The epidemiology of urinary tract infection (UTI) in the population of Vännäs (8 000 inhabitants) was studied during one year. The annual incidence increased from 0.5% in the first decade of life to more than 10% in the age group 90-100 years. Male UTI comprised only 13% of the episodes, increased after middle age and contributed 4 0% by &gt; 80 years of age. At 17 PHC centres (PHCCs) a prevalence study (McPHC) of mainly uncomplicated UTI was performed. Most episodes were acutely symptomatic (lower 75%, upper 5%). Microscopy of wet-stained urinary sediment with a minimum of moderate amount of bacteria and/or 5 leukocytes per high power field (4 00 x) as breakpoint resulted in a desired high sensitivity (97%) and 86% efficacy in acutely symptomatic patients. Diagnosis of bacteriuria using Uricult dipslides yielded acceptable results with an overall efficacy of 88%. Nitrite test and Uriglox showed an unacceptable low mean sensitivity ofR56 and 69%, respectively. A positive nitrite, sediment or Uricult , when used in combination, was optimal in diagnosing UTI with a sensitivity of 98% in acutely symptomatic patients during their office visits. The average risk of drug resistance was 17% in the Vännäs study. Sensicult satisfactorily predicted drug sensitivity (93%) but not bacterial drug resistance (50%). Using Uricult with classification of bacteriuria by Gram-grouping, lactose and catalase reactions for targeting UTI therapy, according to local guidelines, resulted in a similar low risk (6 %) of prescribing drugs to which the organisms were resistagt as when using Sensicult (7%). This development of the Uricult method is simple and can be recommended for office practice in PHC. The spectrum of bacteria causing UTI and their drug resistance was more associated with the selection of patients, sex and age than with symptoms. The pattern of drug resistance was little influenced by UTI history and the mean pretherapy resistance for the seven drugs tested in McPHC was low (7%). Drug resistance was increased in failure (mean 24%) but not in early or repeated recurrence. In McPHC therapy resulted in 8% bacteriological failure and 12% early recurrence, irrespective of whether the bacteria were classified as sensitive or resistant in vitro to the drug given. Thus, in order to be of prognostic value for therapy of uncomplicated UTI, high-level breakpoints focusing more on peak urinary drug concentrations need to be studied. UTI symptoms in McPHC were eradicated in only 2/3 of the bacterio- logically cured episodes and in 1/3 of the failures at control 1-3 days posttherapy showing that symptoms are an unreliable indicator of UTI. From current literature, it seems unlikely that asymptomatic bacteriuria (ABU) plays a major role in the development of uremia due to chronic pyelonephritis. With the exception of ABU in pregnancy, therapy seems to yield no benefit. Omitting posttherapy bacteriuria controls in patients with symptoms eradicated, at least in women with uncomplicated UTI, would lead to considerable savings both for patients and the health care system. / <p>Diss. (sammanfattning) Umeå : Umeå universitet, 1988, härtill 6 uppsatser.</p> / digitalisering@umu

Statins, common cholesterol control drugs, are appreciated to have promising anticancer activity through inhibition of the mevalonate pathway. Several lines of evidence suggest that certain tumours are susceptible to statins, but the underlying molecular features arbitrating this sensitivity remain unknown. We hypothesize that (i) not all statins will behave equivalently in the context of anticancer therapy, and (ii) a molecularly-defined subset of tumours are intrinsically sensitive to statins. My objectives have therefore been to further our understanding of functional differences between statins influencing their anticancer effects, and to investigate molecular features associated with statin sensitivity in breast cancer. Specifically, this thesis addresses two aims: (i) to characterize differential interactions between four statins and the xenobiotic transporter P-glycoprotein (P-gp; also known as ABCB1), and (ii) to identify molecular features associated with fluvastatin and lovastatin sensitivity in breast tumour cell lines. We first characterized the interactions of statins with P-gp in vitro and in multidrug-resistant (MDR) tumour cells. While lovastatin could directly bind to P-gp and modulate MDR, no significant interactions were observed with fluvastatin. Fluvastatin may therefore be appropriate for use in unselected patients, to avoid adverse drug interactions with coadministered P-gp substrate chemotherapeutics. Fluvastatin has also shown promise in breast cancer treatment, where molecular features predictive of statin sensitivity would be particularly valuable. A panel of 19 immortalized breast cell lines was therefore characterized for sensitivity to fluvastatin and lovastatin. Relatively statin-sensitive cells underwent apoptosis upon statin treatment, and were more likely to have an estrogen receptor alpha (ERα)-negative, basal-like phenotype. By mining available baseline gene expression data, a candidate 10-gene signature predictive of fluvastatin sensitivity was also generated. Taken together, this research provides insight into molecular markers of statin sensitivity that may facilitate fast-tracking of these drugs to clinical trials in subsets of cancer patients most likely to respond.

Fluctuations, or "noise", can play a key role in determining the behaviour of living systems. The molecular-level fluctuations that occur in genetic networks are of particular importance. Here, noisy gene expression can result in genetically identical cells displaying significant variation in phenotype, even in identical environments. This variation can act as a basis for natural selection and provide a fitness benefit to cell populations under stress. This thesis focuses on the development of new conceptual knowledge about how gene expression noise and gene network topology influence drug resistance, as well as new simulation techniques to better understand cell population dynamics. Network topology may at first seem disconnected from expression noise, but genes in a network regulate each other through their expression products. The topology of a genetic network can thus amplify or attenuate noisy inputs from the environment and influence the expression characteristics of genes serving as outputs to the network. The main body of the thesis consists of five chapters: 1. A published review article on the physical basis of cellular individuality. 2. A published article presenting a novel method for simulating the dynamics of cell populations. 3. A chapter on modeling and simulating replicative aging and competition using an object-oriented framework. 4. A published research article establishing that noise in gene expression can facilitate adaptation and drug resistance independent of mutation. 5. An article submitted for publication demonstrating that gene network topology can affect the development of drug resistance. These chapters are preceded by a comprehensive introduction that covers essential concepts and theories relevant to the work presented.

Purpose: Control of micrometastatic ovarian cancer in the peritoneal cavity remains a major objective in post-surgical treatment. The purpose of this project was to investigate the efficacy and toxicity of targeted alpha therapy (TAT) for ovarian cancer in vitro and in vivo in animal models and to select the optimal targeting vector for an ovarian cancer clinical trial. Animal models of ovarian, breast and prostate cancer were developed and for further TAT; a phase I melanoma clinical trial was supported, paving the way for an ovarian cancer clinical trial. Methods: The expression of the turnor-associated antigens (Her2, MUC1, uPAfuPAR) on cancer cell line, animal model xenografts and human ovarian cancer tissue was tested by immunostaining. MTS and TUNEL assays were used to evaluate cell killing of alpha conjugates in monolayer and spheroids. Toxicity and maximum tolerance doses for different vectors were tested and determined in vivo. Pharmacokinetics was studied for different time points and different parameters. The antiproliferative effect of 213Bi-C595 and 213Bi-PAI2 was tested at 9 days post-peritoneal cell inoculation of the ovarian cancer cell line OVCAR3. The treatment efficacy of 213Bi-Herceptin was tested at a 2 days post-subcutaneous breast cancer cell BT474 inoculation. Mice were injected (i.p) with various concentrations of alpha conjugates (AC). Changes in cancer progression were assessed by girth size and tumor size. Results: uPA/uPAR and MUCI are expressed on ovarian cancer cell lines and more than 45% ovarian cancer tissue, while HER2 was only positive in one cell line and was positive in less than 15% of ovarian cancer tissues. The ACs can target and kill cancer cells in vitro in a dose dependent fashion. TUNEL positive cells were found after incubation with the different ACs. PAI2 and C595 vectors were selected for in vivo ascites model study of OVCARJ cell with high expression. Delayed and acute toxicity in animal models showed that radiation nephropathy was the cause of body weight loss. Biodistribution studies showed that kidney was the major uptake organ. L-lysine can reduce kidney uptake for 213Bi-PAI2, but no significant differences were found. A single ip injection of 213Bi-C595 or 213Bi-PAI2 can inhibit ascites growth, whereas, 213Bi-Herceptin can inhibit breast cancer growth in a nude mice model. Conclusion: 213Bi labelled targeting vectors can specifically target ovarian cancer cells in vitro and inhibit tumor growth in vivo. These ACs may be useful agents for the treatment of ovarian cancer at the minimum residual disease stage.

ABSTRACT Renal cell carcinoma (RCC), the most common type of kidney cancer, is a highly metastatic disease. Late stage metastatic RCC is essentially incurable and lethal. The average survival of patients, following metastatic RCC, is about 4 months and only 10% of patients survive for one year. None of the currently available chemotherapy, radiotherapy, hormonal or biological therapies have a significant impact on the progression of the disease. Novel chemotherapeutics are urgently required for the treatment of this deadly disease. The mechanisms that pose the greatest challenges to chemotherapeutics are resistance of tumour cells to apoptosis, tumour angiogenesis and multi-drug resistance. Resistance to apoptosis may be mediated by the up-regulation of anti-apoptotic proteins, especially Bcl-2 and Bcl-XL, and/or by the down- regulation of pro-apoptotic proteins, particularly Bax. Angiogenesis is pivotal for tumour growth and metastasis. Of all identified pro-angiogenic molecules, vascular endothelial growth factor (VEGF) is considered to be a key molecule. Drug resistance is thought to be mediated by the up-regulation of multi-drug resistance molecules such as MDR-1 and MRP-1. Up-regulation of Bcl-2 also confers drug resistance to cancer cells. The main hypothesis of this thesis was that treatment targets of metastatic RCCs are likely to multifactorial and that inhibition of molecules that regulate the processes of apoptosis, angiogenesis and multidrug resistance are likely to be better targets than those that regulate only one of these processes. In this regard, the transcription factor nuclear factor kappaB (NF-kB) meets the criterion, regulating the apoptotic, angiogenic and multi-drug resistance pathways of cancer cells. Its inhibition appeared to be an attractive strategy for the treatment of metastatic RCC. Many studies have demonstrated an association between the over-expression NF-kB and RCC. Thus, the major aim of this thesis was to explore the anti-cancer effect of pyrrolidine dithiocarbamate (PDTC), a potent NF-kB inhibitor on human metastatic RCC cell lines. The thesis is divided into seven Chapters. In Chapter 1, the literature on RCC, NF-B and the role of NF-kB in RCC development and progression are reviewed. The rationale for the inhibition of NF-kB as a potential anti-RCC strategy using PDTC is established. During the course of this research, the use of PDTC as an anti-cancer agent has risen to prominence. Chapter 2 describes the materials and methods used in the project. In Chapter 3, the expression of NF-kB in human kidney and the RCC cell lines, ACHN and SN12K1, was established. The proof of hypothesis that NF-kB inhibition using PDTC is an effective anti-cancer strategy was demonstrated. PDTC was selectively toxic to the RCC cell lines, but not to normal human kidney cells. PDTC induced apoptosis and inhibited proliferation of the RCC cells. PDTC also inhibited NF-kB, its upstream regulatory molecules such as the inhibitory protein family of the IkBs, and the kinase IKK complex. PDTC also inhibited anti-apoptotic Bcl-2 and Bcl-XL, but not pro-apoptotic Bax. Chapter 4 demonstrated the in vitro and ex vivo anti-angiogenic and anti-metastatic effects of PDTC. Protein microarrays for angiogenic factors produced controversial results. PDTC inhibited epidermal growth factor (EGF) produced in endothelial cells. VEGF had neutral effect on angiogenesis under the experimental conditions used. In the RCC cell lines, several pro-angiogenic molecules were modulated. Interestingly, the pro-angiogenic molecule interleukin (IL)-8 was up-regulated in both RCC cell lines. The monocyte chemoattractant protein-1 (MCP-1) was decreased in ACHN cells, but increased in SN12K1 cells. The implications of these controversial findings are discussed. Chapter 5 demonstrated the ability of PDTC to overcome drug resistance in a synergism with cisplatin. Individual non-toxic concentrations of PDTC and cisplatin, when combined, induced significant toxicity of RCC cell lines. The synergistic effect was not mediated by the inhibition of NF-kB, but rather through the inhibition of transcriptional activation of NF-kB. Bcl-2 rather than MDR-1 or the regulatory protein MRP-1 may be important in overcoming drug resistance in RCC. Chapter 6 showed the anti-cancer effect of PDTC in an animal model of RCC. PDTC significantly decreased the growth of RCC implanted in the kidney of severe combined immunodeficiency (SCID) mice. PDTC inhibited NF-kB and was not toxic to normal cells. The expression of Bcl-2, Bcl-XL and Bax were contradictory to the in vitro findings and a theory about the spread of RCC based on these findings is discussed. In Chapter 7, the findings are summarised. A case for PDTC as a potential therapeutic agent for RCC is established. Under the experimental conditions used, PDTC was demonstrated to be an effective anti-RCC agent by targeting the three most important characteristics of RCC that pose the greatest challenges to chemotherapeutics: resistance of tumour cells to apoptosis, tumour angiogenesis and multi-drug resistance. PDTC was selectively toxic to RCC, but not to normal renal cells. Thus PDTC appears to be a promising anti-cancer agent. This is supported by the current increase in interest, and in the number of publications, on the use of PDTC in several cancers. Some future directions are also discussed in this Chapter. These include, but are not limited to, an investigation of what is protecting normal cells from the toxicity of PDTC, the creation of an Australian database on RCC, and the characterisation of RCC based on NF-kB expression.

Diss. (sammanfattning) Stockholm : Karol inst., 2003. / Härtill 6 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.