In coding theory we wish to find as many codewords as possible, while simultaneously maintaining high distance between codewords to ease the detection and correction of errors. For linear codes, this translates to finding high-dimensional subspaces of a given metric space, where the induced distance between vectors stays above a specified minimum. In this work I describe the recent advances of this problem in the contexts of lexicodes and Ferrers diagram rank-metric codes. In the first chapter, we study lexicodes. For a ring R, we describe a lexicographic ordering of the left R-module Rn. With this ordering we set up a greedy algorithm which sequentially selects vectors for which all linear combinations satisfy a given property. The resulting output is called a lexicode. This process was discussed earlier in the literature for fields and chain rings. We describe a generalization of the algorithm to finite principal ideal rings. In the second chapter, we investigate Ferrers diagram rank-metric codes, which play a role in the construction of subspace codes. A well-known upper bound for dimension of these codes is conjectured to be sharp. We describe several solved cases of the conjecture, and further contribute new ones. In addition, probabilities for maximal Ferrers diagram codes and MRD codes are investigated in a new light. It is shown that for growing field size, the limiting probability depends highly on the Ferrers diagram.

The Wilms’ tumor gene 1 (WT1) was first reported as a tumor suppressor gene in Wilms’ tumor. However, later studies have shown the oncogenic properties of WT1 in a variety of tumors. It was recently proposed that WT1 was a chameleon gene, due to its dual functions in tumorigenesis. We aimed to investigate the clinical significance of WT1 as biomarker in acute myeloid leukemia (AML) and clear cell renal cell carcinoma (ccRCC) and to elucidate the function of WT1 as an oncogene in squamous cell carcinoma of head and neck (SCCHN). In AML, it was suggested that WT1 expression was an applicable marker of minimal residual disease (MRD). In adult patients with AML, we found a good correlation between WT1 expression levels normalized to two control genes, β-actin and ABL. Outcome could be predicted by a reduction in WT1 expression in bone marrow (≥ 1-log) detected less than 1 month after diagnosis, when β-actin was used as control. Also, irrespective of the control gene used, outcome could be predicted by a reduction in WT1 expression in peripheral blood (≥ 2-log) detected between 1 and 6 months after treatment initiation. Previous studies in RCC demonstrated that WT1 acted as a tumor suppressor. Thus, we tested whether single nucleotide polymorphisms (SNPs) or mutations in WT1 might be associated with WT1 expression and clinical outcome in patients with ccRCC. We performed sequencing analysis on 10 exons of the WT1 gene in a total of 182 patient samples, and we identified six different SNPs in the WT1 gene. We found that at least one or two copies of the minor allele were present in 61% of ccRCC tumor samples. However, no correlation was observed between WT1 SNP genotypes and RNA expression levels. Moreover, none of the previously reported WT1 mutations were found in ccRCC. Nevertheless, we found that a favorable outcome was associated the homozygous minor allele for WT1 SNP. We then further investigated whether WT1 methylation was related to WT1 expression and its clinical significance. Methylation array and pyrosequencing analyses showed that the WT1 promoter region CpG site, cg22975913, was the most frequently hypermethylated CpG site. We found a trend that showed nearly significant correlation between WT1 mRNA levels and hypermethylation in the 5’-untranslated region. Hypermethylation in the WT1 CpG site, cg22975913, was found to be associated with patient age and a worse prognosis. One previous study reported that WT1 was overexpressed in SCCHN. That finding suggested that WT1 might play a role in oncogenesis. We found that both WT1 and p63 could promote cell proliferation. A positive correlation between WT1 and p63 expression was observed, and we identified p63 as a WT1 target gene. Furthermore, several known WT1 and p63 target genes were affected by knocking down WT1. Also, co-immunoprecipitation analyses demonstrated a protein interaction between WT1 and p53. In summary, WT1 gene expression can provide useful information for MRD detection during treatment of patients with AML. In RCC, our results suggested that the prognostic impact of WT1 SNPs was limited to the subgroup of patients that were homozygous for the minor allele, and that WT1 promoter hypermethylation could be used as a prognostic biomarker. In SCCHN, WT1 and p63 acted as oncogenes by affecting multiple genes involved in cancer cell growth.

Die akute lymphoblastische Leukämie (ALL) ist die häufigste maligne Erkrankung im Kindesalter. Trotz risikoadaptierter Chemotherapie erleiden 25 - 30 % der Kinder mit ALL ein Rezidiv. Im Rezidivfall liegen die Heilungschancen trotz intensivierter Therapie nur bei 35- 40%. In retrospektiven und prospektiven Analysen konnten wir einerseits prognostisch-relevante, genetische Merkmale in den Leukämiezellen von Kindern mit ALL-Rezidiv erstmals identifizieren. Diese leukämiespezifischen Merkmale umfassen sowohl Translokations-assoziierte Fusionsgene (TEL-AML1, BCR-ABL, MLL-Aberrationen, E2A-PBX1), Deletionen von Tumorsuppressorgenen (p15, p16, p18) als auch Mutationen in DNA-Reparaturgenen (NBS1). Zusammen mit der sensitiven molekularen Bestimmung der Kinetik der Leukämiezellreduktion (Reaktion auf die Therapie, (MRD, minimal residual disease)) läßt sich die Prognose der Kinder mit ALL-Rezidiv durch die Verwendung dieser Marker zuverlässiger bewerten. Andererseits zeigen unsere Analysen, dass das Ansprechen auf die Therapie und die Prognose von wirtseigenen Faktoren (medikamenten metabolisierenden Enzyme) und von der Interaktion zwischen Leukämie- und Stromazellen und löslichen Wachstumsfaktoren (Zytokinen) abhängt. Eine adäquatere Behandlung dieser Patientengruppe, die bereits eine intensive Therapie erhalten hat und wesentlich risikoreichere, mit einer hohen Akut- und Spättoxizität behaftete Therapieverfahren einschließt, lässt sich durch die Berücksichtigung dieser Ergebnisse erreichen. Die genetische Typisierung und die sensitive Quantifizierung des molekularen Ansprechens auf die Therapie ergänzen heute die klinischen Determinanten zur Risikostratifizierung der Kinder mit ALL-Rezidiv der derzeitigen Therapieoptimierungsstudie ALL-REZ BFM 2002 (Berlin-Frankfurt-Münster). / Acute lymphoblastic leukemia is the most common malignancy in childhood. Although the prognosis for pediatric ALL with risk-adapted chemotherapy has improved dramatically, 25-30% of the children suffer a relapse. The prognosis for relapsed ALL remains poor (35-40%). In retrospective and prospective studies, we identified prognostic-relevant genetic features in leukemic blasts from children with ALL relapse. These leukemia-specific aberrations include translocation-associated fusion genes (TEL-AML1, BCR-ABL, MLL changes, E2A-PBX1), deletions of tumor suppressor genes (p15, p16, p18) and point mutations in DNA repair genes (NBS1). Together with the sensitive quantitative assessment of the molecular response to therapy (MRD, minimal residual disease), prediction of outcome is now more reliable. Furthermore, response to therapy and, thus, prognosis is significantly dependent on modifying host factors (drug-metabolizing enzymes) and interactions between leukemic and stromal cells as well as soluble growth factors (cytokines). Today, the genetic characterization of leukemic cells as well as the molecular quantification of response to therapy complement prognostic significant clinical determinants allowing a more precise risk stratification of children with ALL relapse in the relapse trial ALL-REZ BFM 2002 of the BFM (Berlin-Frankfurt-Münster) study group.

Moisture induced damages to building envelopes can result in microbial growth possibly affecting the health and wellbeing of occupants. Recent failing structures and damaged buildings indicate a lack of tools to estimate risk of mould growth and moisture damage. In this work a so-called mould resistance design (MRD) model has been applied for mapping the risk for mould growth on a number of wood-containing wall structures. The MRD model introduces an engineering approach to moisture safety design in a similar way as for structural design, where load and resistance is compared. The MRD model introduces and quantifies the concepts of climatic exposure and material resistance and compares them through an MRD index. This MRD index incorporates a limit state, which gives the critical dose of exposure for a given resistance to initiate onset of mould growth.   Three conceptual wall structures were evaluated and analyzed in terms of MRD index: two wall systems with an air gap and one wall system without. A parametric study investigating the effect of parameter variation on MRD index was conducted. Evaluated parameters were: climate (geographic location), orientation, air changes per hour in the air gap, driving rain penetrating the facade layer, exterior plaster properties and wood type. The simulations were performed using the hygrothermal calculation software WUFI. The results indicate that the wall systems with a ventilated air gap performs better in terms of MRD index i.e. suggests a lower risk of initiation of mould growth than the wall system without air gap. The results of orientation variation show that wall systems perform differently dependent on layering structure. The inherent water sorption properties of the exterior plaster are shown to have a large effect on the results. In addition, uncertainties were found on how to accurately include hydrophobicity as a parameter in the model. The report concludes that geographical location and its specific climate is the most important parameter to consider when designing for moisture safety. The MRD model is recommended to be used in combination with traditional moisture safety evaluation.

Die akute myeloische Leukämie (AML) und das myelodysplastische Syndrom (MDS) stellen heterogene myeloische Neoplasien dar, deren Übergang in einander fließend ist. Während bei der AML ein Differenzierungsblock sowie eine unkontrollierte Proliferation myeloischer Vorläuferzellen dominieren, zeichnet sich das MDS vorrangig durch Dysplasien und variable Zytopenien, sowie ein erhöhtes Risiko der Transformation in eine AML aus. Trotz unseres zunehmenden Verständnisses dieser Erkrankungen ist die Prognose für AML und MDS Patienten noch immer häufig ungünstig. Auch mit der Entwicklung neuer zielgerichteter Therapiekonzepte behält die allogene hämatopoetische Stammzelltransplantation ihre große Bedeutung in der Therapie von AML und MDS. Insbesondere bei Patienten mit prognostisch ungünstigen Merkmalen stellt sie häufig die einzige kurative Therapieoption dar. Entsprechend kommt einer Risikostratifizierung bei Diagnose sowie im Krankheitsverlauf zur individuellen Therapieentscheidung eine große Bedeutung zu um personalisierte Behandlungen zu ermöglichen. Diese kann durch klinische Variablen, Immunphänotypisierung und zyto- oder molekulargenetische Veränderungen erfolgen. Diese Arbeit beschäftigt sich mit neuen klinischen, molekularen und durchflusszytometrischen Markern um die bestehenden Risikoklassifikationssysteme für Patienten mit AML und MDS weiter zu verfeinern und legt ein besonderes Augenmerk auf Patienten, die eine allogene Stammzelltransplantation erhalten. Der erste Abschnitt dieser Arbeit zeigt die prognostische Relevanz der leukämischen Stammzellpopulationen – definiert über die CD34+/CD38- Zellpopulation bzw. die GPR56 Expression - bei Diagnosestellung. Sowohl in der AML als auch im MDS scheint ein hoher Anteil leukämischer Stammzellen eine Subgruppe von Patienten mit ungünstiger Prognose unabhängig von aktuellen Risikostratifikationen und auch trotz Durchführung einer allogenen Stammzelltransplantation identifizieren zu können. Im zweiten Abschnitt wird ein Überblick über die zunehmende Relevanz der Risikostratifikation im Krankheitsverlauf - im Sinne der Bestimmung einer messbaren Resterkrankung (MRD) – erörtert und mit der Expressionshöhe von BAALC und MN1 mögliche neue Marker zur MRD Bestimmung vorgestellt. Der dritte Abschnitt zeigt, dass auch die Wahl des Konsolidierungskonzeptes die initiale molekulargenetische Risikostratifizierung beeinflussen kann. Während eine hohe Expression der AML-assoziierten Gene BAALC und MN1 bei AML Diagnose mit einer ungünstigen Prognose einhergeht, wenn die Patienten mit einer Chemotherapie behandelt werden, scheint eine allogene Stammzelltransplantation diesen prognostischen Einfluss aufzuheben. Außerdem wird die Relevanz einer allogenen Stammzelltransplantation bei älteren Patienten mit der prinzipiell eher günstigen molekulargenetischen Konstellation NPM1 mutiert/FLT3-ITD Wildtyp diskutiert. Im vierten Abschnitt wird der Einfluss klinischer Parameter auf die Prognose von AML und MDS Patienten diskutiert. Patienten mit einer sekundären (nach anderen myeloischen Erkrankungen) oder therapieassoziierten AML (nach vorausgegangener zytostatischer Therapie) weisen unter konventioneller Chemotherapie eine sehr ungünstige Prognose auf. Im Gegensatz dazu legen die Ergebnisse nahe, dass das Überleben nach allogener Stammzelltransplantation – wenn das genetische Risiko beachtet wird - nicht wesentlich schlechter ist als das von Patienten mit de novo AML. Außerdem wird gezeigt, dass sowohl das Vorliegen einer Adipositas zum Zeitpunkt der Diagnose einer AML als auch ein Gewichtsverlust zwischen Diagnose und allogener Stammzelltransplantation mit einer ungünstigen Prognose einhergehen. Zuletzt werden zwei Konditionierungsintensitäten vor Stammzelltransplantation in MDS Patienten verglichen und gezeigt, dass vor allem bei jüngeren Patienten intensivere Protokolle bessere Ergebnisse erzielen. Zusammenfassend zeigt diese Arbeit neue Möglichkeiten auf, die Risikostratifizierung für Patienten mit myeloischen Erkankungen sowohl bei Diagnose als auch im Krankheitsverlauf zu verbessern und leistet somit einen wichtigen Beitrag zur weiteren Personalisierung der Therapie von AML und MDS.:1. INTRODUCTION / EINLEITUNG 5 1.1 AML 5 1.1.1 DIAGNOSIS AND DISEASE CLASSIFICATION 5 1.1.2 RISK STRATIFICATION AT DIAGNOSIS 6 1.1.3 TREATMENT 8 1.2 MDS 9 1.2.1 DIAGNOSIS AND DISEASE CLASSIFICATION 9 1.2.2 RISK STRATIFICATION AT DIAGNOSIS 10 1.2.3 TREATMENT 12 1.3 ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION 13 2. FREQUENCY OF LEUKEMIA INITIATING CELL POPULATIONS AT DIAGNOSIS OF AML AND MDS 14 2.1 BACKGROUND 14 2.2 OWN CONTRIBUTION 14 Prognostic Impact of the CD34+/CD38- Cell Burden in Patients with Acute Myeloid Leukemia receiving Allogeneic Stem Cell Transplantation. Am J Hematol 2017; 92: 388-396. 15 High expression of the stem cell marker GPR56 at diagnosis identifies acute myeloid leukemia patients at higher relapse risk after allogeneic stem cell transplantation in context with the CD34+/CD38- population. Haematologica 2020. 105: 229260 15 The pre-treatment CD34+/CD38- cell burden as prognostic factor in MDS patients receiving allogeneic stem cell transplantation. Biol Blood Marrow Transplant 2019; 25: 1560-1566. 15 3 MEASURABLE RESIDUAL DISEASE DETECTION IN AML 37 3.1 BACKGROUND 37 3.2 OWN CONTRIBUTION 37 High Blood BAALC Copy Numbers at Allogeneic Transplantation Predict Early Relapse in Patients with Acute Myeloid Leukemia. Oncotarget 2017; 8:87944-87954. 38 Prognostic impact of blood MN1 copy numbers before allogeneic stem cell transplantation in patients with acute myeloid leukemia. HemaSphere 2019; 3: e167. 38 4. THE USE OF ALLOGENEIC HSCT IMPACTS THE RELEVANCE OF GENETIC PROGNOSTICATORS AT AML DIAGNOSIS 59 4.1 BACKGROUND 59 4.2 OWN CONTRIBUTION 59 Outcomes of older patients with NPM1 mutated and FLT3-ITD negative acute myeloid leukemia receiving allogeneic transplantation. HemaSphere 2020. 3; 4: e326. 59 Allogeneic stem cell transplantation mitigates the adverse prognostic impact of high diagnostic BAALC and MN1 expression in AML. Ann Hematol 2020. 99: 2417-2427. 60 5. CLINICAL AND TREATMENT-ASSOCIATED PROGNOSTICATORS IN AML AND MDS 76 5.1 BACKGROUND 76 5.2 OWN CONTRIBUTION 77 ELN risk stratification and outcomes in secondary and therapy-related AML patients consolidated with allogeneic stem cell transplantation. Bone Marrow Transplant. 2020 Nov 19. doi: 10.1038/s41409-020-01129-1. [Online ahead of print] 77 Nutritional status at diagnosis and weight changes impact outcomes in acute myeloid leukemia patients receiving allogeneic hematopoietic stem cell transplantation HemaSphere 2021. 5:2(e532). 77 Comparison of non-myeloablative and reduced-intensity allogeneic stem cell transplantation in older patients with myelodysplastic syndromes. Am J Hematol 2019; 94:1344-1352. 77 6. CONCLUSION / ZUSAMMENFASSUNG 106 7. REFERENCES / REFERENZEN 108 8. INDEX OF ABBREVIATIONS / INHALTSVERZEICHNIS 114 9. EIDESSTATTLICHE ERKLÄRUNG ZUR VORGELEGTEN HABILITATIONSSCHRIFT 116 10. COMPLETE LIST OF PUBLICATIONS / PUBLIKATIONSVERZEICHNIS 117 11. CURRICULUM VITAE / LEBENSLAUF 129 12. ACKNOWLEDGEMENTS / DANKSAGUNG 132

Background: For most patients with acute myeloid leukemia (AML) harboring a trisomy 8 an allogeneic hematopoietic stem cell transplantation (HSCT) is a suitable and recommended consolidation therapy. However, comparative outcome analyses between patients with and without trisomy 8 undergoing allogeneic HSCT have not been performed so far. Methods: We retrospectively analyzed clinical features, outcomes, and measurable residual disease (MRD) of 659 AML (12%, n = 81, with a trisomy 8) patients subjected to allogeneic HSCT as a consolidation therapy. Results: The presence of a trisomy 8 associated with a trend for higher age at diagnosis, AML of secondary origin, lower white blood cell counts at diagnosis, worse ELN2017 genetic risk, wild-type NPM1, and mutated IDH1/2 and JAK2. Outcomes after allogeneic HSCT in the entire cohort did not differ between patients with a sole trisomy 8, trisomy 8 with additional cytogenetic aberrations or without a trisomy 8. A trisomy 8 did not affect outcomes within the three ELN2017 risk groups. In accordance with findings in unselected patient cohorts, persistent MRD at allogeneic HSCT in patients with a trisomy 8 identified individuals with a higher risk of relapse following allogeneic HSCT. Conclusions: Outcomes of trisomy 8 patients after allogeneic HSCT did not compare unfavorably to that of other AML patients following allogeneic HSCT. Rather than the presence or absence of a trisomy 8, additional genetic aberrations and MRD at HSCT define outcome differences and aid in informed treatment decisions.

Submitted by admin tede (tede@pucgoias.edu.br) on 2017-02-06T16:52:19Z No. of bitstreams: 1 Caio Bruno Quinta de Souza Leal.pdf: 116273125 bytes, checksum: 5e247a988672d255dad21fa0aedcc5da (MD5) / Made available in DSpace on 2017-02-06T16:52:19Z (GMT). No. of bitstreams: 1 Caio Bruno Quinta de Souza Leal.pdf: 116273125 bytes, checksum: 5e247a988672d255dad21fa0aedcc5da (MD5) Previous issue date: 2014-01-31 / Monitoring of minimal residual disease (MRD) in patients with Chronic Myeloid Leukemia (CML) who receive treatment with Imatib mesylate (IM) is extremely important, because it enables the evaluation of the response to the treatment and the early diagnosis of possible recurrences. The objective of this study was to standardize molecular methods used in order to monitor MRD in patients with CML, on therapy with IM. Peripheral blood samples were collected from 11 patients diagnosed with CML in October 2012 to September 2013, in the Department of Hematology of Hospital Araújo Jorge of the Association to Combat Cancer in Goiás. Three months after starting treatment, patients underwent a new peripheral blood collection for evaluation of MRD. Detecting bcr-abl transcripts and endogenous controls (abl and β2m) employed reverse transcription methods associated with polymerase chain reaction (RT-PCR), while quantification of bcr-abl transcripts was achieved by using reverse transcription associated with real-time PCR (RQ-PCR) and Taqman probes. Specific oligonucleotides and probes recognizing e13a2 and e14a2 junctions of bcr-abl transcripts and to the abl endogenous control were used in this study. By the time of diagnosis, three patients (27.3%) expressed the b2a2 transcript, five patients (45.5%) expressed the b3a2 transcript, two patients (18.2%) expressed both transcripts and one patient (9%) did not express any of the transcripts. The endogenous controls analysis resulted in better amplification for the abl transcript, which was used in the RQ-PCR reactions. The assessment of DRM was possible in only eight patients, due to the loss of follow-up. Three months after starting treatment with IM, all patients presented complete hematologic response. However, only one patient (12.5%) presented the undetectable transcript, reaching the full molecular response, while the other seven patients (87.95%) presented MRD. One (12.5%) of the seven patients who presented MRD, reached complete molecular response, while six patients (75%) presented a reduction of two logs, achieving minor molecular response, and one patient (12.5%) presented only partial molecular response. By using molecular biology methods, our results have enabled the standardization and the establishment of a laboratory routine, according to the international guidelines, for monitoring MRD in patients with CML. / O monitoramento de doença residual mínima (DRM) em pacientes com leucemia meilóide crônica (LMC) que recebem tratamento com mesilato de imatibe (MI) é extremamente relevante, pois possibilita o acompanhamento da resposta e o diagnóstico precoce de eventuais recidivas da doença. O objetivo deste estudo foi padronizar métodos moleculares utilizados na avaliação de DRM em pacientes com LMC, em tratamento com MI. Amostras de sangue periférico foram coletadas de 11 pacientes diagnosticados com LMC, no período de outubro de 2012 a setembro de 2013, no Setor de Hematologia do Hospital Araújo Jorge da Associação de Combate ao Câncer em Goiás. Três meses após o início do tratamento, os pacientes foram submetidos a uma nova coleta de sangue periférico para avaliação de DRM. A detecção dos transcritos bcr-abl e controles endógenos (abl e β2m) empregaram os métodos de transcrição reversa associados à reação em cadeia da polimerase (RT-PCR), enquanto a quantificação dos transcritos bcr-abl foi feita por meio de transcrição reversa associada à PCR em tempo real (RQ-PCR), utilizando a metodologia de sondas de hidrólise (TaqMan). Oligonucleotídeos e sondas Taqman específicos para as junções e13a2 e e14a2 dos transcritos bcr-abl e para o controle endógeno (abl) foram usados neste estudo. Ao diagnóstico, três pacientes (27,3%) expressaram o transcrito b2a2, cinco pacientes (45,5%) o transcrito b3a2, dois pacientes (18,2%) expressaram ambos os transcritos e um paciente (9%) não expressou nenhum dos transcritos. A amplificação dos controles endógenos resultou em melhor amplificação para o transcrito abl, que foi usado nas reações de RQ-PCR. A avaliação de DRM foi possível em oito pacientes, devido à perda de seguimento dos demais. Três meses após o início do tratamento com MI, todos os pacientes apresentaram resposta hematológica completa. No entanto, apenas um paciente (12,5%) apresentou o transcrito bcr-abl indetectável, alcançando a reposta molecular completa, enquanto os outros sete pacientes (87,95%) apresentaram DRM. Dentre os sete pacientes que apresentaram DRM, seis (75%) apresentaram redução de um a dois logs, alcançando resposta molecular menor, enquanto um (12,5%) apresentou resposta molecular parcial. Nossos resultados possibilitaram a padronização e o estabelecimento de uma rotina segundo as diretrizes internacionais, para monitoramento da DRM em pacientes com LMC, utilizando métodos de biologia molecular.

Made available in DSpace on 2016-08-10T10:38:55Z (GMT). No. of bitstreams: 1 CAIO BRUNO QUINTA DE SOUZA LEAL - PARTE 1.pdf: 34304765 bytes, checksum: 476dda15021939ae8be633403ae9dabe (MD5) Previous issue date: 2014-01-31 / Monitoring of minimal residual disease (MRD) in patients with Chronic Myeloid Leukemia (CML) who receive treatment with Imatib mesylate (IM) is extremely important, because it enables the evaluation of the response to the treatment and the early diagnosis of possible recurrences. The objective of this study was to standardize molecular methods used in order to monitor MRD in patients with CML, on therapy with IM. Peripheral blood samples were collected from 11 patients diagnosed with CML in October 2012 to September 2013, in the Department of Hematology of Hospital Araújo Jorge of the Association to Combat Cancer in Goiás. Three months after starting treatment, patients underwent a new peripheral blood collection for evaluation of MRD. Detecting bcr-abl transcripts and endogenous controls (abl and β2m) employed reverse transcription methods associated with polymerase chain reaction (RT-PCR), while quantification of bcr-abl transcripts was achieved by using reverse transcription associated with real-time PCR (RQ-PCR) and Taqman probes. Specific oligonucleotides and probes recognizing e13a2 and e14a2 junctions of bcr-abl transcripts and to the abl endogenous control were used in this study. By the time of diagnosis, three patients (27.3%) expressed the b2a2 transcript, five patients (45.5%) expressed the b3a2 transcript, two patients (18.2%) expressed both transcripts and one patient (9%) did not express any of the transcripts. The endogenous controls analysis resulted in better amplification for the abl transcript, which was used in the RQ-PCR reactions. The assessment of DRM was possible in only eight patients, due to the loss of follow-up. Three months after starting treatment with IM, all patients presented complete hematologic response. However, only one patient (12.5%) presented the undetectable transcript, reaching the full molecular response, while the other seven patients (87.95%) presented MRD. One (12.5%) of the seven patients who presented MRD, reached complete molecular response, while six patients (75%) presented a reduction of two logs, achieving minor molecular response, and one patient (12.5%) presented only partial molecular response. By using molecular biology methods, our results have enabled the standardization and the establishment of a laboratory routine, according to the international guidelines, for monitoring MRD in patients with CML. / O monitoramento de doença residual mínima (DRM) em pacientes com leucemia meilóide crônica (LMC) que recebem tratamento com mesilato de imatibe (MI) é extremamente relevante, pois possibilita o acompanhamento da resposta e o diagnóstico precoce de eventuais recidivas da doença. O objetivo deste estudo foi padronizar métodos moleculares utilizados na avaliação de DRM em pacientes com LMC, em tratamento com MI. Amostras de sangue periférico foram coletadas de 11 pacientes diagnosticados com LMC, no período de outubro de 2012 a setembro de 2013, no Setor de Hematologia do Hospital Araújo Jorge da Associação de Combate ao Câncer em Goiás. Três meses após o início do tratamento, os pacientes foram submetidos a uma nova coleta de sangue periférico para avaliação de DRM. A detecção dos transcritos bcr-abl e controles endógenos (abl e β2m) empregaram os métodos de transcrição reversa associados à reação em cadeia da polimerase (RT-PCR), enquanto a quantificação dos transcritos bcr-abl foi feita por meio de transcrição reversa associada à PCR em tempo real (RQ-PCR), utilizando a metodologia de sondas de hidrólise (TaqMan). Oligonucleotídeos e sondas Taqman específicos para as junções e13a2 e e14a2 dos transcritos bcr-abl e para o controle endógeno (abl) foram usados neste estudo. Ao diagnóstico, três pacientes (27,3%) expressaram o transcrito b2a2, cinco pacientes (45,5%) o transcrito b3a2, dois pacientes (18,2%) expressaram ambos os transcritos e um paciente (9%) não expressou nenhum dos transcritos. A amplificação dos controles endógenos resultou em melhor amplificação para o transcrito abl, que foi usado nas reações de RQ-PCR. A avaliação de DRM foi possível em oito pacientes, devido à perda de seguimento dos demais. Três meses após o início do tratamento com MI, todos os pacientes apresentaram resposta hematológica completa. No entanto, apenas um paciente (12,5%) apresentou o transcrito bcr-abl indetectável, alcançando a reposta molecular completa, enquanto os outros sete pacientes (87,95%) apresentaram DRM. Dentre os sete pacientes que apresentaram DRM, seis (75%) apresentaram redução de um a dois logs, alcançando resposta molecular menor, enquanto um (12,5%) apresentou resposta molecular parcial. Nossos resultados possibilitaram a padronização e o estabelecimento de uma rotina segundo as diretrizes internacionais, para monitoramento da DRM em pacientes com LMC, utilizando métodos de biologia molecular

Analysis of donor chimerism has become a routine method for the documentation of engraftment after allogeneic hematopoietic stem cell transplantation (HSCT). In recent years several groups have also focused on the application of this technique for the detection of relapsing disease after allogeneic HSCT. This review addresses technical issues (sensitivity, specificity) and discusses the advantages and limitations of methods currently used for chimerism analysis and their usefulness for the detection of MRD. In addition, the potential impact of novel procedures, e.g. subset chimerism or real-time PCR-based procedures, is discussed. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

The phenomenon of fading constitutes a fundamental problem in wireless communications. Researchers have proposed many methods to improve the reliability of communication over wireless channels in the presence of fading. Many studies on this topic have focused on diversity techniques. Transmit diversity is a common diversity type in which multiple antennas are employed at the transmitter. Space-time coding is a technique based on transmit diversity introduced by Tarokh et alii in 1998. In this thesis, various types of space-time codes are examined. Since they were originally introduced in the form of trellis codes, a major part is devoted to space-time trellis codes where the fundamental design criteria are established. Then, space-time block coding, which presents a different approach, is introduced and orthogonal spacetime block codes are analyzed in some detail. Lastly, rank codes from coding theory are studied and their relation to space-time coding are investigated.