Den enkla naturen och andligheten : en fallstudie om vad den fysiska miljön kan betyda på en pilgrimsvandring

This paper treats the physical environments influence on the spritual experience that comes with pilgrimage. The paper deals with a case study on a two-day pilgrimage in Sweden. The method triangulating has been used in the forms; participating observation, interviews and one poll. The question at issue has also included what spiritualism really is för the persons in the paper. The interviews are also concerned with the general environmental significance on pilgrimages. The study shows that the wide open landscape and the fact that you can see so far ahead has helped the spiritual experience most. The persons are also affected generally of details in the environment as water, plants and animals. The fact that there were churches on the way were very important to the spiritual experience and the length of the road was also of great importance. The spiritual experiences environmental influence also has to do with how the pepole define spiritualism. To sum everything up; the simple environment and the context that the Pilgrimagecenter has for its walkings has affected the participants spiritual experience on this walking (pilgrimage).Denna uppsats behandlar den fysiska miljöns betydelse för den andliga upplevelsen vid pilgrimsvandring. Uppsatsen består av en fallstudie på en tvådagars pilgrimsvandring i Sverige. Metoden triangulering har använts i form av deltagande observation, intervjuer och en enkät. De tillfrågade har beskrivit vad andlighet är för dem. Intervjuerna har även gällt miljöns betydelse generellt på pilgrimsvandring. Studien visar att det vida öppna landskapet och att man ser så långt har hjälpt den andliga upplevelsen mest. Personerna påverkas också generellt av detaljer i miljön såsom vatten, växter och djur. Att det finns kyrkor på vägen var mycket viktigt för den andliga upplevelsen och vägens längd hade stor betydelse. Miljöns påverkan på den andliga upplevelsen beror också på hur personerna definierar andlighet. Sammantaget har den enkla miljön och det sammanhang som Pilgrimscentrum har för sina vandringar påverkat deltagarnas andliga upplevelse på den här vandringen

Studies on o-glycosylation of mucin-type proteins and their binding to antibodies, bacterial toxins and viral receptors

Carbohydrates are ubiquitous on the surface of all cells in mammals where they are involved in interactions with the surroundings (extracellular matrix), other cells (including self and non - self) and microbes (bacteria and virus). Carbohydrate - protein inte ractions in nature are often mediated via multivalent binding where the combined strength of multiple receptor - ligand interactions results in a binding that is highly specific and strong. In this thesis we have produced proteins that are densely decorated with carbohydrate determinants in order to study the glycosylation capacity of cell lines (paper I) and generate efficient binders of antibodies (paper II), bacterial toxins (paper III) and virus receptors such as the influenza hemagglutinin (paper IV). P - selectin glycoprotein ligand - 1 (PSGL - 1) is a mucin - type protein that is heavily substituted with O - glycans. PSGL - 1 genetically fused to mouse IgG 2b Fc forms a dimeric PSGL - 1/mIgG 2b mucin - type fusion protein. In paper I, PSGL - 1/mIgG 2b was produced in Sf9 ( S podoptera frugiperda ) and Hi - 5 ( Trichoplusia ni ) cell lines. The mucin - type protein was used as a probe to analyze the O - glycosylation capacity of the se cell lines, which today are used for the commercial production of recombinant proteins and vaccine co mponents. Liquid chromatography - mass spectrometry (LC - MS) revealed that the O - glycosylation was more abundant and complex than previously reported which may limit their use for the production of therapeutic proteins. The glycosylation of PSGL - 1/mIgG 2b may be tailored by producing the protein in genetically engineered cell lines. Rational glycan design is achieved by transfecting cells with plasmids encoding PSGL - 1/mIgG 2b together with specific glycosyltransferases that expand the glycosylation capacity of the cells. In paper II, genetically engineered Chinese Hamster Ovary (CHO) cells were used to produce PSGL - 1/mIgG 2b carrying blood group A and B determinants on type 1, 2 and 3 outer core saccharide chains. The multivalent mucins could adsorb chain type - sp ecific anti - A antibodies, which indicate a prospective use of the mucins in immunoadsorption (IA) columns. IA columns are used to remove anti - A and anti - B reactive antibodies prior to organ transplantation across the blood group ABO barrier. In paper III a nd IV, genetically engineered CHO cells were used to produce high affinity binders of Shiga toxin 1 and 2 (Stx1 and Stx2) and avian influenza hemagglutinin (H5) . Biacore biosensor assays indicated that PSGL - 1/mIgG 2b carrying the blood group P1 determinant in multiple copies bound with high affinity to Stx1 and Stx2, while PSGL - 1/mIgG 2b presenting multiple copies of Sia α 2,3Gal on different O - linked cores bound with high affinity to avian influenza H5. It remains to be shown if PSGL - 1/mIgG 2b can competitively inhibit and sterically block toxin and viral attachment to the cell surface. In conclusion, PSGL - 1/mIgG 2b carrying specific carbohydrates is a versatile tool that can be used in a range of applications where the multivalency confers a biologically relevan t binding

Erythrocytes Prevent Degradation of Carnosine by Human Serum Carnosinase

The naturally occurring dipeptide carnosine (-alanyl-L-histidine) has beneficial effects in different diseases. It is also frequently used as a food supplement to improve exercise performance and because of its anti-aging effects. Nevertheless, after oral ingestion, the dipeptide is not detectable in human serum because of rapid degradation by serum carnosinase. At the same time, intact carnosine is excreted in urine up to five hours after intake. Therefore, an unknown compartment protecting the dipeptide from degradation has long been hypothesized. Considering that erythrocytes may constitute this compartment, we investigated the uptake and intracellular amounts of carnosine in human erythrocytes cultivated in the presence of the dipeptide and human serum using liquid chromatography–mass spectrometry. In addition, we studied carnosine’s effect on ATP production in red blood cells and on their response to oxidative stress. Our experiments revealed uptake of carnosine into erythrocytes and protection from carnosinase degradation. In addition, no negative effect on ATP production or defense against oxidative stress was observed. In conclusion, our results for the first time demonstrate that erythrocytes can take up carnosine, and, most importantly, thereby prevent its degradation by human serum carnosinase

Correlation between β-catenin mutations and expression of Wnt-signaling target genes in hepatocellular carcinoma

Aberrant Wnt-signaling caused by mutants of β-catenin, a key regulator of the canonical Wnt-signaling pathway, is frequently detected in cancer. Only recently, it was suggested that in hepatocellular carcinoma (HCC) the expression of the target gene glutamine synthetase (GS) is a highly reliable marker for the identification of β-catenin mutations. In order to prove this hypothesis, 52 samples from human hepatocellular carcinomas were analysed for the activation of β-catenin and the expression of GS. In total, 45 samples stained positive for cytoplasmic/nuclear β-catenin. A strong correlation between expression of GS and activated β-catenin (100% of nuclear and 84% of cytosolic) was found. However, among 35 GS positive tumors that were analysed for β-catenin mutations no mutations were detected in 25 GS-positive carcinomas although 24 out of the 25 carcinomas exhibited at least abnormal expression of β-catenin. Since the mutational analysis identified 9 different point mutations of the β-catenin gene including the rare mutation H36P and the yet unknown mutation P44A it was asked whether these mutations may differently effect β-catenin target genes. Therefore, expression plasmids for different mutations were constructed and cotransfected with the TOP-flash luciferase reporter and a reporter carrying the GS-5'-enhancer. The experiments confirmed that there are differences between different β-catenin target sequences and different β-catenin mutations. In addition, the failure that the endogenous expression of GS in GS-negative cells was not induced by the transient transfection experiment indicated that the effect of β-catenin on the GS-5'-enhancer is only one aspect of gene activation induced by β-catenin

Modulation of GLO1 expression affects malignant properties of cells

The energy metabolism of most tumor cells relies on aerobic glycolysis (Warburg effect) characterized by an increased glycolytic flux that is accompanied by the increased formation of the cytotoxic metabolite methylglyoxal (MGO). Consequently, the rate of detoxification of this reactive glycolytic byproduct needs to be increased in order to prevent deleterious effects to the cells. This is brought about by an increased expression of glyoxalase 1 (GLO1) that is the rate-limiting enzyme of the MGO-detoxifying glyoxalase system. Here, we overexpressed GLO1 in HEK 293 cells and silenced it in MCF-7 cells using shRNA. Tumor-related properties of wild type and transformed cells were compared and key glycolytic enzyme activities assessed. Furthermore, the cells were subjected to hypoxic conditions to analyze the impact on cell proliferation and enzyme activities. Our results demonstrate that knockdown of GLO1 in the cancer cells significantly reduced tumor-associated properties such as migration and proliferation, whereas no functional alterations where found by overexpression of GLO1 in HEK 293 cells. In contrast, hypoxia caused inhibition of cell growth of all cells except of those overexpressing GLO1. Altogether, we conclude that GLO1 on one hand is crucial to maintaining tumor characteristics of malignant cells, and, on the other hand, supports malignant transformation of cells in a hypoxic environment when overexpressed

Viability of Glioblastoma Cells and Fibroblasts in the Presence of Imidazole-Containing Compounds

The naturally occurring dipeptide carnosine (-alanyl-L-histidine) specifically attenuates tumor growth. Here, we ask whether other small imidazole-containing compounds also affect the viability of tumor cells without affecting non-malignant cells and whether the formation of histamine is involved. Patient-derived fibroblasts and glioblastoma cells were treated with carnosine, L-alanyl-L-histidine (LA-LH), -alanyl-L-alanine, L-histidine, histamine, imidazole, -alanine, and L-alanine. Cell viability was assessed by cell-based assays and microscopy. The intracellular release of L-histidine and formation of histamine was investigated by high-performance liquid chromatography coupled to mass spectrometry. Carnosine and LA-LH inhibited tumor cell growth with minor effects on fibroblasts, and L-histidine, histamine, and imidazole affected viability in both cell types. Compounds without the imidazole moiety did not diminish viability. In the presence of LA-LH but not in the presence of carnosine, a significant rise in intracellular amounts of histidine was detected in all cells. The formation of histamine was not detectable in the presence of carnosine, LA-LH, or histidine. In conclusion, the imidazole moiety of carnosine contributes to its anti-neoplastic effect, which is also seen in the presence of histidine and LA-LH. Despite the fact that histamine has a strong effect on cell viability, the formation of histamine is not responsible for the effects on the cell viability of carnosine, LA-LH, and histidine

Analysis of cellular and molecular antitumor effects upon inhibition of SATB1 in glioblastoma cells

Background: The Special AT-rich Sequence Binding Protein 1 (SATB1) regulates the expression of many genes by acting as a global chromatin organizer. While in many tumor entities SATB1 overexpression has been observed and connected to pro-tumorigenic processes, somewhat contradictory evidence exists in brain tumors with regard to SATB1 overexpression in glioblastoma and its association with poorer prognosis and tumor progression. On the functional side, initial data indicate that SATB1 may be involved in several tumor cell-relevant processes. Methods: For the detailed analysis of the functional relevance and possible therapeutic potential of SATB1 inhibition, we employ transient siRNA-mediated knockdown and comprehensively analyze the cellular and molecular role of SATB1 in glioblastoma. Results: In various cell lines with different SATB1 expression levels, a SATB1 gene dose-dependent inhibition of anchorage-dependent and –independent proliferation is observed. This is due to cell cycle-inhibitory and pro-apoptotic effects of SATB1 knockdown. Molecular analyses reveal SATB1 knockdown effects on multiple important (proto-) oncogenes, including Myc, Bcl-2, Pim-1, EGFR, β-catenin and Survivin. Molecules involved in cell cycle, EMT and cell adhesion are affected as well. The putative therapeutic relevance of SATB1 inhibition is further supported in an in vivo tumor xenograft mouse model, where the treatment with polymeric nanoparticles containing SATB1-specific siRNAs exerts antitumor effects. Conclusion: Our results demonstrate that SATB1 may represent a promising target molecule in glioblastoma therapy whose inhibition or knockdown affects multiple crucial pathways

Vergleich schneller, einfacher und robuster Extraktionsmethoden für die Qualitätskontrolle von Thymian

Comparison of fast, easy, and robust extraction methods for quality control of thym

Modulation of GLO1 expression affects malignant properties of cells

The energy metabolism of most tumor cells relies on aerobic glycolysis (Warburg effect) characterized by an increased glycolytic flux that is accompanied by the increased formation of the cytotoxic metabolite methylglyoxal (MGO). Consequently, the rate of detoxification of this reactive glycolytic byproduct needs to be increased in order to prevent deleterious effects to the cells. This is brought about by an increased expression of glyoxalase 1 (GLO1) that is the rate-limiting enzyme of the MGO-detoxifying glyoxalase system. Here, we overexpressed GLO1 in HEK 293 cells and silenced it in MCF-7 cells using shRNA. Tumor-related properties of wild type and transformed cells were compared and key glycolytic enzyme activities assessed. Furthermore, the cells were subjected to hypoxic conditions to analyze the impact on cell proliferation and enzyme activities. Our results demonstrate that knockdown of GLO1 in the cancer cells significantly reduced tumor-associated properties such as migration and proliferation, whereas no functional alterations where found by overexpression of GLO1 in HEK 293 cells. In contrast, hypoxia caused inhibition of cell growth of all cells except of those overexpressing GLO1. Altogether, we conclude that GLO1 on one hand is crucial to maintaining tumor characteristics of malignant cells, and, on the other hand, supports malignant transformation of cells in a hypoxic environment when overexpressed