In 2012, more than 1.6 million new cases of breast cancer were diagnosed and about half a million women died of breast cancer. The incidence has increased in the developing world. The mortality, however, has decreased. This is thought to partly be the result of advances in diagnosis and treatment. Studying tissue samples from biopsies through a microscope is an important part of diagnosing breast cancer. Recent techniques include camera-equipped microscopes and whole slide scanning systems that allow for digital high-throughput scanning of tissue samples. The introduction of digital pathology has simplified parts of the analysis, but manual interpretation of tissue slides is still labor intensive and costly, and involves the risk for human errors and inconsistency. Digital image analysis has been proposed as an alternative approach that can assist the pathologist in making an accurate diagnosis by providing additional automatic, fast and reproducible analyses. This thesis addresses the automation of conventional analyses of tissue, stained for biomarkers specific for the diagnosis of breast cancer, with the purpose of complementing the role of the pathologist. In order to quantify biomarker expression, extraction and classification of sub-cellular structures are needed. This thesis presents a method that allows for robust and fast segmentation of cell nuclei meeting the need for methods that are accurate despite large biological variations and variations in staining. The method is inspired by sparse coding and is based on dictionaries of local image patches. It is implemented in a tool for quantifying biomarker expression of various sub-cellular structures in whole slide images. Also presented are two methods for classifying the sub-cellular localization of staining patterns, in an attempt to automate the validation of antibody specificity, an important task within the process of antibody generation.  In addition, this thesis explores methods for evaluation of multimodal data. Algorithms for registering consecutive tissue sections stained for different biomarkers are evaluated, both in terms of registration accuracy and deformation of local structures. A novel region-growing segmentation method for multimodal data is also presented. In conclusion, this thesis presents computerized image analysis methods and tools of potential value for digital pathology applications.

The objectives of this study was to examine the efficacy and safety of graded doses of c9, t11, t10, c12 CLA isomers on body composition, energy expenditure, lipid profile and hepatic biomarkers in hamsters. Male Golden Syrian hamsters (n=105) were randomized to seven treatments (control; 1, 2, 3% of c9, t11; 1, 2, 3% of t10, c12) for 28 days. Compared with control, 1% and 3% t10, c12 had lowered food intake with all three doses of t10, c12 lowering (p<0.0001) body fat mass (g). Groups fed with 1, 2, 3% t10, c12 and 3% c9, t11 treatments showed higher lean mass compared to control and other treatment groups. However, neither body weights, nor serum HDL or triglyceride levels differed across treatment groups. The 3% t10, c12 groups exhibited higher (p<0.0001) cholesterol and LDL-C levels compared to control or other treatment groups. The 2% and 3% t10, c12 groups also presented elevated ALT level (p<0.05). The present data suggest that 3% t10, c12 possess potential adverse effects on liver and posing unfavorable change in lipid profile.

Statement of Problem Telomere dynamics in the three-dimensional (3D) space of the mammalian nucleus plays an important role in the maintenance of genomic stability. However, the telomere distribution in 3D nuclear space of normal and tumor cells was unknown when the study was initiated. Methods Telomere fluorescence in situ hybridization (FISH) and 3D molecular imaging, deconvolution, and analysis were used to investigate telomere organization in normal, immortalized and tumor cells from mouse and human cell lines, and primary tissues. Results Telomeres are organized in a non-overlapping manner and in a cell-cycle dependant fashion in normal cells. In the late G2 phase of cell cycle, telomeres are assembled into a flattened sphere that is termed the telomeric disk In contrast, the telomeric disk is disrupted in the tumor cells. Moreover, telomeric aggregates (TAs) are found in tumor cells. Conditional c-Myc over-expression induces telomeric aggregation leading to the onset of breakage-bridge-fusion cycles and subsequent chromosomal abnormality. Conclusions Telomeres are distributed in a nonrandom and dynamic fashion in the 3D space of a normal cell. Telomeric aggregates are present in cells with genomic instability such as tumor cells and cells with deregulation of c-Myc. Consequently, TA can be a useful biomarker for research in cancer and other disease processes.

No description available.

Biomarkers measured in the blood can provide information about disease pathophysiology, diagnosis and prognosis. Pronounced proinflammatory responses are characteristic of severe malaria, and excessive activation of the immune system is central to the pathophysiology of both cerebral malaria and placental malaria. Severe malaria is characterized by cytoadherence of parasitized erythrocytes to the microvasculature; impaired tissue perfusion; dysregulated inflammatory responses; and activation of the complement system, mononuclear cells, and endothelium. Despite the availability of effective antimalarial drugs, the mortality rate in severe malaria remains unacceptably high. To glean further insight into malaria pathophysiology, we investigated host biomarkers of immune activation in the blood of subjects with different manifestations of severe disease. C5 has been identified as being necessary and sufficient for the development of experimental cerebral malaria. We hypothesized that C5a (a terminal component of the complement cascade with potent inflammatory properties) may mediate its action by inducing and exacerbating inflammatory processes in severe malaria, leading to endothelial activation and dysregulated angiogenesis. I tested this hypothesis in vitro, and found that C5a interacted with malaria toxin PfGPI to drive deleterious inflammatory and anti-angiogenic responses. As C5a and anti-angiogenic factor sFlt-1 have been implicated in models of pathologic pregnancies, we asked whether increased levels of C5a in subjects with placental malaria were associated with altered angiogenesis and poor birth outcomes. Our results suggest that C5a impairs angiogenic remodelling in placental malaria leading to vascular insufficiency and fetal growth restriction. Further, altered profiles of inflammatory and angiogenic biomarkers in the periphery may identify occult placental malaria infections. We extended these observations to cerebral malaria where similar pathogenic pathways contribute to disease pathophysiology. In adults and children with cerebral malaria, altered profiles of angiogenic proteins were associated with disease severity and mortality and represent putative diagnostic and prognostic biomarkers in severe malaria.

Biomarkers measured in the blood can provide information about disease pathophysiology, diagnosis and prognosis. Pronounced proinflammatory responses are characteristic of severe malaria, and excessive activation of the immune system is central to the pathophysiology of both cerebral malaria and placental malaria. Severe malaria is characterized by cytoadherence of parasitized erythrocytes to the microvasculature; impaired tissue perfusion; dysregulated inflammatory responses; and activation of the complement system, mononuclear cells, and endothelium. Despite the availability of effective antimalarial drugs, the mortality rate in severe malaria remains unacceptably high. To glean further insight into malaria pathophysiology, we investigated host biomarkers of immune activation in the blood of subjects with different manifestations of severe disease. C5 has been identified as being necessary and sufficient for the development of experimental cerebral malaria. We hypothesized that C5a (a terminal component of the complement cascade with potent inflammatory properties) may mediate its action by inducing and exacerbating inflammatory processes in severe malaria, leading to endothelial activation and dysregulated angiogenesis. I tested this hypothesis in vitro, and found that C5a interacted with malaria toxin PfGPI to drive deleterious inflammatory and anti-angiogenic responses. As C5a and anti-angiogenic factor sFlt-1 have been implicated in models of pathologic pregnancies, we asked whether increased levels of C5a in subjects with placental malaria were associated with altered angiogenesis and poor birth outcomes. Our results suggest that C5a impairs angiogenic remodelling in placental malaria leading to vascular insufficiency and fetal growth restriction. Further, altered profiles of inflammatory and angiogenic biomarkers in the periphery may identify occult placental malaria infections. We extended these observations to cerebral malaria where similar pathogenic pathways contribute to disease pathophysiology. In adults and children with cerebral malaria, altered profiles of angiogenic proteins were associated with disease severity and mortality and represent putative diagnostic and prognostic biomarkers in severe malaria.

Herzinsuffizienz ist eine sehr häufige Erkrankung im hohen Lebensalter mit zudem signifikant hoher Mortalität - vergleichbar mit der Mortalität häufiger Krebsarten. Biomarker wie die natriuretischen Peptide sind von großer Wichtigkeit hinsichtlich der Diagnosestellung und Prognoseabschätzung. Auch inflammatorische Marker, Copeptin sowie Mid-regionales Adrenomedullin (MR-proADM) haben eine wichtige Rolle sowohl in der Diagnosestellung der Herzinsuffizienz als auch in der Prognoseabschätzung eingenommen. Die Aussagekraft der Biomarker in einem diagnostisch naiven Kollektiv mit dem klinisch-anamnestischen Verdacht auf das Vorliegen einer Herzinsuffizienz ist jedoch bisher kaum untersucht worden. Die Handheld-BNP-Studie schloss diagnostisch naive Patienten ein, die sich mit Symptomen passend zu einer Herzinsuffizienz beim Hausarzt vorstellten. Binnen 14 Tagen erfolgte die Referenzdiagnose durch einen niedergelassenen Kardiologen. Ziel war es, die diagnostische Aussagekraft von BNP und der miniaturisierten Echokardiographie im primärärztlichen Bereich zu überprüfen. Die vorliegenden Follow-Up-II-Untersuchung untersuchte die prognostische Aussagekraft moderner Biomarker (N-terminales B-natriuretisches Peptid (NT-proBNP), Mid-regionales atriales natriuretisches Peptid (MR-proANP), Mid-regionales Adrenomedullin (MR-proADM), Copeptin, Tumornekrosefaktor Alpha (TNF- α) und hochsensitives C-reaktives Protein (hsCRP)). Die Endpunkte waren Tod jeder Ursache sowie kardiovaskulärer Tod. Insgesamt traten in unseren Analysen die natriuretischen Peptide mit ihrer prognostischen Aussagekraft hervor. In den univariaten Analysen zeigte sich das NT-proBNP als wichtigster Biomarker und in den multivariaten Analysen das MR-proANP. Bei diagnostisch naiven Patienten, die sich mit Herzinsuffizienzsymptomen bei ihrem Hausarzt vorstellen, besteht ein hohes Mortalitätsrisiko. Um diese Patienten adäquat zu selektieren, eine leitliniengerechte Therapie einzuleiten und um das Fortschreiten der Erkrankung aufzuhalten, ist eine frühzeitige Diagnosestellung beim Kardiologen wichtig. Natriuretische Peptide sind prädiktiv, jedoch stellt das MR-proANP aufgrund fehlender generalisierter Verfügbarkeit keine realistische Option im primärärztlichen Bereich dar. Das NT-proBNP hat eine flächendeckende Verfügbarkeit und wird mittlerweile in den Herzinsuffizienz-Leitlinien der ESC bei der Verdachtsdiagnose Herzinsuffizienz standardmäßig empfohlen. / Heart failure (HF) is a common burden in elderly patients and is associated with a significantly higher mortality - comparable to the mortality of common cancers. Natriuretic peptides are well-established and easily obtainable biomarkers serving in both daily clinical diagnostics and long-term prognostic evaluation in patients with HF. Furthermore, inflammatory markers, copeptin and mid-regional pro-adrenomedulline (MR-proADM) play a key role in the diagnostic and prognostic assessment of HF. However, little is known about the prognostic significance of modern cardiac biomarkers in diagnostically naive patients who present to their general practitioner (GP) with symptoms and signs potentially indicative of HF. The Handheld-BNP study evaluated diagnostically naive patients who attended their GP with suspected HF. Within 14 days, reference diagnosis was made by a cardiologist. The study examined the diagnostic and prognostic value of B-type NP measurements and the use of portable echocardiography in primary care. The Follow-Up-Study II investigated the prognostic relevance of modern cardiac biomarkers [N-terminal B-natriuretic peptide (NT-proBNP), mid-regional pro-atrial natriuretic peptide (MR-proANP), mid-regional pro-adrenomedullin (MR-proADM), copeptin, tumor necrosis factor alpha (TNF- α) and highly sensitive C-reactive protein (hsCRP)]. Endpoint analysis focused on all-cause mortality and cardiovascular mortality. Overall, natriuretic peptides showed the highest prognostic power (NT-proBNP in univariate and MR-proANP in multivariate analysis) and are therefore valuable tools in daily clinical care of HF-naive patients presenting with HF-associated symptoms to their GP.

Zugl.: Berlin, Humboldt-Univ., Diss., 2008

Zugl.: Berlin, Freie Universiẗat, Diss., 2008.

Regensburg, University, Diss., 2005.