bluuebunny/crossref_metadata_2025_split huggingface dataset:
High Probability of Survival after Related and Alternate Donor Hematopoietic Cell Transplantation for Pediatric Patients with High Risk AML Using a Fludarabine Based Preparative Regimen
Authors: Tan Poh-Lin , Yeoh Allen , Quah Thuan-Chong , Kurtzberg Joanne , Wagner John E. 
Published: April 2020

Abstract
Hematopoietic cell transplantation (HCT) in pediatric patients with high risk acute myeloid leukemia (AML) such as those with non-Down de novo acute megakaryoblastic leukemia (AMKL), transformed myelodysplastic syndrome (MDS), therapy-related AML (tAML) and refractory/resistant AML (rAML) has been limited by graft failure, regimen related toxicity (RRT) and relapse. Between September 2005 and July 2008, 10 pediatric patients, median age of 7.6 (range, 1.3 – 17.0) years with such high risk AML (AMKL, n=2; MDS/AML, n=4; tAML, n=1; rAML; n=3) received a novel Fludarabinebased preparative regimen followed by T cell replete transplantation with bone marrow (BM, n=4), peripheral blood stem cell (PBSC, n=5) or umbilical cord blood (UCB, n=1) in a Singapore Pediatric BMT center. Seven patients received stem cells from alternate donors (AD) and 3 from matched related donors (MRD). Prior to HCT, 6 patients were in 1st complete remission (CR1), 2 in CR2 and 2 had progressive diseases. AD consisted of single antigen mismatched related and non-genoidentical unrelated donors (antigenic match at HLA-A, B, DRB1*). All patients received IV FLU 125 mg/m2, BU, 0.8 – 1.2 mg/kg 6 hourly, MEL, 140mg/m2. AD recipients received additional ATG (Thymoglobuline) 5mg/kg. GVHD prophylaxis consisted of cyclosporine and methotrexate or mycophenolate motfetil depending on the stem cell source. Excluding the 2 patients who entered HCT with progressive diseases, all other patients achieved myeloid engraftment at a median of 14 (range, 9 – 16) days and achieved sustained stable full donor chimera from 21 days after HCT. The median duration and costs of HCT admissions was 34 (range, 17 – 66) days and USD 51,317 (range, 41,416 – 155,905), respectively. Common post-HCT complications included engraftment syndrome (n=4) and reactivation of Herpes viruses (n=4). There was no severe RRT. Incidences of grade I–III acute GVHD and chronic GVHD were 63% and 67%, respectively. All these patients are alive in sustained CR at a median follow-up of 303 (range, 44 – 1037) days with Karnosky/Lansky scores of 90 – 100%. Both the patients with progressive diseases had graft failures and subsequently died of disease at a median of 138 (range, 77 – 198) days after HCT despite experimental salvage therapy with allogeneic natural killer cell transplantation from parental donors. In summary, this novel FLU based preparative regimen paired with non-genoidentical HCT is safe, inexpensive, allows prompt engraftment, and appears to provide anti-leukemia effects in pediatric patients with high risk AML entering HCT in remission. Long term follow-up will determine relapse risks and whether this non-irradiation based regimen is associated with a reduction in late effects including malignancy.
Laser Transmyocardial Revascularization (TMR) Combined with Autologous Bone Marrow Cells for Diffuse Coronary Disease: Immunophenotype Analysis and Preliminary Clinical Results in 14 Cases
Authors: Alegre A. , Aguado B. , Alfonso-Pérez M. , Muñoz C. , de Landázuri M. Ortiz , Sánchez-Madrid F. , Cámara C. , Lara F. , Duarte J. , Reyes G. 
Published: April 2020

Abstract
Introduction: Stem cell (SC) based therapy in ischemic heart disease is actually in development with the aim of improving cardiac regeneration. The way of administration, source of SC and the mechanisms of its clinical effects are yet not well defined. We present preliminary results of a group of patients treated with laser revascularization and autologous bone marrow (BM) cells administration, evaluating cells immunophenotype and clinical results.
Patients and methods: Fourteen patients (57–78 y) with diffuse coronary disease and medically refractory class III/IV angina were prospectively evaluated for cell-based therapy combined with laser transmyocardial revascularization (TMR) PHOENIX-TM (Cardiogenesis, Irvine, CA, USA). BM mononuclear cells (MC), were obtained by BM aspiration (120 mL) and processed for concentration using HARVEST-TM system (Harvest Technologies Inc. GMBH, Munich). Immunophenotypic characterization of the final infused product was performed by standard four-color flow cytometry with monoclonal antibodies directed against the human surface antigens CD29, CD49a, CD105, CD106, CD117, CD34, CD133 and CD45, to evaluate different cell populations.
Results and conclusions: Patients received a median of 56.8×106 (43.7–79) BMMC/ mL 0.5×106 (0.05–1) CD34+ cells/mL and 0.23×106 (0.001–0.7) CD133+ cells/mL. All the patients are alive after this treatment: median of 9 months, (1–15), with improvement of their cardiological clinical status and QoL. As observed (Table 1) the BMMC used in the therapy contained a significant proportion of CD133+ and CD34+ hematopoietic SC. In addition, the samples contained cells with CD105+ CD45− or CD29+ CD45− CD34− phenotypes which have been described as mesenchymal stem cells (MSC). Bone marrow CD106+ positive cells represent a multipotent subpopulation of MSC including endotelial. Correlation between the relative quantity of the different cell subsets and clinical results of the patients will be presented.
TABLE 1
CD34 CD133 CD117 CD29 CD49a CD105 CD106 Mean 3.53 2.42 7.37 2.23 0.75 6.75 0.33 Std. Deviation 1.72 1.68 2.60 0.97 0.75 3.34 0.23 25% Percentile 1.95 0.85 4.95 1.25 0.15 3.10 0.2 Median 3.20 2.15 6.70 2.30 0.40 7.05 0.25 75% Percentile 5.45 4.25 10.45 3.15 1.70 10.10 0.55
Mini-Autologous Hematopoietic Stem Cell Transplantations as a New Treatment Modality for Multiple Sclerosis Patients
Authors: Novik Andrei A , Melnichenko Vladimir Y , Fedorenko Denis A , Kruglina Ruslana V , Ionova Tatiana I , Kishtovich Anton V , Ivanov Roman I , Gorodokin Gary J 
Published: April 2020

Abstract
Conventional therapies do not provide satisfactory control of multiple sclerosis (MS) due to their inability to eradicate self-specific T cell clones. Recently, high-dose immunosuppressive therapy with autologous hematopoietic stem cell transplantation (AHSCT) was proposed as a new and promising therapy for MS patients. Taking into account the information about high risk of transplant-related mortality and severe side effects of myeloablative conditioning regimens, the rationale to use non-myeloablative regimens sounds reasonable. The goal of our research was to study the safety and efficacy of mini-AHSCT in MS patients. Fifty three patients with MS (secondary progressive – 16 patients, primary progressive – 12, progressive-relapsing – 3 and relapsing-remitting – 22) were included in this study (mean age - 30.0, range: 17–47; male/female – 23/30). The conditioning regimen included reduced or modified BEAM. Median EDSS at base-line was 4.0 (range 1.5 – 8.0). The median follow-up duration was 7 months (range 6 – 20 months). Neurological and quality of Life (QoL) evaluation was performed at baseline, at discharge, at 3, 6, 9, 12 months, and every 6 months thereafter AHSCT. MRI examinations were performed at baseline, at 6, 12 months, and at the end of follow-up. Transplantation procedure was well tolerated by the patients with no transplant-related deaths. Among 29 patients included in the efficacy analysis 15 patients (52%) experienced clinical stabilization; 14 (48%) – improvement. EDSS improved by 0.5 points in 11 patients, by 1.0 points in 2 patients, by 2.0 points in 1 patient. One patient progressed after 6 months stabilization. No active, new or enlarging lesions were registered in patients without disease progression. Out of 27 patients included in QoL analysis 21 exhibited QoL improvement and 4 – stabilization during the follow-up; QoL worsened in 2 patients. All the patients without disease progression were off therapy throughout the post-transplant period. In conclusion, mini-AHSCT may be considered as a new, safe and effective treatment for MS. The results of mini-AHSCT in MS patients are promising both in terms of clinical and patient-reported outcomes. The collection of long-term follow-up data is worthwhile to verify these findings. The rationale of evolution from myeloablative to non-myeloablative transplant regimens should be confirmed by the further studies.
Genome-Wide Scanning of Epigenomic Alterations in Childhood Leukemia
Authors: Zhong Sheng L. , Xiao Yuanyuan , Yeh Ru-Fang , Zhou Mi , Wrensch Margaret R , Zheng Shichun , Wiencke John , Wiemels Joseph L. 
Published: April 2020

Abstract
To study how epigenetic changes contribute to childhood leukemia, and to identify new disease biomarkers, we combined an approach of pharmacologic inhibition of epigenetic modifications of chromatin with gene expression arrays (Affymetrix Exon 1.0; and miRNA array, UCSF-designed) to reveal precise epigenetic aberrations on a global scale in defined subsets of leukemia cell lines that recapitulate common genetic subtypes of leukemia. From these analyses, we identified &gt;200 genes up-regulated by 5-aza-2′-deoxycytidine and trichostatin A treatment (including genes encoding both mRNA and miRNA). We validated 50 selected genes using real-time quantitative PCR. We also tested promoter regions for methylation in the cell lines and selected pediatric leukemia bone marrows using bisulfite-DNA sequencing. A panel of leukemia-specific methylation targets was discovered, which includes some well-known genes (such as HOXA5 and DAPK) and several novel candidates in childhood leukemia (eg., CD82 and KLF4). To extend these observations, we performed methylation microarrays on DNA samples isolated from the studied cell lines as well as bone marrows of 35 childhood leukemia patients with defined cytogenetics, using Illumina Methylation Cancer Panel I. This panel includes a total of 1,505 CpG loci from 807 genes. We also assayed corresponding archived newborn bloods (Guthrie cards) from 32 cases, and 19 control Guthrie cards of children matched to the cases based on gender and age, to guage the impact of congenital methylation events on childhood leukemia. Many targets genes, including known tumor suppressor genes and genes involved in DNA repair, cell cycle control and differentiation were found to be methylated in leukemia cell lines and the leukemic bone marrow samples, but not in Guthrie card controls. In addition, the methylation status of a core group of 67 CpG sites can differentiate the common subtypes of childhood leukemia – those with TEL-AML1, E2A-PBX1, high hyperdiploidy, and normal karyotypes. Several genes concordantly methylated in Guthrie cards and leukemic bone marrow may represent prenatal epimutations. With the integration of such assays, we have constructed a panel of potential candidate genes and miRNAs involved in development of childhood leukemia which have aberrant promoter methylation. Our studies lay a foundation for a comprehensive understanding of the full extent of epigenetic changes in development of childhood leukemia and how they may be exploited for therapeutic purposes as well as for early detection of this disease.
Differente Patterns of Telomerase Reverse Transcriptase (hTERT) mRNA Variant Expression Profiles in Patients with Myelodysplastic Syndromes (MDS) and Acute Myelogenous Leukemia (AML)
Authors: Verrou Evgenia , Kaloutsi Vassiliki , Katodritou Eirini , Garypidou Vasilia , Nikitidou Anna , Gastari Vassiliki , Lazaridou Anna , Zervas Konstantinos , Kotoula Vassiliki 
Published: April 2020

Abstract
BACKGROUND: Alternative splicing of hTERT mRNA contributes to the regulation of telomerase activity in normal and neoplastic cells. The deleted variant A (Adel), lacking 36nts in the beginning of exon 6, blocks telomerase activity while the B deleted variant (Bdel), lacking exons 7 and 8, exhibits no action in terms of telomerase activation. The expression of hTERT mRNA has been investigated in myeloid malignancies with controversial results. The expression of hTERT mRNA variants has not been explored in MDS patients.
AIM: To investigate the expression of hTERT mRNA variant transcripts A+B+ (contained in the full-length product), Adel and Bdel in the bone marrow of MDS and AML untreated patients.
METHODS: Bone marrow aspirates from 27 patients with refractory anaemia or refractory cytopenia with multilineage dysplasia (RA or RCMD, defined as group A), 13 patients with refractory anaemia with excess of blasts (RAEB1 or RAEB2, defined as group B) and 17 AML patients (defined as group C) were studied. Relative expression of the above transcripts was assessed on total RNA with reverse transcription followed by real-time polymerase chain reaction (PCR). The data obtained were analyzed by using Pearson chi-square and Kruskal Wallis statistics. All individuals signed informed consent before sampling.
RESULTS: hTERT mRNA was detected in 10, 4 and 3 patients of groups A, B and C respectively. The full length product (A+B+) was detected in groups A, B and C (2/27, 4/13, 3/17 respectively) (pA/B=0.053). Adel isoform was detected in groups A (7/27) and B (3/13) but not in group C (0/17) (pA/C=0.022). Bdel isoform was observed in group A (3/27), B (3/13) and C (2/17). Co-expression of isoforms was observed in 2 patients of A group (Adel/Bdel, Adel/A+B+), 4 patients of group B (1 Adel/A+B+, 1 Bdel/A+B+, 2 Adel/Bdel/A+B+) and two patients of group C (Bdel/A+B+). Relative expression levels of Adel, Bdel, and A+B+ transcripts were not significantly different among patient groups.
CONCLUSIONS: Alternatively spliced hTERT variants either lack a critical reverse-transcriptase motif or produce a non-functional reverse-transcriptase. Therefore, cells may control telomerase activity by switching their hTERT mRNA variant expression profile. Our results indicate that aberrations in hTERT variant profiles, especially regarding the Adel isoform, may be implicated in the pathogenesis and progression of MDS. The increased incidence of Adel positivity among RA or RCMD patients may be connected with a decline in telomerase activity and therefore contribute to the establishment of chromosomal abnormalities. This observation underlines the need to distinguish between the different hTERT mRNA variants when investigating hTERT expression in MDS patients.
Upregulation of hGC-1 Expression by Hypomethylation, Retinoic Acid and Interferons in Human Myeloid Leukemia
Authors: Liu Wenli , Rodgers Griffin P 
Published: April 2020

Abstract
hGC-1 (Granulocyte colony stimulating factor induced gene-1, also called GW112 and olfactomedin 4) was first identified in human myeloid precursor cells induced by granulocyte colony stimulating factor (G-CSF). It is a myeloid lineage and differentiation stage specific gene. Its expression, regulation and biological function, specifically in myeloid cells, are still poorly understood. In this study, we analyzed the hGC-1 gene expression in leukemia patients and further investigated the mechanism of hGC-1 gene regulation in leukemia cells. We found that hGC-1 was overexpressed in myeloid leukemia patients compared with normal individuals in peripheral blood leukocytes (p&lt;0.01) and its expression in accelerated phase of chronic myeloid leukemia (CML) patients was significantly higher than that in chronic phase (p&lt;0.01) using a dot blot and quantitative RT-PCR analysis. Hypomethylation of CpG sites in the promoter of hGC-1 gene were observed in CML patients by pyrosequence and 5-aza-2′-deoxycytidine induced hGC-1 expression in myeloid leukemia cells, suggesting that promoter CpG methylation status affects the expression of hGC-1 gene. All-trans-retinoic acid (ATRA) and interferons (IFNs) are active anti-leukemia agents. ATRA and IFNs have shown synergistic interactions in various experimental conditions and represent a potentially useful therapeutic combination in the treatment of various types of leukemia. However, the target genes and molecular basis of these interactions still needs to be further elucidated. Here, we identified that hGC-1 was a target gene of RA in myeloblastic leukemia cells. Treatment with ATRA induced hGC-1 expression in HL-60 cells and enhanced hGC-1 expression in AML-193 and GDM-1 cells. Deletion analysis led to the identification of a positive retinoic acid response element (DR5) and a negative response element (DR1) within hGC-1 promoter. Furthermore, electrophoretic mobility-shift assays demonstrated that RARa/RXRa binds to the DR5 site. Transfection study in COS-7 cells revealed RARa/RXRa mediated the RA induced transactivation of hGC-1 promoter. We also found that hGC-1 was an early responsive gene of IFN a and b in myeloid leukemia cells (HL-60, AML-193 and GDM-1). An effective interferon-stimulated response element (ISRE) was identified in the promoter of hGC-1 gene by examining the deletion mutants in luciferase reporter gene assay. Combined application of ATRA and IFNa and IFNb enhanced hGC-1 expression synergistically. Taken together, hGC-1 is identified as a novel target gene of methylation modification, RA and IFNs in myeloid leukemia cells. Our results suggest that hGC-1 is a potential marker for CML stage and may play a role in retinoic acid and interferon induced biological effects in leukemia cells.
Clinical Significance of P16 in Langerhans Cell Histiocytosis
Authors: Kim Sun Young , Kim Hee Jin , Yoon Hoi Soo , Bae Keun Wook , Park Jeong A. , Im Ho Joon , Seo Jong Jin , Lee Mee Jeong 
Published: April 2020

Abstract
Background. The p16 protein has been identified as a tumor suppressor that functions by inhibiting the cyclin-dependent kinases CDK4 and CDK6. Deletions or point mutations in the p16 gene have been found in a number of human cancers, emphasizing its importance in regulating cell cycle progression. The authors have chosen to analyze P16 expression in biopsies from children with LCH and correlated it with clinical manifestation and outcome in a broad range of organs.
Patients and Methods: Archival paraffin block were retrieved from children diagnosed with LCH and followed up at Asan medical center and Chungnam National University Hospital between 1998 and 2007. Pathological Langerhans cells were identified morphologically associated with the characteristic inflammatory type infiltrate and were confirmed by immunohistochemical staining with antibodies to CD1a, and S100. The disease was classified as single system defined as uni- or multifocal involvement of a single organ system or multi-system defined as involvement of multiple organ systems with or without organ dysfunction. The slides were stained with P16 antibody and semi-quantitatively evaluated using 5 grade systems, negative means absence of staining, ±, weakly positive, +, similar with lymphocytes around the LCs, 2+, stronger than lymphocystes, 3+, much stronger than lymphocytes.
Results: The specimens from 57 patients (31 boys and 26 girls) were available for immunohistochemistry. Of these, 12 slides were not suitable because the quality of staining was not good for evaluation or the tumor cells are too difficult to be differentiated. The median age of 45 patients (24 boys and 21 girls) was 9.5 years, range 5 months to 22 years with a definite diagnosis of LCH based on CD1a positivity. The P16 protein was expressed in various degrees except one specimen. No significant difference was found between the level of P16 expression in lesions from patients with single-system, and multi-system disease. But higher tendency of relapse have shown in 1+, 2+ and 3+ groups compared with negative and weakly positive groups.
Conclusions: This study has shown a diffuse expression of P16 protein in both single and multi-system lesions of LCH. P16 might be involved in the pathogenesis of LCH.
GATA-1 Protein Levels Are Not Altered in CD34+ and CD61+ Cells, but NF-κb Protein Levels Are Elevated in Patients with Idiopathic Myelofibrosis
Authors: Wang Jen-Chin , Naik Seema , Chen Chi , Xiao Ying-Yi , Chang Tsong H. 
Published: April 2020

Abstract
GATA-1 and NF-kB are important in megakaryocytopoiesis and pathogenesis of idiopathic myelofibrosis (IMF). Previous studies have shown that GATA-1 levels were normal in mRNA but were lower in protein content in CD61+ cells of patients with IMF, and we reported previously that GATA-1 protein levels were not altered in CD61+ cells. The current study includes more patients and also measured NF-kB levels. ELISA assay (GATA-1 and NF-kB ELISA kit, Panomics Inc. CA) was used to determine and compare the protein content of nuclear extracts of blood hematopoietic progenitor cells (CD34+) and megakaryocytes (CD61+ cells) from patients with IMF, patients with other myeloproliferative disorders, and with controls. CD61+ cells were obtained from 10-day cultures of blood CD34+ cells in serum-free culture medium with thrombopoietin and stem cell growth factor. Our study confirmed that GATA-1 protein levels are not altered in megakaryocytes (cultured CD 61+ cells) in patients with IMF (Fig. 1) and NF-kB levels are increased (Table 1). We conclude that GATA-1 protein levels are not altered in patients with IMF compared with controls.
Fig 1. GATA-1 Protein levels in CD61+ cells Cultured from Patients with IMF & Others Fig 1. GATA-1 Protein levels in CD61+ cells Cultured from Patients with IMF & Others
Table 1. NF-kB Levels (ng/2μg protein)
IMF Controls CD34+ cells 1.39 ± 0.27 1.10 ± 0.24 CD61+ cells 1.54 ± 0.52 0.45 ± 0.22
Longterm Prophylactic Treatment with Recombinant Factor VIIa (NovoSeven) in Severe Congenital Factor VII Deficient Patients
Authors: Gerhardt Andrea , Scharf Rudiger E. , Zotz Rainer B. 
Published: April 2020

Abstract
Background: Treatment of congenital FVII deficiency consists of replacement therapy with plasmic FVII concentrates or recombinant FVIIa (rFVIIa). Relatively small amounts of rFVIIa are required for replacement therapy in FVII deficient patients. Because of its short half-life of approximately 3 hours rFVIIa has not been regarded as a routine prophylactic treatment option for FVII deficiency.
Case Reports: We report on our experience of prophylactic treatment with rFVIIa in two FVII deficient patients. Patient one is 43-year-old male with severe FVII deficiency (&lt;1% activity) associated with compound heterozygous Gly78Asp and heterozygous Cys194Tyr mutations. After birth, he experienced severe tissue bleeding complications and later on various joint bleedings with consecutive hemophilic arthropathy. He is now on treatment with rFVII (1.2 mg 3 times per week) since 28 months. Patient two is a 36-year-old male with FVII deficiency of &lt;1% FVII activity. He experienced recurrent mucosal bleeding episodes and large skin hematomas. His genetic defect consists in a homozygous missense mutation (1061 C&gt;T, Ala354Val), a heterozygous missense mutation (218 T&gt;A, Leu73Gln), and a heterozygous frameshift mutation (1391delC, Pro464HisfsX32). The patient is on prophylactic treatment with rFVII (1.2 mg 3 times per week) since 22 months. Since substitution with rFVII, no spontaneous bleeding episodes occurred and no side effects were observed in both patients. In conclusion, prophylactic treatment with rFVIIa in FVII deficient patients appears to be an effective and safe therapeutic option. The mechanisms by which rFVIIa prevents bleeding episodes despite a short half-life of 3 hours remains to be elucidated.