Regressões lineares em geocronologia: isócronas, errócronas e pseudoisócronas

Several methods of linear regression reported in the literature for use in geochonology are discussed in order to guide geochronologists in the selection of an adequate model and also to distinguish among isochrons, errorchrons and pseudoisochrons. For Rb/Sr isochron methods, at least 6 models or statistical treatments are available, each one with its peculiarities. Based in two examples, we show that for cases when the experimental data points scatter within limits determined by the experimental errors, and using or not the weighted regression treatment, the resultant parameters are essentially concordant. The estimates of associated error, however especially for initial ratios, show variations by up to a factor of 7, depending upon the statistical procedure. In view of this fact and on the basis of simulated test data, we propose not only more adequate models, but also general criteria that must be observed in the selection of samples in order to obtain an isochron. If the data to be regressed show a scatter in excess of experimental error for geological reasons (open system), four regression models are available to treat the parameters defined in these cases as errorchrons. These models are briefly discussed in terms of their applications. In the Pb/U methods, in which the usual regression treatment is applied to obtain the best-fit line, some additional peculiarities are pointed outDiversos métodos de regressão linear propostos na literatura, para fins geocronológicos, são discutidos de modo a orientar os geocronólogos na escolha de modelos adequados e também distinguir entre isócronas, errócronas ou pseudoisócronas. No caso do método isocrôni co Rb/Sr, são disponíveis pelo menos 6 modelos ou abordagens estatísticas, cada qual com peculiaridades específicas. Exemplifica-se o caso em que os pontos se alinham dentro dos erros experimentais e utilizando modelos, adequados ou não, que empregam a técnica de pondera ção dos pontos, os parâmetros resultantes são praticamente concordantes, porém os erros podem variar até por um fator tão alto quanto 7, conforme a formulação estatística proposta. Em vista deste fato, propõem-se, com base também em exemplos, os modelos mais adequados e também critérios gerais que devem ser observados até na seleção de amostras para a obtenção de isócronas. Nos casos em que os pontos não se alinham dentro dos erros experimentais, por causas geológicas (sistema aberto), são abordados 4 dos modelos sugeridos e é feita também uma discussão em que casos seriam aplicáveis. No método da concórdia Pb/U, em que as regressões usuais também são aplicáveis para determinar a melhor reta, algumas particularidades adicionais são discutida

BALB/c Mice Infected with Antimony Treatment Refractory Isolate of Leishmania braziliensis Present Severe Lesions due to IL-4 Production

Leishmaniasis is a neglected disease that affects more than 12 million people worldwide. In Brazil, the cutaneous disease is more prevalent with about 28,000 new cases reported each year, and L. braziliensis is the main causative agent. The interesting data about the infection with this parasite is the wide variety of clinical manifestations that ranges from single ulcerated lesions to mucocutaneous and disseminated disease. However, experimental models to study the infection with this parasite are difficult to develop due to high resistance of most mouse strains to the infection, and the mechanisms underlying the distinct manifestations remain poorly understood. Here, the authors use a mouse experimental model of infection with different L. braziliensis isolates, known to induce diseases with distinct severity in the human hosts, to elucidate immune mechanisms that may be involved in the different manifestations. They showed that distinct parasite isolates may modulate host response, and increased IL-4 production and Arg I expression was related to more severe disease, resulting in longer length of disease with larger lesions and reduced parasite clearance. These findings may be useful in the identification of immunological targets to control L. braziliensis infection and potential clinical markers of disease progression

Characterization of the macrophage transcriptome in glomerulonephritis-susceptible and -resistant rat strains

Crescentic glomerulonephritis (CRGN) is a major cause of rapidly progressive renal failure for which the underlying genetic basis is unknown. WKY rats show marked susceptibility to CRGN, while Lewis rats are resistant. Glomerular injury and crescent formation are macrophage-dependent and mainly explained by seven quantitative trait loci (Crgn1-7). Here, we used microarray analysis in basal and lipopolysaccharide (LPS)-stimulated macrophages to identify genes that reside on pathways predisposing WKY rats to CRGN. We detected 97 novel positional candidates for the uncharacterised Crgn3-7. We identified 10 additional secondary effector genes with profound differences in expression between the two strains (>5-fold change, <1% False Discovery Rate) for basal and LPS-stimulated macrophages. Moreover, we identified 8 genes with differentially expressed alternatively spliced isoforms, by using an in depth analysis at probe-level that allowed us to discard false positives due to polymorphisms between the two rat strains. Pathway analysis identified several common linked pathways, enriched for differentially expressed genes, which affect macrophage activation. In summary, our results identify distinct macrophage transcriptome profiles between two rat strains that differ in susceptibility to glomerulonephritis, provide novel positional candidates for Crgn3-7, and define groups of genes that play a significant role in differential regulation of macrophage activity

MicroRNA miR-125b causes leukemia

MicroRNA miR-125b has been implicated in several kinds of leukemia. The chromosomal translocation t(2;11)(p21;q23) found in patients with myelodysplasia and acute myeloid leukemia leads to an overexpression of miR-125b of up to 90-fold normal. Moreover, miR-125b is also up-regulated in patients with B-cell acute lymphoblastic leukemia carrying the t(11;14)(q24;q32) translocation. To decipher the presumed oncogenic mechanism of miR-125b, we used transplantation experiments in mice. All mice transplanted with fetal liver cells ectopically expressing miR-125b showed an increase in white blood cell count, in particular in neutrophils and monocytes, associated with a macrocytic anemia. Among these mice, half died of B-cell acute lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, or a myeloproliferative neoplasm, suggesting an important role for miR-125b in early hematopoiesis. Furthermore, coexpression of miR-125b and the BCR-ABL fusion gene in transplanted cells accelerated the development of leukemia in mice, compared with control mice expressing only BCR-ABL, suggesting that miR-125b confers a proliferative advantage to the leukemic cells. Thus, we show that overexpression of miR-125b is sufficient both to shorten the latency of BCR-ABL–induced leukemia and to independently induce leukemia in a mouse model.National Institutes of Health (U.S.) (Grant DK068348)National Institutes of Health (U.S.) (grant 5P01 HL066105

Usefulness of long-distance inverse polymerase chain reaction for molecular detection of 14q32 translocation in a clinical setting.

All mature B-cell leukaemias and lymphomas have a clonal Ig gene recombination, and half of them have a reciprocal chromosomal translocation involving the 14q32 locus. The 14q32 translocation partners are variable, such as BCL-2, BCL-1 and BCL-6, thus accounting for the difficulty in molecular detection by the current genomic polymerase chain reaction (PCR) method. To identify B-cell clones efficiently with an Ig gene rearrangement and reciprocal inter-chromosomal translocation, we verified the usefulness, in a practical laboratory setting, of our modified long-distance inverse (LDI) PCR method for detecting IgH gene rearrangements involving inter- and intra-chromosomal segments. The total run time of this LDI PCR method was 5.5 h. Using 24 samples of mature B-cell leukaemias and lymphomas, the modified LDI PCR gave clonally rearranged amplicons in 83 % (20/24) of cases. Direct sequencing results of the amplicons revealed inter-chromosomal translocations in 5 cases (25 %) and intra-chromosomal rearrangements in the remaining 15 cases (75 %). The partners of the inter-chromosomal translocation consisted of the 11q13.3 segment containing a partial BCL1 sequence in 3 cases; 18q21.3 segment containing a partial BCL2 sequence in one case; and a segment of 7q11.2 in one case. We present an LDI PCR-based methodology for the efficient identification of 14q32 translocations, with modifications to reduce the total run time to within one day

Detection of translocations affecting the BCL6 locus in B cell non-Hodgkin's lymphoma by interphase fluorescence in situ hybridization

Structural alterations in 3q27 affecting the BCL6 locus are among the most frequent changes in B-NHL. The aim of the present study was to establish an interphase-FISH assay for the detection of all diverse BCL6 translocations in B-NHL. Two different approaches were tested, one using a PAC-clone spanning the major breakpoint region (MBR) of BCL6 (span-assay), and another using two BAC clones flanking the MBR (flank-assay). Interphase FISH with the span-assay detected the various BCL6 translocations in seven B-NHL cell lines. The dual-color flank-assay was evaluated in two laboratories independently: in normal controls, the cutoff level for false-positive signals was 2.6%, whereas the cutoff level for false-negatives in the seven cell lines was 7.5%. To test the feasibility of the FISH strategies, 30 samples from patients with B-NHL with cytogenetic abnormalities of 3q27 were evaluated with both assays. In 21 cases, the span-assay indicated a BCL6 rearrangement. In 18 of the 21 cases, the dual-color flank-assay confirmed the translocation including 12 different partner chromosomal loci. The three false-positive cases detected with the span-assay showed trisomy of chromosome 3 by cytogenetic analyses, and they were correctly classified as non-rearranged with the flank-assay. In summary, our FISH strategy using two differently labeled flanking BCL6 BAC probes provides a robust, sensitive, and reproducible method for the detection of common and uncommon abnormalities of BCL6 gene in interphase nuclei. The routine application of this assay to patients with B-NHL will allow the assessment of the diagnostic and prognostic significance of BCL6 rearrangements