•We generated CDA patient-derived iPSCs carrying the KLF1 E325K mutation (CDA-iPSCs).•We developed an inducible expression system of KLF1 E325K using CDA-iPSCs.•We found that KLF1 E325K induced G1 ...cell cycle arrest at the CD71+/CD235a+ stage.
Krüppel-like factor 1 (KLF1), a transcription factor controlling definitive erythropoiesis, is involved in sequential control of terminal cell division and enucleation via fine regulation of key cell cycle regulator gene expression in erythroid lineage cells. Type IV congenital dyserythropoietic anemia (CDA) is caused by a monoallelic mutation at the second zinc finger of KLF1 (c.973G>A, p.E325K). We recently diagnosed a female patient with type IV CDA with the identical missense mutation. To understand the mechanism underlying the dyserythropoiesis caused by the mutation, we generated induced pluripotent stem cells (iPSCs) from the CDA patient (CDA-iPSCs). The erythroid cells that differentiated from CDA-iPSCs (CDA-erythroid cells) displayed multinucleated morphology, absence of CD44, and dysregulation of the KLF1 target gene expression. In addition, uptake of bromodeoxyuridine by CDA-erythroid cells was significantly decreased at the CD235a+/CD71+ stage, and microarray analysis revealed that cell cycle regulator genes were dysregulated, with increased expression of negative regulators such as CDKN2C and CDKN2A. Furthermore, inducible expression of the KLF1 E325K, but not the wild-type KLF1, caused a cell cycle arrest at the G1 phase in CDA-erythroid cells. Microarray analysis of CDA-erythroid cells and real-time polymerase chain reaction analysis of the KLF1 E325K inducible expression system also revealed altered expression of several KLF1 target genes including erythrocyte membrane protein band 4.1 (EPB41), EPB42, glutathione disulfide reductase (GSR), glucose phosphate isomerase (GPI), and ATPase phospholipid transporting 8A1 (ATP8A1). Our data indicate that the E325K mutation in KLF1 is associated with disruption of transcriptional control of cell cycle regulators in association with erythroid membrane or enzyme abnormalities, leading to dyserythropoiesis.
Introduction: Blood donor recruitment and retention in the younger generation is an important concern in several countries with an aging population. In Japan, which is going to have an aging ...population, the number of blood donors has decreased by 15% over the past decade. As such, drastic measures must be taken to maintain the necessary blood stocks. The promotion of blood donation in high schools has been attempted in many countries. This strategy is particularly attractive because successful recruitment of young donors will ensure long term supplies of blood are maintained. To enhance the effectiveness of this approach it is important to communicate the need for blood donation by high school students and conduct appropriate surveys. Although there have been several reports on the promotion of blood donation by young people, these have not generally analyzed their psychology, personal environment and the views of large numbers of high school students.
Materials and Methods: The study was accepted with each high school staff meeting and IRB in our university (#25-159). Inquiry anonymous surveys were designed for high school students, who answered by their own volition. The survey included 50 questions as follows; gender; age; build; previous blood donation by the individual as well as family members and friends; lifestyle; diet; views concerning blood demand in society; location of blood donation centers; knowledge of blood donation methods; blood recovery after donation; reasons for declining to give blood; ideas for an effective campaign to recruit blood donors; previous education on blood donation in their school etc. Individual views concerning an effective campaign to recruit new donors, events, characters and media were given in a free style. This work was supported by the Japanese Ministry of Health, Labor and Welfare (H25-medicine-general-022).
Results:We obtained answers from 94% of students surveyed (16,333). The first and second studies were done in 2013 and 2014, and included 8,456 and 7,877 students, respectively. In the first study, the male/female ratio was 0.88. 1% and 26% of student body weights were <40kg and <50kg, respectively. 8% of students had at least one experience of blood donation. 3% of the students could not donate blood due to medical reasons (e.g., anemia). 62% of the students often experienced feelings of fatigue in their daily life. 5% of the students suffered sleeplessness. 31% of the students believed in the existence of synthetic blood. 54% of the students did not know the location for blood donation. Only 18% of the students understood the process of blood donation in outline. Moreover, only 29% of students knew the age limit for blood donors. 32% of the students were aware of the decrease in blood donation and the future inadequacy of blood supplies. 66% of the students had previous volunteer experience. 48% of the students misunderstood the risk of contracting a viral infection by blood donation. 26% of the students feared pain caused by needles. 51% of students requested more information from schools and other institutions. Suggested ways to promote blood donation included television (21%), school classrooms (19%), Twitter or Facebook (11%), mobile blood donation units (11%), software applications on cellular phones (8%) and newspaper advertisements (6%). A considerable number of respondents suggested an effective way to recruit more volunteers was to carry out this kind of survey amongst students.
Conclusions: Our survey provided a lot of information concerning blood donation from high school students, which will be applicable to a number of countries. The study clarified that many students lack sufficient information about blood donation and transfusion. Moreover, a considerable number of students suffer a lot of stress and fatigue in their daily life. Substantial time and effort must be devoted to educating the student population on the need for blood transfusion, as well as the safety and risk factors associated with blood donation. Moreover, their psychology, daily life and views concerning transfusion medicine must be better understood.
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No relevant conflicts of interest to declare.
The association of ethnicity with the incidence of graft-versus-host disease (GVHD) and other clinical outcomes after transplantation is controversial. We compared the results of HLA-identical ...sibling bone marrow transplantations for leukemia, performed between 1990 and 1999, among different ethnic populations, including 562 Japanese, 829 white Americans, 71 African Americans, 195 Scandinavians, and 95 Irish. Results for adults and children were analyzed separately. Multivariate analyses of adult patients showed that white Americans, African Americans, and Irish cohorts were at significantly higher risk for acute GVHD than Japanese or Scandinavian cohorts (relative risk RR = 1.77, P < .001; RR = 1.84, P < .006; RR = 2.22, P < .001, respectively). White Americans, African Americans, and Irish, but not Scandinavians, were at significantly higher risk for early (within 3 months of transplantation) transplant-related mortality (TRM) compared with Japanese (RR = 2.99, P < .001; RR = 5.88, P < .001; RR = 2.66, P < .009, respectively). No differences in the risk for chronic GVHD, relapse, and overall survival were noted. In the pediatric cohort (limited to Japanese and white Americans), white Americans were at significantly higher risk for acute (RR = 1.93; P = .04) and chronic (RR = 3.16; P = .002) GVHD. No differences in other clinical outcomes were noted. Our findings suggest that ethnicity may influence the risk for GVHD, though overall survival rates after transplantation remain similar.
Background Although intravenous immunoglobulin (IVIG) is widely used in clinical practice, there are no official guidelines for its proper usage in Japan. We have started the preparation of ...guidelines for appropriate usage of IVIG, firstly by collecting clinical cases treated with IVIG nationwide. Methods Questionnaires concerning the number of patients treated with IVIG and the amount of IVIG used by disease and age (children and adults) from July to September 2003 were sent to 732 hospitals with more than 400 beds. Results Replies were obtained from 319 hospitals, of which 268 (36.6%) were suitable for data analysis. The number of patients and the amount of IVIG used were 1, 955 cases and 28, 684g for children and 6, 615 cases and 142, 334g for adults, respectively. Main diseases in children and adults were infectious diseases (including post-operative infection) in 33% and 73%, autoimmune diseases in 28% and 10%, and immunodeficiency diseases in 16% and 8%, respectively. The differences between them were significant. In particular, Kawasaki disease in children was predominant among autoimmune diseases, representing 44.5% of cases. The amount of IVIG used in children was 60% for autoimmune diseases and 17% for immunodeficiency diseases, while that in adults was 45% for infectious diseases and 35% for autoimmune diseases. Pathogens in these infectious diseases of both children and adults were bacteria in more than half of the cases, and high CRP values as a factor of severe infection were 28% in children (more than 10mg/dl) and 51% in adults (more than 15mg/dl). Conclusions These results indicate that IVIG is mainly used in both autoimmune diseases and infectious diseases, and that IVIG is also used in some non-severe infectious diseases.
The Japan Society of Blood Transfusion (JSBT) organized a Board for certification of medical doctors in Transfusion Medicine (Board certified medical doctors by the JSBT) and a Board for ...certification of medical technologists for transfusion medicine (qualified medical technologists in Transfusion Medicine). For certified medical doctors the JSBT co-ordinated with the Japanese Association of Medical Science and with the Japan Medical Association, and for qualified medical technologists the JSBT co-organized with the Japanese Society of Laboratory Medicine, the Japanese Association of Medical Technologist and the College of Clinical Pathology of Japan. By May 2001, 259 of the certified medical doctors and 875 of the qualified medical technologists have been certified and are participating actively to increase the level of transfusion practice at individual facilities.
In hematological diseases such as myeloproliferative disorders (MPD) or myelodysplastic syndromes (MDS), some abnormalities in the chemiluminescence of neutrophils are observed. There are two groups; ...one includes chronic myelogenous leukemia (CML), essential thrombocythemia (ET) and MDS, which all have decreased chemiluminescence of neutrophils. The other group includes polycythemia vera (PV) which has increased neutrophil chemiluminescence. We studied the neutrophil function by analyzing the chemiluminescence in 35 patients with hematological diseases. In most of these cases the defects in chemiluminescence in response to N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) were correlated with those in response to phorbol 12-myristate 13-acetate (PMA). But there were exceptional cases in which the maximal light emission of chemiluminescence (Max CL) in response to FMLP was obviously lower than controls despite the fact that the Max CL in response to PMA was the same as the controls. These facts suggest a heterogenicity of the defect site in these diseases. There was a correlation between the level of chemiluminescence and the neutrophil alkaline phosphatase (NAP) activity in these patients. In vitro culture of CML neutrophils with granulocyte colonystimulating factor (G-CSF) showed a correlation between the increase in the level of chemiluminescence and NAP activity. These results suggest that NAP may take part in the control of neutrophil function. (Internal Medicine 32: 204-209, 1993)
: In recent years the procedures for peripheral blood mononuclear cell (PBMNC) harvests have gradually been increasing. These PBMNCs are collected for several treatments, for example, donor ...lymphocyte infusion (DLI), immunotherapy for solid carcinoma, and regeneration therapy for ischemic limbs. In order to analyze the optimal procedure for collecting PBMNCs safely and efficiently, we evaluated 129 PBMNC apheresis procedures from April 1996 to May 2003, without hemopoietic stem cell mobilization by G‐CSF. In every case, PBMNC collections were performed with a COBE Spectra cell separator (Gambro BCT). The median apheresis volume was 5550 mL. The median of blood draw speed was 48.1 mL/min. The median TNC (total nuclear cell) number in products was 50.4 × 103/µL. In the regression analysis, no significant correlation was seen between the blood draw speed and the concentrations of TNC in products (Y = aX + b, a = 0.842497, b = 17.11352, r = 0.222032, P = 0.012464). A positive correlation was seen between WBC on apheresis day and the concentrations of TNC (Y = aX + b, a = 0.009822, b = 3.224679, r = 0.550431, P = 2.93 × 10−11). A significantly higher correlation was seen between the MNC (mononuclear cells) on apheresis day and the concentrations of TNC (Y = aX + b, a = 0.028278, b = 13.09266, r = 0.696988, P = 9.486 × 10−9). This study has shown evidence that a higher increment of blood draw speed does not provide a higher concentration of products. An adequate apheresis speed is about 40 mL/min. If we want to obtain sufficient cell counts, it is very important to obtain sufficient volume with a moderate blood draw speed, therefore protecting against side‐effects.
A system for educating inspectors and accrediting hospitals for transfusion medicine (I&A System) did not become widespread under its former regulations. To clarify its significance in Japan, we ...examined the approximately 3-year outcomes after a relaunch of the system by the Japan Society of Transfusion Medicine and Cell Therapy.The study involved 100 hospitals: 49 newly accredited and 51 re-accredited during the period from January, 2016 to March, 2019. The distribution of each of the accredited facilities and items for which improvement was advised upon inspection was examined. 2019.A large number of accredited hospitals belonged to the Tohoku or Tokai Branch of the society. The mean number of mandatory and important items for accreditation for which improvement was advised upon inspection of the newly accredited hospitals was 2.6 and 5.3, respectively. Among the mandatory items requiring improvement, those related to the storage and management of blood products were the most frequent, followed by double-checking the blood type, and management of adverse events when collecting autologous.blood. Among the important items requiring improvement, those related to the management of and countermeasures against adverse events were the most frequent. The mean number of items for which there was a discrepancy between the self-inspection results and inspectors' opinion was 6.0. This suggests the importance of third-party inspections.Our findings suggest that the I&A System was useful for improving medical safety, which is important for transfusion medicine. In the future, we will more actively disseminate useful information from an inspector/person concerned with accrediting facilities to healthcare professionals while promoting the sharing of this information with those engaged in transfusion medicine. We will also develop I&A programs that will further enhance the safety of blood transfusion.
Five patients (six courses) with refractory idiopathic thrombocytopenic purpura (ITP) were given 6 million units of recombinant interferon (IFN) alpha-2b in 12 doses to achieve an improved response ...rate compared to previous studies using 3 million units. From the initial IFN administration, the platelet count increased from a pre-treatment level of 20.7±17.7×103/μl (mean±SD) and reached its first peak in weeks 2 or 3 of therapy (p<0.05). In week 5, the platelet count made its second and maximum peak (66.5±57.9×103/μl ; p<0.05). A relatively good response of the platelet count (an increase to >50×103/μl) was observed in three patients (four courses) out of five. These responses were not much faster or more improved than in previous reports, and a dose of 6 million units may be too large to treat some ITP patients. The platelet-associated IgG level showed a tendency to be reduced with IFN therapy. The mechanism for the increase of the platelet count may be the modification of platelet autoantibody production. (Internal Medicine 31 : 1343-1347, 1992)
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