bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ex Vivo Expansion of Cord Blood Hematopoietic Stem and Progenitor Cells in a Wharton’s Jelly and Bone Marrow Stromal Cell Based Co-Culture System
Authors: Li Dandan , Aljitawi Omar S. , Hopkins Richard A. 
Published: October 2019

Abstract
Umbilical cord blood (UCB) is of great value by providing transplantable hematopoietic stem and progenitor cells (HSPCs). Compared with HSPCs from adult bone marrow and periferial blood, UCB cells are more primitive with higher proliferation ability, and UCB HSPC transplantation requires less HLA matching. The major problems in UCB transplantation is the limited number of transplantable cells in each unit which are often insufficient for transplantation in adults
In order to elucidate the effects of the main components of bone marrow on cord blood CD34+ expansion, freshly enriched cord blood CD34+ cells were cultured in contact with bone marrow stromal cell (BM-MSC) monolayer, BM-MSCs pre-seeded in decellularized Wharton's jelly matrix (DWJM), and in DWJM alone, as well as separated by a transwell preventing the physical contact between CD34+ cells and BM-MSCs in a medium supplemented with a cytokine cocktail including Flt-3 igand, stem cell factor, and thrombopoietin. Expansion patterns were analyzed by CD34+ and CD45+ expansion, as well as colony-forming unit (CFU) assay. We found that the number of CFU increases significantly in cord blood CD34+ cells co-cultured with BM-MSCs in DWJM compared to that of control. Particularly, the increase in CFU-GEMM, BFU-E and CFU-GM number is more significant compared to other colonies. Importantly, DWJM alone is not able to increase CFU numbers compared to that of the control indicating there is a synergistic effect between DWJM and BM-MSCs on cell stemness. Surprisingly, BM-MSCs in DWJM also increses CD34+ cell expansion by 2 to 3 fold after one week culture, presumably from enhanced ablilty of self-renewal from CD34+ cells. Therefore, our data suggest DWJM synergizes with BM-MSCs to increase CD34 cell stemness, which can potentially be used in the clinic therapy.

Disclosures
No relevant conflicts of interest to declare.
A Genetic Variant in the CD53 Gene Is Associated with Clinical Outcomes after Unrelated Bone Marrow Transplantation
Authors: Espinoza Luis Jorge , Matsuo Keitaro , Toda Saki , Morishima Yasuo , Onizuka Makoto , Kashiwase Koichi , Fukuda Takahiro , Kodera Yoshihisa , Nakao Shinji , Takami Akiyoshi 
Published: October 2019

Abstract
Background: CD53, a member of the tetraspanin family of proteins, broadly expressed in hematopoietic cells, is implicated in leukocyte activation and cell survival responses to stress conditions. In a recent genome linkage scan, the single nucleotide variation, rs6679497 (C>G), in the intron of the CD53 gene was identified as an important regulator of TNF release from innate immune cells in response to inflammatory stimuli. (Bos SD: Eur J Hum Genet. 2010). Tumor necrosis factor alpha (TNF-ƒ¿) which is released pretransplant due to irradiation and cytotoxic preconditioning regimens, has been implicated in transplant complications.
Methods: To assess the impact of rs6679497 variation of the CD53 gene, on transplant outcomes, we evaluated 322 of patients with hematologic malignancies transplanted through the Japan Marrow Donor Program. The functional relevance of the rs6679497 variation was investigated in a whole blood culture system using samples from healthy individuals. To investigate the effect of rs6679497 on mRNA transcription activity we performed allele-specific quantitative RT-PCR with a TaqMan probe of rs6679497 using cDNA from in vitro activated leukocytes from healthy donors with heterozygous genotypes.
Results: There was a significantly lower overall survival in the recipients with the GG genotype in the univariate analysis (29% vs. 53% at 5-y; P=0.01) and multivariate analysis (relative risk=1.97; 95% confidence interval, 1.22-3.18; P=0.01, Figure 1). The donors s6679497 genotypes did not significant influence the transplant outcomes. In the allele-specific quantitative PCR, the mean ratio (G allele transcripts vs. C allele transcripts) was 1.5 which is higher than that of DNA amplicons, suggesting that the G allele has higher transcriptional activity than the C allele. Using the TF-SEARCH online tool, which predict putative transcription factor binding sites, the genomic region containing the G allele of s6679497 was found to create a novel consensus sequence corresponding to the putative binding element of GATA-1. In vitro stimulated leucocytes from healthy individuals possessing the G allele secreted higher levels of TNF-ƒ¿ than those without the G allele. Consistent with these results, flow cytometry studies revealed that monocytes from donors possessing the G allele express higher levels of CD53 on their surface (p=0.02) which may account for their higher TNF-ƒ¿ secretion. These findings substantiate the functional relevance of the rs6679497 variation, and indicate that the increased TNF-ƒ¿ secretion by individuals with the G allele likely account for their worse OS.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Meta-Analysis Evaluating Efficacy of High-Dose Therapy and Autologous Hematopoietic Cell Transplantation As Front-Line Consolidation in Chronic Lymphocytic Leukemia: Results Do Not Justify This Treatment Approach
Authors: Reljic Tea , Kumar Ambuj , Djulbegovic Benjamin , Kharfan-Dabaja Mohamed A 
Published: October 2019

Abstract
Background: High-dose therapy (HDT) followed by autologous hematopoietic cell transplantation (auto-HCT) has been offered to patients with chronic lymphocytic leukemia (CLL), both as front-line consolidation and in the relapsed setting. Uncertainty remains in regards to the role of HDT in the front-line consolidation setting in CLL. Accordingly, we performed a systematic review and meta-analysis aiming at evaluating the totality of evidence pertaining to the efficacy (or lack thereof) of HDT and auto-HCT as front-line consolidation in subjects with CLL.
Materials and methods: A total of 475 references were identified through a systematic search of PUBMED/MEDLINE and Cochrane through June 26, 2014. Only four randomized controlled trials which included a total of 600 subjects were eligible for inclusion in this meta-analysis. Data was meta-analyzed for benefits: progression-free survival (PFS) (data from 2 studies (total n=178)), event-free survival (EFS) (data from 2 studies (total n=422)), PFS or EFS (data from 4 studies (total n=600)), and overall survival (OS) (data from 4 studies (total n=600)); and harms: treatment related mortality (TRM) (data from 2 studies (total n=276)), development of any secondary malignancy ((data from 4 studies (total n=581)), or development of secondary MDS/AML in particular (data from 4 studies (total n=581)).
Results: These results are reported in accordance with Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines. Four studies enrolled a total of 301 subjects in the HDT/auto-HCT arm and 299 subjects in the control arm. Offering front-line HDT/auto-HCT did not improve PFS [Hazard ratio (HR)=0.70 (95%CI= 0.32, 1.52), p=0.37] or OS [HR=0.91 (95%CI= 0.62, 1.33), p=0.64]. An advantage favoring HDT/auto-HCT was observed in terms of EFS (HR=0.46 (95%CI= 0.26, 0.83), p=0.01] or when analysis included PFS or EFS [HR=0.54 (95%CI= 0.35, 0.82), p=0.004]. Moreover, HDT/auto-HCT did not result in higher TRM (Risk ratio (RR)=1.32 (95%CI= 0.43, 4.06), p=0.63]. When analyzing development of any secondary malignancy, no difference was observed whether subjects were offered HDT/auto-HCT or not [RR=1.06 (95%CI=0.55, 2.05), p=0.86]. A two-fold higher incidence of secondary MDS/AML, albeit not statistically significant, was observed with the HDT/auto-HCT approach [RR=1.95 (95%CI=0.60, 6.34), p=0.27].
Conclusion: Offering HDT/auto-HCT as front-line consolidation in patients with CLL does not improve OS. At the present time, this approach should be offered only in the context of a clinical trial.

Disclosures
No relevant conflicts of interest to declare.
A Study of Phenotypic Heterogeneity in Hb E-Beta Thalassemia and Its Correlation with Hb E Level
Authors: Bhattyacharyya Maitryee , Bagchi Basab , Kumar Meet 
Published: October 2019

Abstract

INTRODUCTION: Hb E-beta thalssemia has variable clinical phenotype, varying from NTDT to severe transfusion dependent thalassemia. Factors influencing the severity include HbF level, type of beta mutation, co-inheritance of alpha mutation (including triplication and quadruplication), and various polymorphisms. We evaluate the possible correlation between HbE levels and clinical severity in patients in Hb E-beta thalassemia.
AIMS AND OBJECTIVES: To study the phenotypic heterogeneity of Hb E-beta thalassemia patients with respect to Hb E levels.
MATERIALS AND METHODS: Patients >18years were enrolled and evaluated for clinical phenotype as per Mahidol score (age of onset, age of first transfusion, requirement for transfusion, spleen size, growth retardation and steady-state hemoglobin), and classified into mild, moderate and severe. Further biochemical data was evaluated including serum ferritin. The clinical phenotype severity was correlated with HbE and HbF levels. Patients on disease modifying drugs (eg hydroxyurea) were excluded.
RESULTS: A total of 120 patients of Hb E-beta thalassemia were enrolled with M:E = 1.1:1. 65.8% patients were 18-30 years age range, while 5.8% were above 50years of age. As per Mohidol score, 39.2% (47) patients were mild, 45.8% (55) were moderate and 15% (18) were severe phenotype. Observations amongst patients of different clinical severity are as shown in Table 1. Abstract 1365. Table 1.CharacterMildModerateSevereRangeP valueNumber475518Baseline Hb (g/dl)7.9+-0.96.4+-1.16.2+-0.73.8-10.30.000Age at presentation (years)18.3+-8.7613.1+-8.03.16+-2.081-440.000Age at 1st transfusion (years)22.1+-11.415.1+-8.763.9+-1.91-600.000Freq of transfusion (annual)2.5+-4.512.2+-7.619.3+-11.40-480.000Spleen size (cms)5.6+-2.47.8+-2.910.4+-3.40-150.000Serum ferritin (ng/ml)792.5+-534.61206.5+-523.91570.5+-581.264.2-2924.20.000
Hb E levels of the patients ranged from 28% to 83.5%, and HbF levels ranged from 3.1% to 49.4%. On statistical analysis, difference in HbE levels was significant between severe vs mild and intermediate groups (p=0.0005 and 0.012 respectively), but not amongst mild vs intermediate groups (p=0.21). Also, difference in HbF levels was significant between mild vs moderate and severe groups (p=0.007 and 0.004 respectively), but not between moderate vs severe groups (p=0.26). Regression analysis showed HbF and HbE to be independent parameters of severity and both have negative correlation with disease severity.
CONCLUSION: HbE is an independent parameter of severity and correlated negatively with disease severity in HbE-beta thalassemia.

Disclosures
No relevant conflicts of interest to declare.
Functional Analysis of Phage Display Selected Single-Chain Variable Antibody Fragments (scFvs) Specific for Anti-FVIII Antibodies
Authors: Brettschneider Kerstin , Naumann Anja , Neimanis Sonja , Kahle Joerg , Heller Christine , Klingebiel Thomas , Schwabe Dirk , Koenigs Christoph 
Published: October 2019

Abstract
The development of inhibitory antibodies against coagulation factor VIII (FVIII) is currently the most serious complication for hemophilia A patients that undergo FVIII replacement therapy. In addition, non-hemophilia A patients can spontaneously develop inhibitory auto-antibodies to FVIII, which results in acquired haemophilia A. The control of the allo- or autoimmune response to FVIII apparently includes the elicitation of anti-idiotypic antibodies. In this study the capacity of anti-idiotypic single-chain variable antibody fragments (scFvs) for neutralization of inhibitory anti-FVIII antibodies (FVIII inhibitors) was evaluated in vitro and in vivo.
Anti-idiotypic scFvs were selected from phage-displayed libraries against murine monoclonal FVIII-specific inhibitors. As the majority of inhibitory antibodies is directed against the A2 or C2 domain of FVIII, strongly inhibitory A2- and C2-specific antibodies served as targets. Selected scFvs were expressed as scFv-Fc fusion proteins.
Analysis of the scFv-Fcs by ELISA confirmed the specific binding to the cognate targets and binding studies via surface plasmon resonance revealed high affinities within the nanomolar range. Further characterization showed that binding of inhibitors to immobilized FVIII was blocked by specific scFv-Fcs in vitro. The ability of scFv-Fcs to neutralize their corresponding inhibitors was analyzed in a functional clotting assay. By adding scFv-Fcs to plasma spiked with inhibitors, FVIII activity was restored to 80% in a concentration dependent manner.
FVIII knockout mice served as model organism for testing the capacity of scFv-Fcs to restore coagulation in vivo. Subsequent injection of FVIII following the injection of the inhibitors resulted in a largely reduced FVIII activity. However, FVIII activity was recovered in a concentration dependent manner by adding cognate anti-idiotypes. The scFv-Fcs were either preincubated with the corresponding inhibitor or added to the FVIII mixture without preincubation. The latter represents an adaption to a therapeutic setting.
In conclusion, phage display selected anti-idiotypic scFvs are able to bind and effectively neutralize their target inhibitors in vitro and in vivo. Based on these promising results the potential of anti-idiotypic scFvs for the development of specific cell based immunotherapies for hemophilia A patients with inhibitors is currently under investigation.

Disclosures
No relevant conflicts of interest to declare.
CHRONIC NEUTROPENIA
Authors: HATTERSLEY PAUL G. 
Published: October 2019

Abstract
A case of severe chronic neutropenia without splenomegaly is reported. The disorder was characterized by chronic fatigue and many pyogenic infections, and by a persistently low neutrophile count. The marrow was cellular, with a defect in maturation in both granulocytic and megakaryocytic series. There was no improvement on the usual types of medical therapy, and splenectomy, while it was followed by transient thrombocytosis, likewise failed to induce a remission.
It is suggested that in the consideration of therapy for severe neutropenia, a lack of splenomegaly should be considered a contraindication to splenectomy.
CHRONIC LEG ULCER IN DISEASES OF THE BLOOD
Authors: GENDEL BENJAMIN R. 
Published: October 2019

Abstract
The literature pertaining to the association of chronic leg ulcer with diseases of the blood other than sickle cell anemia has been reviewed.
Two patients with this association have been presented. One patient had a hemolytic anemia and the other a pancytopenia (secondary hypersplenism) in association with congestive splenomegaly due to cirrhosis of the liver.
It is suggested that the association of leg ulcer with these various diseases of the blood is related in an unknown manner to either splenomegaly or hyperfunction of the spleen (hypersplenism).
THE OCCURRENCE AND SIGNIFICANCE OF "MOTILE" ERYTHROCYTES IN HUMAN BLOOD AND MARROW IN ANEMIC STATES
Authors: RALPH PAUL H. 
Published: October 2019

Abstract
1. With the aid of supravital studies and dark-field illumination, "motile" erythrocytes (capable of self initiated amoeboid-like change in form and, in some cases, movement from place to place) may be found in the peripheral blood of cases of anemia, particularly those of the macrocytic variety.
2. These motile erythrocytes are usually hypochromic, contain mitochondria and neutral red-stainable granules, all of which are attributes of normal but immature red cells.
3. Motile erythrocytes similar to those found in the blood of human patients with anemias are found in the blood of normal rabbits and mice.
4. In the marrow of patients with macrocytic anemias, megaloblasts and megalocytes may be found which give rise to motile erythrocytes by budding.
5. Poikilocytosis, motility, paleness, content of mitochondria and neutral red stainable granules, and origin by budding from a large parent cell do not differentiate the erythrocytes of pernicious anemia from those of other anemic states.
STUDIES ON HYPOPROTEINEMIA. II. FAMILIAL IDIOPATHIC DYSPROTEINEMIA
Authors: HOMBURGER F. , PETERMANN M. L. 
Published: October 2019

Abstract
1. A new syndrome, idiopathic familial dysproteinemia, is described in 4 adult members of one generation, in 2 of their paternal uncles and in 4 members of the second generation. The syndrome is characterized by hypoproteinemia and/or abnormalities in the electrophoretic patterns of the blood plasma (dysproteinemia). These are accompanied in the adult by peripheral vascular changes (ulcers of the legs in the men, low oscillometric indices in the women) and edema. There are also malformations of the thoracic cage and of the occipital hair distribution in some of the cases.
2. The idiopathic nature of the disease was ascertained in some of the patients by study of the nutritional history, of the renal, hepatic and adrenal functions, and of the response to a high-protein diet under controlled conditions.
3. In one case detailed studies of the mechanisms of plasma protein regulation resulted in findings that indicate a disturbance in the production of certain protein components. The disappearance rate of injected albumin and the rate of replacement of acutely withdrawn plasma protein were normal.
4. The clinical and physio-pathologic significance of this syndrome and the possible role of genetic factors are discussed.
Congenital neutropenia: neutrophil proliferation with abnormal maturation
Authors: Parmley RT , Ogawa M , Darby CP Jr , Spicer SS 
Published: October 2019

Abstract
A child with congenital neutropenia was studied using bone marrow culture and ultrastructural and cytochemical techniques. The patient's marrow cells formed a large number of granulocytic colonies of normal size in culture, and her peripheral blood leukocytes produced adequate colony-stimulating factor. No serum inhibitors were identified. The patient's promyelocytes from direct marrow and culture appeared normal in ultrastructure, and primary granules, contained peroxidase and acid phosphatase activity. Myelocytes and rare segmented neutrophils from direct marrow specimens demonstrated atypical notched nuclei, myelin figures in Golgi lamellae and primary (azurophilic) granules, and no identifiable secondary (specific) granules. These data indicate an intrinsic neutrophil defect which allows normal proliferation of precursor cells, but results in abnormal granulogenesis and apparent inability to form secondary granules.
Bovine factor v: a calcium-containing metalloprotein
Authors: Greenquist AC , Colman RW 
Published: October 2019

Abstract
Although coagulation factor Xa requires Ca2+ for binding to phospholipid, factor V, the other protein component in the prothrombinase enzyme complex, binds tightly to phospholipid in the absence of Ca2+. To explore the possibility that calcium might be present in the fact V molecule, the effect of several chelators, including oxalate, citrate, pyrophosphate, and EDTA, on factor V activity has been studied. A time- and concentration-dependent inhibition of factor V which reflects the respective association constants for calcium of each chelator is observed. The inhibition can be prevented by the prior addition of calcium and manganese but not magnesium. Reversal of the activity loss can be accomplished at high protein concentrations by the addition of calcium, the removal of the chelator by gel filtration, or an increase in temperature. Factor V contains 1 g atom of calcium per 300,000 daltons which is not removed by incubation with EDTA under nondenaturing conditions. Thus, the inhibition by EDTA is due to binding to calcium associated with factor V. In 8 M urea, EDTA can remove over 80% of the calcium, demonstrating the importance of the native structure in maintaining the calcium binding site. Prior binding of phospholipids to factor V prevents inhibition by EDTA. The results suggest that phospholipids complex at the calcium site on the factor V metallopretein.
High expression of transforming growth factor-beta long cell cycle times and a unique clustering of S-phase cells in patients with acute promyelocytic leukemia [see comments]
Authors: Raza A , Yousuf N , Abbas A , Umerani A , Mehdi A , Bokhari SA , Sheikh Y , Qadir K , Freeman J , Masterson M 
Published: October 2019

Abstract
Expression of transforming growth factor-beta (TGF-beta), which inhibits the proliferation of hematopoietic progenitors, was investigated simultaneously with cell cycle characteristics in 63 bone marrow biopsies from 23 cases with acute promyelocytic leukemia (APL). Bromodeoxyuridine (BrdU) was administered to every patient (17 newly diagnosed) for determination of the labeling index (LI) and the durations of S-phase (Ts) and the cell cycle (Tc) of leukemic promyelocytes. APL cases had lower LI both in the bone marrow aspirate (6.1% v 11.4%, P = .008) and biopsy (21.1% v 28.0%, P = .001) and longer Tc (93.6 hours v 56.0 hours, P = .002) when compared with other French-American-British subtypes. TGF-beta expression (detected by a monoclonal anti-TGF-beta 2/beta 3 antibody) was dramatically high, especially in interstitial areas of the biopsies. S-phase cells were found as geographically restricted islands of proliferation (GRIPs) in 20 of 22 cases. Weekly biopsies showed an increment in TGF-beta on day 7 of therapy in 13 of 17 cases, while in vivo differentiation was noted in 9 of 15. We conclude that the presence of high TGF-beta expression may explain the biologic basis for the slowly cycling nature of leukemic promyelocytes in APL as well as the unique clustering of S- phase cells observed in GRIPs.