Characteristics of Family Dynamics among Japanese Families in Japan

The purpose of this study is to identify family dynamics and their relationships to selected socio-demographic characteristics and mental status among Japanese families in Japan. The Family Dynamics Measure II (FDM II) and a socio-demographic questionnaire were used to obtain data. Participants (n=195; male n=62, female n=129, gender unknown n=4) were obtained from a city, western outskirts of Tokyo. Majority of respondents were married (n=165, 84.6%) and with children (n=128, 65.6%). Family size ranged from 2 to 9 persons (mean= 3.66, SD=1.42). Three-generation families, I.e. presences of grandparent(s), daughter(s)-in-law, and parent(s)-in-law (n=25, 12.8%) were small in numbers. When there were family members ages over 70, female respondents showed negative relations to 4 of 6 dimensions of FDM II. Further, when there were family members ages between 12-17, both male and female respondents showed negative relationships to 3 of 6 dimensions of FDM II. Mental status showed a positive relationship to age of male respondents. Nurses should be aware of relationships between affecting factors and dimensions of family dynamics. The understanding of these gender differences among males and females and the conditions of the family could be quite helpful when advising or counseling family members for betterment of family health

Pediatric AML: From Biology to Clinical Management

Pediatric acute myeloid leukemia (AML) represents 15%–20% of all pediatric acute leukemias. Survival rates have increased over the past few decades to ~70%, due to improved supportive care, optimized risk stratification and intensified chemotherapy. In most children, AML presents as a de novo entity, but in a minority, it is a secondary malignancy. The diagnostic classification of pediatric AML includes a combination of morphology, cytochemistry, immunophenotyping and molecular genetics. Outcome is mainly dependent on the initial response to treatment and molecular and cytogenetic aberrations. Treatment consists of a combination of intensive anthracycline- and cytarabine-containing chemotherapy and stem cell transplantation in selected genetic high-risk cases or slow responders. In general, ~30% of all pediatric AML patients will suffer from relapse, whereas 5%–10% of the patients will die due to disease complications or the side-effects of the treatment. Targeted therapy may enhance anti-leukemic efficacy and minimize treatment-related morbidity and mortality, but requires detailed knowledge of the genetic abnormalities and aberrant pathways involved in leukemogenesis. These efforts towards future personalized therapy in a rare disease, such as pediatric AML, require intensive international collaboration in order to enhance the survival rates of pediatric AML, while aiming to reduce long-term toxicity

A neonate with a unique non-Down syndrome transient proliferative megakaryoblastic disease

Transient myeloproliferative disorder (TMD) is a leukemia type that occurs typically in newborns. In Down syndrome, TMD is referred to as transient abnormal myelopoiesis (TAM).(32) Recently, transientness has also been reported in acute myeloid leukemia patients with germline trisomy 21 mosaicism, and even in cases with somatic trisomy 21, with or without GATA1 mutations. TMD cases without trisomy 21 are rare, and recurrent genetic aberrations that aid in clinical decision-making are scarcely described. We describe here a TMD patient without trisomy 21 or GATA1 mutation in whom single-nucleotide polymorphism analysis of leukemic blasts revealed a novel combined submicroscopic deletion (5q31.1-5q31.3 and 8q23.2q24)

2009

ABSTRACT © F e r r a t a S t o r t i F o u n d a t i o n by controlling transcriptional regulators, such as GATA1 and RUNX1. Methods Patients' samples Samples were provided by the Dutch Childhood Oncology Group (DCOG, the Netherlands), the AML-Berliner-FrankfurtMünster Study Group (Germany and Czech Republic), the SaintLouis Hospital (France), and the Royal Hospital for Sick Children (UK). Isolation of genomic DNA and total cellular RNA was performed using Trizol reagent. Each study group provided morphological and cytogenetic classification and clinical data. Institutional review board approval for these studies had been obtained in the participating centers. Detection of deletions and mutations of IKZF1 Multiplex ligation-dependent probe amplification (MLPA) was performed using the p335-B1;ALL-IKZF1 kit (MRC Holland, Amsterdam, the Netherlands; data available at http: //www.mlpa.com). The data were analyzed with GeneMarker v. 1.85 (SoftGenetics, State College, Pennsylvania, USA). Data were normalized to reference probes and control samples. A deletion was defined as a peak ratio below 0.75; an amplification was defined as a peak ratio above 1.25. Array comparative genomic hybridization (Array-CGH) was performed using the human genome CGH Microarray 105K (Agilent Technologies, Santa Clara, California, USA) according to the manufacturer's protocol; data were analyzed with Genomic Workbench v. 5.0.14 (Agilent Technologies, Santa Clara, California, USA). Direct sequencing was used to analyze mutations or frameshifts in exons 4, 5 and 8, which are found to be hotspot areas of mutations in pediatric BCP-ALL (primers are listed in Online 13,28 Cytogenetic and molecular characterization Reverse-transcription-PCR was performed on hotspot areas as previously described by Balgobind et al., Gene expression profiling Gene expression profiling (GEP) data were available from earlier studies. 30 Quantitative reverse-transcription-PCR (qRT-PCR) analysis was performed on a subset of patients to analyze the correlation between GEP data and qRT-PCR on relevant genes in patients of different cytogenetic subgroups and a variety of expression measured with GEP. The mRNA expression was determined using the average cycle threshold (Ct) in comparison to expression levels of GAPDH, using the comparative cycle threshold method. Statistical analysis Statistical analyses for disease outcome and correlation were performed with IBM SPSS 21 (IBM, Armonk, New York, USA). To assess clinical outcome, complete remission (CR; defined as less than 5% blasts in the bone marrow, with regeneration of tri-lineage hematopoiesis, plus absence of leukemic cells), probability of event-free survival (pEFS; defined as the time between diagnosis and first event including non-responders calculated as an event on day 0), and probability of overall survival (pOS; defined as the time between diagnosis and death) were analyzed. Both pEFS and pOS were estimated by the Kaplan-Meier method and differences compared using log rank tests. The cumulative incidence of nonresponse or relapse (pCIR; defined as time between diagnosis and relapse, and with non-responders included as an event on day 0) was analyzed by the Kalbfleisch and Prentice method, and compared with the Gray's test. P<0.05 was considered significant. Differential expression analyses between groups were conducted using the R-package ShirnkBayes 31 at the probe level. In contrast to commonly used two-sample tests, this test is proven to be powerful for small sample sizes. Bayesian false discovery rate (BFDR)<0.05 was considered statistically significant. Results Identification of IKZF1 deletions in pediatric AML The cohort of newly diagnosed pediatric AML patients with available peripheral blood or bone marrow samples taken at initial diagnosis included in this study has been previously described by Hollink et al. (n=293,. J.D.E. de Rooij et al. 1152 haematologica | 2015; 100(9) © F e r r a t a S t o r t i F o u n d a t i o n from which no material was available for MLPA analysis. Mutational screening of exons 4, 5 and 8 of IKZF1 was performed on the same cohort of 258 patients. This revealed no patient harboring a frameshift mutation, which has been described as damaging in BCP-ALL. We did identify 3 patients with a heterozygous point mutation, p.V382M, which was predicted as tolerated by SIFT analysis and as benign by PoyPhen analysis, and p.G158S and p.L188P, which were predicted as deleterious by SIFT analysis and probably damaging by PolyPhen analysis, but not resulting in a truncated protein. Seventy-seven of 258 patients presented with synonymous SNP rs61731355, 17/258 with synonymous SNP rs61731356, and 1/258 with non-synonymous SNP rs376657964 (p.T333A), which is in line with the presence of these SNPs in the normal population (21%, 4% and 1%, respectively). 33 In total, 10 patients carried a heterozygous IKZF1 deletion as identified by MLPA, and one additional case as identified by array-CGH Combining the data gathered from MLPA, karyotype and array CGH, we defined 11 patients with an IKZF1 deletion, 8 of which showed complete loss of one copy of chromosome 7 (monosomy 7), and 3 of which showed a focal IKZF1 deletion. As shown in Characteristics of IKZF1 deleted cases in pediatric AML No specific morphology subtypes based on the FrenchAmerican-British (FAB) classification were related to IKZF1 deletions. IKZF1-deleted samples showed none or various additional somatic mutations, most frequently IKZF1 deletions in pediatric AML haematologica | 2015; 100 © F e r r a t a S t o r t i F o u n d a t i o n activating the RAS pathway with mutations in NRAS or PTPN11 (n=4) ( 12 IKZF1 haploinsufficient AML cells display an AML-specific gene expression signature Original gene expression data are available in the Gene Expression Omnibus repository (http://www.ncbi.nlm.nih.gov/geo;accession GSE17855). Focal IKZF1 deletions are also found in mixed phenotype acute leukemia. Similarity between gene expression profiles of monosomy 7 and focal IKZF1-deleted cases Loss of IKZF1 function, a well-known tumor-inducing event in ALL, is conceivably one of the reasons for the recurrent heterozygous loss of chromosome 7, as this would include losing the IKZF1 locus on 7p12.2. We hypothesized that, if loss of IKZF1 were to be one of the driving leukemic events in monosomy 7, the gene expression signature of monosomy 7 AML cells may resemble those that only have a focal IKZF1 deletion. However, we anticipated that this similarity might be quite weak because of the heterogeneity across the samples in our cohort, both derived from somatic and germ-line genetic variation and the small sample size of monosomy 7 and IKZF1-deleted cases. Therefore, we performed genomewide differential gene expression analyses using a robust Bayesian method implemented in the R-package ShrinkBayes. 31 Probe sets were limited to those targeting unique genes or having a selectivity score of more than 0.8 (www.geneannot.com). Further statistical analyses were performed in R v. 3.0.0. (R scripts are available upon request.) We first compared gene expression profiles of monosomy 7 samples (n=8) to those of IKZF1-non-deleted samples (n=247). We detected 244 probe sets representing 198 genes differently expressed (BFDR<0.05) between these two groups ( Next, we compared the mean differences in expression of these 198 genes in monosomy 7 samples (n=8) to those in samples with IKZF1 focal deletions (n=3) ( We next examined which non-chromosome 7 genes were responsible for the similarity between monosomy 7 and focal IKZF1 deletion gene expression profiles. Genes that substantially contribute to the correlation between monosomy 7 and IKZF1-only deleted gene expression differences are up-regulated in both groups ( Gene expression profiles of the IKZF1-deleted samples demonstrate enrichment in up-regulated GATA targets HEMGN, the gene most clearly up-regulated in both monosomy 7 and IKFZ1-focal deleted samples, is transcriptionally activated by GATA1 in the myeloid cell line K562. A B C © F e r r a t a S t o r t i F o u n d a t i o n and focally deleted IKFZ1 samples are also GATA1 targets in K562. For this purpose, we used a GATA1 ChIP seq dataset in K562 cells. 39,40 GATA1 (P=0.07) and GATA3 (P=0.006) were up-regulated in the IKZF1-deleted samples (n=11) as compared to the rest (n=247) ( Discussion In the last ten years, IKZF1 has been widely studied in the context of B-cell differentiation and acute lymphoblastic leukemia, both in adults and children. There is growing evidence to indicate that IKZF1 also plays a role in various stages of myeloid differentiation. So far, the best indication that loss of IKZF1 may contribute to myeloid leukemogenesis are deletions of the short arm of chromosome 7 associated with MPN-preceded secondary AML in adults, where the commonly deleted region is mapped to the IKZF1 locus. J.D.E. de Rooij et al. 1156 haematologica | 2015; 100(9) © F e r r a t a S t o r t i F o u n d a t i o n Reduced IKZF1 gene function is a well-known recurrent event in BCP-ALL. In pediatric BCP-ALL, the overall frequency of focal IKZF1 deletions is approximately 15%, but is greatly enriched in BCR/ABL1 positive BCP-ALL at 70%-80%. In adult and pediatric BCP-ALL, in addition to focal IKZF1 deletions, monosomy 7 is a recurrent chromosomal aberration. In BCR/ABL1 positive BCP-ALLs, 16% (adult) and 13% (pediatric) of the cases of IKZF1 deletion can be attributed to monosomy 7. We hypothesized that, in pediatric AML, as presumed in BCP-ALL, an important biological determinant for leukemogenesis in monosomy 7 patients is loss of the IKZF1 gene. 34 Several genes up-regulated in both monosomy 7 and focal IKZF1-deleted AML cases are previously implicated in leukemogenesis. In a large cohort of adult BCP-ALL, comparison between focal IKZF1 deletions and IKZF1 IKZF1 deletions in pediatric AML haematologica | 2015; 100 A B C D E F © F e r r a t a S t o r t i F o u n d a t i o n wild-type cases revealed upregulation of genes involved in the cell cycle, JAK-STAT signaling and stem cell self-renewal. 21 Taken together, we find evidence for IKZF1 as a tumor suppressor gene in pediatric acute myeloid leukemia and suggest that IKZF1 deletion might be one of the driving events of monosomy 7 in various myeloid diseases. Funding JdR was funded by Kinder Oncologisch Centrum Rotterdam (KOCR). AO was funded by KIKA project #109. MF was funded by KWF project EMCR 2012-5546. The authors declare no conflict of interest. Authorship and Disclosures J.D.E. de Rooij et al. 1158 haematologica | 2015; 100(9) © F e r r a t a S t o r t i F o u n d a t i o

Recurrent abnormalities can be used for risk group stratification in pediatric AMKL: A retrospective intergroup study

Genetic abnormalities and early treatment response are the main prognostic factors in acute myeloid leukemia (AML). Acute megakaryoblastic leukemia (AMKL) is a rare subtype of AML. Deep sequencing has identified CBFA2T3/GLIS2 and NUP98/KDM5A as recurrent aberrations, occurring in similar frequencies as RBM15/MKL1 and KMT2A-rearrangements. We studied whether these cytogenetic aberrations can be used for risk-group stratification. To assess frequencies and outcome parameters of recurrent cytogenetic aberrations in AMKL, samples and clinical data of patients treated by the AIEOP, BFM-SG, COG, DCOG and the Saint Louis Hopital were collected, enabling us to screen 153 newly diagnosed pediatric AMKL cases for the aforementioned aberrations, and to study their clinical characteristics and outcome. CBFA2T3/GLIS2 was identified in 16% of the cases, RBM15/MKL1 in 12%, NUP98/KDM5A and KMT2A-rearrangements in 9% each, and monosomy 7 was identified in 6%. These aberrations were mutually exclusive. RBM15/MKL1-rearranged patients were significantly younger. No significant differences in sex, and white blood cell count were found. NUP98/KDM5A, CBFA2T3/GLIS2, KMT2A-rearranged lesions and monosomy 7 (NCK-7) independently predicted a poor outcome; compared to RBM15/MKL1-rearranged patients and to those with AMKL not carrying these molecular lesions. NCK-7-patients (n=61) showed a 4-yr pOS of 35\ub16% vs 70\ub15% in the RBM15/MKL1-other groups (n=92, p<0.0001), and 4-yr pEFS of 33\ub16% vs 62\ub15% (p=0.0013), the 4-yr cumulative incidence of relapse being 42\ub17% and 19\ub14% (p=0.003), respectively. We conclude that these genetic aberrations may be used for risk-group stratification of pediatric AMKL and for treatment tailoring

Derivation cohort baseline characteristics.

<p>Derivation cohort baseline characteristics. for mother-infant pairs with and without histological chorioamnionitis (HC; left), and with and without histological chorioamnionitis with fetal involvement (HCF; right). Abbreviations: SD = standard deviation; IQR = interquartile range; HELLP = haemolysis, elevated liver enzymes, low platelets; PPROM = preterm premature rupture of membranes.</p

Logistic regression model and clinical prediction rule for histological chorioamnionitis with fetal involvement.

<p>Logistic regression models and clinical prediction rules for prediction of histologic chorioamnionitis with fetal involvement. Abbreviations: SE = standard error; PPROM = preterm premature rupture of membranes; SGA = small for gestational age.</p

ROC curve and PPV-NPV plot of clinical prediction rule for histological chorioamnionitis (A+B) and histological chorioamnionitis with fetal involvement (C+D).

<p>Figures in PPV-NPV plots indicate inclusive (≥) cut-off values for positive test scores.</p