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Many people who lived in affected areas by the Great East Japan Earthquake and Tsunami have been forced to live separately after the event. In some tsunami-affected areas, reconstruction of houses has been restricted since the disaster happened until reconstruction projects for safety finish because sea walls were destroyed and high risk of a future tsunami remains. These areas were designated as two separated zones; prohibited area of housing reconstruction and land leveling area. Survivors who lived in either zone can also choose to move out from the area they lived and to reconstruct their house by themselves. Therefore, community recovery plan makes an impact for affected people to decide where they reconstruct, consequently, it is considered to make an effect on community recovery process. For this reason, this chapter attempts to analyze problems of community recovery plan and characteristics of residents who have a will to reconstruct their house in tsunami-affected area in case of Minami-Kesennuma, a devastated area by the tsunami. From the analysis, lessons of community recovery is emphasized as following; early concept-making and consensus-building for recovery, housing reconstruction support in terms of re-building community and resilience building in the recovery process by involvement of multi-stakeholders.

Peromyscus leucopus linville (PLL) is a mouse strain of the species P. leucopus commonly referred to as ‘white-footed’ mice. These animals share many physical features with the popular laboratory mice Mus musculus (MM) and are the most populous rodent species in the north-eastern United States (Joyner et al. 1998). P. leucopus have been used for studies of social interactions and stress physiology (Southwick 1964), as well as for the investigation of photoperiodism and the action of melatonin on the regulation of hormone secretion and reproductive physiology (Glass & Lynch 1982; Glass & Knotts 1987; Glass & Dolan 1988; Weaver et al. 1990). These animals came to public attention as the primary reservoir host for Borrelia burgdorferi sensu stricto, the aetiological agent of lyme disease (Magnarelli et al. 1988, 1994; Hofmeister & Childs 1995; Hofmeister et al. 1999), and they may also be hosts for species of hantavirus (Hjelle et al. 1995; Lyubsky et al. 1996; Morzunov et al. 1998). More recent studies have focused on behavioural and reproductive physiology of P. leucopus mice using different breeding practices under long-term captivity, with data showing that adaptation can be rapid, affecting reproductive patterns and behaviour, even under breeding protocols designed to minimize the rate of genetic change due to random drift and inadvertent selection (Malo et al. 2010; Lacy et al. 2013). Peromyscus leucopus have long lifespans that distinguish them from MM mice and from sister species within the genus Peromyscus (Sacher & Hart 1978; Steger et al. 1980; Cohen et al. 1987; Burger & Gochfeld 1992). Peromyscus show extended reproductive lifespan (Steger et al. 1980; Burger & Gochfeld 1992), accelerated wound repair (Cohen et al. 1987), reduced cellular reactive oxygen species (ROS) (Labinskyy et al. 2009; Shi et al. 2013) and increased vascular resistance to oxidative stress and inflammatory damages (Labinskyy et al. 2009). Despite their broad utilization in biomedical research, P. leucopus mice have not been well characterized. For example, haematological parameters are not fully available, because previous studies only provided partial measurements: one study reported haematocrit and total and differential WBC counts in a group of young P. leucopus mice, and another immunological and haematological values in pups born to normal dams or dams exposed to aroclor 1254 (Wu et al. 1999a,b). In the current work, we performed haematological measurements in PLL mice with reference to MM mice using C57BL/6 (B6) as representatives. We also attempted to test the effects of ageing and antioxidant N-acetylcysteine (NAC) treatment on haematological changes in P. leucopus animals. Here, we report that P. leucopus have lower platelet counts, fewer megakaryocytes and more monocytes than do MMB6 mice, with values closely resembling those of humans, suggesting that P. leucopus mice could potentially be a good model of some aspects of human haematology.

Subcutaneous panniculitis-like T-cell lymphoma (SPTCL) is an uncommon type of peripheral T-cell lymphoma (PTCL) usually presenting with erythematous, subcutaneous nodules and fever [1]. It is consi...

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired bone marrow disorder caused by expansion of a clone of hematopoietic cells lacking glycosylphosphatidylinositol (GPI)-anchored membrane proteins. Multiple lines of evidence suggest immune attack on normal hematopoietic stem cells provides a selective growth advantage to PNH clones. Recently, frequent loss of HLA alleles associated with copy number-neutral loss of heterozygosity in chromosome 6p (CN-6pLOH) in aplastic anemia (AA) patients was reported, suggesting that AA hematopoiesis 'escaped' from immune attack by loss of HLA alleles. We report here the first case of CN-6pLOH in a Japanese PNH patient only in GPI-anchored protein positive (59%) granulocytes, but not in GPI-anchored protein negative (41%) granulocytes. CN-6pLOH resulted in loss of the alleles A*02:06-DRB1*15:01-DQB1*06:02, which have been reported to be dominant in Japanese PNH patients. Our patient had maintained nearly normal blood count for several years. Our case supports the hypothesis that a hostile immune environment drives selection of resistant hematopoietic cell clones and indicates that clonal evolution may occur also in normal phenotype (non-PNH) cells in some cases.

This chapter deals with three cases of knowledge-based DRR activities conducted by SEEDS Asia, a Japan-based NGO, in Myanmar, Vietnam, and Japan. From the case of Myanmar, it is clarified that even short-term DRR training initiated by NGOs can enhance capacity on DRR. From the case of Vietnam, it is proved that the working group consisted of teachers and educational administrative officers to develop DRR education programs and materials is effective for raising motivation and ownership, and NGOs can play vital role to organize the working group and to advocate to educational administrative plan for sustainability. In the case of Japan, the importance of DRR education with community was shown and it was indicated NGOs could be a bridge between school and local resources. Furthermore, it was found that DRR education network with community functioned not only for disaster preparedness but also for disaster response and recovery based on experiences of Kesennuma City. At the last, it is stressed through the three cases that one of the key issues of education is working with local government, and have trust with the school teachers and educational administrative staff.

Sound‐pressure level (SPL) fluctuation caused by air conditioners was measured by use of pure tones as the sound source in a gymnasium. When the air conditioner was turned on, the fluctuation of the SPL was more than 2 dB at 2 kHz, 5 dB at 4 kHz, 6 dB at 8 kHz, and 10 dB at 16 kHz. The cumulative density of mean‐squared amplitude corresponds well with the gamma distribution. From this statistical analysis, it is considered that the SPL change is caused by the impulse response which is represented by the direct sound (regular) and changing delay time of reflection sound (irregular). The cumulative density of mean‐squared amplitude for this model by using the pure tone also indicated the gamma distribution. It is found that the frequency characteristic and the cumulative density of the SPL change have the same tendencies as the method of modulation for the delay time of reflection sound.

In the aftermath of the Great East Japan Earthquake and Tsunami (EJET), which caused enormous damages, more than 53,000 temporary housing units were provided by the government based on the law. It is mentioned in the first half of this chapter that care enhancement activities and promoting the development of a residents’ association are vital in temporary housing due to features of residents and habitation in temporary housing with low quality from the results of surveys conducted by Ministry of Health, Labour and Welfare. The last half of this chapter analyzes key lessons of community development in temporary housing in the case of Kesennuma City. The results indicate that creating communication opportunities to share problems and establish relationships with the municipality and specialists for solving the problems is crucial to community development. In addition, needs in the fields of medical care, health care, welfare, and intergenerational exchange increased in temporary housing sites.

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired GPI-anchor deficiency caused by the somatic mutation of PIGA gene in a hematopoietic stem cell (PIGA-PNH). Loss of function of PIGA causes loss of GPI anchored proteins including complement regulatory proteins. Main symptoms are hemolytic anemia and venous thrombosis caused by the uncontrolled complement activation, and bone marrow failure. For X-linked PIGA, one hit of somatic mutation causes GPI-deficiency. All other genes involved in GPI-anchor biosynthetic pathway are autosomal and hits to two alleles are required to cause GPI-deficiency. In 2013, a PNH patient caused by the PIGT mutations was reported. This patient had a splice site mutation in one germ line allele and a somatic 8Mb deletion including entire PIGT. Recently, we found a Japanese patient who has a nonsense mutation in a germ line allele and a somatic 18Mb deletion in chromosome 20q including PIGT. Here we compare two patients and report that these cases of PNH are atypical in two points. One is that PIGT-PNH cases had unusual symptoms similar to auto inflammatory syndrome. Both had long lasting chronic urticaria and joint pain. Additionally, the Japanese case had recurrent aseptic meningitis and the German patient had ulcerative colitis. It should be noted that these symptoms had lasted over ten years before hemolytic attacks occurred and diagnosis as PNH was made. Urticaria, joint pain and recurrent meningitis were controllable by eculizumab, anti-C5 antibody, showing involvement of complement activation. These inflammatory symptoms are specific to PIGT-PNH and are not seen in PIGA-PNH. PIGT is involved in transfer of GPI to proteins and its defect causes accumulation of free GPI, non-protein linked GPI. In fact, FACS analysis using the antibody which recognizes free GPI revealed that GPI negative granulocytes, monocytes, and B cells from the PIGT-PNH patient, but not from PIGA-PNH patients, highly expressed free GPI. We speculate that this free GPI together with complement activation cause inflammasome activation and are establishing assay system. The other atypical nature is a mechanism of clonal expansion. In typical PNH, autoimmunity against hematopoietic stem cells causing bone marrow failure is thought to positively select GPI-deficient cells. In contrast, the somatically deleted 18Mb included a region commonly deleted in some patients with myeloproliferative noeplasms. This region contains several maternally imprinted genes, defective expression of which is likely causally related to clonal expansion in atypical PNH cases. Thus, the symptoms and mechanism of clonal expansion of PIGT-PNH are quite different from those of PIGA-PNH, so that PIGT-PNH should be categorized as an atypical PNH. Screening of PIGT-PNH among the PNH patients by detecting free GPI using FACS analysis should be feasible. Disclosures Ueda: Alexion Pharmaceuticals: Honoraria, Research Funding. Nishimura:Alexion Pharma: Honoraria, Research Funding. Kanakura:Bristol Myers: Research Funding; Toyama Chemical: Research Funding; Kyowa Hakko Kirin: Research Funding; Eisai: Research Funding; Shionogi: Research Funding; Chugai Pharmaceutical: Research Funding; Fujimotoseiyaku: Research Funding; Pfizer: Research Funding; Nippon Shinyaku: Research Funding; Alexionpharma: Research Funding; Astellas: Research Funding. Kinoshita:Alexion Pharma: Honoraria.

Introduction: Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired stem cell disorder with the expansion of PIGA mutant clone(s), which is deficient in GPI-anchored proteins including CD55 and CD59. The lack of CD55 and CD59 renders PNH-type red blood cells (RBC) susceptible to complement attacks, resulting in intravascular hemolysis. Classic PNH manifests 3 major symptoms: anemia, bone marrow failure, and thrombosis. Small populations of PNH-type cells ( Methods: Patients diagnosed with PNH, AA, MDS or indistinguishable BMF were prospectively recruited to the study since 2011 in Japan. A high-resolution FCM assay was established to precisely detect a small population of PNH-type granulocytes (with FLAER) and RBCs (with anti-CD55 and CD59 antibodies) ≤ 0.01% of the total granulocyte or RBC population based on the Kanazawa method (Blood 2006 107:1308-1314). Six university laboratories across Japan were designated as regional analyzing centers and measured the percentages of PNH-type cells in the study population, as well as collected clinical and laboratory data. Periodic blind cross validation tests using a positive control sample containing 0.01% PNH-type cells and a negative control sample were performed to minimize inter-laboratory variations. Results: As of July 2016, 2,849 patients were enrolled in the study and 2,734 patients were analyzed. Nine hundred twelve patients (33.4%) were positive for PNH-type cells (≥ 0.005% PNH-type erythrocytes and/or ≥ 0.003% PNH-type granulocytes) and 238 (8.7%) patients had more than 1% of PNH-type cells. PNH-type cells were positive in 90/90 PNH (100%), 512/982 AA (52.1%), 132/822 MDS (16.1%), and 141/512 indistinguishable BMF (27.5%) patients. Among the MDS patients, PNH-type cells were positive in approximately 20% of patients with RCUD, RCMD, MDS-U, or 5q- syndrome, but not in any patients with RARS, RAEB-1, and RAEB-2 (Fig. 1). The serum LDH level increased in proportion to the PNH clone size of RBCs, and 63.3% of the patients possessing ≥1.0% RBCs showed LDH levels more than 1.5 times the upper limit of normal. Of 171 patients who completed submission of 3-year follow-up data, BMF patients with PNH-type cells showed a better response rate [CR+PR, 99/107 (92.5%)] to IST compared to those without PNH-type cells [44/64 (68.8%)] (P Conclusions: These interim analyses of the OPTIMA study demonstrate that our high-resolution FCM is reliable in detecting small populations of PNH-type cells and produces consistent results among different laboratories. The presence of PNH-type cells exclusively in patients with AA and low-risk MDS suggests a link between benign pathophysiology of BMF and an increase in the number of PNH-type cells. The better response of PNH-type cell-positive BMF to IST compared to BMF without PNH-type cells was consistent with previous reports. Our data, for the first time, prospectively confirms the significance of small populations of PNH cells in BMF patients in Japan and warrants further worldwide, prospective studies on non-Japanese patients with BMF. Disclosures Ueda: Alexion Pharmaceuticals: Honoraria, Research Funding. Nishimura:Alexion Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Hosokawa:Aplastic Anemia and MDS International Foundation: Research Funding. Yonemura:Alexion Pharmaceuticals: Research Funding. Obara:Alexion Pharmaceuticals: Honoraria, Research Funding. Shichishima:Alexion Pharmaceuticals, Inc. Japan: Honoraria. Ninomiya:Alexion Pharmaceuticals: Honoraria. Kawaguchi:Alexion Pharmaceuticals: Honoraria. Kanakura:Fujimotoseiyaku: Research Funding; Toyama Chemical: Research Funding; Bristol - Myers: Research Funding; Nippon Shinyaku: Research Funding; Astellas: Research Funding; Eisai: Research Funding; Pfizer: Research Funding; Chugai Pharmaceutical: Research Funding; Shionogi: Research Funding; Kyowa Hakko Kirin: Research Funding; Alexion Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Nakao:Alexion Pharmaceuticals: Honoraria, Research Funding.

Alter, Durand and Froment introduced the total-variation (TV) minimization approach to the artifact-free JPEG decoding, which is referred to as the ADF decoding method [1]. They formulated the decoding problem as the constrained TV restoration problem, in which the TV seminorm of its restored color image is minimized under the constraint that each DCT coefficient of the restored color image should be in the quantization interval of its corresponding DCT coefficient of the JPEG-compressed data. This paper proposes a new restoration approach to the JPEG decoding. Instead of the TV regularization, our new JPEG-decoding method employs a shrinkage operation in the redundant DCT domain, to mitigate degradations caused by the JPEG coding. Our new method not only can selectively suppress ringing artifacts near color edges, but also can efficiently eliminate blocking artifacts in originally smoothly-varying image regions, where the blocking artifacts are very noticeable. Through decoding simulations, we experimentally show that our new decoding method can reduce JPEG-coding artifacts more effectively than the ADF decoding method.

Background: Paroxysmal Nocturnal Hemoglobinurea (PNH) is a rare stem cell disease caused by the expansion of PIGA mutated clone(s). PNH-type cells are deficient in the expression of GPI-anchored proteins including DAF and CD59, which protect red blood cells (RBC) from complement-mediated intravascular hemolysis. Eculizumab (Soliris®, Alexion Pharmaceuticals) is a humanized monoclonal antibody against C5 which efficiently inhibits hemolysis by blocking the terminal complement cascade. Eculizumab dramatically ameliorates several clinical symptoms, and improves the prognosis in PNH patients. However, among 345 Japanese PNH patients who were treated with eculizumab, 11 patients showed poor response. All the poor responders had a single missense C5 heterozygous mutation, c.2654G>A, which predicts the polymorphism p.Arg885His (Nishimura et al, NEJM. 2014 13;370(7):632-9). Two of those patients have already passed away due to severe complications related to PNH, and the rest of them are still suffered from various clinical symptoms including hemolytic episodes and RBC transfusion. In these circumstances, multiple new anti-complement drugs are under development in Japan. Coversin (Volution Immuno Pharmaceuticals) is a recombinant protein (16,740 Da) derived from a secreted protein in the saliva of the Ornithodoros moubata tick, and it blocks complement-mediated hemolysis at C5 level. In this study, we examined this new anti-complement agent to a PNH patient with C5 polymorphism c.2654G>A, as well as those without the polymorphism. Materials: Peripheral blood samples were collected from a poor responder to eculizumab and hemolytic PNH patients with written informed consent as approved by the Institutional Review Board of Osaka University Hospital. In vitro hemolytic assay: RBC from ABO-matched PNH patients off eculizumab treatment were washed 3 times in saline, and subsequently incubated with Mg2+ supplemented serum of the poor responder in the presence or absence of an anti-complement agent. Alternative pathway was activated by adding HCl (22:1 of 0.4M HCl) to the serum. Heat-inactivated (56°C for 30min) serum was used as a negative control. After a 24-hour incubation at 37°C, hemolysis was quantified by measuring the optical density at 405nm (OD405). The hemolytic activity was normalized against maximum hemolysis as induced by HCl (100%) and minimum hemolysis with inactivated acidified serum (0%). Results: A 41-year-old male with fatigue was diagnosed as aplastic anemia with PNH in 2008, and cyclosporine (CyA) was initiated at the dose of 150mg/day. The PNH clone expanded from 30.6% to 70.2% in granulocytes from 2008 to 2011 with elevated LDH (700 U/L) and the patient was referred to our hospital to undergo eculizumab treatment. CyA was reduced to 100mg/day and eculizumab was initiated in May 2012. Eculizumab treatment did not change the serum LDH level without any improvement of the symptoms: fatigue, abdominal pain, and periodical hemoglobinurea. A heterozygous mutation c.2654G>A was identified as the cause of the failure to eculizumab treatment, and he is still suffered from continuous intravascular hemolysis (LDH > 1400 U/L) with periodical acute hemolytic episodes, requiring frequent RBC transfusion. In the hemolytic assay, Coversin completely blocked hemolysis at the concentration of 10ug/ml, similar to the effective inhibition with hemolytic PNH patients without the polymorphism. Discussion: Eculizumab has dramatically improved the quality-of-life in the majority of the PNH patients by blocking intravascular hemolysis, but there are still some concerns; poor response due to C5 polymorphism, C3b deposition on the RBC, high cost and burden for scheduled infusion. Blocking the complement cascade at C5 level has shown to be relatively safe if meningococcal vaccination is properly performed, but still an extravascular hemolysis remains problematic at least in some cases. Inhibiting C3 amplification would resolve both intra and extravascular hemolysis, but susceptibility to infections remains a major concern. Our study showed that Coversin efficiently blocked in vitro hemolysis in the eculizumab resistant patient with C5 heterozygous mutation, c.2654G>A. Coversin might be a therapeutic option for the population of C5 polymorphism c.2654G>A in PNH patients. Our results warrant further investigation to explore new anti-complement agents for hemolytic PNH patients. Disclosures Ueda: Alexion Pharma: Research Funding. Osato:Alexion Pharma: Research Funding. Weston-Davies:Volution Immuno Pharmaceuticals (UK) Ltd: Employment, Equity Ownership. Nunn:Volution Immuno Pharmaceuticals: Employment, Equity Ownership. Hayashi:Alexion Pharma: Research Funding. Nishimura:Alexion Pharma: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Kanakura:Alexion Pharma: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

Background: Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hematopoietic stem cell disorder caused by the clonal expansion of the phosphatidylinositol glycan class A (PIGA) mutant hematopoietic stem cells, which results in a deficiency in glycosylphosphatidylinositol-anchored proteins (GPI-APs). Through the high-resolution flow cytometry-based method, GPI-APs deficient blood cells (i.e., PNH-type cells) are often detectable in patients with bone marrow failure syndromes (BMF), such as aplastic anemia (AA) and low-risk types of myelodysplastic syndromes (MDS). Sugimori et al reported that when BMF patients possessed increased PNH-type cells, the patients had a good prognosis and showed a high response rate to immunosuppressive therapies, suggesting that detection of PNH-type cells is potentially useful in determining an optimal treatment for BMF patients. Thus, we conducted a nationwide, multi-center prospective observational investigation, the OPTIMA study. Methods: From July 2011, we start recruiting the patients with BMF that were diagnosed at various hematology clinics throughout Japan to the OPTIMA study. The primary endpoint of this study was to determine the prevalence of BMF patients with PNH-type cells and to clarify the clinical significance of the presence and quantitative changes of these cells with regard to the clinical features. Six different university laboratories were assigned as regional analyzing centers. The percentage of PNH-type cells was measured by the high-resolution flow cytometry-based method, originally established in Kanazawa University. At six individual laboratories, cross validations were conducted twice a year to minimize the inter-laboratory variations in the detection sensitivities, cutoff values, etc. The liquid FLAER method (≥0.003%) and cocktail method (≥0.005%) with CD55 and CD59 antibodies were used for the detection of PNH-type granulocytes and erythrocytes, respectively. Results Between July 2011 and May 2015, a total of 2328 patients were enrolled to this study, and we analyzed 2212 patients who were eligible for the interim analysis. Of these patients, 74 (3.3%) were diagnosed with PNH, 690 (31.2%) with AA, 592 (26.8%) with MDS, and 856 (38.7%) with undiagnosed BMF. Using high-resolution flow cytometry-based method, 755 (34.1%; 95.9% in PNH, 52.8% in AA, 18.2% in MDS, and 24.8% in undiagnosed BMT) patients had ≥0.005% PNH-type erythrocytes and ≥0.003% PNH-type granulocytes. Overall, 181(8.2%) patients had ≥1% of both PNH-type erythrocytes and granulocytes; the prevalence in each disease subset was 68/74 (91.9%) in PNH, 67/690 (9.7%) in AA, 22/592 (3.7%) in MDS, and 24/856 (2.8%) in undiagnosed BMF. Regarding FAB and WHO classifications of MDS subtype, no patients with RARS (0/22), RAEB-1 (0/37) or RAEB-2 (0/23) had PNH-type cells. In contrast, 20.4% (56/275) patients with RCMD, 18.3% (26/153) patients with RCUD and 50% (2/4) patients with del (5q) MDS possessed increased PNH-type cells. Blood samples from 75 (65 with and 10 without PNH-type cells) patients were analyzed three years after the first examination. Of 65 PNH (+) patients, PNH-type cells disappeared in 4 (6.2%), while the percentage remained stable in 61 (93.8%). All of the 10 PNH (-) at the enrollment were also negative for PNH-type cells in 3 years. Conclusions: A high-resolution flow cytometry-based method that enables the detection of minimal PNH-type cells below 0.01% was successfully transferred from Kanazawa University to other laboratories in Japan. Our interim analysis confirmed previous findings that PNH-type cells were detectable in patients with 52.8% of AA and 18.2% of MDS patients. Regarding FAB and WHO classifications of MDS subtype, PNH-type cells were not detected in any of MDS RARS, RAEB-1 or RAEB-2 patients. Further analysis are required to determine the clinical significance of the minimal level of PNH-type cells as well as chronological changes in the PNH-type cell percentage, especially in terms of their relation to response to immunosuppressive therapy. Disclosures Ninomiya: Alexion Pharmaceuticals: Honoraria. Ando:Eisai Co., Ltd.: Honoraria, Research Funding. Yonemura:1. Chugai Pharma, 2. Alexion Pharma, 3. Japan Blood Products Organization, 4. OHARA Pharma: Research Funding. Kawaguchi:Alexion Pharmaceuticals: Honoraria. Ueda:Alexion Pharma: Research Funding. Nishimura:Alexion Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

Digital television is transforming both broadcasting and, as a result of convergence, the larger world of communications. The impending analogue switch-off will have a major impact on households all over the developed world. Digital Broadcasting considers the effects of digital television on the availability, price and nature of broadcast services in the Americas, Europe and Japan. It shows how this depends upon what platforms – cable, satellite, fixed or wireless broadband – countries have available for use and also upon government policies and regulatory interventions.

Background Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal stem cell disorder, characterized by the complement-mediated intravascular hemolysis. Eculizumab is a humanized monoclonal antibody that binds to the human C5 protein, thereby inhibiting terminal complement-mediated hemolysis. PNH patients treated with eculizumab responded with a significant reduction in hemolysis and improved in quality of life (QOL). Recent report shows that PNH-type cells were under the oxidative stress due to elevated levels of intracellular reactive oxygen species (ROS) and/or decreased antioxidant status. Fermented papaya preparation (FPP), a dietary nutriceutical derived from carica papaya, has been shown to modulate oxidative stress damage parameters in vitro and in vivo. To clarify the pathogenetic link between oxidative stress and hemolysis in PNH, the effects of each inhibitor, eculizumab and FPP, in Japanese patients with PNH were investigated independently. Methods Peripheral blood and sera were obtained from patients with PNH and healthy individuals after the obtaining informed consent. The study protocol was approved by the IRB at Osaka University Graduate School of Medicine. For ROS assay, RBCs were measured using 2’-7’-dichlorofluorescin-diacetate. The oxidative stress index (OSI) was derived from the percent ratio of the Reactive Oxygen Metabolites-derived compounds test (Diacron) and the Biological Antioxidant Potential test (Diacron) using a JCA-BM1650 analyzer (JEOL). LDH, biomarker of hemolysis, was measured by routine tests. Eculizumab was administrated according to standard schedule. FPP was taken orally 18g/day for 3 month. Health-related fatigue and QOL were also assessed using the FACIT-fatigue (Version 4A) and EORTC QLQ-C30 (Version 3) instruments. Results PNH patients without eculizumab treatment (pts w/o ecu) showed significantly higher levels in ROS compered to healthy individuals ([mean±SD] 1.13±0.18 vs. 0.84±0.10, p=0.004, n=8). Among these patients, CD59-negative RBCs showed a significant increase in ROS compared to GPI-positive RBCs in corresponding patients (1.23±0.26 vs. 1.13±0.27, p=0.03, n=6). Similarly, the OSI level was significantly higher in pts w/o ecu than healthy individuals (1.32±0.37 vs. 0.97±0.10, p=0.022, n=9) and oxidation activity in sera was significantly higher in pts w/o ecu than healthy volunteers (361.6±91.5 vs. 285.0±31.8, p=0.039, n=9), while antioxidant activity in sera was not significantly different between these 2 groups ([2440.6±191.5 vs. 2601.5±186.6, p=0.09, n=9). Interestingly, there was no significant difference in OSI level between PNH patients on eculizumab (n=8) and healthy individuals (0.84±0.10 vs. 1.07±0.45, p=0.55). Two patients who newly started eculizumab treatment showed improvement in LDH along with ROS generation ([LDH(IU/l)] Pt.I:6082-906, Pt.II:618-211; [ROS] Pt.I:1.57-1.06, Pt.II:1.50-1.40). FPP treatment (n=2) showed little effect on LDH, while it decreased ROS (Pt.I:1.72-1.47, Pt.II:1.16-1.04) and improved the scores in FACIT-fatigue and EORTC QLQ-C30. In addition, those 2 patients showed increase in ROS after discontinuation of FPP (Pt.I:1.47-2.04, Pt.II:1.04-1.19). Conclusion The RBCs and sera derived from Japanese patients with PNH were highly susceptible to oxidative stress compared to healthy individuals. Eculizumab was effective in controlling the oxidative stress, in addition to the hemolysis, suggesting that elevated oxidative stress in PNH was mainly due to the complement-mediated hemolysis. Since FPP showed little effect on hemolysis but had a potential to ameliorate the oxidative stress and improved QOL, FPP could alleviate symptoms associated with oxidative stress and may contribute to the therapeutic option for supportive therapy in PNH. Disclosures Osato: Alexion Pharma: Research Funding. Nishimura:Alexion Pharma: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Hayashi:Alexion Pharma: Research Funding. Ueda:Alexion Pharma: Research Funding. Kanakura:Alexion Pharma: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

The pathophysiology of human aplastic anemia (AA) is immune-mediated destruction of bone marrow stem and progenitor cells. Most patients respond to immunosuppressive therapies (IST), which markedly improved survival in this disease. However, a minority of patients undergoes transformation to malignant hematologic disease, myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML), usually accompanied by the cytogenetic abnormality monosomy 7. Clonal evolution in AA confers a poor prognosis in the clinic but is an opportunity to assess early events in oncogenesis in the setting of inflammation and tissue regeneration. We previously reported that low mean telomere length of leukocytes at the time of diagnosis of AA (Scheinberg et al., JAMA 2010) was associated with increased risk of progression to MDS. In the current study, we directly compared acquired mutations in candidate genes and chromosomal instability, as measured by telomere length, in a cohort of AA patients that had progressed to MDS. Thirteen AA patients who developed monosomy 7 were compared with 30 AA patients who had received similar treatments but did not progress to MDS. Leukocytes telomere content was measured by qPCR in samples obtained at different time points from the diagnosis of AA and chromosome-specific telomere length was assessed by single telomere length assay (STELA) for Xp, Yp, 12q, and 17p. In the AA patients who had evolved to MDS and AML, analysis of acquired mutations in myeloid-specific genes was performed by comparison with control “germline” DNA from purified CD3 lymphocytes by exome sequencing. Cells from AA patients with clonal evolution showed marked progressive telomere attrition, 419 bp/year during the period preceding development of monosomy 7. Telomere attrition was progressive from the time of diagnosis of AA. By STELA, accumulation of very short telomere fragments was apparent at 6 months after IST. In contrast, for the AA control group, patients whose disease was stable, telomere attrition was not accelerated in serial qPCR determinations, nor was there increased accumulation of short telomere fragments by STELA. A similar pattern of increased telomere attrition was reproduced in vitro by cultivation of bone marrow cells obtained six months after IST in all AA patients who developed MDS, while none of the AA control bone marrow cultured cells developed shorter telomeres. We examined bone marrow myeloid cells at the time of monosomy 7 for acquired mutations in 125 candidate genes reported to be recurrently mutated in AML and MDS. Exome sequencing was performed using Agilent SureSelect Target Enrichment System. The raw reads were mapped to UCSC Human Genome hg19 by BWA software with default setting. With an average of 111-fold coverage on selected exon regions, somatic mutations were identified between paired samples using SAMtools and Shimmer software for SNP detection. Acquired mutations in myeloid cells were found in two cases. One patient had a heterozygous mutation in DNMT3A (K829T) present since diagnosis of AA. The other patient also had a heterozygous mutation in DNMT3A (P904S) as well as mutations in DOTL1, ASXL1, SETBP1, and STAT3. All these mutations were identifiable after IST as neutrophils recovered. Despite the presence of multiple mutations, this patient had shown a good hematologic response to IST; evolution was manifest as recurrent pancytopenia and stable marrow myeloblasts at about 5% for over 2 years after first detection of monosomy 7. The remaining 11 patients, all of whom lacked candidate gene mutations, had progressive increase in bone marrow myeloblast numbers; the only other three survivors in this cohort had received hematopoietic stem cell transplant. In conclusion, telomere shortening rather than accumulation of point mutations in hematopoietic cells preceded aneuploidy and malignant transformation at an early stage of oncogenesis in this group of patients. These results from AA may be generalizable to other cancers arising in the setting of inflammation and tissue regeneration in other organs. Identification of critically short telomeres before the development of cytogenetic abnormalities may allow for improved management of patients at risk of clonal evolution, and pharmacologic strategies to increase telomerase activity might mitigate the risk of cancer in these settings Disclosures: No relevant conflicts of interest to declare.

Donor-cell derived leukemia (DCL) is a rare but serious complication of hematopoietic stem cell transplantation (HSCT), reported in 5% of all relapses following allogeneic HSCT. Only 12 cases of DCL after umbilical cord blood (UCB) HSCT are reported in the literature. Multiple mechanisms have been proposed for DCL, including occult donor leukemia, impaired immunity, drug toxicity, or a bone marrow “leukemogenic milieu”. We propose a novel mechanism of leukemogenesis, mediated by very short telomeres of donor cells with subsequent severe telomere attrition in vivo, genomic instability, and progression to complex cytogenetics acute myeloid leukemia (AML), based on study of a patient who underwent UCB HSCT for myelodysplasia (MDS). The patient was a 44-year-old woman, subsequently shown to have a T354M mutation in GATA2, who presented at age 19 years with multiple infections and MDS. At age 41, due to progression of MDS to AML, she received induction chemotherapy and underwent a single 4/6 HLA-matched UCB HSCT. She had delayed engraftment. achieving an absolute neutrophil count of 500/ul more than 100 days post-transplant. Chimerism studies demonstrated 100% donor cell at all time-points post-transplant. Two years and eight months after HSCT she presented with pancytopenia, circulating myeloblasts, and 50% myeloblasts in bone marrow, indicating recurrence of with AML. However, cytogenetics revealed complex abnormalities, t(2p;3q) and an interstitial deletion of 5q, male cells, indicating donor origin of AML. Telomere length of the transplanted cord blood cells measured by Q-PCR showed a severe decrease in length to 7.7 kb compared with an average length of 13 kb in control UCB (n=12). Moreover the telomere length decreased precipitously to 6.6 kb 2 years after transplantation and 5.6 kb at the time of diagnosis of AML. Single telomere length assay (STELA) was used to assess chromosome-specific telomere length. Very short telomeres ( Telomere attrition occurs in the first year after HSCT, presumably due to proliferative stress on the stem cell compartment. In comparison to bone marrow, UCB have longer telomeres, but HSC number may be lower than in a BM inoculum, especially for adult recipients. In our case, initial telomere length of UCB was extremely short, for unclear reasons but possibly related to low cord blood cell viability. The long time that was needed for engraftment generated additional telomere attrition with accumulation of very short telomere and genomic instability leading to development of leukemia. Screening UCB for telomere length prior to HSCT might prove useful in identifying situations at risk for graft failure or malignant transformation. Disclosures: No relevant conflicts of interest to declare.

Abstract 1265 BACKGROUND: Acquired severe aplastic anemia (SAA) is a human disease characterized by severe pancytopenia due to marked reduction in the numbers of hematopoietic progenitors and stem cells. Immunosuppressive treatment with antithymoglobulin and cyclosporine results in improvement in blood counts in above 2/3 of patients. The most serious long-term complication in SAA is progression to myelodysplasia (MDS) and acute myeloid leukemia, usually associated with the cytogenetic abnormality monosomy 7. Clonal evolution usually requires a hematopoietic stem cell transplant and it is the major cause of morbidity and mortality. OBJECTIVE: Decreased average telomere length in leukocytes on presentation has been associated with increase risk of progression to MDS (Scheinberg et. al., 2010 JAMA, vol. 304(12):1358–64). Single telomere length assay (STELA) uses single molecule polymerase chain reaction (PCR) to amplify chromosome-specific telomeres based on the specificity of their subtelomeric region (Baird et. al., 2003 Nature Genetics, vol. 33(2):203–7). We have utilized STELA for identify SAA patients at risk of progression to MDS prior to development of the cytogenetic abnormality. METHODS: Peripheral blood samples were obtained at diagnosis and sequentially after immunosuppressive treatment from SAA patients who fulfilled entry criteria for protocols at the National Institutes of Health. Chromosome-specific PCR amplification of telomere using primers specific for Xp/Yp subtelomeric regions was performed. A non-radioactive, digoxin-labeled telomeric probe was used for Southern blotting to identify discrete bands, allowing quantification of very short telomeres. Results were confirmed using chromosome-specific primers for 2p, 11q, 12q, and 17p. RESULTS: Initially, multiple serial samples from five patients who had progressed to monosomy 7 MDS were analyzed by STELA of DNA extracted from peripheral blood leukocytes at the diagnosis of SAA and again at progression to MDS. All five patients had increase in very short telomeres ( CONCLUSIONS: This novel, non-radioactive method of quantifying chromosome-specific telomere length is capable of identifying very short telomeres in SAA patients progressing to MDS with monosomy 7. Previous studies have associated average short telomere length with chromosomal instability and progression to malignant phenotype. Identifying very short telomere in a subpopulation of circulating cells by STELA regardless of the average telomere length by qPCR is a more sensitive method. Chromosome-specific primers and analysis of end-to-end fusion of different chromosomes will allow for better characterization of the chromosomal instability leading to malignant transformation in these patients. Allogeneic stem cell transplant is the definitive treatment for MDS with monosomy 7 but requires time for identification of a suitable donor. Identification of critically short telomere before development of the cytogenetic abnormality will allow for timely management of patients at risk of clonal evolution. Therapeutic telomerase upregulation by androgens might reduce very short telomeres and potentially decrease the transformation risk. Disclosures: No relevant conflicts of interest to declare.

In the past, experimental results of the sound‐pressure level (SPL) were obtained which showed that the indoor sound field is regarded as a time‐variant system due to an air current. Using the time‐variant model represented by the direct sound and the varying delay time of single reflection, an experiment was performed on an effect on subjective preference. This experiment was focused on the effect of the modulation interval (Δ) of the delay time of the single reflection. The delay time of reflection (Δt1) was adjusted to the most preferred delay time of reflection which was obtained by the experiments for the time‐invariant sound field. In the case of fast tempo music, where the minimum value of the effective duration of the autocorrelation function is τemin=43 ms, the scale value of subjective preference is higher in a time‐variant sound field than a time‐invariant one. On the other hand, in the case of slow tempo music, τemin=127 ms, such an effect could not be found. It is suggested that a time‐varian...

The sound field in a concert hall is, rigorously speaking, a time‐variant system due to changes in temperature and air current. These time‐variant factors cause sound propagation that results in the fluctuation and sound‐pressure level. This paper shows that such a fluctuation may be identified by the autocorrelation function (ACF). As is widely known, effective duration of ACF (τe) of a sound source, which is defined by the delay at which the envelope of the normalized ACF becomes 0.1, describes several important subjective attributes [Y. Ando, Concert Hall Acoustics (Springer‐Verlag, Heidelberg, 1985)]. In this paper, values of τe of a music signal with changing tempo, dynamics, and articulation produced by a computer‐controlled piano were analyzed. Results indicate that the measured values of τe in an existing hall are shorter than those of the source signal itself, while τe values of reverberant sound field simulated as a time‐invariant system are 1.2–1.6 times longer than that of the source signal. T...

Background: Telomeres are repeated sequences (the hexanucleotide TTAGGG in vertebrates) located at chromosome ends of eukaryotes, protecting DNA from end joining or degradation. Telomeres become shorter with each cell cycle, but telomerase, a ribonucleoprotein complex, alleviates this attrition. The telomerase RNA component (TERC) is an essential element of telomerase, serving as a template for telomere elongation. The H/ACA domain of TERC is indispensable for telomere biogenesis. Mutations in the telomerase components allow accelerated telomere loss, resulting in various disease manifestations, including bone marrow failure. To date, this is the first detailed report of an H-box mutation in TERC that is related to human disease. Case presentation: A 26-year-old man with myelodysplastic syndrome (MDS) had very short telomeres. Sequencing identified a single heterozygous mutation in the H box of the patient's TERC gene. The same mutation was also present in his father and his son, demonstrating that it was germline in origin. The telomere length in the father's blood was shorter compared to age-matched healthy controls, while it was normal in the son and also in the sperm cells of the patient. In vitro experiments suggested that the mutation was responsible for the telomere shortening in the patient's leukocytes and contributed to the pathogenesis of bone marrow failure in our patient. Conclusion: We analyzed a mutation (A377G) in the H box of TERC in a young MDS patient who had significantly short-for-age telomeres. As telomeres protect chromosomes from instability, it is highly plausible that this genetic lesion was responsible for the patient's hematological manifestations, including marrow failure and aneuploidy in the hematopoietic stem cell compartment.

Digital television is transforming both broadcasting and, as a result of convergence, the larger world of communications. The impending analogue switch-off will have a major impact on households all over the developed world. Digital Broadcasting considers the effects of digital television on the availability, price and nature of broadcast services in the Americas, Europe and Japan. It shows how this depends upon what platforms – cable, satellite, fixed or wireless broadband – countries have available for use and also upon government policies and regulatory interventions.