Cells that cover the body’s inner and outer surfaces are called epithelial cells. Endothelial cells are specialised epithelial cells which, among other things, line the inside of blood vessels. The endothelium is anchored to the basement membrane through molecules called laminins. In an acute inflammation laminins can bind to leukocytes so that they can reach the inflamed tissue. Chemokines are molecules that attract leukocytes and can be synthesized by endothelial cells. This report will investigate what impact stimulation with laminin-332 on endothelial cells has on their gene expression for the chemokines CCL7, CCL8, CCL20, CXCL6 and CXCL10. A previously performed analysis for protein expression which had been performed under the same conditions revealed an upregulation of all chemokines except for CCL8, which was downregulated. The analysis for protein expression was executed with Olink’s Proximity Extension Assay and analysis of gene expression was carried out with qRT-PCR. The results revealed that gene expression for CCL8, CXCL6 and CXCL10 was under the detection limit for the chosen method. Gene expression for CCL7 and CCL20 was detectable and revealed an upregulation of gene expression for both genes, which was consistent with the results from the study that analysed protein expression. This led to the conclusion that stimulus with laminin-332 upregulates mRNA expression, protein production and protein secretion in human umbilical vein endothelial cells for chemokines CCL7 and CCL20. Lastly, the involvement of the chemokines CCL7 and CCL20 in inflammation and cancer diseases is explored as well as their potential role as a biomarker for clinical treatment. / Celler som täcker kroppens inre och yttre ytor kallas för epitelceller. Endotelceller är specialiserade epitelceller som bland annat bekläder insidan av blodkärlen. Endotelet är förankrat till basalmembranet via molekyler kallade lamininer. Vid en akut inflammation kan lamininer binda till leukocyter för att de ska kunna ta sig ut till den inflammerade vävnaden. Kemokiner är molekyler som attraherar leukocyter och som kan produceras av endotelcellerna. I denna rapport utforskas vilken påverkan som laminin-332 har på endotelcellers genuttryck för kemokinerna CCL7, CCL8, CCL20, CXCL6 och CXCL10. En tidigare utförd analys för proteinuttryck som gjorts under samma förhållanden visade en uppreglering av samtliga kemokiner, med undantag för CCL8 som blev nedreglerad. Analysen för proteinuttryck var utförd med Olinks Proximity Extension Assay och analys för genuttryck utfördes med qRT-PCR. Resultaten visade att genuttrycket för CCL8, CXCL6 och CXCL10 var för lågt för att detekteras med den valda metoden. Genuttryck för CCL7 och CCL20 var detekterbart och visade båda en uppreglering av genuttryck vilket överensstämde med resultatet från studien som analyserat proteinuttrycket. Detta ledde till slutsatsen att stimulans med laminin-332 uppreglerar uttryck av mRNA, proteinproduktion och proteinsekretion i humana venösa endotelceller från navelsträng för kemokinerna CCL7 och CCL20. Slutligen, utforskas involveringen av kemokinerna CCL7 och CCL20 vid inflammation och cancerassocierade sjukdomar samt vilken roll de kan spela som biomarkörer vid behandling.

No description available.

No description available.

No description available.

No description available.

The incidence of sepsis is a growing problem worldwide with a high mortality rate.  K- capsule antigens are becoming more dangerous than before. The purpose of the study was to categorize the capsules virulence factors, as well as to find a safe and empirical antibiotic treatment for sepsis infection, and to determine if the existence of ESBL producing bacteria have increased exponentially in recent decades. A total of 101 isolates were collected for a period of 5 years, of which blood isolates (n=38) were collected at Uppsala university hospital and feces isolates from healthy mothers and their infants (n=63) at Cunghi hospital, China. Isolates were serotyped with agglutination test, using N. meningitides B / Ecoli K1 against K1 capsule antigen, and phage test, using K1, K5 and K13 bacteriophage to identify the corresponding E. coli K antigen. Results showed that K1- capsule antigen could be identified in 42% (16/38) of the blood-isolated bacterial strains, compared with 11% (7/63) in fecal-isolated bacterial strains, while K5- capsule antigen was serotyped in 5% (2/38) and 19% (12/63), respectively. In contrast, the K13 capsule antigen was not found in blood-isolated bacterial strains and was serotyped only in 4% (3/63) of the fecal-isolated bacterial strains. Overall, the investigated E.coli  K capsular antigens were identified in 47% (18/38, non -typeable n=20) of the blood-isolated cultures compared with 35% ( 22/36, non-typeable=41) in fecal-isolated cultures..

Immuncytokemi (ICC) har blivit en framgångsrik metod inom cancerdiagnostiken. ICC bygger på antikropps- och antigenreaktion då infärgning med specifika antikroppar kan lokalisera strukturer hos celler. Hos klinisk patologi i Kalmar utförs ICC med den traditionella prepareringsmetoden där materialet formalinfixeras, bäddas i paraffin och snittas enligt cell block teknik (CBT). Vid ICC med vätskebaserad cytologi (LBC, liquid-based cytology) kan infärgning utföras på en mindre mängd material samt ge ett resultat 1-2 dagar innan resultat fås med CBT. ICC med LBC saknar kontroller vilket medför att resultatet blir mindre tillförlitligt. Syftet med examensarbetet var att ta fram positivt kontrollmaterial för ICC av LBC-preparerat material. Kontrollmaterial selekterades från tidigare diagnostiserade och immunfärgade patientprover i form av exsudat och tonsillmaterial. En panel av 6 monoklonala antikroppar bestående av anti-CD20, anti-CD3, anti-CK5, anti-CK7, anti-Ki-67 och anti-WT1 användes. Kontrollmaterialet preparerades enligt ThinPrep™-metoden och sattes till objektglas med Cytospin™. När kontrollmaterialets positivitet bekräftats testades det i rutinarbete ihop med 10 patientprover. Patientproverna preparerades med ThinPrep™-metod och placerades på objektglasen med hjälp av en ThinPrep™5000 processor. Samtliga resultat jämfördes med infärgning gjord med CBT och majoriteten av resultaten var likvärdiga. Ur resultatet drogs slutsatsen att metoden verkar lovande men prepareringssteg och färgprotokoll behöver optimeras samt att större studier med fler patientprover behövs för att kunna beräkna tillförlitlig statistik. / Immunocytochemistry (ICC) has become a successful method of cancer diagnosis. ICC is based on antibody and antigen response, where staining with specific antibodies can localize structures in cells. Today at clinical pathology in Kalmar ICC is performed with the traditional method of preparation where the material is formalin fixed, embedded in paraffin and then cut into thin sections called cell block technique (CBT). ICC with liquid-based cytology (LBC) can be performed on a smaller amount of material and give results 1-2 days before results are obtained with CBT. ICC combined with LBC lacks controls so the result is not reliable. Purpose of this essay was to produce positive control material for the ICC of LBC-prepared material. Control materials were selected from previously diagnosed and immune-stained patient samples consisting of exudate and tonsillar material. A panel of 6 monoclonal antibodies, anti-CD20, anti-CD3, anti-CK5, anti-CK7, anti-Ki-67 and anti-WT1, was used. Control material was prepared according to the ThinPrep ™ method and was added to slides with Cytospin ™. After confirming the positivity of the control material, it was tested in routine work with 10 patient samples. Patient samples were prepared using the ThinPrep ™ method and added to the slides with the ThinPrep ™ 5000 processor. Results compared to staining with CBT showed correlation. Conclusion was that the method seems promising but stages of preparation and staining protocols need to be optimized and larger studies with more patient samples must be done in order to get reliable statistic data.

Science curriculum and pedagogy are at the center of a centuries-long debate concerning the appropriate relationship of faith and science. The difficulties that science educators face seem to be based in misinformation about the historical roots of this conflict. To address that conflict, the goals of this research were to separate myth from reality and to provide a necessary context to the current tensions that are disrupting science pedagogy and curriculum content within American public schools. Working within a theoretical framework of historical literacy, this qualitative, historical analysis was a comprehensive examination of the relationship of faith and science from ancient times through the Renascence to the emergence and development of Darwinism. The historical approach methodology was utilized as a means to document the systematic examination of past events, in order to illuminate and interpret the meaning of those events. The historical record revealed that science and religion are not necessarily incompatible and that the early Christian religion provided a fertile environment in which modern science could emerge. Also noted were many instances where the record was inconsistent with what educators have commonly taught as historical fact. Finally, the complex sources of tension between modern fundamentalist Christianity and Darwinism, which has appeared as a flashpoint in public discourse within science education, were examined in depth. Based on this analysis, the study includes recommendations for educators in their approach to addressing these challenges and teaching science. This analysis can produce positive social change for educators and their students, as this information is advanced as a means to enhance historical literacy among educators and their students.

The mesenchymal stromal cell (MSC) is of great interest for its immunomodulatory and regenerative properties. However, to research and use these MSCs it is essential to identify and characterize them as such. They need to fulfill the MSCs' minimal criteria which assess the differentiation potential, cell surface markers, and adherence. In this study, cells donated from human bone marrow were identified as MSC according to the minimal criteria. Methods used were flow cytometry, immunofluorescent staining, and ELISA. Furthermore, the population was cultured in three different media (DMEM-LG with either 10% FBS, 2% FBS, or 10% FBS supplemented with 10% conditioned media from human urinary bladder carcinoma cells (T24)) for 21 days whereupon tested for the mesenchymal characteristics, cells were counted and size measured at every passage. All cultures maintained their mesenchymal character, however, cells grown in 2% FBS became a considerably more heterogenous population regarding cell size and granularity, perhaps because of senescence. Additionally, these cells somewhat decreased in proliferation and resulted in 1 x 106 cells after 21 days, however, this was not a significant decrease when compared to the 10% FBS culture which had 2.16 x 106 cells after 21 days (p=0.061). On the contrary, the culture supplemented with T24 conditioned media resulted in a significantly higher cell count with 4.75 x 106 cells (p=0.008). Further studies could investigate which components in the conditioned media contributed to the proliferation. Moreover, the cell population in this study could not be characterized as MSC with certainty as additional cell surface markers should be tested.

Immunocytokemi på utstryk är en metod för sortering av celltyper och identifiering av maligna celler. Metoden anses som ett aktuellt alternativ till den redan etablerade immunocytokemi på cellblock-metoden vid diagnostik av maligniteter. Syftet med examensarbetet är metodutvärdering och etablering av immunocytokemi på utstryk för sortering och identifiering av olika celltyper och maligna celler, samt metodjämförelse mellan utstryk och cellblock. Fem prover från pleuravätska framställdes till utstryk och cellblock. Immunocytokemi med antikropparna anti-Epithelial Specific Antigen, anti-Cytokeratin 7 och anti-Carcinoembryonic Antigen tillämpades på dessa. Resultatbedömning omfattade bevarandet av morfologin, färgupptag och ospecifik färgning hos celler och bakgrund, samt möjligheten att ställa säker diagnos för utstryk. Resultat visar färgupptag och viss bevarad morfologi hos utstryk. Samtliga utstryk har ospecifik färgning och kontamination som medför svårighet att urskilja celltyper och identifiera maligna celler. Möjlighet att ställa diagnos med utstryk varierar med preparaten. Färgningen hos samtliga cellblock är robust och mer specifik för celler och bakgrund. Examensarbetet når slutsatsen att etablering av immunocytokemi- metoden på utstryk är möjligt och bör tillämpas i vissa situationer. Etablering som rutinmetod är olämpligt. Framställning av preparat med hög kvalitet och specifika markörer är utvecklingsområden för laboratoriet och vidare forskning. / The method of immunocytochemistry on smears is utilized for categorizing cell types and identifying malignant cells. Immunocytochemistry on smears is a prospective alternative to the already implemented cell block method when diagnosing malignancies. The thesis aims to evaluate and implement immunocytochemistry on smears for categorizing and identifying cell types and malignant cells, along with comparing smears and cell blocks. Five pleural samples were processed into smears and cell blocks. Staining of these occurred with antibodies anti- Epithelial Specific Antigen, anti-Cytokeratin 7 and anti-Carcinoembryonic Antigen. Evaluated factors included preservation of morphology, stain uptake and non-specific staining for cells and background and finally the ability to generate a definitive diagnosis for smears. Results demonstrate stain uptake and preservation of morphology in some smears. All smears demonstrate non-specific staining and contamination, causing difficult categorization and identification. Generating a definitive diagnosis varies among samples. The thesis concludes that immunocytochemistry is a suitable platform for smears and a prospective implementation at the laboratory in certain situations. Immunocytochemistry on smears is not eligible as a routine method. Preparation of high-quality smears and specific antibody markers are deemed points of interest for the laboratory and future studies.