Instruction of haematopoietic lineage choices, evolution of transcriptional landscapes and cancer stem cell hierarchies derived from an AML1-ETO mouse model.

The t(8;21) chromosomal translocation activates aberrant expression of the AML1-ETO (AE) fusion protein and is commonly associated with core binding factor acute myeloid leukaemia (CBF AML). Combining a conditional mouse model that closely resembles the slow evolution and the mosaic AE expression pattern of human t(8;21) CBF AML with global transcriptome sequencing, we find that disease progression was characterized by two principal pathogenic mechanisms. Initially, AE expression modified the lineage potential of haematopoietic stem cells (HSCs), resulting in the selective expansion of the myeloid compartment at the expense of normal erythro- and lymphopoiesis. This lineage skewing was followed by a second substantial rewiring of transcriptional networks occurring in the trajectory to manifest leukaemia. We also find that both HSC and lineage-restricted granulocyte macrophage progenitors (GMPs) acquired leukaemic stem cell (LSC) potential being capable of initiating and maintaining the disease. Finally, our data demonstrate that long-term expression of AE induces an indolent myeloproliferative disease (MPD)-like myeloid leukaemia phenotype with complete penetrance and that acute inactivation of AE function is a potential novel therapeutic option.

Comparação entre métodos de estimativa da evapotranspiração de referência (ETo) em regiões citrícolas paulistas

As regiões escolhidas para o estudo são importantes no cenário citrícola irrigado, tanto em expansão quanto em atuação. Os dados meteorológicos utilizados para as estimativas são integrantes do banco de dados climáticos do software IRRIPLUS®. Com este software, estimou-se a evapotranspiração de referência (ETo) diária, utilizando as metodologias propostas por Penman-Monteith (EToPM), Hargreaves & Samani (EToHG) e Blaney-Criddle-FAO (EToBC), para o período de um ano. O objetivo deste estudo foi comparar os métodos de estimativa com EToPM, utilizando parâmetros de regressão (β0, β1), coeficiente de determinação (r²), e correlação (r), estimativa do erro-padrão (SEE), índices de concordância (d) e confiança (c). Os resultados indicam tendência de EToHG à superestimativa dos valores de ETo, exceto na região de Jaú, e subestimativa por EToBC, para todas as regiões estudadas. Nos valores mensais de ETo, os três métodos apresentam a mesma distribuição. O método de EToHG apresenta-se como uma excelente alternativa na estimativa de consumo de água pela cultura.

Estimation methods of reference evapotranspiration (ETo) for Uberlândia -MG

One approach to verify the adequacy of estimation methods of reference evapotranspiration is the comparison with the Penman-Monteith method, recommended by the United Nations of Food and Agriculture Organization - FAO, as the standard method for estimating ET0. This study aimed to compare methods for estimating ET0, Makkink (MK), Hargreaves (HG) and Solar Radiation (RS), with Penman-Monteith (PM). For this purpose, we used daily data of global solar radiation, air temperature, relative humidity and wind speed for the year 2010, obtained through the automatic meteorological station, with latitude 18° 91' 66" S, longitude 48° 25' 05" W and altitude of 869m, at the National Institute of Meteorology situated in the Campus of Federal University of Uberlandia - MG, Brazil. Analysis of results for the period were carried out in daily basis, using regression analysis and considering the linear model y = ax, where the dependent variable was the method of Penman-Monteith and the independent, the estimation of ET0 by evaluated methods. Methodology was used to check the influence of standard deviation of daily ET0 in comparison of methods. The evaluation indicated that methods of Solar Radiation and Penman-Monteith cannot be compared, yet the method of Hargreaves indicates the most efficient adjustment to estimate ETo.

Estimativa da evapotranspiração de referência (ETo padrão FAO), para Minas Gerais, na ausência de alguns dados climáticos

A equação para determinar a evapotranspiração de referência (ETo), método de Penman-Monteith, parametrizada pela FAO (PM-FAO56), requer dados que, muitas vezes, não estão disponíveis na maioria das estações climatológicas. Para superar o problema da disponibilidade de dados climáticos, o boletim 56 da FAO propõe vários procedimentos para estimar a ETo na ausência dos dados de radiação, umidade relativa e velocidade do vento, e posterior substituição. Baseado nisto, o objetivo do presente estudo foi comparar a ETo estimada na ausência de dados de velocidade do vento ou da umidade relativa ou radiação solar com a ETo estimada, com os dados completos para várias localidades de Minas Gerais. A comparação foi realizada utilizando o coeficiente angular da regressão, o coeficiente de determinação, o índice de concordância de Willmott e os erros associados a cada metodologia. Verificou-se que, na ausência de dados de umidade relativa e velocidade do vento, o método de Penman-Monteith apresenta pequenos erros de estimativa. Na ausência dos dados de radiação, os erros são maiores em relação às outras variáveis. Na disponibilidade apenas das temperaturas máximas e mínimas, o método de PM-FAO56 apresenta desempenho superior ao método de Hargreaves-Samani.

ETO-Tuotekehitysprojektin suorituskyvyn mittaaminen

Tutkimuksen tavoitteena oli selvittää, mitä suorituskyvyn osa-alueita ETO (Engineer To Order) – tyyppinen tuotekehitysprojekti pitää sisällään, ja mitkä niistä ovat projektin onnistumisen kannalta tärkeimpiä, eli niin sanottuja menestystekijöitä. Edelleen näiden tunnistettujen menestystekijöiden pohjalta oli tavoitteena kehittää suorituskyvyn mittausmalli, jonka avulla voisi arvioida ja ohjata projektin ja projektinhallinnan suorituskykyä. Tärkeimmät suorituskyvyn näkökulmat tässä mallissa olivat projektin kannattavuuden -, asiakastyytyväisyyden - ja projektinhallinnan näkökulmat. Malli tehtiin rautatieliikenne toimialalla toimivalle yritykselle, joka asetti omat vaatimuksensa mallille. Tutkimus tehtiin kolmivaiheisesti. Ensimmäisessä vaiheessa tutustuttiin aiempiin tutkimuksiin, joiden pohjalta pyrittiin löytämään projektin menestystekijät, sekä näiden suorituskyvyn analysointiin toimiviksi todetut mittausmenetelmät. Ensimmäisessä vaiheessa tutustuttiin myös alaa ohjaaviin standardeihin, ja niiden asettamiin erityisvaatimuksiin projektien suorituskyvyn mittaukselle. Tutkimuksen toinen vaihe oli tapaustutkimus kohdeyrityksen sisäiselle projektiorganisaatiolle, joka toteutettiin survey-kyselynä. Kyselyn avulla pyrittiin löytämään ne menestystekijät, jotka projektiorganisaation itsensä mielestä olivat kaikkein tärkeimpiä projektin onnistumisen kannalta. Kyselyllä pyrittiin myös selvittämään sidosryhmien välisiä näkemyseroja projektin menestystekijöistä. Kysymykset muodostettiin synteesinä tutkimuksen ensimmäisessä vaiheessa löydetyistä menestystekijöistä ja standardien vaatimista pakollisista mitattavista tekijöistä. Tärkeimmäksi menestystekijäksi kyselyssä nousi projektipäällikön kompetenssi. Kyselyssä vastaajilta kysyttiin myös sitä, minkä menestystekijöiden suorituskykyä heidän mielestään tulisi mitata. Tärkeimmäksi mitattavaksi osa-alueeksi nousi aikataulun tarkka seuranta. Kolmannessa vaiheessa näiden kohdeyrityksen menestystekijöiden pohjalta luotiin mittausmalli, joka pyrkii mittaamaan projektin ja projektinhallinnan suorituskykyä tasapainoisesti valituista näkökulmista. Malli ottaa huomioon erityisesti projektinhallinnan suorituskyvyn vaikutuksen projektin lopputuotteena syntyvien hyödykkeiden laatuun. Kohdeyrityksen kokonaissuorituskyvyn kannalta on erittäin tärkeää parantaa projektien suorituskykyä, johon projektien suorituskyvyn mittaaminen on ratkaisu. Standardien asettamat vaatimukset antavat hyvät lähtökohdat kokonaisvaltaisen mittausjärjestelmän luomiselle.

Comparação entre métodos de estimativa da evapotranspiração de referência (ETo) em regiões citrícolas paulistas

As regiões escolhidas para o estudo são importantes no cenário citrícola irrigado, tanto em expansão quanto em atuação. Os dados meteorológicos utilizados para as estimativas são integrantes do banco de dados climáticos do software IRRIPLUS®. Com este software, estimou-se a evapotranspiração de referência (ETo) diária, utilizando as metodologias propostas por Penman-Monteith (EToPM), Hargreaves & Samani (EToHG) e Blaney-Criddle-FAO (EToBC), para o período de um ano. O objetivo deste estudo foi comparar os métodos de estimativa com EToPM, utilizando parâmetros de regressão (β0, β1), coeficiente de determinação (r²), e correlação (r), estimativa do erro-padrão (SEE), índices de concordância (d) e confiança (c). Os resultados indicam tendência de EToHG à superestimativa dos valores de ETo, exceto na região de Jaú, e subestimativa por EToBC, para todas as regiões estudadas. Nos valores mensais de ETo, os três métodos apresentam a mesma distribuição. O método de EToHG apresenta-se como uma excelente alternativa na estimativa de consumo de água pela cultura.

Interpolação geoespacial da evapotranspiração de referência (ETo) em regiões com escassez de dados: estudo de caso no Sul de Minas Gerais, Brasil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estimation methods of reference evapotranspiration (ETo) for Uberlândia -MG

One approach to verify the adequacy of estimation methods of reference evapotranspiration is the comparison with the Penman-Monteith method, recommended by the United Nations of Food and Agriculture Organization - FAO, as the standard method for estimating ET0. This study aimed to compare methods for estimating ET0, Makkink (MK), Hargreaves (HG) and Solar Radiation (RS), with Penman-Monteith (PM). For this purpose, we used daily data of global solar radiation, air temperature, relative humidity and wind speed for the year 2010, obtained through the automatic meteorological station, with latitude 18° 91' 66 S, longitude 48° 25' 05 W and altitude of 869m, at the National Institute of Meteorology situated in the Campus of Federal University of Uberlandia - MG, Brazil. Analysis of results for the period were carried out in daily basis, using regression analysis and considering the linear model y = ax, where the dependent variable was the method of Penman-Monteith and the independent, the estimation of ET0 by evaluated methods. Methodology was used to check the influence of standard deviation of daily ET0 in comparison of methods. The evaluation indicated that methods of Solar Radiation and Penman-Monteith cannot be compared, yet the method of Hargreaves indicates the most efficient adjustment to estimate ETo.

In vivo consequences of AML1-ETO fusion protein expression for hematopoiesis

The t(8;21) (q22;q22) translocation fusing the ETO (also known as MTG8) gene on human chromosome 8 with the AML1 (also called Runx1 or CBFα) gene on chromosome 21 is one of the most common genetic aberrations found in acute myeloid leukemia (AML). This chromosomal translocation occurs in 12 % of de novo AML cases and in up to 40 % of the AML-M2 subtype of the French-American-British classification. To date, the in vivo function of aberrant AML1-ETO fusion protein expression has been investigated by several groups. However, in these studies, controversial results were reported and some key issues remain unknown. Importantly, the consequences of aberrant AML1-ETO expression for self-renewing hematopoietic stem cells (HSCs), multipotent hematopoietic progenitors (MPPs) and lineage-restricted precursors are not known. rn The aim of this thesis was to develop a novel experimental AML1-ETO in vivo model that (i) overcomes the current lack of insight into the pre-leukemic condition of t(8;21)-associated AML, (ii) clarifies the in vivo consequences of AML1-ETO for HSCs, MPPs, progenitors and more mature blood cells and (iii) generates an improved mouse model suitable for mirroring the human condition. For this purpose, a conditional tet on/off mouse model expressing the AML1-ETO fusion protein from the ROSA26 (R26) locus was generated. rn Aberrant AML1-ETO activation in compound ROSA26/tetOAML1-ETO (R26/AE) mice caused high rates of mortality, an overall disruption of hematopoietic organs and a profound alteration of hematopoiesis. However, since the generalized activity of the R26 locus did not recapitulate the leukemic condition found in human patients, it was important to restrict AML1-ETO expression to blood cell lineages. Therefore, bone marrow cells from non-induced R26/AE mice were adoptively transplanted into sublethal irradiated RAG2-/- recipient mice. First signs of phenotypical differences between AML1-ETO-expressing and control mice were observed after eight to nine months of transgene induction. AML1-ETO-expressing mice showed profound changes in hematopoietic organs accompanied by manifest extramedullary hematopoiesis. In addition, a block in early erythropoiesis, B- and T-cell maturation was observed and granulopoiesis was significantly enhanced. Most interestingly, conditional activation of AML1-ETO in chimeric mice did not increase HSCs, MPPs, common lymphoid precursors (CLPs), common myeloid progenitors (CMPs) and megakaryocyte-erythrocyte progenitors (MEPs) but promoted the selective amplification of granulocyte-macrophage progenitors (GMPs). rn The results of this thesis provide clear experimental evidence how aberrant AML1-ETO modulates the developmental properties of normal hematopoiesis and establishes for the first time that AML1-ETO does not increase HSCs, MPPs and common lineage-restricted progenitor pools but specifically amplifies GMPs. The here presented mouse model not only clarifies the role of aberrant AML1-ETO for shaping hematopoietic development but in addition has strong implications for future therapeutic strategies and will be an excellent pre-clinical tool for developing and testing new approaches to treat and eventually cure AML.rn

Rolle der gemeinsamen Expression von AML1-ETO und weiteren Onkogenen für die Induktion von Leukämie

Akute Leukämien treten in allen Altersstufen auf. Akute lymphatische Leukämie (ALL) ist die häufigste Leukämie bei Kindern, während akute myeloischen Leukämien (AML) mit verschiedenen Untergruppen etwa 80% aller akuten Leukämien bei Erwachsenen ausmachen. Die Translokation t(8;21) resultiert in der Entstehung des Fusionsgens AML1-ETO und zählt zu den häufigen Translokationen bei der AML. Dabei fusioniert die DNA-bindende Domäne des AML1 mit dem fast kompletten ETO-Protein. AML1-ETO wirkt als dominanter Repressor der AML1-vermittelten transkriptionellen Regula-tion wichtiger hämatopoetischer Zielgene. Klinische Daten legen nahe, dass trotz der klarer Assoziation zwischen AML und der t(8;21) Translokation bei AML Patienten zusätzliche genetische Veränderungen – so genannte ‚second hits‘ – notwendig sind, um eine Leukämie effizient zu induzieren. Klinisch relevanten Komplimentationsonkogene sind unter anderen die aktivierte Rezeptortyrosinkinase FLT3, JAK2, NRAS, KRAS, c- KIT.rnZiel der vorliegenden Arbeit war es, ein Mausmodell zu etablieren, welches humane akute myeloische Leukämie rekapituliert und bei dem die Expression der entsprechen-den Onkogene reguliert werden kann. Als erstes wurde untersucht, ob eine gemeinsame Expression von AML1-ETO mit kRASG12D zur Induktion von Leukämie führen kann. Hierfür wurden Tiere generiert die gemeinsam AML1-ETO und kRASG12D unter der regulatorischen Sequenz des Tetrazyklin-Operators exprimierten. Der große Vorteil dieser Technologie ist die regulierbare Reversibilität der Genexpression. Um die Ex-pression der Zielgene auf blutbildende Zellen zu beschränken, wurden Knochenmark-chimären hergestellt. Im Beobachtungszeitraum von 12 Monaten führte die Expression von AML1-ETO und AML1-ETO/kRASG12D nicht zur Induktion einer akuten Leukä-mie. Die normale hämatopoetische Entwicklung war jedoch in diesen Tieren gestört. Der beobachtete Phänotyp entsprach einem myelodysplastischen Syndrome (MDS).rnIm zweiten Ansatz, wurden Tiere generiert die gemeinsam AML1-ETO und FLT3-ITD exprimierten. Hierfür wurden hämatopoetische Stammzellen aus ROSA26-iM2/tetO-AML1-ETO isoliert und mit Hilfe des retroviralen Vektors mit FLT3-ITD transduziert. In diesem Modell war es möglich, in kurzer Zeit eine akute Leukämie mit zu induzieren. Einige wenige Tiere hatten zum Zeitpunkt des Todes Anzeichen einer biphänotypischen Leukämie mit lymphatischen und myeloischen Blastenpopulationen. In drei Tieren in-duzierte die alleinige Expression von FLT3-ITD eine Leukämie. Alle Leukämien wurden durch FACS, Zytologie und Histopathologie bestätigt. Knochenmark- bzw. Milzzellen aus den erkrankten Tieren waren in der Lage nach Transfer in sekundäre Rezipienten eine Leukämie auszulösen. Somit besaßen sie ein uneingeschränktes Selbsterneue-rungspotential.rnEin erster Versuch, in dem AML1-ETO Expression in leukämischen Zellen abgeschaltet und FLT3-ITD mit Tyrosinkinase-Inhibitor inhibiert wurde, zeigte keine wesentliche Veränderung in der Leukämieprogression.rnDieses Leukämiemodell erlaubt die Rolle der beteiligten Onkogene während verschie-dener Stadien der Leukämie zu erforschen und damit möglicherweise neue Ansätze für Therapiestrategien zu entwickeln.