bluuebunny/crossref_metadata_2025_split huggingface dataset:
Outcome Assessment in Hemophilia: A Pilot Study on Correlation of Different Instruments.
Authors: Fischer Kathelijn , Van Genderen Frank R. , De Kleijn Piet , Van den Berg Marijke 
Published: April 2020

Abstract
Hemophilia is a rare disease, requiring life-long treatment with expensive clotting factor concentrates. Outcome assessment in hemophilia is important to establish both results of treatment, and compare between strategies. Since the 1980s, hemophilia specific assessment tools were available only for radiological (Pettersson score) and clinical (orthopedic joint score) joint damage. Recently, the HAL (Hemophilia Activities List) was developed to assess functional limitations, and the orthopedic joint score was adapted into the HJHS (hemophilia joint health score) to assess more subtle joint damage.
To assess correlation of instruments for outcome assessment, a pilot study was conducted in 22 young adults with hemophilia (15 severe, 5 moderate, 2 mild). Joint damage (Pettersson score, HJHS), physical function (self-reported: HAL, performance-based: Figure 8 test; time taken to walk around two pilons at a distance of 5 meter at maximum speed in a pattern of figure 8), Health Related Quality of Life (SF36), and utility (EQ5D) were measured.
Median age was 17.8 years (range 14–30), 15 (68%) patients were treated with early prophylaxis, including all but one patient with severe hemophilia who had recently stopped taking prophylaxis. Overall outcome was good, with median HJHS score of 3.5 (range 0–30; max 140, optimum 0), median HAL score of 100 (range 62–100; max 100= optimum), and median figure 8 test 4.9 sec (range 4.2–7.4). Recent (&lt; 2 years of testing) Pettersson scores were available in only 11 patients (9 with severe, 2 with moderate hemophilia); the median score was 3 (range 0–28, max 78, optimum=0), the median physical component score of the SF36 was 53 (range 24–62), median utility (EQ5D) was 1.0 (range 0.7–1.0, optimum=1.0).
Spearman’s correlations for EQ5D, and Pearson’s correlations for other instruments are shown in the table.
Structural Functional Quality of Life HJHS Pettersson HAL Fig 8 SF 36 EQ5D HJHS 1 Pettersson 0.86 1 HAL −0.61 −0.26 1 Fig 8 0.51 0.51 −0.56 1 SF 36 −0.73 −0.41 0.81 −0.54 1 EQ5D −0.37 −0.04 0.76 −0.29 0.68 1
The HJHS scores showed good correlation with the Pettersson score and the physical component score of the SF36. Correlation with functional outcome assessment was only moderate, while the HAL showed a high correlation with both SF36 and EQ5D. In conclusion, the HJHS is a promising instrument for structural assessment, but functional assessment using the HAL showes higher correlation with Quality of Life and utility. For full assessment of results of treatment and to compare different treatment strategies, functional assessment, in addition to structural assessment, is warranted.
Assessment of Aneuploidy in Childhood Acute Lymphoblastic Leukaemia Using High Density Oligonucleotide Arrays.
Authors: Hall Andrew G. , Sulong Sarina , Harrison Christine , Minto Lynne , Bown Nick , Case Marian C. , Bailey Simon , Irving Julie 
Published: April 2020

Abstract
Chromosomal copy number (CN) is an important prognostic index in childhood acute lymphoblastic leukaemia (ALL). High hyperdiploidy (51–65 chromosomes) is associated with a good prognosis and near haploidy (23–29) a poor outcome. Conventional cytogenetics produces accurate estimates of CN in the majority of cases but sometimes cytogenetic fails or the morphology of chromosomes in ALL is too poor for accurate identification. We have used single nucleotide polymorphism arrays (SNPA) to determine CN in 86 childhood ALL patients and have compared the results with conventional cytogenetic analysis. Comparison of hybridisation intensity to a reference set of 110 normal individuals was used to derive CN estimates on 34 samples obtained during remission from ALL. Twelve were analysed using GeneChip® Human Mapping 10K Array Xba 131 and 22 using Xba 142 version 2.0 arrays (Affymetrix). Gene copy number estimates for individual SNPs were averaged across the autosomes and X chromosome. Results confirmed that SNPA gave accurate estimates of CN in samples of normal cells. Analysis by SNPA was successful in all 86 presentation leukaemic samples. The results of conventional cytogenetic analysis was available in 75, 11 having a failed cytogenetic result. Estimates of CN were identical in 34 and within 1 chromosome in a further 28. High hyperdiploidy was observed in 28 cases. In 23 of these the cytogenetic and SNPA results were concordant, while in the remaining five cases with a normal (n=2) or a failed (n=3) cytogenetic result, there was evidence from interphase FISH analysis of a hidden, non-dividing high hyperdiploid clone. One patient had a SNPA result indicative of near-haploidy (23–29 chromosomes) and another low hypodiploidy (33–39 chromosomes) with the SNPA confirming the appropriate chromosomal losses. There was marked discordance in one case of near tetraploidy (a total underestimate of 33 chromosomes) but this was not unexpected as the four copies of most chromosomes comprised no allelic imbalance. Analysis of concordance for individual chromosomes showed that, of the 1679 chromosomes compared, there was concordance of copy number in 1517 (90%). The lowest concordance was for the X chromosome (79%) and chromosome 21 (77%). This level of discordance, at least for chromosome 21, may be explained by multiple copies with no allelic imbalance. For all other chromosomes the concordance level was above 80%. We have shown that SNPA can reliably estimate CN in high hyperdiploidy and near haploid cases. This is of great potential value for genome-wide screening in cases in which cytogenetics is not available. It also provides validation of the applicability of the technique for the analysis of DNA from the majority of tumour types which do not yield metaphases.
Response to Anti CD20 Monoclonal Antibody Rituximab® and Epitope Mapping of Inhibitory Antibodies in Patients with Acquired Haemophilia.
Authors: Krause Manuela , Koenigs Christoph , Kessel Christoph , Scharrer Inge , Grossmann Ralf , Kreuz Wolfhard 
Published: April 2020

Abstract
Introduction: Acquired haemophilia (AcH) is associated with the development of polyclonal autoantibodies against FVIII, which affect directly the A2 or C2 domain of the FVIII molecule. Immunomodulatory therapy regimes to normalize FVIII levels and to eliminate the inhibitor are essential options in the treatment of patients (pts) with AcH. The aims of the present study were to investigate the response to Rituximab® and to localize the inhibitor epitopes on the FVIII domains.
Patients and Methods: In 5 pts with AcH (2 females,3 males; age: 64–81 yrs) the inhibitor titers ranged from 9 to 156 BU and the FVIII activities from &lt;1 % to 6 %. Rituximab® was administered once weekly for 4 weeks (dosage: 375 mg/m2) in combination with a prednisone therapy (1–1.5 mg/kg).
Random peptide phage display libraries were used to identify peptide sequences specific for inhibitors. The short peptide sequences allow the mapping of conformational epitopes on the surface of the FVIII molecule.
Results: In all pts (median follow up of 24 months, range: 2–49 months) we documented complete remission (CR). The median time to achieve CR was 13 weeks (range: 2–32 weeks). The time to CR was much longer (23/32 weeks) in two pts, who showed high inhibitor titers (50/188 BU) compared to pts (2/4/5 weeks) with low inhibitor titers (9/22/29 BU). One pt with a high inhibitor titer (50 BU) relapsed 19 months after CR, an other pt died in septic shock 8 weeks after the initiation of therapy with Rituximab®. No other relevant side effects were observed. In the pt with a relapse we identified target epitopes in the A1 and A2 domain of FVIII, and in the other pts (n=3) the peptide sequences mapped to epitopes on the C2 domain of FVIII.
Conclusion: The therapy with Rituximab®shows high response rates and seems to be effective in the elimination of high and low inhibitors against FVIII in pts with AcH. Limited by the small number of pts, further studies are needed to confirm whether the different target epitopes found in patients with (A1/A2) or without a relapse (C2) might have a relevance in the specific immunomodulatory therapy.
Diagnostic Workup of Patients with Acquired Von Willebrand’s Syndrome.
Authors: Tiede Andreas , Priesack Jan , Bolte Oliver , Trummer Arne , Bohlmann Katrin , Helms Wiebke , Ganser Arnold , Eisert Roswith 
Published: April 2020

Abstract
Acquired von Willebrand’s syndrome (AVWS) is a rare but probably underdiagnosed hemorrhagic disorder often associated with hematological or cardiovalvular disorders. Diagnostic workup remains challenging, particularly in patients with normal or increased von Willebrand factor antigen (Ag) and ristocetin cofactor (RCo). Here, we present a retrospective single-center study of 35 patients diagnosed with AVWS based on (i) a history of recent onset of bleeding, (ii) a negative family history of von Willebrand’s disease, and (iii) abnormal plasma VWF multimers. AVWS was associated with monoclonal gammopathy (n=11), cardiovalvular disorders (n=16), or other conditions (n=8) including myeloproliferative and autoimmune disorders. The PFA-100® screening test was inconclusive due to anemia (hematocrit &lt;30 %) or thrombocytopenia (&lt;100/nl) in 10 patients (29 %); prolonged closure times were observed using collagen/epinephrine and collagen/adrenalin in 20 of 25 (80 %) and 18 of 25 (72 %) patients, respectively. Factor VIII:C was reduced below 50 IU/dl in 7 of 35 patients (20 %). VWF Ag and RCo were reduced below 50 IU/dl in 8 patients (23 %). VWF Ag and RCo were normal or increased in all patients with cardiovalvular disease and in four of eleven patients with gammopathy. Median VWF Ag was higher in cardiovalvular disease (median 202 IU/dl, range 90 to 608) compared to gammopathy (median 31 IU/dl, range 8 to 468, p&lt;0.02 by Mann Whitney U test). Of 27 patients with normal or increased VWF Ag and RCo, 12 (44 %) had a reduced collagen binding activity (CBA) or CBA to Ag ratio &lt;0.7; 10 (37 %) had a borderline CBA ratio between 0.7 and 0.8; 5 (19 %) had a normal CBA &gt;0.8. A normal or increased VWF Ag together with a CBA ratio &gt;0.7 was observed both in patients with cardiovalvular disease (n=9), gammopathy (n=2) and other disorders (n=4). However, in all patients the largest VWF multimers were decreased (n=14) or absent (n=21). In conclusion, no single test was sufficient to detect all cases of AVWS. The PFA-100® test is of limited use in this population because of its limitation in anemia or thrombocytopenia and because of its low sensitivity. A significant number of patients present with a normal or increased VWF Ag and RCo as well as a CBA ratio &gt;0.7 emphasizing the importance of multimer analysis in all patients with suspected AVWS.
Platelet Aggregation Analysis of AMG 531 in a Randomized, Placebo-Controlled Study in Healthy Japanese Subjects.
Authors: Kumagai Yuji , Fujita Tomoe , Ozaki Machiko , Sahashi Kunihiko , Ohkura Masayuki , Ohtsu Tomoko , Arai Yoshihiro , Sonehara Yusuke , Nichol Janet 
Published: April 2020

Abstract
AMG 531 is a novel thrombopoiesis-stimulating peptibody currently being investigated for the treatment of chronic immune thrombocytopenic purpura. This study was a double-blind, phase 1 evaluation of safety (including a platelet aggregation analysis), pharmacodynamics, and pharmacokinetics of AMG 531 in healthy Japanese men. Thirty subjects were randomly assigned 4:1 (AMG 531:placebo) to each of 3 sequential dose cohorts (0.3, 1, and 2 μg/kg) of 10 subjects each. A single dose of AMG 531 or placebo was administered by subcutaneous injection, and subjects were evaluated for 6 weeks. Concentrations of 0.2 and 2 μg/mL collagen and 0.5 and 2 μmol/L adenosine phosphate were used as reagents in the platelet aggregation analysis conducted on day -1, day 2, day 15, and at the end of the study. No enhancement or reduction in platelet aggregation was detected in samples collected from subjects treated with AMG 531 compared with either pretreatment or placebo samples. Overall, adverse events were similar between the AMG 531 and placebo groups. Treatment-related adverse events (headache, “feeling hot,” migraine without aura, and/or malaise) were reported for 5 of 24 subjects treated with AMG 531. Four of 8 subjects who received the 1 μg/kg dose and 7 of 8 subjects who received the 2 μg/kg dose had platelet count increases ≥ 1.5-fold over baseline, and 3 of 8 subjects who received the 2 μg/kg dose had platelet count increases ≥ 2-fold over baseline, a pharmacodynamic effect similar to that seen in previous studies of non-Japanese subjects. Serum concentrations of AMG 531 were below the lower limit of quantification in all but 2 subjects who received the 2 μg/kg dose. In summary, platelets in subjects treated with AMG 531 functioned normally compared with pretreatment and placebo samples. AMG 531 was generally well tolerated and was effective at raising platelet counts after a single subcutaneous administration.
Deletion of Two Contiguous Genes, Platelet GPIbβ (Glycoprotein Ibβ) and Septin SEPT5, in a Boy with Bernard-Soulier Syndrome and Developmental Delay: A Possible New Contiguous Gene Syndrome.
Authors: Bartsch Ingrid , Hainmann Ina , Bläser Susanne , Pavlova Anna , Oldenburg Johannes , Busse Anja , Lanza Francois , Nurden Paquita , Superti-Furga Andrea , Zieger Barbara 
Published: April 2020

Abstract
Mutations or deletions in the genes for platelet GPIb/V/IX (consisting of GPIbα, GPIbβ, GPV, GPIX) cause the bleeding disorder Bernard-Soulier syndrome. Septins are GTP-binding proteins essential for active membrane movement such as vesicle trafficking. In non-dividing cells (i.e. platelets) septins are implicated in exocytosis. Platelets from a SEPT5 knockout mouse show altered serotonin secretion and platelet aggregation suggesting that SEPT5 is involved in secretion in platelets.
We report on a 4 year old boy born to consanguineous parents who developed several life-threatening mucosal bleeding episodes (when he suffered from upper respiratory infections) and showed developmental delay. Giant platelets were present in the blood smear, and ristocetin-induced platelet aggregation was reduced indicating that the boy suffered from Bernard-Soulier syndrome. Immune transmission electron microscopy of the platelets showed reduced expression of GPIb. PCR- and Southern analysis demonstrated that exon 1 and partly exon 2 of GPIbβ was deleted on both alleles. Further experiments showed that the deletion extended 5′ from exon 1 of GPIbβ to include the whole gene coding for SEPT5 located 5′ of GPIbβ. After bone marrow transplantation from a HLA-identical unrelated donor the boy developed no further bleeding symptoms.
Conclusions: We identified a patient with Bernard-Soulier syndrome and mental retardation who had not only a homozygous deletion of exon 1 and partly exon 2 of GPIbβ but also a deletion of the whole adjacent SEPT5 gene, possibly indicating a novel contiguous gene syndrome. Whether absence of SEPT5 contributed to particularly severe bleeding symptoms and to developmental delay remains to be proven.
Anti-Apoptotic MCL-1 Plays a Critical Role in Maintenance of Normal and Malignant Human Hematopoietic Stem Cells through FLT3 Signaling.
Authors: Yoshimoto Goichi , Miyamoto Toshihiro , Iino Tadafumi , Kikushige Yoshikane , Niiro Hiroaki , Iwasaki Hiromi , Harada Mine , Akashi Koichi 
Published: April 2020

Abstract
Mcl-1 (myeloid cell leukaemia-1) is an anti-apoptotic member of the Bcl-2 family protein, originally identified through the analysis of differentiating myeloid cells. We have recently reported that disruption of murine MCL-1 in adult murine hematopoiesis resulted in a complete deficiency of hematopoietic cells in the bone marrow, and that MCL-1 expression was activated in response to activation of c-Kit tyrosine kinase receptor signaling (Science307;1101, 2005). Here we propose that human MCL-1 also plays a critical role in maintenance of normal and malignant human hematopoiesis. The analysis of FACS-sorted human HSCs and myeloid progenitors demonstrated that, as in case of murine hematopoiesis, HSCs expressed hMCL-1 at the highest level, and that its expression gradually downregulated in committed myeloid progenitors such as CMPs and GMPs. We also found that unlike murine long-term HSCs that express c-Kit but not FLT3 receptor tyrosine kinase, human long-term HSCs express both of these receptors. Ligation of FLT3 receptor in purified human HSCs immediately induced the expression of MCL-1, suggesting that FLT3 supports human HSC survival through stimulating MCL-1 expression. We then evaluated whether hMCL-1 is expressed in leukemic stem cells. Thirty patients with acute myeloid leukemia (AML) were enrolled in this study. Within the immature CD34+ blast population, the CD34+CD38− fraction expressed higher levels of MCL-1 than those in more differentiated CD34+CD38+ fraction. In most cases, CD34+CD38− AML cells expressed 2 to 10-fold higher levels of MCL-1 transcripts as compared to normal HSCs. Interestingly, the group of samples expressed highest levels of MCL-1 constituted mostly of FLT3/internal tandem duplications (ITDs) positive AML, suggesting that constitutive activation of FLT3 signaling by FLT3 mutations might induce overexpression of MCL-1. A high level expression of MCL-1 was also found in MOLM-13 and MV4-11, FLT3/ITD positive AML cell lines. Treatment of MOLM-13 and MV4-11 with the small molecule tyrosine kinase inhibitor PKC 412, resulted in induction of apoptosis, which was associated with decreased expression of MCL-1 transcripts and proteins. Based on these data, MCL-1 might play a critical role in maintenance of normal and malignant HSCs in human at least through FLT3 signaling. Our data also suggest that constitutive activation of FLT3 signaling by FLT3 mutation might contribute to leukemic transformation through enforcing survival of AML stem cells.
Haploinsufficiency of the Platelet P2Y12 Gene in a Family with Congenital Bleeding Diathesis.
Authors: Fontana Gessica , Cattaneo Marco , Ware Jerry 
Published: April 2020

Abstract
Two related patients (sisters) with a congenital bleeding diathesis associated with a severe defect of the platelet ADP receptor, P2Y12, have been described (Cattaneo et al, ATVB2000;20:883). Platelets from these patients are severely defective in ADP-dependent aggregation and in Gi-coupled ADP-mediated responses, but are normal in Gq-coupled ADP-mediated responses. Molecular analysis of the P2Y12 gene of each sister revealed an identical single base pair (bp) deletion (378delC) occurring just beyond the coding sequence for the third transmembrane domain in P2Y12. The mutation results in a frame shift (Thr126 frame shift X34) and premature truncation of the protein (Conley et al, Blood2001; 98:43b). In the present study, we focused on a 13-year old son (GL) of one of the affected sisters. GL has not suffered spontaneous bleedings; his bleeding time is moderately prolonged (13 min). His platelets display a phenotype that is compatible with partial defect of the platelet P2Y12 receptor: abnormal aggregation and ATP secretion induced by several agonists, moderate deficiency of platelet-binding sites for [33P]2MeS-ADP, and partial impairment of inhibition of adenylate cyclase by ADP. PCR analysis of GL’s P2Y12 gene revealed only normal DNA sequence. Therefore, we hypothesized that the abnormal platelet phenotype of GL is due to haploinsufficiency of his P2Y12 gene, and, accordingly, his mother and aunt suffer from P2Y12 deficiency owing to haploinsufficiency and the previously described single base mutation in their remaining P2Y12 allele. In order to test our hypothesis, we analyzed genomic DNA from the family by Southern blotting and real time quantitative PCR. The Southern blot results suggested that GL had one P2Y12 allele only, which he inherited from his father. Our P2Y12 gene quantification strategy was based on SYBR Green detection and normalization using two reference genes with a normal copy number, ZNF 80 (3q13.31) and GPR15 (3q12.1). P2Y12 gene copy numbers, calculated by use of a comparative Ct method, revealed that GL, his mother, and aunt each contained a single intact P2Y12 allele. Thus, GL, his mother, and aunt suffer from haploinsuffiency in their P2Y12 alleles. However, GL’s remaining allele is normal, while his mother and aunt each have a remaining mutant allele. This explains the differences in clinical phenotypes between GL and his mother and aunt. The quantitative PCR result, supported by Southern blotting results, supports our hypothesis and illustrates the platelet phenotype associated with P2Y12 haploinsufficiency.
14-3-3 Integrates Pro-Survival Signals in Hematopoietic Cells Transformed by Diverse Leukemogenic Tyrosine Kinases, and Represents a Common Target.
Authors: Dong Shaozhong , Kang Sumin , Gu Ting-lei , Kardar Sean , Lonial Sagar , Khoury H. Jean , Khuri Fadlo , Fu Haian , Chen Jing 
Published: April 2020

Abstract
Constitutively activated tyrosine kinases associated with recurrent chromosomal abnormalities play an essential role in the pathogenesis and disease progression of a variety of hematopoietic malignancies. Selective tyrosine kinase inhibitors such as imatinib are effective in treating some forms of leukemia such as t(9;22) CML associated with expression of BCR-ABL fusion tyrosine kinase. However, they are not curative and clinical resistance may develop, prompting the design of alternate and/or complementary therapeutic strategies. To better understand the signaling properties of constitutively activated tyrosine kinases associated with different hematopoietic malignancies, we examined whether BCR-ABL, FLT3-ITD, NPM-ALK, TEL-PDGFbetaR, TEL-FGFR3 and ZNF198-FGFR1 activate the same set of signaling pathways. We found that they all activated AKT and MAPK signaling pathways. Activated AKT resulted in phosphorylation of FOXO3a at Thr-32 but not BAD at Ser-136, whereas activated MAPK led to phosphorylation of BAD at Ser-112. These phosphorylated residues subsequently sequestered the pro-apoptotic FOXO3a and BAD to 14-3-3, suggesting that 14-3-3 integrates pro-survival signals from AKT and MAPK pathways. We utilized a peptide-based 14-3-3 competitive antagonist, R18 to disrupt 14-3-3/ligand association. Expression of R18 effectively induced apoptosis in hematopoietic Ba/F3 cells transformed by these tyrosine kinases with significantly enhanced sensitivity compared to the control Ba/F3 cells. Moreover, doxycycline-induced expression of R18 significantly attenuated the disease latency and penetrance in mice induced by intravenous injection of representative ZNF198-FGFR1-transformed Ba/F3 cells. Co-immunoprecipitation experiments indicate that induced R18 expression disrupted interaction between 14-3-3 and FOXO3a, but not 14-3-3/BAD association. R18 induced apoptosis by rescuing the nuclear localization of FOXO3a and up-regulating FOXO3a transcription targets Bim and p27 in cells expressing ZNF198-FGFR1. Furthermore, fluorescent confocal microscopy revealed that expression of R18 generally resumed FOXO3a nuclear localization in cells transformed by the spectrum of diverse leukemogenic tyrosine kinases. Together, these data support a model that 14-3-3 functions as a general integrator of pro-survival signals in hematopoietic transformation induced by diverse leukemogenic fusion/mutant tyrosine kinases. Disrupting 14-3-3/ligand association may be a common and effective therapeutic strategy for hematopoietic neoplasms associated with these tyrosine kinases.