Uppsatsens syfte är att pröva doktrin JP 3-24 mot en teoretisk grund samt hur underrättelseproblematik kopplat till upprorsbekämpning i Afghanistan relaterar till doktrinen. I syftet ingår också att koppla problematiken till svensk relevans då Sverige lutar sig mycket på amerikanska erfarenheter i underrättelsearbetet.   Arbetet utgår från en bred teorigrund som operationaliseras i ett analysverktyg där fyra dimensioner ur ett befolkningscentrerat perspektiv genomlyser empirin. Empirin är ett subjektivt urval med exempel från Afghanistan mellan 2006-2012. Resultatet visar att trots en teoretiskt och doktrinellt stabil bas så fokuserar fortfarande många amerikanska förband felaktigt avseende underrättelsearbetet. Relationen mellan teori-doktrin-praktik påvisar en tydlig korrelation i att om doktrinen följs maximeras sannolikheten för seger i upprorsbekämpning. Övergripande slutsats är att det krävs ett omtag gällande utbildning och förberedelse av förbanden där resurser bör styras om till kulturell förståelse, bättre databassystem, metodöversyn, analytikerutbildning, samordning av informationsintegrering samt en mer utvecklad underrättelseledning. / The purpose of this thesis is to examine the JP 3-24 doctrine against a theoretical basis and investigate how the intelligence problems of counterinsurgency inAfghanistanrelate to the doctrine. The aim is also to link the problem to its relevance forSwedensinceSwedenleans heavily onU.S.experience in intelligence matters.The thesis is grounded on a broad theoretical basis which is operationalized in an analysis in which five dimensions of a people-centered perspective are applied to the empirical data. The empirical data is a subjective selection of examples fromAfghanistanbetween 2006-2012.The result shows that despite JP 3-24’s theoretical and doctrinal solid foundation, manyU.S.units are still improperly focused. The relationship between theory, doctrine and practice shows a clear correlation between following the doctrine and maximizing the probability of victory in counterinsurgency. The overarching conclusion is that a new approach to the training and preparation of units is required, where resources should be redirected to cultural understanding, better database systems, method revision, analyst training, coordination of information integration and intelligence management and leadership.

Denna uppsats undersöker vilken påverkan som olika maktaspekter har på förskolebarnet. Studiens syfte är att ur fyra av förskolans policydokument utröna hur förskolebarnet konstrueras. Det teoretiska perspektiv som har använts är en verktygslåda som är inspirerad av Foucaults tankar angående makt: disciplinär makt, biomakt, diskurs och genealogi. Det är fyra olika policydokument som har undersökts: Barnstugeutredningen (del 1), Pedagogiskt program för förskolan, Lpfö 98 och Lpfö 18. För att visa hur det normala och patologiska barnet konstrueras i förskolan har dokumenten analyserats med ett diskursanalytiskt förhållningssätt som tydligt involverar Foucaults maktperspektiv. Diskursanalysen visar vilka egenskaper som anses vara önskvärda för förskolebarnen att internalisera, vilka diskurser som är synliga och vilken påverkan olika historiska händelser har på konstruktionen av barnet. Analysens resultat visar att det inträffar olika diskursförskjutningar beroende på hur samhällsstrukturen ser ut i det givna sammanhanget, vilket också har en påverkan på hur barnet konstrueras. Det normala barnet konstrueras efter de normer och värderingar som förespråkas i den rådande diskursen. Det patologiska barnet anses framför allt vara barn med annan kulturell bakgrund som inte lever upp till de förespråkade normerna. En reflektion av resultaten visar att den allt snabbare samhällsutvecklingen, där de sociala klyftorna ökar, bidrar till en situation i förskolan som är svårhanterlig och komplex. En slutsats som dras är att det är av vikt för barnen att förskolan är likvärdig, och för att förskolan ska bli likvärdig bör vissa punkter i läroplanen revideras. / <p>Examinator har godkänt att fulltexten byts ut på grund av en smärre justering som inte påverkar uppsatsens innehåll. Gävle 12 augusti 2020.</p>

Inom klinisk patologi bedöms vävnadsprover under mikroskop, med syftet att fastställa diagnos. Utgångsmaterial är antingen ett helt organ eller en liten bit från det sjukligt förändrade organet. I det senare fallet kallas preparatet för biopsi och dessa tas med hjälp av biopsiverktyg och oftast under ultraljudledning. Njurbiopsier kan tas vid misstänkt transplantatavstötning, av medicinska skäl, vid inflammatoriska tillstånd (nefriter) samt nefrotiska tillstånd. En del av biopsimaterialet fixeras i 4 % formaldehyd för ljusmikroskopisk (LM) bedömning, medan resten transporteras i transportlösningar som Michels- eller Zeus lösning. Det materialet fryses ned direkt för kryostatsnittning, dessa transportlösningar syftar till att bevara vävnaden färsk för immunofluorescens (IF) analys. Problem kan uppstå när materialet som bedöms med LM inte innehåller ett acceptabelt antal glomeruli, där minst 12- 15 stycken krävs för att biopsin skall vara representativ. Om det är alltför få till antalet brukar man tina och formaldehydfixera IF materialet. Morfologin blir oftast dålig och inte lämplig för bedömning. Den enda som man kan göra är att ombiopsera patienten. Detta är kostsamt, obekvämt för patienten och inte helt riskfritt. Syftet med den här studien var att undersöka ifall man kan ersätta transportlösningarna med ett fixativ som skulle medföra bättre morfologi i de fall som IF materialet måste tinas upp och användas för LM, men samtidigt erbjuda samma kvalitet på IF-signalen. Tre fixeringsmedel jämfördes med rutinmetodens 4 % formaldehyd; dels kommersiellt tillgängliga som Histochoice och HOPE (Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect) och ett fixativ som är baserat på zinkjoner. Resultatet visade att när det gäller LM-undersökning på paraffininbäddat material så gav HOPE och Histochoice acceptabel morfologi av glomeruli. Vid upptinande av materialet följt av formaldehyd/paraffin, sågs fortsatt god kvalitet i den glomerulära morfologin, medan den tubulära morfologin var klart undermålig. Zn-fixering ledde till bubblor i cytoplasman vilket inte är acceptabel morfologi. I IF-analys gav alla fixativ negativ signal utom Zn-fixering, vilken gav en positiv signal. Det innebär att ingen säker diagnos kan ställas vid användning av de undersökta fixeringsmedlen som kan ersätta transportlösningar eller rutinlaboratoriet formaldehyd. Zn-fixativ såg lovande ut om det bortses från bubblorna, men detta behöver undersökas ytterligare i framtida studier / lt is important for clinical pathology to assess tissue samples (preparations) under the microscope to make a diagnosis. Starting materials is either a whole organ, or a small piece of the diseased body. The material can also be in the form of so-called biopsies, which are taken by biopsytools and ultrasound. Kidney biopsy may be taken for suspected transplant rejection, medical reasons and at nephritic (inflammation), and nephrotic states. Half of the material from kidney biopsies are transported in transport solutions of the type Michel / Zeus that serves the purpose of preserving the tissue fresh for immunofluorescence (IF) analysis. The other half is fixed in 4% formaldehyde for light microscope (LM) assessment, a procedure that precludes IF staining due to destruction of the antigenicity of the tissues. Problems might occur when the material is assessed by LM and do not contain an acceptable number of glomeruli, at least 12-15 are regarded as acceptable for diagnosis. If these are fewer in numbers, the material previously used for IF may be thawed and formaldehyde fixed. The morphology is however often adversely affected by this procedure, rendering the tissue unsuitable for morphological assessment. Therefore the only remaining option is to re-biopsy patients. This is costly, inconvenient for the parients, and not completely safe. This study was performed in order to explore if transport solutions can be replaced by a fixative that would result in a better morphology after thawing of the tissues, but with retained IF -quality. Three fixing agents were compared with routine method 4% formaldehyde; the commercially available Histochoice and HOPE fixatives (Hepes-Glutamic acid buffer mediated Organic solvent Protection Effect) and a fixative that was based on zinc ions. The results showed that in terms of LM, the fixatives HOPE and Histochoice gave acceptable morphology of the glomeruli, while the tubules were poor. It also showed that there were a lot of bubbles in the cytoplasm at the Zn-fixing. As for IF-analysis, all were negative except Zn that gave a positive signal. This means that no secure diagnosis can replace the routine transport formaldehyde. Zn fixative looked promising, except the bubbles, but needs to be further investigated in future studies.

Akut hjärtinfarkt är globalt sett en vanlig dödsorsak. Hjärtinfarkter behöver inte ge tydliga symptom och dödsorsaken kan därmed vara okänd fram till en klinisk obduktion och att histopatolgiska studier av hjärtmuskelvävnad genomförs. Fosforwolframsyra-hematoxylin (PTAH) är en färgningsmetod som kan användas vid visualisering av hjärtinfarkt genom att färga myokardceller, fibrin, kontraktionsband och cellkärnor blå medan kollagen färgas rosa till rödbrunt. Syftet med examensarbetet var att optimera PTAH-färgningsprotokoll för att underlätta diagnosticering av hjärtinfarkt. Tre olika färgningsprotokoll, två med PTAH med Mallory Bleach (#1 och #2) och ett med PTAH med refixering i Bouins lösning (#3), jämfördes vid inkubering över natt i rumstemperatur samt 3-4 h i värmeskåp vid 56 ºC. Vävnader som färgades in var från paraffininbäddade klossar vilka tillhörde utsvarade fall med konstaterad hjärtinfarkt. Dessa snittades och värmdes fast på oladdade samt laddade objektglas. Efter färgning graderades infärgning efter kategorierna vävnadsdifferentiering och cellkomponenter med en poängskala från 0, ej bedömbar, till 3, optimal. Det protokoll med högst poäng optimerades för att hitta bästa inkuberingstid i PTAH-lösning, därefter implementerades protokollet genom att bekräfta korrekt infärgning vid upprepade infärgningar av hjärtmuskelvävnad. Protokoll #3 visade på högst poäng efter infärgning vid jämförelsen med protokoll #1 och #2 och under optimering framgick inkubering under 4 h i värmeskåp vid 56 ºC som mest gynnsam, då denna inkuberingstid även visade på mindre känslighet vid dehydrering. Slutsatsen blev att PTAH-färgning efter protokoll #3 med refixering i Bouins lösning med 4 h i värmeskåp gav bäst resultat, men färgnyansen varierar med tid mellan dödsfallet och obduktionen. / Acute myocardial infarction (MI) is a common cause of death globally. MI without symptoms or with atypical symptoms is usually first detected at clinical autopsy with histological analyses of the myocardium. Phosphtungstic acid haematoxylin (PTAH) is a method to visualize MI by staining myocytes, fibrin, contraction bands and nuclei blue while collagen stains reddish brown. The aim of this degree project is to optimize PTAH staining protocol for MI diagnosis. Three different staining protocols, two protocols with PTAH staining and Mallory Bleach solution and one protocol with PTAH staining with refixation in Bouin’s solution (#1-3), where compared along with incubation overnight in room temperature and at different hours in heat (56 ºC). Paraffin embedded tissues from myocardium from different autopsy cases diagnosed with MI were cut before mounting and heating on non-charged as well as charged slides. After staining, results were evaluated using scores 0-3 for two parameters; differences in tissues and cell components. Highest evaluated protocol where optimized to find the ultimate incubation in PTAH staining solution. For implementations, additional myocardial tissues from other cases were stained to confirm repeated results. Protocol #3 was evaluated higher in comparison with #1-2 and optimized to 4 h incubation in PTAH-solution in heating with staining results less sensitive to dehydration. In conclusion, staining protocol #3 with 4 h heating was optimal, however, vulnerable to prolonged morgue storage time losing the insensitivity of colour.

<p>Validerat; 20101217 (root)</p>

Gliomas are the most common primary tumors of the central nervous system believed to arise from glial cells. Invasive growth and inherent propensity for malignant progression make gliomas incurable despite extensive treatment. I have developed a life-like orthotopic glioma model and used this and other in vivo models to study basic mechanisms of glioma development and treatment. Previous studies had indicated that experimental gliomas could arise from glial stem cells and astrocytes. The present thesis describes the making and characterization of a novel mouse model, Ctv-a, where gliomas are induced from oligodendrocyte progenitor cells (OPCs). Our study shows that OPCs have the capacity to give rise to gliomas and suggests in light of previous data that the differentiation state of the cell of origin affects tumor malignancy. CDKN2A encodes p16INK4a and p14ARF (p19Arf in mouse) commonly inactivated in malignant glioma. Their roles in experimental glioma have been extensively studied and both proteins have tumor suppressor functions in glial stem cells and astrocytes. Here, we demonstrate that p19Arf only could suppress gliomagenesis in OPCs while p16Ink4a had no tumor suppressive effect. Functional DNA repair is pivotal for maintaining genome integrity, eliminating unsalvageable cells and inhibiting tumorigenesis. We have studied how RAD51, a central protein of homology-directed repair, affected experimental glioma development and have found that expression of RAD51 may protect against genomic instability and tumor development. Angiogenesis, the formation of new blood vessels from pre-existing ones, is a central feature of malignant progression in glioma. Antiangiogenic treatment by inhibition of vascular endothelial growth factor receptor signaling is used in the clinic for treatment of some cancers. We have investigated the effect of an alternative antiangiogenic protein, histidine-rich glycoprotein (HRG), on glioma development and found that HRG could inhibit the formation of malignant gliomas and completely prevent the formation of glioblastoma.

BACKGROUND: Prostate cancer is by far the most common cancer among Swedish men. Some patients have an aggressive lethal disease, but the majority of affected men have long expected survival. Unfortunately, the diagnostic tools available are insufficient in predicting disease aggressiveness. Novel prognostic markers are therefore urgently needed. Furthermore, metastatic prostate cancer is generally treated with castration, but the long-term effects are insufficient. Additional studies are therefore needed to explore how the effects of this therapy can be enhanced. Prostate growth and regression is beside testosterone controlled by locally produced regulators. Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) are two of the major regulators in the normal prostate and in prostate tumours. MATERIALS AND METHODS: VEGF and EGFR were explored in the prostate, by treating rats with either anti-VEGF or anti-EGFR treatment during castration and testosterone-stimulated prostate growth. Rats with implanted androgen-independent prostate tumours were treated with an inhibitor of both VEGF receptor-2 (VEGFR-2) and EGFR. Stereological techniques, immunohistochemistry, western blotting and quantitative real-time PCR were used to evaluate these experiments. Furthermore, prostate tissue from untreated prostate cancer patients was used to retrospectively explore the expression of phosphorylated-EGFR (pEGFR) in relation to outcome. RESULTS: Anti-VEGF treatment during testosterone-stimulated prostate growth, inhibited vascular and prostate growth. Anti-EGFR treatment during castration and testosterone-stimulated prostate growth resulted in enhanced castration effects and inhibited prostate growth. Anti-vascular treatment of androgen-independent prostate cancer with an inhibitor of VEGFR-2 and EGFR, that targets the normal and tumour vasculature, enhanced the effects of castration. Low immunoreactivity for pEGFR in prostate epithelial cells, both in the tumour and also in the surrounding non-malignant tissue, was associated with good prognosis. CONCLUSIONS: Anti-vascular treatment, with an inhibitor of VEGFR-2 and EGFR, in combination with castration could be an effective way to treat androgen-insensitive prostate tumours. VEGF and EGFR signalling are necessary components in testosterone-stimulated prostate growth. Phosphorylation of EGFR could be a useful prognostic marker for prostate cancer patients. Tumours may affect the surrounding non-malignant tissue and pEGFR immunoreactivity in the morphologically normal prostate tissue can be used to retrieve prognostic information.

Chronic lymphocytic leukemia (CLL) is a heterogeneous malignancy displaying varying clinical outcome, where molecular markers today can divide patients into prognostic subgroups. Despite the introduction of new agents for treatment, remissions are usually not sustained in CLL and resistance towards treatment can partly be explained by aberrant apoptosis. The aim of this thesis was to find new drugs for CLL patients resistant to conventional therapy and to analyze genes involved in apoptosis within different prognostic subgroups. In paper I-II, the in vitro activity of substances was investigated using the fluorometric microculture cytotoxicity assay (FMCA). When evaluating rapamycin (paper I), an inhibitor of mTOR, in 97 tumor samples from different entities, CLL was found to be one of the most sensitive tumor types. Combination experiments on patient CLL cells indicated that rapamycin acted synergistically with the CLL drugs vincristine and chlorambucil. An investigation of 20 anti-cancer agents in cells from 40 CLL patients (paper II) revealed that prednisolone and rolipram displayed high activity in poor-prognostic patients, in particular IGHV unmutated CLL. Furthermore, when used in combination these agents were found to produce a synergistic effect. In paper III, the anti-apoptotic BCL2 family member BFL1 was evaluated in 37 CLL cases. Levels of BFL1 were higher in fludarabine-resistant patients compared to fludarabine-sensitive patients. In addition, the high expression of BFL1 inversely correlated to fludarabine-induced apoptosis in CLL cells. A single nucleotide polymorphism in the anti-apoptotic BCL2 gene (-938C&gt;A) has been suggested as a novel poor-prognostic marker in CLL. In paper IV, we investigated this BCL2 polymorphism in 268 CLL patients and correlated genotypes to clinical data. However, no association could be confirmed between this polymorphism and clinical outcome or established prognostic markers. In conclusion, this thesis has shown that rapamycin is a potential drug for treatment in CLL. Furthermore, prednisolone and rolipram were identified as interesting candidates for treatment of poor-prognostic patients. Finally, the anti-apoptotic protein BFL1 may contribute to chemoresistance and hence represents a potential therapeutic target in CLL, whereas from our data, the BCL2 -938C&gt;A polymorphism does not appear to have any prognostic significance.

This thesis describes the development and use of a new class of molecular tools called Selector probes, and its potential for investigations of genetic variation. The Selector technology provides multiplex amplification of targeted DNA sequences with a high specificity, and an enrichment factor in the same order of magnitude as PCR. A common feature in this thesis work is to focus the analysis on DNA regions of interest. For example, this technique can be implemented in analysing candidate regions found by whole genome studies that need validation (global to local analysis), and applications requiring detection of rare alleles (common to rare allele), important in for example cancer samples. An assay is presented that allows for fast and simple quantification of relative copy-number variations. The method was proven to be able to detect aneuploidy in chromosome 13, 18, 21 and X, with a resolution enough to distinguish between 4 and 5 copies. The method was successfully applied to solve a biological question regarding a copy-number variation, that explains the Ridge phenotype typical for the dog bread Rhodesian Ridgebacks. The Selector strategy was able to detect and map a tandem duplication with a size of 133 kb, which was characterized with base-pair resolution. A readout platform that facilitates simultaneous digital quantitative analysis of a large numbers of biomolecules is further introduced. The work involves arraying amplified product from successful selection and decoding each molecule by hybridization of fluorophore labeled oligonucleotides. Finally, a genome partitioning method which is applied upstream of next generation sequencing platforms is presented. It is shown that the method provides successful enrichment with 98 % coverage and 94 % specificity and high enrichment uniformity. The technique was applied for mutation analysis of 26 cancer-related genes in tumor cell-lines and tissue.

Copy number variation (CNV) comprises a recently discovered kind of variation involving deletion and duplication of DNA segments of variable size, ranging from a few hundred basepairs to several million. By altering gene dosage levels or disrupting proximal or distant regulatory elements CNVs create human diversity. They represent also an important factor in human evolution and play a role in many disorders including cancer. Array-based comparative genomic hybridization as well as expression arrays are powerful and suitable methods for determination of copy number variations or gene expression changes in the human genome. In paper I we established a 32K clone-based genomic array, covering 99% of the current assembly of the human genome with high resolution and applied it in the profiling of 71 healthy individuals from three ethnic groups. Novel and previously reported CNVs, involving ~3.5% of the genome, were identified. Interestingly, 87% of the detected CNV regions overlapped with known genes indicating that they probably have phenotypic consequences. In papers II through IV we applied this platform to different tumor types, namely two collections of brain tumors, glioblastoma (paper II) and medulloblastoma (paper III), and a set of bladder carcinoma (paper IV) to identify chromosomal alterations at the level of DNA copy number that could be related to tumor initiation/progression. Tumors of the central nervous system represent a heterogeneous group of both benign and malignant neoplasms that affect both children and adults. Glioblastoma and medulloblastoma are two malignant forms. Glioblastoma often affects adults while the embryonal tumor medulloblastoma is the most common malignant brain tumor among children. The detailed profiling of 78 glioblastomas, allowed us to identify a complex pattern of aberrations including frequent and high copy number amplicons (detected in 79% of samples) as well as a number of homozygously deleted loci. These regions encompassed not only previously reported oncogenes and tumor suppressor genes but also numerous novel genes. In paper III, a subset of 26 medulloblastomas was analyzed using the same genomic array. We observed that alterations involving chromosome 17, especially isochromosome 17q, were the most common genomic aberrations in this tumor type, but copy number alterations involving other chromosomes: 1, 7 and 8 were also frequent. Focal amplifications, on chromosome 1 and 3, not previously described, were also detected. These loci may encompass novel genes involved in medulloblastoma development. In paper IV we examined for the presence of DNA copy number alterations and their effect on gene expression in a subset of 21 well-characterized Ta bladder carcinomas, selected for the presence or absence of recurrences. We identified a number of novel genes as well as a significant association between amplifications and high-grade and recurrent tumors which might be clinically useful. The results derived from these studies increase our understanding of the genetic alterations leading to the development of these tumor forms and point out candidate genes that may be used in future as targets for new diagnostic and therapeutic strategies.