Pathogen burden and cortisol profiles over the day

Hypothalamic-pituitary-adrenocortical (HPA) regulation in adults is influenced by early psychosocial adversity, but the role of infectious disease history is poorly understood. We studied the association between cumulative pathogen burden and cortisol profile over the day in a sample of 317 healthy men and women aged 51-72 years. Cumulative pathogen burden was defined as positive serostatus for Chlamydia pneumoniae, cytomegalovirus (CMV) and herpes simplex virus 1 (HSV-1). Salivary cortisol was sampled repeatedly over the day. The cortisol slope was defined as the decrease across the day and evening. Age, gender, grade of employment, body mass index, smoking status, self-rated health, cardiovascular medication, depressed mood and time of waking were included as covariates. The pathogen burden averaged 1.76 (S.D. = 0.92). The cortisol slope was inversely associated with pathogen burden after controlling for covariates. When individual pathogens were studied, only CMV was associated with flatter cortisol rhythms in isolation. We conclude that pathogen burden is independently associated with flatter cortisol slopes over the day, and may contribute to disturbed neuroendocrine regulation

Metadata i Cross Media Redaktionsprocesser

Olika typer av media är inte längre bundna till specifika distributionskanaler i samma grad som de en gång varit. Nya krav på mediainnehåll samt -tillgänglighet har lett till att företag inom mediabranschen måste anpassa sig genom att tillämpa nya rutiner för skapande och administrering av innehåll. Detta inverkar inte endast på arbetsrutiner, utan leder i sin tur till nya krav på DAMS system i vilka metadata kommer inneha en väsentlig roll. Utan välstrukturerad metadata kommer många arbetsuppgifter visa sig extremt tidskrävande. Den här studien erbjuder en genomförlig och aktuell empirisk forskning med insyn på tillämpningen av metadata inom innehållsproduktionen hos välkända medieföretag som utövar cross media publicering. Företag som deltog i studien kan beskrivas som nyhetsbyråer, tidnings- eller tidsskriftsutgivare. En abstrakt metadatastruktur bestående av essens, kontextuell, strukturell, och administrativ metadata var konstruerad på basen av uptäckt metadata hos de besökta företagen. Denna struktur avspeglar nuvarande krav på metadata inom cross media företag. Metadatastandarder rekommenderas som bas för kartläggning mellan lämpliga metadatafält och de i strukturen angivna metadata-attributen. På basis av resultaten gavs även förslag på framtida krav på metadata inom cross media redaktionssystem. Metadata var även analyserad på basen av typ, egenskaper, och användning. Subjektiva åsikter om nuvarande metadatavanor togs även i beaktande. Redaktionsprocesser inklusive metadatalivscykeln var modellerade med BPMN. Bland de företag som deltog i studien uppkom skillnader i processer anknytna till cross media publicering. Alla deltagande parter visade dock ett gemensamt intresse för att förbättra den nuvarande situationen. En annan gemensam överenskommelse uppstod i formen av ett framtida krav på redaktionssystem gällande temaeller ämnescentrerad planering av innehåll. I synnerhet essensbaserad metadata är i starkt behov av förbättringar för att klara av framtida krav. Medieföretag har insett att investeringar i metadata inte endast minskar på arbetsbördan och är till förmån för administrering av tillgångar, utan också erbjuder totalt nya möjligheter för affärsverksamhet.Different types of media such as text, images, audio, and video are no longer restricted to different publishing channels in the same way that they used to be. Media companies are adopting new routines for handling and creating content to better suit the new demands, which in turn puts new requirements on digital asset management systems (DAMS). Metadata will have an essential role in managing content in these systems, and without well structured metadata many tasks become extremely time consuming. This study provides an extensive, up-to-date, empirical research of the current use of metadata across the content creation process within well known media companies involved in cross media publishing. Three case companies were selected for this study: a newspaper, a magazine publisher, and a news agency. Observations and surveys, including interviews and questionnaires were conducted at the research sites. A metadata framework consisting of essence, contextual, structural, and administrative metadata field attributes was created based on the discovered metadata in case companies. The framework serves as a platform for current metadata needs in media companies utilizing cross media publishing. Metadata standards are suggested to be used as a source for mapping suitable metadata fields to the provided metadata attributes. Suggestions for future requirements on metadata in editorial systems were also given based on the results. In addition to the metadata framework, discovered metadata was also assessed based on nature, characteristics, and use. Subjective viewpoints of current metadata practices are also taken into account. Current editorial processes including the metadata time-cycle were modeled with BPMN annotation language. There were differences in cross media publishing routines between the case companies, but all participants showed a common interest to enhance current practices. A mutual requirement on future editorial systems was the inclusion of theme or topic based planning of content. The results showed that especially essence based metadata needs improvements in order to cope with future requirements. Media companies have realized that investing in metadata not only reduces workload and is beneficial from an asset management perspective, but also provides completely new business opportunities

New insights into the genetic basis of colorectal cancer

Colorectal cancer (CRC) is the third most common cancer, and the second most common cause of cancer mortality. The aim of this thesis work was to gain novel insight into the molecular mechanisms behind CRC predisposition, as well as tumor progression and development. Microsatellite instability (MSI) arises due to a defective mismatch repair system and is characteristic for a subset of all CRCs. Here, we aimed to identify novel MSI target genes with potential oncogenic effects. We characterized all genes overexpressed in MSI CRCs and predicted to escape nonsense-mediated decay when mutated. The mitotic checkpoint kinase TTK was identified with protein-elongating mutations in 59% of MSI CRCs. TTK has an essential role in spindle assembly checkpoint (SAC) signaling, however, the mutated protein did not show SAC weakening. While no evidence of oncogenic mechanisms was observed, the high mutation frequency of TTK argues for biological significance. In another screening effort on MSI CRCs, we sought to identify novel oncogenes with activating hotspot mutations. The exomes of 25 tumor and respective healthy colon tissues were sequenced. A total of 15 candidate oncogenes with hotspot mutations were identified. Three genes, ZBTB2, PSRC1 and RANBP2, displayed hotspot mutations also in the validation set of 86 MSI CRCs. Interestingly, the CRC-associated mutant form of ZBTB2 increased cell proliferation. Additional work is needed to further clarify the role of the identified somatic mutations in CRC tumorigenesis. The candidate oncogenes identified in this thesis work might be used to develop personalized tumor profiling and therapy. Inherited susceptibility is estimated to be involved in approximately one-third of all CRCs. However, the great majority of inherited CRC susceptibility remains still molecularly unexplained. A recent systematic sequencing study on CRC reported a set of somatically mutated genes, termed candidate cancer (CAN) genes. We examined the mutational profiles of 15 top-ranked CAN genes for somatic mutations as well as for germline variants in 45 familial CRC cases. In our tumor set, six of the CAN genes were somatically mutated. In germline, three private missense variants were identified in CSMD3, EPHB6 and c10orf137. In another effort, we exome sequenced 96 independent cases with familial CRC. We identified 11 novel candidate CRC susceptibility genes with rare putative LoF variants. Seven loss-of-heterozygosity events, involving four genes, were observed in the data. In each occasion, the losses targeted the wild-type allele (P=0.0078), providing further support that true culprits are among the eleven genes. This study provides an interesting set of candidate predisposing genes, which might explain a subset of common familial CRC. The identified germline variants need to be validated in larger sample sets to provide firm evidence for disease predisposition. Additional work is also needed to characterize the detailed functional and clinical relevance of the identified candidate CRC predisposing genes. This information, then, can ultimately be translated into tools for cancer prevention and early diagnosis in individuals carrying true predisposition alleles.Kolorektal cancer (KRC, cancer i tjock- och ändtarm) är en av de vanligaste cancerformerna i västvärlden. Flera faktorer, både ärftliga och i människans omgivning, påverkar uppkomsten av KRC. En kartläggning av de genetiska förändringarna bakom KRC är viktig för att bättre förstå tumörutvecklingen. Sådan information kan sedan användas för att utveckla effektivare behandlingsmetoder för att minska död och lidande orsakade av KRC. De två huvudsakliga syftena i detta doktorandprojekt var att 1) få ny insikt i de molekylära mekanismerna bakom kolorektal tumörutveckling 2) identifiera nya genetiska faktorer som påverkar ärftlig benägenhet för KRC. En del kolorektala tumörer hör till gruppen mikrosatellit instabila (MSI). Dessa tumörer har ett defekt DNA-reparationssystem vilket leder till ett högt antal mutationer i DNA-sekvensen, speciellt i repetitiva DNA-segment. Syftet med detta projekt var att identifiera nya onkogener i denna tumörtyp. Onkogener är gener som ändrar grundläggande egenskaper hos celler, till exempel tillväxt och differentiering. Genetiska avvikelser i en sådan gen kan leda till malign transformation av cellen. Två olika DNA-sekvenseringsmetoder användes; den så kallade Sanger-dideoxy-metoden samt nästa generations DNA-sekvensering. Ett antal nya onkogener med hög mutationsfrekvens identifierades i kolorektala tumörer av gruppen MSI. Ytterligare studier krävs för att klarlägga hur dessa onkogener påverkar malign transformation av cellen. De identifierade onkogenerna kan möjligtvis användas för att utveckla mer effektiva strategier för individuell tumörprofilering och cancerterapi. Det finns klara bevis för att genetiska faktorer predisponerar för KRC i ungefär vart tredje fall. Ett antal kända ärftliga syndrom medför en ökad risk för KRC. Men dessa syndrom är sällsynta och bidrar bara till en bråkdel av fallen. Än i dag förblir en stor del av den ärftliga benägenheten för KRC molekylärt oförklarad. Det andra syftet med denna avhandling var att identifiera nya gener som ökar benägenheten till familjär KRC (patienter med en förstagradenssläkting med KRC). Även här användes Sanger-dideoxy-metoden samt nästa generations DNA-sekvensering. Flera attraktiva kandidatgener identifierades med protein-kapande DNA-variationer i patienternas könsceller. DNA-variationerna var väldigt sällsynta eller frånvarande från den allmänna populationen. Ytterligare forskningar, med större sampelmaterial, är dock nödvändiga för att bevisa att de identifierande DNA-variationerna påverkar den ärftliga benägenheten till familjär KRC. Identifikationen av nya gener bakom ärftlig KRC möjliggör en förbättrad patientförsörjning med tidig diagnos och förbättrad riskprofilering

Red Fluorescent Chlamydia trachomatis Applied to Live Cell Imaging and Screening for Antibacterial Agents

In this study, we describe the application of a transformed Chlamydia trachomatis strain constitutively expressing the red fluorescent protein mCherry, to allow real-time monitoring of the infection cycle and screening for agents that block replication of C. trachomatis. The red fluorescent C. trachomatis strain was detected autonomously without antibody staining and was equally susceptible to doxycycline as the wild type strain. A high-throughput screening assay was developed using the transformed strain and automated fluorescence microscopy. The assay was used in a pilot screen of a 349 compound library containing natural products from Australian flora and fauna. Compounds with anti-chlamydial activity were tested for dose response and toxicity to host cells and two non-toxic compounds had 50% effective concentration (EC50) values in the low micromolar range. Natural products are valuable sources for drug discovery and the identified Chlamydia growth inhibition may be starting points for future drug development. Live cell imaging was used to visualize growth of the red fluorescent C. trachomatis strain over time. The screening assay reduced workload and reagents compared to an assay requiring immunostaining and could further be used to monitor the development of Chlamydia inclusions and anti-chlamydial effect in real time

Differential interaction of glimepiride and glibenclamide with the β-cell sulfonylurea receptor I. Binding characteristics

Glimepiride is a novel sulfonylurea drug for treatment of non-insulin-dependent diabetes mellitus with higher blood sugar lowering efficacy in diabetic patients than glibenclamide raising the question whether this characteristics is in line with different binding of glimepiride and glibenclamide to the β-cell sulfonylurea receptor. Scatchard plot analysis of [3H]sulfonylurea binding to membranes isolated from rat β-cell tumors and (RINm5F) insulinoma cells and to RINm5F cells demonstrated that glimepiride has a 2.5–3-fold lower affinity than glibenclamide. This corresponded well to the 8–9-fold higher koff and 2.5–3-fold higher kon rates of glimepiride compared to glibenclamide as revealed by the dissociation and association kinetics of [3H]sulfonylurea binding and the Kd values calculated thereof. In agreement, the concentrations required for half-maximal displacement of [3H]sulfonylurea bound to β-cell membranes were significantly higher for glimepiride compared to glibenclamide. However, the binding affinity of glimepiride measured by both equilibrium binding and kinetic binding studies upon solubilization of β-cell tumor membranes and RINm5F cell membranes increased up to the value for glibenclamide. This was primarily based on a drastic decrease of the dissociation rate constant of glimepiride whereas the kinetics of glibenclamide binding remained largely unaffected upon solubilization. These data suggest that the Kd value alone is not sufficient for characterization of a sulfonylurea drug, since the kinetic binding parameters may also determine its acute blood sugar lowering efficacy

3 '-UTR poly(T/U) repeat of EWSR1 is altered in microsatellite unstable colorectal cancer with nearly perfect sensitivity

Approximately 15 % of colorectal cancers exhibit instability of short nucleotide repeat regions, microsatellites. These tumors display a unique clinicopathologic profile and the microsatellite instability status is increasingly used to guide clinical management as it is known to predict better prognosis as well as resistance to certain chemotherapeutics. A panel of five repeats determined by the National Cancer Institute, the Bethesda panel, is currently the standard for determining the microsatellite instability status in colorectal cancer. Recently, a quasimonomorphic mononucleotide repeat 16T/U at the 3' untranslated region of the Ewing sarcoma breakpoint region 1 gene was reported to show perfect sensitivity and specificity in detecting mismatch repair deficient colorectal, endometrial, and gastric cancers in two independent populations. To confirm this finding, we replicated the analysis in 213 microsatellite unstable colorectal cancers from two independent populations, 148 microsatellite stable colorectal cancers, and the respective normal samples by PCR and fragment analysis. The repeat showed nearly perfect sensitivity for microsatellite unstable colorectal cancer as it was altered in 212 of the 213 microsatellite unstable (99.5 %) and none of the microsatellite stable colorectal tumors. This repeat thus represents the first potential single marker for detecting microsatellite instability.Peer reviewe

Screening of Finnish RAD51C founder mutations in prostate and colorectal cancer patients

Peer reviewe

Methyl sulfonamide substituents improve the pharmacokinetic properties of bicyclic 2-pyridone based:Chlamydia trachomatis inhibitors

Chlamydia trachomatis infections are a global health problem and new approaches to treat C. trachomatis with drugs of high specificity would be valuable. A library of substituted ring fused 2-pyridones has been synthesized and evaluated for their ability to attenuate C. trachomatis infectivity. In vivo pharmacokinetic studies were performed, with the best candidates demonstrating that a C8-methylsulfonamide substituent improved pharmacokinetic properties important for oral administration. C8-Methyl sulfonamide analogue 30 inhibited C. trachomatis infectivity in low micromolar concentrations. Further pharmacokinetic evaluation at an oral dose of 10 mg kg(-1) showed an apparent bioavailability of 41%, compared to C8-cyclopropyl and -methoxy analogues which had negligible oral uptake. In vitro ADME (absorption, distribution, metabolism and excretion) testing of solubility and Caco-2 cell permeability revealed that both solubility and permeability is greatly improved with the C8-methyl sulfonamide 30, effectively moving it from BCS (Biopharmaceutical Classification System) class IV to II