bluuebunny/crossref_metadata_2025_split huggingface dataset:
Plasmapheresis in the Treatment of Renal Failure Associated with Multiple Myeloma.
Authors: Kumar Ambuj , Djulbegovic Benjamin , Soares Heloisa P. 
Published: October 2019

Abstract
Background: Renal failure is a common and severe complication of multiple myeloma. About 20% of the population with myeloma present with renal insufficiency and is associated with increased risk of early mortality. Plasmapheresis has been proposed as a method to reduce rate of permanent renal failure by removing the light chains from the plasma in myeloma patients. However, evidence related to the addition of plasmapheresis to chemotherapy for renal recovery in myeloma patients in existing literature is conflicting. The objective of this study is to present the totality of evidence by conducting a systematic review to assess the efficacy of plasmapheresis in preventing irreversible renal failure in patients with multiple myeloma.
Methods: A systematic and comprehensive search of the literature was performed using MEDLINE, and Cochrane library databases from 1966–2006 for all phase III randomized controlled trials comparing the use of chemotherapy with forced diuresis plus plasmapheresis versus chemotherapy plus forced diuresis in myeloma patients. Additionally, we searched ASCO and ASH meeting abstracts for the last two years. Data were extracted and pooled on overall survival and renal outcome (discontinuation of dialysis, improvement in renal function/creatinine clearence) as per the methods recommended by the Cochrane Collaboration.
Results: Altogether we identified 25 trials out of which 3 studies met the inclusion criteria. Overall, the methodological quality of the trials was good and there was no statistically significant heterogeneity between trials [outcome: overall survival (p=0.2), renal outcome (p=0.6)]. Overall survival and renal outcome were similar for the plasma exchange group and the control. Hazard ratio for the outcome overall survival was 0.76 (95% CI 0.47, 1.25, p=0.3) and for the requirement of dialysis was 0.71 (95%CI 0.35, 1.42, p=0.3) indicating no benefit for plasmapheresis in the treatment of renal failure associated with myeloma (see figure 1).
Conclusion: In multiple myeloma patients with acute renal failure there is no conclusive evidence that plasmapheresis has any effect on overall survival or on the requirement of dialysis.
Plasma exchange in renal failure for myeloma patients Plasma exchange in renal failure for myeloma patients
Role of SH2 Domain in JAK2-V617F Mediated Transformation.
Authors: Gorantla Sivahari P. , Dechow Tobias , Peschel Christian , Duyster Justus 
Published: October 2019

Abstract
A point mutation in JAK2 (V617F) has been described recently in patients with myeloproliferative diseases like polycythemia vera (PV), essential thrombocythemia (ET) and chronic idiopathic myelofibrosis (IMF). This V617F point mutation in JAK2 has been shown to activate several downstream pathways including STAT5 and ERK. This mutation also renders haematopoietic progenitors cytokine-independent. The role of the V617F mutation in oncogenesis is not fully understood. In this study we aim to dissect the role of the SH2 domain in JAK2-V617F mediated transformation.
Stable Ba/F3 cell lines expressing JAK2-wild type (wt), JAK2-V617F, JAK2-R439K (SH2 domain mutation) and JAK2-V617F/R439K mutants were generated. Cell proliferation assays showed that JAK2-V617F transforms Ba/F3 cells and renders them IL3 independent, while wild type JAK2 and JAK2-R439K could not. Surprisingly, JAK2-V617F/R439K was not able to induce a transformed phenotype in Ba/F3 cells. Imunoblotting revealed strong activation of JAK2, STAT5 and ERK in cells expressing JAK2-V617F, whereas no such activation could be found in JAK2-wt, JAK2-R439K and in JAK2-V617F/R439K expressing cells. Thus the SH2 domain in JAK2-V617F seems to play a crucial role in the transformation of Ba/F3 cells containing a heterodimeric (IL-3) cytokine receptor.
It has been demonstrated that JAK2-V617F induces cellular transformation more efficiently in cells expressing a homodimeric cytokine receptor such as the erythropoetin receptor. We therefore established Ba/F3 cells overexpressing EpoR together with JAK2-wt, JAK2-V617F, JAK2-R439K and JAK2-V617F/R439K. In contrast to parental Ba/F3 cells, EpoR expressing Ba/F3 cells could be transformed by both JAK2-V617F as well as JAK2-V617F/R439K. Both the single and double mutant Ba/F3 cells showed strong activation of STAT5 and ERK. This suggests that an intact SH2 domain is not required for homodimeric cytokine receptor expressing cells.
These results show that transformation by JAK2-V617F requires an intact SH2 domain only in cells expressing a heterodimeric cytokine receptor. In contrast, cells containing a homodimeric cytokine receptor are able to induce transformation in the presence of JAK2-V617F with an additional SH2 mutation. Further progress in understanding the role of the SH2 domain in JAK2-V617F mediated transformation may help in delineating downstream signalling with therapeutic implications.
Activating Shp2 Mutations Cause Progressive Myeloproliferative Disorder in Mice.
Authors: Mohi M. Golam , Chan Gordon , Araki Toshiyuki , Yang Wentien , Pao Lily , Kalaitzidis Demetrios , Neel Benjamin G. 
Published: October 2019

Abstract
Mutations in the SH2-containing tyrosine phosphatase, Shp2 (PTPN11) have been found in human diseases. Germ-line Shp2 mutations cause ~50% of Noonan Syndrome (NS), which is associated with an increased risk of juvenile myelomonocytic leukemia (JMML). Somatic Shp2 mutations are found in ~35% of sporadic JMML, most other cases have either activating Ras mutations or homozygous Nf1 deficiency. Shp2 mutations are also found at lower incidence in acute myeloid leukemia (AML), myelodysplastic syndrome (MDS) and B-cell acute lymphoblastic leukemia (B-ALL). NS and leukemia-associated Shp2 mutations often affect the same residue, but result in different substitution. We previously showed that leukemia-associated mutants E76K or D61Y, but not wild type (WT) Shp2 could transform bone marrow (BM) or fetal liver cells and give rise to cytokine-independent myeloid colony formation. Transplantation of E76K- or D61Y- (but not parental virus or WT Shp2) transduced BM evoked invasive MPD in most recipients and majority of them died at 6–7 months. We have generated knock-in mice for the Shp2 mutants D61G and D61Y to better understand the effects of these mutations under endogenous promoter control. Whereas D61G is associated with both NS and leukemia, D61Y is only found in leukemia and catalytically more active than D61G in basal state. D61G/+ mice exhibited all major features of NS and a mild, well-tolerated MPD for ~1 year and then progressed to fatal MPD. Constitutive expression of the D61Y heterozygous allele was embryonic lethal. However, when an inducible allele of D61Y (inD61Y) was crossed to Mx-Cre mice and expressed upon pI-pC induction, these mice rapidly developed MPD and ~50% mice died within a period of 4–7 months. Immunophenotypic analysis of bone marrow and spleen of these mice showed marked expansion of Gr-1/Mac-1 positive population indicating that these mice died of infiltrating MPD. Bone marrow macrophages from D61G/+ and inD61Y mice showed increased GM-CSF-evoked proliferation and increased overall tyrosine phosphorylation. Likewise GM-CSF-evoked activation of Erk, Akt, Jak2 and Stat5 were enhanced, as was Ras loading suggesting that leukemogenic Shp2 causes MPD by enhancing both Ras/Erk and Jak2/Stat5 pathways. The results of our retroviral and knock-in models suggest that leukemia-associated Shp2 mutant is sufficient to initiate myeloid leukemogenesis in mice.
Tlr4 Is Monoallelically Expressed in Human BFU-E and CFU-E and May Have Application as a Clonality Marker in Hematological Disorders Due to Somatic Mutations in Both Males and Females.
Authors: Chen George L. , Liu Enli , Swierczek Sabina , Prchal Josef T. 
Published: October 2019

Abstract
Tlr4 is monoallelically expressed in a tissue specific manner in mice (Peireira JP, Girard R, Chaby R, et al. Nat Immunol 4(5): 464–70, 2003). We characterized Tlr4 allelic expression in human BFU-E, CFU-E, and reticulocytes. The alleles were distinguished by a single nucleotide polymorphism (SNP) that resulted in creation of a new Bcc1 restriction enzyme cutting site. Individuals were genotyped for the SNP and only individuals heterozygous for the SNP were used in experiments. Polycythemia vera patients were diagnosed based on clinical presentation, the presence of the JAK2 V617F mutation, and clonality by X chromosome inactivation methods when available. Individual human BFU-E and CFU-E were grown from whole blood mononuclear cell fractions on methylcellulose media. Allelic expression of Tlr4 was assayed after reverse transcription of mRNA, nested PCR amplification of cDNA, and Bcc1 digestion of PCR product.
Normal human BFU-E and CFU-E colonies demonstrated monoallelic expression of both alleles as well as biallelic expression (n=1). Polycythemia vera patient BFU-E and CFU-E colonies demonstrated a similar pattern (n=2). Tlr4 allelic expression in reticulocyte RNA fractions was assayed as outlined above and by quantitative reverse transcription PCR methods. Normal human reticulocyte RNA fractions isolated from whole blood demonstrated biallelic expression (n=4). 3 of 5 polycythemia vera patient reticulocyte RNA fractions isolated from whole blood demonstrated skewed biallelic expression, and 2 of 5 polycythemia vera patients demonstrated monoallelic expression of either allele.
Tlr4 is located on chromosome 9 in humans. The finding of monoallelic expression in the reticulocyte RNA of 2 of 5 PV patients suggests thatTlr4 may be used as a phenotypic clonality marker in males and females, anda subset of polycythemia vera patients can be identified within the diagnostic population defined by clinical history and the JAK2 V617F mutation.If the assumption that clonal proliferation follows somatic mutation is true, then at least 2 mutations are implied in the pathogenesis of polycythemia vera: one by the original demonstration of clonality by X-chromosome inactivation methods, and the other by the demonstration of acquired monoallelic Tlr4 expression in the BFU-E/CFU-E to reticulocyte transition. These data are now followed by a prospective study of Tlr4 expression in a large number of female and male patients to be correlated with quantitative JAK2 V617F measurements.
SCF and IL-5 Synergize with FIP1L1/PDGFRα To Induce Mastocytosis in a Chronic Eosinophilic Leukemia Murine Model.
Authors: Yamada Yoshiyuki , Rothenberg Marc E. , Lee Andrew W. , Al-Moamen Nabeel J.H. , Williams David A. , Cancelas Jose A. 
Published: October 2019

Abstract
FIP1L1/PDGFRα (F/P) fusion protein has been identified as a cause of chronic eosinophilic leukemia (CEL)/hypereosinophilic syndrome (HES). Our group has previously described a murine CEL/HES model based on the expression of both F/P and T-cell-dependent overexpression of IL-5 (Yamada Y et al., Blood 2006), which develops a full hypereosinophilic syndrome with eosinophil infiltration into multiple organs similar to that observed in patients with CEL/HES (CEL-like mouse). Patients with F/P+ HES/CEL also display mastocytosis. Since mast cell (MC) development largely depends on c-kit signaling induced by its ligand SCF, we aimed to determine if SCF collaborates with the F/P fusion protein and IL-5 to enhance MC development. The CEL-like mice showed higher levels of MC infiltration in small intestine compared to transplanted mice with IL-5 transgenic HSC/P (control, Table 1). Interestingly, the intestinal MC infiltration of CEL-like mice was primarily associated with MC residing in the lamina propria and intraepithelial locations associated with villi; whereas MC in control mice were primarily in the crypt areas (47-fold higher mast cell levels in villi compared to those of control mice). In addition to the small intestine, skin MC infiltration was also significantly increased in CEL-like mice (Table 1). Notably, F/P+ BM hematopoietic stem cell/progenitor (HSC/P) showed proliferation and MC differentiation in vitro in the absence of cytokines (2.3-fold cell expansion, 54% c-kit+/FcεRIα+ cells) while empty vector (EGFP alone)-transduced (control) HSC/P did not survive in this culture condition. Such an expansion became even higher (36-fold expansion, 90% c-kit+/FcεRIα+ cells and 2,000-fold higher than control HSC/P) in the presence of low-concentration of SCF (10 ng/ml) for 3 weeks in culture. In contrast, culture with no cytokines or low-dose of SCF did not induce any MC development from control HSC/P. Unlike low-concentration of SCF, IL-3 (100 ng/ml), which induces a rapid and significant MC expansion in control cells, impaired MC development of F/P+ HSC/P (129-fold lower expansion at 3 weeks of culture), suggesting that MC proliferation induced by F/P expression may trigger different signaling pathways than during normal MC differentiation induced by IL-3. In summary, the F/P fusion protein induces murine mastocytosis via SCF/c-kit signaling, which is synergistically enhanced by IL-5 overexpression.
Table 1. Tissue Mast Cell Infiltration (Mast Cells/mm2) Control (IL-5 overexpressing) mice CEL-like mice *p<0.05 compared to control; n=4, per group Small Intestine 144 ± 70.2 964 ± 316.5* Skin 129 ± 33.2 208 ± 42.8*
Routine Automated Processing of Cord Blood Units Using the Sepax Cell Processing System.
Authors: Karandish Safa , Berrios Nery , Kiran Sufira , Ayuste Charisse , Hamblin Toby , Ceriani Alessio , Shpall Elizabeth J. , McMannis John D. 
Published: October 2019

Abstract
We recently incorporated the use of the automated Sepax Cell Processing System (Biosafe SA, Switzerland) for red cell and volume reduction of cord blood units (CBU) before cryopreservation. Now that we have been routinely using this new technique in the laboratory for about six months, we decided to compare the results of this method with our standard manual processing method (Rubinstein, et al, PNAS1995,; 92: 10119–22).
For both methods, hespan is added to the cells at final concentration of 20% (v/v). With the Sepax system, after addition of hespan, the cell bag is connected to the Sepax tubing set with the final freeze bag pre-attached to the set. After completion of the automated procedure, buffy coat is collected in the final freeze bag. Cryoprotectant solution is then added directly to the freeze bag. In the manual method, buffy coat and white cell rich plasma layer is collected after first centrifugation step and the white cells are separated from the plasma after the second centrifugation step. Cryoprotectant solution is then added to the cells before transfer to the final freeze bag for cryopreservation.
The following are summary of results for each method:
Table 1 Manual (n=1160) Sepax (n=311) Pre-Processing volume (ml) 107±30 114±28 Pre-Processing TNC (×10e6) 1196±577 1315±519 TNC Recovery (%) 80±8 83±8 TNC Viability (%) (7-AAD) 97±3 98±3 Total CD34 (×10e6) 4.3±4 4.9±3.6 Total DFU (×10e6) 70±0.9 61.5±20 Post Processing RBC Volume (ml) 9±2 7.3±2 Total Processing Time (including Setup) 60 minutes 30 minutes
It is important to note that there was not a significant difference in TNC Recovery over a wide range of Pre-Processing Volume (66–206ml) or Pre-Processing TNC (440 – 3559×106).
Since the Sepax device is an automated procedure, issues could arise (i.e. short term loss of electrical power) that would require us to reprocess the CBU before cryopreservation. The Sepax system allows for recovery and reprocessing of the cell using the ‘Purge mode’. We used 5 CBU units to evaluate TNC recovery and viability after purging the cells once and reprocessing.
Table 2 TNC Recovery (%) TNC Viability (%) First Buffy Coat 85±5 98±0.9 Post Purge and reprocessing 76±5 98±0.9
Although the TNC recovery was lower after the second procedure, it was still within acceptable limits and the viability of the cells had not changed. These data demonstrates that both methods are equivalent with respect to cell recovery. However, the Sepax System substantially reduces processing time and hands-on operator intervention. Additionally system provides, closed-system processing, bar code reading capability and run data print-out suitable for GMP manufacturing settings.
Context-Specific Roles for Erythroid Krüppel-Like Factor (EKLF) in Co-Ordinate High Level Expression of the Murine α- and β-Globin Genes.
Authors: Jansen Valerie M. , Ramachandran Shaji , Desgardin Aurelie , He Jin , Parekh Vishwas , Jane Stephen M. , Cunningham John M. 
Published: October 2019

Abstract
Binding of EKLF to the proximal promoter CACC motif is essential for high-level tissue-specific β-globin gene expression. More recent studies have demonstrated that EKLF regulates expression of other erythroid-specific genes, suggesting a broad role for EKLF in co-ordinating gene transcription in differentiating erythroblasts. Given these observations, we hypothesized that EKLF may play a role in synchronizing α- and β-globin gene expression. Supporting this model, studies of fetal erythroblasts derived from EKLF-null embryos revealed a 3-fold reduction in murine α-globin gene expression in fetal erythroblasts when compared to wild type littermate controls. A similar reduction in primary α-globin RNA transcripts was observed in these studies. To further examine the molecular consequences of EKLF function at the α- and β-globin genes in vivo, we utilized an erythroid cell line derived from EKLF null fetal liver cells. We have demonstrated previously that introduction into these cells of the wildtype EKLF cDNA, fused in frame with a mutant estrogen response element results in tamoxifen-dependent rescue of β-globin gene expression. Consistent with our observations in primary erythroblasts, α-globin gene expression is present in the absence of functional EKLF. However, with tamoxifen induction, we observed a 3–5 fold increase in α-globin gene transcription. Interestingly, the kinetics of the changes in transcription of the α- and β-gene transcripts were similar. Enhancement in α-gene transcription was associated with EKLF binding at the α- and β-globin promoters as determined by a quantitative chromatin immunoprecipitation (ChIP) assay. Interestingly, maximal EKLF binding and α-gene transcription was observed within 2 hours of tamoxifen induction. We hypothesized that the role of EKLF may differ function at the promoters, given that a basal level of α-globin gene expression occurs in absence of EKLF binding. Supporting this hypothesis, we observed sequential recruitment of p45NF-E2, RNA polymerase II (Pol II) and the co-activator CBP to the β-promoter with tamoxifen induction. No change in GATA-1 binding was observed. In contrast, p45NF-E2 does not bind to the α-promoter and the kinetics of GATA-1 and PolII association is unchanged after tamoxifen induction. Taken together, our results demonstrate that EKLF regulates the co-ordinate high-level transcription of the α- and β-globin genes, binding in a kinetically identical manner to the gene promoters. However, the effects of EKLF on transacting factor recruitment (and chromatin modification) differ between the promoters, consistent with the idea that EKLF acts in a context-specific manner to modulate gene transcription.
Prior Rituximab Reduces Relapse and Improves Survival Following High Dose Chemotherapy and Stem Cell Transplantation for Relapsed Composite Low and Intermediate Grade (Including Transformed) Lymphoma.
Authors: Ramadan Khaled M. , Connors Joseph M. , Al-Tourah Abdulwahab J. , Gascoyne Randy D. , Song Kevin , Barnett Michael J. , Nantel Stephen H. , Nevill Thomas J. , Shepherd John D. , Sutherland Heather J. , Lavoie Julye , Forrest Donna L. , Hogge Donna E. , Voss Nicholas J. , Brinkman Ryan , Smith Clayton A. , Toze Cynthia L. 
Published: October 2019

Abstract
The effect of prior exposure to rituximab (ritux) on relapse (REL) and survival following stem cell transplantation (SCT) in patients (pts) with REL composite low and intermediate grade non-Hodgkin lymphoma (L/I-NHL) is unknown. Fifty-four pts with REL L/I-NHL underwent high-dose chemotherapy (CT) and SCT Jan ’89 to June ’05. Follow-up is complete to April ’6 with a median of 32 mo. Ritux was added to CT regimens since 2001 and given at 375 mg/m2. Eighteen pts received ritux at initial diagnosis of NHL or after REL with salvage CT prior to SCT. Pts proceeded to high dose CT with allogeneic (allo) (n=12) or autologous (auto) SCT (n=6). This group was compared with a group of pts not receiving ritux pre-SCT (n=36)(Table 1).
Eleven pts (61%) are alive in the ritux group compared with 11 pts (31%) in the non-ritux group. The 2 and 4-y OS for pts who received ritux were 52% and 52%, vs 36% and 24% (p=.12, RR=.52) for pts who did not receive ritux. The EFS were significantly different with 2 and 4-y EFS for the ritux group 56% and 56% vs 24% and 18% (P=.038, RR=.42) for the non-ritux group (figure 1). No effect was seen on TRM. The risk of REL post SCT was significantly lower in the ritux group (p=.017)(figure 2). Two of 18 pts (11%) had NHL REL in the ritux vs 18 of 36 (50%) in the non-ritux group. The hazard rate of REL in pts who received ritux was 20% that of pts in the non-ritux group. Significance maintained in multivariate analysis (p=.016).
No impact was seen on graft-versus-host disease (GVHD). Fifty percent and 61% of pts developed acute GVHD grades 2–4 and 50% and 64% of pts developed chronic GVHD in the ritux and non-ritux groups respectively.
In conclusion, prior treatment with ritux in pts with relapsed composite L/I-NHL undergoing SCT was associated with reduced risk of REL and improved survival without associated increase in toxicity. Further analysis is underway to clarify the nature of this important finding.
Table 1: Clinicopathological characteristics of ritux and non-ritux groups Parameter$ Rituximab group, n=18 (%) Non-rituximab group, n=36 (%) $ all p values are >0.1. *at diagnosis Median age at SCT 48 y 44 y M:F 2.6:1 1.6:1 Allo-SCT 12(67) 28(78) BM involvement* 10(56) 22(61) B symptoms* 7(39) 10(28) Prior purine analogue therapy 8(44) 11(31) Residual disease prior to SCT 9(50) 14(39) TBI in conditioning 13(72) 30(83)
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