Det här examensarbetet består i ett produktutvecklingsprojekt som utfördes i samarbete med Liquid Biopsy AB. Syftet med arbetet var att utveckla ett engångskassettsbaserat produktsystem baserat på företagets patentsökta metod för isolering av cancer celler i suspension, inklusive cirkulerande tumörceller. Liquid Biopsy AB är ett svenskt utvecklingsbolag som baserat på ny och unik teknik, är oberoende av proteinmarkörer, använder cirkulerande tumörceller och andra suspenderade tumörceller för att möjliggöra bättre och billigare cancerdiagnostik. Examensarbetet har fokuserat på utvecklingen av engångskassetten, men parallellt arbete har även utförts med tillhörande basenhet. Ulrich och Eppingers produktutvecklingsprocess har utgjort grunden för den process som följts i arbetet, dock med ökat fokus på testning och utvärdering. För att få en bredare kunskapsbas inleddes arbetet med en marknads- och omvärldsanalys samt informationsinsamling om utmaningar och medicintekniska krav. För att tydligt definiera produktvisionen utfördes även undersökningar med potentiella användarna, om företagets patentsökta metod och befintliga prototyper samt framtida förbättringspotential. Det kassettkoncept som utvecklats bygger på användning av provrör av existerande standard, få tillverkningsprocesser och god användarvänlighet, något som samtliga varit av hög prioritet under arbetet. För att säkerställa att produktens flödessystem fungerar som tänkt utfördes tester under prototypframtagningen. Testningen visade att konceptet fungerar i stort sett som tänkt med avseende på flöden, dock förekom vissa toleransproblem som följd av den valda prototypframtagningsprocessen, och vissa andra viktiga egenskaper återstår att testa. Resultatet av utvecklingsprocessen är en första fysisk prototyp av engångskassetten och en funktionell partiell prototyp av basenheten, motsvarande gränssnittet mot engångkassetten, för att möjliggöra testning av engångskassetten. Slutsatsen av arbetet är att det framtagna produktsystemet har tydliga fördelar gentemot företagets befintliga prototyper: inklusive att en engångskassett framtagits, att denna kan utgöra underlag för en produkt, och att denna bland annat har väsentligt kortare processväg vilken i sin tur borde kunna leda till förkortad processtid. Utförd finansiell analys visar även att framtaget produktsystem kan säljas till konkurrenskraftiga priser och med en betydligt lägre instegskostnad än dagens konkurrerande produkter. / This thesis consists of a product development project conducted in collaboration with Liquid Biopsy AB. The purpose of this work was to develop a disposable cartridge-based product system based on the company’s patent-pending method for isolation of circulating tumor cells and other suspended tumor cells. Liquid Biopsy AB is a Swedish medical technology research company with a unique new rheological technology, that is independent of protein markers, using suspended cancer cells, including circulating tumor cells, allows better and cheaper cancer diagnostics than today. The thesis work has focused on the development of the disposable cassette, but parallel work has also been performed with the associated base unit. Ulrich and Eppingers product development process has made up the basis for the process being followed in the thesis work, with increased focus on testing and evaluation. The work began with a market analysis and information gathering on challenges and medical requirements. Several activities were also carried out in order to clearly define the product vision, including user-surveys, analysis of the company's existing prototypes, as well as potential for future improvements. The developed cartridge concept is based on the use of standard test tubes, few manufacturing processes and user-friendliness which all have been high priorities in this work. The cartridge concept consists essentially of various plastic materials and is adapted for manufacturing by injection molding. To ensure that the product’s flow system was operating as intended, tests were conducted during the prototype phase. Testing showed that the concept design flows largely as intended, yet with some tolerance problems as a result of the selected rapid prototyping process, while other essential properties remain to be tested. The result of the development process is a first physical prototype of the disposable cartridge and a partial functional prototype of the base unit to allow testing with the disposable cartridge. The conclusion of this thesis work is that the developed product system has strong advantages over the company’s existing prototypes, including a first version of a disposable cassette that has potential to form the basis of a mass-producible product, significantly shorter processing route which in turn should allow a reduction of the processing time. Financial analysis also indicates that the designed product systems can be sold at competitive prices and with a significantly lower entry cost than today's rivaling products.

Orientador: Luiz Carlos Zeferino / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-12T08:55:33Z (GMT). No. of bitstreams: 1 Vieira_SabasCarlos_D.pdf: 1418388 bytes, checksum: 1d70cc451697cfbd93e48319a0284d4e (MD5) Previous issue date: 2008 / Resumo: Objetivos: Avaliar a detecção do linfonodo sentinela em pacientes com câncer do colo do útero utilizando a combinação de azul patente com tecnécio99m e complicações associadas ao uso do azul patente. Sujeitos e métodos: Este foi um estudo de uma série de casos, para o qual foram selecionadas 56 mulheres com diagnóstico de câncer do colo do útero estádios Ia2,Ib1,Ib2 e IIa da FIGO, que se submeteram ao procedimento de identificação do linfonodo sentinela. O período de realização do estudo foi de maio de 2006 a dezembro de 2007. O estudo é apresentado em dois artigos: o primeiro consiste na detecção do linfonodo sentinela no câncer do colo do útero pela combinação do azul patente com tecnécio 99m e avalia a concordância entre a linfocintigrafia pré-operatória e o mapeamento linfático intra-operatório com o gama probe; o segundo consiste na avaliação das alterações da oximetria de pulso das pacientes submetidas à cirurgia após a injeção do azul patente. Resultados: No primeiro artigo identificou-se pelo menos um linfonodo sentinela em 83,13% das pacientes e a localização mais freqüente destes linfonodos foi na cadeia ilíaca externa. A sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo foram, respectivamente, 80%, 100%, 100% e 97,67% no histopatológico de congelação. Além disso, observou-se que linfocintigrafia pré operatória detecta um número consideravelmente menor de linfonodos sentinelas quando comparado ao mapeamento linfático intra-operatório com o gama probe. No segundo estudo observou-se que somente uma paciente apresentou reação anafilática. Treze pacientes apresentaram queda de oximetria de pulso (menor que 96% de saturação) após a injeção do azul patente no colo do útero, que durou em média cinco minutos e sem repercussões clínicas; essa queda se associou de forma limítrofe com tumores maiores e localizados ao redor do orifício externo do canal cervical. Conclusões: Concluiu-se que a combinação do azul patente com o tecnécio99m demonstrou excelentes resultados na detecção do linfonodo sentinela; a linfocintigrafia pré-operatória não oferece qualquer vantagem em relação ao mapeamento linfático intra-operatório com azul patente e tecnécio99m. Não houve repercussões clínicas devido à queda da oximetria de pulso e essas alterações se correlacionaram, embora com significância limítrofe, com tumores maiores e localizados ao redor do orifício cervical externo. / Abstract: Objectives: To evaluate sentinel lymph node detection in cervical cancer patients using a combination of patent blue dye and technetium99m and assess complications associated with the use of patent blue dye. Subjects and methods: This study investigated a case series that selected 56 women diagnosed with FIGO stage Ia2, Ib1, Ib2 and IIa cervical cancer who underwent a procedure for sentinel lymph node identification. The study was conducted from May 2006 to December 2007 and was described in two articles. The first article was about a study of sentinel lymph node detection in cervical cancer using a combination of patent blue dye and technetium99m. It assessed the agreement between preoperative lymphoscintigraphy and intraoperative lymphatic mapping with a gamma probe. The second article focused on the evaluation of changes in pulse oximetry readings in patients undergoing surgery after patent blue injection. Results: In the first article, at least one sentinel lymph node was identified in 83.13% of the patients and the most frequent site for finding sentinel lymph nodes was the external iliac chain. The sensitivity, specificity, positive predictive value and negative predictive value were 80%, 100%, 100% and 97.67% respectively on histopathology examination of frozen biopsy. In addition, it was observed that preoperative lymphoscintigraphy detected a substantially lower number of sentinel lymph nodes when compared to intraoperative lymphatic mapping with a gamma probe. In the second study, only one patient presented with an anaphylactic reaction. Thirteen patients showed a decrease in pulse oximetry readings (less than 96% saturation) after patent blue injection into the cervix, which lasted an average of five minutes and had no clinical repercussions. There was a borderline association between this decline in oxygen saturation values and tumors that were larger and located around the external cervical os. Conclusions: It was concluded that a combination of patent blue dye and technetium99m demonstrated excellent results in the detection of sentinel lymph nodes. Preoperative lymphoscintigraphy offers no advantage in relation to intraoperative lymphatic mapping with patent blue dye and technetium99m. There were no clinical repercussions due to lower oxygen saturation values. These changes correlated with tumors that were larger and located around the external cervical os, although the significance of this correlation was borderline. / Doutorado / Ciencias Biomedicas / Doutor em Tocoginecologia

Objectives: We compared the transperineal MRI/ultrasoundfusion biopsy (fusPbx) to transrectal systematic biopsy (sys-Pbx) in patients with previously negative biopsy and investigated the prediction of tumour aggressiveness with regard to radical prostatectomy (RP) specimen. Material and Methods: A total of 710 patients underwent multiparametric magnetic resonance imaging (mpMRI), which was evaluated in accordance with Prostate Imaging Reporting and Data System (PI-RADS). The maximum PI-RADS (maxPI-RADS) was defined as the highest PI-RADS of all lesions detected in mpMRI. In case of proven prostate cancer (PCa) and performed RP, tumour grading of the biopsy specimen was compared to that of the RP. Significant PCa (csPCa) was defined according to Epstein criteria. Results: Overall, scPCa was detected in 40% of patients. The detection rate of scPCa was 33% for fusPbx and 25% for sysPbx alone (p < 0.005). Patients with a maxPI-RADS ≥3 and a prostate specific antigen (PSA)-density ≥0.2 ng/mL2 harboured more csPCa than those with a PSA-density < 0.2 ng/mL2 (41% [33/81] vs. 20% [48/248]; p < 0.001). Compared to the RP specimen (n = 140), the concordance of tumour grading was 48% (γ = 0.57), 36% (γ = 0.31) and 54% (γ = 0.6) in fusPbx, sysPbx and comPbx, respectively. Conclusions: The combination of fusPbx and sysPbx outperforms both biopsy modalities in patients with re-biopsy. Additionally, the PSA-density may represent a predictor for csPCa in patients with maxPI-RADS ≥3.

<p>The aims of this work were to image and analyze the needle behavior at automated core biopsy, to investigate the clinical utility of an alternative core biopsy technique using a semiautomated gun in breast and axillary lesions, and also to compare core biopsy with surgical specimens in malignant breast lesions regarding histologic features and hormone receptor expression.</p><p>In two experimental studies, using butter and silicon phantoms, respectively, the needle pass was imaged and its dynamic behavior studied. It was shown that the needle took a curved course in phantoms. It deviated to the same side as where the tip lay, and the degree of the curvature increased with increasing hardness of the phantoms. Our experimental methods can be applied for imaging of needle behavior and thereby improvement of needle configuration.</p><p>In two clinical studies, a semiautomated gun was used for large needle core biopsy of breast and axillary lesions in two series of 145 and 21 patients, respectively. The sensitivity of the method for diagnosis of malignancy was 87% (108/124), and in 37% (31/83) of cases the full length of the needle notch was filled with specimen. No injury to the neurovascular structures of the axillary area was observed. It was concluded that the semiautomated gun can be used as an alternative to the automated gun when the size and location of the lesion render use of the automatic device uncertain or dangerous, e.g., in small breast lesions or lesions located in the axilla.</p><p>In a series of 129 cases of breast cancer, comparison of core biopsy and surgical specimens showed that core biopsy provided enough information on the histologic type and grade of the lesions. Also, there was moderate to high concordance between the two methods for assessment of progesterone receptors and estrogen receptors (Spearman`s kappa 0.67 and 0.89, respectively).</p>

The aims of this work were to image and analyze the needle behavior at automated core biopsy, to investigate the clinical utility of an alternative core biopsy technique using a semiautomated gun in breast and axillary lesions, and also to compare core biopsy with surgical specimens in malignant breast lesions regarding histologic features and hormone receptor expression. In two experimental studies, using butter and silicon phantoms, respectively, the needle pass was imaged and its dynamic behavior studied. It was shown that the needle took a curved course in phantoms. It deviated to the same side as where the tip lay, and the degree of the curvature increased with increasing hardness of the phantoms. Our experimental methods can be applied for imaging of needle behavior and thereby improvement of needle configuration. In two clinical studies, a semiautomated gun was used for large needle core biopsy of breast and axillary lesions in two series of 145 and 21 patients, respectively. The sensitivity of the method for diagnosis of malignancy was 87% (108/124), and in 37% (31/83) of cases the full length of the needle notch was filled with specimen. No injury to the neurovascular structures of the axillary area was observed. It was concluded that the semiautomated gun can be used as an alternative to the automated gun when the size and location of the lesion render use of the automatic device uncertain or dangerous, e.g., in small breast lesions or lesions located in the axilla. In a series of 129 cases of breast cancer, comparison of core biopsy and surgical specimens showed that core biopsy provided enough information on the histologic type and grade of the lesions. Also, there was moderate to high concordance between the two methods for assessment of progesterone receptors and estrogen receptors (Spearman`s kappa 0.67 and 0.89, respectively).

Hintergrund und Zielsetzung: Jährlich erkranken in Westeuropa 4% der Gesamtbevölkerung an Akutem Nierenversagen (ANV). Zudem leiden 8,5% der Bevölkerung an einer Chronischen Niereninsuffizienz (CNI). Valide epidemiologische Daten über Erkrankungen der Niere existierten für Deutschland jedoch nicht, da zu diesem Zeitpunkt Erkrankungsfälle noch nicht flächendeckend systematisch erfasst wurden und das Deutsche Nierenregister noch im Aufbau war. Die vorliegende retrospektive Analyse untersucht und dokumentiert räumliche und zeitliche Veränderungen des histomorphologischen Befundspektrums aus Eigennierenbiopsien im Großraum Leipzig über den Zeitraum 1993-2012. Methoden: Als Grundlage der Arbeit dient ein zeitlich und geographisch strukturiertes Nierenbiopsieregister, welches für die vorliegende Arbeit aus den gesammelten histologischen Eigennierenbiopsiebefunden erstellt wurde. Zu jedem Fall wurde zur räumlichen Zuordnung, wenn ermittelbar, die Postleitzahl (PLZ) des Wohnortes des jeweiligen Patienten eingetragen. Bei einer annähernd vollständigen Erfassung der PLZ wurde für den jeweiligen Zeitraum für das Stadtgebiet Leipzig eine Inzidenzberechnung durchgeführt. Für das Leipziger Umland war dies nicht vorgesehen, da von einer unvollständigen Erfassung auszugehen war. Die statistische Auswertung erfolgte über Kontingenztafeln per Chi-Quadrat-Test oder per Varianzanalyse. Ergebnisse: In die Analyse des erstellten Biopsieregisters wurden n=943 Erstbiopsien eingeschlossen, unter vorherigem Ausschluss pädiatrischer Fälle und Folgebiopsien. Die IgA-Nephropathie (IgANP) war mit 19,5% die häufigste gestellte Diagnose, gefolgt von der Hypertensiven Nephropathie (HZNP) bzw. der Fokal-Segmentalen Glomerulosklerose (FSGS) und der Granulomatose mit Polyangiitis (GPA). Die räumliche Verteilung innerhalb der Untersuchungsregion unterlag teilweise großen Schwankungen. Die IgANP wurde im Leipziger Umland 36% häufiger beobachtet als im Leipziger Stadtgebiet. Auch im Zeitverlauf waren Schwankungen zu beobachten. Im Zeitraum 2009-2012 war die HTNP/FSGS mit 18.9% die häufigste Diagnose, gefolgt von der GPA mit 17,8% und der IgANP mit 15,6%. Zudem nahm die Häufigkeit der Glomerulopathie der dünnen Basalmembran (TBMD) bzw. des Alport-Syndroms stark ab. Auch die regionale Verteilung schwankte im Zeitverlauf stark. Auf Basis der ermittelten Postleitzahlen wurden für den Leipziger Stadtraum für den Zeitraum 2001-2009 jährliche Inzidenzen berechnet. Am Häufigsten trat dabei die GPA mit 0,9 (0,0-2,2) Fällen pro 100.000 Einwohner auf, gefolgt von der HTNP/FSGS mit 0,8 (0,2-2,2) und der IgANP mit 0,8 (0,2-1,4). Schlussfolgerung: Das Nephropathiespektrum im Großraum Leipzigs deckt sich, soweit konsolidierte Vergleichszahlen existieren, mit der bestehenden Literatur. In den Vergleichsstudien zeigte sich eine große Heterogenität. Einige Schwankungen, wie bei der HTNP/Alport-Syndrom oder bei der diabetischen Nephropathie (DNP) beobachtet, sind klar auf Variabilitäten in der Indikationsstellung zurückzuführen. Andere mögliche Einflussfaktoren wurden diskutiert.:1 Einleitung . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1 Sachstand . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1.1 Die Nierenbiopsie . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1.2 Äußere Einflussfaktoren auf Nierenerkrankungen . . . . . . . . 3 1.1.3 Das Nierenregister . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1.2 Fragestellung dieser Arbeit . . . . . . . . . . . . . . . . . . . . . . . . . 5 2 Patienten und Methoden . . . . . . . . . . . . . . . . . . . . . . . 6 2.1 Patienten . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2.1.1 Datenmaterial . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2.1.2 Datenerhebung . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2.2 Methoden . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 2.2.1 Datenstruktur . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 2.2.2 Statistische Auswertung . . . . . . . . . . . . . . . . . . . . . . . 11 2.2.3 Eingesetzte Software . . . . . . . . . . . . . . . . . . . . . . . . . 12 3 Ergebnisse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 3.1 Strukturierung der Patientendaten . . . . . . . . . . . . . . . . . . . . . 13 3.2 Regionale Verteilung der untersuchten Nierenbiopsiefälle . . . . . . . . 18 3.2.1 Regionale Verteilung der Glomerulopathien . . . . . . . . . . . 19 3.2.2 Regionale Verteilung nicht-glomerulärer Nephropathien . . . . 30 3.2.3 Zusammenfassung der Untersuchungsergebnisse über die regionale Verteilung . . . . . . . . . . . . . . . . . . . . . . . . . . 31 3.3 Zeitlicher Verlauf der untersuchten regionalen Diagnosenverteilung . 31 3.3.1 Zeitliche Dynamik der Glomerulopathien . . . . . . . . . . . . . 34 3.3.2 Zeitliche Dynamik der nicht-glomerulären Nephropathien . . . 48 3.3.3 Zusammenfassung der Untersuchungsergebnisse über die zeitliche Dynamik . . . . . . . . . . . . . . . . . . . . . . . . . . 51 4 Diskussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52 4.1 Beurteilung der Analyseergebnisse . . . . . . . . . . . . . . . . . . . . . 52 4.1.1 Aussagekraft der verwendeten Daten . . . . . . . . . . . . . . . 52 4.1.2 Evaluierung der Arbeitshypothesen . . . . . . . . . . . . . . . . 55 4.1.3 Ermittelte Inzidenzen . . . . . . . . . . . . . . . . . . . . . . . . 57 4.2 Analyse der Validität von Daten und Methoden . . . . . . . . . . . . . 60 4.2.1 Prozess der Datenerhebung . . . . . . . . . . . . . . . . . . . . . 60 4.2.2 Methoden der Datenanalyse . . . . . . . . . . . . . . . . . . . . 60 5 Ausblick . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 6 Zusammenfassung . . . . . . . . . . . . . . . . . . . . . . . . . . 65 7 Literaturverzeichnis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 A Anhang . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 / Background and Objectives: Annually, 4% of Western Europe\''s population fall ill with acute kidney injury (AKI). Furthermore, 8.5% of the same population are affected by chronic kidney disease (CKD). In Germany, valid nationwide epidemiological data on renal pathology didn\''t exist at the time of this study, although progress has been made with creating the German kidney biopsy register. This study analyzes temporal and spatial variances in the histomorphological spectrum of renal diseases of native kidney biopsies in the metropolitan area of Leipzig, Germany, from 1993 through 2012. Methods: For this study, a temporally and spatially structured kidney biopsy register was created from nephro-pathologic biopsy results. Spatial analysis was implemented by giving every entry its corresponding postal code. Unidentifiable entries were omitted. If the postal code was determined for every case within a timeframe, incidences for the city of Leipzig were calculated for the timeframe. Incidence for the surrounding areas were not calculated, because coverage was expected to be incomplete. Statistical analysis was done via Chi-Squared-Test or analysis of variances. Results: For this study n=943 cases were analyzed, omitting pediatric and follow-up biopsies. The leading diagnosis was IgA nephropathy (IgANP) with 19.5% (male: 22.1%, female: 15.4%), followed by hypertensive nephropathy (HTNP) resp. focal-segmental glomerulosclerosis (FSGS) and granulomatosis with polyangiitis (GPA). Spatial variance between the analyzed regions was high. Compared to the city of Leipzig, IgANP was observed one third more frequently in the surrounding regions. High temporal variance was also observed. From 2009 through 2012, HTNP/FSGS became leading diagnosis with 18.9%, followed by GPA with 17.8% and IgANP with 15.6%. Furthermore, frequency of thin base membrane disease (TBMD) resp. Alport\''s syndrome decreased sharply. Variance in spatial distribution was also observed over time. On the basis of determined postal codes, incidences for the city of Leipzig were calculated for the years 2001 through 2009. Highest annual incidence was observed in GPA with 0.9 (0.0-2.2) cases per 100 000 people, followed by HTNP/FSGS with 0.8 (0.2-2.2), IgANP with 0.8 (0.2-1.4). Conclusions: The spectrum of kidney pathology for the metropolitan area of Leipzig is in accordance with the data in literature, as far as consolidated figures were available. Results in compared studies were highly heterogenous. Some differences, e.g. decrease in TBMD resp. Alport\''s Syndrome or fluctuation of DNP, can be attributed to variance in indication for biopsy. External factors were discussed.:1 Einleitung . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1 Sachstand . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1.1 Die Nierenbiopsie . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1.2 Äußere Einflussfaktoren auf Nierenerkrankungen . . . . . . . . 3 1.1.3 Das Nierenregister . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1.2 Fragestellung dieser Arbeit . . . . . . . . . . . . . . . . . . . . . . . . . 5 2 Patienten und Methoden . . . . . . . . . . . . . . . . . . . . . . . 6 2.1 Patienten . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2.1.1 Datenmaterial . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2.1.2 Datenerhebung . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2.2 Methoden . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 2.2.1 Datenstruktur . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 2.2.2 Statistische Auswertung . . . . . . . . . . . . . . . . . . . . . . . 11 2.2.3 Eingesetzte Software . . . . . . . . . . . . . . . . . . . . . . . . . 12 3 Ergebnisse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 3.1 Strukturierung der Patientendaten . . . . . . . . . . . . . . . . . . . . . 13 3.2 Regionale Verteilung der untersuchten Nierenbiopsiefälle . . . . . . . . 18 3.2.1 Regionale Verteilung der Glomerulopathien . . . . . . . . . . . 19 3.2.2 Regionale Verteilung nicht-glomerulärer Nephropathien . . . . 30 3.2.3 Zusammenfassung der Untersuchungsergebnisse über die regionale Verteilung . . . . . . . . . . . . . . . . . . . . . . . . . . 31 3.3 Zeitlicher Verlauf der untersuchten regionalen Diagnosenverteilung . 31 3.3.1 Zeitliche Dynamik der Glomerulopathien . . . . . . . . . . . . . 34 3.3.2 Zeitliche Dynamik der nicht-glomerulären Nephropathien . . . 48 3.3.3 Zusammenfassung der Untersuchungsergebnisse über die zeitliche Dynamik . . . . . . . . . . . . . . . . . . . . . . . . . . 51 4 Diskussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52 4.1 Beurteilung der Analyseergebnisse . . . . . . . . . . . . . . . . . . . . . 52 4.1.1 Aussagekraft der verwendeten Daten . . . . . . . . . . . . . . . 52 4.1.2 Evaluierung der Arbeitshypothesen . . . . . . . . . . . . . . . . 55 4.1.3 Ermittelte Inzidenzen . . . . . . . . . . . . . . . . . . . . . . . . 57 4.2 Analyse der Validität von Daten und Methoden . . . . . . . . . . . . . 60 4.2.1 Prozess der Datenerhebung . . . . . . . . . . . . . . . . . . . . . 60 4.2.2 Methoden der Datenanalyse . . . . . . . . . . . . . . . . . . . . 60 5 Ausblick . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 6 Zusammenfassung . . . . . . . . . . . . . . . . . . . . . . . . . . 65 7 Literaturverzeichnis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 A Anhang . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73

<p>Breast cancer is the most common form of cancer in women in the United States. Histopathological examination through breast biopsy is considered as the "Gold Standard" for a definitive diagnosis. Contrast-enhanced Magnetic Resonance Imaging (MRI) is often used for guiding the biopsy in those cases in which the tumor may not be detectable under Ultrasound or X-ray mammography. Stabilization of the breast tissue during the biopsy is critical for its success to ensure that the target would not be displaced due to the patient movement or tissue deformation. Conventionally, the breast tissue is immobilized by firmly compressing it between two parallel plates. However, high compression forces causes significant patient discomfort and can reduce the intake of the contrast agent, which negatively impact the image quality.</p> <p>This thesis introduces devices and control methodologies for active tissue stabilization in magnetic resonance imaging (MRI)-guided breast biopsy. Pneumatic and piezoelectric actuators have been considered for developing concept designs for MRI-compatible tissue stabilization devices. Only the pneumatic device has been prototyped and tested. The device is comprised of two pneumatically-actuated support plates that would stabilize the biopsy target movements during needle insertion. An optimized geometry for the support plates allows for a good degree of tissue stabilization without relying on large compression forces. The plate configuration can also be adjusted inside the magnet bore using pneumatic actuators driven by pressure-controlled valves that are placed in the MR control room. This capability allows for the compensation of the target displacement based on MR image feedback. When combined with a separate needle drive mechanism, this stabilization device would enable in-bore MR-guided breast biopsy in combination with an in-bore needle driver system. The proposed approach offers improved target stabilization at reduced compression force and patient discomfort, that may also enhance MR image quality as result of greater intake of contrast agent. The open-front design of the stabilization plates provides greater flexibility in selecting the needle insertion entry point, and active adjustment of the support plates based on MR feedback improves the targeting accuracy.</p> <p>A concept design for a MR-compatible needle driver mechanism using piezoelectric actuators is also proposed. Experiments performed on chicken breast tissue with a prototype of the device demonstrate the effectiveness of this mechanism in increasing needle targeting accuracy using two simple error correction strategies. Furthermore, MRI compatibility tests are carried out to asses the performance of the device inside MRI.</p> / Master of Applied Science (MASc)

Thesis (PhD (Pathology. Anatomical Pathology))--University of Stellenbosch, 2009. / Dissertation presented for a PhD degree in anatomical pathology at Stellenbosch University. / ENGLISH ABSTRACT: Expediting a diagnosis of tuberculosis in children, particularly those who are immunocompromised due to HIV/AIDS, is essential, as they are vulnerable to develop severe forms of disease due to their immature or compromised immune systems. A significant percentage of children (8 to 10%) with TB have TB lymphadenitis, in isolation, or in combination with other disease manifestations. Fine needle aspiration biopsy (FNAB) is a simple and minimally invasive procedure well tolerated by children. It may be performed as an outpatient procedure by clinicians as well as nurses, and excellent results can be achieved with training in the correct procedure. The aim of this dissertation was to demonstrate that FNAB may contribute significantly to the diagnosis of mycobacterial lymphadenitis, with particular reference to children TB suspects. We first established that TB lymphadenitis is a common clinical problem in children in TB endemic areas and that FNAB is an efficient simple and effective diagnostic modality in children with peripheral lymphadenopathy. We then proceeded to document the diagnostic yield and time to diagnosis of FNAB compared to conventional laboratory specimens collected in children. We investigated the value of additional diagnostic modalities such as autofluorescence in improving the ability of cytology to make a definitive diagnosis of mycobacterial infection based on cytomorphology and identification of the organism. In countries where organisms such as Mycobacterium bovis BCG and nontuberculous mycobacteria are prevalent, culture with subsequent speciation is essential. The amount of material harvested during FNAB is minuscule, and requires immediate bedside inoculation for optimal yields. We developed an inexpensive and effective transport medium to facilitate mycobacterial culture from FNAB, even if this is collected at an outside facility. It is ideally suited for use in clinics and rural hospitals as it is stable at room temperature, maintains viability of the organism for seven days, and the closed lid format reduces contamination. Mycobacterial culture even using liquid-based media, takes up to 6 weeks, and this delay is unacceptable particularly in children. We developed a Nucleic Acid Amplification Technique (NAAT) using High Resolution Melt Analysis and applied this novel technique to FNAB specimens submitted in transport medium. Although sensitivity remained suboptimal, the technique is highly specific, simple and rapid. Its use could be incorporated into routine microbiology laboratories, to assist with rapid diagnosis while cultures are pending. We collected a solid body of evidence, which will promote the use of FNAB in suspected mycobacterial lymphadenopathy, particularly in children in resource-limited countries. The utilisation of the diagnostic methods identified will expedite speciation and allow early and appropriate initiation of therapy. This is in keeping with Millennium Development Goal 6: to combat TB by early detection of new cases and effective treatment. / AFRIKAANSE OPSOMMING: Kinders met tuberkulose (TB), en veral diegene met gekompromiteerde immuniteit as gevolg van MIV/VIGS, het ‘n verhoogde neiging om ernstige siektebeelde te ontwikkel vanweë hul onvolwasse of gekompromiteerde immuunsisteme. ‘n Spoedige diagnose van TB in kinders is dus noodsaaklik. ‘n Betekenisvolle persentasie van kinders (8 tot 10%) met TB het TB limfadenitis met of sonder meegaande ander siekteverskynsels. Fynnaaldaspirasiebiopsie (FNAB) is ‘n eenvoudige en minimale indringende prosedure wat geredelik deur kinders aanvaar word. Geneeshere en verpleegkundiges wie toepaslike opleiding in die uitvoering van FNAB ontvang het, kan die prosedure op buitepasiënte uitvoer en uitstekende resultate behaal. Die doel van hierdie studie was om aan te toon dat FNAB betekenisvol kan bydra tot die diagnose van mikobakteriële limfadenitis in veral kinders met vermoedelike TB. Daar was eerstens bevestig dat TB limfadenitis ‘n algemene kliniese probleem is in kinders in TB endemiese areas en dat FNAB ‘n doeltreffende, eenvoudige en effektiewe diagnostiese modaliteit is in kinders met perifere limfadenopatie. Vervolgens was FNAB se diagnostiese opbrengs en die tydsverloop tot diagnose vergelyk met dié van konvensionele laboratoriummonsters wat in kinders verkry word. Die bydrae van verdere diagnostiese modaliteite soos outofluoressensie tot ‘n verbetering in sitologie se rol in die diagnose van mikobakteriële infeksie, soos gebaseer op sitomorfologie en identifisering van organismes, is ondersoek. In lande waar organismes soos Mycobacterium bovis BCG en nie-tuberkuleuse mikobakterië heersend is, is kultuur en spesiebepaling noodsaaklik. Die hoeveelheid materiaal wat met FNAB verkry word is baie min en vereis onmiddellike okulasie vir die beste resultate. Tydens hierdie studie is ‘n goedkoop en effektiewe vervoermedium ontwikkel om mikobakteriële kultuur van FNAB verkreë monsters te fasiliteer, selfs al is die monster vanaf ‘n buite fasiliteit bekom. Die vervoermedium is baie geskik vir gebruik in klinieke en plattelandse hospitale. Dit is stabiel by kamertemperatuur, handhaaf lewensvatbaarheid van organismes vir sewe dae, en die geslote dekselformaat verminder kontaminasie. Mikobakteriële kultuur neem tot ses weke, selfs met die gebruik van vloeistofgebaseerde mediums. Sodanige vertraging in die diagnose is veral in kinders onaanvaarbaar. Tydens hierdie studie is ‘n Nukleïnsuur Amplifikasietegniek ontwikkel deur die aanwending van Hoë Resolusie Smeltanalise en is hierdie nuwe tegniek toegepas op FNAB verkreë monsters wat in die vermelde vervoermedium versamel was. Alhoewel sensitiwiteit nie optimaal was nie, is die tegniek baie spesifiek, eenvoudig en vinnig. Dit kan in roetine mikrobiologie laboratoriums gebruik word om vinnige diagnose te bewerkstellig terwyl daar gewag word vir die kultuur se resultaat. Hierdie studie bied omvattende bewys ter ondersteuning van die gebruik van FNAB in veral kinders met vermoedelike mikobakteriële limfadenopatie in lande met beperkte hulpbronne. Die toepassing van die diagnostiese metodes wat in hierdie studie identifiseer is sal spesiebepaling bespoedig en vroegtydige en toepaslike behandeling verseker. Dit stem ooreen met Millennium Ontwikkelingsdoelwit 6: om TB te beveg deur vroeë opsporing van nuwe gevalle en effektiewe behandeling.

Background: Minor procedures in surgery and urology such as groin hernia and hydrocele repair, as well as prostate biopsies are very frequently done in routine practice. Complications and insufficient outcomes thus affecting many patients and the cumulative effect of this are of major importance in a population perspective. Aim: To explore complications and outcomes of surgical or diagnostic procedures and possible risk factors or predictors for adverse effects. Methods: By using both national quality and administrative registers, and by complementing registers with patient reported outcome measures, examine outcomes such as complications, persistent pain and recurrences. Also, in the case of hydro and spermatoceles, report incidence numbers. Further, by using a randomized trial, explore minimally invasive procedure such as sclerotheraphy compared to conventional surgery in respect to cure and adverse events. Results: When comparing with the open anterior mesh repair, endoscopic technique is advantageous in respect to the patient reported outcome of persistent pain. The drawback was an increased risk of postoperative complications and reoperation for recurrence. Incidence numbers for hydro and spematocele were 100/100000 men. Aspiration (± sclerotherapy) had a significantly lower rate of complications as compared to conventional surgery. In the interim analysis of the randomized trial, comparing sclerotherapy to Lord´s procedure for hydroceles, the cure rate was similar between treatments. Definite conclusions cannot be made due to the risk of type 2 errors, and the study will thus continue. In the case of trans-rectal prostate biopsy, the rates increased every year during the study time frame, up to an approximate risk of two per cent in 2012 for hospital readmission within 30 days, without an increased mortality within 30 days. Conclusions: The open anterior mesh procedure is still the preferred method for groin hernia repair in routine surgical practice. Hydro and spermatocele surgery is associated with high rates of complications, and the indication for repair should be scrutinized. The rates of infection after prostate biopsy is increasing and methods to reduce unnecessary biopsies as well as improved prophylaxis should be investigated.

Doctor of Philosophy / Department of Chemistry / Stefan H. Bossmann / Cancer is a group of diseases that are characterized by uncontrolled growth and spread of cells. In order to treat cancer successfully, it is important to diagnose cancers in their early stages, because survival often depends on the stage of cancer detection. For that purpose, highly sensitive and selective methods must be developed, taking advantage of suitable biomarkers. The expression levels of proteases differ from one cancer type to the other, because different cancers arise from different cell types. According to the literature, there are significant differences between the protease expression levels of cancer patients and healthy people, because solid tumors rely on proteases for survival, angiogenesis and metastasis. Development of fluorescence-based nanobiosensors for the early detection of pancreatic cancer and non-small-cell lung cancer is discussed in this thesis. The nanobiosensors are capable of detecting protease/arginase activities in serum samples over a broad range. The functionality of the nanobiosensor is based on Förster resonance energy transfer and surface energy transfer mechanisms. The nanobiosensors for protease detection feature dopamine-coated Fe/Fe₃O₄ nanoparticles, consensus (cleavage) peptide sequences, meso-tetra(4-carboxyphenyl)porphine (TCPP), and cyanine 5.5. The consensus peptide sequences were synthesized by solid-supported peptide synthesis. In this thesis, improved consensus sequences were used, which permit faster synthesis and higher signal intensities. TCPP, which is the fluorophore of the nanoplatform, was connected to the N-terminal end of the oligopeptides while it was still on the resin. After the addition of TCPP, the TCPP-oligopeptide was cleaved off the resin and linked to the primary amine groups of Fe/Fe₃O₄-bound via a stable amide bond. In the presence of a particular protease, the consensus sequences attached to the nanoparticle can be cleaved and release TCPP to the aqueous medium. Upon releasing the dye, the emission intensity increases significantly and can be detected by fluorescence spectroscopy or, similarly, by using a fluorescence plate reader. In sensing of arginase, posttranslational modification of the peptide sequence will occur, transforming arginine to ornithine. This changes the conformational dynamics of the oligopeptide tether, leading to the increase of the TCPP signal. This is a highly selective technology, which has a very low limit of detection (LOD) of 1 x 10⁻¹⁶ molL⁻¹ for proteases and arginase. The potential of this nanobiosensor technology to detect early pancreatic and lung cancer was demonstrated by using serum samples, which were collected from patients who have been diagnosed with pancreatic cancer and non-small cell lung cancer at the South Eastern Nebraska Cancer Center (lung cancer) and the University of Kansas Cancer Center (pancreatic cancer). As controls, serum samples collected from healthy volunteers were analyzed. In pancreatic cancer detection, the protease/arginase signature for the detection of pancreatic adenocarcinomas in serum was identified. It comprises arginase, MMPs -1, - 3, and -9, cathepsins -B and -E, urokinase plasminogen activator, and neutrophil elastase. For lung cancer detection, the specificity and sensitivity of the nanobiosensors permit the accurate measurements of the activities of nine signature proteases in serum samples. Cathepsin -L and MMPs-1, -3, and -7 permit detecting non-small-cell lung-cancer at stage 1.