The National Survey of Child and Adolescent Well-Being is a national probability study of children investigated for child abuse and neglect. This core study is complemented with a national ...probability study of children who have been in foster care about one year. Plans and efforts to recruit 105 county agencies, more than 6,000 children ages 0-14, and a total of nearly 25,000 respondents associated with the child are described. Parents or other permanent caregivers, foster parents for children removed from the home, and the children themselves are interviewed. For children in out-of-home placement, the caregiver from whom the child was taken is also recruited for interviews. Investigative Child Welfare Workers at the baseline, and service workers in subsequent rounds if the family is receiving services, are also interviewed about the case. In addition, an annual teacher survey is conducted for children in grades K-12. Several advances in survey methodology help to manage the process in a cost-efficient and scientifically rigorous manner. Lessons from the planning stages and from the early weeks of fieldwork are presented. The sampling and instrumentation techniques are discussed alongside other methodological issues including agency recruitment, recruitment of families, human subject protection issues, FR attrition, and data release. 2 Tables, 1 Figure, 4 References. (Original abstract - amended)
In this secondary analysis of results of the Clinical Outcomes in MEasurement-Based Treatment (COMET) trial, patient behaviors that might account for the differences observed in clinical outcomes ...were examined. Patients (N=914) diagnosed as having major depressive disorder participated in telephone interviews either monthly for six months (intervention) or at three and six months (usual care) asking about antidepressant medication-taking, use of psychotherapy or counseling, and participation in depression support groups. Physicians (N=83) in the intervention arm received monthly feedback regarding their patients' depression severity. A total of 664 (73%) patients completed the month 6 interview. The adjusted odds of current antidepressant use at six months were 85% greater (p=.01) for patients in the intervention (N=380) versus usual care (N=284) arms, according to multivariate regression analyses. More frequent measurement of depression symptoms was associated with greater medication persistence, which in turn may have mediated clinical improvements.
Abstract
Cytogenetically normal acute myeloid leukemia (CN AML) patients with a MLL PTD have a poor prognosis compared to CN AML patients with MLL WT. We previously reported that the MLL WT gene in ...MLL PTD+ CN AML is epigenetically silenced. To investigate in vivo significance of the MLL PTD in the absence of MLL WT on hematopoiesis and leukemogenesis, we generated homozygous MllPTD/PTD and hemizygous MllPTD/- mice. These mice died in utero or as neonates, respectively, precluding further study on adult hematopioesis and leukemogenesis. In the current study, we crossed MllPTD/WT mice with Mll-conditional knock-out (cKO) animals to produce MllPTD/cKO mice. Like the MllPTD/WT mice, the MllPTD/cKO mice survive to adulthood, as both models express Mll WT and PTD. As previously reported by Jude et al, (Cell Stem Cell, 2007) Cre activation in hematopoietic cells of mice carrying the Mll cKO allele resulted in an intragenic deletion in Mll (deltaN allele) that when expressed, the protein was unable to translocate to the nucleus and thereby unable to exert normal Mll function. DNA PCR confirmed the correct MllPTD/deltaN genotype was generated. MlldeltaN/deltaN mice develop bone marrow (BM) failure (marked hypocellularity) at a median of 18 days post-Cre activation. Comparatively, MllPTD/deltaN mice (n=5) survived beyond 52 weeks post-Cre activation (P<0.001), without evidence of BM failure. Also, at one year, MllPTD/deltaN animals exhibit equivalent white blood cell counts and hematocrit when compared to MllWT/WT, MllPTD/WT and MllPTD/cKO controls. To evaluate whether Mll PTD function was maintained in bone marrow, qRT-PCR analysis of the Mll transcriptional target Hoxa9 at 12 days post-Cre activation revealed a 20-fold reduction in Hoxa9 mRNA levels in MlldeltaN/deltaN animals and ∼4-5 fold increase in MllPTD/deltaN and MllPTD/cKO BM (P=0.025) compared to WT BM. MllPTD/cKO animals, with two functional copies of Mll, had nearly equivalent overexpression of HoxA9 as seen in MllPTD/deltaN mice that maintain only one functional copy of Mll, supporting our previous hypothesis that the Mll PTD acts as a gain-of-function mutation. In conclusion, these data demonstrate for the first time that Mll PTD alone can support adult hematopoiesis. In conclusion, these data demonstrate for the first time that Mll PTD alone can support adult hematopoiesis and even though Hoxa9 is upregulated, another oncogene is required for leukemogenesis. Absence of Mll WT is not sufficient to promote acute leukemogenesis in Mll PTD+ mice, but appears to be a contributory factor. We recently reported that MllPTD/WT mice crossed with Flt3ITD/WT mice, the latter of which also do not develop AML (Lee et al. Cancer Cell, 2006), generated double mutant mice that do develop AML. Leukemic blasts from these mice exhibit a reduction in Mll WT expression (Zorko et al, Blood, 2012), recapitulating what is seen in human MLL PTD+ CN AML.
Citation Format: Nicholas A. Zorko, Daniel A. Yanes, W. Courtland Lewis, Daniel L. Brook, Kelsie M. Bernot, Ronald F. Siebenaler, Susan P. Whitman, Elshafa H. Ahmed, Kathleen K. McConnell, John Nemer, Patricia Ernst, Gang Huang, Guido Marcucci, Michael A. Caligiuri. The partial tandem duplication of Mll (Mll PTD) is a gain-of-function in the absence of Mll wildtype (Mll WT) in adult mouse hematopoiesis. abstract. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3849. doi:10.1158/1538-7445.AM2013-3849
The coexpression of the MLL partial tandem duplication (PTD) and the FLT3 internal tandem duplication (ITD) mutations associate with a poor outcome in cytogenetically normal acute myeloid leukemia ...(AML). In mice, a double knock-in (dKI) of MllPTD/wt and Flt3ITD/wt mutations induces spontaneous AML with an increase in DNA methyltransferases (Dnmt1, 3a, and 3b) and global DNA methylation index, thereby recapitulating its human AML counterpart. We determined that a regulator of Dnmts, miR-29b, is downregulated in bone marrow of dKI AML mice. Bortezomib exerted a dose-dependent increase in miR-29b expression in AML blasts ex vivo, followed by decreased Dnmts, reduced proliferation, and increased apoptosis. In vivo, bortezomib was not active against dKI AML, yet liposomal-encapsulated bortezomib, as a single agent, reversed downregulation of miR-29b in vivo and induced a long-term (90-day) disease-free remission in 80% of dKI AML mice that exhibited high leukemic burden at the start of therapy, yet showed no signs of relapse at autopsy. Taken together, these data support that liposomal bortezomib, as a single agent, eradicates MllPTD/wt:Flt3ITD/wt AML in mouse and may represent a powerful and potentially curative approach to high-risk human disease.
•The MllPTD/wt:Flt3ITD/wt mouse is a relevant AML model in which the miR-29b–mediated epigenetics-kinome crosstalk is targetable by bortezomib.•An original liposomal formulation of bortezomib eradicates AML and yields curative therapy for MllPTD/wt:Flt3ITD/wt AML.
Abstract
The partial tandem duplication (PTD) of the MLL gene is a molecular defect in human myelodysplastic syndrome and acute myeloid leukemia (AML) and associates with poor patient outcomes, ...especially when other molecular prognostic markers are co-present such as the internal tandem duplication (ITD) of FLT3, i.e., FLT3-ITD. In human AML, MLL-PTD alters the epigenome via aberrant H3K4 methylation activity but is also associated with increased DNA methylation through an unknown mechanism (Whitman, et al, Blood 2008). Using a double knock-in mouse model of AML with Mll-PTD and Flt3-ITD, we analyzed the DNA methylome using a methyl binding protein (MBD2) pull down followed by next generation sequencing. Leukemic mouse bone marrow (BM, n=8) exhibited a 1.7 fold increase in global DNA methylation compared to non-leukemic controls (n=9, P=0.017). Consistent with this finding, DNA methyltransferases (Dmnt) 1, 3a, and 3b were overexpressed (1.5 fold P=0.03, 2.3 fold P=0.01, and 5.3 fold P=0.03, respectively). We previously showed that the non-coding microRNA 29b (miR-29b) expression directly downmodulates DNMT3a and 3b, but indirectly inhibits DNMT1 via miR-29b-driven SP1 suppression. In the murine Mll-PTD/Flt3-ITD AML, the precursor to the mature miR-29b (pri-miR-29b-2) was downregulated in leukemic BM (0.7-fold, P<0.001), whereas Sp1 mRNA was upregulated compared to wildtype (1.6-fold P=0.01). Bortezomib is a proteasome inhibitor that also downregulates Dnmt's (Liu, et al, Blood, 2008) by increasing the expression of miR-29b (Liu, et al, Cancer Cell, 2010). In vitro, bortezomib induced a dose-dependent reduction in proliferation of Mll-PTD/Flt3-ITD leukemic blasts (MTS assay, EC50 23.83nM) and apoptosis. Prior to cell death in bortezomib-treated cells, expression of pri-miR-29b-2 increased by 1.7 fold over vehicle treated cells (P=0.01) while Dnmt3b mRNA (P=0.001) and protein levels decreased to half of the vehicle treated level. Targeting of this pathway in vivo using liposomal-bortezomib (1mg/kg IV twice per week for 2 weeks followed by 2mg/kg IV twice per week for 2 weeks) significantly increased survival of AML transplanted mice, compared with vehicle or free-bortezomib with 80% alive 90 days post-transplant (P<0.001). At day 90, treated mice had normal spleen weights (mean 86mg) and absence of leukemic blasts in BM, liver, or spleen compared to vehicle-treated mice that showed significant blast infiltration (mean spleen weight 396mg, P<0.001). Taken together, these data support the miR-29b/SP1/DNMT pathway of epigenetic modulation as active in Mll-PTD/Flt3-ITD AML, and indicate bortezomib as a valuable treatment option to be explored in patients with MLL-PTD/FLT3-ITD-associated AML.
Citation Format: Kelsie M. Bernot, John S. Nemer, Ramasamy Santhanam, Shujun Liu, Gabriel G. Marcucci, Nicholas A. Zorko, Susan P. Whitman, Pearlly Yan, David Frankhouser, Ralf Bundschuh, Mengzi Zhang, Ronald F. Siebenaler, Elshafa H. Ahmed, Kathleen K. McConnell, Maura Munoz, Daniel L. Brook, Adrienne M. Dorrance, Katy E. Dickerson, Xiaoli Zhang, Jianying Zhang, David Jarjoura, Robert Lee, William Blum, Michael A. Caligiuri, Guido Marcucci. Targeting the miR29b/Sp1/Dnmt pathway for curative therapy in Mll-PTD/Flt3-ITD acute myeloid leukemia. abstract. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3249. doi:10.1158/1538-7445.AM2013-3249
Rapid identification of metabolites of compound X using data dependent scan function of a quadrupole ion trap mass spectrometer and semi-automated metabolite identification software is described. ...Compound X is metabolized via monooxygenation and desmethylation. LC-ESI-MS spectra obtained, following incubations of Compound X with microsomes in the presence and absence of chemical inhibitors specific for CYP1A2, CYP3A4, CYP2D6, CYP2C9 and CYP2E1, were processed using semi-automated metabolite identification software to extract information and to identify the cytochrome P450 enzymes responsible for metabolite formation. Chemical inhibition data suggests that the primary cytochrome P450 (CYP450) isozyme responsible for the metabolism of compound X is CYP3A4 with a minor contribution from both CYP2D6 and CYP2E1. Additionally, neither CYP2C9 nor CYP1A2 appears to contribute to the metabolism of compound X.
Travel Agents and Their Survival Thomas Siebenaler, C.; Groves, David L.
Journal of human resources in hospitality & tourism,
20/1/1/, Letnik:
1, Številka:
1
Journal Article
Recenzirano
The travel agency industry depends upon commissions for a majority of its revenue. When these commissions decrease or stop the industry must change the means in which revenue is made. The Internet ...has caused a significant reduction in airline ticket sales and therefore has caused travel agencies to change. In addition travel agencies have reduced commissions paid to airlines thus causing more changes. There were four categories of travel agencies discussed in the article along with their adaptation styles. These types include (1) the independent operator of a small travel agency with very limited resources (2) the normal agency with 2 or 3 agents which does a substantial business (3) the larger agency that is high-tech and high touch and depends on automation and the Web to some extent (innovators) (4) agencies that are Web based with very little automation otherwise and who were technologically adaptable and saw opportunity in the Web. The study examines how agencies have adapted and prospered through this transitional period.
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