Genome-wide association analysis with selective genotyping identifies candidate loci for adult height at 8q21.13 and 15q22.33-q23 in Mongolians

We performed a genome-wide association study with 23,465 microsatellite markers to identify genes related to adult height. Selective genotyping was applied to extremely tall and extremely short individuals from the Khalkh-Mongolian population. Two loci, 8q21.13 and 15q22.33, which showed the strongest association with microsatellites were subjected to further analyses of SNPs in 782 tall and 773 short individuals. The most significant association was observed with SNP rs2220456 at 8q21.13 (P = 0.000016). In the LD block at 15q22.32, SNP rs8038652 located in intron 1 of IQCH was strongly associated (P = 0.0003), especially the AA genotype of the SNP under a recessive model was strongly associated with adult height (P = 0.000046)

Dimorfismo, uso de hábitat y dieta de Anolis Maculiventris (Lacertilia: Dactyloidae), en bosque pluvial tropical del chocó, Colombia

Se estudiaron algunos aspectos de la biología poblacional de Anolis maculiventris, en zonas de bosque pluvial tropical de la región natural chocoana. Los machos y hembras presentaron una coloración café claro con un pequeño lunar en la parte posterior de la cabeza, las proporciones de sexo en los individuos capturados mostraron diferencias estadísticas significativas (X2 : 4,81, gl: 1, p: 0,028). El grado de dimorfismo sexual (GDS) e índice de dimorfismo sexual fue de 1,08 mm y un de 8,5 mm respectivamente. Las hembras difieren significativamente de los machos en todas las regiones corporales medidas. Se disectaron 30 individuos a los cuales se les determino su estado reproductivo, en donde la talla mínima de madures en hembras (LHA) fue de 30,7 mm, con un máximo de 39,9 mm y una media de 36,5 mm. En machos la talla mínima de madurez fue de 30,4 mm, y máxima de 36,7 mm, con una media de 33,8 mm; lo cual indica que las hembras fueron más grandes que los machos. Se evaluó el uso de hábitat, mostrando diferencias estadísticas significativas entre los sustrato utilizados (X2 =16,89, gl: 3, p: 0,001); y para las posiciones verticales mostraron el mismo comportamiento (X2 =3,77, gl: 3, p: 0,207). Para el análisis dietario de la especie se disectaron 30 estómagos, de los cuales 15 pertenecieron a machos y 15 a hembras, se registróun total 46 presas gastrointestinales que componen su dieta mostrando a los órdenes Himenóptera (Formicidae) y Coleóptera como los de mayor importancia relativa en la dieta.We studied some aspects of the population biology of A. maculiventris, in areas of tropical rainforest in the natural region of Choco. The males and females brown coloration with a small mole on the back of the head, the proportions and gender in the captured individuals showed significant statistical differences (X2 : 4,81, gl: 1, p: 0,028). The degree of sexual dimorphism (GDS) and sexual dimorphism index was 1.08, mm and 8.5 mm respectively. Females differ significantly from males in all the parts of the body measures. 30 Individuals have spread, which was determined to their reproductive status and failed to register a size minimum of maturity (LHA) in females is 30.7 mm, with a maximum of 39.9 mm and an average of 36.5 mm. In the male the minimum maturity sexual 30.4 mm, and a maximum of 36.7 mm, with an average of 33.8 mm; which indicates that females were more large than males. Assessed habitat use, showing significant statistical differences between the used substrate (X2 =16,89, gl: 3, p: 0,001); and for vertical positions, showed the same behavior (X2 =3,77, gl: 3, p: 0,207). For analyzing dietary species spread 30 stomach, of which 15 belonged to males and 15 females, will register a total 46 gastrointestinal dams that compose its diet showing the orders Coleoptera and Hymenoptera (Formicidae) as the of greater relative importance in the diet

The induction of myeloid differentiation in the APL cells induced by ATRA treatment <i>in vitro</i> and <i>in vivo</i>.

<p>(<b>A</b>) Cytospin slides of the induced APL cells cultured with or without 1 µM ATRA for 6 days. The mature neutrophils with Auer rods are seen in the ATRA-treated group. The images were captured with a BX41 microscope (Olympus). The bar indicates 10 µm. (<b>B</b>) The differential cellular counts of the induced APL cells cultured with or without ATRA. The average leukocyte differentiation in three independent experiments is shown. (<b>C</b>) The results of an immunofluorescent analysis of the PML distribution in the induced APL cells before and after treatment with ATRA. The images were captured with an LSM510 META confocal microscope (Carl Zeiss). All the bars indicate 10 µm. (<b>D, E</b>) The induction of myeloid differentiation in the induced APL cells by ATRA <i>in vivo</i>. The secondary recipients transplanted with the induced APL cells were then intraperitoneally treated with ATRA for 21 days. Cytospin slides of EGFP⁺/hCD45⁺/hCD33⁺ cells from the secondary recipients transplanted with the induced APL cells are shown (<b>D</b>). The images were captured with a BX41 microscope (Olympus). The bar indicates 10 µm. Their differentiated cellular counts were evaluated, and the representative series data are indicated (<b>E</b>).</p

Establishment of humanized APL <i>in vivo</i>.

<p>(<b>A</b>) The proportion of EGFP⁺/human CD45⁺ cells in the bone marrow of leukemic NOG mice. Each dot represents a single mouse. The horizontal line represents the median value. (<b>B</b>) The proportion of CD33⁺ cells among the EGFP⁺/human CD45⁺ cells in the bone marrow of the leukemic NOG mice. Each dot represents a single mouse. The horizontal line represents the median value. (<b>C</b>) The expression of <i>PML-RARA</i> in RT-PCR was detected only in the EGFP⁺ fraction obtained from the engrafted human CD45⁺/CD33⁺ cells. The cells were obtained from bone marrow 16 weeks after transplantation. <i>B2M, beta 2 microglobulin</i>. The <i>PML-RARA</i> expression vector and human CD34⁺ cells were used as a positive control for the <i>PML-RARA</i> and <i>B2M</i> analysis, respectively. (<b>D</b>) The differential counts of the engrafted CD45⁺/CD33⁺/EGFP⁺ and EGFP⁻ cells from the mice transplanted with <i>PML-RARA</i>-induced human CD34⁺ cells. They were obtained from bone marrow 16 to 20 weeks after transplantation. The data represent the means. (<b>E</b>) A representative photograph of the resulting leukemic cells which morphologically recapitulated APL. Faggot cells were recognized, as seen in the right top corner. The images were captured with a BX41 microscope (Olympus). The bar indicates 10 µm. (<b>F</b>) The results of a Southern blot analysis of the genomic DNA from induced APL cells with an EGFP probe. Clonal bands are shown by arrow heads: white, seen in both BM and SP; orange, seen only in BM or SP. BM, bone marrow; SP, spleen. (<b>G</b>) The leukemia-free duration in NOG mice transplanted with <i>PML-RARA-</i>transduced CD34⁺ cells.</p

APL-LIC in the humanized APL <i>in vivo</i> model.

<p>(<b>A</b>) The engraftment of the induced APL cells in the secondary recipients. The bone marrow cells were obtained 16 weeks after transplantation and evaluated. Each dot represents a single mouse. (<b>B</b>) The immunophenotype of the induced APL cells in the secondary recipients. The representative pattern is shown. (<b>C</b>) The engraftment capacity in each fraction from the induced APL cells in the secondary recipients. The bone marrow cells were obtained 16 weeks after transplantation and evaluated. (<b>D</b>) The engraftment of the CD34⁻ fraction in the secondary recipients. The engrafted cells are shown as EGFP⁺/CD45⁺ cells.</p

The function of <i>PML-RARA</i> in human CD34⁺ cells <i>in vitro</i>.

<p>(<b>A</b>) The results of an immunofluorescent analysis of PML distribution in human CD34⁺ cells transduced with <i>PML-RARA</i>. The images were captured with an LSM510 META confocal microscope (Carl Zeiss). The bars indicate 10 µm. (<b>B</b>) The colony-forming assay using <i>PML-RARA</i>-transduced CD34⁺ cells. The cells were sorted by EGFP expression 48 h after infection. Colony formation was evaluated on days 10 to 12 after plating the cells. The average numbers of colonies from three independent experiments are shown. Data represent the means ± SD. The asterisks (*) indicate p<0.05. (<b>C</b>) The proportion of each kind of colony was calculated from the results of the colony-forming assay shown in (B). The percentages of CFU-GM are higher in <i>PML-RARA</i>-infected cells than in control (MIGR1) cells (p = 0.013).</p

Characteristics of the induced APL <i>in vivo</i>.

<p>(<b>A</b>) The representative expression pattern of cell surface markers in the induced APL cells determined by a flow cytometric analysis. All the scatter plots show the relationships between the EGFP positivity and cell surface marker expression. The whole living cells gated as a propidium iodide-negative fraction in the bone marrow were analyzed. The induced APL cells were recognized as EGFP⁺ cells. A few murine hematopoietic cells, recognized as a human CD45⁻ fraction, were detected in this mouse. (<b>B</b>) The heat map of the microarray analysis using the 510 APL-specific genes for the comparison of the induced APL cells (purple, n = 3) with APL (red, n = 16), other types of AML (M0, 1, 2 and 4 in FAB classification, green, n = 62) and normal promyelocytes (blue, n = 5) in a previous study <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111082#pone.0111082-Payton1" target="_blank">[26]</a>. The red triangles (n = 2) for a total 16 APL cases show the clinical APL samples whose microarray data were obtained in this study.</p

<i>PML-RARA</i> targeted human common myeloid progenitors for APL leukemogenesis.

<p>(<b>A</b>) The sorting strategy for CD34⁺/CD38⁻, CD34⁺/CD38⁺ and CD34⁻/CD33⁺ cells. Human cord blood was first separated into CD34⁺ and CD34⁻ cells by magnetic beads, and then sorted into three fractions by a FACS vantage instrument. (<b>B</b>) The expression of <i>PML-RARA</i> mRNA in each of the fractions after retroviral transfection. <i>B2M, beta 2 microglobulin</i>. The <i>PML-RARA</i> expression vector was used as a positive control for the <i>PML-RARA</i> analysis. (<b>C</b>) A colony-forming assay using <i>PML-RARA</i>-transduced CD34⁺/CD38⁺ and CD34⁺/CD38⁻ cells. The average of three independent experiments is shown. The data represent the means ± SD. (<b>D</b>) The total numbers of colonies of <i>PML-RARA</i>-transduced CD34⁺/CD38⁺ and CD34⁺/CD38⁻ cells shown in (C) are highlighted. The data represent the means ± SD (n = 3). (<b>E</b>) The development of the induced APL from CD34⁺/CD38⁺ cells in NOG mice. Each sorted fraction from human cord blood, as seen in (A), was retrovirally transduced with <i>PML-RARA</i> and transplanted into irradiated NOG mice. The percentages were determined by the frequency of EGFP⁺/CD45⁺/CD33⁺ cells at 16 to 20 weeks after transplantation. Each dot represents a single mouse. The horizontal line represents the median value. (<b>F</b>) The sorting strategy for common myeloid progenitors (CMP), granulocyte-monocytic progenitors (GMP), and megakaryocyte-erythrocyte progenitors (MEP). Human cord blood was separated into CD34⁺ cells by magnetic beads, CD34⁺/CD38⁺ cells were sorted out, and were finally divided into CMP, GMP and MEP by the FACS vantage instrument. (<b>G</b>) The transduction efficiency of <i>PML-RARA</i> in CMP, GMP and MEP. Representative data are shown. (<b>H</b>) The development of the induced APL from the human hematopoietic progenitors in NOG mice. Each sorted progenitor fraction from human cord blood, as seen in (F), was retrovirally transduced with <i>PML-RARA</i> and transplanted into irradiated NOG mice. The percentages were determined by the frequency of EGFP⁺/CD45⁺/CD33⁺ cells at 16 to 20 weeks after transplantation. Each dot represents a single mouse. The horizontal line represents the median value.</p