bluuebunny/crossref_metadata_2025_split huggingface dataset:
Autocrine Signaling by Osteopontin Regulates Actin Remodeling in Human Primary Erythroid Progenitors.
Authors: Wickrema Amittha , Zhou Ying , Reynolds Jessica , Ulaszek Jodie , Satgurunathan Nilesh , van Besien Koen , Kang Jeong-Ah 
Published: October 2019

Abstract
Osteopontin (OPN) is a secreted protein shown to have multiple functions depending on cell type and context. The cell types known to express OPN are typically assocoated with immune and /or bone remodeling functions. Here we demonstrate that OPN is also expressed and secreted by primary human erythroid progenitors and that it is involved in actin remodeling in erythroid cells. We initially identified OPN as an important cytokine in erythroid differentiation by screening a cytokine/receptor cDNA microarray. Subsequently using a culture system of highly purified primary erythroid progenitors (sorted for glycophorin A) we examined OPN mRNA and protein expression during terminal erythroid differentiation. We demonstrated that OPN mRNA is expressed throughout differentiation and that OPN protein is also detectable at basophilic, polychromatic and orthochromatic stages during the maturation program. Furthermore by ELISA we were able to detect secretion of OPN at each of the major stages of differentiation. In addition we show that CD44 and integrin alpha 4, two of the receptors for OPN are highly expressed in these cells. Stimulation of these cells with rOPN resulted in phosphorylation of several proteins one of which was identified as the heat shock protein-27 (Hsp27). In subsequent studies we were able to demonstrate that OPN regulates phosphorylation of Hsp27 at serine-78 site through the p38 MAP kinase pathway. We then effectively knocked down the expression of OPN by siRNA methodology, which resulted in formation of distinct F-actin bundles in the perimembrane region of the cell. By contrast in cells transfected with control siRNA maintained F-actin in a punctate appearance. Altogether our studies suggest a critical signaling function for OPN in regulation of atleast one of the erythrocyte cytoskeletal elements during erythroid differentiation.
Mapping of the Glycolytic Enzyme Binding Sites on Human Erythrocyte Membrane Band 3.
Authors: Chu Haiyan , Low Philip S. 
Published: October 2019

Abstract
Glycolytic enzymes (GEs) including aldolase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphofructokinase (PFK), pyruvate kinase (PK) and lactate dehydrogenase (LDH) are known to associate with the inner surface of the human erythrocyte membrane. Previous studies have suggested that the N-terminus of the cytoplasmic domain of band 3 (cdb3) constitutes the likely binding site for aldolase, GAPDH and PFK, however, the membrane docking sites for PK and LDH have not been elucidated. In this study, we demonstrate that PK and LDH exhibit no affinity for band 3, regardless of whether the association is measured by co-immunoprecipitation assay, binding competition studies, or catalytic inhibition analyses. We further find that the binding sites for GAPDH, aldolase and PFK on band 3 are distinct but partially overlapping, as evidenced by the fact that: 1) deletion of residues 1–11 of cdb3 eliminates the binding of aldolase, but not PFK or GAPDH, 2) fusion of thioredoxin (Trx) to the N-terminus of residues cdb3 blocks aldolase binding, but not the association of GAPDH or PFK, 3) deletion of sequences 1–50, 1–40, 1–31, or 1–23 of cdb3 blocks cdb3 association with all three GEs, whereas deletion of residues 12-23 only abrogates aldolase binding (while reducing the affinity of PFK and GAPDH), 4) the presence of both sequences, 6–DDYED-10 and 19-EEYED-23, are necessary for cdb3 association with aldolase, whereas the presence of either sequence alone (especially 19-EEYED-23) is sufficient to maintain association with GAPDH and PFK, 5) mutation of all of the acidic residues in the above two sequences to their corresponding amides (E→Q and D→N) results in loss of affinity for all GEs. Because i) kidney cdb3 (which lacks residues 1-65 of cdb3) shows no affinity for any of the GEs, ii) residues 1–55 of cdb3 show near normal affinity for aldolase, GAPDH and PFK, and iii) GAPDH, aldolase, and PFK all compete with each other for cdb3, we conclude that the binding sites for aldolase, GAPDH and PFK are all located within the first 23 residues of cdb3, with the docking site for aldolase likely residing somewhat more N-terminal than the binding sites for GAPDH and PFK. Finally, because each band 3 monomer contains three homologous sequences (6-DDYED-10, 19-EEYED-23 and 902-DEYDE-906) that are found in other proteins that bind GEs (e.g. actin, β-tubulin, troponin T), and since the first two of these sequences were shown to be required for cdb3 binding, the question naturally arose whether different GEs might also associate with residues 902-DEYDE-906 at the extreme C-terminus of band 3. Similar binding studies demonstrate that such an interaction does not occur. Thus, our data show that GAPDH, aldolase, and PFK all bind near the N-terminus of band 3 and that PK and LDH must dock somewhere else on the red cell membrane.
Myocardial T2* Is Not Affected by the Structural Effects of Myocardial Fibrosis, Ageing or Impaired Left Ventricular Function.
Authors: Kirk Paul , Pennell Dudley J. 
Published: October 2019

Abstract
Background
The myocardial T2* technique has been validated as a reproducible non-invasive measurement of myocardial iron load and is now widely used for measurement of myocardial iron in iron overload diseases such as thalassaemia. The reduction in myocardial T2* seen in iron overload conditions is substantially greater than is seen in any other clinical circumstance, but there has been no direct comparison of myocardial T2* in normals and other conditions such as increasing age, myocardial infarction or impairment in left ventricular function. We aimed therefore to compare the findings in patients affected by these conditions with normals.
Method
A total of 38 patients in total were scanned using the myocardial T2* technique. Fifteen patients had normal hearts, 18 had impaired LV function and 6 had chronic myocardial infarction affecting the anteroseptal wall, where myocardial T2* measurements are normally made.
Results
The mean myocardial T2* in normals was 36.0 +/− 6.4 ms, yielding a lower limit of normal of 23ms. In patients with impaired LV function, the mean myocardial T2* was 39.0 +/− 11.7ms (p= 0.37 vs normals). In patients with anteroseptal myocardial infarction, the mean myocardial T2* was 34.7ms +/− 3.9ms (p= 0.64 vs normals). The frequency distribution of the myocardial T2* values are shown in figure 1. These approximate to normal, and are very similar in distribution. In addition, the age distribution of myocardial T2* in the 15 normals is shown in figure 2. There was no significant relation between myocardial T2* and age (r2 = 0.066, p=0.82).
Conclusion
There is no significant reduction in myocardial T2* associated with fibrosis from chronic myocardial infarction, impairment of left ventricular function, or increasing age. This suggests that structural changes associated with remodelling, infarction and fibrosis, and ageing do not have significant effects on the absolute measure of myocardial T2*, and in particular do not cause a reduction below 20ms as is seen in myocardial overload conditions. Thus these date suggest that myocardial T2* is robust to these structural alterations, and that myocardial iron overload can be ascertained from reduced myocardial T2* values, in a similar manner to that which can be achieved in normals.
Figure 1 Figure 1. Figure 2 Figure 2.
Assessment of Serum Free Light Chain Assay for Screening for Plasma Cell Disorders.
Authors: Smith David E. , Abadie Jude , Bankson Daniel , Mead Graham 
Published: October 2019

Abstract
Introduction and Methods: The purpose of this study was to evaluate the serum free light chain (FLC) assay in its ability to improve performance of protocols designed to screen for plasma cell disorders. We measured M-protein levels using serum protein electrophoresis (SPEP) in 312 consecutive patients being screened for plasma cell disorders at the Veterans Administration Medical Center - Puget Sound. The serum kappa and lambda free light chain levels were quantitated using the serum FLC assay in these same patients. The kappa/lambda ratio was calculated using the free kappa and free lambda results from the serum FLC assay.
Results: SPEP results indicated the presence of a possible monoclonal gammopathy in 77 of the 312 patients in this study. In this group of 77 patients, a plasma cell disorder was diagnosed in 27 of them. The serum FLC assay showed an abnormal kappa/lambda ratio in 20 of these 77 patients, all 20 of whom were diagnosed with multiple myeloma. In the group of 235 patients with normal SPEP results, 17 were found to have an abnormal kappa/lambda ratio. Of these 17 patients, 15 were diagnosed with multiple myeloma, one with lymphoma, and one with bladder cancer.
Conclusions: Because a number of disorders and diseases can increase production of immunoglobulins, there were a significant number of false positives in the SPEP results. At the same time, there were also several false negative SPEP results. The number of both false positives and false negatives was smaller for the serum FLC assay. Further, use of SPEP and the serum FLC assay together resulted in significantly improved sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). (See Table 1.) These results indicate an important role for the serum FLC assay in screening for monoclonal gammopathies.
Table 1. Performance of SPEP, sFLC, and both assays in screening for plasma cell disorders SPEP Alone sFLC Alone Both SPEP and sFLC Sensitivity 64% 88% 100% Specificity 81% 98% 99% Positive Predictive Value 35% 88% 89% Negative Predictive Value 94% 98% 100%
Upshaw-Schulman Syndrome: A Masqueraded Thrombocytopenia during Pregnancy.
Authors: Matsuyama Tomomi , Matsumoto Masanori , Kato Seiji , Akiyama Nobu , Tomiyama Junji , Natori Kazuhiko , Kuraishi Yasunobu , Imamura Yutaka , Inoue Nobumasa , Kokame Koichi , Miyata Toshiyuki , Fujimura Yoshihiro 
Published: October 2019

Abstract
Upshaw-Schulman syndrome (USS) is a congenital thrombotic thrombocytopenic purpura (TTP) characterized by a deficiency of ADAMTS13 activity due to its mutations. More than half of USS patients have an episode of severe jaundice during the newborn period and require exchange blood transfusions. Beyond this period, however, the clinical manifestations of USS vary significantly from patient to patient, and do not usually include Moschowitz’s pentad, a hallmark of TTP. However, a consistent clinical sign in these patients is an occasional or chronic thrombocytopenia that is enhanced by precipitating factors such as viral infections. Therefore, USS patients are often incorrectly diagnosed with idiopathic thrombocytopenic purpura (ITP) or Evans syndrome during childhood, and the underlying ADAMTS13 gene defects are overlooked.
Here, we report on 7 female USS patients belonging to 4 different families, including 3 pairs of siblings who were uniformly diagnosed by episodes of thrombocytopenia or prevalence of TTP at 5~6 months of pregnancy. Plasma ADAMTS13 activity was measured with a highly sensitive ELISA using a monoclonal antibody directed against the VWF-A2 domain developed in our laboratory. The ADAMTS13 activity and gene mutations in these patients are described:
Family K: an elder sister K1 (activity 0.69%; gene mutation Y304C/G525D) and a younger sister K2 (activity <0.5%; gene mutation Y304C/G525D), Family L: an elder sister L1 (activity 1.3%; gene mutation 372insGT/Q1302X) and a younger sister L2 (activity 0.96%; gene mutation 372insGT/Q1302X), Family M: an elder sister M1 (activity <0.5%; gene mutation R193W/R349C) and a younger sister M2 (activity <0.5%; gene mutation R193W/R349C), and Family O: a patient (activity <0.5%;gene mutation I178T/Q929X). Figure indicates family pedigree of USS involved in this study.
Fig. Family pedigree of Upshaw-Schulman syndrome Fig. Family pedigree of Upshaw-Schulman syndrome
Interaction of beta2-Glycoprotein I with Members of the Low Density Lipoprotein Receptor Family.
Authors: van Lummel Menno , Pennings Maarten T.T. , Derksen Ronald H.W.M. , Westein Erik , Lenting Peter J. , de Groot Philip G. 
Published: October 2019

Abstract
The antiphospholipid syndrome (APS) is a non-inflammatory disease characterized by arterial and/or venous thrombosis and/or pregnancy morbidity in the presence of auto-antibodies that recognize beta2-glycoprotein I (beta2GPI) bound to cardiolipin. We have previously demonstrated that dimerization of beta2GPI by auto-antibodies induces platelet activation. This effect has been shown to be mediated by apoER2′. Here, we show that dimeric beta2GPI, and not plasma beta2GPI interacts with three different LDL-receptor family members; apolipoprotein E receptor 2′ (apoER2′), the low LDL-receptor related protein (LRP) and megalin. Interaction between dimeric beta2GPI and apoER2′ was best described with a one-site binding model (KD(app) 25 nM), whereas for LRP and megalin the interaction with dimeric beta2GPI was best described with a two-site binding model, representing a high- (3.1 nM) and a low-affinity site (192.1 and 241.2 nM, respectively). Binding kinetics of a dimeric beta2GPI mutant, that binds poorly to anionic phospholipids were similar to the binding kinetics of full length dimeric beta2GPI for all three LDL-receptor family members. Upon deletion of domain I or domain II of dimeric beta2GPI no changes in the binding kinetics were observed. Deletion of domain V however, significantly decreased the affinity for apoER2′, LRP and megalin. In conclusion, our data show that dimeric beta2GPI can interact with different LDL-receptor family members. This interaction is dependent on a recognition site in domain V of beta2GPI, which does not overlap with the phospholipid-binding site.
A Role of the Protein Tyrosine Kinase, Syk in Complement-Mediated Phagocytosis: A Mechanism on the Engulfment of Pathogen.
Authors: Tohyama Yumi , Shi Yuhong , Tohyama Kaoru , Yamamura Hirohei 
Published: October 2019

Abstract
A protein-tyrosine kinase, Syk plays a central role in Fc-gamma receptor-mediated phagocytosis in the adaptive immune system. Here we show that Syk also makes an essential role in compliment-mediated phagocytosis in innate immunity. For this purpose, HL60 cells induced to macrophage-like with vitamin D3 and TPA and C3bi-opsonized zymosan were used as the pathogen-phagocyte system. Time-lapse microscopic observation revealed that C3bi-opsonized zymosan particles were promptly attached to and sucked into the cells but non-opsonized zymosan was not, and therefore it was indicated that our phagocytosis system is mediated by the complement receptor 3 (CR3). Syk was tyrosine-phosphorylated by the treatment with C3bi-opsonized zymosan and marked accumulation of Syk was detected around the newly forming phagosomes. To further investigate the effect of Syk on complement-mediated phagocytosis, Syk-siRNA or dominant-negative Syk (DN-Syk) was introduced into HL60 cells. Quantitative analysis by flow cytometry showed that phagocytosis was severely impaired in DN-Syk or Syk-siRNA expressing HL60 cells. To determine whether the impaired capture of zymosan is due to reduced binding to CR3 or reduced engulfment after binding, the cells were treated with a quenching reagent which discriminates fluorescent zymosan particles between inside and outside the cells. In the parental HL60 cells most of the attached zymosan particles were inside the cells, while few particles were ingested in the case of DN-Syk or Syk-siRNA expressing HL60 cells. These data indicated that Syk is required for engulfment of C3bi-opsonized zymosan but not for attachment of zymosan to CR3. To clarify the molecular mechanism how Syk affects the phagosome formation and its engulfment, the behavior of F-actin was searched microscopically. At the early stage of complement-mediated phagocytosis, marked accumulation of F-actin occurred around each phagosome and was followed by rapid depolymerization in the parental HL60 cells. In contrast, phagosomes surrounded by the accumulated actin were significantly decreased in DN-Syk or Syk-siRNA expresssing cells. These results indicate that Syk promotes actin assembly around phagosomes prior to engulfment at the early stage of the phagocytosis. As for the analysis of the downstream pathway of Syk in our phagocytosis system, RhoA-signaling was investigated. Vav-RhoA pathway was activated in HL60 cells but transfer of DN-Syk or Syk-siRNA into HL60 cells reduced the activation of Vav-RhoA signaling. Therefore Syk may act as an activator of the RhoA-pathway in phagocytosis.
Our results demonstrate that Syk is required for complement-mediated phagocytosis by regulating both actin accumulation around phagosomes and RhoA-signaling harmonically and finally leads to phagosome formation and its engulfment.
SFCE (Societe Francaise des Cancers de l’Enfant) Recommendations for the Management of Tumor Lysis Syndrome (TLS): A Validation Survey.
Authors: Bertrand Yves , Auvrignon Anne , Nelken Brigitte , Mialoux Valérie , Mechinaud Francoise , Patte Catherine , Plantaz Dominique , Le Guyader Nathalie , Urbieta Marisol , Perel Yves , Baruchel9 Andre , Leverger Guy 
Published: October 2019

Abstract
TLS is a metabolic complication of haematological malignancies, which can translate in renal impairment. As previously described (Goldman S.C. et al,Blood2001; 972998–3003 and Pui C.H.et al., J.Clin.Oncol.2001;19,697–704), some patients are at High Risk (HR) of developing TLS. Rasburicase (R) a recombinant urate oxidase which transforms uric acid into a highly soluble compound, allantoin, more easily excreted by the kidneys, is highly effective in the prevention and treatment of TLS. SFCE has made recommendations for the management of TLS in children and developed a prospective survey in order to validate these recommendations.
Patients and Methods: HR pts were defined as: Acute Lymphoblastic (ALL) or Myeloblastic (AML) Leukaemia with initial leucocytes counts of at least 50x109/L, B cell ALL, Stage III and IV T or B Non Hodgkin Lymphoma (NHL), any leukaemia or NHL with a plasma uric acid concentration (U) of at least 300mmoles/L if ≤ 10 years old or 350mmoles/L if ≥ 10 years old, or any disease with serum creatinine (C) or lactate deshydrogenase concentration (LDH) exceeding twice the upper limit of normal (N), hyperphosphatemia (P)≥ 2 mmoles/L. These pts received hydration (3L/m2) ± alkalinization and R 0,20 mg/kg/d x 5 days. If C >1,5 N or P> 3mmoles/L or U ≥ 200mmoles/l, R administration is prolonged until normalization. Low risk (LR) pts, defined as not HR, received hydration 3L/m2 and R 0,20 mg/kg/for only one day. R could be prolonged according to the same criteria as HR pts.
Results: Between May and December 2004, 174 patients including 91 boys and 83 girls (median age 5 years) were treated according to these recommendations, in 8 centers in France. Initial diagnosis was LA in141 and NHL in 33. 143 patients (82%) were classified HR and 31 LR (18%). In the HR group, the patients received a median of 5 days of treatment with R (1–12) and in the LR group a median of 1 day (0–5). 25% of HR patients required treatment prolongation, while the LR group treatment prolongation was needed for 1 patient (3%). No complication of TLS was observed.
Conclusions: these results show that the risk classification as established by the SFCE was accurate and that the treatment guidelines was effective for the control of hyperuricemia and TLS.
Molecular Analysis of β-Thalassemia Cases in Poland.
Authors: Zdebska Ewa , Burzynska Beata , Mendek-Czajkowska Ewa , Maciag Malgorzata , Krawcewicz Angelika , Mokras Urszula , Adamowicz-Salach Anna , Koscielak Jerzy 
Published: October 2019

Abstract
β-thalassemia is a heterogenous, inherited disease resulting from reduced or absent synthesis of β-globin chain of haemoglobin. This disorder is very common in the Mediterranean, Middle Eastern, African and South-East Asian populations. The aim of the present investigation was to verify a common view that thalassemia in Poland is a very rare disease. 600 patients (270 men and 330 women), aged 2–85 years, with microcytosis and no evidence of iron deficiency, were examined for β-thalassemia. Hemoglobin A2 was increased in 106 patients. In 48 patients there was also an elevation of hemoglobulin F and in 42 patients of serum bilirubin.
All 106 patients were examined for 8 common Mediterranean mutations. 7 different mutations were detected in 46 heterozygous patients (number of patients wit a particular mutations are in square brackets): IVS1-6(T>C) [15]; IVS2-745(C>G)[14]; IVS2-1(G>A) [10]; IVS1-1(G>A) [2]; CD6-A [2]; CD39(C>T) [2]; IVS1-110(G>A) [1].
We also discuss the effect of these mutations on the expression level of the β-globin gene. Frequencies of individual mutations in Poland were different from those encountered in Mediterranean and some Centrak European countries. We also detect two novel mutations:
IVS-1-1(G>A) G^GTTGGT>AGATTGGT and 5′UTR; +33(C>T). The first novel mutation significantly decreased the mRNA level of the β-globin gene. The second mutation seems to be silent and does not affect β-globin synthesis. In this case, this particular mutation occurs with IVS-1-6(G>C) mutation in compound the heterozygotes stage.