| 1. |
- Bodén, Ida, 1977-
(författare)
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Near infrared and skin impedance spectroscopic in vivo measurements on human skin : development of a diagnostic tool for skin cancer
- 2011
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Every year approximately 2800 Swedes are diagnosed with malignant melanoma, the form of cancer that is most rapidly increasing in incidence in the Western world. The earlier we can identify and diagnose a malignant melanoma, the better is the prognosis. In Sweden, 155 000 benign naevi, harmless skin tumours or moles, are surgically excised each year, many of them because melanoma cannot be dismissed by non-invasive methods. The excisions result in substantial medical costs and cause unrest and suffering of the individual patient. For untrained physicians, it is often difficult to make an accurate diagnosis of melanoma, thus a tool that could help to strengthen the diagnosis of suspected melanomas would be highly valuable. This thesis describes the development and assessment of a non-invasive method for early skin cancer detection. Using near infrared (NIR) and skin impedance spectroscopy, healthy and diseased skin of various subjects was examined to develop a new instrument for detecting malignant melanoma. Due to the complex nature of skin and the numerous variables involved, the spectroscopic data were analysed multivariately using Principal Component Analysis (PCA) and partial leas square discriminant analysis (PLS-DA). The reproducibility of the measurements was determined by calculating Scatter Values (SVs), and the significance of separations between overlapping groups in score plots was determined by calculating intra-model distances. The studies indicate that combining skin impedance and NIR spectroscopy measurements adds value, therefore a new probe-head for simultaneous NIR and skin impedance measurements was introduced. Using both spectroscopic techniques it was possible to separate healthy skin at one body location from healthy skin at another location due to the differences in skin characteristics at various body locations. In addition, statistically significant differences between overlapping groups of both age and gender in score plots were detected. However, the differences in skin characteristics at different body locations had stronger effects on the measurements than both age and gender. Intake of coffee and alcohol prior to measurement did not significantly influence the outcome data. Measurements on dysplastic naevi were significantly separated in a score plot and the influence of diseased skin was stronger than that of body location. This was confirmed in a study where measurements were performed on 12 malignant melanomas, 19 dysplastic naevi and 19 benign naevi. The malignant melanomas were significantly separated from both dysplastic naevi and benign naevi. Overall, the presented findings show that the instrument we have developed provides fast, reproducible measurements, capable of distinguishing malignant melanoma from dysplastic naevi and benign naevi non-invasively with 83% sensitivity and 95% specificity. Thus, the results are highly promising and the instrument appears to have high potential diagnostic utility.
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| 2. |
- Henriksson, Otto, 1976-
(författare)
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Protection against cold in prehospital trauma care
- 2012
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: Protection against cold is vitally important in prehospital trauma care to reduce heat loss and prevent body core cooling. Objectives: Evaluate the effect on cold stress and thermoregulation in volunteer subjects byutilising additional insulation on a spineboard (I). Determine thermal insulation properties of blankets and rescue bags in different wind conditions (II). Establish the utility of wet clothing removal or the addition of a vapour barrier by determining the effect on heat loss within different levels of insulation in cold and warm ambient temperatures (III) and evaluating the effect on cold stress and thermoregulation in volunteer subjects (IV). Methods: Aural canal temperature, sensation of shivering and cold discomfort was evaluated in volunteer subjects, immobilised on non-insulated (n=10) or insulated (n=9) spineboards in cold outdoor conditions (I). A thermal manikin was setup inside a climatic chamber and total resultant thermal insulation for the selected ensembles was determined in low, moderate and high wind conditions (II). Dry and wet heat loss and the effect of wet clothing removal or the addition of a vapour barrier was determined with the thermal manikin dressed in either dry, wet or no clothing; with or without a vapour barrier; and with three different levels of insulation in warm and cold ambient conditions (III). The effect on metabolic rate, oesophageal temperature, skin temperature, body heat storage, heart rate, and cold discomfort by wet clothing removal or the addition of a vapour barrier was evaluated in volunteer subjects (n=8), wearing wet clothing in a cold climatic chamber during four different insulation protocols in a cross-over design (IV). Results: Additional insulation on a spine board rendered a significant reduction of estimated shivering but there was no significant difference in aural canal temperature or cold discomfort (I). In low wind conditions, thermal insulation correlated to thickness of the insulation ensemble. In greater air velocities, thermal insulation was better preserved for ensembles that were windproof and resistant to the compressive effect of the wind (II). Wet clothing removal or the use of a vapour barrier reduced total heat loss by about one fourth in the cold environment and about one third in the warm environment (III). In cold stressed wet subjects, with limited insulation applied, wet clothing removal or the addition of a vapour barrier significantly reduced metabolic rate, increased skin rewarming rate, and improved total body heat storage but there was no significant difference in heart rate or oesophageal temperature cooling rate (IV). Similar effects on heat loss and cold stress was also achieved by increasing the insulation. Cold discomfort was significantly reduced with the addition of a vapour barrier and with an increased insulation but not with wet clothing removal. Conclusions: Additional insulation on a spine board might aid in reducing cold stress inprolonged transportations in a cold environment. In extended on scene durations, the use of a windproof and compression resistant outer cover is crucial to maintain adequate thermal insulation. In a sustained cold environment in which sufficient insulation is not available, wet clothing removal or the use of a vapour barrier might be considerably important reducing heat loss and relieving cold stress.
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| 3. |
- Lundgren, Peter, 1977-
(författare)
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Protection and treatment of hypothermia in prehospital trauma care : with emphasis on active warming
- 2012
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: In prehospital trauma care active warming is recommended to aid in protection from further cooling. However, scientific evidence of the effectiveness of active warming in a clinical setting is scarce. Also, evaluating the effectiveness of active warming, especially in harsh ambient conditions, by objective measures, is difficult. Objective: To evaluate the effectiveness of field applicabe heat sources (I) and to evaluate active warming intervention in a prehospital clinical setting (II and III). To evaluate reliability and validity of the Cold Discomfort Scale (CDS), a subjective judgement scale for assessment of the thermal state of patients in a cold environment (IV). Methods: In a laboratory trial, non-shivering hypothermic subjects (n=5), were cooled in 8 ºC water followed by spontaneous warming, a charcoal heater, two flexible hot-water bags or two chemical heat pads, all applied to the chest and upper back (I). Oesophageal temperature, skin temperature, heat flux, oxygen consumption, respiratory rate and, heart rate were measured. In two clinical randomized trials, shivering patients during road and air ambulance transport (II) and during field treatment (III) were randomized to either passive warming alone (n=22 and n=9) or to passive warming with the addition of a chemical heat pad (n=26 and n=11). Body core temperature, respiratory rate, heart rate, blood pressure (II) and the patients’ subjective sensation of thermal comfort (II and III) were measured. In a laboratory trial, shivering subjects were exposed to – 20 ºC (n=22). The CDS was evaluated regarding reliability, defined as test-retest stability, and criterion validity, defined as the ability to detect changes in cold discomfort due to changes in cumulative cold stress (IV). Results: In non-shivering hypothermic subjects postcooling afterdrop was significantly less for the chemical heat pads, but not for the hot water bags and the charcoal heater, compared to spontaneous warming (I). Temperature drop during the entire warming phase was significantly less for all the heat sources respectively, compared to spontaneous warming (I). During road and air ambulance transport, ear canal temperature was significantly increased and cold discomfort significantly decreased, both in patients assigned to passive warming only, and in patients assigned to additional active warming (II). During field treatment, cold discomfort was significantly reduced in patients assigned to additional active warming, but remained the same in patients assigned to passive warming only (III). Weighted kappa coefficient, describing test-retest stability, was 0.84 (IV). CDS ratings were significantly increased during each 30 minutes interval (IV). Conclusion: In non-shivering hypothermic subjects, heat sources were effective to attenuate afterdrop, when providing high heat content over a large surface area and effective to continue to increase body core temperature when providing sustained high heat content. In shivering trauma patients, adequate passive warming were sufficient treatment to prevent afterdrop, to slowly increase body core temperature, and to reduce cold discomfort. If inadequate passive warming, additional active warming was required to reduce cold discomfort. The CDS, a subjective judgement scale for assessment of the thermal state of patients in a cold environment seemed to be reliable regarding test-retest stability and valid regarding ability to detect change in cumulative cold stress.
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| 4. |
- Natarajan, Balasubramanian, 1981-
(författare)
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New modifiers of insulin signalling identified by interaction screens with ASNA-1 in C. elegans
- 2012
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: Insulin is a hormone released by the pancreatic beta cells in response to elevated levels of nutrients in the blood. Insulin triggers the uptake of glucose, fatty acids and amino acids into the liver, adipose tissue and muscles. Genes regulating insulin signalling are thus of vital importance for metabolic homeostasis and for preventing the development of diabetes. This thesis aims to identify new modifiers of insulin signalling, while carrying out functional studies of a homolog to human arsenite translocating ATPase, ASNA1. ASNA1 activates the insulin signalling pathway and promotes insulin secretion in mammalian cell lines and in Caenorhabditis elegans. A second aim is to better understand how ASNA1 and its interactors regulate sensitivity to the chemotherapeutic drug, cisplatin. Results: Regulators of insulin/IGF signalling (IIS) in C. elegans were identified based on the Larval arrest arrest aspect of the asna-1 depletion phenotype. Sixty-five genes were selected by virtue of their predicted interaction with ASNA-1 and screened for asna-1-like larval arrest upon inactivation of the genes . mrps-2, mrps-10, mrpl-43 encoding mitochondrial ribosomal protein subunits, and enpl-1 encoding an ER chaperone, GRP94 homolog were identified as the genes which when inactivated caused larval arrest without any associated feeding defects. IIS was weaker and insulin secretion was defective in these knockdown animals. ENPL-1 and ASNA-1 proteins interacted with one another both ex vivo and in vitro. ASNA-1 protein and mRNA level swere greatly reduced in enpl-1 mutants and enpl-1(-);asna-1(-) double-mutant worms displayed synthetic lethality. Overexpression of the insulins INS-4 and DAF-28 caused partial rescue of the germline phenotype of enpl-1 mutants, indicating that the phenotype of enpl-1 mutants was due at least in part to insufficient insulin levels. Studies of enpl-1 mutants also helped to understand the role of asna-1 in cisplatin sensitivity. The unfolded protein response (UPR) was induced in asna-1 and enpl-1 knockdown animals. enpl-1 mutants displayed higher sensitivity to cisplatin, when compared to asna-1 mutants and this correlated to higher UPR in enpl-1 knockdown animals. Pharmacological induction of the UPR in intrinsically cisplatin resistant wildtype worms also resulted in increased cisplatin sensitivity. This suggests that manipulation of ENPL-1 levels or of the UPR could enhance the anti-tumoral effects of cisplatin based cancer therapy. With a yeast two hybridscreen 27 putative physical interactors of ASNA-1 were identified. Amongst these candidate swas smn-1, which encodes survival of motor neuron protein homolog. RNAi knockdown of smn-1 caused a larval arrest phenotype similar to asna-1 depleted animals and smn-1 positively regulated IIS, like asna-1. Defects in IIS may be at the level of insulin release because neuropeptide secretion was impaired upon smn-1 knockdown. Further in vitro binding studies showed that SMN-1 and ASNA-1 interacted and inactivation of smn-1 in asna-1 mutants resulted in decreased viability. This implies that SMN-1 is another modifier of ASNA-1 and also a new component in IIS. Conclusion: With a directed RNAi screen and a yeast two hybrid screen several interactors of ASNA-1 that are also IIS modifiers were identified. ENPL-1 and SMN-1 are both involved in insulin release. We also found that induction of the UPR in enpl-1 and asna-1 mutants is a possible mechanism for increased sensitivity to cisplatin.
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| 5. |
- Nyström, Hanna, 1980-
(författare)
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Stromal collagens in colorectal cancer and in colorectal liver metastases : tumour biological implications and a source for novel tumour markers
- 2013
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: Colorectal cancer (CRC) remains one of the leading causes of cancer-related mortality. About 50 % of patients with CRC will develop subsequent liver metastases (CLM). The survival for untreated CLM is only a few months and liver resection provides the only chance for a lasting cure. It is therefore essential to detect CLM early, enabling successful surgical resection and achieving a long-term cure. There are no optimal tumour markers for CRC or CLM. The best marker available is Carcinoembryonic Antigen (CEA), a marker found elevated in about 50-60% of patients with CLM, but also in many other conditions. The main focus of cancer research has been on the malignant cancer cell. However, a tumour consists of more than cancer cells. A major part of all solid tumours is made up by the stroma. The tumour stroma is defined as the non-malignant cells of a tumour such as fibroblasts, the cells of the vascular and immune systems as well as the extracellular matrix (ECM). The basement membrane (BM) is a specialized form of the ECM in which type IV collagen is the major protein component. All epithelial cells need a contact to the BM and the definition of an invasive cancer is the degradation of the BM and the spread of cancer cells beyond this structure. Different metastatic growth patterns of CLM have previously been described, namely the desmoplastic, pushing and replacement type of CLM. These differ in their stromal reaction in the border, which separates the tumour from the normal liver. In this thesis the tumour stroma of CRC and CLM is studied with a special emphasis on stromal collagens. The aim is to investigate whether stromal collagens/ circulating type IV collagen can be used as tumour markers for CRC and CLM, and to compare this to the conventional marker CEA. The circulating type IV collagen level is also measured in liver metastases from other primary tumours than CRC. Furthermore, the differences between the stroma of a primary CRC that metastasizes to the liver when compared to a CRC that never spreads are analysed. Additionally, the metastatic growth pattern of CLM is studied in relation to the primary tumour, stromal components and survival. We also sought out to find whether CRC cell lines possess the trait to produce ECM proteins endogenously, and in response to a normal liver stroma in a novel organotypic model for CLM.Methods: Expression patterns of type I, III and IV collagen were studied by immunofluorescence (IF), chemical staining and immunohistochemistry (IHC) in normal colorectal tissue, normal liver, CRC, CLM, benign liver lesions and in liver metastases of other origin than CRC. Circulating plasma levels of type IV collagen were analysed in healthy controls, patients with CRC (T stage I-III) and in patients with CLM. Samples were analysed at the time of diagnosis, during and after oncological and surgical treatment and at the time of relapsing or progressive disease. Additionally, circulating levels were analysed in patients with benign liver lesions and in liver metastases of other origin than CRC. The metastatic growth pattern of CLM was classified according to earlier descriptions. CRC cell lines were studied regarding their production of type IV collagen. The growth, invasiveness and stromal production in CRC cell lines were also investigated in a new organotypic model for CLM using human liver specimens.Results: Circulating type IV collagen levels are increased in patients with CLM and other epithelial-derived liver metastases, and is found normal in patients with primary CRC (stage I-III), with liver metastases from tumours of non-epithelial origin, benign liver lesions and in healthy controls. The type IV collagen levels in patients with CLM reflect the tumour burden in the liver, decreases in response to therapy and is found increased in progressive or relapsing disease. The combination of circulating type IV collagen and CEA increased the sensitivity and specificity for detecting CLM. Livermetastatic CRC displayed an increased stromal production when compared to non-metastatic CRC, with an increased type IV collagen expression in the direct vicinity of the CRC cells. The earlier described growth patterns of CLM were verified, with the pushing type of CLM associated with a short survival and poor outcome. Furthermore, CRC cell lines possess the trait of endogenously producing type IV collagen. The novel organotypic liver model revealed that CRC cell lines grown in the context of normal liver stroma, devoid of other cells, does not elicit a desmoplastic reaction.Conclusion: Circulating type IV collagen is a promising tumour marker for CLM, where the levels reflect the hepatic tumour burden and can detect disease relapse after liver surgery. The combination of the tumour markers CEA and type IV collagen is superior to CEA alone. The stromal composition of primary CRC predicts the risk of subsequent CLM and the metastatic growth pattern of CLM is related to survival.
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| 6. |
- Öhlund, Daniel, 1979-
(författare)
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Basement membrane collagens in pancreatic cancer : novel stroma-derived tumor markers and regulators of cancer cell growth
- 2010
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: Among the common malignancies, pancreatic cancer has the shortest long-term survival. The aggressive, rapid, and infiltrative growth pattern of pancreatic cancer, together with the lack of specific symptoms, often leads to late diagnosis. Metastases are frequently found at the time of diagnosis, which prevents curative surgical treatment. Good tumor markers would enable early detection, thus improving the prognosis. Unfortunately, no such markers are available in the clinic. The tumor stroma is defined as the non-malignant cells and the extracellular matrix (ECM) of a cancer. Pancreatic cancer is characterized by an abundant tumor stroma, rich in ECM proteins such as collagens, which have been shown to play important roles in tumor progression. Furthermore, pancreatic cancer cells produce large quantities of ECM proteins, especially the basement membrane (BM) protein type IV collagen. All epithelial cells are anchored to a BM, which must be degraded in order for an in situ cancer to become invasive. Matrix metalloproteinases (MMPs) are enzymes involved in BM degradation. In this thesis, the tumor stroma of pancreatic cancer is studied, focusing on the BM proteins type IV and type XVIII collagen, with the aim to clarify if the stroma could be a source of novel tumor markers for this form of cancer. Additionally, the role of type IV collagen produced by the cancer cells is studied. Methods: Expression patterns of type IV and type XVIII collagen, MMPs involved in collagen degradation, and collagen receptors (integrins) were studied by immunoflourescence in both normal and pancreatic cancer tissue, and in pancreatic cancer cell lines. Circulating plasma levels of type IV and type XVIII collagen and conventional tumor markers (TPS, Ca 19-9, CEA and Ca 125) were measured in controls and pancreatic cancer patients at the time of diagnosis and after treatment. The role of cancer cell produced type IV collagen was studied in human pancreatic cancer cell lines by functional blocking of integrin receptors (integrin a1, a2 and b1) and integrin-binding sites on type IV collagen, and by siRNA-induced down-regulation of type IV collagen synthesis. Proliferation was analyzed by a luminescence based cell viability assay, migration by time-lapse microscopy, and apoptosis by M30-neoepitope detection. Results: MMPs involved in BM degradation were upregulated in pancreatic cancer tissue. The expression of type XVIII collagen shifted from a general BM expression pattern in normal tissue, to mainly being found in the tumor vasculature in pancreatic cancer. Type IV collagen, on the other hand, remained highly expressed in the vicinity of the cancer cells. The a1, a2, and b1 integrin receptors were highly expressed at the cancer cell surface. Both down-regulation of type IV collagen synthesis and blocking the integrin/type IV collagen interaction decreased cell proliferation and migration. The proliferative capacity was rescued by the addition of exogenous type IV collagen. Furthermore, the circulating levels of both type IV and type XVIII collagen were increased in pancreatic cancer patients at the time of diagnosis compared to controls. After treatment, the levels were normalized for type XVIII collagen, whereas the levels of type IV collagen remained high after surgery. High postoperative levels of type IV collagen were associated with short overall survival. A similar association to short survival was found for preoperative type XVIII collagen levels. No such associations to survival could be detected for the conventional markers. Conclusion: The results of this thesis show that type IV and type XVIII collagens can serve as tumor markers for pancreatic cancer with advantages compared to conventionally used markers. Additionally, evidence is provided of an autocrine loop, involving type IV collagen and its integrin receptors, with importance for retaining a proliferative and migratory phenotype in pancreatic cancer cells.
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