BackgroundAll cells carry a transmembrane proteoglycan calledsyndecan. Syndecans influence many functions like cell migration, cell adhesionand cell proliferation and it is involved in cellular signaling andtumourigenesis. The common features of differentiation in twomesenchymal tumor cell types, malignant mesothelioma cells and fibrosarcoma cells,are connected to the synthesis of syndecans. By studying the overexpression ofsyndecan-1 we hope to discover new features of the syndecan-1 molecule that wecan add to the puzzle of mesenchymal tumors. Methods and findingsMalignant mesothelioma cells and fibrosarcoma cellswere cultured and transfected with full-length- and truncated syndecan-1 constructs.To detect the expression of syndecan-1 on RNA level Rt-Q-PCR was conductedfollowed by immunocytochemical analysis to establish the syndecan-1 expressionon protein level. The result showed a 2-7 fold increase of syndecan-1 in thetransfectants comparing to the control. The proliferation of transfectants was analyzedby cell proliferation assay and cell cycle analysis. All transfectants showed alower proliferation rate comparing to the controls and a slight increase inG0/G1 phase. Because of the high structural similarities ofsyndecan family members, I studied how overexpression of syndecan-1 affected theother syndecans using Rt-Q-PCR. Syndecan-2 and -4 were downregulated in thetransfectants carrying syndecan-1 ectodomain, whereas the truncated versionshad the opposite effect. The expression of syndecan-bound heparan sulfate wasstudied by FACS and indicated an upregulation for heparan sulfate whenmeasuring internal- and membrane bound syndecans simultanesly. ConclusionsIn this study I haveshown that overexpression of full-length syndecan-1 and the different truncatedvariants, had similar profound effects on mesenchymal cell proliferation. Syndecan-1also influences the other members of the syndecan family suggesting a complexregulation.

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Many everyday tasks and also many sports require good grip and forearm strength. Everything from carrying boxes and lifting a child to grab the arms and legs in various martial arts, or holding a tennis racket involving the hand and forearm muscles in various ways. It is therefore important to train these muscles to prevent injuries and congestion and to perform well in sport. A well known way to train functional strength in the hand and forearm muscles is weight training with thick handles. The purpose of this study was to investigate the differences in muscle activity in the upper arm and forearm muscles as well as the deltoids between two bars of different diameter (28mm and 57mm) in two different weight training exercises using electromyography (EMG). The weight training exercises consisted of a pulling exercise in the form of a bench row and a pressing exercise in the form of close-grip bench press. The study also examines whether there is a connection between hand strength and muscle activity, and if there is any connection between hand size and muscle activity. Results show that muscle activity between the thick bars remained unchanged in the close-grip bench press. In the bench row exercise, however, significant increases were seen in the forearm flexors and m. biceps brachii while lifting the thicker bar. The forearm extensors showed an indication of muscle activity increases while lifting the thicker bar. However, there was no connection between hand strength and muscle activity and no correlation between hand size and muscle activity. The conclusion is that the pulling exercise with the thicker bar results in higher muscle activity in comparison to a standard Olympic bar in several muscles involved, not just those directly affected by the thicker bar.

Since the discovery in 1989 of hepatitis C virus (HCV) as the infectious agent responsible for the vast majority of post-transfusion non-A non-B hepatitis, blood transfusions are no longer a source for HCV transmission in Sweden. Anti-HCV testing was implemented for all blood donations in 1992. Since then intravenous drug use (IDU) has become the major route of transmission in the western world. Six genotypes and more than 80 subtypes of HCV have now been identified world-wide. These genotypes and subtypes are determined by genetic divergences between the HCV strains. Subtypes 1a, 1b, 2b, 2c, and 3a have global spread, while the other subtypes have a more limited geographical distribution. Little was known on the prevalence of HCV among blood donors and on which genotypes and subtypes of HCV were circulating in Sweden before the testing of all blood donations was implemented. The prevalence of anti-HCV was therefore investigated in sera sent to the Swedish Institute for Infectious Disease Control (SMI) from 412 patients; 241 were sampled between 1970 and 1991 before the universal screening in 1992, while 171 were sampled between 1992 and 2002. The samples derived from 193 (47%) blood donors, (104 sampled before, and 89 after 1992), and from seven other groups of patients. Two groups had suspected known routes of infection, intravenous drug use (IDU) 33 patients and hemodialysis, 16 patients, while it was unknown for the other patients. Anti-HCV was detected in 120 (29%) samples. The highest frequency was found among IDUs, (91%). Before general screening was implemented, 2.8% of the blood donors were positive for hepatitis C, whereas 28% of those sampled after 1992 were anti-HCV positive. Those latter samples were sent to SMI due to anti-HCV reactivity in a primary test at the blood centre. HCV RNA could be detected by PCR in 56 (47%) of the anti-HCV positive samples, the subtype could be determined by sequencing in 45 (80%) of those. The subtypes found were 1a in 31 %, 1b in 18%, 2b in 22%, and 3a in 27%. One sample was of subtype 2c. There was a tendency of increase of genotype 2 and a decrease in subtype 1a with time. 1a was found in 38% of the samples collected before 1992, while it was only found in 19% of the samples from 1992 or later. On the other hand genotype 2 was found in 17% sera sampled before 1992 and in 37% of the samples collected 1992 or later. It is not known if this genotype has recently been introduced into Sweden. Further analysis on larger series of samples is needed to confirm these preliminary results. / AcknowledgmentsI would like to express my gratitude to several people who have been supportive in different ways throughout this project.First of all, I want to thank my supervisor Helene Norder, for giving me the possibility to do my diploma thesis at the Department of Virology, Swedish Institute for Infectious Disease control (SMI) and for helping me during this study and for the many insightful conversations during the design and development stages of the application, and also for the many helpful comments and suggestions on the text of the thesis.I want to express my appreciation to my laboratory supervisor Regina Wallin, Camilla Jern and Josefine Ederth for helping me during the procedure for this study. Then, I want to thank my examiner Magnus Johansson from the Södertörns university collegefor his advice on writing this paper. Finally, I would like to thank my family and specially my mother Bahar Hamid for always supporting me during my whole life.Last, but not least, I would like to thank my friends Annika Andersson and Yourdons Yemane for being encouraging, understanding and always supportive.

Introduction The genetic skin disease epidermolytic ichtyosis is caused by mutations in either keratin gene 1 or 10 and leads to blisters and hyperkeratosis of the epidermis. At cellular level the disease is seen as aggregates in the keratin filaments. Since medicines are hard to investigate and produce mainly due to lack of reproducible model systems, there is no good treatment available for this disease today. In this article we describe how an in vitro model consisting of cells from a stable cell line transfected with expression plasmids to mimic patient cells, may be a possible alternative for screening compounds for therapies. The first step was to generate an expression plasmid required to complete the in vitro model. The model was tested by a preliminary experiment with 4-phenylbuturate (4-PBA) to see if the substance had an effect on the amount of cells with keratin aggregates. Methods PCR and primers containing the desired mutation were used to incorporate a deletion in wild type keratin gene1 plasmid to generate the expression plasmid. HaCaT cells were transfected with the plasmid for expression of keratin. The percentage of cells with keratin aggregates was assessed by fluorescence microscopy. Results/Conclusion Cells containing mutated plasmid had a higher percentage of keratin aggregates compared to cells transfected with wild type plasmid. 4-PBA was found not to affect the amount of cells with keratin aggregates. According to this project the model might be a useful tool for screening compounds, but it needs to be more developed and tested.

Prostaglandin (PG) E2 is an eicosanoid derived from the polyunsaturated twenty carbon fatty acid arachidonic acid (AA). PGE2 has physiological as well as pathophysiological functions and is known to be a key mediator of inflammatory responses. Formation of PGE2 is dependent upon the activities of three specific enzymes involved in the AA cascade; phospholipase A2 (PLA2), cyclooxygenase (COX) and PGE synthase (PGEs). Although the research within this field has been intense for decades, the regulatory mechanisms concerning the PGE2 synthesising enzymes are not completely established. PGE2 was investigated in human monocytes with or without lipopolysaccharide (LPS) pre-treatment followed by stimulation with calcium ionophore, opsonised zymosan or phorbol myristate acetate (PMA). Cytosolic PLA2a (cPLA2a) was shown to be pivotal for the mobilization of AA and subsequent formation of PGE2. Although COX-1 was constitutively expressed, monocytes required expression of COX-2 protein in order to convert the mobilized AA into PGH2. The conversion of PGH2 to the final product PGE2 was to a large extent due to the action of microsomal PGEs-1 (mPGEs-1). In addition, experiments with inhibitors of extracellular signal regulated kinase and p38 activation, indicated that phosphorylation of cPLA2α was markedly advantageous for the formation of PGE2. Ellagic acid, a natural polyphenolic compound found in fruits and nuts, was shown to inhibit stimuli induced release of PGE2 in human monocytes. The effect of ellagic acid was not due to a direct effect on the activities of the enzymes but rather to inhibition of the LPS-induced protein expression of COX-2, mPGEs-1 and cPLA2a.

Abstract G-protein coupled receptors (GPCRs) are having the high medical importance since almost half of the medicinal drugs are designed as modulators of receptor molecules. Crystal structure or NMR structures of GPCRs are very difficult to determine because all GPCRs are typically bound to the cell membrane and thus their molecular activation mechanism is still unclear. The recent publication of the crystal structure of the 2-adrenoreceptor will provide new insights in the field of GPCR research. Ghrelin is a peptide growth hormone which binds to the growth hormone secretagogue receptor (GHS-R) and stimulates the release of growth hormone. Based on the known ghrelin receptor binding core sequences wFwLL (upper letter and lower letter representative for L-form and D-form of the amino acids respectively), we prepared two novel peptide analogs with terminal S-(2-aminoethylsulfenyl) cysteine residues. These peptides were tested for their ability to suppress the binding of ghrelin to transfected COS7 cell-line (Kidney fibroblast line from the green African monkey) cells expressing the ghrelin wild-type receptor or certain mutants thereof. As a result we observed a significant reduction of the total number of binding sites accessible for ghrelin, which increased with the time the cells were incubated with our test compounds. This observations support our hypothesis that the peptides we tested form a covalent bond with free thiols located closely to the ligand binding-site of the receptor protein by disulfide thiol exchange which is an interesting target for development of anti-obesity drugs.

ABSTRACT In the western world colitis is a common chronic disease and in Sweden the prevalence is around 1%. If a patient has bloody diarrhea it is probably ulcerative proctocolitis or Crohn’s disease, whereas if the diarrhea is watery, it is microscopic colitis. For a diagnosis, the patient has to do a colonoscopy and a colonic biopsy sample has to be taken. The biopsy sample will be sent to a laboratory for sectioning, staining and microscopic analysis. In this study we compared the effects of short and long dehydration time of the sample before the sectioning. We also compared staining with Alcianblue/Van Gieson and Van Gieson alone. Our results showed that a short dehydration time was a milder treatment and made it easier to section the biopsy sample. The comparison of the two methods was unsuccessful because the staining with Alcianblue/Van Gieson failed.

Background. Immunohistochemistry is a useful method in the differential diagnosis between pleural mesotheliomas and metastatic adenocarcinomas in the pleura. Cytokeratin 20 and 7 have been used successfully as markers in studies determining primary location of adenocarcinomas from metastases. The current study is a complementary research of archived paraffininbedded material of cases with cancer origin. This study contributes a bigger statistical material that may facilitate the search for unknown primary site of adenocarcinoma by identification of metastatic cells in the pleura. Methods. Cells from the pleura taken from fifteen patients with diagnosed cancer of different types and eleven patients with cancer of unknown origin, were stained with antibodies against the tumour markers: Ber EP 4, calretinin, cytokeratin 20 and 7, estrogen receptor α, thyroid transcription factor, prostate-specific antigen and Cdx2.The staining was conducted in an automated immunohistochemical system. The staining of each kind of antibody was confirmed by a control section staining. Results. All control staining ended perfect The whole panel of antibodies used on mammary cancer showed the same pattern for every antibody. Of the patients with cancer of unknown origin there were four that gave the same pattern, two men and two women. The women are deceased. To make a more careful evaluation more information and clinic background is needed. The number of samples is too small to draw any statistical conclusions. Comment. Although the control staining was perfect the negative result of CK20 in the cases of diagnosed colon cancer was unexpected. This staining should be performed again to confirm the result. In some cases the number of cells were to few for a certain evaluation. The slides and the results of this work will be archived for further research.

The human skin provides us with an excellent protective barrier and possesses a remarkable ability of constant renewal. Basal cell carcinoma is the most common type of skin cancer. The aim of this project was to verify results from an earlier study investigating the molecular differences between basal cell carcinoma (BCC) and basal cells of normal human epidermis. In that study microdissection of cell populations from BCC and basal cells of normal epidermis respectively was performed in five cases of confirmed BCC. Following RNA extraction and amplification, a gene expression analysis was performed using a 46 k human cDNA microarray. Comparison of expression profiles showed a differential expression of approximately 300 genes in BCC. An upregulation of signaling pathways previously known to be of importance in BCC development could be observed, as well as a downregulation of differentiation markers, MHC class II molecules, and proteins active in scavenging of oxygen radicals. We wanted to confirm these findings for a number of selected genes, using real time PCR. The focal point of this project was microdissection of cells from BCC and subsequent isolation of RNA. Microdissection based methods offer a possibility of selecting well defined cell populations for further analysis by using a focused laser beam. Initially tests in order to optimize the method were also performed, concerning the dehydration process and choice of slides used in microdissection. Isolation of RNA may, as we experienced, be associated with problems due to destruction of RNA by degrading enzymes.