京都大学 / 0048 / 新制・課程博士 / 博士(人間健康科学) / 甲第20295号 / 人健博第43号 / 新制||人健||4(附属図書館) / 京都大学大学院医学研究科人間健康科学系専攻 / (主査)教授 高桑 徹也, 教授 藤井 康友, 教授 武藤 学 / 学位規則第4条第1項該当 / Doctor of Human Health Sciences / Kyoto University / DGAM

A dissertation submitted to the Faculty of Health Sciences, University of Witwatersrand, in fulfilment of the requirements for the degree of Master of Science (Medicine) in the branch of Anatomical Pathology Johannesburg, 2012 / Objective To determine the effectiveness of the cell block technique for immunocytochemical diagnosis by comparing cytomorphologic preservation and immunocytochemistry (ICC) stains in paired cell block and conventional fine needle aspiration (FNA) samples. Study Design This was a prospective study. Material for both conventional smears and cell blocks were collected simultaneously during fine needle aspiration of 50 lesions comprising lymph node, lung and liver masses. Grading of cellularity, morphological preservation, architectural preservation, immunocytochemical staining intensity and presence of background staining were compared on paired FNA smears and cell block samples derived from the same case. Each arm of the paired analysis was performed blindly without knowledge of the grading outcome of the other. The Kappa statistic (Κ) was used to measure inter-rater agreement. Results The fifty samples evaluated included FNAs from the lung, 24/50 (48%); liver, 23/50 (46%) and lymph node, 3/50 (6%). The immunocytochemistry stains consisted of 44/50 (88%) CK7, 44/50 (88%) CK20, 18/50 (36%) TTF1, 10/50 (20%) synaptophysin, 10/50 (20%) Hepar-1 and 7/50 (14%) AE1/3. There was no overall agreement in preservation of cytomorphological detail and ICC staining between the two methods. The Papanicolaou stained conventional FNA smears fared better then cell block for the vi evaluation of nuclear and cytomorphologic characteristics; cells in the cell block were poorly preserved in many cases. The ICC stains worked better on the cell block samples due to lack of background and aberrant staining. Conclusion Conventional FNA smears and cell blocks complement each other. Our results indicate that it would be optimal to use both modalities in the diagnostic work-up of mass lesions amenable to FNA diagnosis; the former to assess morphology, and the latter for optimal immunocytochemistry results. In resource constrained settings, the cost implications of performing both conventional and blocked smears on all FNA material warrants further evaluation.

Orientador: Noeme Sousa Rocha / Resumo: O Osteossarcoma (OSA) refere-se ao tumor ósseo mais comum em cães. A citoinclusão compreende o ato de inserir amostras citológicas em blocos de parafina, auxiliando na qualidade do diagnóstico citológico e obtendo espécimes de reserva para serem utilizadas em outras técnicas, incluindo imunoistoquímica. Com o intuito de avaliar a sensibilidade, especificidade e descrição da técnica de citoinclusão no diagnóstico de OSA de ossos longos de cães, 11 cães de oito a 13 anos com diagnóstico de OSA foram submetidos a exames citológicos, histopatológicos e, técnica da citoinclusão. Para a citoinclusão, foi feita a punção do tumor, fixação com 1mL de álcool a 95%, deixado em repouso por cinco minutos na própria seringa e, aspirou-se aproximadamente 9 mL de formol a 10% e manteve-se fixado por mais 24 horas, mantendo a seringa posicionada para cima para decantação das células. Como imunomarcador, utilizou-se a osteopontina. Em razão da maior preservação arquitetural, a avaliação a partir das amostras de citoinclusão, verificou-se parte de relações arquiteturais mais bem estabelecidas do que em amostras de esfregaços, auxiliando na caracterização do processo. Houve concordância da citoinclusão com a histopatologia e marcação imunoistoquímica de osteopontina positiva com alta sensibilidade e especificidade. É possível afirmar, que a técnica de citoinclusão é eficaz, segura para pesquisa de marcadores prognósticos de tumores ósseos em menor espaço de tempo e baixo custo podendo ser utiliz... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

Aramide and arylene ether multiblock copolymers of (AB)n-type with various degrees of sulfonation have been prepared for use in direct methanol fuel cells. / Aramid- und Arylethersulfon-Multiblockcopolymere vom Typ (AB)n mit unterschiedlichem Sulfonierungsgrad wurden hergestellt und hinsichtlich ihrer Eignung als Polymerelektrolyte in der Direkt-Methanol-Brennstoffzelle getestet.

Fullerenhaltige Bulk-Heterojunction-Solarzellen auf Polymerbasis zeigen derzeit eine geringe Langzeitstabilität, die unter anderem auf der Entmischung der Bulkphasen beruht. In dieser Arbeit wurden daher auf neuartige Weise Blockcopolymere entwickelt, die zur Stabilisierung der Phasen dienen können. Ausgehend von Poly-3-hexylthiophen-Makroinitiatoren wurde ein zweiter Block mit reaktivem Comonomer polymerisiert, das zur Anbindung von reinem Fullerens bzw. von Fullerenderivaten diente. Die fullerenfunktionalisierten Polymere wurden in Modell-Systeme eingemischt und zeigten erste Verbesserungen bezüglich der Phasenstabilisierung.:1 Einleitung und Zielstellung 2 Grundlagen 2.1 Polymere Solarzellen 2.1.1 Theoretische Grundlagen und Funktionsweise 2.1.2 Materialien und Materialoptimierung 2.1.3 Stabilisierung der Blendmorphologien 2.2 Blockcopolymere – Eigenschaften und Aufbau 2.2.1 Kontrolliert radikalische Polymerisationen 2.2.2 Kumada-Catalyst Transfer Polycondensation 2.3 Fullerene – Eigenschaften und Funktionalisierung 3 Ergebnisse und Diskussion 3.1 Darstellung von Akzeptor-Polymeren 3.1.1 Synthese der Copolymere 3.1.2 Bingel-Reaktion 3.1.3 Polymeranaloge Reaktionen und Anbindung von Fullerenen an Copolymere 3.2 Synthesen von Donor-Akzeptor-Blockcopolymeren 3.2.1 Synthese und Charakterisierung des Makroinitiators und der Blockcopolymere 3.2.2 Polymeranaloge Reaktionen an Blockcopolymeren 3.3 Charakterisierung der Eigenschaften von Donor-Akzeptor-Blockcopolymeren in Blends 3.3.1 Blends mit fullerenhaltigen Blockcopolymeren 3.3.2 Blends mit azidfunktionalisiertem Blockcopolymer 3.3.3 Solarzellen-Tests 4 Zusammenfassung und Ausblick 5 Experimenteller Teil 5.1 Verwendete Chemikalien und Reagenzien 5.2 Geräte und Hilfsmittel 5.3 Synthesen niedermolekularer Verbindungen 5.4 Polymersynthesen 5.5 Polymeranaloge Reaktionen 6 Literaturverzeichnis Anhang

Malignt mesoteliom är en ovanlig och aggressiv cancerform. Den är vanligast i pleura och kallas då pleuralt mesoteliom. Vanligaste orsaken till utveckling av pleuralt mesoteliom är exponering för asbest. Insjuknande patienter har dålig prognos och begränsade behandlingsmöjligheter. Röntgen kan i ett första stadie ge diagnos av sjukdomen. Oftast behövs också en biopsi tas för fastställande av diagnos. Även provtagning av pleuravätskan kan ge värdefull diagnostisk information. Immunhistokemi är en viktig tilläggsanalys för diagnos med biopsier och cellblock. Immunhistokemi innebär färgning med antikroppar. För pleuralt mesoteliom kan exempelvis antikropparna PD-L1 och calretinin användas för diagnostisering. I denna studie färgades 10 parade pleurabiopsier och cellblock från pleuravätska från patienter med malignt mesoteliom. Antikropparna PD-L1 och calretinin användes. Syftet var att utvärdera uttrycket av PD-L1 och calretinin. Alla infärgade glas undersöktes i ljusmikroskop. För PD-L1 ansågs enbart membranfärgning som positivt infärgad och för calretinin ansågs cytoplasmatisk och/eller kärnfärgning som positiv. Procentuellt antal positiva tumörceller undersöktes vid två cutoff värden, ≥1 % och ≥10 %. Procentuellt antal <1 % vid cutoff ≥1 % och procentuellt antal <10 % vid cutoff ≥10 % ansågs negativt. Detta gällde för båda antikropparna. Antal positiva och negativa biopsier samt cellblock och konkordansen redovisades. Overall percentage agreement (OPA), Cohen’s kappa (κ) och konfidensintervall (CI) beräknades också. Resultatet visade på lägre antal positiva färgningar för cellblocken jämfört med biopsierna för båda antikropparna. För konkordansen visades att den vara lika för PD-L1 och calretinin vid cutoff ≥1 % men högre för PD-L1 än för calretinin vid cutoff ≥10 %. / Malignant mesothelioma is a rare and aggressive cancer. It is most common in the pleura, called pleural mesothelioma. The most common reason for developing pleural mesothelioma is exposure to asbestosis. Patients that get sick have a poor prognosis and limited treatment options. X-ray can in a first stadium give diagnose of the disease. A biopsy is often necessary to confirm the diagnosis. Even sampling of pleural fluid can give valuable diagnostic information. Immunohistochemistry is a common additional analysis for diagnosis with biopsy and cell block. Immunohistochemistry implies staining with antibodies. For example, for pleural mesothelioma the antibodies PD-L1 and calretinin can be used for diagnosis. In this study 10 paired pleural biopsies and cell blocks from pleural fluid from patients with malign mesothelioma were immunostained. The study aimed to evaluate expression of PD-L1 and calretinin. All the stained glasses were evaluated by a light microscope. For PD-L1, only membranous staining was considered positive and for calretinin cytoplasmic and/or nuclear staining was considered positive. Percentage number of positive tumor cells were evaluated at two cutoff values, ≥1 % and ≥10 %. Percentage number <1 % at cutoff ≥1 % and percentage number <10 % at cutoff ≥10 % were considered negative. The same criteria were applied for both antibodies. The number of positive and negative biopsies as well as cell blocks and the concordance were reported. Overall percentage agreement (OPA), Cohen’s kappa (κ) and confidence interval (CI) was also calculated. Result showed lower number of positive staining for the cell blocks than for the biopsies for both antibodies. The concordance was shown to be equal for PD-L1 and calretinin at cutoff ≥1 % but higher for PD-L1 than for calretinin at cutoff ≥10 %.

År 2020 stod lungcancer för högst andel cancer-relaterade dödsfall. Cellblockstekniken används inom diagnostik av vätskebaserad cytologi för att morfologiskt undersöka cellernas utseende och tumörcellers ursprung. I dagsläget upplever personal på Avdelningen för Klinisk patologi och cytologi Jönköping problematik vid klotformering av cellerna, vilket kan resultera i cellförlust. Syftet var att testa en ny metod för preparering av cellblock för lungcytologi samt utvärdera immunohistokemisk färgning med metoden. Studien behandlade 20 prover av pleuravätska och 20 lungcytologiprover som bearbetades till cellblock. Proteinglycerol och formaldehyd ersatte nuvarande agglutineringslösning bestående av koncentrerad formaldehyd och ättiksyra. Cellmängd beräknades på 39 snitt av två cytodiagnostiker, och graderades i tre nivåer från sparsam till riklig (1–3). Åtta cellblock selekterades för immunohistokemisk färgning. Det totala medelvärdet för cellularitet på snitten var måttlig (2,04). För pleuraproverna var medelvärdet 1,775 och för lungcytologiprover 2,315. Inga celler hittades på n=4 prover. Samtliga (n=8) immunohistokemiska färgningar med tillhörande kontroller godkändes av metodspecialist. Metoden ger snitt med tillräcklig cellularitet som fungerar väl med immunohistokemisk färgning. Metoden kan optimeras i relation till proteinförhållande och provtagning för att vid implementering bidra till en effektiviserad och individualiserad diagnostik. / In 2020, lung cancer accounted for the highest proportion of cancer-related mortalities. The cell-block technique is used in liquid-based cytology to morphologically examine the appearance of cells and the origin of tumour-cells.  Staff at the Department of Clinical pathology and cytology in Jönköping are experiencing problems with clot-formation of cells, which can result in cell-loss. The aim was to test a new method for preparing cell-blocks for lung cytology, and evaluate immunohistochemical staining with the method. The study treated 20 pleural fluids and 20 fine needle aspirations that were processed into cell-blocks. Protein-glycerol and formaldehyde replaced the current agglutination solution consisting of concentrated formaldehyde and acetic acid. The cell-count on 39 sections were graded by two Cytotechnologists in three levels from sparse to abundant (1-3). Eight cell-blocks were selected for immunohistochemical staining. The mean cellularity was moderate (2,04). For the pleural and lung cytology samples the mean values were 1,775 and 2.315. No cells were found on n=4 sections. Every (n=8) immunohistochemical staining with associated controls was approved by method specialist. The method provides sufficient cellularity and works with immunohistochemical staining. It can be optimized in relation to protein-ratio and sampling to contribute to streamlined and individualized diagnosis if implemented.