Analysis of the Leukemogenic Potential of the CALM/AF10 Fusion Gene in Patients, Transgenic Mice and Cell Culture Models

The t(10;11)(p13;q14) is a recurring translocation resulting in the fusion of the CALM and AF10 genes. The leukemogenic CALM/AF10 fusion genes codes for a 1595 amino acids protein. This translocation was first identified in a patient with hystiocytic lymphoma and was subsequently found in patients with AML, T-ALL and malignant lymphoma. This translocation is found in younger patients and is associated with a poor prognosis. The CALM/AF10-associated leukemias can exhibit myeloid, lymphoid or mixed lymphoid-meyloid features, indicating a stem cell or an early commited progenitor as the target cell of leukemic transformation. At the present time the target cells in CALM/AF10-associated leukemogenesis are unknown. It is also not known which target genes are up or downregulated by the presence of the CALM/AF10 fusion protein. To answer these questions, the following experiments were performed: 1) Five transgenic mouse lines, two expressing CALM/AF10 under the control of the immunoglobulin heavy chain enhancer promoter and three under the control of the murine proximal Lck promoter were generated. Although the CALM/AF10 expression was confirmed to be present and specific to the cells targeted by the promoters used (B- and T-cell progenitors for IgH and Lck promoters, respectively), the transgenic animals did not show a phenotype that could be detected after meticulous clinical, haematological, immunological, flow cytometrical and immunohistopatological analysis . 2) We performed molecular characterization of several CALM/AF10 patient samples: A group of 13 patients with different types of leukemia: case 1 (AML M2), case 2 (Acute Biphetnotypic leukemia), case 3 (Pre T-ALL), case 4 (Acute Undifferentiated Leukemia), case 5 (PreT-ALL), cases 6 and 7 (ProT-ALL), case 8 (T-ALL), case 9 (AML), case 14 (T-ALL), case 15, 16 and 17 (AML) with a t(10;11) translocation detected by cytogenetic analysis suggesting a CALM/AF10-rearrangement. The samples were analyzed for the presence of the CALM/AF10 and AF10/CALM fusion transcripts by RT-PCR and sequence analysis. All these patients were found to be positive for the CALM/AF10 fusion. In addition, we analyzed a series of twenty-nine patients with T-ALL with T-cell receptor ≥¥ rearrangement. Among these patients, four (case 10 to 13) were positive for the CALM/AF10 fusion transcript, indicating a high incidence of CALM/AF10 fusions in this group of leukemia. Three different breakpoints in CALM at nucleotide 1926, 2091 and a new exon, with 106 bases inserted after nt 2064 of CALM in patient 4 were found. In AF10 four breakpoints were identified: at nucleotide position 424, 589, 883 and 979. In patient 16 we found an extra exon before nt 424 of AF10. In seven patients it was also possible to amplify the reciprocal AF10/CALM fusion transcript (case 1, 3, 4, 8, 9, 10 and 14). There was no correlation between disease phenotype and breakpoint location. Ten CALM/AF10 positive patients were analyzed using oligonucleotide microarrays representing 33,000 different genes (U133 set, Affymetrix). Analysis of microarray gene expression signatures of these patients revealed high expression levels of the polycomb group gene BMI1, the homeobox gene MEIS1 and the HOXA cluster genes HOXA1, HOXA4, HOXA5, HOXA7, HOXA9, and HOXA10. The overexpression of HOX genes seen in these CALM/AF10 positive leukemias is reminiscent to the pattern seen in leukemias with rearrangements of the MLL gene, normal karyotypes and complex aberrant karyotypes suggesting a common effector pathway (i.e. HOX gene deregulation) for these diverse leukemias. In addition, the general pattern of gene expression of CALM/AF10 patients when compared to other leukemia subtypes and to normal bone marrow was dominated by a global downregulation of genes some of them with function identified as related to important molecular mechanisms, such as membrane trafficking, cell growth regulation, proliferation, differentiation and tumor suppression. 3) We cloned CALM/AF10 fusion gene into a vector that allowed us to induce the expression of CALM/AF10 using doxycycline in transiently and stably-transfected NIH3T3 and HEK293 cells. This system will be an important tool to identify direct CALM/AF10 target genes and to answer the question whether continued CALM/AF10 expression is necessary to maintain the CALM/AF10-associated expression pattern

Accurate cosmic shear errors: do we need ensembles of simulations?

Accurate inference of cosmology from weak lensing shear requires an accurate shear power spectrum covariance matrix. Here, we investigate this accuracy requirement and quantify the relative importance of the Gaussian (G), super-sample covariance (SSC) and connected non-Gaussian (cNG) contributions to the covariance. Specifically, we forecast cosmological parameter constraints for future wide-field surveys and study how different covariance matrix components affect parameter bounds. Our main result is that the cNG term represents only a small and potentially negligible contribution to statistical parameter errors: the errors obtained using the G+SSC subset are within lesssim 5% of those obtained with the full G+SSC+cNG matrix for a Euclid-like survey. This result also holds for the shear two-point correlation function, variations in survey specifications and for different analytical prescriptions of the cNG term. The cNG term is that which is often tackled using numerically expensive ensembles of survey realizations. Our results suggest however that the accuracy of analytical or approximate numerical methods to compute the cNG term is likely to be sufficient for cosmic shear inference from the next generation of surveys

Availability of floral resources in yellow passion fruit cultivars

The reproductive strategies and resources available in the flower are characteristics that stimulate research studies on the genetic improvement and fruit yield of commercial cultivars. In this sense, this study aimed to study the floral biology aspects of passion fruit cultivars in Tangará da Serra region, Mato Grosso State, Brazil at different evaluation times. The experiment was carried out with eight cultivars of yellow passion fruit from an ex situ germplasm bank of the State University of Mato Grosso. The floral resources (pollen and nectar) were evaluated in a completely randomized, factorial arrangement (8 cultivars x 5 evaluation times), with five replications. Throughout anthesis, the pollen viability, stigma receptivity, and sugar volume and concentration in the nectar were evaluated. The evaluations were carried out in 1-h intervals, between 1:30 and 5:30 p.m. The highest nectar volume production was at 2:30 p.m., decreasing gradually thereafter at the later evaluation times, for all cultivars. Pollen viability increased from the first to the second evaluation time and then decreased gradually until the last evaluation. Stigma receptivity was higher than 90% in all cultivars, at all evaluation times. BRS Rubi do Cerrado and FB 200 were the most promising cultivars for nectar volume and concentration, pollen viability, and stigma receptivity characteristics.The reproductive strategies and resources available in the flower are characteristics that stimulate research studies on the genetic improvement and fruit yield of commercial cultivars. In this sense, this study aimed to study the floral biology aspects of passion fruit cultivars in Tangará da Serra region, Mato Grosso State, Brazil at different evaluation times. The experiment was carried out with eight cultivars of yellow passion fruit from an ex situ germplasm bank of the State University of Mato Grosso. The floral resources (pollen and nectar) were evaluated in a completely randomized, factorial arrangement (8 cultivars x 5 evaluation times), with five replications. Throughout anthesis, the pollen viability, stigma receptivity, and sugar volume and concentration in the nectar were evaluated. The evaluations were carried out in 1-h intervals, between 1:30 and 5:30 p.m. The highest nectar volume production was at 2:30 p.m., decreasing gradually thereafter at the later evaluation times, for all cultivars. Pollen viability increased from the first to the second evaluation time and then decreased gradually until the last evaluation. Stigma receptivity was higher than 90% in all cultivars, at all evaluation times. BRS Rubi do Cerrado and FB 200 were the most promising cultivars for nectar volume and concentration, pollen viability, and stigma receptivity characteristics

Identification and characterization of OSTL (RNF217) encoding a RING-IBR-RING protein adjacent to a translocation breakpoint involving ETV6 in childhood ALL

Genomic aberrations involving ETV6 on band 12p13 are amongst the most common chromosomal abnormalities in human leukemia. The translocation t(6;12)(q23;13) in a childhood B-cell acute lymphoblastic leukemia (ALL) cell line fuses ETV6 with the putative long non-coding RNA gene STL. Linking STL properties to leukemia has so far been difficult. Here, we describe a novel gene, OSTL (annotated as RNF217 in Genbank), which shares the first exon and a CpG island with STL but is transcribed in the opposite direction. Human RNF217 codes for a highly conserved RING finger protein and is mainly expressed in testis and skeletal muscle with different splice variants. RNF217 shows regulated splicing in B cell development, and is expressed in a number of human B cell leukemia cell lines, primary human chronic myeloid leukemia, acute myeloid leukemia with normal karyotype and acute T-ALL samples. Using a yeast two-hybrid screen, we identified the anti-apoptotic protein HAX1 to interact with RNF217. This interaction could be mapped to the C-terminal RING finger motif of RNF217. We propose that some of the recurring aberrations involving 6q might deregulate the expression of RNF217 and result in imbalanced apoptosis signalling via HAX1, promoting leukemia development

Juntos: grupo de apoio em educação para a diversidade

Anais do 35º Seminário de Extensão Universitária da Região Sul - Direitos Humanos e JustiçaO Grupo Juntos objetiva estudar, acompanhar e dar suporte às demandas de ensino, pesquisa e extensão no que se refere à diversidade. A diversidade está relacionada a questões de etnicidadade, equidade de gênero, livre expressão de identidades sociais, diversidade sexual e religiosa. Juntos busca afirmar-se como uma política de acesso e permanência dentro de um projeto educacional inclusivo, entendendo que, para tanto, a boa convivência é fundamental. A ação se organiza em 3 frentes: a) Políticas de Acesso, b) Políticas de Permanência, c) Direitos Humanos e Diversidad

Dinâmicas de surgimento de novos partidos na América do Sul (1979-2015)

O início da década de 1980 caracteriza-se pelo esgotamento dos regimes autoritários na América do Sul. O longo processo de transição que passaram alguns países, ou a abrupta ruptura institucional de outros, marcou o retorno dos partidos à arena de disputa política. Como diversos autores pontuaram, desde então, tem-se assistido a emergência de novos atores políticos ou transformações de partidos anteriormente consolidados. Neste trabalho, investigamos as dinâmicas do surgimento de novos partidos no subcontinente. Os mesmos são tratados enquanto organizações que se apresentam pela primeira vez em pleito nacional. Consideramos todas as eleições (91) realizadas no período de 1979 até 2015, no sentido de observar as variações do número de novos partidos que surgiram em cada pleito, a partir de fontes secundárias (outros trabalhos) e primárias (bancos de dados dos respectivos tribunais eleitorais). A literatura que trata do surgimento de novos partidos, em democracias consolidadas ou emergentes, aponta distintas variáveis que impactam nos incentivos à gênese de novas organizações. Crises políticas ou sociais, atores políticos e/ou incentivos institucionais. Em nosso caso, a partir da análise estatística multivariada, buscamos perceber quais são as variáveis que têm mais efeito sobre o surgimento de novos partidos. Os resultados apontam que existem diferenças significativas entre os países, sendo os Andinos muito mais propensos à emergência de novos partidos, devido a alterações Constitucionais específicas. Causas sociais, como alto desemprego e baixo crescimento econômico, também foram significantes para explicar por que em alguns contextos surgiram mais novos partidos do que em outros

Morphological and molecular characterization of native Heliconia sp. accessions of the Amazon region

Heliconias are tropical plants with ornamental potential. These plants are particularly used in the floriculture industry because of their exotic colors and shapes. Species characterization is important for the selection of genotypes for the ornamental plant market and subsequent application in studies of genetic improvement. The aim of this study was to estimate the genetic divergence of Heliconia densiflora and Heliconia psittacorum accessions based on quantitative morphological and molecular markers. The mor- phological and molecular descriptors revealed genetic variability among the accessions evaluated. The greatest genetic variability was observed among H. psittacorum accessions, whose sample number was also larger compared to H. densiflora. Morphological characterization was efficient in differentiating the two Heliconia species, especially to characteristics such as bract and inflores- cence length, postharvest durability, and flower stem diameter, which contributed most to the divergence in this study. On the other hand, molecular characterization identified one H. densiflora individual that was grouped with the H. psittacorum genotypes. The results showed that ISSR markers can differentiate closely related H. densiflora and H. psittacorum individuals. The materials evaluated can contribute to the maintenance of local genetic diversity through the germplasm bank of the local breeding program of ornamental tropical plants

Interplay of weak ferromagnetism, ferroelasticity and shape-memory effects in the spin-orbit coupled antiferromagnet K2_2ReCl6_6

The magnetic and structural phase transitions occurring in K$_2$ReCl$_6$ were studied by macroscopic and microscopic techniques. Structural phase transitions associated with rotations of the ReCl$_6$ octahedra lower the symmetry from cubic to monoclinic, form ferroelastic domains, and are visible in susceptibility, specific heat and thermal expansion measurements. In the antiferromagnetically ordered state slightly below $T_{\rm N}$=12\,K these domains can be rearranged by a magnetic field inducing a relative elongation of the polydomain crystal parallel to the field of 0.6\%. At zero field the magnetic structure in K$_2$ReCl$_6$ does not exhibit a weak ferromagnetic component, but at large magnetic field a distinct magnetic structure with a finite weak ferromagnetic component is stabilized. High magnetic fields rearrange the domains in the crystal to align the weak ferromagnetic moment parallel to the field. The altered domain structure with the crystal elongation is abruptly suppressed at lower temperature but persists upon heating to well above $T_{\rm N}$. However, heating above the lowest structural phase transition and successive cooling restore the initial shape, i.e. a magnetic shape memory effect.Comment: 11 pages, 9 figure

Secondary pollen presentation and foral traits of Heliconia psittacorum

The secondary presentation of pollen consists of a foral mechanism where the presentation of pollen occurs in other foral structures in addition to the anther, in order to increase the precision of the dispersion of the pollen by the vectors. This study aims to describe the temporal dynamics of secondary pollen presentation, and morphological and morphometric characteristics in stages of pre- anthesis and anthesis in genotypes of fve natural Heliconia psittacorum populations. For the study of foral morphometry traits of length of the fower, stamen, stigma and height of flament insertion in the petal in bud and fower were measured. The foral morphology of pre-anthesis buds and fowers in anthesis, the presence or absence of characteristics such as herkogamy, region of flament insertion in the petal, region of stylar hairs, and of secondary pollen deposition were evaluated. Treatments of controlled pollinations, self-pollination, geitonogamy, cross-pollination, natural pollination and growth to pollen tube were sampled. Floral herkogamy and pollen transfer to the adhered hairs in the stylar region were clearly observed during anthesis, constituting the frst record of occurrence of secondary pollen presentation in Heliconiaceae. Pollen tube growth was inhibited in the stigmatic, style and basal regions of the pistil. Natural fruiting produced little or no fruit. The positioning of the stamens above the stigma, pollen viability and stigma receptivity during anthesis of H. psittacorum fowers may favor self-pollination. The stylar hairs observed in all H. psittacorum populations’ help in the retention of pollen grains. The low fruiting rate in controlled and natural pollinations suggest that the main propagation form of H. psittacorum in the study areas is based on asexual reproduction