a build-in Purification Dispenser that purifies the water in the collector against waterborne bacterial diseases. Certain research questions need to be answered. They will be answered by demonstrating the SHB in two communities where no in-house piped water supply is available and by establishing the socio-economic response of the users. The research questions are as follow: What are the responses of the users concerning the SHB, in comparison to those of a control group, regarding its operation, durability, utility and satisfaction of needs? What is the daily use of hot water and the reduction in energy use and cost? To what extent will the target community purchase the SHB at the full or subsidised commercial price? Is there a business case that can be developed for the large scale production, marketing. financing and Small, Medium and Micro Enterprises (SMME) development of the SHB? It was decided to choose a community in the Valley of Thousand Hills in KwaZulu Natal as the demonstration site. The name of the community is Mabedlane. It is a remote rural area 20 km's north of Botha's Hill and is situated along the Umgeni River. The community is dependent on the river for domestic water. Most families are headed by women. The area has low levels of infrastructure, poor roads, a high unemployment rate and poor health facilities. The first survey, which was conducted before the test period started, showed a very positive response from potential users. From 112 questionnaires that were given to the people of Mabedlane, all indicated that they were interested in a product that can transport and heat water for domestic use. The socio-economic study has shown that the price per unit needs to be adapted as 85% of the people who participated in the survey indicated that they would only pay less than R100 for the product. 15% indicated that they would pay between RlOO and R200. From the second and third questionnaires it was clear that the users were satisfied with the heating performances of the SHB. The community was very interested in purchasing a SHB. They have realised that a SHB will improve their standard of living and regard it as a necessity in their day to day activities. It is apparent that people, who will benefit from a SHB most, are those who will not be able to pay the full retail price. Therefore, new business strategies have to be researched when implementing the SHB to the target market in South Africa. The idea of considering ways to sponsorlfund the SHB must also be investigated. / Thesis (M.Ing. (Mechanical Engineering))--North-West University, Potchefstroom Campus, 2004.

Many rural communities in South Africa do not have running water or electricity. The fetching and heating of water is therefore a time consuming and expensive daily ritual. The use of energy sources such as wood or coal are not readily available and cause environmental pollution. Although solar water heaters are commonly available in South Africa, they are very seldom used in rural areas. Whilst this can mostly be attributed to a high system cost, current designs also do not cater for specific rural problems such as the transporting or purification of water. A prototype model, designed with such an approach in mind, has already been constructed by TEMM International (Pty.) Ltd. The Solar Heat Barrow (SHB) was developed in the 1992 to 2003 period with the intention of combining a number of functions at low cost. Specific requirements were: low cost, a suitable design and materials for manufacture in large volume, sufficiently durable taking into account the harsh conditions of use. suitability for cases where no in-house piped water supply was available, the use of appropriate technology and the improvement of quality of life. The unit combines the absorption of solar radiation, the heating of a relatively small volume of water, the transport of the water from the point of supply and the storage of the hot water until it is used. Untreated water sources such as surface waters (streams, rivers, lakes, etc.) or unprotected open wells are the vehicles for waterborne bacterial diseases such as cholera and typhoid fevers. In the case where water is collected from these sources, the SHB has a build-in Purification Dispenser that purifies the water in the collector against waterborne bacterial diseases. Certain research questions need to be answered. They will be answered by demonstrating the SHB in two communities where no in-house piped water supply is available and by establishing the socio-economic response of the users. The research questions are as follow: What are the responses of the users concerning the SHB, in comparison to those of a control group, regarding its operation, durability, utility and satisfaction of needs? What is the daily use of hot water and the reduction in energy use and cost? To what extent will the target community purchase the SHB at the full or subsidised commercial price? Is there a business case that can be developed for the large scale production, marketing. financing and Small, Medium and Micro Enterprises (SMME) development of the SHB? It was decided to choose a community in the Valley of Thousand Hills in KwaZulu Natal as the demonstration site. The name of the community is Mabedlane. It is a remote rural area 20 km's north of Botha's Hill and is situated along the Umgeni River. The community is dependent on the river for domestic water. Most families are headed by women. The area has low levels of infrastructure, poor roads, a high unemployment rate and poor health facilities. The first survey, which was conducted before the test period started, showed a very positive response from potential users. From 112 questionnaires that were given to the people of Mabedlane, all indicated that they were interested in a product that can transport and heat water for domestic use. The socio-economic study has shown that the price per unit needs to be adapted as 85% of the people who participated in the survey indicated that they would only pay less than R100 for the product. 15% indicated that they would pay between RlOO and R200. From the second and third questionnaires it was clear that the users were satisfied with the heating performances of the SHB. The community was very interested in purchasing a SHB. They have realised that a SHB will improve their standard of living and regard it as a necessity in their day to day activities. It is apparent that people, who will benefit from a SHB most, are those who will not be able to pay the full retail price. Therefore, new business strategies have to be researched when implementing the SHB to the target market in South Africa. The idea of considering ways to sponsorlfund the SHB must also be investigated. / Thesis (M.Ing. (Mechanical Engineering))--North-West University, Potchefstroom Campus, 2004.

<p>Protein tyrosine kinases play an important role in the regulation of various cellular processes such as</p><p>growth, differentiation and survival. GTK, a novel SRC-like cytoplasmic tyrosine kinase, was recently cloned from a mouse insulinoma cell line and the present work was conducted in order to find a biological function of GTK in insulin producing and neuronal cells. It was observed that kinase active GTK-mutants, expressed in RINm5F cells, transferred to the cell nucleus and increased the levels of the cell cycle regulatory protein p27^KIP1, reduced cell growth and stimulated glucagon mRNA expression. Furthermore, wild type GTK induces neurite outgrowth in the rat adrenal pheochromocytoma PC12 cell line, through activation of the RAP1-pathway, suggesting a role of GTK for cell differentiation. Studies using transgenic mice, expressing GTK under the control of the rat insulin 1 promoter, demonstrated a dual role of GTK for β-cell growth: Whereas GTK increases the β-cell mass and causes enhanced β-cell proliferation in response to partial pancreatectomy it also induced β-cell death in response to proinflammatory cytokines and impaired the glucose tolerance in mice treated with the β-cell toxin streptozotocin suggesting a possible role of GTK for β-cell destruction in Type 1 diabetes. We have also observed that GTK-transgenic islets and GTK-expressing RINm5F cells exhibit a reduced insulininduced activation of the insulin receptor substrate (IRS-1 and IRS-2)-pathways, partly due to an increased basal activity of these. GTK was found to associate with and phosphorylate the SH2 domain adapter protein SHB, which could explain many of the GTK-dependent effects both in vitro and in vivo. In summary, the present work suggests that the novel tyrosine kinase GTK is involved in various signal transduction pathways, regulating different cellular responses, such as proliferation, differentiation and survival.</p>

<p>Formation of blood vessels occurs through two processes: vasculogenesis and angiogenesis, which are regulated by various growth factors such as vascular endothelial growth factor, fibroblast growth factor and platelet-derived growth factor. The present study was carried out in order to investigate the roles of the Shb and Cbl proteins in growth factor-mediated signal transduction and blood vessel formation. Shb was found to be involved in NGF-stimulated Rap1 signaling in PC12 cells by forming a complex with CrkII and a 130-135 kDa protein. The Rap1 signaling pathway contributed to NGF-dependent neurite outgrowth. In immortomouse brain endothelial (IBE) cells, Shb increased cell spreading, migration and cytoskeletal rearrangements. Such effects may partly be due to altered Rap1 activation in Shb overexpressing IBE cells. Shb was required for tubular morphogenesis in collagen gels in the presence of FGF-2. In embryoid bodies (EBs) derived from murine embryonic stem cells, Shb up-regulated both VEGFR2 and Tal1 expression at early stages of EB development and thus promoted blood vessel formation both in the absence and in the presence of growth factors. In IBE cells, Cbl positively regulated FGF-2 signaling and increased cell proliferation. Mutation of RING finger alone did not affect blood vessel formation in EBs. However, EBs overexpressing the oncogenic form Cbl 70Z, which had a deletion of the linker region and the first cysteine of the RING finger, exhibited intense CD31 positive sheet-like staining and blood vessel. The results suggested that Cbl had dual roles in endothelial cells: it promoted FGF-2-induced proliferation whereas down-regulated proliferation of endothelial progenitor cells.</p><p>The present work suggests that Shb and Cbl play a crucial role in cell differentiation and blood vessel formation.</p>

Protein tyrosine kinases play an important role in the regulation of various cellular processes such as growth, differentiation and survival. GTK, a novel SRC-like cytoplasmic tyrosine kinase, was recently cloned from a mouse insulinoma cell line and the present work was conducted in order to find a biological function of GTK in insulin producing and neuronal cells. It was observed that kinase active GTK-mutants, expressed in RINm5F cells, transferred to the cell nucleus and increased the levels of the cell cycle regulatory protein p27KIP1, reduced cell growth and stimulated glucagon mRNA expression. Furthermore, wild type GTK induces neurite outgrowth in the rat adrenal pheochromocytoma PC12 cell line, through activation of the RAP1-pathway, suggesting a role of GTK for cell differentiation. Studies using transgenic mice, expressing GTK under the control of the rat insulin 1 promoter, demonstrated a dual role of GTK for β-cell growth: Whereas GTK increases the β-cell mass and causes enhanced β-cell proliferation in response to partial pancreatectomy it also induced β-cell death in response to proinflammatory cytokines and impaired the glucose tolerance in mice treated with the β-cell toxin streptozotocin suggesting a possible role of GTK for β-cell destruction in Type 1 diabetes. We have also observed that GTK-transgenic islets and GTK-expressing RINm5F cells exhibit a reduced insulininduced activation of the insulin receptor substrate (IRS-1 and IRS-2)-pathways, partly due to an increased basal activity of these. GTK was found to associate with and phosphorylate the SH2 domain adapter protein SHB, which could explain many of the GTK-dependent effects both in vitro and in vivo. In summary, the present work suggests that the novel tyrosine kinase GTK is involved in various signal transduction pathways, regulating different cellular responses, such as proliferation, differentiation and survival.

Formation of blood vessels occurs through two processes: vasculogenesis and angiogenesis, which are regulated by various growth factors such as vascular endothelial growth factor, fibroblast growth factor and platelet-derived growth factor. The present study was carried out in order to investigate the roles of the Shb and Cbl proteins in growth factor-mediated signal transduction and blood vessel formation. Shb was found to be involved in NGF-stimulated Rap1 signaling in PC12 cells by forming a complex with CrkII and a 130-135 kDa protein. The Rap1 signaling pathway contributed to NGF-dependent neurite outgrowth. In immortomouse brain endothelial (IBE) cells, Shb increased cell spreading, migration and cytoskeletal rearrangements. Such effects may partly be due to altered Rap1 activation in Shb overexpressing IBE cells. Shb was required for tubular morphogenesis in collagen gels in the presence of FGF-2. In embryoid bodies (EBs) derived from murine embryonic stem cells, Shb up-regulated both VEGFR2 and Tal1 expression at early stages of EB development and thus promoted blood vessel formation both in the absence and in the presence of growth factors. In IBE cells, Cbl positively regulated FGF-2 signaling and increased cell proliferation. Mutation of RING finger alone did not affect blood vessel formation in EBs. However, EBs overexpressing the oncogenic form Cbl 70Z, which had a deletion of the linker region and the first cysteine of the RING finger, exhibited intense CD31 positive sheet-like staining and blood vessel. The results suggested that Cbl had dual roles in endothelial cells: it promoted FGF-2-induced proliferation whereas down-regulated proliferation of endothelial progenitor cells. The present work suggests that Shb and Cbl play a crucial role in cell differentiation and blood vessel formation.

<p>Stimulation of the T cell receptor (TCR) induces tyrosine phosphorylation of numerous intracellular proteins, leading to activation of the interleukin-2 (IL-2) gene in T lymphocytes. Shb is a ubiquitously expressed adapter protein, with the ability to associate with the T cell receptor and several signaling proteins in T cells, including: the TCR ζ-chain, LAT, PLC-γ1, Vav, SLP-76 and Gads. Jurkat T cells expressing Shb with a mutation in the SH2 domain, exhibited reduced phosphorylation of several proteins and abolished activation of the MAP kinases ERK1, ERK2 and JNK, upon CD3 stimulation. The TCR induced Ca²⁺ response in these cells was abolished, together with the activation of the IL-2 promoter via the transcription factor NFAT. Consequently, IL-2 production was also perturbed in these cells, compared to normal Jurkat T cells. Shb was also seen to associate with the β and γ chains of the IL-2 receptor, upon IL-2 stimulation, in T and NK cells. This association occurred between the Shb SH2 domain and Tyr-510 of the IL-2R β chain. The proline-rich domains of Shb were found to associate with the tyrosine kinases JAK1 and JAK3, which are important for STAT-mediated proliferation of T and NK cells upon IL-2 stimulation. Shb was also found to be involved in IL-2 mediated regulation of apoptosis. These findings indicate a dual role for Shb in T cells, where Shb is involved in both T cell receptor and IL-2 receptor signaling. </p><p>A Shb homologue, Shf was identified, and seen to associate with the PDGF-α-receptor. Shf shares high sequence homology with Shb and a Shd (also of the Shb family) in the SH2 domain and in four motifs containing putative tyrosine phosphorylation sites. When Shf was overexpressed in fibroblasts, these cells displayed significantly lower rates of apoptosis than control cells in the presence of PDGF-AA. These findings suggest a role for the novel adapter Shf in PDGF-receptor signaling and regulation of apoptosis.</p>