<p>Supplementary Figure S1 and Tables S1-3. Figure S1. Frequencies and distribution of co-operating mutations in RUNX1 mutationpositive patients. Table S1. Comparison of clinicohematological characteristics according to the biological activities of RUNX1 in MDS and CMML. Table S2. Correlation of risk of sAML transformation with RUNX1activities or co-existing gene mutations in MDS and CMML patients carrying RUNX1 mutations. Table S3. Correlation of sAML-free survival with cytogenetics or additional co-existing gene mutations in MDS and CMML patients carrying RUNX1 mutations. Supplemental methods and References</p>
The measurement of multiple behavior endpoints in zebrafish can provide informative clues within neurobehavioral field. However, multiple behavior evaluations usually require complicated and costly instrumental settings. Here, we reported a versatile setting that applied ten acrylic tanks arranging into five vertical layers and two horizontal columns to perform multiple behavior assays simultaneously, such as the novel tank diving test, mirror-biting test, social interaction, shoaling, and predator escape assay. In total, ten behavioral performance were collected in a single video, and the XY coordination of fish locomotion can be tracked by using open source software of idTracker and ImageJ. We validated our setting by examining zebrafish behavioral changes after exposure to low dose ethanol (EtOH) for 96 h. Fish were observed staying longer time at bottom of the tank, less mirror biting interest, higher freezing time, less fear in predator test, and tight shoaling behaviors which indicated the anxiogenic effect was induced by low dosage exposure of EtOH in zebrafish. In conclusion, the setting in this study provided a simple, versatile and cost-effective way to assess multiple behavioral endpoints in zebrafish with high reliability and reproducibility for the first time.
Rare earth elements (REEs) or “technology metals” were coined by the U.S. Department of Energy, a group of seventeen elements found in the Earth’s crust. These chemical elements are vital and irreplaceable to the world of technology owing to their unique physical, chemical, and light-emitting properties, all of which are beneficial in modern healthcare, telecommunication, and defense. Rare earth elements are relatively abundant in Earth’s crust, with critical qualities to the device performance. The reuse and recycling of rare earth elements through different technologies can minimize impacts on the environment; however, there is insufficient data about their biological, bioaccumulation, and health effects. The increasing usage of rare earth elements has raised concern about environmental toxicity, which may further cause harmful effects on human health. The study aims to review the toxicity analysis of these rare earth elements concerning aquatic biota, considering it to be the sensitive indicator of the environment. Based on the limited reports of REE effects, the review highlights the need for more detailed studies on the hormetic effects of REEs. Aquatic biota is a cheap, robust, and efficient platform to study REEs’ toxicity, mobility of REEs, and biomagnification in water bodies. REEs’ diverse effects on aquatic life forms have been observed due to the lack of safety limits and extensive use in the various sectors. In accordance with the available data, we have put in efforts to compile all the relevant research results in this paper related to the topic “toxicity effect of REEs on aquatic life”.
DNA methylation plays several roles in regulating neuronal proliferation, differentiation, and physiological functions. The major de novo methyltransferase, DNMT3, controls the DNA methylation pattern in neurons according to environmental stimulations and behavioral regulations. Previous studies demonstrated that knockout of Dnmt3 induced mouse anxiety; however, controversial results showed that activation of Dnmt3 causes anxiolytic behavior. Thus, an alternative animal model to clarify Dnmt3 on modulating behavior is crucial. Therefore, we aimed to establish a zebrafish (Danio rerio) model to clarify the function of dnmt3 on fish behavior by behavioral endpoint analyses. We evaluated the behaviors of the wild type, dnmt3aa, and dnmt3ab knockout (KO) fish by the novel tank, mirror biting, predator avoidance, social interaction, shoaling, circadian rhythm locomotor activity, color preference, and short-term memory tests. The results indicated that the dnmt3aa KO fish possessed abnormal exploratory behaviors and less fear response to the predator. On the other hand, dnmt3ab KO fish displayed less aggression, fear response to the predator, and interests to interact with their conspecifics, loosen shoaling formation, and dysregulated color preference index ranking. Furthermore, both knockout fishes showed higher locomotion activity during the night cycle, which is a sign of anxiety. However, changes in some neurotransmitter levels were observed in the mutant fishes. Lastly, whole-genome DNA methylation sequencing demonstrates a potential network of Dnmt3a proteins that is responsive to behavioral alterations. To sum up, the results suggested that the dnmt3aa KO or dnmt3ab KO fish display anxiety symptoms, which supported the idea that Dnmt3 modulates the function involved in emotional control, social interaction, and cognition.
A T-maze test is an experimental approach that is used in congenital research. However, the food reward-based protocol for the T-maze test in fish has low efficiency and a long training period. The aim of this study is to facilitate the T-maze conditions by using a combination of the principles of passive avoidance and a spatial memory test. In our modified T-maze settings, electric shock punishment (1–2 V, 0.3–0.5 mA) is given at the left arm, with a green cue at the right arm. Also, the depth of both arms of the T-maze was increased. The parameters measured in our T-maze design were latency, freezing time, and time spent in different areas of the T-maze. We validated the utility of our modified T-maze protocol by showing the consistent finding of memory impairment in ZnCl2−treated fish, which has been previously detected with the passive avoidance test. In addition, we also tested the spatial memory performance of leptin a (lepa) mutants which displayed an obesity phenotype. The results showed that although the learning and memory performance for lepa KO fish were similar to control fish, they displayed a higher freezing behavior during the training phase. In conclusion, we have established a modified T-maze protocol that can be used to evaluate the anxiety, learning, and memory capacity of adult zebrafish within three days, for the first time.
S172 impact on survival. In patients with BMI>25, median DFS was not reached with a 3-year DFS at 76%, while median DFS was 16.1 months with 3-year DFS at 13% for those with BMI£25 (p1⁄40.005). Median OS was not reached in T-ALL patients with BMI>25 versus 28.3 months in those with BMI£25 (3-year OS: 78% versus 41%, p1⁄4 0.04). Multivariate analyses confirmed the prognostic value of BMI (> or < to 25) in T-ALL, but only in terms of DFS (HR1⁄40.25; 95% CI: 0.05e0.87; p1⁄40.037) and not of OS. Conclusion: These findings provide further evidence that initial body size may have a potential prognostic impact in some subset of leukemia, and more specifically in T-ALL. In this AL subtype, validation in larger prospective studies is however warranted. 105 Detection of Minimal Residual Disease Using Next Generation Sequencing for Antigen Receptor Gene Rearrangements in Acute Lymphoblastic Leukemia Der-Cherng Liang, Ying-Jung Huang, Sung-Tzu Liang, Hsiao-Wen Kao, Chen-Chen Kang, Shih-Hsiang Chen, Chun-Hui Lin, Chang-Liang Lai, Chia-Hui Chang, Lee-Yung Shih Division of Pediatric Hematology-Oncology, Mackay Memorial Hospital; Chang Gung Memorial Hospital; Mackay Memorial Hospital; Chang Gung Memorial Hospital and Chang Gung University; Chang Gung Children’s Hospital Objective: Minimal residual disease (MRD) is a powerful therapeutic and prognostic indicator in acute lymphoblastic leukemia (ALL). Q-PCR for immunoglobulin and T-cell receptor (Ig/TCR) rearrangements is considered as the standard methodology. However, it is labor-intensive. We aimed to establish MRD detection technique by using next generation sequencing (NGS) for Ig/TCR targets and to compare the results of NGS-MRD with two PCRbased assays and flow cytometry. Patients and Methods: Seventyseven follow-up samples (67 B-ALL, 10 T-ALL) were collected from 55 ALL patients (48 children, 7 adults) for NGS analysis. The sequencing libraries were created with 60 ng DNA from bone marrow samples by using multiplex PCR with BIOMED-2 primers and labeled with barcodes, then sequenced with 150,000 reads on Ion Torrent PGM. MRD was identified by the matched clonal sequences derived from the diagnosis samples. MRD-negative was defined as < 0.01%. Seventy samples were examined simultaneously with QPCR-Ig/TCR, 60 with six-color flow cytometry for leukemia-associated immunophenotypes (flow-LAIP), and 30 with RQ-PCR fusion transcript assays. Correlation was made by linear regression with R square and concordance was determined by the presence of the same trait on two methods. Results: Seventy-seven samples were analyzed by NGS with a sensitivity of 2 10. A comparison between NGS and QPCR-Ig/TCR assays showed that MRD was both positive in 39 samples and both negative in 19 samples, whereas NGS-MRD (+)/QPCR-Ig/TCR (-) was found in 7 Clinical Lymphoma, Myeloma & Leukemia June 2015 samples and NGS-MRD (-)/QPCR-Ig/TCR (+) was detected in 5 samples (R1⁄4 0.8015; concordance 82.9%). NGS-MRD and flowLAIP were both positive in 35 samples and both negative in 19 samples. Six samples had NGS-MRD (+) and flow-LAIP (-), and none had NGS-MRD (-)/flow-LAIP (+) (R1⁄4 0.7299; concordance 90.0%). Both NGS-MRD and RQ-PCR fusion transcript assays showed positive results in 15 samples and negative in 8 samples whereas NGS-MRD (+)/RQ-PCR-fusion transcript (-) and NGSMRD (-)/RQ-PCR fusion transcripts (+) were observed in 1 and 6 samples, respectively. (R1⁄4 0.7458; concordance 76.7%). Conclusions: We have established a NGS-MRD detection methodology for monitoring Ig/TCR clonal targets, which exhibited a good concordance with the currently available methods. Grant support: MMH-E-102-09, CMRPG3A0723, CMRPG3B1781, CORPG3C0201 and DOH102-TD-C-111-006, Taiwan. 106 Prognostic Factors in Patients with Refractory and Relapsed Acute Lymphocytic Leukemia (ALL) Treated with Inotuzumab Ozogamicin (IO), a CD22 Monoclonal Antibody Koji Sasaki, Elias Jabbour, Susan O’Brien, Xuelin Huang, Deborah Thomas, Michael Rytting, Guillermo Garcia-Manero, Jorge Cortes, Sherry Pierce, Tapan Kadia, Hagop Kantarjian Department of Leukemia, The University of Texas MD Anderson
Animal models play important roles in investigating the pathobiology of cancer, identifying relevant pathways, and developing novel therapeutic tools. Despite rapid progress in the understanding of disease mechanisms and technological advancement in drug discovery, negative trial outcomes are the most frequent incidences during a Phase III trial. Skin cancer is a potential life-threatening disease in humans and might be medically futile when tumors metastasize. This explains the low success rate of melanoma therapy amongst other malignancies. In the past decades, a number of skin cancer models in fish that showed a parallel development to the disease in humans have provided important insights into the fundamental biology of skin cancer and future treatment methods. With the diversity and breadth of advanced molecular genetic tools available in fish biology, fish skin cancer models will continue to be refined and expanded to keep pace with the rapid development of skin cancer research. This review begins with a brief introduction of molecular characteristics of skin cancers, followed by an overview of teleost models that have been used in the last decades in melanoma research. Next, we will detail the importance of the zebrafish (Danio rerio) animal model and other emerging fish models including platyfish (Xiphophorus sp.), and medaka (Oryzias latipes) in future cutaneous malignancy studies. The last part of this review provides the recent development and genome editing applications of skin cancer models in zebrafish and the progress in small molecule screening.
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